A small intestinal mucosal extract with preventive and therapeutic effects on bipolar disorder

Abstract

The present invention belongs to the field of food and medicine, and specifically relates to a small intestinal mucosa extract having a preventive and therapeutic effect on bipolar disorder. The present invention also provides a health product or medicine containing the small intestinal mucosa extract. Through pharmacological experiments, it can be found that compared with existing treatment methods, the small intestinal mucosa extract provided by the present invention not only has a significant bidirectional regulatory effect on the depressive episodes and manic episodes of bipolar disorder, but also has the advantages of no toxic side effects and preventing repeated attacks. The small intestinal mucosa extract method provided by the present invention is simple and easy to implement, and is suitable for further industrialized large-scale production.

Description

Technical Field

[0001] The invention belongs to the field of medicine, and in particular relates to a composition having preventive and therapeutic effects on bipolar disorder. Background Art

[0002] Bipolar disorder is one of the common mental disorders in clinical practice. It is characterized by complex condition, long course, difficult treatment, easy recurrence, and high disability rate. It is usually accompanied by symptoms of alternating depressive episodes and manic episodes, which in turn has a great adverse effect on the patient's physical and mental health. With the continuous increase in people's life and work pressure in recent years, the rate of depressive episodes in bipolar disorder has shown an increasing trend year by year. In order to promptly improve the patient's depressive symptoms and stabilize the patient's emotional and psychological state, it is necessary to provide them with drug treatment with few adverse reactions, high safety and good efficacy, so as to effectively improve the patient's physical and mental state and improve their survival and quality of life.

[0003] In the medical field, psychological nursing intervention combined with drug care is usually used to help patients reduce or even eliminate their symptoms of both mania and depression. From the perspective of pharmacology, there is great dialectic in the use of drugs for patients with mental illness. Sedatives are often used for manic patients, and antidepressants are used for depression. The antagonism of the two drugs cannot be ignored. In addition, when diagnosing the disease, there is a misdiagnosis of bipolar disorder as unipolar depression. Therefore, for patients with bipolar disorder, starting from the diagnosis of the disease, their treatment is not an easy task. Therefore, finding an effective drug that has a significant therapeutic effect on bipolar disorder and can reduce its recurrence rate has become the primary task of pharmacists.

[0004] The small intestine is the most important part of the food digestion and absorption process. Jame (1997) pointed out that mucosal digestion is the final digestion stage of various nutrients, and the intestinal mucosa is the final digestion site of all nutrients, which plays a key role. In addition to its digestion and absorption functions, the small intestine is also an important endocrine organ. A variety of active substances have been isolated from the small intestine and its mucosa, including heparin sodium, various enzymes and antibacterial substances. The various biological activities of small intestinal mucosal extracts will surely become a hot topic for pharmacists in the future. Summary of the invention

[0005] One of the purposes of the present invention is to provide a new use of a small intestinal mucosa extract, namely, the small intestinal mucosa extract has a preventive and / or therapeutic effect on bipolar disorder; and has a significant improvement effect on the depression, mania, learning and memory ability, and insomnia of patients with bipolar disorder.

[0006] The small intestinal mucosa extract of the invention is prepared from the small intestinal mucosa through extraction and precipitation processes.

[0007] The extraction method may be one or more of water extraction and ultrasonic extraction; preferably, the extraction method is ultrasonic extraction.

[0008] The above-mentioned precipitation method can be one or more of ethanol precipitation method, graded precipitation method, and quaternary ammonium salt complexation method; preferably, the precipitation method is ethanol precipitation method.

[0009] Specifically, the preparation method of the small intestinal mucosa extract comprises the following steps:

[0010] (1) Extraction: Take the small intestinal mucosa, homogenize and crush it, add water and then perform ultrasonic extraction, filter the extract to obtain the water extract;

[0011] (2) alcohol precipitation: adding monohydric alcohol to the aqueous extract obtained in step (1), and filtering to obtain a precipitate;

[0012] (3) Freeze-drying: The precipitate obtained in step (2) is freeze-dried to obtain a small intestinal mucosal extract containing mucopolysaccharides and water-soluble proteins.

[0013] Wherein, the small intestinal mucosa in step (1) is selected from the small intestinal mucosa of mammals; preferably, the small intestinal mucosa is porcine small intestinal mucosa.

[0014] Preferably, the volume mass ratio of water to small intestinal mucosa used in ultrasonic water extraction in step (1) is 1:1-100:1 (ml / g);

[0015] Further preferably, the volume mass ratio of water used to small intestinal mucosa is 80:1 (ml / g).

[0016] Preferably, after the monohydric alcohol is added in step (2), the alcohol concentration of the solution is 25%-95%;

[0017] More preferably, the added monohydric alcohol is ethanol, and the ethanol concentration of the solution is 65%.

[0018] Therapeutic effects of small intestinal mucosal extract on bipolar disorder.

[0019] The results of Experimental Example 1 show that the small intestinal mucosal extract provided by the present invention can significantly improve the depressive state of patients with bipolar disorder and can significantly improve the learning and memory abilities of patients with bipolar disorder.

[0020] The results of Experimental Example 2 show that the small intestinal mucosal extract provided by the present invention can significantly improve the manic state of patients with bipolar disorder and can significantly improve the insomnia state of patients with bipolar disorder.

[0021] Clinical trial results show that the intestinal mucosal extract provided by the present invention not only has significant efficacy and few toxic side effects in treating patients with bipolar disorder, but also has the advantage of preventing relapse compared with other therapeutic drugs.

[0022] The present invention also provides a medicine comprising the composition. Preferably, the medicine can be a tablet, a granule, a capsule or a syrup, and can be prepared according to a common preparation process.

[0023] Preferably, the human dosage of the above-mentioned drug for the treatment of bipolar disorder is 0.02 mg / kg-200 mg / kg, and more preferably, the human dosage is 0.5 mg / kg-50 mg / kg.

[0024] The medicine of the present invention may include only active ingredients or may include excipients, antioxidants, pH regulators, preservatives, isotonic agents and any other pharmaceutically acceptable auxiliary materials with guaranteed safety.

[0025] The medicine of the present invention can be prepared into various types, including not only tablets, granules, capsules and syrups, but any other changes to the external form of the product are technical solutions that can be associated with those skilled in the art, so changing the shape of the final product falls within the scope of the present invention.

[0026] Compared with the prior art, the small intestinal mucosal extract provided by the present invention has the following main advantages in preventing and treating bipolar disorder:

[0027] (1) It has significant therapeutic effects, no toxic side effects, and can be taken for a long time.

[0028] (2) Compared with other treatment methods, the medicine containing the small intestinal mucosa extract of the present invention has the advantage of effectively preventing the recurrence of bipolar disorder. Specific embodiments

[0029] In order to enable those skilled in the art to fully understand the present invention, the present invention is further described below through specific examples, but those skilled in the art should be aware that the examples of the present invention do not limit the present invention in any way.

[0030] Example 1 Preparation of small intestinal mucosal extract

[0031] (1) Water extraction: Take the pig small intestinal mucosa, homogenize it, add water at a volume-to-mass ratio of 50:1 (ml / g), extract it in a 100°C water bath, and filter the extract to obtain a water extract;

[0032] (2) alcohol precipitation: adding ethanol to the aqueous extract obtained in step (1) so that the ethanol concentration in the solution is 45%, and filtering to obtain a precipitate;

[0033] (3) Freeze-drying: The precipitate obtained in step (3) is freeze-dried to obtain a small intestinal mucosal extract containing mucopolysaccharides and water-soluble proteins.

[0034] Example 2 Preparation of small intestinal mucosal extract

[0035] (1) Extraction: Take the pig small intestinal mucosa, homogenize and crush it, add water at a volume-to-mass ratio of 10:1 (ml / g), perform ultrasonic extraction, and filter the extract to obtain a water extract;

[0036] (2) alcohol precipitation: adding ethanol to the aqueous extract obtained in step (1) so that the ethanol concentration in the solution is 95%, and filtering to obtain a precipitate;

[0037] (3) Freeze-drying: The precipitate obtained in step (3) is freeze-dried to obtain a small intestinal mucosal extract containing mucopolysaccharides and water-soluble proteins.

[0038] Example 3 Preparation of small intestinal mucosal extract

[0039] (1) Extraction: Take the pig small intestinal mucosa, homogenize and crush it, add water at a volume-to-mass ratio of 100:1 (ml / g), perform ultrasonic extraction, and filter the extract to obtain a water extract;

[0040] (2) Alcohol precipitation: adding ethanol to the aqueous extract obtained in step (1) so that the ethanol concentration in the solution is 25%, and filtering to obtain a precipitate;

[0041] (3) Freeze-drying: The precipitate obtained in step (3) is freeze-dried to obtain a small intestinal mucosal extract containing mucopolysaccharides and water-soluble proteins.

[0042] Example 4 Preparation of small intestinal mucosal extract

[0043] (1) Extraction: Take the pig small intestinal mucosa, homogenize and crush it, add water at a volume-to-mass ratio of 80:1 (ml / g), perform ultrasonic extraction, and filter the extract to obtain a water extract;

[0044] (2) Alcohol precipitation: adding ethanol to the aqueous extract obtained in step (1) so that the ethanol concentration in the solution is 65%, and filtering to obtain a precipitate;

[0045] (3) Freeze-drying: The precipitate obtained in step (3) is freeze-dried to obtain a small intestinal mucosal extract containing mucopolysaccharides and water-soluble proteins.

[0046] Example 5 Preparation of small intestinal mucosal extract

[0047] (1) Extraction: Take the pig small intestinal mucosa, homogenize and crush it, add water at a volume-to-mass ratio of 60:1 (ml / g), perform ultrasonic extraction, and filter the extract to obtain a water extract;

[0048] (2) alcohol precipitation: adding ethanol to the aqueous extract obtained in step (1) so that the ethanol concentration in the solution is 45%, and filtering to obtain a precipitate;

[0049] (3) Freeze-drying: The precipitate obtained in step (3) is freeze-dried to obtain a small intestinal mucosal extract containing mucopolysaccharides and water-soluble proteins.

[0050] Example 6 Preparation of a drug containing a small intestinal mucosal extract

[0051] Take the small intestinal mucosal extract and starch obtained in Example 4, sieve and mix; add starch slurry to make a soft material, sieve, granulate, dry and granulate; then add magnesium stearate, mix and tablet, and obtain tablets.

[0052] Experimental Example 1 Effect of the Composition on Improving Depressive Behavior in Depression Model Rats

[0053] 1. Test methods

[0054] 80 healthy male SD rats were randomly divided into a normal control group, a model group and an experimental group (Example 3 group, Example 4 group), 20 rats in each group. The model group and the experimental group rats were raised in solitary and combined with chronic mild unpredictable stimulation according to the reference method to construct a depression rat model, and the blank control group was raised normally. During the modeling period, the experimental group was gavaged with the small intestinal mucosal extract obtained in Example 3 and Example 4 at 20 mg / kg, and the control group and the model group were gavaged with an equal amount of 0.9% NaCl for 4 consecutive weeks. The model group rats were accompanied by typical depression symptoms such as reduced water intake, exploratory behavior, weight loss, decreased interest, and dark fur color.

[0055] 2. Detection indicators

[0056] (1) Field experiment:

[0057] After the administration, the open field experiment was carried out according to the reference method. The number of horizontal movements and vertical upright times of each group of rats within 4 minutes were observed and recorded. The sum of the horizontal and vertical scores was calculated as the number of spontaneous activities, and the immobility time was also recorded.

[0058] (2) Morris water maze test to detect learning and memory function of rats

[0059] After the administration, the rats in each group were subjected to positioning navigation experiment and space exploration experiment using Morris water maze test system according to the reference (ApoE4 age-dependently increases the susceptibility to stress-induced depressive-like behavior and the decline of learning and memory ability).

[0060] 3. Test results

[0061] (1) Effects of small intestinal mucosal extract on depressive behavior in rats with depression

[0062] The number of spontaneous activities of rats in the model group was less than that of the normal control group (P < 0.01), while that of the experimental group was more than that of the model group (P < 0.01); the immobility time of rats in the model group was longer than that of the normal control group (P < 0.01), while that of the experimental group was shorter than that of the model group (P < 0.01), indicating that the small intestinal mucosal extract can significantly improve the depressive state of rats with depression model. The specific results are shown in Table 1.

[0063] Table 1 Effects of small intestinal mucosal extract on depressive behavior in rats with depression (n=20)

[0064]

[0065] Compared with the normal control group, ## P<0.01; compared with the model group, && P<0.01.

[0066] (2) Effects of small intestinal mucosal extract on learning and memory abilities of rats with depression

[0067] The latency of the model group rats in the Morris water maze navigation test on the 1st, 2nd, 3rd, and 4th days was longer than that of the normal control group (P<0.05); while that of the experimental group was shorter than that of the model group (P<0.05); the number of crossings in the Morris water maze space exploration test in the model group was less than that in the control group (P<0.01), while that of the experimental group was more than that in the model group (P<0.05), indicating that the small intestinal mucosal extract can significantly improve the learning and memory ability of depression model rats. The specific results are shown in Table 2.

[0068] Table 2 Effects of small intestinal mucosal extract on learning and memory abilities of rats with depression (n=20)

[0069]

[0070] Compared with the normal control group, # P<0.05, ## P<0.01; compared with the model group, & P<0.05.

[0071] Experimental Example 2: Effect of the Composition on the Improvement of Depressive Behavior in Manic Model Rats

[0072] 1. Modeling method

[0073] 80 healthy SD rats were divided into a normal control group, a model group, and an experimental group (Example 3 group, Example 4 group) according to the random number table method, with 20 rats in each group. The model group and the experimental group used the modified multi-platform sleep deprivation method (MMPM) to establish a manic model (refer to the method in the reference document Changes in the expression of neuronal nitric oxide synthase after sleep deprivation and the effects of Xingnaojing on it). Rats can eat, drink and move on their own on the platform, and the water in the mouse box is changed every day. One week before the experiment, the rats were placed on the platform to adapt for 1 hour every day. After the experiment started, the rats were on the platform except for changing the water in the water tank, which lasted for 7 days. The normal control group used a mouse box of the same size as the model group, but the platform was not placed at the bottom, but a fine wire mesh was placed, the rats were placed on the net, water was placed under the net, and the water was 1 cm below the mesh, so that the rats could get normal sleep, and other conditions were the same.

[0074] 2. Intervention methods

[0075] The blank control group and the model group were not given any intervention treatment, while the experimental group was given 20 mg / kg of the small intestinal mucosal extracts obtained in Example 3 and Example 4 by oral gavage for intervention treatment. The intervention period was 4 weeks.

[0076] 3. Detection indicators

[0077] (1) Rat autonomous behavior measurement:

[0078] The Open-Field method (OIT) was used. One rat was placed in the central square of the open box each time. Each measurement lasted for 3 minutes. The horizontal score was the number of squares that the rat crossed horizontally. It was stipulated that 1 point was given when 3 or 4 paws of the rat entered the same square. The vertical score was given when the two hind paws supported the body and the two front paws left the bottom surface. Each time the paws left the bottom surface, 1 point was given.

[0079] (2) Sleep latency measurement

[0080] The rats were deprived of sleep for 7 consecutive days, and the time from the time the rat stepped off the platform to the time the rat fell asleep for the first time (i.e., sleep latency) was recorded after the model was completed. If it was greater than 30 minutes, the mania model was successfully established. The behavioral definition of rats falling asleep was that the rats closed their eyes and lay still for 30 seconds or more.

[0081] (3) Statistical methods

[0082] SPSS 20.0 statistical software was used for analysis and processing. Measurement data were expressed as mean ± standard deviation. Multiple groups were compared using one-way analysis of variance (ANOVA), and pairwise comparisons were performed using q test. P < 0.05 was considered statistically significant.

[0083] 4. Results

[0084] (1) Comparison of general behavior of rats in each group

[0085] Model group: After 1 day of sleep deprivation, the rats showed excitement, running around between different platforms, being sensitive to slight external sound and light stimulation, and showing panic to sound and light stimulation; after 3 days of sleep deprivation, they showed dullness, slight twitching of head and facial muscles accompanied by beard movement, squatting on the platform with their heads down, thinking about sleep, often falling into the water repeatedly due to napping, climbing back onto the platform after waking up, reducing activity and eating, and showing aggressive behaviors such as biting after stimulation. After 5-7 days of sleep deprivation, the rats were listless, had less eating activity, had erect hair all over their bodies, were extremely irritable, had increased aggressive biting behaviors, and 4 rats died during the biting process.

[0086] Blank control group: no manic manifestations of the model group.

[0087] Experimental group: Manic symptoms were significantly alleviated compared with the model group, and no animals died due to biting behavior.

[0088] The above results indicate that small intestinal mucosal extract can significantly improve the irritable and aggressive manic state of manic rats.

[0089] (2) Comparison of autonomous behavior results of rats in each group

[0090] Compared with the normal control group, the horizontal activity scores and vertical activity scores of the rats in the model group were significantly reduced (P<0.05), indicating that modeling can lead to a decrease in horizontal and vertical activities of rats. Compared with the model group, the horizontal activity scores and vertical activity scores of the small intestinal mucosal extract group were significantly higher than those of the model group (P<0.05), indicating that the small intestinal mucosal extract significantly improved the manic state of manic rats. The specific results are shown in Table 3.

[0091] Table 3 Effects of small intestinal mucosal extract on manic behavior in manic rats

[0092]

[0093] Compared with the normal control group, # P<0.05; compared with the model group, & P<0.05.

[0094] (3) Comparison of sleep latency in rats of each group after the experiment

[0095] Compared with the normal control group, the sleep latency of rats in the model group was significantly prolonged, with a statistically significant difference (P<0.05), indicating that modeling can lead to prolonged sleep latency in rats; compared with the model group, the sleep latency of the small intestinal mucosa extract group was significantly shortened, with a statistically significant difference (P<0.05), indicating that the small intestinal mucosa extract can significantly improve the insomnia state of manic rats. The specific results are shown in Table 4.

[0096] Table 4 Effects of small intestinal mucosal extract on sleep latency in manic rats

[0097]

[0098] Compared with the normal control group, # P<0.05; compared with the model group, & P<0.05.

[0099] Clinical Examples

[0100] 1.1 General Information

[0101] A total of 40 patients with bipolar disorder who were treated and treated from January to December 2016 were selected for the study and were divided into two groups according to the time of admission, namely the control group and the observation group. The 40 selected patients all met the clinical symptoms and diagnostic criteria for bipolar disorder manic episodes in CCMD-3. Patients with other systemic serious diseases, pregnant or lactating patients, and patients allergic to therapeutic drugs were excluded. The general information of the patients is shown in Table 5.

[0102] Table 5 General information of patients

[0103] Grouping Number of cases male female Average age Control group 20 11 9 40.12±6.09 Observation Group 20 12 8 39.88±4.45

[0104] Note: Comparison of patients’ general information, P>0.05.

[0105] 1.2 Methods

[0106] ①Control group: combined medication, oral quetiapine fumarate tablets, the first dose is 100 mg / day, gradually increased to 400-600 mg / day as the treatment progresses, and the maximum dose does not exceed 800 mg / day; oral sodium valproate tablets, the first dose is 200-400 mg / day, gradually increased to 600-1200 mg / day as the treatment progresses.

[0107] ②Observation group: The small intestinal mucosa extract of Example 3 was taken at 2 mg / kg, once a day.

[0108] The treatment course for both groups was 8 weeks.

[0109] 1.3 Observation indicators

[0110] The BRMS scoring scale was used to evaluate the mental symptoms of the two groups of patients before and after treatment, and the efficacy was determined. Significantly effective: the BRMS score before and after treatment decreased by 75%, and the clinical symptoms disappeared or improved significantly; effective: the BRMS score before and after treatment decreased by 50% to 75%, and the clinical symptoms improved; ineffective: the BRMS score before and after treatment decreased by less than 50%, and the clinical symptoms basically did not improve. Total effective rate = 1-ineffective rate. Adverse reactions (dizziness, drowsiness, nausea and vomiting, allergic reactions) of the two groups of patients were counted.

[0111] 1.4 Statistics of relapse rate after drug withdrawal

[0112] 1.5 Statistical methods

[0113] All data were analyzed using SPSS 20.0. Count data were expressed as (%), and P < 0.05 was considered statistically significant.

[0114] 2 Results

[0115] 2.1 Comparison of efficacy between the two groups of patients

[0116] The results showed that the overall effective rate of the observation group was significantly higher than that of the control group, and the difference between the two groups was significant and statistically significant. The specific results are shown in Table 6.

[0117] Table 6 Patient efficacy and comparison (n, %)

[0118] Grouping Number of cases Significant effect efficient invalid Total Effectiveness Control group 20 7(35％) 7(35％) 6(30％) 14(70％) Observation Group 20 11(55％) 7(35％) 2(10％) <![CDATA[18(90%) ﹩ ]]>

[0119] Note: Compared with the control group, ﹩ P<0.05.

[0120] 2.2 Comparison of adverse reactions in patients

[0121] The incidence of adverse reactions in the control group was 15% (1 case of dizziness, 1 case of drowsiness, 1 case of nausea and vomiting, a total of 3 cases); no adverse reactions such as dizziness, drowsiness, nausea and vomiting occurred in the observation group, and only 1 case of allergic reaction occurred, with an incidence of 5% (inter-group comparison, P < 0.05). This allergic reaction may be caused by impurities in the small intestinal mucosal extract, which can be avoided by increasing its purity.

[0122] 2.3 Comparison of relapse rate after drug discontinuation

[0123] After the 8-week treatment course, the patients continued to take the medicine for half a year to stabilize the condition. One year after stopping the medicine, 8 patients in the control group relapsed, with a relapse rate of 57.14%, while there was no relapse in the observation group.

[0124] The above results show that the composition not only has significant efficacy and few side effects in treating bipolar disorder, but also has the advantage of preventing relapse compared with other therapeutic drugs.

Claims

1. Use of a small intestinal mucosal extract in the preparation of a product for preventing and / or treating bipolar disorder, characterized in that: The small intestinal mucosa extract is a mixture containing mucopolysaccharides and water-soluble proteins. The small intestinal mucosa extract is prepared from pig small intestinal mucosa by ultrasonic water extraction and ethanol precipitation. The specific steps are: (1) extraction: take pig small intestinal mucosa, homogenize and crush it, add water and then perform ultrasonic extraction, the volume mass ratio of water to small intestinal mucosa is 1:1-100:1 in ml / g, and filter the extract to obtain a water extract; (2) alcohol precipitation: add ethanol to the water extract obtained in step (1) to make the ethanol concentration of the solution 25%-95%, and filter to obtain a precipitate; (3) freeze drying: freeze drying the precipitate obtained in step (2).

2. The use according to claim 1, characterized in that The use is use of the small intestinal mucosa extract in preparing a product for improving the depressive state in bipolar disorder.

3. The use according to claim 1, characterized in that The use is use of the small intestinal mucosa extract in preparing a product for improving the manic state in bipolar disorder.

4. The use according to claim 1, characterized in that The use is the use of the small intestinal mucosa extract in preparing a product for improving the learning and memory abilities in bipolar disorder.

5. The use according to claim 1, characterized in that The use is use of the small intestinal mucosa extract in preparing a product for improving insomnia in bipolar disorder.

6. The use according to any one of claims 1 to 5, characterized in that The dosage of the small intestinal mucosa extract is 0.02 mg / kg-200 mg / kg.

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