Application of ziziphus jujuba var. sarcodactylis in the preparation of drugs for the prevention and treatment of schizophrenia and for alleviating the side effects of olanzapine glucose and lipid metabolism.

Abstract

This invention discloses the application of catalpol in the preparation of drugs for the prevention and treatment of schizophrenia and the alleviation of the glucose and lipid metabolism side effects of olanzapine. Using a schizophrenia model mouse model induced by the commonly used MK801, the therapeutic effect of catalpol on schizophrenia was investigated, and the therapeutic effect of catalpol on the glucose and lipid metabolism side effects of the commonly used drug olanzapine was investigated through long-term gavage administration. A systematic study of catalpol was conducted from pharmacodynamic, behavioral, and morphological perspectives. The results showed that catalpol effectively improved the behavioral characteristics of the schizophrenia model mice, alleviated negative symptoms and cognitive impairment; catalpol effectively improved olanzapine-induced obesity and lipid metabolism disorders. This provides a theoretical research basis for the development of catalpol into a drug for the treatment of schizophrenia.

Description

Technical Field

[0001] This invention belongs to the pharmaceutical field, specifically relating to the application of catalpol in the preparation of drugs for the prevention and treatment of schizophrenia, and also relating to the application of catalpol in the preparation of drugs for alleviating the glucose and lipid metabolism side effects of olanzapine, a drug for schizophrenia. Background Technology

[0002] Schizophrenia is a chronic mental illness with a high relapse and disability rate, interfering with multiple cognitive functions, including memory, thinking, and perception. The lifetime prevalence of schizophrenia in China is 1.25%. Clinically, it manifests in three main categories: positive symptoms such as delusions, hallucinations, and thought disorders; negative symptoms such as fatigue, self-abandonment, social withdrawal, and poverty of thought; and cognitive impairment. Approximately 2.5% of the Chinese adult population suffers from lifelong mental disorders requiring lifelong treatment. Compared to healthy individuals, schizophrenia patients have a significantly higher incidence of comorbid physical illnesses and a higher mortality rate, and their life expectancy is shortened by 10–20 years.

[0003] Schizophrenia typically presents with positive symptoms, negative symptoms, and cognitive impairment. Positive symptoms are primarily behavioral and thought abnormalities, including delusions, hallucinations, and thought disorder; these symptoms often decrease over time. Negative symptoms are mainly emotional deprivation, including emotional flatness, poverty of speech, social impairment, and anhedonia. Cognitive impairment mainly includes difficulties with learning, memory, and problem-solving. Negative symptoms and cognitive impairment often remain stable or increase over time. Improving positive symptoms does not necessarily stabilize the condition; negative symptoms often exacerbate relapses. Compared to positive symptoms, negative symptoms more significantly impact a patient's quality of life, leading to social isolation, increased functional disorders, increased financial burden on families, and a worse prognosis. Negative symptoms are also the most burdensome symptoms for patients. Currently, clinical medications effectively improve the positive symptoms of schizophrenia, but the more far-reaching negative symptoms are often overlooked. Therefore, finding a drug treatment for the negative symptoms of schizophrenia is urgently needed.

[0004] Neurotransmitter abnormalities are a key aspect of the pharmacology of schizophrenia; therefore, clinical treatment of schizophrenia primarily relies on medication for remission. In 2001, the World Psychiatric Association (WPA) classified these drugs into first-generation antipsychotics (FGAs) and second-generation antipsychotics (SGAs) based on their pharmacological and clinical characteristics. First-generation antipsychotics are also known as classic antipsychotics, while second-generation antipsychotics are known as atypical antipsychotics. Second-generation (atypical) antipsychotics, such as olanzapine, are commonly used in clinical practice. Olanzapine is effective against positive symptoms of schizophrenia and has a low risk of extrapyramidal symptoms; therefore, it has gradually become a first-line drug for the treatment of schizophrenia. Although olanzapine has significantly improved the treatment outcomes and prognosis of patients with schizophrenia, long-term use of olanzapine can lead to increased weight and food intake, affecting tolerability and patient compliance, thus limiting its clinical application. Therefore, finding drugs to counteract the increased food intake associated with olanzapine is a clinical need.

[0005] Traditional Chinese medicine (TCM) has complex components and multiple targets, often playing a comprehensive role in treating diseases. Therefore, we are exploring substances from TCM that may be useful for treating schizophrenia. Catalpol belongs to the iridoid glycoside class of compounds and is one of the main components of Rehmannia glutinosa, a plant in the Scrophulariaceae family. Its molecular formula is C2. 15 H 22 O 10 Modern pharmacology has discovered that catalpol possesses various biological effects, including antioxidant activity, improved cognition, improved motor function, improvement of Alzheimer's disease, and antidepressant effects, which are consistent with some symptoms of schizophrenia. Currently, catalpol has not been used to treat schizophrenia or to mitigate its side effects. Summary of the Invention

[0006] In view of this, one objective of the present invention is to provide the use of catalpol in the preparation of medicaments for treating schizophrenia; another objective of the present invention is to provide the use of catalpol in the preparation of medicaments for alleviating the glucose and lipid metabolism side effects of olanzapine, a drug for treating schizophrenia.

[0007] To achieve the above objectives, the present invention provides the following technical solution:

[0008] 1. Application of catalpol in the preparation of antipsychotic drugs.

[0009] Preferably, the symptoms of schizophrenia in this invention are negative symptoms and cognitive impairment.

[0010] The preferred application of this invention is that of catalpol in the preparation of a drug that improves sensory and auditory gating deficits in patients with schizophrenia.

[0011] The preferred application of this invention is that of catalpol in the preparation of a drug that improves the cognitive abilities of patients with schizophrenia.

[0012] The preferred application of this invention is that of catalpol in the preparation of a drug for restoring the social function of patients with schizophrenia.

[0013] 2. Application of ziziphus tinctoria in the preparation of drugs that alleviate the glucose and lipid metabolism side effects of olanzapine, a drug for schizophrenia.

[0014] The preferred application of this invention is that of catalpol in the preparation of a drug to alleviate weight gain caused by olanzapine.

[0015] The preferred application of this invention is that of catalpol in the preparation of a drug to alleviate the hyperplasia and hypertrophy of abdominal fat cells caused by olanzapine.

[0016] The present invention preferably relates to the use of catalpol in the preparation of drugs that inhibit the accumulation of lipid droplets in hepatocytes, thereby alleviating olanzapine-induced fatty vacuolation and fatty degeneration of liver tissue.

[0017] The beneficial effects of this invention are as follows:

[0018] This invention discloses the application of catalpol in the preparation of antipsychotic drugs and in improving the side effects of olanzapine. Catalpol has a significant effect on the negative symptoms of schizophrenia in mouse models; it also has a significant proliferative effect on isolated liver cells, significantly improves olanzapine-induced glucose and lipid metabolism disorders in rats, and reverses liver lipid metabolism in rat models of glucose and lipid metabolism disorders. Therefore, catalpol can be used to prepare antipsychotic drugs, particularly for treating negative symptoms of schizophrenia and preventing and treating olanzapine-induced glucose and lipid metabolism disorders. This invention provides a new clinical use for catalpol, expanding its application scope. Attached Figure Description

[0019] To make the objectives, technical solutions, and beneficial effects of this invention clearer, the following figures are provided for illustration:

[0020] Figure 1 Example 1: Comparison of PPI test results of low, medium and high PPI groups in schizophrenic mice with the model group (Saline is physiological saline, MK801 is diazepam, Cata is catalpol, OLZ is olanzapine, *p<0.05, **p<0.01);

[0021] Figure 2 Example 2: Comparison of the results of the novel object recognition experiment in the low, medium and high group treated with catalpol with the model group;

[0022] Figure 3 Example 3: Comparison of the temporal memory test results of the low, medium and high levels of catalpol in schizophrenic mice with the model group;

[0023] Figure 4 Example 4: Comparison of the results of the three-box social test in schizophrenic mice treated with catalpol in low, medium and high groups with the model group;

[0024] Figure 5 Example 5: Comparison of the results of the rotarod test in normal mice treated with haloperidol in the low, medium and high groups with the model group (Hal is haloperidol, *p<0.05, **p<0.01);

[0025] Figure 6 Example 6 shows the effect of catalpol on olanzapine-induced weight gain in rats (OLZ represents olanzapine, CAT represents catalpol, and Sim represents simvastatin; compared with the model group, *p<0.05, **p<0.01; compared with the olanzapine group, #p<0.05, ##p<0.01).

[0026] Figure 7 The results of the detection of the effects of olanzapine and catalpol on the food / water intake of rats in Example 6 were compared with those of the model group (OLZ = olanzapine, CAT = catalpol, Sim = simvastatin, *p<0.05, **p<0.01; compared with the olanzapine group, #p<0.05, ##p<0.01).

[0027] Figure 8 Example 6: Effect of olanzapine on adipocyte diameter and the ameliorative effect of catalpol (CAT is catalpol, Sim is simvastatin; compared with the model group, *p<0.05, **p<0.01; compared with the olanzapine group, #p<0.05, ##p<0.01).

[0028] Figure 9 To investigate the effects of olanzapine on liver tissue morphology and the ameliorative effect of catalpol in Case 6 (CAT represents catalpol, Sim represents simvastatin; compared with the model group, *p<0.05, **p<0.01; compared with the olanzapine group, #p<0.05, ##p<0.01);

[0029] Figure 10 To investigate the effects of olanzapine on hepatocyte steatosis and the ameliorative effect of catalpol (CAT for catalpol, Sim for simvastatin; compared with the model group, *p<0.05, **p<0.01; compared with the olanzapine group, #p<0.05, ##p<0.01). Detailed Implementation

[0030] The present invention will be further described below in conjunction with the accompanying drawings and specific embodiments, so that those skilled in the art can better understand the present invention and be able to implement it. However, the exemplified embodiments are not intended to limit the present invention.

[0031] The catalpol used in the present invention was purchased from Nanjing Jingzhu Biotechnology Co., Ltd., CAS No.: 2415-24-9, batch number: JZ21082501.

[0032] The animals used in the present invention were purchased from Chongqing Lepeter Biotechnology Co., Ltd., production license number: SCXK(Xiang)2019-0004, and the experimental animal quality certificate number was No.430727220101944023. They were housed in the Experimental Animal Center of the School of Pharmacy, Southwest University (Chongqing, China). After one week of adaptive feeding, they entered the experiment. During the animal experiment, the experimental animals were treated well and strictly raised in accordance with the relevant regulations on experimental animal science and management.

[0033] The positive drug olanzapine used in the present invention is one of the commonly used clinical drugs for treating schizophrenia. Its English name is Olanzapine, and its chemical name is 2-methyl-4-(4-methyl-1-piperazinyl)-10H-thieno[2,3-b][1,5]benzodiazepine.

[0034] Example 1. Prepulse Inhibition Experiment

[0035] Catalpol treats prepulse inhibition deficits in schizophrenic mice. Three days in advance, the mice were placed in the experimental chamber to adapt to the 70 dB background noise for 5 minutes. On the day of the formal experiment, the mice were placed in the laboratory half an hour in advance to adapt to the environment. The start and end of the experiment were four 120 dB startle stimuli (40 ms), and these startle stimuli were not used for data analysis. After the four startle stimuli, the animals randomly received 16 startle stimuli, 10 no-stimulus trials, and 30 PPI trials. Each PPI trial had prepulses of 73, 76, or 82 dB (i.e., 3, 6, 12 dB higher than the background), lasting 20 ms. Then there was a 40 ms startle stimulus (120 dB), with a fixed interval of 80 ms. The three PPI trials (73, 76, and 82 dB) were each conducted 10 times, with an inter-trial interval of 10-20 seconds, and the experiment lasted for 23 minutes.

[0036] The prepulse inhibition (PPI) experiment in rodents is often used to simulate the sensory auditory gating deficits in schizophrenia patients and is often used to study antipsychotic drugs. Prepulse Inhibition (PPI) refers to presenting a weaker stimulus (prepulse) within 30-500 ms before the occurrence of a stronger stimulus (pulse), thereby inhibiting the startle response to the stronger stimulus.

[0037] The specific grouping, dosage, and administration method of the catalpol treatment for the low, medium, and high levels of prepulse inhibition in schizophrenic mice are shown in Table 1.

[0038] Table 1

[0039]

[0040] Experimental results are as follows Figure 1 As shown, under sound stimulation of 73, 76, and 82 dB, different doses of catalpol could improve the pre-pulse inhibition defect in schizophrenic mice, with the best therapeutic effect observed at a dose of 5 mg / kg. This indicates that catalpol can improve sensory and auditory gating defects in schizophrenia.

[0041] Example 2. New Object Recognition Experiment

[0042] For the first three days of the experiment, mice were allowed to freely explore the center of an open space for 5 minutes each day. On the day of the experiment, mice were placed in an open area 5 cm away from a wall with two objects of the same size, color, and shape. After the drug took effect, the mice were placed in the center of the two objects and allowed to explore freely for 5 minutes. After 30 minutes, one of the objects was replaced with a new one, and the mice were placed in the center of the two objects and allowed to explore freely for another 5 minutes. The exploration time of the mice for the new and old objects was recorded using a double-blind method, and the discrimination index was calculated. Discrimination index (DI) = (exploration time of new object - exploration time of old object) / (exploration time of new object + exploration time of old object) x 100%.

[0043] The specific grouping, dosage, and administration method of ziziphus alcohol in the low, medium, and high groups for the novel object recognition experiment in schizophrenic mice are shown in Table 2.

[0044] Table 2

[0045]

[0046] Experimental results are as follows Figure 2 As shown, the time required for exposure to new objects was significantly longer in the control group compared to the model group, indicating that the MK801 model of cognitive deficit was successfully established. The time required for exposure to new objects was significantly longer in the catalpol treatment group compared to the model group. This indicates that catalpol can improve the cognitive abilities of schizophrenic mice, with the best improvement observed at 5 mg / kg.

[0047] Example 3. Temporal Memory Experiment

[0048] Three days before the experiment, mice were placed in the center of an open field for 10 minutes. On the day of the experiment, mice were placed in the laboratory beforehand to acclimatize. Two identical objects (Object 1) were then placed in the open field, with the mice positioned in the center of the open field and allowed to explore freely for 10 minutes. After a 30-minute interval, the objects were replaced by a new object (Object 2), and the mice were placed in the center of the open field from the same position and direction and explored freely for 10 minutes. After another 30-minute interval, Object 2 was replaced with Object 1 and placed in the same position, and the mice were placed in the center of the open field from the same position and direction and explored freely for 5 minutes. The time the mice spent interacting with the two objects was recorded, and the discrimination index (DI) was calculated. The discrimination index (DI) was calculated as: (Object 1 exploration time - Object 2 exploration time) / (Object 1 exploration time + Object 2 exploration time) x 100%.

[0049] For the specific grouping, dosage, and administration method of the catalpol treatment for the low, medium, and high levels of schizophrenic mice in the temporal memory experiment, please refer to Table 2.

[0050] Experimental results are as follows Figure 3 As shown, compared with the model group, the time spent in contact with object 1 in the catalpol treatment group was significantly longer. Compared with the normal group, there was no significant difference in the time spent in contact with object 1 in the catalpol treatment group. This indicates that catalpol can improve the cognitive abilities of schizophrenic mice.

[0051] Example 4. Three-Box Social Experiment

[0052] An empty cage E was placed in each of the left and right side chambers, 5 cm away from the walls of the experimental chamber. The experimental mouse was allowed to explore freely in the middle chamber for 10 minutes. Then, a strange mouse S1 of the same sex and similar age and weight was introduced into the empty cage. The experimental mouse was allowed to explore freely in the middle chamber for 10 minutes. The close contact time between the experimental mouse and the strange mouse S1 and the empty cage E on the left was recorded in the second stage. Close contact was defined as a distance of less than or equal to 3.5 cm. Then, a new strange mouse S2 of the same sex and similar age and weight was introduced into another empty cage. The experimental mouse was allowed to explore freely in the middle chamber for 10 minutes. The close contact time between the experimental mouse and the strange mice S1 and S2 in the third stage was recorded, and the mouse's discrimination index was calculated. Discrimination index (DI) = (exploration time of new object - exploration time of old object) / (exploration time of new object + exploration time of old object) x 100%.

[0053] For the specific grouping, dosage, and administration method of the three-box social experiment in schizophrenic mice treated with ziziphus acetylcholine in low, medium, and high groups, please refer to Table 2.

[0054] Experimental results are as follows Figure 4As shown, in the second stage, the model group showed no significant preference for the unfamiliar mouse S1 compared to the empty cage E, indicating that MK801 can impair the social function of mice, and MK801 successfully created a social impairment model; citronella did not cause social dysfunction in mice. In the second stage, compared to the model group, the citronella treatment group showed a significantly increased contact time with the unfamiliar mouse S1 and an increased preference index, suggesting that the schizophrenic mice treated with citronella exhibited normal sociality. In the third stage, we found that the control group, model group, and citronella treatment group showed no preference for the unfamiliar mouse S2, indicating that the social novelty of the schizophrenic mice was not restored. Overall, the results show that citronella treatment restored the sociality of schizophrenic mice but not their social novelty.

[0055] Example 5. Rotating bar experiment

[0056] Two days before the start of the experiment, the mice underwent acclimatization training. They were trained twice daily on a rotating apparatus at 10 rpm for 5 minutes each time, with a 30-minute interval between training sessions. During the formal experiment, the mice were placed on an acceleration lever that increased from 0 to 20 rpm within 20 seconds, and the experiment lasted for 5 minutes. The time of the first fall and the number of falls within 5 minutes were recorded using a double-blind method.

[0057] The specific grouping, dosage, and administration method of the low, medium, and high concentrations of catalpol in the rotarod experiment on normal mice are shown in Table 3.

[0058] Table 3

[0059]

[0060] Experimental results are as follows Figure 5 As shown, compared with the positive control group, the drop time in the catalpol group was significantly longer, while the drop frequency was significantly lower. Compared with the normal group, there was no significant difference in drop time or drop frequency in the catalpol group. This indicates that catalpol does not induce extrapyramidal symptoms in mice, meaning it does not have the extrapyramidal symptoms associated with first-generation antipsychotic drugs.

[0061] Example 6

[0062] Table 4 shows the grouping, dosage, and administration method of ziziphus acetonide for the prevention of lipid metabolism disorders induced by olanzapine.

[0063] Table 4

[0064]

[0065]

[0066] During the administration of the drug to rats, their body weight, food intake, and water consumption were recorded every other day. After the administration was completed, samples were collected for subsequent experiments.

[0067] like Figure 6 As shown, at the beginning of the experiment, there was no significant difference in the average body weight of the six groups of rats (P > 0.05). One week after administration, the weight gain of the rats in each group began to change. The weight gain rate of rats given only olanzapine gradually increased compared to the blank control group. The weight gain of rats in the groups given different doses of catalpol combined with olanzapine was also higher than that of the blank control group, but lower than that of the model group. Among them, there was no significant difference in weight gain between the 30 mg / kg and 60 mg / kg catalpol groups, and the weight gain was slightly lower than that of the 120 mg / kg catalpol group.

[0068] By recording the food and water intake of each group of rats at each time, it was found that, Figure 7 As shown, compared with the blank control group, the average food intake and water consumption of rats in the model group increased significantly every two days (P < 0.05). However, there was no significant difference in food intake between the treatment groups and the model group (P > 0.05), while water consumption was significantly reduced in all treatment groups (P < 0.05). These results indicate that intervention with different doses of citronella did not significantly reduce the amount of food consumed by rats, but rather reduced water consumption. This suggests that olanzapine causes side effects such as obesity or weight gain in rats, and these adverse reactions are associated with food intake and water consumption. It also suggests that citronella can effectively alleviate these adverse reactions, but perhaps by reducing water consumption rather than controlling the amount of food consumed.

[0069] HE staining is commonly used to observe the morphology of tissue sections. The HE staining results and statistical findings of white abdominal fat sections from each group of rats are shown below. Figure 8 As shown in the figure. Compared with the blank control group, the model group showed a significant increase in the diameter of abdominal adipocytes in female SD rats after administration of olanzapine at a dose of 4 mg / kg (P < 0.05). Statistical results showed that under the presence of olanzapine, intervention with simvastatin and catalpol at doses of 30 mg / kg, 60 mg / kg, and 120 mg / kg respectively resulted in a decreasing trend in adipocyte diameter, with a significant difference between the simvastatin and 120 mg / kg catalpol groups (P < 0.05). The results indicate that olanzapine can induce obesity and cause adipocyte hyperplasia and hypertrophy, while catalpol significantly improved the adipocyte enlargement induced by olanzapine, suggesting that catalpol may have a certain therapeutic effect on lipid metabolism imbalance.

[0070] HE staining was performed on liver tissue to observe whether the morphology of hepatocytes in each group of rats was intact and whether the cell nuclei were enlarged. At the same time, the damaging or protective effects of drugs and other stimulating factors on the liver were detected. Figure 9This image shows HE staining of the right lobe of the rat liver in each group, observed under a 400× microscope. No significant enlargement of liver cells was observed in any group. However, compared to the control group, the olanzapine group exhibited significant pathological changes in the liver. Hepatocyte nuclei were surrounded by balloon-like transparent fat vacuoles of varying sizes, distributed throughout the cytoplasm, which merged to form large vacuoles, pushing the nuclei towards the cell membrane. Statistical results showed that intervention with the positive control drug simvastatin and different doses of catalpol significantly reduced fat vacuolation. Furthermore, the area of ​​the vacuoles gradually decreased with increasing catalpol dose. This indicates that olanzapine can induce severe fat vacuolation in liver tissue and may lead to hepatocyte steatosis. High doses of catalpol and simvastatin have significant preventive and therapeutic effects on olanzapine-induced hepatic steatosis.

[0071] Oil Red O staining is commonly used to detect fat levels in tissues or cells. The results of Oil Red O staining of the right lobe of the liver in each group of rats are shown below. Figure 10 As shown in the figure. Statistical results showed that, compared with the control group, in the model group, after administration of olanzapine alone, lipid droplets were distributed throughout the entire cytoplasm of the liver, with a statistically significant difference (P < 0.01). Furthermore, compared with the model group, low, medium, and high doses of citronellal and simvastatin combined with olanzapine significantly reduced the distribution and content of intracellular lipids, with statistical significance (P < 0.01). This indicates that olanzapine can significantly promote the accumulation of lipid droplets in hepatocytes, and citronellal has a similar effect to the positive control drug simvastatin, showing a significant improvement after drug intervention.

[0072] In summary, this invention investigated the therapeutic effect of catalpol on schizophrenia using a mouse model of schizophrenia induced by the commonly used MK801 method, and further examined its therapeutic effect on the glucose and lipid metabolism side effects of the commonly used drug olanzapine through long-term gavage administration. A systematic study of catalpol was conducted from pharmacodynamic, behavioral, and morphological perspectives. The results showed that catalpol effectively improved the behavioral characteristics of the schizophrenia model mice, alleviated negative symptoms and cognitive impairment; and effectively improved olanzapine-induced obesity and lipid metabolism disorders. This provides a theoretical research basis for developing catalpol into a drug for the treatment of schizophrenia.

[0073] The above-described embodiments are merely preferred embodiments provided to fully illustrate the present invention, and the scope of protection of the present invention is not limited thereto. Equivalent substitutions or modifications made by those skilled in the art based on the present invention are all within the scope of protection of the present invention. The scope of protection of the present invention is defined by the claims.

Claims

1. The application of catalpol in the preparation of drugs for treating schizophrenia, characterized in that: The symptoms of schizophrenia are negative symptoms and cognitive impairment.

2. The application according to claim 1, characterized in that: The application of ziziphus tinctoria in the preparation of drugs that improve sensory and auditory gating deficits in patients with schizophrenia.

3. The application according to claim 1, characterized in that: The application of ziziphus tinctoria in the preparation of drugs that improve cognitive abilities in patients with schizophrenia.

4. The application according to claim 1, characterized in that: Application of ziziphus jujuba var. spinosa in the preparation of drugs to restore the social function of patients with schizophrenia.

5. Application of ziziphus jujuba var. spinosa in the preparation of drugs that alleviate the glucose and lipid metabolism side effects of olanzapine, a drug for schizophrenia.

6. The application according to claim 5, characterized in that: The use of ziziphus tinctoria in the preparation of drugs to alleviate weight gain caused by olanzapine.

7. The application according to claim 5, characterized in that: Application of ziziphus tinctoria in the preparation of drugs to alleviate olanzapine-induced abdominal fat cell hyperplasia and hypertrophy.

8. The application according to claim 5, characterized in that: The application of ziziphus jujuba var. spinosa in the preparation of drugs that inhibit the accumulation of lipid droplets in hepatocytes, thereby alleviating olanzapine-induced fatty vacuolation and fatty degeneration in liver tissue.

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