CN114349825B - Scorpion venom derivative peptide and application thereof in preparation of antibacterial or anti-inflammatory drugs - Google Patents
Scorpion venom derivative peptide and application thereof in preparation of antibacterial or anti-inflammatory drugs Download PDFInfo
- Publication number
- CN114349825B CN114349825B CN202111680590.3A CN202111680590A CN114349825B CN 114349825 B CN114349825 B CN 114349825B CN 202111680590 A CN202111680590 A CN 202111680590A CN 114349825 B CN114349825 B CN 114349825B
- Authority
- CN
- China
- Prior art keywords
- scorpion venom
- derived peptide
- antibacterial
- peptide
- ffg
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Peptides Or Proteins (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention belongs to the technical field of polypeptide medicaments, and discloses a scorpion venom derivative peptide and application thereof in preparation of antibacterial or anti-inflammatory medicaments. The amino acid sequence of the scorpion venom derivative peptide is as follows: FFGRLFKLAVKIIPSLFK. The scorpion venom derived peptide has low toxicity to normal cells and good safety; the scorpion venom derived peptide has remarkable inhibition effect on bacteria such as escherichia coli, staphylococcus aureus, pseudomonas aeruginosa and the like; meanwhile, the expression of downstream inflammatory factors can be influenced by the regulation and control of NF- κB, so as to play the roles of anti-inflammation and immunoregulation.
Description
Technical Field
The invention belongs to the technical field of polypeptide medicaments, and particularly relates to a scorpion venom derivative peptide and application thereof in preparation of antibacterial or anti-inflammatory medicaments.
Background
Escherichia coli (E.coli) is a gram-negative short bacterium that can cause urinary tract, blood, skin and soft tissue infections, and serious infections can be life threatening. And because of numerous Escherichia coli serotypes, the immunoprophylaxis effect is not ideal, so that the antibacterial drug is still the main treatment scheme at present. However, due to the overuse of broad-spectrum antibiotics, the degree of resistance of Escherichia coli becomes more and more severe, and the resistant strain has the ability to accumulate resistance genes by horizontal gene transfer, resulting in a decrease in the efficacy of clinically existing antibiotics or even complete failure. Therefore, the development of novel antibacterial drugs which are not easy to cause bacteria to generate drug resistance is of great significance for clinical treatment.
Antibacterial peptides (AMPs) are a class of small peptide substances that are widely found in natural organisms, and constitute the first line of defense of hosts against infection by foreign pathogens. Antimicrobial peptides have three major advantages over traditional antibiotics: 1. the antibacterial peptide has a net positive charge, so that the antibacterial peptide can be preferentially combined with a cell membrane with negative charges of microorganisms, and the cell membrane is an inherent structure of organisms, so that the antibacterial peptide has a wide inhibition effect on bacteria, fungi, parasites, viruses, tumor cells and the like; 2. because of the specificity of the prokaryotic and eukaryotic cell membrane lipid, the antibacterial peptide has the characteristic of selective sterilization and has lower toxicity to normal eukaryotic cells; 3. the target molecule of the antibacterial peptide is a conserved cell membrane component, and drug resistance mechanisms such as enzymolysis, extracellular discharge drugs and the like are not existed in the antibacterial process of the antibacterial peptide, so that the drug resistance is extremely difficult to generate. The above advantages make the antibacterial peptide widely paid attention to, and the antibacterial peptide is hopeful to replace the existing antibiotics to be used for treating the infection of the clinical common drug-resistant strain.
Scorpion toxin is widely distributed as a biological toxin, can generate various pharmacological activities due to complex components and properties, and has development and application potential. In the early stage, the scorpion toxin monomer (the amino acid sequence of which is FLSSLIPSAISGLISAFK) with obvious inhibition effect on the growth of escherichia coli is obtained by separating and screening the scorpion from the yellow fat tail scorpion (Androctonus australis). After analysis of the sequence structure, we found that there was room for further improvement in antibacterial ability. Therefore, the invention designs and modifies the structure of the selected scorpion toxin monomer to obtain the synthetic scorpion toxin derivative peptide with better activity.
Disclosure of Invention
The present invention aims to solve at least one of the technical problems in the prior art described above. Therefore, the invention provides a scorpion venom derivative peptide and application thereof in preparing antibacterial or anti-inflammatory medicaments. The scorpion venom derived peptide has low toxicity to normal cells and good safety; the scorpion venom derived peptide has remarkable inhibition effect on bacteria such as escherichia coli, staphylococcus aureus, pseudomonas aeruginosa and the like; meanwhile, the expression of downstream inflammatory factors can be influenced by the regulation and control of NF- κB, so as to play the roles of anti-inflammation and immunoregulation.
The invention provides a scorpion venom derivative peptide, which has the amino acid sequence as follows: FFGRLFKLAVKIIPSLFK (SEQ ID NO: 1).
The scorpion venom derivative peptide is an antibacterial polypeptide independently developed by the applicant, is a novel scorpion venom derivative peptide which is obtained by structurally modifying a scorpion venom monomer (amino acid sequence FLSSLIPSAISGLISAFK) separated from a yellow fat tail scorpion and is screened out by a molecular design and amino acid scanning substitution method, and has the amino acid sequence as follows: FFGRLFKLAVKIIPSLFK (SEQ ID NO: 1), designated FFG-18. The scorpion venom derived peptide polypeptide contains 18 amino acid residues, the average molecular weight is 2124.65g/mol, the hydrophobicity coefficient is 1.03, and the theoretical isoelectric point pH is 11.73. The scorpion venom derivative peptide FFG-18 can be synthesized by adopting a solid-phase synthesis method, and has the characteristics of convenience and rapidness in synthesis, stability and controllability; the scorpion venom derived peptide FFG-18 can also be synthesized by a biosynthesis method.
Preferably, the amino acids in the scorpion venom derived peptide are all L-amino acids.
The invention also provides an isolated nucleic acid molecule which can code the scorpion venom derivative peptide.
The invention also provides a recombinant vector comprising the nucleic acid molecule.
The invention also provides application of the scorpion venom derivative peptide or pharmaceutically acceptable salt thereof in preparation of antibacterial products.
Experiments show that the scorpion venom derived peptide has remarkable antibacterial effect, has good inhibition and killing effects on various bacteria, and can be used as an antibacterial drug in disease treatment or used as an antibacterial component in disinfectants, cleaners, preservatives and packaging materials.
Preferably, the pharmaceutically acceptable salt of the scorpion venom derived peptide comprises at least one of acetate and citrate. In order to improve the stability or solubility of polypeptide drugs, the scorpion venom derived peptides can be prepared in the form of acetate or citrate.
The invention also provides an antibacterial drug which comprises the scorpion venom derivative peptide and/or pharmaceutically acceptable salt thereof and pharmaceutically acceptable auxiliary materials.
Preferably, the bacterial species that the antibacterial agent inhibits/kills include escherichia coli, staphylococcus aureus, and pseudomonas aeruginosa. Wherein, the antibacterial drug has more remarkable inhibiting/killing effect on escherichia coli.
Preferably, the pharmaceutically acceptable auxiliary materials are at least one of solvents, wetting agents, emulsifying agents, thickening agents, excipients, suspending agents, disintegrating agents, filling agents, lubricating agents or diluents.
Preferably, the dosage form of the antibacterial drug is tablets, injections, sprays, freeze-dried powder injections, capsules or coated pills.
More preferably, the dosage form of the antibacterial agent is an injection. As the scorpion venom derivative peptide belongs to polypeptide compounds, the scorpion venom derivative peptide is easy to degrade and difficult to pass through intestinal mucosa, and the drug effect of the scorpion venom derivative peptide can be reduced by oral administration, so that the scorpion venom derivative peptide is prepared into an injection form, and the scorpion venom derivative peptide is more beneficial to absorption and utilization of drug components.
The invention also provides application of the scorpion venom derivative peptide or pharmaceutically acceptable salt thereof in preparing anti-inflammatory medicaments.
Experiments show that the scorpion venom derived peptide can influence the expression of downstream inflammatory factors through the regulation and control of NF- κB, and has good anti-inflammatory and immunoregulatory effects.
Preferably, the pharmaceutically acceptable salt of the scorpion venom derived peptide comprises at least one of acetate and citrate.
The invention also provides an anti-inflammatory drug which comprises the scorpion venom derivative peptide and/or pharmaceutically acceptable salt thereof and pharmaceutically acceptable auxiliary materials.
Preferably, the pharmaceutically acceptable auxiliary materials are at least one of solvents, wetting agents, emulsifying agents, thickening agents, excipients, suspending agents, disintegrating agents, filling agents, lubricating agents or diluents.
Preferably, the anti-inflammatory medicament is in the form of a tablet, an injection, a spray, a freeze-dried powder injection, a capsule or a coated pill.
More preferably, the anti-inflammatory agent is in the form of an injection. As the scorpion venom derivative peptide belongs to polypeptide compounds, the scorpion venom derivative peptide is easy to degrade and difficult to pass through intestinal mucosa, and the drug effect of the scorpion venom derivative peptide can be reduced by oral administration, so that the scorpion venom derivative peptide is prepared into an injection form, and the scorpion venom derivative peptide is more beneficial to absorption and utilization of drug components.
Compared with the prior art, the invention has the following beneficial effects:
(1) The scorpion venom derived peptide (FFG-18) has low toxicity to normal cells and good safety;
(2) The scorpion venom derivative peptide (FFG-18) can directly destroy bacterial cell membranes through physical action, so that the sterilization speed is high; the scorpion venom derived peptide (FFG-18) has remarkable inhibition effects on escherichia coli, staphylococcus aureus, pseudomonas aeruginosa and the like, is not easy to generate drug resistance, and has remarkable inhibition effects on clinically isolated drug-resistant strains through tests.
(3) The scorpion venom derivative peptide (FFG-18) can influence the expression of downstream inflammatory factors through the regulation and control of NF- κB, and has anti-inflammatory and immunoregulatory effects.
Drawings
FIG. 1 shows the HPLC data for scorpion venom derived peptide FFG-18;
FIG. 2 shows a mass spectrum data diagram of scorpion venom derived peptide FFG-18;
FIG. 3 shows the bactericidal timeliness curve of the scorpion venom derived peptide FFG-18 acting on Escherichia coli;
FIG. 4 shows the result of evaluation of hemolytic toxicity of the scorpion venom derived peptide FFG-18;
FIG. 5 shows the cytotoxicity evaluation results of the scorpion venom derived peptide FFG-18;
FIG. 6 shows a transmission electron microscope image of the effect of the scorpion venom derived peptide FFG-18 on Escherichia coli;
FIG. 7 shows the modulating effect of the scorpion venom derived peptide FFG-18 on cytokines;
FIG. 8 shows the effect of scorpion venom derived peptide FFG-18 on NF- κB.
Detailed Description
In order to make the technical solutions of the present invention more apparent to those skilled in the art, the following examples will be presented. It should be noted that the following embodiments are only preferred embodiments of the present invention, and the scope of the present invention is not limited to the following embodiments, and any modifications, substitutions, and combinations made without departing from the spirit and principles of the present invention are included in the scope of the present invention.
The starting materials, reagents or apparatus used in the following examples are all available from conventional commercial sources or may be obtained by methods known in the art unless otherwise specified.
Synthesis of the polypeptide: the polypeptides used in the experiment are synthesized and purified by the laboratory, the synthesis method is a solid phase synthesis method, and the purity of the purified polypeptides is more than 95% through HPLC detection.
The required experimental materials are:
standard strains E.coli ATCC25922 (Escherichia coli), S.aureus ATCC12600 (Staphylococcus aureus), P.aeromonas ATCC25853 (Pseudomonas aeruginosa) were all purchased from the Guangdong province microorganism strain collection, escherichia coli clinical isolates were obtained from the Guangdong province Margaret national institute of England, and mouse peritoneal mononuclear macrophages (RAW 264.7) were purchased from the China academy of sciences of Shanghai life sciences cell resource center.
The required experimental reagents are:
complete sets of Fmoc protected amino acids, solid phase resin and condensing agent (HBTU, HOBt), purchased from Gill Biochemical Co., ltd. (Shanghai, china); piperidine was purchased from national pharmaceutical group chemical reagent limited (Shanghai, china); ninhydrin, trifluoroacetic acid, 1, 2-ethanedithiol, toluene sulfide, chromatographically pure acetonitrile, chromatographically pure methanol were purchased from aledine biochemical technologies, inc. (Shanghai, china); bacterial culture media (MH medium, LB medium), agar from the seabo biotechnology company (peninsula china); DMEM high sugar medium, 1640 medium, fetal bovine serum, trypsin, purchased from Gibco (usa); CCK-8 kit was purchased from the same-core chemical institute (Japan); ELISA kits (TNF-alpha, IL-1 beta, IL-8) were purchased from Xinbo biotechnology Co., ltd (Shenzhen, china).
Example 1
This example provides a scorpion venom derived peptide (designated FFG-18) having the amino acid sequence: FFGRLFKLAVKIIPSLFK (SEQ ID NO: 1), the scorpion venom derived peptide polypeptide contains 18 amino acid residues, has an average molecular weight of 2124.65g/mol, a hydrophobicity coefficient of 1.03 and a theoretical isoelectric point pH of 11.73.
The scorpion venom derivative peptide can be applied to the preparation of antibacterial products or anti-inflammatory drugs.
Example 2: synthesis method of scorpion venom derived peptide FFG-18
The scorpion venom derived peptide FFG-18 of example 1 was synthesized using standard Fmoc solid phase synthesis, and had the following steps: weighing 0.1mmol of resin, placing in a reactor, adding DCM to swell and activate, 1.5mL/5min, and washing for 2 times; DMF 1.5mL/10min, 2 washes. Adding 25% piperidine/DMF 2mL/5min; adding 25% piperidine/DMF 2mL/25min; DMF 1mL/2min, 6 times. Setting resin: amino acid: TBTU: HOBt: diea=1:3:3:3:6, proper amount of DMF was added as solvent, and the reaction was performed for 3h.2% ninhydrin solution preparation: weighing 0.3g of ninhydrin, and dissolving with 15mL of absolute ethyl alcohol; sucking more than ten resin into a glass tube by a pipette, washing the resin by DMF and MeOH, adding about 200 mu L of ninhydrin solution, heating for 3-5min, observing the color change of the resin, completely reacting the resin without changing the color of the resin to blue, squeezing out the reaction solution, and washing the resin in the reactor by DMF and MeOH for 2 times respectively. Repeating the above steps to connect the next amino acid. The resin was reacted completely with DMF, DCM: meOH (1:1), DCM, 2 washes each and dried; on ice procedure formulation TFA: EDT: tis: h 2 O (v/v, 100:1:2:2); reacting for 3h; squeezing into precipitant petroleum ether: 10mL of methyl tertiary butyl ether (1:1), centrifuged at 5000rpm for 10min, and the supernatant discarded; adding a precipitant petroleum ether: methyl tertiary butyl ether (1:1) 5mL at 8000-9000rpCentrifuging for 10min, and air drying overnight. The next day, the dried polypeptide is placed in a rotary evaporator and a vacuum drying oven in sequence for pumping, refrigerating and preserving.
The purity of the scorpion venom derived peptide FFG-18 is detected by HPLC, and the detection result is shown in figure 1: after purification, the purity of the scorpion venom derived peptide FFG-18 is more than 95%. The molecular weight of the obtained scorpion venom derived peptide FFG-18 was determined by using a MALDI-TOF MS mass spectrometer, and the detection result is shown in FIG. 2.
Example 3: strain culture conditions and minimum inhibitory concentration determination
Pseudomonas aeruginosa, escherichia coli and clinical isolates thereof were cultured under aerobic conditions using LB medium, staphylococcus aureus was cultured under aerobic conditions using MH medium, the growth temperature of the strains was 37℃and the total number of colonies was determined using the turbidimetric method. The test compounds were diluted to the indicated concentration ranges using strain-corresponding media to formulate 96 Kong Yaomin plates. Taking bacterial suspension in logarithmic growth phase, measuring bacterial concentration and diluting to 1X 10 5 CFU/mL, 100. Mu.L of bacterial suspension was added to each well, multiple groups of drug concentrations were set, incubation was continued in the incubator, and the results were read for 17-20 hours, taking the lowest observed drug concentration without signs of bacterial growth as MIC value. Each set of test results was independently repeated 4 to 6 times. The test results are shown in table 1:
TABLE 1 MIC values for different polypeptide Compounds
As can be seen from the results in Table 1, the modified scorpion venom derived peptide FFG-18 has obviously improved antibacterial activity on several test strains compared with the original scorpion venom monomer, and has the best inhibitory effect on the escherichia coli standard strain and clinical isolate, and the MIC reaches 4 mug/mL. The polypeptide consisting of 18 random amino acids has little antibacterial effect on the test strain in the test range, and proves that the scorpion venom derived peptide FFG-18 provided by the invention is reasonably designed and screened, and has the characteristic of high-activity antibacterial effect.
Example 4: determination of bactericidal timeliness relation of scorpion venom derived peptide FFG-18 on escherichia coli
Diluting bacterial suspension in logarithmic growth phase to 1×10 5 CFU/mL, adding scorpion venom derived peptide FFG-18 with a final concentration of 8 mug/mL, respectively treating 0,2,4,6,8 and 10 hours at 25 ℃ under aerobic conditions, respectively diluting bacterial liquid according to the proportion of 1:10,1:100 and 1:1000, respectively taking 20 mu L of diluted liquid, placing the diluted liquid on an agar culture medium plate, culturing for 24 hours at 37 ℃, and recording the colony number of the plate; and 2 mug/mL gentamicin is used as a control drug.
As is apparent from FIG. 3, the inhibition rate of the scorpion venom derived peptide FFG-18 at the minimum inhibitory concentration on Escherichia coli was much faster than that of gentamicin, and after 6 hours of the action, only 1X 10 was observed in the FFG-18 treated group 2 Colonies, while gentamicin treated groups still had 6X 10 4 Is a colony survival. The sterilization mechanism of FFG-18 is different from the traditional aminoglycoside antibiotics, and the FFG-18 can play a role in rapidly inhibiting bacterial proliferation.
Example 5: evaluation of hemolytic toxicity and cytotoxicity of scorpion venom derived peptide FFG-18
Fresh rabbit blood was centrifuged to obtain packed red blood cells (rotation speed: 10000rpm, time: 1 min), resuspended by gentle blowing with equal volume of PBS, centrifuged (rotation speed: 10000rpm, time: 1 min), and repeatedly washed once. The washed red blood cells were resuspended in PBS to a concentration of 5% for further use. FFG-18 is diluted by PBS to obtain 100 mu L of drug solution with the concentration gradient of 2-125 mu g/mL; 1% Triton X-100 positive control wells (representing 100% hemolysis) and solvent control wells (representing 0% hemolysis) were set. After adding 100. Mu.L of 5% rabbit blood suspension per well and incubating at 37℃for 1 hour, centrifugation (rotation speed: 3000rpm, time: 15 min) was performed and 50. Mu.L of the supernatant was taken in a new 96-well plate and absorbance was measured at 570nm with an enzyme-labeled instrument.
RAW 264.7 cells (5X 10) 4 Well) was inoculated in 96-well plates, after overnight incubation, fresh medium containing 2-125. Mu.g/mL FFG-18 was added and allowed to act at 37℃for 24 hours. After that, 10. Mu.L of CCK-8 (Dojindo, japan) was added to each well and incubation was continued for 2 hours at 37 ℃. Absorbance was measured at 450nm using an enzyme-labeled instrument (Infinite M1000 Pro, tecan, switzerland).Cytotoxicity was assessed by comparing the growth inhibition of cells treated with FFG-18. The inhibition rate of cells of the control group which had not been treated with FFG-18 was set to 0%.
As can be seen from FIGS. 4-5, FFG-18 has low hemolytic toxicity and cytotoxicity, and the hemolysis rate at 100. Mu.g/mL concentration is only 28%; IC for RAW 264.7 cells simultaneously 50 = 68.66 μg/mL, well above its inhibitory concentration. It is therefore believed that FFG-18 has a higher safety index over the normal dosage range of use.
Example 6: transmission electron microscope image of scorpion venom derived peptide FFG-18 acting on escherichia coli
The concentration of the bacterial liquid is regulated to be 1 multiplied by 10 6 CFU/mL, treated with 8. Mu.g/mL FFG-18 for 2 hours. The supernatant was discarded by centrifugation (1000 rpm,5 min), washed twice with PBS buffer, and then treated with 2.5% glutaraldehyde overnight. The solution was rinsed with 0.1M phosphate at least three times for 15 minutes each, and fixed with 1% osmium tetroxide for 2-3 hours. Dehydration conditions: 50% ethanol 15-20 min, 70% ethanol 15-20 min, 90% ethanol and 90% acetone 15-20 min (1:1, v/v). Incubation conditions: the mixture of acetone and embedding liquid (2:1, v/v) was incubated at room temperature for 3-4 hours, and the mixture of acetone and embedding liquid (1:2, v/v) was incubated at 37℃for 2-3 hours. Curing conditions: incubation was carried out at 37℃overnight, 45℃for 12 hours, and at 60℃for 24 hours. After curing, the specimens were sectioned, using 3% uranyl acetate and lead citrate as dyes, observed with a transmission electron microscope and photographed to record the results.
From the result of the transmission electron microscope shown in FIG. 6, after the Escherichia coli is acted by the FFG-18 of 8 mug/mL, compared with a blank control group, the cell membrane is damaged, the edge is dissolved, cytoplasm is absent, obvious gaps are formed between the inner edge and the cell membrane, and a clear vacuole structure is formed inside the cell. This result demonstrates that FFG-18 bactericidal effects can lead to target cell disruption and death through a physical cell membrane disruption effect.
Example 7: scorpion venom derived peptide FFG-18 anti-inflammatory effect evaluation experiment
Enzyme-linked immunosorbent assay: RAW 264.7 cells (5X 10) 4 Well) was inoculated in 96-well plates and grown overnight, fresh medium was added at final concentration of 2,8, 32. Mu.g/mL FFG-18, respectively, after 2 hoursLPS was added to each well at a final concentration of 100 ng/mL. At 37 ℃,5% CO 2 After incubation for 6 hours in the environment, cell culture broth was collected and the levels of TNF-. Alpha., IL-1β and IL-8 in the extracellular fluid were determined according to ELISA kit (Neobioscience, china) and 1. Mu.g/mL Melittin was used as a positive control.
Luciferase reporter gene detection: RAW 264.7 cells stably transfected with NF-. Kappa.B were cultured at 5X 10 4 Wells were seeded in 96-well plates and after overnight cell growth, FFG-18 containing 2,8, 32. Mu.g/mL was added, respectively. After 2 hours, LPS or 1X 10 was added at a final concentration of 100ng/mL 7 CFU/mL of heat-inactivated Escherichia coli was added to each well and incubated at 37℃for 6 hours. The supernatant was then discarded, 100. Mu.L of lysate was added to each well, after incubation for 20 minutes in the absence of light, the cell lysate was transferred to a white opaque plate, 100. Mu.L of chemiluminescent substrate was added to each well, and chemiluminescence was detected by a microplate reader (Infinite M1000 Pro, tecan, swiss) and 10. Mu.g/mL PDTC (one of NF-. Kappa.B inhibitors) was used as a positive control.
Lipopolysaccharide (LPS) was used to induce the production of RAW 264.7 pro-inflammatory cytokines to model cellular inflammation. As can be seen from the ELISA results of FIG. 7, when the concentration of FFG-18 was set to 2,8, 32. Mu.g/mL, the FFG-18 at 8. Mu.g/mL inhibited the expression of cytokines TNF-a, IL-1. Beta. And IL-8, and exhibited a more remarkable anti-inflammatory effect.
Considering that nuclear transcription factor κB (NF- κB) plays an important role in the development of inflammation and signal transduction of various downstream inflammatory factors, we tested the inhibition of NF- κB in RAW 264.7 inflammatory model cells by FFG-18. RAW 264.7 cells encoding NF- κB and luciferase reporter stably transformed plasmids were pre-treated with FFG-18, after 2 hours, cells were induced with LPS and heat inactivated E.coli, respectively, for 6 hours, and finally the inhibition of FFG-18 on NF- κB was assessed by comparing luciferase activities of the dosing group and the control group. The results in FIG. 8 show that FFG-18 at 8 μg/mL significantly inhibited NF- κB expression after the steady-state cells induced inflammatory response by LPS or inactivated bacteria.
Example 8
The present example provides a medicament containing the scorpion venom derived peptide FFG-18 in example 1, wherein the medicament is in the form of injection, and comprises the scorpion venom derived peptide FFG-18 with the mass concentration of 10mg/mL and 0.9% sodium chloride solution as a solvent, and the medicament has antibacterial and anti-inflammatory effects.
The embodiments of the present application have been described in detail above with reference to the accompanying drawings, but the present application is not limited to the above embodiments, and various changes can be made within the knowledge of one of ordinary skill in the art without departing from the spirit of the present application. Furthermore, embodiments of the present application and features of the embodiments may be combined with each other without conflict.
SEQUENCE LISTING
<110> Zhuhai City people Hospital
<120> a scorpion venom derivative peptide and application thereof in preparing antibacterial or anti-inflammatory drugs
<130> 1
<160> 1
<170> PatentIn version 3.5
<210> 1
<211> 18
<212> PRT
<213> artificial sequence
<400> 1
Phe Phe Gly Arg Leu Phe Lys Leu Ala Val Lys Ile Ile Pro Ser Leu
1 5 10 15
Phe Lys
Claims (9)
1. A scorpion venom derived peptide, characterized in that the amino acid sequence of the scorpion venom derived peptide is: FFGRLFKLAVKIIPSLFK.
2. The scorpion venom derived peptide of claim 1, wherein the amino acids in the scorpion venom derived peptide are all L-amino acids.
3. An isolated nucleic acid molecule encoding the scorpion venom derived peptide of claim 1 or 2.
4. A recombinant vector comprising the nucleic acid molecule of claim 3.
5. Use of a scorpion venom derived peptide or a pharmaceutically acceptable salt thereof as claimed in claim 1 or 2 in the manufacture of an antibacterial product, wherein the bacterium is escherichia coli, staphylococcus aureus or pseudomonas aeruginosa.
6. The use of claim 5, wherein the pharmaceutically acceptable salt of the scorpion venom derived peptide comprises at least one of acetate and citrate.
7. An antibacterial agent comprising the scorpion venom derived peptide or a pharmaceutically acceptable salt thereof according to claim 1 or 2, and a pharmaceutically acceptable auxiliary material;
the bacteria which can be inhibited by the antibacterial drug are Escherichia coli, staphylococcus aureus or pseudomonas aeruginosa.
8. Use of a scorpion venom derived peptide or a pharmaceutically acceptable salt thereof as claimed in claim 1 or 2 in the manufacture of an anti-inflammatory medicament.
9. An anti-inflammatory agent comprising the scorpion venom derived peptide or a pharmaceutically acceptable salt thereof as claimed in claim 1 or 2, and a pharmaceutically acceptable adjuvant.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111680590.3A CN114349825B (en) | 2021-12-30 | 2021-12-30 | Scorpion venom derivative peptide and application thereof in preparation of antibacterial or anti-inflammatory drugs |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111680590.3A CN114349825B (en) | 2021-12-30 | 2021-12-30 | Scorpion venom derivative peptide and application thereof in preparation of antibacterial or anti-inflammatory drugs |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114349825A CN114349825A (en) | 2022-04-15 |
| CN114349825B true CN114349825B (en) | 2023-06-23 |
Family
ID=81105324
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111680590.3A Active CN114349825B (en) | 2021-12-30 | 2021-12-30 | Scorpion venom derivative peptide and application thereof in preparation of antibacterial or anti-inflammatory drugs |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114349825B (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101063103A (en) * | 2007-04-26 | 2007-10-31 | 武汉大学 | Hainan sporic scorpion antibiotic and preparation method and application |
| CN101063102A (en) * | 2007-04-26 | 2007-10-31 | 武汉大学 | East-Asia scorpion antibiotic peptide gene and preparation method and application |
| CN103865935A (en) * | 2014-03-31 | 2014-06-18 | 中国地质大学（武汉） | Opistophthalmus glabrifrons antibacterial peptide gene, antibacterial peptide Os_116, homologous structure antibacterial peptide of antibacterial peptide Os_116, preparation method and application |
| WO2015007216A1 (en) * | 2013-07-17 | 2015-01-22 | 武汉摩尔生物科技有限公司 | Polypeptide, dna molecule encoding the polypeptide, vector, preparation method and use |
| CN111423501A (en) * | 2020-03-30 | 2020-07-17 | 东北农业大学 | Antibacterial peptide derived from scorpion venom as well as preparation method and application thereof |
-
2021
- 2021-12-30 CN CN202111680590.3A patent/CN114349825B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101063103A (en) * | 2007-04-26 | 2007-10-31 | 武汉大学 | Hainan sporic scorpion antibiotic and preparation method and application |
| CN101063102A (en) * | 2007-04-26 | 2007-10-31 | 武汉大学 | East-Asia scorpion antibiotic peptide gene and preparation method and application |
| WO2015007216A1 (en) * | 2013-07-17 | 2015-01-22 | 武汉摩尔生物科技有限公司 | Polypeptide, dna molecule encoding the polypeptide, vector, preparation method and use |
| CN103865935A (en) * | 2014-03-31 | 2014-06-18 | 中国地质大学（武汉） | Opistophthalmus glabrifrons antibacterial peptide gene, antibacterial peptide Os_116, homologous structure antibacterial peptide of antibacterial peptide Os_116, preparation method and application |
| CN111423501A (en) * | 2020-03-30 | 2020-07-17 | 东北农业大学 | Antibacterial peptide derived from scorpion venom as well as preparation method and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| 刘霞 ; 任可可 ; 白占涛 ; .陕北产东亚钳蝎毒液和体液的体外抑菌活性.延安大学学报(自然科学版).2016,(02),第55-58页. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114349825A (en) | 2022-04-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110903347A (en) | Antibacterial peptide L7 and application thereof | |
| CN113307855A (en) | Scylla paramamosain antibacterial polypeptide Spampcin56-86And uses thereof | |
| CN111363010B (en) | Symmetrical short-sequence antibacterial peptide analogue and application thereof | |
| CN114853865B (en) | Modified antibacterial peptide dsNCM1 and application thereof | |
| CN112625107A (en) | Modified antibacterial peptide C-CM8 of tortoise green antibacterial peptide, and preparation method and application thereof | |
| CN110498848B (en) | Melittin variant and application thereof | |
| CN106749559B (en) | Antibacterial peptide based on cell-penetrating peptide Tat (49-57) and synthesis method thereof | |
| CN112625108A (en) | Modified antibacterial peptide C-CM5 of tortoise green antibacterial peptide, and preparation method and application thereof | |
| US9090655B2 (en) | Low hemolytic antimicrobial peptide, pharmaceutical composition and use thereof | |
| CN111253474B (en) | Antibacterial peptide RG-27 and application thereof | |
| JP2013521330A (en) | Peptide compounds useful as antibacterial agents | |
| CN114349825B (en) | Scorpion venom derivative peptide and application thereof in preparation of antibacterial or anti-inflammatory drugs | |
| CN110054664B (en) | Side chain fatty acid modified antibacterial peptide analogue containing D-type amino acid and synthesis and application thereof | |
| CN114891068B (en) | Antibacterial peptide GK18 and application thereof | |
| CN115043925B (en) | Modified antibacterial peptide oNCM and application thereof | |
| CN115785220B (en) | Tryptophan-enriched antibacterial peptide with high protease stability and preparation method and application thereof | |
| CN113999297B (en) | Antibacterial peptide hrNCM and preparation method and application thereof | |
| Nan et al. | Antimicrobial and anti-inflammatory activities of a Leu/Lys-rich antimicrobial peptide with Phe-peptoid residues | |
| CN110551191B (en) | Melittin with low hemolytic activity and application thereof | |
| CN109602894B (en) | Application of cecropin derived peptide | |
| US10100086B2 (en) | Peptide and uses thereof | |
| CN107827988B (en) | Chimeric transmembrane antibacterial peptide T11N2 and application thereof | |
| CN118005739B (en) | Polypeptide APH229 of drug-resistant strain, preparation method and application thereof | |
| CN111533788B (en) | Cell-penetrating antibacterial peptide targeting streptococcus agalactiae and preparation method and application thereof | |
| CN115925802B (en) | Antibacterial peptide with enhanced antibacterial activity and serum stability and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |