CN114732826A - Application of gamma-aminobutyric acid and spinosyn in prevention, alleviation or treatment of anxiety - Google Patents
Application of gamma-aminobutyric acid and spinosyn in prevention, alleviation or treatment of anxiety Download PDFInfo
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- A61K31/00—Medicinal preparations containing organic active ingredients
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- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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- A61K31/195—Carboxylic acids, e.g. valproic acid having an amino group
- A61K31/197—Carboxylic acids, e.g. valproic acid having an amino group the amino and the carboxyl groups being attached to the same acyclic carbon chain, e.g. gamma-aminobutyric acid [GABA], beta-alanine, epsilon-aminocaproic acid or pantothenic acid
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Abstract
The invention discloses application of gamma-aminobutyric acid and spinosin in prevention, alleviation or treatment of anxiety disorder. The invention prepares the product with the functions of preventing, relieving or treating the anxiety by compounding the gamma-aminobutyric acid and the spinosad, can achieve better effect under less active ingredient dosage, reduces the dosage of medicines or health-care products, particularly can reduce the dosage of the spinosad with higher cost, reduces the production cost of the medicines or the health-care products, and is expected to become a new anti-anxiety medicine or health-care product with high efficiency and low cost.
Description
Technical Field
The invention relates to application of gamma-aminobutyric acid (GABA) and Spinosin in synergy in preventing, relieving or treating anxiety, and belongs to the technical field of medicine preparation and mental disease prevention and treatment.
Background
Anxiety disorder (anxiety), also called anxiety neurosis, is the most common one of the major diseases of neurosis, is characterized by anxiety mood experience, comprises anxiety disorder caused by a plurality of causes such as generalized anxiety disorder, panic attack, social anxiety disorder and the like, is a heterogeneous disease, and therefore, a new medicine needs to be developed aiming at the anxiety disorder with different causes. Among them, generalized anxiety disorder is the most common anxiety disorder, and Generalized Anxiety Disorder (GAD) is a chronic mental disease that presents extensive and excessive anxiety to events in life, and is the most common anxiety disorder, mainly represented by: there is no clear and objective stress worry, restlessness, and vegetative nerve dysfunction symptoms such as palpitation, trembling hand, sweating, frequent micturition, and restlessness. The latest Chinese study on mental disorder disease burden and health service utilization shows that the prevalence rate of anxiety disorder is the highest among various mental disorders, the lifetime prevalence rate is 7.6%, and the 12-month prevalence rate is 5.0%; wherein the GAD lifetime prevalence rate is 0.3%, and 12-month prevalence rate is 0.2%; the number of newly increased patients in China exceeds 240 million every year. With the acceleration of life rhythm and the increase of social pressure, anxiety people are continuously expanded. In the clinic, psychological and pharmacological treatments are commonly used for anxiety disorders. Currently, the first-line clinical anxiolytics mainly include benzodiazepine drugs, Selective 5-hydroxytryptamine Reuptake Inhibitor (SSRI), 5-hydroxytryptamine And norepinephrine Reuptake Inhibitor (SNRI), And the like. The benzodiazepine drug has quick response, but short effect duration, and can also cause addiction after long-term use. SSRI and SNRI have slow effect (effect is 2-3 weeks), and need long-term use, and sudden or unreasonable withdrawal of the medicine is easy to appear and withdraw the medicine, and some patients with mild anxiety have a concern for using the medicine. Moreover, these drugs may not be effective in some patients and are clinically referred to as drug resistant anxiety disorders. Therefore, it is necessary to search for a more effective drug with less side effects for treating anxiety.
Gamma-aminobutyric acid (GABA) is an important central nervous system inhibitory neurotransmitter, has good water solubility and thermal stability, and has a structural formula shown in the specification. GABA, a small molecular weight nonprotein amino acid, has been confirmed to be safe for eating and useful for the production of foods such as functional beverages. The research shows that the ingestion of a certain amount of GABA has the physiological effects of improving anxiety, sleeping and the like. The gamma-aminobutyric acid can be prepared by a microbial fermentation method, can be produced at a lower cost, and has the advantages of high yield and good safety.
Spinosin (Spinosin) is one of the main extracts of spina date seeds, and has the following structural formula. The spina date seed is a common traditional tranquilizer in China, and is widely used for treating various syndromes and diseases such as insomnia, neurasthenia, climacteric syndrome and the like. Studies suggest that the spinosad has a strong effect of improving anxiety.
Although anxiolytic effects of both have been reported, whether a synergistic effect of the combination of both is obtained has not been investigated. Gamma-aminobutyric acid and spinosin are used as food-grade components, no reports on liver and kidney function damage or addiction exist at present, and if the anxiolytic effects of the Gamma-aminobutyric acid and the spinosin are improved, a safer and more effective anxiolytic option can be provided for the anxiety people. In addition, the production cost of the spinosad is higher than that of GABA, and if a formula combination with synergistic interaction can be found, the production cost can be controlled on the premise of ensuring the product efficacy, so that the spinosad is more beneficial to industrial popularization and application.
Disclosure of Invention
The invention discovers through experimental research that the combined use of gamma-aminobutyric acid (GABA) and Spinosin is more effective than the single use of GABA or Spinosin in the overhead cross test of a mammal and a mouse, and the combined use of the GABA and the Spinosin has synergistic effect on the prevention, the relief and the treatment of the anxiety disorder.
Based on the research results, the invention provides an application of gamma-aminobutyric acid and spinosad in coordination in preparing a product for preventing, relieving or treating anxiety.
In the application, the mass ratio of the gamma-aminobutyric acid to the spinosin is 1: 0.2 to 5, preferably 1: 0.5-2, more preferably 1: 1. For example, the mass ratio of γ -aminobutyric acid to spinosin may be 1: 0.2, 1: 0.3, 1: 0.4, 1: 0.5, 1: 0.6, 1: 0.7, 1: 0.8, 1: 0.9, 1: 1.0, 1: 1.2, 1: 1.4, 1: 1.6, 1: 1.8, 1: 2.0, 1: 2.2, 1: 2.4, 1: 2.6, 1: 2.8, 1: 3.0, 1: 3.2, 1: 3.4, 1: 3.6, 1: 3.8, 1: 4.0, 1: 4.2, 1: 4.4, 1: 4.6, 1: 4.8, 1: 5.0 or any range therebetween.
In the above use, the content of gamma-aminobutyric acid and spinosin in the product for preventing, alleviating or treating anxiety disorder is therapeutically effective amount. The therapeutically effective amount can be adjusted by one skilled in the art according to the subject to be administered, for example, the total effective amount per single use of gamma-aminobutyric acid and spinosyn can be 10mg/kg, 20mg/kg, 30mg/kg, 40mg/kg, 50mg/kg, 60mg/kg, 70mg/kg, 80mg/kg, 90mg/kg, 100mg/kg, 150mg/kg, 200mg/kg, etc.
In the above applications, the content of gamma-aminobutyric acid and spinosin in the product for preventing, alleviating or treating anxiety disorder in unit dose is therapeutically effective amount.
In the application, the product for preventing, relieving or treating the anxiety neurosis can be prepared into various feasible formulations, including but not limited to injections, solutions, granules, powders, tablets, capsules, pills, oral liquids, suspensions, sprays, freeze-dried powder injections and the like.
In the application, the product for preventing, relieving or treating the anxiety neurosis is a medicine or a health-care product, and has the effects of preventing, relieving, improving, treating and the like on the anxiety neurosis.
In the application, the product for preventing, relieving or treating the anxiety can be applied to the human body and can also be applied to the animal body. When used in animals, the animal may be mouse, rabbit, etc.
In one embodiment of the present invention, when the product for preventing, alleviating or treating anxiety disorders is used in mice, the total effective single use amount of gaba and spinosyn is 40 mg/kg.
The invention also provides a product for preventing, relieving or treating anxiety, and the active ingredients of the product comprise gamma-aminobutyric acid and spinosin.
In the product for preventing, relieving or treating anxiety, the mass ratio of the gamma-aminobutyric acid to the spinosin is 1: 0.2 to 5, preferably 1: 0.5-2, more preferably 1: 1.
The product for preventing, relieving or treating anxiety can be a medicine or a health product.
In the product for preventing, relieving or treating anxiety neurosis, the content of gamma-aminobutyric acid and the spinosin in the product is effective amount for treatment.
In the above products for preventing, alleviating or treating anxiety disorders, the amount of gamma-aminobutyric acid and spinosin in the unit dosage product is a therapeutically effective amount.
In the product for preventing, relieving or treating anxiety disorder, the active ingredients can also contain other ingredients with the effects of preventing, treating, relieving or improving anxiety disorder or other ingredients with other effects besides gamma-aminobutyric acid and spinosin.
The product for preventing, relieving or treating anxiety disorder also comprises pharmaceutically acceptable adjuvants, which are adjuvants required for preparing various dosage forms, such as dispersant, binder, disintegrant, lubricant, bulking agent, solvent, cosolvent, surfactant, sweetener, etc.
In the product for preventing, relieving or treating anxiety neurosis, the dosage form can be various feasible dosage forms, including but not limited to injection, solution, granules, powder, tablets, capsules, pills, oral liquid, suspension, spray and freeze-dried powder injection.
Furthermore, the product for preventing, relieving or treating anxiety disorder has the effects of preventing, relieving, improving and treating the anxiety disorder. The product can be used for human body, and also can be used for animal body. When used in animals, the animal may be mouse, rabbit, etc. When the product is used for mice, the total effective usage amount of the gamma-aminobutyric acid and the spinosin in a single time is 40 mg/kg.
The invention discovers that the combined use of gamma-aminobutyric acid (GABA) and Spinosin has a synergistic effect on anxiety, and provides a new idea for the research and development of products for preventing, relieving and treating the anxiety for the first time. The product prepared by compounding the gamma-aminobutyric acid (GABA) and the Spinosin has better effect of preventing, relieving or treating the anxiety disorder compared with the single product of the gamma-aminobutyric acid (GABA) or the Spinosin.
The invention compounds gamma-aminobutyric acid (GABA) and Spinosin to prepare the medicine or health care product, can achieve better effect with less active ingredient dosage, reduces the dosage of the medicine or health care product, particularly can reduce the dosage of the Spinosin with higher cost, reduces the production cost of the medicine or health care product, and is expected to become a new anxiolytic medicine or health care product with high efficiency and low cost.
Drawings
FIG. 1 shows the experimental results of the mouse elevated plus maze;
in fig. 1, ordinate: elevated plus maze experiment open arm time(s), abscissa: grouping; from left to right, sequentially forming a blank comparison group and a CRS molding group; CRS +20mg/kg GABA +20mg/kg spinosin formula group; CRS +40mg/kg of spinosin group; CRS +40mg/kg GABA group. (Note: CRS means a chronic restrictive restraint molding method). *:p＜0.05；**：p＜0.01；****：p＜0.0001。
Detailed Description
The invention is described in detail below with reference to the drawings and the detailed description, it being understood that the following examples are illustrative and explanatory only and are not restrictive of the scope of the invention in any way. In the following embodiments, the biochemical reagents not specifically described are all conventional reagents in the art, and may be prepared according to conventional methods in the art or commercially available, and may be of laboratory pure grade.
Example 1 investigation of the synergistic Effect of GABA and Spirosin combination on anxiolytic Effect
Experimental reagent:
GABA CAS No.: 56-12-2, molecular formula: C4H9NO2, available from Huaxi Biotech, Inc.; spinosin, CAS No.: 72063-39-9, formula: C28H32O15, available from Phyllomycete, Inc.
Experimental animals:
c57BL/6 mice, male, weighing 20-26 grams, provided by wi-tongli-hua biotechnology limited, license number: SCXK (Kyoto) 2021-. The animals are raised in cages with the light and shade period of 12h/12h, the room temperature of 20-22 ℃, the water is free, and the feed is provided by the Huada protein experimental animal center.
Experimental grouping:
after the experimental animals are adapted for one week, the animals are divided into 5 groups, each group comprises 8-15 animals, and each group comprises: blank control (no molding); a CRS group; CRS +20mg/kg GABA +20mg/kg spinosin group; CRS +40mg/kgGABA group; CRS +40mg/kg spinosin group. (Note: CRS means a chronic restrictive binding modeling method)
Molding method and grouping treatment:
an anxiety model is manufactured by adopting a chronic restrictive constraint modeling method (CRS), and the method comprises the following steps: all CRS molded mice were placed in a well-ventilated 50 ml centrifuge tube (with vent holes) for 2 hours a day, and were continuously restrained for 15 days, and the mice were allowed to move in the limited space in the tube without causing visible damage, and after the restraint was completed, the mice were returned to normal diet in a mouse cage. The placebo mice were housed in normal cages under normal conditions.
While molding for 15 days, each group of mice was treated as follows: mice were gavaged daily with saline 0.1ml/10g for the placebo group and the sham group (CRS group). The CRS +20mg/kg GABA +20mg/kg spinosin group was administered with a physiological saline containing GABA and spinosin dissolved in a 1:1 mass ratio, and the mice were gavaged with a solution of 0.1ml/10g body weight, the gavage amount of GABA was 20mg/kg body weight, and the gavage amount of spinosin was 20mg/kg body weight. CRS +40 mg/kgGABA: GABA is dissolved in normal saline to prepare 4g/L solution, and the mouse is subjected to intragastric administration according to the weight of 0.1ml/10 g. CRS +40mg/kg Spinosad group: the spinosyns were dissolved in physiological saline to make a 4g/L solution, and the mice were gavaged with 0.1ml/10g body weight.
Mouse elevated plus maze experiment:
the next day after 15 days CRS modeling, the mouse elevated plus maze experiment was performed. On the next day after the molding is finished, intragastric administration is carried out on each group of mice according to a grouping treatment mode, and after 60 minutes of intragastric administration, the mice are subjected to an elevated plus maze experiment; at the start of the experiment, mice were placed in the maze from the central compartment towards the open arm, and the activity of each mouse was recorded over 6 minutes using the Any-maze software, with the residence time of the mice in the open arm being observed primarily. After the experiment is completed, the mouse is taken out, the two arms are cleaned, alcohol is sprayed to remove smell, and then the next mouse experiment is continued. The stay time into the open arm is inversely related to the anxiety mood of the mouse, and the shorter the stay time, the more severe the anxiety mood of the mouse.
The statistical method comprises the following steps: the results are expressed as mean ± SE, with multiple samples tested by ANOVA and two samples tested by t.
The experimental results are as follows: as shown in fig. 1, the open arm residence time in the CRS group was significantly lower than that of the blank control group, indicating that the CRS group mice were significantly anxious after CRS modeling. The open arm retention time of the 3 mice treated with GABA40mg/kg, spinosin 40mg/kg and GABA20mg/kg + spinosin 20mg/kg was higher than that of the CRS group (p< 0.05), which means that the 3 interventions can significantly relieve the anxiety mood of mice after CRS modeling, wherein the GABA20mg/kg + spinosin 20mg/kg has the most significant effect (p< 0.0001). In 3 treatment groups of GABA40mg/kg, spinosyn 40mg/kg and GABA20mg/kg + spinosyn 20mg/kg, the anxiety capability of GABA20mg/kg + spinosyn 20mg/kg for alleviating CRS modeling of mice is respectively and remarkably higher than that of GABA40mg/kgp＜0.01）And spinosin 40mg/kg (p< 0.05), which indicates that the combination of GABA20mg/kg and spinosin 20mg/kg has a synergistic effect on anxiolytic effect, i.e. the combination of GABA and spinosin 1:1 has an unexpected synergistic effect compared with GABA and spinosin alone.
Example 2 screening of compounding ratio of GABA and spinosin
Experimental reagent:
GABA CAS No. γ -aminobutyric acid: 56-12-2, molecular formula: C4H9NO2, available from Huaxi Biotech, Inc.; spinosin, CAS No.: 72063-39-9, formula: C28H32O15, available from Phyllomycete, Inc.
Experimental animals:
c57BL/6 mice, male, weighing 20-26 grams, provided by wi-tongli-hua biotechnology limited, license number: SCXK (Jing) 2021-. The animals are raised in cages with the light and shade period of 12h/12h, the room temperature of 20-22 ℃, the water is free, and the feed is provided by the Huada protein experimental animal center.
Grouping experiments:
after the experimental animals are adapted for one week, the animals are divided into 11 groups, each group comprises 8-15 animals, and each group comprises: blank control (no molding); a CRS group; CRS + GABA: spinosin 1:1 group; CRS + GABA: spinosin 1:2 group; CRS + GABA: spinosin 1:5 group; CRS + GABA: spinosin 1: 8; CRS + GABA: spinosin 2:1 group; CRS + GABA: spinosin 5:1 group; CRS + GABA: spinosin 8:1 group; CRS +40mg/kgGABA group; CRS +40mg/kg spinosin group. (Note: CRS means a chronic restrictive binding modeling method)
Molding method and grouping treatment:
an anxiety model is manufactured by adopting a chronic restrictive constraint modeling method (CRS), and the method comprises the following steps: all CRS molded mice were placed in a well-ventilated 50 ml centrifuge tube (with vent holes) for 2 hours a day, and were continuously restrained for 15 days, and the mice were allowed to move in the limited space in the tube without causing visible damage, and after the restraint was completed, the mice were returned to normal diet in a mouse cage. The placebo mice were housed in normal cages under normal conditions.
While molding for 15 days, each group of mice was treated as follows:
the mice in the blank control group and the blank building group (CRS group) are subjected to intragastric administration with 0.1ml/10g of normal saline every day.
CRS + GABA daily: the stomach was perfused with a normal saline containing GABA and spinosin at a ratio of 1:1, the GABA and spinosin were dissolved in a normal saline at a mass ratio of 1:1, and the mice were perfused with a solution of 0.1ml/10g body weight, the perfusion amount of GABA was 20mg/kg body weight, and the perfusion amount of spinosin was 20mg/kg body weight.
The stomach was perfused with normal saline containing GABA and spinosin at a ratio of 1:2 per day, the GABA and spinosin were dissolved in normal saline at a mass ratio of 1:2, and the mice were perfused with 0.1ml of the solution per 10g of body weight, the perfused amount of GABA was 13.3mg/kg of body weight, and the perfused amount of spinosin was 26.7mg/kg of body weight.
The stomach was perfused with normal saline containing GABA and spinosin at a ratio of 1:5 per day, the GABA and spinosin were dissolved in normal saline at a mass ratio of 1:5, and the mice were perfused with 0.1ml of the solution per 10g of body weight, the perfused amount of GABA was 6.7mg/kg of body weight, and the perfused amount of spinosin was 33.3mg/kg of body weight.
The stomach was perfused with normal saline containing GABA and spinosin at a ratio of 1:8 per day, the GABA and spinosin were dissolved in normal saline at a mass ratio of 1:8, and the mice were perfused with 0.1ml of the solution per 10g of body weight, the perfusing amount of GABA was 4.4mg/kg of body weight, and the perfusing amount of spinosin was 35.6mg/kg of body weight.
The stomach was perfused with normal saline containing GABA and spinosin at a ratio of 2:1 per day, the GABA and spinosin were dissolved in normal saline at a mass ratio of 2:1, and the mice were perfused with 0.1ml of the solution per 10g of body weight, the perfusion amount of GABA was 26.7mg/kg of body weight, and the perfusion amount of spinosin was 13.3mg/kg of body weight.
The stomach was perfused with normal saline containing GABA and spinosin at a ratio of 5:1 per day, the GABA and spinosin were dissolved in normal saline at a mass ratio of 5:1, and the mice were perfused with 0.1ml of the solution per 10g of body weight, the perfused amount of GABA was 33.3mg/kg of body weight, and the perfused amount of spinosin was 6.7mg/kg of body weight.
The stomach is irrigated with normal saline containing GABA and spinosin in a group of 8:1 spinosin every day, the GABA and spinosin are dissolved in the normal saline according to the mass ratio of 8:1, the stomach is irrigated to the mice according to 0.1ml solution/10 g of the weight, the lavage amount of the GABA is 35.6mg/kg of the weight, and the lavage amount of the spinosin is 4.4mg/kg of the weight.
CRS +40mg/kg GABA: GABA is dissolved in normal saline to prepare 4g/L solution, and the mouse is subjected to intragastric administration according to the weight of 0.1ml/10 g.
CRS +40mg/kg Spiromone group: the spinosyns were dissolved in physiological saline to make a 4g/L solution, and the mice were gavaged with 0.1ml/10g body weight.
Mouse elevated plus maze experiment:
the next day after 15 days CRS modeling, the mouse elevated plus maze experiment was performed. On the next day after the molding is finished, intragastric administration is carried out on each group of mice according to a grouping treatment mode, and after 60 minutes of intragastric administration, the mice are subjected to an elevated plus maze experiment; at the start of the experiment, mice were placed in the maze from the central compartment towards the open arm, and the activity of each mouse was recorded over 6 minutes using the Any-maze software, with the residence time of the mice in the open arm being observed primarily. After the experiment is completed, the mouse is taken out, the two arms are cleaned, alcohol is sprayed to remove smell, and then the next mouse experiment is continued. The stay time in the open arms was inversely related to the anxiety mood of the mice, and the shorter the stay time, the more severe the anxiety mood of the mice was.
The statistical method comprises the following steps: the residence time of each group of mice is expressed as mean ± SE.
The experimental results are as follows:
the anxiety of the mice was reflected by the retention time, which is shown in table 1 below for each group of mice:
from the above table results, it can be seen that when GABA: in the range of 5:1 to 1:5, the spinosyns show more remarkable anxiolytic effect than the single GABA and the single spinosyns under the condition of the same total dosage, and the ratio of the GABA: the combination of the mass ratios of spinosyns 1:8 and 8:1 does not have this outstanding synergistic effect, so the mass ratio of GABA to spinosyns is preferably 5:1 to 1:5, more preferably 2:1 to 1:2, most preferably 1: 1.
Finally, it should be noted that the above embodiments are only used for illustrating the technical solutions of the present invention and not for limiting, and although the present invention has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions can be made on the technical solutions of the present invention without departing from the spirit and scope of the technical solutions, and all the technical solutions should be covered by the claims of the present invention.
Claims (10)
1. Application of gamma-aminobutyric acid and spinosyn in preparation of products for preventing, relieving or treating anxiety disorders.
2. Use according to claim 1, characterized in that: the mass ratio of the gamma-aminobutyric acid to the spinosin is 1: 0.2 to 5, preferably 1: 0.5-2, more preferably 1: 1.
3. Use according to claim 1 or 2, characterized in that: the content of gamma-aminobutyric acid and the spinosin in the product for preventing, relieving or treating the anxiety disorder is effective amount.
4. Use according to claim 1 or 2, characterized in that: the dosage form of the product for preventing, relieving or treating the anxiety disorder comprises injection, solution, granules, powder, tablets, capsules, pills, oral liquid, suspension, spray or freeze-dried powder injection.
5. Use according to claim 1 or 2, characterized in that: the product for preventing, relieving or treating the anxiety disorder is a medicine or a health-care product.
6. A product for preventing, alleviating or treating anxiety disorders, characterized by: the active ingredients include gamma-aminobutyric acid and spinosin.
7. The article of claim 6, wherein: the mass ratio of the gamma-aminobutyric acid to the spinosin is 1: 0.2 to 5, preferably 1: 0.5-2, more preferably 1: 1.
8. The article of claim 6, wherein: the product is a medicine or a health product.
9. The product of claim 6, 7 or 8, wherein: the content of gamma-aminobutyric acid and the spinosin in the product for preventing, relieving or treating the anxiety disorder is effective amount.
10. The product of claim 5 or 6, wherein: also comprises pharmaceutically acceptable auxiliary materials; preferably, the dosage form comprises injection, solution, granule, powder, tablet, capsule, pill, oral liquid, suspension, spray or lyophilized powder injection.
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