阅读说明：本技术 左氧氟沙星在制备抗动脉粥样硬化药物中的应用 (Application of levofloxacin in preparation of anti-atherosclerosis drugs ) 是由 陈勇 容彤 盛青华 于 2021-06-15 设计创作，主要内容包括：本发明公开了左氧氟沙星在制备抗动脉粥样硬化药物中的应用,属于心血管疾病药效学的技术领域。本发明研究发现,HPβCD-Lev、Lev对高脂饮食造模动脉粥样硬化的影响,明确HPβCD-Lev抗动脉粥样硬化的药效。本发明将Lev、HPβCD-Lev用于心血管疾病特别是动脉粥样硬化疾病的延缓或治疗,扩展左氧氟沙星用药范围,降低疾病治疗成本,具有重要的研究价值和临床应用前景。(The invention discloses an application of levofloxacin in preparing an anti-atherosclerosis medicament, belonging to the technical field of cardiovascular disease pharmacodynamics. The research of the invention finds that the HP beta CD-Lev and the HP beta CD-Lev have influence on high-fat diet modeling atherosclerosis, and the drug effect of the HP beta CD-Lev on atherosclerosis resistance is determined. The invention uses Lev and HP beta CD-Lev for delaying or treating cardiovascular diseases, particularly atherosclerosis diseases, expands the medication range of levofloxacin, reduces the cost of disease treatment, and has important research value and clinical application prospect.)

1. The application of levofloxacin in preparing antiatherosclerotic medicine is characterized in that: the effective component of the medicine is levofloxacin.

2. Use according to claim 1, characterized in that: the drug is encapsulated with levofloxacin through hydroxypropyl-beta-cyclodextrin.

3. Use according to claim 1, characterized in that: the dosage of the levofloxacin is 10-80 mg/kg/2 d.

Technical Field

The invention belongs to the technical field of cardiovascular disease pharmacodynamics, and particularly relates to application of levofloxacin in preparation of an anti-atherosclerosis drug.

Background

In recent years, the number of Arteriosclerosis (arterosclerosis) has increased in China, which has become one of the main causes of death of the elderly and is still on the rise. Atherosclerosis (Atherosclerosis) is the most important one of the most common diseases in Atherosclerosis, and its formation and development is a slow, long-term process involving a variety of serum lipoproteins, a variety of cells (including vascular endothelial cells, macrophages, smooth muscle cells, lymphocytes, etc.), and a variety of molecules. Atherosclerosis can lead to a variety of diseases or clinical symptoms, the most well known and important of which are myocardial infarction, coronary heart disease, stroke, and the like. It can be said that atherosclerosis is the root cause of many important common diseases or deaths. Therefore, it is of great significance to find effective methods and drugs for preventing, diagnosing, delaying or treating atherosclerosis.

Hydroxypropyl-beta-cyclodextrin (HP beta CD) is prepared by substituting the 2-position hydrogen atom of hydroxyl on a beta cyclodextrin glucose residue by hydroxypropyl, a single molecule of the HP beta CD has a three-dimensional annular hollow structure with outer hydrophilicity and inner hydrophobicity like beta cyclodextrin, the HP beta CD is widely applied to various fields of food, medicine, industry and the like, the solubility, the availability, the stability and the like of the inclusion compound are increased by being used as a shell, and the application rate of the inclusion compound is greatly improved, for example, the solubility and the dissolution rate of itraconazole are obviously improved by using HP beta CD to include the itraconazole, the solubility of ibuprofen which is about 700 times increased by using HP beta CD to include an insoluble drug, and the solubilization is 70 times higher than that of the beta CD.

Levofloxacin (Levofloxacin, Lev) is a quinolone antibacterial drug having a resisting effect on many microorganisms, pathogenic factors or diseases, is commonly used for example for bacterial infections of urogenital systems, respiratory tract bacterial infections, gastrointestinal tract bacterial infections, skin soft tissue infections and the like, and achieves the effects of killing bacteria and resisting diseases mainly by inhibiting the activity of bacterial DNA helicase so that the bacteria cannot normally synthesize and replicate DNA and are finally self-killed. The invention uses levofloxacin in the delay treatment of atherosclerosis for the first time and achieves obvious drug effect.

Disclosure of Invention

Aiming at the defects and problems in the prior art, the invention aims to provide a new application of levofloxacin, and researches of the invention find that HP beta CD-Lev and Lev are administered to an ApoE-/-C57BL/6 mouse in a tail vein injection mode, the influence of the HP beta CD-Lev and Lev on high-fat diet modeling atherosclerosis is detected, and the drug effect of the HP beta CD-Lev on atherosclerosis resistance is determined, so that the levofloxacin can be used for delaying or treating vascular atherosclerosis diseases.

The invention is realized by the following technical scheme:

the application of levofloxacin in preparing antiatherosclerotic medicine has levofloxacin as the effective component.

Furthermore, the drug is encapsulated with levofloxacin through hydroxypropyl-beta-cyclodextrin.

Furthermore, the dosage of the levofloxacin is 10-80 mg/kg/2 d.

Compared with the prior art, the invention has the beneficial effects that:

(1) the invention defines the drug effect of Lev and HP beta CD-Lev in the aspect of anti-atherosclerosis through animal experiment level.

(2) The invention uses Lev and HP beta CD-Lev for delaying or treating cardiovascular diseases, particularly atherosclerosis diseases, expands the medication range of levofloxacin, reduces the cost of disease treatment, and has important research value and clinical application prospect.

(3) The invention can improve the water solubility of Lev and reduce the toxic and side effects of Lev by using HP beta CD to include Lev.

Drawings

FIG. 1 is a staining pattern of full-length vascular plaques of each experimental group in the examples;

FIG. 2 is a diagram illustrating quantitative detection of lipid droplets/plaques on the inner wall of the full-length aorta of each experimental group in the example, wherein the lesion area accounts for the percentage of the total aorta area;

FIG. 3 is an image of atherosclerotic plaque pathologically sectioned at the aortic valve site of each experimental group in the examples;

FIG. 4 is a graph showing the percentage of lesion area in the aortic root area in quantitative measurement of atherosclerotic plaque in the aortic valve region by each experimental group in the example.

Detailed Description

The invention will be further described with reference to the accompanying drawings.

Preparation examples: preparation of inclusion compound (HP beta CD-Lev) with HP beta CD including Lev

Putting a rotor of a magnetic stirrer into a 6ml glass bottle, accurately weighing 93mg Lev powder and 375mg HP beta CD powder by using a clean medicine spoon and an electronic analytical balance, pouring the weighed LEV and HP beta CD powder into the glass bottle, adding 5ml PBS into the glass bottle, covering a cover of the glass bottle, completely sealing the glass bottle in the dark by using tinfoil paper, fully dissolving and uniformly mixing the materials on the magnetic stirrer, and continuously stirring at a low speed for 48 hours to complete inclusion.

Removing a cover of the included HP beta CD-Lev, sealing a preservative film, freezing for 30min in a-80 refrigerator until the HP beta CD-Lev is completely solidified, simultaneously opening a low-temperature freeze dryer to pre-cool an instrument for 30min in advance, quickly taking out the HP beta CD-Lev after 30min, pricking holes on the sealed preservative film by using a needle of a 1ml syringe, then putting the HP beta CD-Lev into the freeze dryer, freeze-drying for 18h to completely remove water in the HP beta CD-Lev solution, fully drying, adding 5ml of trichloromethane, covering a glass bottle cap, fully dissolving the trichloromethane on a vortex oscillator, carrying out suction filtration on the HP beta CD-Lev solution in which the trichloromethane is dissolved by using a 0.22 mu m filter membrane, wherein Lev can be dissolved in the trichloromethane, but HP beta CD is insoluble in the trichloromethane, thus obtaining HP beta CD mucilaginous powder including Lev and trichloromethane solution in which the non-included Lev is dissolved respectively by suction filtration, collecting HP beta CD-Lev mucilage powder, fully drying in an oven for 5h, taking out the dried HP beta CD-Lev powder, filling the HP beta CD-Lev powder into an autoclaved 4ml EP tube, sealing the tube with a sealing film, and completely storing the tube in a dark place after being wrapped by tin foil paper.

Application examples

This example used high fat fed ApoE-/-male C57BL/6 mice as the atherosclerosis model mice.

The experiment was divided into 4 groups (6 mice per group) of: 1. control group (Normal ApoE not fed high fat)^-/-Mouse); 2. model group (high-fat fed ApoE injected with physiological saline)^-/-Mouse); 3. HP beta CD-Lev group (tail vein injection HP beta CD-Lev 61.94mg/kg/2 d); 4. lev group (tail vein injection Lev 61.94mg/kg/2d), wherein groups 2, 3, and 4 were continued for 12 weeks while being fed with high fat. The detection analysis was performed for each group of experimental mice as follows.

(1) Blood lipid detection

At 12 weeks post-dose, a blood sample was taken from the mice (mice should be fasted for 12 hours before sampling); centrifuging at 3000rpm for 10min to obtain serum sample; the Total Cholesterol (TC), Triglyceride (TG), high density lipoprotein-cholesterol (HDL-C) and low density lipoprotein-cholesterol (LDL-C) contents in the serum were measured using a fully automatic biochemical analyzer (Beckman Coulter AU 480).

The results show the following table 1, with normal ApoE without high fat feeding^-/-Mice comparison, high fat fed ApoE^-/-The levels of TG, TC, LDL-C and HDL-C of the mice are all obviously increased, which indicates that the blood fat of the mice is increased after high-fat feeding, and indicates that the initial success of molding is achieved. Significant reduction in TC and LDL-C levels following HP β CD-Lev, Lev administration (with high-fat fed ApoE)^-/-Mice showed significant differences). The result reflects that the HP beta CD-Lev and the HP beta CD-Lev both have the efficacy of inhibiting the rise of blood fat, and indirectly reflects the potential of resisting atherosclerosis.

TABLE 1 blood lipid levels in mice after HP β CD-Lev, Lev tail vein administration

The significant difference is as follows: a, P is less than 0.001, and the Model is compared with the Control group; comparing HP beta CD-Lev with Model group, p < 0.01;^##p < 0.01, Lev is compared to Model set.

(2) Quantitative detection of lipid droplets/plaques in the inner wall of full-length aorta

After 12 weeks of administration, the full length aorta (from the heel of aorta to the abdominal aorta to the iliac region) of the mice was fixed with 4% paraformaldehyde for 3 h; carefully removing the peripheral adipose tissues and longitudinally cutting open; dyeing for 0.5h with oil red O in isopropanol at room temperature; after 2-3 times of washing with 60% isopropyl alcohol, the dissected arterial blood vessel was laid on a glass slide with the inside facing up, covered with a cover glass to spread the blood vessel sufficiently, photographed, and the plaque was bright red, and then subjected to quantitative analysis.

The thoracic aorta was longitudinally dissected to expose the inside of the vessels, the plaques were stained with oil red O (red) as shown in FIG. 1, and the plaques were quantitatively analyzed as shown in FIG. 2, (. ANG., P < 0.01 comparing the Model group with the Control group;. # #., P < 0.01 comparing the HP. beta. CD-Lev group with the Model group;. #, P < 0.05 comparing the Lev group with the Model group).

As is apparent from FIG. 2, the thoracic aorta of normal ApoE-/-mice fed without high fat had almost no atherosclerotic plaques, whereas high fat-fed ApoE had little atherosclerotic plaques^-/-The mouse has a large number of red plaques (namely atherosclerotic plaques colored by oil red O) on the thoracic aorta, and the quantitative comparison shows that the atherosclerosis model of the mouse is successfully constructed. After the administration of HP beta CD-Lev and Lev, the area of the red plaque presents a significant difference compared with that of the Model group, and the LEV is proved to have the effect of resisting atherosclerosis.

(3) Imaging and quantification of atherosclerotic plaque at aortic valve sites

After 12 weeks of administration, the heart of the mouse connected with the aortic arch is taken, rapidly frozen and embedded with OCT tissue freezing agent; serial sections of 8 μm thickness were cut using a cryomicrotome (Leica CM 1520); fixing with 95% ethanol for 15 min; the staining with oil red O and hematoxylin was performed by conventional methods, and then photographed and analyzed.

In FIG. 3, (A-D) are image displays of pathological sections of each experimental group; FIG. 4 (E) shows the quantitative analysis of plaques (. lamda. sup. 0.001 compare Model to Control group. sup. 0.01, P. 0.01 compare Model to 0.01, compare HP, HP. 0.0.0.01 compare HP. g. compare HP. the HP. group to Model group, the HP. sup.

As is apparent from fig. 3 and 4, the intravascular side of the aortic valve of the normal ApoE-/-mice that were not given high fat had only very small atherosclerotic plaques, whereas the intravascular side of the aortic root of the ApoE-/-mice that were given high fat had very thick red plaques (i.e., oil red O-stained atherosclerotic plaques), which showed significant differences in quantitative comparisons, again indicating successful construction of an atherosclerosis model in mice. After the administration of the HP beta CD-Lev and Lev, the thickness and the area of the red plaque show significant difference compared with those of a Model group, and further prove that the HP beta CD-Lev and Lev have the effect of resisting atherosclerosis.

The foregoing merely represents preferred embodiments of the invention, which are described in some detail and detail, and therefore should not be construed as limiting the scope of the invention. It should be noted that, for those skilled in the art, various changes, modifications and substitutions can be made without departing from the spirit of the present invention, and these are all within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.