Application of cannabidiol and allopurinol combination in preparation of medicine for preventing and treating hyperuricemia and reducing organ injury

Abstract

The invention belongs to the technical field of biological medicine, and particularly relates to application of cannabidiol and allopurinol in preparation of medicine for preventing and treating hyperuricemia and reducing organ injury. Researches find that when cannabidiol and allopurinol are combined, the effect of synergistically reducing the serum uric acid level and the serum creatinine level can be achieved, and then the effect of synergistically improving the uric acid reduction level or preventing and treating hyperuricemia is achieved; meanwhile, when cannabidiol and allopurinol are combined, the effect of synergistically reducing organ injury caused by hyperuricemia can be achieved.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine, and more specifically, relates to the use of cannabidiol and allopurinol in combination for preparing a medicament for preventing and treating hyperuricemia and reducing organ damage. Background Art

[0002] Gout is a disease caused by excessive precipitation of urate in the body at joints or non-joint sites, and the typical clinical manifestation is acute inflammatory arthritis with severe pain. The disease progression of gout is closely related to hyperuricemia, which is the basic pathological condition for the progression of gout. There is research evidence indicating that the incidence of gout is positively correlated with the serum uric acid concentration. At the same time, long-term hyperuricemia may lead to hyperuricemic nephropathy (HN).

[0003] Therefore, early detection and intervention for hyperuricemia are helpful for preventing the occurrence of gout and hyperuricemic nephropathy. The clinical definition of hyperuricemia (HUA) is based on the concentration of uric acid in the serum, and the critical value of serum uric acid is 420 μmol / L. Currently, the pathogenesis of hyperuricemia is mainly summarized into two aspects: excessive uric acid production and reduced uric acid excretion. Excessive uric acid production is mainly related to excessive purine intake in the diet or disease factors (such as diabetes, obesity, etc.); while reduced uric acid excretion is mainly caused by genetic factors and kidney function.

[0004] The existing clinically used drugs for reducing uric acid mainly include xanthine oxidase (XOD) inhibitors that reduce uric acid production and urate anion transporter 1 (URAT1) inhibitors that promote uric acid excretion in the kidneys. Among them, allopurinol, as the first-generation XOD competitive inhibitor, inhibits XOD to prevent the conversion of hypoxanthine and xanthine into uric acid, thereby reducing the uric acid levels in the blood and urine. Allopurinol is mainly used for treating primary and secondary hyperuricemia, especially hyperuricemia caused by excessive uric acid production; recurrent or chronic gout; tophus; uric acid kidney stones and / or uric acid nephropathy; and hyperuricemia with renal insufficiency. However, the long-term clinical use of allopurinol can cause organ damage, especially kidney damage and liver damage. Allopurinol can cause acute liver injury, and the hepatotoxicity of allopurinol has a relatively high fatality rate, which can cause acute liver failure or other complications with allergic symptoms; in addition, the long-term clinical use of allopurinol can also lead to allergy or kidney toxicity, thereby causing kidney damage. Summary of the Invention

[0005] Aiming at the above existing technical problems, the purpose of the present invention is to provide the use of cannabidiol and allopurinol in combination for preparing a medicament for reducing uric acid and / or treating hyperuricemia.

[0006] Another object of the present invention is to provide the use of the combination of cannabidiol and allopurinol in the preparation of a drug for reducing organ damage caused by hyperuricemia.

[0007] To achieve the above object, the present invention is realized by the following technical solutions:

[0008] The present invention claims the use of the combination of cannabidiol and allopurinol in the preparation of a drug for reducing uric acid and / or treating hyperuricemia.

[0009] The present invention conducts research from two aspects: in vitro and in vivo. In vitro, HT-29 cells are used to simulate intestinal epithelial cells in a hyperuricemic environment to study the regulatory effect of cannabidiol on ABCG2 in vitro; in vivo, a hyperuricemia mouse model is established by intraperitoneal injection of potassium oxonate and hypoxanthine to study the regulatory effect of cannabidiol on ABCG2 and URAT1 in mice, as well as its uric acid-lowering activity and kidney protection effect. The inventors found that cannabidiol can effectively regulate ABCG2 and URAT1, thereby promoting uric acid excretion to achieve a uric acid-lowering effect, and can be used to treat diseases related to the activity or expression level of ABCG2 and URAT1.

[0010] Furthermore, the inventors found through research that when cannabidiol and allopurinol are used in combination, they can synergistically improve the effect of reducing uric acid or treating hyperuricemia. The present invention uses the combination of cannabidiol and allopurinol to synergistically exert a synergistic effect of enhancing efficacy and reducing toxicity, providing a new idea for the design, synthesis, and development of new uric acid-lowering drugs.

[0011] In some embodiments, the reduction of uric acid refers to the reduction of uric acid in serum.

[0012] In some embodiments, the reduction of uric acid in serum refers to promoting the expression of ABCG2 and / or downregulating the expression of URAT1 to thereby reduce uric acid.

[0013] In some embodiments, the hyperuricemia includes gout or gout complications caused by hyperuricemia.

[0014] In some embodiments, the gout complications are one or more of gouty arthritis and urinary calculi.

[0015] In some embodiments, gout can be acute gout or chronic gout.

[0016] In some embodiments, the hyperuricemia is hyperuricemia caused by potassium oxonate and hypoxanthine.

[0017] In some embodiments, the mass ratio of cannabidiol to allopurinol is 0.8 - 5:1. Preferably, the mass ratio of cannabidiol to allopurinol is 1 - 3:1. More specifically, the mass ratio of cannabidiol to allopurinol can be 1:1, 1.5:1, 2:1, 2.5:1, 3:1, 3.5:1, 4:1, 4.5:1, etc., or any interval formed by the above mass ratios. The present invention is not limited thereto.

[0018] Furthermore, the present invention claims the use of the combination of cannabidiol and allopurinol in the preparation of a drug for reducing organ damage caused by hyperuricemia.

[0019] Furthermore, the present invention claims the use of the combination of cannabidiol and allopurinol in the preparation of a drug for treating hyperuricemia and reducing organ damage caused by hyperuricemia.

[0020] In some embodiments, the organ damage is kidney damage.

[0021] In some embodiments, the mass ratio of cannabidiol to allopurinol is 0.8 - 5:1.

[0022] In some embodiments, cannabidiol further includes its pharmaceutically acceptable salts or co-crystals. Pharmaceutically acceptable salts include conventional salts formed with pharmaceutically acceptable inorganic acids or organic acids, or inorganic bases or organic bases.

[0023] Compared with the prior art, the present invention has the following beneficial effects:

[0024] The present invention discovers through research that when cannabidiol and allopurinol are used in combination, they can synergistically improve the effect of reducing uric acid or treating hyperuricemia; at the same time, when cannabidiol and allopurinol are used in combination, they can synergistically reduce the levels of serum uric acid and serum creatinine, and thus play a synergistic role in reducing uric acid and / or treating hyperuricemia and synergistically reducing organ damage caused by hyperuricemia. The combination of cannabidiol and allopurinol used in the present invention can synergistically exert the effects of enhancing efficacy and reducing toxicity, providing a new idea for the design, synthesis and development of new uric acid-lowering drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 Effect of cannabidiol on the expression level of ABCG2 protein in HT-29 cells.

[0026] Figure 2 Effect of cannabidiol on the levels of serum uric acid, fecal uric acid and urinary uric acid in hyperuricemic mice. Among them, Figure 2 A shows the effect of cannabidiol on the serum uric acid level in hyperuricemic mice; Figure 2 B shows the effect of cannabidiol on the fecal uric acid level in hyperuricemic mice; Figure 2Effect of cannabidiol on urine uric acid level in hyperuricemia mice.

[0027] Figure 3 Effect of cannabidiol on protein expression levels of ABCG2 and URAT1 in intestine and kidney of hyperuricemia mice. Among them, Figure 3 A is the effect of cannabidiol on protein expression level of ABCG2 in intestine of hyperuricemia mice; Figure 3 B is the effect of cannabidiol on protein expression levels of ABCG2 and URAT1 in kidney of hyperuricemia mice.

[0028] Figure 4 H&E staining images of kidney tissue sections of mice in each group.

[0029] Figure 5 Effect of each group on joint inflammation in gout rat models. Among them, Figure 5 A is the ankle swelling condition of rats in each group; Figure 5 B is the ankle circumference difference of rats in each group 6 hours after modeling; Figure 5 C is the ankle circumference difference of rats in each group 12 hours after modeling.

[0030] Figure 6 Effect of each experimental group on uric acid level and kidney injury level in mice. Among them, Figure 6 A is the effect of each experimental group on serum uric acid in mice; Figure 6 B is the effect of each experimental group on urine uric acid in mice; Figure 6 C is the effect of each experimental group on fecal uric acid in mice; Figure 6 D is a schematic diagram of the appearance of the kidneys of the experimental group; Figure 6 E is the effect of each experimental group on serum creatinine in mice. Detailed implementation manners

[0031] The present invention will be further described below in conjunction with the accompanying drawings of the specification and specific embodiments, but the embodiments do not impose any form of limitation on the present invention. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the technical field.

[0032] Solution preparation:

[0033] Preparation of sodium carboxymethylcellulose (CMC-Na) solution: Weigh 2.5 g of sodium carboxymethylcellulose and dissolve it in 500 mL of sterile physiological saline to prepare a 5% sodium carboxymethylcellulose solution.

[0034] Preparation of potassium oxonate solution: Weigh potassium oxonate at a dose of 350 mg / kg and dissolve it in the above 5% sodium carboxymethylcellulose solution.

[0035] Preparation of cannabidiol (CBD) solution: Weigh cannabidiol at doses of 0.1 mg / kg and 0.5 mg / kg respectively, and dissolve it in the above-mentioned 5% sodium carboxymethyl cellulose solution containing dimethyl sulfoxide.

[0036] Preparation of allopurinol (AP) solution: Weigh allopurinol at a dose of 5 mg / kg and dissolve it in the above-mentioned 5% sodium carboxymethyl cellulose solution.

[0037] Example 1 Effect of cannabidiol on regulating ABCG2 in vitro

[0038] 1) Incubate HT-29 cells with high-concentration uric acid and intervene with cannabidiol

[0039] When the HT-29 cells in the culture dish grow to 90% density, passage them. Aspirate the old medium in the culture dish, add 500 μL of trypsin, let it stand for 30 s, aspirate the trypsin, add 1 mL of complete medium to collect the cells, dilute the cell suspension to a certain multiple, and add it into a 6-well plate for culture. When the cells in the 6-well plate grow to 70% density, replace the complete medium containing 0 - 15 μmol / L cannabidiol and incubate for 2 h, then add 14 mg / dL uric acid and continue to culture for 10 h.

[0040] 2) Detect the protein expression level of ABCG2 in HT-29 cells by western blot

[0041] Aspirate the complete medium in the cells and wash them once with PBS; place the cell culture dish or well plate on ice, add a certain amount of RIPA buffer, collect the cell lysate and perform protein quantification. Centrifuge the above cell lysate at 15,000 g and 4 °C for 10 min, then separate the supernatant for quantification. Dilute the protein supernatant 10 times with RIPA Buffer in a 96-well plate. Mix solution A and solution B of the BCA protein quantification kit in proportion, then mix it with the diluted protein supernatant, incubate at 37 °C for 30 min, use an enzyme-linked immunosorbent assay (ELISA) reader to detect the absorbance and calculate the RIPA Buffer and 5×LoadingBuffer required to quantify to the same protein concentration, and add them to the protein samples in turn. Heat the quantified protein samples at 100 °C for 5 min. The quantified protein samples are subjected to western blot detection or stored in a -80 °C refrigerator for a long time.

[0042] Figure 1 For the effect of cannabidiol on the protein expression level of ABCG2 in HT-29 cells. As Figure 1As shown, the relative protein expression level of ABCG2 in the cells of the uric acid group (UA group) incubated alone was significantly lower than that of the blank group (P<0.01); under the intervention of 5-15 μM cannabidiol, the relative protein expression level of ABCG2 was significantly increased compared with that of the UA group (P<0.001), suggesting that cannabidiol can promote the protein expression of ABCG2 in HT-29 cells in a concentration-dependent manner.

[0043] Example 2 In vivo uric acid-lowering activity test of cannabidiol

[0044] 1) Grouping of experimental animals

[0045] Twenty-four male Kunming mice weighing 18-22 g were purchased from the animal center of Southern Medical University. The mice were given standard feeding conditions: temperature 25±1°C, humidity 50±10%, 12-hour light / dark cycle, and free access to feed and water. After one week of adaptive feeding, the 24 mice were randomly divided into 4 groups: blank group (Control), hyperuricemia model group (HUA), CBD administration group (HUA+CBD), and positive control group (allopurinol administration group, HUA+AP). The number of mice in each group was 6.

[0046] 2) Establishment of hyperuricemia mouse model and administration intervention of cannabidiol and allopurinol

[0047] After one week of adaptive feeding of the mice, the mice in the HUA+CBD group and the HUA+AP group were pre-administered at 9:00 am every day for 21 consecutive days. Among them, the intragastric administration dose of the CBD administration group was 10 mg / kg, and the intragastric administration dose of the positive control group was 10 mg / kg. The mice in the Control group and the HUA group were intragastrically administered with normal saline containing 2% Tween 80 (0.1 mL / 10 g body weight). One hour after intragastric administration, the mice in the HUA group, the CBD administration group, and the allopurinol administration group were intraperitoneally injected with PO to induce hyperuricemia, and the PO injection dose was 350 mg / kg, while the mice in the Control group were intraperitoneally injected with 0.5% CMC-Na, and the dose was 0.1 mL / 10 g body weight.

[0048] 3) Collection of urine and feces samples and detection of their uric acid content

[0049] On the 20th day of the animal experiment, after intragastric administration and intraperitoneal injection, all mice were placed in a metabolic cage, and urine and feces were collected for 6 hours. After the urine samples were collected, they were centrifuged at room temperature (4000g, 10 min), the supernatant samples were separated and diluted 10-fold with ddH2O, and the uric acid concentration was detected using a uric acid kit. An equal amount of fecal samples from each mouse was weighed, an equal amount of ddH2O was added, vortexed, and sonicated using an ultrasonic instrument to make it fully dispersed. After centrifugation at 4000g at room temperature for 10 min, the supernatant was taken, and the uric acid concentration was detected using a uric acid kit.

[0050] 4) Animal euthanasia and detection of biochemical indicators

[0051] On the 21st day of the animal experiment, two hours after drug administration, the mice were anesthetized, blood was collected by removing the eyeballs, and the supernatant was taken after centrifugation of the blood samples. Relevant biochemical indicators were detected using a biochemical analyzer. Subsequently, the cervical vertebrae of the mice were severed for euthanasia, the kidneys and intestines were dissected and taken out, the kidney tissues were fixed with 4% paraformaldehyde solution, and the remaining organ tissues were stored in a -80°C refrigerator for subsequent detection.

[0052] Figure 2 Effects of cannabidiol on serum uric acid, fecal uric acid, and urine uric acid levels in hyperuricemic mice. As Figure 2 shown in A, on the 21st day of drug administration, the serum uric acid value of the model group was significantly higher than that of the blank group (P<0.001); compared with the model group, the serum uric acid value of the CBD (10 mg / kg) administration group was significantly decreased (P<0.001); the serum uric acid level of the allopurinol (10 mg / kg) administration group was also significantly decreased compared with the model group (P<0.001), and the mean value was lower than that of the CBD administration group. This result suggests that CBD (10 mg / kg) has obvious in vivo uric acid-lowering activity.

[0053] As Figure 2 shown in B, after the 20th day of drug administration, 6h feces of each group of mice were collected and the uric acid content was detected. The fecal uric acid content of the model group was slightly higher than that of the blank group (P<0.05); compared with the model group, the fecal uric acid content of the CBD (10 mg / kg) administration group was significantly increased (P<0.001), while there was no statistical difference in the fecal uric acid content of the allopurinol (10 mg / kg) administration group. It can be seen that the CBD administration group can reduce the serum uric acid level by promoting the excretion of uric acid in the intestine.

[0054] As Figure 2As shown in Figure C, 6-hour urine samples were collected from mice in each group 20 days after drug administration, and the uric acid content was measured. By measuring the urine volume and urine uric acid concentration of each mouse, the total amount of urinary uric acid excretion was calculated. The total amount of urinary uric acid excretion in the model group was significantly higher than that in the blank group (P<0.001); compared with the model group, the total amount of urinary uric acid excretion in the CBD (10 mg / kg) administration group was significantly increased; while there was no significant difference in the total amount of urinary uric acid excretion between the allopurinol (10 mg / kg) administration group and the model group.

[0055] These results indicate that in hyperuricemic mice, due to the increase in serum uric acid levels, the amount of uric acid excreted through urine also increases. The CBD administration group can further promote the excretion of uric acid through urine in the renal tubules, thereby reducing the serum uric acid level, but the allopurinol administration group cannot affect the amount of urinary uric acid excretion.

[0056] Example 3 Regulation of ABCG2 and URAT1 by Cannabidiol in Vivo

[0057] 1) Protein extraction and quantification from mouse intestinal and kidney tissues

[0058] Cut 10 mg of kidney tissue and place it in a 1.5 mL thick-walled centrifuge tube. Add 500 μL of RIPA Buffer and homogenize with a Servicebio animal tissue homogenizer for 2 min; cut a 2-cm ileum section above the cecum. After removing the intestinal contents, place the intestinal tube in a 1.5 mL thick-walled centrifuge tube, add 500 μL of RIPA Buffer, and homogenize with an animal tissue homogenizer for 2 min. Take the above homogenate for protein quantification, and the quantification steps are the same as those described in Test Example 1.

[0059] 2) Detection of the expression levels of ABCG2 and URAT1 in tissues by western blot

[0060] Use western blot to detect the protein expression level of ABCG2 in the protein sample of mouse intestinal tissue and the protein expression levels of ABCG2 and URAT1 in the protein sample of kidney tissue.

[0061] Figure 3 This is the effect of cannabidiol on the protein expression levels of ABCG2 and URAT1 in the intestine and kidney of hyperuricemic mice. As Figure 3 shown in Figure A, compared with the blank group, the protein expression of ABCG2 in the ileum of the model group mice was significantly inhibited (P<0.01), while the CBD (10 mg / kg) administration group could restore the protein expression of ABCG2 to a level comparable to that of the blank group.

[0062] As Figure 3As shown in Figure B, compared with the blank group, the expression of ABCG2 protein in the kidneys of the model group was significantly inhibited (P<0.001), while the relative protein expression level of ABCG2 in the kidneys of the CBD (10 mg / kg) administration group was significantly increased compared with the model group (P<0.001); the relative protein expression level of URAT1 in the kidneys of the model group was significantly increased compared with the blank group (P<0.001), while the CBD (10 mg / kg) administration group restored the protein expression of URAT1 to the level of the blank group.

[0063] Example 4 Protective Effect of Cannabidiol on the Kidneys

[0064] HE staining of kidney tissue sections: Take kidney samples from the same side and fix them by soaking in 4% paraformaldehyde. Dehydrate them step by step with ethanol solutions of different concentrations (70%-100%), and the dehydration time for each ethanol concentration is 40 min. After clearing with xylene, prepare paraffin sections. After the paraffin sections are stained with hematoxylin and eosin successively, dehydrate and mount them. Observe the sections with an electron microscope.

[0065] Figure 4 H&E staining diagrams of kidney tissue sections of mice in each group. As Figure 4 shown, the morphology of renal tubular epithelial cells in the blank group of mice was normal, while the model group showed obvious edema of renal tubular epithelial cells, renal tubular dilation, and inflammatory cell infiltration, and the CBD (10 mg / kg) administration group significantly improved the above pathological morphological changes; however, compared with the HUA group, the allopurinol (10 mg / kg) administration group showed a pathological condition similar to that of the model group and did not show an improvement effect.

[0066] Example 5 Effect of Cannabidiol on Improving Joint Inflammation in a Gout Rat Model

[0067] 1) Grouping of experimental animals

[0068] Select 32 SPF-grade male SD rats (200±20 g). Under the alternating conditions of a temperature of (23±2)°C, a relative humidity of 50%-70%, and a 12-h day-night cycle of indoor light, after adaptive feeding for 7 days, randomly divide them into 4 groups: blank group (n = 8), gout model group (n = 8), colchicine administration group (0.6 mg / kg, n = 8), and cannabidiol administration group (5 mg / kg, n = 8).

[0069] 2) Establishment of a gout model in the rat ankle joint and administration intervention with cannabidiol and colchicine

[0070] Before modeling, rats in each drug administration group were intragastrically administered the corresponding drugs at 10:00 am every day for three consecutive days. One night before modeling, the rats were weighed, grouped, and numbered, and the hair on the thigh was shaved to expose the ankle joint. At 8:00 am on the fourth day, each group of rats was intraperitoneally injected with 10% chloral hydrate (3 mL / kg) for anesthesia. After anesthesia, 50 μL of monosodium urate suspension was injected into the right ankle joint of the rats in the model group and each drug administration group for modeling, and 50 μL of PBS was injected into the right ankle joint of the rats in the blank group. One hour after modeling, each drug administration group was intragastrically administered the corresponding drugs, and the blank group and the model group were injected with the corresponding dose of solvent. After modeling, the toe swelling degree and gait of the rats were closely observed. At 0, 6, and 12 hours after modeling, the ankle joint circumference of the rats at different time periods was measured with a white line and a ruler, marked on the white line, and recorded in the table in real time. At the same time, the swelling of the ankle joints of the rats in each group was photographed, and the background, shooting distance, and ankle joint position were unified for shooting.

[0071] Figure 5 It is a schematic diagram of the effect of cannabidiol on improving joint inflammation in a gouty rat model. In the figure, * indicates P < 0.05 for comparison between two groups, ** indicates P < 0.01 for comparison between two groups, and *** indicates P < 0.001 for comparison between two groups.

[0072] As Figure 5 shown in

[0073] A, there were no obvious changes in the ankle joints of the rats in the blank group, while the right ankle joints of the rats in the gout model group showed obvious swelling at 6 and 12 hours after modeling. Compared with the gout model group, the degree of ankle joint swelling in the colchicine and cannabidiol administration groups was significantly smaller. Figure 5 As

[0074] shown in Figure 5 B, at 6 hours after modeling, the difference in the ankle joint circumference between the left and right ankles of the rats in the colchicine group was significantly lower than that in the gout model group; the difference in the ankle joint circumference between the left and right ankles of the rats in the cannabidiol group was also significantly lower than that in the gout model group, but the mean value of the circumference difference was higher than that in the colchicine group.

[0075] Example 6 Cannabidiol combined with allopurinol plays a synergistic and toxicity-reducing role in the treatment of hyperuricemia

[0076] 1) Experimental animal grouping

[0077] Twenty-four SPF male KM mice (20±2 g) were selected. Under the alternating conditions of temperature (23±2)°C, relative humidity 50% - 70%, and indoor light duration of 12 h day-night cycle, after 7 days of adaptive feeding, they were randomly divided into 4 groups: uric acid nephropathy model group (n = 6), cannabidiol administration group (5 mg / kg, n = 6), allopurinol administration group (5 mg / kg, n = 6), allopurinol combined with cannabidiol administration group (cannabidiol 5 mg / kg, allopurinol 5 mg / kg, n = 6).

[0078] 2) Establishment of hyperuricemic nephropathy mouse model and drug intervention

[0079] Each administration group was given the corresponding drug by gavage at 0.1 ml / 10 g at 9:30 am every day. The uric acid nephropathy model group was given the same volume of 0.5% sodium carboxymethylcellulose by gavage. All mice were intraperitoneally injected with potassium oxonate (400 mg / kg) at 0.1 mL / 10 g at 10:00 am and given hypoxanthine (500 mg / kg) by gavage. The experiment lasted for 7 days. After the administration ended on the 7th day, the mice were sacrificed by blood collection. After dissection, the kidney tissues were collected and stored at -80°C.

[0080] The drug preparations and concentrations of each group were as follows: 0.5% sodium carboxymethylcellulose was used as the drug solvent, potassium oxonate (40 mg / mL), hypoxanthine (50 mg / mL), cannabidiol (0.5 mg / mL), allopurinol (0.5 mg / mL). The injection was given at a volume of 100 μL / 10 g.

[0081] 3) Sample treatment and determination

[0082] After the model was established on the sixth day, metabolic cages were used to collect feces and urine for 6 h, and the uric acid concentrations in feces and urine were detected respectively. On the seventh day, blood was collected by eye enucleation 1 h after the model was established. After the blood was allowed to stand for 30 min, it was centrifuged at 3000 g for 10 min at room temperature, and the supernatant was taken to measure the serum uric acid and creatinine concentrations. Subsequently, the mice were sacrificed by cervical dislocation, the kidneys were dissected and the appearance of each mouse's kidney was photographed and recorded.

[0083] Figure 6 For the effects of cannabidiol combined with allopurinol on the uric acid level and kidney injury level in mice. In the figure, * indicates P < 0.05 for comparison between two groups, ** indicates P < 0.01 for comparison between two groups, and *** indicates P < 0.001 for comparison between two groups.

[0084] As Figure 6As shown in Figure A, either 5 mg / kg cannabidiol alone or 5 mg / kg allopurinol can significantly reduce the serum uric acid level in hyperuricemic nephropathy mice. The effect of reducing the serum uric acid level in hyperuricemic nephropathy mice by combining 5 mg / kg allopurinol with cannabidiol is superior to that of using 5 mg / kg cannabidiol alone or 5 mg / kg allopurinol respectively. It can be seen that the combination of allopurinol and cannabidiol can synergistically reduce the serum uric acid in hyperuricemic nephropathy mice.

[0085] As Figure 6 shown in Figure B and Figure 6 Figure C, using cannabidiol alone can significantly increase the uric acid levels in urine and feces.

[0086] As Figure 6 shown in Figure D, the kidneys of mice in the model group and the allopurinol administration group showed varying degrees of paleness and swelling in appearance, while the kidney damage in the cannabidiol administration group was improved.

[0087] As Figure 6 shown in Figure E, the serum creatinine levels in the group using cannabidiol alone and the group using allopurinol alone were significantly decreased compared with the model group, and the ability of the allopurinol combined with cannabidiol group to reduce the serum creatinine level was superior to that of the group using cannabidiol alone or the group using allopurinol alone respectively. This indicates that the combination of allopurinol and cannabidiol can synergistically reduce kidney damage. The above results show that cannabidiol can reduce the serum uric acid level by increasing the excretion of uric acid from urine and feces, and can show the effect of enhancing efficacy and reducing toxicity when combined with allopurinol.

[0088] The foregoing embodiments are merely illustrative and are used to explain some features of the method of the present invention. The appended claims are intended to claim the broadest possible scope that can be conceived, and the embodiments presented herein are demonstrated by the applicant's actual test results. Therefore, the applicant's intention is that the appended claims are not limited by the selection of examples that illustrate the features of the present invention. Some numerical ranges used in the claims also include sub-ranges within them, and variations within these ranges should also be interpreted as being covered by the appended claims whenever possible.

Claims

1. Use of the combination of cannabidiol and allopurinol in the preparation of a drug for reducing uric acid and / or treating hyperuricemia.

2. The use according to claim 1, characterized in that, The reduction of uric acid refers to reducing the uric acid in the serum.

3. The use according to claim 2, wherein Reducing the uric acid in the serum means promoting the expression of ABCG2 and / or down-regulating the expression of URAT1 to thereby reduce uric acid.

4. The use according to claim 1, wherein The hyperuricemia includes gout or gout complications caused by hyperuricemia.

5. The use according to claim 4, wherein, The gout complications are one or more of gouty arthritis and urinary calculi.

6. According to the use described in claim 1, it is characterized in that, The hyperuricemia is hyperuricemia caused by potassium oxonate and hypoxanthine.

7. According to the use described in claim 1, it is characterized in that, The mass ratio of cannabidiol to allopurinol is 0.8 - 5:

1.

8. Use of the combination of cannabidiol and allopurinol in the preparation of a drug for reducing organ damage caused by hyperuricemia.

9. The use according to claim 8, characterized in that, The organ damage is kidney damage.

10. According to the use described in claim 8, it is characterized in that, The mass ratio of cannabidiol to allopurinol is 0.8 - 5:1.

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