Use of carbasalate calcium in preparation of drug for treating hydrocephalus

Abstract

The present invention belongs to the technical field of drugs for treating hydrocephalus. Disclosed in the present invention is the use of carbasalate calcium in the preparation of a drug for treating hydrocephalus. In the use of carbasalate calcium in the preparation of a drug for treating hydrocephalus, the drug takes carbasalate calcium as the only or main active ingredient. The present invention finds for the first time the effect of carbasalate calcium in the treatment of hydrocephalus. Carbasalate calcium can significantly reduce the area of the fourth ventricle in a zebrafish model of hydrocephalus, and significantly improve the swimming distance, swimming duration, swimming speed, and rotational movement frequency of the zebrafish model of hydrocephalus. The experimental data provided by the present invention show that carbasalate calcium has great application prospects in the treatment of hydrocephalus, which provides an effective candidate drug for alleviating the suffering of hydrocephalus patients during treatment, and for reducing treatment risks and costs, thereby expanding the application range of carbasalate calcium.

Description

Use of calcium carbapropate in the preparation of a drug for treating hydrocephalus TECHNICAL FIELD

[0001] The present application relates to the technical field of hydrocephalus treatment drugs, in particular to the use of calcium carbapropate in the preparation of a drug for treating hydrocephalus. BACKGROUND

[0002] Hydrocephalus is a condition characterized by the abnormal accumulation of cerebrospinal fluid (CSF) within the skull. It is usually caused by a disturbance in the circulation, absorption, or overproduction of CSF. Hydrocephalus can be classified into two main types: obstructive hydrocephalus and communicating hydrocephalus. Obstructive hydrocephalus occurs when the flow of CSF is blocked, which can be caused by congenital defects (such as Chiari malformation), tumors, infections (such as meningitis), hemorrhages (such as blood clots after subarachnoid hemorrhage), or other inflammatory diseases. The obstruction can occur at various sites within the ventricular system, such as the interventricular foramina or the passage from the ventricles to the subarachnoid space. Communicating hydrocephalus occurs when the flow of CSF is not physically blocked, but rather due to impaired absorption of CSF. This can be caused by inflammation of the meninges, head trauma, decreased absorption capacity of CSF after certain types of brain tumor treatment (such as radiotherapy), or natural aging processes. Additionally, hydrocephalus can be associated with genetic factors, congenital defects, or certain disease states.

[0003] Hydrocephalus can occur at any age, from neonates (infantile hydrocephalus) to the elderly. According to existing statistical data, the incidence of hydrocephalus in children is approximately 1‰, and in the elderly population, the incidence is approximately 2‰. Hydrocephalus can lead to increased intracranial pressure, causing a series of neurological symptoms and signs. The specific symptoms vary depending on the patient's age, cause, and severity of the disease. Common symptoms include headache, vomiting, papilledema (leading to visual problems), difficulty walking, cognitive impairment, and abnormal enlargement of the head circumference in children.

[0004] Traditional methods of treating hydrocephalus mainly include the following: 1) shunt surgery, which is the most common treatment method, a catheter (shunt) is placed in the brain ventricle or outside the brain ventricle through surgery, and the excess cerebrospinal fluid is drained to other parts of the body, usually the abdominal cavity (called ventriculoperitoneal shunt), sometimes to the atrium or chest. A valve is installed on the shunt to regulate the flow rate of cerebrospinal fluid to maintain normal intracranial pressure. 2) Endoscopic surgery, with the development of neuroendoscopic technology, for some types of obstructive hydrocephalus, especially caused by stenosis or obstruction, endoscopic third ventriculostomy (ETV) or endoscopic coagulation of the choroid plexus can be used for treatment. ETV surgery is to create a small hole at the bottom of the third ventricle through an endoscope to allow cerebrospinal fluid to flow directly into the subarachnoid space, thereby restoring normal circulation of cerebrospinal fluid. 3) Drug therapy, although drugs cannot cure hydrocephalus, in some cases, such as mild or temporary hydrocephalus, or as an adjunct to surgery, drugs may be used to reduce cerebrospinal fluid production or help control patient symptoms, and the commonly used drug is a diuretic.

[0005] The treatment methods of shunt surgery and endoscopic surgery both have certain risks and complications, including infection, catheter blockage and excessive drainage. Currently, there is no drug approved by the State Drug Administration for the treatment of human hydrocephalus in clinical practice.

[0006] The main components of carbapropyl calcium are aspirin and calcium salt, which combines the antipyretic, analgesic and anti-inflammatory effects of aspirin and the calcium supplement function, and is mainly used in clinical practice for fever caused by common cold or influenza, and also for relieving mild to moderate pain, such as headache, joint pain, migraine, toothache, muscle pain, neuralgia and menstrual pain. SUMMARY

[0007] The purpose of the present application is to provide the use of carbapropyl calcium in the preparation of a drug for treating hydrocephalus, to solve the problem that current hydrocephalus can only be treated by surgery with high cost, high risk of infection and complications, to provide an effective candidate drug for treating hydrocephalus, and to reduce the pain and risk of treatment for patients and the cost of treatment.

[0008] To achieve the above-mentioned purpose, the present application provides the use of carbapropyl calcium in the preparation of a drug for treating hydrocephalus, wherein the drug contains carbapropyl calcium as the only or main effective component.

[0009] Preferably, the drug further comprises a pharmaceutically acceptable carrier.

[0010] Preferably, the effective amount of carbapropyl calcium is 75-1000 mg / day.

[0011] Preferably, the effective amount of carbapropyl calcium is 150-500 mg / day.

[0012] Preferably, the carbazilicate is used in oral administration in the treatment of hydrocephalus.

[0013] The potential therapeutic target of carbazilicate for hydrocephalus is mmp9.

[0014] The present application uses a zebrafish hydrocephalus model to screen out carbazilicate, which can significantly reduce the fourth ventricle area of hydrocephalus zebrafish, thereby being used for preventing and / or treating hydrocephalus in zebrafish.

[0015] Therefore, the present application provides the use of carbazilicate in the preparation of a drug for treating hydrocephalus, and the specific technical effects are as follows:

[0016] (1) The present application first discovers the effect of carbazilicate in treating hydrocephalus, which can significantly reduce the fourth ventricle area of hydrocephalus model zebrafish, and significantly improve the swimming distance, swimming time, swimming speed and rotation frequency of hydrocephalus model zebrafish.

[0017] (2) The experimental data provided by the present application show that carbazilicate has great application prospects in the treatment of hydrocephalus, which provides an effective candidate drug for reducing the treatment pain of hydrocephalus patients, reducing the treatment risk and cost, and expanding the application range of carbazilicate.

[0018] The technical solutions of the present application will be further described in detail below with the aid of the accompanying drawings and examples. BRIEF DESCRIPTION OF DRAWINGS

[0019] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings needed to be used in the description of the embodiments of the present application will be briefly introduced. Obviously, the drawings in the following description are only some embodiments of the present application, and other drawings can be obtained by those skilled in the art without creative labor.

[0020] Fig. 1 is a schematic diagram of the experimental process of the embodiment one of the present application;

[0021] Fig. 2 is a statistical result of the swimming distance of zebrafish larvae in different treatment groups in the embodiment one of the present application;

[0022] Fig. 3 is a statistical result of the swimming speed of zebrafish larvae in different treatment groups in the embodiment one of the present application;

[0023] Fig. 4 is a statistical result of the swimming time of zebrafish larvae in different treatment groups in the embodiment one of the present application;

[0024] Fig. 5 is a rotation frequency of zebrafish larvae in different treatment groups in the embodiment one of the present application;

[0025] Figure 6 is a statistical result of the swimming distance of the zebrafish larvae in different treatment groups in Example 2 of the present application developed to 4 dpf;

[0026] Figure 7 is a statistical result of the swimming speed of the zebrafish larvae in different treatment groups in Example 2 of the present application developed to 4 dpf;

[0027] Figure 8 is a statistical result of the swimming time of the zebrafish larvae in different treatment groups in Example 2 of the present application developed to 4 dpf;

[0028] Figure 9 is a statistical result of the rotational movement frequency of the zebrafish larvae in different treatment groups in Example 2 of the present application developed to 4 dpf;

[0029] Figure 10 is a statistical result of the swimming distance of the zebrafish larvae in the blank control group, the hydrocephalus group, the hydrocephalus + carbazil calcium group and the carbazil calcium group developed to 4 dpf in Example 2 of the present application;

[0030] Figure 11 is a statistical result of the swimming speed of the zebrafish larvae in the blank control group, the hydrocephalus group, the hydrocephalus + carbazil calcium group and the carbazil calcium group developed to 4 dpf in Example 2 of the present application;

[0031] Figure 12 is a statistical result of the swimming time of the zebrafish larvae in the blank control group, the hydrocephalus group, the hydrocephalus + carbazil calcium group and the carbazil calcium group developed to 4 dpf in Example 2 of the present application;

[0032] Figure 13 is a statistical result of the rotational movement frequency of the zebrafish larvae in the blank control group, the hydrocephalus group, the hydrocephalus + carbazil calcium group and the carbazil calcium group developed to 4 dpf in Example 2 of the present application;

[0033] Figure 14 is a schematic diagram of the experimental process of Example 3 of the present application;

[0034] Figure 15 is a body microscope photograph of the zebrafish larvae in the blank control group (A), the hydrocephalus group (B), the hydrocephalus + carbazil calcium group (C) and the carbazil calcium group (D) in Example 3 of the present application;

[0035] Figure 16 is a statistical result of the fourth ventricle area of the zebrafish larvae in the blank control group, the hydrocephalus group, the hydrocephalus + carbazil calcium group and the carbazil calcium group developed to 72 hpf in Example 3 of the present application;

[0036] Figure 17 is a statistical result of the number of apoptotic cells in the fourth ventricle of the zebrafish larvae in the blank control group, the hydrocephalus group, the hydrocephalus + carbazil calcium group and the carbazil calcium group developed to 72 hpf in Example 4 of the present application;

[0037] Figure 18 is a Venn diagram of the drug-related target points and the hydrocephalus-related target points in Example 5 of the present application;

[0038] Figure 19 is a statistical result of the fourth ventricle area of the zebrafish larvae in the blank control group, the hydrocephalus group, the hydrocephalus + MMP-9-IN-9 group and the MMP-9-IN-9 group in Example Five of the present application developed to 72hpf.

[0039] Figure 20 is a statistical result of the fourth ventricle area of the zebrafish larvae in the blank control group, the hydrocephalus group, the hydrocephalus + MMP-9-IN-1 group and the MMP-9-IN-1 group in Example Five of the present application developed to 72hpf.

[0040] Figure 21 is a statistical result of the ratio of the ventricle volume to the whole brain volume of the rats in the sham operation group, the hydrocephalus group, the hydrocephalus + carbapropidine calcium nasal drop group and the hydrocephalus + carbapropidine calcium nasal spray group in Example Six of the present application.

[0041] Figure 22 is a magnetic resonance imaging result of the rats in the sham operation group, the hydrocephalus group, the hydrocephalus + carbapropidine calcium nasal drop group and the hydrocephalus + carbapropidine calcium nasal spray group in Example Six of the present application. DETAILED DESCRIPTION

[0042] The technical solutions of the present application are further described below by means of the accompanying drawings and examples.

[0043] In order to make the purpose, technical solutions and advantages of the present application more clear, thorough and complete, the technical solutions of the present application are described clearly and completely below by means of the accompanying drawings and examples. The following detailed description is a description of the examples, which aims to provide further detailed description of the present application. Unless otherwise specified, all technical terms used in the present application have the same meaning as generally understood by those skilled in the art to which the present application belongs.

[0044] The instrument equipment and reagent materials used in the examples are obtained through commercial channels, wherein the zebrafish larvae used are zebrafish embryos born 6h; GRI977143 is purchased from Med Chem Express Company, carbapropidine calcium is purchased from Med Chem Express Company, tricaine is purchased from SIGMA Company, PTU is purchased from SIGMA Company, AO stock solution is purchased from SIGMA Company, MS-222 is purchased from Med Chem Express Company, a body microscope is purchased from Nikon Company, a zebrafish behavior recorder is purchased from Noldus Company, and an upright fluorescence microscope is purchased from OLYMPUS Company.

[0045] The experimental steps not described in detail in the examples are conventional methods in the art.

[0046] Example One

[0047] The effect of carbapropidine calcium on hydrocephalus of zebrafish larvae was investigated, and the flowchart is shown in Figure 1, and the specific steps are as follows:

[0048] (1) Preparation of zebrafish larvae

[0049] Wild-type zebrafish were obtained from the China Zebrafish Resource Center and routinely maintained at 28±0.5℃ under 14 h light / 10 h dark conditions. Zebrafish embryos were obtained by natural mating of wild-type zebrafish, and the obtained zebrafish embryos were washed and stored in E3 medium containing 2 mg / L methylene blue, during which the pH of the medium was maintained at 7.1 and the medium was replaced daily.

[0050] The E3 medium consisted of 5 mM NaCl, 0.17 mM KCl, 0.33 mM CaCl2, and 0.33 mM MgSO4.

[0051] (2) Experimental grouping and treatment

[0052] a. Blank control group: 6 hpf zebrafish embryos were randomly transferred to a 24-well cell culture plate, 15 zebrafish embryos per well, 1 mL of E3 medium was added, so that all 6hpf zebrafish embryos were in E3 medium, and the medium was replaced every 24 h.

[0053] b. Hydrocephalus group (Hydro): 6hpf zebrafish embryos were randomly transferred to a 24-well cell culture plate, 15 6hpf zebrafish embryos per well, then 5 μg / mL GRI977143 solution was added, so that all 6hpf zebrafish embryos were exposed to the solution to induce zebrafish hydrocephalus, and the GRI977143 solution was replaced every 24 h.

[0054] c. Hydrocephalus + carbaplatin calcium group: 6hpf zebrafish embryos were randomly transferred to a 24-well cell culture plate, 15 6hpf zebrafish embryos per well, then 5 μg / mL GRI977143 solution was added, so that all 6hpf zebrafish embryos were exposed to the solution, and when the embryos developed to 24 h (24hpf), carbaplatin calcium solution was added to make the carbaplatin calcium concentration 5 μM, 10 μM, and 20 μM, respectively, and the GRI977143 solution concentration was maintained at 5 μg / mL, and the solution was replaced every 24 h.

[0055] d. Carbaplatin calcium group: 6hpf zebrafish embryos were randomly transferred to a 24-well cell culture plate, 15 6hpf zebrafish embryos per well, and when the embryos developed to 24 h (24hpf), carbaplatin calcium solution was added to make the carbaplatin calcium concentration 5 μM, 10 μM, and 20 μM, respectively, and the solution was replaced every 24 h.

[0056] (3) Zebrafish behavioral detection

[0057] The zebrafish larvae of each group were detected by zebrafish behavior when the embryos developed to 72 h (72hpf), and the zebrafish embryos developed to 4 days (4dpf) were placed in a 48-well plate, one in each well, and 1000 μL of E3 culture solution was added. After adapting for 20 min using DanioVision (Noldus) recorder, 1 h was recorded in 10 min / 10 min light / dark cycle. The statistical results of the swimming distance, swimming time, swimming speed and rotation frequency of 4dpf zebrafish larvae are shown in Figures 2-5, respectively. The swimming distance, swimming time, swimming speed and rotation frequency of the hydrocephalus group of zebrafish were significantly decreased compared with the blank control group; after treatment with carbaplatin calcium (20 μM), the swimming distance, swimming time, swimming speed and rotation frequency were significantly increased; the swimming distance, swimming time, swimming speed and rotation frequency of the zebrafish in the group without inducing hydrocephalus and adding carbaplatin calcium had no significant difference compared with the blank control group.

[0058] Example Two

[0059] The optimal treatment concentration of carbaplatin calcium solution was investigated, and the specific steps were as follows:

[0060] (1) The preparation of zebrafish larvae was the same as in Example One.

[0061] (2) Experimental grouping and treatment

[0062] a. Blank control group: 6 h (6hpf) old zebrafish embryos were randomly transferred to a 24-well cell culture plate, 15 zebrafish embryos in each well, and 1 mL of E3 culture solution was added to make all 6hpf zebrafish embryos in the E3 culture solution, and the culture solution was replaced every 24 h.

[0063] b. Hydrocephalus group (Hydro): 6hpf zebrafish embryos were randomly transferred to a 24-well cell culture plate, 15 6hpf zebrafish embryos in each well, and then 5 μg / mL of GRI977143 solution was added to expose all 6hpf zebrafish embryos to the solution to induce zebrafish hydrocephalus, and the GRI977143 solution was replaced every 24 h.

[0064] c. Hydrocephalus + Carbaplatin group: 6hpf zebrafish embryos were randomly transferred into 24-well cell culture plates, 15 6hpf zebrafish embryos in each well, then 5 μg / mL GRI977143 solution was added to expose all 6hpf zebrafish embryos to the solution, and when the embryos developed to 24 h (24hpf), carbaplatin solution was added to make the carbaplatin concentration 10 μM, 15 μM, 20 μM, 25 μM respectively, and the concentration of GRI977143 solution was kept at 5 μg / mL, and the solution was replaced every 24 h.

[0065] d. Carbaplatin group: 6hpf zebrafish embryos were randomly transferred into 24-well cell culture plates, 15 6hpf zebrafish embryos in each well, and when the embryos developed to 24 h (24hpf), carbaplatin solution was added to make the carbaplatin concentration 10 μM, 15 μM, 20 μM, 25 μM respectively, and the solution was replaced every 24 h.

[0066] (3) Zebrafish behavior test same as Example One

[0067] The results of the swimming distance, swimming time, swimming speed and rotation frequency of 4dpf zebrafish larvae are shown in Figures 6-9 respectively. The swimming distance, swimming time, swimming speed and rotation frequency of the hydrocephalus group zebrafish were significantly decreased compared with the blank control group. Compared with the 10 μM, 15 μM, 25 μM carbaplatin concentration treatment group of zebrafish larvae, the swimming distance, swimming time, swimming speed and rotation frequency of zebrafish larvae treated with 20 μM carbaplatin increased more obviously. There was no significant difference in the swimming distance, swimming time, swimming speed and rotation frequency of the zebrafish in the group without inducing hydrocephalus but adding 20 μM carbaplatin compared with the blank control group.

[0068] Therefore, the optimal dosage concentration of carbaplatin is 20 μM. The results of the swimming distance, swimming time, swimming speed and rotation frequency of zebrafish larvae at the optimal dosage concentration of carbaplatin are shown in Figures 10-13 respectively.

[0069] Example Three

[0070] The ventricle of zebrafish was imaged by a body microscope, and the specific steps were as follows:

[0071] (1) Preparation of zebrafish larvae same as Example One.

[0072] (2) The experimental grouping and treatment are the same as in Example Two, and E3 culture solution is replaced with PTU solution. PTU (phenylthiourea) is a Tyr (tyrosinase) inhibitor that can inhibit the growth of melanin in zebrafish embryos, increase the optical transparency of zebrafish embryos, and facilitate the taking of better images under a microscope for research. The concentration of carbaplatin calcium in the hydrocephalus + carbaplatin calcium group and the carbaplatin calcium group is 20 μM.

[0073] (3) Zebrafish ventricle imaging

[0074] When the zebrafish embryos develop to 72 h (72hpf), the embryos in each group are embedded with low-melting-point agarose and then used. After the stereomicroscope is adjusted, the images of the zebrafish ventricles are obtained, and the flowchart is shown in FIG. 14. Some stereomicroscope photos are shown in FIG. 15. The size of the ventricle is measured using Image J software, and the statistical results of 13 repetitions are shown in FIG. 16. The fourth ventricle area of the zebrafish in the hydrocephalus group is significantly increased compared with the blank control group. After treatment with carbaplatin calcium, the fourth ventricle area is significantly restored. The zebrafish in the group without induced hydrocephalus but with added carbaplatin calcium has no significant difference in the fourth ventricle area compared with the blank control group.

[0075] Example Four

[0076] The number of apoptotic cells in the fourth ventricle of zebrafish is counted using an upright fluorescence microscope, and the specific steps are as follows:

[0077] (1) The preparation of zebrafish larvae is the same as in Example One.

[0078] (2) The experimental grouping and treatment are the same as in Example Three.

[0079] (3) Zebrafish fourth ventricle apoptosis cell number statistics

[0080] Acridine orange (AO) is a staining agent for identifying apoptosis. AO stock solution (1 mg / mL, prepared with pure water) was diluted with E3 medium at a ratio of 1:200 to make its final concentration 5 μg / mL. Twelve zebrafish larvae (72hpf) were randomly selected from different treatment groups, rinsed with E3 medium to remove residual exposure solution, and placed in centrifuge tubes, followed by the addition of 1 mL of diluted AO dye. Incubate at 28±0.5 ℃ for 20 minutes and keep in the dark to prevent fluorescence quenching. After incubation, wash with E3 solution for 5 minutes to remove AO dye that has not entered the cells, and then anesthetize the larvae with 0.01% MS-222 for 3 minutes. Take pictures of apoptotic cells in zebrafish in vivo (filter: GFP, wavelength: 470±20 nm) using an upright fluorescence microscope. Use Imaris to count the number of apoptotic cells in the fourth ventricle of each larva. The statistical results of 12 replicates are shown in Figure 17. The number of apoptotic cells in the fourth ventricle of zebrafish in the hydrocephalus group increased significantly compared with the blank control group; after treatment with carbaplatin calcium, the number of apoptotic cells decreased; there was no significant difference in the number of apoptotic cells in the fourth ventricle of zebrafish in the group with no induced hydrocephalus but added carbaplatin calcium compared with the blank control group.

[0081] Example Five

[0082] Determination of potential therapeutic targets of carbaplatin calcium for hydrocephalus, the specific steps are as follows:

[0083] In GeneCards (https: / / www.genecards.org / ), OMIM (https: / / www.omim.org / ) and DisGeNET (https: / / disgenet.com / ), the keyword "Hydrocephalus" was used to search for hydrocephalus-related targets, the search results of the three databases were integrated and duplicate gene targets were removed to obtain hydrocephalus-related targets, and a hydrocephalus disease target library was constructed.

[0084] The blank control group and hydrocephalus group zebrafish larvae samples were sent to Baimaik company for transcriptome sequencing analysis to obtain the differentially expressed genes of the blank control group and the hydrocephalus group, and a transcriptome target library was constructed.

[0085] Drug-related targets were retrieved using the keywords “GRI977143” and “Carbasalatum calcicum” in SwissTargetPrediction (http: / / swisstargetprediction.ch / index.php), PharmMapping (https: / / www.lilab-ecust.cn / pharmmapper / ), and the Similarity ensemble approach (http: / / sea.bkslab.org / ). The search results from the three databases were integrated and duplicate gene targets were removed to obtain drug-related targets, and a GRI977143 target library and a carbaspirin calcium target library were constructed.

[0086] By importing drug-related targets and hydrocephalus-related targets into the Venny 2.1.0 web platform (https: / / bioinfogp.cnb.csic.es / tools / venny / index.html) to generate a Venn diagram, as shown in Figure 18, the intersection target was found to be mmp9.

[0087] Subsequently, the mmp9 inhibitor was applied to juvenile zebrafish to verify whether mmp9 is a target for carbaspirin calcium therapy. The specific steps are as follows:

[0088] (1) The preparation of zebrafish fry is the same as in Example 1.

[0089] (2) The experimental groups and treatments were the same as in Example 2, except that carbaspirin calcium was replaced with MMP9 inhibitors (MMP-9-IN-9, MMP-9-IN-1).

[0090] (3) Imaging of the zebrafish ventricles with a stereomicroscope is the same as in Example 3.

[0091] Compared with the blank control group, the area of ​​the fourth ventricle in zebrafish in the hydrocephalus group was significantly increased; after treatment with MMP inhibitors, the area of ​​the fourth ventricle was significantly restored, as shown in Figures 19-20.

[0092] Therefore, the potential therapeutic target of carbaspirin calcium is MMP9.

[0093] Therefore, the application firstly discovers the effect of carbazilamide calcium in treating hydrocephalus, can significantly reduce the fourth ventricle area of the hydrocephalus model zebra fish, significantly improve the swimming distance, swimming time, swimming speed and rotating motion frequency of the hydrocephalus model zebra fish, and significantly reduce the number of apoptotic cells in the fourth ventricle of the hydrocephalus model zebra fish; the potential therapeutic target is mmp9. The experimental data provided by the application shows that carbazilamide calcium has great application prospect in the treatment of hydrocephalus, provides an effective candidate drug for reducing the treatment pain of the hydrocephalus patient, reducing the treatment risk and cost, and expands the application range of carbazilamide calcium.

[0094] Example six

[0095] The operation is carried out under sterile conditions on rats anesthetized with isoflurane, and the body temperature of the rats is maintained at 37℃ using a heating pad. The rats are placed in a stereotaxic frame (Rveod) and the skull is exposed through a midline incision, and a hole is drilled on the left lateral ventricle (0.6 mm behind the bregma and 1.6 mm lateral to the bregma) using a skull drill. Then, within 30 minutes, a 33G needle is inserted into the left lateral ventricle (4.5 mm ventrally) through the drill hole by a sample injection needle (Hamilton Instruments) to inject 100 μl of LPA2 receptor activator (hydrocephalus group) or normal saline (sham operation group), which is expected to diffuse throughout the ventricular system. The needle is kept in place for 5 minutes before being withdrawn to prevent backflow. The skin incision is sutured with suture thread, and the rats are returned to the feeding facility after recovery. The sham operation group and the hydrocephalus group are not subjected to special treatment. The carbazilamide calcium nasal treatment group (hydrocephalus + carbazilamide calcium nasal treatment) is treated 3 days after the operation. The treatment process is as follows: after the rats are anesthetized, the rats are kept in a supine position with the neck slightly raised to 45°. A total dose of 30 mg / kg / d is administered alternately to both nostrils, with a volume of 20 μL per nostril each time, and an interval is left between the two administrations to ensure absorption of the liquid medicine. This operation is performed twice a day, with a single operation time of about 20 minutes, and lasts for 14 days. The carbazilamide calcium nasal spray treatment group (hydrocephalus + carbazilamide calcium nasal spray) is anesthetized, and the rats are kept in a supine position with the neck slightly raised to 45°. The liquid medicine is loaded with an Aptar sprayer (Aptar). A total dose of 30 mg / kg / d is administered alternately to both nostrils, with a volume of 20 μL per nostril each time, and an interval is left between the two administrations to ensure absorption of the liquid medicine. This operation is performed twice a day, with a single operation time of about 20 minutes, and lasts for 14 days.

[0096] The composition of the nasal spray liquid is as follows.

[0097]

[0098] After two weeks of drug administration, MRI was performed on rats in all groups. The ventricle expansion phenomenon was evaluated by calculating the ratio of the ventricle volume to the whole brain volume. The results of MRI image analysis are shown in Figures 21-22. The rats in the hydrocephalus group showed significant ventricle expansion compared to the blank control group, showing typical hydrocephalus characteristics, and the difference was statistically significant. The degree of ventricle expansion in the carbaplatin calcium nasal drop treatment group was significantly lower than that in the hydrocephalus group, and the difference was statistically significant, indicating that carbaplatin calcium nasal drops had a therapeutic effect on communicating hydrocephalus in rats. The degree of ventricle expansion in the carbaplatin calcium nasal spray treatment group was significantly lower than that in the hydrocephalus group, and the difference was statistically significant, indicating that carbaplatin calcium nasal spray had a therapeutic effect on communicating hydrocephalus in rats.

[0099] Finally, it should be noted that the above examples are only used to illustrate the technical solutions of the present application and not to limit it. Although the present application has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solutions of the present application can still be modified or replaced by equivalents, and these modifications or replacements cannot make the modified technical solutions deviate from the spirit and scope of the technical solutions of the present application.

Claims

1. Use of carbamazepine calcium for the preparation of a medicament for the treatment of hydrocephalus, characterized in that: The medicine takes carbazochrome calcium as the only or main effective component.

2. Use of the carbamazepine calcium according to claim 1 for the preparation of a medicament for the treatment of hydrocephalus, characterized in that: The medicine also includes a pharmaceutically acceptable carrier.

3. Use of the carbamazepine calcium according to claim 1 for the preparation of a medicament for the treatment of hydrocephalus, characterized in that: The effective amount of the carbazochrome calcium is 75-1000 mg / day.

4. Use of the carbamazepine calcium according to claim 1 for the preparation of a medicament for the treatment of hydrocephalus, characterized in that: The effective amount of the carbazochrome calcium is 150-500 mg / day.

5. Use of the carbamazepine calcium according to claim 1 for the preparation of a medicament for the treatment of hydrocephalus, characterized in that: The carbazochrome calcium is used in oral way in treating hydrocephalus.

6. Use of the carbamazepine calcium according to claim 1 for the preparation of a medicament for the treatment of hydrocephalus, characterized in that: The carbazochrome calcium is used in nasal drop way in treating hydrocephalus.

7. Use of the carbamazepine calcium according to claim 1 for the preparation of a medicament for the treatment of hydrocephalus, characterized in that: The carbazochrome calcium is used in nasal spray way in treating hydrocephalus.

8. The potential therapeutic target of carbazochrome calcium for hydrocephalus is mmp9.

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