CN115887459A - Application of cepharanthine and pharmaceutical composition thereof in preparation of anti-African swine fever drugs - Google Patents
Application of cepharanthine and pharmaceutical composition thereof in preparation of anti-African swine fever drugs Download PDFInfo
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- CN115887459A CN115887459A CN202310088241.5A CN202310088241A CN115887459A CN 115887459 A CN115887459 A CN 115887459A CN 202310088241 A CN202310088241 A CN 202310088241A CN 115887459 A CN115887459 A CN 115887459A
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- swine fever
- african swine
- cepharanthine
- fever virus
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- YVPXVXANRNDGTA-WDYNHAJCSA-N cepharanthine Chemical compound C1C(C=C2)=CC=C2OC(=C2)C(OC)=CC=C2C[C@H](C2=C3)N(C)CCC2=CC(OC)=C3OC2=C(OCO3)C3=CC3=C2[C@H]1N(C)CC3 YVPXVXANRNDGTA-WDYNHAJCSA-N 0.000 title claims abstract description 82
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The invention belongs to the field of antiviral drugs, and relates to application of cepharanthine and a pharmaceutical composition thereof in preparation of an African swine fever virus resistant drug; the cepharanthine has very obvious effect on resisting African swine fever virus; experiments prove that the cepharanthine has obvious antiviral activity in cell experiments, and can have extremely strong anti-African swine fever virus (MOI = 1) infection effect at the concentration of 4 mu M. The cepharanthine has a very obvious effect of resisting African swine fever virus, is safe, has few toxic and side effects, and has low drug residue and no pollution.
Description
Technical Field
The invention belongs to the field of antiviral drugs, and particularly relates to an application of cepharanthine and a pharmaceutical composition thereof in preparation of a drug for resisting African swine fever viruses.
Background
African Swine Fever (ASF) is an acute, febrile, highly contagious and virulent infectious disease of Swine caused by African Swine Fever Virus (ASFV) infection. The African swine fever is an animal epidemic disease which is required to be legally reported by the world animal health Organization (OIE), and is classified as a disease by the national animal pathogenic microorganism list.
African swine fever virus belongs to the family African swine fever virus (Asfarviridae), the genus African swine fever virus (Asfivirus), is the only member of this family. Analysis of ASFV isolates in different regions using the B646L gene encoding VP72 protein can classify the circulating strains of ASFV into 22 major genotypes, among which the circulating strains in China are mainly genotype II.
The African swine fever virus is widely distributed in various tissues and organs and various body fluids of sick pigs, and secretion and excrement contain a large amount of virus. The virus is stable at low temperature and does not resist high temperature. Can survive for years at 4 ℃ and for months at room temperature in the presence of proteins. There was no inactivation at 60 ℃ for 20 minutes and at 56 ℃ for 30 minutes.
The African swine fever is first reported in 1921, and no effective commercial vaccine and specific medicine are developed so far. Vaccine research shows that the inactivated vaccine has almost no protective effect; vector or subunit seedlings provide only partial protection; attenuated seedlings are the most promising but have safety problems. The ASFV genome is about 170-190kbp in length and complex in structure, encodes 150-200 proteins, about 50% of the proteins are unknown in function, and a wide immune escape phenomenon exists, which causes great difficulty in vaccine development. At present, a medicine for treating the African swine fever quickly, thoroughly, efficiently and safely is not developed, and only a mode of killing the African swine fever immediately and isolating the African swine fever to prevent infection is adopted.
Cepharanthine (CEP) is a dibenzylisoquinoline alkaloid extracted from Stephania cepharantha, stephania delavayi and Stephania cepharantha of Stephania, and is called Stephania delavayi Hemsl, sifaansheng, sofoscarnine and Cepharanthine. CEP can inhibit proliferation of colorectal cancer cells and melanoma cells, induce apoptosis of lung cancer cells, reverse drug resistance of chemotherapeutic drugs, and other wide pharmacological actions. At present, no research report about the inhibition of the cepharanthine on African swine fever virus exists, and the invention aims to provide the application of the cepharanthine in the preparation of the African swine fever virus resistant medicament.
Disclosure of Invention
Object of the Invention
In the prior art, reports of using cepharanthine as an antiviral drug exist, and at present, no research report about the suppression of African swine fever virus by cepharanthine exists. The invention aims to provide a pharmaceutical composition for resisting African swine fever virus and application of cepharanthine in preparation of a drug for resisting African swine fever virus.
Technical scheme
The above object of the present invention is achieved by the following technical solutions:
in a first aspect of the invention, the use of cepharanthine in the preparation of a medicament for the treatment of African swine fever virus.
The invention also provides a pharmaceutical composition for resisting African swine fever virus, which comprises the cepharanthine and a pharmaceutically acceptable carrier.
In some preferred embodiments, the anti-african swine fever virus drug may be a tablet, a powder, a granule, a capsule, an oral liquid, an injection or a sustained release agent.
Optionally, the african swine fever virus is a genotype II african swine fever virus.
Optionally, the excipient is selected from solid excipients or liquid and semi-solid excipients.
To prepare the pharmaceutical composition of the present invention, cepharanthine may be mixed with a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers that can be used in the present compositions encompass any standard pharmaceutical carrier, such as aqueous phosphate buffered solutions, water and emulsions, such as oil/water or water/oil emulsions, aqueous dextrose, glycols and various types of wetting agents.
The pharmaceutical compositions of the present invention may additionally comprise solid excipients, liquid excipients and semisolid excipients. The solid excipient can be selected from starch, cellulose, talc, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, magnesium stearate, sodium stearate, glycerol monostearate, sodium chloride, dried skim milk, etc. The liquid and semi-solid excipients may be selected from glycerol, propylene glycol, water, ethanol, and various oils, including those of petroleum, animal, vegetable, or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil, and the like. The pharmaceutical compositions of the present invention may also include stabilizers and preservatives.
The present compositions may be administered by any method known in the art, including, but not limited to, intranasal, oral, transdermal, ocular, intraperitoneal, inhalation, intravenous, intracisternal injection or infusion, subcutaneous, implant, intravaginal, sublingual, urethral, subcutaneous, intramuscular, intravenous, rectal, sublingual, mucosal, ocular, spinal, intrathecal, intra-articular, intra-arterial, subarachnoid, bronchial and lymphatic administration. The external preparation may be in the form of gel, ointment, cream, aerosol, etc.; intranasal formulations may be delivered in spray or droplet form; transdermal formulations may be administered via transdermal patches or ion permeation therapy; inhalation formulations may be delivered using a nebulizer or similar device. The compositions may also be in the form of tablets, capsules, suspensions, solutions, emulsions, injections, ointments, gels, films, pills, granules or powders or any other suitable composition.
Compared with the prior art, the invention has the beneficial effects that:
1. the cepharanthine has very obvious effect of resisting African swine fever virus; experiments prove that the cepharanthine has obvious antiviral activity in cell experiments, and can have extremely strong anti-African swine fever virus (MOI = 1) infection effect at the concentration of 4 mu M.
2. The cepharanthine is safe and has little toxic and side effects when being used for resisting African swine fever virus; the cepharanthine is extracted from Chinese herbal medicine, is different from hormone, antibiotic, chemical synthetic medicine, etc., and has no obvious toxic and side effect on organism.
3. The cepharanthine is used for resisting African swine fever virus and has low drug residue and no pollution; cepharanthine belongs to organic molecular compound, and is easily absorbed by animal body, and has high biological metabolism rate and no pollution.
Drawings
To more clearly illustrate the technical solutions of the embodiments of the present disclosure, the drawings that are required to be used in the embodiments will be briefly described below, it should be understood that the following drawings only illustrate certain embodiments of the present disclosure and therefore should not be considered as limiting the scope, and that for those skilled in the art, other related drawings may be obtained from these drawings without inventive effort.
FIG. 1 shows the results of the CCK-8 method for detecting toxicity of cepharanthine to PAM cells. The PAM cells are treated by 0.5-20 mu M cepharanthine, and then the activity of the cells is detected by using a CCK-8 method and measuring the absorbance value of the cells at 450 nm.
FIG. 2 shows the results of enzyme-linked immunosorbent assay (ELISA) for detecting the inhibition of ASFV infection by cepharanthine on PAM cells. 10 μ M cepharanthine pre-treated PAM cells for 2 hours were infected with African swine fever virus, then cepharanthine was present all the time, and cells were fixed for ELISA 48 hours later.
FIG. 3 shows the activity of WesternBlot in the inhibition of African swine fever virus infection on PAM cells. mu.M, 0.5. Mu.M, 1. Mu.M, 2. Mu.M, 4. Mu.M cepharanthin pre-treated PAM cells for 2 hours followed by African swine fever virus infection, followed by cepharanthin persistence, and 24 hours later cells were harvested for WesternBlot determination of the content of the viral protein ASFV-p72 in the cells.
FIG. 4 shows the activity of QPCR in the inhibition of African swine fever virus infection by cepharanthine on PAM cells. P72mRNA levels of African swine fever virus were detected using QPCR after pretreatment of PAM cells for 2 hours with 1.25. Mu.M, 2.5. Mu.M and 5. Mu.M cepharanthin, after which cepharanthin was present and 48 hours later total cellular RNA was extracted and reverse transcribed to cDNA.
FIG. 5 shows WesternBlot results of the addition of cepharanthine at different stages of African Swine Fever Virus (ASFV) virus infection;
FIG. 6 shows the number of fluorescent cells in cells treated with cepharanthine (2. Mu.M and 4. Mu.M) at different concentrations after 24 h infection of PAM cells by Rinderpest virus (CHINA/2018/AnguiXCGQ).
FIG. 7 shows the number of fluorescent cells of African swine fever virus (CHINA/2018/AnhuiXCGQ) that entered the cells after treatment with different concentrations of cepharanthine (1. Mu.M, 2. Mu.M and 4. Mu.M).
Detailed Description
The following claims are presented in further detail in connection with experiments and examples, and are not to be construed as limiting the disclosure in any way, and any limited number of modifications that can be made within the scope of the claims are intended to be within the scope of the disclosure. The embodiment does not indicate specific conditions, and the method is carried out according to conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
Unless defined otherwise herein, scientific and technical terms used in connection with the present disclosure shall have the meanings that are commonly understood by one of ordinary skill in the art. Exemplary methods and materials are described below, but methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present disclosure.
Abbreviations
Cepharanthine: CEP
African swine fever virus: ASFV
Test materials
The African swine fever virus CHINA/2018/AnhuiXCGQ strain is stored in an African swine fever reference laboratory (Shandong Qingdao) of the China center for animal health and epidemiology;
PAM cells (porcine alveolar macrophages) were prepared and stored in the inventor's laboratory;
the CCK-8 detection kit is purchased from Donglian chemical technology (Shanghai) Co., ltd;
cepharanthine (Cepharanthine) for cell experiments was purchased from Selleck. Cepharanthine (Cepharanthine) for animal experiments was purchased from Shanghai-derived Phyllobiology, inc.
Example 1 enzyme-linked immunosorbent assay (ELISA) for determining the Activity of Cepharanthine on PAM cells to inhibit African Swine fever Virus infection
After PAM cell count was diluted to appropriate density with 8% Fetal Bovine Serum (FBS) RPMI-1640 medium and applied to 96-well plates at a concentration of 1.5X 105/well, placed in a 5% CO2 incubator at 37 ℃ for 5 hours after cells adhered to a monolayer, cepharanthine was diluted to 0.0625. Mu.M, 0.125. Mu.M, 0.25. Mu.M, 0.5. Mu.M, 1. Mu.M, 2. Mu.M, 4. Mu.M, 8. Mu.M and 10. Mu.M with RPMI-1640 medium. After pre-treating PAM cells for 2 hours at 37 ℃ with DMSO blank and varying concentrations of cepharanthine, inoculated with African swine fever virus CHINA/2018/AnhuiXCGQ strain (MOI = 1), placed in a 37 ℃ 5-pot CO2 incubator for 1 hour, washed three times with PBS, added with 2% Fetal Bovine Serum (FBS) in RPMI-1640 medium 1mL, and placed in a 37 ℃ 5-pot CO2 incubator in the presence of cepharanthine for 48 hours. Discard the medium supernatant, wash three times with PBS, add 3% BSA150. Mu.L and block for 2 hours. Washed once with PBS, diluted 100 μ LASFV mab was added and incubated for 2 hours at 37 ℃. PBS was washed three times, and diluted 100. Mu.L secondary antibody was added and incubated for 40 minutes. PBS was washed three times, 100. Mu.L of TMB developing solution was added to react for 10 minutes, 50. Mu.L of stop solution was added to stop the reaction, and OD450 was read at 450 nm.
As a result:
as shown in fig. 1, the OD450 value of the experimental group treated with 10 μ M cepharanthine was very close to the OD450 value of the blank control group, demonstrating that 10 μ M cepharanthine showed a very strong inhibitory effect on african swine fever virus.
As shown in FIG. 2, different concentrations of cepharanthine (2-10 μ M) showed significant inhibition of African swine fever virus. The IC50 value of the inhibitory activity of cepharanthine against African swine fever virus obtained by GraphPadPrismv8.0 software was 0.3223. Mu.M. The invention has the ideal IC50 value of micromolar grade, and fully proves that the cepharanthine has strong activity against African swine fever virus.
Wherein the ASFV virus relative content% is calculated by the following formula:
relative viral content% = (OD 450 experimental-OD 450 blank) ÷ (OD 450 negative control-OD 450 blank) × 100%.
The OD450 test group indicated the OD450 values of wells inoculated with african swine fever virus and supplemented with cepharanthine, the OD450 negative control indicated the OD450 values of wells inoculated with african swine fever virus only and not supplemented with cepharanthine, and the OD450 blank control indicated the OD450 values of blank control wells supplemented with DMSO only.
Example 2WesternBlot assay of the Activity of Cepharanthine on PAM cells to inhibit African Swine fever Virus infection
After PAM cell count, diluted to appropriate density with 8% Fetal Bovine Serum (FBS) RPMI-1640 nutrient solution and then diluted to 2X 10 6 Concentration/well to 6-well plate, put in a CO2 incubator at 37 ℃ and 5% until the cells adhere to a monolayer density (about 8-10 hours), cepharanthine was diluted with RPMI-1640 nutrient solution to: 0.25. Mu.M, 0.5. Mu.M, 1. Mu.M, 2. Mu.M and 4. Mu.M. After pre-treating PAM cells for 2 hours at 37 ℃ with DMSO control and varying concentrations of cepharanthine, the African swine fever virus CHINA/2018/AnhuiXCGQ strain (MOI = 1) was inoculated and placed at 37 ℃ for 5% CO 2 After 1 hour in the incubator, washing three times with PBS, adding 1mL of RPMI-1640 maintenance solution containing 2% Fetal Bovine Serum (FBS), and incubating at 37 ℃ for 24 hours in the presence of cepharanthine in a 5% CO2 incubator, cell samples were harvested using 2 XSDSsample buffer, metal bath boiled for 10 minutes, westernBlot assay was performed, and the results are shown in FIG. 3.
FIG. 3 shows the content of the virus protein ASFV-p72 of African Swine Fever Virus (ASFV) in cells treated with cepharanthin (0.25-4 μ M) at different concentrations, indicating that cepharanthin can significantly inhibit African swine fever virus infection and show significant dose dependence.
Example 3QPCR assay of Activity of Cepharanthine on PAM cells to inhibit African Swine fever Virus infection
After PAM cell count, diluted to appropriate density with 8% Fetal Bovine Serum (FBS) RPMI-1640 nutrient solution and then diluted to 2X 10 6 Concentration in the poresThe cells were plated in 6-well plates, placed in a CO2 incubator at 37 ℃ and 5% after adherence to monolayer density (about 8-10 hours), and then diluted with RPMI-1640 medium to: 1.25. Mu.M, 2.5. Mu.M and 5. Mu.M. After pre-treating PAM cells for 2 hours at 37 ℃ with DMSO control and varying concentrations of cepharanthine, inoculating African swine fever virus CHINA/2018/AnhuiXCGQ strain (MOI = 1), placing in a 37 ℃ 5% CO2 incubator for 1 hour, washing three times with PBS, adding 1mL of RPMI-1640 maintenance solution containing 2% Fetal Bovine Serum (FBS), incubating in a 37 ℃ 5% CO2 incubator for 24 hours in the presence of cepharanthine, collecting cell samples with Trizon, and extracting RNA. The extracted RNA is dissolved in 20 mul of ultrapure water, inverted into cDNA, and the change of the mRNA level of ASFV-p72 protein is detected by real-time fluorescence quantitative PCR using the Actin gene as an internal reference. Wherein the primer information is as follows:
Actin-F5’CTGAAGTACCCCATCGAGCACGGCA’3’；
Actin-R5’GGATAGCACAGCCTGGATAGCAACG’3’；
ASFV-p72-F5’CCCAGGRGATAAAATGACTG3’；
ASFV-p72-R5’CACTRGTTCCCTCCACCGATA3’。
20 μ LPCR reaction System containing 10 μ l AceQ
Green Master Mix, 0.4. Mu.L upstream and downstream primers (10. Mu.M), 2. Mu.L template DNA,0.4mL ROXReferenceDye1 and 6.8. Mu.L sterile distilled water. The amplification parameters were: 40 cycles of 95 ℃ 10s and 60 ℃ 30s, and after the reaction is finished, the dissolution curve is measured, and the reaction conditions are as follows: 95 ℃ for 15s, 60 ℃ for 60s and 95 ℃ for 15s. The relative quantification of the ASFV-p72 gene is carried out by a 2-delta Ct method in the experiment;
the results are shown in fig. 4, where fig. 4 shows the relative expression levels of the viral protein ASFV-p72mRNA of African Swine Fever Virus (ASFV) in cells after treatment with different concentrations of cepharanthine (1.25-5 μ M), indicating that cepharanthine is able to significantly inhibit infection by african swine fever virus and exhibits significant dose dependence.
Example 4: specific action stage of Western Blot for determining inhibition of African swine fever virus infection of cepharanthine on PAM cells
PAM cells or PBMC cells were counted, diluted to appropriate density with 8% Fetal Bovine Serum (FBS) RPMI-1640 medium, and then diluted to 3X 10 6 Concentration per well was determined by adding to a 6-well plate containing a slide, placing at 37 ℃ and 5% CO 2 The experiment was started after the cells attached to a monolayer density (about 8-10 hours) in the incubator. The result of Western Blot detection was shown in FIG. 5, which was performed by diluting cepharanthine to 4. Mu.M with RPMI-1640 nutrient solution, inoculating PAM cells with the strain dose of African swine fever virus CHINA/2018/AnhuiXCGQ (MOI = 1), treating or infecting the cells with cepharanthine in the treatment pattern shown in FIG. 5, collecting the cell sample with 2 XSDS sample buffer after each treatment stage, boiling the sample in a metal bath for 10 minutes, and performing the Western Blot detection.
FIG. 5 shows that the addition of cepharanthine (4. Mu.M) at different stages of African Swine Fever Virus (ASFV) virus infection can inhibit the content of the virus protein ASFV-p72 of African Swine Fever Virus (ASFV) in cells to different degrees, which indicates that the cepharanthine can significantly inhibit the African swine fever virus from infecting PAM cells at various stages of virus replication.
Example 5: IFA (enzyme-linked immunosorbent assay) for determining activity of cepharanthine on PAM (polyacrylamide gel) cells for inhibiting African swine fever virus infection
PAM cells or PBMC cells were counted, diluted to appropriate density with 8% Fetal Bovine Serum (FBS) RPMI-1640 medium, and then diluted to 2X 10 6 Concentration/well to 6-well plate containing slide glass, placed in a 5-percent CO2 incubator at 37 ℃ until the cells adhere to a monolayer density (about 8-10 hours), cepharanthine was diluted 2. Mu.M and 4. Mu.M with RPMI-1640 nutrient solution. After pre-treating PAM cells for 2 hours at 37 ℃ with DMSO control and with stephanicin of different concentrations, inoculating African swine fever virus CHINA/2018/AnhuiXCGQ strain (MOI = 1), placing in an incubator with 37 ℃ and 5% CO2 for 1 hour, washing three times with PBS, adding 1mL of RPMI-1640 maintenance solution containing 2% Fetal Bovine Serum (FBS), placing in an incubator with 37 ℃ and 5% CO2 in the presence of stephanicin for 24 hours, and performing indirect immunofluorescence experiment, which comprises the following specific steps: fixing PAM cells at 37 deg.C for 30 min, eluting with PBS for 15 min, blocking at 37 deg.C for 30 min, reacting p72 primary antibody at 37 deg.C for 1 hr, eluting with PBS for 15 min, reacting secondary antibody for 30 min, and eluting with PBSAfter 15 minutes, the specimen was mounted on a mounting plate and observed under a fluorescence microscope, and the results are shown in FIG. 6.
FIG. 6 shows the number of fluorescent cells after infection of PAM by African swine fever virus (CHINA/2018/AnhuiXCGQ) in cells treated with 2. Mu.M and 4. Mu.M cepharanthin, indicating that cepharanthin can significantly inhibit infection by African swine fever virus and shows obvious dose dependence.
Example 6: IFA (enzyme-linked immunosorbent assay) determination of inhibition of African swine fever virus entry of cepharanthine on PAM (polyacrylamide) cells
PAM cells or PBMC cells were counted, diluted to appropriate density with 8% Fetal Bovine Serum (FBS) RPMI-1640 medium, and then diluted to 2X 10 6 Concentration/well to 6-well plate containing slide glass, placed in a 5-degree CO2 incubator at 37 ℃ until the cells adhere to a monolayer density (about 8-10 hours), cepharanthine was diluted with RPMI-1640 nutrient solution at 1. Mu.M, 2. Mu.M and 4. Mu.M. After pre-treating the PAM cells for 2 hours at 37 ℃ with DMSO control and varying concentrations of cepharanthine, the cells were infected with the African swine fever virus CHINA/2018/AnhuiXCGQ strain (MOI = 10), placed in a 37 ℃ 5-pot CO2 incubator for 2 hours, washed once with pre-cooled PBS and three times with PBS, and subjected to an indirect immunofluorescence assay, with IFA as specified in example 5, and the results are shown in FIG. 7.
Figure 7 shows the number of fluorescent cells after admission of african swine fever virus (CHINA/2018/AnhuiXCGQ) into PAM in 1, 2 and 4 μ M cepharanthin treated cells, indicating that cepharanthin can significantly inhibit the admission of african swine fever virus and show significant dose dependence.
The operation related to the African swine fever virus infection is carried out in a biosafety third-level laboratory of China centers for animals and epidemiology.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (8)
1. Application of cepharanthine in preparing medicine for resisting African swine fever virus is provided.
2. Application of stephania japonica extract in preparing anti-African swine fever virus drugs, wherein the extract contains stephanine.
3. An application of a traditional Chinese medicine composition in preparing a medicine for resisting African swine fever virus is characterized in that the traditional Chinese medicine composition comprises cepharanthine and a pharmaceutically acceptable carrier.
4. The use according to any one of claims 1 to 3, wherein the African swine fever virus is genotype II African swine fever virus.
5. The use according to claim 3, wherein the pharmaceutically acceptable carrier is selected from a solid excipient, a liquid or a semi-solid excipient.
6. Use according to claim 3, wherein the solid excipient is selected from the group consisting of starch, cellulose, talc, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, magnesium stearate, sodium stearate, glycerol monostearate, sodium chloride, dried skim milk or a combination of any two or more of the foregoing.
7. The use according to claim 3, wherein the liquid or semi-solid is selected from the group consisting of glycerol, propylene glycol, water, ethanol, oil, and combinations of any two or more thereof, wherein the oil is of petroleum, animal, vegetable, or synthetic origin. .
8. The use of claim 3, wherein the pharmaceutical composition is in a dosage form selected from the group consisting of tablets, capsules, suspensions, solutions, emulsions, injections, ointments, gels, films, pills, granules and powders.
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| CN115770245A (en) * | 2022-12-28 | 2023-03-10 | 华南农业大学 | Application of dibenzyl isoquinoline alkaloid in preparation of drug for preventing and treating African swine fever virus |
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| CN115770245A (en) * | 2022-12-28 | 2023-03-10 | 华南农业大学 | Application of dibenzyl isoquinoline alkaloid in preparation of drug for preventing and treating African swine fever virus |
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