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Problems solved by technology
Therefore, it is difficult to develop an antibacterial agent with antibacterial activity and antibacterial spectrum antibacterial performance that exceeds the antibacterial activity and antibacterial spectrum of inherent peptides in nature, as long as the peptide is based on the inherent antibacterial polypeptide.
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Embodiment 1
[0123] Embodiment 1: the chemical synthesis of antibacterial polypeptide
[0124] A total of 36 kinds of polypeptides (samples 1 to 33, reference products 1 to 3) were produced using the peptide synthesizer described below. Table 1 lists the amino acid sequences of these polypeptides.
[0125] Sample serial number
amino acid sequence
total number of amino acid residues
sample 1
PKKKRKV (serial number 4)
7
sample 2
RQARRNRRRRWR (serial number 21)
12
sample 3
RIRKKLR (serial number 43)
7
Sample 4
PPRKKRTVV (serial number 28)
9
Sample 5
RKKRRQRRR (serial number 20)
9
Sample 6
PRRRK (serial number 26)
5
Sample 7
RKKKRKV (serial number 83)
7
Sample 8
PKKKRKVLPPLERLTL (serial number 84)
16
Sample 9
LPPLERLTLPKKKRKV (serial number 85)
16
sample 10
RQARRNRRRRWRLPPPLERLTLD (serial number ...
Embodiment 2
[0151] Embodiment 2: the antibacterial activity of synthetic polypeptide
[0152] With 96 wells (well) microculture plate, obtain antibacterial property polypeptide (sample 1~33) of the present invention and the polypeptide of reference product 1~3 by liquid medium dilution method to Gram-negative bacteria (Escherichia coli: Escherichia coli: E. .coli) and Gram-positive bacteria (Staphylococcus aureus: S.aureus) antibacterial activity (minimum inhibitory concentration: MIC).
[0153] That is, liquid gravy medium (DIFCO product "NUTRIENTBROTH Dehydrated") with peptide concentrations of 500, 250, 125, 62.5, 31.3, 15.6, 7.8, 3.9, 1.9, 1.0, and 0.5 μM were prepared and filled into 96-well microculture plate. In addition, the bacterial liquid (approximately 2 × 10 6 cells / mL) and the drug solution (the above-mentioned peptide-containing broth medium) were inoculated in each well of a 96-well microplate. After inoculation, culture was started in a 37°C incubator, and sterility wa...
Embodiment 3
[0160] Embodiment 3: antibacterial polypeptide
[0161] Then, using various bacteria (see Table 3) as objects, the same antibacterial test (MIC measurement test) as in Example 2 was performed on the polypeptides of samples 16, 17, 10, 14 and 20, and the antibacterial spectrum of these polypeptides was evaluated. The results are shown in Table 3, and it was confirmed that the antibacterial polypeptide of the present invention has broad-spectrum antibacterial activity against both Gram-negative bacteria and Gram-positive bacteria.
[0162] Test bacteria
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Abstract
The present invention provides an artificially synthesized antimicrobial polypeptide that does not occur in nature and an antimicrobial agent comprising this polypeptide as the main component. This polypeptide contains in its peptide chain one or more types of nuclear localization signal sequences (NLSs) and / or one or more units of sequences prepared by partially modifying the NLS(s). The one or more units of NLSs and / or modified NLSs contain at least 5 amino acid residues in total and amount to 30% or more of the total amino acid residues constituting the peptide chain.
Description
technical field [0001] The present invention relates to an antibacterial polypeptide having an independent peptide chain structure that does not exist in nature, and an antibacterial agent (pharmaceutical composition) containing the polypeptide as a main component. Background technique [0002] It is reported that antibacterial peptides have a wide antibacterial spectrum, and it is difficult for drug-resistant bacteria to appear. Therefore, it is expected to be used for the prevention and treatment of bacterial infectious diseases in humans and animals, and the antibacterial peptides for the purpose of imparting antibacterial properties to food and other items. application. So far, many antibacterial polypeptides have been isolated from animals and plants. [0003] For example, Japanese Patent Application Laid-Open No. 2000-63400 discloses an antibacterial polypeptide derived from Taiwan beetle and an antibacterial agent containing the antibacterial polypeptide as an active...
Claims
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