Use of YL-0919 in the treatment of epilepsy or epilepsy-induced brain injury

Abstract

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

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of biological medicine, and in particular, the present application relates to a new use of a known compound, and more particularly, the present application relates to the application of YL-0919 in the treatment of epilepsy or epilepsy-induced brain injury. BACKGROUND

[0002] Epilepsy is a chronic brain disease characterized by repeated seizures, which can occur suddenly without any reason, and is one of the most common chronic neurological diseases. The incidence rate of epilepsy in the Chinese population is between 5% and 7%, and the pathogenesis of epilepsy is complex, which has a great impact on the life of patients. Although various antiepileptic drugs can be used for symptomatic treatment of epilepsy, about 1 / 3 of patients are drug-resistant epilepsy.

[0003] Hydroxydipiperonyl ketone (YL-0919) is a new type of small molecule antidepressant, and it has significant antidepressant and anxiolytic effects in various animal models. Studies have shown that YL-0919 is a new type of 5HT1A receptor partial agonist. However, there is still a blank in the research of YL-0919 in the field of treatment of epilepsy or epilepsy-induced brain injury. SUMMARY

[0004] The purpose of the present application is to propose a new use of YL-0919 in a drug for treating epilepsy or epilepsy-induced brain injury, so as to make up for the shortcomings of the prior art.

[0005] The above inventive purpose of the present application is achieved by the following technical solutions:

[0006] In a first aspect, the present application provides a drug for treating epilepsy or epilepsy-induced brain injury.

[0007] Further, the drug comprises an effective dose of a 5HT1A receptor agonist YL-0919 or a pharmaceutically acceptable salt thereof.

[0008] In the present application, the 5HT1A receptor partial agonist includes but is not limited to Buspirone, Tandospirone, Ipsapirone, Aripiprazole, and YL-0919.

[0009] Further, the 5HT 1A receptor agonist is YL-0919.

[0010] Further, the drug can be prepared into a dosage form including tablets, capsules, granules, pills, dripping pills, syrup, powder, powder, suppositories, drops, emulsions, injection solutions, solutions, or suspensions.

[0011] In the present invention, the "YL-0919" is a novel compound with a new structure, which is in powder form, and has a molecular formula of C 18 H 22 YL-0919, with a molecular weight of 334.84, is a 5HT1A receptor agonist and 5-HT uptake blocker, which shows significant antidepressant effects in animal models. YL-0919 has been proven to exert rapid antidepressant and anxiolytic effects, enhance cognitive behavior in mice, and improve ischemic stroke in rats. There is no relevant research or report on the use of YL-0919 for the treatment of epilepsy or brain damage caused by epilepsy, and the present invention is the first report.

[0012] In the present invention, the "effective dose" is supported by experimental research data, which is a proper dose without toxicity to patients, but does not rule out that in some cases, a larger dose that may cause signs of toxicity is forced to be given according to the severity of the disease. The "effective dose" refers to the amount of YL-0919 of the present invention: it is sufficient to cause the prevention of the development or onset of epilepsy or one or more symptoms thereof, to enhance or improve the effect of another treatment, and / or to alleviate one or more symptoms of epilepsy. For subjects suffering from epilepsy, the preferred therapeutically effective amount is an amount effective to reduce the frequency of seizures and / or reduce hippocampal neuronal pathological damage.

[0013] In some embodiments, a therapeutically effective amount can be administered to a patient in one or more doses sufficient to reduce, alleviate, stabilize, reverse or slow the progression of the disease, or otherwise lessen the pathological consequences of the disease, or lessen the symptoms of the disease. The reduction or alleviation need not be permanent, but can be for a range of time (at least an hour, at least a day, or at least a week or more). Effective amounts are typically determined by a physician, on a case-by-case basis, and are within the skill of the art. Several factors are typically taken into account when determining the appropriate dose to achieve an effective amount. These factors include the patient's age, sex, and weight, the condition being treated, the severity of the condition, and the route, dosage form and regimen of administration, and the desired result.

[0014] In some embodiments, the dose administered depends on many factors such as the nature and severity of the disease to be prevented and / or treated (in the present invention, in particular epilepsy or brain damage caused by epilepsy), the gender, 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-mentioned dose can be administered in a single dose form or divided into several, for example two, three or four, dose forms.

[0015] Further, various types of formulations can be made depending on the formulation method, administration method, age, body weight, sex, disease state, diet, administration time, administration route, excretion rate, and response sensitivity of the patient, and a skilled medical doctor can easily determine the formulation and the administration dose effective for the desired prophylaxis and / or treatment.

[0016] The term "pharmaceutically acceptable salt" refers to a pharmaceutically acceptable salt of YL-0919, which means those carboxylic acid salts, amino acid addition salts, and the like of the compounds described herein (YL-0919) that are, within the scope of sound medical judgment, suitable for use in contact with the patients without producing any undesirable toxicity, irritation, allergic response, and the like, commensurate with a reasonable benefit / risk ratio, and effective for their intended applications, including, where possible, the zwitterionic forms of the compounds described herein (YL-0919).

[0017] In some embodiments, examples of the "pharmaceutically acceptable salt of YL-0919" include, but are not limited to, a salt with an alkali metal ion such as Li + , Na + , or K + , or a salt with an alkaline earth metal ion such as Mg 2+ or Ca 2+ , or a salt with any other pharmaceutically acceptable metal ion such as Zn 2+ or Al 3+ , or a pharmaceutically acceptable salt with an organic base such as diethanolamine, ethanolamine, N-methylglucamine, triethanolamine, or tromethamine.

[0018] In some embodiments, the pharmaceutically acceptable base addition salts are formed between metal or amine ions, for example, alkali and alkaline earth metal hydroxides or organic amines. Examples of metals which can be used as cations are sodium, potassium, magnesium, calcium, and the like. Examples of suitable amines are N,N'-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, N-methylglucamine, and procaine.

[0019] In some embodiments, the base addition salts of acidic compounds can be prepared by contacting the free acid form with a sufficient amount of the desired base to produce the salt. The free acid form can be regenerated by contacting the salt form with a sufficient amount of an acid to produce the free acid form. The free acid and base forms can be regenerated by in vitro

[0020] In some embodiments, the salt can be a sulfate, bisulfate, hydrogen sulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide, acid such as hydrochloric acid, nitric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, phosphoric acid, and the like prepared from inorganic acids. Representative salts include the following: hydrobromide, hydrochloride, sulfate, hydrogen sulfate, nitrate, acetate, oxalate, valerate, oleate, palmitate, stearate, laurate, borate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, naphthoate, mesylate, glucoheptonate, lactobionate, laurylsulfoacetate, and isethionate, and the like. The salt can also be prepared from organic acids such as aliphatic mono- and dicarboxylic acids, phenyl-substituted alkanoic acids, hydroxy alkanoic acids, alkyldioic acids, aromatic acids, aliphatic and aromatic sulfonic acids, etc. Representative salts include acetate, propionate, caprylate, isobutyrate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, mandelate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, naphthoate, benzenesulfonate, toluenesulfonate, phenylacetate, citrate, lactate, maleate, tartrate, mesylate, and the like. Pharmaceutically acceptable salts can include cations based on alkali and alkaline earth metals, for example sodium, lithium, potassium, calcium, magnesium, and aluminum, as well as ammonium, quaternary ammonium, and amine cations such as, but not limited to ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine, and the like. Salts of amino acids such as arginate, gluconate, galacturonate, and the like, are also encompassed by the term.

[0021] The administration of the drug YL-0919 of the present application includes subcutaneous injection, intradermal injection, intramuscular injection, intraperitoneal injection, intravenous injection, oral administration, etc. In previous studies on beagle dogs, gavage administration was used, and in studies on cerebral stroke in rats, intraperitoneal injection was used. The present application preferably uses intraperitoneal injection to improve the stability of the drug in mice.

[0022] In a second aspect, the present application provides the use of a 5HT1A receptor agonist or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating epilepsy or brain injury caused by epilepsy.

[0023] The 5HT1A receptor partial agonist of the present application includes, but is not limited to, Buspirone, Tandospirone, Ipsapirone, Aripiprazole, YL-0919, etc.

[0024] Further, the 5HT1A receptor agonist is YL-0919.

[0025] Further, the effect of the drug is to reduce the frequency of seizures.

[0026] Further, the effect of the drug is to reduce the inflammation degree of hippocampus.

[0027] Further, the effect of the drug is to reduce the pathological damage of hippocampus neurons.

[0028] Further, the epilepsy includes primary epilepsy, secondary epilepsy, and cryptogenic epilepsy.

[0029] Preferably, the epilepsy is secondary epilepsy.

[0030] More preferably, the secondary epilepsy is temporal lobe epilepsy.

[0031] The causes of the epilepsy according to the present application include, but are not limited to, muscle contraction, brain injury, genetic factors, physical illness, drug abuse, etc. The epilepsy caused by brain injury is the most important cause of seizures.

[0032] The establishment of the epilepsy or brain injury caused by the epilepsy according to the present application requires the construction of a mouse model. The construction method of the epilepsy mouse model includes, but is not limited to, the kindling model, the pentylenetetrazol (PTZ) epilepsy model, the kainic acid (KA) epilepsy model, the maximal electroshock (MES) model, the gene knockout and transgenic epilepsy animal model. The kainic acid (KA) epilepsy model is preferred according to the present application, because the KA model is more similar to humans in terms of latency, behavioral symptoms, duration of latency, and electroencephalogram characteristics in the latency and chronic phases, and is more valuable for research.

[0033] The judgment method of the epilepsy or brain injury caused by the epilepsy according to the present application involves multiple aspects, including but not limited to the observation of neurobehavioral characteristics, the analysis of brain tissue morphology, the spongy change, the density of neuron dendritic spines, the expression of inflammation-related genes, the detection of inflammation protein expression, the electroencephalogram analysis, the brain blood flow analysis, the inflammation degree of hippocampus, the pathological damage of neurons, etc. The analysis of the inflammation degree of hippocampus and the pathological damage of neurons according to the present application is an important indicator for judging the brain injury of mice.

[0034] The secondary epilepsy caused by brain injury includes, but is not limited to, temporal lobe epilepsy, epilepsy caused by brain tumors, cerebral hemorrhage, cerebral infarction, viral encephalitis, autoimmune encephalitis, and focal cortical dysplasia, etc. The temporal lobe epilepsy is the most common secondary epilepsy according to the present application, and is also the most common type of brain injury caused by epilepsy.

[0035] Advantages and beneficial effects of the present application:

[0036] The application first finds that YL-0919 can be used for treating epilepsy or brain injury caused by epilepsy, and provides a new angle and method for clinical treatment of epilepsy or brain injury caused by epilepsy.

[0037] The application uses a mouse model to study the new use of YL-0919, so that the experimental results are reliable and provide favorable support for clinical application. BRIEF DESCRIPTION OF DRAWINGS

[0038] Figure 1 is a seizure frequency graph of a mouse with epilepsy treated by YL-0919; *p<0.05, **p<0.01, ***p<0.001;

[0039] Figure 2 is a seizure classification graph of a mouse with epilepsy treated by YL-0919; *p<0.05, **p<0.01, ***p<0.001;

[0040] Figure 3 is an analysis graph of inflammatory factors in the hippocampus of a mouse with epilepsy treated by YL-0919; *p<0.05, **p<0.01, ***p<0.001;

[0041] Figure 4 is an analysis graph of HE staining in the hippocampus of a mouse with epilepsy treated by YL-0919;

[0042] Figure 5 is an analysis graph of Nissl staining in the hippocampus of a mouse with epilepsy treated by YL-0919. DETAILED DESCRIPTION

[0043] The application will be further described below in conjunction with specific embodiments, which are only used to explain the application and cannot be understood as a limitation of the application. Those skilled in the art can understand that various changes, modifications, replacements and variations can be made to the embodiments without departing from the principles and purposes of the application, and the scope of the application is defined by the claims and their equivalents. The experimental methods in the following examples are not specified, and the detection is usually carried out according to the conventional conditions or the conditions recommended by the manufacturer.

[0044] Example YL-0919 treatment effect research on a mouse model with epilepsy

[0045] I. Experimental materials

[0046] 1.1 Experimental animals

[0047] 6-8-week-old male C57BL / 6J mice were purchased from Sibeifu Beijing Biotechnology Co., Ltd.

[0048] 1.2 Drugs and reagents

[0049] TRIzol was purchased from Invitrogen, USA; First Strand cDNA synthesis kit was purchased from Beijing Zison Biotechnology Co., Ltd.; Kainic acid (KA) and YL-0919 were purchased from MCE, USA.

[0050] 1.3 Experimental instruments

[0051] High-speed low-temperature centrifuge and high-speed low-temperature tissue grinder were purchased from Wuhan Seville Biotechnology Co., Ltd.; LightCycler480Ⅱ was purchased from Roche, Switzerland; microsyringe and brain stereotaxic apparatus were purchased from Shenzhen Ruivode Life Science and Technology Co., Ltd.

[0052] II. Experimental methods

[0053] 1. Construction of kainic acid (KA) acute induced epilepsy mouse model

[0054] The experimental mice were randomly divided into 3 groups. ① Sham group: the mice were injected with the same amount of normal saline into the hippocampus; ② KA group: the mice were injected with 1 μL (0.3 ug) KA into the hippocampus; ③ KA+YL-0919 group: the mice were injected with 1 μL KA into the hippocampus, and YL-0919 was injected intraperitoneally.

[0055] The modeling process is as follows:

[0056] 1) Anesthesia: the mice were anesthetized by intraperitoneal injection of 1% sodium pentobarbital (10 μL / g), and the mouse tail was pinched with fingers. If there was no response, the anesthesia was successful.

[0057] 2) Fixation: the mouse was placed on the digital brain stereotaxic apparatus, the mouth and teeth were hung on the front door teeth forceps of the digital brain stereotaxic apparatus, the mouse head and body were kept in a straight line and a horizontal plane, the two side fixing rods were inserted into the mouse's ears on both sides, the head was fixed so that it could not move, the mouse's head was adjusted to the center position, the mouse's tongue was pulled out to avoid blocking the airway, and the mouse's state was observed at all times.

[0058] 3) Skin preparation: the scissors were bent along the sagittal line, and the mouse hair was cut off 1 cm on both sides, and iodophor was used for disinfection.

[0059] 4) Expose the skull: cut the skin along the sagittal line, open about 2 cm, and the bregma point can be seen. The bregma point was wiped with 1% hydrogen peroxide, and the periosteum was removed.

[0060] 5) Positioning: the microsyringe containing 1 μL of KA was installed on the digital brain stereotaxic apparatus, the needle of the microsyringe was moved to the bregma point, the bregma point was taken as the origin, and the needle tip was moved to 1.8 mm behind the bregma point and 2 mm to the right of the sagittal line. The point was marked with a marker pen.

[0061] 6) Opening of the skull: Drill the skull at the marked point with a motorized micro handheld skull drill, stop after the dura mater is exposed.

[0062] 7) Microinjection: Move the microinjection needle to the marked point, insert at a speed of 1 mm / min, stop at approximately 2.3 mm, inject at a speed of 0.02 μL / min, inject 1 μL of KA or normal saline into the hippocampus, keep the mouse scalp moist with a cotton ball dipped in normal saline, leave the needle in for 5 min after injection, and slowly pull out the needle to prevent backflow of the injection.

[0063] 8) Suture: After no active bleeding is observed, suture the mouse scalp and disinfect with iodophor.

[0064] Classification of mouse seizure assessment:

[0065] After 1 h of intracerebroventricular injection of KA in mice, the seizure was assessed according to the Racine scale. The evaluation criteria of the Racine scale are as follows: 0: no convulsive seizure; 1: oral and facial convulsions including jaw clonus; 2: rhythmic head nodding based on level 1; 3: unilateral forelimb convulsions based on level 2; 4: bilateral forelimb convulsions with standing; 5: generalized tonic-clonic convulsions with standing and falling, and those with 4 or 5 levels for more than 30 min are determined as status epilepticus (≥2 times).

[0066] 2, Detection of mouse seizure frequency

[0067] After intracerebroventricular injection of KA in mice, the seizure state of the mice was continuously recorded for 24 h by a video monitoring and acquisition system, and the mice with seizures were recorded as 1 and those without seizures were recorded as 0.

[0068] 3, Detection of expression of inflammatory factors in mouse hippocampal tissue

[0069] After heart perfusion under anesthesia, the mouse hippocampal tissue was carefully stripped. TRIzol was added, homogenized using a tissue grinder, and the tissue RNA was extracted. The expression of inflammatory factors was checked by qPCR method.

[0070] 4, Detection of HE staining of mouse hippocampal tissue

[0071] After heart perfusion under anesthesia, the mouse hippocampal tissue was carefully stripped. The mouse brain tissue was fixed with 4% paraformaldehyde solution for 48 h. Through dehydration-embedding-frozen section-hematoxylin staining-differentiation-blueing-erythrosin staining-dehydration-mounting, the tissue was observed under an optical microscope and photographed for preservation.

[0072] 5, Detection of Nissl staining of mouse hippocampal tissue

[0073] After heart perfusion, the mouse brain hippocampus tissue was carefully stripped. The mouse brain tissue was fixed with 4% paraformaldehyde solution for 48 h. By dehydration-embedding-freezing section-methylrosaniline blue staining-differentiation-dehydration-sealing, observed under optical microscope and photographed.

[0074] III. Experimental results

[0075] 1. YL-0919 can reduce the frequency of kainic acid (KA) induced seizures in mice

[0076] The status of mouse seizures within 24 h was continuously recorded by video monitoring acquisition system. The results are shown in Figure 1 Compared with the Sham group, the frequency of seizures in the KA group increased, and there was a significant difference in the frequency of seizures between the KA+YL-0919 group and the KA group, which indicated that YL-0919 significantly reduced the frequency of kainic acid (KA) induced seizures in mice.

[0077] 2. YL-0919 can reduce the classification of kainic acid (KA) induced seizures in mice

[0078] The Racine scale was used to evaluate the seizures in mice, and the first to third seizures were considered to be focal seizures, and the fourth to fifth were considered to be generalized seizures. The results are shown in Figure 2 Compared with the Sham group, the KA group reached the fourth level of seizure behavior, and the KA+YL-0919 group reduced the seizure level by half compared with the KA group, indicating that YL-0919 can reduce the classification of kainic acid (KA) induced seizures in mice.

[0079] 3. YL-0919 can reduce the degree of inflammation in the hippocampus of kainic acid (KA) induced seizure mice

[0080] After heart perfusion, the mouse brain hippocampus tissue was carefully stripped. TRIzol was added, homogenized using a tissue grinder, and the tissue RNA was extracted. The expression of inflammatory factors was checked by qPCR method. The degree of inflammation in the hippocampus of mice was evaluated by hippocampal inflammatory factors (IL-1β, IL-6, TNF-α), as shown in Figure 3 YL-0919 can reduce the degree of inflammation in the hippocampus of kainic acid (KA) induced seizure mice.

[0081] 4. YL-0919 can reduce the pathological damage of hippocampal CA1 and CA3 regions in kainic acid (KA) induced seizure mice

[0082] As shown in Figure 4 HE staining showed that YL-0919 can reduce the pathological damage of hippocampal CA1 and CA3 regions in kainic acid (KA) induced seizure mice.

[0083] 5. YL-0919 can reduce kainic acid (KA)-induced seizure neuronal damage in the hippocampal CA1 and CA3 regions

[0084] As shown in FIG. 5, Nissl staining showed that YL-0919 can reduce kainic acid (KA)-induced seizure neuronal damage in the hippocampal CA1 and CA3 regions of mice. Figure 5

[0085] The above description of the embodiments is only for understanding the method of the present application and its core idea. It should be noted that for those skilled in the art, without departing from the principles of the present application, the present application can be improved and modified in several ways, and these improvements and modifications will also fall within the scope of protection of the claims of the present application.​

Claims

1. The use of YL-0919 or its pharmaceutically acceptable salt in the preparation of medicaments for treating epilepsy or epilepsy-induced brain injury, characterized in that, The drug can reduce the frequency or severity of epileptic seizures, and can also reduce inflammation or neuronal damage in the hippocampus.

2. The use according to claim 1, characterized in that, The epilepsy mentioned includes primary epilepsy, secondary epilepsy, and cryptogenic epilepsy.

3. The use according to claim 2, characterized in that, The epilepsy mentioned is primary epilepsy.

4. The use according to claim 3, characterized in that, The epilepsy in question is temporal lobe epilepsy.

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