Use of bicyclohexanone oxalyl dihydrazone in a drug for stroke and a drug for stroke

Abstract

This invention provides the use of dicyclohexanone oxaloyl dihydrazone in stroke medications and a stroke medication itself. The use includes the application of dicyclohexanone oxaloyl dihydrazone in the preparation of medications for treating stroke, particularly ischemic stroke. This invention effectively promotes myelin regeneration and neurological function repair in the very early post-stroke period (e.g., within 24 hours) by targeting and regulating the "ceruloplasmin-iron metabolism-myelin regeneration" axis. The specific mechanisms include upregulating ceruloplasmin expression in the brain, enhancing its ferrooxidase activity, promoting iron transport and reuse in the brain, thereby promoting the survival, differentiation, and myelination of oligodendrocyte precursor cells. Animal experiments show that stroke model mice exhibit significant improvement in neurological function as early as day 1 after administration, with a faster onset of action than existing technologies. This invention has the advantages of rapid onset of action, clear target, and comprehensive repair effects, and has promising clinical application prospects.

Description

Technical Field

[0001] This invention relates to pharmaceutical technology, and more particularly to the use of dicyclohexanone oxalyl dihydrazone in stroke medications and stroke medications. Background Technology

[0002] Stroke is one of the leading causes of disability and death worldwide, with ischemic stroke accounting for the highest proportion. Currently, clinical treatment mainly focuses on acute-phase vascular recanalization (such as thrombolysis and mechanical thrombectomy) and neuroprotection, aiming to restore blood flow as quickly as possible and salvage the ischemic penumbra. However, there are currently no specific drugs for the repair of neurological function after stroke, especially for promoting white matter damage repair, myelin regeneration, and neural circuit reconstruction.

[0003] Differentiation and myelination of oligodendrocyte precursor cells (OPCs) are crucial steps in neurological repair following central nervous system injury. After stroke, the differentiation of OPCs into mature oligodendrocytes is impaired, leading to impaired axonal myelination, a significant cause of persistent neurological deficits. Studies have shown that abnormal iron metabolism is closely related to post-stroke oxidative stress, inflammatory responses, and OPC dysfunction. Disruption of iron homeostasis in the brain, especially the excretion of toxic ferrous ions (Fe2+), is a major contributing factor. 2+ Excessive accumulation of OPCs can generate a large amount of reactive oxygen species through the Fenton reaction, exacerbating white matter damage and inhibiting the survival and differentiation of OPCs.

[0004] Ceruloplasmin (CP) is an important iron-oxidizing enzyme secreted by astrocytes in the brain, responsible for converting toxic iron... 2+ Oxidized into ferric ions (Fe3+) that can be bound by transport proteins. 3+ Ceruloplasmin plays a central role in maintaining brain iron homeostasis. However, the mechanism of action of ceruloplasmin in white matter repair after ischemic stroke remains unclear, and there are currently no drugs that target the ceruloplasmin-iron metabolic axis to promote myelin regeneration and are available for clinical use.

[0005] In existing technologies, some studies have attempted to promote post-stroke neurological function recovery through other pathways. For example, existing technology (Li-Hsin Chang et al., Blockade of soluble epoxide hydrolase attenuates post-ischemic neuronal hyperexcitation and confers resilience against stroke with TrkB activation) discloses a method for treating ischemic stroke by inhibiting soluble epoxide hydrolase (sEH). This technology uses the sEH inhibitor AUDA, which enhances neurogenesis and synaptic plasticity by reducing post-ischemic neuronal hyperexcitation and activating the TrkB signaling pathway, showing effects of reducing infarct volume and improving motor function in animal models.

[0006] However, the existing technology has the following drawbacks: (1) Its mechanism of action mainly focuses on promoting neuronal survival and synaptic remodeling, and its effect on the equally important white matter damage repair and myelin regeneration process after stroke is limited or not mentioned; (2) It does not involve the regulatory role of the ceruloplasmin-iron metabolism axis in post-stroke repair, so it cannot effectively intervene in the core pathological link of OPCs differentiation disorder and white matter repair obstruction caused by iron metabolism disorder; (3) Its behavioral function recovery effect usually appears several days after stroke (such as 7 days after surgery), and its effect is slow, making it difficult to meet the needs of rapid initiation of neuronal repair after the acute phase in clinical practice.

[0007] Therefore, there is an urgent need in this field to develop a drug or treatment strategy that can effectively promote myelin regeneration and neurological function repair in the very early post-stroke period by regulating ceruloplasmin expression and brain iron metabolism. Summary of the Invention

[0008] The purpose of this invention is to address the current limitations of drugs in intervening in the core pathological aspects of white matter damage and their slow onset of action. This invention proposes the use of bicyclohexanone oxaloyl dihydrazone in the preparation of stroke drugs. By using a novel treatment strategy that targets and regulates the ceruloplasmin-iron metabolism axis with bicyclohexanone oxaloyl dihydrazone, myelin regeneration and neurological function repair can be effectively promoted in the very early stages after stroke.

[0009] It should be noted that, in this invention, unless otherwise specified, the specific meaning of "comprising" in relation to composition and description includes both open-ended meanings such as "comprising," "including," etc., and closed-ended meanings such as "composed of," "consisting of," etc., and similar meanings.

[0010] To achieve the above objectives, the technical solution adopted by the present invention is: the use of cuprizone (CPZ) in the preparation of stroke drugs.

[0011] Furthermore, the stroke is an ischemic stroke.

[0012] Furthermore, the drug is administered within 24 hours after the stroke, preferably within 12 hours, and more preferably within 1 hour.

[0013] Furthermore, the effective dosage of the dicyclohexanone oxalyl dihydrazone is 10-20 mg / kg daily, preferably 10-15 mg / kg daily. The duration of treatment is 3-14 days.

[0014] Furthermore, the bicyclohexanone oxaloyl dihydrazone achieves its use through at least one of the following mechanisms:

[0015] a) Upregulates the expression of ceruloplasmin in the brain;

[0016] b) Enhances the activity of ferrooxidase in ceruloplasmin in the brain;

[0017] c) Promotes the transport or reuse of iron in the brain;

[0018] d) Promotes the survival, differentiation, or myelination of oligodendrocyte precursor cells;

[0019] e) Promotes the repair of neurological function after stroke.

[0020] Another object of the present invention discloses a medicament (for treating) stroke, comprising dicyclohexanone oxalyl dihydrazone, or a pharmaceutically acceptable salt, ester, or prodrug thereof.

[0021] Further, the pharmaceutically acceptable salt of the dicyclohexanone oxaloyl dihydrazone is selected from acid addition salts or base addition salts; the acid addition salts include salts formed with inorganic or organic acids, wherein the inorganic acids include, but are not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, and phosphoric acid; the organic acids include, but are not limited to, formic acid, acetic acid, trifluoroacetic acid, propionic acid, oxalic acid, maleic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, methanesulfonic acid, benzenesulfonic acid, and p-toluenesulfonic acid; the base addition salts include salts formed with inorganic or organic bases, wherein the inorganic bases include, but are not limited to, hydroxides or carbonates of sodium, potassium, lithium, calcium, magnesium, and aluminum; the organic bases include, but are not limited to, ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, ethanolamine, lysine, arginine, histidine, and choline.

[0022] Furthermore, the ester is a carboxylic acid ester of dicyclohexanone oxaloyl dihydrazone, including but not limited to methyl ester, ethyl ester, propyl ester, isopropyl ester, butyl ester, and tert-butyl ester; or an amino acid ester, including but not limited to glycine ester, alanine ester, valine ester, and leucine ester.

[0023] Furthermore, the prodrug includes derivatives that release dicyclohexanone oxaloyl dihydrazone in vivo via enzymatic or chemical conversion, including but not limited to ester prodrugs, amide prodrugs, phosphate prodrugs, or azo-reduction activated prodrugs.

[0024] Furthermore, the stroke is an ischemic stroke.

[0025] Furthermore, the route of administration for the stroke drug is intravenous injection.

[0026] Furthermore, the stroke medication is administered within 24 hours of the stroke.

[0027] Furthermore, the stroke medication can be used in combination with existing thrombolytic drugs, neuroprotective agents, or rehabilitation training to form a synergistic treatment plan.

[0028] Furthermore, the stroke medication also includes pharmaceutically acceptable carriers, excipients, or diluents. Including but not limited to: fillers / diluents: lactose, microcrystalline cellulose, mannitol, starch, pregelatinized starch, dicalcium phosphate; binders: hydroxypropyl methylcellulose, povidone, ethyl cellulose, gelatin, gum arabic; disintegrants: croscarmellose sodium, croscarmellose, carboxymethyl starch sodium, low-substituted hydroxypropyl cellulose; lubricants / flow aids: magnesium stearate, talc, polyethylene glycol, colloidal silica, stearic acid; solvents / solvents: water for injection, ethanol, propylene glycol, polyethylene glycol 400, dimethyl sulfoxide; surfactants: Tween 80, sodium dodecyl sulfate, poloxamer; preservatives: benzyl alcohol, parabens, phenol; isotonic adjusters: sodium chloride, glucose, glycerol; pH adjusters: hydrochloric acid, sodium hydroxide, citric acid, phosphate buffer; antioxidants: sodium sulfite, sodium metabisulfite, vitamin E, butylated hydroxyanisole.

[0029] The use of dicyclohexanone oxalyl dihydrazone in stroke medications and the stroke medications of this invention have the following advantages compared with the prior art:

[0030] 1) This invention clarifies that regulating the ceruloplasmin-iron metabolism axis through bicyclohexanone oxaloyl dihydrazone (CPZ) is a novel strategy for treating stroke. This strategy directly targets the core pathological link of post-stroke iron metabolism disorder leading to impaired differentiation of oligodendrocyte precursor cells and hindered white matter repair, thus solving the problem that existing technologies have limited effects on myelin regeneration or do not involve the regulation of iron metabolism.

[0031] 2) The technical solution of this invention aims to initiate the repair process in the very early stage after stroke (e.g., within 24 hours). According to specific implementation methods, animal experiments have shown that the neurological function score of the CPZ treatment group was significantly better than that of the model control group on the first day after stroke, indicating that it can quickly produce behavioral improvement and has a faster onset of action than existing technologies (such as AUDA mentioned in the comparative document, which usually shows functional recovery effects several days after surgery).

[0032] 3) This invention promotes iron transport and reuse in the brain by upregulating ceruloplasmin expression and enhancing its ferrooxidase activity, thereby creating a favorable microenvironment for the survival, differentiation, and myelination of oligodendrocyte precursor cells. This not only promotes white matter (myelin) regeneration but also globally reduces secondary damage by improving iron homeostasis in the brain, achieving more comprehensive neural repair.

[0033] 4) This invention provides the use of dicyclohexanone oxalyl dihydrazone or its pharmaceutically acceptable salts, esters, or prodrugs in the preparation of drugs for stroke (especially ischemic stroke), and provides specific timing of administration (e.g. within 24 hours after stroke), route of administration (e.g., intravenous injection) and dosage range (e.g., 10 mg / kg daily), forming a complete and feasible treatment regimen.

[0034] The most significant advantage of this invention lies in its innovative targeting of the novel pathway of "ceruloplasmin-iron metabolism-myelin regeneration," which enables rapid and effective promotion of myelin regeneration and neurological function repair in the very early stages after stroke, filling the gaps in existing treatments for white matter damage repair after stroke. Attached Figure Description

[0035] Figure 1 The images show immunofluorescence staining of ceruloplasmin, including Con 1dm stroke 1-day control group, CPZ 1dm stroke 1-day treatment group, Con 7dm stroke 7-day control group, CPZ 7dm stroke 7-day treatment group, CP ceruloplasmin, GFAP glial fibrillary acidic protein, Dapi nuclear markers, and Merge fitting plot.

[0036] Figure 2 The images show protein immunoblotting patterns. The mouse distal middle cerebral artery occlusion model group was prepared using dMCAO microscopic electrocoagulation, the dMCAO+CPZ stroke model group was treated, the CS control side was observed, the IS lesion side was observed, GFAP (glial fibrillary acidic protein), FPN1 (membrane iron transporter 1), CP (ceruloplasmin), MBP (myelin basic protein), TfR1 (transferrin receptor 1), olig2 (oligodendrocyte transcription factor 2), FtL (ferritin light chain), and β-actin (actin internal control).

[0037] Figure 3 for Figure 2 Protein expression statistics;

[0038] Figure 4 Mass spectrometry images of iron in the brain of stroke mice after administration: Con 1dm stroke 1 day control group, CPZ 1dm stroke 1 day administration group, Con 7dm stroke 7 day control group, and CPZ 7dm stroke 7 day administration group.

[0039] Figure 5 For ICP-MS determination of the total content of metal elements Cu, Zn, and Fe on the damaged side and the control side, *p < 0.05;

[0040] Figure 6 Immunofluorescence staining of oligodendrocyte precursor cells in control mice one day after stroke; oligodendrocyte transcription factor 2 (O2), Dapi nuclear markers, Merge fitting plot; CS control side, IS lesion side;

[0041] Figure 7 Immunofluorescence staining of oligodendrocyte precursor cells in mice treated one day after stroke; oligodendrocyte transcription factor 2, Dapi nuclear markers, Merge fitting plot; CS control side, IS lesion side;

[0042] Figure 8 Immunofluorescence staining of oligodendrocyte precursor cells in control mice seven days after stroke; oligodendrocyte transcription factor 2 (O2), Dapi nuclear markers, Merge fitting plot; CS control side, IS lesion side;

[0043] Figure 9 Immunofluorescence staining of oligodendrocyte precursor cells in mice treated for seven days after stroke; oligodendrocyte transcription factor 2, Dapi nuclear markers, Merge fitting plot; CS control side, IS lesion side;

[0044] Figure 10 for Figure 6-9 The fluorescence intensity statistics are shown in the following figures: Con 1dm stroke 1-day control group, CPZ 1dm stroke 1-day administration group, Con 7dm stroke 7-day control group, CPZ 7dm stroke 7-day administration group, Cs control side, Is lesion side, **p < 0.01, ***p < 0.001;

[0045] Figure 11Immunofluorescence staining images of mature oligodendrocytes and ferritin light chains are shown. The groups include the Con 1dm stroke-day 1 control group, the CPZ 1dm stroke-day 1 treatment group, the Con 7dm stroke-day 7 control group, the CPZ 7dm stroke-day 7 treatment group, and specific markers for the maturation stage of CC1 oligodendrocytes, FTL ferritin light chains. *p < 0.05, **p < 0.01, ***p < 0.001.

[0046] Figure 12 for Figure 11 The relative number of CC1 positive cells fluorescence intensity statistics, where Con 1dm stroke 1 day control group, CPZ 1dm stroke 1 day administration group, Con 7dm stroke 7 day control group, CPZ 7dm stroke 7 day administration group, Cs control side, Is lesion side, **p < 0.01, ***p < 0.001;

[0047] Figure 13 for Figure 11 The relative number of FTL-positive cells and fluorescence intensity statistics are shown in the following figures: Con 1dm stroke 1-day control group, CPZ 1dm stroke 1-day treatment group, Con 7dm stroke 7-day control group, CPZ 7dm stroke 7-day treatment group, Cs control side, Is injury side, **p < 0.01, ***p < 0.001;

[0048] Figure 14 This is a schematic diagram of gait analysis behavioral scores. Con control group, CPZ-treated group, *p < 0.05, **p < 0.01, ***p < 0.00;

[0049] Figure 15 This is a schematic diagram of TTC staining results, showing the Con control group and the CPZ-treated group.

[0050] Figure 16 for Figure 10 TTC staining lesion volume statistics, Con control group, CPZ treatment group *p < 0.05, **p < 0.01, ***p < 0.001. Detailed Implementation

[0051] The present invention will be further described below with reference to embodiments. The description of the technical features described below is based on representative embodiments and specific examples of the present invention, but the present invention is not limited to these embodiments and specific examples. It should be noted that:

[0052] Unless otherwise stated, all units used in this specification are international standard units, and all numerical values ​​and ranges appearing in this invention should be understood to include systematic errors that are unavoidable in industrial production.

[0053] In this specification, the range of values ​​referred to as "value A to value B" refers to the range including the endpoint values ​​A and B.

[0054] In this specification, the numerical range indicated by "above" or "below" refers to the numerical range that includes the stated number.

[0055] In this specification, the word "may" has two meanings: to perform a certain process and not to perform a certain process.

[0056] In this specification, the terms "optional" or "optional" are used to indicate the use or omission of certain substances, components, procedures, application conditions, etc.

[0057] In this instruction manual, when "room temperature" or "room temperature" is used, the temperature can be 15-25℃.

[0058] Unless otherwise specified, all reagents or instruments used in this instruction manual are commercially available products.

[0059] Example 1: Evaluation of the efficacy of dicyclohexanone oxaloyl dihydrazone in the treatment of stroke (mouse model)

[0060] 1. Materials and Methods

[0061] 1.1 Laboratory Animals and Grouping

[0062] Adult C57BL / 6J mice were selected. The mice were randomly divided into three groups:

[0063] Sham surgery group: only the blood vessels are exposed, without any blockage.

[0064] CPZ treatment group: A middle cerebral artery occlusion model was established, and dicyclohexanone oxaloyl dihydrazone (solvent: physiological saline) was injected into the tail vein after surgery at a dose of 10 mg / kg.

[0065] Model control group: A middle cerebral artery occlusion model was established, and the same volume of solvent as the CPZ treatment group was injected into the tail vein after surgery.

[0066] 1.2 Model Establishment

[0067] A distal occlusion model of the middle cerebral artery was established using electrocoagulation. Specifically, after anesthetizing mice, the left middle cerebral artery was exposed via a subtemporal craniotomy, and a miniature electrocoagulator was used to permanently coagulate and block it, thereby inducing focal cerebral ischemia.

[0068] 1.3 Dosing regimen

[0069] Both the CPZ treatment group and the model control group received their first dose via tail vein immediately after successful model establishment (0 hours). Subsequent administration was once daily for 7 consecutive days. The sham surgery group received no drug treatment.

[0070] 1.4 Detection Indicators and Methods

[0071] 1.4.1 Neurological Function Scores

[0072] Motor function of mice was assessed using a gait analysis system on postoperative days 1, 7, and 14. Gait parameters, such as stride length of the left forelimb, left hindlimb, right forelimb, and right hindlimb, were recorded and analyzed, and statistical analysis was performed.

[0073] 1.4.2 Measurement of cerebral infarction volume

[0074] On postoperative days 1, 7, and 14, a subset of mice from each group were harvested, and their brains were collected after anesthesia. The brain tissue was stained in a 2% 2,3,5-triphenyltetrazolium chloride saline solution. Normal tissue stained red, and infarcted areas stained white. The percentage of infarct volume relative to the contralateral hemisphere was calculated using image analysis software. Figure 15 and 16 It is evident that CPZ inhibited the percentage of infarct volume on the injured side after stroke.

[0075] 1.4.3 Detection of ceruloplasmin expression (Western Blot)

[0076] On postoperative day 1, brain tissue from the ischemic side and contralateral cortex was harvested. Total protein was extracted, subjected to SDS-PAGE electrophoresis, transferred to a membrane, and incubated with anticeruloplasmin primary antibody and corresponding secondary antibody. CP protein expression levels were detected by chemiluminescence immunoassay. β-actin was used as an internal control.

[0077] 1.4.4 Observation of myelin regeneration and oligodendrocytes (immunofluorescence staining)

[0078] Brain tissue was harvested and frozen sections were prepared on postoperative days 1 and 7. Immunofluorescence double staining was performed using anti-CC1 antibody (to label mature oligodendrocytes and newly formed myelin) and Olig2 antibody (to label oligodendrocyte precursor cells). The number of CC1+ and Olig2+ cells was observed and counted under a confocal microscope in the corpus callosum and ischemic penumbra.

[0079] 1.4.5 Analysis of Iron Content and Distribution in the Brain

[0080] Brain tissue samples were harvested from the ischemic and control sides on postoperative days 1 and 7. Total iron content in one sample was analyzed using inductively coupled plasma mass spectrometry (ICP-MS). The other sample underwent MALDI mass spectrometry imaging to observe the spatial distribution of iron in the brain tissue.

[0081] 2. Results

[0082] 2.1 Neurological function recovery

[0083] Gait analysis showed that, compared with the model control group, mice in the CPZ treatment group exhibited significant improvement in motor function as early as day 1 post-surgery. Specifically, the stride length of the left forelimb, left hindlimb, right forelimb, and right hindlimb was significantly longer than that of the control group, closer to normal, and the differences were statistically significant. This improvement persisted until day 7 and day 14 post-surgery.

[0084] 2.2 Infarct volume

[0085] On postoperative day 1, TTC staining showed that the infarct volume in the CPZ treatment group was significantly smaller than that in the model control group. On postoperative day 7, the infarct volume in the CPZ treatment group continued to decrease, indicating that CPZ has a neuroprotective effect.

[0086] 2.3 Upregulation of ceruloplasmin expression

[0087] Western blot results showed that on postoperative days 1 and 7, the protein expression level of ceruloplasmin in the brain tissue (especially the ischemic side) of the CPZ treatment group was significantly higher than that in the model control group, indicating that CPZ effectively upregulated the expression of CP in the brain.

[0088] 2.4 Promote myelin regeneration

[0089] Immunofluorescence staining results showed that on postoperative day 7, the number of CC1+ mature oligodendrocytes in the ischemic penumbra and corpus callosum regions of the CPZ treatment group was significantly higher than that of the model control group. Simultaneously, the number of Olig2+ oligodendrocyte precursor cells also showed an increasing trend. This indicates that CPZ promotes the differentiation of OPCs into mature oligodendrocytes and myelin regeneration.

[0090] 2.5 Improves brain iron homeostasis

[0091] ICP-MS analysis showed that on postoperative day 1, the total iron content in the ischemic penumbra brain tissue of the model control group was significantly higher than that of the control side. The CPZ treatment group significantly promoted the balance of iron metabolism in the ischemic area and facilitated a faster recovery of iron content balance in both brain tissues. MALDI mass spectrometry imaging results visually demonstrated that the distribution of iron in the brain of the CPZ treatment group was more balanced, promoting iron recovery in the ischemic injury area.

[0092] This embodiment demonstrates that, in a mouse model of ischemic stroke, intravenous injection of dicyclohexanone oxalyl dihydrazone immediately after stroke can:

[0093] Significantly upregulated the expression of ceruloplasmin in the brain;

[0094] Effectively improves iron metabolism disorders in the brain;

[0095] Promotes oligodendrocyte precursor cell differentiation and myelin regeneration;

[0096] It significantly improves neurological deficits and reduces infarct volume in the very early post-stroke period (within 24 hours).

[0097] Figure 1 The image shows immunofluorescence staining of ceruloplasmin, which reveals that bicyclohexanone oxaloyl dihydrazone (CPZ) promotes CP expression in the corpus callosum. Figure 2 This is a protein immunoblotting map. Figure 3 for Figure 2 Protein expression statistics, Figure 2 and Figure 3 It is evident that CPZ promotes the balance of iron-related proteins in the control (CS) and damaged (IS) sides; Figure 4 Mass spectrometry imaging of iron in the brain of stroke mice after administration of CPZ shows that CPZ promotes rapid Fe homeostasis in the brain 1 day and 7 days after stroke. Figure 5 The total content of metal elements Cu, Zn, and Fe on the damaged side and the control side was determined by ICP-MS. Figure 6 Immunofluorescence staining of oligodendrocyte precursor cells in control mice one day after stroke; oligodendrocyte transcription factor 2 (O2), Dapi nuclear markers, Merge fitting plot; CS control side, IS lesion side; Figure 7 Immunofluorescence staining of oligodendrocyte precursor cells in mice treated with the stroke drug for one day; oligodendrocyte transcription factor 2, Dapi nuclear markers, Merge fitting plot; CS control side, IS lesion side; Figure 8 Immunofluorescence staining of oligodendrocyte precursor cells in control mice seven days after stroke; oligodendrocyte transcription factor 2 (O2), Dapi nuclear markers, Merge fitting plot; CS control side, IS lesion side; Figure 9 Immunofluorescence staining of oligodendrocyte precursor cells in mice treated with the drug seven days after stroke; oligodendrocyte transcription factor 2, Dapi nuclear markers, Merge fitting plot; CS control side, IS lesion side; Figure 10 for Figure 6-9 Fluorescence intensity statistics, from Figure 6-10 Immunofluorescence co-labeling showed that CPZ promoted the increase and differentiation of oligodendrocyte progenitor cells (OPCs) (Olig2+); Figure 11 Image of mature oligodendrocytes and ferritin light chains stained with immunofluorescence. Figure 12 for Figure 11The relative number of CC1-positive cells and fluorescence intensity statistics are shown in the graph, where Con 1dm stroke 1-day control group, CPZ 1dm stroke 1-day treatment group, Con 7dm stroke 7-day control group, CPZ 7dm stroke 7-day treatment group, Cs control side, Is lesion side, **p < 0.01, ***p < 0.001. Figure 13 for Figure 11 The relative number of FTL-positive cells and fluorescence intensity statistics are shown in the graph, where Con 1dm stroke 1-day control group, CPZ 1dm stroke 1-day treatment group, Con 7dm stroke 7-day control group, CPZ 7dm stroke 7-day treatment group, Cs control side, Is lesion side, **p < 0.01, ***p < 0.001. Figure 11-13 It is evident that CPZ promotes increased myelin sheath (CC1+) in the stroke injury area; Figure 14 The diagram shows the behavioral scoring of gait analysis. It can be seen that the functional improvement of the left forelimb (LF), left hindlimb (LH), right forelimb (RF), and right hindlimb (RH) in the CPZ group was significantly better than that in the control group. Figure 15 This is a schematic diagram of the TTC staining results; Figure 16 for Figure 15 TTC-stained lesion volume statistics, from Figure 15 and Figure 16 It is evident that 1 dm CPZ significantly reduces the damage volume.

[0098] Example 2: Injectable pharmaceutical formulation containing dicyclohexanone oxalyl dihydrazone

[0099] This embodiment discloses a lyophilized powder injection for intravenous injection, comprising:

[0100] Bicyclohexanone oxaloyl dihydrazone: 10 mg

[0101] Component 2: Mannitol: 50 mg

[0102] Citric acid (pH adjuster): Appropriate amount (to adjust pH to 6.0-7.5)

[0103] Water for injection: Add to 1.0 mL (when dissolved)

[0104] Preparation method:

[0105] (1) Under sterile conditions, dissolve the prescribed amount of dicyclohexanone oxaloyl dihydrazone and mannitol in an appropriate amount of water for injection and stir until completely dissolved;

[0106] (2) Adjust the pH to 6.0-7.5 using 0.1 mol / L citric acid solution;

[0107] (3) Make up the volume to 1.0 mL with water for injection and filter sterilely using a 0.22 μm microporous membrane;

[0108] (4) Dispense the filtrate into vials under aseptic conditions, 1.0 mL per vial;

[0109] (5) Freeze-dry the dispensed vials, then stopper and cap them to obtain the final product.

[0110] Instructions for use: Dissolve in 5 mL of 0.9% sodium chloride injection or 5% glucose injection before use, and administer slowly intravenously. The daily dose is 10-20 mg / kg, calculated based on the patient's weight.

[0111] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. The use of a dicyclohexanone oxalyl dihydrazone in the preparation of stroke drugs.

2. The use according to claim 1, characterized in that, The stroke mentioned is ischemic stroke.

3. The use according to claim 1, characterized in that, The drug is administered within 24 hours of the stroke.

4. The use according to claim 1, characterized in that, The effective dosage of the bicyclohexanone oxaloyl dihydrazone is 10-20 mg / kg per day.

5. The use according to claim 1, characterized in that, The bicyclohexanone oxaloyl dihydrazone achieves its use through at least one of the following mechanisms: a) upregulating the expression of ceruloplasmin in the brain; b) Enhances the activity of ferrooxidase in ceruloplasmin in the brain; c) Promotes the transport or reuse of iron in the brain; d) Promotes the survival, differentiation, or myelination of oligodendrocyte precursor cells; e) Promotes the repair of neurological function after stroke.

6. A stroke drug, characterized in that, This includes dicyclohexanone oxalyl dihydrazone, or its pharmaceutically acceptable salts, esters, or prodrugs.

7. The stroke medication according to claim 6, characterized in that, The stroke mentioned is ischemic stroke.

8. The stroke medication according to claim 6, characterized in that, The stroke medication is administered via intravenous injection.

9. The stroke medication according to claim 6, characterized in that, The stroke medication is administered within 24 hours of the stroke.

10. The stroke medication according to claim 6, characterized in that, The stroke medication also includes pharmaceutically acceptable carriers, excipients, or diluents.

Images