Application of IL-18 in preparation of medicine for preventing or treating post-traumatic stress disorder

Abstract

The invention discloses an application of IL-18 (interleukin-18) in preparation of a medicine for preventing or treating post-traumatic stress disorder. The invention creatively discovers that the IL-18 has a good effect in treating post-traumatic stress disorder (PTSD) for the first time, experiments prove that the IL-18 can effectively shorten the stiff time of a PTSD model mouse and relieve PTSD-like symptoms, a theoretical basis is provided for research and development of medicines for preventing or treating PTSD, a new application of the IL-18 is developed, and the IL-18 has a good application prospect. The invention provides a brand new method for treating PTSD, and has a wide application prospect.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of biological medicine, and particularly relates to application of IL-18 in preparation of a drug for preventing or treating post-traumatic stress disorder. BACKGROUND

[0002] Post-Traumatic Stress Disorder (PTSD) is a mental disorder that occurs after experiencing or witnessing an extraordinary threatening or disastrous traumatic event, and is a common mental disorder after traumatic events, which can last for months or even years. The main symptoms of PTSD include repeated experience related to the event, avoidance, negative changes in cognition and emotion, and excessive vigilance or arousal, which can cause social, biological and psychological health consequences, including social isolation, chronic pain and inflammation, cardiometabolic disorders, and increased risk of chronic dementia, accelerated aging and premature death, and increased risk of other mental disorders. PTSD seriously damages the physical and mental health and social function of individuals, and its adverse effects and long course of disease have caused heavy burden on individuals, families and society.

[0003] At present, Cognitive Behavioral Therapy (CBT) and Selective Serotonin Reuptake Inhibitors (SSRIs) are the preferred treatment options for PTSD. However, CBT has certain limitations, and it requires professional treatment personnel, specific treatment environment and high treatment cost in the implementation process. In terms of drug treatment, although SSRIs can alleviate some core symptoms of patients, they may cause adverse reactions such as sexual dysfunction and withdrawal symptoms. These factors significantly reduce the treatment compliance of patients. Due to the limitations of the above treatment methods, there is an urgent need for a treatment method with obvious effect and less side effects. The existing technology has confirmed that the disorder of the neuroimmune system plays a core role in the pathogenesis of PTSD, and a “pro-inflammatory state” exists in PTSD patients. However, the existing technology fails to distinguish the dual role of inflammatory response in different stages of the disease, and fails to develop and utilize the “homeopathic treatment” strategy of the body's own protective mechanism. SUMMARY

[0004] In order to make up for the deficiencies of the prior art, the purpose of the present application is to provide application of IL-18 in preparation of a drug for preventing or treating post-traumatic stress disorder.

[0005] In order to achieve the above-mentioned purpose, the technical scheme adopted by the present application is as follows: The present application provides, in a first aspect, the use of IL-18 in the preparation of a medicament for preventing or treating post-traumatic stress disorder.

[0006] In the present application, the IL-18 is also known as interferon-gamma inducing factor, which is a protein encoded by IL18 gene in human. The IL18 gene is located on human chromosome 11, and its gene ID in NCBI is 3606. The IL-18 protein is a proinflammatory cytokine of the IL-1 family, and its precursor exists in the cytoplasm of various cells including macrophages and keratinocytes. The inactive IL-18 precursor is converted into an active form after being processed by caspase-1, which can stimulate the production of interferon gamma and regulate helper T cell Th1 and Th2 responses. IL-18 is related to various organ injuries and potentially fatal diseases characterized by cytokine storm.

[0007] In some embodiments, the prevention refers to various means or measures for preventing the occurrence or development of diseases before being recognized by clinical standards, including medical, physical or chemical methods, to prevent and reduce the occurrence or development of various symptoms of diseases. In specific embodiments of the present application, the disease is preferably post-traumatic stress disorder.

[0008] In some embodiments, the treatment refers to the inhibition, suppression, alleviation, improvement, slowing, stopping, delaying or reversing of the development or aggravation of the disease, so that various indicators of the disease, disorder or pathological state described during the maintenance and / or administration are alleviated or reduced, including the alleviation or reduction of symptoms or complications, or the cure or elimination of the disease, disorder or condition. In specific embodiments of the present application, the disease is preferably post-traumatic stress disorder.

[0009] In specific embodiments of the present application, we have confirmed through animal experiments that exogenous supplementation of IL-18 can effectively alleviate the PTSD-like symptoms of PTSD mice induced by single prolonged stress (SPS) single prolonged stress, and the PTSD-like symptoms of mice are aggravated after administration of IL-18 binding protein or knock-out of IL-18R, further proving that the regulation of IL-18 can affect the PTSD-like symptoms of mice.

[0010] The present application provides, in a second aspect, the use of an IL-18 promoter in the preparation of a medicament for preventing or treating post-traumatic stress disorder.

[0011] In some embodiments, the IL-18 promoter of the present application refers to any substance capable of up-regulating the expression of IL-18, increasing the activity of IL-18, improving the stability of IL-18, increasing the effective action time of IL-18, or promoting the transcription and translation of IL-18, which falls within the scope of the present application.

[0012] In some embodiments, the substance capable of up-regulating the expression of IL-18 refers to any substance capable of up-regulating the expression of IL-18, including but not limited to: a naturally purified substance capable of up-regulating the expression of IL-18, a modified naturally purified substance, a semi-synthetic substance, a chemically synthesized substance, or any combination thereof.

[0013] Further, the IL-18 promoter includes an IL-18 overexpression plasmid vector or viral vector that promotes the expression of IL-18, an active peptide that promotes the expression of IL-18, an oligonucleotide that promotes the expression of IL-18, a protein that promotes the expression of IL-18, a small molecule compound that promotes the expression of IL-18, a nanoparticle carrying IL-18, or a liposome encapsulating IL-18.

[0014] The third aspect of the present application provides a medicament for preventing or treating post-traumatic stress disorder, the medicament comprising IL-18 and / or the IL-18 promoter of the second aspect of the present application.

[0015] In some embodiments, the medicament of the present application can be administered to a suitable subject once or more times per day. Unit dose means a physically discrete unit suitable for unit administration to a patient, and each unit contains a suitable pharmaceutical carrier and a predetermined quantity of IL-18 to provide a therapeutic effect. The dose varies depending on the severity of the patient's disease and the auxiliary effective ingredients used together. In addition, the total daily dose can be divided into several times and administered continuously as needed.

[0016] The fourth aspect of the present application provides a pharmaceutical composition for preventing or treating post-traumatic stress disorder, the pharmaceutical composition comprising the medicament of the first aspect of the present application as a first active ingredient.

[0017] Further, the pharmaceutical composition further comprises a second active ingredient capable of preventing, assisting in preventing, treating, or assisting in treating post-traumatic stress disorder.

[0018] Further, the second active ingredient includes an antidepressant, an anxiolytic drug, an antipsychotic drug, an anticonvulsant drug.

[0019] In some embodiments, the antidepressant drug is preferably selected from selective serotonin reuptake inhibitors (SSRIs), including sertraline, paroxetine, fluoxetine, escitalopram, fluvoxamine, etc.; the antidepressant drug is also selected from serotonin and norepinephrine dual reuptake inhibitors (SNRIs), including venlafaxine, duloxetine, etc., mirtazapine, tricyclic antidepressants, including amitriptyline, clomipramine, etc.; the anxiolytic drug is selected from benzodiazepines, including lorazepam, alprazolam, diazepam, clonazepam, etc.; the antipsychotic drug is selected from second-generation antipsychotics, including risperidone, olanzapine, quetiapine, amisulpride, etc.; the anticonvulsant drug is selected from sodium valproate, carbamazepine, etc.

[0020] In some embodiments, the second active ingredient which can be used for preventing, assisting in preventing, treating, assisting in treating post-traumatic stress disorder is not limited to the specific drugs listed above in the present application, any drug which can be used for preventing, assisting in preventing, treating, assisting in treating post-traumatic stress disorder will fall within the scope of the present application.

[0021] In some embodiments, the first active ingredient and the second active ingredient in the pharmaceutical composition can be administered simultaneously, separately or sequentially. Simultaneously means that the two drugs are administered at the same time. If not simultaneously, the administration is sequential within a time frame such that both are therapeutically active at the same time. Thus, sequential administration can allow administration of one drug 5 minutes, 10 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours or several hours after administration of the other, provided that the circulating half-life of the first administered drug is such that both are present in therapeutically effective amounts at the same time. The time delay between administration of the individual components will vary depending on the exact nature of the components, the interaction between them, and their respective half-lives. Separately means that there is a significant interval between administration of one drug and the other, i.e. when the second drug is administered, the first administered drug can no longer be present in the bloodstream in a therapeutically effective amount.

[0022] In some embodiments, the pharmaceutical composition is administered in a therapeutically effective amount in a manner compatible with the dosage formulation. The amount and time of administration depend on the subject to be treated, the ability of the subject to utilize the active ingredient, or the degree of therapeutic effect desired. The precise amount of active ingredient, such as the medicament of the third aspect of the present application or the pharmaceutical composition of the fourth aspect of the present application, required to be administered depends on the judgment of the physician, and is variable and will vary with the individual. Suitable dosing schedules are variable, but are represented by an initial administration, followed by subsequent injections or other administrations at intervals of one or more hours, or, alternatively, by continuous intravenous infusion sufficient to maintain the concentration in the blood within a range specified for in vivo therapy.

[0023] In some embodiments, the therapeutically effective amount refers to the amount of an active compound or pharmaceutical agent that elicits a biological or medical response sought by researchers, veterinarians, physicians, or other clinicians in a tissue, system, animal, or human. The therapeutically or pharmaceutically effective amount of the compound to be administered will be determined by such considerations and is the minimum amount necessary to improve, cure, or treat a disease or condition or one or more symptoms thereof. The pharmaceutical composition of the invention will be formulated, administered, and applied in a manner consistent with good medical practice, i.e., the dosage, concentration, regimen, process, medium, and route of administration. Factors considered in this context include the specific condition being treated, the specific mammal being treated, the individual patient's clinical condition, the cause of the condition, the site of delivery, the method of administration, the administration regimen, and other factors known to a medical practitioner, such as the individual patient's age, weight, and response.

[0024] The fifth aspect of the present invention provides a pharmaceutical preparation for the prevention or treatment of post-traumatic stress disorder, the pharmaceutical preparation comprising the medicament described in the third aspect of the present invention or the pharmaceutical composition described in the fourth aspect of the present invention.

[0025] Furthermore, the pharmaceutical preparation also contains pharmaceutically acceptable excipients.

[0026] In some implementations, "pharmaceutically acceptable" is used herein to mean a compound, material, composition, or dosage form that, to the extent reasonably medically judged, is suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications, and that meets a reasonable benefit / risk ratio.

[0027] In some embodiments, the excipients include binders, surfactants, flavoring agents, osmotic pressure regulators, antimicrobial agents, solubilizers, antioxidants, and stabilizers.

[0028] In some embodiments, the binder comprises carboxymethyl cellulose, methyl cellulose, hydroxypropyl methyl cellulose, polyethylene glycol, povidone, glycerin, polyvinyl alcohol, polyvinylpyrrolidone, polyalkyl styrene, polymethacrylate, and / or polyacrylate. The surfactant comprises hydroxypropyl-β-cyclodextrin, polyoxyethylene alkyl ether, Tween, sodium lauryl sulfate, sodium lauryl sulfate, poloxamer, polysorbate, sorbitan monopalmitate, sorbitan monostearate, sorbitan monooleate, and / or polyethylene glycol alkyl. The flavoring agent comprises sodium saccharin, cyclamate, aspartame, acetylsupan-K, glycerin, sorbitol, mannitol, sucrose, simple syrup, and aromatic syrup. The osmotic pressure regulator comprises sodium chloride, potassium chloride, mannitol, glycerin, and / or other pharmaceutically acceptable osmotic pressure regulators. The antibacterial agents include BAK, benzalkonium chloride, sorbic acid, oxychloride complex, citric acid, chlorobutanol, thimerosal, phenylmercuric acetate, disodium ethylenediaminetetraacetate, phenylmercuric nitrate, perborate, and / or benzyl alcohol. The cosolvents include hydrochloric acid, phosphoric acid, propionic acid, acetic acid, lactic acid, citric acid, tartaric acid, boric acid, glucuronic acid, gluconic acid, lactobionic acid, malic acid, threonic acid, glucoheponic acid, 2,5-dihydroxybenzoic acid, and acidic amino acids. The antioxidants and stabilizers include sulfurous acid, sulfites, bisulfites, metabisulfites, dithionite, thiosulfates, thiourea, glutathione, dimercaprol, mercaptoacetic acid and its salts, thiolactic acid and its salts, thiodipropionic acid and its salts, gallic acid and its salts, caffeic acid or its pharmaceutical salts, ferulic acid or its pharmaceutical salts, di-tert-butyl-p-phenol, 2,5-dihydroxybenzoic acid or its salts, salicylic acid or its salts, ascorbic acid and its salts, isoascorbic acid and its salts, nicotinamide, tartaric acid, phosphates, pharmaceutical salts of acetate, citrates, EDTA and its salts.

[0029] In some embodiments, the pharmaceutical formulation of the present invention may further include a drug delivery system, specifically including biodegradable polymers, biodegradable grafts, non-biodegradable grafts, biodegradable microparticles such as biodegradable microspheres, nanoparticles, etc.

[0030] In specific implementation schemes, the choice of which pharmaceutical excipients to combine with the drug formulation depends on the formulation to be made. When making ordinary formulations, the choice of which pharmaceutical excipients to use is well known to those skilled in the art.

[0031] Furthermore, the dosage form of the pharmaceutical preparation includes oral dosage form, parenteral dosage form, or topical dosage form.

[0032] In some implementations, oral administration refers to the absorption of a drug through the gastrointestinal tract after oral administration, thereby exerting its therapeutic effect. Dosage forms for oral administration include, but are not limited to: tablets, capsules, granules, powders, pills, syrups, oral solutions, suspensions, and emulsions. Parenteral administration refers to the delivery of a drug into the body via routes other than oral administration. Dosage forms for parenteral administration include, but are not limited to: injections, aerosols, powder sprays, sachets, patches, and implants. Local administration refers to the application of a drug to local tissues to achieve a local therapeutic effect. Dosage forms for local administration include, but are not limited to: topical solutions, lotions, liniments, ointments, and gels.

[0033] In some embodiments, the present invention does not impose any particular limitation on the dosage form of the drug, and the drugs in all the above dosage forms can be prepared according to conventional methods in the pharmaceutical field.

[0034] In some embodiments, the drug, pharmaceutical composition, or pharmaceutical preparation described herein may be administered to the subject via injection, topical administration, or oral administration. For example, the method may include administering the drug, pharmaceutical composition, or pharmaceutical preparation to the subject three times a day, once a day, or every two days. In some embodiments, injection administration may include subcutaneous injection, intramuscular injection, intravenous injection, etc. In some embodiments, injection administration may include directly injecting the drug, pharmaceutical composition, or pharmaceutical preparation into or near the lesion. In some embodiments, topical administration may include rectal administration, nasal administration, ear administration, intramedullary administration, intra-articular administration, intrapleural administration, etc., or any combination thereof. In some embodiments, the drug, pharmaceutical composition, or pharmaceutical preparation may be administered to the subject via a combination of different administration methods.

[0035] In some implementations, the patient / subject can be human or non-human and can include, for example, animal strains or species used as a "model system" for research purposes. Similarly, the patient / subject can include adults or adolescents (e.g., children). Furthermore, the patient / subject can refer to any living organism, preferably a mammal (e.g., human or non-human). Examples of mammals include, but are not limited to, any member of the mammalian class: humans, non-human primates (e.g., chimpanzees) and other apes and monkeys; livestock such as cattle, horses, sheep, goats, and pigs; domestic animals such as rabbits, dogs, and cats; and laboratory animals including rodents such as rats, mice, and guinea pigs. Examples of non-mammals include, but are not limited to, birds, fish, etc.

[0036] Advantages and beneficial effects of the present invention: This invention is the first to creatively discover that IL-18 has a good effect in the treatment of PTSD. Through experiments, this invention demonstrates that IL-18 can effectively shorten the stiffness time in PTSD model mice and alleviate PTSD-like symptoms. This invention provides a theoretical basis for the development of drugs for the prevention or treatment of PTSD, opens up new uses for IL-18, and provides a brand-new method for the treatment of PTSD, with broad application prospects. Attached Figure Description

[0037] Figure 1 The graph shows the evaluation of the SPS model and the expression level of IL-18. In the graph, A is the experimental protocol; B is the time of mouse stiffness on day 8 of modeling; C is the time of mouse stiffness on day 15 of modeling; and D is the IL-18 expression level of mice detected by ELISA.

[0038] Figure 2 The diagram shows the therapeutic effects of IL-18. In the diagram, A is the experimental protocol; B is the resting time of mice in each group treated with IL-18 on day 8 of modeling; C is the resting time of mice in each group treated with IL-18 on day 15 of modeling; D is the resting time of mice in each group treated with IL-18 binding protein on day 8 of modeling; and E is the resting time of mice in each group treated with IL-18 binding protein on day 15 of modeling.

[0039] Figure 3 The figures show the resting time of mice after IL-18R knockdown. In the figures, A is the experimental protocol diagram; B is the resting time of mice in each group with IL-18R knockdown on day 8 of modeling; and C is the resting time of mice in each group with IL-18R knockdown on day 15 of modeling. Detailed Implementation

[0040] As used in this invention, the terms “having,” “comprising,” or “including,” or any grammatical variations thereof, are used in a non-exclusive manner. Thus, these terms can all refer to a situation where no other features exist in the entity described in this context besides the features introduced by these terms, and can also refer to a situation where one or more other features are present.

[0041] Furthermore, as used in this invention, the terms “preferred,” “more preferred,” “most preferred,” “particularly,” “more particularly,” “specifically,” “more specifically,” or similar terms are used in combination with optional features without limiting other possibilities.

[0042] Unless otherwise stated, all figures used in this specification and claims to represent volume, weight, temperature, time, density, parts by weight, technical effect, etc., should in any case be understood to be modified by the terms "about" or "approximately". Therefore, unless indicated to the contrary, the numerical parameters listed in the specification and appended claims are approximate values. Unless otherwise stated, the terminology used herein has its common meaning as understood by one of ordinary skill in the art, and each numerical parameter should be interpreted according to the number of significant figures and conventional rounding methods or in a manner understood by one of those skilled in the art.

[0043] Example: Study on the efficacy of IL-18 in treating post-traumatic stress disorder I. Experimental Methods 1. Experimental materials (1) Experimental animals: 8-12 week old male C57BL / 6N wild-type mice, purchased from Vital River.

[0044] (2) Main equipment: Panlab SLU shock condition system, forced swimming equipment (Reward Life Science Co., Ltd.), anesthesia machine (Reward Life Science Co., Ltd.), bilateral drug delivery cannula (Reward Life Science Co., Ltd.), brain localization injection device (Reward Life Science Co., Ltd.).

[0045] (3) Main reagents and consumables: Mouse IL-18 ELISA kit (MBL), IL-18 (MCE), IL-18 binding protein (MCE), AAV virus (Wuhan Shumi Brain Science Technology Co., Ltd.): rAAV-GFAP-EGFP-5'miR-30a-shRNA1(I118r1)-3'-miR30a-WPRES, rAAV-Iba1-EGFP-5'miR-30a-shRNA1(I118r1)-3'miR-30a-WPRES-4XMIR-9T, rAAV-hSyn-EGFP-5'miR-30a-shRNA1(I118r1)-3'-miR30a-WPRES, AAV-EF1a-EGFP-5'miR-30a-shRNA(scramble)-3'-miR30a-WPREs.

[0046] 2. Experimental Procedure (1) Single Stress Model (SPS) Modeling: Mice were placed in the laboratory for acclimatization 1 hour before the experiment. Mice were restrained for 2 hours in 50 ml centrifuge tubes with ventilation holes. Animals were placed in a forced swimming tub with the ideal water temperature controlled at 23-25℃ (initial water temperature 25℃). Each animal was monitored until exhaustion, about 15 minutes, dried, and rested for 15 minutes. Animals were placed in a transparent anesthesia box and anesthetized with ether to control the depth of anesthesia. After confirming the disappearance of reflexes, they were removed and allowed to rest until awake. Animals were placed in a conditioned fear system. The experimental group program was set as follows: spontaneous activity for 5 minutes, electric shock for 10 seconds, once, current 0.8 mA, spontaneous activity for 5 minutes. The control group program was set as follows: spontaneous activity for 10 minutes. On the 8th and 15th days after modeling, animals were placed in the laboratory for 2 hours for acclimatization and then placed back in the conditioned fear box for 5 minutes of spontaneous activity. Animal behavior was mainly detected by gravity sensors, and the duration of the mouse's rigid state was recorded.

[0047] (2) ELISA: The dorsal hippocampus tissue of mice was isolated on ice. The tissue isolated from each mouse was placed in 250 μl of tissue lysis buffer (protease inhibitor added at 1:100), and 2-5 grinding beads were added. The mixture was ground at 60 Hz for 60 s, then centrifuged at 5000 rpm at 4℃ for 5 minutes. The supernatant was collected and centrifuged at 15000 rpm at 4℃ for 5 minutes. The supernatant was collected and stored. IL-18 was detected using the mouse IL-18 ELISA kit according to the instructions.

[0048] (3) Installation of the drug delivery cannula: Anesthetize mice with isoflurane using an anesthesia device, and shave the hair on the head from the eyes to behind the ears. Cut open the skin of the skull, wipe the incision with a cotton swab dipped in physiological saline, and clean the skull with a cotton swab dipped in distilled water to fully expose the anterior and posterior fontanelles, ensuring that the skull is clean (free of blood, fur, or tissue) and dry (use a dry cotton swab to absorb any distilled water or blood), estimate and mark the position of the anterior fontanelle. Adjust the anterior, posterior, left, and right angles of the animal's head so that the anterior and posterior fontanelles are at the same level, and the left and right sides are at the same level. First, determine that the anterior and posterior fontanelles are consistent with the anterior-posterior direction of the brain localization instrument, then adjust the left and right sides to be level, with the difference in the left and right digital display values ​​not exceeding 0.03. Then determine that the anterior and posterior fontanelles are at the same level. Use the mouse brain stereotaxic atlas to determine the location of the experimental target area, and mark the point with the tip of the syringe needle. In this experiment, the coordinates of the hippocampal brain region are AP = -2.2 mm, ML = ±1.3 mm, and DV = -2 mm. Using a skull drill, a small hole slightly larger than the diameter of the cannula is drilled in the target area, taking care not to damage the dura mater. The bone fragment is carefully removed from the hole using the tip of a 1 mL syringe needle, exposing the cortex. The cannula holder is mounted on a stereotactic frame, and the drug delivery cannula is inserted into the target area according to the predetermined coordinates. It is then fixed with dental cement. After one week of postoperative recovery, model construction and testing begin.

[0049] (4) IL-18 and IL-18 binding protein administration: Pre-assemble the injection tubing, PE tubing, locking cap, and syringe. Use an infusion pump to draw up the drugs. In the IL-18 administration experiment, each mouse was injected with a total of 0.5 μg of IL-18 per injection, while the control group was injected with an equal volume of PBS. In the IL-18 binding protein administration experiment, each mouse was injected with a total of 0.1 μg of IL-18 binding protein per injection, while the control group was injected with an equal volume of PBS. Mark the drug level on the PE tubing and observe whether the drug level drops during the injection process. Remove the catheter cap, slowly insert the injection tubing into the catheter, and secure it with the locking cap. Set the injection volume and rate of the infusion pump: total volume per side per injection: 250 nl, injection rate: 0.15 μl / min. After the injection is completed, wait 10 minutes to allow the drug to fully diffuse, then slowly pull out the injection tubing, reinsert the catheter cap, and tighten it. After SPS modeling, IL-18 was administered from the day of modeling until the end of testing on day 15. IL-18 binding protein was administered three days before the modeling day until the end of testing on day 15 after modeling, with one dose given bilaterally daily.

[0050] (5) IL-18R knockdown experiment: Place a hard glass tube in a glass microelectrode drawing instrument and adjust the parameters to draw the glass tube into an electrode with a tip length of about 5 mm for virus injection in one step. Soak the drawn glass electrode in 75% alcohol for 30 min, then soak it in double-distilled water for 10 min. Dry it in a clean bench and store it in a special glass electrode storage box. Install the electrode on a microinjection pump and remove air bubbles from the electrode tip. Thaw the required virus on ice and draw the virus using a microinjection pump. Anesthetize the mice with isoflurane using an anesthesia device and shave the hair from the eyes to behind the ears. Cut open the skull skin and wipe the incision with a cotton swab dipped in physiological saline. Clean the skull with a cotton swab dipped in distilled water to fully expose the anterior and posterior fontanelles, ensuring that the skull is clean (free of blood, fur or tissue) and dry (use a dry cotton swab to absorb any distilled water or blood). Estimate and mark the position of the anterior fontanelle. Adjust the front-back and left-right angles of the animal's head so that the anterior and posterior fontanelles are at the same level and the left and right sides are at the same level. First, ensure the anterior and posterior fontanelles are aligned with the anteroposterior direction of the brain localization instrument. Then, adjust the left and right sides to be level, ensuring the difference in digital display values ​​between the left and right sides is no greater than 0.03. Next, ensure the anterior and posterior fontanelles are at the same level. Use a mouse brain stereotaxic atlas to determine the location of the experimental target area and mark the points with a syringe needle tip. In this experiment, the coordinates of the hippocampus are AP = -2.2 mm, ML = ±1.3 mm, and DV = -2 mm. Carefully and slowly drill a small hole in the target area using a skull drill. Carefully remove the bone fragment from the hole with a 1 mL syringe needle tip to expose the cortex. Install the microinfusion pump on the stereotaxic instrument and insert the glass electrode containing the virus into the target area according to the predetermined coordinates. Inject 450 μL of virus per side at a rate of 0.15 μl / min, with a viral titer of 2.00 × 10⁻⁶. 12 Vg / ml, let stand for about 10 minutes, then slowly remove the electrode. After injection, suture the mouse scalp completely with a suture needle and place it in a rearing cage for recovery. Modeling and behavioral testing will begin 3 weeks after surgery.

[0051] II. Experimental Results The results showed that the freezing time of the model group mice was significantly longer than that of the control group on both day 8 and day 15, indicating that the mice exhibited obvious PTSD-like fear symptoms, proving that the model was successfully established. Figure 1 A, B, C). IL-18 ELISA results showed that IL-18 in the dorsal hippocampus of the model group mice was significantly upregulated at 3, 8, and 15 days. Figure 1 D).

[0052] To investigate whether IL-18 is involved in regulating PTSD-like fear symptoms, IL-18 or IL-18-binding protein was continuously administered to the dorsal hippocampus, and the effect on the maintenance of fear memory at 8 and 15 days after modeling was observed. The results showed that the freezing time in the IL-18 group was significantly shorter than that in the PBS control group at both days 8 and 15, indicating that PTSD-like fear symptoms were alleviated in the mice. Figure 2 Mice in groups A, B, and C treated with IL-18 binding protein showed significantly longer periods of stiffness on days 8 and 15 compared to the PBS control group, indicating that the mice exhibited enhanced fear symptoms compared to the control group. Figure 2 The results of the two drug administration experiments (A, D, and E) corroborate each other, indicating that the regulation of IL-18 can affect PTSD-like symptoms in mice.

[0053] PTSD-like fear symptoms in mice were significantly enhanced (days 8 and 15) after AAV virus-specific interference with neuronal IL-18 receptors, demonstrating that targeting the IL-18 / IL-18R pathway is an effective means of regulating PTSD. Figure 3 ).

[0054] The above description of the embodiments is only for understanding the method and core ideas of the present invention. It should be noted that those skilled in the art can make various improvements and modifications to the present invention without departing from the principles of the invention, and these improvements and modifications will also fall within the protection scope of the claims of the present invention.

Claims

1. The use of IL-18 in the preparation of drugs for the prevention or treatment of post-traumatic stress disorder.

2. Application of IL-18 promoters in the preparation of drugs for the prevention or treatment of post-traumatic stress disorder.

3. The application according to claim 3, characterized in that, The IL-18 promoters include IL-18 overexpression plasmid vectors or viral vectors that promote IL-18 expression, active peptides that promote IL-18 expression, oligonucleotides that promote IL-18 expression, proteins that promote IL-18 expression, small molecule compounds that promote IL-18 expression, nanoparticles carrying IL-18, or liposomes encapsulating IL-18.

4. A drug for the prevention or treatment of post-traumatic stress disorder, characterized in that, The drug includes IL-18 and / or the IL-18 promoter of claim 2 or 3.

5. A pharmaceutical composition for the prevention or treatment of post-traumatic stress disorder, characterized in that, The pharmaceutical composition comprises the drug of claim 4 as the first active ingredient.

6. The pharmaceutical composition according to claim 5, characterized in that, The pharmaceutical composition also contains other second active ingredients that can be used for prevention, adjunctive prevention, treatment, and adjunctive therapy of post-traumatic stress disorder.

7. The pharmaceutical composition according to claim 6, characterized in that, The second active ingredient includes antidepressants, anxiolytics, antipsychotics, and anticonvulsants.

8. A pharmaceutical preparation for the prevention or treatment of post-traumatic stress disorder, characterized in that, The pharmaceutical preparation comprises the drug of claim 4 or the pharmaceutical composition of any one of claims 5-7.

9. The pharmaceutical preparation according to claim 8, characterized in that, The pharmaceutical preparation also contains pharmaceutically acceptable excipients.

10. The pharmaceutical preparation according to claim 8, characterized in that, The dosage forms of the pharmaceutical preparations include oral dosage forms, parenteral dosage forms, or topical dosage forms.

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