CN1597698A - Cation antibacterial peptide - Google Patents
Cation antibacterial peptide Download PDFInfo
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- CN1597698A CN1597698A CN 200410020077 CN200410020077A CN1597698A CN 1597698 A CN1597698 A CN 1597698A CN 200410020077 CN200410020077 CN 200410020077 CN 200410020077 A CN200410020077 A CN 200410020077A CN 1597698 A CN1597698 A CN 1597698A
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- xaa
- antibacterial peptide
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- ile
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Abstract
An anionic antibacterial peptide used as the medicine for treating the diseases caused by gram-positive and gram-negative bacteria and fungus is composed of an alkaline amino acid cluster consisting of 2-4 continuous alkaline amino acids and a neutral amino acid fragment consisting of 8-9 neutral amino acids. It has high antibacterial activity and low hemolytic activity.
Description
[technical field]
The present invention relates to antibacterial peptide, the specifically segment and the analogue polypeptide thereof of mellitin (melittin), and cationic antibacterial peptide is as the application of antibiotic.
[background technology]
Countless life has been saved in the use of antibiotic, and people's predicted life was increased more than 10 years.But along with being extensive use of of antibiotic, bacterium has also produced resistance to it gradually, forces people constantly to seek new antibiotic.Though the antibiotic of present clinical use (comprising inactive) has 150 kinds more than approximately, but they mostly belong to the 6 big classes of finalizing the design before more than 30 year, newly Yan Zhi antibiotic is generally all within this 6 big class scope, with existing antibiotic cross resistance is often arranged, the new antibiotic of seeking the overriding resistance bacterium becomes more and more difficult.Drug-resistance of bacteria has become very serious now, and some superbacterias comprise that to all antibiotic of present use being called as antibiotic last-ditch vancomycin has resistance (A.H.Wong, R.P.Wenzel, M.B.Edmond, Am.J.Infect.Control, 2000,28,277-281).Because it is of common occurrence in recent years in the case at fatal position that the infection that drug-resistant bacteria causes causes the people, some controlled originally transmissible diseases are staged a comeback again in recent years.Therefore seek novel type, that bacterium is difficult for producing chemical sproof antibiotic to it is extremely urgent.
There are many antibacterial peptides in various biologies (comprising plant, the animal and human's class) body, be used for tackling bacterium, virus and fungi etc., this is the most ancient but still effectively so far broad-spectrum antibiotics of a class, illustrate that bacterium is difficult for it is produced resistance (M.Zasloff, Nature, 2002,415,389-395).This class polypeptide also claims cationic antibacterial peptide because of all containing more basic aminoacids.Natural cationic antibacterial peptide is of a great variety, found above 500 kinds; Structure is different, size from 14, five amino acid is to more or less a hundred amino acid, has the line style peptide also to have to contain two sulphur strong or do not contain the strong cyclic peptide of two sulphur.Because unique antibiotic mechanism, bacterium are difficult for it is produced resistance.Therefore this class polypeptide and analogue thereof have caused people's great attention as the antibiotic of new generation of overriding resistance bacterium.Though from various biologies, found to surpass 500 kinds of antibacterial peptides, directly utilized natural antibacterial peptide seldom as the trial of antibiotic.Major cause has: some natural antibacterial peptide anti-microbial activity is lower; The long thereby synthetic cost height of some natural antibacterial peptide peptide chains; Also have some natural antibacterial peptides except that its anti-microbial activity, also have very strong side effects such as haemolysis.Therefore people often attempt the analogue of synthesis of natural antibacterial peptide, its objective is: improve anti-microbial activity, reduce the haemolysis side effect, reduce the total number of atnino acid of peptide.
In numerous natural antibacterial peptides, the mellitin (melittin) that is present in the meltittin venom gets more and more people's extensive concerning.The polypeptide that mellitin is made up of 26 amino acid, its aminoacid sequence is:
Gly-Ile-Gly-Ala-Val-Leu-Lys-Val-Leu-The-The-Gly-Leu-
Pro-Ala-Leu-Ile-Ser-Trp-Ile-Lys-Lys-Arg-Arg-Gln-GIn-NH
2
Mellitin all has very strong restraining effect to Gram-positive and negative bacterium, also has antimycotic, antitumor, radioprotective, the arthritic effect of resisting rheumatoid disease.But mellitin also has very strong haemolysis side effect simultaneously, and this makes mellitin be subjected to great obstruction as the application of antibiotic.
[summary of the invention]
One object of the present invention is just at the shortcoming of above-mentioned mellitin as antibiotic, and pass through chemical method, by synthetic mellitin segment, the pulsating replacement of mellitin, the test of putting upside down the order of amino-acid residue in the peptide chain, synthesized one group and had higher anti-microbial activity, low hemolytic activity and the mellitin analogue long, i.e. cationic antibacterial peptide than short chain.Another object of the present invention is that this cationic antibacterial peptide is applied to prepare in the medicine for the treatment of gram-positive microorganism, Gram-negative bacteria and fungi infestation.
The invention provides one group of cationic antibacterial peptide, it is characterized in that each antibacterial peptide is made up of two parts: the basic aminoacids that a part is formed for 2-4 successive basic aminoacids bunch, another part is 8 or 9 neutral amino acids segments that neutral amino acids is formed, basic aminoacids bunch both can be positioned at the C end, also can be positioned at the N end.
The present invention discovers, the segment (Mel (13-24)) of the segment (Mel (12-23)) of the segment of the 12-24 residue of mellitin (Mel (12-24)), 12-23 residue, the segment (Mel (12-22)) of 12-22 residue, 13-24 residue, the segment (Mel (13-23)) of 13-23 residue and the segment (Mel (13-22)) of 13-22 residue, by the pulsating C end of these mellitins of chemical method synthetic both can be free carboxyl group, can be amide group also, be more prone to amide group.Its structure sees Table 1.Compare with mellitin, some slightly descends the anti-microbial activity of these synthetic peptides, and some change little, and other anti-microbial activity even bigger than going back of mellitin; But their haemolysis side effect reduces significantly; Their chain length reduces to 10-13 amino-acid residue by 26 amino-acid residues.The reducing of chain length reduces its synthetic cost widely.
The pulsating aminoacid sequence of table 1 mellitin
| Code name | Aminoacid sequence |
| Mel(12-24) Mel(12-23) Mel(12-22) Mel(13-24) Mel(13-23) Mel(13-22) | ?Gly-Leu-Pro-Ala-Leu-Ile-Ser-Trp-Ile-Lys-Lys-Arg-Arg-NH 2?Gly-Leu-Pro-Ala-Leu-Ile-Ser-Trp-Ile-Lys-Lys-Arg-NH 2?Gly-Leu-Pro-Ala-Leu-Ile-Ser-Trp-Ile-Lys-Lys-NH 2?Leu-Pro-Ala-Leu-Ile-Ser-Trp-Ile-Lys-Lys-Arg-Arg-NH 2?Leu-Pro-Ala-Leu-Ile-Ser-Trp-Ile-Lys-Lys-Arg-NH 2?Leu-Pro-Ala-Leu-Ile-Ser-Trp-Ile-Lys-Lys-NH 2 |
Another discovery of the present invention is, when with the single replacement of other amino acid or simultaneously in a plurality of replacement peptide chains when 8 (for Mel (13-24), Mel (13-23), the Mel (13-22)) in 2 Lys left sides or 9 (for Mel (12-24), Mel (12-23), Mel (12-22)) amino acid, the specificity of the kind of the amino-acid residue on each is not strong.On the whole, these 8 or 9 amino acid whose hydrophobicitys are bigger, and then its anti-microbial activity is bigger.Keep higher anti-microbial activity, 4 hydrophobic amino acids will be arranged at least until all being hydrophobic amino acid in these 8 or 9 amino-acid residues, hydrophobic amino acid comprises Leu, Ile, Trp, Phe, Tyr or Val here.Replace one or more anti-microbial activities that all can reduce peptide significantly in these 8 or 9 amino acid with acidic amino acid (as Glu or Asp).As mentioned above, the quantity that increases the hydrophobic amino acid in the neutral amino acids segment can increase its anti-microbial activity, but the solid phase synthesis yield of polypeptide can reduce, and the gained peptide is water-soluble also lower.
Another discovery of the present invention is that when the Lys of the C of these peptides end replaced with Lys with Arg replacement or Arg, the anti-microbial activity of its peptide changed little.
Compare with mellitin, the hemolytic activity of the analogue of the pulsating replacement of above-mentioned these mellitins all significantly reduces.
Another discovery of the present invention is that (the original C end was held as N, and original N end is held as C when the sequence of the synthetic peptide of above-mentioned these was put upside down, synthetic peptide like this is the inverted sequence peptide, the C end of inverted sequence peptide still is an amide group), its anti-microbial activity changes little, and its hemolytic activity is lower.
According to the aminoacid sequence of these synthetic peptides, the constitutional features that sums up them is: these synthetic peptides are made up of two parts, and a part is the basic aminoacids formed of 2-4 successive basic aminoacids bunch, and it can be made up of any basic aminoacids; Another part is 8 or 9 neutral amino acids segments that neutral amino acids is formed.Replace the pulsating test of neutral amino acids and show, contain 4 hydrophobic amino acids in the neutral amino acids segment at least, comprise Leu, Ile, Trp, Phe, Tyr or Val; Other is any neutral amino acids, as Gly, Ala, Ser, Thr, Pro, Asn or Gln.Basic aminoacids bunch both can be positioned at the C end, also can be positioned at the N end, and the C end of peptide is amide group.The aminoacid sequence of some (but being not limited to these) inverted sequence peptides sees Table 2.
The aminoacid sequence of some inverted sequence peptides of table 2
| Code name | Aminoacid sequence |
| RetroMel(12-24) RetroMel(12-23) RetroMel(12-22) RetroMel(13-24) RetroMel(13-23) RetroMel(13-22) | ?Arg-Arg-Lys-Lys-Ile-Trp-Ser-Ile-Leu-Ala-Pro-Leu-Gly-NH 2?Arg-Lys-Lys-Ile-Trp-Ser-Ile-Leu-Ala-Pro-Leu-Gly-NH 2?Lys-Lys-Ile-Trp-Ser-Ile-Leu-Ala-Pro-Leu-Gly-NH 2?Arg-Arg-Lys-Lys-Ile-Trp-Ser-Ile-Leu-Ala-Pro-Leu-NH 2?Arg-Lys-Lys-Ile-Trp-Ser-Ile-Leu-Ala-Pro-Leu-NH 2?Lys-Lys-Ile-Trp-Ser-Ile-Leu-Ala-Pro-Leu-NH 2 |
The present invention is in its preferred version, when the arbitrary amino-acid residue in the above-mentioned antibacterial peptide (comprising positive sequence and inverted sequence) (replaces one or more residues during by corresponding D type aminoacid replacement in the polypeptide, until whole residues by corresponding D type aminoacid replacement), its anti-microbial activity is constant substantially, and hemolytic activity can further reduce.Another advantage with D type aminoacid replacement is that the speed that its polypeptide is degraded in human body can reduce greatly.
Antimicrobial polypeptide of the present invention can adopt solid-phase polypeptide synthesis method or liquid phase polypeptide synthesis to synthesize.
Cationic antibacterial peptide of the present invention is used for being applied to prepare in the medicine for the treatment of gram-positive microorganism, Gram-negative bacteria and fungi infestation.
[embodiment]
EXAMPLE l
With Mel (12-24) is example, and the method that adopts the synthetic polypeptide of solid phase method is described:
Add 1g Rinkamide mbha resin (0.74mmol N/g) in 50ml side band arm, arm have the round-bottomed flask of sand plate filter core, add 15ml DMF swelling 10 minutes, suction filtration removes solvent.Add 15ml 20% piperidines/DMF solution then, stirred suction filtration 30 minutes.With DMF washing resin 6 times, suction filtration removes solvent.(Pbf is 2 to add 1.44g (2.22mmol) Fmoc-Arg (Pbf)-OH in reaction flask; 2; 4; 6; 7-pentamethyl-Dihydrobenzofuranes-5-alkylsulfonyl), 0.46g (2.22mmol) DCC (dicyclohexylcarbodiimide) and 0.30g (2.22mmol) HOBt (1-hydroxy benzo triazole) and 15ml DMF, stir under the room temperature and carry out condensation reaction.React the resin that takes a morsel after 2 hours and carry out the triketohydrindene hydrate coloring test, the result shows that condensation reaction is complete.Suction filtration is used DMF washing resin 6 times, and suction filtration removes solvent.First (protection) amino acid of C end is connected on the resin.The back then connects hold by C second to last (protection) amino acid, and the operation above reactions steps repeats promptly from 20% piperidines/DMF solution deprotection, is only washed with DMF after arriving the triketohydrindene hydrate coloring test.If it is incomplete that the triketohydrindene hydrate coloring test shows condensation, then can prolong the condensation time until fully.All amino acid whose alpha-amino groups all with the Fmoc protection, have the amino acid of Side chain protective group to be: Fmoc-Lys (Boc)-OH, Fmoc-Ser (tBu)-OH (tBu is the tertiary butyl), Fmoc-Trp (Boc)-OH.Last resin methanol wash three times, vacuum-drying.
The above-mentioned resin that has connect peptide is joined in the 50ml round-bottomed flask, add 15ml reagent K (trifluoroacetic acid/water/phenol/thioanisole/1,2-dimercapto ethylene glycol=82.5/5/5/5/2.5, volume ratio), stirred 4 hours under the room temperature.Filter, collect filtrate.With give a baby a bath on the third day after its birth time resin of trifluoroacetic acid.Merging filtrate revolves steaming with filtrate and removes most trifluoroacetic acid, and adding volume is 8-10 ether sedimentation polypeptide doubly, puts into refrigerator overnight.Centrifugal, remove ether, vacuum-drying obtains the thick peptide of Mel (12-24).
The thick peptide of the Mel that 200mg is above-mentioned (12-24) is dissolved in the 2ml pure water, with preparation type reversed-phase HPLC purifying, chromatographic column is μ Bondapak C18 (19mm * 300mm), moving phase is: trifluoroacetic acid/acetonitrile/water (0.1/32/68, volume ratio), flow velocity is 12ml/ minute, and the collection retention volume is 16 minutes a main peak.Collect liquid and concentrate through rotary evaporation, lyophilize gets the pure peptide of Mel (12-24) then.The mass spectrum of pure peptide shows that its molecular weight is 1537, conforms to calculated value.
Embodiment 2
The mensuration of the anti-microbial activity of polypeptide: be～lmg/ml polypeptide sample solution with the sterile saline compound concentration.Test is inoculated in nutrient broth respectively with bacterial classification, cultivates 18-24 hour in 370C, faces with preceding and does dilution in 1: 105 with physiological saline, gets 12 microbial culture pipes and numbering, adds nutrient broth 1.8ml in first pipe, all adds 1.0ml meat soup in all the other 11 pipes.Add the 0.2ml polypeptide solution in the 1st pipe, take out 1.0ml behind the mixing and join in second pipe, so repeatedly, be diluted to the 12nd pipe successively, discard the remaining lml of lml, every pipe adds is ready for bacterium liquid 0.2ml, and jolting is even gently, puts 370C and cultivates 18-24 hour, observes.Cultivate the back culture tube and present clarification, this pipe asepsis growth is thought in still clarification after the jolting; Culture tube presents muddy state and shows that bacteria growing is arranged.Find out the culture tube of minimum peptide concentration from each pipe of asepsis growth, the peptide concentration of this pipe is the minimum sample concentration (MIC) of bacteria growing inhibiting.Table 3 is the minimum inhibitory concentration of part antibacterial peptide to several bacteriums.
Table 3 antibacterial peptide minimum inhibitory concentration MIC (μ g/ml)
| Polypeptide * | Gold Portugal bacterium eel grass bacillus intestinal bacteria Candida albicans |
| melittin Mel(12-24) Mel(12-23) Mel(12-22) Mel(13-24) Mel(13-22) Mel(12-24，L 1) Mel(12-24，S 6) Mel(13-22，L 3) Mel(12-24，F 8) Mel(12-23，dL 4dI 9) RetroMel(12-24) RetroMel(12-23) RetroMel(12-22) RetroMel(13-24) RetroMel(12-24，L 11) RetroMel(12-24，L 11L 13) RetroMel(13-23，L 6) | ?0.72?????1.4???????12????????24 ?2.6??????1.3???????21????????42 ?5.2??????5.2???????42????????84 ?10???????5.2???????42????????84 ?2.6??????2.6???????21????????84 ?10???????10????????21????????42 ?0.65?????1.3???????5.2???????10 ?7.3??????29????????117???????230 ?5.2??????2.6???????10????????21 ?3.0??????6.0???????24????????48 ?5.2??????5.2???????21????????42 ?2.9??????1.5???????5.4???????10 ?6.6??????6.6???????6.6???????13 ?25???????13????????52????????100 ?2.9??????1.5???????2.9???????6.0 ?1.2??????2.4???????9.6???????20 ?0.32?????0.32??????1.3???????5.2 ?0.6??????1.2???????4.8???????10 |
* the aminoacid replacement that this polypeptide of numeral is represented by this capitalization in the position shown in the numeral is marked in the capital letter master tape upper right corner in the bracket, and d represents D type amino acid.As Mel (12-24, L
1) replaced by Leu (single-letter is abbreviated as L) for the amino acid on the 1st of Mel (12-24); RetroMel (12-24, L for another example
1L
3) replaced by Leu respectively for the 11st and the 13rd of RetroMel (12-24).Other single-letter is abbreviated as in the table: S-Ser; F-Phe.Mlc is littler in the table, and then anti-microbial activity is higher.
Embodiment 3
The mensuration of the hemolytic activity of polypeptide: human red cell is suspended in the phosphate buffered saline buffer (pH=7.4), obtains erythrocyte suspension (5%v/v).Polypeptide is dissolved in the phosphate buffered saline buffer, be made into about 5mg/ml storing solution, get 14 1.5ml centrifuge tubes, in first centrifuge tube, add 1ml polypeptide storing solution, add the 0.5ml phosphate buffered saline buffer in all the other each pipes, taking out 0.5ml polypeptide storing solution from first pipe adds in second pipe, mix with micro-mixed instrument, taking out 0.5ml solution again from second pipe adds in the 3rd pipe and mixes, by that analogy, promptly be diluted to the 14th pipe successively, discard 0.5ml with the sesquialter dilution method, each pipe is added 5% erythrocyte suspension that 0.5ml prepares to final volume 1.0ml, shake up gently, in 37 ℃ of thermostat containers the insulation 60 minutes after in 4000 rev/mins centrifugal 10 minutes, get supernatant liquor colorimetric under 414nm, being suspended in the phosphate buffered saline buffer with erythrocyte is blank, and being suspended among the 1%TritonX-100 with erythrocyte is 100% haemolysis.Percentage of hemolysis is calculated with following formula:
The definition percentage of hemolysis is that 50% o'clock peptide concentration is homolytic dose (HC
50).Table 4 is the hemolytic activity of part of polypeptide.
The hemolytic activity HC of table 4 antibacterial peptide
50
| Polypeptide | ????HC 50(μg/ml) * |
| melittin Mel(12-24) Mel(12-23) Mel(12-22) Mel(13-24) Mel(13-22) Mel(12-24，L 1) Mel(12-24，S 6) Mel(13-22，L 3) Mel(12-24，F 8) Mel(12-23，dL 4dI 9) RetroMel(12-24) RetroMel(12-23) RetroMel(12-22) RetroMel(13-24) RetroMel(12-24，L 11) RetroMel(12-24，L 11L 13) RetroMel(13-23，L 6) | ????5.0 ????2200 ????＞2500 ????＞2500 ????＞2500 ????＞2500 ????1800 ????＞2500 ????140 ????1900 ????＞2500 ????＞2500 ????＞2500 ????＞2500 ????＞2500 ????＞2500 ????＞2500 ????＞2500 |
*HC
50Littler, then hemolytic activity is higher.
Sequence table
＜110〉Nankai University
＜120〉cationic antibacterial peptide
<160>24
<210>1
<211>10
<212>PRT
＜213〉apis mellifera (Apis mellifera)
<220>
<221>CHAIN
＜223〉the 13-22 residue segment of mellitin
<400>1
Leu?Pro?Ala?Leu?Ile?Ser?Trp?Ile?Lys?Lys
1???????????5??????????????10
<210>2
<211>11
<212>PRT
＜213〉apis mellifera (Apis mellifera)
<220>
<221>CHAIN
＜223〉the 13-23 residue segment of mellitin
<400>2
Leu?Pro?Ala?Leu?Ile?Ser?Trp?Ile?Lys?Lys?Arg
1???????????5??????????????10
<210>3
<211>12
<212>PRT
＜213〉apis mellifera (Apis mellifera)
<220>
<221>CHAIN
＜223〉the 13-24 residue segment of mellitin
<400>3
Leu?Pro?Ala?Leu?Ile?Ser?Trp?Ile?Lys?Lys?Arg?Arg
1???????????5??????????????10
<210>4
Sequence table
<211>11
<212>PRT
＜213〉apis mellifera (Apis mellifera)
<220>
<221>CHAIN
＜223〉the 12-22 residue segment of mellitin
<400>4
Gly?Leu?Pro?Ala?Leu?Ile?Ser?Trp?Ile?Lys?Lys
1???????????5??????????????10
<210>5
<211>12
<212>PRT
＜213〉apis mellifera (Apis mellifera)
<220>
<221>CHAIN
＜223〉the 12-23 residue segment of mellitin
<400>5
Gly?Leu?Pro?Ala?Leu?Ile?Ser?Trp?Ile?Lys?Lys?Arg
1???????????5??????????????10
<210>6
<211>13
<212>PRT
＜213〉apis mellifera (Apis mellifera)
<220>
<221>CHAIN
＜223〉the 12-24 residue segment of mellitin
<400>6
Gly?Leu?Pro?Ala?Leu?Ile?Ser?Trp?Ile?Lys?Lys?Arg?Arg
1???????????5??????????????10
<210>7
<211>10
<212>PRT
＜213〉artificial sequence
<220>
<221>CHAIN
＜223〉the pulsating inverted sequence peptide of the 13-22 residue of mellitin
<400>7
Sequence table
Lys?Lys?Ile?Trp?Ser?Ile?Leu?Ala?Pro?Leu
1???????????5??????????????10
<210>8
<211>11
<212>PRT
＜213〉artificial sequence
<220>
<221>CHAIN
＜223〉the pulsating inverted sequence peptide of the 13-23 residue of mellitin
<400>8
Arg?Lys?Lys?Ile?Trp?Ser?Ile?Leu?Ala?Pro?Leu
1???????????????5??????????????10
<210>9
<211>12
<212>PRT
＜213〉artificial sequence
<220>
<221>CHAIN
＜223〉the pulsating inverted sequence peptide of the 13-24 residue of mellitin
<400>9
Arg?Arg?Lys?Lys?Ile?Trp?Ser?Ile?Leu?Ala?Pro?Leu
1????????????5??????????????10
<210>10
<211>11
<212>PRT
＜213〉artificial sequence
<220>
<221>CHAIN
＜223〉the pulsating inverted sequence peptide of the 12-22 residue of mellitin
<400>10
Lys?Lys?Ile?Trp?Ser?Ile?Leu?Ala?Pro?Leu?Gly
1???????????????5??????????????10
<210>11
<211>12
<212>PRT
＜213〉artificial sequence
<220>
Sequence table
<221>CHAIN
＜223〉the pulsating inverted sequence peptide of the 12-23 residue of mellitin
<400>11
Arg?Lys?Lys?Ile?Trp?Ser?Ile?Leu?Ala?Pro?Leu?Gly
1???????????????5??????????????10
<210>12
<211>13
<212>PRT
＜213〉artificial sequence
<220>
<221>CHAIN
＜223〉the pulsating inverted sequence peptide of the 12-24 residue of mellitin
<400>12
Arg?Arg?Lys?Lys?Ile?Trp?Ser?Ile?Leu?Ala?Pro?Leu?Gly
1???????????????5??????????????????10
<210>13
<211>10
<212>PRT
＜213〉artificial sequence
<220>
<221>VARIANT
<222>(1，2，3，4，5，6，7，8)
＜223〉Xaa=Leu or Ile or Trp or Phe or Tyr or Val or Gly or Ala or Ser or Thr or Pro or Asn or Gln, wherein at least 4 Xaa are Leu or Ile or Trp or Phe or Tyr or Val
<222>(9，10)
＜223〉Xaa=Arg or Lys
<400>13
Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa
1???????????????5??????????????????10
<210>14
<211>11
<212>PRT
＜213〉artificial sequence
<220>
<221>VARIANT
<222>(1，2，3，4，5，6，7，8)
＜223〉Xaa=Leu or Ile or Trp or Phe or Tyr or Val or Gly or Ala or Ser or Thr or Pro or Asn or Gln, wherein at least 4 Xaa are Leu or Ile or Trp or Phe or Tyr or Val
<222>(9，10，11)
Sequence table
＜223〉Xaa=Arg or Lys
<400>14
Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa
1???????????????5??????????????????10
<210>15
<211>12
<212>PRT
＜213〉artificial sequence
<220>
<221>VARIANT
<222>(1，2，3，4，5，6，7，8)
＜223〉Xaa=Leu or Ile or Trp or Phe or Tyr or Val or Gly or Ala or Ser or Thr or Pro or Ash or Gln, wherein at least 4 Xaa are Leu or Ile or Trp or Phe or Tyr or Val
<222>(9，10，11，12)
＜223〉Xaa=Arg or Lys
<400>15
Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa
1???????????????5??????????????????10
<210>16
<211>11
<212>PRT
＜213〉artificial sequence
<220>
<221>VARIANT
<222>(1，2，3，4，5，6，7，8，9)
＜223〉Xaa=Leu or Ile or Trp or Phe or Tyr or Val or Gly or Ala or Ser or Thr or Pro or Asn or Gln, wherein at least 4 Xaa are Leu or Ile or Trp or Phe or Tyr or Val
<222>(10，11)
＜223〉Xaa=Arg or Lys
<400>16
Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa
1???????????????5??????????????????10
<210>17
<211>12
<212>PRT
＜213〉artificial sequence
<220>
<221>VARIANT
<222>(1，2，3，4，5，6，7，8，9)
Sequence table
＜223〉Xaa=Leu or Ile or Trp or Phe or Tyr or Val or Gly or Ala or Ser or Thr or Pro or Asn or Gln, wherein at least 4 Xaa are Leu or Ile or Trp or Phe or Tyr or Val
<222>(10，11，12)
＜223〉Xaa=Arg or Lys
<400>17
Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa
1???????????????5??????????????????10
<210>18
<211>13
<212>PRT
＜213〉artificial sequence
<220>
<221>VARIANT
<222>(1，2，3，4，5，6，7，8，9)
＜223〉Xaa=Leu or Ile or Trp or Phe or Tyr or Val or Gly or Ala or Ser or Thr or Pro or Asn or Gln, wherein at least 4 Xaa are Leu or Ile or Trp or Phe or Tyr or Val
<222>(10，11，12，13)
＜223〉Xaa=Arg or Lys
<400>18
Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa
1???????????????5??????????????????10
<210>19
<211>10
<212>PRT
＜213〉artificial sequence
<220>
<221>VARIANT
<222>(1，2)
＜223〉Xaa=Arg or Lys
<222>(3，4，5，6，7，8，9，10)
＜223〉Xaa=Leu or Ile or Trp or Phe or Tyr or Val or Gly or Ala or Ser or Thr or Pro or Asn or Gln, wherein at least 4 Xaa are Leu or Ile or Trp or Phe or Tyr or Val
<400>19
Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa
1???????????????5??????????????????10
<210>20
<211>11
<212>PRT
＜213〉artificial sequence
Sequence table
<220>
<221>VARIANT
<222>(1，2，3)
＜223〉Xaa=Arg or Lys
<222>(4，5，6，7，8，9，10，11)
＜223〉Xaa=Leu or Ile or Trp or Phe or Tyr or Val or Gly or Ala or Ser or Thr or Pro or Asn or Gln, wherein at least 4 Xaa are Leu or Ile or Trp or Phe or Tyr or Val
<400>20
Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa
1???????????????5??????????????????10
<210>21
<211>12
<212>PRT
＜213〉artificial sequence
<220>
<221>VARIANT
<222>(1，2，3，4)
＜223〉Xaa=Arg or Lys
<222>(5，6，7，8，9，10，11，12)
＜223〉Xaa=Leu or Ile or Trp or Phe or Tyr or Val or Gly or Ala or Ser or Thr or Pro or Asn or Gln, wherein at least 4 Xaa are Leu or Ile or Trp or Phe or Tyr or Val
<400>21
Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa
1???????????????5??????????????????10
<210>22
<211>11
<212>PRT
＜213〉artificial sequence
<220>
<221>VARIANT
<222>(1，2)
＜223〉Xaa=Arg or Lys
<222>(3，4，5，6，7，8，9，10，11)
＜223〉Xaa=Leu or Ile or Trp or Phe or Tyr or Val or Gly or Ala or Ser or Thr or Pro or Asn or Gln, wherein at least 4 Xaa are Leu or Ile or Trp or Phe or Tyr or Val
<400>22
Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa
1???????????????5??????????????????10
<210>23
<211>12
Sequence table
<212>PRT
＜213〉artificial sequence
<220>
<221>VARIANT
<222>(1，2，3)
＜223〉Xaa=Arg or Lys
<222>(4，5，6，7，8，9，11，12)
＜223〉Xaa=Leu or Ile or Trp or Phe or Tyr or Val or Gly or Ala or Ser or Thr or Pro or Asn or Gln, wherein at least 4 Xaa are Leu or Ile or Trp or Phe or Tyr or Val
<400>23
Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa
1???????????????5??????????????????10
<210>24
<211>13
<212>PRT
＜213〉artificial sequence
<220>
<221>VARIANT
<222>(1，2，3，4)
＜223〉Xaa=Arg or Lys
<222>(5，6，7，8，9，10，11，12，13)
＜223〉Xaa=Leu or Ile or Trp or Phe or Tyr or Val or Gly or Ala or Ser or Thr or Pro or Asn or Gln, wherein at least 4 Xaa are Leu or Ile or Trp or Phe or Tyr or Val
<400>24
Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa?Xaa
1???????????????5??????????????????10
Claims (9)
1. one group of cationic antibacterial peptide, it is characterized in that each antibacterial peptide is made up of two parts: the basic aminoacids that a part is formed for 2-4 successive basic aminoacids bunch, another part is 8 or 9 neutral amino acids segments that neutral amino acids is formed, basic aminoacids bunch both can be positioned at the C end, also can be positioned at the N end.
2. according to the described antibacterial peptide of claim 1, it is characterized in that containing in the described neutral amino acids segment at least 4 hydrophobic amino acids until being hydrophobic amino acid all, other residue except that hydrophobic amino acid is any other neutral amino acids.
3. according to the described antibacterial peptide of claim 1, it is characterized in that described basic aminoacids bunch can be made up of any basic aminoacids.
4. according to claim 1 or 2,3 described antibacterial peptides, it is characterized in that in the antibacterial peptide any one or a plurality of can be until whole amino-acid residues by its enantiomorph D type aminoacid replacement.
5. according to the described antibacterial peptide of claim 2, it is characterized in that the hydrophobic amino acid that comprises in the described neutral amino acids segment comprises Leu, Ile, Trp, Phe, Tyr or Val.
6. according to the described antibacterial peptide of claim 2, it is characterized in that other neutral amino acids that comprises in the described neutral amino acids segment comprises Gly, Ala, Ser, Thr, Pro, Asn, Gln.
7. according to the described antibacterial peptide of claim 3, it is characterized in that described basic aminoacids comprises Lys and Arg.
8. according to the described antibacterial peptide of claim 1, it is characterized in that antibacterial peptide has aminoacid sequence listed in the sequence table.
9. the purposes of the described cationic antibacterial peptide of claim 1-7 is characterized in that described antibacterial peptide is applied to prepare in the medicine for the treatment of gram-positive microorganism, Gram-negative bacteria and fungi infestation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200410020077 CN1597698A (en) | 2004-07-21 | 2004-07-21 | Cation antibacterial peptide |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200410020077 CN1597698A (en) | 2004-07-21 | 2004-07-21 | Cation antibacterial peptide |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1597698A true CN1597698A (en) | 2005-03-23 |
Family
ID=34663152
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 200410020077 Pending CN1597698A (en) | 2004-07-21 | 2004-07-21 | Cation antibacterial peptide |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1597698A (en) |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102229666A (en) * | 2011-05-27 | 2011-11-02 | 山东迅达康兽药有限公司 | Preparation method and application of animal-derived cationic antimicrobial peptides |
| CN102766196A (en) * | 2011-05-06 | 2012-11-07 | 上海医药工业研究院 | Cation antibacterial peptides, their preparation method and application |
| CN101696417B (en) * | 2009-11-02 | 2012-12-26 | 查向东 | Method for producing recombined cationic antibacterial peptide by adopting co-expression anionic ligand |
| CN103857702A (en) * | 2011-10-03 | 2014-06-11 | 南洋理工大学 | Cationic peptidopolysaccharides with excellent broad- spectrum antimicrobial activities and high selectivity |
| CN104356206A (en) * | 2011-05-06 | 2015-02-18 | 上海医药工业研究院 | HRP5 analogue and preparation method thereof |
| CN110684078A (en) * | 2019-10-21 | 2020-01-14 | 华中科技大学 | Cationic antibacterial peptide modified by dopamine or derivatives thereof, and preparation and application thereof |
| CN116178520A (en) * | 2023-03-09 | 2023-05-30 | 浙江大学医学院附属第一医院 | Cationic peptide and application thereof |
-
2004
- 2004-07-21 CN CN 200410020077 patent/CN1597698A/en active Pending
Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101696417B (en) * | 2009-11-02 | 2012-12-26 | 查向东 | Method for producing recombined cationic antibacterial peptide by adopting co-expression anionic ligand |
| CN102766196A (en) * | 2011-05-06 | 2012-11-07 | 上海医药工业研究院 | Cation antibacterial peptides, their preparation method and application |
| CN102766196B (en) * | 2011-05-06 | 2014-10-29 | 上海医药工业研究院 | Cation antibacterial peptides, their preparation method and application |
| CN104356206A (en) * | 2011-05-06 | 2015-02-18 | 上海医药工业研究院 | HRP5 analogue and preparation method thereof |
| CN104356206B (en) * | 2011-05-06 | 2017-07-14 | 上海医药工业研究院 | Hrp5 analogs and preparation method thereof |
| CN102229666A (en) * | 2011-05-27 | 2011-11-02 | 山东迅达康兽药有限公司 | Preparation method and application of animal-derived cationic antimicrobial peptides |
| CN103857702A (en) * | 2011-10-03 | 2014-06-11 | 南洋理工大学 | Cationic peptidopolysaccharides with excellent broad- spectrum antimicrobial activities and high selectivity |
| CN103857702B (en) * | 2011-10-03 | 2018-01-26 | 南洋理工大学 | Cationic peptide polysaccharide with excellent broad spectrum antimicrobial acivity and high selectivity |
| CN110684078A (en) * | 2019-10-21 | 2020-01-14 | 华中科技大学 | Cationic antibacterial peptide modified by dopamine or derivatives thereof, and preparation and application thereof |
| CN110684078B (en) * | 2019-10-21 | 2021-07-27 | 华中科技大学 | Cationic antibacterial peptide modified by dopamine or derivatives thereof, and preparation and application thereof |
| CN116178520A (en) * | 2023-03-09 | 2023-05-30 | 浙江大学医学院附属第一医院 | Cationic peptide and application thereof |
| CN116178520B (en) * | 2023-03-09 | 2024-04-19 | 浙江大学医学院附属第一医院 | Cationic peptide and application thereof |
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