WO2020252757A1 - Pharmaceutical composition for controlling african swine fever, and extraction, injection and use thereof - Google Patents

Abstract

Disclosed is a pharmaceutical composition for controlling African swine fever, and an extraction, an injection and the use thereof. The pharmaceutical composition comprises the following raw materials in parts by weight: 10‑30 parts of ganoderma, 4‑12 parts of Drosera burmannii, 4‑12 parts of Harrisonia perforata, 30‑50 parts of Arecae semen, 4‑12 parts of Callicarpa nudiflora, 4‑12 parts of Stephania epigaea root and 4‑12 parts of nepenthes. The pharmaceutical composition has an efficient control effect on African swine fever, can be widely used to preventing the African swine fever virus, and solves a major problem of the pork industry.

Description

Medicinal composition for preventing and treating African swine fever, extract, injection and application thereof Technical field

The present invention relates to the technical field of veterinary medicine, in particular to a pharmaceutical composition for preventing and treating African swine fever, its extract, injection and application.

Background technique

African swine fever (English name: African swine fever, abbreviation: ASF) is an acute, hemorrhagic, and severe infectious disease caused by the African swine fever virus infecting domestic pigs and various wild boars (such as African wild boar, European wild boar, etc.). African swine fever does not infect humans, nor does it infect animals other than pigs, but it has a mortality rate of 100% for pigs. At present, there is no effective vaccine and treatment medicine in the world. Only culling and purification measures can be taken. African Swine Fever appeared in Kenya, south of Africa's sub-Saharan Desert in 1921. It has spread globally since the middle of the 20th century. On August 2, 2018, an African swine fever epidemic occurred in Shenbei New District, Shenyang. Since then, painful pig funerals have been staged in turn across the country. At present, epidemics have occurred in 31 provinces and cities across the country. African swine fever is the "No. 1 killer" to the global pig industry. China is a major producer and consumer of pork, with about 700 million pigs sold every year, accounting for about 50% of the world's consumption. African swine fever has seriously affected the development of the pork industry and even affected people's daily diet.

Hainan is rich in tropical plant resources. There are more than 4,000 tropical plants, accounting for 30% of the country's total. It is also one of the six hotspots of biodiversity in the world. Hainan is known as the "natural medicine library". Among these green plants, there are more than 2500 kinds of medicinal plants that are closely related to human health, more than 600 kinds of unique plants, and more than 500 kinds of plants included in the pharmacopoeia. Plant polysaccharides are compounds composed of many monosaccharides with the same or different structures through glycosidic bonds. Plant polysaccharides are widely present in plant organisms. At present, 100 plant polysaccharides have been isolated and extracted. Plant polysaccharides have a variety of biological effects, including immune regulation, anti-tumor, lowering blood sugar, lowering blood fat, anti-radiation, anti-virus, anti-aging and other functions. Plant polysaccharides promote the body's specific immunity and non-specific immunity, improve the body's disease resistance and resistance, and can also be used as an immune booster in conjunction with vaccines, which can significantly improve the protection of the vaccine and the body's disease resistance. It can also induce the production of interferon (IFN) and produce antiviral effects.

At present, there is no effective medicine to control and prevent African swine fever, so there is an urgent need for a medicine that can cure or prevent African swine fever and solve major problems in the pork industry.

Summary of the invention

In view of this, the present invention proposes a pharmaceutical composition for preventing and treating African swine fever, its extract, injection and application.

The technical scheme of the present invention is achieved as follows: a pharmaceutical composition for preventing and treating African swine fever, the pharmaceutical composition comprising the following raw materials in parts by weight: 10-30 parts of Ganoderma lucidum, 4-12 parts of Jindiluo, and beef tendon fruit 4-12 servings, 30-50 servings of betel nut, 4-12 servings of nude flower purple bead, 4-12 servings of Diwurong, 4-12 servings of Nepenthes.

Preferably, the pharmaceutical composition comprises the following raw materials in parts by weight: 18-22 parts of Ganoderma lucidum, 6-10 parts of Jindiluo, 6-10 parts of beef tendon, 36-44 parts of betel nut, and 6-10 parts of nude flower purple beads. Servings, 6-10 servings of groundwater, 6-10 servings of Nepenthes.

Preferably, the pharmaceutical composition includes the following raw materials in parts by weight: 20 parts of Ganoderma lucidum, 8 parts of Jindiluo, 8 parts of beef tendon, 40 parts of betel nut, 8 parts of nude flower purple bead, 8 parts of dianthus, and Nepenthes 8 servings.

A pharmaceutical extract for preventing and treating African swine fever comprises the pharmaceutical composition for preventing and treating African swine fever of the present invention.

Preferably, the preparation method of the pharmaceutical extract for preventing and treating African swine fever includes the following steps:

Take Ganoderma lucidum and Areca nut according to the above weight parts to prepare Ganoderma lucidum extract and Areca nut extract respectively;

According to the above weight parts, take Jindiluo, Gooseberry Fruit, Naked Flower Purple Bead, Dibumen, and Nepenthes Herbal Materials to prepare a mixed extract, including the following steps: Take Jindiluo, Gooseberry Fruit, and Naked Flower in the above weight parts Add purple bead, Dibu Rong, and Nepenthes herbal materials into the extraction container, add 8-12 times the weight of the medicinal materials, soak for 55-65min, add 0.1%-1% of the medicinal materials by weight percentage of cellulase, adjust the pH to 4～6,50-60℃ enzymatic hydrolysis for 0.5～2h, boil, extract for 1～2h, get the first extract, add 6～10 times the weight of medicinal material water, boil for the second time, extract 1～2h, get the second time Extract the two extracts, adjust the pH to 6.8～7.2 with NaOH solution, filter, concentrate the filtrate under reduced pressure to a relative density of 1.08～1.15, add 92v/v%～98v/v% ethanol solution to make the alcohol content 60%～80%, stand at 0～6℃ and refrigerate for 20～28h, centrifuge at 3000～4000r/min for 10～15min, take the precipitate, vacuum dry below 60℃ to obtain the mixed extract;

Then, the Ganoderma lucidum extract and the betel nut extract and the mixed extract are mixed to prepare a drug extract for preventing and treating African swine fever.

A medicinal injection for preventing and treating African swine fever comprises the drug extract for preventing and treating African swine fever of the present invention.

Preferably, the preparation method of the pharmaceutical injection includes the following steps: take the above-mentioned Ganoderma lucidum extract, betel nut extract, and mixed extract, mix, add water for injection to fully dissolve, add 0.8-1.2ml benzyl alcohol, 0.8- 1.2ml Tween-80, adjust the pH value to 6.8～7.2 with NaOH solution, add water for injection to make the volume; add 0.4%～0.6% injection activated carbon to remove heat source, stir well for 20～30min, and filter repeatedly after dissolution To clarify, take the filtrate, let it cool for 20-28h, filter, fill it in an ampoule, autoclave at 121°C for 10-20 minutes, and test it to obtain a drug injection. Each 1ml injection contains 1.8 equivalent of original medicinal materials. ~2.2g.

Application of the pharmaceutical composition for preventing and treating African swine fever according to any one of the present invention in the preparation of a medicine for preventing or/and curing and preventing African swine fever.

Application of the drug extract for preventing and treating African swine fever according to any one of the present invention in the preparation of drugs for preventing or/and treating African swine fever.

Application of the drug injection for preventing and treating African swine fever according to any one of the present invention in the preparation of drugs for preventing or/and treating African swine fever.

Introduction to the medicinal properties, functions or ingredients of each medicinal material of the present invention:

Ganoderma: It is the fruiting body of the porous castor family fungus Ganoderma lucidum. Ganoderma lucidum, a native medicinal material of Hainan, is rich in Ganoderma lucidum polysaccharides.

Jindiluo: It is an insectivorous plant widely distributed in tropical and subtropical areas of Asia, Africa and Oceania. It is mostly found in coastal areas or islands. The Grass Compendium records Jindiluo: mountain miasma poisoning and poisoning. , Yigenyanshengjiufu a money dagger, that is the solution (Shizhen).

Gooseberry: It is the root of Gooseberry, a plant of the bitter wood family, distributed in Hainan and Guangdong. It has the effect of clearing away heat and preventing malaria.

Betel nut: It is the seed of the palm plant Areca catechu L., a precious tropical plant in Hainan. It is located at the top of my country's five major southern medicines and contains alkaloids and polysaccharides.

Nude flower purple bead: It belongs to the Verbena family, from shrubs to small trees. It is distributed in Hainan, Guangdong and other places in my country. It has the functions of hemostasis, invigorating blood, hemostasis, removing heat, and detoxification. "Materia Medica Supplements" records: detoxification Ulcer, throat numbness, swelling, dysplasia, snake worm sting, rabies poison.

The ground is not tolerant of cold and bitter in taste, has the functions of clearing away heat, detoxifying, cutting malaria, and analgesic. It is an authentic medicinal material in Hainan.

Nepenthes: It is a tropical carnivorous plant. Its origin is mainly in the tropical regions of the Old World. It is distributed in Hainan, Guangdong and Guangxi in my country. It has the functions of clearing lungs, moisturizing dryness, running water, and detoxifying.

Compared with the prior art, the beneficial effects of the present invention are:

(1) The present invention aims at African swine fever originating in tropical regions. The selected medicinal materials are all produced in the tropical or subtropical regions of Hainan. The main components of the pharmaceutical composition of the present invention are natural polysaccharide complexes in tropical plants and South China Sea organisms, which can be directly Inhibit the virus from attaching to cells, entering cells and virus replication process, in addition, through indirect action, that is, by enhancing the body's antiviral immunity to exert its antiviral effect. The pharmaceutical composition of the present invention, as an IFN inducer, can enhance the functions of macrophages and T cells, increase the number of E-ring forming cells, induce cytokines, and promote the production of interleukin 2 (IL-2) to produce in animals Endogenous IFN to achieve antiviral purpose.

(2) The pharmaceutical composition of the present invention has a preventive effect on various viral infections such as African swine fever virus, has high efficiency and low toxicity, and at the same time enhances the body's antioxidant level, inhibits fat oxidation in muscles, reduces cell membrane damage, improves flesh color, and reduces dripping. Loss etc.

(3) The components of the pharmaceutical composition of the present invention have a synergistic effect, jointly inhibit the African swine fever virus, and have a significant effect on the treatment of African swine fever, with a cure rate of over 80.0% and up to 90.0%.

(4) The pharmaceutical composition of the present invention can cure or prevent African swine fever, has low cost, can be widely used, and can solve major problems in the pork industry.

Detailed ways

In order to better understand the technical content of the present invention, specific embodiments are provided below to further illustrate the present invention.

The experimental methods used in the embodiments of the present invention are conventional methods unless otherwise specified.

The materials and reagents used in the embodiments of the present invention can be obtained from commercial sources unless otherwise specified.

Example 1

A pharmaceutical composition for preventing and treating African swine fever, which comprises the following raw materials in parts by weight: 10 parts of Ganoderma lucidum, 4 parts of Jindiluo, 12 parts of beef tendon, 50 parts of betel nut, 4 parts of nude flower purple beads, Ground can not accommodate 4 parts, 12 parts of Nepenthes. The above-mentioned raw materials are prepared according to conventional methods (for example, direct water extraction or alcohol extraction), and the extracts are prepared into injections according to conventional methods. Each 1ml injection contains 2.0g equivalent to the original medicinal materials.

Example 2

A pharmaceutical composition for preventing and treating African swine fever, the pharmaceutical composition comprising the following raw materials in parts by weight: 30 parts of Ganoderma lucidum, 12 parts of Jindiluo, 4 parts of beef tendon, 30 parts of betel nut, 12 parts of nude flower purple beads, There are 12 parts of ground can not, 4 parts of Nepenthes. The above-mentioned raw materials are prepared according to conventional methods (for example, direct water extraction or alcohol extraction), and the extracts are prepared into injections according to conventional methods. Each 1ml injection contains 2.0g equivalent to the original medicinal materials.

Example 3

A pharmaceutical composition for preventing and treating African swine fever, the pharmaceutical composition comprising the following raw materials in parts by weight: 18 parts of Ganoderma lucidum, 6 parts of Jindiluo, 10 parts of beef tendon, 44 parts of betel nut, 10 parts of nude flower purple beads, There are 6 parts of ground not tolerance and 6 parts of Nepenthes. The above-mentioned raw materials are prepared according to conventional methods (for example, direct water extraction or alcohol extraction) to prepare extracts, and the extracts are prepared into injections according to conventional methods. Each 1ml injection is equivalent to 2.0g of original medicinal materials.

Example 4

A pharmaceutical composition for preventing and treating African swine fever, the pharmaceutical composition comprising the following raw materials in parts by weight: 22 parts of Ganoderma lucidum, 10 parts of Jindiluo, 6 parts of beef tendon, 36 parts of betel nut, 6 parts of nude flower purple beads, 10 parts of ground can not accommodate 10 parts of Nepenthes. The above-mentioned raw materials are prepared according to conventional methods (for example, direct water extraction or alcohol extraction), and the extracts are prepared into injections according to conventional methods. Each 1ml injection contains 2.0g equivalent to the original medicinal materials.

Example 5

A pharmaceutical composition for preventing and treating African swine fever, the pharmaceutical composition comprising the following raw materials in parts by weight: 20 parts of Ganoderma lucidum, 8 parts of Jindiluo, 8 parts of beef tendon, 40 parts of betel nut, 8 parts of nude flower purple beads, Ground can not accommodate 8 parts, 8 parts of Nepenthes. The above-mentioned raw materials are prepared according to conventional methods (for example, direct water extraction or alcohol extraction), and the extracts are prepared into injections according to conventional methods. Each 1ml injection contains 2.0g equivalent to the original medicinal materials.

Example 6

The formula of the pharmaceutical composition of this example is the same as that of Example 5. The difference from Example 5 is that each raw material is extracted by the following method:

(1) Preparation of Ganoderma lucidum extract: take the prescription amount of dried Ganoderma lucidum fruiting body to break the wall and pulverize, add water 35 times the weight of Ganoderma lucidum, extract at 55°C ultrasonically for 30 minutes, add cellulase with 2.0% Ganoderma lucidum weight percentage, and dilute hydrochloric acid Adjust the pH to 6.4, control the enzymatic hydrolysis temperature to 65°C, perform enzymatic hydrolysis for 68 minutes, multi-stage filtration, concentration under reduced pressure, and vacuum drying below 60°C to obtain Ganoderma lucidum extract;

(2) Preparation of betel nut extract: take the prescription amount of dried betel nut and crush it, add 8 times the weight of betel nut, soak for 60 minutes, assist with ultrasonic 200W, extract 60 minutes below 60℃, add 0.2% betel nut special enzyme for betel nut to enzymolysis , Adjust the pH to 5.5, enzymatically digest at 55℃ for 2h, filter, centrifuge at 3500r/min for 12min, take the supernatant, filter in multiple stages, concentrate the filtrate to a relative density of 1.11, add 95v/v% ethanol to make the alcohol content 50 %, stand at 3°C for 24h, centrifuge at 3500r/min for 12min, take the supernatant, concentrate to a relative density of 1.11, and vacuum dry below 60°C to obtain betel nut extract;

(3) Preparation of mixed extracts: according to the above weight parts, take Jindiluo, gooseberry fruit, nude flower purple beads, dianthus, and pitcher herbal materials into the extraction container, add 10 times the weight of the medicinal materials, infiltrate for 60 minutes, and add Cellulase of 0.5% by weight of medicinal materials, adjust pH to 5 with dilute hydrochloric acid, enzymatically hydrolyze at 55°C for 1h, boil, and extract for 1.5h to obtain the first extract, add 8 times the weight of medicinal material with water, boil twice, and extract 1.5h, get the second extract, combine the two extracts, adjust the pH to 7.0 with NaOH solution, filter, concentrate the filtrate under reduced pressure to a relative density of 1.11, add 95v/v% ethanol solution to make the alcohol content 70 %, stand at 3℃ for 24h, centrifuge at 3500r/min for 12min, take the precipitate, vacuum dry below 60℃, to obtain the mixed extract;

(4) Take the Ganoderma lucidum extract, the betel nut extract and the above-mentioned mixed extract and mix them to prepare an injection according to the conventional method, and each 1ml injection contains 2.0g equivalent to the original medicinal material.

Example 7

The formula of the pharmaceutical composition of this example is the same as that of Example 6, and the difference from Example 6 is that the Ganoderma lucidum extract, betel nut extract and mixed extract prepared by the above method are taken, mixed, and dissolved in water for injection, and each 150ml is added 1.0ml benzyl alcohol, 1.0ml Tween-80, adjust the pH to 7.0 with NaOH solution, add water for injection to make the volume; add 0.5% mass fraction of injection activated carbon (that is, 0.5g activated carbon dissolved in 100ml of sterile water) Heat source, stir well for 25min, after dissolving, filter repeatedly until clear, take the filtrate, cool it for 24h, filter (0.22um nylon microporous membrane), fill it in an ampoule, autoclave at 121℃ for 15 minutes, check, Obtained injection, each 1ml injection contains 2.0g equivalent to the original medicinal materials.

In order to verify the beneficial effects of the present invention, the following tests were carried out. The test methods and results are as follows:

(1) Experimental animals

A farm has a batch of live pigs, sows of 1-5 gestational age and Duroc boars, weighing 50-200kg, some of them have high fever, breathing difficulties, cough, runny nose and eye secretions, skin cyanosis, depression, anorexia, The sow has symptoms such as abortion. Treatment with a variety of antibiotics and antipyretics was ineffective and he died on the 8th day. Symptoms and necropsy confirmed infection with African swine fever.

(2) Clinical trials

Under the guidance of veterinary medicine experts, 420 sick pigs of this batch were randomly selected as the administration group. They were divided into 7 groups, each with 60 pigs. The injections prepared in Examples 1-7 of the present invention were used respectively. In the test, pigs were injected intramuscularly (neck side behind the base of the ear), 20ml/time (40ml/time for a body weight of more than 100kg), used 3 times in a row, the interval between the two intramuscular injections was 2 days, and the first intramuscular injection was doubled. Twenty sick pigs from this batch were randomly selected and fed as a control group.

(3) Treatment results

The record of the treatment effect after administration is as follows:

The statistical results are as follows:

To	Total (only)	Cure (only)	Death (only)	Cure rate	mortality rate
Example 1	60	48	12	80.0%	20.0%
Example 2	60	48	12	80.0%	20.0%
Example 3	60	50	10	83.3%	16.7%
Example 4	60	49	11	81.7%	18.3%
Example 5	60	52	8	86.7%	13.3%
Example 6	60	53	7	88.3%	11.7%
Example 7	60	54	6	90.0%	10.0%
Control group	20	0	20	0.0%	100.0%

The above-mentioned clinical test shows that the pharmaceutical composition of the present invention has a significant effect on the treatment of African swine fever, with a cure rate of over 80.0% and up to 90.0%. The pharmaceutical composition of the invention can effectively inhibit the deterioration of the disease, restore appetite, restore the damaged vitality of the body, and restore normal growth and development. It can be seen that the pharmaceutical composition of the present invention can be applied to the preparation of drugs for treating African swine fever virus.

In addition, under the supervision of relevant competent authorities, 60 healthy pigs were randomly selected, and the injection prepared in Example 7 of the present invention was used for the experiment. The pigs were injected intramuscularly (the base of the ear and the neck side), 20ml/time (with a weight of more than 100kg) 40ml/time). After one injection, the healthy pigs were placed in the African swine fever virus environment. 25 days later, 2 died and 58 remained healthy and grew normally. Clinical trials have shown that healthy pigs injected with the injection prepared in Example 7 of the present invention can prevent infection of African swine fever virus. It can be seen that the pharmaceutical composition of the present invention can also be used to prepare drugs for preventing infection of African swine fever virus.

The above descriptions are only the preferred embodiments of the present invention and are not intended to limit the present invention. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the present invention. Within the scope of protection.

Claims (10)

1. A pharmaceutical composition for preventing and treating African swine fever, characterized in that the pharmaceutical composition comprises the following raw materials in parts by weight: 10-30 parts of Ganoderma lucidum, 4-12 parts of Jindiluo, 4-12 parts of beef tendon, and betel nut 30-50 servings, 4-12 servings of nude flower purple bead, 4-12 servings of Diwurong, 4-12 servings of Nepenthes.
2. The pharmaceutical composition for preventing and treating African swine fever according to claim 1, wherein the pharmaceutical composition comprises the following raw materials in parts by weight: 18-22 parts of Ganoderma lucidum, 6-10 parts of Jindi Luo, beef tendon 6-10 parts of fruit, 36-44 parts of betel nut, 6-10 parts of nude flower purple bead, 6-10 parts of Diwurong, 6-10 parts of Nepenthes.
3. The pharmaceutical composition for preventing and treating African swine fever according to claim 2, wherein the pharmaceutical composition comprises the following raw materials in parts by weight: 20 parts of Ganoderma lucidum, 8 parts of Jindi Luo, 8 parts of beef tendon, 40 betel nut, 8 nude flower purple bead, 8 dicanthus, 8 nepenthes.
4. A pharmaceutical extract for preventing and treating African swine fever, characterized in that it comprises the pharmaceutical composition for preventing and treating African swine fever according to claim 1.
5. The medicinal extract for preventing and treating African swine fever according to claim 4, wherein the preparation method comprises the following steps:

   Take Ganoderma lucidum and Areca nut according to the above weight parts to prepare Ganoderma lucidum extract and Areca nut extract respectively;

   According to the above weight parts, take Jindiluo, Gooseberry Fruit, Naked Flower Purple Bead, Dibumen, and Nepenthes Herbal Materials to prepare a mixed extract, including the following steps: Take Jindiluo, Gooseberry Fruit, and Naked Flower in the above weight parts Add purple bead, Dibu Rong, and Nepenthes herb materials to the extraction container, add 8-12 times the weight of the medicinal materials, soak for 55-65min, add 0.1%-1% cellulase by weight of the medicinal materials, and adjust the pH to 4-6 , 50-60℃ enzymatic hydrolysis for 0.5-2h, boiling, extracting for 1-2h, get the first extract, add 6-10 times the weight of medicinal material water, boil for the second time, extract for 1-2h, get the second extract, Combine the two extracts, adjust the pH to 6.8～7.2, filter, and concentrate the filtrate under reduced pressure to a relative density of 1.08～1.15. Add ethanol solution to make the alcohol content 60%～80%, and keep it at 0～6℃ for 20～ Centrifuge for 28h, take the precipitate, and dry it in vacuum below 60°C to obtain a mixed extract;

   Then, the Ganoderma lucidum extract and the betel nut extract and the mixed extract are mixed to prepare a drug extract for preventing and treating African swine fever.
6. A pharmaceutical injection for preventing and treating African swine fever, which is characterized in that it contains the pharmaceutical extract for preventing and treating African swine fever according to claim 5.
7. The pharmaceutical injection for preventing and treating African swine fever according to claim 6, characterized in that the preparation method comprises the following steps: taking the above-mentioned Ganoderma lucidum extract, betel nut extract and mixed extract, mixing, adding water for injection to fully dissolve , Add 0.8-1.2ml benzyl alcohol, 0.8-1.2ml Tween-80 per 150ml, adjust the pH to 6.8-7.2, add water for injection to make the volume; add 0.4% to 0.6% injection activated carbon to remove heat source, fully Stir for 20-30min, after dissolution is complete, filter repeatedly until clear, take the filtrate, cool it for 20-28h, filter, fill in an ampoule, autoclave at 121°C for 10-20 minutes, test, prepare injection, 1ml contains 1.8～2.2g equivalent of original medicinal materials.
8. The use of the pharmaceutical composition for preventing and treating African swine fever according to any one of claims 1 to 3 in the preparation of a medicine for preventing or/and curing and preventing African swine fever.
9. The use of the pharmaceutical extract for preventing and treating African swine fever according to any one of claims 4 to 5 in the preparation of a medicine for preventing or/and treating African swine fever.
10. The use of the pharmaceutical injection for preventing and treating African swine fever according to any one of claims 6 to 7 in the preparation of drugs for preventing or/and treating African swine fever.