Pharmaceutical application of PLX5622

Abstract

The invention relates to the field of medicines, in particular to pharmaceutical application of PLX5622. The invention finds that PLX5622 or a pharmaceutically acceptable salt thereof can be used for preparing a medicine for preventing and treating retina structure damage and / or function damage caused by retinal ischemia reperfusion injury caused by retinal artery occlusion, and solves the problem that in the prior art, the vision of a patient with retinal artery occlusion is difficult to recover after blood is recirculated.

Description

Technical Field

[0001] The present invention relates to the field of medicine, and in particular to pharmaceutical use of PLX5622. Background Art

[0002] Retinal artery occlusion (RAO) is a serious ophthalmic emergency. Its etiology is mainly due to sudden occlusion of the central retinal artery, which leads to acute dysfunction of retinal tissue. Acute retinal artery occlusion causes axoplasmic stasis and intracellular edema, which usually leads to permanent retinal ischemia and irreversible death of retinal ganglion cells (RGCs) within a few hours. At present, the treatment of RAO in clinical practice is mainly aimed at the treatment of retinal occlusion emboli. The treatment methods include anterior chamber puncture, acetazolamide treatment, eye massage, and arteriovenous drugs or interventional surgery thrombolysis. The goal of treatment is to improve and restore retinal circulation and avoid retinal necrosis. However, the time window for RAO treatment is short, and after the retinal artery is restored to blood perfusion, the original function of the retina is not only not restored, but the structural damage is more serious, and even irreversible damage occurs. This phenomenon is called retinal ischemia-reperfusion injury (RIRI). It is reported that only 20% of patients can recover their vision to a limited extent after retreatment. At present, the pathogenesis of RAO and its ischemia-reperfusion injury has not been fully elucidated. How to maximally save the ischemic but not infarcted retina after RAO treatment, reduce further oxidative damage caused by RIRI, and restore the structure and function of the retina has become a difficult problem that needs to be solved urgently. Summary of the invention

[0003] The inventors have found that the use of PLX5622 after blood circulation is restored after retinal artery occlusion can prevent the occurrence of retinal structural damage and / or functional damage, solving the problem of difficulty in restoring vision after blood circulation is restored in patients with retinal artery occlusion in the prior art. Based on this, the present invention provides the use of PLX5622 or a pharmaceutically acceptable salt thereof in the preparation of a drug for preventing and treating retinal structural damage and / or functional damage caused by retinal ischemia-reperfusion injury caused by retinal artery occlusion.

[0004] The technical solution provided by the present invention is as follows: The present invention provides the use of PLX5622 or a pharmaceutically acceptable salt thereof in the preparation of a drug for preventing and treating retinal ischemia-reperfusion injury caused by retinal artery occlusion leading to retinal structural damage and / or functional damage.

[0005] In some embodiments of the present invention, the damaged retinal structure is caused by damage or death of retinal ganglion cells.

[0006] In some embodiments of the present invention, the drug is a drug for treating retinal artery occlusion or a drug for preventing complications after treatment of retinal artery occlusion.

[0007] In some embodiments of the present invention, the therapeutic drug is an intra-arterial thrombolytic drug, and PLX5622 or a pharmaceutically acceptable salt thereof is used as an auxiliary component in the intra-arterial thrombolytic drug, and the auxiliary component is used to prevent retinal structural damage and / or functional damage.

[0008] In some embodiments of the present invention, the arterial thrombolytic drug contains a fibrinolytic component and / or a vasodilator component as main components.

[0009] In some embodiments of the present invention, the treatment of retinal artery occlusion includes one or more of anterior chamber puncture, acetazolamide treatment, eyeball massage, arterial thrombolytic drug thrombolysis, and interventional surgical thrombolysis.

[0010] In some embodiments of the invention, the complication is damage to the retinal structure and / or damage to its function.

[0011] In some embodiments of the present invention, PLX5622 is in the form of a pharmaceutically acceptable acid addition salt.

[0012] In some embodiments of the present invention, the dosage form of the drug is one or more of an oral agent, a microcapsule preparation, an injection, a suppository, an eye drop, an eye wash, an eye cream, and an eye gel.

[0013] In some embodiments of the present invention, the symptom of structural damage is at least one of white lesions in the fundus and retinal edema; the symptom of functional damage is at least one of a sharp decrease in vision and visual field loss.

[0014] Compared with the prior art, the present invention has at least the following beneficial effects: In view of the problem of decreased vision after blood circulation is restored in retinal artery occlusion, there is a lack of effective therapeutic drugs in the prior art. The present invention provides the use of PLX5622 or a pharmaceutically acceptable salt thereof in the preparation of a drug for preventing and treating retinal ischemia-reperfusion injury caused by retinal artery occlusion, which leads to retinal structural damage and / or functional damage, thereby solving the problem in the prior art that it is difficult for patients with retinal artery occlusion to recover their vision after blood circulation is restored. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the accompanying drawings required for use in the description of the embodiments will be briefly introduced below. Obviously, the accompanying drawings in the following description are only some embodiments of the present invention. For ordinary technicians in this field, other accompanying drawings can be obtained based on these accompanying drawings without paying creative work.

[0016] Figure 1 The results of immunofluorescence staining of retinal ganglion cells in mice with pterygopalatine and ophthalmic artery occlusion model 7 days after low-dose (40 mg / kg / d) PLX5622 intervention; Con represents the healthy group, PLX5622 Con represents the PLX5622-administered healthy group, PLX5622 UPOAO represents the PLX5622-intervention group, and UPOAO represents the control group.

[0017] Figure 2 These are the results of retinal fundus photography and optical coherence tomography (OCT) of mice with artery occlusion model 7 days after low-dose (40 mg / kg / d) PLX5622 intervention, where: Con represents the healthy group, PLX5622 Con represents the PLX5622-administered healthy group, PLX5622 UPOAO represents the PLX5622-intervention group, and UPOAO represents the control group.

[0018] Figure 3 yes Figure 2 Statistical results of inner retinal thickness in OCT images.

[0019] Figure 4 The waveforms of the electroretinogram (ERG) a-wave and b-wave in mice with arterial occlusion model 7 days after low-dose (40 mg / kg / d) PLX5622 intervention are compared; wherein: Con represents the healthy group, PLX5622 Con represents the PLX5622-administered healthy group, PLX5622 UPOAO represents the PLX5622-intervention group, and UPOAO represents the control group.

[0020] Figure 5 There was no statistical difference in the retinal microglia count results of immunofluorescence mice in the artery occlusion model 7 days after low-dose (40 mg / kg / d) PLX5622 intervention; where: Con represents the healthy group, and PLX5622 Con represents the PLX5622-administered healthy group.

[0021] Figure 6 This is the result of CCK8 cell activity detection after BV2 cells were intervened by different concentrations of PLX5622 (0.1~5μM). DETAILED DESCRIPTION

[0022] In order to make the purpose, technical solution and advantages of the present invention clearer, the technical solution of the present invention will be clearly and completely described below in combination with the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0023] PLX5622 is a small molecule inhibitor that targets the colony stimulating factor 1 receptor (CSF1R). Existing technologies use PLX5622 to study the role of microglia in retinal protection. Studies have shown that PLX5622, as a CSF1R inhibitor, shows different effects in multiple retinal disease models by depleting microglia. In some cases, such as light-induced injury and retinal vein occlusion, PLX5622 can slow down retinal function degeneration and delay retinal degeneration. However, in glaucoma models, microglia depletion exacerbated the loss of retinal ganglion cells. These findings suggest that microglia have a complex dual role in retinal protection, which may protect retinal function or aggravate damage, depending on the disease model and experimental conditions.

[0024] The present invention used a low-dose (40 mg / kg / d) PLX5622 intervention on a mouse model after recanalization of retinal artery occlusion, and found that PLX5622 did not eliminate microglia, but had a protective effect on the retina of mice after recanalization of retinal artery occlusion, avoiding structural and / or functional damage to the retina caused by RIRI, which is different from the role of high-dose used to organize microglia clearance in previous reports.

[0025] The present invention provides the use of PLX5622 or a pharmaceutically acceptable salt thereof in the preparation of a drug for preventing and treating retinal ischemia-reperfusion injury caused by retinal artery occlusion, resulting in retinal structural damage and / or functional damage. In the present invention, a small amount of PLX5622 exhibits a significant protective effect on the retina of mice after blood circulation is restored after retinal artery occlusion. This discovery not only provides a new strategy for the treatment of retinal artery occlusion, but also reveals that the protective effect of PLX5622 on the retina is not achieved by completely eliminating microglia. This discovery provides a theoretical basis and experimental basis for the development of new drugs for retinal artery occlusion and its related complications, and is expected to bring better treatment options and prognosis to patients with retinal artery occlusion.

[0026] In some embodiments of the present invention, the retinal structural damage is caused by retinal ganglion cell damage or death. The present invention verifies that PLX5622 has a protective effect on retinal ganglion cells after retinal artery occlusion treatment, thereby protecting the structure and function of the retina from damage. The present invention uses technical means such as tissue section staining to observe in detail the survival status of retinal ganglion cells in different mice. The results showed that the RGCs cells stained with Rbpms in the retina of the control group (UPOAO) were significantly reduced compared with those of healthy mice (Con), indicating that RIRI caused RGCs cell death; while the RGCs cells stained with Rbpms in the retina of the PLX5622 intervention group (PLX5622 UPOAO) increased compared with the control group (UPOAO), indicating that PLX5622 effectively inhibited the apoptosis of RGCs cells during RIRI. In addition, the present invention found that the retinal function of the PLX5622 intervention group was significantly improved compared with the control group through the detection of electroretinogram, which was specifically manifested in that the amplitude and latency of the electrical signal were restored to a level close to that of healthy mice, further confirming the dual protective effect of PLX5622 on retinal structure and function. These findings not only deepen the understanding of the pharmacological mechanism of PLX5622, but also provide new perspectives and potential drug therapeutic targets for the clinical treatment of retinal artery occlusion.

[0027] In some embodiments of the present invention, the drug is a drug for treating retinal artery occlusion or a drug for preventing complications after treatment of retinal artery occlusion. PLX5622 can be used during the treatment of retinal artery occlusion or after treatment of retinal artery occlusion, and can protect the retina after blood supply is restored to the retinal artery occlusion to prevent damage to its structure and function.

[0028] In some embodiments of the present invention, the therapeutic drug is an arterial thrombolytic drug, and PLX5622 or a pharmaceutically acceptable salt thereof is used as an auxiliary component in the arterial thrombolytic drug, and the auxiliary component is used to prevent retinal structural damage and / or functional damage. The time window for RAO treatment is short, and using PLX5622 as an auxiliary component in the arterial thrombolytic drug can start to play a preventive role during the arterial thrombolysis process, avoiding retinal structural damage and / or functional damage caused by too late administration.

[0029] In some embodiments of the present invention, the arterial thrombolytic drug contains a fibrinolytic component and / or a vasodilator component as a main component. The arterial thrombolytic drug may use a fibrinolytic component and / or a vasodilator component as a main component and use PLX5622 as an auxiliary component.

[0030] In some embodiments of the present invention, the treatment of retinal artery occlusion includes one or more of anterior chamber puncture, acetazolamide treatment, eyeball massage, arterial thrombolytic drug thrombolysis, and interventional surgical thrombolysis. These treatment methods for retinal artery occlusion can restore blood supply to the retinal artery, but will induce RIRI. Immediately administering PLX5622 for prevention after the treatment of retinal artery occlusion can avoid structural and / or functional damage to the patient's retina and prevent the patient's vision from irreversibly declining.

[0031] In some embodiments of the present invention, the complication is retinal structural damage and / or functional damage. The present invention experimentally verified that the waveforms of the a-wave and b-wave of the electroretinogram (ERG) of mice with artery occlusion model did not change 7 days after low-dose (40 mg / kg / d) PLX5622 intervention, proving that intervention with PLX5622 after retinal artery occlusion treatment can avoid retinal functional damage.

[0032] In some embodiments of the present invention, PLX5622 is in the form of a pharmaceutically acceptable acid addition salt.

[0033] In some embodiments of the present invention, the dosage form of the drug is one or more of an oral agent, a microcapsule preparation, an injection, a suppository, an eye drop, an eye wash, an eye cream, and an eye gel. Optionally, the oral agent is one or more of a capsule, a tablet, a granule, and an oral solution; the injection is preferably an intraocular injection solution. The present invention confirms that a low dose of PLX5622 oral agent can prevent and treat retinal ischemia-reperfusion injury caused by retinal artery occlusion, resulting in retinal structural damage and / or functional damage. Compared with injections, suppositories, eye drops, eye washes, eye creams, eye gels and other preparations, PLX5622 oral agents are easy to use, have good patient compliance, and are safe.

[0034] In some embodiments of the present invention, the symptom of structural damage is at least one of white lesions in the fundus and retinal edema; the symptom of functional damage is at least one of a sharp decrease in vision and visual field loss. In an embodiment of the present invention, OCT images showed that there were white lesions in the fundus of mice in the control group after retinal ischemia, and no white lesions in the fundus were found in the mice in the PLX5622 intervention group, proving that PLX5622 intervention can prevent white lesions in the fundus. The fundus color photography results showed obvious ischemic edema around the optic disc of the mice in the control group, and no optic disc ischemic edema was found in the PLX5622 intervention group, proving that PLX5622 intervention can prevent retinal edema. ERG results showed that the b wave of the control group mice decreased significantly, indicating a sharp decrease in vision, and the b wave of ERG in the PLX5622 intervention group increased significantly, proving that PLX5622 intervention can significantly delay the decline in vision and play a role in protecting retinal function.

[0035] The technical solution provided by the present invention is described in detail below in conjunction with embodiments.

[0036] All experimental animals used, C57BL / 6 mice, were purchased from Liaoning Changsheng Biotechnology Co., Ltd. (certificate number: No. 210726231100785184).

[0037] The No. 602156 silicone plug (diameter 0.19±0.01mm, length 6.0±0.1mm) used was ordered from Pingdingshan Yushun Biotechnology Co., Ltd.

[0038] Drugs and reagents used: Isoflurane and oxygen / nitrous oxide mixed gas were purchased from Shenzhen Reward Technology Co., Ltd. Other conventional antibodies and reagents were conventional commercial products.

[0039] The instruments and equipment used were: the surgical microscope was Olympus SZX7 from Japan, the electroretinogram (ERG) was RetiMINER4.0 from Chongqing Ai Er Xi, and the fluorescence confocal microscope was TCS-SP8 from Germany. Others were common commercially available products.

[0040] Unless otherwise specified, the PLX5622 working solution and control solvent used in the examples were prepared by the following method and administered as follows: ① Preparation of PLX5622 mother solution: PLX5622 was dissolved in dimethyl sulfoxide (DMSO) to obtain a PLX5622 mother solution with a concentration of 100 mg / mL; ② Preparation of PLX5622 working solution: Polyethylene glycol 12-hydroxystearate (SolutolHS-15, TargetMol), polyethylene glycol 400 (PEG400, TargetMol) and double distilled water (dd water) were mixed into a dilution solvent at a volume ratio of 2:4:13. PLX5622 mother solution (100 mg / mL) was added to the dilution solvent at a volume ratio of 1:9 and mixed to obtain a PLX5622 working solution with a concentration of 5 mg / mL.

[0041] ③Preparation of control solvent: Mix DMSO and dilution solvent in a volume ratio of 1:19 to obtain a control solvent.

[0042] ④ The mice were weighed before daily gavage administration, and the dosage was adjusted according to the daily weight changes. The volume of PLX5622 working solution was calculated based on a daily dosage of 40 mg PLX5622 per kilogram of body weight.

[0043] Embodiment 1: (1) Establishment of the unilateral pterygopalatine artery occlusion (UPOAO) mouse model Experimental mouse preparation: Twenty healthy male C57BL / 6 mice aged 8-12 weeks and weighing 21g±1g were selected. The left eye of each mouse was the modeling group and the right eye was the control group. They were raised in an IVC system environment with a temperature of 24-17°C, a relative humidity of 60%, and free access to food and water for one week.

[0044] Silicone plug preparation: Prepare the thread plug in advance, with the specifications of silicone diameter 0.19±0.01mm, silicone length 6mm±0.1mm, and 0.4 fishing line. Put it in a 35mm sterile culture dish, soak it in 75% alcohol for 1 hour, and dry it naturally in a biosafety cabinet for later use.

[0045] Establishment of unilateral pterygopalatine artery occlusion (UPOAO) mouse model: S10. Anesthetize, fix, and control body temperature of C57BL / 6 mice: Use 2.0% isoflurane and oxygen / nitrous oxide mixed gas inhalation to anesthetize the mice. Put the mice in a supine position and fix them. Wet the hair on the neck with 75% medical alcohol, remove it with a blade, and wipe it clean. Insert a rectal temperature probe to maintain the body temperature at 37±0.5℃.

[0046] Separation of pterygopalatine artery: Expose the neck and separate the left common carotid artery, internal carotid artery and external carotid artery. Use 8-0 silk thread to ligate the distal end of the external carotid artery, insert another 8-0 silk thread into the external carotid artery, and tie a slipknot near the bifurcation of the common carotid artery. Make a midline incision in the neck to expose the left common carotid artery, internal carotid artery and external carotid artery. Use 8-0 silk thread to ligate the distal end of the external carotid artery, insert another 8-0 silk thread into the external carotid artery, and tie a slipknot near the bifurcation of the common carotid artery. Use artery clamps to clamp the internal carotid artery and common carotid artery respectively. Cut a small incision 1mm below the ligature of the external carotid artery.

[0047] S20. Insert No. 602156 silicone thread plug into the pterygopalatine artery to construct the model: insert a No. 602156 sterile silicone thread plug 1 mm below the external carotid artery ligature and gently push the thread plug with tweezers until all the silicone of the thread plug enters the pterygopalatine artery.

[0048] Model completion: Loosen the internal carotid artery clamp, cut the external carotid artery at the small opening, withdraw the thread plug, flip it to enter the internal carotid artery, and insert it into the pterygopalatine artery. The insertion depth of the thread plug is about 6±1mm from the bifurcation of the common carotid artery. The silicone tail of the thread plug is roughly located at the bifurcation of the common carotid artery, and the slipknot is slightly tightened. After 60 minutes of ischemia, the thread plug is pulled out, the proximal end of the external artery is ligated, and the artery clamp is loosened. Blood reperfusion of the common carotid artery and the internal carotid artery can be observed, and the wound is sutured to obtain the UPOAO model mouse.

[0049] (2) RAO treatment Take an appropriate amount of UPOAO model mice and randomly divide them into two groups: PLX5622 intervention group (abbreviated as PLX5622 UPOAO): The mice were fed with a regular diet and received continuous gavage of PLX5622 working solution every day for 7 days (from the first day to the seventh day), and the dosage was 40 mg PLX5622 per kilogram of body weight per day.

[0050] Control group (UPOAO): The mice were fed with a regular diet and received oral gavage of the same volume of control solvent as the UPOAO model mice in the PLX5622 intervention group.

[0051] At the same time, several healthy male C57BL / 6 mice of the same age were taken and divided into two groups: Healthy group (Con): The mice were fed with a regular diet and received intragastric administration of the same volume of control solvent as the mice in the PLX5622 intervention group.

[0052] PLX5622-administered healthy group (abbreviated as PLX5622 Con): The mice were fed with a regular diet and received continuous gavage of PLX5622 working solution for 7 days (from day 1 to day 7). The dosage was 40 mg PLX5622 per kilogram of body weight per day.

[0053] (3) Evaluation of the effectiveness of RAO treatment Three RAO-treated mice were selected from each group and general anesthetized by intraperitoneal injection of 2% sodium pentobarbital at a dose of 0.3 mL per 100 g body weight. Compound tropicamide mydriatic eye drops were dripped to dilate the pupil for 5 minutes. The dilated mice were placed on the small animal fundus imaging operating table, and fundus color photography and OCT acquisition of retinal structure were performed using the fundus and optical coherence tomography (OCT) modules. Figure 2 As shown in the figure, the control group mice had white lesions in the fundus after retinal ischemia, ischemic whitening edema appeared around the optic disc, and OCT images showed that the inner retinal structure was damaged. The mice in the PLX5622 intervention group had no white lesions in the fundus, no whitening edema appeared around the optic disc, and OCT images showed that the inner retinal structure was clearer and the retinal thickness increased (as shown in the box in the figure). Compared with the control group (UPOAO), the retinal structure of the PLX5622 intervention group (PLX5622 UPOAO) was intact. Figure 3The thickness of the inner retina in the OCT images was counted, and it was found that the thickness of the inner retina (nerve fiber layer and ganglion cell layer, RNFL+GCL) in the PLX5622 intervention group (PLX5622 UPOAO) was significantly increased compared with the control group (UPOAO) (*: P<0.05, **: P<0.01, ***: P<0.001, ****: P<0.0001), indicating that PLX5622 has a significant protective effect on the retinal structure of UPOAO model mice.

[0054] Eight RAO-treated mice were selected from each group and general anesthetized by intraperitoneal injection of 2% sodium pentobarbital at a dose of 0.3 mL per 100 g of body weight. After the mice were anesthetized stably, compound tropicamide mydriatic eye drops were dripped to dilate the pupil for 15 minutes. The dilated mice were placed on the electroretinogram (ERG) operating table with the head in an upright position and both eyes at the same height and fully exposed. The tail electrode clamped the rear end of the mouse tail and the subcutaneous electrode was inserted into the skin of the back of the neck. The left and right corneal contact electrodes were placed in contact with the model eye and the normal eye, and the amplitude changes of the a wave and b wave of the electroretinogram were recorded by stimulating the ERG with different light intensities. Figure 4 The waveforms of the a and b waves of the electroretinogram of the PLX5622-intervention group and the control group are shown. Comparison between the PLX5622-intervention group and the control group showed that, under different stimulation intensities, there was no statistical difference in the a wave, but the b wave was significantly increased (*: P<0.05, **: P<0.01, ***: P<0.001, ****: P<0.0001), indicating that low-dose PLX5622 has a protective effect on the visual function of UPOAO model mice.

[0055] To further verify the protective effect of PLX5622 on retinal ganglion cells (RGCs), retinal samples from mice in the PLX5622 intervention group and the control group were taken and fixed with 4% paraformaldehyde for 45 minutes. The retina was separated and dissected, and the retina was embedded to make retinal slices. The retinal slices were rinsed with PBS, and the retina was immersed in 5% BSAT solution (i.e., a mixture of 0.3% Triton X-100 in immunofluorescence blocking solution (BSA)), and placed in a 4°C refrigerator for blocking overnight. Rbpms sheep serum primary antibody was incubated in a 4°C refrigerator for 48 hours, rinsed in PBS buffer, incubated with AlexaFluor488 donkey anti-sheep fluorescent secondary antibody at 4°C overnight, rinsed in PBS buffer, and sealed with anti-fluorescence quenching sealing solution. The RGCs were counted using a confocal microscope. The results are shown in Figure 1As shown, compared with the healthy group, the number of RGCs cells stained with Rbpms on the retinal sections of the control group mice was significantly reduced, while the number of RGCs cells stained with Rbpms on the retinal sections of the PLX5622 intervention group mice was significantly increased compared with the control group, confirming that PLX5622 has a protective effect on the RGCs cells of UPOAO model mice.

[0056] (4) The neuroprotective effect of PLX5622 intervention on RAO is not achieved by clearing microglia (4.1) To verify that the protective effect of PLX5622 on retinal ganglion cells (RGCs) is not achieved by eliminating microglia, Iba1 immunofluorescence staining was performed on isolated retinal samples to evaluate the effect of PLX5622 on the number of microglia: Six UPOAO model mice were randomly selected, with three mice in each group. The healthy group was gavaged with PLX5622 at a dose of 40 mg PLX5622 per kg body weight per day for 7 days, and the healthy group was gavaged with an equal volume of control solvent for 7 days. After 7 days of gavage, the mice were euthanized, the eyeballs were removed, the cornea and iris were cut off with scissors, fixed in 4% paraformaldehyde solution for 1 hour, the eyeballs were rinsed with PBS, the retina was immersed in 5% BSAT solution, and placed in a 4°C refrigerator for blocking overnight. Iba1 rabbit serum primary antibody was incubated in a 4°C refrigerator for 48 hours, rinsed in PBS buffer, and incubated with AlexaFluor594 donkey anti-rabbit fluorescent secondary antibody at 4°C overnight. After rinsing in PBS buffer, the retina was flattened with the optic nerve layer facing up, sealed with anti-fluorescence quenching sealing solution, and photographed using a confocal microscope to count microglia. Figure 5 As shown, there was no statistical difference in the number of microglia in the retinal optic nerve layer of the PLX5622-treated healthy group of mice compared with the healthy group, confirming that PLX5622 did not eliminate the microglia in the retinal tissue when the dosage was 40 mg / kg / d, and also indicating that PLX5622 did not protect RAO nerves by eliminating microglia.

[0057] (4.2) In the in vitro experiment, the present invention used PLX5622 to intervene in the microglial cell line BV2 to further verify that PLX5622 did not work by reducing and eliminating microglia: Using high-glucose DMEM complete medium containing 10% fetal bovine serum and 1% double antibodies (penicillin and streptomycin), the PLX5622 working solution was diluted to a series of concentration gradients of 0.1, 0.25, 0.5, 1, and 5 μM. As a control of the experiment, the corresponding volume of the control solvent was diluted to the same dilution ratio as the 5 μM PLX5622 working solution as a negative control.

[0058] BV2 cells were cultured in the above culture medium for 24 hours, and the cell activity of each group was analyzed using the CCK8 cell activity detection kit to evaluate the effect of PLX5622 on the proliferation and activity of BV2 cells. Figure 6 As shown, the cell viability of BV2 cells was not affected in the culture medium with different PLX5622 concentrations, confirming that low-dose PLX5622 had no effect on the cell activity of the microglial cell line BV2 in vitro, and also confirming that PLX5622's protection of the mouse retina was not achieved by eliminating BV2 cells.

[0059] In the description of this specification, the description with reference to the terms "one embodiment / method", "some embodiments / methods", "example", "specific example" or "some examples" etc. means that the specific features, structures, materials or characteristics described in conjunction with the embodiment / method or example are included in at least one embodiment / method or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment / method or example. Moreover, the specific features, structures, materials or characteristics described may be combined in a suitable manner in any one or more embodiments / methods or examples. In addition, those skilled in the art may combine and combine the different embodiments / methods or examples described in this specification and the features of the different embodiments / methods or examples, unless they are contradictory.

[0060] It should be noted that, in the present invention, relational terms such as "first" and "second" are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Moreover, the terms "include", "comprise" or any other variants thereof are intended to cover non-exclusive inclusion, so that the process, method, article or equipment including a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or equipment. In the absence of further restrictions, the elements defined by the sentence "including one..." do not exclude the existence of other identical elements in the process, method, article or equipment including the elements. In the present invention, the meaning of "multiple" is at least two, such as two, three, etc., unless otherwise clearly and specifically specified.

[0061] The foregoing is merely a specific embodiment of the present invention, which enables those skilled in the art to understand or implement the present invention. Various modifications to these embodiments will be apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to the embodiments shown herein, but will conform to the widest scope consistent with the principles and novel features applied herein.

Claims

1. Use of PLX5622 or a pharmaceutically acceptable salt thereof in the preparation of a drug for preventing and treating retinal ischemia-reperfusion injury caused by retinal artery occlusion, resulting in retinal structural damage and / or functional damage.

2. The use according to claim 1, characterized in that: The damage to the retinal structure is caused by damage or death of retinal ganglion cells.

3. The use according to claim 1, characterized in that: The medicine is a medicine for treating retinal artery occlusion or a medicine for preventing complications after treatment of retinal artery occlusion.

4. The use according to claim 3, characterized in that: The therapeutic drug is an arterial thrombolytic drug, and PLX5622 or a pharmaceutically acceptable salt thereof is used as an auxiliary component in the arterial thrombolytic drug, and the auxiliary component is used to prevent retinal structural damage and / or functional damage.

5. The use according to claim 4, characterized in that: The arterial thrombolytic drug contains a fibrinolytic component and / or a vasodilator component as main components.

6. The use according to claim 3, characterized in that: The treatment of retinal artery occlusion includes one or more of anterior chamber puncture, acetazolamide treatment, eyeball massage, arterial thrombolytic drug thrombolysis, and interventional surgical thrombolysis.

7. The use according to claim 3, characterized in that: The complications are damage to the retinal structure and / or damage to its function.

8. The use according to claim 1, characterized in that: The PLX5622 is in the form of a pharmaceutically acceptable acid addition salt.

9. The use according to claim 1, characterized in that: The dosage form of the drug is one or more of an oral preparation, a microcapsule preparation, an injection, a suppository, an eye drop, an eye wash, an eye cream, and an eye gel.

10. The use according to claim 1, characterized in that: The symptom of structural damage is at least one of white lesions in the fundus and retinal edema; the symptom of functional damage is at least one of a sharp decrease in vision and visual field loss.

Images