Application of YL-0919 in treatment of epilepsy or brain injury caused by epilepsy

Abstract

The invention discloses an application of YL-0919 in treatment of epilepsy or brain injury caused by epilepsy. Animal experiments prove that YL-0919 has a good effect of treating epilepsy or brain injury caused by epilepsy. The research result of the invention provides a new method and thought for clinical treatment.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology. Specifically, the present invention relates to a new use of a known compound. More specifically, the present invention relates to the use of YL-0919 in the treatment of epilepsy or brain damage caused by epilepsy. Background Art

[0002] Epilepsy is a chronic brain disease characterized by recurrent seizures. It can occur suddenly and without cause. It is one of the most common chronic neurological diseases. The incidence rate in the Chinese population is between 5% and 7%. Its pathogenesis is complex and has a great impact on the lives of patients. Although there are a variety of anti-epileptic drugs that can be used for the symptomatic treatment of epilepsy, about one-third of patients suffer from drug-refractory epilepsy.

[0003] Hydroxypiperidone hydrochloride (YL-0919) is a new small molecule antidepressant that exerts significant antidepressant and antianxiety-like effects in various animal models. Studies have shown that YL-0919 is a new 5HT1A receptor partial agonist. However, there is still a gap in the research of YL-0919 in the treatment of epilepsy or brain damage caused by epilepsy. Summary of the invention

[0004] The purpose of the present invention is to propose a new use of YL-0919 in drugs for treating epilepsy or brain damage caused by epilepsy, so as to make up for the shortcomings of the prior art.

[0005] The above-mentioned object of the present invention is achieved through the following technical solutions: In a first aspect, the present invention provides a drug for treating epilepsy or brain damage caused by epilepsy.

[0006] Furthermore, the drug comprises an effective dose of 5HT1A receptor agonist YL-0919 or a pharmaceutically acceptable salt thereof.

[0007] In the present invention, the 5HT1A receptor partial agonist includes but is not limited to Buspirone, Tandospirone, Ipsapirone, Aripiprazole, and YL-0919.

[0008] Furthermore, the 5HT 1A The receptor agonist is YL-0919.

[0009] Furthermore, the drug can be prepared in the form of tablets, capsules, granules, pills, pellets, syrups, powders, granules, suppositories, drops, emulsions, injections, solutions or suspensions.

[0010] In the present invention, the "YL-0919" is a new structural compound in powder form with a molecular formula of C 18 H 22 N2O2·ClH, with a molecular weight of 334.84, is a 5HT1A receptor agonist and 5-HT uptake blocker, showing significant antidepressant effects in animal models. YL-0919 has been shown to exert rapid antidepressant and antianxiety effects, enhance cognitive behavior in mice, and improve ischemic stroke in rats. There are currently no studies or reports on the use of YL-0919 in the treatment of epilepsy or brain damage caused by epilepsy, and this is the first report.

[0011] In the present invention, the "effective dose" is an appropriate dose that is non-toxic to the patient and supported by data obtained from experimental studies, but it does not exclude that in some cases, a larger dose that may cause signs of toxicity may be given according to the severity of the disease. "Effective dose" refers to the amount of YL-0919 of the present invention: it is sufficient to prevent the development or onset of epilepsy or one or more symptoms thereof, enhance or improve the effect of another treatment, and / or alleviate one or more symptoms of epilepsy. For subjects suffering from epilepsy, the preferred therapeutically effective amount is an amount that effectively reduces the frequency of epileptic seizures and / or reduces pathological damage to hippocampal neurons.

[0012] In some embodiments, a therapeutically effective amount can be administered to a patient in one or more doses, the dosage being sufficient to alleviate, mitigate, stabilize, reverse or slow the progression of the disease, or otherwise mitigate the pathological consequences of the disease, or mitigate the symptoms of the disease. The mitigation or alleviation does not need to be lasting, but can be within a certain time frame (at least one hour, at least one day, or at least one week or longer). The effective amount is usually determined by a doctor according to the individual case, and is within the skill range of those skilled in the art. When determining the appropriate dose for achieving an effective amount, several factors are usually considered. These factors include the patient's age, sex and weight, the condition being treated, the severity of the condition, and the route of administration, dosage form and regimen, and the desired result.

[0013] In some embodiments, the dosage depends on many factors, such as the nature and severity of the disease to be prevented and / or treated (in the present invention, especially epilepsy or brain damage caused by epilepsy), the sex, age, weight and individual response of the patient or animal, the specific compound used, the route of administration and the number of administrations, etc. The above dosage can be administered in a single dosage form or divided into several, such as two, three or four dosage forms.

[0014] Furthermore, a variety of prescriptions can be made based on factors such as the formulation method, administration method, patient's age, weight, sex, morbidity, diet, administration time, administration route, excretion rate and reaction sensitivity, and a skilled physician can usually easily determine the prescription and the dosage that is effective for the desired prevention and / or treatment.

[0015] The term "pharmaceutically acceptable salt" refers to pharmaceutically acceptable salts of YL-0919, which means those carboxylates, amino acid addition salts, etc. of the compound (YL-0919) described in the present invention, which are suitable for contact with patients within the scope of reliable medical judgment, do not produce undue toxicity, irritation, allergic reactions, etc., are commensurate with a reasonable benefit / risk ratio, and are effective for their intended use, including (if possible) zwitterionic forms of the compound (YL-0919) described in the present invention.

[0016] In some embodiments, examples of the "pharmaceutically acceptable salt of YL-0919" include, but are not limited to, salts having (as counter ions) alkali metal ions such as Li + 、Na + or K + or salts with alkaline earth metal ions such as Mg 2+ or Ca 2+ or any other pharmaceutically acceptable metal ion such as Zn 2+ or Al 3+ or a pharmaceutically acceptable salt formed with an organic base such as diethanolamine, ethanolamine, N-methylglucamine, triethanolamine or tromethamine.

[0017] In some embodiments, pharmaceutically acceptable base addition salts are formed with metals or amines, such as alkali metals and alkaline earth metal hydroxides or organic amines. Examples of metals used as cations are sodium, potassium, magnesium, calcium, etc. Examples of suitable amines are N, N'-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, N-methylglucamine and procaine.

[0018] In some embodiments, base addition salts of acidic compounds can be prepared by contacting the free acid form with a sufficient amount of the desired base in a conventional manner to form the salt. The free acid can be regenerated by contacting the salt form with an acid in a conventional manner and then isolating the free acid. The free acid forms differ from their respective salt forms somewhat in certain physical properties, such as solubility in polar solvents, but for purposes of the present invention, the salts are equivalent to their respective free acids.

[0019] In some embodiments, the salt can be a sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide prepared from an inorganic acid, such as hydrochloric acid, nitric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, phosphoric acid, etc. Representative salts include: hydrobromide, hydrochloride, sulfate, bisulfate, nitrate, acetate, oxalate, valerate, oleate, palmitate, stearate, laurate, borate, benzoate, lactate, phosphate, toluenesulfonate, citrate, maleate, fumarate, succinate, tartrate, naphthoate, methanesulfonate, glucoheptonate, lactobionate, lauryl sulfonate, and isethionate, etc. Salts can also be prepared from organic acids, such as aliphatic mono- and dicarboxylic acids, phenyl-substituted alkanoic acids, hydroxyalkanoic acids, alkanedioic acids, aromatic acids, aliphatic and aromatic sulfonic acids, etc. Representative salts include acetate, propionate, octanoate, isobutyrate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, mandelate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, naphthoate, benzenesulfonate, toluenesulfonate, phenylacetate, citrate, lactate, maleate, tartrate, methanesulfonate, etc. Pharmaceutically acceptable salts may include cations based on alkali metals and alkaline earth metals, such as sodium, lithium, potassium, calcium, magnesium, etc., as well as non-toxic ammonium, quaternary ammonium and amine cations, including but not limited to ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine, etc. Also contemplated are salts of amino acids, such as arginate, gluconate, galacturonate, etc.

[0020] The administration of the drug YL-0919 of the present invention includes subcutaneous injection, intradermal injection, intramuscular injection, intraperitoneal injection, intravenous injection, oral administration, etc. In the past, oral administration was used in the study of beagle dogs, and intraperitoneal injection was used in the study of rat stroke. The present invention preferably uses intraperitoneal injection to improve the stability of the drug in mice.

[0021] In a second aspect, the present invention provides use of a 5HT1A receptor agonist or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating epilepsy or brain damage caused by epilepsy.

[0022] The 5HT1A receptor partial agonists of the present invention include but are not limited to Buspirone, Tandospirone, Ipsapirone, Aripiprazole, YL-0919, and the like.

[0023] Furthermore, the 5HT1A receptor agonist is YL-0919.

[0024] Furthermore, the effect of the drug is to reduce the frequency of epileptic seizures.

[0025] Furthermore, the effect of the drug is to reduce the degree of inflammation in the hippocampus.

[0026] Furthermore, the effect of the drug is to reduce pathological damage to hippocampal neurons.

[0027] Furthermore, the epilepsy includes primary epilepsy, secondary epilepsy, and cryptogenic epilepsy.

[0028] Preferably, the epilepsy is secondary epilepsy.

[0029] More preferably, the secondary epilepsy is temporal lobe epilepsy.

[0030] The causes of epilepsy described in the present invention include but are not limited to muscle contraction, brain damage, genetic factors, physical illness, drug abuse, etc. The present invention is epilepsy caused by brain damage, which is the most important cause of epilepsy.

[0031] The establishment of epilepsy or brain damage caused by epilepsy in the present invention requires the construction of a mouse model. The methods for constructing an epilepsy mouse model include, but are not limited to, a kindling model, a pentylenetetrazol (PTZ) epilepsy model, a kainic acid (KA) epilepsy model, a maximum electroshock (MES) model, a gene knockout, and a transgenic epilepsy animal model. The present invention preferably uses a kainate (KA) epilepsy model, because the KA model is similar to humans in terms of latency, behavioral symptoms, latency duration, and electroencephalogram characteristics during the latency and chronic phases, and is more valuable for research.

[0032] The method for judging epilepsy or brain damage caused by epilepsy in the present invention involves multiple aspects, including but not limited to observation of neurobehavioral characteristics, analysis of brain tissue morphological characteristics, spongiform changes, density of neuronal dendritic spines, expression of inflammation-related genes, detection of inflammatory protein expression, electroencephalogram analysis, cerebral blood flow analysis, degree of hippocampal inflammation, neuronal pathological damage, etc. The present invention analyzes the degree of inflammation and neuronal pathological damage in the hippocampus, which is an important indicator for judging brain damage in mice.

[0033] Secondary epilepsy caused by brain damage includes but is not limited to temporal lobe epilepsy, epilepsy caused by brain tumors, cerebral hemorrhage, cerebral infarction, viral encephalitis, autoimmune encephalitis, and localized cortical dysplasia. The present invention is the most common temporal lobe epilepsy among secondary epilepsy, and is also the most common type of brain damage caused by epilepsy or epilepsy.

[0034] Advantages and beneficial effects of the present invention: The present invention discovers for the first time that YL-0919 can be used to treat epilepsy or brain damage caused by epilepsy, providing a new perspective and method for the clinical treatment of epilepsy or brain damage caused by epilepsy.

[0035] The present invention uses a mouse model to study the new use of YL-0919, so the experimental results are credible and provide favorable support for clinical application. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] Figure 1 is the frequency of epileptic seizures in epileptic mice treated with YL-0919; *p<0.05, **p<0.01, ***p<0.001; Figure 2 This is the epileptic seizure classification diagram after YL-0919 treatment of epileptic mice; *p<0.05, **p<0.01, ***p<0.001; Figure 3 This is the analysis of inflammatory factors in the hippocampus of epileptic mice after YL-0919 treatment; *p<0.05, **p<0.01, ***p<0.001; Figure 4 This is the HE staining analysis of the hippocampus of epileptic mice after YL-0919 treatment; Figure 5 This is a Nissl staining analysis of the hippocampus of epileptic mice after YL-0919 treatment. DETAILED DESCRIPTION

[0037] The present invention is further described below in conjunction with specific embodiments, which are only used to explain the present invention and are not to be construed as limiting the present invention. It will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to these embodiments without departing from the principles and purposes of the present invention, and that the scope of the present invention is defined by the claims and their equivalents. The experimental methods for which specific conditions are not specified in the following embodiments are usually tested under conventional conditions or under conditions recommended by the manufacturer.

[0038] Example Study on the therapeutic effect of YL-0919 on epilepsy mouse model 1. Experimental Materials 1.1 Experimental animals 6-8 week old male C57BL / 6J mice were purchased from Beijing SpEd Biotechnology Co., Ltd.

[0039] 1.2 Drugs and reagents TRIzol was purchased from Invitrogen, USA; First Strand cDNA synthesis kit was purchased from Beijing Quanshijin Biotechnology Co., Ltd.; kainate (KA) and YL-0919 were purchased from MCE, USA.

[0040] 1.3 Experimental instruments High-speed cryogenic centrifuge and high-speed cryogenic tissue grinder were purchased from Wuhan Sewell Biotechnology Co., Ltd.; LightCycler480Ⅱ was purchased from Roche, Switzerland; microinjectors and brain stereotaxic apparatus were purchased from Shenzhen Reward Life Science Co., Ltd.

[0041] 2. Experimental Methods 1. Construction of a mouse model of epilepsy induced by acute kainate (KA) The experimental mice were randomly divided into 3 groups. ① Sham group: the same amount of saline was injected into the hippocampus of mice; ② KA group: 1 μL (0.3 ug) KA was injected into the hippocampus of mice; ③ KA+YL-0919 group: 1 μL KA was injected into the hippocampus of mice, and YL-0919 was injected intraperitoneally at the same time.

[0042] The modeling process is as follows: 1) Anesthesia: Mice were anesthetized with 1% sodium pentobarbital (10 μL / g) injected intraperitoneally. The tail of the mouse was pinched with fingers. If there was no response, the anesthesia was successful.

[0043] 2) Fixation: Place the mouse on a digital stereotaxic instrument, hang the mouse's teeth on the front incisor clamp of the digital stereotaxic instrument, keep the mouse's head and body in a straight line and on a horizontal plane, insert the fixing rods on both sides into the ears of the mouse, fix both sides so that the head cannot shake, adjust the mouse's head to the center position, pull the mouse's tongue outward to avoid blocking the airway, and always pay attention to the mouse's condition.

[0044] 3) Skin preparation: Use curved scissors to cut the mouse hair 1 cm on both sides of the sagittal line and disinfect with iodine.

[0045] 4) Expose the bregma: Cut the skin along the sagittal line to an opening of about 2 cm, and the bregma will be visible. Wipe the bregma with 1% hydrogen peroxide and remove the periosteum.

[0046] 5) Positioning: Place a microsyringe containing 1 μL of KA on the digital stereotaxic apparatus, move the syringe needle to the bregma point, take the bregma point as the origin, move the needle tip so that it is 1.8 mm behind the bregma point and 2 mm beside the sagittal line (to the right), and mark the point with a marker.

[0047] 6) Drilling: Use an electric micro handheld cranial drill to drill the skull at the marked point and stop after the dura mater is exposed.

[0048] 7) Brain microinjection: Move the needle of the microinjector back to the marked point and advance the needle at a speed of 1 mm / min. Stop advancing the needle about 2.3 mm into the hippocampus and inject 1 μL of KA or saline into the hippocampus at a speed of 0.02 μL / min. Use a cotton ball dipped in saline to keep the mouse scalp moist. After the injection, leave the needle in place for 5 minutes and slowly pull out the needle to prevent the injection solution from flowing back.

[0049] 8) Skin suture: After observing that there is no active bleeding, suture the mouse scalp and disinfect it with iodine.

[0050] Evaluation of epileptic seizure grading in mice: One hour after intracerebroventricular injection of KA, the mice were evaluated for epileptic seizures according to the Racine scale. Racine scale evaluation criteria: Grade 0: no convulsions; Grade 1: mouth and facial convulsions including jaw clonus; Grade 2: Grade 1 plus rhythmic nodding; Grade 3: Grade 2 plus unilateral forelimb convulsions; Grade 4: bilateral forelimb convulsions with standing; Grade 5: generalized tonic-clonic convulsions with standing falls. Continuous occurrence of Grade 4 or above is considered as status epilepticus (≥2 times in 30 minutes).

[0051] 2. Detection of epileptic seizure frequency in mice After KA was injected into the lateral ventricle of mice, the epileptic seizure status of the mice was continuously recorded within 24 hours using a video monitoring and acquisition system, with the presence of a seizure being recorded as 1 and the absence of a seizure being recorded as 0.

[0052] 3. Detection of inflammatory factor expression in mouse hippocampus tissue After the mice were anesthetized and cardiac perfusion was performed, the hippocampus tissue of the mice was carefully removed, TRIzol was added, and the tissue was homogenized using a tissue grinder, and tissue RNA was extracted to examine the expression of inflammatory factors using the qPCR method.

[0053] 4. HE staining of mouse hippocampus tissue After the mice were anesthetized and cardiac perfusion was performed, the hippocampus tissue of the mice was carefully peeled off. The brain tissue of the mice was fixed with 4% paraformaldehyde solution for 48 hours. After dehydration-embedding-frozen section-hematoxylin staining-differentiation-blueing-eosin staining-dehydration-sealing, the slices were observed under an optical microscope and photographed for preservation.

[0054] 5. Nissl staining of mouse hippocampal tissue After the mice were anesthetized and cardiac perfusion was performed, the hippocampus tissue of the mice was carefully peeled off. The brain tissue of the mice was fixed with 4% paraformaldehyde solution for 48 hours. After dehydration-embedding-frozen section-toluidine blue staining-differentiation-dehydration-sealing, the samples were observed under an optical microscope and photographed for preservation.

[0055] 3. Experimental Results 1. YL-0919 can reduce the frequency of epileptic seizures induced by kainate (KA) in mice The epileptic seizure status of mice was continuously recorded for 24 hours using a video monitoring and acquisition system. Figure 1 As shown, compared with the Sham group, the frequency of epileptic seizures in the KA group was increased, and there was a significant difference in the frequency of epileptic seizures between the KA+YL-0919 group and the KA group, which indicated that YL-0919 significantly reduced the frequency of epileptic seizures induced by kainate (KA) in mice.

[0056] 2. YL-0919 can reduce the grade of epileptic seizures induced by kainate (KA) in mice The Racine scale was used to evaluate the seizures of mice, where seizures of grades one to three were considered focal seizures and grades four to five were considered generalized seizures. Figure 2 As shown, compared with the Sham group, the epileptic behavior of the KA group reached grade 4, and the epilepsy level of the KA+YL-0919 group was reduced by one-fold compared with the KA group, indicating that YL-0919 can reduce the grade of epileptic seizures induced by kainate (KA) in mice.

[0057] 3. YL-0919 can reduce the inflammation level in the hippocampus of epileptic mice induced by kainate (KA) After the mice were anesthetized and cardiac perfusion was performed, the hippocampal tissue of the mice was carefully removed. TRIzol was added and homogenized using a tissue grinder. Tissue RNA was extracted and the expression of inflammatory factors was examined by qPCR. The degree of inflammation in the hippocampus of mice was evaluated using inflammatory factors in the hippocampus (IL-1β, IL-6, TNF-α). Figure 3 As shown, YL-0919 can reduce the degree of inflammation in the hippocampus of epileptic mice induced by kainate (KA).

[0058] 4. YL-0919 can reduce the pathological damage of hippocampal CA1 and CA3 regions in epilepsy induced by kainate (KA) like Figure 4 As shown, HE staining showed that YL-0919 could reduce the pathological damage of CA1 and CA3 regions of the hippocampus in mice with epileptic seizures induced by kainate (KA).

[0059] 5. YL-0919 can reduce the neuronal damage in the CA1 and CA3 regions of the hippocampus induced by kainate (KA) epilepsy like Figure 5 As shown, Nissl staining showed that YL-0919 could reduce the neuronal damage in the CA1 and CA3 regions of the hippocampus in mice induced by kainate (KA) epileptic seizures.

[0060] The description of the above embodiments is only used to understand the method and core idea of ​​the present invention. It should be pointed out that, for those skilled in the art, several improvements and modifications can be made to the present invention without departing from the principle of the present invention, and these improvements and modifications will also fall within the scope of protection of the claims of the present invention.

Claims

1. A drug for treating epilepsy or brain damage caused by epilepsy, characterized in that: The drug comprises an effective dose of a 5HT1A receptor agonist or a pharmaceutically acceptable salt thereof; Preferably, the 5HT1A receptor agonist is YL-0919.

2. The drug according to claim 1, characterized in that The dosage form of the drug includes tablets, capsules, granules, pills, pellets, syrups, powders, granules, suppositories, drops, emulsions, injections, solutions or suspensions.

3. The drug according to claim 1, characterized in that The administration methods of the drug include subcutaneous injection, intradermal injection, intramuscular injection, intraperitoneal injection, intravenous injection, and oral administration; preferably, the administration method of the drug is intraperitoneal injection.

4. Use of a 5HT1A receptor agonist or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating epilepsy or brain damage caused by epilepsy.

5. The use according to claim 4, characterized in that The 5HT1A receptor agonist is YL-0919.

6. The use according to claim 4, characterized in that The effect of the drug is to reduce the frequency of epileptic seizures or reduce the grade of epileptic seizures.

7. The use according to claim 4, characterized in that The effect of the drug is to reduce the degree of inflammation in the hippocampus.

8. The use according to claim 4, characterized in that The effect of the drug is to reduce neuronal damage in the hippocampus.

9. The use according to claim 4, characterized in that: The epilepsy includes primary epilepsy, secondary epilepsy and cryptogenic epilepsy.

10. The use according to claim 9, characterized in that The epilepsy is primary epilepsy; preferably, the epilepsy is temporal lobe epilepsy.

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