Maca Pi scorpion polypeptide and application thereof

Abstract

The invention belongs to the technical field of biological medicines, and particularly discloses a Pi scorpion polypeptide and application thereof. Through molecular biology, site-specific mutagenesis, chemical synthesis and other methods, the obtained Pi scorpion polypeptide can effectively inhibit the growth of escherichia coli, pseudomonas aeruginosa, staphylococcus aureus, propionibacterium acnes and related drug-resistant bacteria at low concentration. The pemai Pi scorpion polypeptide has a very good effect in treating or preventing folliculitis, furuncle, impetigo, acne and whelk caused by escherichia coli, pseudomonas aeruginosa, acinetobacter baumannii, propionibacterium acnes, staphylococcus aureus, related drug-resistant bacteria and the like. The pemai Pi scorpion polypeptide disclosed by the invention has good activity in resisting escherichia coli, pseudomonas aeruginosa, acinetobacter baumannii, propionibacterium acnes, staphylococcus aureus and related drug-resistant bacteria, and can be further developed and utilized as an antibacterial drug for treating folliculitis, furuncle, impetigo, acne, whelk and the like.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to Mars scorpion polypeptide and its uses. Background Technology

[0002] Follicular pustulosis is a superficial inflammation of the hair follicle. The pathogen is most commonly Staphylococcus aureus, but sometimes Staphylococcus epidermidis. Folliculitis initially presents as a red, firm papule consistent with the hair follicle opening, or it may begin as a pustular follicle and then rapidly develop into a papular pustule. Folliculitis can also recur and persist for many years; some cases can even develop into deeper infections, forming boils, carbuncles, etc. Follicular pustulosis and other skin diseases are closely related to bacterial infection.

[0003] Acne, pimples, and blackheads are common and frequently occurring diseases among teenagers, and can also occur in middle-aged individuals. Clinical manifestations include whiteheads, blackheads, inflammatory papules, secondary pustules, cysts or nodules, pain, and itching. If infected, they can leave scars, affecting appearance and mental and physical health. Acne is a chronic disease of the sebaceous glands, which can be divided into inflammatory and non-inflammatory acne. It is the second most common dermatological disease, accounting for approximately 7-10% of all skin diseases, affecting individuals aged 11-30, and can occur in both men and women. Treatment methods mainly involve topical medications, oral medications, or a combination of both, and physical therapy is also used. However, overall treatment effectiveness is not ideal, with many adverse reactions, skin damage, or minimal efficacy. Many pathogenic factors and processes are now known to play a key role in the occurrence, development, and outcome of acne, pimples, and blackheads. The most important reason for acne is the role of microorganisms in the pilosebaceous unit, primarily anaerobic Propionibacterium acnes, followed by aerobic Epidermal and Staphylococcus aureus pyogenic bacteria. Some also believe that yeasts are involved in the pathogenesis. Drugs are the main treatment for acne, and anti-acne drugs are a hot topic in dermatological drug research both domestically and internationally. Their mechanisms of action mainly revolve around and target the aforementioned pathogenesis links. Anti-acne drugs are administered via systemic and topical routes. Topical treatment is mainly for mild and moderate acne, while severe acne requires topical treatment in conjunction with a comprehensive therapy. Currently available acne medications are mostly antibiotics, including erythromycin, clindamycin (including phosphate and hydrochloride), and chloramphenicol, which primarily exert antibacterial effects.

[0004] With the widespread use of antibiotics, bacterial resistance has become a serious challenge to global public health. Traditional antibiotics often act on single targets, making them easily evaded by bacteria through gene mutations. In contrast, cationic antimicrobial peptides primarily kill bacteria through a physical mechanism of electrostatic adsorption and disruption of the bacterial cell membrane, making them less likely to induce resistance and considered an important direction for developing next-generation anti-infective drugs.

[0005] Therefore, there is an urgent need to discover more effective anti-infective drugs from natural resources to overcome bacterial resistance. Summary of the Invention

[0006] The purpose of this invention is to address the aforementioned shortcomings of the prior art by providing a Mars-like scorpion polypeptide and its uses. This Mars-like scorpion polypeptide exhibits significant activity against *Escherichia coli*, *Pseudomonas aeruginosa*, *Acinetobacter baumannii*, *Propionibacterium acnes*, *Staphylococcus aureus*, and related drug-resistant bacteria.

[0007] To achieve the above objectives, the present invention adopts the following technical solution: The first object of the present invention is to provide an isolated *Scorpionus martensii* polypeptide or a pharmaceutically acceptable salt thereof, said *Scorpionus martensii* polypeptide comprising the sequence SEQ ID NO:1 or a variant thereof, said variant sequence comprising the amino acid sequence of general formula I or general formula II. The general formula I is X1ILX2X3LWX4X5VX6SIF, where X1, X2, X3, X4, X5, and X6 are amino acid substitution sites, corresponding to positions 1, 4, 5, 8, 9, and 11 of SEQ ID NO:1, and the amino acids used for substitution are basic amino acids or nonpolar amino acids. The general formula II is a sequence formed by deleting 1 to 5 amino acids from general formula I, wherein the deleted amino acids correspond to the sites of the original nonpolar amino acids in SEQ ID NO:1; and the Marsala scorpion polypeptide has the activity of inhibiting Gram-negative bacteria and / or Gram-positive bacteria.

[0008] Furthermore, the general formula II is selected from structures formed by the deletion of one or more of the following amino acids: X1Y2Y3X2X3LWX4X5VX6Y 12 Y 13 Y 14 ,in, X1, X3, X4, and X6 are sites where only amino acid substitution occurs, corresponding to SEQ ID NO:2, and are selected from basic amino acids or nonpolar amino acids, respectively. X2, X5, Y 12 The sites where amino acid substitution or deletion occurs, corresponding to SEQ ID NO:2, are selected from basic amino acids, nonpolar amino acids, or are absent, respectively; Y2, Y3, Y 13 Y 14 For sites where only amino acid deletion occurs, Y2 is selected from isoleucine or is absent, Y3 is selected from leucine or is absent, Y... 13 Selected from leucine or absent, Y 14 Choose phenylalanine or none; Furthermore, the total number of missing amino acids in Formula II does not exceed 5.

[0009] Furthermore, the basic amino acid is selected from arginine, lysine, or histidine; the nonpolar amino acid is selected from alanine, valine, leucine, isoleucine, phenylalanine, tryptophan, or methionine.

[0010] Furthermore, in the general formula I: X1, X3, X4 and X6 are selected from arginine, lysine or histidine; X2 and X5 are selected from valine, glycine, or threonine.

[0011] Furthermore, the amino acid sequence of the Mars scorpion polypeptide is selected from SEQ ID NO:2 to SEQ ID NO:45.

[0012] A second object of the present invention is to provide an isolated nucleic acid molecule comprising a nucleotide sequence encoding the aforementioned Marsella scorpion polypeptide.

[0013] A third object of the present invention is to provide a pharmaceutical composition comprising an effective dose of the above-described antimicrobial peptide and a pharmaceutically acceptable carrier.

[0014] A fourth objective of this invention is to provide the use of the above-mentioned Mars scorpion polypeptide in the preparation of a medicament for treating or preventing bacterial infections, wherein the bacteria include Escherichia coli, Pseudomonas aeruginosa, Acinetobacter baumannii, Propionibacterium acnes, Staphylococcus aureus, and related drug-resistant bacteria.

[0015] Furthermore, the drug includes medications for treating or preventing acne, pimples, and blackheads.

[0016] The fifth objective of this invention is to provide a topical gel formulation, which is prepared from the Mars scorpion polypeptide provided by this invention and a gel matrix, wherein the gel matrix is ​​hydroxymethyl cellulose.

[0017] Compared with existing technologies, the beneficial effects of the technical solution provided by this invention are as follows: (1) The Mars scorpion polypeptide provided by the present invention contains the sequence SEQ ID NO:2 or a variant sequence thereof, the variant sequence containing the amino acid sequence of general formula I or general formula II. Experiments have shown that it has significant inhibitory activity against Escherichia coli, Pseudomonas aeruginosa, Acinetobacter baumannii, Propionibacterium acnes, Staphylococcus aureus and their related drug-resistant bacteria, and that its mechanism is to exert its effect by destroying the cell membrane of bacteria, and it is not easy to produce drug resistance.

[0018] (2) The present invention provides a topical gel preparation, which is formulated from the Mars scorpion polypeptide and gel matrix provided by the present invention. In human efficacy trials, the results show that it is effective in treating acne and has significant therapeutic effects. Attached Figure Description

[0019] Figure 1Image of the venom glands of the Mars scorpion; Figure 2 The isolated wild-type Scorpion Marsips polypeptide gene SmarAMP286; Figure 3 Chromatogram of peptides from wild-type Scorpion Marseilles separated by high performance liquid chromatography; Figure 4 Mass spectra used to identify the molecular weight of polypeptides from wild-type Mars scorpion. Detailed Implementation

[0020] To make the objectives, technical solutions, and advantages of this invention clearer, the specific embodiments and accompanying drawings are described in further detail below. Where specific techniques or conditions are not specified in the embodiments, they are performed according to the techniques or conditions described in the literature in this field or according to the product instructions. Reagents or instruments whose manufacturers are not specified are all commercially available conventional products.

[0021] Definitions and explanations of terms in this invention: The term "isolated" includes naturally occurring, gene expression products, and synthetic peptides.

[0022] In this invention, amino acid sequences or their variants are represented by single-letter symbols. These single-letter and three-letter symbols are well known to those skilled in the art and have the following meanings: A (Ala) is alanine, C (Cys) is cysteine, D (Asp) is aspartic acid, E (Glu) is glutamic acid, F (Phe) is phenylalanine, G (Gly) is glycine, H (His) is histidine, I (Ile) is isoleucine, K (Lys) is lysine, L (Leu) is leucine, M (Met) is methionine, N (Asn) is asparagine, P (Pro) is proline, Q (Gln) is glutamine, R (Arg) is arginine, S (Ser) is serine, T (Thr) is threonine, V (Val) is valine, W (Trp) is tryptophan, and Y (Tyr) is tyrosine.

[0023] Antimicrobial peptides are polypeptides that have the activity of inhibiting or killing microorganisms (such as bacteria, fungi, viruses, etc.).

[0024] Nonpolar amino acids refer to amino acids with hydrophobic side chain groups, including but not limited to alanine (Ala, A), valine (Val, V), leucine (Leu, L), isoleucine (Ile, I), phenylalanine (Phe, F), tryptophan (Trp, W), methionine (Met, M), and proline (Pro, P).

[0025] Basic amino acids are those whose side chains carry a positive charge at physiological pH. They specifically include arginine (Arg, R), lysine (Lys, K), and histidine (His, H).

[0026] Conservative substitution refers to the replacement of an original amino acid residue with another amino acid that has similar physicochemical properties (such as charge, size, and hydrophobicity), and usually does not significantly change the spatial conformation or function of the polypeptide.

[0027] This invention uses the wild-type sequence SEQ ID NO:2 as the leader sequence and designs a series of highly active variants based on the structure-activity relationship.

[0028] First, construct high-quality Mars scorpion (such as...) Figure 1 The venom gland tissue cDNA library (shown) was constructed by isolating and purifying total RNA and mRNA from scorpion venom glands, synthesizing the first and second strands of cDNA, ligating and transforming the double-stranded cDNA with the pSPORT1 vector, and obtaining the scorpion venom gland tissue cDNA library. Based on the constructed library, 10,000 clones were randomly selected for sequencing. Sequence analysis revealed that clone 286 was a novel antimicrobial peptide gene from Scorpion scorpion, named SmarAMP286, and its nucleotide sequence is shown in SEQ ID NO: 1.

[0029] The precursor organization of SmarAMP286 encodes 71 amino acid residues and consists of three parts: a signal peptide (22 residues), a mature peptide (14 residues), and a precursor peptide (35 residues). Figure 2 As shown, the inferred amino acid sequence is shown below the cDNA sequence; signal peptide amino acids are marked with a single underline; mature peptide amino acids are shown in purple shading; and amino acids in italics are C-terminal precursor peptides. Therefore, this invention provides a *Scorpionus martensii* polypeptide: KILARLWRAVRSIF (SEQ ID NO: 2).

[0030] On one hand, this invention also provides a group of point-mutated homologous peptides of the Smaramp286 polypeptide. Based on the mature peptide sequence (KILARLWRAVRSIF) of Smaramp286, its secondary structure was predicted using an online NPS@server [DSC (Discrimination of Protein Secondary Structure Class)], and its secondary structure image was displayed using the software AHTHEPROT 2000. The results show that Smaramp286 contains an α-Helix structure with a large number of net positively charged basic residues (Lys and Arg), making it a typical amphiphilic molecule. Based on the helix diagram of the polypeptide sequence, numerous amino acid point mutations were then performed on the KILARLWRAVRSIF of the Smaramp286 polypeptide sequence. The variant sequence has the structure of general formula I, X1ILX2X3LWX4X5VX6SIF, where X1, X2, X3, X4, X5, and X6 are amino acid substitution sites, corresponding to positions 1, 4, 5, 8, 9, and 11 of SEQ ID NO:2. The substituted amino acids are basic, nonpolar, or polar amino acids. Further, X1, X3, X4, and X6 were found to be selected from arginine, lysine, or histidine; X2 and X5 were selected from valine, glycine, or threonine. These variants do not affect its amphiphilic characteristics. The preferred sequences are SEQ ID NO:3 to SEQ ID NO:18.

[0031] On the other hand, this invention also provides a group of amino acid-deleted homologous peptides of the SmarAMP286 polypeptide. Based on the mature peptide sequence (KILARLWRAVRSIF) of SmarAMP286, its secondary structure was predicted using an online NPS@server [DSC (Discrimination of Protein Secondary Structure Class)], and its secondary structure image was displayed using the software AHTHEPROT 2000. The results showed that SmarAMP286 contains an α-Helix structure, exhibiting typical amphiphilic characteristics, and contains a large number of net positively charged basic residues (Lys and Arg). Based on the helix diagram of the polypeptide sequence, a large number of amino acid deletion mutations were then performed on the KILARLWRAVRSIF of the SmarAMP286 polypeptide sequence. The variant sequences are sequences formed by deleting 1 to 5 amino acids from general formula I, possessing the structure of general formula II, X1Y2Y3X2X3LWX4X5VX6Y 12 Y 13 Y 14The deleted amino acids correspond to the sites of the original nonpolar amino acids in SEQ ID NO:2. Specifically, X1, X3, X4, and X6 are sites where only amino acid substitution occurs, corresponding to SEQ ID NO:2, and are selected from basic amino acids or nonpolar amino acids, respectively; X2, X5, and Y... 12 The sites of amino acid substitution or deletion, corresponding to SEQ ID NO:2, are selected from basic amino acids, nonpolar amino acids, or are absent; Y2, Y3, Y 13 Y 14 For sites where only amino acid deletion occurs, Y2 is selected from isoleucine or is absent, Y3 is selected from leucine or is absent, Y... 13 Selected from leucine or absent, Y 14 Phenylalanine is selected or is absent; and the total number of missing amino acids in Formula II does not exceed 5. Preferred sequences are SEQ ID NO: 19 to SEQ ID NO: 45.

[0032] As described above, the peptides of the present invention can also be modified using chemical modification techniques known in the art. Modifications to the peptides of the present invention can be introduced during or at the end of peptide synthesis. For example, when synthesizing peptides using solid-phase synthesis techniques, N-terminal acetylation can be performed by reacting the amino acid sequence still bound to the resin with acetic acid. As another example, C-terminal amidation is performed in solid-phase peptide synthesis using specific types of resins containing chemical handles from which the amidated (poly)peptide is released during cleavage. These and other methods of modifying peptides are known to any person skilled in the art.

[0033] Salts of the polypeptides of the present invention are also provided. Such salts include, but are not limited to, acid addition salts and base addition salts. As used herein, a “pharmaceutically acceptable salt” of a polypeptide means a salt that retains the desired antimicrobial activity of the polypeptide and is suitable for administration to humans or animals. Methods for preparing salts of polypeptides are known in the art and generally involve mixing the polypeptide with a pharmaceutically acceptable acid or base, for example, by mixing the free acid or free base form of the product with one or more equivalents of a suitable acid or base in a solvent or medium in which the salt is insoluble or in a solvent (such as water, which is then removed by vacuum or lyophilization), or by exchanging the cation of an existing salt for another cation on a suitable ion exchange resin. Examples of pharmaceutically acceptable acids and bases include organic and inorganic acids such as formic acid, acetic acid, propionic acid, lactic acid, glycolic acid, oxalic acid, pyruvic acid, succinic acid, maleic acid, malonic acid, trifluoroacetic acid, cinnamic acid, sulfuric acid, hydrochloric acid, hydrobromic acid, nitric acid, perchloric acid, phosphoric acid, and thiocyanate, which form ammonium salts with the free amino group of polypeptides, and bases such as ethylamine, methylamine, dimethylamine, triethylamine, isopropylamine, diisopropylamine, and other monoalkyl, dialkyl, and trialkylamines, and ethylarylamine.

[0034] The peptides of the present invention can be prepared by various methods. For example, the peptides can be synthesized by commonly used solid-phase synthesis methods well known in the art, such as methods involving t-BOC or FMOC protection of the α-amino group. Here, an amino acid is subsequently added to the elongated amino acid chain. Such methods are described, for example, in "Solid Phase Peptide Synthesis," IRL Publishing, London. Solid-phase peptide synthesis methods are particularly suitable for the large-scale production of shorter peptides.

[0035] Alternatively, recombinant techniques well known in the art can be used to prepare the polypeptides of the present invention, wherein the nucleotide sequence encoding the polypeptide is expressed in a host cell. The present invention therefore provides a method for preparing the polypeptides of the present invention, comprising: Provide a nucleic acid molecule comprising a nucleic acid sequence encoding a polypeptide of the present invention; transform a host cell with the nucleic acid molecule; culture the host cell under conditions that allow expression of the polypeptide; harvest the polypeptide from the cell; optionally modify the polypeptide at its N-terminus or C-terminus, for example by adding an N-terminal and / or C-terminal extension group.

[0036] This invention also provides nucleic acid molecules comprising a nucleic acid sequence encoding the polypeptide of this invention, which are also referred to herein as nucleic acid molecules of this invention. The nucleic acid molecules or nucleic acid sequences of this invention, as used herein, comprise nucleotides, preferably DNA or RNA.

[0037] Another aspect of the invention is a pharmaceutical composition comprising at least one of the *Scorpionus martensii* polypeptides described herein. The composition may also comprise a pharmaceutically or pharmacologically acceptable carrier. The phrase "pharmaceutically or pharmacologically acceptable" means a molecular entity or composition that, when properly administered to an animal or human, will not produce adverse, allergic, or other adverse reactions. The compositions of the invention may be aqueous compositions comprising an effective amount of the *Scorpionus martensii* polypeptide dissolved or dispersed in a pharmaceutically acceptable carrier or aqueous medium.

[0038] The term "pharmaceutically acceptable carrier" as used in this invention includes any and all solvents, dispersion media, coatings, antimicrobial and antifungal agents, isotonic agents, and absorption delay agents, etc. The use of such media and reagents for pharmaceutically active substances is well known in the art. Their use in therapeutic compositions is also considered, except that any conventional media or reagent may be incompatible with the active ingredient. Additional active ingredients may also be incorporated into the composition.

[0039] The invention has now been generally described, and will be more readily understood by referring to the following embodiments, which are provided by way of example and not by way of limitation.

[0040] The strains used in this invention to test antibacterial activity are: E. coli AB94012 E. coli AB25922, P. aeruginosa AB93066 P. aeruginosa ATCC9027 A.baumannii ATCC19606 P. acnes ATCC6919 P. acnes ATCC11827 S. aureus AB94004 S. aureus ATCC6538 S. aureus ATCC25923, MRSA P1381, and MRSA P1474 are all bacteria well-known and familiar to those skilled in the art.

[0041] Example 1 The preparation of a polypeptide gene from *Scorpionus martensii* involves the following steps: A: Extraction of total RNA from the venom glands of Scorpion martensii (Trizol LS one-step method: Trizol LS purchased from Invitrogen, USA) ① Collect Mars's scorpion ( Figure 1 Take 500mg of scorpion tail gland ( Figure 1 ① Grind the scorpion venom gland into a fine powder in liquid nitrogen, add 20 ml of TRIZOL reagent, mix well, and incubate at room temperature (22-28℃, the same applies below) for 10 minutes; ② Then add 4 ml of chloroform, mix for 30 seconds, incubate at room temperature for 5 minutes, and centrifuge at 12000 g for 10 minutes at 4℃; ③ Take the aqueous phase, add 1 volume of isopropanol, incubate at room temperature for 20 minutes, and centrifuge at 12000 g for 20 minutes at 4℃ to obtain RNA precipitate; ④ Wash the precipitate with 10 ml of 75% ethanol and centrifuge at 10000 g for 5 minutes; ⑤ After drying the RNA precipitate, dissolve it in DEPC-treated water and incubate at 50-60℃ for 20 minutes to completely dissolve the RNA. The entire process was performed according to the recommended method of the TRlZOL (Total RNA Isolation) Reagent Kit. The quality of the prepared scorpion venom gland total RNA was detected by formaldehyde denaturing gel electrophoresis. High-quality scorpion venom gland total RNA was obtained.

[0042] B: mRNA isolation and purification mRNA was isolated and purified using the PolyA Tract mRNA isolation system (Promega, USA). Its working principle is based on the complementary pairing property of Oligo (dT) and the 3' poly(A) tail of mRNA. Biotin-labeled Oligo (dT) forms a hybrid through annealing with the 3' poly(A) tail of mRNA. The biotin-oligo (dT) / mRNA hybrid is then captured and washed using avidin-labeled magnetic beads and a magnetic separator. Finally, it is eluted with RNase-free sterile double-distilled water (ddH2O) to achieve the separation of mRNA from total RNA. ① Sample preparation: RNA was added to 800 μl of binding buffer containing 32 μl of β-mercaptoethanol. ② Probe annealing: Take 5 μl of 250 pM Oligo(dT) and add distilled water to a final volume of 50 μl; add 1.6 ml of preheated dilution buffer (with 32 μl of β-mercaptoethanol added), mix well with RNA, and incubate at 70°C for 5 minutes. ③ Activation of magnetic beads: Take 1.2 ml of SA-PMPS magnetic beads (purchased from Promega, USA) into a 1.5 ml centrifuge tube; resuspend SA-PMPS with 0.5×SSC, adsorb the magnetic beads onto a magnetic rack, and wash SA-PMPS 3 times with the original volume of 0.5×SSC. ④ mRNA Acquisition: Mix the RNA incubated at 70℃ with SA-PMPS, incubate at room temperature for 10 minutes, then place on a magnetic rack to adsorb magnetic beads and discard the supernatant; suspend the magnetic beads in 2 ml of 0.5X SSC, wash 4 times, removing as much SSC as possible in the last wash; add RNase-free ddH2O to the magnetic beads, mix gently, then centrifuge (12000 g × 3 minutes) or adsorb the magnetic beads on a magnetic rack; collect the supernatant to obtain mRNA. Determine the concentration and purity of mRNA by electrophoresis and UV. ⑤ mRNA Precipitation: Add glycogen and 2-3 volumes of anhydrous ethanol to the mRNA obtained in ④, precipitate overnight, and the mRNA will be used for cDNA synthesis.

[0043] C: First-strand cDNA synthesis ① Add 2 μl to a 1.5 ml Eppendorf tube Not ① Add the Primer-adapter and 6 μl mRNA (containing 3 μg mRNA), incubate at 70°C for 10 min, immediately place on ice, centrifuge, and then add the following components: 4 μl 5X first strand buffer; 2 μl 0.1M DTT; 1 μl 10mM dNTPs; 1 μl H2O. Gently mix and centrifuge, incubate at 37°C for 2 min; ② Add 5 μl reverse transcriptase, mix well, and take 2 μl, add 1 μl [α- 32P]dCTP (4 μCi) (tracer tube). Incubate simultaneously with the above reaction components (sample tube) at 37°C for 1 h, then place on ice to terminate the reaction; ③ For the tracer tube, add 43 μl of 20 mM EDTA and 5 μl of yeast tRNA sequentially, mix well, and then take two 10 μl portions and spot them onto two filter membranes. Wash one portion three times with 10% TCA for 5 min each time, wash once with 95% ethanol, air dry, and then place in 1.5 ml of scintillation solution (for 1 # One sample); another sample, after air-drying, was placed in 1.5 ml of scintillation solution (for 2 samples). # (Sample). Add another 30 µl of tracer solution, 1.5 μl of 7.5 M ammonium acetate (NH4OAc), and 90 μl of anhydrous ethanol (-20℃). Mix well and immediately centrifuge at 14,000 rpm for 20 min. Discard the supernatant, add 0.5 ml of 70% anhydrous ethanol (-20℃), centrifuge at 14,000 rpm for 2 min, discard the supernatant, and dry at 37℃ for 10 min to allow the ethanol to evaporate. Dissolve in 10 μl of TEN solution, add 10 μl of 2X loading buffer, and use 10 μl for alkaline gel electrophoresis. Use [a- 32 P]dCTP-labeled λDNA Hind ④ After mixing, incubate at room temperature for 15 min, then add 2 μl of 0.2 M EDTA to terminate the reaction. Take 6 μl of the reaction solution and 6 ml of 2X alkaline electrophoresis buffer, mix well, electrophore for 5 h, then soak in 7% TCA for 20 min until bromophenol blue turns yellow. Then blot dry with tissue paper (about 10 h) and perform autoradiography; ⑤ For the sample tube, use it for the synthesis of the second chain.

[0044] Hind III 10X buffer 2 μl dGTP0.2 mM dATP 0.2 mM [α- 32 P]dCTP2 μCi Hind III markers 1 μg Klenow DNA polymerase2 unit Add ddH2O to a total volume of 20 μl D: Second-strand cDNA synthesis ① Add the following components sequentially to the sample tube on ice; ② Gently mix and incubate at 16°C for 2 h; ③ Add 2 μl (10 units) of T4 DNA polymerase and continue the reaction at 16°C for 5 min; ④ Transfer to ice and add 10 μl of 0.5 M EDTA; ⑤ Add an equal volume (150 μl) of phenol / chloroform / isoamyl alcohol (25 / 24 / 1), vortex thoroughly, and centrifuge at 14,000 rpm for 5 min at room temperature. Transfer the aqueous phase (140 μl) to another 1.5 ml Eppendorf tube; ⑥ Add 70 μl of 7.5 M NH4OAc and 0.5 ml of anhydrous ethanol (-20°C), vortex, and centrifuge at 14,000 rpm for 20 min at room temperature; ⑦ Discard the supernatant, add 0.5 ml of 70% ethanol (-20°C), and centrifuge for 2 min as above. Discard the supernatant and dry at 37°C for 10 min.

[0045] DEPC-treated water 92 μl 5X second strand buffer 30 μl 10 mM dNTP mix 3 μl E.coli DNA ligase (10units / μl)1 μl E.coli DNA polymerase (10units / μl)4 μl E.coli RNase H (2units / μl)1 μl Total volume 150 μl E: Double-stranded cDNA and Sal I connector connection ① Dissolve the cDNA sample of sample D in 25 μl of sterile water, then add the following ingredients in the order listed in the table below; ② Mix gently and react at 16°C overnight (approximately 20 h); ③ Extract with phenol / chloroform / isoamyl alcohol (25 / 24 / 1) and precipitate with NH4OAc / ethanol, then dry at 37°C for 10 min.

[0046] 5X T4 DNA ligase buffer 10 μl Sal I adapters 10 μl T4 DNA ligase 5 μl Total volume 50 μl F: Not I digest double-stranded cDNA ① Dissolve sample E in 41 μl, then add the following ingredients sequentially according to the table below; ② Mix well and incubate at 37℃ for 2 h; ③ Extract once with phenol / chloroform / isoamyl alcohol (25 / 24 / 1), then precipitate with 7.5 M NH4OAc / ethanol and dry at 37℃ for 10 min; ④ Dissolve in 70 μl TEN, take 1 μl for quantification, and store the remainder at -20℃ for later use.

[0047] REACT 3 buffer 5 μl Not I4 μl Total volume 50 μl G: Remove excess cDNA molecules Sal I. Connector and enzyme-digested fragments Remove excess using the nucleon extraction and purification kit (Amersham, USA). Sal I. Connector and Enzyme Digestion Fragment. ① Suspend the resin at room temperature, then add 600 μl to a centrifuge column, centrifuge at 2000 rpm for 10 s, and discard the liquid. Add 40 μl of the above cDNA solution to the center of the resin. Centrifuge as above; ② Collect the eluent for the ligation reaction.

[0048] H: Ligation and transformation of double-stranded cDNA with pSPORT1 vector ① Add the following components sequentially to a 1.5 ml Eppendorf tube; ② React at room temperature for 16 h; ③ Add the following components sequentially to the reaction solution in ②: 5.0 μl yeast tRNA, 12.5 μl 7.5 M NH4OAc, 70 μl anhydrous ethanol (-20℃). Vortex to mix and immediately centrifuge at 14000 rpm for 20 min; ④ Wash the precipitate with 70% ethanol (-20℃), dry at 37℃, and dissolve in 4 μl; ⑤ Electrolyze 2 μl of the precipitate into 50 μl of the precipitate. E. coli K12 MC1061. The quality of the library was determined by PCR. Forward primer: 5'-TCGACCCACGCGTCCG-3' (designed according to the SalI adapter sequence); Reverse primer: 5'-GAGCGGCCGCCCT15-3' (designed according to the NotI primer-adaptor sequence).

[0049] 5X T4 DNA ligase buffer 4 μl pSPORT1, Not I-Sal I-Cut (50ng / μl)1 μl cDNA (3ng / μl) 4 μl T4 DNA ligase 1 μl Add ddH2O to a total volume of 20 μl I: Random sequencing strategy for screening cDNA libraries 10,000 clones were randomly selected from the constructed cDNA library of *Scorpion musculus* venom glands and sent to Shanghai Sanbo Biotechnology Co., Ltd. for sequencing. The sequence entry software was BioEdit v4.5.8 (Tom Hall, 1999), and the homology comparison and signal peptide cleavage site prediction software were CLUSTAL X 1.8 (Thompson et al., 1997) and PC / GENE (Intelligenetics Inc., Switzerland), respectively. Sequence analysis showed that clone 286 was a novel antimicrobial peptide gene, named SmarAMP286, with its nucleotide sequence shown in SEQ ID NO: 1. The precursor organization form of SmarAMP286 encodes 71 amino acid residues, consisting of three parts: a signal peptide (22 residues), a mature peptide (14 residues), and a precursor peptide (35 residues). Figure 2 Therefore, the present invention provides a Mars scorpion polypeptide: KILARLWRAVRSIF (SEQ ID NO: 2).

[0050] Example 2 Structural analysis of the SmarAMP286 peptide and its point-mutated homologous amphiphilic peptide.

[0051] Based on the mature peptide sequence (KILARLWRAVRSIF) of SmarAMP286, its secondary structure was predicted using the online NPS@server [DSC (Discrimination of Protein Secondary Structure Class)], and its secondary structure image was displayed using the AHTHEPROT 2000 software. The results showed that SmarAMP286 contains an α-Helix structure with a large number of net positively charged basic residues (Arg and Lys), making it a typical amphiphilic molecule. Based on the helix diagram of the peptide sequence, numerous amino acid point mutations were performed on the KILARLWRAVRSIF sequence of SmarAMP286. It was found that the sequence X1ILX2X3LWX4X5VX6SIF (X1, X3, X4, and X6 are Lys, Arg, or His; X2 and X5 are Ala, Gly, or Thr) did not affect its amphiphilic characteristics. Therefore, the present invention provides a group of point-mutated structural homologous peptides of SmarAMP286 polypeptide (Table 1, SEQ ID NO: 3-18).

[0052] Example 3 SmarAMP286 peptide is an amino acid-deficient homologous amphiphilic peptide.

[0053] Based on the mature peptide sequence of SmarAMP286 (KILARLWRAVRSIF), its secondary structure was predicted using the online NPS@server [DSC method (Discrimination of protein Secondary structure Class)]. The secondary structure images of homologous peptides with amino acid deletions were then displayed using AHTHEPROT 2000 software. Variant sequences are formed by deleting 1 to 5 amino acids from general formula I, exhibiting the structure of general formula II: X1Y2Y3X2X3LWX4X5VX6Y 12 Y 13 Y 14 The deleted amino acids correspond to the sites of the original nonpolar amino acids in SEQ ID NO:2. Specifically, X1, X3, X4, and X6 are sites where only amino acid substitution occurs, corresponding to SEQ ID NO:2, and are selected from basic amino acids or nonpolar amino acids, respectively; X2, X5, and Y... 12 The sites of amino acid substitution or deletion, corresponding to SEQ ID NO:2, are selected from basic amino acids, nonpolar amino acids, or are absent; Y2, Y3, Y 13 Y 14 For sites where only amino acid deletion occurs, Y2 is selected from isoleucine or is absent, Y3 is selected from leucine or is absent, Y... 13 Selected from leucine or absent, Y 14 Phenylalanine is selected or is absent; and the total number of missing amino acids in Formula II does not exceed 5.

[0054] Based on the helix diagram of the polypeptide sequence, and then by performing a large number of amino acid deletion mutations on the SmarAMP286 polypeptide sequence KILARLWRAVRSIF, it was found that X7X8LWX9X 10 VX 11 SIF(X7, X8, X9 and X 11 For Lys, Arg, or His; X 10 The sequences Ala, Gly, or Val (SEQ ID NO:4) and HKLWKFVKR do not affect their amphipathic characteristics. Therefore, this invention provides a group of amino acid-deficient homologous peptides of the SmarAMP286 polypeptide (Table 2, SEQ ID NO:19 to SEQ ID NO:45).

[0055] Example 4 Chemical synthesis of SmarAMP286 polypeptide and its structurally homologous amphiphilic polypeptide Based on the amino acid sequences of SmarAMP286 and its structurally homologous amphiphilic polypeptides (KILARLWRAVRSIF, X1ILX2X3LWX4X5VX6SIF, X7X8LWX9X...),... 10 VX 11 SIF and HKLWKFVKR were used for artificial synthesis. Solid-phase chemical synthesis yielded high-purity (95% purity) SmarAMP286 peptides with the correct molecular weight. Figure 3 and Figure 4 ) and its structurally homologous amphiphilic polypeptides (Table 1 and Table 2).

[0056]

[0057]

[0058] Example 5 Minimum inhibitory concentrations (MICs) of SmarAMP286 polypeptide and its structurally homologous amphiphilic polypeptide against Escherichia coli, Pseudomonas aeruginosa, Acinetobacter baumannii, Propionibacterium acnes, Staphylococcus aureus, and related drug-resistant bacteria.

[0059] The application process of SmarAMP286 polypeptide and its structurally homologous amphiphilic polypeptide in the preparation of drugs for the treatment or prevention of infections caused by Escherichia coli, Pseudomonas aeruginosa, Acinetobacter baumannii, Propionibacterium acnes, Staphylococcus aureus, and related drug-resistant bacteria is as follows: A- Escherichia coli, Pseudomonas aeruginosa, Acinetobacter baumannii, Propionibacterium acnes, Staphylococcus aureus, and related drug-resistant bacteria are cultured separately at OD... 630 When the concentration is 0.8, after diluting 400 times, take 80 µl and add it to a 96-well plate. Then, add 20 µl of the serially diluted SmarAMP286 peptide or its structural homologous amphiphilic peptide to each well to achieve a final concentration of 200 µg / ml, 100 µg / ml, 50 µg / ml, 25 µg / ml, 12.5 µg / ml, 6.25 µg / ml, 3.125 µg / ml or 160 µg / ml, 80 µg / ml, 40 µg / ml, 20 µg / ml, 10 µg / ml, 5.0 µg / ml, 2.5 µg / ml. Add 20 µl of 1% BSA to the negative control wells; after incubation at 37℃ for 12 hours, measure the absorbance of each well in the 96-well plate at 630 nM using a microplate reader; after determining the minimum inhibitory concentration (MIC) of the drug SmarAMP286 peptide and its structural homologous amphiphilic peptide at a 2-fold dilution, repeat the experiment three times. Finally, determine the MIC of SmarAMP286 peptide and its structural homologous amphiphilic peptide against *Escherichia coli*, *Pseudomonas aeruginosa*, *Acinetobacter baumannii*, *Propionibacterium acnes*, *Staphylococcus aureus*, and related drug-resistant bacteria.

[0060] The antibacterial results showed that SmarAMP286 peptide and its structurally homologous amphiphilic peptides had good antibacterial activity and efficacy against Escherichia coli, Pseudomonas aeruginosa, Acinetobacter baumannii, Propionibacterium acnes, Staphylococcus aureus and related drug-resistant bacteria, with a minimum inhibitory concentration (MIC) range of 5 µg / ml to 100 µg / ml (as shown in Tables 3 and 4).

[0061] Table 3: Minimum inhibitory concentrations of SmarAMP286 polypeptide from Mars and its point-mutated structural homologs in different bacteria.

[0062]

[0063] Table 4: Minimum inhibitory concentrations of SmarAMP286 polypeptide from Mars and its amino acid-deficient homologous polypeptides in different bacteria.

[0064]

[0065] Example 6 Preparation of antibacterial agents of SmarAMP286 polypeptide and its structurally homologous amphiphilic polypeptide.

[0066] (1) Prescription: 0.1 g of SmarAMP286 polypeptide and its structural homologous amphiphilic polypeptide, 10 g of propylene glycol, 20 g of ethanol, 1.5 g of hydroxymethylcellulose, 1 g of triethanolamine, 0.1 g of ethylparaben, and sterile water to 100 g.

[0067] (2) Preparation: Sprinkle hydroxymethyl cellulose on the liquid surface (about 60 ml), and after overnight storage, it becomes a gel matrix. After standing, degas under vacuum. Mix triethanolamine with the remaining drugs and gradually add it to the slurry and mix well, avoiding vigorous stirring to prevent the introduction of too many air bubbles. Dispense into the product to obtain the antibacterial gel preparation.

[0068] (3) Blank control gel: Except that it does not contain SmarAMP286 polypeptide and its structurally homologous amphiphilic polypeptide, the formulation and preparation are the same as the above formulations.

[0069] Example 7 SmarAMP286 and its structurally homologous amphiphilic polypeptide antibacterial agents inhibit Escherichia coli, Pseudomonas aeruginosa, Acinetobacter baumannii, Propionibacterium acnes, Staphylococcus aureus, and related drug-resistant bacteria.

[0070] (1) Dissolve the finished gel in sterile saline at a ratio of 1:2 and dilute it into 6 equal concentrations, and place it in a refrigerator at 4°C.

[0071] (2) The preserved Escherichia coli, Pseudomonas aeruginosa, Acinetobacter baumannii, Propionibacterium acnes, Staphylococcus aureus and related drug-resistant bacteria were inoculated into LB medium and cultured overnight at 37°C.

[0072] (3) Dilute the overnight cultured bacterial solution with fresh LB medium to OD. 630 =0.002.

[0073] (4) Add 80 µl of the above bacterial solution to each well of the 96-well culture plate.

[0074] (5) Add 20 µl of the above-mentioned finished gel solutions of different concentrations to the bacterial solution.

[0075] (6) Simultaneously perform negative control tests for gel and saline, as well as positive control tests for raw material peptides and antibiotics.

[0076] (7) Place the 96-well plate on a shaker at 37°C and 250 rpm for 16 hours.

[0077] (8) After 16 hours, remove the 96-well plate, cool it to room temperature, and place it on an ELISA reader to measure its absorbance at a wavelength of 630 nm.

[0078] (9) The minimum inhibition concentration is the concentration of the solution with no light absorption at all.

[0079] (10) The antibacterial results showed that the antibacterial preparation had a good inhibitory effect on Escherichia coli, Pseudomonas aeruginosa, Acinetobacter baumannii, Propionibacterium acnes, Staphylococcus aureus and related drug-resistant bacteria. Its minimum inhibitory concentration ranged from 5 µg / ml to 100 µg / ml (calculated for the active ingredient peptide in the preparation), which was consistent with the minimum inhibitory concentration of the active pharmaceutical ingredient peptide (the MIC of the active pharmaceutical ingredient SmarAMP286 and its structurally homologous amphiphilic synthetic peptide against Escherichia coli, Pseudomonas aeruginosa, Acinetobacter baumannii, Propionibacterium acnes, Staphylococcus aureus and related drug-resistant bacteria was between 5 µg / ml and 100 µg / ml), while the blank preparation had no antibacterial activity (Tables 5 and 6).

[0080] Table 5: Minimum inhibitory concentrations (MICs) of SmarAMP286, a polypeptide from Mars and its point-mutated homologous polypeptides, against Escherichia coli, Pseudomonas aeruginosa, Acinetobacter baumannii, Propionibacterium acnes, Staphylococcus aureus, and related drug-resistant bacteria in topical gel formulations (calculated based on polypeptide content in the formulation).

[0081]

[0082] Table 6: Minimum inhibitory concentrations (MICs) of SmarAMP286 and its amino acid-deficient homologous peptides in topical gel formulations against Escherichia coli, Pseudomonas aeruginosa, Acinetobacter baumannii, Propionibacterium acnes, Staphylococcus aureus, and related drug-resistant bacteria (calculated based on peptide content in the formulation).

[0083]

[0084] Example 8 Clinical efficacy of SmarAMP286 and its structurally homologous amphiphilic peptide antibacterial agents in acne patients.

[0085] (1) Case selection: All patients included were clinically diagnosed with acne and came from the dermatology outpatient clinic of Renmin Hospital of Wuhan University. Patients with a history of allergy to clindamycin or quinolone, those who had used other anti-acne drugs within 15 days, those with severe liver or kidney dysfunction, and pregnant or lactating women were excluded.

[0086] (2) Experimental grouping: 368 patients were divided into an experimental group and a control group using a multicenter open-label parallel controlled observation method. Experimental group: 228 patients, 106 males and 122 females, aged 18 to 45 years. Control group: 140 patients, 72 males and 68 females, aged 18 to 46 years. There were no significant differences in age, gender, disease stage, and skin lesion severity between the two groups, making them comparable.

[0087] (3) Experimental methods: The experimental group was given the topical medication for treating acne, namely SmarAMP286 and its structurally homologous amphiphilic polypeptide topical gel of the present invention; the control group was given 5% sulfur cream (made by the preparation room of Wuhan University People's Hospital) for topical application. Method of application: Wash the face with warm water and soap or sulfur soap to thoroughly clean the facial oil and dirt, then use your fingers to apply the medicine repeatedly and gently to the affected area, once in the morning and once in the evening, for two consecutive weeks as one course of treatment.

[0088] (4) Observation and judgment criteria for efficacy: 368 participants were followed up once a week. During the follow-up visit, the observation form was filled out in detail, recording the number, changes and adverse reactions of facial acne lesions. The efficacy was calculated based on the total percentage reduction of various acne lesions (acne, inflammatory scars, pustules, nodules, cysts). Cure: 100% lesion reduction; Significant effect: 76-99% lesion reduction; Effective: 50-75% lesion reduction; Ineffective: <50% lesion reduction. After the course of treatment, the percentage of acne after treatment was calculated using a quantitative method to evaluate the efficacy. At the same time, adverse reactions were observed, including local irritation and systemic symptoms. Some patients underwent routine blood and urine tests and liver and kidney function tests before and after the clinical trial.

[0089] (5) Statistical analysis of efficacy: In the experimental group of 228 cases, 142 cases were cured, accounting for 62.3%; 66 cases showed significant improvement, accounting for 28.9%; 5 cases were effective, accounting for 8.8%; and 0 cases were ineffective, accounting for 0%; the total effective rate was 100%. In the control group of 140 cases, 7 cases were cured, accounting for 5.0%; 16 cases showed significant improvement, accounting for 11.4%; 45 cases were effective, accounting for 32.1%; and 72 cases were ineffective, accounting for 51.4%; the total effective rate was 48.5%. The efficacy analysis showed a highly significant difference between the experimental group and the control group, indicating that the experimental group was more effective than the control group in treating acne.

[0090] (6) Adverse reactions: No adverse reactions were observed in any of the 228 patients in the experimental group. In the control group, 9 out of 140 patients experienced local burning and redness after applying the medication, which subsided spontaneously after discontinuation of the medication without further treatment. In addition, routine blood and urine tests were performed in 20 patients, liver function tests were performed in 30 patients, and kidney function tests were performed in 15 patients before and after treatment in the experimental group, and no abnormalities were found in any of them.

[0091] Patients with acne, pimples, and blackheads treated at the hospital were selected as subjects. All patients were randomly divided into an experimental group and a control group. The experimental group received a topical antibacterial gel containing scorpion polypeptide, while the control group received routine medication (5% sulfur cream). The number, changes, and adverse reactions of facial acne lesions were recorded in both groups. The efficacy was calculated based on the total percentage reduction in various acne lesions (blackheads, inflammatory scars, pustules, nodules, cysts). The effective rate in the experimental group using the topical antibacterial gel containing scorpion polypeptide reached 100%, while the effective rate in the control group using routine medication was 48.5%. The difference in efficacy between the two groups was highly significant, indicating that the experimental group was more effective than the control group in treating acne.

[0092] This invention provides a highly effective topical gel preparation for treating acne. Patients with acne, pimples, and blackheads treated at a hospital were selected as subjects, and all patients were randomly divided into an experimental group and a control group. The experimental group used a scorpion polypeptide topical gel antibacterial preparation, while the control group received conventional medication (5% sulfur cream). The number, changes, and adverse reactions of facial acne lesions were recorded in both groups. The efficacy was statistically analyzed based on the total percentage reduction in various acne lesions (blackheads, inflammatory scars, pustules, nodules, cysts). The effective rate of the scorpion polypeptide topical gel antibacterial preparation in the experimental group reached 100%, while the effective rate in the control group receiving conventional medication was 48.5%. The difference in efficacy between the two groups was highly significant, indicating that the experimental group was more effective than the control group in treating acne.

[0093] This invention is a novel and more effective acne treatment formulated with active polypeptides from the traditional Chinese medicinal herb scorpion as the active ingredient. It demonstrates significant efficacy and remarkable therapeutic effects in treating acne. The efficacy is excellent; for patients with mild acne, pimples, or zits, symptoms disappear in just 2-3 days, and even severe cases in 5-12 days, with an effectiveness rate of 100% and a cure rate of approximately 62.3%, and no recurrence after treatment. The raw materials used in this invention are all active ingredients from the traditional Chinese medicinal herb scorpion, with no toxic side effects and no sequelae. This medicine truly achieves rapid effectiveness, no side effects, and no recurrence after treatment.

[0094] Where there is no conflict, the above embodiments and features described herein can be combined with each other.

[0095] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. An isolated Eremobates polypeptide or a pharmaceutically acceptable salt thereof, characterized in that, The Myrmales scorpion polypeptide comprises a sequence of SEQ ID NO: 2 or a variant sequence thereof, the variant sequence comprising an amino acid sequence of general formula I or general formula II, The general formula I is X1ILX2X3LWX4X5VX6SIF, wherein X1, X2, X3, X4, X5, X6 are amino acid substitution sites corresponding to positions 1, 4, 5, 8, 9, 11 of SEQ ID NO: 2, and the amino acids for substitution are basic amino acids, non-polar amino acids or polar amino acids; The general formula II is a sequence formed by deletion of 1 to 5 amino acids from the general formula I, the deleted amino acids corresponding to the positions of the original non-polar amino acids in SEQ ID NO: 2; and the Myrmales scorpion polypeptide has an activity of inhibiting gram-negative bacteria and / or gram-positive bacteria.

2. The Maratha scorpion polypeptide of claim 1, wherein, The general formula II is formed by deletion of one or more amino acids selected from the following structures: X1Y2Y3X2X3LWX4X5VX6 Y 12 Y 13 Y 14 wherein, X1, X3, X4, X6 are only amino acid substitution sites, corresponding to SEQ ID NO: 2, and are selected from basic amino acids or non-polar amino acids, respectively; X2, X5, Y 12 For the sites where amino acid substitution or deletion occurs, corresponding to SEQ ID NO: 2, are selected from among basic amino acids, non-polar amino acids, or nonexistence, respectively; Y2, Y3, Y 13 , Y 14 is a site of only amino acid deletion, Y2 is selected from isoleucine or absent, Y3 is selected from leucine or absent, Y 13 is selected from leucine or absent, Y 14 is selected from phenylalanine or absent; and the total number of deleted amino acids in general formula II is not more than 5.

3. The Maratha scorpion polypeptide according to claim 1 or 2, characterized in that, The basic amino acid is selected from arginine, lysine or histidine; the non-polar amino acid is selected from alanine, valine, leucine, isoleucine, phenylalanine, tryptophan or methionine.

4. The Maratha scorpion polypeptide of claim 1, wherein, In the general formula I: X1, X3, X4 and X6 are selected from arginine, lysine or histidine; X2, X5 are selected from valine, glycine or threonine.

5. The Maratha scorpion polypeptide according to any one of claims 1 to 4, characterized in that, The amino acid sequence of the Myrmales scorpion polypeptide is selected from SEQ ID NO: 3 to SEQ ID NO:

45.

6. An isolated nucleic acid molecule, comprising, A nucleotide sequence encoding the Myrmales scorpion polypeptide according to any one of claims 1-5.

7. A pharmaceutical composition, characterized by, A pharmaceutical composition comprising an effective dose of the Myrmales scorpion polypeptide according to any one of claims 1-5 and a pharmaceutically acceptable carrier.

8. Use of the Myrmales scorpion polypeptide according to any one of claims 1-5 in the preparation of a medicament for treating or preventing bacterial infection diseases, the bacteria including Escherichia coli, Pseudomonas aeruginosa, Acinetobacter baumannii, Propionibacterium acnes, Staphylococcus aureus and their related drug-resistant bacteria.

9. Use according to claim 8, characterized in that, The medicament comprises a drug for treating or preventing acne, acne vulgaris, and comedones.

10. An external use gel preparation, characterized by, The Myrmales scorpion polypeptide according to any one of claims 1-5 and a gel base are formulated, and the gel base is hydroxymethyl cellulose.

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