Use of compound inf195 in the treatment of diseases associated with nerve axon injury

Abstract

The application discloses application of a compound INF195 in treatment of diseases related to nerve axon injury. 17 H 22 ClNO3.The application proves through research that the compound INF195 can significantly promote axon regeneration of DRG neurons at a concentration of 5 muM, and has no significant effect on neuron activity at the concentration.Further based on an axon injury model of Xona microfluidic chip, it is proved that the INF195 can promote regeneration and extension of axons after injury.Therefore, the compound INF195 can inhibit NLRP3 inflammasome activity, and is an important compound molecule for promoting axon regeneration and repair after nerve injury, and has a good application prospect in preparation of drugs for treating diseases related to nerve axon injury, including peripheral nerve injury and central nerve injury.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to the application of compound INF195 in the treatment of diseases related to nerve axon injury. Background Technology

[0002] Nerve injury, particularly axonal rupture, loss, or long-distance defect, is a common neurological disorder in clinical practice, caused by various factors such as trauma, ischemia, metabolic diseases, and nerve compression. Nerve injury can lead to sensory disturbances, motor dysfunction, and even permanent disability, placing a heavy burden on patients' families and the social healthcare system. Depending on the location of the injury, nerve injury is mainly divided into peripheral nerve injury (PNI) and central nerve injury (CNI). While peripheral nerve injury possesses a certain degree of spontaneous regeneration capacity, the regeneration rate is slow and functional recovery is incomplete; whereas after central nerve injury, due to the presence of an inhibitory microenvironment, axons can hardly regenerate spontaneously, resulting in a very poor prognosis.

[0003] Currently, clinical treatment strategies for nerve injury mainly include autologous nerve transplantation, allogeneic nerve transplantation, nerve conduit repair, and local application of neurotrophic factors. However, these methods all have significant limitations: autologous nerve sources are limited and can cause secondary damage; allogeneic nerves pose a risk of immune rejection; the repair effects of tissue-engineered materials are unstable; and neurotrophic factors have short half-lives and are difficult to deliver precisely. Therefore, developing small molecule compound drugs that can effectively promote nerve axon regeneration has become a research hotspot and an urgent clinical need in the field of nerve injury repair.

[0004] In recent years, the crucial role of neuroinflammation in the repair of nerve injuries has received widespread attention. Studies have shown that after nerve injury, the NOD-like receptor family pyrin domain-containing 3 (NLRP3) inflammasome is abnormally activated in the injured area and the dorsal root ganglion (DRG), thereby inducing the maturation and release of pro-inflammatory factors such as interleukin-1β (IL-1β) and interleukin-18 (IL-18), forming a persistent inflammatory microenvironment that severely inhibits axonal regeneration and functional recovery. Therefore, targeting and inhibiting NLRP3 inflammasome activity has become a potential new strategy to promote axonal regeneration after nerve injury.

[0005] INF195 (CAS No.: 1211379-56-4, Molecular Formula: C) 17 H 22ClNO3 is a novel small molecule compound, and studies have confirmed its protective effect as an NLRP3 inhibitor in a myocardial ischemia-reperfusion injury model. However, no research has been reported to date regarding the application of INF195 in the nervous system, particularly its role in promoting axonal regeneration. Summary of the Invention

[0006] To address the shortcomings of existing technologies, this invention provides the application of compound INF195 in the treatment of diseases related to nerve axon injury. It is the first discovery that compound INF195 significantly promotes axon regeneration and axon repair after injury in DRG neurons at non-cytotoxic concentrations by inhibiting NLRP3 inflammasome activity, providing a novel candidate drug and application strategy for the treatment of diseases related to nerve axon injury.

[0007] This invention is achieved through the following technical solution:

[0008] The use of compound INF195, its pharmaceutically acceptable salt, or its solvates in the preparation of medicaments for treating diseases related to nerve axon injury, wherein the chemical structure of compound INF195 is shown in formula (I):

[0009]

[0010] Formula (I).

[0011] Preferably, the nerve axon injury-related diseases include peripheral nerve injury or central nerve injury.

[0012] Preferably, the compound INF195 promotes axonal regeneration at a concentration of 5 μM.

[0013] Preferably, the compound INF195 promotes the regeneration of nerve axons after injury by inhibiting the activity of the NLRP3 inflammasome.

[0014] Preferably, the compound INF195 can promote the regeneration and extension of axons after injury in the Xona microfluidic chip axon injury model.

[0015] A pharmaceutical composition for promoting the repair of nerve axonal injury includes a therapeutically effective amount of compound INF195 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient; the chemical structure of said compound INF195 is shown in formula (I).

[0016] Preferably, the excipients include one or more of the following: diluent, excipient, filler, binder, wetting agent, disintegrant, absorption promoter, surfactant, adsorbent carrier, and lubricant.

[0017] Preferably, the pharmaceutical composition is an oral dosage form or a non-oral dosage form.

[0018] The beneficial effects of this invention are as follows:

[0019] (1) This invention reveals for the first time the novel application of NLRP3 inflammasome inhibitor INF195 in promoting nerve axon regeneration. In the prior art, INF195 has only been reported for its protective effect against myocardial ischemia-reperfusion injury, and its application in the field of nervous system diseases, especially in nerve injury repair, has not been reported. Therefore, this invention fills the research gap of INF195 in the field of nerve regeneration and opens up a new pharmaceutical direction for it.

[0020] (2) This invention uses a primary DRG neuron culture model to verify the effective concentration of INF195 in promoting axon regeneration and its effect. The experimental results show that INF195 has no obvious cytotoxic effect on DRG neurons in the concentration range of 0~10 μM and no significant change in cell activity. However, at a concentration of 5 μM, the total axon length and the longest axon length of neurons in the INF195 treatment group are significantly increased compared with the control group, indicating that INF195 can effectively promote axon extension at this concentration. At the same time, it is clear that 5 μM is the optimal effective concentration, providing a precise dosage reference for subsequent formulation development.

[0021] (3) The present invention uses Xona microfluidic chip to construct an axon injury model to simulate the pathological microenvironment after nerve injury in vivo, and further verify the promoting effect of INF195 on axon regeneration after injury. The results show that the total length of regenerated axons and the longest axon length in the axon cavity of the INF195 treatment group are significantly higher than those of the control group. This model can realize the spatial separation of axons and cell bodies, more realistically reflect the axon regeneration process after injury, and enhance the reliability of experimental data and clinical translation value.

[0022] (4) This invention clarifies that INF195 promotes axon regeneration by inhibiting the activity of NLRP3 inflammasomes and reducing the inflammatory microenvironment after nerve injury. The elucidation of this mechanism provides a new therapeutic target for nerve injury repair and lays a theoretical foundation for the application of NLRP3 inhibitors in nervous system diseases. It has important academic value and clinical guiding significance.

[0023] (5) This invention provides a new candidate compound molecule for nerve injury repair. INF195 has a well-defined structure and can be used as a lead compound for structural optimization to develop more active nerve repair drugs. It can be used in combination with neurotrophic factors, tissue engineering materials, etc. to exert a synergistic repair effect. In the future, it can be developed into various dosage forms such as injections, local administration preparations, and sustained-release preparations to meet different clinical needs. It is suitable for various nerve injury populations such as patients with peripheral nerve injury, spinal cord injury, and traumatic brain injury. Therefore, the compound INF195 provided by this invention has broad application prospects in the preparation of drugs for the treatment of nerve axon injury-related diseases and has important clinical translational value and market development potential. Attached Figure Description

[0024] Figure 1 This is a comparison of the cytotoxicity of different concentrations of compound INF195 on DRG neurons in Example 2;

[0025] Figure 2 The effect of different concentrations of compound INF195 on axonal regeneration of DRG neurons in Example 3: A is a representative image of β-Tubulin III immunofluorescence staining (scale bar = 20 μm); B is a statistical graph of the number of neuronal axons; C is a statistical graph of the total length of neuronal axons; D is a statistical graph of the length of the longest neuronal axon.

[0026] Figure 3 To verify the effect of INF195 on axonal regeneration after injury using the Xona microfluidic chip in Example 4: A is an immunofluorescence staining image of axonal regeneration in the axonal cavity of different treatment groups (scale bar = 50 μm); B is a statistical graph of the total length of regenerated axons; C is a statistical graph of the longest axonal length.

[0027] Figure 4 Example 5 shows the effect of INF195 on NLRP3 pathway protein expression in DRG neurons as detected by Western blot: A is the Western blot result; B is the quantitative statistical graph of protein band gray values.

[0028] Figure 5 The effect of compound INF195 at a concentration of 5 μM in Example 6 on axonal regeneration after sciatic nerve injury in rats: A is a representative image of rat sciatic nerve axons stained with SCG10 (scale bar = 1000 mm); B is a quantitative statistical graph of the longest length of sciatic nerve axons. Detailed Implementation

[0029] The present invention will now be described in further detail with reference to the accompanying drawings and specific embodiments.

[0030] Unless otherwise specified, the technical means used in the following embodiments are all conventional means well known to those skilled in the art, and the experimental methods without specific conditions are all conventional methods in the art.

[0031] Unless otherwise specified, all materials and reagents used in the following examples are commercially available.

[0032] In the quantitative experiments involved in the following examples, at least three replicate experiments were set up, and the average value of the results was taken.

[0033] The experimental animals used in the following examples were SD rats, which were purchased from the Experimental Animal Center of Nantong University.

[0034] Example 1: Obtaining compound INF195

[0035] Compound INF195, CAS number 1211379-56-4, molecular formula C 17 H 22 ClNO3 has a molecular weight of 323.81. Its chemical structural formula is shown in formula (I):

[0036]

[0037] Formula (I)

[0038] The compound INF195 used in this example was purchased from MedChemExpress (MCE), catalog number HY-164304. The compound has a purity of ≥99% and can be used directly in cell experiments.

[0039] Example 2: Detection of the cytotoxic effect of compound INF195 on DRG neurons

[0040] 1. Isolation and culture of rat DRG neurons

[0041] Newborn SD rats (within 24 hours) were used. The spine was aseptically dissected, and all dorsal root segment tissues were removed using micro-forceps under a dissecting microscope and placed in pre-chilled dissection solution containing antibiotics. After all tissues were removed, the dissection solution was discarded, and the tissues were rinsed 2-3 times with sterile PBS. 2 mL of pre-warmed collagenase (3 mg / mL) was added to the tissues, and the tissues were thoroughly minced using microscissors and transferred to centrifuge tubes. The tubes were then incubated in a cell culture incubator for 90 min, with gentle agitation every 30 min. After digestion, the tubes were centrifuged at 800 rpm for 5 min, and the supernatant was discarded. 1 mL of pre-warmed trypsin (0.25%) was added to the pellet, and the pellet was gently resuspended by pipetting. The tubes were then incubated for another 10 min, with pipetting every 5 min. When the tissues became flocculent and free of obvious clumps, 3 mL of culture medium containing 10% FBS was added to terminate the digestion. The cells were dispersed by repeated pipetting with a 1 mL pipette tip and filtered through a 70 μm cell sieve into a new centrifuge tube. Centrifuge the filtrate at 1200 rpm for 5 min and discard the supernatant. Resuspend the cells in 4 mL of preheated 10% BSA solution, centrifuge at 900 rpm for 5 min, and carefully aspirate any floating myelin fragments and other cells. Repeat this step once. Resuspend the cells in an appropriate amount of preheated neuronal culture medium (Neurobasal + B27), count the cells, adjust the density, and seed them in 96-well plates pre-coated with poly-L-lysine (1×10⁻⁶ cells / well). 4 Cells / well were incubated in a 37°C, 5% CO2 incubator.

[0042] 2. Cytotoxicity detection

[0043] DRG neurons cultured for 24 h were divided into 5 groups, with 4 replicates per group. Compound INF195 was added to each well at final concentrations of 0, 1, 2, 5, and 10 μM, respectively. The control group received an equal volume of DMSO. After another 24 h of culture, 10 μL of CCK-8 assay reagent was added to each well, gently vortexed, and incubated for 2 h. The absorbance at 450 nm was measured using a microplate reader, and cell viability was calculated.

[0044] 3. Experimental Results and Analysis

[0045] like Figure 1 As shown, compared with the control group, after treatment with INF195 at concentrations of 0–10 μM for 24 h, the activity of DRG neurons did not change significantly (cell survival rate was >95%), indicating that INF195 had no significant cytotoxic effect on neurons within this concentration range.

[0046] Example 3: Study on the effect of compound INF195 on axonal regeneration of DRG neurons

[0047] 1. Isolation and culture of rat DRG neurons

[0048] The operating steps are the same as in Example 2.

[0049] 2. Neuron inoculation and drug treatment

[0050] The isolated DRG neurons were seeded at an appropriate density onto cell spreaders pre-coated with poly-L-lysine in 24-well plates and cultured in an incubator. After 48 h of culture, the culture medium was replaced with fresh medium, and different concentrations of compound INF195 (0, 1, 2, 5, 10 μM) were added. The control group was treated with an equal volume of DMSO. Immunofluorescence staining was performed after another 24 h of culture.

[0051] 3. Immunofluorescence staining

[0052] Discard the culture medium and gently wash the cells twice with pre-warmed PBS. Add pre-cooled 4% paraformaldehyde and fix at room temperature for 20 min. Discard the fixative and wash the cells three times with PBS for 5 min each time. Add blocking buffer containing 5% BSA and block at room temperature for 1 h. Discard the blocking buffer and add Anti-β-Tubulin III antibody (1:500, Abcam) diluted with primary antibody dilution buffer and incubate overnight at 4°C. Recover the primary antibody and wash the cells three times with PBS for 5 min each time. Add Alexa Fluor 594-labeled goat anti-mouse IgG (1:1000, Invitrogen) diluted with secondary antibody dilution buffer and incubate at room temperature in the dark for 1 h. Discard the secondary antibody and wash the cells three times with PBS for 5 min each time. Mount the cells with anti-fluorescence quenching mounting medium, observe and photograph them under a fluorescence microscope. Use ImageJ software to count the number of axons, total axon length, and longest axon length of each group of neurons. At least 80 neurons were counted in each group.

[0053] 4. Experimental Results and Analysis

[0054] like Figure 2 As shown, compared with the control group, at a concentration of 5 μM INF195, the number of neuronal axonal branches ( Figure 2 (B) Total length of neuronal axons ( Figure 2 (C) and longest axon length ( Figure 2 Both D and D levels increased significantly ( ** (P < 0.01), indicating that 5 μM INF195 can effectively promote axonal regeneration in DRG neurons. Specific statistical results are shown in Table 1.

[0055] Table 1. Effects of different concentrations of INF195 on axonal regeneration of DRG neurons

[0056]

[0057] Example 4: Study on the effect of INF195 based on Xona microfluidic chip on axonal regeneration after injury

[0058] The Xona microfluidic chip used in this embodiment was purchased from Xona Microfluidics. ® The company's chip structure comprises two independent cell culture chambers (cell body chamber and axon chamber) connected by an array of microchannels (450 μm in length, 10 μm in width, and 3 μm in height). This microchannel size design allows only axonal extension through, while the neuronal cell body is confined within the cell body chamber, thereby enabling directed axonal growth and independent manipulation.

[0059] 1. Microfluidic chip preparation

[0060] Place the Xona microfluidic chip in a sterile culture dish and sterilize it by UV irradiation for 30 min before use. Slowly inject 0.1 mg / mL poly-L-lysine solution into the channels, ensuring the liquid fills all microchannels, and coat overnight at 4°C. Before use, aspirate the poly-L-lysine, slowly rinse the chip three times with sterile PBS for 10 min each time, and finally add neuronal culture medium (Neurobasal + B27) to both chambers of the chip, and incubate at 37°C for 2 h to equilibrate.

[0061] 2. DRG neuron seeding and axonal directed culture

[0062] DRG neurons from newborn SD rats were harvested according to the method described in Example 2, and the cell density was adjusted to 1×10⁻⁶. 6 Cells / mL, slowly inject 10 μL of cell suspension into the Xona chip's cell lumen seeding wells (approximately 1 × 10⁶ cells per chip). 4 (Number of cells). The chip was placed in a 10 cm culture dish and incubated at 37°C with 5% CO2 for 2 hours to allow the cells to adhere fully. After cell adhesion, complete neuronal culture medium (Neurobasal + B27) was added to the cell body lumen and axonal lumen, respectively. During the operation, the fluid level in the cell body lumen was kept slightly higher than that in the axonal lumen, using the hydrostatic pressure difference to guide the directional growth of axons. Axonal growth was observed daily, and half of the culture medium was replaced every 2 days. By days 5-7, a large number of axons could be seen extending into the axonal lumen through the microchannels.

[0063] 3. Establishment of axonal injury model

[0064] Following the standard axonal injury modeling method for Xona chips, axonal injury was induced by a combination of physical traction and fluid shear force. On day 7 of culture, after confirming under a microscope that the axons had fully grown into the axonal cavity, the culture medium in the axonal cavity was removed by vacuum pump at a constant flow rate for 5 seconds, causing physical detachment of the axons. Preheated neuronal culture medium was immediately added to the axonal cavity, and preheated neuronal culture medium with different treatments was added to the cell body cavity. The chip was then transferred to an incubator for further culture.

[0065] 4. Drug treatment grouping

[0066] The Xona chips that successfully established axonal injury models were randomly divided into the following two groups:

[0067] ① Control group: Neuronal culture medium containing DMSO (1:1000) was added to the cell cavity;

[0068] ②INF195 treatment group: Neuronal culture medium containing 5 μM INF195 was added to the cell cavity.

[0069] Each group was cultured for another 48 hours, with the corresponding drug culture medium replaced every 24 hours during this period, and the liquid level difference between the two chambers was maintained.

[0070] 5. Immunofluorescence detection of axonal regeneration after injury

[0071] After culture, the culture medium was discarded, and pre-cooled 4% paraformaldehyde was slowly injected into the Xona chip channels, and the chip was fixed at room temperature for 30 min. The chip was slowly rinsed three times with PBS for 10 min each time. Blocking solution containing 5% BSA was injected into the chip channels, and the chip was blocked at room temperature for 1 h. Anti-β-Tubulin III antibody (1:500, Abcam) diluted with primary antibody dilution buffer was injected, and the chip was incubated overnight at 4°C. The next day, the chip was rinsed three times with PBS for 10 min each time. Secondary antibody labeled with Alexa fluor 488 (1:1000, Invitrogen) was injected, and the chip was incubated at room temperature in the dark for 2 h. After rinsing with PBS, anti-fluorescence quenching mounting medium was injected. Axonal regeneration in the axonal cavity was observed and photographed under a confocal microscope. The total length and longest axon length of regenerated axons in the axonal cavity of each group were counted based on the damage treatment time.

[0072] 6. Experimental Results and Analysis

[0073] like Figure 3 As shown, the total length of regenerated axons in the axonal cavity of the INF195 treatment group ( Figure 3 (B) and the longest axon length ( Figure 3 The C values ​​in the middle were significantly higher than those in the control group (C). ** P < 0.01, indicating that INF195 can effectively promote regeneration and repair after axonal injury.

[0074] Example 5: Western Blot analysis of the effect of INF195 on NLRP3 pathway protein expression in DRG neurons.

[0075] 1. Drug treatment

[0076] To verify the inhibitory effect of compound INF195 on NLRP3 inflammasome activation in DRG neurons, primary cultured DRG neurons (isolation and culture method as in Example 2) were seeded in 6-well plates and cultured for 48 h, after which the culture medium was replaced with fresh medium. The cells were divided into two groups: a control group treated with an equal volume of DMSO, and an INF195 treatment group treated with a final concentration of 5 μM INF195. After culturing for another 24 h, total protein was extracted from both groups of cells for subsequent Western blot analysis.

[0077] 2. Western Blot Detection and Analysis

[0078] The total protein extracted from DRG neurons was concentrated using the BCA method. 25 μg of total protein was then separated by SDS-PAGE electrophoresis and transferred to a PVDF membrane, which was blocked with 5% skim milk at room temperature for 2 h. The following primary antibodies were added: NLRP3 (1:1000 dilution, Immunoway), ASC (1:1000 dilution, Abcam), Caspase-1 (1:1000 dilution, Abcam), and IL-1β (1:10000 dilution, Proteintech), and incubated overnight at 4°C. The next day, the corresponding HRP-labeled secondary antibodies (Jackson) were added, and the cells were incubated at room temperature for 2 h. Development was performed using ECL chemiluminescence. GAPDH was used as an internal control protein to detect the expression levels of NLRP3 pathway-related proteins in each group of cells. The experiment was repeated three times, and grayscale analysis was performed using ImageJ software.

[0079] 3. Experimental Results and Analysis

[0080] The results are as follows Figure 4 As shown, compared with the control group, the expression level of NLRP3 protein in the 5 μM INF195 treatment group did not change significantly, but the expression levels of ASC, Caspase-1 and IL-1β proteins were significantly reduced.

[0081] The experimental results of this embodiment demonstrate that INF195 does not reduce the expression of downstream ASC aptamers and the maturation and release of Caspase-1 and IL-1β by inhibiting the assembly or activation of the NLRP3 inflammasome, rather than by suppressing the protein expression level of NLRP3. This ultimately alleviates the inflammatory response in neurons. These results further clarify the mechanism by which INF195 inhibits NLRP3 inflammasome activation in DRG neurons.

[0082] Example 6: Effect of compound INF195 on axonal regeneration after sciatic nerve injury in rats

[0083] 1. Establishment of a rat model of sciatic nerve pinching injury and administration of drugs within the DRG (Diagnosis Related Groups).

[0084] Eight-week-old male SD rats (weighing 180 g) were anesthetized by intraperitoneal injection of sodium pentobarbital (40 mg / kg). The iliac region of the back and hind limbs were located and the skin was prepared and disinfected. The skin was incised, the muscle tissue was bluntly dissected, the corresponding spinous processes were exposed and removed, and the L4 and L5 dorsal root ganglia (DRGs) were exposed. At the same time, the sciatic nerve was isolated and exposed. A sciatic nerve clamping model was established by clamping the proximal end of the sciatic nerve with a clamping width of 3 mm for 30 seconds.

[0085] After the pinching injury was completed, the rats were randomly divided into two groups (n=4 per group): the control group received a slow injection of 2 μL of solvent control (DMSO) into the DRG using a microsyringe, and the INF195 treatment group received an injection of 2 μL of 5 μM INF195 solution into the DRG. After the injection, the nerve was repositioned, and the muscles and skin were sutured. Postoperative care included keeping the rats warm and administering anti-infection treatment.

[0086] 2. Tissue irrigation fixation and dehydration treatment

[0087] On the third day after sciatic nerve injury, sciatic nerve tissue was harvested for examination. After anesthetizing the rats, the abdominal cavity was opened to expose the heart. A perfusion needle was inserted through the apex of the heart into the aorta, and physiological saline was rapidly infused first. Simultaneously, the right atrial appendage was cut open, and after the blood was drained, the perfusion was replaced with pre-cooled 4% paraformaldehyde solution until the rats' limbs became rigid. The sciatic nerve was isolated and fixed in 4% paraformaldehyde solution at 4°C for 8 hours. After washing with PBS, it was transferred to 30% sucrose solution for dehydration at 4°C. Once the tissue had settled, it was ready for embedding.

[0088] 3. Sciatic nerve frozen section and immunofluorescence staining

[0089] The dehydrated sciatic nerve was removed, excess connective tissue was trimmed, and the tissue was rapidly frozen with OCT embedding medium and stored at -20°C. Longitudinal sections were prepared using a cryostat, with a section thickness of 18 μm. The sections were attached to poly-L-lysine-coated slides and dried at 37°C. Before staining, the sections were warmed to room temperature, washed three times with PBS, and the tissue was circled with an immunohistochemical pen. Blocking buffer was added and the sections were blocked at room temperature for 1 h. The blocking buffer was discarded, and primary antibody SCG10 (1:400 dilution) was added and incubated overnight at 4°C. The next day, after warming to room temperature, the sections were washed three times with PBS, and Alexa Fluor 594-labeled fluorescent secondary antibody (1:1000 dilution) was added in the dark and incubated at room temperature for 2 h. After washing again in the dark, the sections were mounted with anti-fluorescence quenching mounting medium containing DAPI and observed and photographed under a fluorescence microscope. The longest length of the regenerated axon was measured using ImageJ software.

[0090] 4. Experimental Results and Analysis

[0091] like Figure 5 Image A shows a representative image of sciatic nerve axon SCG10 immunofluorescence staining (red indicates SCG10 positive regenerated axon), showing that after sciatic nerve injury, the length of regenerated axons in the axonal cavity of the INF195 treatment group was significantly longer than that of the control group. Figure 5 In the middle B, there is a quantitative statistical graph of the longest length of the regenerated axon.

[0092] The experimental results of this embodiment show that after sciatic nerve injury, treatment with 5 μM INF195 can significantly promote the regeneration and extension of the injured axon. The significant increase in the longest length of the regenerated axon confirms that INF195 also has the effect of promoting nerve axon regeneration at the in vivo level.

[0093] The embodiments described above are only some, not all, of the embodiments of the present invention. The detailed description of the embodiments of the present invention is not intended to limit the scope of the claimed invention, but merely to illustrate selected embodiments. The scope of protection of the present invention is determined by the scope claimed in the claims. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without inventive effort are within the scope of protection of the present invention.

Claims

1. The use of compound INF195, its pharmaceutically acceptable salt, or its solvate in the preparation of a medicament for treating diseases related to nerve axon injury, characterized in that, The chemical structure of the compound INF195 is shown in formula (I): Formula (I).

2. The application according to claim 1, characterized in that, The nerve axon injury-related diseases include peripheral nerve injury or central nerve injury.

3. The application according to claim 1, characterized in that, The compound INF195 promotes axonal regeneration at a concentration of 5 μM.

4. The application according to claim 1, characterized in that, The compound INF195 promotes the regeneration of nerve axons after injury by inhibiting the activity of the NLRP3 inflammasome.

5. The application according to claim 1, characterized in that, The compound INF195 was able to promote the regeneration and extension of axons after injury in the Xona microfluidic chip axon injury model.

6. A pharmaceutical composition for promoting the repair of nerve axonal injury, characterized in that, It includes a therapeutically effective amount of the compound INF195 or a pharmaceutically acceptable salt thereof, and pharmaceutically acceptable excipients; the chemical structure of the compound INF195 is shown in formula (I).

7. The pharmaceutical composition according to claim 6, characterized in that, The excipients include one or more of the following: diluent, excipient, filler, binder, wetting agent, disintegrant, absorption promoter, surfactant, adsorbent carrier, and lubricant.

8. The pharmaceutical composition according to claim 6, characterized in that, The pharmaceutical composition may be in oral or non-oral form.

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