CN115607606B

CN115607606B - Application of capsicum-derived nano vesicles in preparation of drugs for preventing and treating atherosclerosis diseases

Info

Publication number: CN115607606B
Application number: CN202211629284.1A
Authority: CN
Inventors: 王小庆; 柯晓; 杨荣丰; 陈俊羽; 彭长农
Original assignee: Fuwai Hospital of CAMS and PUMC
Current assignee: Fuwai Hospital of CAMS and PUMC
Priority date: 2022-12-19
Filing date: 2022-12-19
Publication date: 2023-03-17
Anticipated expiration: 2042-12-19
Also published as: CN115607606A

Abstract

The invention belongs to the technical field of biological medicines, and discloses application of nano vesicles derived from hot pepper in preparation of medicines for preventing and treating atherosclerotic diseases. The nano vesicle derived from the capsicum is obtained by extracting and separating the capsicum with water, has natural components, no toxic or side effect and good biocompatibility and safety, and thus has wide application prospect. In vivo and in vitro experiments are carried out by utilizing the nano vesicles from the hot pepper, the nano vesicles from the hot pepper are found to have strong effects of protecting endothelial cells and resisting atherosclerosis, can be orally absorbed, have no tissue and vascular irritation, and provide important basis for the application of the nano vesicles from the hot pepper in preventing and treating atherosclerosis.

Description

Application of capsicum-derived nano-vesicles in preparation of medicines for preventing and treating atherosclerosis diseases

Technical Field

The invention belongs to the technical field of biological medicines, and particularly relates to application of capsicum-derived nano vesicles in preparation of medicines for preventing and treating atherosclerotic diseases.

Background

Atherosclerosis (AS) is characterized by the deposition of lipids and other substances within the arterial wall, forming AS plaques, closely associated with an increased risk of various life-threatening cardiovascular and cerebrovascular diseases, such AS stroke and myocardial infarction. Endothelial function and structural integrity are essential for maintaining vascular homeostasis, and dysfunction of vascular endothelial cells is considered to be an important factor in the pathogenesis and progression of AS. Oxidized low-density lipoprotein (oxLDL) is a major risk factor for AS.

Ox-LDL may regulate the development of AS through a variety of mechanisms, including the induction of endothelial cell damage and dysfunction. The research of preventing and treating AS by reducing vascular endothelial injury is always a hotspot of the research of cardiovascular and cerebrovascular diseases, and targeted oxLDL resistance is a promising therapeutic target for the injury of endothelial cells. Studies have reported that regular capsicum ingestion can achieve significant cardiovascular benefits, but the mechanism is unknown. Capsaicin (CAP), one of the most abundant and approved active ingredients in pepper, induces the activation of endothelial nitric oxide synthase (eNOS) of endothelial cells, protects endothelial cells from leukocyte adhesion, and inhibits ROS production and caspase-3 activation of oxLDL pretreated endothelial cells. However, oral bioavailability of capsaicin is extremely low, and effective endothelial protection is difficult to obtain by oral capsaicin. It is therefore believed that the protective effect of capsicum in vivo may not be derived solely from free capsaicin. Meanwhile, intravenous capsaicin causes severe local tissue inflammation and phlebitis, which is another important factor that clinical application of capsaicin is limited.

Disclosure of Invention

In order to overcome the defects of the prior art, the invention mainly aims to provide the application of Pepper Derived Nano Vesicles (PDNV) in preparing the medicines for preventing and treating the atherosclerosis.

The purpose of the invention is realized by the following scheme:

application of pepper-derived nano vesicles (PDNV) in preparation of medicines for preventing and treating atherosclerosis diseases.

Application of pepper-derived nano vesicles (PDNV) in preparation of medicines for preventing atherosclerosis diseases.

Application of pepper-derived nano vesicles (PDNV) in preparation of medicines for treating atherosclerosis diseases.

The medicines are the same or different and respectively comprise a therapeutically effective amount of the nano vesicles derived from the capsicum.

The medicines which are the same or different can be respectively prepared into various medicinal dosage forms by adopting a conventional method, and the dosage forms comprise: tablet (sugar-coated tablet, film-coated tablet, enteric-coated tablet, etc.), capsule (hard capsule, soft capsule, etc.), oral liquid, buccal agent, granule, pill, pellet, suspension, medicated wine, tincture, drop, etc., and injection, etc., for oral administration and other dosage forms, such as injection, etc.

The medicines which are the same or different respectively can also contain one or more than one pharmaceutically acceptable carriers or excipients.

The excipients may include diluents, wetting agents, lubricants, fillers, preservatives and the like.

The atherosclerotic disease can include atherosclerosis or coronary heart disease caused by atherosclerosis, cerebral infarction, cerebral apoplexy, myocardial infarction, arrhythmia or peripheral vascular disease.

The nano vesicle derived from the pepper can be separated from the pepper.

The capsicum-derived nano-vesicles can be obtained by separating through a conventional separation method. For example, the capsicum can be obtained by pulverizing or breaking cell wall and extracting with extractant.

Further, the extract obtained by extraction may be subjected to high-speed centrifugation.

Furthermore, the extract can be pre-centrifuged to remove precipitates, and then high-speed centrifugation is performed to obtain the precipitates, namely the pepper-derived nano vesicles.

The speed of the pre-centrifugation may be 10000g or less. Non-target substances can be separated and removed through pre-centrifugation, and the influence of impurities on subsequent centrifugal separation is reduced.

The speed of the ultracentrifugation may be 100000g or more.

The extractant may be water or an aqueous solution added with a buffer such as phosphate.

The obtained pepper-derived nano vesicles can be resuspended by using buffer solutions such as PBS and the like so as to obtain a pepper-derived nano vesicle suspension.

The pepper-derived nanovesicle suspension may be filtered using a filter before use to obtain a sterile pepper-derived nanovesicle suspension. The filter may be a 0.22 μm filter.

Specifically, the invention also provides a capsicum-derived nano vesicle which is separated from capsicum; the preparation method further comprises the following steps: crushing fresh Capsici fructus, soaking in water, separating to obtain supernatant, pre-centrifuging the supernatant under centrifugal force of 100-10000g for several times, such as 300g, 2000g, and 10000g for 10-30min, respectively collecting supernatant, and discarding precipitate; ultracentrifugation is carried out on the supernatant after the pre-centrifugation at a centrifugal force of 100000g or more (such as 135000 g) for 10-100min, and the obtained precipitate is the nano vesicle derived from the capsicum.

Furthermore, the pepper can be cleaned and dried before use.

Further, the obtained pepper-derived nano vesicles are re-suspended by utilizing PBS (phosphate buffer solution) to obtain a pepper-derived nano vesicle suspension.

Further, the suspension of pepper-derived nanovesicles can be further purified by ultracentrifugation at a centrifugal force of 100000g or more (e.g., 135000 g).

The nano vesicle derived from the capsicum is obtained by extracting and separating the capsicum with water, has natural components, no toxic or side effect and good biocompatibility and safety, and thus has wide application prospect.

The invention utilizes the nano vesicles from the capsicum to carry out in vivo and in vitro experiments, and finds that the nano vesicles from the capsicum have strong effects of protecting endothelial cells and resisting atherosclerosis, can be orally absorbed, have no tissue and vascular irritation, and provide important basis for the application of the nano vesicles from the capsicum in the prevention and treatment of the atherosclerosis.

Drawings

In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required in the embodiments will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and those skilled in the art can also obtain other related drawings based on the drawings without inventive efforts.

FIG. 1 is a transmission electron micrograph of PDNV.

FIG. 2 is a fluorescence plot (40X) of HUVEC uptake of DIL-PDNV.

FIG. 3 is a graph showing the effect of PDNV and CAP on HUVEC proliferation activity before and after ox-LDL intervention.

FIG. 4 is a graph showing the effect of PDNV on ox-LDL's ability to inhibit HUVEC migration.

FIG. 5 is a graph of the effect of PDNV on the ability of ox-LDL to inhibit the tube formation of HUVEC.

FIG. 6 is a graph of PDNV improving the level of inflammation in high fat fed Apoe-/-mice.

FIG. 7 is a graph of PDNV improving atherosclerosis in high fat fed Apoe-/-mice.

Wherein, the data in the figure are expressed as means + -SD, and P is less than 0.05; p is less than 0.01; p is less than 0.001.

Detailed Description

The present invention will be described in further detail with reference to examples, but the embodiments of the present invention are not limited thereto. The materials referred to in the following examples are commercially available without specific reference. The method is a conventional method unless otherwise specified.

In one embodiment, the application of the capsicum-derived nanovesicle (PDNV) in preparing the medicine for preventing and treating the atherosclerosis disease is provided.

In another embodiment, the application of the capsicum-derived nanovesicle (PDNV) in preparing a medicament for preventing atherosclerotic diseases is provided.

In another embodiment, the application of the pepper-derived nano-vesicle (PDNV) in preparing a medicament for treating the atherosclerotic disease is provided.

In one embodiment, the drugs are the same or different and respectively comprise a therapeutically effective amount of pepper-derived nanovesicles.

In one embodiment, the drugs are made into various pharmaceutical dosage forms by the same or different conventional methods, and the dosage forms comprise: tablet, sugar-coated tablet, film-coated tablet, enteric-coated tablet, capsule, hard capsule, soft capsule, oral liquid, buccal agent, granule, pill, pellet, suspension, medicated wine, tincture, drop, etc., and injection, etc., such as injection.

In one embodiment, the drugs, which may be the same or different, each further comprise one or more pharmaceutically acceptable carriers or excipients.

In one embodiment, the excipients include diluents, wetting agents, lubricants, fillers, preservatives, and the like.

In one embodiment, the atherosclerotic disease includes atherosclerosis or coronary heart disease caused by atherosclerosis, cerebral infarction, cerebral stroke, myocardial infarction, arrhythmia or peripheral vascular disease.

In one embodiment, the pepper-derived nanovesicles are isolated from pepper.

In one embodiment, the pepper-derived nanovesicles are isolated by a conventional isolation method. In one embodiment, the capsicum is obtained by crushing or breaking the wall of capsicum and adding an extracting agent for extraction and separation.

In one embodiment, the extract obtained by extraction is subjected to high-speed centrifugation.

In one embodiment, the extract is pre-centrifuged to remove the precipitate, and then high-speed centrifugation is performed to obtain the precipitate, i.e., the pepper-derived nano vesicles.

In one embodiment, the speed of the pre-centrifugation is 10000g or less. Non-target substances can be separated and removed through pre-centrifugation, and the influence of impurities on subsequent centrifugal separation is reduced.

In one embodiment, the speed of the ultracentrifugation is 100000g or more.

In one embodiment, the extractant is water or an aqueous solution added with a buffer such as phosphate.

In one embodiment, the obtained pepper-derived nanovesicles are resuspended in a buffer such as PBS to obtain a pepper-derived nanovesicle suspension.

In one embodiment, the capsicum-derived nanovesicle suspension is filtered by a filter before use to obtain a sterile capsicum-derived nanovesicle suspension.

In one embodiment, the filter is a 0.22 μm filter.

In one embodiment, a capsicum-derived nanovesicle is isolated from capsicum; the preparation method specifically comprises the following steps: crushing fresh Capsici fructus, soaking in water, separating to obtain supernatant, pre-centrifuging the supernatant under centrifugal force of 100-10000g for several times, such as 300g, 2000g, and 10000g for 10-30min, respectively collecting supernatant, and discarding precipitate; ultracentrifugation is carried out on the supernatant after the pre-centrifugation at or above 100000g (such as 135000 g) of centrifugal force for 10-100min, and the obtained precipitate is the nano vesicle derived from the pepper.

In one embodiment, the peppers are cleaned and dried before use.

In one embodiment, the obtained pepper-derived nanovesicles are resuspended with PBS to obtain a pepper-derived nanovesicle suspension.

In one embodiment, the suspension of pepper-derived nanovesicles is further purified by ultracentrifugation at a centrifugal force of 100000g or more (e.g., 135000 g).

In one embodiment, PDNV is morphologically characterized by transmission electron microscopy. As observed by transmission electron microscopy (Tecnai G2 Spiritwin + GATAN 832.10W), PDNV is in the form of quasi-circular or oval vesicles (see FIG. 1), such as trays, with diameters ranging from 50 to 200nm.

In one example, the particle size and particle count of PDNV in a PDNV suspension is determined using a particle tracking analyzer (NanoSight NS 300). The results show that the PDNV mean diameter is 176.4 + -9.4 nm and the particle concentration is 1.0X 10 ¹¹ one/mL.

Example 1: UHPLC-MS/MS (ultra high Performance liquid chromatography-Mass Spectrometry/Mass Spectrometry) detection of Capsaicin (CAP) content in PDNV (PDNV)

(1) Preparation of samples: taking 20 mu L of PDNV suspension, adding 980 mu L of methanol, mixing uniformly, and standing for 1h at the temperature of minus 20 ℃; centrifuging at 12000rpm for 15min at 4 deg.C, and collecting supernatant for LC-MS/MS analysis;

(2) UHPLC-MS/MS detection: the target compound was chromatographed on a Waters TSS T3 column (2.1 mm × 100mm, 1.7 μm, agilent Technologies) using Agilent 1290 Infinity II series (Agilent Technologies) ultra performance liquid chromatograph. The liquid chromatography phase A is an aqueous solution containing 5mM ammonium formate and 0.1% formic acid, and the phase B is methanol. The flow rate of the mobile phase is 300 mu L/min, the temperature of the column incubator is 35 ℃, the temperature of the sample tray is 10 ℃, and the sample injection volume is 1 mu L. Standard solutions of the target compounds were introduced into the mass spectra before UHPLC-MS/MS analysis. In this test, all mass spectrometric data acquisition and target compound quantification Work was done by Agilent Mass Hunter Work Station Software (B.10.00, agilent Technologies). Preparing a series of calibration solutions by using a Capsaicin (CAP) standard substance, and carrying out UPLC-MRM-MS/MS analysis on the solutions to obtain a standard curve;

(3) The detection result showed that the concentration of CAP contained in PDNV was 340. Mu. Mol/L.

Example 2: ingestion of PDNV by HUVEC

Human Umbilical Vein Endothelial Cells (HUVEC) were purchased from qian new boat biotechnology limited, shanghai (zqxzbio, ZQ 0446);

(1) DIL-PDNV configuration: 10mg of DIL fluorescent dye probe (Solarbio, D8700) was mixed with 10 ¹¹ Co-incubating PDNV with a plurality of particles, and incubating for 30min at 37 ℃ in a dark place to obtain DIL-PDNV; ultracentrifugation is carried out at temperature of 4 deg.C of 100000g or more, such as centrifugation at 135000g for 70min, and precipitate is taken and resuspended in PBS buffer solution to obtain DIL-PDNV;

(2) P6 generation HUVEC were seeded on a confocal dish at a cell count of 1X 10 ⁵ After the cells are attached, the number of cells is 1X 10 ⁸ Adding DIL-PDNV at the concentration of one/mL, and incubating for 12h at 37 ℃ in a dark place;

(3) Changing the solution, washing with PBS for 2-3 times, fixing with paraformaldehyde for 30min, washing with PBS for 3 times after fixing, adding DAPI for nuclear staining for 30min;

(4) After staining, observation was performed using a fluorescence microscope, and the results are shown in FIG. 2. As can be seen, DAPI nuclear staining appears as blue staining, DIL-PDNV aggregates in the cytoplasm, and appears as a punctate red fluorescent signal.

Example 3: effect of PDNV on proliferative Activity of ox-LDL in intervention HUVEC

(1) The P6 generation HUVEC were inoculated into 96-well plates, 5000 cells were inoculated per well, and after the cells were fully adherent, PDNV (0,1X 10) containing different concentrations was replaced ⁷ 1X 10 units/mL ⁸ 1X 10 units/mL ⁹ 1X 10 units/mL ¹⁰ counts/mL) of ECM complete medium for 6h; CCK-8 reagent is added into each well according to the volume ratio of 10 to 1, the wells are incubated for 2 hours at 37 ℃, and an ultraviolet spectrophotometer detects the absorbance at 450nm, and the result is shown in figure 3. As can be seen in FIG. 3A, there is a concentration-dependent promotion of the proliferation activity of HUVEC cells by low and medium concentrations of PDNV; and inhibit its proliferation at high concentrations;

(2) And (3) intervention grouping: (1) NC group (blank control group), (2) CAP group (0.34. Mu. Mol/L), (3) PDNV group (1X 10) ⁸ one/mL); (according to the above, the PDNV concentration is 1X 10 ¹¹ At one/mL, the concentration of the included CAP is 340. Mu. Mol/L, where 0.34. Mu. Mol/L CAP and 1X 10 are selected ⁸ Equal concentrations of CAP contained in PDNV per mL);

inoculating HUVEC of P6 generation into a 96-well plate, inoculating 5000 cells into each well, replacing an ECM complete culture medium after the cells are completely attached to the wall, and respectively adding reagents according to grouping settings for intervention treatment for 6h; adding CCK-8 reagent into each well according to the volume ratio of 10 to 1, incubating for 2h at 37 ℃, and detecting the absorbance at 450nm by using an ultraviolet spectrophotometer, wherein the result is shown in figure 3. As can be seen in fig. 3B, PDNV has a significantly stronger effect on cell proliferation than CAP;

(3) And (3) intervention grouping: (1) NC group (blank control group), (2) ox-LDL group (100. Mu.g/mL), (3) PDNV (1X 10) ⁸ Per mL) + ox-LDL (100. Mu.g/mL), (4) CAP (0.34. Mu. Mol/L) + ox-LDL (100. Mu.g/mL);

inoculating the P6 generation HUVEC into a 96-well plate, inoculating 5000 cells into each well, and after the cells are completely attached to the wall, respectively adding reagents according to groups and then continuing to culture; in which group (3) is added PDNV (1X 10) ⁸ cell/mL) intervention is carried out for 6h, the solution is changed into a complete ECM culture medium, ox-LDL is added, and the culture is continued for 24h; group (4) adding CAP dryPre-culturing for 6h, changing the culture solution into complete ECM culture medium, adding ox-LDL, and continuously culturing for 24h; group (1) was incubated for 24 hours without adding any other reagent and group (2) with ox-LDL. After the culture is finished, adding the CCK-8 reagent into each well according to the volume ratio of 10. As can be seen in FIG. 3C, ox-LDL significantly inhibited the proliferative activity of HUVEC, and PDNV significantly reversed the proliferation-inhibiting effect of ox-LDL.

Example 4: effect of PDNV on ox-LDL inhibition of HUVEC migration

(1) And (3) intervention grouping: (1) NC group (blank control group), (2) PDNV group (1X 10) ⁸ Per mL), (3) ox-LDL group (100. Mu.g/mL), (4) PDNV (1X 10) ⁸ Per mL) + ox-LDL (100. Mu.g/mL), (5) CAP (0.34. Mu. Mol/L) + ox-LDL (100. Mu.g/mL);

(2) Inoculating the P6 generation HUVEC into a 12-pore plate, adding the reagents according to groups respectively after the cell fusion degree reaches 70%, and continuing to culture; in which the (4) th group is added with PDNV (1 × 10) ⁸ seed/mL) intervention is carried out for 6h, the liquid is changed into a complete ECM culture medium, ox-LDL is added, and culture is continued for 24h; adding CAP for intervention for 6h, changing the culture medium to complete ECM culture medium, adding ox-LDL, and culturing for 24 hr; the group (1) is not added with any other reagent, the group (2) is added with PDNV, the group (3) is added with ox-LDL, and the mixture is cultured for 24h respectively;

(3) After the culture is finished, adding pancreatin for digestion and heavy suspension, centrifuging to remove pancreatin, adding a serum-free culture medium, and counting cells;

(4) 1X 10 additions per transwell ⁴ The total volume of the corresponding number of cells is controlled to be 80-100 mu L; adding 400-450 mu L of ECM complete culture medium into the lower hole of the transwell chamber, and continuing to culture for 24h;

(5) PBS wash 1-2 times, methanol fixation for 20min, abandoning, staining with 1% crystal violet for 10min, washing with PBS, and taking pictures on the machine, wherein the blue stained cells are successfully migrated HUVEC, and the results are shown in FIG. 4. As can be seen, ox-LDL inhibited the migratory capacity of HUVEC, while PDNV significantly reversed this effect.

Example 5: effect of PDNV on ox-LDL inhibition of HUVEC in vitro tube formation

(2) Inoculating the P6 generation HUVEC into a 12-pore plate, adding the reagents according to groups respectively after the cell fusion degree reaches 70%, and continuing to culture; in which the group (4) is added with PDNV (1X 10) ⁸ cell/mL) intervention is carried out for 6h, the solution is changed into a complete ECM culture medium, ox-LDL is added, and the culture is continued for 24h; adding CAP for intervention for 6h, changing the culture medium to complete ECM culture medium, adding ox-LDL, and culturing for 24 hr; the group (1) is not added with any other reagent, the group (2) is added with PDNV, the group (3) is added with ox-LDL, and the culture is carried out for 24h respectively;

(3) After the culture is finished, sucking matrigel by using a precooled gun head, loading onto a precooled 96-well plate, spreading the matrigel on the bottom of each well by 80 mu L, and putting the plate into an incubator for incubation for 30-60min;

(4) After the incubation is completed, 1 ten thousand cells are added into each well according to groups, 3 multiple wells are arranged, the incubator is continuously cultured for 6 hours at 37 ℃ for photographing, and photographing is carried out under an inverted microscope, and the result is shown in fig. 5. As can be seen, ox-LDL significantly inhibited the tube-forming ability of HUVEC, while PDNV could significantly reverse this effect with significantly superior effect compared to CAP.

Example 6: PDNV improves atherosclerosis in high fat fed Apoe-/-mice

(1) Subject: 8 weeks old, male Apoe-/-mice;

(2) Experimental grouping: (1) control (PBS lavage); (2) stomach irrigation of PDNV (dose: 1X 10) ¹⁰ One/kg/time); (3) CAP was gavaged (dose: 0.034. Mu. Mol/kg/time) with 5 animals per group given 1 time every 3 days;

(3) The experimental steps are as follows: high fat feeding (21% milk fat, 0.15% cholesterol) was performed for 12 weeks with 3 gavages per week, depending on the group. After 12 weeks, the mice were euthanized and the blood, aorta, etc. tissues were left for further experiments;

(4) Measurement of inflammatory factors IL-1. Beta., IL-6, and IL-10: measurement of triglyceride, total cholesterol and low-density lipoprotein cholesterol in mouse serum was carried out according to the instructions of IL-1. Beta. Detection kit (Boxbio, AKFA 003C), IL-6 measurement kit (Jianheng, A111-1-1) and IL-10 measurement kit (A113-1-1), and the results are shown in FIG. 6. As can be seen, compared with the control group and the CAP group, the PDNV gastric perfusion group can reduce IL-1 beta and IL-6 (proinflammatory factors) in peripheral blood of mice and up-regulate the level of IL-10 (anti-inflammatory factors), the PDNV has good anti-inflammatory effect, but the CAP does not have the effect by oral administration;

(5) Aortal cryosection oil red O staining: taking out the autonomous arch of the mouse to the renal artery branch of the abdominal aorta, carrying out OCT embedding, manufacturing a frozen section, and then carrying out oil red O staining; the fatty plaque appeared orange-red when stained with oil red O, and the result is shown in fig. 7, where the blood vessel intima was red-stained and the clumps were atherosclerotic plaques. As can be seen, compared with the control group, the PDNV intragastric group can significantly reduce the area of aortic plaque, and the effect is significantly better than that of the CAP intragastric group.

The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such modifications are intended to be included in the scope of the present invention.

Claims

1. The application of the capsicum-derived nano vesicle in the preparation of drugs for preventing and treating atherosclerosis diseases;

the nano vesicles derived from the capsicum are obtained by separating the capsicum;

the capsicum-derived nano-vesicles are obtained by crushing or breaking walls of capsicum and extracting and separating with an extracting agent; pre-centrifuging the extracting solution, removing the precipitate, and then centrifuging at a high speed to obtain the precipitate, namely the pepper-derived nano vesicles;

the speed of the pre-centrifugation is 10000g or less;

the high-speed centrifugation speed is 100000g or more;

the extractant is water or water solution added with buffer solution.

2. Use according to claim 1, characterized in that: the medicine is prepared into various medicinal dosage forms, and the dosage forms comprise: tablet, capsule, oral liquid, granule, pill, suspension, medicated wine, tincture, and injection.

3. Use according to claim 1, characterized in that: the medicament also contains one or more pharmaceutically acceptable excipients.

4. Use according to claim 3, characterized in that: the excipient comprises at least one of diluent, wetting agent, lubricant, filler and preservative.

5. Use according to claim 1, characterized in that: the medicine also contains one or more than one pharmaceutically acceptable carriers.