CN107308169A - A kind of pharmaceutical composition with antivirus action - Google Patents
A kind of pharmaceutical composition with antivirus action Download PDFInfo
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- CN107308169A CN107308169A CN201710551838.3A CN201710551838A CN107308169A CN 107308169 A CN107308169 A CN 107308169A CN 201710551838 A CN201710551838 A CN 201710551838A CN 107308169 A CN107308169 A CN 107308169A
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- cinobufagin
- bufalin
- pharmaceutical composition
- antivirus action
- weight
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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
- A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
- A61K31/58—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids containing heterocyclic rings, e.g. danazol, stanozolol, pancuronium or digitogenin
- A61K31/585—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids containing heterocyclic rings, e.g. danazol, stanozolol, pancuronium or digitogenin containing lactone rings, e.g. oxandrolone, bufalin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/28—Steroids, e.g. cholesterol, bile acids or glycyrrhetinic acid
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- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- General Chemical & Material Sciences (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Engineering & Computer Science (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The present invention relates to drug field, a kind of pharmaceutical composition with antivirus action is specifically disclosed.Described pharmaceutical composition includes Bufalin and Cinobufagin；Described Bufalin and the weight ratio of Cinobufagin are 1 ~ 10:1~10.Synergistic corrosion virus effect can be played after Bufalin and Cinobufagin mixing in said composition, is 1 ~ 3 by weight especially by Bufalin and Cinobufagin:Its synergistic corrosion virus effect becomes apparent from after 1 ~ 3 mixing, and the effect of its resisiting influenza virus is suitable with positive control drug Ribavirin.
Description
Technical field
The present invention relates to drug field, and in particular to a kind of pharmaceutical composition with antivirus action.
Background technology
The dried venom of toads is the secretion of toad ear rear gland and epidermal gland, white milky liquid or pale yellow syrup, poisonous.The dried venom of toads
Slurries are extracted, are dried, are painted, can be used as medicine after sealing preserve.With detoxifying, ease pain, have one's ideas straightened out, a variety of functions such as antitumor and by
Extensive use.Toadvenom is used for clinical treatment coronary heart disease, Japan production " lifesaving is red " by raw material of the dried venom of toads by Germany.China
There is dried venom of toads composition in more than the 50 kind Chinese patent drugs such as famous Melnikov's method, toothache killer pill, heart treasured, a cinobufagin injection.
Because the dried venom of toads has very big toxicity, existing marketed drug preparation is all the dried venom of toads to be treated through specific process
What extract was used as medicine.Although toad cake extract can clinically obtain certain effect, due to complicated component therein, and
And according to the difference of extraction process and raw material sources the active constituent content in toad cake extract can be caused unstable, quality is not
Easy to control, the security of medication can also have a greatly reduced quality.Therefore, a kind of medicine of definite ingredients is developed from the dried venom of toads, with wide
Market prospects.
It is existing studies have found that, substantial amounts of arenobufagin class material, steroid, alkaloids chemical composition are contained in the dried venom of toads.
But among numerous compositions, the combination of which composition has excellent antivirus action, and the combination of which composition has
Low toxicity effect there is no research to report.Therefore, a kind of antiviral and definite ingredients composition with excellent effect is developed to people
Class health cause has great contribution, especially develops the medicine that a kind of definite ingredients, antivirus action are excellent and toxicity is low
Composition, its meaning is more great.
The content of the invention
The technical problems to be solved by the invention are to provide a kind of pharmaceutical composition with antivirus action.
Above-mentioned technical problem to be solved by this invention, is achieved by the following technical programs：
The present invention is provided, a kind of pharmaceutical composition with antivirus action, and it includes Bufalin and Cinobufagin.
Preferably, the weight ratio of described Bufalin and Cinobufagin is 1 ~ 10:1~10.
It is further preferred that described Bufalin and the weight ratio of Cinobufagin are 1 ~ 5:1~5.
It is further preferred that described Bufalin and the weight ratio of Cinobufagin are 1 ~ 3:1~3.
Inventor is had found through many experiments, and synergistic corrosion virus effect can be played after Bufalin and Cinobufagin are mixed,
It is 1 ~ 3 by weight especially by Bufalin and Cinobufagin:Its synergistic corrosion virus effect becomes apparent from after 1 ~ 3 mixing.
As it is known by the man skilled in the art that can be acted synergistically between medicine, and medicine combination and each medicine
Between consumption have close relationship；Drug regimen is improper can not only to occur synergy can press down each other on the contrary
Make and use；In addition, consumption proportion is improper between medicine, equally decide whether act synergistically between medicine or suppress to make
With.Weight proportion between drug regimen of the present invention and pharmaceutical composition is that invention finds out the tool come through many experiments
There are combination and proportioning that synergistic corrosion virus are acted on.
Preferably, the described pharmaceutical composition with antivirus action, also comprising phytosterin compound, described sterol
Class compound is selected from the mixing of one or both of cupreol and campesterol.
Preferably, the weight consumption of described phytosterin compound is the 1/10 of Bufalin and Cinobufagin weight sum
~1/30。
It is further preferred that the weight consumption of described phytosterin compound is Bufalin and Cinobufagin weight sum
1/10 ~ 1/20.
Preferably, described phytosterin compound is selected from the mixing of cupreol and campesterol, cupreol and rape oil
The weight ratio of sterol is 1 ~ 5:1~5.
It is further preferred that the weight ratio of cupreol and campesterol is 1 ~ 3:1~3.
Inventor by many experiments also further study show that, although Bufalin and Cinobufagin combination after have association
With resisiting influenza virus effect, but its cytotoxicity is still than larger, unsuitable heavy dose long-term taking；But after phytosterin compound
After addition, after especially cupreol and campesterol are added, the cytotoxicity of Bufalin and Cinobufagin composition is significantly
The antiviral effect of Bufalin and Cinobufagin is had no effect on after reduction, and cupreol and campesterol addition.
Preferably, described virus is influenza virus.
Most preferably, described influenza virus is avian influenza virus or parainfluenza virus.
Preferably, described pharmaceutical composition also includes pharmaceutically acceptable carrier.
Preferably, the formulation of described anti-inflammatory pharmaceutical compositions is liquid injection agent, powder-injection, tablet, capsule, flexible glue
Capsule, powder, pill, oral liquid, paste, granule or dressing.
Preferably, described virus is influenza virus, avian influenza virus, enterovirus, HPV, hepatitis
Poison.
Beneficial effect：The invention provides a kind of brand-new pharmaceutical composition with antivirus action, toad in composition
Synergistic corrosion virus effect can be played after malicious spirit and Cinobufagin mixing, especially by Bufalin and Cinobufagin by weight
For 1 ~ 3:Its synergistic corrosion virus effect becomes apparent from after 1 ~ 3 mixing, effect and the positive control drug Ribavirin of its resisiting influenza virus
Quite；In addition, the present invention can also add phytosterin compound, especially cupreol in the composition and campesterol is added
Afterwards, the cytotoxicity of Bufalin and Cinobufagin composition is greatly reduced.
Embodiment
The present invention is explained further below in conjunction with specific embodiment, but embodiment does not do any type of limit to the present invention
It is fixed.
The pharmaceutical composition resisiting influenza virus of embodiment 1 is tested
In 96 porocyte culture plates for covering with MDCK cell monolayers layer, 100TCID50 is added（100 times of median infective doses）Virus
Liquid（H1N1 viruses or PIV3 parainfluenza viruses）100 μ L/ holes, adsorb 1h, are then separately added into per hole, add dilute with maintaining liquid
The μ L of series concentration sample solution 100 released, are placed in 37 DEG C, are incubated in 5% CO2 incubators.By the old liquid in 96 orifice plates after 48h
Suction is abandoned, then is diluted with CCK8 solution respectively by being added per the μ L of hole 100 in tested prospect hole after 10 times, after incubator is incubated 1h, use
ELIASA surveys absorbance in 450nm, tests the IC50 values of each sample solution.
IC50 calculations are as follows：Cell survival rate=（Medicine feeding hole OD values-virus control group OD values）/（Cell controls group
OD values-virus control group OD values）×100%；Using drug concentration as X-axis, cell survival rate is Y-axis, in Origin mappings, is obtained
The corresponding concentration of 50% cell survival rate is then IC50.
Sample is respectively Bufalin；Cinobufagin；Bufalin and Cinobufagin are 10 by weight:1 composition it is mixed
Close sample；Bufalin and Cinobufagin are 1 by weight:The biased sample of 10 compositions；Bufalin and Cinobufagin are by weight
Amount is than being 3:The biased sample of 1 composition；Bufalin and Cinobufagin are 1 by weight:The biased sample of 3 compositions；Bufalin
It is 1 by weight with Cinobufagin:The biased sample of 1 composition；Bufalin and Cinobufagin are 1 by weight:1 composition
Biased sample；Bufalin, Cinobufagin and cupreol are 1 by weight:1:The biased sample of 0.05 composition；Bufalin,
Cinobufagin, cupreol and campesterol are 1 by weight:1:0.03:The biased sample of 0.02 composition；Control sample
Solution is Ribavirin.The antiviral effect test data of each sample solution is shown in Table 1.
The pharmaceutical composition resisiting influenza virus IC50 test results of the present invention of table 1.
| Sample | H1N1 virus IC50 values（μM） | PIV3 virus IC50 values（μM） |
| Bufalin | 10.53 | 15.46 |
| Cinobufagin | 15.44 | 16.89 |
| Bufalin：Cinobufagin（10:1） | 8.41 | 10.12 |
| Bufalin：Cinobufagin（1:10） | 9.88 | 11.32 |
| Bufalin：Cinobufagin（3:1） | 4.27 | 5.71 |
| Bufalin：Cinobufagin（1:3） | 4.82 | 5.43 |
| Bufalin：Cinobufagin（1:1） | 3.66 | 4.21 |
| Bufalin：Cinobufagin：Cupreol（1:1:0.05） | 3.98 | 4.33 |
| Bufalin：Cinobufagin：Cupreol：Campesterol（1:1:0.03:0.02） | 3.84 | 4.29 |
| Ribavirin | 3.41 | 4.34 |
It can be seen that Bufalin and Cinobufagin, which is used alone, has certain resisiting influenza virus from the data of table 1（Bird flu
Virus and parainfluenza virus）Effect, but its IC50 value is several times of positive control drug Ribavirin, and toad is used alone in this explanation
Malicious spirit and Cinobufagin, the effect of its resisiting influenza virus is much not as good as Ribavirin.
It is can also be seen that from the data of table 1 when Bufalin and Cinobufagin are with 10:1 or 1:Used after 10 ratio mixing,
Its IC50 value of its resisiting influenza virus is smaller when Bufalin and Cinobufagin than being used alone, and illustrates when Bufalin and the magnificent dried venom of toads
After malicious base mixing, the effect of its resisiting influenza virus can further be strengthened, and this explanation is produced after Bufalin and Cinobufagin mixing
The resisiting influenza virus effect of collaboration.It is can also be seen that from the data of table 1 when Bufalin and Cinobufagin are with 1 ~ 3:1 ~ 3 ratio
After example mixing, the IC50 values of its IC50 value and Ribavirin are approached, and this explanation works as Bufalin and Cinobufagin with 1 ~ 3:1 ~ 3
After ratio mixing, it cooperates with resisiting influenza virus effect to become apparent from.
It can also be seen that adding cupreol and campesterol etc. after Bufalin and Cinobufagin mixing from the data of table 1
Drug ingedient cooperates with resisiting influenza virus effect to have no influence on it.
In summary, because Bufalin and Cinobufagin have the effect of collaboration resisiting influenza virus, therefore can be by toad
Used after malicious spirit and Cinobufagin mixing as antiviral drugs, especially as anti-avian influenza virus and parainfluenza virus
Medicine.
The pharmaceutical composition cytotoxicity test of embodiment 2
By Hep-2 cell culture in 96 well culture plates, after cell monolayer length is good, it is serial dense that addition maintaining liquid has diluted
The μ L of sample solution 100 are spent, in 5%CO2, cultivate 3 days in 37 DEG C of incubators；Then 10 μ lMTT solution are added, MTT solution
Concentration is 5mg/mL, is configured with cushioning liquid, continues to cultivate 4 hours；Sample solution is suctioned out, dimethyl sulfoxide is added, at room temperature, will
96 orifice plates, which are placed in micro- emptying aperture plate oscillator, to be vibrated 10 minutes；The OD values in each hole are determined with ELIASA, measurement wavelength is 570nm,
Reference wavelength is 630nm, calculates median lethal toxic concentration (CC50) of the sample to cell.Every group sets 4 balance pipes, and every group real
Test and be repeated 3 times.Result of calculation, draws curve, obtains half toxic concentration (CC50).
Sample is respectively Bufalin；Cinobufagin；Bufalin and Cinobufagin are 10 by weight:1 composition it is mixed
Close sample；Bufalin and Cinobufagin are 1 by weight:The biased sample of 1 composition；Bufalin, Cinobufagin and β-paddy
Sterol is 1 by weight:1:The biased sample of 0.05 composition；Bufalin, Cinobufagin, cupreol and campesterol are by weight
Amount is than being 1:1:0.03:The biased sample of 0.02 composition；Bufalin, Cinobufagin and cupreol are 1 by weight:1:
The biased sample of 0.1 composition；Bufalin, Cinobufagin, cupreol and campesterol are 1 by weight:1:0.06:0.04
The biased sample of composition, control sample solution is virazole.The cytotoxicity test data of each sample are shown in Table 2.
The pharmaceutical composition resisiting influenza virus IC50 test results of the present invention of table 2.
| Sample | CC50 values（μM） |
| Bufalin | 25.44 |
| Cinobufagin | 18.33 |
| Bufalin：Cinobufagin（3:1） | 22.14 |
| Bufalin：Cinobufagin（1:3） | 19.18 |
| Bufalin：Cinobufagin（1:1） | 21.44 |
| Bufalin：Cinobufagin：Cupreol（1:1:0.05） | 40.13 |
| Bufalin：Cinobufagin：Cupreol：Campesterol（1:1:0.03:0.02） | 55.21 |
| Bufalin：Cinobufagin：Cupreol（1:1:0.1） | 52.36 |
| Bufalin：Cinobufagin：Cupreol：Campesterol（1:1:0.06:0.04） | 64.45 |
| Virazole | 62.5 |
As can be seen that Bufalin, Cinobufagin or combination is used alone from the data of table 2, its CC50 to cell
Value is far smaller than CC50 value of the virazole to cell, and this explanation is used alone Bufalin, Cinobufagin or combination and made
For medicine, it has certain toxicity, and its cytotoxicity is far longer than the cytotoxicity of virazole, therefore, should not big agent for a long time
Amount takes Bufalin, Cinobufagin or combination.
From table 2 it can also be seen that when in Bufalin and Cinobufagin add phytosterin compound after, Bufalin with
The cytotoxicity of Cinobufagin is neutralized, such as Bufalin：Cinobufagin：Cupreol=1:1:0.05 or 1:1:0.1
When, its CC50 value is greatly improved, and this is illustrated when phytosterin compound consumption by weight is Bufalin and Cinobufagin weight
After the 1/10 ~ 1/20 of sum is added in Bufalin and Cinobufagin, Bufalin and Cinobufagin can be mitigated to cell
Toxicity；It can also be seen that when the group that cupreol and campesterol are added in Bufalin and Cinobufagin from above-mentioned data
After conjunction, its cytotoxicity is further reduced, and its CC50 value is suitable with virazole, is added in this explanation Bufalin and Cinobufagin
After the combination for entering cupreol and campesterol, cupreol and campesterol collaboration have neutralized Bufalin and Cinobufagin
Toxicity so that its cytotoxicity is further greatly reduced.
Although the data of comprehensive Tables 1 and 2 have collaboration anti influenza after can be seen that Bufalin and Cinobufagin combination
Virus function, but its cytotoxicity is still than larger, unsuitable heavy dose long-term taking；But after being added after phytosterin compound,
After especially cupreol and campesterol are added, the cytotoxicity of Bufalin and Cinobufagin composition is greatly reduced, and
It can be seen that cupreol and campesterol have no effect on the antiviral of Bufalin and Cinobufagin after adding from the data of table 1
Effect.
Embodiment 3 has the pharmaceutical composition of antivirus action
It is 1 by weight by Bufalin and Cinobufagin:1 mixing is as active ingredient, then by the conventional side of pharmaceutical field
Powder-injection is made in method.
Embodiment 4 has the pharmaceutical composition of antivirus action
It is 10 by weight by Bufalin and Cinobufagin:1 mixing is as active ingredient, then by the conventional side of pharmaceutical field
Powder-injection is made in method.
Embodiment 5 has the pharmaceutical composition of antivirus action
It is 1 by weight by Bufalin and Cinobufagin:10 mixing are as active ingredient, then by the conventional side of pharmaceutical field
Powder-injection is made in method.
Embodiment 6 has the pharmaceutical composition of antivirus action
It is 5 by weight by Bufalin and Cinobufagin:1 mixing is as active ingredient, then by the conventional side of pharmaceutical field
Liquid injection agent is made in method.
Embodiment 7 has the pharmaceutical composition of antivirus action
It is 1 by weight by Bufalin and Cinobufagin:5 mixing are as active ingredient, then by the conventional side of pharmaceutical field
Liquid injection agent is made in method.
Embodiment 8 has the pharmaceutical composition of antivirus action
It is 3 by weight by Bufalin and Cinobufagin:1 mixing is as active ingredient, then by pharmaceutical field
Tablet is made in conventional method.
Embodiment 9 has the pharmaceutical composition of antivirus action
It is 1 by weight by Bufalin and Cinobufagin:3 mixing are as active ingredient, then by pharmaceutical field
Capsule is made in conventional method.
Embodiment 10 has the pharmaceutical composition of antivirus action
It is 1 by weight by Bufalin, Cinobufagin and cupreol:1:0.05 mixes as active ingredient,
Then tablet is made by the conventional method of pharmaceutical field.
Embodiment 11 has the pharmaceutical composition of antivirus action
It is 1 by weight by Bufalin, Cinobufagin and cupreol:1:0.1 mixes as active ingredient,
Then tablet is made by the conventional method of pharmaceutical field.
Embodiment 12 has the pharmaceutical composition of antivirus action
Bufalin, Cinobufagin, cupreol and campesterol are 1 by weight:1:0.03:0.02 mixing is made
For active ingredient, liquid injection agent then is made by the conventional method of pharmaceutical field.
Embodiment 12 has the pharmaceutical composition of antivirus action
Bufalin, Cinobufagin, cupreol and campesterol are 1 by weight:1:0.06:0.04 mixing is made
For active ingredient, liquid injection agent then is made by the conventional method of pharmaceutical field.
Claims (10)
1. a kind of pharmaceutical composition with antivirus action, it is characterised in that it includes Bufalin and Cinobufagin.
2. the pharmaceutical composition according to claim 1 with antivirus action, it is characterised in that described Bufalin and
The weight ratio of Cinobufagin is 1 ~ 10:1~10.
3. the pharmaceutical composition according to claim 1 with antivirus action, it is characterised in that described Bufalin and
The weight ratio of Cinobufagin is 1 ~ 5:1~5.
4. the pharmaceutical composition according to claim 1 with antivirus action, it is characterised in that described Bufalin and
The weight ratio of Cinobufagin is 1 ~ 3:1~3.
5. the pharmaceutical composition according to claim 1 with antivirus action, it is characterised in that also comprising sterols
Compound, described phytosterin compound is selected from the mixing of one or both of cupreol and campesterol.
6. the pharmaceutical composition according to claim 5 with antivirus action, it is characterised in that described sterols
The weight consumption of compound is the 1/10 ~ 1/30 of Bufalin and Cinobufagin weight sum；Preferably, described sterols chemical combination
The weight consumption of thing is the 1/10 ~ 1/20 of Bufalin and Cinobufagin weight sum.
7. the pharmaceutical composition according to claim 5 with antivirus action, it is characterised in that described sterols
The weight ratio of mixing of the compound selected from cupreol and campesterol, cupreol and campesterol is 1 ~ 5:1~5；Preferably,
The weight ratio of cupreol and campesterol is 1 ~ 3:1~3.
8. the pharmaceutical composition according to claim 1 with antivirus action, it is characterised in that also comprising pharmaceutically may be used
The carrier of receiving.
9. the pharmaceutical composition according to claim 1 with antivirus action, it is characterised in that described anti-inflammatory drug
The formulation of composition is liquid injection agent, powder-injection, tablet, capsule, soft capsule, powder, pill, oral liquid, paste, particle
Agent or dressing.
10. the pharmaceutical composition according to claim 1 with antivirus action, it is characterised in that described virus is
Influenza virus, avian influenza virus, enterovirus, HPV, hepatitis viruse.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201710551838.3A CN107308169A (en) | 2017-07-07 | 2017-07-07 | A kind of pharmaceutical composition with antivirus action |
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| CN201710551838.3A CN107308169A (en) | 2017-07-07 | 2017-07-07 | A kind of pharmaceutical composition with antivirus action |
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| CN107308169A true CN107308169A (en) | 2017-11-03 |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107913300A (en) * | 2018-01-19 | 2018-04-17 | 河南牧农生物科技有限公司 | A kind of formula for treating poultry influenza ewcastle disease |
| CN111437293A (en) * | 2020-04-24 | 2020-07-24 | 江苏省中医药研究院 | Application of venenum Bufonis extract in preparing medicine for treating infectious diseases caused by SARS-CoV-2 virus |
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| CN103191133A (en) * | 2013-04-23 | 2013-07-10 | 山东中医药大学 | Preparation method for extracting cinobufagin and bufalin mixture from venenum bufonis |
| CN105560254A (en) * | 2014-10-09 | 2016-05-11 | 吉林大学 | Novel use of cinobufagin and resibufogenin in inhibition of enterovirus type 71 infection |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103191133A (en) * | 2013-04-23 | 2013-07-10 | 山东中医药大学 | Preparation method for extracting cinobufagin and bufalin mixture from venenum bufonis |
| CN105560254A (en) * | 2014-10-09 | 2016-05-11 | 吉林大学 | Novel use of cinobufagin and resibufogenin in inhibition of enterovirus type 71 infection |
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| XIAOYAN CUI 等: "Anti-hepatitis B Virus Activities of Cinobufacini and Its Active Components Bufalin and Cinobufagin in HepG2.2.15 Cells", 《BIOL. PHARM. BULL. 》 * |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107913300A (en) * | 2018-01-19 | 2018-04-17 | 河南牧农生物科技有限公司 | A kind of formula for treating poultry influenza ewcastle disease |
| CN111437293A (en) * | 2020-04-24 | 2020-07-24 | 江苏省中医药研究院 | Application of venenum Bufonis extract in preparing medicine for treating infectious diseases caused by SARS-CoV-2 virus |
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Application publication date: 20171103 |