A pharmaceutical composition comprising β-hydroxybutyrate and vitamin D and its uses

Abstract

The present invention provides the use of a pharmaceutical composition in the preparation of a medicament for treating diseases associated with high permeability of the blood-brain barrier and intestinal barrier. The pharmaceutical composition contains pharmaceutically acceptable β-hydroxybutyrate and vitamin D as active ingredients. In classical systemic inflammations of nerves, intestines, etc. induced by bacterial endotoxin lipopolysaccharide (LPS), the pharmaceutical composition of the present invention can well reduce the high permeability of the blood-brain barrier and intestinal barrier, and play a role in treating central nervous system inflammation, nerve damage, behavioral abnormalities, etc.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical compositions, and more particularly, to a pharmaceutically acceptable pharmaceutical composition of β-hydroxybutyrate and vitamin D and its use. Background Art

[0002] The blood-brain barrier is a semi-permeable barrier between the blood circulation system and the central nervous system, which is of great significance for maintaining the relative stability of the physical and chemical factors in the brain tissue. The high permeability of the blood-brain barrier causes cerebral edema, inflammation, etc., and a series of neurological deficit symptoms, which is a common factor in the occurrence of various neurological diseases. At the same time, the intestinal barrier is also a selectively permeable barrier, which supports nutrient absorption and waste secretion while preventing the invasion of luminal substances. The high permeability of the intestinal barrier allows pathogenic factors such as bacteria and toxins to enter the blood circulation and penetrate into the central nervous system through the damaged blood-brain barrier, causing pathological damage such as inflammation, which is a common factor in the occurrence of various neurological diseases such as autism, depression, anxiety, Alzheimer's disease, Parkinson's disease, etc. In addition, the high permeability of the intestinal barrier is also related to certain pathologies and aging, and is also the cause of bacterial infections, systemic and liver inflammation, food allergies and autoimmune diseases.

[0003] However, at present, due to the high permeability of the blood-brain barrier and the intestinal barrier, various diseases such as neuroinflammation, nerve damage, and related inflammations, allergies, and autoimmune diseases in other systems of the body generally have no good intervention clinically and the prognosis is not good. Therefore, there is an urgent need to develop drugs with better curative effects to improve the high permeability of the blood-brain barrier and the intestinal barrier, so as to treat diseases related to the high permeability of the blood-brain barrier and the intestinal barrier, especially neurological diseases. Summary of the Invention

[0004] Technical Problem

[0005] An object of the present invention is to provide a pharmaceutical composition comprising a pharmaceutically acceptable β-hydroxybutyrate and vitamin D as active ingredients for improving the high permeability of the blood-brain barrier and the intestinal barrier, thereby treating diseases related to the high permeability of the blood-brain barrier and the intestinal barrier, so as to solve the problems in the prior art that due to the high permeability of the blood-brain barrier and the intestinal barrier, various diseases such as neuroinflammation, nerve damage, and related inflammations, allergies, and autoimmune diseases in other systems of the body generally have no good intervention clinically and the prognosis is not good.

[0006] Another object of the present invention is to provide a pharmaceutical composition comprising a pharmaceutically acceptable β-hydroxybutyrate and vitamin D as active ingredients. β-hydroxybutyrate and vitamin D are food-grade drugs with high safety, and the combined use of drugs has a sensitizing effect and the effect is more obvious.

[0007] Technical solution

[0008] According to one aspect of the present invention, there is provided a pharmaceutical composition comprising a pharmaceutically acceptable β-hydroxybutyrate and vitamin D as active ingredients.

[0009] In one example, in the pharmaceutical composition of the present invention, the content of β-hydroxybutyrate is 8,000 - 30,000 parts by weight, and the content of vitamin D is 0.8 - 1 part by weight; preferably, the amount of β-hydroxybutyrate is 8,000 - 20,000 parts by weight, and the amount of vitamin D is 0.08 - 0.5 part by weight; more preferably, the amount of β-hydroxybutyrate is 10,000 - 15,000 parts by weight, and the amount of vitamin D is 0.1 - 0.5 part by weight; most preferably, the amount of β-hydroxybutyrate is 10,000 parts by weight, and the amount of vitamin D is 0.1 part by weight. Preferably, the above contents of β-hydroxybutyrate and vitamin D are based on the content for human use, and those skilled in the art can also convert them to the content for mouse use.

[0010] Preferably, the β-hydroxybutyrate includes sodium salt, calcium salt and magnesium salt of pharmaceutically acceptable β-hydroxybutyric acid.

[0011] Preferably, the vitamin D includes: vitamin D3, vitamin D2, alfacalcidol, calcifediol (25-hydroxyvitamin D), calcitriol (1α,25-dihydroxyvitamin D), dihydrotachysterol (DHT) or a combination thereof; preferably, the vitamin D is active vitamin D3.

[0012] In another example, the pharmaceutical composition according to the present invention further comprises a pharmaceutically acceptable solvent; preferably, the pharmaceutically acceptable solvent is selected from one or more of sesame oil, olive oil, soybean oil, corn oil, grape seed oil, evening primrose oil and sunflower oil.

[0013] In yet another example, the pharmaceutical composition according to the present invention further comprises pharmaceutically acceptable excipients (such as sugar alcohols such as lactose and starch as fillers), lubricants (such as magnesium stearate and calcium stearate), plasticizers, disintegrants and preservatives, etc., which can be specifically selected according to the properties of the pharmaceutical composition and the dosage form to be prepared, etc.

[0014] Preferably, the pharmaceutical composition according to the present invention is made into a pharmaceutically acceptable dosage form; preferably, the dosage form includes tablets, capsules, granules, disintegrants, dispersants, pills, oral liquids and films.

[0015] According to another aspect of the present invention, the present invention provides the use of the above-mentioned pharmaceutical composition in the preparation of a drug for treating diseases related to the high permeability of the blood-brain barrier and intestinal barrier.

[0016] Preferably, the diseases associated with the high permeability of the blood-brain barrier and intestinal barrier include: neurological diseases and inflammatory and allergic diseases of other systems of the body; preferably, the neurological diseases include autism, depression, anxiety, Alzheimer's disease, and Parkinson's syndrome.

[0017] Preferably, the above-mentioned use is to reduce the high permeability of the blood-brain barrier and intestinal barrier caused by systemic inflammation, thereby alleviating nerve and systemic damage.

[0018] Beneficial effects

[0019] The pharmaceutical composition provided by the present invention uses β-hydroxybutyrate and vitamin D as the pharmaceutically active ingredients, and β-hydroxybutyrate and vitamin D have a synergistic effect. In the classical systemic inflammation of nerves, intestines, etc. induced by bacterial toxin lipopolysaccharide (LPS), the pharmaceutical composition of the present invention can well reduce the high permeability of the blood-brain barrier and intestinal barrier, and play a role in treating central nervous system inflammation, nerve damage, and behavioral abnormalities.

[0020] The pharmaceutical composition of the present invention uses food-grade drugs, β-hydroxybutyrate and vitamin D as the pharmaceutically active ingredients. In terms of alleviating the high permeability of the blood-brain barrier and intestinal barrier, it has high safety, strong sensitization effect in combination, and obvious therapeutic effect. Description of the drawings

[0021] Figure 1 Shows the results of the open field test behavioral detection on the 14th day of drug administration using the pharmaceutical composition according to an embodiment of the present invention (1: control group; 2: sodium β-hydroxybutyrate group; 3: vitamin D3 group; 4: lipopolysaccharide model group; 5: lipopolysaccharide model + sodium β-hydroxybutyrate group; 6: lipopolysaccharide model + sodium β-hydroxybutyrate + vitamin D3 group; 7: lipopolysaccharide model + vitamin D3 group); **P < 0.01, relative to the control group; #P < 0.05, relative to the lipopolysaccharide model group.

[0022] Figure 2A and Figure 2B Shows the images of brain microglial marker molecule IBA-1 detected by immunofluorescence and neurons observed by Golgi staining.

[0023] Figure 3 Shows the results of FITC-dextran detection of blood-brain barrier permeability and RT-PCR method detection of the expression of tight junction molecule ZO-1 and tight junction protein-5 (Claudin-5) in brain tissue (1: control group; 2: lipopolysaccharide model group; 3: lipopolysaccharide model + sodium β-hydroxybutyrate group; 4: lipopolysaccharide model + sodium β-hydroxybutyrate + vitamin D3 group; 5: lipopolysaccharide model + vitamin D3 group); *P < 0.05, relative to the control group; #P < 0.05, relative to the lipopolysaccharide model group.

[0024] Figure 4 The results of detecting intestinal barrier permeability with FITC-dextran and detecting the expression of tight junction molecule ZO-1 and connexin-5 in colon tissue by RT-PCR are shown (1: control group; 2: lipopolysaccharide model group; 3: lipopolysaccharide model + sodium β-hydroxybutyrate group; 4: lipopolysaccharide model + sodium β-hydroxybutyrate + vitamin D3 group; 5: lipopolysaccharide model + vitamin D3 group); **P < 0.01, relative to the control group; #P < 0.05, relative to the lipopolysaccharide model group.

[0025] Figure 5 The results of detecting the expression and protein content of vitamin D3 receptor (VDR) in brain tissue and colon tissue by RT-PCR and Western blotting are shown (1: control group; 2: lipopolysaccharide model group; 3: lipopolysaccharide model + sodium β-hydroxybutyrate group; 4: lipopolysaccharide model + sodium β-hydroxybutyrate + vitamin D3 group; 5: lipopolysaccharide model + vitamin D3 group); **P < 0.01 or *P < 0.05, relative to the control group; #P < 0.05, relative to the lipopolysaccharide model group. Detailed implementation manners

[0026] To make the technical problems to be solved, the technical solutions adopted and the advantages of the present invention clearer, the present invention will be described in detail below with reference to the accompanying drawings and specific embodiments. The following embodiments are used to illustrate the present invention, but are not used to limit the scope of the present invention.

[0027] Ketone bodies are generally considered to be carriers for the liver to supply energy to tissues such as the brain and heart when the supply of sugar for energy is insufficient. In addition to providing energy, β-hydroxybutyric acid, the main component of ketone bodies, can also act as a bioactive small molecule, easily pass through the blood-brain barrier, and play important neuroprotective and vascular protective roles. The ketogenic diet has long been used in the treatment of central nervous system diseases such as epilepsy and Alzheimer's disease. Compared with the ketogenic diet, supplementing β-hydroxybutyric acid is easier to adhere to, the dosage is better controlled, and it is more conducive to exerting its protective effect and avoiding adverse reactions, thus becoming a popular research method. Studies have found that β-hydroxybutyric acid has the effect of maintaining neuronal integrity and can play a neuroprotective role through multiple mechanisms such as inhibiting the activation of the NF-κB pathway, inhibiting histone deacetylase, and inhibiting the NLRP3 inflammasome. However, whether β-hydroxybutyric acid exerts a protective effect by reducing the high permeability of the blood-brain barrier and intestinal barrier and its internal molecular mechanism are currently unclear.

[0028] Vitamin D3 is a fat-soluble steroid hormone, mainly derived from dietary intake and skin synthesis. 7-Dehydrocholesterol in the skin can be converted into vitamin D3 (cholecalciferol) after activation by ultraviolet light, and then transformed into 25(OH)D3 under the action of liver 25-hydroxylase, and then converted into the highly active metabolite 1,25(OH)2D3 by renal 1α-hydroxylase. 1,25(OH)2D3 can penetrate the blood-brain barrier, and 1α-hydroxylase also exists in the cerebral cortex, indicating that 1,25(OH)2D3 can be produced in the central nervous system. Studies have found that vitamin D3 plays an important role in the brain by regulating cell differentiation, proliferation and maintaining calcium homeostasis after binding to its specific receptor, vitamin D3 receptor (VDR). VDR is widely distributed in various tissues of the body, and is abundantly distributed in vascular endothelium, neurons and glial cells. After vitamin D3 binds to VDR, it can inhibit the activation of reactive oxygen species and the activity of NF-κB, and at the same time up-regulate the expression and activity of tight junction proteins, maintaining the integrity of the blood-brain barrier and intestinal barrier, and playing an improving role in many diseases, especially neurological diseases. However, pathological stimuli such as inflammation can inhibit the expression of VDR, thereby weakening the protective effect of vitamin D3.

[0029] The inventors of the present application have found that β-hydroxybutyric acid plays a protective role by reducing the high permeability of the blood-brain barrier and intestinal barrier, as well as its underlying molecular mechanism.

[0030] The inventors of the present application have also found that β-hydroxybutyric acid can initiate the expression of the vitamin D3 specific receptor VDR, β-hydroxybutyric acid has a synergistic effect with vitamin D3, and combining β-hydroxybutyrate with vitamin D3 can better reduce the high permeability of the blood-brain barrier and intestinal barrier, and play a therapeutic role in diseases related to the high permeability of the blood-brain barrier and intestinal barrier.

[0031] It should be noted that the experimental methods used in the present invention are all conventional methods unless otherwise specified.

[0032] The reagents and materials used in the present invention are all prepared by conventional methods or obtained through commercial channels unless otherwise specified.

[0033] It should be understood that the terms "having", "comprising" and "including" used herein are synonyms and are open-ended, and do not exclude the presence or addition of other elements or their components.

[0034] As used herein, the term "therapeutically effective amount" refers to the total amount of each active component of a pharmaceutical composition or method sufficient to demonstrate a meaningful patient benefit.

[0035] As used herein, the term "pharmaceutically acceptable" refers to a moiety or substance that, within the scope of medical judgment, is suitable for use in humans or animals without causing undesirable biological effects such as undue toxicity, irritation, allergic reactions, etc.

[0036] Unless otherwise specified, the experimental instruments and the like used in the present invention are obtained commercially.

[0037] Unless otherwise specified, the parts used in the present invention are parts by weight, % is % by weight, and the temperature is in degrees Celsius.

[0038] The oral administration preparation of the pharmaceutical composition of the present invention is: sodium β-hydroxybutyrate solid for human use at 10 mg / kg / day, vitamin D3 (dissolved in sesame oil) at 0.1 μg / kg / day, taken orally. The dose converted for mouse experiments is sodium β-hydroxybutyrate aqueous solution at 300 mg / kg / day, and vitamin D3 (dissolved in corn oil to exclude the possible beneficial effects of the solvent) at 10 μg / kg / day, administered by gavage.

[0039] In the following examples, the following raw materials and methods are adopted.

[0040] Unless otherwise specified, the control group in the examples of the present application is gavaged with 0.1 mL of pure water.

[0041] The sodium β-hydroxybutyrate used in the present application is purchased from J&K Scientific Ltd. (batch number 204762), and vitamin D3 is purchased from Solarbio Science & Technology Co., Ltd. (batch number V8070).

[0042] The ratio of vitamin D3 to sesame oil is 100 μg of vitamin D3 dissolved in 0.1 mL of sesame oil.

[0043] Example 1 <Mouse Toxicity Test>

[0044] 8-week-old male ICR mice weighing 28.0 ± 2.93 g were used. They were randomly divided into a dosing group and a control group, with 8 mice in each group. The dosing group was respectively given sodium β-hydroxybutyrate at a dose of 1 g per kilogram of body weight per day, and vitamin D3 (dissolved in 0.1 mL of corn oil) at a dose of 100 μg per kilogram of body weight per day. Before the experiment, the mice were first fasted for 12 hours, and the appearance and activity of the mice were observed and recorded, and they were weighed. After each group of mice was gavaged, the toxic reactions of the mice were immediately observed, and they were continuously recorded for 14 days, including whether there were abnormalities in the facial features, limbs, behavior, spirit, appetite, urine and feces of the mice. If any mouse died, an autopsy was performed immediately for pathological examination.

[0045] The results showed that after the mice were administered the drug, their diet, activity, hair color, and feces were all normal, and no animals died. There was no significant change in the body weight of the mice in the drug administration group compared with the control group. After the experiment ended, the mice were dissected by cutting the spinal cord. No abnormalities were found in the internal organs such as the heart, liver, spleen, lungs, and kidneys, and no abnormalities or tumors were found in the tissues and organs or the abdominal cavity. It can be seen that the dosage of the drug administered by gavage to the mice was 10 times higher than the conventional dosage, and no acute toxicity reaction was observed.

[0046] Example 2 <Therapeutic Efficacy Test for Nerve Injury>

[0047] Eight-week-old male ICR mice weighing 26.50 ± 1.57 g were randomly divided into a control group, a lipopolysaccharide model group, a lipopolysaccharide model + sodium β-hydroxybutyrate group, a lipopolysaccharide model + sodium β-hydroxybutyrate + vitamin D3 group, a lipopolysaccharide model + vitamin D3 group, a sodium β-hydroxybutyrate group, and a vitamin D3 group, with 8 mice in each group. The lipopolysaccharide model group was given intraperitoneal injection of lipopolysaccharide LPS (1 mg / kg), injected every other day for a total of 7 times to establish a classical systemic inflammation animal model. Sodium β-hydroxybutyrate aqueous solution was administered by gavage at 300 mg / kg, and vitamin D3 (dissolved in corn oil) was administered by gavage at 10 μg / kg. The control group was given 0.1 mL of pure water by gavage, and the drugs were administered continuously for 18 days. On the 14th day of drug administration, open field test behavioral detection was performed, and the results were as Figure 1 shown. After the drug administration ended, the animals were sacrificed, and brain tissues were taken. Immunofluorescence was used to detect the microglial cell marker molecule IBA-1 in the brain to reflect the activation of microglial cells. At the same time, Golgi staining was used to observe the morphological changes of neurons, and the results were as Figure 2A and Figure 2B shown.

[0048] Figure 1 A in showed that there was no significant difference in body weight among the groups, indicating random grouping in the experiment. In the open field test behavioral test, compared with the control group, the time that the mice in the lipopolysaccharide model group stayed in the middle area was significantly reduced, while the time was significantly extended after intervention, and the time of the mice in the lipopolysaccharide model + sodium β-hydroxybutyrate + vitamin D3 group was extended most significantly ( Figure 1 B in ). At the same time, compared with the control group, the relative movement distance of the mice in the lipopolysaccharide model group in the middle area was significantly shortened, while the movement distance was significantly increased after intervention, and the distance of the mice in the lipopolysaccharide model + sodium β-hydroxybutyrate + vitamin D3 group increased most significantly ( Figure 1 C in ).

[0049] Figure 1 The results in showed that both sodium β-hydroxybutyrate and vitamin D3 could improve the behavioral abnormalities in mice caused by lipopolysaccharide, and the combined effect of the two was the best.

[0050] Figure 2AIt was shown that, compared with the control group, the fluorescence of microglial cells staining in the lipopolysaccharide model group was significantly increased and enhanced. After intervention, the fluorescence of microglial cells staining was significantly weakened, with the lipopolysaccharide model + sodium β-hydroxybutyrate + vitamin D3 group being the most significant. Figure 2B It was shown that, compared with the control group, the nerve fibers in the lipopolysaccharide model group were significantly entangled and shortened. After intervention, the nerve fibers were significantly increased and lengthened, with the lipopolysaccharide model + sodium β-hydroxybutyrate + vitamin D3 group being the most significant. Figure 2A and Figure 2B The results in

[0051] Example 3 <Blood-brain Barrier Permeability Study Experiment>

[0052] Eight-week-old male ICR mice with a body weight of 26.50 ± 1.57 g were used. They were randomly divided into a control group, a lipopolysaccharide model group, a lipopolysaccharide model + sodium β-hydroxybutyrate group, a lipopolysaccharide model + sodium β-hydroxybutyrate + vitamin D3 group, and a lipopolysaccharide model + vitamin D3 group, with 8 mice in each group. In the lipopolysaccharide model group, lipopolysaccharide LPS was intraperitoneally injected (1 mg / kg) every other day for a total of 7 times to establish a classical systemic inflammation animal model. Sodium β-hydroxybutyrate aqueous solution was administered by gavage at 300 mg / kg, and vitamin D3 (dissolved in corn oil) was administered by gavage at 10 μg / kg. The control group was administered 0.1 mL of pure water by gavage for 18 consecutive days. After the administration ended, the permeability of the blood-brain barrier was detected by the fluorescence probe fluorescein isothiocyanate-dextran (FITC-Dextran) method (30 minutes after injecting 10 mg of FITC-dextran dissolved in 0.1 mL into the tail vein of the mice, the animals were sacrificed, the brain was quickly removed and frozen. After frozen sectioning, it was observed under a fluorescence microscope and photographed); at the same time, the animals were sacrificed, the brain tissue was taken, and the expressions of tight junction molecule ZO-1 and tight junction protein-5 (Claudin-5) were detected by real-time PCR (Realtime-PCR, RT-PCR). The results were as Figure 3 shown.

[0053] Figure 3 A in Figure 3B in indicates that, simultaneously, compared with the control group, the expression of tight junction protein occludin-5 in the brain tissue of mice in the lipopolysaccharide model group was significantly reduced, and ZO-1 was also reduced. After intervention, the expression of occludin-5 and ZO-1 increased significantly, with the most obvious increase in the mice of the lipopolysaccharide model + sodium β-hydroxybutyrate + vitamin D3 group. The above results indicate that both sodium β-hydroxybutyrate and vitamin D3 can improve the high permeability of the blood-brain barrier induced by lipopolysaccharide, and the combined effect of the two is the best.

[0054] Example 4 <Intestinal Barrier Permeability Study Experiment>

[0055] Eight-week-old male ICR mice weighing 26.50 ± 1.57 g were randomly divided into a control group, a lipopolysaccharide model group, a lipopolysaccharide model + sodium β-hydroxybutyrate group, a lipopolysaccharide model + sodium β-hydroxybutyrate + vitamin D3 group, and a lipopolysaccharide model + vitamin D3 group, with 8 mice in each group. In the lipopolysaccharide model group, lipopolysaccharide LPS was intraperitoneally injected (1 mg / kg), every other day for a total of 7 times, to establish a classical systemic inflammation animal model. Sodium β-hydroxybutyrate aqueous solution was administered by gavage at 300 mg / kg, and vitamin D3 (dissolved in corn oil) was administered by gavage at 10 μg / kg. The control group was given 0.1 mL of pure water by gavage, and the drugs were continuously administered for 18 days. After the administration, the permeability of the intestinal barrier was detected by the FITC-dextran method (after gavage with 0.1 mL of 20 mg FITC-dextran (fluorescein isothiocyanate dextran) for 3 h, blood was taken and serum was separated, and the content of FITC-dextran in the blood was detected by a fluorescence microplate reader. The excitation wavelength and emission wavelength used were 490 nm and 530 nm, respectively); at the same time, the animals were sacrificed, colon tissues were taken, and the expression of tight junction molecules ZO-1 and occludin-5 was detected by RT-PCR. The results are as Figure 4 shown.

[0056] Figure 4 A in shows that for the detection of intestinal barrier permeability by FITC-dextran, compared with the normal group, the fluorescence intensity of FITC in the blood of the lipopolysaccharide model group was significantly enhanced. After intervention, the fluorescence intensity was significantly weakened, with the most obvious decrease in the lipopolysaccharide model + sodium β-hydroxybutyrate + vitamin D3 group. Figure 4 B in shows that, simultaneously, compared with the control group, the expression of tight junction proteins occludin-5 and ZO-1 in the intestinal tissues of mice in the lipopolysaccharide model group was significantly reduced. After intervention, the expression of both increased significantly, with the most obvious increase in the mice of the lipopolysaccharide model + sodium β-hydroxybutyrate + vitamin D3 group. The above results indicate that both sodium β-hydroxybutyrate and vitamin D3 can improve the high permeability of the intestinal barrier induced by lipopolysaccharide, and the combined effect of the two is the best.

[0057] Example 5 <Vitamin D3 Receptor (VDR) Expression Study Experiment>

[0058] Eight-week-old male ICR mice weighing 26.50 ± 1.57 g were randomly divided into a control group, a lipopolysaccharide model group, a lipopolysaccharide model + sodium β-hydroxybutyrate group, a lipopolysaccharide model + sodium β-hydroxybutyrate + vitamin D3 group, and a lipopolysaccharide model + vitamin D3 group, with 8 mice in each group. In the lipopolysaccharide model group, lipopolysaccharide (LPS) was intraperitoneally injected (1 mg / kg) every other day for a total of 7 times to establish a classical systemic inflammation animal model. Sodium β-hydroxybutyrate aqueous solution was administered by gavage at 300 mg / kg, vitamin D3 (dissolved in corn oil) was administered by gavage at 10 μg / kg, and the control group was administered 0.1 mL of pure water by gavage for 18 consecutive days. After the administration ended, the animals were sacrificed, and brain tissues and colon tissues were taken. The expression and protein content of VDR were detected by RT-PCR and Western blotting. The results are as Figure 5 shown.

[0059] The expression and protein content of brain VDR ( Figure 5 A and B in Figure 5 ) showed that compared with the normal group, the expression and protein content of VDR in the lipopolysaccharide model group were significantly reduced, while after the intervention with sodium β-hydroxybutyrate group and sodium β-hydroxybutyrate + vitamin D3 combination, the expression and protein content of VDR were significantly increased. The expression and protein content of intestinal VDR ( Figure 5 C and D in Figure 5 ) showed that compared with the normal group, the expression and protein content of VDR in the lipopolysaccharide model group were significantly reduced, while after the intervention, the expression and protein content of VDR were significantly increased, and the increase was most obvious with the intervention of sodium β-hydroxybutyrate and sodium β-hydroxybutyrate + vitamin D3 combination. The above results indicate that sodium β-hydroxybutyrate can reverse the decrease in the expression and protein content of VDR in the brain and intestine caused by inflammatory stimulation.

[0060] Example 6 <Typical Cases>

[0061] Case 1, a male patient, 9 years old, with autism and allergic asthma for 3 years. Symptoms: Hyperactivity, sleep disorder, poor social and language abilities, and frequent asthma attacks. He took a combined drug (food-grade sodium β-hydroxybutyrate solid at 10 mg / kg / day, vitamin D3 at 0.1 μg / kg / day) for 3 consecutive years. Hyperactivity basically disappeared, sleep was basically normal, social and language abilities were significantly improved, and asthma did not attack.

[0062] Case 2, a female patient, 40 years old, with allergic asthma and general weakness for 10 years. Symptoms: Allergic asthma, fatigue, fear of cold, and difficulty in concentrating. She took a combined drug (food-grade sodium β-hydroxybutyrate solid at 10 mg / kg / day, vitamin D3 at 0.1 μg / kg / day) for 3 consecutive years. Allergic asthma basically disappeared, physical fitness was enhanced, concentration was improved, and the mind was clear.

[0063] The number of devices and the processing scale described here are used to simplify the description of the present invention. Applications, modifications, and variations of the present invention will be apparent to those skilled in the art.

[0064] Although the embodiments of the present invention have been disclosed above, it is not limited to the applications listed in the specification and the embodiments. It can be fully applied to various fields suitable for the present invention. For those familiar with the art, additional modifications can be easily achieved. Therefore, without departing from the general concept defined by the claims and the equivalent scope, the present invention is not limited to the specific details shown and described herein.

Claims

1. Use of a pharmaceutical composition in the preparation of a drug for treating diseases associated with high permeability of the blood-brain barrier and intestinal barrier, characterized in that, The composition contains a pharmaceutically acceptable β-hydroxybutyrate and vitamin D as active ingredients. Among them, the diseases related to the high permeability of the blood-brain barrier and intestinal barrier are: neurological diseases and allergic diseases, and among them, the neurological diseases are autism, depression, anxiety and Parkinson's syndrome.

2. The use according to claim 1, characterized in that, In the pharmaceutical composition, the content of β-hydroxybutyrate is 8,000-30,000 parts by weight, and the content of vitamin D is 0.08-1 part by weight.

3. The use according to claim 2, characterized in that, In the pharmaceutical composition, the amount of β-hydroxybutyrate is 8,000-20,000 parts by weight, and the amount of vitamin D is 0.08-0.5 part by weight.

4. The use according to claim 2, characterized in that, In the pharmaceutical composition, the amount of β-hydroxybutyrate is 10,000-15,000 parts by weight, and the amount of vitamin D is 0.1-0.5 part by weight.

5. The use according to claim 2, characterized in that, In the pharmaceutical composition, the amount of β-hydroxybutyrate is 10,000 parts by weight, and the amount of vitamin D is 0.1 part by weight.

6. The use according to any one of claims 1 to 5, characterized in that, The β-hydroxybutyrate includes the sodium salt, calcium salt or magnesium salt of pharmaceutically acceptable β-hydroxybutyric acid.

7. Use according to any one of claims 1 to 5, characterized in that, The vitamin D includes: vitamin D3, vitamin D2, alfacalcidol, calcifediol (25-hydroxyvitamin D), calcitriol (1α,25-dihydroxyvitamin D), dihydrotachysterol (DHT) or a combination thereof.

8. The use according to claim 7, characterized in that, The vitamin D is active vitamin D3.

9. The use according to any one of claims 1 to 5, characterized in that, The pharmaceutical composition further contains a pharmaceutically acceptable solvent.

10. The use according to claim 9, wherein, The pharmaceutically acceptable solvent is selected from one or more of sesame oil, olive oil, soybean oil, corn oil, grape seed oil, evening primrose oil and sunflower oil.

11. The use according to any one of claims 1 to 5, characterized in that, The pharmaceutical composition further contains a pharmaceutically acceptable excipient.

12. The use according to claim 11, characterized in that, The pharmaceutically acceptable excipient contains lactose and starch as fillers; magnesium stearate and calcium stearate as lubricants; plasticizers, disintegrants and preservatives.

13. The use according to any one of claims 1 to 5, characterized in that, The pharmaceutical composition is made into a pharmaceutically acceptable dosage form; the dosage form includes tablets, capsules, granules, disintegrants, dispersants, pills, oral liquids and films.

14. The use according to any one of claims 1 to 5, characterized in that, The allergic disease is allergic asthma.

15. The use according to any one of claims 1 to 5, characterized in that, The neurological disease is autism.

16. The use according to any one of claims 1 to 5, characterized in that, The use is to reduce the high permeability of the blood-brain barrier and intestinal barrier caused by systemic inflammation, thereby alleviating nerve and systemic damage.

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