Lithium valproate and its crystalline forms, methods of preparation, pharmaceutical compositions and uses

Abstract

The application relates to the technical field of pharmaceutical chemistry, in particular to a valproate lithium, a crystal form thereof, a preparation method, a pharmaceutical composition and application. The crystal form of the valproate lithium comprises crystal form I, crystal form II, crystal form III, crystal form IV, crystal form V, crystal form VI and crystal form VII. The valproate lithium provided by the application has a more comprehensive treatment effect on nervous system diseases, is good in curative effect and good in stability.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of medicine, in particular to a lithium valproate, a crystal form thereof, a preparation method, a pharmaceutical composition and an application. BACKGROUND

[0002] Neurological diseases refer to diseases occurring in the central nervous system, peripheral nervous system and autonomic nervous system, and mainly manifesting as sensory, motor, consciousness and autonomic nervous dysfunction. Neurological diseases can cause epilepsy, mania, splitting, bipolar disorder, anxiety, depression and agitation, etc., leading to impaired motor and sensory functions, cognitive and psychological disorders, and seriously affecting the normal life of patients.

[0003] Epilepsy is a chronic brain disease characterized by repeated seizures, which is caused by abnormal discharge of neurons in the brain. The disease has the characteristics of recurrence and transience. Valproic acid is a kind of antiepileptic drug, which has different degrees of antagonistic effect on convulsions caused by various methods, and is effective for various types of epilepsy such as each type of petit mal, myoclonic epilepsy, focal seizures, grand mal and mixed epilepsy. Sodium valproate is a commonly used antiepileptic drug in clinic, and there are currently sodium valproate ordinary preparation and sustained-release preparation in clinical development. The dosage of sodium valproate sustained-release tablets for treating epilepsy or mania is 500mg / day for adults, taken twice a day, once in the morning and once in the evening, and the maintenance dose is 1000mg-2000mg / day, which means that patients will also intake too much sodium when taking sodium valproate sustained-release tablets.

[0004] Depression, also known as depressive disorder, is a mental disorder with high incidence, high clinical cure rate, low treatment acceptance rate and high recurrence rate. Its main feature is significant and persistent low mood, and patients may have self-injury, suicidal behavior, and psychotic symptoms such as delusion and hallucination. Drug treatment is the main treatment for depression. The main anti-depression drugs in clinical practice are a class of selective serotonin reuptake inhibitors (SSRIs) and serotonin and norepinephrine reuptake inhibitors (SNRIs), etc. These drugs generally take effect within 2-4 weeks. Because the effect is slow, for patients with depression, especially severe patients, adverse reactions appear before the effect appears, and patients often refuse to take the medicine in this 1-2 weeks, and even commit suicide during this period, which is a clinical pain point of anti-depression treatment.

[0005] Agitation is a common acute syndrome in psychiatry, and is a comorbidity feature of many neurological diseases including schizophrenia, anxiety, depression, dementia and Alzheimer's disease. It can manifest as agitation in schizophrenia, psychotic agitation in patients with psychotic disorders, anxiety agitation in severe anxiety, psychomotor agitation in depressive episodes and mixed episodes of bipolar disorder, and agitation in Alzheimer's disease. In clinical practice, agitation is manifested as a series of thinking activities, emotions and behaviors from low to high levels of excitement, and cannot be calm. Patients often show obvious restlessness and excessive physical activity, accompanied by anxiety, and severe agitation can manifest as excitement, threats, attacks, self-harm and other behaviors. The neurobiological mechanism of agitation is not yet clear. Related studies suggest that abnormal enhancement of dopamine and norepinephrine activity in the brain and low gamma-aminobutyric acid activity may be the underlying mechanism of agitation, and abnormal 5-HT (serotonin) activity may also be related to the occurrence of agitation. These neurotransmitter systems regulate and influence each other and play an important role in the development of agitation. There is no FDA-approved effective drug for the treatment of agitation symptoms including schizophrenia agitation, anxiety agitation, depression agitation and Alzheimer's disease agitation. Clinicians have always relied on the off-target effects of existing marketed anti-schizophrenia drugs, anti-anxiety drugs and anti-depression drugs to try to control agitation symptoms. The effectiveness of these drugs in treating agitation is limited. Therefore, there is an urgent need to develop a safe and effective drug for the treatment or control of agitation to improve the overall prognosis of patients. SUMMARY

[0006] Therefore, one or more embodiments of the present application provide a lithium valproate and a crystal form thereof, a preparation method, a pharmaceutical composition and an application. The lithium valproate of the present application has a more comprehensive treatment effect on neurological diseases, good efficacy and good stability. The crystal form of the lithium valproate of the present application has the advantages of not being easy to absorb moisture and good stability.

[0007] The technical solution of the present application includes the following contents:

[0008] The lithium valproate has the following structure:

[0009]

[0010] Further, the lithium valproate is a lithium salt of valproic acid, wherein the chemical ratio of valproic acid to lithium is 1:1.

[0011] Further, the lithium salt of valproic acid is a crystalline salt.

[0012] A preparation method of lithium valproate includes the following steps:

[0013] mixing valproic acid, lithium hydroxide and an aqueous alcohol solution at 0-10℃ to obtain a first solution;

[0014] stirring the first solution at 20-40℃ for 12-20h to obtain a second solution;

[0015] filtering the second solution to collect the solid to obtain the lithium valproate;

[0016] The structure of the lithium valproate is as follows:

[0017]

[0018] A crystal form of the lithium valproate described above is selected from any one of the following groups:

[0019] (1) Crystal Form I: its X-ray diffraction powder diffraction pattern includes characteristic diffraction peaks at the following 2θ (°) angles: 6.710±0.2°, 7.853±0.2°, 21.674±0.2°, 19.067±0.2°, 20.070±0.2°, 23.617±0.2°, 20.445±0.2° and 15.699±0.2°;

[0020] (2) Crystal Form II: its X-ray diffraction powder diffraction pattern includes characteristic diffraction peaks at the following 2θ (°) angles: 6.660±0.2°, 7.503±0.2°, 18.813±0.2°, 19.522±0.2°, 20.058±0.2°, 22.453±0.2°, 22.729±0.2° and 15.093±0.2°;

[0021] (3) Crystal Form III: its X-ray diffraction powder diffraction pattern includes characteristic diffraction peaks at the following 2θ (°) angles: 6.703±0.2°, 7.869±0.2°, 21.636±0.2°, 23.632±0.2°, 19.080±0.2°, 20.054±0.2°, 15.713±0.2° and 27.420±0.2°;

[0022] (4) Crystal Form IV: its X-ray diffraction powder diffraction pattern includes characteristic diffraction peaks at the following 2θ (°) angles: 6.753±0.2°, 7.883±0.2°, 19.169±0.2°, 21.880±0.2°, 20.263±0.2°, 23.716±0.2°, 15.755±0.2° and 10.930±0.2°;

[0023] (5) Form V: its X-ray diffraction powder pattern includes the following characteristic diffraction peaks at 2-theta (°) angles: 6.741±0.2°, 7.914±0.2°, 21.680±0.2°, 23.673±0.2°, 19.134±0.2°, 20.094±0.2°, 15.756±0.2° and 10.847±0.2°;

[0024] (6) Form VI: its X-ray diffraction powder pattern includes the following characteristic diffraction peaks at 2-theta (°) angles: 6.784±0.2°, 7.946±0.2°, 21.743±0.2°, 19.152±0.2°, 23.707±0.2°, 20.502±0.2°, 20.150±0.2° and 21.164±0.2°; and,

[0025] (7) Form VII: its X-ray diffraction powder pattern includes the following characteristic diffraction peaks at 2-theta (°) angles: 6.729±0.2°, 7.702±0.2°, 18.991±0.2°, 20.702±0.2°, 21.741±0.2°, 22.051±0.2°, 22.533±0.2° and 28.957±0.2°.

[0026] Further, the crystal form of the lithium valproate has at least one of the following characteristics:

[0027] (1) the differential scanning calorimetry curve of the Form I has an endothermic peak at 359.2±3°C;

[0028] (2) the differential scanning calorimetry curve of the Form II has an endothermic peak at 361.8±3°C;

[0029] (3) the differential scanning calorimetry curve of the Form III has an endothermic peak at 331.8±3°C;

[0030] (4) the differential scanning calorimetry curve of the Form V has an endothermic peak at 357.2±3°C;

[0031] (6) the differential scanning calorimetry curve of the Form VI has an endothermic peak at 362.3±3°C; and,

[0032] (7) the differential scanning calorimetry curve of the Form VII has an endothermic peak at 362.2±3°C.

[0033] A method for preparing a crystal form of the lithium valproate described above, comprising the following steps:

[0034] The lithium valproate is crystallized by at least one of the following methods: solution crystallization, anti-solvent crystallization, evaporation crystallization, volatile crystallization and melt crystallization; wherein the solvent used includes at least one of the following: ethanol, dichloromethane, acetonitrile, ethyl acetate, tetrahydrofuran, methanol, petroleum ether, butyl acetate, methyl tert-butyl ether, n-butanol, dimethyl sulfoxide, tert-butyl alcohol, isopropyl alcohol, toluene and N,N-dimethylformamide.

[0035] A pharmaceutical composition, characterized in that the pharmaceutical composition comprises a pharmaceutical active ingredient and at least one of a pharmaceutically acceptable excipient and a carrier;

[0036] The pharmaceutical active ingredient comprises lithium valproate;

[0037] The structure of the lithium valproate is as follows:

[0038]

[0039] Use of the lithium valproate or the pharmaceutical composition described above in the preparation of a medicament for the treatment and / or prevention of a nervous system disease.

[0040] Further, the nervous system disease comprises at least one of epilepsy, mania, schizotypal, bipolar disorder, anxiety, depression and agitation.

[0041] The advantages of the present application over the prior art include at least the following aspects:

[0042] (1) Sodium valproate and lithium carbonate are commonly used as an antiepileptic drug in clinic, and the maintenance dose in treatment is 1000mg-2000mg / day. The use of a large dose often shows related adverse reactions, and the blood lithium concentration needs to be detected during the clinical use of lithium carbonate, and the sodium content is high during the clinical use of sodium valproate. Magnesium valproate can be used to avoid high sodium intake of sodium valproate. However, the effect of magnesium valproate is slow after administration, and the action time is long. The lithium valproate is designed and successfully synthesized in the present application. On the one hand, high salt intake is avoided, and on the other hand, the lithium valproate has a better effect than lithium carbonate or sodium valproate at the same dose, and has a treatment and / or prevention effect on nervous system diseases at a low dose compared with sodium valproate and magnesium valproate.

[0043] (2) The crystal form of sodium valproate disclosed at present shows high hygroscopicity. High hygroscopicity can cause the drug to absorb water during storage, thereby causing the chemical or physical stability of the drug to decrease, and affecting the shelf life of the drug. The crystal form of lithium valproate with low hygroscopicity is developed in the present application, which ensures a lower water content of the product, and makes the lithium valproate more stable in quality during storage.

[0044] (3) The current clinical mainstream antidepressants include selective serotonin reuptake inhibitors (SSRIs) and serotonin and norepinephrine reuptake inhibitors (SNRIs), which generally need to start to take effect in 2-4 weeks. Since the effect is slow, for patients with depression, especially severe patients, adverse reactions appear before the therapeutic effect appears. The valproate lithium of the present application can quickly take effect and can play a therapeutic effect in 8-10 days, effectively improving patient compliance and improving therapeutic effect.

[0045] (4) The valproate lithium of the present application has an improvement effect on the manifestations of various nervous system diseases, including epilepsy, mania, schizophrenia, bipolar disorder, anxiety, depression, and agitation. Agitation is a common acute syndrome in psychiatry, and is a comorbidity feature of various neurological diseases including schizophrenia, anxiety, depression, dementia, and Alzheimer's disease. There is currently no effective drug for treating symptoms including schizophrenia agitation, anxiety agitation, depression agitation, and Alzheimer's disease agitation. Clinically, off-target effects of existing marketed anti-schizophrenia drugs, anti-anxiety drugs, and anti-depression drugs are often used to try to control agitation symptoms, but the effect is very limited. The valproate lithium of the present application has a significant anti-agitation effect and can be used to prepare a drug for treating and / or preventing agitation. The anti-agitation mechanism of the valproate lithium of the present application includes delaying the occurrence of hyperactivity state caused by MK-801 and reducing the degree of hyperactivity caused by MK-801. BRIEF DESCRIPTION OF DRAWINGS

[0046] In order to more clearly illustrate the specific embodiments of the present application or the technical solutions in the prior art, the drawings needed in the specific embodiments or prior art description will be briefly introduced below. Obviously, the drawings in the following description are some embodiments of the present application, and those skilled in the art can also obtain other drawings according to these drawings without creative labor.

[0047] Figure 1 XPRD pattern of the crystal form I of the valproate lithium prepared for Example 2 of the present application;

[0048] Figure 2 XPRD pattern of the crystal form II of the valproate lithium prepared for Example 2 of the present application;

[0049] Figure 3 XPRD pattern of the crystal form III of the valproate lithium prepared for Example 2 of the present application;

[0050] Figure 4 XPRD pattern of the crystal form IV of the valproate lithium prepared for Example 2 of the present application;

[0051] Figure 5XPRD pattern of the crystal form V of the lithium valproate prepared in Example 2 of the present application;

[0052] Figure 6 XPRD pattern of the crystal form VI of the lithium valproate prepared in Example 2 of the present application;

[0053] Figure 7 XPRD pattern of the crystal form VII of the lithium valproate prepared in Example 2 of the present application;

[0054] Figure 8 Graph of the body weight change of the mice during the modeling of the mouse depression model of Example 5 of the present application (***p<0.001, compared with the normal group);

[0055] Figure 9 Graph of the immobility time in the tail suspension test of the mouse depression model of Example 5 of the present application (***p<0.001, compared with the normal group);

[0056] Figure 10 Graph of the data of the open field test of the mouse depression model of Example 5 of the present application (***p<0.05, compared with the normal group);

[0057] Figure 11 Graph of the sugar water preference rate affected by the tested drug of the mouse depression model of Example 5 of the present application (**p<0.01, compared with the normal group);

[0058] Figure 12 Graph of the immobility time in the tail suspension test affected by the tested drug of the mouse depression model of Example 5 of the present application;

[0059] Figure 13 Statistical results of the forced swimming behavior affected by the tested drug of the mouse depression model of Example 5 of the present application;

[0060] Figure 14 Statistical results of the open field behavior affected by the tested drug of the mouse depression model of Example 5 of the present application (***p<0.001, compared with the normal group);

[0061] Figure 15 Statistical results of the effect of the lithium valproate of Example 6 of the present application on the movement distance of the model animals, Figure 15 A is the movement distance of each group of mice within 10 min after injection of MK-801 during the test, and the movement distance of the mice within 12 10 min is recorded, i.e. a fold line graph of the movement distance of the mice within 12 10 min, Figure 15 B is the total movement distance of each group of mice within 120 min. DETAILED DESCRIPTION

[0062] The application will be further described below with reference to the embodiments and examples. It should be understood that these examples are only used to illustrate the application and not intended to limit the scope of the application. Furthermore, it should be understood that those skilled in the art can make various modifications or changes to the application after reading the content taught in the application, and these equivalent forms also fall within the protection scope of the claims of the application.

[0063] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used in the description of the application herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the application.

[0064] The selection range of the term "and / or" used herein includes any one of two or more related listed items, and also includes any and all combinations of the related listed items, which includes any two related listed items, any more related listed items, or a combination of all related listed items. It should be noted that when at least three items are connected by at least two conjunctions selected from "and / or", "or / and", and "and / or", it should be understood that in the application, the technical solution undoubtedly includes the technical solution connected by "logical and", and also undoubtedly includes the technical solution connected by "logical or". For example, "A and / or B" includes three parallel solutions of A, B, and A+B.

[0065] In this application, "preferably", "more preferably", "even more preferably" and the like are merely used to describe embodiments or examples with better effects, and it should be understood that they do not constitute a limitation on the protection scope of the application.

[0066] In this application, "further" is used to describe the difference in content, but should not be understood as a limitation on the protection scope of the application.

[0067] In this application, the technical features described in an open form include both the closed technical solution consisting of the listed features and the open technical solution containing the listed features.

[0068] In this application, with respect to the numerical interval (i.e. numerical range), if not specifically stated, the optional numerical distribution within the above numerical interval is considered to be continuous, and includes the two numerical end points (i.e. the minimum value and the maximum value) of the numerical range, and each numerical value between the two numerical end points. If not specifically stated, when the numerical interval only points to the integers within the numerical interval, it includes the two end point integers of the numerical range, and each integer between the two end points. In addition, when multiple ranges are provided to describe a feature or a characteristic, these ranges can be combined. In other words, unless otherwise indicated, the ranges disclosed herein should be understood to include any and all sub-ranges therein.

[0069] The temperature parameters in the present application, if not specifically defined, allow for constant temperature treatment as well as for variations within a certain temperature interval. It is to be understood that constant temperature treatment allows for fluctuations within the accuracy of the instrument control. Fluctuations within a range such as ± 0.5°C, ± 0.4°C, ± 0.3°C, ± 0.2°C, ± 0.1°C are allowed.

[0070] Room temperature in the present application generally refers to 20°C to 40°C.

[0071] In the present application, weight can be μg, mg, g, kg and other mass units well known in the chemical industry.

[0072] Compound of the present application

[0073] In the present application, compound refers to valproate lithium.

[0074] The structural formula of valproate lithium is:

[0075] The crystalline forms of valproate are Form I, Form II, Form III, Form IV, Form V, Form VI and Form VII.

[0076] Crystallization

[0077] Crystallization on production scale can be accomplished by manipulating the solution so that the solubility limit of the compound is exceeded. This can be done by a variety of methods, for example, dissolving the compound at a relatively high temperature and then cooling the solution below the saturation limit or by reducing the volume of the liquid by boiling, evaporation at atmospheric pressure, drying under vacuum or by some other method. The solubility of the compound can be reduced by adding an antisolvent or a solvent in which the compound has a low solubility or a mixture of such solvents. Another alternative is to adjust the pH to reduce the solubility. For a detailed description of crystallization see Crystallization, Third Edition, J W Mullens, Butterworth Heineman Ltd. 1993, ISBN 0750611294.

[0078] Identification and properties of the crystalline forms

[0079] The properties of the compounds of the present application were investigated after their preparation in different crystalline forms using a variety of methods and instruments as described below.

[0080] X-ray powder diffraction (XRPD)

[0081] Methods for determining the X-ray powder diffraction of crystalline forms are known in the art. XRPD can detect information of changes in crystalline form, crystallinity, crystal structure state, etc. and is a common means for identifying crystalline forms. The peak positions of XRPD patterns are mainly dependent on the structure of the crystalline form, and the measurement of 2Θ of XRPD patterns can be slightly different between different instruments, so the values of 2Θ cannot be considered absolute. According to the instrument used in the experiments of the present application, there is an error of ±0.2° in the diffraction peak. It can be understood that the range of error is not absolute for different test instruments and test conditions. The crystalline form of the compound of the present application has a specific crystal form and has specific characteristic peaks in the XRPD pattern.

[0082] In the present application, "the X-ray powder diffraction pattern has a characteristic diffraction peak at a specific 2Θ angle" means that the peak value of the peak is within the indicated numerical range, near the indicated numerical point, or near the indicated numerical range.

[0083] Due to differences in measurement instruments and measurement conditions, the actual obtained X-ray powder diffraction pattern of some peaks or peak values may be slightly shifted, that is, there may be slight differences between the characteristic peak combination or X-ray powder diffraction pattern indicated in the present application. However, it can be understood that for those skilled in the art, whether the slightly different characteristic peak combination or X-ray powder diffraction pattern can be substantially constituted as the crystalline form of the present application can be identified as a whole. Therefore, these that are substantially recognized as consistent with the crystalline form of the present application should be considered within the protection scope of the present application.

[0084] For example, "the X-ray powder diffraction pattern has a peak at a diffraction angle of 6.710±0.2°" means that the peak value can be within the range of 6.710±0.2° and near the range, as long as it does not affect the overall identification of the crystalline form. In addition, "±0.2" here only represents the error of the peak value in the diffraction angle position, and is irrelevant to the peak shape and peak width of the peak.

[0085] In the present application, "substantially" in "the X-ray powder diffraction pattern is substantially as characterized by a specific pattern" should also be similarly understood, as long as a certain X-ray powder diffraction pattern can be substantially identified as consistent with the X-ray powder diffraction pattern disclosed in the present application, it should be considered to fall within the protection scope of the present application.

[0086] It can be understood that the differential scanning calorimetry curve and the endothermic peak positions shown therein in the present application should also be similarly understood, and slight discrepancies with the specific values or specific value ranges or specific spectra disclosed in the present application are allowed, but as long as the partial or all endothermic peak positions in the differential scanning calorimetry curve or the entire curve can be substantially identified as consistent with the present application, it should be considered to fall within the protection scope of the present application.

[0087] It is understood that similar interpretations can be made for other spectra that characterize the crystalline type.

[0088] Differential Scanning Calorimetry (DSC)

[0089] Also known as "differential calorimetry scanning analysis", it is a technique that measures the relationship between the energy difference between the measured substance and the reference substance and the temperature during the heating process. The peak position, shape and number of peaks on the DSC spectrum are related to the properties of the substance, so it can be used to qualitatively identify the substance. The method is commonly used in the art to detect various parameters such as phase transition temperature, glass transition temperature, reaction heat, etc. of the substance. The peak position of the DSC spectrum may be slightly different between different instruments, so the numerical value of the peak position of the DSC endothermic peak cannot be considered absolute. According to the condition of the instrument used in the experiment of the present application, the numerical value of the experimental error or difference may be less than or equal to 5°C, or less than or equal to 4°C, or less than or equal to 3°C, or less than or equal to 2°C, or less than or equal to 1°C.

[0090] Identification and properties of crystalline forms

[0091] X-ray powder diffraction

[0092] The compounds of the present application have specific crystalline morphologies, which have specific characteristic peaks in the X-ray powder diffraction pattern (XPRD).

[0093] The XPRD pattern was collected on an X-ray powder diffraction analyzer, and the instrument parameters are shown in Table 1.

[0094] Table 1

[0095]

[0096] In the X-ray powder diffraction pattern, the position of each peak is determined by 2θ (°). It is understood that different instruments and / or conditions can result in slightly different data, and the position and relative intensity of each peak can vary. The intensity of the peak only reflects the approximate size of the peak at each position. In the present application, each crystalline form is defined as the base peak with the highest peak height of the diffraction peak, and its relative intensity is defined as 100%, as I0, and the relative intensity of other peaks is defined as the ratio of the peak height to the base peak height, as I / I0. The definition of the relative intensity of each peak is shown in Table 2.

[0097] Table 2

[0098] Relative intensity II0(%) Definitions 50～100 VS (Very Strong) 25～50 S (Strong) 10～25 M (Medium) 1～10 W (Weak) .

[0099] Sodium valproate and lithium carbonate are commonly used antiepileptic drugs in clinic, and the maintenance dose in treatment is 1000mg-2000mg / day. The excessive dosage often shows related adverse reactions, and the blood lithium concentration needs to be detected during the clinical use of lithium carbonate, and sodium valproate is accompanied by high sodium intake during clinical use. Magnesium valproate can be used to avoid high sodium intake of sodium valproate, however, the effect of magnesium valproate is relatively slow after use, and the action time is relatively long.

[0100] To solve the above problems, the application designs and successfully synthesizes lithium valproate. Lithium valproate avoids high salt intake, and has better effect than lithium carbonate or sodium valproate at the same dose, and has the effect of treating and / or preventing nervous system diseases at a low dose compared with sodium valproate and magnesium valproate.

[0101] The lithium valproate of the embodiment of the application has the following structure:

[0102]

[0103] In some embodiments, the lithium valproate is a lithium salt of valproic acid.

[0104] In some embodiments, the chemical ratio of valproic acid and lithium is 1:1.

[0105] In some embodiments, the lithium salt of valproic acid is a crystalline salt.

[0106] One or more embodiments of the application provide a preparation method of lithium valproate, comprising the following steps:

[0107] Mixing valproic acid, lithium hydroxide and aqueous alcohol solution at 0-10℃ to obtain a first solution;

[0108] Stirring the first solution at 20-40℃ for 12-20h to obtain a second solution;

[0109] Filtering the second solution to collect the solid to obtain lithium valproate;

[0110] The structure of lithium valproate is as follows:

[0111]

[0112] In some embodiments, the volume ratio of alcohol and water in the aqueous alcohol solution is 15-25:1, for example, 15:1, 20:1, 25:1, etc.

[0113] In some embodiments, the alcohol is selected from organic alcohols such as ethanol and methanol.

[0114] In some embodiments, the mass-volume ratio of the aqueous solution of valproic acid and alcohol is 8-12 g: 100 mL, for example 8 g: 100 mL, 9 g: 100 mL, 10 g: 100 mL, 11 g: 100 mL, 12 g: 100 mL, etc., which can ensure that the valproic acid is fully dissolved.

[0115] In some embodiments, the molar ratio of valproic acid and lithium hydroxide is 2:(1-1.5), for example 2:1, 2:1.2, 2:1.5, etc., and a slight excess of lithium valproate can ensure that the reaction is sufficient, and the subsequent valproic acid is easily removed from the system.

[0116] In some embodiments, the valproic acid, lithium hydroxide and alcohol are mixed at 0-10°C to obtain a first solution, which includes:

[0117] After mixing the aqueous solution of valproic acid and alcohol, lithium hydroxide solid is added at 0-10°C, and after the addition is complete, the solution is stirred for a period of time to fully dissolve to obtain a first solution.

[0118] In some embodiments, the first solution is stirred at 20-40°C for 12-20h to obtain a second solution, and the first solution is fully stirred at a suitable temperature to fully react to form lithium valproate.

[0119] In some embodiments, the second solution is filtered to collect the solid to obtain lithium valproate, which includes:

[0120] The second solution is filtered at room temperature, which is usually 20-40°C;

[0121] After filtration, the solution is concentrated at 40-60°C and 10-20 bar;

[0122] After concentration, the solution is vacuum dried at 40-60°C and less than 0.09 MPa to collect the solid to obtain lithium valproate.

[0123] The crystal form of lithium valproate disclosed in the present application has low hygroscopicity, which ensures a lower water content of the product and makes the lithium valproate more stable in quality during storage.

[0124] One or more embodiments of the present application provide a crystal form of lithium valproate, which is selected from any one of the following groups:

[0125] (1) Crystal Form I: its X-ray powder diffraction pattern includes the following characteristic diffraction peaks at 2θ (°) angles: 6.710±0.2°, 7.853±0.2°, 21.674±0.2°, 19.067±0.2°, 20.070±0.2°, 23.617±0.2°, 20.445±0.2° and 15.699±0.2°;

[0126] (2) Form II: an X-ray powder diffraction pattern comprising characteristic diffraction peaks at the following 2-theta (°) angles: 6.660 ± 0.2°, 7.503 ± 0.2°, 18.813 ± 0.2°, 19.522 ± 0.2°, 20.058 ± 0.2°, 22.453 ± 0.2°, 22.729 ± 0.2°, and 15.093 ± 0.2°;

[0127] (3) Form III: an X-ray powder diffraction pattern comprising characteristic diffraction peaks at the following 2-theta (°) angles: 6.703 ± 0.2°, 7.869 ± 0.2°, 21.636 ± 0.2°, 23.632 ± 0.2°, 19.080 ± 0.2°, 20.054 ± 0.2°, 15.713 ± 0.2°, and 27.420 ± 0.2°;

[0128] (4) Form IV: an X-ray powder diffraction pattern comprising characteristic diffraction peaks at the following 2-theta (°) angles: 6.753 ± 0.2°, 7.883 ± 0.2°, 19.169 ± 0.2°, 21.880 ± 0.2°, 20.263 ± 0.2°, 23.716 ± 0.2°, 15.755 ± 0.2°, and 10.930 ± 0.2°;

[0129] (5) Form V: an X-ray powder diffraction pattern comprising characteristic diffraction peaks at the following 2-theta (°) angles: 6.741 ± 0.2°, 7.914 ± 0.2°, 21.680 ± 0.2°, 23.673 ± 0.2°, 19.134 ± 0.2°, 20.094 ± 0.2°, 15.756 ± 0.2°, and 10.847 ± 0.2°;

[0130] (6) Form VI: an X-ray powder diffraction pattern comprising characteristic diffraction peaks at the following 2-theta (°) angles: 6.784 ± 0.2°, 7.946 ± 0.2°, 21.743 ± 0.2°, 19.152 ± 0.2°, 23.707 ± 0.2°, 20.502 ± 0.2°, 20.150 ± 0.2°, and 21.164 ± 0.2°; and,

[0131] (7) Form VII: an X-ray powder diffraction pattern comprising characteristic diffraction peaks at the following 2-theta (°) angles: 6.729 ± 0.2°, 7.702 ± 0.2°, 18.991 ± 0.2°, 20.702 ± 0.2°, 21.741 ± 0.2°, 22.051 ± 0.2°, 22.533 ± 0.2°, and 28.957 ± 0.2°.

[0132] In some embodiments, the crystalline form of lithium valproate has at least one of the following characteristics:

[0133] (1) The X-ray powder diffraction pattern of Form I is substantially as shown in Figure 1

[0134] (2) The X-ray powder diffraction pattern of Form II is substantially as shown in Figure 2

[0135] (3) The X-ray powder diffraction pattern of Form III is substantially as shown in Figure 3

[0136] (4) The X-ray powder diffraction pattern of Form IV is substantially as shown in Figure 4

[0137] (5) The X-ray powder diffraction pattern of Form V is substantially as shown in Figure 5

[0138] (6) The X-ray powder diffraction pattern of Form VI is substantially as shown in Figure 6

[0139] (7) The X-ray powder diffraction pattern of Form VII is substantially as shown in Figure 7

[0140] In some embodiments, the crystalline form of lithium valproate has at least one of the following characteristics:

[0141] (1) The differential scanning calorimetry curve of Form I has an endothermic peak at 359.2±3°C;

[0142] (2) The differential scanning calorimetry curve of Form II has an endothermic peak at 361.8±3°C;

[0143] (3) The differential scanning calorimetry curve of Form III has an endothermic peak at 331.8±3°C;

[0144] (4) The differential scanning calorimetry curve of Form V has an endothermic peak at 357.2±3°C;

[0145] (6) The differential scanning calorimetry curve of Form VI has an endothermic peak at 362.3±3°C; and,

[0146] (7) The differential scanning calorimetry curve of Form VII has an endothermic peak at 362.2±3°C.

[0147] One or more embodiments of the present application provide a method for preparing the above-mentioned crystalline form of lithium valproate, comprising the following steps:

[0148] ​​​​​​​The valproate lithium is crystallized by at least one of the following methods: dissolution crystallization, anti-solvent crystallization, evaporation crystallization, volatile crystallization and melt crystallization; wherein the solvent used includes at least one of the following: ethanol, dichloromethane, acetonitrile, ethyl acetate, tetrahydrofuran, methanol, petroleum ether, butyl acetate, methyl tert-butyl ether, n-butanol, dimethyl sulfoxide, tert-butyl alcohol, isopropanol, toluene and N,N-dimethylformamide; wherein ethyl acetate, dichloromethane and acetonitrile are difficultly soluble solvents, isopropanol, tert-butyl alcohol, butyl acetate, methyl tert-butyl ether, tetrahydrofuran and toluene are slightly soluble solvents, n-butanol is a soluble solvent, and methanol, ethanol and water are easily soluble solvents.

[0149] In this paper, the difficultly soluble solvent refers to that the compound is not dissolved in the solvent by ultrasonic or heating, and a white turbid liquid is obtained; the slightly soluble solvent refers to that the compound is not dissolved in the solvent by heating or ultrasonic, but dispersed into a white dispersion system; the soluble solvent refers to that the compound is dissolved in the solvent by ultrasonic, and a colorless and clear liquid is obtained; and the easily soluble solvent refers to that the compound is uniformly dissolved in the solvent, and a colorless and clear liquid is obtained.

[0150] In this paper, the dissolution crystallization refers to a process that a compound is prepared into a saturated solution with an easily soluble solvent, then mixed with another heated easily soluble solvent, and then the crystals are precipitated; the anti-solvent crystallization refers to a process that a compound is mixed with at least one of a difficultly soluble solvent, a soluble solvent and a slightly soluble solvent, then dissolved by heating, and then the crystals are precipitated by cooling; the evaporation crystallization refers to a process that a compound, an easily soluble solvent and at least one of a difficultly soluble solvent, a soluble solvent and a slightly soluble solvent are mixed and dissolved, and then the crystals are precipitated by concentration; the volatile crystallization refers to a process that a compound is dissolved in an easily soluble solvent, and then the crystals are precipitated by volatilization; and the melt crystallization refers to a process that a compound is heated and melted, and then the crystals are precipitated by cooling; in this application, the temperature of heating and melting is 300 DEG C, and the heating time is 2-24 h.

[0151] One or more embodiments of the present application provide a pharmaceutical composition, which comprises a pharmaceutical active ingredient and at least one of a pharmaceutically acceptable excipient and a carrier;

[0152] The pharmaceutical active ingredient comprises valproate lithium.

[0153] The structure of the valproate lithium is as follows:

[0154]

[0155] As used herein, "pharmaceutically acceptable carrier" shall mean a non-toxic, inert solid, semi-solid or liquid material that is not biologically or otherwise undesirable. The form of the "pharmaceutically acceptable carrier" is not particularly limited and includes, but is not limited to, solid, semi-solid, liquid and the like. The "pharmaceutically acceptable carrier" should be compatible with the patient. The patient is preferably a mammal, more preferably a human. One of the roles of the "pharmaceutically acceptable carrier" is to suitably deliver the active agent to the target site without terminating the activity of the agent. As used herein, the language "pharmaceutically acceptable carrier" includes buffers, sterile water for injection, solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, compatible with pharmaceutical administration. Each carrier must be "pharmaceutically acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient.

[0156] As used herein, "pharmaceutically acceptable excipient" includes, but is not limited to, diluents, wetting agents, binding agents, disintegrating agents, lubricating agents, color, odor, and taste adjusting agents, solvents, solubilizing agents, co-solvents, emulsifiers, antioxidants, metal complexing agents, inert gases, preservatives, local anesthetics, pH adjusting agents, isotonic or isotonizing adjusting agents, and the like. Further: diluents, such as starch, sucrose, cellulose derivatives, inorganic salts, and the like; wetting agents, such as water, ethanol, and the like; binding agents, such as starch paste, dextrin, sugar, cellulose derivatives, gelatin, povidone, polyethylene glycol, and the like; disintegrating agents, such as starch, sodium carboxymethyl starch, low-substituted hydroxypropyl cellulose, sodium carboxymethyl cellulose, cross-linked povidone, surfactants, run-off disintegrating agents, and the like; lubricating agents, such as talc, calcium stearate, magnesium stearate, magnesium lauryl sulfate, microfine silica, polyethylene glycol, and the like; color, odor, and taste adjusting agents, such as pigments, flavorings, sweeteners, gum base, odor correctors, and the like, specifically, fuchsin, xylitol; solvents, such as water, oil, ethanol, glycerol, propylene glycol, polyethylene glycol, dimethyl sulfoxide, liquid paraffin, fatty oil, ethyl acetate, and the like; solubilizing agents, such as Tween, Brij, polyoxyethylene fatty alcohol ether, soap, sulfate, sulfonate, and the like; co-solvents, such as organic acids (e.g., citric acid) and their salts, amide and amine compounds, inorganic salts, polyethylene glycol, povidone, glycerol, and the like; emulsifiers, such as Span, Tween, Brij, benzyl ether, glycerol fatty acid ester, higher fatty acid salt, sulfate, sulfonate, gum arabic, tragacanth gum, gelatin, pectin, phospholipid, agar, sodium alginate, hydroxide, silicon dioxide, bentonite, and the like; suspending agents, such as glycerol, sugar syrup, gum arabic, tragacanth gum, agar, sodium alginate, cellulose derivatives, povidone, carbopol, polyvinyl alcohol, thixotropic agents, and the like; antioxidants, such as sulfite, pyrosulfite, bisulfite, ascorbic acid, gallic acid and its esters, and the like; metal complexing agents, such as disodium ethylenediaminetetraacetate, polycarboxylic acid compounds, and the like; inert gases, such as nitrogen, carbon dioxide, and the like; preservatives, such as nipagin, organic acids and their salts (e.g., sodium benzoate), quaternary ammonium compounds, chlorhexidine acetate, alcohols, phenols, and volatile oils, and the like; local anesthetics, such as benzyl alcohol, trichlorobenzyl alcohol, lidocaine, and procaine, and the like; pH adjusting agents, such as hydrochloric acid, sulfuric acid, phosphoric acid, tartaric acid, acetic acid, sodium hydroxide, sodium bicarbonate, ethylenediamine, meglumine, phosphate, acetate, citric acid, citrate, and the like; isotonic or isotonizing adjusting agents, such as dextrose, sodium chloride, sodium citrate, sorbitol, and xylitol, and the like.It can be understood that the diluent described in the embodiments of the present application can also be called a filler, and plays the same role in the pharmaceutical preparation; the water described in the embodiments of the present application is water meeting the requirements of the preparation, for example, water for injection, purified water and the like, and the oil is oil for injection; the preservative described in the embodiments of the present application can also be called an antibacterial agent, and plays the role of inhibiting the growth of microorganisms, prolonging the shelf life and the like in the preparation; the lubricant in the embodiments of the present application contains a glidant, an anti-adhesive and the like; the sugar described in the embodiments of the present application can be sugar powder or syrup, and the type of the sugar is not limited to glucose; and the flavoring agent includes but is not limited to essence.

[0157] In some embodiments, the pharmaceutical composition comprises a therapeutically effective amount of a pharmaceutically active ingredient.

[0158] As used herein, "therapeutically effective amount" means an amount of a compound of the present application that will elicit the biological or medical response of an individual, for example, reducing or inhibiting the activity of an enzyme or protein or ameliorating symptoms, alleviating the condition, slowing or delaying the progression of a disease, or preventing a disease, and the like.

[0159] One or more embodiments of the present application provide use of the above lithium valproate or pharmaceutical composition in the manufacture of a medicament for treating and / or preventing a nervous system disease.

[0160] As used herein, "preventing and / or treating" means alleviating, slowing progression, attenuating, preventing, or maintaining an existing disease or condition, for example, a nervous system disease. Treatment also includes curing, preventing development of, or alleviating to some extent one or more symptoms of a disease or condition.

[0161] In some embodiments, the nervous system disease comprises at least one of epilepsy, mania, schizofrenia, bipolar disorder, anxiety, depression, and agitation.

[0162] Further, the agitation comprises at least one of schizofrenia agitation, anxiety agitation, depression agitation, and Alzheimer's agitation.

[0163] In some embodiments, the subject of the medicament for treating and / or preventing a nervous system disease is a mammal.

[0164] As used herein, "subject" means an animal, preferably a mammal, and more preferably a human. The term "mammal" refers to a warm-blooded vertebrate class of mammals, including, for example, cats, dogs, rabbits, bears, foxes, wolves, monkeys, deer, mice, pigs, and humans.

[0165] In some embodiments, the mammal comprises at least one of a human and a mouse.

[0166] In some embodiments, the dosage form of the medicament for treating and / or preventing a nervous system disease comprises a solid preparation, a semi-solid preparation, or a liquid preparation. In some embodiments, the dosage form of the medicament for treating and / or preventing a nervous system disease comprises a solid preparation, a semi-solid preparation, or a liquid preparation.

[0167] In some embodiments, the dosage form of the medicament for treating and / or preventing a nervous system disease includes tablets, capsules, granules, powders, dispersants, solutions, suspensions, and the like.

[0168] The mode of administration of the compounds of the present application is not particularly limited.

[0169] Representative modes of administration include, but are not limited to, oral, rectal, parenteral (intravenous, intramuscular, or subcutaneous) injection, and topical administration, inhalation.

[0170] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier) such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, such as starches, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, such as hydroxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and acacia; (c) humectants, such as glycerol; (d) disintegrating agents, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solution retarders, such as paraffin; (f) absorption accelerators, such as quaternary ammonium compounds; (g) wetting agents, such as cetyl alcohol and glycerol monostearate; (h) absorbents, such as kaolin and bentonite clay; and (i) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets, and pills, the dosage forms can also comprise buffering agents. Solid compositions such as tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells, such as enteric coatings and other coatings known in the art. They can contain opacifying agents, and can also be of a composition that they release the active compound or compounds in a certain part of the digestive tract. Examples of embedding compositions that can be used are polymeric substances and waxes. The active compounds can also be in micro-encapsulated form, if desired, with one or more of the excipients described above.

[0171] Liquid dosage forms for oral administration include pharmaceutically- acceptable emulsions, solutions, suspensions, syrups, or elixirs. In addition to the active compounds, the liquid dosage forms can contain inert diluents commonly used in the art, such as water or other solvents, solubilizing agents and emulsifiers, as, for example, ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3- butylene glycol, dimethylformamide, and the like, specifically including mixtures of these substances. Besides such inert diluents, the compositions can also include adjuvants, such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0172] The pharmaceutical compositions can be in unit dosage form. In such form, the composition is divided into unit doses containing appropriate quantities of the active ingredient. The quantity of the active ingredient is predetermined in order to produce the desired therapeutic effect in association with the

[0173] Dosage forms for topical administration of a compound of this application include ointments, powders, sprays, and inhalers. The active component is admixed with a carrier, which can be a solid, a liquid, or a gas, and is applied to the affected area in a manner to be effective in the treatment of the disease or condition.

[0174] It is understood that the pharmaceuticals of the embodiments of the present application can be formulated with different pharmaceutically acceptable excipients to produce suitable clinical dosage forms, including but not limited to the dosage forms described above.

[0175] In some embodiments, the administration frequency is 1-3 times per day, for example, 1 time per day, 2 times per day, 3 times per day.

[0176] In some embodiments, the administration dosage is 250-1000 mg per day.

[0177] In some embodiments, the subject is a mouse, and the administration dosage is 30-200 mg / kg.

[0178] In some embodiments, the subject is a human, and the administration dosage is 4-20 mg / kg.

[0179] In some embodiments, the administration mode is oral.

[0180] In some embodiments, the administration mode is gavage. Gavage in the present application refers to: dissolving the pharmaceutical in a solvent (such as ultrapure water), using a gavage device, and directly injecting the liquid into the stomach of the animal through the mouth of the animal.

[0181] The current clinical mainstream antidepressants are mainly a class of selective serotonin reuptake inhibitors (SSRIs) and serotonin and norepinephrine reuptake inhibitors (SNRIs) and the like. These drugs generally need to start taking effect in 2-4 weeks. Because the effect is slow, for patients with depression, especially severe patients, adverse reactions appear before the effect appears, and patients often do not insist on taking medicine in this 1-2 weeks, and even suicide behavior occurs in this period, which is the clinical pain point of antidepressants. The valproate lithium of the present application has a rapid effect, and can play a therapeutic effect in 8-10 days, effectively improving patient compliance and improving efficacy.

[0182] The valproate lithium of the present application has an improvement effect on the manifestations of various nervous system diseases, including epilepsy, mania, schizophrenia, bipolar disorder, anxiety, depression and agitation, wherein agitation includes schizophrenia agitation, anxiety agitation, depression agitation and Alzheimer's agitation, the mechanism of anti-agitation includes delaying the occurrence of hyperactivity state caused by MK-801 and reducing the degree of hyperactivity caused by MK-801.

[0183] The following are some specific examples.

[0184] The experimental parameters not written in the following specific examples are preferably referred to the guidance given in the present application document, and can also be referred to the experimental manual in the art or other experimental methods known in the art, or the experimental conditions recommended by the manufacturer.

[0185] All or part of the raw materials involved in the following specific examples are referred to Table 1, and the reagents are referred to Table 2, and the remaining raw materials and reagents can be obtained by market or prepared by the skilled person in the art according to known means.

[0186] Table 3

[0187] Material Name Valproic Acid Lithium Hydroxide CAS Number 99-66-1 1310-65-2 Source Shanghai Aldamas Shanghai Aldrin Lot Number P2284353 L128667 Molecular Formula [C8H 16 O2]]> LiOH Molecular Weight 144.21 23.95 Melting Point -21.25 °C (estimate) 470 °C (dec.) (lit.) Boiling Point 220 °C (lit.) 925 °C (lit.) Density 0.9 g / mL at 25 °C (lit.) 1.43 g / cm 3 at 25°C (lit.)

[0188] Table 4

[0189]

[0190] In the following specific examples, the crystal form determination criteria refer to the 2020 edition of "Chinese Pharmacopoeia" 9015 Drug Crystal Form Research and Crystal Form Quality Control Guidelines. If it is determined that the crystal form of two crystal state samples is consistent, the powder X-ray diffraction test should be carried out in parallel, and the diffraction peak number should be the same, the 2θ value diffraction peak position error range of both should be within 0.2°, the relative peak intensity error of the same position diffraction peak should be within ±5%, and the intensity order of the diffraction peak should be consistent. If it is determined that the crystal form of two amorphous state samples is consistent, the geometric topology shape of the diffuse diffraction peak should be completely consistent.

[0191] The experiments and their principles carried out in the following examples are as follows:

[0192] Tail suspension test (TST) is a classic and rapid method for evaluating the efficacy of antidepressants, stimulants, and sedatives. The principle is that after the mouse is suspended, it tries to escape but cannot, and thus gives up struggling and enters a unique state of depression. The immobility time of the animal during the experiment reflects the state of depression, and antidepressants and stimulants can significantly shorten the state.

[0193] Open field test (OFT) is a method for evaluating the behavior of experimental animals, especially in the field of neuropsychiatric research, and is commonly used to measure the spontaneous activity, exploratory behavior, and anxiety and depression-like behavior of animals. Open field test can measure the level of spontaneous activity and exploratory behavior of animals in a new environment, and patients with depression often show reduced activity and lack of interest.

[0194] Sucrose preference test is a classic experiment for detecting the symptom of pleasure deficiency in depression. It is a detection method designed by using the preference of rodents for sweet taste. After the animals are fasted for a period of time, white water and low concentration sucrose water are given at the same time, and the preference degree of animals for sucrose water (sucrose preference degree) is used as an index to detect whether the animals have the symptom of pleasure deficiency.

[0195] The mice used in the following examples are C57BL / 6j mice, which are commonly used animals for pharmacological, pharmacodynamic and disease model exploration of test products for or intended for human use. In terms of gender, considering that the changes in sex hormones in female mice can have a greater impact on behavior and neurochemical indicators, it is recommended to use male C57BL / 6j mice.

[0196] The term (+)-MK-801 is a highly selective, strong inhibitory non-competitive NMDA receptor antagonist, which can mimic the typical positive symptoms, negative symptoms and cognitive impairment symptoms of schizophrenia, and is considered one of the best drugs for modeling schizophrenia.

[0197] Example 1 Preparation of lithium valproate

[0198] 1.1 Synthesis route

[0199]

[0200] 1.2 Preparation method

[0201] (1) Prepare a 1 L three-necked flask, add valproic acid (28.8 g, 2 mol) and ethanol / purified water (273.6 mL / 14.4 mL), reduce the system to 0-10 °C, add lithium hydroxide solid (5.7 g, 2.4 mol, 1.2 eq.) to the system, mix well, and stir at room temperature 20-30 °C overnight for about 18 hours;

[0202] (2) Filter at room temperature, concentrate the filtrate at 50 °C and 14 bar, and vacuum dry at 50 °C and less than 0.09 MPa to obtain lithium valproate powder.

[0203] 1.3 Characterization results

[0204] HRMS data: [M+H] + Calculated value = 151.1305; [M+H] + Actual value = 151.1306;

[0205] Hydrogen spectrum data: 1 H NMR (CD3OD, 400 MHz): δ 2.19 (1H, m), 1.59-1.48 (2H, m), 1.42-1.25 (6H, m), 0.90 (6H, t, J = 7.2 Hz);

[0206] Carbon spectrum data: 13 C NMR (CD3OD, 101 MHz): δ 185.3, 50.2, 36.9, 22.0, 14.5.

[0207] Example 2 Preparation of lithium valproate crystal form

[0208] 2.1 Crystal form I

[0209] (1) Take lithium valproate crude product (1.0 g, 6.7 mmol) in a round-bottom flask of appropriate size, and add tetrahydrofuran (30 mL), and react at reflux temperature overnight.

[0210] (2) Filter while hot, and rinse the filter cake with tetrahydrofuran (10 mL), and vacuum dry at 100 °C and less than 0.09 MPa for 18 hours.

[0211] 2.2 Crystal form II

[0212] (1) Take a round-bottom flask of appropriate size, add acetonitrile (100 mL), and warm to 60 °C; add a saturated aqueous solution (10 mL) of lithium valproate (3.6 g, 24 mmol) dropwise into the above hot acetonitrile, stir for 20 min after the dropwise addition is complete, turn off the heating, and stir at room temperature overnight.

[0213] (2) Filter at room temperature, wash the filter cake with a small amount of acetonitrile, and dry under vacuum at 50°C and less than 0.09 MPa for 7 hours.

[0214] 2.3 Form III

[0215] (1) Take a round bottom flask of appropriate size, add solvent II (100 mL / 150 mL, 100 V / 150 V), and warm to 60°C. Add a saturated solution I (2.8 V / 10 V) of crude valproate lithium (3.6 g / 3.0 g / 1.5 g, 24 mmol / 20 mmol / 10 mmol, 1.0 equivalent) in solvent II dropwise. After the dropwise addition is complete, stir for 20 min, turn off the heat, and stir overnight at room temperature.

[0216] (2) Filter at room temperature, wash the filter cake with a small amount of solvent II, and dry under vacuum at 50°C / <0.09 MPa for 7 hours. Take a sample for testing.

[0217] 2.4 Form IV

[0218] (1) Take crude valproate lithium (1.0 g, 6.7 mmol) in a round bottom flask of appropriate size, and add ethyl acetate (100 mL) and ethanol (20 mL). Dissolve under ultrasonic conditions, filter at room temperature, and concentrate to a total solvent volume of 24 mL-36 mL at an appropriate temperature. Crystallize at room temperature overnight.

[0219] (2) Filter at room temperature, and rinse the filter cake with ethyl acetate (5 mL). Dry under vacuum at 100°C and less than 0.09 MPa for 18 hours.

[0220] 2.5 Form V

[0221] (1) Prepare a crude valproate lithium solution: Take a 40 mL sample bottle, and add 200 mg of crude valproate lithium to each sample bottle. Dissolve under ultrasonic conditions with 2 mL of methanol to obtain a clear 100 mg / mL crude valproate lithium solution in methanol, which is ready for use.

[0222] (2) Take 1.5 mL of an ep tube, and add a total of 15-25 mg of crude valproate lithium solution to each ep tube. Add an equal volume of petroleum ether, and dissolve the partially crystallized system under ultrasonic conditions. Leave the ep tube cap slightly open, and store it at room temperature in the dark. Observe the solvent evaporation every day and record it until the solvent is completely evaporated. Dry under vacuum at 100°C and less than 0.09 MPa.

[0223] 2.6 Form VI

[0224] Place crude valproate lithium in a muffle furnace, and roast at 300°C for 2 hours. Cool down.

[0225] 2.7 Crystal Form VII

[0226] (1) Take a round bottom flask of appropriate size, add tetrahydrofuran (100 mL), and warm to 60 °C. Add a saturated aqueous solution (10 mL) of crude lithium valproate (3.6 g, 24 mmol) in tetrahydrofuran dropwise. After the addition is complete, stir for 20 min, turn off the heat, and stir overnight at room temperature.

[0227] (2) Filter at room temperature, wash the filter cake with a small amount of tetrahydrofuran, and vacuum dry at 50 °C / <0.09 MPa for 7 h. Take a sample for testing.

[0228] 2.8 Crystal Form Summary

[0229] Table 5 Crystal Form Summary

[0230]

[0231] Example 3 Investigation of the Hygroscopicity of Different Crystal Forms of Lithium Valproate

[0232] 3.1 Reagents and Equipment

[0233] The reagent information is shown in Table 6, and the equipment information is shown in Table 7.

[0234] Table 6

[0235] Sample Name Sample Source Lithium Valproate Form I Form I of Example 2 Lithium Valproate Form III Form III of Example 2 Lithium Valproate Form IV Form IV of Example 2 Lithium Valproate Form V Form V of Example 2 Lithium Valproate Form VI Form VI of Example 2 Sodium Valproate Purchased

[0236] In Table 6, the hygroscopicity curve of crystal form VI was investigated after melting at 300 °C in a muffle furnace for 2 h.

[0237] Table 7

[0238]

[0239] 3.2 Investigation Method

[0240] The hygroscopicity of the reference drug was determined according to the Guidance for the Hygroscopicity Test of Drugs (Chinese Pharmacopoeia 2020 Edition Volume IV General Rules 9103), including the following steps:

[0241] (1) Take a dry stoppered glass weighing bottle, and pre-saturate it in a suitable 25 °C ± 1 °C constant temperature dryer (select the saturated aqueous solution according to different humidity) or artificial climate chamber (temperature 25 °C ± 1 °C, set a series of humidity) the day before the test. Precisely weigh the weight (m1).

[0242] (2) Precisely take an appropriate amount of test sample, and spread it in the above weighing bottle. The thickness of the test sample is about 1 mm, and precisely weigh (m2).

[0243] (3) Place the weighing bottle with the cap in the above constant temperature and humidity conditions for 24 h. Precisely weigh (m3).

[0244] (4) Calculation formula:

[0245] Weight gain percentage = m 3 -m 2 × 100%

[0246] m2-m1

[0247] (5) Hygroscopicity characteristic description and definition of hygroscopic weight gain:

[0248] Deliquescence: Absorbing sufficient moisture to form a liquid;

[0249] Extremely hygroscopic: Hygroscopic weight gain is not less than 15%;

[0250] Hygroscopic: Hygroscopic weight gain is less than 15% but not less than 2%;

[0251] Slightly hygroscopic: Hygroscopic weight gain is less than 2% but not less than 0.2%;

[0252] Non-hygroscopic or almost non-hygroscopic: Hygroscopic weight gain is less than 0.2%.

[0253] 3.3 Investigation results

[0254] Table 8 Hygroscopicity curve investigation results

[0255]

[0256] According to the statistical data in Table 8, it can be found that the hygroscopicity of each crystal form of lithium valproate is weaker than that of sodium valproate, and is more stable. Lithium valproate crystal forms I, III, IV and V have hygroscopicity, and crystal form VI has no hygroscopicity. Crystal form VI is a stable crystal form.

[0257] Example 4 Application of lithium valproate in the preparation of an antiepileptic drug

[0258] This example provides the application of the lithium valproate of the present application in the preparation of an antiepileptic drug, which is verified by exploratory tests on an acute pentylenetetrazole epilepsy ignition model. The reagents used in this example are prepared in a sterile environment.

[0259] 4.1 Experimental materials

[0260] (1) Reagents

[0261] The lithium valproate prepared in Example 1 was subjected to exploratory tests on an acute pentylenetetrazole epilepsy ignition model.

[0262] Ultra-pure water was used as the solvent for preparation. Taking the preparation of 2 mL of the test sample as an example, the preparation parameters are shown in Table 9.

[0263] Table 9

[0264]

[0265] Positive control 1: sodium valproate (Shanghai Aladdin Bio-Chem Technology Co., Ltd., white solid powder, 500 g / bottle)

[0266] Preparation of positive control 1: ultrapure water was used as the solvent for preparation. For example, 2 mL of the test sample was prepared, and the preparation parameters are shown in Table 10.

[0267] Table 10

[0268]

[0269] Positive control 2: lithium carbonate (Shanghai Jizhi Biochemical Technology Co., Ltd.; white solid powder; 25 g / bottle)

[0270] Preparation of positive control 2: ultrapure water was used as the solvent for preparation. For example, 2 mL of the test sample was prepared, and the preparation parameters are shown in Table 11.

[0271] Table 11

[0272]

[0273] (2) Reagents

[0274] Modeling reagent: pentylenetetrazole (Sigma Aldrich (Shanghai) Trading Co., Ltd., white powder)

[0275] Preparation of modeling reagent: physiological saline was used for preparation. For example, 15 mL of solution was prepared, and the preparation parameters are shown in Table 12.

[0276] Table 12

[0277]

[0278] Other main reagents:

[0279] Sodium chloride injection (Shandong Qidu Pharmaceutical Co., Ltd., 0.9 g / 100 mL, batch number G124040605)

[0280] (3) Equipment

[0281] Electronic analytical balance: manufacturer METTLLER TOLEDO, model ML2001.

[0282] 4.2 Selection of experimental animals

[0283] SPF level C57BL / 6j mice were selected, and the provider was Chengdu Yakang Biological Technology Co., Ltd.

[0284] 4.3 Animal modeling and grouping

[0285] Modeling time: 1 hour after administration of the test sample;

[0286] Modeling animals: all groups of animals;

[0287] Modeling method: 1 hour after intragastric administration of the test product, the animals were injected intraperitoneally with pentylenetetrazol (60 mg / kg, 10 mL / kg).

[0288] Group design: model control group, sodium valproate group, lithium carbonate group, lithium valproate group;

[0289] Number of animals: a total of 48, 12 in each group;

[0290] Gender ratio: all male;

[0291] Grouping method: according to the body weight of the mice, random numbers were assigned, and the random numbers were sorted in descending order. The random numbers were sorted from low to high as the 1st, 2nd, 3rd, and 4th groups, respectively, representing the model control group, sodium valproate group, lithium carbonate group, and lithium valproate group. The specific grouping information is shown in Table 13.

[0292] Table 13

[0293]

[0294] In Table 13, the first digit of the animal number represents the group (1, 2, 3, 4, representing the model control group, sodium valproate group, lithium carbonate group, and lithium valproate group, respectively). The second letter represents the gender (M for male), and the last three digits represent the animal serial number. The "-" represents not applicable.

[0295] The dosages, concentrations, and volumes of administration are shown in Table 13.

[0296] Frequency and cycle of administration: once a day;

[0297] The day of administration is defined as the first day of the test (day 1, D1).

[0298] 4.4 Monitoring of animal status

[0299] (1) General state observation

[0300] Observation time: once a day;

[0301] Observed animals: all active animals in the group;

[0302] Observation content: the main observation content includes but is not limited to the administration site (whether there are symptoms such as redness, bleeding, etc.), behavior state, eyes, mouth, nose, ears, hair, and symptoms such as feces, urine, and genital organs.

[0303] (2) Body weight

[0304] Measurement time: weigh at least twice during the adaptation period, and weigh once before administration;

[0305] Animals for determination: all animals in adaptation period, all animals in test period.

[0306] (3) Epilepsy grade score

[0307] Animals for determination: all animals in each group;

[0308] Time for determination: immediately after modeling;

[0309] Method for determination: immediately after the injection of pentylenetetrazol, the mouse was put into a transparent glass cylinder, and the behavior of the animal was observed and recorded for 30 min, and the video was recorded. The latency of seizure, the number of seizures in 30 min, the total duration of seizures in 30 min, the highest Racine grade of seizure, and the evaluation criteria of Racine epilepsy grade are shown in Table 14.

[0310] Table 14

[0311]

[0312] 4.5 Experimental results

[0313] (1) General state observation

[0314] During the test period, no obvious abnormalities were observed in all animals.

[0315] (2) Body weight

[0316] The body weight data of mice is shown in Table 15. The body weight of mice in each group increased steadily throughout the test period, and the growth trend was consistent, with no significant difference.

[0317] Table 15 Animal body weight data

[0318]

[0319] The data in Table 15 are expressed as mean (Mean) ± standard deviation (SD).

[0320] (3) Seizure

[0321] A large dose of pentyl nitrate single administration can induce acute seizures in animals. In this experiment, all mice developed grade 5 seizures after injection of pentyl nitrate, with a seizure rate of 100%, so the effect of each drug on epilepsy can be reflected by the latency of grade 5 seizures in each group of mice. The latency of grade 5 seizures in mice is shown in Table 16. According to Table 16, after injection of pentyl nitrate, the latency of grade 5 seizures in each group of mice was 123.0 ± 25.5 s for the model control group, 178.6 ± 73.0 s for the sodium valproate group, 142.5 ± 40.7 s for the lithium carbonate group, and 255.6 ± 190.3 s for the lithium valproate group. Among them, sodium valproate can prolong the average latency of grade 5 seizures in mice by about 50 s, lithium carbonate can prolong the average latency of grade 5 seizures in mice by about 20 s, and lithium valproate can prolong the average latency of grade 5 seizures in mice by about 130 s.

[0322] The above results show that lithium valproate can significantly delay grade 5 seizures caused by pentyl nitrate, and at the same dose, the effect is better than that of sodium valproate and lithium carbonate.

[0323] Table 16 Latency of seizures

[0324]

[0325] The data in Table 16 are expressed as mean ± standard deviation (SD).

[0326] Example 5 Study on the rapid antidepressant effect of lithium valproate

[0327] 5.1 Selection of experimental animals

[0328] SPF C57BL / 6J mice (male, 7 weeks) 42 (manufacturer: Zhejiang Weitong Lihua Experimental Animal Technology Co., Ltd., license number: SCXK (Su) 2023-0009), feeding conditions (ordinary feed, temperature 20-26℃, humidity 40%-70%)

[0329] During transportation, feeding, modeling and killing, the interests of animals are fully considered, animals are treated well, stress, pain and harm to animals are prevented or reduced, animal life is respected, and cruel behavior against animals is prohibited, and the least painful method is used to dispose of animals. The method and purpose of animal experiment comply with the moral and ethical standards of human beings and international practices.

[0330] 5.2 Experimental materials

[0331] (1) Experimental reagents and instruments refer to Tables 17-18

[0332] Table 17 Reagent information

[0333]

[0334] Table 18 Main instrument information

[0335]

[0336] (2) Reagent preparation method

[0337] Preparation of 0.1 mg / ml corticosterone drinking water: 420 mg of corticosterone was weighed into 42 ml of absolute ethanol, and stirred magnetically at 1000 r / min until dissolved. 4158 ml of purified water was then added, and the solution was used in the dark.

[0338] Preparation of fluoxetine (10 mg / kg, 10 ml / kg, 1 mg / ml): 125 mg of CMC-Na was weighed into 25 ml of purified water, and stirred magnetically until dissolved. 25 mg of fluoxetine was then added, and stirred magnetically until dissolved (2000 r / min).

[0339] Low, medium and high dose groups were prepared using the lithium valproate powder prepared in Example 1.

[0340] Preparation of low dose (45.5 mg / kg, 10 ml / kg, 4.55 mg / ml): 53.9 mg of lithium valproate was weighed into 11.84 ml of purified water, and dissolved thoroughly.

[0341] Preparation of medium dose (90.1 mg / kg, 10 ml / kg, 9.01 mg / ml): 98.6 mg of lithium valproate was weighed into 10.94 ml of purified water, and dissolved thoroughly.

[0342] Preparation of high dose (135.2 mg / kg, 10 ml / kg, 13.52 mg / ml): 142.4 mg of lithium valproate was weighed into 10.53 ml of purified water, and dissolved thoroughly.

[0343] 5.3 Animal modeling and grouping

[0344] (1) Animal grouping

[0345] Normal control group (no intervention treatment) (N = 8)

[0346] Model + vehicle group (vehicle treatment, gavage administration, 1 time / animal / day, continuous treatment for 12 days) (N = 6)

[0347] Model + positive drug group (fluoxetine treatment, dose: 10 mg / kg, gavage administration, 1 time / animal / day, continuous treatment for 12 days) (N = 7)

[0348] Model + low dose - test drug group (test compound treatment, dose: 45.5 mg / kg, gavage administration, 1 time / animal / day, continuous treatment for 12 days) (N = 7)

[0349] Model group + high dose - test drug group (treatment of test compound, dose: 135.2 mg / kg, gavage, 1 time / each / day, continuous treatment for 12 days) (N = 7)

[0350] Model group + high dose - test drug group (treatment of test compound, dose: 135.2 mg / kg, gavage, 1 time / each / day, continuous treatment for 12 days) (N = 7)

[0351] (2) Animal modeling

[0352] After adaptive feeding, all animals were weighed, the normal control group was not stimulated and was given normal drinking water, all mice except the normal control group were single-caged, and 0.1 mg / ml corticosterone drinking water and chronic unpredictable mild stimulation were given, and the specific time arrangement is shown in Tables 3 and 4. Model verification was started on the 27th day of modeling, including tail suspension test, forced swimming test and open field test.

[0353] 5.4 Animal experiment

[0354] After successful modeling, grouping and treatment were performed according to 5.3, except that the blank control group was not subjected to any intervention treatment, and the rest of the groups were given corresponding drugs (solvent, positive drug, low, medium and high dose - test drug) treatment, gavage, 1 time / each / day, continuous treatment for 12 days, and the modeling intervention was maintained during the treatment period. The behavior experiment including tail suspension test, open field test, sugar water preference test and forced swimming test was performed 24 hours after the 10th treatment.

[0355] (1) Tail suspension test

[0356] The tail suspension test was performed on modeling D27 and treatment D9. A 3 cm long wooden stick was bound at the root of the tail to avoid the animal climbing the tail, and the animal tail tip was fixed so that the mouse was in an inverted state for 6 min. The rest time of each mouse in the last 4 min was recorded. The rest time refers to the duration of the animal stopping struggling and the limbs being motionless.

[0357] (2) Forced swimming

[0358] The forced swimming test was performed on modeling D28 and treatment D15. The animal was placed alone in a plastic basket (30 cm long, 20 cm wide, 15 cm high, water depth 10 cm) containing water, and forced to swim for 6 min. The KEMaze animal behavior video analysis software system was used to automatically record and analyze the activity state of the mouse within 6 min, and the cumulative immobility time of the animal was counted (immobility refers to the mouse stopping struggling or the mouse being in a floating state, and the mouse has slight movement to keep the head above the water surface). Note that the water is changed after each animal test to remove the influence of other mouse odors.

[0359] (3) Open field test

[0360] The open field test was performed on the day of modeling D30 and the day of treatment D10. The animals were placed in the central area of the open field box, and after the animals were allowed to freely move in the test box for 1 min of adaptation, the KEMaze animal behavior video analysis software system was used to automatically record and analyze the activity state and movement trajectory of the mice within 5 min. The open field box was cleaned before each experiment to prevent the information (such as the size of the animal's urine and odor) left by the animal from affecting the test results of the next time.

[0361] (4) Sugar water preference test

[0362] The sugar water preference test was performed from D10 to D13 of the treatment. On D10, all animals began sugar water preference training, and all animals were given 2 bottles of 3% sugar water (100 ml / bottle). On D11, all animals were given 1 bottle of 3% sugar water and 1 bottle of pure water (100 ml / bottle), and the positions of the bottles were changed at about 11 o'clock, 14 o'clock and 18 o'clock. On D12, all animals began to fast and abstain from water for 24 hours. After the fast and water abstinence ended on D13, the formal sugar water preference test began, and all animals were given 1 bottle of 3% sugar water and 1 bottle of pure water (100 ml / bottle). The initial weights of the two bottles were recorded, and the positions of the bottles were changed at 17:30, the next day at 9 o'clock and the next day at 12 o'clock. After 24 hours, the sugar water test ended, and the remaining weights of the two bottles were weighed and recorded. The sugar water preference index was calculated.

[0363] 5.5 Experimental results

[0364] 5.5.1 Evaluation of the mouse depression model

[0365] (1) Changes in body weight during modeling

[0366] Figure 8 The graph of the changes in body weight of the mice during the modeling of the mouse depression model of Example 5. According to the results, the body weight of the model group was significantly lower than that of the normal group. Figure 8 It can be found that the body weight of the model group was significantly lower than that of the normal group.

[0367] (2) Tail suspension behavior test of depressive mice

[0368] Figure 9 The graph of the stillness time of the tail suspension test of the mouse depression model of Example 5 (***p<0.001, compared with the normal group). In the tail suspension test, the total stillness time of the model group was significantly higher than that of the normal group (P<0.001).

[0369] (3) Open field behavior test of depressive mice

[0370] Figure 10The statistical chart of the open field test data of the depression model mice of Example 5, in the open field test, the total distance and average speed of the model group were significantly reduced (P<0.001), and the immobility time was significantly increased (P<0.001).

[0371] According to the results of the above weight, tail suspension test, and open field test, it is shown that the modeling is successful.

[0372] 5.6.2 Pharmacodynamic evaluation of the test drug

[0373] (1) Effect of the test drug on the sucrose preference behavior of the depression mice

[0374] Figure 11 The statistical chart of the sucrose preference rate of the depression model mice of Example 5 affected by the test drug, in the sucrose preference test, compared with the control group, the sucrose preference rate of the model group mice was significantly decreased (P<0.01), with a decrease rate of 15.6%, and the hedonic deficit was reduced, the positive drug and the low-dose test drug group had no obvious effect on the sucrose preference, and the medium- and high-dose test drug groups significantly increased the sucrose preference after treatment, the medium-dose group increased the sucrose preference degree by 8.2%, and the high-dose group increased by 10.1%, indicating that the medium- and high-dose groups increased the hedonic experience and reduced the depression symptoms.

[0375] (2) Effect of the test drug on the tail suspension test behavior of the depression mice

[0376] Figure 12 The statistical chart of the tail suspension test immobility time of the depression model mice of Example 5 affected by the test drug, in the tail suspension test, the total immobility time of the model group was higher than that of the normal group, increased by 43.4%, and due to the abnormality of individual mice, the difference within the group was not significant, the average immobility time of the positive drug group and the high-dose test drug group was lower than that of the model group, the immobility time of the positive drug was reduced by 9.2%, and the immobility time of the high-dose test drug group was reduced by 18.5%, indicating that the test drug can shorten the "immobile state" and the effect is stronger than that of the positive drug group.

[0377] (3) Effect of the test drug on the forced swimming behavior of the depression mice

[0378] Figure 13 The results of the forced swimming behavior of the depression model mice of Example 5 affected by the test drug, in the forced swimming test, compared with the model group, the immobility time of the positive drug group was significantly decreased, with a decrease rate of 9.5%, and the immobility time of the low-, medium-, and high-dose groups was also significantly decreased, with a decrease rate of 4.2%, 11.2%, and 15.8%, respectively, indicating that the treatment effect of the medium- and high-dose groups will be better than that of the positive drug group.

[0379] Figure 14The results of the open field behavior of the mice in the depression model of Example 5 affected by the test drug were obtained. In the open field test, the total distance and average speed of the model group were significantly reduced (P<0.001), and the immobility time was significantly increased (P<0.001), while the positive drug and the test drug had no significant effect on the depression mice after treatment. The open field test is an important experiment for evaluating anxiety and exploratory behavior, while the efficacy of the positive drug and the test drug is mainly reflected in the aspect of anti-depression, and the doses of anxiety and depression are different, and the dose used in this experiment is calculated according to the clinical dose for the treatment of depression, so there is no significant effect on the indicators of the open field test.

[0380] From the behavioral results after treatment, it can be seen that the test drug has an improving effect on the depression behavior of mice, reduces the immobility time of tail suspension and forced swimming, and improves the sucrose preference rate, and the effect of the high-dose test drug is obviously better than that of the positive drug.

[0381] Based on the experimental results, the lithium valproate of the present application has a better anti-depression effect than the positive drug, and the lithium valproate of the present application also has a rapid anti-depression effect.

[0382] Example 6 Lithium valproate for MK801-induced anti-excitation experiment

[0383] 6.1 Experimental materials

[0384] (1) Reagents

[0385] The lithium valproate prepared in Example 1 was subjected to the MK801-induced anti-excitation experiment.

[0386] Ultra-pure water was used as the solvent for preparation. Taking the preparation of 2 mL of the test sample as an example, the preparation parameters are shown in Table 19.

[0387] Table 19

[0388]

[0389] (2) Reagents

[0390] Modeling reagent: (+)-MK-801 hydrogen maleate (MK801) (Sigma Aldrich (Shanghai) Trading Co., Ltd., white powder)

[0391] Preparation of modeling reagent: physiological saline was used for preparation. Taking the preparation of 10 mL of solution as an example, the preparation parameters are shown in Table 20.

[0392] Table 20

[0393]

[0394] Other main reagents:

[0395] Sodium chloride injection (Shandong Qidu Pharmaceutical Co., Ltd., 0.9 g / 100 mL, batch number G124040605)

[0396] (3) Equipment

[0397] The equipment information is shown in Table 21.

[0398] Table 21

[0399]

[0400] 6.2 Selection of experimental animals

[0401] SPF grade C57BL / 6j mice were selected, and the provider was Chengdu Yake Biological Technology Co., Ltd.

[0402] 6.3 Animal modeling and grouping

[0403] Modeling time: 1 hour after administration of the test product;

[0404] Modeling mice: model group, low-dose group, medium-dose group, and high-dose group;

[0405] Modeling method: every 4 mice were a batch (one mouse in the model group, low-dose group, medium-dose group, and high-dose group), 1 hour after intragastric administration of the test product, MK-801 (0.3 mg / kg, 10 mL / kg) was injected intraperitoneally, and then the mice were directly placed in the open field for open field detection, video recording, and observation of the spontaneous activity of the mice within 2 hours. After all the modeling mice were detected, the normal control group mice were detected.

[0406] Group design: normal control group, model control group, low-dose group, medium-dose group, and high-dose group;

[0407] Number of animals: a total of 30, 6 in each group;

[0408] Gender ratio: all male;

[0409] Grouping method: according to the body weight of the mice, random numbers were assigned, and the random numbers were sorted in descending order, and the random numbers were sorted from low to high as the 1st, 2nd, 3rd, 4th, and 5th groups, respectively, representing the normal control group, the model control group, and the three test product dose groups. The specific grouping information is shown in Table 22.

[0410] Table 22

[0411]

[0412] The first digit of the animal number in Table 22 represents the group (1, 2, 3, 4, 5 represent normal control group, model control group, low-dose group, medium-dose group, high-dose group, respectively). The second letter represents gender (M for male), and the last three digits represent the animal serial number. The "-" represents not applicable.

[0413] The administration dose, concentration, and volume are shown in Table 23.

[0414] The administration frequency and cycle are once a day.

[0415] The administration day is defined as the first day of the test (day 1, D1).

[0416] 6.4 Animal state monitoring

[0417] (1) General state observation

[0418] Observation time: once a day;

[0419] Observation animals: all active animals in the group;

[0420] Observation content: the main observation content includes but is not limited to the administration site (whether there are symptoms such as redness, bleeding, etc.), behavior state, eyes, mouth, nose, ears, hair, and symptoms such as feces, urine, and genital organs. If abnormalities occur, detailed description is required.

[0421] (2) Body weight

[0422] Measurement time: at least twice during the adaptation period, and once before administration;

[0423] Measurement animals: all animals during the adaptation period, and all animals in the group during the test period.

[0424] (3) Open field test

[0425] Detection animals: all animals in the group;

[0426] Detection time: immediately after modeling;

[0427] Detection method: the experiment is carried out in a quiet environment. Mice move in the open field box, the bottom is divided into central and peripheral regions, and a camera is installed at the top of the center, connected to a recording system. At the beginning of the experiment, the mice are placed in the central region, and the movement trajectory and spontaneous activity of the mice are recorded for 2 hours. After the experiment of a batch of mice is recorded, the feces and urine are cleaned, and the plastic box bottom and surrounding area are thoroughly cleaned with new jie'er, and then the next batch of animal experiments are carried out after ventilation and drying to prevent odor interference.

[0428] 6.5 Experimental results

[0429] (1) General state observation

[0430] During the experiment, none of the animals showed any obvious abnormalities.

[0431] (2)Weight

[0432] The weight data of the mice are shown in Table 25. Throughout the experimental period, the mice in each group showed a stable increase with a consistent growth trend and no significant differences. The results indicate that the administration of lithium valproate by gavage had no significant effect on the weight of the mice.

[0433] Table 23 Animal weight data

[0434]

[0435] The data in Table 23 are expressed as mean ± standard deviation (SD).

[0436] (3) Spontaneous activities in open spaces

[0437] Following a single dose of MK-801, animals exhibited hyperactivity, mimicking positive symptoms of agitation in schizophrenia. The effect lasted for approximately 2 hours. Therefore, open field analysis was used to measure the distance mice traveled within 2 hours after MK-801 injection to reflect the degree of hyperactivity. The mouse movement distance is shown below. Figure 15 , Figure 15 The above are the statistical results of the effect of lithium valproate on the movement distance of model animals in Example 5. Figure 15 In the graph, A represents the distance moved by mice in each group within 10 minutes after injection of MK-801 during the experiment. A total of 120 minutes were recorded, which is a line graph showing the movement distance of mice within 12 10-minute intervals. Figure 15 B represents the total distance traveled by mice in each group within 120 minutes. Figure 15 All data are expressed as mean ± standard error (SEM).

[0438] according to Figure 15The model group mice showed hyperactivity within 10 minutes after injection of MK-801, and the movement distance was higher than that of the normal control group. The model group mice reached the peak of hyperactivity at about 50-80 minutes after injection. After 80 minutes, the effect of MK-801 gradually decreased, and the movement distance of the model group mice also gradually decreased. The total movement distance of the model group mice within two hours after modeling was significantly increased compared with that of the normal control mice. The lithium valproate groups did not show hyperactivity within 20 minutes after injection of MK-801, and the movement distance was close to that of the normal control group. The lithium valproate groups gradually showed hyperactivity 20 minutes after injection of MK-801, and the movement distance within each 10 minutes was lower than that of the model group. After 80 minutes of injection of MK-801, the movement distance of the lithium valproate groups gradually approached and finally leveled off with that of the model group, and the movement distance within each 10 minutes of the low-dose lithium valproate group was lower than that of the model group. The total movement distance of the lithium valproate groups within two hours after modeling was reduced to some extent compared with that of the model control group, but the total movement distances of the lithium valproate groups were close to each other.

[0439] The above results show that lithium valproate can delay the occurrence of hyperactivity caused by MK-801 to some extent, reduce the degree of hyperactivity by about 20%, and the effect has no obvious dose dependence.

[0440] All the documents mentioned in the present application are incorporated by reference in the present application as if each document is individually incorporated by reference. Unless and to the extent that the documents mentioned in the present application conflict with the application purpose and / or technical scheme of the present application, the documents mentioned in the present application are incorporated by reference in their entirety and in their entirety purpose. When the present application refers to the documents mentioned in the present application, the definitions of the relevant technical features, terms, nouns, phrases, etc. in the documents are also incorporated by reference. When the present application refers to the documents mentioned in the present application, the examples and preferred modes of the relevant technical features are also incorporated by reference into the present application, but are limited to the implementation of the present application. It should be understood that when the content of the reference conflicts with the description in the present application, the present application is used as the reference or is modified according to the description in the present application.

[0441] The technical features of the above embodiments and examples can be combined in any suitable manner. In order to make the description concise, not all possible combinations of the technical features in the above embodiments and examples are described, but as long as the combinations of the technical features do not conflict, they should be considered within the scope of the present description.

[0442] The above examples only express several embodiments of the present application, but cannot be interpreted as a limitation to the patent scope. It should be pointed out that, for those skilled in the art, several modifications and improvements can be made without departing from the concept of the present application, and these all belong to the protection scope of the present application. In addition, it should be understood that, after reading the above teaching of the present application, those skilled in the art can make various changes or modifications to the present application, and the equivalent forms thus obtained also fall within the protection scope of the present application. It should also be understood that, on the basis of the technical solutions provided by the present application, those skilled in the art can obtain technical solutions through logical analysis, reasoning or limited experiments, and these all fall within the protection scope of the appended claims of the present application. Therefore, the protection scope of the patent of the present application should be subject to the appended claims, and the description can be used to explain the content of the claims.

Claims

1. A lithium valproate, having the following structure: ； The lithium valproate is a lithium salt of valproic acid, wherein the chemical ratio of valproic acid to lithium is 1:

1. Its features are, The lithium salt of the valproic acid is a crystalline salt; The crystal form of the crystalline salt is crystal form VI, and its X-ray powder diffraction pattern includes the following characteristic diffraction peaks at 2θ (°): 6.784±0.2°, 7.946±0.2°, 21.743±0.2°, 19.152±0.2°, 23.707±0.2°, 20.502±0.2°, 20.150±0.2°, and 21.164±0.2°.

2. The lithium valproate as described in claim 1, characterized in that, The differential scanning calorimetry curve of crystal form VI has an endothermic peak at 362.3±3℃.

3. A method for preparing lithium valproate according to claim 1 or 2, characterized in that, Includes the following steps: The first solution is obtained by mixing valproic acid, lithium hydroxide and an aqueous solution of alcohol at 0℃~10℃; The first solution is stirred at 20℃~40℃ for 12h~20h to obtain the second solution; The second solution was filtered, and then concentrated at 40–60°C and 10–20 bar. The solid was collected to obtain the lithium valproate.

4. A method for preparing lithium valproate according to claim 1 or 2, characterized in that, Includes the following steps: Lithium valproate was taken and crystallized by melt crystallization.

5. A pharmaceutical composition, characterized in that, The pharmaceutical composition includes at least one of a pharmaceutically acceptable excipient and a carrier; The active pharmaceutical ingredient includes lithium valproate as described in claim 1 or 2.

6. The use of lithium valproate as described in claim 1 or 2, or the pharmaceutical composition as described in claim 5, in the preparation of a medicament for treating and / or preventing neurological diseases, characterized in that, The neurological disorders mentioned refer to rapid antidepressant and agitation.

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