CN108478549A - A kind of composition and its application in preparing anti-inflammatory drug - Google Patents
A kind of composition and its application in preparing anti-inflammatory drug Download PDFInfo
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- CN108478549A CN108478549A CN201810497584.6A CN201810497584A CN108478549A CN 108478549 A CN108478549 A CN 108478549A CN 201810497584 A CN201810497584 A CN 201810497584A CN 108478549 A CN108478549 A CN 108478549A
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- composition
- inflammatory
- drug
- inflammatory drug
- resveratrol
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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
- A61K31/045—Hydroxy compounds, e.g. alcohols; Salts thereof, e.g. alcoholates
- A61K31/05—Phenols
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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
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
- A61K31/7048—Compounds having saccharide radicals and heterocyclic rings having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin, digitoxin or digoxin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
Abstract
The invention belongs to drug research and development field more particularly to a kind of composition and its applications in preparing anti-inflammatory drug.Invention provides a kind of composition, and the raw material of the composition includes：Resveratrol and eriocitrin.The present invention also provides a kind of application of composition including described in any of the above one in preparing anti-inflammatory drug.In technical solution provided by the invention, resveratrol and eriocitrin generate collaboration anti-inflammatory effect, can greatly reduce the dosage of resveratrol and eriocitrin, and different Drug combinations can reach the antiphlogistic effects that multiple target point inhibits more；It can further be obtained through measuring, adverse reaction significantly mitigates, and inflammation can be effectively relieved.A kind of composition provided by the invention and its application in preparing anti-inflammatory drug, solve in the prior art, and there is the technological deficiencies of adverse reaction for anti-inflammatory drug.
Description
Technical field
The invention belongs to drug research and development field more particularly to a kind of composition and its applications in preparing anti-inflammatory drug.
Background technology
Inflammation is exactly " inflammation " usually described in people, is a kind of defense reaction of the body for stimulation, show as it is red,
Swollen, heat, pain and dysfunction.Inflammation is the immune response that self-defensive system generates.Currently, being directed to the research of inflammatory reaction
Through the big focus and emphasis as present age research.Inflammatory reaction be body under the conditions of by extraneous pessimal stimulation it is a kind of from
Suitable physiological reaction, usually there are two big stages, one acute inflammation and chronic inflammation, acute inflammation is the primary rank of inflammatory reaction
Section, is caused by innate immune system, its duration is short, and is beneficial to body., whereas if inflammation continues
For quite a long time, the second stage of inflammatory reaction, i.e. chronic inflammation will be caused, so as to cause many chronic diseases
Occur.
At the same time, inflammation can also cause the incidence of extensive physiology and pathology.Research finds, many diseases such as artery
Atherosis, diabetes, hypertension, rheumatoid arthritis, osteoarthritis, Crohn disease, tumour etc. are all to a certain extent
There are chronic inflammatory state, such as proinflammatory cytokine raising, depend on the increase of developed by molecule.It is this to continue the chronic of minuent
Inflammatory reaction promotes the pathological reaction of such disease and the interaction of the two, the exacerbation so as to cause disease or complication
Generation.
Be directed to the anti-inflammatory drug of inflammation, the adverse reaction that can effectively avoid above-mentioned physiology and pathologic conditions include infection or
Tissue damage etc..However, although traditional anti-inflammatory drug has significant effect to the inhibition of inflammation, all there are some not
Good reaction limits the use of anti-inflammatory drug.
Therefore, a kind of composition and its application in preparing anti-inflammatory drug are developed, in the prior art, is resisted for solving
There is the technological deficiencies of adverse reaction for scorching drug, become those skilled in the art's urgent problem to be solved.
Invention content
In view of this, the application the present invention provides a kind of composition and its in preparing anti-inflammatory drug, existing for solving
Have in technology, there is the technological deficiencies of adverse reaction for anti-inflammatory drug.
The present invention provides a kind of composition, the raw material of the composition includes：Resveratrol and eriocitrin.
Preferably, in terms of molar part, the raw material of the composition includes：10~20 parts of 1 part of resveratrol and eriocitrin.
The present invention also provides a kind of application of composition including described in any of the above one in preparing anti-inflammatory drug.
In conclusion invention provides a kind of composition, the raw material of the composition includes：Resveratrol and mountain balsam time
Glycosides.The present invention also provides a kind of application of composition including described in any of the above one in preparing anti-inflammatory drug.This hair
In the technical solution of bright offer, resveratrol and eriocitrin generate collaboration anti-inflammatory effect, can greatly reduce resveratrol and sage
The dosage of careless glycosides, different Drug combinations can reach the antiphlogistic effects that multiple target point inhibits more；Further through experiment
Measurement can obtain, and adverse reaction significantly mitigates, and inflammation can both be effectively relieved, but also can mitigate adverse reaction.The present invention carries
A kind of composition supplied and its application in preparing anti-inflammatory drug, solve in the prior art, there is bad for anti-inflammatory drug
The technological deficiency of reaction.
Description of the drawings
In order to more clearly explain the embodiment of the invention or the technical proposal in the existing technology, to embodiment or will show below
There is attached drawing needed in technology description to be briefly described, it should be apparent that, the accompanying drawings in the following description is only this
The embodiment of invention for those of ordinary skill in the art without creative efforts, can also basis
The attached drawing of offer obtains other attached drawings.
Fig. 1 is that the measurement result schematic diagram of NO inhibitions is generated to the RAW264.7 of LPS inductions in embodiment 2；
Fig. 2 is in embodiment 3, to the measurement result schematic diagram of the RAW264.7 growth effects of LPS inductions；
Fig. 3 is in embodiment 4, to the measurement result schematic diagram of TNF-α inhibiting effect.
Specific implementation mode
A kind of composition provided in an embodiment of the present invention and its application in preparing anti-inflammatory drug, solve the prior art
In, there is the technological deficiencies of adverse reaction for anti-inflammatory drug.
The technical scheme in the embodiments of the invention will be clearly and completely described below, it is clear that described implementation
Example is only a part of the embodiment of the present invention, instead of all the embodiments.Based on the embodiments of the present invention, this field is common
The every other embodiment that technical staff is obtained without making creative work belongs to the model that the present invention protects
It encloses.
In order to which the present invention is described in more detail, to a kind of composition provided by the invention and its prepared with reference to embodiment
Application in anti-inflammatory drug, is specifically described.
Embodiment 1
The present embodiment is the specific embodiment for preparing resveratrol and eriocitrin product mix.
5 μm of ol resveratrols and 50 μm of ol eriocitrins mixing, obtain anti-inflammatory products 1.
5 μm of ol resveratrols and 75 μm of ol eriocitrins mixing, obtain anti-inflammatory products 2.
5 μm of ol resveratrols and 100 μm of ol eriocitrins mixing, obtain anti-inflammatory products 3.
Embodiment 2
The present embodiment is to measure the RAW264.7 of 1~3 pair of LPS induction of anti-inflammatory products obtained in embodiment 1 to generate NO suppressions
The specific embodiment of effect processed.
1, Turnover of Mouse Peritoneal Macrophages RAW264.7 500,000/ml of cell density are adjusted, are inoculated in 96 orifice plates, are added per hole
100 μ l cell suspensions, are placed in 37 DEG C, 5%CO2It is incubated in incubator.
2, after for 24 hours, culture medium is discarded, is tested.Experiment is divided into 11 groups：1st group be blank group, be added 100 μ L without
Blood serum medium；2nd group is LPS groups (lipopolysaccharides) group, and xx μ L lipopolysaccharides is added；3rd~5 group is resveratrol group, is added respectively
Enter 2.5 μM, 5 μM, 10 μM of resveratrol；6th~8 group is eriocitrin group, is separately added into 25 μM of eriocitrin, 50 μM, 100 μ
M；9th~11 group is anti-inflammatory products experimental group, is separately added into anti-inflammatory products 1~3xx μ L；In every group, Drug level has 8 holes weight
It is multiple.Dosing post-processes one hour, is then separately added into the stimulation of 10 μ g/mlLPS solution for the 1st group and the 2nd group.LPS is added to be placed on
37 DEG C, 5%CO2It is incubated in incubator.
3, after for 24 hours, 50 μ L supernatants is taken to be added in 96 well culture plates per hole, 50 μ LGriessA is then added per hole
Reagent and 50 μ LGriessB reagents, gently shake culture plate mixing, are placed in incubation at room temperature.
4, after 10min, with the absorption photometric value at enzyme mark photometer detection 540nm wavelength, the concentration of NO is according to NaNO2Mark
Directrix curve is calculated.The percentage that the result of drug study group is scaled LPS control groups indicates.This is because NO is gaseous state
Endogenous biological messenger molecule can be synthesized by many cells, and NO can participate in diversified physiological and biochemical procedure, such as：
Blood vessel dilatation, macrophage-mediated cytotoxicity etc..NO is referred to as nitric oxide by a series of Enzyme catalyzed synthesis, these enzymes
Synzyme；Under inflammatory conditions, nitric oxide synthetase generates NO by high level activation；NO enters after generating in culture solution, becomes
NO2-Ion therefore can be by measuring NO2-, reflect the content of NO.
Experimental results please refer to Fig.1.
As can be drawn from Figure 1, the RAW264.7 that veratryl alcohol and eriocitrin independent medication induce LPS in low dosage
The NO of generation influences smaller, and when the two drug combination significantly suppresses nitric oxide production generation.
Embodiment 3
The present embodiment is that anti-inflammatory products 1~3 and dexamethasone (Dexamethasone) obtained are right in measurement embodiment 1
The specific embodiment of the RAW264.7 growth effects of LPS inductions.
By RAW264.7 cells, got off with trypsin digestion, is added in the DMEM culture mediums containing 10%FBS and is made carefully
Born of the same parents' suspension.With inoculum density 3 × 104The concentration of a/mL is inoculated in 96 orifice plates (100 holes μ L/).
Experiment packet is same as Example 2, while blank zeroing hole (culture medium bed board hole) is arranged.In 5%CO2、37℃
After overnight incubation, culture solution is carefully sucked out, the MTT solution (0.5mg/mL) that 150 μ L serum free mediums are prepared is added per hole；After
After continuous culture 4h, culture solution in hole is carefully sucked.200 μ LDMSO are added per hole, ten minutes are stood in incubator；It is surveyed with microplate reader
Measure the light absorption value in each hole at 570nm.
Experimental results please refer to Fig. 2.
As can be drawn from Figure 2, anti-inflammatory products 1~3 do not interfere with the normal growth of cell, and with anti-inflammatory common drug
Sai meter Song is compared, and natural products group does not inhibit the growth of macrophage.
Embodiment 4
The present embodiment is the specific embodiment for measuring 1~3 pair of TNF-α inhibiting effect of anti-inflammatory products obtained in embodiment 1.
Turnover of Mouse Peritoneal Macrophages RAW264.7 500,000/ml of density are adjusted, are inoculated in 35mm capsules, add 2ml per ware
Cell suspension is placed in 37 DEG C, 5%CO2It is incubated in incubator.
After for 24 hours, culture medium is discarded, experiment packet is same as Example 2, pre-processes one hour.Then 2 difference of group 1 and group
The stimulation of 10 μ g/mlLPS solution is added, each Drug level has the repetition of 8 holes.After adding LPS, it is placed in 37 DEG C, 5%CO2Incubator
Interior incubation.
After for 24 hours, 100 μ L supernatants, -80 DEG C of freezen protectives are taken per hole.
TNF-α (tumor necrosis factor) ELISA kit is taken out from 4 DEG C of refrigerators, equilibrium at room temperature 30min is ready to test
Required reagent.
Sample-adding：Gauge orifice, sample to be tested hole are set respectively.Add 100 μ L of standard items or sample to be tested respectively per hole, gently shakes
It shakes, is covered with plate patch, 37 DEG C are incubated 2 hours.
Liquid is discarded, is dried, washing is not had to.
Add 100 μ L of biotin labelled antibodies working solution per hole, is covered with new plate patch, 37 DEG C are incubated 1 hour.
Liquid is discarded, is dried, board-washing 3 times.It impregnates two minutes every time, 200 μ L/ are per hole, drying.
Add 100 μ L of Horseradish peroxidase-conjugated avidin working solution per hole, is covered with new plate patch, 37 DEG C incubate 1 hour.
Liquid is discarded, is dried, board-washing 5 times.It impregnates two minutes every time, 200 μ L/ are per hole, drying.
Sequentially add 90 μ L of substrate solution per hole, 37 DEG C are protected from light colour developing 15-30 minutes.
50 μ L of terminate liquid are sequentially added per hole, terminate reaction.
Sequentially measure the optical density (OD values) in each hole in 5 minutes in 450nm wavelength with microplate reader after termination of the reaction.
Acquired results please refer to Fig. 3.
It is as can be drawn from Figure 3, smaller for the inhibiting effect of TNF-α when resveratrol or eriocitrin is used alone,
When to resveratrol and eriocitrin drug combination, the inhibiting effect of TNF-α is significantly increased.
It can be obtained from above-mentioned technical proposal, technical solution provided in an embodiment of the present invention has the following advantages：
1, in technical solution provided by the invention, resveratrol and eriocitrin are used in combination, drug effect is substantially better than list
Private medicine, the inflammatory model established to LPS inducing macrophages have good inhibition.
2, in technical solution provided by the invention, dosage is low in inflammation, hence it is evident that the concentration for reducing independent medication, from
And the toxic side effect of drug alone is reduced, reduce the injury to human body.
3, technical solution price provided by the invention is relatively cheap, alleviates the financial burden of patient.
In conclusion invention provides a kind of composition, the raw material of the composition includes：Resveratrol and mountain balsam time
Glycosides.The present invention also provides a kind of application of composition including described in any of the above one in preparing anti-inflammatory drug.This hair
In the technical solution of bright offer, resveratrol and eriocitrin generate collaboration anti-inflammatory effect, can greatly reduce resveratrol and sage
The dosage of careless glycosides, different Drug combinations can reach the antiphlogistic effects that multiple target point inhibits more；Further through experiment
Measurement can obtain, and adverse reaction significantly mitigates, and inflammation can both be effectively relieved, but also can mitigate adverse reaction.The present invention carries
A kind of composition supplied and its application in preparing anti-inflammatory drug, solve in the prior art, there is bad for anti-inflammatory drug
The technological deficiency of reaction.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also answered
It is considered as protection scope of the present invention.
Claims (3)
1. a kind of composition, which is characterized in that the raw material of the composition includes：Resveratrol and eriocitrin.
2. composition according to claim 1, which is characterized in that in terms of molar part, the raw material of the composition includes：In vain
10~20 parts of 1 part of veratryl alcohol and eriocitrin.
3. a kind of application including composition as claimed in claim 1 or 2 in preparing anti-inflammatory drug.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810497584.6A CN108478549A (en) | 2018-05-22 | 2018-05-22 | A kind of composition and its application in preparing anti-inflammatory drug |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201810497584.6A CN108478549A (en) | 2018-05-22 | 2018-05-22 | A kind of composition and its application in preparing anti-inflammatory drug |
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| CN108478549A true CN108478549A (en) | 2018-09-04 |
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| CN201810497584.6A Pending CN108478549A (en) | 2018-05-22 | 2018-05-22 | A kind of composition and its application in preparing anti-inflammatory drug |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2013086323A2 (en) * | 2011-12-08 | 2013-06-13 | Metaproteomics, Llc | Supplemented oil compositions and methods for improved health |
| US20170014439A1 (en) * | 2014-03-31 | 2017-01-19 | Robert M. Brewster | Flavonoid compositions and uses thereof |
-
2018
- 2018-05-22 CN CN201810497584.6A patent/CN108478549A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2013086323A2 (en) * | 2011-12-08 | 2013-06-13 | Metaproteomics, Llc | Supplemented oil compositions and methods for improved health |
| US20170014439A1 (en) * | 2014-03-31 | 2017-01-19 | Robert M. Brewster | Flavonoid compositions and uses thereof |
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Application publication date: 20180904 |
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