Scorpion oligopeptide and application thereof

Abstract

The application belongs to the technical field of biological medicine, and particularly relates to a Buthus martensii Karsch (Buthus martensii Karsch) oligopeptide and application thereof in preparation of anti-platelet and anti-thrombus drugs or compositions. Buthus martensii The application discloses an anti-platelet oligopeptide discovered by combining an active-oriented separation with computer virtual screening based on a Karsch transcriptome database. The oligopeptide can not only inhibit platelet aggregation, but also effectively inhibit platelet adhesion and expansion. In a thrombus model experiment in vivo, the Buthus martensii Karsch oligopeptide discovered by the application can well inhibit FeCl3-induced carotid artery thrombosis of a mouse. Meanwhile, it is found that the Buthus martensii Karsch oligopeptide does not have obvious bleeding tendency. In conclusion, the Buthus martensii Karsch oligopeptide can be used as an anti-platelet aggregation and anti-thrombus drug to play a certain positive role in the prevention and treatment of thrombotic diseases.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, and specifically relates to a whole scorpion oligopeptide and its applications. Background Technology

[0002] Thrombosis is a common pathological process, primarily caused by abnormal activation of clotting factors and platelets in the blood, leading to blood coagulation and thrombus formation. When blood coagulation is abnormal or occurs in inappropriate locations, thrombi form, causing vascular obstruction and potentially triggering various diseases, even life-threatening ones such as myocardial infarction, stroke, deep vein thrombosis, and pulmonary embolism. Platelets play a crucial role in hemostasis and thrombus formation, making platelet targeting essential for treating thrombotic diseases. Currently used antiplatelet drugs, such as aspirin, clopidogrel, and ticagrelor, directly and irreversibly inhibit platelet activation and aggregation by inhibiting platelet cyclooxygenase and P2Y12 receptors, exhibiting significant antithrombus formation effects. However, they also carry a high risk of bleeding. Therefore, identifying and developing novel targets and drugs for targeted therapy of thrombotic diseases without causing bleeding is of paramount importance.

[0003] Vertebrate lonesome kinase (VLK) is the first secreted protein tyrosine kinase discovered in mammals. After being released from cells, VLK can phosphorylate various proteins located on tyrosine residues in specific functional domains within the secretory pathway and extracellular space. The regulation of physiological functions by endogenous VLK secretion into the extracellular space was first identified in platelets. Studies have found that VLK is highly expressed in platelets, located in secretory granules, and quantitatively released after platelet activation and degranulation upon stimulation by thrombin receptor activating peptide (TRAP). Current research demonstrates that VLK-specific deficiency in mouse megakaryocytes and platelets leads to platelet dysfunction and reduced thrombus formation without prolonging bleeding time, offering the potential of VLK as a novel therapeutic strategy for thrombosis. Extensive literature review has identified three VLK inhibitors: morula-derived ketone C, psoralen isoflavone, and psoralen dihydroflavonoid methyl ether, all of which are small molecule inhibitors. There are currently few VLK inhibitors available, and no peptide inhibitors specifically target VLK. In conclusion, VLK is a promising target for antithrombotic drugs.

[0004] The Chinese medicinal herb *Quanxie* refers to the East Asian scorpion, an animal belonging to the family Scorpionida Buthus martensiiThe dried body of the scorpion (Karsch) possesses the effects of calming wind and relieving spasms, clearing the meridians and relieving pain, and attacking toxins and dispersing nodules. Whole scorpions contain chemical components including proteins, amino acids, lipids, steroids, alkaloids, coumarins, and quinones. Protein content is the highest in whole scorpions and is the main active substance. Current research indicates that purified whole scorpion fluid has antiplatelet activity; however, no antiplatelet oligopeptides derived from whole scorpions have been found to date. Summary of the Invention

[0005] Purpose of the invention: The purpose of this invention is to provide a novel oligopeptide that inhibits platelet function.

[0006] Technical solution: An oligopeptide, characterized in that the oligopeptide is PQ7, and its amino acid sequence is Pro-Asn-Gly-Asn-Leu-Arg-Gln, as shown in SEQ ID NO.3.

[0007] A pharmaceutical composition characterized by comprising the oligopeptide and pharmaceutically acceptable excipients.

[0008] The use of the oligopeptide or the pharmaceutical composition in the preparation of antiplatelet or antithrombotic drugs.

[0009] The use of the oligopeptide or the pharmaceutical composition in the preparation of products that improve microcirculation.

[0010] The use of the oligopeptide or the pharmaceutical composition in the preparation of a medicament for the prevention or treatment of coagulation disorders, thrombosis, or microcirculatory disturbances, characterized in that the diseases are ischemic cardiovascular and cerebrovascular diseases, thromboembolic diseases, peripheral vascular ischemic diseases, diabetic vascular complications, and fundus microcirculatory disturbances.

[0011] The application is characterized in that the diseases include coronary heart disease, angina pectoris, myocardial infarction, cerebral infarction, transient ischemic attack, cerebral ischemia, arteriosclerosis, deep vein thrombosis of the lower extremities, arteriosclerosis obliterans of the lower extremities, thromboangiitis obliterans, diabetic foot, pulmonary embolism, renal artery thrombosis or embolism, retinal artery occlusion, retinal vein occlusion, atrial fibrillation with thromboembolism, post-thrombotic syndrome of deep veins, or peripheral artery thrombotic disease.

[0012] The use of the oligopeptide or the pharmaceutical composition in the preparation of drugs or cosmetics that improve skin microcirculation.

[0013] Specifically: via East Asian pincers Buthus martensiiA theoretical protein database was constructed using the Karsch transcriptome database, and an antiplatelet oligopeptide was obtained by combining activity-directed separation and virtual screening. This oligopeptide inhibits the activity of vertebrate solitary kinase, exhibits antiplatelet activity, and has value in the prevention and treatment of thrombotic diseases. The oligopeptide was derived from the East Asian scorpion. Buthus martensii Karsch transcriptome information, named PQ7; The amino acid sequence of the oligopeptide PQ7 provided by this invention is: Pro-Asn-Gly-Asn-Leu-Arg-Gln.

[0014] The present invention has the following advantages and beneficial effects: The oligopeptide PQ7 provided by this invention has a novel amino acid sequence not reported in the literature. It is the first discovery that PQ7 is a solitary kinase inhibitor in vertebrates. Testing showed that it significantly inhibited thrombin-induced platelet aggregation in a dose-dependent manner. The oligopeptide PQ7 provided by this invention also inhibited platelet expansion at collagen and adhesion and expansion of fibrinogen. Antithrombotic drugs mainly fall into three categories: anticoagulants, antiplatelet drugs, and fibrinolytic drugs. Antiplatelet drugs are the main drugs for the prevention and treatment of thrombotic diseases, such as aspirin and clopidogrel. Their adverse reactions include bleeding phenomena, such as subcutaneous bleeding, gastrointestinal bleeding, and intracranial hemorrhage, which can lead to death or disability. Therefore, antithrombotic drugs cannot be taken long-term for patients with stroke, as this may worsen their condition. Therefore, finding drugs that do not cause bleeding while preventing thrombosis has always been a technical challenge in this field. This invention discovers that the scorpion oligopeptide PQ7 has antithrombotic activity, effectively inhibiting FeCl3-induced carotid artery thrombosis in mice, without a significant risk of bleeding.

[0015] Therefore, this invention provides a new approach and research foundation for the development of novel drugs for antiplatelet and / or prevention and treatment of thrombotic diseases. This oligopeptide has broad application prospects in products that improve microcirculation and in antiplatelet drugs. Attached Figure Description

[0016] Figure 1 The discovery of the active components of whole scorpion water extract; where A represents the inhibitory effect of different molecular weight components of whole scorpion water extract on VLK; and BD represent the effects of different molecular weight components of whole scorpion water extract on thrombin, collagen, and ADP-induced platelet aggregation. Figure 2 The results of analyzing the <3kDa fraction using the East Asian scorpion theoretical protein database and LC-MS are shown in Figure A; A is the UPLC-MS total ion chromatogram of the <3kDa fraction from whole scorpion, and B is the distribution of peptides of different lengths in the <3kDa fraction from whole scorpion. Figure 3The results of virtual screening combined with activity detection to determine the inhibitory effect of scorpion oligopeptides on the activity of vertebrate lonelidomide kinase are shown in the figure; A shows the inhibitory effect of synthesized potential VLK affinity peptides on VLK, B shows the inhibitory activity of PQ7 on VLK, and C shows the three-dimensional molecular docking model of PQ7 and VLK. Figure 4 This is a graph showing the effect of oligopeptide PQ7 on thrombin-induced platelet aggregation; Figure 5 This is a graph showing the effect of oligopeptide PQ7 on platelet adhesion and expansion; A shows the results of platelet adhesion and expansion on collagen as detected by fluorescence microscopy; B shows the expansion area of ​​platelets on collagen; C shows the number of platelets adhering to collagen; D shows the results of platelet adhesion and expansion on fibrinogen as detected by fluorescence microscopy; E shows the expansion area of ​​platelets on fibrinogen; F shows the number of platelets adhering to fibrinogen. Figure 6 The graph shows the effect of scorpion oligopeptide PQ7 on FeCl3-induced carotid artery thrombosis in mice; A is the effect of PQ7 on FeCl3-induced carotid artery blood flow in mice, and B is a statistical graph of blood flow occlusion time. Figure 7 The figure shows the effect of scorpion oligopeptide PQ7 on tail bleeding time in mice; A represents the statistical results of tail bleeding time, B represents the statistical results of bleeding volume, and C represents the observation results of bleeding. Detailed Implementation

[0017] The oligopeptide was synthesized by Nanjing Jietai Biotechnology Co., Ltd.

[0018] Example 1: Discovery of the active components of whole scorpion water extract: Experimental Methods: The aqueous extract of whole scorpion was centrifuged at 5000g at 20℃ using ultrafiltration centrifuge tubes with molecular weight cutoffs of 10kDa and 3kDa. The extract was separated into three fractions: <3kDa, 3-10kDa, and >10kDa. These fractions were then freeze-dried and concentrated, and diluted with water to a crude drug concentration of 1 g / mL. The total extract of whole scorpion (QX), the <3kDa, 3-10kDa, and >10kDa fractions were co-incubated with vertebrate leukopenic kinase (VLK) at a final concentration of 1 mg / mL for 1 h. The inhibitory effect of different fractions on VLK activity was detected using a chemiluminescent kinase activity assay kit to identify the VLK-inhibiting fractions. The QX, <3kDa, 3-10kDa, and >10kDa fractions were added to platelet-rich plasma (PRP) at a final concentration of 1 mg / mL. The effect of the three fractions on platelet aggregation was detected using a platelet aggregation analyzer (Helena Laboratories, USA). The specific method is as follows: Blood was collected from the abdominal aorta of rats using a 3.8% sodium citrate vacuum blood collection tube. The plasma was centrifuged at 70×g for 15 min to collect the upper layer of platelet-rich plasma (PRP). The remaining plasma was centrifuged at 700×g for 15 min to collect the upper layer of platelet-poor plasma (PPP). The PRP was then centrifuged again at 70×g for 10 min to remove as many residual white blood cells and red blood cells as possible. The supernatants were combined and immediately used for platelet aggregation experiments. After preheating the platelet aggregation instrument to 37℃, the optical path of several channels was first checked with double-distilled water, and then the instrument was zeroed using PPP. PRP was incubated with QX, <3kDa, 3-10kDa, and >10kDa (final concentration 1 mg / mL) at 37℃ for 5 min. Then, thrombin, 12.5 μg / mL collagen, or 25 μM ADP at a final concentration of 1.25 U was added to induce platelet aggregation. The stirring speed was set to 600 rpm. Platelet aggregation was recorded using a platelet aggregator for 10 or 5 minutes, including the maximum aggregation rate. Raw data were analyzed using GraphPadPrism 9.0 software. Measurements are expressed as the mean ± standard error of the mean for each independent experiment. One-way ANOVA was used for between-group analysis. p A value <0.05 is considered statistically significant.

[0019] Experimental results: such as Figure 1 As shown in Figure A, the <3kDa fraction of the whole scorpion water extract significantly inhibited the activity of the solitary kinase in vertebrates. Figure 1 As shown in BD, the <3kDa component of the whole scorpion water extract is the key component for inhibiting platelet aggregation.

[0020] Example 2: Analysis of <3kDa components using the East Asian scorpion theoretical protein database combined with LC-MS: Experimental methods: Transcriptome data (SRR17055723, SRR17055724, and SRR17055725) of *Scorpius esculentus* were obtained from the NCBI SRA database (https: / / www.ncbi.nlm.nih.gov / sra). The transcriptome files were converted to Fasta format using Sratoolkit.3.1.0 software, and transcripts were assembled using Trinity (Ver.2.13.2) software and translated using TransDecoder (Ver.5.5.0) software to obtain the theoretical protein database of *Scorpius esculentus*.

[0021] The lyophilized powder of the scorpion fraction with a strength of <3 kDa was sent to Beijing Baitai Parker Biotechnology Co., Ltd. Mass spectrometry was performed using an Easy-nLC1200 / QExactive liquid chromatography-mass spectrometer. Acquisition mode: PRM; Primary mass spectrometry parameters: resolution: 70000; AGC target: 3×10⁻⁶. 6 Maximum IT: 100 ms; Scan range: 100–1500 m / z. Secondary mass spectrometry parameters: Resolution: 17500; AGC target: 2×10⁻⁶ m / z. 5 Maximum IT: 100 ms; NCE: 28%. The collected results and the theoretical protein database of East Asian pincers scorpion were imported into pFind software for comparison to construct a peptide library of the < 3 kDa region of the whole scorpion.

[0022] Experimental results: such as Figure 2 As shown, Figure 2 A shows the UPLC-MS total ion chromatogram of the <3 kDa fraction of whole scorpion. The mass spectrometry data were searched using the open-access library search mode of pFind 3.0 software in conjunction with the transcriptome data from three groups. The conditions were set as PPM ± 10, PSM ≥ 20, and FDR ≤ 1%. After screening, a total of 510 peptides were found across the three transcriptomes, establishing a peptide library containing 510 peptides from the <3 kDa fraction of whole scorpion. Figure 2 B represents the distribution of peptides of different lengths in the scorpion < 3 kDa fraction, with the most abundant peptide being 8-peptide.

[0023] Example 3: Virtual screening combined with activity detection to identify scorpion oligopeptides that inhibit the activity of solitary kinase in vertebrates: Experimental Methods: First, peptides from the peptide library were imported into the ToxinPred website (https: / / webs.iiitd.edu.in / raghava / toxinpred / ) to predict the toxicity of each peptide, and potentially toxic peptides were screened out. Then, VLK was molecularly docked with non-toxic peptides using Schrödinger software. The amino acid sequence of the human VLK protein was downloaded from NCBI, and the VLK protein crystal structure was predicted de novo using RoseTTAFold. The predicted VLK protein crystal structure was imported into Schrödinger software, and these protein molecules were processed, including water molecule removal, hydrogenation, and charge calculation, to select the original ligand-binding region as the docking region. Using VLK protein as the receptor and peptides as ligands, the Peptide Docking module of Schrödinger software was used to screen oligopeptides in the transcriptome-common peptide segment for peptides. Glidescore, an empirical scoring function emphasizing geometric and energy-based rapid screening, was initially selected. Then, peptides with a docking score < -10 were subjected to MM-GBSA calculations. This physics-based energy calculation method combines molecular mechanics (MM) force fields and solvation models to directly calculate the binding free energy based on physicochemical principles. Potential VLK affinity peptides were screened after comprehensive evaluation. The sequences of these affinity peptides were submitted to Nanjing Jietai Biotechnology Co., Ltd. for synthesis. The inhibitory effect of these affinity peptides on VLK activity was detected using a chemiluminescent kinase activity assay kit to identify the inhibitory peptides of the VLK protein. Raw data were analyzed using GraphPadPrism 9.0 software. Measurements are expressed as the mean ± standard error of the mean for each independent experiment. One-way ANOVA was used for inter-group analysis. p A value <0.05 is considered statistically significant.

[0024] Experimental Results: Five potential VLK affinity peptides with MM-GBSA docking score < -11, MM-GBSA dG Bind < -30 kcal / mol, and no toxicity or allergenicity were screened using molecular docking. Table 1 shows the relevant information of the five potential VLK affinity peptides. These five potential affinity peptides were synthesized, and their inhibitory effect on VLK protein was tested. Figure 3 As shown in Figure A, PQ7 and KK6 can significantly inhibit VLK protein activity, with PQ7 showing better inhibitory effects. Figure 3 B represents the inhibitory activity of PQ7 against VLK. The results indicate that PQ7 inhibits VLK activity in a dose-dependent manner, IC50... 50 The value is 209.8 μM. Figure 3C represents the three-dimensional docking model of PQ7 and VLK. PQ7 interacts with the Met-16, Glu-112, Gln-115, Asp-150, Gln-155, Thr-166, and Asp-167 sites of VLK via hydrogen bonds.

[0025] Table 1. Information related to 5 potential VLK affinity peptides .

[0026] Example 4: The inhibitory effect of oligopeptide PQ7 on platelet aggregation: Experimental Methods: Blood was collected from the abdominal aorta of rats using a 3.8% sodium citrate vacuum blood collection tube. The platelet-rich plasma (PRP) layer was collected after centrifugation at 70×g for 15 min. The remaining plasma was centrifuged at 700×g for 15 min, and the anemic platelet-rich plasma (PPP) layer was collected. The PRP was then centrifuged again at 70×g for 10 min to remove as many residual white blood cells and red blood cells as possible. Prostaglandin (final concentration 1 mM) was added, and the platelet-rich plasma was centrifuged again at 700×g for 10 min. The supernatant was discarded, and the platelets were resuspended in calcium-free thiamethoxam solution to obtain washed platelets. Platelet aggregation experiments were immediately performed. A control group, low-, medium-, and high-dose groups, and a positive control group were included. 10 μL of physiological saline, oligopeptide PQ7 (final concentrations of 0.25 mM, 0.5 mM, and 1 mM), aspirin (final concentration of 0.25 mM), and 180 μL of PRP were pre-incubated at 37 ℃ for 5 min to adjust the baseline. After the baseline stabilized, thrombin with a final concentration of 1.25 U was added to induce platelet aggregation. The degree of platelet aggregation was recorded using a platelet aggregator for 10 min, and the maximum aggregation degree of platelets during the aggregation process was recorded. The raw data were analyzed using GraphPadPrism 9.0 software. The measured values ​​are expressed as the mean ± standard error of the mean of each independent experiment. One-way ANOVA was used for intergroup analysis. p A value <0.05 is considered statistically significant.

[0027] Experimental results: such as Figure 4 As shown, the effect of scorpion oligopeptide PQ7 on thrombin-induced platelet aggregation was investigated, and the results showed that oligopeptide PQ7 inhibited platelet aggregation in a dose-dependent manner.

[0028] Example 5: The effect of oligopeptide PQ7 in inhibiting platelet adhesion and spread: Experimental Methods: Place coverslips in a 24-well plate and coat with 250 μL of 5 μg / mL collagen or 50 μg / mL fibrinogen solution overnight at 4 °C. Aspirate the coating solution and wash three times with pre-cooled PBS. Centrifuge fresh PRP at 700×g for 10 min, remove the supernatant, and add Ca-free... 2+After washing three times with Tyrode's-HEPES buffer, the platelets were resuspended in Tyrode's-HEPES buffer to achieve a platelet concentration of 2 × 10⁻⁶. 7 Platelets were washed with 10 μL of oligopeptide PQ7 (final concentration 500 μM) and 180 μL of water and incubated at 37 °C for 10 min. Aspirin (final concentration 250 μM) was used as a positive control. Then, 10 μL of 20 mM CaCl2 solution was added to each well of a 24-well plate coated with collagen or fibrinogen and incubated at 37 °C for 1 h. Unattached platelets were removed by washing three times with PBS. The attached platelets were fixed with 4% (w / v) paraformaldehyde for 10 min, followed by infiltration with 4% (w / v) Triton X-100 for 5 min. After washing three times with PBS, the plates were stained with FITC-labeled phalloidin (final concentration 200 nM) in the dark for 30 min. After washing with PBS, the plates were mounted with anti-fluorescence quenching mounting medium. The plates were observed under a fluorescence microscope, with six fields of view captured randomly, and quantified using ImageJ software. The raw data were analyzed using GraphPadPrism 9.0 software. Measured values ​​are expressed as the mean ± standard error of the mean for each independent experiment. One-way ANOVA was used for between-group analysis. p A value <0.05 is considered statistically significant.

[0029] Experimental Results: During thrombosis, platelet aggregation can be divided into three stages: platelet adhesion, expansion, and stabilization. Platelet adhesion refers to the process where, after endothelial cell damage, circulating platelets adhere to the wound site via the binding of GPVI-IX-V and VWF-collagen complex. This adhesion process initiates platelet expansion, where conformational changes in αIIbβ3 on platelets enhance their affinity for fibrinogen, leading to rapid platelet aggregation.

[0030] Figure 5 AC results showed that, compared with the control group, oligopeptide PQ7 significantly reduced the area of ​​platelet expansion on collagen, but did not affect the amount of platelets adhering to collagen. Figure 5 DF showed that, compared with the control group, the number of platelets adhering to fibrinogen was significantly reduced after treatment with oligopeptide PQ7, and the area of ​​platelet expansion on fibrinogen was significantly reduced.

[0031] Example 6: Scorpion oligopeptide PQ7 inhibits FeCl3-induced carotid artery thrombosis in mice: Experimental Methods: Male ICR mice weighing 20-25g were used as experimental animals. After one week of acclimatization, they were randomly divided into 5 groups (n=6): a blank control group (physiological saline), a positive control group (heparin sodium, HS, 1000 U / kg), and low, medium, and high dose PQ7 groups (5, 10, and 20 mg / kg). All groups received the corresponding drug via tail vein injection. Ten minutes later, the mice were anesthetized. After fixing the mice in a supine position, the neck was prepared and disinfected, and the skin was cut along the midline. The left common carotid artery was bluntly dissected, with a white film placed underneath to enhance imaging contrast. Baseline blood flow was recorded using a Moor FLPI-2 laser speckle imaging system (0 min). 100 μL of 10% (w / v) FeCl3 solution was dripped onto the left common carotid artery of the mouse. After 1 min, the FeCl3 solution was absorbed with a cotton ball, rinsed with a small amount of physiological saline, and residual fluid was aspirated. Carotid artery blood flow was continuously imaged every 1 min, and the occlusion time was recorded. Regardless of whether the artery was occluded, the humane endpoint was defined as 25 minutes after exposure of the carotid artery. Raw data were analyzed using GraphPadPrism 9.0 software. Measurements are expressed as the mean ± standard error of the mean for each independent experiment. One-way ANOVA was used for between-group analysis. p A value <0.05 is considered statistically significant.

[0032] Experimental results: The oligopeptide can inhibit FeCl3-induced carotid artery thrombosis in mice. Figure 6 As shown in Figure A, which illustrates the observation of FeCl3-induced carotid artery thrombosis in mice, and Figure B, which shows the average time to disappearance of blood flow in the model group (6 min), the average time to disappearance of blood flow in the positive control group (heparin sodium) (22 min), and the average times to disappearance of blood flow in the high, medium, and low dose groups of the scorpion oligopeptide (7 min, 12 min, and 17 min, respectively). Compared with the model group, the positive control group and the high and medium dose groups of the scorpion oligopeptide significantly prolonged the time to disappearance of blood flow, indicating that the scorpion oligopeptide exhibits a clear dose-dependent effect.

[0033] Example 7: Scorpion oligopeptide PQ7 showed no significant risk of bleeding. Experimental Methods: Tail amputation experiment. Male ICR mice weighing 20-25g were selected as experimental animals. After one week of acclimatization, the mice were randomly divided into five groups: a blank control group, a positive control group, and high, medium, and low dose groups of the scorpion oligopeptide described in this invention, with six mice in each group. The dosages were as follows: the blank control group received physiological saline; the positive control group received heparin sodium 1000 U / kg; and the high, medium, and low doses of the scorpion oligopeptide were 5 mg / kg, 10 mg / kg, and 20 mg / kg, respectively, administered intravenously. The mice were placed in a tail vein injection device. Ten minutes after administration, a 5 ml centrifuge tube containing 4 ml of physiological saline was prepared. The mouse tail was amputated 2 mm from the tip, and the amputated tail was placed in the centrifuge tube. The bleeding time was recorded from the onset of blood flow until it stopped. The physiological saline was collected after bleeding, the cells were sonicated, centrifuged at 10,000 rpm for 5 minutes, and the supernatant was collected. The absorbance at 540 nm was measured using an ELISA reader to indicate the amount of bleeding. The raw data were analyzed using GraphPadPrism 9.0 software. Measured values ​​are expressed as the mean ± standard error of the mean of each independent experiment. Between-group analysis was performed using one-way ANOVA. p A value <0.05 is considered statistically significant.

[0034] Experimental results: The scorpion oligopeptide PQ7 described in this invention showed no significant bleeding risk at low, medium, and high doses. Figure 7 As shown, the average tail bleeding time in the blank control group was 201 s, and in the positive control group it was 533 s. The average bleeding times in the low, medium, and high dose groups of the compound described in this invention were 232 s, 288 s, and 291 s, respectively. Compared with the blank control group, the heparin sodium group had a significant bleeding risk, while the bleeding time in the low, medium, and high dose groups of the scorpion oligopeptide PQ7 was not significantly different from that in the blank control group. This indicates that the scorpion oligopeptide PQ7 has virtually no effect on the normal physiological hemostasis function of mice and does not pose a significant bleeding risk.

Claims

1. An oligopeptide, characterized in that, The oligopeptide is PQ7, and its amino acid sequence is Pro-Asn-Gly-Asn-Leu-Arg-Gln, as shown in SEQ ID NO.

3.

2. A pharmaceutical composition, characterized in that... It contains the oligopeptide of claim 1 and pharmaceutically acceptable excipients.

3. The use of the oligopeptide according to claim 1 or the pharmaceutical composition according to claim 2 in the preparation of a vertebrate solitary kinase inhibitor drug.

4. The use of the oligopeptide according to claim 1 or the pharmaceutical composition according to claim 2 in the preparation of antiplatelet or antithrombotic drugs.

5. The use of the oligopeptide according to claim 1 or the pharmaceutical composition according to claim 2 in the preparation of products that improve microcirculation.

6. The use of the oligopeptide according to claim 1 or the pharmaceutical composition according to claim 2 in the preparation of a medicament for the prevention or treatment of diseases related to coagulation disorders, thrombosis, or microcirculatory disturbances, characterized in that, The diseases mentioned are ischemic cardiovascular and cerebrovascular diseases, thromboembolic diseases, peripheral vascular ischemic diseases, diabetic vascular complications, and fundus microcirculation disorders.

7. The application according to claim 6, characterized in that, The diseases mentioned include coronary heart disease, angina pectoris, myocardial infarction, cerebral infarction, transient ischemic attack, cerebral ischemia, arteriosclerosis, deep vein thrombosis of the lower extremities, arteriosclerosis obliterans of the lower extremities, thromboangiitis obliterans, diabetic foot, pulmonary embolism, renal artery thrombosis or embolism, retinal artery occlusion, retinal vein occlusion, atrial fibrillation with thromboembolism, post-thrombotic syndrome of deep veins, or peripheral artery thrombotic disease.

8. The use of the oligopeptide according to claim 1 or the pharmaceutical composition according to claim 2 in the preparation of a drug or cosmetic for improving skin microcirculation.

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