CN115531384B - Application of dibenzyl isoquinoline alkaloid in preparation of African swine fever virus resistant medicaments - Google Patents
Application of dibenzyl isoquinoline alkaloid in preparation of African swine fever virus resistant medicaments Download PDFInfo
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- CN115531384B CN115531384B CN202211021536.2A CN202211021536A CN115531384B CN 115531384 B CN115531384 B CN 115531384B CN 202211021536 A CN202211021536 A CN 202211021536A CN 115531384 B CN115531384 B CN 115531384B
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- Prior art keywords
- asfv
- dibenzyl
- acid
- swine fever
- african swine
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/47—Quinolines; Isoquinolines
- A61K31/472—Non-condensed isoquinolines, e.g. papaverine
- A61K31/4725—Non-condensed isoquinolines, e.g. papaverine containing further heterocyclic rings
-
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Abstract
The invention belongs to the technical field of veterinary medicines, and in particular relates to an application of dibenzyl isoquinoline alkaloids in preparation of an anti-African Swine Fever Virus (ASFV) medicine. The dibenzyl isoquinoline alkaloid comprises the following components: the liensinine, isoliensinine, neferine, dauricine, stephanine, tetrandrine, fangchinoline and berbamine can obviously inhibit the proliferation of ASFV, and the half effective inhibition concentration of ASFV is 0.31-2.34 mu M. In addition, the dibenzyl isoquinoline alkaloid has small toxic effect on the natural target cell of the ASFV, namely the pig alveolar macrophage, can have remarkable antiviral activity on the ASFV under the concentration without cytotoxicity, and has high safety; in addition, the dibenzyl isoquinoline alkaloid is derived from common traditional Chinese medicines, has wide sources and low cost, and is very suitable for developing medicines for preventing and treating african swine fever.
Description
Technical Field
The invention belongs to the technical field of veterinary medicines. More particularly relates to an application of dibenzyl isoquinoline alkaloid in preparing an African swine fever virus resistant medicament.
Background
African swine fever (African swine fever, ASF) is a very infectious viral disease caused by infection of African swine fever virus (African swine fever virus, ASFV), the disease incidence rate is high, the diseased pigs often show acute, febrile and hemorrhagic clinical symptoms, organs of the disease show serious vascular lesions, such as lymph node hemorrhage, kidney hemorrhage, disseminated intravascular coagulation, thrombocytopenia and the like, and the death rate is close to 100%. ASF is a legal notice of disease by the world animal health Organization (OIE), a type of animal epidemic in our country, and is listed in the national long-term animal epidemic prevention program (2012-2020) as a priority for the prevention and treatment of epidemic diseases. African swine fever was first discovered in African Kenney in 1921; in 1957, ASF first erupted into grape teeth outside of the continental africa; in 2007, ASFV was again transferred from continental africa into the caucasian region bordered by eurasian; in 2018, the first ASF epidemic was a chinese outbreak, followed by a subsequent outbreak in surrounding countries. ASF has become the most serious infectious disease jeopardizing the global pig industry, causing serious economic losses.
Vaccines and antiviral drugs are effective means for preventing and controlling viral diseases. For example, chinese patent application CN110093324A discloses an attenuated ASFV with gene deletion and application thereof as a vaccine, and adopts an ASF Chinese epidemic strain Pic/CN/HLJ/2018 to delete virulence genes of the ASFV by a genetic engineering technology, so that a gene deletion virus with MGF360-505R deletion and combined deletion of CD2V and MGF360-505R is obtained, and the attenuated ASF Chinese epidemic strain is further used as the vaccine to carry out immune protection of up to 100%; similarly, chinese patent application CN114107228A discloses an attenuated ASFV vaccine strain with twelve genes deleted and a vaccine containing the vaccine strain, which prepares the corresponding ASFV vaccine by attenuating the chinese epidemic strain ASFV CN/GS 2018 strain. However, for various reasons such as safety and virulence return, the current ASF attenuated vaccine is not approved to be marketed in china, and along with continuous recombination and evolution of viruses, the attenuated vaccine may lose immune protection effect on the new variant subtype ASFV. In addition to the vaccine, chinese patent application CN113797184A discloses application of Rhein in preparing medicines for preventing and treating ASFV, and experiments prove that the Rhein can effectively inhibit infection and proliferation of ASFV in cells under the condition of being lower than the maximum nontoxic dosage. However, few studies on drugs for preventing and treating ASF are reported at present, and no drugs with an anti-ASFV effect clinically exist at present. Therefore, there is an urgent need for drug development against ASFV.
Disclosure of Invention
The invention aims to solve the technical problem that the existing anti-ASFV medicine is lack and is not available, and provides an application of dibenzyl isoquinoline alkaloid in preparing the anti-ASFV medicine.
Therefore, the invention aims to provide an application of dibenzyl isoquinoline alkaloid in preparing medicines for preventing and treating ASF.
The above object of the present invention is achieved by the following technical scheme:
the bisbenzylisoquinoline alkaloid (Bisbenzylisoquinolie alkaloid) is a large class of naturally occurring alkaloids with wide distribution range in plant kingdom, variable structure types and various physiological activities, and exists in plants of Ranunculaceae, fangkefir, berberidaceae, magnoliaceae, annonaceae and the like. The bisbenzylisoquinoline alkaloid is formed by connecting two benzylisoquinoline units through an oxygen bridge, and is divided into 26 structural types according to the number of alkaloid ether bonds, substitution conditions and different connecting positions, and the common bisbenzylisoquinoline alkaloid comprises the following components: plumula Nelumbinis alkali (Liensinine), isoliensinine (Isoliensinine), neferine (Neferine), dauricine (Dauricine), and radix Puerariae; stephanine (Cepharanthine), tetrandrine (Tetrandrine), fangchinoline (Fangchinoline), berbamine (Berbamine), and the like. The prior pharmacological study shows that the bisbenzylisoquinoline alkaloid has the effects of calcium ion antagonism, blood pressure reduction, cancer resistance, nerve protection, sedation, osteoporosis resistance, allergy resistance, inflammation resistance, oxidation resistance and the like.
The inventor finds that the dibenzyl isoquinoline alkaloid is: the plumula Nelumbinis, isoliensinine, plumula Nelumbinis, dauricine, stephanine, tetrandrine, fangchinoline and berbamine can remarkably reduce ASFV-p30 protein synthesis and ASFV-B646L gene replication, and remarkably inhibit ASFV proliferation in target cell Pig Alveolar Macrophages (PAMs). In addition, the dibenzyl isoquinoline alkaloid is derived from common traditional Chinese medicines, and has wide sources and low cost. Meanwhile, the dibenzyl isoquinoline alkaloid has small toxic effect on normal cells, good selection index and high safety, and has remarkable antiviral activity on ASFV (ASFV) under the condition of no cytotoxicity concentration.
Therefore, the invention claims the application of the dibenzyl isoquinoline alkaloid in preparing the anti-ASFV drugs, wherein the dibenzyl isoquinoline alkaloid is liensinine, isoliensinine, methyl liensinine, dauricine, stephanine, tetrandrine and berbamine.
Specifically, the dibenzyl isoquinoline alkaloid has any one structure of the following formulas (I) to (VIII):
further, the dibenzyl isoquinoline alkaloid further comprises pharmaceutically acceptable salts, solvates, isomers or esters of the liensinine, isoliensinine, methylliensinine, dauricine, stephanine, tetrandrine, fangchinoline or berbamine.
In the present invention, the term "pharmaceutically acceptable salt" generally refers to any salt that is physiologically tolerable (generally, meaning that it is non-toxic) when used in a suitable manner for treatment, particularly when applied or used in humans and/or mammals. These physiologically acceptable salts may be formed with cations or bases and in the context of the present invention, in particular when administered in humans and/or mammals, they are understood to be salts formed from at least one compound provided according to the present invention, typically an acid (deprotonated), such as an anion and at least one physiologically tolerated cation, preferably an inorganic cation. These physiologically acceptable salts can also be formed with anions or acids and in the context of the present invention, in particular when administered in humans and/or mammals, they are understood to be salts formed from at least one compound provided according to the present invention, usually protonated (e.g. on nitrogen), such as a cation and at least one physiologically tolerable anion. In the context of the present invention, salts formed with physiologically tolerable acids, i.e. salts of the specific active compounds with physiologically tolerable organic or inorganic acids, may be included in particular, but not limited to, salts with hydrochloric acid, hydrobromic acid, sulfuric acid, methanesulfonic acid, formic acid, acetic acid, oxalic acid, succinic acid, malic acid, tartaric acid, mandelic acid, fumaric acid, lactic acid or citric acid.
In the present invention, the term "solvate" generally refers to a substance obtained by non-covalent bonding of any form of an active compound according to the invention to another molecule (usually a polar solvent), and may specifically include, but is not limited to, hydrates and alcoholates, such as methanolate.
Still further, the dibenzyl isoquinoline alkaloid inhibits replication of ASFV.
Further, the dibenzyl isoquinoline alkaloid has a half-effective concentration (EC 50 ) 0.31 to 2.34 mu M.
Further, the Selectivity Index (SI) of the dibenzyl isoquinoline alkaloid is 15.58-65.90.
Further, the medicament also comprises pharmaceutically acceptable auxiliary materials.
Preferably, the auxiliary materials comprise excipients, disintegrants, binders, lubricants, flavoring agents, colorants, emulsifiers or diluents.
Further, the medicament is an injection preparation, an oral preparation, an aerosol inhalation preparation or a transdermal preparation.
In addition, the invention also claims the application of the dibenzyl isoquinoline alkaloid in preparing medicines for preventing and treating ASF, wherein the dibenzyl isoquinoline alkaloid is liensinine, isoliensinine, methyl liensinine, dauricine, stephanine, tetrandrine, fangchinoline or berbamine.
Further, the dibenzyl isoquinoline alkaloid further comprises pharmaceutically acceptable salts, solvates, isomers or esters of the liensinine, isoliensinine, methylliensinine, dauricine, stephanine, tetrandrine, fangchinoline or berbamine.
The invention has the following beneficial effects:
the invention discloses a dibenzyl isoquinoline alkaloid: the plumula Nelumbinis, isoliensinine, plumula Nelumbinis, dauricine, stephanine, tetrandrine, fangchinoline and berbamine can remarkably reduce ASFV-p30 protein synthesis and ASFV-B646L gene replication, inhibit virus proliferation in PAMs, and achieve remarkable antiviral effect. In addition, the dibenzyl isoquinoline alkaloid has small toxic effect on normal cells, good selection index, obvious antiviral activity on ASFV at a concentration without cytotoxicity and high safety; in addition, the dibenzyl isoquinoline alkaloid is derived from common traditional Chinese medicines, has wide sources and low cost, and is very suitable for developing medicines for preventing and treating ASF.
Drawings
FIG. 1 is a statistical chart showing the results of experiments for inhibiting ASFV-p30 protein synthesis in PAMs using IFA to analyze different concentrations of bisbenzylisoquinoline alkaloid in example 2.
FIG. 2 is a statistical graph of the results of experiments for determining the inhibition of ASFV-B646L gene replication in PAMs using real-time fluorescent quantitative PCR for different concentrations of bisbenzylisoquinoline alkaloids in example 3.
Detailed Description
The invention is further illustrated in the following drawings and specific examples, which are not intended to limit the invention in any way. Unless specifically stated otherwise, the reagents, methods and apparatus employed in the present invention are those conventional in the art.
Reagents and materials used in the following examples are commercially available unless otherwise specified.
Example 1: half-cell toxicity (CC) of 8 bisbenzylisoquinoline alkaloids to PAMs 50 ) Half-effective concentration of anti-ASFV activity (EC 50 ) Selection Index (SI)
1. Half-cell toxicity (CC) of dibenzyl isoquinoline alkaloid to PAMs 50 ) Measurement
Resuscitates Pig Alveolar Macrophages (PAMs) (obtained by killing pig and collecting lung, and storing in liquid nitrogen tank in advance), re-suspend cells with RPMI-1640 culture solution (containing 10% foetal calf serum, 100U/mL penicillin, 100U/mL streptomycin), and concentrating 2×10 5 PAMs were seeded in 96-well plates at 100. Mu.L per well, 37℃and 5% CO 2 Culturing in a cell culture incubator. After 6h of incubation, the test compound was diluted with 1640 maintenance solution ratio containing 2% FBS, 8 dibenzyl isoquinoline alkaloid drug groups were set up with a concentration gradient of 5 total of 1.1 to 90. Mu.M, a solvent control group containing 3% DMSO and a blank control group, 100. Mu.L per well, at 37℃and 5% CO 2 After 48h incubation in a constant temperature incubator, the supernatant was discarded, and 100. Mu.L per well of MTT solution, 0.5mg/mL, was added and incubated at 37℃in the absence of light. After 4 hours, stopping incubation, discarding the supernatant, adding 150 mu L of DMSO into each well, and oscillating for 10 minutes by a low-speed oscillator to fully dissolve formazan crystals; the OD was measured at 570nm using a full wavelength microplate reader and the cell viability was calculated. Half-cell Cytotoxicity Concentration (CC) was calculated by nonlinear regression function using GraphPad Prism 9.0 software 50 )。
The calculation formula is as follows:
2. dibenzyl isoquinoline alkaloid has half the Effective Concentration (EC) of anti-ASFV activity on PAMs 50 ) Measurement
Resuscitates PAMs as above, cells were resuspended in RPMI-1640 medium and plated at 5X 10 5 Individual/well inoculates in 48-well plates; setting cell control group and virus group, each hole of 100 μl, 37deg.C, 5% CO 2 Culturing in a cell culture incubator. After 6h of culture, PBS is used for washing for 2 times, the virus group is infected with PAMs in a 48-well plate by ASFV with the concentration of 10MOI, after 2h of infection, the supernatant virus liquid is sucked, and the maintenance liquid (RPMI-1640 culture solution containing 2% of fetal calf serum) is respectively added for dilution to prepare the dibenzyl isoquinoline alkaloids with different concentrations, and the mixture is placed at 37 ℃ and 5% CO 2 After the incubator continues to incubate for 48 hoursThe culture was terminated. IFA detection was performed with 4% paraformaldehyde fixation and was observed using a fluorescent inverted microscope and recorded with photographs. Fluorescence intensity (blue fluorescence and green fluorescence) of each well was quantified using Image J software, DMSO-treated virus control group was set to 100%, and each of the other groups was compared with DMSO-treated group, and half-Effective Concentration (EC) was determined by the quantified cytoprotection rate of the drug-treated group 50 ) Values were calculated by nonlinear regression function using GraphPad Prism 9.0 software. The calculation formula is as follows:
3. bisbenzylisoquinoline alkaloid cell Selection Index (SI) against ASFV
Based on the median cytotoxicity concentration (CC 50 ) And half-effective concentration of dibenzyl isoquinoline alkaloid at PAMs against ASFV activity (EC 50 ) Calculating a Selection Index (SI), wherein the calculation formula is as follows: si=cc 50 /EC 50 . The results are shown in Table 1.
TABLE 1 CC for bis-benzylisoquinoline alkaloids 50 And EC (EC) 50 SI (service instruction) and SI (service instruction)
| Compounds of formula (I) | CC 50 (μM) | EC 50 (μM) | SI |
| Nelumbo nucifera Gaertn alkali | 36.39 | 1.5 | 24.26 |
| Isomelanin | 26.16 | 1.17 | 22.36 |
| Nelumbo nucifera Gaertn base | 36.46 | 2.34 | 15.58 |
| Dauricine | 18.28 | 0.79 | 23.14 |
| Cepharanthine | 33.04 | 1.03 | 32.07 |
| Tetrandrine powder | 62.83 | 1.38 | 45.52 |
| Fanghexiline base | 34.6 | 1.4 | 24.71 |
| Berberine amine | 20.43 | 0.31 | 65.9 |
The table shows that the 8 dibenzyl isoquinoline alkaloids have remarkable antiviral activity on ASFV at the concentration without cytotoxicity, and the SI is between 15.58 and 65.90; the dibenzyl isoquinoline alkaloid has obvious antiviral activity on ASFV under the non-cytotoxicity concentration.
Example 2: immunofluorescence method for evaluating inhibition effect of dibenzyl isoquinoline alkaloids with different concentrations on ASFV-p30 protein synthesis in PAMs (PAMs)
PAMs were resuscitated by the method of reference example 1, cells were resuspended in RPMI-1640 medium and grown at 5X 10 5 Individual/well inoculates in 48-well plates; setting a cell control group (Mock group, no test drug, no ASFV), a virus group (ASFV group, ASFV, no test drug), and a bisbenzylisoquinoline alkaloid group with different concentrations, wherein each group is repeated for 3 times; 37 ℃ and 5% CO 2 Culturing in a cell incubator for 6 hours; infection of PAMs in 48 well plate with ASFV of 10MOI for 2 hr, sucking the supernatant, adding 8 dibenzyl isoquinoline alkaloids with different concentrations diluted by maintenance solution (RPMI-1640 culture solution containing 2% fetal bovine serum), placing at 37deg.C and 5% CO 2 The incubator was incubated for a further 48 hours and the incubation was terminated. Removing supernatant, adding 400 mu L of paraformaldehyde with the mass concentration of 4% into each hole, and fixing for 30min; PBS is washed for 3 times, 200 mu L of Triton X-100 with mass concentration of 0.25% is added for membrane permeation, and the membrane is treated for 30 minutes at room temperature; adding 5% BSA blocking hybrid protein, and blocking for 1h at room temperature; PBS was washed 3 times, murine ASFV-p30 mab (1:5000 dilution) was added and incubated overnight at 4 ℃; PBS was washed 3 times and secondary antibody (Alexa was added in the dark488-labeled anti-mouse IgG, 1:1000), incubated at 37 ℃ in the dark for 1h, washed with PBS; when observed under a fluorescence microscope, green fluorescence represents ASFV-p30 protein.
The test results are shown in fig. 1, and the graph shows that 8 dibenzyl isoquinoline alkaloids have obvious inhibition effect on ASFV-p30 protein synthesis at the concentration without cytotoxicity, and have obvious dose-dependent relationship, so that the dibenzyl isoquinoline alkaloids have obvious inhibition effect on ASFV protein synthesis at the concentration without cytotoxicity.
Example 3: evaluation of the inhibition of ASFV-B646L Gene replication in PAMs by Bisbenzylisoquinoline alkaloids at different concentrations by fluorescent quantitative PCR
Resuscitates PAMs, incubates ASFV, aspirates supernatant, washes 2 times with PBS, adds different concentrations of dibenzyl isoquinoline alkaloids diluted by the maintenance solution 2 hours after challenge, simultaneously sets Mock (no test drug added, no ASFV added) and DMSO (ASFV added, no test drug added) groups, three parallel per test time point; and placed at 37℃in 5% CO 2 The incubator continued to incubate for 48 hours and then the incubation was terminated. Repeated freeze thawing is carried out for three times, DNA is extracted by using a DAN extraction kit (Nanjinouzan biotechnology Co., ltd.), and primers B646L-F and B646L-R are used for carrying out fluorescent quantitative PCR to detect CT values of B646L genes, and the influence of different concentrations of dibenzyl isoquinoline alkaloids in PAMs on ASFV replication is evaluated.
Wherein, the upstream and downstream primer sequences of the ASFV UPL:
B646L-F:5’-CCCAGGRGATAAAATGACTG-3’
B646L-R:5’-CACTRGTTCCCTCCACCGATA-3’
the test results are shown in fig. 2, where P <0.05 compared to the virus infected control group; * P <0.01; p <0.001, indicating significance of difference; ns represents no difference significance. Compared with a DMSO virus control group, the 8 dibenzyl isoquinoline alkaloids reduce the copy number of the ASFV-B646L gene under the non-cytotoxicity concentration, and have obvious dose dependency relationship; the dibenzyl isoquinoline alkaloid has an inhibiting effect on the replication of ASFV DAN under the non-cytotoxicity concentration.
The above examples are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above examples, and any other changes, modifications, substitutions, combinations, and simplifications that do not depart from the spirit and principle of the present invention should be made in the equivalent manner, and the embodiments are included in the protection scope of the present invention.
Claims (6)
1. Application of dauricine or pharmaceutically acceptable salt thereof in preparing medicine for resisting African swine fever virus is provided.
2. The use according to claim 1, wherein the pharmaceutically acceptable salt is a salt of the dauricine with hydrochloric acid, hydrobromic acid, sulfuric acid, methanesulfonic acid, formic acid, acetic acid, oxalic acid, succinic acid, malic acid, tartaric acid, mandelic acid, fumaric acid, lactic acid or citric acid.
3. The use according to claim 1, wherein the dauricine inhibits replication of african swine fever virus.
4. The use according to any one of claims 1 to 3, wherein the half-effective concentration of dauricine against african swine fever virus is 0.79 μm.
5. The use according to claim 1, wherein the medicament further comprises pharmaceutically acceptable excipients.
6. The use according to claim 1, wherein the medicament is an injectable formulation, an oral formulation, an aerosol inhalation formulation or a transdermal formulation.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0622781A (en) * | 1992-07-07 | 1994-02-01 | Kao Corp | Production of dauricine |
| CN1440984A (en) * | 2003-03-28 | 2003-09-10 | 吉林宏达科技开发有限公司 | Dauricine polysaccharide, its extraction method and the use of medicine with the compound as active component |
| CN105168215A (en) * | 2015-09-23 | 2015-12-23 | 浙江大学 | Benzylisoquinoline alkaloid and anti-tumor application of derivative thereof |
-
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0622781A (en) * | 1992-07-07 | 1994-02-01 | Kao Corp | Production of dauricine |
| CN1440984A (en) * | 2003-03-28 | 2003-09-10 | 吉林宏达科技开发有限公司 | Dauricine polysaccharide, its extraction method and the use of medicine with the compound as active component |
| CN105168215A (en) * | 2015-09-23 | 2015-12-23 | 浙江大学 | Benzylisoquinoline alkaloid and anti-tumor application of derivative thereof |
Non-Patent Citations (2)
| Title |
|---|
| Antiviral drugs targeting endosomal membrane proteins inhibit distant animal and human pathogenic viruses;I. Galindo等;《Antiviral Research》;第186卷;1-12 * |
| 中西医结合治疗咽炎临床观察;李守珍等;《现代中西医结合杂志》;第9卷(第6期);514-515 * |
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