Use of csf1r receptor and inhibitors thereof

Abstract

The present application relates to the field of biological medicine, and particularly relates to a new use of CSF1R and an inhibitor thereof, and discloses application of CSF1R as a target point in screening GABA A receptor, NMDA receptor, beta 2A receptor anesthetics intervention drugs. The present application also discloses application of a CSF1R inhibitor, PLX5622, in preparing GABA A , NMDA, beta 2A receptor anesthetics intervention drugs; and the present application can significantly shorten the duration of GABA A , NMDA, beta 2A receptor anesthetics induced loss of consciousness.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of biological medicine, in particular to the new use of CSF1R and its inhibitors. BACKGROUND

[0002] General anesthesia can induce reversible loss of consciousness and pain sensation, and is widely used in modern surgical operation and related medical examination. There are various general anesthetics, which are respectively directed to different neuronal receptors or molecular targets, including opioid receptors, γ-aminobutyric acid type A receptor (GABA A receptor complex: GABA receptor, benzodiazepine receptor and GABA receptor coupled chloride channel), N-methyl-D-aspartate (NMDA) receptor, β2 adrenergic receptor (β2A) and the like, to play the role of general anesthesia. In order to implement appropriate depth of anesthesia, several general anesthetics are often used in combination in clinic, which produce ideal anesthetic effect through comprehensive action with multiple neuronal receptors or molecular targets. Although modern anesthesia technology basically guarantees the safety and effectiveness during anesthesia, there are still a small number of patients who have a series of complications such as permanent damage to the central nervous system and postoperative delirium due to slow anesthetic recovery, which seriously affects the prognosis of patients.

[0003] The existing anesthetic acting on benzodiazepine receptor is flumazenil, and the opioid drug acting on opioid receptor is naloxone. The most commonly used general anesthetic in clinic is mainly directed to GABA A receptor, NMDA receptor and β2A receptor, and slow recovery of general anesthesia induced by such target anesthetics can cause serious central nervous system damage. However, there is currently a lack of intervention drugs for general anesthesia induced by such receptors, so it is of great clinical significance to find intervention drugs for GABA A receptor, NMDA receptor and β2A receptor anesthetics, which can help patients in anesthetic state induced by GABA A receptor, NMDA receptor or β2A receptor anesthetics to recover normal physiological activities.

[0004] CSF1R is encoded by the c-fms proto-oncogene, which is one of the growth factor receptors of intrinsic tyrosine-specific protein kinase activity, belongs to the platelet-derived growth factor (PDGF) family of factors, and is similar to other family members, with a highly glycosylated extracellular region composed of five immunoglobulin domains (D1-D5, 498 amino acids), a transmembrane domain (21 amino acids), an intracellular near-membrane domain composed of 36 amino acids, and an intracellular tyrosine kinase domain composed of 398 amino acids (inserted with a kinase of 73 amino acids). CSF1R has two natural ligands, namely interleukin-34 and CSF1 cytokine; the binding of the ligand can induce the dimerization of the homotypic receptor, autophosphorylation and the phosphorylation of downstream signals.

[0005] CSF-1R is expressed at a low level in hematopoietic stem cells, at a higher level in monocytes and tissue macrophages, and at a high level in osteoclasts, myeloid dendritic cells, microglia cells, etc., and controls the development, proliferation, differentiation, survival, etc. of these cells. In addition, it is also expressed in oocytes and pre-implantation embryos, decidual cells and trophoblast cells, neural progenitor cells and other neuronal cells, renal proximal tubular epithelial cells and colon epithelial cells. The wide expression pattern of CSF-1R is consistent with its pleiotropic role in embryonic development, adulthood, innate immunity, inflammation, tissue repair and tumor microenvironment.

[0006] Currently, CSF1R inhibitors have been used for the treatment of various diseases, including tenosynovial giant cell tumor, pigmented villonodular synovitis (clinical phase 3), solid tumors (clinical phase 1), graft-versus-host disease (clinical phase 3), idiopathic pulmonary fibrosis (clinical phase 1), etc.; in the central nervous system, mainly for cerebral hemorrhage (clinical phase 1), Alzheimer's disease (clinical phase 1), cognitive impairment (clinical phase 2), amyotrophic lateral sclerosis (clinical phase 1), brain tumors (glioma, meningioma, pituitary tumor), etc. Recently, the CSF-1R inhibitor Pimicotinib (ABSK021) of Xiyu Medicine has been granted a breakthrough therapy designation by the US Food and Drug Administration for the treatment of inoperable tenosynovial giant cell tumor.

[0007] The invention of CN114107196A, "Method for differentiating natural killer cells from pluripotent stem cells based on single-cell sequencing rational design and application of CSF1R inhibitor", discloses the application of CSF1R inhibitors in promoting the differentiation of pluripotent stem cells in vitro to prepare natural killer cells. The CSF1R inhibitors include but are not limited to BLZ945 and / or PLX3397, for example, Linifanib (ABT-869), OSI-930, GW2580, PLX5622, etc.

[0008] The application of CN110404078B, a combined treatment method for alleviating microglia / macrophage-mediated inflammatory response after spinal cord injury, provides the use of biodegradable hydrogel materials for transplantation and CSF1R inhibitors in the preparation of products for alleviating microglia / macrophage-mediated inflammatory response after spinal cord injury. The CSF1R inhibitor is one or more selected from PLX3397, PLX5622, and GW2580.

[0009] Currently, there is no related technology report on the correlation between microglial CSF1R and general anesthesia and anesthesia recovery. SUMMARY

[0010] The technical problem to be solved by the present application is to provide a new use of CSF1R and its inhibitors.

[0011] To solve the above technical problems, the present application provides the use of CSF1R as a target in screening GABA A receptor, NMDA receptor, and beta2A receptor class anesthetics.

[0012] The present application also provides the use of CSF1R inhibitors in the preparation of GABA A , NMDA, and beta2A receptor class anesthetics.

[0013] As an improvement of the application of the present application, the CSF1R inhibitor is PLX5622.

[0014] As a further improvement of the application of the present application, the GABA A , NMDA, and beta2A receptor class anesthetics induced general anesthesia effect is intervened.

[0015] As a further improvement of the application of the present application, the duration of loss of consciousness induced by GABA A , NMDA, and beta2A receptor class anesthetics is shortened (significantly shortened).

[0016] As a further improvement of the application of the present application, the GABA A receptor class anesthetics is pentobarbital sodium, the NMDA receptor class anesthetics is ketamine, and the beta2A receptor class anesthetics is dexmedetomidine.

[0017] In summary, the present application provides the use of CSF1R inhibitor PLX5622 in the intervention of GABA A , NMDA, and beta2A receptor class anesthetics induced general anesthesia effect.

[0018] Through extensive and in-depth research, the inventors have discovered for the first time that treatment with the CSF1R inhibitor PLX5622 plays a crucial regulatory role in maintaining general anesthesia. Treatment of mice with the clinically tested CSF1R inhibitor PLX5622 for one week significantly shortened GABA levels. A The duration of general anesthesia induced by different types of receptor anesthetics such as NMDA and β2A was studied. This study lays the foundation for future development of GABA. A The development of arousal-promoting drugs based on receptor, NMDA receptor, and β2A receptor anesthetics laid an important foundation. Approximately one week before the injection of the anesthetic, the CSF1R inhibitor PLX5622 is administered orally, followed by intraperitoneal administration of appropriate general anesthetics such as sodium pentobarbital, ketamine, or dexmedetomidine, as needed. Attached Figure Description

[0019] The specific embodiments of the present invention will be further described in detail below with reference to the accompanying drawings.

[0020] Figure 1 The CSF1R receptor inhibitor PLX5622 can significantly shorten the time to pentobarbital sodium (GABA). A General anesthesia induced by receptor anesthetics;

[0021] Figure 1 middle:

[0022] A is a flowchart of the experiment, including the feeding duration of the two groups of mice, AIN-76 and PLX5622, the time points of anesthesia induction and recording;

[0023] Treatment with CSF1R receptor inhibitor PLX5622 (B) does not affect the time to onset of general anesthesia induced by sodium pentobarbital.

[0024] Treatment with C, the CSF1R receptor inhibitor PLX5622, significantly shortened the duration of general anesthesia induced by sodium pentobarbital.

[0025] Figure 2 The CSF1R receptor inhibitor PLX5622 can significantly shorten the duration of ketamine-induced general anesthesia (an NMDA receptor anesthetic).

[0026] Figure 2 middle:

[0027] A is a flowchart of the experiment, including the feeding duration of the two groups of mice, AIN-76 and PLX5622, the time points of anesthesia induction and recording;

[0028] Treatment with the CSF1R receptor inhibitor PLX5622 (B) does not affect the time of entry into ketamine-induced general anesthesia.

[0029] C. Treatment with the CSF1R receptor inhibitor PLX5622 significantly shortened the duration of ketamine-induced general anesthesia.

[0030] Figure 3 B. Treatment with the CSF1R receptor inhibitor PLX5622 did not affect the onset of dexmedetomidine (a β2A receptor anesthetic) induced general anesthesia.

[0031] Figure 3 C. Treatment with the CSF1R receptor inhibitor PLX5622 significantly shortened the duration of dexmedetomidine-induced general anesthesia.

[0032] A. Flow chart of the experiment, including the duration of feeding of AIN-76 and PLX5622 feed, induction of anesthesia, and time points recorded for each group of mice.

[0033] B. Treatment with the CSF1R receptor inhibitor PLX5622 did not affect the onset of dexmedetomidine-induced general anesthesia.

[0034] C. Treatment with the CSF1R receptor inhibitor PLX5622 significantly shortened the duration of dexmedetomidine-induced general anesthesia. DETAILED DESCRIPTION

[0035] The application will be further described below in conjunction with specific embodiments, but the scope of protection of the application is not limited to this:

[0036] Example 1: Effect of CSF1R inhibitor PLX5622 treatment on GABA A receptor anesthetics

[0037] (1) Experimental animals

[0038] Eight-week-old SPF C57bl / 6j male mice were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. and the animals were raised in a SPF environment.

[0039] (2) Treatment with the CSF1R inhibitor PLX5622

[0040] The C57Bl / 6j male mice were divided into two groups (6 mice in each group), one group was fed with AIN-76A feed containing PLX5622 (1,200 mg / kg), which was called the PLX5622 group; the other group was fed with AIN-76A feed as a control group, and the animals in both groups were fed for one week before the experiment. During feeding, the feed was freely taken.

[0041] (3) Anesthetic injection and anesthesia duration statistics

[0042] On the day of the experiment, the body weight of each animal was weighed, and then GABA AThe animals were placed under a video recorder after receiving the receptor anesthetic sodium pentobarbital (100 mg / kg of sodium pentobarbital, dissolved in 0.2 ml of normal saline); then the specific time at which each mouse lost the righting reflex was checked, and the specific time at which each mouse recovered the righting reflex was recorded by video, and the duration of each mouse's loss of consciousness induced by general anesthesia was calculated.

[0043] (4) Statistical method

[0044] The data are expressed as mean ± standard error (SEM), and statistical significance analysis was performed using Graphpad Prism. The comparison of two groups of data used unpaired student's t test. In all statistical results, n.s. indicates no significant difference, p < 0.05 is considered to be a significant difference, * indicates p < 0.05, ** indicates p < 0.01, and *** indicates p < 0.001.

[0045] (5) Experimental results

[0046] The same age control group and PLX5622 group mice without other treatments were selected, the body weight of each mouse was weighed, and then GABA A The animals were placed under a video recorder after receiving the receptor anesthetic sodium pentobarbital (100 mg / kg), and the anesthesia process was recorded by video Figure 1 A) During the induction of anesthesia, the time point at which each mouse lost the righting reflex (loss of righting reflex, LORR, i.e., entered general anesthesia) was checked manually; and the time point at which the righting reflex was recovered (recovery of righting reflex, RORR, i.e., woke up from general anesthesia) was determined by video, and the duration of LORR and RORR was calculated and analyzed, respectively.

[0047] It was found that the use of the CSF1R inhibitor PLX5622 could significantly shorten the duration of RORR induced by sodium pentobarbital, and there was no significant statistical difference in the duration of LORR entering general anesthesia Figure 1 B-C).

[0048] Example 2: Effect of CSF1R inhibitor PLX5622 treatment on NMDA receptor anesthetics

[0049] (1) Experimental animals

[0050] Eight-week-old SPF C57bl / 6j male mice were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd., and the animals were raised in a SPF environment.

[0051] (2) Treatment of CSF1R inhibitor PLX5622

[0052] C57B1 / 6j male mice were divided into two groups (6 mice in each group), one group was fed with AIN-76A feed containing PLX5622 (1,200 mg / kg), called PLX5622 group; the other group was fed with AIN-76A feed as control group, and the animals in both groups were fed with food for one week before the experiment. During feeding, the feed was free to take.

[0053] (3) Anesthesia injection and anesthesia duration statistics

[0054] On the day of the experiment, the body weight of each animal was weighed, and then ketamine, an NMDA receptor anesthetic, was used intraperitoneally according to the body weight (100 mg of ketamine / kg of body weight, dissolved in 0.2 ml of normal saline), and the animals were placed under a video recorder; then the specific time for each mouse to lose the righting reflex was checked, and the specific time for each mouse to recover the righting reflex was recorded by video, and the duration of each mouse entering the induction of general anesthesia was calculated.

[0055] (4) Statistical method

[0056] The data are expressed as mean ± standard error (SEM), and the statistical significance analysis is performed using Graphpad Prism. The comparison of two groups of data is performed using unpaired student's t test. In all statistical results, n.s. indicates no significant difference, p<0.05 is considered to be a significant difference, * indicates p<0.05, ** indicates p<0.01, and *** indicates p<0.001.

[0057] (5) Experimental results

[0058] The same age control group and PLX5622 group mice without other treatment were selected, the body weight of each mouse was weighed, then ketamine, an NMDA receptor anesthetic, was used intraperitoneally according to the body weight of each mouse (100 mg / kg), and the anesthetic process was recorded under a video recorder Figure 2 A). During the induction of anesthesia, the time point for each mouse to lose the righting reflex (LORR, i.e. enter general anesthesia) was checked manually; and the time point for recovery of righting reflex (RORR, i.e. awakening from general anesthesia) was determined by video, and the duration of LORR and RORR was calculated and analyzed respectively.

[0059] It was found that the use of CSF1R inhibitor PLX5622 could significantly shorten the duration of RORR of ketamine-induced general anesthesia, while there was no significant statistical difference in LORR of entering general anesthesia Figure 2 B-C).

[0060] Example 3: Effect of CSF1R inhibitor PLX5622 treatment on beta2A receptor anesthetics

[0061] (1) Experimental animals

[0062] Eight-week-old SPF C57bl / 6j male mice were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. and the animals were raised in a SPF environment.

[0063] (2) Treatment of CSF1R inhibitor PLX5622

[0064] The C57Bl / 6j male mice were divided into two groups (3 in each group), one group was fed with AIN-76A feed containing PLX5622 (1,200 mg / kg), called the PLX5622 group; the other group was fed with AIN-76A feed as a control group, and the animals in both groups were fed with food for one week before the experiment. During feeding, the feed was freely taken.

[0065] (3) Anesthetic injection and anesthesia duration statistics

[0066] On the day of the experiment, the body weight of each animal was weighed, and then the beta2A receptor anesthetic dexmedetomidine (0.2 mg dexmedetomidine / kg body weight, dissolved in 0.2 ml of normal saline) was used intraperitoneally according to the body weight of each mouse. The animals were placed under a video recorder; then the specific time for each mouse to lose the righting reflex was checked, and the specific time for each mouse to recover the righting reflex was recorded by video, and the duration of loss of consciousness for each mouse to enter general anesthesia induction was calculated.

[0067] (4) Statistical method

[0068] The data is expressed as mean ± standard error (SEM), and the statistical significance analysis is performed using Graphpad Prism. The comparison of two groups of data is performed using unpaired student's t test. In all statistical results, n.s. indicates no significant difference, p<0.05 is considered to be a significant difference, * indicates p<0.05, ** indicates p<0.01, and *** indicates p<0.001.

[0069] (5) Experimental results

[0070] The same age control group and PLX5622 group mice without other treatment were selected, the body weight of each mouse was weighed, and then the beta2A receptor anesthetic dexmedetomidine (0.2 mg / kg) was used intraperitoneally according to the body weight of each mouse, and the anesthesia process was recorded under a video recorder Figure 3A) During the induction phase of anesthesia, the time point of loss of righting reflex (LORR, i.e. entering general anesthesia) was manually checked for each mouse; and the time point of recovery of righting reflex (RORR, i.e. awakening from general anesthesia) was determined by video, the LORR and RORR durations were calculated and analyzed, respectively.

[0071] It was found that the use of CSF1R inhibitor PLX5622 could significantly shorten the RORR duration induced by the beta2A receptor anesthetic dexmedetomidine, while there was no significant statistical difference in the LORR of entering general anesthesia Figure 3 B-C).

[0072] Finally, it should also be noted that the above only lists several specific embodiments of the present application. Obviously, the present application is not limited to the above embodiments, but can also have many variations. All variations that can be directly derived or inferred from the disclosure of the present application by those of ordinary skill in the art should be considered as falling within the scope of the present application.

Claims

1. Use of a CSF1R inhibitor in the manufacture of a medicament for the intervention of GABA A , NMDA or beta2A receptor class anesthetics, characterized in that: The CSF1R inhibitor is PLX5622, the intervening drug is for shortening the length of GABA A , NMDA or beta2A receptor class anesthetic induced loss of consciousness; GABA A The receptor class of anesthetics was pentobarbital sodium, the NMDA receptor class of anesthetics was ketamine, and the β2A receptor class of anesthetics was dexmedetomidine.

2. Use according to claim 1, characterized in that: Intervention of GABA A , NMDA or β2A receptor class anesthetics to induce general anesthesia.

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