Composition, medicine and preparation method and application thereof

Abstract

The invention provides a composition, a medicine and a preparation method and application thereof, the composition relates to the technical field of medicine, the composition comprises a carrier and an active component loaded on the carrier, the carrier contains cell wall peptidoglycan, the active component comprises a first active component with a specific treatment effect on a focus, and the first active component comprises a second active component with a specific treatment effect on the focus. And a second active ingredient having affinity to the lesion area, wherein the first active ingredient comprises bile acid; the second active ingredient includes a targeting peptide. The composition disclosed by the invention can activate endothelial cells in a targeted manner, and can reduce various heart and brain diseases, metabolic diseases, infectious diseases, allergic diseases and the like caused by endothelial cell dysfunction, so that diseases and death caused by the dysfunction are reduced.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, and in particular to a composition. Furthermore, this invention also relates to a drug prepared using the composition, a method for preparing the composition, and the use of the composition. Background Technology

[0002] Endothelial cell dysfunction refers to the disorder of the structure and function of vascular endothelial cells under the influence of various internal and external factors, resulting in abnormalities in vasomotor function, permeability, and coagulation and anticoagulation balance. This abnormality is an important pathological basis for many cardiovascular and cerebrovascular diseases, metabolic diseases, allergic diseases, and infectious diseases, and is also an important cause of disease and death.

[0003] Endothelial cells are a thin layer of flattened epithelial cells, tightly packed together, forming part of the blood vessel wall. Endothelial cells are widely distributed on the inner walls of all blood vessels, including the heart, arteries, capillaries, and veins, forming a continuous single-cell layer.

[0004] Although endothelial cells exist as a single-celled layer in all organs, the vascular endothelium is currently considered a dynamic and interconnected whole. Endothelial cell function has a decisive impact on organ function, systemic health, and healthy aging.

[0005] Inflammation is a key determinant of many human diseases, and endothelial cells are highly involved in and mediate the development of inflammation-related diseases, such as cardiovascular diseases, neurological diseases, metabolic diseases, infectious diseases, allergic diseases, and other diseases affecting multiple tissues and organs.

[0006] Under pathological conditions, endothelial cell inflammation leads to increased production of inflammatory factors, and key inflammatory enzymes such as cyclooxygenase-2 (COX-2) are induced to overexpress, resulting in increased production of prostaglandins such as PGE2, further causing damage. Currently, there are no effective interventions for endothelial cell inflammation and related diseases. Summary of the Invention

[0007] In view of this, the present invention aims to provide a composition suitable for preparing a medicine for treating endothelial cell inflammatory damage and related diseases.

[0008] To achieve the above objectives, the technical solution of the present invention is implemented as follows:

[0009] A composition, characterized in that:

[0010] The composition includes a carrier and a first active ingredient and a second active ingredient loaded on the carrier;

[0011] The carrier contains cell wall peptidoglycan, the first active ingredient includes bile acids, and the second active ingredient includes targeting peptides.

[0012] Furthermore, the bile acids include at least one of tauroursodeoxycholic acid, ursodeoxycholic acid, chenodeoxycholic acid, and porcine deoxycholic acid.

[0013] Furthermore, the targeting peptide includes a targeting peptide for targeting inflammation; or, the targeting peptide includes a targeting peptide for targeting endothelial cells.

[0014] Furthermore, the vector includes Bacillus.

[0015] Furthermore, the carrier is covalently connected to both the first active ingredient and the second active ingredient.

[0016] Compared with the prior art, the present invention has the following advantages:

[0017] The composition described in this invention can target inflammatory activation of endothelial cells, thereby reducing various cardiovascular diseases, neurological diseases, metabolic diseases, allergic and infectious diseases caused by endothelial cell dysfunction, and thus reducing the diseases and deaths caused by such abnormalities.

[0018] Another object of the present invention is to provide a medicine comprising the composition described above in its preparation components.

[0019] The pharmaceutical product of the present invention, by applying the above-described composition, achieves the same beneficial effects as the prior art, which will not be elaborated further here.

[0020] Meanwhile, another object of the present invention is to provide a method for preparing the above composition, the method comprising:

[0021] Take activated cell wall peptidoglycan, bile acids and target peptides and add them to buffer solution to react fully;

[0022] The product obtained from the reaction was centrifuged with deionized water and then freeze-dried to obtain the composition.

[0023] Furthermore, another object of the present invention is to provide the use of the above composition in the preparation of pharmaceuticals.

[0024] Furthermore, the drug includes a drug for treating vascular endothelial cell damage, or a drug for treating metabolic diseases, or a drug for treating allergic diseases, or a drug for treating infectious diseases, or a drug for treating neurological diseases.

[0025] Furthermore, the medicine includes medicines for treating heart injury, or medicines for treating brain injury. Attached Figure Description

[0026] The accompanying drawings, which form part of this invention, are used to provide a further understanding of the invention. The illustrative embodiments of the invention and their descriptions are used to explain the invention and do not constitute an undue limitation of the invention. In the drawings:

[0027] Figure 1 Image showing the inhibitory effect of double-labeled immunofluorescence on COX-2, a key enzyme in vascular endothelial inflammation;

[0028] Figure 2 Images showing the improvement in myocardial morphology under a microscope;

[0029] Figure 3 Images showing the detection of cerebral vascular permeability using FITC-glucan;

[0030] Figure 4 This is a schematic diagram of the relative alternation rate tested using Y-maze behavioral analysis.

[0031] Figure 5 This is a schematic diagram of the relative sniffing time tested using a three-box behavioral experiment. Detailed Implementation

[0032] It should be noted that, unless otherwise specified, the embodiments and features described in the present invention can be combined with each other.

[0033] The present invention will now be described in detail with reference to the accompanying drawings and embodiments.

[0034] This embodiment relates to a composition that can effectively treat diseases caused by abnormal vascular endothelial cell function.

[0035] Specifically, the composition of this embodiment includes a carrier and an active ingredient loaded on the carrier, wherein the carrier is a carrier containing cell wall peptidoglycan, and the active ingredient includes a first active ingredient that has a specific therapeutic effect on the lesion and a second active ingredient that has an affinity for the lesion area.

[0036] More specifically, the first active ingredient includes a drug containing -NH2, -OH, or -COOH, while the second active ingredient includes a targeting substance containing -NH2, -OH, or -COOH.

[0037] In a preferred embodiment, the drug includes bile acids, preferably bile acids that have a specific therapeutic effect on the lesion, that is, bile acids with beneficial effects. For example, bile acids include at least one of the following bile acids with beneficial effects: tauroursodeoxycholic acid (TUDCA), ursodeoxycholic acid (UDCA), chenodeoxycholic acid (CDCA), and hyodeoxycholic acid (HDCA).

[0038] It should be understood that since TUDCA, UDCA, CDCA and HDCA all contain -NH2 or -OH or -COOH, these bile acids play the same role in the preparation of the composition and in the specific treatment of the lesion area. Therefore, the following explanation will use TUDCA as an example of bile acid.

[0039] It should be noted that bile acids are synthesized from cholesterol in the liver into primary bile acids (cholic acid and chenodeoxycholic acid), which are then secreted into the intestines. There, they are acted upon by bacteria to produce secondary bile acids (deoxycholic acid and lithocholic acid), promoting the digestion and absorption of lipids. 95% of intestinal bile acids are reabsorbed into the bloodstream, with the majority entering the liver and a small portion entering the blood circulation. In the liver, bile acids combine with taurine or glycine to form hydrophilic conjugated bile acids, which then re-enter the enterohepatic circulation.

[0040] Recent studies have shown that bile acids, as signaling molecules, can exert multifaceted regulatory effects on various tissues and organs through blood circulation. Tauroursodeoxycholic acid (TUDCA), the main component of the traditional drug bear bile, is a conjugated bile acid formed by the combination of taurine and ursodeoxycholic acid. Compared to ursodeoxycholic acid, the hydrophilic TUDCA has more routes of administration, lower toxicity to tissues and organs, and wider applications. Dr. Hao Li and colleagues from the Swiss Federal Institute of Technology in Lausanne (EPFL) also emphasized the importance of the protective effect of TUDCA in an article published in *Cell Metab*.

[0041] Current research has found that TUDCA can alleviate endoplasmic reticulum stress, reduce inflammation, fight oxidation, and inhibit apoptosis, effectively antagonize endothelial cell damage, and play a potential therapeutic role in various diseases such as cardiovascular disease, diabetes, and neuropathy.

[0042] However, since the target metabolic organs for bile acids are the liver and intestines, TUDCA accumulates in hepatocytes and undergoes enterohepatic circulation regardless of the administration route. This results in either excessively low blood concentrations of TUDCA, failing to effectively counteract damage to other tissues and organs, or excessively high intrahepatic concentrations, causing damage to hepatocytes. This severely limits the widespread clinical application of TUDCA in extrahepatic tissues and organs. However, the development of targeted drug delivery methods has brought new hope for the broader clinical application of TUDCA.

[0043] In a preferred embodiment, the targeting substance includes a targeting peptide, which may include only targeting peptides that target inflammation. Specifically, the targeting peptides that target inflammation are those that target endothelial cells activated by inflammation. Alternatively, the targeting peptide may include only targeting peptides that target endothelial cells. In the following description, VHPK peptides will be used as an example of a targeting peptide; it should be understood that targeting peptides include, but are not limited to, VHPK peptides.

[0044] Since cell wall peptidoglycan is an important component of bacterial cell walls and is widely found in spore-forming probiotics, in a preferred embodiment, cell wall peptidoglycan includes spore cell wall peptidoglycan. The spore cell wall is the outer structure of the dormant spore, a special type of bacterial organism, and possesses extremely high resistance and stability. Spore cell wall peptidoglycan is simply the peptidoglycan found in the spore cell wall.

[0045] Therefore, as a preferred embodiment, the carrier of this embodiment includes spore-forming probiotics, and the spore-forming probiotics include at least one of Bacillus subtilis (accession number 1511C0002100009606, China General Microbiological Culture Collection Center), BA (Bacillus amyloliquefaciens spore; accession number 1511C0002100009607, China General Microbiological Culture Collection Center), and BM (accession number 1511C0002100009511, China General Microbiological Culture Collection Center).

[0046] In this embodiment, natural probiotics are used as a carrier, which is harmless to the human body, making it possible to widely apply bioactive bile acids and other substances in extrahepatic tissues and organs. Moreover, the carrier is safe and effective, making the composition of this embodiment applicable to the treatment of diseases caused by endothelial cell dysfunction.

[0047] For ease of understanding, it's important to note that Bacillus is a bacterium with probiotic activity and health-promoting effects. Our previous research found that Bacillus amyloliquefaciensspore (BA) spore peptidoglycan, as a nanoparticle-based drug loading material, exhibits high safety and good biocompatibility.

[0048] BA carriers are primarily degraded within blood vessels via phagocytosis by monocytes and other cells, thus exhibiting no cytotoxicity and providing long-circulating drug release. BA nanocarriers have small, uniform particle sizes (<700 nm) and can be controlled in size, allowing for the loading of various drugs and targeting substances to achieve precise treatment. Similarly, peptidoglycans from the well-tolerated and safe Bacillus megaterium (BM) can also serve as carriers for bile acid drugs, targeting inflammatory damage to vascular endothelial cells.

[0049] It should be noted that since Bacillus plays the same role in preparing the composition of this embodiment as it does in treating diseases caused by endothelial cell dysfunction, the following description will use Bacillus BM as an example. However, it should be understood that probiotics other than Bacillus BM can also be used, such as Bacillus BA, Bacillus subtilis, and other spore-forming probiotics.

[0050] Since va-histone-proline-lysine (VHPK) peptide can target and bind to inflammatory-activated endothelial cells, in this embodiment, a TUDCA-BM-VHPK drug complex targeting inflammatory-activated endothelial cells was prepared by loading TUDCA and VHPK onto BM, in order to target and intervene in endothelial cell inflammatory damage and related diseases.

[0051] In a preferred embodiment, the aforementioned carrier is covalently linked to both the first and second active ingredients, and the covalent bonds are formed via the carbodiimide method. The size of the complex is between 1 nanometer and 1000 nanometers, and the aforementioned carrier is preferably a cell wall peptidoglycan with a size between 1 nanometer and 800 nanometers.

[0052] The composition of this invention is a bile acid drug that can target inflammatory activated endothelial cells. It contains probiotic BM peptidoglycan for targeting the drug carrier, pharmaceutically acceptable bile acid active drug components with anti-inflammatory effects such as TUDCA, and VHPK peptides that target inflammatory activated endothelial cells to improve inflammatory endothelial damage and related diseases. It can solve the problem in the prior art that the main target organs of bile acids are the liver and gallbladder, which limits their effective application in extrahepatic tissues and organs. It provides a new treatment for endothelial cell inflammatory damage and related cardiovascular, neurological, metabolic, allergic and infectious diseases.

[0053] Preferably, inflammatory endothelial injury includes, but is not limited to, the aorta, and large, medium, small, and microvessels of various tissues and organs.

[0054] It should be noted that the composition of this embodiment can be used for targeting and intervention of lipopolysaccharide-stimulated inflammatory endothelial cells in the aorta.

[0055] Preferably, inflammatory endothelial injury includes, but is not limited to, inflammatory vascular endothelial injury caused by stimulation from lipopolysaccharides, hyperglycemia, hyperlipidemia, hyperuricemia, pathogen infection, and allergens.

[0056] Preferably, the indicators of inflammatory endothelial injury include, but are not limited to, inflammatory factors, COX-2, PGE2, and reactive oxygen species.

[0057] Another object of the present invention is to provide a medicine comprising the above-described composition in its preparation components.

[0058] The pharmaceutical product of the present invention, by applying the above-described composition, achieves the same beneficial effects as the prior art, which will not be elaborated further here.

[0059] Specifically, the preparation method of the composition in this embodiment includes:

[0060] S1. Take the activated cell wall peptidoglycan, bile acids and target peptides and add them to the buffer solution to react fully;

[0061] S2. The product obtained from the reaction is centrifuged with deionized water and then freeze-dried to obtain the composite.

[0062] It should be noted that, in the preferred embodiment, the raw materials for preparing the composition of this embodiment contain 10 6 In the pure activated spore cell wall peptidoglycan prepared by a Bacillus vector, the amount of targeted peptide added is between 0.04 mg and 0.06 mg, such as 0.04 mg, 0.05 mg, 0.06 mg, etc., and the amount of bile acid added is between 0.9 mg and 1.1 mg, such as 0.9 mg, 1.0 mg, 1.1 mg, etc.

[0063] In a preferred embodiment, the weight ratio of the added targeting peptide to bile acid is between 4:100 and 6:100, such as 4:100, 5:100, 6:100, etc.

[0064] The BM-TUDCA drug complex prepared by the above steps can be used to detect whether TUDCA has been successfully linked to the BM carrier by Fourier transform infrared spectroscopy and thermogravimetric analysis.

[0065] In one exemplary embodiment, the preparation method of the aforementioned composition includes collecting and inactivating spores, reacting them with a purified solution (0.1 mol / L NaCl, 0.1 mol / L NaOH, 1% SDS (Sodium Dodecyl Sulfate), 0.1 mol / L DTT (Dithiothreitol)) under magnetic stirring at 70°C for 2 h, and washing the spores five times with double-distilled water to obtain a purified spore peptidoglycan carrier.

[0066] The purified 10 was obtained by the carbodiimide method. 6 One Bacillus megaterium (BM) spore peptidoglycan carrier was reconstituted in 1 mL of MES (2-(N-morpholino)ethanesulfonic acid, 0.1 mol / mL, pH 5.5) buffer. Excess EDC and NHS were added sequentially to activate the free carboxyl groups, yielding pure activated bacterial spore cell wall peptidoglycan. Then, 1 mg of the anti-inflammatory bile acid TUDCA and 0.05 mg of VHPK peptide were added to link TUDCA, VHPK, and BM, resulting in a pure TUDCA drug complex targeting inflammatory activated endothelial cells, which is the composition of this embodiment.

[0067] Fourier transform infrared spectroscopy and thermogravimetric analysis were used to determine the conjugation efficiency and drug loading. The drug loading of TUDCA was found to be 10%-30%, and the conjugation rate of the inflammation-activating endothelial cell-targeting peptide VHPK was approximately 90%. The following experiments will use this complex as an example to verify its effect on diseases caused by endothelial cell dysfunction.

[0068] In one exemplary embodiment, the method for obtaining activated cell wall peptidoglycan specifically includes:

[0069] S001. Take 200 μL of Bacillus megaterium (BM) culture stored at -20℃ with 50% glycerol, add it to 5 mL of sterilized LB liquid medium, and incubate overnight at 180 rpm in a temperature-controlled shaker at 37℃ to complete the activation of the culture.

[0070] S002. Take 200 μL of the activated bacterial solution and add it to the cooled LB solid medium plate. Spread it evenly with a glass rod and invert the plate in a 37°C constant temperature incubator for 7 days. BM begins to grow and transform into spores. Collect the BM spores that have grown on the plate, wash the spore suspension 3 times with double-distilled water, and then sonicate it 5 times under ice bath conditions, 3 minutes each time, with an interval of 6 minutes between each sonication to prevent the suspension from being overheated and affecting the spores.

[0071] S003. After completing the sonication treatment of the suspension, centrifuge and wash three times to collect the precipitated spores. Resuspend the spores in a certain volume of 1% trypsin solution and shake overnight at 80 rpm in a shaker at 37°C. Centrifuge to remove the trypsin solution and add a certain amount of purification solution (see the description above for the specific purification solution). Stir magnetically at 70°C for 2 hours. Centrifuge the treated spores to remove the supernatant and wash the spores five times with double-distilled water. Resuspend the spores again and take a small amount for spore counting. The remaining spores are inactivated by autoclaving at 121°C for 30 minutes to remove their activity, thus obtaining the purified spore peptidoglycan carrier.

[0072] S004, The 6×10⁶ particles obtained are processed using the carbodiimide method. 6 One BM spore carrier was reconstituted in 1 mL of 0.1 mol / mL pH 5.5 MES (2-(N-morpholino)ethanesulfonic acid) buffer. Excess EDC (1-ethyl-(3-dimethylaminopropyl)carbodiimide) and NHS (N-hydroxysuccinimide) were added sequentially until saturation. The molar ratio of EDC to NHS was 1.5:1, and the catalyst should be added in excess. The mixture was placed on a shaker and activated at 4°C for 60 minutes to activate the free carboxyl groups. The spores were washed with double-distilled water and centrifuged at 12,000 rpm for 3 minutes. This process was repeated 5 times to obtain activated spores.

[0073] It should be noted that, in the preferred embodiment, in step S1 above, the activated cell wall peptidoglycan and bile acids are added to the buffer solution simultaneously, followed by the targeting peptide, with an interval of 15 min to 30 min. In this step, the buffer solution is a PBS solution with a pH of 7.2.

[0074] In step S2 above, deionized water is used for centrifugation. Specifically, the centrifugation speed is preferably between 10,000 rpm and 15,000 rpm, such as 10,000 rpm, 11,000 rpm, 12,000 rpm, 13,000 rpm, 14,000 rpm, 15,000 rpm, etc. The centrifugation time is 3 min to 5 min, such as 3 min, 4 min, or 5 min, and it is repeated 3 to 5 times, such as 3 times, 4 times, or 5 times.

[0075] As an exemplary embodiment, the preparation method of the composition of this embodiment includes the following preparation steps: reconstitute the activated spores in 1 mL of PBS buffer (0.1 mol / L, pH 7.2), add 1 mg of anti-inflammatory bile acid TUDCA, place the mixture on a shaker, and react at room temperature for 20 minutes. Then add 0.05 mg of VHPK peptide, and activate at room temperature for 100 minutes to allow the primary amino group to react with the activated carboxyl group to form an amide bond. Connect TUDCA, VHPK, and BM. Wash with double-distilled water and centrifuge three times to remove unreacted UK and RGD. Collect the microspheres and freeze-dry them to obtain a pure TUDCA drug complex targeting inflammatory activated endothelial cells, which is the composition of this embodiment.

[0076] Next, we examined the intervention effect of the VHPK-BM-TUDCA targeted drug complex in this embodiment on lipopolysaccharide-stimulated inflammatory endothelial injury in mice.

[0077] Figure 1 After double-labeling with immunofluorescence, the inhibitory effect of VHPK-BM-TUDCA on COX-2, a key enzyme in lipopolysaccharide-stimulated vascular endothelial inflammation in mice, was shown under a fluorescence microscope.

[0078] Thirty-two Kunming mice were selected and divided into a control group, a lipopolysaccharide group, a lipopolysaccharide + TUDCA group, and a lipopolysaccharide + VHPK-BM-TUDCA group.

[0079] In the lipopolysaccharide (LPS) group, an inflammatory stimulation model was established by intraperitoneal injection of LPS 1 mg / kg / d five times. The LPS + TUDCA group received intraperitoneal injection of TUDCA 300 mg / kg / d, and the LPS + VHPK-BM-TUDCA group received VHPK-BM-TUDCA 10 mg / kg / d via tail vein injection. 4 The mice were injected once every two days. After three injections, the mice were sacrificed for tissue collection. Vascular tissue was collected, frozen sections were prepared, and double-labeled immunofluorescence was used to visualize the content and distribution of COX-2 in the vascular endothelium. Red fluorescence was used to visualize the content and distribution of COX-2, and green fluorescence was used to visualize the distribution of the vascular endothelial marker molecule CD31.

[0080] The results showed that, compared with the normal control group, the lipopolysaccharide (LPS) group had a significantly increased COX-2 content in the vascular endothelium. TUDCA intervention could partially inhibit the distribution of COX-2 content, and VHPK-BM-TUDCA could almost completely inhibit the distribution of COX-2 content. These results indicate that VHPK-BM-TUDCA can effectively inhibit the distribution of COX-2, a key enzyme in vascular endothelial inflammation stimulated by LPS.

[0081] Figure 2 Microscopic observation showed that VHPK-BM-TUDCA improved the morphology of lipopolysaccharide-stimulated mouse myocardium.

[0082] Thirty-two Kunming mice were selected and divided into a control group, a lipopolysaccharide group, a lipopolysaccharide + TUDCA group, and a lipopolysaccharide + VHPK-BM-TUDCA group.

[0083] In the lipopolysaccharide (LPS) group, an inflammatory stimulation model was established by intraperitoneal injection of LPS 1 mg / kg / d five times. The LPS + TUDCA group received intraperitoneal injection of TUDCA 300 mg / kg / d, and the LPS + VHPK-BM-TUDCA group received VHPK-BM-TUDCA 10 mg / kg / d via tail vein injection. 4 The mice were injected once every two days. After three injections, the mice were sacrificed for tissue collection. Heart tissue was collected, sectioned, and the morphological changes of the myocardium were observed using HE staining (hematoxylin-eosin staining) and microscopy.

[0084] The results showed that, compared with the normal control group, the lipopolysaccharide group exhibited endothelial shedding and disordered myocardial fiber dissolution. TUDCA intervention partially improved the morphological changes in the myocardium, and VHPK-BM-TUDCA significantly improved the morphological changes in the myocardium, almost approaching the morphology of the normal group. These results indicate that VHPK-BM-TUDCA can significantly improve the morphological changes in the myocardium stimulated by lipopolysaccharide.

[0085] Next, we examined the effect of the VHPK-BM-TUDCA targeted drug complex in this embodiment on improving the blood-brain barrier and brain function.

[0086] Figure 3 The FITC-glucan method was used to detect the effect of VHPK-BM-TUDCA on improving cerebral vascular permeability in mice stimulated by lipopolysaccharide.

[0087] Thirty-two Kunming mice were selected and divided into a control group, a lipopolysaccharide group, a lipopolysaccharide + TUDCA group, and a lipopolysaccharide + VHPK-BM-TUDCA group.

[0088] In the lipopolysaccharide (LPS) group, an inflammatory stimulation model was established by intraperitoneal injection of LPS 1 mg / kg / d five times. The LPS + TUDCA group received intraperitoneal injection of TUDCA 300 mg / kg / d, and the LPS + VHPK-BM-TUDCA group received VHPK-BM-TUDCA 10 mg / kg / d via tail vein injection. 4 The injections were administered every two days. After three injections, the Y-maze and three-box behavioral tests were used to assess changes in cognitive, social, and other brain functions. Following the behavioral tests, the permeability of the blood-brain barrier was measured using fluorescein isothiocyanate (FITC)-glucan 70kD.

[0089] Detection of blood-brain barrier permeability using the fluorescent probe FITC-glucan method: Mice were injected intravenously via the tail vein with 10 mg of FITC-glucan dissolved in 0.1 mL of physiological saline. Thirty minutes later, the animals were sacrificed, and the brains were quickly removed and frozen. Frozen sections were observed and photographed under a fluorescence microscope.

[0090] The results showed that almost no FITC fluorescence was detected in the control group, while the fluorescence intensity of the vascular leakage was significantly enhanced in the lipopolysaccharide model group. After TUDCA intervention, the leakage fluorescence was significantly reduced, with the reduction being most significant in the lipopolysaccharide + targeted TUDCA group.

[0091] Figure 4 and Figure 5 The study demonstrated the ameliorative effect of VHPK-BM-TUDCA on lipopolysaccharide-stimulated behavioral abnormalities in mice, as assessed using the Y-maze and three-box behavioral tests.

[0092] Thirty-two Kunming mice were selected and divided into a control group, a lipopolysaccharide group, a lipopolysaccharide + TUDCA group, and a lipopolysaccharide + VHPK-BM-TUDCA group.

[0093] In the lipopolysaccharide (LPS) group, an inflammatory stimulation model was established by intraperitoneal injection of LPS 1 mg / kg / d five times. The LPS + TUDCA group received intraperitoneal injection of TUDCA 300 mg / kg / d, and the LPS + VHPK-BM-TUDCA group received VHPK-BM-TUDCA 10 mg / kg / d via tail vein injection. 4 The injections were administered every two days. After three injections, the Y-maze and three-box behavioral tests were used to assess changes in cognitive, social, and other brain functions. Following the behavioral tests, the permeability of the blood-brain barrier was measured using FITC-glucan 70kD.

[0094] The results showed that, compared with the normal control group, the cognitive and social functions of mice in the lipopolysaccharide group were significantly inhibited, while TUDCA intervention could improve behavioral abnormalities. Moreover, VHPK-BM-TUDCA significantly improved behavioral abnormalities, approaching the level of the normal group. The results indicate that VHPK-BM-TUDCA can significantly improve the behavioral changes in mice stimulated by lipopolysaccharide.

[0095] Furthermore, another object of the present invention is to provide the use of the above composition in the preparation of pharmaceuticals.

[0096] In one preferred embodiment, the medicine includes a medicine for treating vascular endothelial cell injury, and more preferably, the medicine includes a medicine for treating lipopolysaccharide-induced inflammatory endothelial cell injury.

[0097] In one preferred embodiment, the medicine includes a drug for treating metabolic, allergic, infectious, and neurological diseases. Preferably, the medicine includes a drug for treating metabolic, allergic, infectious, and neurological diseases associated with inflammatory endothelial cell damage.

[0098] In one preferred embodiment, the pharmaceutical product includes a medicine for treating cardiac injury, preferably, the medicine includes a medicine for treating cardiac disease associated with inflammatory endothelial cell injury. Preferably, cardiac injury associated with inflammatory endothelial cell injury includes, but is not limited to, pathological changes and functional changes in blood vessels and myocardium.

[0099] In one preferred embodiment, the medicament includes a pharmaceutical product for treating brain injury. Preferably, the medicament includes a treatment for brain diseases associated with inflammatory endothelial cell injury. Preferably, brain injury associated with inflammatory endothelial cell injury includes, but is not limited to, changes in cerebral vascular permeability, brain morphology, and behavior.

[0100] It should be noted that the aforementioned drugs include those used to treat cardiovascular and cerebrovascular injuries. Preferably, the drugs include those used to treat cardiovascular and cerebrovascular diseases associated with inflammatory endothelial cell injury. It should be noted that cardiovascular and cerebrovascular diseases associated with inflammatory endothelial cell injury include, but are not limited to, metabolic, infectious, and allergic cardiovascular and neurological diseases. In addition, the aforementioned drugs may also be used to treat inflammatory-related diseases of other systems throughout the body.

[0101] The above are only preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A composition, characterized in that: The composition comprises a carrier and a first active ingredient and a second active ingredient loaded on the carrier; The carrier contains cell wall peptidoglycan, the first active ingredient includes bile acid, and the second active ingredient includes a targeting peptide.

2. The composition according to claim 1, characterized in that: The bile acid includes at least one of tauroursodeoxycholic acid, ursodeoxycholic acid, chenodeoxycholic acid and hyodeoxycholic acid.

3. The composition according to claim 1, characterized in that: The targeting peptide includes a targeting peptide for targeting inflammation; or, the targeting peptide includes a targeting peptide for targeting endothelial cells.

4. The composition according to claim 1, characterized in that: The vector comprises Bacillus.

5. The composition according to any one of claims 1 to 4, characterized in that: The carrier is connected to both the first active ingredient and the second active ingredient via covalent bonds.

6. A medicine, characterized in that: The preparation components of the medicine include the composition according to any one of claims 1-5.

7. A method for preparing the composition according to any one of claims 1 to 5, characterized in that: The method comprises: Take activated cell wall peptidoglycan, bile acid and targeting peptide and add them into buffer for full reaction; The product obtained by the reaction is centrifuged with deionized water, and then freeze-dried to obtain the composition.

8. Use of the composition according to any one of claims 1 to 5 in the preparation of medicines.

9. The use according to claim 8, characterized in that: The medicine includes medicine for treating vascular endothelial cell damage, or, the medicine includes medicine for treating metabolic diseases, or, the medicine includes medicine for treating allergic diseases, or, the medicine includes medicine for treating infectious diseases, or, the medicine includes medicine for treating neurological diseases.

10. The use according to claim 8, characterized in that: The medicine includes a medicine for treating heart damage, or, the medicine includes a medicine for treating brain damage.

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