Application of L-tyrosine in medicine for treating obsessive-compulsive disorder

Abstract

The invention discloses application of L-tyrosine (L-tyrosine) in a medicine for treating obsessive-compulsive disorder, the L-tyrosine is used for treating the obsessive-compulsive disorder, and the L-tyrosine is used in a therapeutically effective amount to realize treatment of the obsessive-compulsive disorder. Animal experiments prove that L-tyrosine can influence synthesis of inflammatory factors, neurotrophic factors and dopamine (DA) of neurotransmitters and intestinal barrier function indexes, and then abnormal psychological behavior states related to obsessive-compulsive disorder are greatly relieved. L-tyrosine can improve the compulsive behavior of a model mouse, reduce the level of brain inflammation, recover abnormal changes of neurotrophic factors, repair intestinal barrier function damage and increase secretion of neurotransmitter DA and expression of TH. The invention provides a powerful theoretical basis for seeking the treatment potential of a medicine for obsessive-compulsive disorder, and has higher social significance and potential market value.

Description

Technical Field

[0001] The invention relates to the technical field of biomedicine, and in particular to application of L-tyrosine in drugs for treating obsessive-compulsive disorder. Background Art

[0002] Obsessive-compulsive disorder (OCD) is a chronic, disabling, common mental disorder characterized by recurrent intrusive obsessive thoughts and compulsive behaviors. It has become the fourth most common mental disorder after depression, alcohol / drug abuse, and social phobia. In epidemiological surveys, the prevalence of OCD is very similar among different genders, races, and socioeconomic classes, and there is little evidence that OCD is related to sociodemographics.

[0003] Once OCD becomes severe, it is difficult to relieve itself, and external factors are needed to relieve symptoms. However, the currently recognized first-line treatments for OCD, such as cognitive behavioral therapy (CBT) psychotherapy and selective serotonin reuptake inhibitors (SSRIs) drug therapy, are ineffective, and about 25-40% of patients have no obvious effect on either treatment. Patients who are unresponsive and have no treatment effect can receive other intensive treatment methods, and neurosurgery will be considered in highly refractory and severe cases. This undoubtedly has a great impact on the patient's work, study, social relationships and even daily life, and seriously damages the quality of life of patients and their families. Therefore, it is particularly important to find other alternative treatments.

[0004] L-tyrosine (Tyr), also known as L-β-para-hydroxyphenyl-β-alanine, can be obtained by hydroxylating L-phenylalanine (L-Phe) by phenylalanine hydroxylase (PAH). It is one of the three major aromatic amino acids and plays a very important role in the growth, development and metabolism of humans and animals. Most dopamine (DA) is synthesized directly from tyrosine. First, tyrosine hydroxylase (TH) converts tyrosine into levodopa (L-DOPA), and then DOPA decarboxylase (DDC) converts L-DOPA into DA, which is finally transported to the extracellular space through vesicles to perform its functions. As a major monoamine compound, DA has the functions of regulating motor neurons, spatial memory ability, motivation, excitability, reward and excitement, sexual behavior and nausea. A variety of neurological diseases have been shown to be related to DA signaling defects, such as schizophrenia (SZ), Parkinson's disease (PD), attention deficit and hyperactivity disorder (ADHD), Huntington's disease (HD), addiction, etc. A variety of selective or non-selective dopamine agonists and antagonists have been used clinically to treat SZ, PD, major depressive disorder (MDD), autism spectrum disorder (ASD) and other diseases.

[0005] However, there are currently no studies reporting whether L-tyrosine supplementation can improve OCD-related symptoms. Summary of the invention

[0006] In view of this, the main purpose of the present invention is to provide an application of L-tyrosine in the preparation of a drug for treating obsessive-compulsive disorder, so as to at least solve one of the technical problems in the related art to a certain extent.

[0007] The present invention provides a nutritional supplement that helps alleviate symptoms related to obsessive-compulsive disorder, and provides a new treatment method for people suffering from obsessive-compulsive disorder. It has significant social significance and huge market application value.

[0008] To achieve the above object, the technical solution of the present invention is as follows:

[0009] The invention discloses an application of L-tyrosine in a drug for treating obsessive-compulsive disorder. The L-tyrosine is used for treating obsessive-compulsive disorder, and the L-tyrosine is used in a therapeutically effective amount to achieve the treatment of obsessive-compulsive disorder.

[0010] Furthermore, the obsessive-compulsive disorder includes but is not limited to obsessive-compulsive disorder caused by intestinal immune dysfunction.

[0011] In certain embodiments, the effective amount of L-tyrosine is 140 mg / kg / day-700 mg / kg / day.

[0012] In certain specific embodiments, the L-tyrosine is prepared into a pharmaceutical preparation, and the dosage form of the pharmaceutical preparation includes any pharmaceutically acceptable dosage form.

[0013] Furthermore, the pharmaceutical preparation includes any one of a liquid preparation, a solid preparation, a semisolid preparation and a gas preparation.

[0014] Furthermore, the pharmaceutical preparation also includes any one or more pharmaceutical excipients that are pharmaceutically acceptable.

[0015] In certain specific embodiments, the administration of L-tyrosine includes oral administration, injection administration or spray administration.

[0016] The beneficial effects of the present invention include at least:

[0017] 1) The present invention provides a new use of L-tyrosine for preparing drugs for treating obsessive-compulsive disorder. Animal experiments have shown that L-tyrosine can affect the synthesis of inflammatory factors, neurotrophic factors, neurotransmitter dopamine (DA) and intestinal barrier function indicators, thereby greatly alleviating abnormal psychological and behavioral states associated with obsessive-compulsive disorder. L-tyrosine can improve the compulsive behavior of model mice, reduce brain inflammation levels (Il-6, Tnf-α and Tgf-β), restore abnormal changes in neurotrophic factors (Ngf, Gdnf, Bdnf, Ntf3 and Ntf5), repair intestinal barrier function damage, and increase the secretion of neurotransmitter DA and the expression of TH, providing a new treatment method and theoretical basis for the treatment of obsessive-compulsive disorder.

[0018] 2) This application selects 8-week-old C57BL / 6J healthy male mice weighing 20-22g and subcutaneously injects 1mg / kgQNP in the nape of the neck, and injects once every three days. The control group is given an equal amount of normal saline for a total of 10 injections to construct a subchronic compulsive behavior animal model. After 14 days of intragastric administration of L-tyrosine, it was observed that there were significant improvements in their obsessive-compulsive and anxiety behaviors, and the synthesis of the neurotransmitter dopamine was significantly increased. According to the above experimental results, it is speculated that L-tyrosine may relieve the symptoms of obsessive-compulsive disorder by regulating the synthesis of dopamine. The present invention provides a strong theoretical basis for seeking the therapeutic potential of probiotics for obsessive-compulsive disorder, and has high social significance and potential market value. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 A correlation analysis was performed between the L-tyrosine level described in the present invention and the compulsive behavior of model mice; wherein: (A) the correlation between the cortical L-tyrosine content and the compulsive-like behavior index of mice; (B) the correlation between the cortical L-tyrosine content and the anxiety-like behavior index of mice; (C) the correlation between the plasma L-tyrosine content and the compulsive-like behavior index of mice; and (D) the correlation between the plasma L-tyrosine content and the anxiety-like behavior index of mice.

[0020] Figure 2 The effects of L-tyrosine intervention on the compulsive behavior of model mice described in the present invention; wherein, (A) schematic diagram of the establishment of the L-tyrosine intervention model mice; (B) marble burying test results; (C) Y maze test results; (D) open field test results; (E) elevated plus maze test results; (F) open field test trajectory diagram; (G) elevated plus maze test trajectory diagram.

[0021] Figure 3 The present invention relates to the effects of L-tyrosine intervention on inflammatory factors and neurotrophic factors in the brain of model mice; wherein, (A) inflammatory factor Il-β; (B) inflammatory factor Il-6; (C) inflammatory factor Tnf-α; (D) inflammatory factor Tgf-β; (E) neurotrophic factor Ngf; (F) neurotrophic factor Gdnf; (G) neurotrophic factor Bdnf; (H) neurotrophic factor Ntf3; and (I) neurotrophic factor Ntf5.

[0022] Figure 4 The effects of L-tyrosine intervention on intestinal tight junction proteins in model mice described in the present invention; wherein, (A) Occludin protein expression level after low-dose L-tyrosine intervention; (B) Occludin protein grayscale analysis after low-dose L-tyrosine intervention; (C) Occludin protein expression level after high-dose L-tyrosine intervention; (D) Occludin protein grayscale analysis after high-dose L-tyrosine intervention.

[0023] Figure 5 The present invention relates to the effect of L-tyrosine intervention on the synthesis of the neurotransmitter DA in the cortex of model mice; wherein, (A) the relative expression of Th mRNA after low-dose L-tyrosine intervention; (B) the expression level of TH protein after low-dose L-tyrosine intervention; (C) the grayscale analysis of TH protein after low-dose L-tyrosine intervention; (D) the DA content after low-dose L-tyrosine intervention; (E) the relative expression of Th mRNA after high-dose L-tyrosine intervention; (F) the expression level of TH protein after high-dose L-tyrosine intervention; (G) the grayscale analysis of TH protein after high-dose L-tyrosine intervention; (H) the DA content after high-dose L-tyrosine intervention. DETAILED DESCRIPTION

[0024] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0025] In addition, the technical solutions between the various embodiments of the present invention can be combined with each other, but it must be based on the fact that ordinary technicians in the field can implement it. When the combination of technical solutions is contradictory or cannot be implemented, it should be deemed that such combination of technical solutions does not exist and is not within the scope of protection required by the present invention.

[0026] The experimental methods used in the following examples are conventional methods unless otherwise specified. The materials, reagents, etc. used in the following examples are commercially available unless otherwise specified.

[0027] In the following examples, the configuration method of L-tyrosine used is as follows:

[0028] Weigh L-tyrosine powder and dissolve it in 0.9% saline, then gavage the L-tyrosine gavage group mice at doses of 140 mg / kg / day and 700 mg / kg / day.

[0029] In the following examples, the experimental animals used were: 8-week-old C57BL / 6J healthy male mice weighing 20-22 g, the room temperature was maintained at 23±1°C, the humidity was maintained at 50±10%, the light-dark cycle was 12 hours, and clean food and water were available ad libitum.

[0030] Experimental methods: The experimental animals were randomly divided into three groups: model group (QNP), low-dose L-tyrosine group (L-Tyr), and high-dose L-tyrosine group (H-Tyr). Among them, all three groups of mice were injected with Quinpirole (Quinpirole, QNP) 10 times to construct an obsessive-compulsive disorder model. Two weeks after modeling, they were gavaged once a day with an equal amount of solvent, 140 mg / kg, and 700 mg / kg L-tyrosine. On the 15th day, the mice in each group fasted for 12 hours and behavioral experiments were performed; on the 16th day, the mice in each group fasted for 12 hours, blood was collected from the eye sockets, centrifuged at 3000r / min, and the serum was retained for later use. Brain tissue (cortex) and colon were taken for Western blot, qPCR, HPLC and other tests. All graphs and analyses were performed using GraphPadPrism 8.0. Data were expressed as mean ± standard error (mean ± SEM). One-way analysis of variance (ANOVA) and least significant difference (LSD) multiple test were used to evaluate whether the differences were statistically significant (P < 0.05), *P < 0.05, **P < 0.01, ***P < 0.001.

[0031] Example 1 Correlation analysis between L-tyrosine level and compulsive behavior in model mice

[0032] In this example, general linear regression was used to analyze the correlation between L-tyrosine levels and compulsive behavior in model mice. Figure 1 The results showed that with the decrease of L-tyrosine level, the compulsive burying, compulsive repetition, compulsive inspection and anxiety-like behaviors of QNP-induced model mice increased significantly, indicating that L-tyrosine was negatively correlated with compulsive behavior.

[0033] Example 2 Effect of L-tyrosine intervention on compulsive behavior in QNP-induced model mice

[0034] In this example, an animal model of compulsive behavior was constructed with different doses of L-tyrosine intervention. After 4 weeks of QNP administration and 2 weeks of Tyr gavage, the mice were subjected to behavioral experiments such as marble burying test, Y maze same arm selection test, open field test and elevated plus maze test to observe the effect of L-tyrosine intervention on the compulsive behavior of the model mice, specifically:

[0035] ① Marble burial experiment

[0036] The marble burying (MB) test has been used to evaluate compulsive-like behavior in animals. The experiment followed the protocol described by Deacon et al. Briefly, a clean cage with a length, width and height of 40×24×16 cm was prepared. The bottom of the cage was covered with 5 cm thick wood chip bedding. 18 glass marbles with a diameter of 1.5 cm were placed on the bedding surface, arranged in 6 rows × 3 columns. The mouse was placed individually in the center of the cage and allowed to explore the cage freely without interference under dim light conditions. After 15 minutes, the mouse was removed and the number of marbles buried more than 2 / 3 of the depth was counted. The more marbles buried, the more severe the compulsive burying behavior. The results were expressed as the percentage of the number of marbles buried.

[0037] ②Y-maze

[0038] The reduction of spontaneous alternation behavior in rodents has been proposed as an animal model for persistent symptoms in patients with obsessive-compulsive disorder. The test was performed according to a previously reported procedure. Specifically, in a Y-maze with an arm length of 30 cm, a width of 8 cm, and a height of 15 cm, one of the arms was used as the starting arm. The mouse was placed at the end of the starting arm facing the choice arm. The mouse could freely choose to enter the left or right arm. The selected arm was recorded as the target arm. The mouse was allowed to move freely in the target arm for 5 min. Then the mouse was placed back at the end of the starting arm and observed whether the mouse entered the target arm. This was repeated 20 times, and the number of times the mouse entered the target arm was recorded. The more times the target arm was entered, the more compulsive repetitive behavior was indicated. The results were expressed as the percentage of the number of times the target arm was entered.

[0039] ③Open field experiment

[0040] Excessive inspection is a common and debilitating symptom in patients with obsessive-compulsive disorder. Compulsive inspection behavior in animals can be simulated using a common open field facility (Openfield, OF). The 50×50 cm open field is divided into a grid of 25 virtual rectangular areas, but there are no actual marking lines on the surface of the open field. At the same time, four 4×4×4 cm objects are fixed to the surface of the open field: two in the corners and two near the center. Each mouse is placed individually in the center of the open field and allowed to explore freely for 15 minutes. The trajectory of the mouse is automatically tracked using a camera tracking system. When mice visit 1-2 of the objects excessively frequently and quickly, they are considered to have compulsive inspection behavior.

[0041] ④Elevated plus maze

[0042] Patients with obsessive-compulsive disorder often suffer from anxiety disorders. The elevated plus maze (EPM) is used to test anxiety-related behaviors in rodents. The EPM apparatus is placed 50 cm above the ground and 1 m from the wall, with the closed arm 30 cm long, 5 cm wide, and 15 cm high. The mouse is placed in the EPM apparatus facing the open arm and freely explored for 5 minutes. The trajectory of each animal is automatically tracked using a camera tracking system. The movement distance and duration in the open arm and the reduction in the number of entries into the open arm reflect anxiety-like behavior.

[0043] The behavioral results above are Figure 2 Compared with the mice in the QNP group, the mice in the Tyr group showed a significant reduction in compulsive burying behavior, compulsive repetitive behavior, compulsive checking behavior, and anxiety-like behavior, and both low-dose and high-dose treatments had significant effects. This indicates that L-tyrosine intervention can alleviate the compulsive behavior of QNP-induced model mice.

[0044] Example 3 Effect of L-tyrosine intervention on inflammatory factors and neurotrophic factors in the brain of model mice

[0045] In this example, the qPCR method was used to detect the levels of inflammatory factors and neurotrophic factors in the cerebral cortex tissue of QNP-induced model mice. The results are as follows: Figure 3 As shown:

[0046] Compared with the mice in the QNP group, the mRNA levels of Il-6, Tnf-α, and Tgf-β genes in the cortical tissue of the mice in the Tyr group were reduced to varying degrees ( Figure 3 BD), while the mRNA level of Il-1β gene did not change significantly ( Figure 3 A). In addition, compared with the QNP group mice, the mRNA levels of Ngf and Gdnf genes in the cortex of the Tyr group mice were decreased ( Figure 3 EF), there was no statistical difference in the mRNA levels of Bdnf and Ntf5 genes, and the mRNA level of Ntf3 gene increased in the H-Tyr group ( Figure 3 GI).

[0047] The above results suggest that L-tyrosine intervention can reduce the level of inflammation in the cerebral cortex tissue of QNP-induced model mice and partially restore the changes in neurotrophic factors.

[0048] Example 4 L-tyrosine intervention promotes the expression of tight junction proteins in colon of model mice

[0049] In this example, the Western blot method was used to detect the tight junction protein level in mouse colon and found that L-tyrosine intervention increased the protein level of Occludin ( Figure 4 ), suggesting that L-tyrosine has a protective effect on the intestinal barrier function of mice.

[0050] Example 5 Effect of L-tyrosine intervention on the synthesis of neurotransmitter DA in the cortex of model mice

[0051] In this example, high performance liquid chromatography was used to detect the content of dopamine (DA), a neurotransmitter in the cortex of mice in the Tyr group ( Figure 5 ), qPCR and Western blot were used to detect the expression of tyrosine hydroxylase (TH) in the cortex of Tyr group mice. The results showed that compared with the QNP group mice, the mRNA and protein levels of TH in the cerebral cortex of Tyr group mice increased ( Figure 5 ), the cortical DA content increased.

[0052] The present invention shows through the above experiments that L-tyrosine can alleviate obsessive-compulsive behaviors and anxiety-like behaviors associated with obsessive-compulsive disorder, and this effect may be exerted by regulating the intestinal barrier, brain inflammatory factors, neurotrophic factors and neurotransmitter functions in the gut-brain axis. The present invention provides a strong theoretical basis for seeking the therapeutic potential of tyrosine for obsessive-compulsive disorder, and has high social significance and potential market value.

[0053] It should be noted that, in this article, the terms "include", "comprises" or any other variations thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or device. In the absence of further restrictions, an element defined by the sentence "comprises a ..." does not exclude the existence of other identical elements in the process, method, article or device including the element.

[0054] The serial numbers of the above embodiments of the present invention are only for description and do not represent the advantages or disadvantages of the embodiments.

[0055] The above embodiments are only used to illustrate the technical solutions of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solutions of the present invention can be modified or replaced by equivalents without departing from the purpose and scope of the technical solutions of the present invention, which should be included in the scope of the claims of the present invention.

Claims

1. The application of L-tyrosine in the treatment of obsessive-compulsive disorder.

2. The use according to claim 1, characterized in that: The obsessive-compulsive disorder includes obsessive-compulsive disorder caused by intestinal immune dysfunction.

3. The use according to claim 1, characterized in that: The effective dosage of L-tyrosine is 140 mg / kg / day-700 mg / kg / day.

4. The use according to claim 1, characterized in that: The L-tyrosine is made into a pharmaceutical preparation, and the dosage form of the pharmaceutical preparation includes any pharmaceutically acceptable dosage form.

5. The use according to claim 4, characterized in that: The pharmaceutical preparation includes any one of a liquid preparation, a solid preparation, a semisolid preparation and a gas preparation.

6. The use according to claim 4, characterized in that: The pharmaceutical preparation also includes any one or more pharmaceutical excipients that are acceptable in pharmaceutical science.

7. The use according to any one of claims 1 to 6, characterized in that: The administration method of L-tyrosine includes oral administration, injection administration or spray administration.

Images