CN114432430A - Pharmaceutical composition containing scutellarin, baicalin and/or breviscapine and application thereof - Google Patents
Pharmaceutical composition containing scutellarin, baicalin and/or breviscapine and application thereof Download PDFInfo
- Publication number
- CN114432430A CN114432430A CN202011191253.3A CN202011191253A CN114432430A CN 114432430 A CN114432430 A CN 114432430A CN 202011191253 A CN202011191253 A CN 202011191253A CN 114432430 A CN114432430 A CN 114432430A
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- CN
- China
- Prior art keywords
- growth factor
- scutellarin
- vascular endothelial
- medicament
- pharmaceutical composition
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
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Abstract
The invention provides a pharmaceutical composition containing scutellarin and analogues thereof and application thereof. The pharmaceutical composition comprises vascular endothelial growth factor and scutellarin and/or analogues thereof, wherein the analogues are selected from one or more of baicalin and breviscapine. The invention provides a new application of scutellarin and analogues thereof in promoting wound healing and repair, which can be used for synergically increasing the effects of Vascular Endothelial Growth Factor (VEGF) and platelet growth factor group (SGC) in the aspect, so that the scutellarin and the analogues thereof can be used as medicines or cosmetics for treating and/or relieving skin wounds and/or scars, and have good clinical application prospect and market prospect.
Description
Technical Field
The invention belongs to the technical field of medicines, and particularly relates to a pharmaceutical composition containing scutellarin and analogues thereof and application thereof.
Background
The skin is the main barrier between the human body and the environment. It is known to have many protective functions, including preventing the body from losing water, preventing bacteria and toxic substances from outside the environment, etc., and therefore, it is very important to maintain the integrity and continuity of the skin.
Skin injury can stimulate a wound healing response. In the wound healing reaction, the stages of blood clot formation, inflammation, re-epithelialization, etc. are coordinated with each other, so that the skin is restored to a normal state as soon as possible. At these various stages, re-epithelialization is crucial, with keratinocyte migration serving a central role. Migration of keratinocytes is a complex process involving the action of multiple biomolecules.
When the skin is injured, platelets act as a first-line defense guard, immediately secreting various growth factors to promote wound healing, such as Epidermal Growth Factor (EGF), Vascular Endothelial Growth Factor (VEGF), Fibroblast Growth Factor (FGF), platelet-derived growth factor (PDGF), and the like. These growth factors play an important role in regulating the contraction process of transitional keratinocytes in the wound bed.
The development of effective skin wound healing agents is advantageous for promoting wound healing.
Disclosure of Invention
In order to solve the problems in the prior art, the invention provides a pharmaceutical composition containing scutellarin and analogues thereof and application thereof.
Specifically, the present invention provides:
(1) a pharmaceutical composition, comprising vascular endothelial growth factor and scutellarin and/or analogues thereof, wherein the analogues are selected from one or more of baicalin and breviscapine.
(2) The pharmaceutical composition of (1), wherein the molar ratio of the VEGF to the scutellarin or the analogues thereof is 1 (4000-12000).
(3) The pharmaceutical composition of (1), wherein the pharmaceutical composition comprises a population of platelet growth factors.
(4) The pharmaceutical composition according to (3), wherein the molar ratio of the vascular endothelial growth factor to the scutellarin or the analogues thereof in the platelet growth factor group is 1 (30000-150000).
(5) Application of scutellarin and analogues thereof in preparing medicines or skin care products for promoting effects of vascular endothelial growth factor and/or platelet growth factor groups, wherein the analogues are selected from one or more of baicalin and breviscapine.
(5) The use of (5), wherein the effects include promoting keratinocyte migration, promoting wound repair and/or healing, promoting matrix metalloproteinase expression, activating vascular endothelial growth factor-specific receptor 2 and its mediated MAPK signaling pathway.
(7) The use according to (5), wherein the medicament is a medicament for the treatment and/or alleviation of skin wounds and/or scars.
(8) The use according to (5), wherein the skin care product is a skin care product for treating and/or alleviating skin wounds and/or scars.
(9) Use of a pharmaceutical composition according to any one of (1) to (4) for the preparation of a medicament or a skin care product for promoting the action of vascular endothelial growth factor and/or platelet growth factor groups.
(10) The use of (9), wherein the effects comprise promoting keratinocyte migration, promoting wound repair and/or healing, promoting matrix metalloproteinase expression, activating vascular endothelial growth factor-specific receptor 2 and its mediated MAPK signaling pathway.
(11) The use according to (9), wherein the medicament is a medicament for the treatment and/or alleviation of skin wounds and/or scars.
(12) The use according to (9), wherein the skin care product is a skin care product for treating and/or alleviating skin wounds and/or scars.
(13) A medicament for promoting the action of a group of vascular endothelial growth factor and/or platelet growth factor, comprising the pharmaceutical composition according to any one of (1) to (4).
(14) The medicament of (13), wherein the medicament further comprises a pharmaceutically acceptable excipient.
(15) The medicament according to (13), wherein the medicament is in the form of a liniment or an injection.
(16) A skin care product for promoting the action of a group of vascular endothelial growth factor and/or platelet growth factor, comprising the pharmaceutical composition according to any one of (1) to (4).
(17) The skin care preparation according to (16), wherein the skin care preparation further comprises one or more formulating agents or additives.
(18) The skin care preparation according to (16), wherein the skin care preparation is in the form of a gel, an emulsion, an oil-in-water or water-in-oil two-phase emulsion, a mask, a lotion, a concentrate, a serum, a nanocapsule, a liposome or a lipstick.
Compared with the prior art, the invention has the following advantages and positive effects:
the scutellarin and the analogue thereof can be combined with the vascular endothelial growth factor to increase the migration of skin keratinocytes induced by the vascular endothelial growth factor and the platelet growth factor group, thereby promoting the healing of wounds. The mechanism of the action is found in the invention that scutellarin and analogues thereof can promote phosphorylation of a vascular endothelial growth factor receptor 2(VEGFR2) of an upper epidermal cell by combining with a vascular endothelial growth factor, so that the scutellarin and the analogues thereof are activated, expression and phosphorylation of ErK in a downstream signal path MAPK of the receptor are increased, and expression of matrix metalloproteinase (MMP9) related to keratinocyte migration is increased.
Based on the findings, the invention provides a new application of scutellarin and analogues thereof in promoting wound healing and repair, which can coordinate and increase the effects of Vascular Endothelial Growth Factor (VEGF) and platelet growth factor group (SGC) in this respect, so that the scutellarin and analogues thereof can be used as medicines or cosmetics for treating and/or relieving skin wounds and/or scars, and have good clinical application prospect and market prospect.
In addition, the scutellarin is free of cytotoxicity and reliable in safety.
Drawings
FIG. 1 shows the result of cytotoxicity analysis of scutellarin.
Fig. 2 shows a photograph (a) of scutellarin promoting vascular endothelial growth factor-induced scratch healing of keratinocytes (HaCaT) and a result of quantifying wound healing rate (B).
Fig. 3 shows a photograph (a) of scutellarin-promoted platelet growth factor group-induced scratch healing of keratinocytes (HaCaT) and a result of quantifying wound healing rate (B).
FIG. 4 is a graph showing the results of RT-PCR quantification of the transcriptional level of scutellarin-promoted VEGF (A) or SGC (B) induced keratinocyte migration-associated gene matrix metalloproteinase-9 (MMP 9);
fig. 5 shows western blots (a and C) of scutellarin in combination with SGC to phosphorylate Erk in vegf-specific receptor 2(VEGFR2) and its downstream signaling pathway and corresponding quantification plots (B and D) of band intensities, where the ordinate of plots B and D is a multiple relative to baseline, and the band intensities of the blank control group are taken as baseline.
FIG. 6 shows a computer simulation of scutellarin (A), baicalin (B) and breviscapine (C) binding to vascular endothelial growth factor molecules.
Detailed Description
The present invention is further described in the following description of the embodiments with reference to the drawings, which are not intended to limit the invention, and those skilled in the art may make various modifications or improvements based on the basic idea of the invention, but within the scope of the invention, unless departing from the basic idea of the invention.
Scutellarin is also called as adonixin, and the plant source is stems and leaves of Scutellaria baicalensis Georgi of Labiatae and herba Scutellariae Barbatae whole plant, etc., and the structural formula is shown in formula I below.
Clinical research results show that scutellarin has the effects of reducing cerebrovascular resistance, improving cerebral blood circulation, increasing cerebral blood flow and resisting platelet aggregation, and is clinically used for treating paralysis caused by cerebrovascular diseases. However, the influence and action mechanism of scutellarin on skin wound healing and skin keratinocytes are not reported.
The scutellarin and the analogue thereof can be combined with the vascular endothelial growth factor, so that the migration of skin keratinocytes induced by the vascular endothelial growth factor and the platelet growth factor group is effectively increased, and the healing of wounds is promoted.
The invention also discovers that the mechanism of the action is that scutellarin and analogues thereof can effectively promote phosphorylation of a vascular endothelial growth factor receptor 2(VEGFR2) of an upper epidermal cell by combining with a vascular endothelial growth factor, so that the scutellarin and the analogues thereof are activated, and expression and phosphorylation of ErK in a downstream signal pathway MAPK of the receptor are increased, so that a MAPK signal pathway mediated by VEGFR2 can be activated, and the function of promoting migration of stratum corneum cells is exerted. In stratum corneum cells, VEGFR2 and its mediated signaling pathways are important for regulating cell survival, proliferation, and migration. Meanwhile, scutellarin and analogues thereof can effectively increase the expression of matrix metalloproteinase (MMP9) of genes related to keratinocyte migration promoted by VEGF or SGC.
Therefore, the scutellarin and the analogue thereof can enhance the biological function of the vascular endothelial growth factor and the platelet growth factor group for inducing wound healing, and can be used as a wound healing agent.
Based on the findings, the invention provides the application of scutellarin and analogues thereof in preparing medicines or skin care products for promoting the effects of vascular endothelial growth factor and/or platelet growth factor groups, wherein the analogues of the scutellarin are selected from one or more of baicalin and breviscapine.
The structural formula of baicalin is shown in formula II below, and the structural formula of breviscapine is shown in formula III below.
The effects of promoting vascular endothelial growth factor and/or platelet growth factor populations include promoting keratinocyte migration, promoting wound repair and/or healing, promoting matrix metalloproteinase expression, and activating VEGFR2 and its mediated MAPK signaling pathways.
Therefore, scutellarin and its analogs can promote keratinocyte migration, promote wound repair and/or healing, promote matrix metalloproteinase expression, activate VEGFR2, and activate VEGFR 2-mediated MAPK signaling pathway.
Therefore, the invention also provides application of scutellarin and analogues thereof in preparing medicines or skin care products for promoting migration of keratinocytes, promoting wound repair and/or healing, promoting expression of matrix metalloproteinase, activating VEGFR2, and/or activating VEGFR 2-mediated MAPK signaling pathway.
In the present invention, the term "promoting the action of vascular endothelial growth factor and/or platelet growth factor group" means that the scutellarin and the analogues thereof enhance the function as compared with the function exerted by vascular endothelial growth factor and platelet growth factor group alone, and that the vascular endothelial growth factor and platelet growth factor group includes the vascular endothelial growth factor and platelet growth factor group of the body itself and the vascular endothelial growth factor and platelet growth factor group used exogenously.
In the present invention, the term "platelet growth factor group" refers to platelet rich plasma extracted from whole blood, which includes vascular endothelial growth factor VEGF, epidermal growth factor EGF, platelet derived growth factor PDGF, platelet 4 factor PF4 and β -platelet globulin, and these components are dispersed in the platelet rich plasma. The population of platelet growth factors can be obtained by the method described in example 2.
In some embodiments, the present invention provides the use of scutellarin and analogs thereof in the preparation of a medicament for treating and/or alleviating skin wounds and/or scars.
In other embodiments, the present invention provides the use of scutellarin and analogs thereof in the preparation of a skin care product for treating and/or alleviating skin wounds and/or scars.
Based on the application of the scutellarin and the analogue thereof, the scutellarin and/or the analogue thereof can be independently prepared into medicines or prepared into skin care products, and after the application, the scutellarin and/or the analogue thereof can enhance the effects of VEGF and SGC in vivo; or used in combination with other VEGF and/or SGC drugs or products to enhance the effects of the VEGF and/or SGC drugs or products. Scutellarin and/or its analogs can also be combined with VEGF and/or SGC to make into medicine or cosmetic.
Therefore, the invention also provides a pharmaceutical composition, which is characterized by comprising the vascular endothelial growth factor and the scutellarin and/or the analogue thereof, wherein the analogue of the scutellarin is selected from one or more of baicalin and breviscapine.
Preferably, the molar ratio of the vascular endothelial growth factor to the scutellarin or the analogue thereof is 1 (4000-12000). In the case where the pharmaceutical composition comprises scutellarin and analogues thereof, the molar ratio of vascular endothelial growth factor to the sum of the two is preferably 1 (4000-12000).
In some embodiments, the pharmaceutical composition comprises a platelet growth factor group comprising vascular endothelial growth factor, in which case the molar ratio of vascular endothelial growth factor to said scutellarin or analog thereof in the platelet growth factor group can be 1 (30000-150000). In the case where the pharmaceutical composition comprises scutellarin and analogues thereof, the molar ratio of the vascular endothelial growth factor in the platelet growth factor group to the sum of the two may be 1 (30000-150000).
The invention also provides application of the pharmaceutical composition in preparing a medicament or a skin care product for promoting the effect of the vascular endothelial growth factor and/or the platelet growth factor group.
In some embodiments, the effects include promoting keratinocyte migration, promoting wound repair and/or healing, promoting expression of matrix metalloproteinases, activating vascular endothelial growth factor-specific receptor 2 and its mediated MAPK signaling pathway.
In some embodiments, the medicament is a medicament for treating and/or alleviating a skin wound and/or scar.
In other embodiments, the skin care product is a skin care product for treating and/or alleviating skin wounds and/or scars.
The invention also provides a medicament for promoting the effect of vascular endothelial growth factor and/or platelet growth factor groups, which comprises the pharmaceutical composition of the invention.
In some embodiments, the medicament is for treating and/or alleviating a skin wound and/or scar.
A single dose of the medicament may contain a daily dose of the active ingredient.
Where the medicament comprises one of scutellarin and its analogues, the daily dose of scutellarin may be 10-200 mg; the daily dosage of scutellarin analogue can be 10-200 mg. When the drug comprises both scutellarin and its analogs, the daily dose of the combination of scutellarin and its analogs may be 10-200 mg.
The daily dose of the vascular endothelial growth factor may be 10-30 μ g. When the drug comprises a population of platelet growth factors, the daily dose of the population of platelet growth factors may be 10-100 μ g.
According to practical applications, the above-mentioned pharmaceutical composition of the present invention may comprise scutellarin and/or its analogs, and vascular endothelial growth factor or platelet growth factor groups in amounts used to formulate a single dose of the drug, and may also comprise these active ingredients in amounts used to formulate multiple doses of the drug.
The medicament can be in the form of liniment or injection.
The medicament may also comprise pharmaceutically acceptable adjuvants.
The excipients may be selected according to the desired dosage form. For example, when the dosage form is liniment, the auxiliary material can be one or more selected from glycerol, allantoin, xanthan gum, squalane and phenoxyethanol; when the preparation form is intravenous injection, the auxiliary material can be one or more selected from mannitol, lactose, dextran, xylitol, sorbitol glucose and sodium chloride.
The content of the medicinal auxiliary materials in the single-dose medicament can be adjusted within the range commonly used in the field according to the actual requirement.
The invention also provides a skin care product for promoting the effect of the vascular endothelial growth factor and/or the platelet growth factor group, which comprises the pharmaceutical composition.
In some embodiments, the skin care products are used to treat and/or alleviate skin wounds and/or scars.
A single dose of the skin care product may contain a daily dose of the active ingredient.
When the skin care product contains one of scutellarin and analogues thereof, the daily dose of scutellarin can be 10-200 mg; the daily dosage of scutellarin analogue may be 10-200 mg. When the skin care product comprises both scutellarin and its analogs, the daily dose of the combination of scutellarin and its analogs may be 10-200 mg.
The daily dose of the vascular endothelial growth factor may be 10-30 μ g. When the skin care product comprises a platelet growth factor population, the daily dose of the platelet growth factor population may be 10-100 μ g.
The skin care product can be in the form of gel, lotion, oil-in-water or water-in-oil dual phase lotion, pack, lotion, concentrate, essence, nanocapsule, liposome, or lipstick.
The skin care product may also comprise one or more formulating agents or additives. In some embodiments, the formulating agent or additive is selected from the group consisting of penetrants, thickeners, surfactants, emollients, polydimethylsiloxanes, cyclomethicones, emulsifiers, preservatives, oils, UV-a and UV-B filters, pigments, dyes, film formers, minerals, and fragrances.
The content of the formulation or additive in the single-dose skin care product can be adjusted within the range commonly used in the field according to actual needs.
The present invention will be further explained or illustrated below by way of examples, which should not be construed as limiting the scope of the invention.
Examples of the present invention
Unless otherwise indicated, the experimental procedures used in the following examples were performed using conventional experimental protocols, procedures, materials and conditions known in the art.
Example 1: cytotoxicity assays
To evaluate whether scutellarin is cytotoxic in stratum corneum cells and stimulates skin cell proliferation, MTT assay was used to analyze cell viability.
Human HaCaT keratinocytes (purchased from adex bio, san diego) were cultured in DMEM medium supplemented with 10% FBS, 1% penicillin/streptomycin for 2 days to 80% growth density. Adherent cells at the bottom of the dish were digested with a 0.25% trypsin solution to prepare a cell suspension, which was counted and then prepared to have a cell concentration of 1X 104one/mL of the suspension and seeded in 96-well plates, 1000 cells per well, into CO2Incubator (5% CO)2Incubated at 37 ℃ for 24 hours. Then scutellarin with different final concentrations as shown in FIG. 2 was added for further culture for 24 hours, the medium in the 96-well plate was aspirated, 0.5mg/mL MTT solution was added, and the mixture was cultured in an incubator for 4 hours. After incubation, the cells were analyzed for viability by adding DMSO solution and shaking for 15 minutes on a shaker and measuring the absorbance at 570 nm.
As shown in FIG. 1, HaCaT keratinocytes did not show a significant reduction in cell number at concentrations of 0.5-100. mu.M scutellarin. The results indicate that scutellarin does not adversely affect skin cells, and the safety is reliable.
Example 2: keratinocyte scratch test
As shown in fig. 2A and B, scutellarin at 3 μ M alone did not promote healing of keratinocyte scratch, whereas experimental group 2 apparently healed at20 hours and at a significantly faster rate than the other groups, indicating that 3 μ M scutellarin enhanced VEGF-induced keratinocyte migration.
Since VEGF is highly enriched in the platelet growth factor population (SGC), the scratched HaCaT cell monolayers were drug-treated with the platelet growth factor population in the same way: cells treated with high concentration 0.5% (v/v) SGC alone were used as positive controls; cells treated with 0.5% (v/v) SGC and avastin (200. mu.g/mL) were used as negative controls; cells without any drug treatment (with an equal volume of DMEM added) were used as a blank; experimental group 1 was cells treated with 3. mu.M scutellarin and low concentration of 0.1% (v/v) SGC, experimental group 2 was cells treated with 3. mu.M scutellarin alone, and experimental group 3 was cells treated with low concentration of 0.1% (v/v) SGC alone.
As shown in fig. 3, the composition of scutellarin and platelet growth factor group significantly promoted migration of HaCaT cells in the scratch space compared to the control group, indicating that scutellarin can promote the growth factor or platelet growth factor group to induce keratinocyte migration.
The preparation method of the platelet growth factor group comprises the following steps: after collecting human whole blood with an anticoagulant, the whole blood sample was centrifuged at 3800rpm for 20 minutes at 4 ℃. Whole blood plasma was separated after the first centrifugation. The plasma was then centrifuged at 1250rpm for 10 minutes at 4 ℃. The supernatant of 2/3 was discarded as platelet poor plasma and the remainder 1/3 was resuspended, called Platelet Rich Plasma (PRP). A population of platelet growth factors was obtained by adding 10mM EDTA/EGTA protease inhibitor to PRP, removing ferrous ions using a magnetic support (from Merck Millipore, Burlington, Mass., USA), and exfoliating platelets by one freeze-thaw cycle (-80 ℃ for 10 hours and 37 ℃ for 10 minutes). After UV sterilization for about 30 minutes, the plasma samples were placed in Labconco
Freeze-dried in a freeze-drying system (Labconco, missouri) for 16 hours under vacuum and stored at 4 ℃ until use.
Example 3: the scutellarin in combination with VEGF or SGC can promote expression of matrix metalloproteinase-9 (MMP9) related to keratinocyte migration
To further confirm the effect of scutellarin in combination with VEGF or SGC on keratinocyte migration, the transcription level of MMP9 gene was measured using RT-PCR method. First, HaCaT cells were prepared to a cell concentration of 3X 105one/mL of the suspension and plated on 12-well plates, 3X 10 per well5And (4) cells. Put in CO2Incubator (5% CO)237 ℃) for 24 hours, then the drugs at different concentrations were added: the dosing setup was essentially the same as example 2, except that this example added the cell group treated with 1 μ M scutellarin and low concentration 0.1% (v/v) SGC (Experimental group 4), and the cell group treated with 3 μ M scutellarin and medium concentration 0.3% (v/v) SGC (Experimental group 5). After incubation for a further 24 hours after dosing, mRNA was extracted from HaCaT cells and RT-PCR was performed using primers for MMP 9. The primer sequences are as follows: MMP 9F-primer 5'-GGAGCGAGATCCCTCCAAAAT-3' (SEQ ID NO.1) and MMP 9R-primer 5'-GGCTGTTGTCATACTTCTCATGG-3' (SEQ ID NO. 2).
As shown in fig. 4, scutellarin significantly promoted VEGF or SGC-induced transcription of MMP9, which was enhanced by about 130% of that of the blank control. These results indicate that scutellarin in combination with VEGF or SGC has a significant effect on promoting wound healing by increasing the expression of MMP 9.
Example 4: scutellarin specifically activates vascular endothelial growth factor specific receptor 2(VEGFR2) and mediated signal path thereof, and plays a role in promoting wound healing
The effect of scutellarin on VEGF-dependent wound healing in platelet growth factor populations was evaluated using western blot experiments. HaCaT cells were seeded in 12-well plates (1.5X 10)5Cells/well) in CO2Incubator (5% CO)2Incubated at 37 ℃ for 24 hours. Adding serum-free DMEM, starving for 3 hr, and adding 0.5% (v/v) SGC, 0.1% (v/v) SGC, 3 μ M scutellarin, and a combination of 0.1% (v/v) SGC and 3 μ M scutellarin. Samples were collected for 0, 5, and 10 minutes, HaCaT cells were lysed using a low concentration lysate (100mM Tris-Cl pH6.8, 8% sodium dodecyl sulfate, 0.2% bromophenol blue, 20% glycerol, 400mM 2-mercaptoethanol), and cell lysates were collected and loaded on 8% SDS-PAGE gels. The antibodies used for western blotting were: 1:1000 Phospho-VEGFR2 rabbit anti-monoclonal antibody (CST,2478S),1:1000 VEGFR2 rabbit anti-monoclonal antibody (CST,2479S), 1:1000 Phospho-p44/42MAPK (Erk1/2) (CST,9101S),1:1000p44/42MAPK (Erk1/2) (CST, 8544S).
The results are shown in FIGS. 5A-D. FIGS. 5A and B show that scutellarin in combination with low concentration SGC induced phosphorylation of VEGFR2, with the levels of phosphorylated VEGFR2 in the blank control group (no drug added but with an equal volume of DMEM) and the low concentration SGC treated group not significantly changed after 5 minutes and 10 minutes, phosphorylated VEGFR2 in the high concentration SGC treated group about 3-fold after 10 minutes compared to about 0 minutes, and phosphorylated VEGFR2 in the combination of 0.1% (v/v) SGC and 3. mu.M scutellarin in the positive control group (0.5% (v/v) SGC alone) about 2.5-fold compared to the positive control. FIGS. 5C and D show the Erk phosphorylation downstream of this receptor induced by scutellarin in combination with low concentrations of SGC, where the level of phosphorylated Erk expression increased about 1.5-fold after 10 minutes in the 0.1% (v/v) SGC/3. mu.M scutellarin combination group, comparable to the effect induced by 0.5% (v/v) SGC in the positive control group.
Example 5: molecular fitting experiment
Molecular interaction between scutellarin and its analogs and vascular endothelial growth factor was investigated using Molecular packing (Vina, PhyMOL). The molecular docking was simulated using PyMOL molecular image analysis software and SwissDock tool to find scutellarin and its analogues: breviscapine and baicalin can bind with VEGF. Binding affinity data were obtained from the molecular docking tool SwissDock, detailed procedure as follows:
1. inputting the pdb structure file of target protein VEGF, the PDb number of VEGF used in the invention is 3 QTK.
2. Preparing a small molecule structure mol2 file of small molecules (scutellarin, breviscapine and baicalin).
3. The pdb and mol2 files were uploaded on the SwissDock website, and the molecular docking scores were obtained by clicking on the submission. As shown in the following table:
the structural interaction between scutellarin and vascular endothelial growth factor is shown in figure 6A. The binding affinity of scutellarin and VEGF protein is between-5.57 and-8.28, which indicates that the binding affinity of scutellarin and VEGF protein is stronger. The structural interaction of scutellarin, breviscapine and VEGF of the scutellarin analogues is shown in FIGS. 6B and C, the binding affinity is-5.59-8.15 and-6.34-8.26 respectively, which shows that the binding affinity of the scutellarin analogues and the breviscapine is also stronger.
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Claims (18)
1. A pharmaceutical composition, comprising vascular endothelial growth factor and scutellarin and/or analogues thereof, wherein the analogues are selected from one or more of baicalin and breviscapine.
2. The pharmaceutical composition of claim 1, wherein the molar ratio of the VEGF to the scutellarin or the analog thereof is 1 (4000-12000).
3. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition comprises a population of platelet growth factors.
4. The pharmaceutical composition according to claim 3, wherein the molar ratio of the vascular endothelial growth factor and the scutellarin or the analogues thereof in the platelet growth factor group is 1 (30000-150000).
5. Application of scutellarin and analogues thereof in preparing medicines or skin care products for promoting effects of vascular endothelial growth factor and/or platelet growth factor groups, wherein the analogues are selected from one or more of baicalin and breviscapine.
6. The use according to claim 5, wherein the effects include promoting keratinocyte migration, promoting wound repair and/or healing, promoting expression of matrix metalloproteinases, activating vascular endothelial growth factor-specific receptor 2 and its mediated MAPK signaling pathway.
7. Use according to claim 5, wherein the medicament is a medicament for the treatment and/or alleviation of skin wounds and/or scars.
8. Use according to claim 5, wherein the skin care product is a skin care product for the treatment and/or alleviation of skin wounds and/or scars.
9. Use of a pharmaceutical composition according to any one of claims 1 to 4 for the preparation of a medicament or a skin care product for promoting the action of vascular endothelial growth factor and/or platelet growth factor groups.
10. The use according to claim 9, wherein the effects include promoting keratinocyte migration, promoting wound repair and/or healing, promoting expression of matrix metalloproteinases, activating vascular endothelial growth factor-specific receptor 2 and its mediated MAPK signaling pathway.
11. Use according to claim 9, wherein the medicament is a medicament for the treatment and/or alleviation of skin wounds and/or scars.
12. Use according to claim 9, wherein the skin care product is a skin care product for the treatment and/or alleviation of skin wounds and/or scars.
13. A medicament for promoting the action of a population of vascular endothelial growth factor and/or platelet growth factor comprising the pharmaceutical composition of any one of claims 1-4.
14. The medicament of claim 13, wherein the medicament further comprises a pharmaceutically acceptable excipient.
15. The medicament of claim 13, wherein the medicament is in the form of a liniment or an injection.
16. A skin care product for promoting the action of a group of vascular endothelial growth factor and/or platelet growth factor comprising the pharmaceutical composition according to any one of claims 1 to 4.
17. The skin care product of claim 16, wherein the skin care product further comprises one or more formulating agents or additives.
18. The skin care product of claim 16, wherein the skin care product is in the form of a gel, an emulsion, an oil-in-water or water-in-oil dual phase emulsion, a mask, a lotion, a concentrate, a serum, a nanocapsule, a liposome, or a lipstick.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115887528A (en) * | 2022-11-23 | 2023-04-04 | 广东药科大学 | Biological rehabilitation preparation for promoting stem cell function and further repairing osteoarthritis |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102247397A (en) * | 2011-05-30 | 2011-11-23 | 浙江大学 | Application of astragaloside to preparation of drug |
| CN102349925A (en) * | 2011-08-23 | 2012-02-15 | 上海中医药大学附属岳阳中西医结合医院 | Application of astragaloside in preparation of drugs for promoting epithelization in wound healing |
| CN103655547A (en) * | 2013-12-18 | 2014-03-26 | 成都中医药大学 | Novel application of scutellarein |
| CN104739913A (en) * | 2015-03-26 | 2015-07-01 | 鼎旺生物科技有限公司 | Preparation for accelerating wound healing, and preparation method and application thereof |
-
2020
- 2020-10-30 CN CN202011191253.3A patent/CN114432430B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102247397A (en) * | 2011-05-30 | 2011-11-23 | 浙江大学 | Application of astragaloside to preparation of drug |
| CN102349925A (en) * | 2011-08-23 | 2012-02-15 | 上海中医药大学附属岳阳中西医结合医院 | Application of astragaloside in preparation of drugs for promoting epithelization in wound healing |
| CN103655547A (en) * | 2013-12-18 | 2014-03-26 | 成都中医药大学 | Novel application of scutellarein |
| CN104739913A (en) * | 2015-03-26 | 2015-07-01 | 鼎旺生物科技有限公司 | Preparation for accelerating wound healing, and preparation method and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| 丁璐等: "富血小板血浆治疗老年慢性难愈合伤口的疗效", 《中国老年学杂志》 * |
| 高钟秀子: "野黄芩苷促进体外内皮细胞的血管生成", 《中国优秀硕士学位论文全文数据库》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115887528A (en) * | 2022-11-23 | 2023-04-04 | 广东药科大学 | Biological rehabilitation preparation for promoting stem cell function and further repairing osteoarthritis |
| CN115887528B (en) * | 2022-11-23 | 2023-12-12 | 广东药科大学 | Biological rehabilitation preparation for promoting stem cell function and further repairing osteoarthritis |
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| CN114432430B (en) | 2024-01-26 |
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