Bamboo shoot conus polypeptide Te-SS II and application thereof in analgesia

Abstract

The invention belongs to the technical field of polypeptide preparation, and particularly relates to a bamboo shoot conus polypeptide Te-SS II and application thereof in analgesia. The conus polypeptide toxin Te-SS I with analgesic activity is found from bamboo shoot conus by utilizing high-throughput transcriptomics, then linear peptide is synthesized by adopting a polypeptide solid-phase synthesis method, an oxidized peptide Te-SS II containing one disulfide bond is obtained after the disulfide bond is formed by adopting an air oxidation method, then the oxidized peptide Te-SS II is purified by adopting high performance liquid chromatography, and mass spectrum identification is performed. The Te-SS II has one disulfide bond formed by two cysteines and has an efficient analgesic effect on zebra fish and mice. Therefore, the bamboo shoot conus polypeptide Te-SS II provided by the invention can be used for preparing analgesic products, and a new breakthrough is brought to the field of pain treatment.

Description

Technical Field

[0001] The invention belongs to the technical field of polypeptide preparation, and particularly relates to a bamboo shoot and cone snail polypeptide Te-SSⅡ and an application thereof in analgesia. Background Art

[0002] Pain, a complex sensory and emotional response to potential or actual injury, is a common symptom in clinical practice. Opioids have long been the mainstay of analgesic therapy, but their addictiveness, tolerance, and side effects limit their clinical application. Therefore, the search for new analgesics has become a crucial area of medical research, and the diversity of marine life offers an underexplored treasure trove for drug discovery.

[0003] Cone snails, also known as cone snails, are gastropod molluscs with approximately 500 species worldwide, found in warm seas. China is home to over 80 species of cone snails. Conotoxins (CTx) are a class of active peptides secreted by cone snails, primarily used for prey capture and defense. Conotoxins specifically target multiple receptors, including nicotinic acetylcholine receptors, voltage-gated ion channels, and G protein-coupled receptors. They hold great promise for the treatment of diverse and challenging conditions, including neuralgia, addiction, epilepsy, and cancer.

[0004] In recent years, studies have revealed that certain types of cone snail toxins exhibit significant analgesic potential. They act by blocking pain signal transmission through specific neurotransmitter receptors, such as N-type calcium channels or sodium channels, demonstrating an analgesic mechanism distinct from that of traditional opioids. The discovery of these toxins offers new insights into the development of novel non-opioid analgesics. However, there have been no reports on the bamboo shoot cone snail peptide Te-SSⅡ. Summary of the Invention

[0005] The purpose of the present invention is to provide a bamboo shoot cone snail polypeptide Te-SSⅡ and its application in analgesia, which has a highly effective analgesic effect.

[0006] The present invention provides a bamboo shoot cone snail polypeptide Te-SSⅡ. The amino acid sequence of the bamboo shoot cone snail polypeptide Te-SSⅡ is shown in SEQ ID NO: 1.

[0007] The present invention provides a preparation method of the bamboo shoot cone snail polypeptide Te-SSⅡ described in the above technical scheme, wherein a linear peptide is synthesized by a polypeptide solid-phase synthesis method, and an oxidized peptide Te-SSⅡ containing one disulfide bond is obtained after forming a disulfide bond by an air oxidation method, and then purified by high-performance liquid chromatography and then identified by mass spectrometry.

[0008] Preferably, the specific steps of the air oxidation method include:

[0009] The purified linear peptide was placed in a tris(hydroxymethylaminomethane) hydrochloride aqueous buffer system for oxidation to obtain the oxidized peptide Te-SSⅡ.

[0010] The tris(hydroxymethylaminomethane) hydrochloride aqueous solution buffer system includes substances with the following concentrations: 0.1 mol / L Tris-HCl, 1 mmol / L ethylenediaminetetraacetic acid, 1 mmol / L reduced glutathione, and 1 mmol / L oxidized glutathione.

[0011] Preferably, the mass-to-volume ratio of the purified linear peptide to the tris(hydroxymethyl)aminomethane hydrochloride aqueous solution buffer system is 0.5 mg:1 mL.

[0012] Preferably, the oxidation treatment is accompanied by stirring; the oxidation treatment time is 24 hours; and the oxidation treatment temperature is 15-25°C.

[0013] The present invention provides the use of the bamboo shoot cone snail polypeptide Te-SSⅡ described in the above technical solution in the preparation of analgesic products.

[0014] Preferably, the product includes: medicine.

[0015] Preferably, the types of analgesia include: one or more of acute thermal pain, inflammatory pain and neuralgia.

[0016] The present invention provides a medicine for analgesia, which comprises: the bamboo shoot and cone snail polypeptide Te-SSⅡ described in the above technical solution.

[0017] Preferably, the final concentration of the bamboo shoot cone snail polypeptide Te-SSⅡ in the drug is: 0.625-10 μM.

[0018] Beneficial effects:

[0019] The present invention provides a bamboo shoot cone snail polypeptide Te-SSⅡ, the amino acid sequence of the bamboo shoot cone snail polypeptide Te-SSⅡ is shown in SEQ ID NO: 1, and the specific sequence is: QDGAQICFWKICPPSPW. The present invention discovered the cone snail polypeptide toxin Te-SSⅠ with analgesic activity from the bamboo shoot cone snail (Conus telebra) by using high-throughput transcriptomics, and then synthesized a linear peptide by using peptide solid phase synthesis (SPPS). After air oxidation to form a disulfide bond, an oxidized peptide Te-SSⅡ containing one disulfide bond was obtained. After purification by high-performance liquid chromatography and mass spectrometry identification, Te-SSⅡ with two cysteines forming one disulfide bond and having a highly effective analgesic effect on zebrafish and mice was obtained. Therefore, the bamboo shoot cone snail polypeptide Te-SSⅡ provided by the present invention can be used to prepare analgesic products, bringing new breakthroughs in the field of pain treatment. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0021] Figure 1 The results of HPLC analysis of the linear peptide Te-SSⅡ provided by the present invention;

[0022] Figure 2 The mass spectrometry identification results of the linear peptide Te-SSⅡ provided by the present invention;

[0023] Figure 3 The results of HPLC analysis of the oxidized peptide Te-SSⅡ provided by the present invention;

[0024] Figure 4 The mass spectrometry identification results of the oxidized peptide Te-SSⅡ provided by the present invention;

[0025] Figure 5 The effect of the polypeptide Te-SSⅡ provided by the present invention on the movement trajectory and total movement distance of zebrafish larvae;

[0026] Figure 6 This is the concentration-response curve after administration of different concentrations of cone snail toxin Te-SSⅡ in the mouse hot plate model provided by the present invention;

[0027] Figure 7 The area under the curve of the analgesic effect in the mouse hot plate model provided by the present invention;

[0028] Figure 8 This is the concentration-response curve after administration of different concentrations of cone snail toxin Te-SSⅡ in the mouse tail-flick model provided by the present invention;

[0029] Figure 9 The area under the curve of the analgesic effect in the mouse tail-flick model provided by the present invention;

[0030] Figure 10 This is the concentration-response curve after administration of different concentrations of cone snail toxin Te-SSⅡ in the mouse CFA model provided by the present invention;

[0031] Figure 11 The area under the curve of the analgesic effect in the mouse CFA model provided by the present invention;

[0032] Figure 12 This is the concentration-response curve after administration of different concentrations of cone snail toxin Te-SSⅡ in the mouse CCI model provided by the present invention;

[0033] Figure 13 The area under the curve of the analgesic effect in the mouse CCI model provided by the present invention;

[0034] Figure 14 The present invention provides the activation effect of Te-SSⅡ on five subtypes of SSTR. DETAILED DESCRIPTION

[0035] In the present invention, unless otherwise specified, the reagents, methods and equipment used are conventionally selected.

[0036] In order to further illustrate the present invention, the solutions provided by the present invention are described in detail below with reference to the accompanying drawings and embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0037] Experimental Description

[0038] 1 Materials and Methods

[0039] 1.1 Experimental Materials

[0040] Chromatographic grade trifluoroacetic acid (TFA) and chromatographic grade acetonitrile (ACN) were purchased from Shanghai Aladdin Biochemical Technology Co., Ltd.; a Waters analytical C18 column (5 μm, 4.6 mm × 250 mm) was purchased from Waters, USA; and an Elite preparative C18 column (10 μm, 10 mm × 250 mm) was purchased from Dalian Elite Analytical Instrument Co., Ltd.

[0041] 1.2 Experimental instruments

[0042] CEM fully automatic microwave peptide synthesizer (LibertyBlue, USA); reversed-phase high-performance liquid chromatography (ThermoFisher, Germany); triple quadrupole liquid chromatography-mass spectrometry (Shimadzu, Japan); desktop freeze dryer (Saifei, China); zebrafish behavioral trajectory tracking system (Shanghai Yusen Biotechnology Co., Ltd.); intelligent hot plate apparatus (Anhui Zhenghua Biological Instrument Equipment Co., Ltd.); rat tail light analgesia (Shanghai Xinruan Information Technology Co., Ltd.); rat and mouse plantar mechanical prick instrument (Anhui Yaokun Biotechnology Co., Ltd.).

[0043] 1.3 Data Processing

[0044] The data were statistically analyzed and processed using GraphPad Prism 8 software. The data between the control group and the experimental group were analyzed using t-test. * indicates p < 0.05, which is statistically different; ** indicates p < 0.01, which is statistically significant; *** indicates p < 0.001, which is highly statistically significant; **** indicates p < 0.0001, which is extremely statistically significant.

[0045] Example 1 Synthesis and Oxidative Folding of Conus Polypeptide Toxin Te-SSⅡ

[0046] (1) Weigh 3 g of 2-Cl resin (degree of substitution Sd = 0.2 mmol / g) and add it to a reactor. Soak it in DCM (20 mL) for 5 min, wash it twice with DMF (20 mL), and wash it once with DCM for the third time. Add 0.8 mmol of the first amino acid at the carboxyl terminal, 12 mL of DCM, and 2 mL of DIEA. React for 90 min. After 90 min, add 4 mL of analytical methanol and 10 mL of DCM, and block the reaction for 30 min.

[0047] (2) Wash with DMF 4 times, add piperidine (20% piperidine + 80% DMF) to remove Fmoc for 20 minutes. Wash the resin with DMF 5 times, take a small amount of resin (10-20 grains) and add it to a test tube. Add 2 drops of ninhydrin (5g / 100mL analytical ethanol) and 2 drops of pyridine, heat at 100℃ for 2 minutes, and color is developed.

[0048] (3) Weigh the next amino acid from the C-terminus to the N-terminus (1.8 mmol) + HOBT (1.8 mmol), add them to the reactor, pour in 10 mL of DMF, add 2 mL of DIC, react for 1 hour, and then wash with DMF four times. Take a small amount of resin for detection, and it is colorless.

[0049] (4) Wash with DMF 4 times, add piperidine (20% piperidine + 80% DMF) to remove Fmoc for 20 minutes. Wash the resin with DMF 5 times, take a small amount of resin (10-20 grains) and add it to a test tube. Add 2 drops of ninhydrin (5g / 100mL analytical ethanol) and 2 drops of pyridine, heat at 100℃ for 2 minutes, and color is developed.

[0050] (5) Repeat steps (3) and (4) until the peptide chain is complete.

[0051] (6) Drain the resin with methanol and place it in a 50 mL centrifuge tube. Add 40 mL of cutting solution (95% TFA + 1% H2O + 2% DODT + 2% TIPS) and shake for 2 h. Finally, divide the cutting solution (filtering the resin) equally into 4 new 50 mL centrifuge tubes, add 40 mL of ice ether, shake evenly, and centrifuge at 3000 rpm for 2 min. The polypeptide remains at the bottom and the upper ether is poured out to obtain the crude polypeptide.

[0052] (7) The polypeptide is separated and purified by HPLC to obtain pure polypeptide, which is then freeze-dried into powder using a vacuum freeze dryer.

[0053] (8) Prepare a tris-HCl aqueous buffer system consisting of 0.1 M Tris-HCl (pH = 6.8), 1 mM ethylenediaminetetraacetic acid (EDTA), 1 mM reduced glutathione (GSH), and 1 mM oxidized glutathione (GSSG). Dissolve the purified linear peptide in the above buffer system at a concentration of 0.5 mg / mL, place it at room temperature, and stir it in air for 24 hours to promote the formation of a disulfide bond between the two free cysteine residues. Subsequently, use preparative RP-HPLC to separate and purify the oxidative folding product. The purified fractions were tested for purity by analytical RP-HPLC, and the molecular weight was identified by LC-MS. The fractions with a purity of more than 95% were screened out, combined, freeze-dried, and sealed in a -80°C refrigerator for storage.

[0054] The linear peptide was synthesized by solid phase peptide synthesis (SPPS) and purified by HPLC and identified by mass spectrometry. Figure 1-Figure 2 As shown, the molecular weight of the linear peptide Te-SSⅡ is 1976.0416 Da.

[0055] The synthesized linear peptide was oxidatively folded by air oxidation, purified by HPLC and identified by mass spectrometry. Figure 3 and Figure 4 The molecular weight of the oxidized peptide Te-SSⅡ is 1975.0280Da.

[0056] It can be seen that the molecular weight of the linear peptide Te-SSⅡ is 1976.0416Da, and the molecular weight of the oxidized peptide Te-SSⅡ is 1975.0280Da, and the difference between the two is about 2Da (two hydrogen atoms), which proves that the disulfide bond is correctly formed.

[0057] Example 2 Pain behavior experiment of zebrafish larvae

[0058] The freeze-dried Te-SSⅡ oxidized peptide powder prepared in Example 1 was weighed and prepared into peptide solutions with final concentrations of 10 μM, 5 μM, 2.5 μM, 1.25 μM and 0.625 μM, respectively, using fish farming water as a solvent.

[0059] Zebrafish larvae at 5 days post fertilization (dpf) were selected and placed in a 24-well plate, with one larva placed in each well, and 8 to 10 zebrafish larvae were set up in each group;

[0060] Experimental group setup: 1 mL of the five concentrations of Te-SSⅡ peptide solution (10 μM, 5 μM, 2.5 μM, 1.25 μM, and 0.625 μM) was added to each well; a tramadol solution with a final concentration of 1.25 μM was added to the positive control group. Separately, two groups of zebrafish larvae were added to the fish culture water, one of which served as the model group (to be subsequently treated with acetic acid at a final concentration of 0.01% in a volume of 1 mL) and the other as the blank control group.

[0061] The juveniles in each of the above treatment groups were left to stand for 10 minutes to adapt to the environment, and then the zebrafish behavior trajectory tracking system was used to record the movement trajectory and speed within 10 minutes, and the zebrafish juveniles with a speed between 0.1 and 2 mm / s were screened for subsequent experiments. After 10 minutes, an acetic acid aqueous solution with a final concentration of 0.01% was added to the wells of each group (the blank control group was added with an equal amount of fish water), and then the movement trajectory and speed of the zebrafish juveniles were continuously monitored within 20 minutes. The movement trajectory and speed were recorded every 2 minutes, for a total of 10 times, and the total movement distance of each zebrafish juvenile in different time periods was calculated. The results are shown in Figure 5 (exist Figure 5 Figure 2 (A) shows the trajectory of movement behavior; the instantaneous speed was detected and displayed in different colors (black, <2 mm / s; green, 2-8 mm / s; red, >8 mm / s); Figure 2 (B) shows the statistics of total movement distance (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001).

[0062] Combine Figure 5The behavioral trajectory diagram shows that, compared with the blank control group, the addition of 0.01% acetic acid significantly enhanced the movement behavior of the zebrafish larvae and accelerated their speed, with a significant increase in the total movement distance, indicating that the acetic acid-induced pain model was successfully established. Combined with the total movement distance diagram, it can be seen that Te-SSⅡ oxidized peptide has a certain regulatory effect on the pain-related movement behavior of zebrafish larvae. With the increase in the concentration of Te-SSⅡ oxidized peptide, the total movement distance of the zebrafish larvae gradually decreased, showing a dose-dependent effect. At the highest tested concentration (10μM), the total movement distance of the zebrafish larvae was significantly lower than that of the pain model group. Therefore, Te-SSⅡ oxidized peptide has a significant analgesic effect on the pain behavioral model of zebrafish larvae.

[0063] Example 3 Pain behavior experiment in mice

[0064] A pain model was established using female mice (18-22 g) to evaluate the analgesic effect of the Te-SSⅡ oxidized peptide from cone snail toxin. The freeze-dried Te-SSⅡ oxidized peptide powder prepared in Example 1 was used as a solvent, and peptide solutions with doses of 20 mg / kg, 10 mg / kg, 5 mg / kg, 2.5 mg / kg, and 1.25 mg / kg were prepared using 0.9% saline as a solvent.

[0065] Experimental setup: The above five dose groups (20 mg / kg, 10 mg / kg, 5 mg / kg, 2.5 mg / kg and 1.25 mg / kg), a 0.9% saline control group and a tramadol positive control group (50 mg / kg) were used as the treatment solutions. Six female mice were selected from each treatment group, and each mouse was intraperitoneally injected (injection dose 10 μL / g) with each treatment solution.

[0066] Acute thermal pain was assessed using the hot plate (55°C paw licking response) and tail flick (50W laser tail flick response) models, chronic inflammatory pain induced by CFA (20 μL CFA was injected into the paw, and mechanical pain threshold was measured by von Frey 3 days later), and the CCI neuralgia model (quadruple ligation of the sciatic nerve, behavioral verification 5 days after surgery) was used to evaluate drug efficacy. Six mice were set up in each model (reference for model establishment: Somatostatin venom analogs evolved by fish-hunting cone snails: From prey capture behavior to identifying drug leads.). Pain thresholds were continuously monitored 1 to 4 hours after drug administration. The success criteria of the model included: foot redness and swelling / decreased pain threshold in the CFA group, and lower limb movement disorders / hyperalgesia in the CCI group. All experiments used the average of two measurements for pain threshold. Among them, the behavioral results of pain in the mouse hot plate model are shown in Figure 6 and Figure 7; The results of pain behavior in the mouse tail-flick model are shown in Figure 8 and Figure 9 ; The results of pain behavior of mouse CFA model are shown in Figure 10 and Figure 11 ; The results of pain behavior in the mouse CCI model are shown in Figure 12 and Figure 13 ;exist Figure 6-Figure 13 One-way ANOVA was used to compare the experimental and model groups. Data are presented as mean ± standard error of the mean (SEM) (n = 6), *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

[0067] The results of the mouse hot plate model experiment showed that Te-SSⅡ reached its maximum analgesic effect 2 hours after administration and the effect lasted for 3 hours ( Figure 6 ). 20mg / kg of Te-SSⅡ increased the pain threshold of mice at 2h more than 50mg / kg of tramadol during the same period, and its area under the curve was equivalent to that of 50mg / kg tramadol ( Figure 7 ), showing a longer-lasting analgesic effect. Te-SSⅡ showed a very significant analgesic effect (p<0.0001) at a moderate dose (5mg / kg), and was dose-dependent. The results of the mouse tail-flick model were basically consistent with the hot plate model ( Figure 8 and Figure 9 ).

[0068] The complete Freund's adjuvant (CFA) pain model showed that Te-SSⅡ had a significant alleviating effect on CFA-induced pain, and the difference was extremely significant compared with the blank control group (p<0.0001, Figure 10 and Figure 11 ). Among them, the area under the analgesic effect curve of 20mg / kg Te-SSⅡ exceeded that of 50mg / kg tramadol, showing a significant analgesic effect. One hour after administration, the increase in the mechanical pain threshold of mice by Te-SSⅡ was close to that of tramadol, and in the following 2 hours, its concentration-response curve was always higher than that of tramadol, showing a long-lasting and potent analgesic effect. At a higher dose (10mg / kg), Te-SSⅡ showed an extremely significant analgesic effect (p<0.0001), and it was obviously dose-dependent. The results of the chronic sciatic nerve constriction injury (CCI) model showed that Te-SSⅡ exhibited significant analgesic activity, and exhibited a stable and long-lasting analgesic effect within 4 hours after administration at a dose of 20mg / kg ( Figure 12 and Figure 13 ).

[0069] Example 4 cAMP cell function experiment

[0070] (1) Cell culture and reagent preparation: The cell line was Flp-In-CHO-SSTR clone #17;

[0071] Complete culture medium: Ham's F-12K medium, 10% fetal bovine serum, 1× penicillin-streptomycin solution, and 600 μg / mL hygromycin B;

[0072] Cell culture medium: Ham's F-12K medium and 10% fetal bovine serum;

[0073] Assay buffer: 1× Hank's balanced salt solution (HBSS), 20 mM 4-hydroxyethylpiperazineethanesulfonic acid (HEPES), 0.1% bovine serum albumin (BSA), and 500 μM 3-isobutyl-1-methylxanthine (IBMX).

[0074] (2) Agonist Activity Assay: Flp-In-CHO-SSTR clone #17 cells were cultured in complete medium at 37°C, 5% CO2, and maintained at near-confluence. 7K cells / well were seeded into a 384-well cell culture plate containing cell culture medium, with 25 μL per well, and incubated overnight at 37°C, 5% CO2.

[0075] Remove the cell culture plate, remove the culture medium, and quickly add 15 μL of detection buffer to the experimental wells. Prepare 8x cone snail toxin polypeptide sample (i.e., Te-SSⅡ prepared in Example 1, initial test concentration is 10 μM) and forskolin (4 μM) in the detection buffer, add 2.5 μL of 8x sample working solution to the cell culture plate, incubate at 37°C for 10 minutes, and then add 2.5 μL of 8x forskolin working solution to the cell culture plate and incubate at 37°C for 30 minutes.

[0076] use Ultra cAMP Assay Kit: Dilute the cAMP tracer labeled with europium chelate and the cAMP-specific monoclonal antibody labeled with Ulight dye in cAMP assay buffer. Add 10 μL of Eu-cAMP tracer to the cell culture plate, followed by 10 μL of Ulight-anti-cAMP. Add 10 μL of assay buffer to the negative control group of the cell culture plate. Incubate at room temperature for 1 hour, and then read the absorbance at 665 nm and 615 nm on an Envision 2105 multi-function microplate reader. Figure 14 .

[0077] Depend on Figure 14As can be seen, SSTR4 mediates effective analgesic and anti-inflammatory effects, while Te-SSII does not activate SSTR1. Its EC50 values for SSTR2, SSTR3, SSTR4, and SSTR5 are 1877nM, 5626nM, 223.3nM, and 3316nM, respectively. This indicates that Te-SSII not only activates SSTR4 but also exhibits weak activation effects on SSTR2, SSTR3, and SSTR5. However, its EC50 values are significantly higher than those for SSTR4, indicating that it still has good selectivity.

[0078] Although the above embodiment provides a detailed description of the present invention, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the scope of protection of the present invention.

Claims

1. A bamboo shoot cone snail polypeptide Te-SSⅡ, characterized in that: The amino acid sequence of the bamboo shoot cone snail polypeptide Te-SSⅡ is shown in SEQ ID NO:

1.

2. The method for preparing the bamboo shoot cone snail polypeptide Te-SSⅡ according to claim 1, characterized in that: The linear peptide was synthesized by solid-phase synthesis. The oxidized peptide Te-SSⅡ containing one disulfide bond was obtained by air oxidation to form disulfide bonds. The peptide was then purified by high performance liquid chromatography and identified by mass spectrometry.

3. The preparation method according to claim 2, wherein the specific steps of the air oxidation method include: The purified linear peptide was placed in a tris(hydroxymethylaminomethane) hydrochloride aqueous buffer system for oxidation to obtain the oxidized peptide Te-SSⅡ. The tris(hydroxymethylaminomethane) hydrochloride aqueous solution buffer system includes substances with the following concentrations: 0.1 mol / L Tris-HCl, 1 mmol / L ethylenediaminetetraacetic acid, 1 mmol / L reduced glutathione, and 1 mmol / L oxidized glutathione.

4. The preparation method according to claim 3, characterized in that The mass volume ratio of the purified linear peptide to the tris(hydroxymethyl)aminomethane hydrochloride aqueous solution buffer system is 0.5 mg:1 mL.

5. The preparation method according to claim 3, characterized in that The oxidation treatment is accompanied by stirring; the oxidation treatment time is 24 hours; and the oxidation treatment temperature is 15-25°C.

6. Use of the bamboo shoot and cone snail polypeptide Te-SSⅡ according to claim 1 in the preparation of analgesic products.

7. The use according to claim 6, characterized in that The products include: medicines.

8. The use according to claim 6, characterized in that The types of analgesia include: one or more of acute heat pain, inflammatory pain and neuralgia.

9. A drug for analgesia, characterized in that The medicine comprises: the bamboo shoot cone snail polypeptide Te-SSⅡ according to claim 1.

10. The drug according to claim 9, characterized in that The final concentration of the bamboo shoot cone snail polypeptide Te-SSⅡ in the medicine is 0.625-10 μM.

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