GLP-1 receptor agonist or pharmaceutical composition thereof for improving or alleviating dopamine dependence

Abstract

The invention provides a GLP-1 receptor agonist for improving or relieving dopamine dependence, and particularly relates to application of semeglutide in treating, improving or relieving dopamine addiction, the application dosage form can be a solid preparation or a liquid preparation, and the invention discloses a prescription composition and a preparation method of a related preparation composition. And the pharmaceutical composition provided by the invention has a good effect when being used for treating behaviors closely associated with dopamine, such as eating addiction, drinking addiction, coffee addiction, drug addiction and the like.

Description

Technical Field

[0001] The present invention relates to the use of a GLP-1 receptor agonist or a pharmaceutical composition thereof for improving or alleviating dopamine dependence, and specifically relates to the use of semaglutide or a pharmaceutical composition thereof for improving or alleviating dopamine dependence and inhibiting negative emotional states resulting from dopamine addiction. Background Art

[0002] Addiction has always been a serious problem. According to its development process, addiction can be divided into three stages, namely abuse and addiction, withdrawal and depression, and possession and craving. With the continuous cycle of these three stages, the symptoms of addicts will continue to worsen without treatment intervention.

[0003] In the first stage, addictive substances cause the reward area in the brain to secrete a large amount of dopamine. The sharp rise in dopamine levels brings pleasure to the individual, and the brain associates the pleasure with the external environment before the pleasure occurs (i.e., the cause of addiction). With the continuous repetition of this reward stimulus, dopamine cells no longer respond to the reward itself, but instead discharge in anticipation of the stimulus. However, unlike natural rewards, dopamine cells in the brain do not stop firing after repeated intake of addictive substances, but instead continuously increase the level of dopamine secretion. In short, changes in dopamine levels are the main reason for addicts to have impulsive behavior. In the second stage, the dopamine system in the brain has obvious functional disorders. An important consequence of this disorder is that healthy reward stimuli cannot produce the normal driving force level before addiction occurs. After conditioned learning of the dopamine system, the addict's reward and motivation system has been changed, and it only focuses on the stimuli related to the addictive substance. In addition, repeated exposure to the stimulus will also cause adaptive changes in the brain circuits in the brain, including the amygdala, thus producing the withdrawal & depression stage. In this stage, individuals become more reactive to stress, which amplifies negative emotions, such as anxiety and other negative emotional states. The imbalance of homeostasis and the amplified negative emotions drive individuals to take actions to escape the discomfort after the disappearance of the addictive substance, and the solution is often to use the addictive substance again, thus creating a vicious cycle of addiction. In the third stage, in addition to the changes in the above-mentioned reward and emotional circuits, the deepening of addiction is also accompanied by damage to the function of the prefrontal cortex area. The disorder of dopamine signals in the prefrontal region seriously impairs various executive functions, such as self-regulation, decision-making, choice flexibility and action initiation, salience allocation, and error monitoring. The disorder of dopamine signals weakens the individual's resistance to the impulse to consume addictive substances and also hinders their decision to stop taking them. Therefore, damage to the prefrontal regulatory circuit is one of the root causes of the deterioration of impulsive drug intake behavior after addiction.

[0004] Alcohol is a well-known addictive substance, and alcohol addiction is the third most common global public health problem. According to the WHO report, from 1990 to 2020, drinking has risen from sixth to third in the list of all disease risk factors, second only to hypertension and smoking. Drinking causes about 4 million deaths worldwide each year. Alcohol addiction is also known as alcohol use disorder (AUD). Alcohol addiction is characterized by compulsive seeking and intake of alcohol, loss of control over intake, and negative emotional states such as anxiety during withdrawal. The process is generally divided into three stages: trying to drink; losing control of drinking, that is, increasing drinking; and compulsive seeking and intake of alcohol. The principle is that the mesolimbic dopamine system positively reinforces alcohol addiction. Alcohol and other drugs of abuse (morphine, cocaine, ketamine, etc.) addiction are all reinforced by activating the mesolimbic dopamine system. The mesolimbic dopamine system consists of the ventral tegmental area (VTA) and the nucleus accumbens (NAC). The mechanism of strengthening alcohol addiction is: alcohol stimulates dopamine neurons in the VTA to release dopamine, and at the same time, alcohol activates the μ opioid receptors on the cell membrane of γ-aminobutyric acid neurons in the VTA to release the inhibition of dopamine neurons, thereby releasing dopamine. In addition to delivering information of excitement and joy, dopamine can also be addictive. It can be said that people's addiction to some things is mainly due to dopamine secretion. Dopamine acts on dopamine D1 and D2 receptors in the prefrontal cortex. When ingesting addictive substances such as drinking, it will simulate the high concentration of dopamine produced by staged discharge, activate D1 and D2 receptors to make the body feel pleasant, and strengthen alcohol addiction. There are currently no treatment options that can be used to actively improve or alleviate alcohol dependence. According to a survey report by the World Health Organization, one in every 100 Chinese people over the age of 15 has the potential to or has already suffered from "alcohol addiction". At the same time, more than hundreds of thousands of people in my country die from alcohol poisoning due to symptoms of alcohol dependence every year, accounting for 2.3% of the mortality rate, and more than two million people are disabled, accounting for 4.0% of the total disability rate. As of today, the death and disability rates due to alcohol dependence are still rising.

[0005] Chinese patent 2018800585280 discloses that GLP-1 receptor agonists have the potential to treat obesity, but it is only described through primary trials, and the detailed treatment principle is not disclosed, making it difficult for the public to trust its test conclusions. To date, there is no data to confirm and elaborate on the use of GLP-1 receptor agonists in affecting addiction, so determining whether GLP-1 receptor agonists have an effect on dopamine addiction and the negative states caused by addiction is an important issue to be resolved. Summary of the invention

[0006] During the experiment, the inventors surprisingly found that GLP-1 receptor agonists have the ability to improve or alleviate dopamine dependence, specifically the ability of GLP-1 receptor agonists to regulate negative emotions after addiction. The present invention discloses a method for improving or alleviating dopamine dependence and inhibiting negative states (such as anxiety) produced after addiction, which comprises administering a GLP-1 receptor agonist or a pharmaceutical composition thereof.

[0007] In some embodiments, the present invention relates to a method for improving or alleviating dopamine dependence, wherein the method comprises administering a GLP-1 receptor agonist to a subject in need thereof and the subject suffers from metabolic syndrome. In some embodiments, the present invention relates to a method for improving or alleviating dopamine dependence, wherein the method comprises administering a GLP-1 receptor agonist to a human subject in need thereof and the subject suffers from one or more indications selected from prediabetes, diabetes, cardiovascular disease, obesity and hypertension. In some embodiments, the present invention relates to a method for improving or alleviating dopamine dependence, wherein the method comprises administering semaglutide to a subject in need thereof.

[0008] In some embodiments, the present invention relates to a method for improving or alleviating the risk of dopamine dependence, wherein the method comprises administering a GLP-1 receptor agonist to a subject in need thereof and the subject suffers from metabolic syndrome. In some embodiments, the present invention relates to a method for improving or alleviating the risk of dopamine dependence, wherein the method comprises administering a GLP-1 receptor agonist to a subject in need thereof and the subject suffers from one or more indications selected from prediabetes, diabetes, cardiovascular disease, obesity and hypertension. In some embodiments, the present invention relates to a method for improving or alleviating the risk of dopamine dependence, wherein the method comprises administering semaglutide to a subject in need thereof.

[0009] There are many types of addiction. The addiction described in the present invention refers to addiction caused by dopamine dependence. Representative addictive substances include food, alcohol, caffeine, marijuana, various hallucinogens, inhalants, opium, tranquilizers, sleeping pills, stimulants, cigarettes, etc. The representative manifestations of substance addiction are A. Impulsive seeking and use of drugs; B. Uncontrolled use / intake behavior; C. A large number of negative emotional states after withdrawal from drugs, such as anxiety and other negative emotional states. For healthy individuals, impulsive traits are a personality factor that is highly correlated with addictive behavior. Impulsiveness refers to a personality trait that lacks planning when taking actions and hardly considers the possible consequences of such behavior. Direct evidence from animal models shows that impulsivity is directly related to addictive behavior, and human evidence from personality, behavior, and neurophysiology shows that impulsivity can be summarized by a two-factor model. The first is a bottom-up process, which corresponds to the enhanced motivational activity of the amygdala, ventral striatum and other brain regions in response to reward stimuli. Some researchers use terms such as "stimulation seeking" and "oversensitivity to rewards" to describe this process. The second is a reduced top-down behavioral control ability, and the corresponding neural mechanism is abnormalities in the ventromedial and dorsolateral prefrontal cortex, anterior cingulate, and insula in the cortical system. Commonly used adjectives include "weakened self-regulation", "impaired response inhibition", and "high impulsivity".

[0010] Alcohol addiction is a representative of dopamine dependence. The drinking characteristics of alcohol addicts are mainly manifested in long-term heavy drinking. Studies have shown that the transition from occasional moderate drinking to frequent and uncontrolled drinking is mainly related to the activation of the reward system. Alcohol, as a reward substance, can produce excitement and pleasure after an individual consumes it, thereby triggering a reward effect and generating craving and dependence on it. The activation of the reward system is mainly regulated by the mesolimbic dopamine system. The ventral tegmental area (VTA) of the midbrain contains a large number of dopamine (DA) neurons, which can project to other different brain regions, including the nucleus accumbens (NAc), central amygdala (CeA), medial prefrontal cortex (mPFC) and hippocampus (HP), thereby regulating a series of behaviors, such as learning, memory, anxiety, depression, etc. Glucagon-like peptide-1 (GLP-1) is an incretin hormone and neuropeptide that was originally found to be synthesized in the intestine. It was later found that it can cross the blood-brain barrier and enter the central nervous system (CNS), and can also be produced by neurons and microglia in the CNS. Numerous studies have shown that GLP-1 plays an important role in regulating food intake in the brainstem and hypothalamus. Studies have found that GLP-1 can also regulate food reward behavior through reward circuits (including VTA and NAc).

[0011] In some embodiments, the term "treatment" as used herein includes prevention, delay, relief, reduction of risk of occurrence, improvement or curative treatment for the mentioned medical indications. Treatment can be symptomatic treatment or treatment to improve the disease. In some embodiments, the term treatment as used herein refers to the prevention of the mentioned medical indications. In some embodiments, the term treatment as used herein refers to delaying (e.g., delaying the onset) the mentioned medical indications. In some embodiments, the term treatment as used herein refers to reducing the risk of the mentioned medical indications. In some embodiments, the term treatment as used herein refers to improving the mentioned medical indications. In some embodiments, the term treatment as used herein refers to curatively treating the mentioned medical indications.

[0012] In some embodiments, the subject to which the GLP-1 receptor agonist of the present invention is to be administered can be a human, such as an adult (also referred to as an adult). In some embodiments, the subject suffers from metabolic syndrome. As used herein, the term "metabolic syndrome" refers to one or more indications selected from prediabetes, diabetes, cardiovascular disease, obesity and hypertension. In some embodiments, "metabolic syndrome" refers to at least two indications selected from prediabetes, diabetes, cardiovascular disease, obesity and hypertension. In some embodiments, "metabolic syndrome" refers to at least three indications selected from prediabetes, diabetes, cardiovascular disease, obesity and hypertension. In some embodiments, the subject suffers from prediabetes. In some embodiments, the subject suffers from diabetes. In some embodiments, diabetes is type 2 diabetes. In some embodiments, the subject suffers from obesity. In some embodiments, the subject suffers from cardiovascular disease. In some embodiments, the subject suffers from obesity. In some embodiments, obesity is a BMI of at least 25kg / m2, such as at least 27kg / m2 or at least 30kg / m2. In some embodiments, the subject suffers from hypertension. In some embodiments, cardiovascular disease includes one or more of coronary artery disease (such as angina and myocardial infarction), stroke, heart failure, hypertensive heart disease, rheumatic heart disease, cardiomyopathy, arrhythmia, congenital heart disease, valvular heart disease, carditis, aortic aneurysm, peripheral arterial disease, thromboembolic disease and venous thrombosis. In some embodiments, the subject has previously been administered insulin.

[0013] In some embodiments, the term "GLP-1 receptor agonist" as used in the present invention refers to a compound that fully or partially activates the human GLP-1 receptor. In some embodiments, the GLP-1 receptor agonist used in the present invention is an acylated GLP-1 receptor agonist. The term "acylation" used in connection with the GLP-1 receptor agonist means that the GLP-1 receptor agonist is covalently linked to at least one substituent comprising a lipophilic portion such as a fatty acid or a fatty diacid. In some embodiments, the substituent comprises a fatty acid or a fatty diacid. In some embodiments, the term GLP-1 receptor agonist and the specific GLP-1 receptor agonists described herein also encompass salt forms thereof. Therefore, the GLP-1 receptor agonist should exhibit "GLP-1 activity", which refers to the ability of the compound (i.e., the GLP-1 receptor agonist) to bind to the GLP-1 receptor and initiate a signal transduction pathway, thereby resulting in an insulinotropic effect or other physiological effects known in the art.

[0014] In some embodiments, the methods of the present invention include the GLP-1 receptor agonist semaglutide. Semaglutide is N-epsilon26-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(17-carboxyheptadecanoyl-amino)butyrylamino]ethoxy}ethoxy)acetylamino]ethoxy}ethoxy)acetyl][Aib8,Arg34]GLP-1-(7-37). In some embodiments, semaglutide is administered once a day or less frequently, for example once a day or once a week. In some embodiments, the GLP-1 receptor agonist is administered at any time of the day. In some embodiments, the methods of the present invention last for at least about 48 weeks, such as at least about 1 year.

[0015] In some embodiments, semaglutide is the only GLP-1 receptor agonist administered in the methods of the present invention. In some embodiments, semaglutide is the only active ingredient administered in the methods of the present invention. In some embodiments, semaglutide is the only active ingredient administered in the methods of the present invention. In some embodiments, the methods of the present invention include administering to a subject one or more additional active ingredients that are not GLP-1 receptor agonists.

[0016] In some embodiments, the GLP-1 receptor agonist can be administered in the form of a pharmaceutical composition. The pharmaceutical composition can be in liquid or solid form.

[0017] In some embodiments, the pharmaceutical composition formed by the GLP-1 receptor agonist is administered parenterally, and the pharmaceutical composition may contain a GLP-1 receptor agonist at a concentration of 0.1 mg / ml to 100 mg / ml. In some embodiments, the pharmaceutical composition contains 0.01-50 mg / ml, or 0.01-20 mg / ml, or 0.01-10 mg / ml of GLP-1 receptor agonist. In some embodiments, the pharmaceutical composition contains 0.1-20 mg / ml of GLP-1 receptor agonist.

[0018] In some embodiments, the pharmaceutical composition may further comprise one or more pharmaceutically acceptable excipients, such as excipients selected from a buffer system, a preservative, a tonicity agent, a chelating agent, a stabilizer, and a surfactant. In some embodiments, the pharmaceutical composition comprises one or more pharmaceutically acceptable excipients, such as one or more selected from a buffer, an isotonic agent, and a preservative. The formulation of pharmaceutical active ingredients with various excipients is known in the art, and the term "excipient" refers to any component other than an active therapeutic ingredient such as a GLP-1 receptor agonist. Excipients may be inert substances, inactive substances, and / or non-pharmaceutically active substances.

[0019] In some embodiments, the pH of the pharmaceutical composition is in the range of 7.0-10.0, such as 7.0 to 9.5 or 7.2 to 9.5. In some embodiments, the pH of the pharmaceutical composition is in the range of 7.0-8.5, such as 7.0 to 7.8 or 7.8 to 8.2. In some embodiments, the pH of the pharmaceutical composition is 7.4. In some embodiments, the pH of the pharmaceutical composition is 8.15. In some embodiments, the pharmaceutical composition comprises a phosphate buffer, such as a sodium phosphate buffer, for example, disodium phosphate. In some embodiments, the pharmaceutical composition comprises an isotonic agent, such as propylene glycol. In some embodiments, the pharmaceutical composition comprises a preservative, such as phenol.

[0020] In some embodiments, the pharmaceutical composition can be in the form of a solution or suspension. In some embodiments, the pharmaceutical composition is an aqueous composition, such as an aqueous solution or an aqueous suspension. The term "aqueous composition" is defined as a composition comprising at least 50% w / w water. Similarly, the term "aqueous solution" is defined as a solution comprising at least 50% w / w water, and the term "aqueous suspension" is defined as a suspension comprising at least 50% w / w water. Aqueous compositions may comprise at least 50% w / w water, or at least 60%, 70%, 80% or even at least 90% w / w water.

[0021] In some embodiments, the GLP-1 receptor agonist is administered in the form of a pharmaceutical composition comprising about 0.1-20 mg / ml of the GLP-1 receptor agonist, about 2-15 mM phosphate buffer, about 2-25 mg / ml of propylene glycol, and a pH in the range of 7.0-9.0. In some embodiments, the GLP-1 receptor agonist is administered in the form of a pharmaceutical composition comprising about 0.1-20 mg / ml of the GLP-1 receptor agonist, about 2-15 mM phosphate buffer, about 2-25 mg / ml of propylene glycol, about 1-18 mg / ml of phenol, and a pH in the range of 7.0-9.0. In some embodiments, the GLP-1 receptor agonist (e.g., semaglutide) is administered in the form of a pharmaceutical composition comprising about 1.34 mg / ml of the GLP-1 receptor agonist (e.g., semaglutide), about 1.42 mg / ml of disodium phosphate dihydrate, about 14.0 mg / ml of propylene glycol, about 5.5 mg / ml of phenol, and a pH of about 7.4. In some embodiments, the GLP-1 receptor agonist (e.g., semaglutide) is administered in the form of a pharmaceutical composition comprising 1.34 mg / ml of the GLP-1 receptor agonist, 1.42 mg / ml of disodium phosphate dihydrate, 14.0 mg / ml of propylene glycol, 5.5 mg / ml of phenol, and a pH of 7.4. In some embodiments, the GLP-1 receptor agonist (e.g., liraglutide) is administered in the form of a pharmaceutical composition comprising a GLP-1 receptor agonist (e.g., 3-8 mg / ml), disodium phosphate dihydrate, propylene glycol, phenol, and a pH of about 8.0-8.3. In some embodiments, the GLP-1 receptor agonist (e.g., liraglutide) is administered in the form of a pharmaceutical composition comprising a GLP-1 receptor agonist (e.g., about 6.0 mg / ml), disodium phosphate dihydrate, propylene glycol, phenol, and a pH of about 8.15.

[0022] In some embodiments, the pharmaceutical composition formed from the GLP-1 receptor agonist is a solid composition.

[0023] In some embodiments, the solid composition can be a solid composition suitable for administration by oral route, and in some embodiments, the solid composition comprises at least one pharmaceutically acceptable excipient. As used herein, the term "excipient" refers to any component except active therapeutic ingredients or active pharmaceutical ingredients (API). Excipients can be pharmaceutically inert substances, inactive substances and / or therapeutically or medically inactive substances. Excipients can be used for a variety of purposes, such as as carriers, vehicles, fillers, adhesives, lubricants, glidants, disintegrants, flow control agents, crystallization inhibitors, solubilizers, stabilizers, colorants, flavorings, surfactants, emulsifiers or combinations thereof, and / or for improving the administration and / or absorption of therapeutically active substances or active pharmaceutical ingredients. The amount of each excipient used can vary within the conventional range of this area. In some embodiments, the excipients may be selected from binders, such as polyvinyl pyrrolidone (povidone), etc.; fillers, such as cellulose powder, microcrystalline cellulose, cellulose derivatives, such as hydroxymethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose and hydroxypropyl methyl cellulose, calcium hydrogen phosphate, corn starch, pregelatinized starch, etc.; lubricants and / or glidants, such as stearic acid, magnesium stearate, sodium stearyl fumarate, glyceryl tribehenate, etc.; flow control agents, such as colloidal silicon dioxide, talc, etc.; crystals; Inhibitors, such as povidone, etc.; solubilizers, such as Pluronic, povidone, etc.; colorants, including dyes and pigments, such as red iron oxide or yellow iron oxide, titanium dioxide, talc, etc.; pH control agents, such as citric acid, tartaric acid, fumaric acid, sodium citrate, calcium hydrogen phosphate, disodium hydrogen phosphate, etc.; surfactants and emulsifiers, such as Pluronic, polyethylene glycol, sodium carboxymethyl cellulose, polyethoxylated and hydrogenated castor oil, etc.; and mixtures of two or more of these excipients and / or adjuvants.

[0024] In some embodiments, the solid composition may include a binder such as povidone; starch; cellulose and its derivatives such as microcrystalline cellulose, hydroxypropyl cellulose, hydroxyethyl cellulose and hydroxypropyl methylcellulose; sucrose; dextrose; corn syrup; polysaccharides; and gelatin. The binder may be selected from a dry granulation binder and / or a wet granulation binder. Suitable dry granulation binders are, for example, cellulose powder and microcrystalline cellulose such as Avicel PH 102 and Avicel PH200. In some embodiments, the solid composition includes Avicel such as Aavicel PH 102. Suitable binders for wet granulation or dry granulation are corn starch, polyvinyl pyrrolidone (povidone), vinyl pyrrolidone-vinyl acetate copolymer (copovidone) and cellulose derivatives such as hydroxymethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose and hydroxypropyl methylcellulose. In some embodiments, the solid composition includes povidone.

[0025] In some embodiments, the solid composition comprises a filler, which may be selected from lactose, mannitol, erythritol, sucrose, sorbitol, calcium phosphate such as dibasic calcium phosphate, microcrystalline cellulose, powdered cellulose, confectioner's sugar, compressible sugar, dextrates, dextrin and dextrose. In some embodiments, the solid composition comprises microcrystalline cellulose, such as Avicel PH102 or Avicel PH 200.

[0026] In some embodiments, the solid composition comprises a lubricant and / or a glidant. In some embodiments, the composition comprises a lubricant and / or a glidant, such as talc, magnesium stearate, calcium stearate, zinc stearate, glyceryl behenate, glyceryl dibehenate, behenyl polyoxyethylene-8 glyceride, polyethylene oxide polymer, sodium lauryl sulfate, magnesium lauryl sulfate, sodium oleate, sodium stearyl fumarate, stearic acid, hydrogenated vegetable oil, silicon dioxide and / or polyethylene glycol, etc. In some embodiments, the solid composition comprises magnesium stearate or glyceryl dibehenate.

[0027] In some embodiments, the solid composition comprises a disintegrant, such as sodium starch glycolate, polacrilin potassium, sodium starch glycolate, crospovidone, cross-linked carboxymethylcellulose, sodium carboxymethylcellulose, or dried corn starch. The solid composition may comprise one or more surfactants, such as one surfactant, at least one surfactant, or two different surfactants. The term "surfactant" refers to any molecule or ion consisting of a water-soluble (hydrophilic) part and a fat-soluble (lipophilic) part. The surfactant can be selected, for example, from anionic surfactants, cationic surfactants, nonionic surfactants, and / or zwitterionic surfactants.

[0028] The solid composition may further comprise a delivery agent or an absorption enhancer, which, for the present invention, is an adjuvant capable of increasing the oral exposure of the GLP-1 receptor agonist. The delivery agent may be a salt of N-(8-(2-hydroxybenzoyl)amino)caprylic acid (referred to herein as a salt of NAC), which contains an anion N-(8-(2-hydroxybenzoyl)amino)caprylic acid group.

[0029] In some embodiments, the salt of N-(8-(2-hydroxybenzoyl)amino)caprylic acid comprises one monovalent cation, two monovalent cations, or one divalent cation. In some embodiments, the salt of N-(8-(2-hydroxybenzoyl)amino)caprylic acid is selected from the sodium salt, potassium salt, and / or calcium salt of N-(8-(2-hydroxybenzoyl)amino)caprylic acid. In some embodiments, the salt of N-(8-(2-hydroxybenzoyl)amino)caprylic acid is selected from the sodium salt, potassium salt, and / or ammonium salt. In some embodiments, the salt of N-(8-(2-hydroxybenzoyl)amino)caprylic acid is a sodium salt or a potassium salt. In some embodiments, the salt of N-(8-(2-hydroxybenzoyl)amino)caprylic acid may be crystalline and / or amorphous. In some embodiments, the delivery agent includes an anhydrate, monohydrate, dihydrate, trihydrate, solvate or one-third hydrate of N-(8-(2-hydroxybenzoyl)amino)caprylic acid salt, and combinations thereof. In some embodiments, the delivery agent is sodium N-(8-(2-hydroxybenzoyl)amino)caprylate (referred to herein as SNAC). In some embodiments, the delivery agent is sodium 8-(salicyloylamino)caprylate.

[0030] In some embodiments, the composition for use in the present invention is in the form of a solid composition for oral administration, such as a tablet.

[0031] In some embodiments, the solid composition comprises 0.1–50 mg, such as 0.5 to 40 mg or 1-30 mg of a GLP-1 receptor agonist. In some embodiments, the solid composition comprises 2-20 mg, such as 3-18 mg or 5-15 mg of a GLP-1 receptor agonist. In some embodiments, the solid composition comprises about 3 mg, such as about 7 mg or about 14 mg of a GLP-1 receptor agonist.

[0032] In some embodiments, at least 30% (w / w) of the solid composition (e.g., tablet) is a salt of N-(8-(2-hydroxybenzoyl)amino)caprylic acid. In some embodiments, at least 50% (w / w) of the solid composition (e.g., tablet) is a salt of N-(8-(2-hydroxybenzoyl)amino)caprylic acid. In some embodiments, the amount of N-(8-(2-hydroxybenzoyl)amino)caprylic acid salt in each dosage unit of the composition is in the range of 0.20-.5mmol, 0.25-1.0mmol, 0.30-0.75mmol, or such as 0.45-0.65mmol. In some embodiments, the amount of SNAC in the composition is in the range of 75-600mg. In some embodiments, the amount of SNAC in the composition is in the range of 75-400mg, such as 80-350mg, such as about 100mg to about 300mg per dosage unit.

[0033] In some embodiments, the solid composition comprises a lubricant, such as magnesium stearate.In some embodiments, the unit dose of the solid composition comprises: 0.1-50 mg of a GLP-1 receptor agonist, 25-600 mg of a salt of N-(8-(2-hydroxybenzoyl)amino)caprylic acid (NAC) (such as sodium salt of NAC (SNAC)), and 0-25 mg of a lubricant.

[0034] In some embodiments, the GLP-1 receptor agonist can be administered by parenteral administration, such as subcutaneous injection. The GLP-1 receptor agonist can be administered using a pen injector, such as a 3 ml disposable pen injector. In some embodiments, the dose of the GLP-1 receptor agonist is in the range of 0.1 to 5.0 mg, such as in the range of 0.1 to 3.0 mg. In some embodiments, the daily dose of the GLP-1 receptor agonist is selected from 0.5 and 1.0 mg.

[0035] In some embodiments, the GLP-1 receptor agonist can be administered orally, for example in the form of tablets, coated tablets, sachets or capsules such as hard or soft gelatin capsules, and all such compositions are considered solid oral dosage forms. Oral administration can be once daily administration. In some embodiments, the dose of the GLP-1 receptor agonist is in the range of 1 to 30 mg, such as 2 to 20 mg or 3 to 15 mg. In some embodiments, the dose of the GLP-1 receptor agonist is 3, 7 or 14 mg. The composition can be in the form of a dosage unit such as a tablet. In some embodiments, the weight of the unit dose is in the range of 50 mg to 1000 mg, such as 50-750 mg or, for example, in the range of about 100-500 mg. In some embodiments, In the embodiment, the weight of the dosage unit is in the range of 75 mg to 350 mg, such as 50-300 mg or 100-400 mg. Tablets for oral administration may contain 30% (w / w) of a salt of N-(8-(2-hydroxybenzoyl)amino)caprylic acid, such as the sodium salt of N-(8-(2-hydroxybenzoyl)amino)caprylic acid (SNAC). In some embodiments, the composition may be granulated before being compressed into a tablet. The composition may comprise a granular portion and / or an extragranular portion, wherein the granular portion has been granulated and the extragranular portion has been added after granulation. The GLP-1 receptor agonist may be contained in the granular portion or the extragranular portion. In some embodiments, the extragranular portion comprises a GLP-1 receptor agonist. In an embodiment, the extragranular portion may further comprise a lubricant and / or a glidant. In an embodiment, the granular portion may comprise a lubricant and / or a glidant. In an embodiment, the granular portion and the extragranular portion comprise a lubricant and / or a glidant.

[0036] In some embodiments, the GLP-1 receptor agonist is the only active pharmaceutical ingredient administered according to the present invention. In some embodiments, the GLP-1 receptor agonist is combined with one or more other active pharmaceutical ingredients in the present invention. In some embodiments, the term "long-term treatment" as used herein for GLP-1 receptor agonists refers to administration in an amount and frequency that provides a therapeutic effect. In some embodiments, the term "long-term treatment" as used herein for GLP-1 receptor agonists refers to administration of 0.1-3.0 mg, such as 0.5 or 1.0 mg GLP-1 receptor agonist (e.g., semaglutide) once a week. In some embodiments, the term "long-term treatment" as used herein for GLP-1 receptor agonists refers to administration of 0.05-0.3 mg, such as 0.05, 0.1, 0.2 or 0.3 mg GLP-1 receptor agonist (e.g., semaglutide) once a day. In some embodiments, the term "long-term treatment" as used herein with respect to a GLP-1 receptor agonist refers to once daily administration of 0.1-6 mg, such as 0.5-4 mg of a GLP-1 receptor agonist (e.g., semaglutide). As used herein, the term "long-term treatment" may refer to long-term (e.g., at least 2 years or at least 5 years) administration of a drug according to a prescribed dosage regimen (e.g., once weekly administration), wherein up to 10%, such as up to 5%, of the dose may be missed; provided that the missed dose does not exceed 10 consecutive times.

[0037] Unless otherwise stated, the ranges herein include their endpoints. In some embodiments, the term "a" means "one (kind) or more (kind)". In some embodiments, and unless otherwise stated in this specification, the terms presented in the singular also include plural cases. In this article, the term "about" refers to the value referred to ± 10%, and includes the value.

[0038] Non-limiting embodiments of the present invention include: A GLP-1 receptor agonist for improving or alleviating dopamine dependence, characterized in that the GLP-1 receptor agonist is the only active pharmaceutical ingredient or the only GLP-1 receptor agonist administered to the subject.

[0039] In some embodiments, a GLP-1 receptor agonist for improving or alleviating dopamine dependence, wherein the GLP-1 receptor agonist is liraglutide, dulaglutide, semaglutide, or telpotide.

[0040] In some embodiments, a GLP-1 receptor agonist is used to improve or alleviate dopamine dependence, wherein the GLP-1 receptor agonist is semaglutide.

[0041] In some embodiments, a GLP-1 receptor agonist for improving or alleviating dopamine dependence, wherein the GLP-1 receptor agonist is semaglutide, further comprises another therapeutic agent in addition to semaglutide, and the other therapeutic agent may be liraglutide, dulaglutide, retatrutide, or telpotide.

[0042] In some embodiments, a pharmaceutical composition consisting of a GLP-1 receptor agonist and a pharmaceutically acceptable excipient is provided for improving or alleviating dopamine dependence.

[0043] In some embodiments, a pharmaceutical composition consisting of a GLP-1 receptor agonist and a pharmaceutically acceptable excipient for improving or alleviating dopamine dependence, wherein the GLP-1 receptor agonist is the only active pharmaceutical ingredient or the only GLP-1 receptor agonist administered to the subject.

[0044] In some embodiments, a pharmaceutical composition consisting of a GLP-1 receptor agonist and a pharmaceutically acceptable excipient for improving or alleviating dopamine dependence, wherein the GLP-1 receptor agonist is semaglutide.

[0045] In some embodiments, a pharmaceutical composition consisting of a GLP-1 receptor agonist and a pharmaceutically acceptable excipient for improving or alleviating dopamine dependence, wherein the pharmaceutical composition is administered subcutaneously in the form of a solution containing at least 90% water and having a pH in the range of 7.0-9.0.

[0046] In some embodiments, a pharmaceutical composition consisting of a GLP-1 receptor agonist and a pharmaceutically acceptable excipient for improving or alleviating dopamine dependence, wherein the formulation excipient includes propylene glycol, and the concentration of the propylene glycol is about 1 mg / ml to about 100 mg / ml.

[0047] In some embodiments, a pharmaceutical composition consisting of a GLP-1 receptor agonist and a pharmaceutically acceptable excipient for improving or alleviating dopamine dependence, wherein the concentration of the formulation excipient propylene glycol is about 1 mg / ml to about 50 mg / ml.

[0048] In some embodiments, a pharmaceutical composition consisting of a GLP-1 receptor agonist and a pharmaceutically acceptable excipient for improving or alleviating dopamine dependence, wherein the concentration of the formulation excipient propylene glycol is about 5 mg / ml to about 25 mg / ml.

[0049] In some embodiments, a pharmaceutical composition consisting of a GLP-1 receptor agonist and a pharmaceutically acceptable excipient for improving or alleviating dopamine dependence, the formulation excipient further comprises a preservative, wherein the preservative is present at a concentration of 0.1 mg / ml to 20 mg / ml.

[0050] In some embodiments, a pharmaceutical composition consisting of a GLP-1 receptor agonist and a pharmaceutically acceptable excipient for improving or alleviating dopamine dependence, the formulation excipient further includes a buffer, wherein the buffer is selected from glycylglycine, L-histidine, Hepes, N-bicine and disodium phosphate dihydrate.

[0051] In some embodiments, a method for preparing a pharmaceutical composition consisting of semaglutide and pharmaceutically acceptable excipients having the function of improving or alleviating dopamine dependence comprises: a) dissolving a preservative, propylene glycol and a buffer in water to prepare a first solution; b) dissolving semaglutide in water to prepare a second solution; c) mixing the first and second solutions; and d) adjusting the pH of the mixture in c) to a pH of about 7.0 to about 9.0.

[0052] In some embodiments, a pharmaceutical composition consisting of a GLP-1 receptor agonist and a pharmaceutically acceptable excipient for improving or alleviating dopamine dependence is administered orally.

[0053] In some embodiments, a pharmaceutical composition consisting of a GLP-1 receptor agonist and a pharmaceutically acceptable excipient for improving or alleviating dopamine dependence is administered in the form of a tablet, which contains at least 30% (w / w) of N-(8-(2-hydroxybenzoyl)amino)caprylic acid salt.

[0054] In some embodiments, an oral pharmaceutical composition consisting of a GLP-1 receptor agonist and a pharmaceutically acceptable excipient for improving or alleviating dopamine dependence comprises at least 60% (w / w) of a delivery agent, less than 10% (w / w) of a binder, 5-40% (w / w) of a filler, less than 10% (w / w) of a lubricant or a glidant, and at least 70% (w / w) or at least 75% (w / w) of a delivery agent.

[0055] In some embodiments, the oral composition comprises 0.1-10% (w / w), such as 0.2-4% (w / w) or 0.5-3% (w / w) of a binder. In some embodiments, the composition comprises 1% (w / w) or 2% (w / w) of a binder. The composition may comprise a binder, such as povidone; starch; cellulose and its derivatives, such as microcrystalline cellulose, hydroxypropyl cellulose hydroxyethyl cellulose and hydroxypropyl methyl cellulose METHOCEL; sucrose; dextrose; corn syrup; polysaccharides and gelatin. The binder may be selected from dry binders and / or wet granulation binders. Suitable dry binders are, for example, cellulose powder and microcrystalline cellulose, such as Avicel PH 102 and Avicel PH 200. In some embodiments, the oral composition comprises microcrystalline cellulose. Suitable binders for wet or dry granulation are corn starch, polyvinylpyrrolidone, vinylpyrrolidone-vinyl acetate copolymers and cellulose derivatives such as hydroxymethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose and hydroxypropylmethylcellulose. In some embodiments, the composition comprises povidone.

[0056] In some embodiments, the oral composition comprises 5-40% (w / w), such as 10-30% (w / w) or 5-25% (w / w) of filler. In some embodiments, the composition comprises 10.9% (w / w) or 18% (w / w) of filler, or comprises 19.5% (w / w) or 20.5 (w / w) of filler. The filler can be selected from lactose, mannitol, erythritol, sucrose, sorbitol, calcium phosphate such as calcium hydrogen phosphate, microcrystalline cellulose, powdered cellulose, powdered sugar, compressible sugar, dextrates, dextrins and dextrose. In some embodiments, the composition comprises microcrystalline cellulose, such as Avicel PH 102 or Avicel PH 200.

[0057] In some embodiments, the oral composition comprises 0.1-10% (w / w) or 0.5-5% (w / w), such as 1-3.5% (w / w) or 1% (w / w) of a lubricant and / or a glidant. In some embodiments, the composition comprises a lubricant and / or a glidant, such as talc, magnesium stearate, calcium stearate, zinc stearate, glyceryl behenate, polyethylene oxide polymer, sodium lauryl sulfate, magnesium lauryl sulfate, sodium oleate, sodium octadecyl fumarate, stearic acid, hydrogenated vegetable oil, silicon dioxide and / or polyethylene glycol. In some embodiments, the composition comprises magnesium stearate.

[0058] In some embodiments, the oral composition comprises a disintegrant, such as sodium starch glycolate, polacrilin potassium, sodium starch glycolate, crospovidone, cross-linked carboxymethylcellulose, sodium carboxymethylcellulose or dry corn starch. The composition may comprise one or more surfactants, such as one surfactant, at least one surfactant, or two different surfactants. The term "surfactant" refers to any molecule or ion consisting of a water-soluble (hydrophilic) portion and a fat-soluble (lipophilic) portion. The surfactant may, for example, be selected from anionic surfactants, cationic surfactants, nonionic surfactants and / or zwitterionic surfactants.

[0059] In some embodiments, the oral composition can be granulated before being compressed into tablets. In some embodiments, the granules of the present invention are manufactured by dry granulation, for example by roller compaction. In some embodiments, the molded product from the roller compaction process is crushed into granules. The composition may comprise one or more intragranular parts and extragranular parts, wherein the intragranular part has been granulated, and wherein the extragranular part has been added after granulation. The first intragranular part may comprise semaglutide and one or more excipients, and the second intragranular part may comprise a delivery agent and optionally one or more excipients. The first intragranular part may comprise semaglutide, a filler and / or a binder and the second intragranular part may comprise a delivery agent, a lubricant and / or a filler. In some embodiments, the first intragranular part comprises semaglutide, microcrystalline cellulose and / or povidone and the second intragranular part comprises a delivery agent, magnesium stearate and / or microcrystalline cellulose. The extragranular part may comprise a lubricant. In some embodiments, the extragranular part comprises magnesium stearate. In some embodiments, the filler and / or binder is a filler or a filler and a binder.

[0060] In some embodiments, to prepare a dry blend of tableting material, the various components are weighed, optionally de-lumped, and then combined. Mixing of the components may be performed until a uniform blend is obtained. If granules are used in the tableting material, the granules may be produced in a manner known to those skilled in the art, such as by dry granulation techniques, in which the pharmaceutically active agent and / or delivery agent is compressed together with the excipient to form a relatively large molding, such as a block or ribbon, which is comminuted by grinding, and the grounds are used as a tableting material to be later pressed into tablets.

[0061] In some embodiments, in order to compress the tablet material into a solid oral dosage form, such as a tablet, a tablet press can be used. In tableting, the tablet material is filled into a die cavity. The tablet material is then compressed by a punch with pressure. Subsequently, the resulting compact or tablet is ejected from the tablet press. Suitable tablet presses include, but are not limited to, rotary tablet presses and eccentric tablet presses.

[0062] In some embodiments, the method for preparing a tablet comprises: i) dry granulation of a mixture comprising semaglutide, a filler and a binder; ii) dry granulation of a mixture comprising a delivery agent, a lubricant and / or a filler; iii) mixing the granules with a lubricant, and then iv) compressing the blend into tablets.

[0063] In some embodiments, a method for producing an oral pharmaceutical composition comprises granules comprising a salt of N-(8-(2-hydroxybenzoyl)amino)caprylic acid and a lubricant (such as magnesium stearate), wherein the method comprises the steps of: a) mixing the salt of N-(8-(2-hydroxybenzoyl)amino)caprylic acid and the lubricant for more than 5 minutes; and b) optionally roller compacting the mixture of step a. In some embodiments, the present invention relates to a method for producing a pharmaceutical composition comprising a salt of N-(8-(2-hydroxybenzoyl)amino)caprylic acid and a lubricant (such as magnesium stearate), wherein the method comprises the steps of: a) mixing the salt of N-(8-(2-hydroxybenzoyl)amino)caprylic acid and the lubricant for more than 5 minutes; and b) optionally roller compacting the mixture of step a. In some embodiments, the mixing in step a is at least 6 minutes, for example at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 35, 40, 45 or 50 minutes. In some embodiments, the mixing in step a is at least 10 minutes, or at least 20 minutes. In some embodiments, the mixing in step a is at least 30 minutes or at least 40 minutes, for example at least 50 minutes. In some embodiments, the mixing in step a is in the scope of 6 minutes-12 hours, for example 10 minutes-10 hours or 15 minutes-8 hours. In some embodiments, the duration of the mixing described in step a is no more than 12 hours, for example no more than 10, 8, 6, 4 or 2 hours. In some embodiments, the amount of lubricant is at least 0.25% (w / w) or at least 0.5% (w / w), such as at least 1% (w / w) or at least 2% (w / w) of the composition. In some embodiments, the amount of lubricant is no more than 10% (w / w), such as no more than 8, 7, or 6% (w / w) of the composition.

[0064] In some embodiments, a method for preparing an oral pharmaceutical composition consisting of semaglutide and pharmaceutically acceptable excipients having the function of improving or alleviating dopamine dependence: 1) first sieving each ingredient through a #35 sieve; 2) geometrically mixing semaglutide and SNAC with a mortar and pestle; 3) dissolving povidone in water, and granulating the mixture of semaglutide and SNAC with the resulting solution; 4) drying the granules at a temperature not exceeding 40°C to a moisture content of ≤4%; and 5) grinding the obtained dried granules through a #35 sieve; 6) finally, mixing the granules with extragranular ingredients, and compressing the final mixture into tablets, wherein the compression is performed under a pressure of about 4.4 kN or higher.

[0065] In some embodiments, a pharmaceutical composition for injection comprising semaglutide is used to improve or alleviate dopamine dependence and inhibit the negative state produced by addiction, characterized in that the dopamine dependence includes: eating addiction, drinking addiction, coffee addiction, drug addiction and other behaviors closely related to dopamine.

[0066] In some embodiments, an oral pharmaceutical composition comprising semaglutide is used to improve or alleviate dopamine dependence and inhibit the negative state produced by addiction, characterized in that the dopamine dependence includes: eating addiction, drinking addiction, coffee addiction, drug addiction and other behaviors closely related to dopamine. BRIEF DESCRIPTION OF THE DRAWINGS

[0067] Figure 1 These are the results of the alcohol preference test for the control group (Control) and the model group (Model) eight weeks after modeling.

[0068] Figure 2 Blood alcohol levels in the control group (Control) and the model group (Model) eight weeks after modeling.

[0069] Figure 3 It is the percentage of the number of times the mice in the control group (Control) and the model group (Model) entered the open arm of the elevated plus maze test after the induction.

[0070] Figure 4 It is the percentage of time that mice in the control group (Control) and the model group (Model) spend in the open arm of the elevated plus maze test after induction.

[0071] Figure 5 This is the percentage of the number of times that mice in the treatment group (Treatmen), control group (Control) and model group (Model) entered the open arm of the elevated plus maze test after one week of semaglutide treatment.

[0072] Figure 6 This is the percentage of time that mice in the treatment group (Treatmen), control group (Control) and model group (Model) spent in the open arms of the elevated plus maze test after one week of semaglutide treatment.

[0073] Figure 7 The dopamine measurement results in the ventral tegmental area (VTA), control group (Control) n=3; model group (Model) n=5; alcohol withdrawal group (Withdrawal) n=5.

[0074] Figure 8 Results of the test for corticotropin-releasing factor (CRF) in the hypothalamus (n=3).

[0075] Fig. 9 The changes of blood corticosterone (CORT) during the drug administration period (n=5).

[0076] Fig.10 These are the test results of cyclic adenosine monophosphate (cAMP) in the hypothalamus, control group (Control) n=3; model group (Model) n=3; semaglutide treatment group (Treatmen) n=3. DETAILED DESCRIPTION

[0077] The present invention discloses a GLP-1 receptor agonist and a pharmaceutical composition thereof for improving or alleviating dopamine dependence. The following contents are embodiments of the specific technical solutions and do not limit the protection scope of the technical solutions. Example 1

[0078] A prescription composition and preparation method of an injectable pharmaceutical composition consisting of a GLP-1 receptor agonist and pharmaceutically acceptable excipients for improving or alleviating dopamine dependence.

[0079]

[0080]

[0081]

[0082] The general method for preparing the above formulation comprises: a) dissolving a preservative, an isotonic agent (propylene glycol) and a buffer in water and adjusting the pH to the desired pH; b) dissolving semaglutide in water while slowly stirring; c) mixing the two solutions and adjusting the pH to the desired pH using sodium hydroxide and / or hydrochloric acid; d) filtering the formulation through a 0.22 μm filter. Example 2

[0083] A prescription composition and a preparation method of an oral pharmaceutical composition consisting of a GLP-1 receptor agonist and pharmaceutically acceptable excipients for improving or alleviating dopamine dependence.

[0084]

[0085]

[0086]

[0087] Preparation of Tablet Formulation 1: Magnesium stearate for the first granulation fraction was passed through a 355 µm sieve. Magnesium stearate was manually mixed with SNAC in a stainless steel bowl of corresponding volume. Two cycles of geometric dilution were applied by mixing for about 60 s until the mixture was visually homogeneous. The remaining amount of SNAC was transferred to a mixer and premixed at 25 rpm for 2 min. The SNAC and magnesium stearate premix was added to the mixer and mixed at 25 rpm for 20 min. The mixture was roller compacted. The granules were sieved through a 180 µm sieve.

[0088] Semaglutide, microcrystalline cellulose and povidone for the second granulation fraction were weighed directly into a stainless steel bowl in decreasing order of amount and manually mixed for at least 3 min until visually homogeneous, and then the premix was transferred to a 1000 mL Duma bottle. The Duma bottle was closed with a lid and manually inverted in a Turbula-like motion for 1 min. The mixture was roller compacted.

[0089] The two types of granules were added to a mixing vessel in the order of decreasing content and mixed at 32 rpm for 5 minutes. Extragranular magnesium stearate was mixed with the granule mixture by manual mixing using volume-doubling, followed by mixing in a Turbula mixer at 32 rpm for 30 s. Tablets were prepared from this composition.

[0090] Preparation of Tablet Formulation 2: Magnesium stearate for granule fraction was passed through a 355 µm sieve. Magnesium stearate was manually mixed with SNAC in a stainless steel bowl of corresponding volume. Two cycles of geometric dilution were applied by mixing for about 60 s until the mixture was visually homogeneous. The remaining amount of SNAC was transferred to the mixer and premixed at 25 rpm for 2 min. The SNAC and magnesium stearate premix was added to the mixer and mixed at 25 rpm for 20 min. Semaglutide was geometrically diluted with the SNAC and magnesium stearate mixed in the mixer by mixing it manually for at least 60 s until visually homogeneous. The premix was then added to the mixed SNAC and magnesium stearate. The mixing step was completed by mixing in the mixer at 25 rpm for 10 min. The mixture was roller compacted.

[0091] The granules and all other components except extragranular magnesium stearate were added to a mixing vessel in the order of decreasing content and mixed at 32 rpm for 5 minutes. Extragranular magnesium stearate was mixed with the granule mixture by manual mixing using volume multiplication, followed by mixing at 32 rpm for 30 s in a Turbula mixer. Tablets were prepared from this composition.

[0092] Preparation method of tablet formulation 3: Magnesium stearate for granule fraction was passed through a 355 µm sieve. Magnesium stearate was manually mixed with SNAC in a stainless steel bowl of corresponding volume. Two cycles of geometric dilution were applied by mixing for about 60 s until the mixture was visually homogeneous. The remaining amount of SNAC was transferred to the mixer and premixed at 25 rpm for 2 min. The SNAC and magnesium stearate premix was added to the mixer and mixed at 25 rpm for 20 min. Semaglutide, microcrystalline cellulose and povidone were weighed into a stainless steel bowl in decreasing order of amount and the components were manually mixed until visually homogeneous. The geometric dilution was then performed using the mixed SNAC and magnesium stearate in the mixer by manually mixing it for at least 60 s until visually homogeneous. The premix was then added to the mixed SNAC and magnesium stearate. The mixing step was completed by mixing in the mixer at 25 rpm for 10 min. The mixture was roller compacted.

[0093] Extragranular magnesium stearate was mixed with the granules by manual mixing using volume doubling, followed by mixing in a Turbula mixer at 32 rpm for 30 s. Tablets were prepared from this composition. Example 3

[0094] Semaglutide can promote insulin secretion and reduce glucagon secretion, and has been approved for clinical use in the treatment of type II diabetes. Studies have shown that it can cross the blood-brain barrier and regulate various behaviors of mice by acting on different brain regions, such as cognitive function, anxiety and depression. Therefore, the present invention will take the semaglutide injection pharmaceutical composition as an example and explain its experimental effect in an animal model of alcohol use disorder (dopamine dependence model).

[0095] 1) Establishment of mouse alcohol addiction model 60 mice were divided into 12 cages, 5 mice in each cage. They were randomly divided into groups, of which 3 cages were blank groups, marked as blank group 1, blank group 2, and blank group 3. The other 9 cages were alcohol addiction induction groups, numbered 1-9. The mice in cages 1-9 were given different concentrations of alcohol solutions in stages, and the alcohol concentrations were 3% v / v. 6% v / v, 10% v / v, 15% v / v, and 20% vv. Two bottles of 3% vv alcohol solution were given on Monday, two bottles of water were given on Tuesday, and 3% v / v alcohol solution was given again on Wednesday. The same water was given on Thursday, and the same alcohol was given on Friday. When giving water or wine to two bottles, the positions of the two water bottles should be exchanged every day to prevent the mice from forming a position preference. Alcohol was removed on Saturday morning and water was not given at the same time, so that the mice were abstinent from alcohol and water for one day. The alcohol preference test was conducted at 9 am on Sunday, and the test time was 1.5 hours. After the test, the alcohol and water were replaced with two bottles of water for the mice to drink. According to this intermittent drinking method, the weekly alcohol concentration was increased to 6% v / v, 10% v / , 15% v / v, and 20% vv. Each alcohol concentration induced drinking for one week. When the drinking concentration was 20% vv, this concentration was maintained for four weeks. The modeling time was a total of 8 weeks, and an alcohol preference test was performed once a week on Sunday. At the end of the 8th week, alcohol preference was tested for two consecutive days, that is, two alcohol preference tests were performed on Monday and Tuesday of the 9th week. At the end of the last test, blood was collected from the mouse eye sockets to determine the alcohol concentration in the blood.

[0096] B) Alcohol preference test Before the alcohol preference experiment, mice were deprived of water and alcohol for one day in advance. Red Star Erguotou alcohol solution was used to prepare 2L of alcohol solution of the corresponding concentration for inducing drinking this week. The prepared alcohol solution was evenly distributed into the mouse water bottle, and the total weight of the corresponding water bottle and alcohol solution of each cage of mice was recorded. Each cage of mice corresponded to 2 bottles of alcohol solution. After 1.5 hours of drinking, the wine bottle was removed and weighed.

[0097] III) Methods of measuring blood alcohol concentration The alcohol concentration in the blood of mice that completed the alcohol addiction preference test was measured as follows: A. After the mice were anesthetized with ether, a 0.3mm capillary was inserted into the mouse's eye socket to collect 200μL of blood and allowed to stand for 2h. B. After standing, the blood was centrifuged at 4℃ and 3000rpm for 10min, and the supernatant was taken. C. 1mL of double distilled water was prepared, and reagent 2 was placed in an ice box for standby use. The other reagents were balanced to room temperature for 20min. D. The working solution of reagent 2 was prepared: 180L of double distilled water was added to each reagent 2 to dissolve and mix well. Reagent 2 and reagent 3 were mixed at a volume ratio of 1:17 and placed on an ice box for standby use. E. The working solution of the reaction was prepared: Reagent 1, reagent 2, and reagent 4 were mixed at a volume ratio of 10:5:1 and kept away from light for standby use. F. According to the preliminary experiment, the sample was diluted 5 times with double distilled water. G. The required strips were taken out of the aluminum foil that was balanced to room temperature. H. 40μL of standards of different concentrations were added to the corresponding wells. I. Take 40μL of the sample to be tested and add it to the measurement well. J. Add 160μL to the standard well and the measurement well. K. Immediate detection: vibrate the plate for 3s and measure the initial OD value of each well at a wavelength of 450nm using an ELISA reader. L. Incubate in a 37℃ oven away from light for 10min. M. Use an ELISA reader to measure the OD value A2 of each well at a wavelength of 450nm and calculate the OD change value. N. Calculate the ethanol content in the sample.

[0098] Experimental results of the mouse alcohol preference test and blood alcohol concentration: During the induction of modeling, the alcohol preference of mice was tested weekly, and the alcohol concentration in the blood of mice was detected after the alcohol preference test in the eighth week of modeling. The experimental data show that there was almost no difference in the preference for alcohol between the control group and the model group in the first two weeks, and the preference was about 20%. As the modeling time prolonged, the preference for alcohol in the model group continued to increase. In the fifth week, the alcohol preference of mice in each cage generally decreased. This may be because the alcohol concentration increased to 20% v / v, and the mice were not adapted to the high concentration of alcohol, causing the preference to drop suddenly. With the extension of the induction time, the alcohol preference of the control group and the model group changed greatly. Among them, the drinking preference of the control group continued to decrease with the increase of alcohol concentration, and its alcohol preference was less than 10% at the end of induction, at about 5%. In the model group, except for the mice in cages 1 and 9, whose alcohol preference was 35%, the alcohol preference of the other cages of mice reached about 60%.

[0099] After the modeling was completed, each group of mice was tested for alcohol preference. Figure 1 As shown. After the 8th week of induction, all mice were tested for alcohol preference three times with an alcohol concentration of 20% v / v. The results showed that the model group mice had a preference for alcohol of about 60%. After the last alcohol preference test, blood was collected from the eye sockets of mice in each cage to detect the alcohol content in the blood of mice after drinking. The results are shown as follows Figure 2As shown in the figure, the alcohol content in the blood of the model group mice after drinking reached more than 80mg / 100mL, reaching the standard of high-dose alcohol intake. The alcohol preference and blood alcohol content of the model group mice were significantly different from those of the control group. Example 4

[0100] The elevated plus maze (EPM) test was performed.

[0101] 5 days before the experiment, the mice were stroked for 5 minutes every day. The specific operation was as follows: A. 3 hours before the test, the mice to be tested were placed in the behavioral test room to eliminate the stimulation of the new environment on the mice and avoid the stimulation of the new environment causing large fluctuations in the experimental results, which in turn reduced the credibility of the results. B. Open the animal behavior analysis software and click to set the elevated cross maze program. C. Place the mouse in the central area of ​​the cross with its head facing the direction of the open arm, and click the software to start the test. D. After each mouse test, spray the elevated device with 5% acetic acid aqueous solution and wipe it dry with absorbent paper. E. Export the data from the software and return the mice to their original breeding place for subsequent experimental research. F. Process data: open arm entry number n1, closed arm entry number n2, open arm stay time t1, closed arm stay time t2. Percentage of open arm entry times = n1 / (n1+n2); percentage of open arm stay time = t1 / (t1+t2).

[0102] Elevated plus maze (EPM) experimental results: After 5 days of acute alcohol withdrawal in mice, all mice were subjected to an elevated cross test to test their anxiety levels after alcohol withdrawal. Figure 3 , Figure 4 As shown. After 5 days of acute alcohol withdrawal, the percentage of time that the Model group mice entered the open arms was significantly lower than that of the Control group, and the number of times they entered the open arms was significantly reduced. According to the principle of elevated cross behavioral detection: the less time and number of times the mice entered the open arms, the lower their desire to explore and the higher their anxiety value. T-test statistical analysis was performed on the percentage of time and number of times they entered the open arms, and it was found that there were significant differences between the Model group and the Control group. Figure 3 , Figure 4 The results showed that the mice in the Model group showed higher anxiety than those in the Control group during the alcohol withdrawal period after modeling. Example 5

[0103] The elevated plus maze test was conducted using the semaglutide injection pharmaceutical composition of prescription A in Example 1.

[0104] The mice that were addicted to alcohol were randomly divided into Treatmen group and Model group, with n=14 in Treatmen group, n=15 in Model group and n=15 in Control group. In the Treatmen group, the semaglutide drug composition was intrathecally injected, and the Model group and Control group were given an equal volume of sterile saline. The Treatmen group was administered for one week, on the 1st, 3rd, 5th and 7th days. During the administration period, all mice were only given water without alcohol. After 7 days of administration, open field and elevated cross experiments were performed on the 8th day to detect whether semaglutide improves the withdrawal symptoms of alcohol-addicted mice. On the 9th day, all mice were tested for alcohol preference and blood alcohol concentration to observe whether semaglutide improves the alcohol preference of addicted mice.

[0105] Results of the elevated plus maze test using the semaglutide injection pharmaceutical composition: At the same time, the mice in the Treatmen group, Control group and Model group were subjected to the elevated plus maze test. Figure 5 , Figure 6 As shown, Figure 5 It represents the percentage of mice entering the open arm within 5 min in the elevated plus maze test after one week of semaglutide treatment. Figure 6 It indicates the percentage of time that mice enter the open arms within 5 minutes after one week of drug administration. The results show that the Treatmen group entered the open arms more times and the time they entered the open arms were higher than the Model group. The results were significantly different, and the time and number of times that these two groups entered the open arms were lower than those of the Control group. The number and time that mice entered the open arms in the elevated plus maze experiment reflect the anxiety of the mice. The shorter the number of times they entered the open arms and the shorter the time they stayed in the open arms, the more anxious they were. From the results in the figure, it can be concluded that semaglutide can significantly improve the anxiety of mice addicted to alcohol after withdrawal, and thus improve the alcohol preference of addicted mice.

[0106] It can be seen from Examples 2-5 that after acute alcohol withdrawal, one week of semaglutide treatment can significantly improve the withdrawal symptoms of alcohol-addicted mice and reduce the alcohol preference of addicted mice. Example 6

[0107] Alcohol addiction is related to dopamine. In alcohol-addicted mice, dopamine secretion increases, which in turn causes the body to become excited and makes the addicted animals feel happy. When alcohol-addicted mice are acutely withdrawn from alcohol, the dopamine system and CRF system in the brain are affected, dopamine secretion in the brain decreases, and the CRF system is activated, which in turn causes an increase in adrenocorticotropic hormone-releasing factor. CRF is a neurotransmitter in the hypothalamus-pituitary-adrenal (HPA axis). It directly acts on the adrenocorticotropic hormone (ACTH) secreting cells of the pituitary gland to promote the secretion of ACTH, while ACTH acts on the adrenal gland to secrete corticosterone (rodents secrete corticosterone, and primates secrete cortisol). Corticosterone (CORT) acts on the corticosterone receptors on the surface of amygdala neurons to cause the body to feel anxious, which is why addicted mice experience withdrawal anxiety after acute alcohol withdrawal.

[0108] Experiment 1: Measurement of dopamine in the ventral tegmental area (VTA) of the mouse brain.

[0109] The specific operation steps are as follows: (1) Take the ventral tegmental area of ​​the mouse brain, cut it off 4mm before and after the mouse brain, put it into a 2mL EP tube, add 500μL of physiological saline, and grind it with a tissue grinder for 3min. Put the homogenate in 4℃, centrifuge and take the supernatant for later use. (2) Prepare washing solution - dilute according to the proportion. 20× washing buffer: distilled water = 1:19. Prepare 100mL of washing solution. (3) Take out the 96-well plate, draw the sample well and standard well area on the plate, and add 50mL of standard of different concentrations of 0pg / mL, 7.5pg / mL, 15pg / mL, 30pg / mL, 60pg / mL, and 120pg / mL to the standard well respectively; do 3 technical replicates for each concentration. (4) Add 10 μL of sample solution to the sample well, then add 40 μL of diluent to dilute the sample, add 100 μL of detection antibody (HRP) to the standard well and sample well, and place in a 37°C incubator for 60 minutes. (5) Pour out the liquid in the plate and add 350 mL of prepared washing solution to each well. Let stand for 1 minute, discard the washing solution and pat the remaining liquid in the well dry on filter paper. Repeat this operation 5 times. (6) Measure 2 mL of substrate A and substrate B each, mix them in a 10 mL centrifuge tube wrapped in tin foil, mix well, prepare them before use, add 100 mL of mixed substrate A and substrate B to each well, and place in a 37°C incubator away from light for incubation for 15 minutes. (7) Add 50 μL of stop solution to each reaction well, and use an ELISA reader to test the OD value of each well at a wavelength of 450 nm. (8) Export the data to create a standard curve and calculate the sample concentration.

[0110] Experiment 2: Determination of corticotropin-releasing factor (CRF) in mice.

[0111] The specific operation steps are as follows: (1) Take the hypothalamus from the brains of 3 mice in each group of the control group, treat group, and model group. (2) Put the taken hypothalamus (1 mg tissue: 9 mL PBS, add PBS and protease inhibitors: PBS = 1:99) into a homogenizer and grind it thoroughly on ice. Take out the homogenate and centrifuge the homogenate at 4°C, 5000×g for 10 minutes, and take the supernatant for later use. (3) Prepare washing solution - dilute according to the proportion. 20× washing buffer: distilled water = 1:19. Prepare 100mL of washing solution. (4) Take out the 96-well plate, draw the sample wells, standard wells, and blank well areas on the plate, and add 12.5pg / mL, 25pg / mL, 50pg / mL, 100pg / mL, 200pg / mL, 400pg / mL. 50mL of standards of different concentrations are added to the standard well area. (5) Add 10 mL of sample solution to the sample well, then add 40 mL of diluent to dilute the sample, add 100 mL of detection antibody (HRP) to the standard well and sample well, and place in a 37°C incubator for 60 minutes. (6) Pour out the liquid in the plate and add 350 mL of prepared washing solution to each well. Let stand for 1 minute, discard the washing solution and pat the remaining liquid in the well dry on filter paper. Repeat this operation 5 times. (7) Prepare a mixture of substrate A and substrate B. Use a pipette to draw 2 mL of substrate A and substrate B each and add them to a 5 mL EP tube. Then draw 100 mL of the mixture and add it to the above wells. Incubate at 37°C in the dark for 15 minutes. (8) Add 50 mL of stop solution to each reaction well, and use an ELISA reader to test the OD value of each well at a wavelength of 450 nm. (9) Export the data and make a standard curve to calculate the sample concentration.

[0112] Experiment 3: Detection of corticosterone (CORT) in the blood of mice.

[0113] The specific operation steps are as follows: (1) Take 100 mL of blood from the eye sockets of the mice in the blank group on the 6th day, the day before drug administration, the second day, the fourth day, and the sixth day of drug administration. (2) For sample processing, take 100 mL of whole blood from the mouse eye sockets and place it at room temperature for two hours. Then centrifuge at 1000×g for 20 minutes, and take the supernatant for use. (3) Prepare washing solution, dilute 10 mL of concentrated washing solution with distilled water to prepare 200 mL of working concentration washing solution, 20× washing buffer: distilled water = 1:19. (4) Take out the 96-well plate, mark the sample wells, standard wells, and blank wells on the plate, and add 50 mL of different concentrations of standard wells. (5) Add 10 mL of sample solution to the sample well, then add 40 mL of diluent to dilute the sample, add 100 mL of detection antibody (HRP) to the standard well and sample well, and place it in a 37°C incubator for 60 minutes. (6) Pour out the liquid in the plate and add 350 mL of prepared washing solution to each well. Leave for 1 minute, discard the washing solution and pat the remaining liquid in the well on the filter paper. Repeat this operation 5 times. (7) Prepare a mixture of substrate A and substrate B. Use a pipette to draw 2 mL of substrate A and substrate B respectively and add them to a 5 mL EP tube. Then draw 100 mL of the mixture and add it to the above wells. Incubate at 37°C in the dark for 15 minutes. (8) Add 50 mL of the reaction stop solution to each reaction well and use an enzyme-labeled instrument to test the OD value of each well at a wavelength of 450 nm. (9) Prepare a standard curve to calculate the sample concentration.

[0114] Experiment 4: Measurement of cyclic adenosine monophosphate (cAMP) in mouse hypothalamus.

[0115] The specific operation steps are as follows: (1) Take the hypothalamus from the brains of 3 mice in each group of the control group, treat group, and model group. (2) Put the taken hypothalamus (1 mg tissue: 9 μL PBS, add PBS and protease inhibitors: PBS = 1:99) into a homogenizer and grind it thoroughly on ice. Take out the homogenate and centrifuge the homogenate at 4°C, 5000×g for 10 min, and take the supernatant for later use. (3) Prepare washing solution - dilute according to the proportion. 20× washing buffer: distilled water = 1:19. Prepare 100mL of washing solution. (4) Take out the 96-well plate, draw the sample wells, standard wells, and blank wells on the plate, and add 50mL of different concentrations of standard wells of 0.0375pmol / mL, 0.75pmol / mL, 1.5pmol / mL, 3pmol / mL, 6pmol / mL, and 12pmol / mL. Perform three technical replicates for each concentration. (5) Add 10 mL of sample solution to the sample well, then add 40 mL of diluent to dilute the sample, add 100 mL of detection antibody (HRP) to the standard well and sample well, and place in a 37°C incubator for 60 minutes. (6) Pour out the liquid in the plate and add 350 mL of prepared washing solution to each well. Let stand for 1 minute, discard the washing solution and pat the remaining liquid in the well dry on filter paper. Repeat this operation 5 times. (7) Take 2 mL each of substrate A and substrate B and add them to a 5 mL EP tube and mix in the dark. Add 50 mL of the mixed substrate solution to each reaction well and incubate the 96-well plate at 37°C in the dark for 15 minutes. (8) Add 50 mL of stop solution to each reaction well and use an ELISA reader to test the OD value of each well at a wavelength of 450 nm. (9) Prepare a standard curve to calculate the sample concentration.

[0116] Experimental results: Results of dopamine measurement in the ventral tegmental area (VTA) of the brain: The dopamine in the ventral tegmental area of ​​the brain of mice in the blank group, model group, and alcohol withdrawal group was detected. The results are as follows Figure 7 As shown in the figure: Withdrawal group mice represent mice that have undergone acute alcohol withdrawal for 5 days, Control represents mice that are fed with normal water, and Model group mice are mice that have resumed normal drinking after the withdrawal. Figure 7It can be seen that the amount of dopamine in the VTA of the mice in the addiction group is higher than that in the blank group and the alcohol withdrawal group. There is a significant difference between the addiction group and the blank group, but no significant difference with the withdrawal group. In addition, although the dopamine content in the VTA of the alcohol withdrawal group is higher than that of the blank group, there is no significant difference. This shows that drinking alcohol in alcohol-addicted mice will increase the dopamine content in the brain, while acute alcohol withdrawal will reduce the dopamine content in the mouse brain. However, after the decrease, the concentration of dopamine in the brain is still higher than that in the normal group. This shows that when addicted mice undergo acute alcohol withdrawal, the dopamine in their brains will decrease. It is speculated that this may be because the mice do not consume alcohol, the activity of the brain's reward system is reduced, resulting in a decrease in the body's sense of pleasure and thus anxiety.

[0117] Results of the determination of CRF in the hypothalamus, corticosterone (CORT) in the blood, and cAMP in the hypothalamus of mice: The CRF levels in the hypothalamus of mice in the Control group, Model group, and Treatmen group were detected. Blood was collected from the orbits of mice in the Treatmen group one day before administration and on days 2, 4, and 6 during administration, and on the 6th day of administration, and the blood corticosterone content was measured. Then, whether the treatment with semaglutide changed the blood corticosterone content of addicted mice was compared. The experimental results are as follows: Figure 8 As shown, the CRF content in the hypothalamus of the Model group was higher than that of the Control group, indicating that the CRF system in the hypothalamus of alcohol-addicted mice was active and the CRF secretion increased during alcohol withdrawal. The CRF content in the hypothalamus of the Treatmen group was lower than that of the Model group. According to statistical analysis, the results of the two groups were significantly different. The experimental results showed that the administration of semaglutide can reduce the secretion of CRF in the hypothalamus of addicted mice during acute alcohol withdrawal. Fig. 9 It showed that the level of corticosterone in the blood was constantly decreasing during the treatment with semaglutide, but it was always higher than the level of corticosterone in the blood of mice in the Control group, and there was a significant difference between the two. The data of the Treatmen group one day before administration showed that semaglutide reduced the level of corticosterone in the blood of alcohol-addicted mice during acute alcohol withdrawal. Fig.10 The results showed that the cAMP in the hypothalamus of the Model group mice was higher than that of the Control group during alcohol withdrawal. The cAMP in the hypothalamus of the mice decreased after semaglutide treatment. Statistical analysis showed significant differences between the Model group and the Treatmen group.

[0118] Summarize: Since corticotropin-releasing factor is released by the hypothalamus in the hypothalamic-pituitary-adrenal (HPA) axis and acts on the pituitary gland, causing the pituitary gland to release ACTH, and ultimately ACTH acts on the adrenal glands to release CORT, excessive secretion of CORT can cause anxiety in the body.

[0119] Therefore, the CRF system in the hypothalamus of alcohol-addicted mice is activated during acute alcohol withdrawal, CRF secretion increases, and the corticosterone content in the blood also increases compared with the blank group. The administration of semaglutide treatment can reduce the secretion of CRF in the hypothalamus and reduce the corticosterone content in the blood. Combining this result with the elevated plus maze experiment in Examples 4 and 5, it is proved that semaglutide has the ability to improve or alleviate the symptoms of alcohol addiction, and can effectively inhibit negative emotional states such as anxiety that occur during acute alcohol withdrawal.

[0120] Although some features of the present invention have been illustrated and described herein, many modifications, substitutions, changes and equivalents will now occur to those skilled in the art. Therefore, it should be understood that the appended claims are intended to include all these modifications and changes that fall within the true spirit of the present invention.

Claims

1. A GLP-1 receptor agonist for improving or alleviating dopamine dependence, characterized in that: The GLP-1 receptor agonist is the sole pharmaceutically active ingredient or the sole GLP-1 receptor agonist administered to the subject.

2. A GLP-1 receptor agonist for improving or alleviating dopamine dependence according to claim 1, characterized in that: The GLP-1 receptor agonist is semaglutide.

3. A pharmaceutical composition comprising a therapeutically effective amount of the GLP-1 receptor agonist for improving or alleviating dopamine dependence as claimed in claim 1 and pharmaceutically acceptable excipients.

4. A pharmaceutical composition according to claim 3 comprising a therapeutically effective amount of the GLP-1 receptor agonist for improving or alleviating dopamine dependence according to claim 1 and a pharmaceutically acceptable excipient, characterized in that: The GLP-1 receptor agonist is the sole pharmaceutically active ingredient or the sole GLP-1 receptor agonist administered to the subject.

5. A pharmaceutical composition according to claim 3 comprising a therapeutically effective amount of the GLP-1 receptor agonist for improving or alleviating dopamine dependence according to claim 1 and a pharmaceutically acceptable excipient, characterized in that: The GLP-1 receptor agonist is semaglutide.

6. A pharmaceutical composition according to claim 3 comprising a therapeutically effective amount of the GLP-1 receptor agonist for improving or alleviating dopamine dependence according to claim 1 and a pharmaceutically acceptable excipient, characterized in that: The pharmaceutical composition is administered subcutaneously in the form of a solution containing at least 90% water and having a pH in the range of 7.0-9.

0.

7. A pharmaceutical composition according to claim 3 comprising a therapeutically effective amount of the GLP-1 receptor agonist for improving or alleviating dopamine dependence according to claim 1 and a pharmaceutically acceptable excipient, characterized in that: The pharmaceutical composition is administered orally, for example in the form of a tablet comprising at least 30% (w / w) of a salt of N-(8-(2-hydroxybenzoyl)amino)caprylic acid, such as SNAC.

8. Use of the pharmaceutical composition according to claims 3 to 6 in improving or alleviating dopamine dependence and inhibiting the negative state caused by addiction, characterized in that: The dopamine dependence mentioned includes: eating addiction, drinking addiction, coffee addiction, drug addiction and other behaviors closely related to dopamine.

9. Use of the pharmaceutical composition according to claims 3-5 and 7 in improving or alleviating dopamine dependence and inhibiting the negative state caused by addiction, characterized in that: The dopamine dependence mentioned mainly includes: eating addiction, drinking addiction, coffee addiction, drug addiction and other behaviors closely related to dopamine.

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