Application of adenosine receptor inhibitor in preparation of medicine for treating hyperlipidemia and cardiovascular and cerebrovascular diseases

Abstract

The invention provides application of an adenosine receptor inhibitor in preparation of a medicine for treating hyperlipidemia and cardiovascular and cerebrovascular diseases. The GPR146 is a GPCR orphan receptor which is found in the earlier stage and used for regulating blood lipid metabolism. In a process of screening an endogenous ligand of GPR146, we accidentally find that GPR146 can form a functional complex with another GPCR family member A2AR to jointly respond to activation of adenosine. The adenosine inhibitor can achieve a remarkable blood fat reducing effect, a key theoretical basis is laid for developing the blood fat reducing medicine targeting GPCR, and the adenosine inhibitor can be widely applied to treatment of hyperlipidemia and related cardiovascular and cerebrovascular diseases.

Description

Technical Field

[0001] The present invention relates to the field of biomedical technology, and in particular to the use of adenosine receptor inhibitors in the preparation of drugs for treating hyperlipidemia and cardiovascular and cerebrovascular diseases. Background Art

[0002] Hyperlipidemia, especially hypercholesterolemia, is an important cause of a series of metabolic diseases such as atherosclerosis, coronary heart disease, fatty liver, and type 2 diabetes. Studies in recent years have also found that hyperlipidemia also plays an important role in neurodegenerative diseases. Currently available small molecule lipid-lowering drugs mainly target cholesterol synthesis and absorption pathways, but these drugs have limited cholesterol-lowering effects and have certain side effects. Inhibitor drugs targeting secretory proteins PCSK9 and ANGPTL3 that have been launched in recent years have a good lipid-lowering effect, but these drugs are expensive and require long-term injections (Table 1). Therefore, cheap, convenient and efficient small molecule lipid-lowering drugs have always been an important direction of research and development for major pharmaceutical companies (Chen LG, et a l. SciChina Life Sci. 2019).

[0003] Table 1: Major cholesterol-lowering drugs on the market

[0004]

[0005] G protein-coupled receptors (GPCRs) are a type of membrane protein receptor with seven transmembrane domains that transmit extracellular signals into cells by interacting with intracellular G proteins. The human genome encodes thousands of GPCR family members, which participate in all aspects of the body and cell life activities. It is particularly important that more than one-third of the small molecule drugs currently on the market target the GPCR family, which also plays an important role in drug development and disease treatment.

[0006] Hyperlipidemia has significant genetic characteristics. The applicant previously discovered through genetic analysis and functional experiments of a population cohort that the GPCR orphan receptor GPR146 can respond to the activation of serum contents, and inhibiting its expression significantly reduces serum cholesterol levels, without other side effects (F. Han et al., Hypercholesterolemia risk-associated GPR146 is an orphan G-protein coupled receptor that regulates blood cholesterol levels in humans and mice. Cell Res 30, 363-365 (2020)). Our results were also reported by Dr. Cowan's laboratory at Harvard Medical School at almost the same time (H. Yu et al., GPR146 Deficiency Protects against Hypercholesterolemia and Atherosclerosis. Cell, 1276-1288e1214 (2019)). GPR146 is also the first GPCR reported to directly regulate serum cholesterol levels. It has become a new important target for the development of lipid-lowering drugs and has received widespread attention (Rom O, et al, Curr Opin Lipidol, 2021). At present, the structure of GPR146 is still unclear, and its ligands and related agonists and inhibitors have not been reported, which has become the main bottleneck for the development of GPR146 drugs.

[0007] Therefore, it is necessary to develop a drug targeting GPR146 for the treatment of hyperlipidemia and cardiovascular and cerebrovascular diseases. Summary of the invention

[0008] The purpose of the present invention is to provide the use of adenosine receptor inhibitors in the preparation of drugs for the treatment of hyperlipidemia and cardiovascular and cerebrovascular diseases. The present application finds that adenosine inhibitors have a significant lipid-lowering effect and will have important application prospects in lowering blood lipids and treating cardiovascular diseases.

[0009] To achieve the above object, the present invention adopts the following technical solution:

[0010] In a first aspect of the present invention, there is provided use of an adenosine receptor inhibitor in the preparation of a medicament for treating hyperlipidemia and cardiovascular and cerebrovascular diseases.

[0011] Further, the adenosine receptor inhibitor includes at least one of Istradefylline (KW6002), CSC, SCH 58261, SCH 442416, ZM241,385, VER 6947, VER 7835, Schering compound, preladenant (SCH420814), and SYN115, specifically Figure 1 shown.

[0012] Furthermore, the medicine also includes pharmaceutically acceptable excipients and carriers.

[0013] Furthermore, the auxiliary material includes at least one of a filler, a disintegrant, a binder, an excipient, a diluent, a lubricant, a sweetener or a colorant.

[0014] Furthermore, the dosage form of the drug includes at least one of granules, tablets, pills, capsules, injections or dispersions.

[0015] Furthermore, the medicine for treating hyperlipidemia and cardiovascular and cerebrovascular diseases includes a medicine for lowering cholesterol.

[0016] In the second aspect of the present invention, there is provided the use of an inhibitor of the complex of GPR146 and adenosine receptor A2AR in the preparation of a drug for treating hyperlipidemia and cardiovascular and cerebrovascular diseases.

[0017] One or more technical solutions in the embodiments of the present invention have at least the following technical effects or advantages:

[0018] The present invention provides an application of adenosine receptor inhibitors in the preparation of drugs for treating hyperlipidemia and cardiovascular and cerebrovascular diseases. In screening the endogenous ligand of the cholesterol regulatory receptor GPR146, we unexpectedly found that GPR146 can form a functional complex with A2AR and jointly respond to the activation of adenosine, indicating that the adenosine receptor A2AR is a key receptor for regulating cholesterol metabolism. At the same time, the inventors of the present application found that adenosine receptor inhibitors have a significant lipid-lowering effect. This shows that adenosine receptor inhibitors will have important application prospects in the preparation of drugs for lowering blood lipids and treating cardiovascular diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the following briefly introduces the drawings required for use in the description of the embodiments. Obviously, the drawings described below are some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without creative work.

[0020] Figure 1 It is the structural formula of various adenosine receptor inhibitors.

[0021] Figure 2 Adenosine specifically activates the downstream signaling pathway of GPR146. Left: Adenosine can dose-dependently activate GPR146 and its downstream cAMP-PKA-CRE signaling pathway; Right: Gene silencing of GPR146 inhibits the activation effect of adenosine. A: Adenosine.

[0022] Figure 3 Among nucleoside substances, only adenosine can activate GPR146 and its downstream cAMP-PKA-CRE signaling pathway.

[0023] Figure 4 The activation effect of adenosine depends on A2AR. Left: Gene silencing of A2AR inhibits the activation effect of adenosine; Right: A2AR-specific inhibitors inhibit the activation effect of adenosine. A: Adenosine;

[0024] Figure 5 A2AR specifically interacts with GPR146. IP: immunoprecipitation;

[0025] Figure 6 Gene knockout of liver Gpr146 significantly reduced serum cholesterol levels in mice. NC: normal chow; HFHC: high-fat, high-cholesterol chow.

[0026] Figure 7 KW6002, an A2AR-specific inhibitor, significantly reduced serum cholesterol levels in mice.

[0027] Figure 8 Doxycycline induced the expression of GPR146 protein. GPR146 and internal reference Vinculin were detected by immunoblotting using their specific antibodies.

[0028] Fig. 9 This is the plasmid map of the TET ON lentiviral expression vector. DETAILED DESCRIPTION

[0029] The present invention will be described in detail below in conjunction with specific implementations and examples, and the advantages and various effects of the present invention will be more clearly presented. It should be understood by those skilled in the art that these specific implementations and examples are used to illustrate the present invention, rather than to limit the present invention.

[0030] Throughout the specification, unless otherwise specifically stated, the terms used herein should be understood as meanings commonly used in the art. Therefore, unless otherwise defined, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art to which the present invention belongs. In the event of a conflict, the present specification takes precedence.

[0031] Unless otherwise specified, various raw materials, reagents, instruments and equipment used in the present invention can be purchased from the market or obtained by existing methods.

[0032] The technical solution of the embodiment of the present application is to solve the above technical problems, and the overall idea is as follows:

[0033] GPR146 is able to respond to activation by serum contents, and inhibition of its expression significantly reduces serum cholesterol levels.

[0034] The inventors of this application first discovered that adenosine activated the cAMP-PKA signaling pathway dependent on GPR146. In screening the endogenous ligands of GPR146, we unexpectedly found that GPR146 can form a functional complex with A2AR and respond to the activation of adenosine together, indicating that the adenosine receptor A2AR is the key receptor for regulating cholesterol metabolism. Adenosine can activate the cAMP-PKA signaling pathway through adenosine-A2AR. 2A The R pathway regulates blood lipid levels. Therefore, it can be inferred that adenosine receptor inhibitors can be used to prepare drugs that lower cholesterol.

[0035] Adenosine in the extracellular space binds to adenosine receptors on the plasma membrane, inducing conformational changes in the receptors and activating G proteins coupled downstream of the receptors and their downstream signal transduction pathways to exert physiological functions. The adenosine receptor family is currently divided into four main receptors: A1R, A2R, A3R, A4R, A5R, A6R, A7R, A8R, A9R, A10R, A21R, A11R, A12R, A13R, A14R, A15R, A16R, A17R, A18R, A19R, A20R, A21R, A31R, A41R, A51R, A19R, A20R, A31R, A41R, A5 2A R.A. 2B R, A3R, among which A1R, A 2A A R and A3R receptors have a higher affinity for adenosine; A 2B R is a low-affinity receptor that requires a high concentration of adenosine to activate A 2B R. A1R and A3R and G protein subtype G i and G o Coupling, inhibiting the activity of adenylate cyclase and reducing the intracellular cAMP concentration; A 2A R and A 2B G protein G s The cytokine couples with the mitochondria to activate adenylate cyclase, thereby increasing the concentration of cyclic adenosine monophosphate (cAMP) and activating downstream protein kinase A (PKA). 2A R is mainly expressed in the liver, heart, lungs, brown adipose tissue, immune system (spleen, thymus, immune cells and platelets) and central nervous system.

[0036] Adenosine-A 2A The Adenosine-A R pathway has been found to play an important role in many diseases in the human body, such as cancer and Parkinson's disease. 2AThe A R pathway has been found to play an important role in many diseases in the human body, such as cancer and Parkinson's disease. 2A Adenosine receptor A R antagonists have become an attractive research area. 2A Adenosine receptor antagonists can be divided into two main categories: xanthine derivatives and azapolycyclic systems. Natural xanthine binds to all adenosine receptors at micromolar concentrations (15-80 μM) without any selectivity. The researchers randomly replaced the ring groups of xanthine and screened out Adenosine receptor antagonists. 2A R has high selectivity, while other adenosine receptors have low affinity for small molecules, such as DMPX, KF17837, etc. However, due to the poor water solubility and photoisomerism of xanthine derivatives, small molecule compounds with nitrogen-polycyclic structures have been developed, which have a strong affinity for A 2A R also has specific affinity, such as SCH 58261, SCH 63390, ZM241385, etc. At present, a variety of A2AR antagonists including Istradefylline (KW6002), CSC, SCH 58261, SCH 442416, ZM241,385, VER6947, VER 7835, Schering compound, preladenant (SCH 420814), SYN115, etc. have entered clinical trials and are used in the treatment of cancer immunotherapy, nervous system diseases, etc.

[0037] The full name of KW6002 is (E) 1,3-diethyl-8-(3,4-dimethoxyphenylvinyl)-7-methylxanthine, which belongs to the class of phenylvinylxanthine compounds. KW6002 is highly selective for A2AR: its affinity to A2AR is 60 times that of A1R in rats and 800 times that of A1R in humans. The equilibrium dissociation constant (Ki value) of KW6002 and A2AR is 2.2nM. The elimination half-life of KW6002 in the human body is about 47h, which is the longest half-life among the currently available A2A receptor antagonists. It can easily cross the blood-brain barrier after oral administration. KW6002 has shown positive effects in the treatment of Parkinson's disease. Phase III clinical studies are being conducted in many parts of the world, and it has been approved for clinical use in Japan.

[0038] Therefore, the inventors of this application selected KW6002 as a representative adenosine receptor inhibitor for research. We found that feeding KW6002 can significantly reduce serum cholesterol levels ( Figure 8Left). Cholesterol in serum is mainly divided into low-density lipoprotein cholesterol (LDL) and high-density lipoprotein cholesterol (HDL). LDL is an important cause of cardiovascular and cerebrovascular diseases such as atherosclerosis, while HDL is considered to have a protective effect on the cardiovascular system. We found that KW6002 mainly reduced the level of LDL cholesterol and had no significant effect on HDL cholesterol ( Figure 8 Right), indicating that KW6002 has important application prospects in the treatment of hypercholesterolemia and cardiovascular and cerebrovascular diseases.

[0039] The application of the adenosine receptor inhibitor of the present application in the preparation of drugs for treating hyperlipidemia and cardiovascular and cerebrovascular diseases will be described in detail below in combination with examples and experimental data.

[0040] Example 1: GPR146 and A2AR form a functional complex and jointly respond to adenosine activation

[0041] 1. Adenosine activation of the cAMP-PKA signaling pathway depends on GPR146

[0042] In previous work, we found that the downstream signaling pathway regulated by GPR146 is cAMP-PKA-CRE (F.Han et al., Hypercholesterolemia risk-associated GPR146 is an orphan G-protein coupled receptor that regulates blood cholesterol levels in humans and mice. Cell Res 30, 363-365 (2020).). To find the endogenous ligand of GPR146, we simultaneously expressed GPR146 and the CRE luciferase reporter system in HEK293T cells. The specific steps are as follows:

[0043] 1. Establishment of GPR146 TET ON inducible expression system

[0044] GPR146 has a seven-transmembrane structure, which can easily lead to folding errors when overexpressed. Rho guide peptide can effectively assist the folding and accurate intracellular localization of GPCR family proteins and is widely used in GPCR research. In this study, we overexpressed GPR146 by constructing a Rho-GPR146 fusion expression plasmid. The TET ON inducible expression system can accurately control the amount of protein expression, so we chose this system to conduct GPR146 overexpression research. Using standard molecular cloning technology, we inserted the Rho-GPR146 gene fragment (shown in SEQ ID NO.1) into the TET ON lentiviral expression vector (gift from, map as shown in Fig. 9 As shown in Figure 2, the original sequence of the TET ON lentiviral expression vector is shown in SEQ ID NO.2) in the FseI and AgeI sites. It was then co-transfected with the auxiliary plasmids PSPAX and PMD2G and packaged into lentiviral particles in 293T cells. We then used the virus to infect HEK293 cells and screened the blasticidin resistance gene carried by the expression vector to obtain a TET ON inducible expression cell line that stably expressed Rho-GPR146. Figure 2 As shown, different concentrations of doxycycline (abbreviated as Dox) can induce the expression of Rho-GPR146 in a dose-dependent manner, indicating that the cell line was successfully constructed. All subsequent experiments used 2 μg / ml doxycycline for induction of expression.

[0045] 2. Screening of GPR146 ligands using the CRE-Luciferase reporter system

[0046] On the first day, the Rho-GPR146 stable expression cell line with good growth status was subcultured, and the cell density was set at 4×10 6 / 10cm dish.

[0047] On the second day, when the cells grew to 60%-80% density, luciferase reporter plasmid transfection was performed using liposomes. The transfected plasmids included CRE-Luciferase reporter plasmid (Promega) and hRenilla internal reference plasmid (Promega). CRE is a regulatory element that responds to the transcription factor Creb and is widely used in the study of cAMP-PKA-Creb signaling pathway activity.

[0048] On the third day, 24 h after transfection, the cells were trypsinized and plated into 96-well plates, and doxycycline was added to induce the expression of GPR146;

[0049] On the fourth day, the cell culture medium was changed to DMEM containing 0.1% BSA.

[0050] On the fifth day, different concentrations of adenosine, guanosine, thymidine, uridine, cytidine and inosine were added, and luciferase activity was detected after 6 hours.

[0051] By screening more than 1,000 metabolite small molecules, we found that adenosine can specifically activate GPR146 activity and its downstream cAMP-PKA-Cre signaling pathway ( Figure 3 Left), and gene silencing of GPR146 can block this activation effect, indicating that the activation effect of adenosine is GPR146-dependent ( Figure 3Right). The siRNA sequences used for gene silencing were predicted by the siRNA gene silencing sequence prediction software on the Merck official website, and the three siRNA sequences with the highest scores were selected for synthesis. The siRNA synthesis was completed by Shanghai Jima. The synthesized siRNA was directly transfected with cells using Lipofectamine RNAiMAX (Thermofilter, Cat. No. 13778030), and the specific transfection process was carried out in full accordance with the official instructions. Two of the siRNAs with good gene silencing effects were used in the current experiment. The siRNA sequences are shown in Table 2.

[0052] Adenosine, guanosine, thymidine, uridine, cytidine, and inosine are all nucleotide metabolites. We found that among these nucleoside small molecules, only adenosine can activate GPR146, indicating that adenosine is a specific small molecule that activates GPR146 ( Figure 4 ).

[0053] Table 2

[0054]

[0055] 2. Adenosine activation of the cAMP-PKA signaling pathway depends on A2AR

[0056] A2AR is a known adenosine receptor (JF Chen, HK Eltzschig, BB Fredholm, Adenosine receptors as drug targets--what are the challenges Nat Rev Drug Discov 12, 265-286 (2013).). After discovering that GPR146 specifically responds to adenosine stimulation, we were prompted to verify whether A2AR plays a role in this activation process.

[0057] 1. Gene silencing of A2AR inhibits adenosine's ability to activate GPR146

[0058] (a) First, we used the shRNA gene silencing sequence prediction software on the Merck official website to predict and select the three shRNA sequences with the highest scores. The shRNA target sequence was synthesized by Beijing Qingke Biotechnology Co., Ltd. During the synthesis, the shRNA sequence contained AgeI and EcoRI restriction sites at both ends. Subsequently, it was cloned into the lentiviral expression vector pLKO.1-TRC (from Addgene, catalog number 179360) through the AgeI and EcoRI restriction sites to construct the pLKO.1-shRNA plasmid; the lentivirus was packaged in 293T cells by the same method as in Example 1.

[0059] (b) Verification of the gene silencing efficiency of the three shRNAs in 293T cells. The specific implementation is as follows: on the first day, 293T cells were plated in a six-well plate; on the second day, lentivirus infection was performed; on the third day, fresh culture medium was replaced for the cells according to the cell state; on the fourth day, the cells were harvested and the A2AR gene silencing efficiency was detected by fluorescence quantitative PCR.

[0060] Two shRNAs with good gene silencing efficiency were selected for subsequent experiments and named sh-A2AR-1 and shA2AR-2 (sequences shown in Table 3).

[0061] Table 3

[0062]

[0063] (c) In the cell lines induced by TET ON to express GPR146, A2AR gene silencing experiments were performed respectively, and the specific experimental steps were as shown in b). The transfection of luciferase reporter plasmid and the treatment conditions of adenosine were the same as those in Example 1.

[0064] 2. A2AR inhibitor KW6002 inhibits adenosine's ability to activate GPR146

[0065] Referring to Example 1, the activation ability of adenosine on GPR146 was simultaneously tested in the presence or absence of KW6002 (1 μM) in the TET ON-induced GPR146 expression cell line. Figure 5 As shown on the right, KW6002 can significantly inhibit the activation ability of adenosine on GPR146, indicating that the activation of GPR146 by adenosine depends on A2AR, and A2AR inhibitors can inhibit the activity of GPR146.

[0066] 3. A2AR forms a complex with GPR146

[0067] On the first day, 293T cells with good growth status were passaged and the density was set at 2.1×10 6 / 10cm dish.

[0068] On the second day, when the cell density reached 60%-80%, the corresponding expression plasmid was transfected. The transfection reagent was LPEI, and the ratio of LPEI to plasmid was 2μl:1μg. After 8 hours, fresh culture medium was replaced.

[0069] On the fourth day, 48 hours after transfection, cells were collected and cell lysates were prepared. Part of the cell lysate was retained as input, and part of the lysate was used for immunoprecipitation (using Anti-Flag M2 agarose gel beads). Input and immunoprecipitated samples were analyzed by immunoblotting to determine changes in the levels of GPR146 and A2AR. Using biochemical methods such as co-immunoprecipitation, we found that GPR146 can interact with A2AR ( Figure 6 ). Therefore, the inhibitor of the complex of GPR146 and adenosine receptor A2AR can be used in the preparation of drugs for treating hyperlipidemia and cardiovascular and cerebrovascular diseases. The inhibitor can inhibit GPR146 or adenosine receptor A2AR alone, or can inhibit GPR146 and adenosine receptor A2AR simultaneously.

[0070] 4. Gene knockout of Gpr146 reduces serum cholesterol levels in mice

[0071] In previous studies, we found that GPR146 gene mutations in the human population are significantly associated with hypercholesterolemia. By preparing tissue-specific gene knockout mice, we found that liver Gpr146 is an important receptor for regulating serum cholesterol. Liver-specific gpr146 gene knockout mice (Li-gpr146- / -) have lower serum cholesterol levels ( Figure 7 ), and resist hypercholesterolemia caused by a high-fat, high-cholesterol diet. The specific experimental process is as follows:

[0072] Liver-specific gpr146 knockout mice were prepared using the Cre-LoxP system. First, the F lox site was inserted into both sides of the gpr146 gene using gene targeting technology. The F lox / F lox mice were then mated with liver-specific Cre-expressing mice (Albumin-Cre), and Cre-LoxP was used to obtain liver-specific gpr146 knockout mice (Li-gpr146 - / - ). In all experiments, F lox / F lox mice born from the same litter without Cre expression served as control mice. All genotypes were verified by PCR gene amplification analysis. When mice became adults, we induced Li-gpr146 - / - The mice in the control group were fed a normal diet or fed a high-fat, high-cholesterol diet for 2 weeks, and the total cholesterol levels in the serum of the mice were further tested after overnight starvation.

[0073] Example 2: Adenosine receptor inhibitors reduce serum cholesterol levels in mice

[0074] 1. Preparation of hypercholesterolemia mouse model

[0075] The cholesterol level of wild-type mice, especially low-density lipoprotein cholesterol (LDL-C), is much lower than that of humans. In order to better simulate the serum LDL-C level of humans, we used adeno-associated virus to overexpress the LDL receptor blocker PCSK9 in the mouse liver, thereby increasing the serum LDL-C level. This method has also been widely used in mouse cholesterol metabolism research (J. Keeter WC, Carter NM, Nadler JL, Galkina EV., The AAV-PCSK9 murine model of atherosclerosis and metabolic dysfunction. Eur Heart J Open. 2022Apr 20; 2(3): oeac028.doi:10.1093 / ehjopen / oeac028). The specific method is as follows:

[0076] Step 1: Preparation of PCSK9-expressing adeno-associated virus

[0077] Adeno-associated virus is a mature method for in vivo gene delivery and has been used in clinical gene therapy. In this protocol, the PCSK9 gene fragment was first cloned into an adeno-associated virus expression vector using a standard molecular cloning method, and then co-expressed with an auxiliary plasmid (all plasmids are from Addgene), and then isolated and purified to obtain adeno-associated virus particles. The specific experimental process can be seen in (F. Han et al., Hypercholesterolemia risk-associated GPR146 is anorphan G-protein coupled receptor that regulates blood cholesterol levels in humans and mice. Cell Res30, 363-365 (2020)).

[0078] Step 2: Use PCSK9 adeno-associated virus to prepare a high cholesterol mouse model

[0079] The purified PCSK9 adeno-associated virus particles were injected into mice via tail vein (5*10 11 Particles / mouse), and 14 days later, blood was collected from the tail vein to test serum cholesterol levels, and further in vivo experiments of KW6002 were carried out.

[0080] 2. In vivo experiments of A2AR inhibitor KW6002

[0081] The high cholesterol mouse model prepared above was randomly divided into two groups. KW6002 was dissolved in corn oil. One group was gavaged with corn oil blank, and the other group was gavaged with A2AR inhibitor KW6002 (10 mg / kg). The gavage was performed once a day. All gavage operations were performed under anesthesia to reduce interference with the mice. After four consecutive days of gavage, the mice were starved for 4 hours and then anesthetized and killed. Blood was collected from the eyeballs to analyze the serum total cholesterol level. At the same time, the changes in cholesterol content in different lipoprotein components were analyzed by high-performance liquid protein chromatography (FPLC).

[0082] The results are as follows Figure 8 As shown in the results, oral administration of KW6002 significantly reduced the total cholesterol level in serum. Cholesterol in serum is mainly divided into low-density lipoprotein cholesterol and high-density lipoprotein cholesterol, of which low-density lipoprotein cholesterol is the main risk factor for cardiovascular and cerebrovascular diseases such as atherosclerosis. We found that feeding KW6002 mainly reduced low-density lipoprotein cholesterol ( Figure 8 Right), had no significant effect on high-density lipoprotein cholesterol, indicating that KW6002 can be used to treat hypercholesterolemia and cardiovascular and cerebrovascular diseases.

[0083] Finally, it should be noted that the terms "comprises," "includes," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that includes a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements that are inherent to such process, method, article, or apparatus.

[0084] Although the preferred embodiments of the present invention have been described, those skilled in the art may make other changes and modifications to these embodiments once they have learned the basic creative concept. Therefore, the appended claims are intended to be interpreted as including the preferred embodiments and all changes and modifications that fall within the scope of the present invention.

[0085] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.

Claims

1. Application of adenosine receptor inhibitors in the preparation of drugs for the treatment of hyperlipidemia and cardiovascular and cerebrovascular diseases.

2. The use according to claim 1, characterized in that: The adenosine receptor inhibitor includes at least one of Istradefylline (KW6002), CSC, SCH 58261, SCH 442416, ZM241,385, VER 6947, VER 7835, Schering compound, preladenant (SCH 420814), and SYN115.

3. The use according to claim 1, characterized in that: The medicine also includes pharmaceutically acceptable excipients and carriers.

4. The use according to claim 3, characterized in that: The auxiliary materials include at least one of a filler, a disintegrant, a binder, an excipient, a diluent, a lubricant, a sweetener and a colorant.

5. The use according to claim 1, characterized in that: The dosage form of the drug includes at least one of granules, tablets, pills, capsules, injections or dispersions.

6. The use according to claim 1, characterized in that: The medicine for treating hyperlipidemia and cardiovascular and cerebrovascular diseases includes medicine for lowering cholesterol.

7. Use of inhibitors of the complex of GPR146 and adenosine receptor A2AR in the preparation of drugs for treating hyperlipidemia and cardiovascular and cerebrovascular diseases.

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