Application of LP17 in preparation of intervention medicine for preventing / treating autism

Abstract

The invention relates to an application of LP17 in preparation of an intervention medicine for preventing / treating autism, the LP17 can inhibit TREM1 expression and can reduce central nervous system inflammation, and tests prove that the LP17 can inhibit TREM1 accumulation and significantly reduce neural immune inflammation of an autism model BTBR mouse when acting on the LP17 in the early stage of mouse after birth, and the LP17 can be used for preventing / treating the autism of the mouse after the birth of the mouse after the birth of the mouse after the birth of the mouse after the birth of the mouse after the birth of the mouse. According to the present invention, the mouse model has characteristics of low cost, low cost, neuroprotection and adult mouse behavioral correction ability, improves the defects of the three-room social experiment of the adult mouse, reduces the inscription behavior of the adult mouse, reverses the influence of the autism related behavior of the adult mouse, and provides the clue for the development of the autism.

Description

Technical Field

[0001] The present invention belongs to the field of biomedical technology, and specifically relates to the application of LP17 in the preparation of intervention drugs for preventing / treating autism. Background Art

[0002] Autism is classified as a developmental disorder caused by a disorder of the nervous system, with symptoms including social dysfunction and limited, repetitive behaviors or interests. Currently, there is a growing demand for professional treatment, rehabilitation and educational services for autistic patients and their families. Patients and their families hope to obtain more effective treatments, more professional rehabilitation guidance and more comprehensive educational support to improve the quality of life and social adaptability of patients.

[0003] BTBR mice are currently recognized as an animal model of idiopathic autism. Their core symptoms are most similar to the clinical symptoms of autistic patients and can be stably inherited to offspring. BTBR mice showed obvious social dysfunctions such as lack of social motivation and social ability in the three-chamber social experiment. In the direct social interaction experiment, BTBR mice had significantly reduced exploratory social behaviors such as sniffing and following. In the USV test (ultrasonic vocalization test), when BTBR mice left the cage of the mother mouse, the BTBR mice emitted abnormally high-frequency and high-amplitude ultrasonic waves. Researchers believe that this behavior is similar to the loud crying of autistic children when they are forced to separate from their mothers. In contrast, adult BTBR mice emit a reduced number of ultrasonic waves when faced with the smell of unfamiliar mice, which is similar to the language communication disorders of human autistic patients. In addition, studies have shown that BTBR mice have increased repetitive behaviors, such as living alone, digging, grooming, and burying beads. The anatomical characteristics of BTBR mice are also consistent with those of autistic patients. For example, in BTBR mice, organic lesions in the central nervous system are mainly concentrated in the cortex, corpus callosum, and hippocampal hypoplasia, features that are similar to the clinical manifestations of autism patients.

[0004] Studies have shown that the occurrence of autism is related to chronic inflammation of the central nervous system, and the exact pathogenesis has been determined to have reliable diagnostic or prognostic value for autism. Multiple lines of evidence have shown that infectious or inflammatory immune activation during pregnancy leads to a higher risk of autism-related neurodevelopmental symptoms in offspring. It was found that the offspring of maternal immune activation (MIA) model mice exhibited autistic-like behaviors and cortical damage caused by MIA, which reduced the overproduction of CTIP2+ (cortical thymocyte transformation protein 2-positive) cortical neurons in the late postnatal period. This suggests that the phenotype caused by persistent neuroinflammation after birth is a key risk factor for inducing autism-related behaviors.

[0005] Triggering receptor expressed on myeloid cells-1 (TREM1) is a receptor belonging to the immunoglobulin superfamily that triggers the synthesis and release of proinflammatory cytokines and plays an important role in acute and chronic inflammation. The important role of regulating neuroinflammation in autism by inhibiting the accumulation of TREM1 is a previously unappreciated mechanism.

[0006] From the current research status, the exact cause of autism is still unclear, and there is no specific drug. Only certain antipsychotic drugs can help relieve the symptoms of autism. At present, the treatment of autism is mainly behavioral therapy. The core symptoms of some autistic patients will continue into adulthood and require lifelong support.

[0007] In summary, the discovery of a drug that can directly target the core symptoms of autism and effectively improve them will be of great significance for the treatment of autism and for improving adult patients from being troubled by autism symptoms. Summary of the invention

[0008] In view of the deficiencies in the prior art, the present invention proposes the use of LP17 in the preparation of intervention drugs for preventing / treating autism.

[0009] The technical solution of the present invention is: Application of LP17 in the preparation of intervention drugs for the prevention / treatment of autism.

[0010] Furthermore, the LP17 can inhibit TREM1 expression.

[0011] Furthermore, the LP17 can reduce central nervous system inflammation.

[0012] Furthermore, the drug acts on autistic patients in the early postnatal period.

[0013] Furthermore, the drug is administered by intracranial injection.

[0014] Furthermore, the effective dosage of the drug is 20 mg / kg.

[0015] Furthermore, the drug contains pharmaceutically acceptable excipients.

[0016] The auxiliary materials include, but are not limited to, diluents, buffers, suspensions, emulsions, granules, encapsulation agents, excipients, fillers, adhesives, sprays, transdermal absorbents, wetting agents, disintegrants, absorption enhancers, surfactants, colorants, flavoring agents or adsorption carriers.

[0017] Compared with the prior art, the present invention has at least the following advantages: The present invention relates to the use of LP17 in the preparation of an intervention drug for preventing / treating autism. The LP17 can inhibit TREM1 expression and reduce central nervous system inflammation. Experiments have shown that LP17 can inhibit TREM1 accumulation in the early postnatal period of mice, significantly reduce neuroimmune inflammation in autism model BTBR mice, and has neuroprotection and behavioral correction capabilities in adult mice. It significantly reduces neuronal apoptosis in the cortex of BTBR mice higher than that of wild-type mice, improves the defects of a three-chamber social experiment in adult mice, reduces stereotyped behaviors in adult mice, and reverses the effects of autism-related behaviors in adult mice, providing clues for the development of autism. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] In order to more clearly illustrate the specific embodiments of the present invention, the drawings required for use in the specific embodiments or the description of the prior art will be briefly introduced below.

[0019] Figure 1 This is the enrichment of KEGG pathway in test example 1 of the present invention; Figure 2 This is the test example 1 of the present invention, ELISA and RT-qPCR to detect the expression of inflammatory factors; Figure 3 This is the activation degree of microglia in the cortex of BTBR mice in Test Example 2 of the present invention; Figure 4 This is the neuronal apoptosis in the cortex of BTBR mice in Test Example 2 of the present invention; Figure 5 This is the test example 3 of the present invention. The expression level of TREM1 protein in the cerebral cortex of P7 BTBR-Vehicle mice was significantly upregulated; Figure 6 The results of comparative analysis of homology and functional similarity of human and mouse TREM1 genes and proteins in test example 4 of the present invention are as follows; Figure 7 For the fourth test example of the present invention, a comparative analysis of the functions and interacting proteins of human and mouse TREM1 proteins was performed on the bioinformatics; Figure 8 For the test example 4 of the present invention, the core function enrichment of human and mouse TREM1 proteins was analyzed by bioinformatics; Fig. 9 For the test example of the present invention, five pairs of BTBR-Vehicle, BTBR-LP17, and C57BL / 6-Vehicle group mice at 7 days after birth were tested for changes in the activation state of mouse cortical microglia and the level of TREM1 protein expressed by microglia before and after medication; Fig.10 The total amount of TREM1 protein expression in brain tissue of the three groups of mice in Test Example 5 of the present invention was detected by ELISA before and after medication; Fig.11 The expression of pro-inflammatory factors in the cerebral cortex tissue of the four groups of mice in Test Example 6 of the present invention before and after medication; Fig.12 This is an analysis of the changes in the number of apoptotic neurons in the cerebral cortex of mice before and after medication in Test Example 6 of the present invention; Fig.13 This is the three-chamber social experiment test analysis of the four groups of mice before and after medication in Test Example 7 of the present invention; Fig.14 This is a test analysis of the glass bead burying and self-grooming behaviors of the four groups of mice in Test Example 7 of the present invention before and after medication. DETAILED DESCRIPTION

[0020] The present invention is further described in detail below. It is necessary to point out that the following specific implementation methods are only used to further illustrate the present invention and cannot be understood as limiting the scope of protection of the present invention. Technicians in this field can make some non-essential improvements and adjustments to the present invention based on the above application content.

[0021] The present invention provides a general and / or specific description of the materials and test methods used in the experiment. The test methods or test methods involved are all conventional methods unless otherwise specified; the reagents or instruments used are all conventional products available on the market, prepared or used in a conventional manner, unless the manufacturer is specified.

[0022] Sources: BTBR mice (autism mice), male and female BTBR T+Itpr3tf / J (BTBR) strain breeding colonies were purchased from Jackson Laboratory (Bar Harbor, ME, USA) and used in the experiments of this patent after breeding; C57BL / 6 mice (wild-type mice) were purchased from the Army Medical University Laboratory; Housing environment: 5-6 animals per cage were grouped according to a 12-h light-dark cycle and housed in a standard animal facility with free access to water and food.

[0023] LP17 (an inhibitory peptide targeting TREM1, with an amino acid sequence of LQVTDSGLYRCVIYHPP (SEQ ID NO. 3)) was synthesized by Chongqing Biaoyue Biotechnology Co., Ltd.

[0024] Example Pretreatment of BTBR mice and C57BL / 6 mice BTBR mice were randomly divided into two groups, 10 in each group, and the birth date was set as postnatal day 0 (P0). To avoid litter effect, animals in each treatment group were randomly selected from multiple litters, and one pup was selected from each litter. As a control group, C57BL / 6 mice were treated in the same way.

[0025] Group ①: BTBR-Vehicle (solvent control) group mice were treated with intracerebroventricular injection of 0.9% NaCl:DMSO=2:3 at P5 (postnatal day 5) and P6 (postnatal day 6).

[0026] Group ②: BTBR-LP17 group mice were treated with intracerebroventricular injection of LP17 at a concentration of 20 mg / kg at P5 and P6.

[0027] Group ③: C57BL / 6-Vehicle group mice were injected with 0.9% NaCl:DMSO=2:3 into the lateral ventricle at P5 and P6.

[0028] Group ④: C57BL / 6-LP17 group mice were treated with intracerebroventricular injection of LP17 at a concentration of 20 mg / kg at P5 and P6.

[0029] Various tests were performed on the decimals grouped above, and the test results are as follows.

[0030] Test Example 1: Exacerbated cortical inflammation in P7-BTBR mice (7-day-old BTBR mice) ① In this test case, cortical tissues of P7 BTBR-Vehicle and P7 C57BL / 6-Vehicle mice were collected for transcriptome sequencing. Bioinformatics analysis was performed and functional annotations of DEGs (differentially expressed genes) were used based on the Kyoto Encyclopedia of Genes and Genomes (KEGG) database and the Gene Ontology (GO) category database. Changes in gene mRNA expression in cortical tissues of BTBR mice in the early postnatal period were detected at the transcriptome level. The test results are as follows: Figure 1 As shown in the figure, the pathway enrichment of KEGG pathways with a P value less than 0.05 is considered significant. The pathways are sorted by P value, and the top 20 pathways are displayed in a bar graph with -log10 (P value) on the horizontal axis. The bar graph is colored according to the proportion of up-regulated or down-regulated differentially expressed genes; the significantly enriched gene ontology entries are displayed in the form of dot graphs, and the points are colored continuously according to -log10 (P value), from light to dark, indicating that the P value is from low to high, and the size of the point varies according to the number of differentially expressed genes belonging to the entry; from Figure 1 As can be seen, KEGG showed that the immune system processes and inflammatory responses of P7 BTBR-Vehicle mice were significantly enriched compared with P7 C57BL / 6-Vehicle mice.

[0031] ② Through ELISA (enzyme-linked immunosorbent assay) and RT-qPCR detection (reverse transcription real-time fluorescence quantitative polymerase chain reaction), the results showed that compared with P7 C57BL / 6-Vehicle, the mRNA expression levels of IL-6 (‌interleukin-6) and TNF-α (tumor necrosis factor α) in the brain of P7 BTBR-Vehicle mice were significantly increased; ELISA experiments were performed, and the test results confirmed that the protein levels of proinflammatory factors IL-6, TNF-α, and CXCR2 (CXC chemokine receptor 2) in the cortex of P7 BTBR-Vehicle mice were significantly higher than those in the control group mice (C57BL / 6-Vehicle). At the same time, the ELISA results also confirmed that the expression of the inflammatory factor NF-κB (nuclear factor κB) protein level in the cortex of P7 BTBR-Vehicle mice was also significantly upregulated ( Figure 2 ), further confirming that the neuroimmune inflammatory microenvironment in the brain of BTBR mice changes during early postnatal life.

[0032] Test Example 2: The activation of microglia in the cortex of P7-BTBR-Vehicle mice increased significantly, and the apoptosis of neurons in the cortex increased significantly compared with the control group mice. ① In this test case, immunofluorescence staining was used to observe the activation of microglia, resident immune cells in the cortex of P7 BTBR-Vehicle mice; the results are as follows Figure 3 As shown, Figure 3 A. Figure 3 The results in B showed that compared with P7 C57BL / 6-Vehicle mice, the number of activated microglia in the cerebral cortex of P7 BTBR-Vehicle mice increased significantly. Figure 3 As can be seen in C, the cell morphology is mostly ameba-like, with rounded cell bodies and shortened branches.

[0033] ② Changes in the immune inflammatory microenvironment in the brain of early postnatal BTBR mice may affect neuronal apoptosis. Therefore, this test case used immunofluorescence staining to observe the cortical neurons of P7 BTBR-Vehicle mice; the experimental results are shown in Figure 2. Figure 4As shown in the figure, compared with P7 C57BL / 6-Vehicle mice, the number of NEUN+TUNEL+ double-positive cells in the cerebral cortex of P7 BTBR-Vehicle mice increased significantly (‌NEUN+TUNEL+ double labeling technology‌ is a method for simultaneously detecting neuronal apoptosis and neuronal existence, NEUN (Neuronal Nuclei) is a neuronal nuclear protein, and TUNEL (Terminal deoxynucleotidyl transferase dUTP nick end labeling) technology is used to detect cell apoptosis), indicating that the number of neuronal apoptotic cells in the brain of BTBR mice in the early postnatal period increased significantly.

[0034] Test Example 3: TREM1 expression significantly increased in the cortex of P7 BTBR-Vehicle mice ① In this test case, transcriptome sequencing was performed on the cerebral cortex of P7 BTBR-Vehicle mice, and bioinformatics analysis was performed on the differentially expressed genes in the cortex of P7 BTBR-Vehicle mice and P7 C57BL / 6-Vehicle mice. The results are as follows: Figure 5 As shown in A, it can be seen from the figure that upregulated TREM1 expression is of great significance in the differential gene expression network in the cortex of BTBR and control group mice.

[0035] ② This test case further tested the TREM1 protein expression level by ELISA. The test results are as follows: Figure 5 As shown in B, it was confirmed that the expression level of TREM1 protein in the cortex of P7 BTBR-Vehicle mice was significantly upregulated compared with that in P7 C57BL / 6-Vehicle mice.

[0036] Comparative analysis of homology and functional similarity of human and mouse TREM1 genes and proteins in test case 4 ① Obtain human and mouse TREM1 gene and protein sequences from NCBI and Uniprot databases.

[0037] Human TREM1 protein sequence: MRKTRLWGLLWMLFVSELRAATKLTEEKYELKEGQTLDVKCDYTLEKFASSQKAWQIIRDGEMPKTLACTERPSKNSHPVQVGRIILEDYHDHGLLRVRMVNLQVEDSGLYQC VIYQPPKEPHLMLFDRIRLVVTKGFSGTPGSNENSTQNVYKIPPTTTKALCPLYTSPRTVTQAPPKSTADVSTPDSEINLTNVTDIIRVPVFNIVILLAGGFLSKSLVFSVLFAVTLRSFVP (SEQ ID NO.1) Mouse TREM1 protein sequence: MRKAGLWGLLCVFFVSEVKAAIVLEEERYDLVEGQTLTVKCPFNIMKYANSQKAWQRLPDGKEPLTLVVTQRPFTRPSEVHMGKFTLKHDPSEAMLQVQMTDLQVTDSGLY RCVIYHPPNDPVVLFHPVRLVVTKGSSDVFTPVIIPITRLTERPILITTKYSPSDTTTTRSLPKPTAVVSSPGLGVTIINGTDADSVSTSSVTISVICGLLSKSLVFIILFIVTKRTFG (SEQ ID NO.2) ② Through prediction and BLAST sequence comparison (Basic Local Alignment Search Tool, a search tool based on local sequence alignment algorithm), Figure 6 As can be seen in A and 6B, the human TREM1 gene is located on chromosome 6p21.1, and its gene structure contains 5 exons, encoding a single transmembrane protein. The mouse TREM1 gene is located on chromosome 17 (corresponding to the homologous region of human chromosome 6p21.1) and is also composed of 5 exons. The gene structures of the two are highly conserved, and the number of exons and the protein transmembrane topology are consistent. Through BLAST comparison, the amino acid sequence homology of human and mouse TREM1 is about 60%; in terms of key regions, the TREM1 proteins of humans and mice are highly conserved in key ligand binding sites, such as the hydrophobic core region of the β-pleated sheet and other core functional regions (Uniprot: mouse TREM1: Q9JK2, human TREM1: Q9NP99); and the human and mouse protein structures are predicted by SWISS-MODEL, Figure 6 C is the predicted structure of mouse TREM1 protein. Figure 6 D is the prediction of the human TREM1 protein structure.

[0038] ③ STRING was used to conduct a comparative analysis of the functions and interacting proteins of human and mouse TREM1 proteins. The results are as follows: Figure 7 As shown, Figure 7 A is the prediction of mouse TREM1 interacting proteins, Figure 7 B is the prediction of human TREM1 interacting proteins. As can be seen from the figure, TYPOBP, TREM2, TREML2, TREML1, CLEC5A, PGLYRP1, and HMGB1 are present in both human TREM1 and mouse TREM1 interacting proteins, indicating that the core interacting molecules of human and mouse TREM1 are highly similar.

[0039] ④ Protein function enrichment analysis by Uniprot bioinformatics, Figure 8 A is the core function of human TREM1 protein. Figure 8 B is the core function of mouse TREM1 protein. As can be seen from the figure, the core functions of human and mouse TREM1 proteins are highly conserved, and the two are almost identical in terms of regulation of apoptotic cell clearance, regulation of interleukin-10 production, neuron apoptosis, and glial cell immune response, indicating that the core functions of human and mouse TREM1 are highly conserved.

[0040] Test Example 5: The expression of TREM1 in microglia in the cortex of P7 BTBR-Vehicle mice was significantly increased; while the TREM1-specific antagonist peptide LP17 significantly reduced the expression of TREM1 in the cortex of BTBR mice, and significantly reduced the activation level of microglia in the cortex of P7-BTBR-Vehicle mice.

[0041] ① The cortex of BTBR-Vehicle, BTBR-LP17, and C57BL / 6-Vehicle mice at 7 days after birth (P7) was stained using immunofluorescence technology. The stained cells were observed and photographed using a Zeiss (Oberkochen, Germany) Axivert microscope connected to the Zeiss AvioVision 3.0 system to detect the activation state of mouse cortical microglia (detection of IBA1) and changes in TREM1 expression levels.

[0042] The fluorescence detection results are as follows Fig. 9 As shown, from Fig. 9 As shown in A, the number of abnormally activated cortical microglia in the C57BL / 6-Vehicle group of mice was small, and the expression of TREM1 molecules was low; while the number of abnormally activated cortical microglia in the BTBR-Vehicle group of mice was significantly higher than that in the C57BL / 6-Vehicle group, and TREM1 protein was specifically and highly expressed in the microglia in the cortex of P7 BTBR-Vehicle mice. ELISA experiments also confirmed that the TREM1 protein in the microglia in the cortex of P7-BTBR mice was significantly higher than that in the C57BL / 6-Vehicle group of mice. The results of the ELISA experiment are shown in Figure 1. Fig. 9 As shown in B.

[0043] ② At the same time, ELISA was used to detect the total amount of TREM1 protein in the cerebral cortex of P7 BTBR-Vehicle, P7 BTBR-LP17, and P7 C57BL / 6-Vehicle mice. Fig.10 It can be seen that after LP17 treatment, the total expression of TREM1 protein in the brain tissue of P7 BTBR-Vehicle mice was significantly decreased.

[0044] The above results confirmed that after LP17 treatment, the expression of TREM1 in the cortex of P7-BTBR mice was significantly decreased, and the level of neuroimmune inflammation in the cortex was significantly reduced. LP17 can effectively inhibit the TREM1 pathway in the cortex of P7-BTBR mice and reduce the level of neuroimmune inflammation.

[0045] Test case 6 Early postnatal use of the specific peptide antagonist LP17 can effectively reduce the expression of pro-inflammatory factors in the cortex of P7 BTBP mice, that is, reduce the level of cortical inflammation in BTBR mice; at the same time, it can effectively reduce neuronal apoptosis in the cortex of BTBR mice, which is significantly higher than that of wild-type mice. ① The cerebral cortex tissues of BTBR-Vehicle, BTBR-LP17, C57BL / 6-Vehicle, and C57BL / 6-LP17 mice on postnatal day 7 (P7) were collected, and the protein expression levels of pro-inflammatory factors in the cerebral cortex, including interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and monocyte chemoattractant protein 1 (MCP-1), were detected by ELISA.

[0046] Test results such as Fig.11As shown in the figure, it can be seen that compared with C57BL / 6-Vehicle, the expression of pro-inflammatory factors in the cortex of BTBR-Vehicle mice was abnormally increased, and the protein expression levels of TNF-α, IL-6 and chemokine MCP-1 in the cortical tissue of BTBR mice were significantly increased (p<0.05); compared with BTBR-Vehicle, BTBR-LP17 was intervened with TREM1-specific peptide antagonist LP17 in the early postnatal period, which could significantly reduce the protein expression levels of TNF-α, IL-6 and MCP-1.

[0047] ② Using immunofluorescence experiments, it was confirmed that the use of a specific peptide antagonist LP17 in the early postnatal period could significantly reduce the number of excessively apoptotic neurons in the cerebral cortex of P7-BTBR mice. Fig.12 shown.

[0048] Therefore, combined with the results of Test Example 5, it was shown that the use of the specific peptide antagonist LP17 in the early postnatal period can effectively reduce the expression of TREM1 protein and inhibit the excessive immune inflammation of the central nervous system of BTBR mice, while reducing the excessive apoptosis of cortical neurons in BTBR mice and effectively inhibiting the TREM1 pathway.

[0049] Test case 7 Early postnatal inhibition of TREM1 improves autism-like classic behaviors in adult BTBR mice This test case uses classic autism-like behavioral experiments such as the three-chamber social test, bead burying test, and grooming test to evaluate whether the autism-like behavior of BTBR mice in adulthood (8 weeks after birth) is improved after intervention with the TREM1-specific antagonist LP17 in early postnatal life.

[0050] ①Three-room social experiment The social approach test was conducted in a rectangular apparatus (40 cm * 60 cm * 22 cm) divided into three equal parts. Mice were allowed to enter the side chambers through retractable doorways in two partition walls. The test consisted of habituation and socialization. During habituation, the animals were placed in the middle chamber and allowed to freely explore all three chambers. For socialization, a stranger mouse aged 8-10 weeks, matched for sex and strain, was placed in a pillar grid cage and placed in one of the two side chambers, while another identical empty cage was placed on the other side. The apparatus was cleaned with 70% ethanol after each trial. Physical contact with the nose, head and forelimbs around the cage was defined as sniffing behavior (Sniffing). The time mice spent in each chamber (Sociability Chamber Time) and the number of entries into each chamber (Chamber Entries) were analyzed by Noldus Observer software (Ethovision 11.0).

[0051] ②Buried beads experiment Each mouse was placed in a clean standard home cage (27 cm × 16.5 cm × 12.5 cm) with 20 black glass marbles (15 mm diameter) arranged in a symmetrical 4 × 5 cm grid 2–3 cm deep on the top. Investigators counted the number of glass beads buried by the mouse within 30 min, and beads that were covered by >50% of the bead layer were defined as “buried”. Experiments were tested under dim light conditions (-15 lux).

[0052] ③Hair grooming experiment The Noldus Observer software (Ethovision 11.0) was used. Mice were individually placed in clean standard mouse cages, and the camera was placed 15 cm away from the cage. The behavior of the mice was recorded for 20 minutes under low light conditions (40 lux) to facilitate the repetitive self-grooming behavior of the mice. In the last 10 minutes of the test, the mice were manually scored for repetitive self-grooming behavior, and the average cumulative time and the proportion of incorrect transitions were calculated. The grooming behavior microstructure was analyzed using the grooming analysis algorithm (grooming steps: paws, face, body, legs, tail / genitals).

[0053] The results of the three-chamber social experiment test of four groups of mice are as follows Fig.13 As shown, from Fig.13 As can be seen in A, both C57BL / 6-Vehicle mice and C57BL / 6-LP17 mice showed normal sociability in terms of the parameter of indoor time (Sociability Chamber Time), and stayed longer in the room with the novle mouse than in the room with the novle object (Novle Object). The BTBR-Vehicle group mice showed typical sociability defects in terms of indoor time, but this lack of sociability was improved in the BTBR-LP17 mice injected with the TREM1-specific peptide antagonist LP17 in the early developmental stage. Fig.13 B is the number of times the four groups of mice entered the social chamber (Chamber Entries). As can be seen from the figure, there is no significant difference in the number of times the four groups of mice entered the social chamber. Fig.13C is a graph of the sniffing time of the four groups of mice to the novel mouse and the novel object. As can be seen from the graph, the C57BL / 6-Vehicle mice sniffed the novel mouse for a longer time, and the situation was similar for the C57BL / 6-LP17 mice. There was no significant difference in the time the BTBR-Vehicle group mice sniffed the novel mouse and the novel object. However, after the BTBR-LP17 was injected with the TREM1-specific peptide antagonist LP17 at P5 and P6, the time the BTBR mice sniffed the novel mouse was increased, and the time they sniffed the novel mouse was longer than the time they sniffed the novel object, which was a significant difference. Fig.13 D is the number of times the four groups of mice entered the three chambers. It can be seen from the figure that the four groups of mice entered the three chambers the same number of times, and no innate chamber side preference was found.

[0054] from Fig.14 As can be seen in the figure, injection of the TREM1-specific peptide antagonist LP17 in early development reduced marble burial and repetitive self-grooming behaviors in BTBR mice. Fig.14 A, The number of glass beads buried by the two groups of C57BL / 6 mice within 30 minutes was normal and similar, significantly lower than that of BTBR-Vehicle mice, and the number of glass beads buried by BTBR mice decreased after LP17 treatment; Fig.14 As can be seen in B, the two groups of C57BL / 6 mice spent almost the same amount of time on self-grooming, and the BTBR-Vehicle mice spent significantly more time on self-grooming. However, this situation was significantly improved in BTBR mice by injecting LP17.

[0055] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein by equivalents. These modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the embodiments of the present invention, and they should all be included in the scope of the claims and specification of the present invention.

Claims

1. Application of LP17 in the preparation of intervention drugs for the prevention / treatment of autism.

2. The use according to claim 1, characterized in that: The LP17 can inhibit TREM1 expression.

3. The use according to claim 2, characterized in that: The LP17 can reduce central nervous system inflammation.

4. The use according to claim 1, characterized in that: The drug acts on patients with autism in the early postnatal period.

5. The use according to claim 4, characterized in that: The drug is administered by intracranial injection.

6. The use according to claim 5, characterized in that: The effective dosage of the drug is 20 mg / kg.

7. The use according to any one of claims 1 to 6, characterized in that: The drug contains pharmaceutically acceptable excipients.

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