Broken or folded helical peptide or peptide analog exhibiting antimicrobial activity against gram-negative bacteria, and use thereof

Patent metadata

Field	Value
Publication number	US-2018339016-A1
Application number	US-201615761089-A
Country	US
Kind code	A1
Filing date	Sep 19, 2016
Priority date	Sep 17, 2015
Publication date	Nov 29, 2018
Grant date	—

Abstract

The present invention relates to a broken or kinked helical peptide or peptide analogue and the use thereof, and more particularly to a Gram-negative bacterial membrane-penetrating peptide or peptide analogue wherein an alpha-helical amphipathic peptide composed of hydrophobic amino acids and hydrophilic amino acids has a kinked structure, or an antimicrobial composition employing the specific activity of the peptide against the Gram-negative bacterial membrane, or an antimicrobial composition for co-administration, or a conjugate comprising a drug linked to the peptide or peptide analogue, or an antibiotic comprising the same.

First claim

1 . A Gram-negative bacterial membrane-penetrating peptide or peptide analogue wherein an amphipathic alpha-helical peptide composed of hydrophobic amino acids and hydrophilic amino acids has a kinked structure, the peptide or peptide analogue comprising:
i) an alpha-helical structure wherein one or more hydrophobic amino acids of the amphipathic alpha-helical peptide or peptide analogue are kinked by substitution with one or more selected from the group consisting of proline (P), aspartic acid (D), asparagine (N), glutamic acid (E), glutamine (Q), D-form amino acids thereof, and derivatives thereof; or ii) an alpha-helical structure wherein two or more amino acids of the amphipathic alpha-helical peptide are kinked by linkage through a disulfide bond, a carbon-carbon bond, a maleimide bond or an amide bond at one or more positions selected from the group consisting of i, i+3, i+4, i+7, i+8, i+10 and i+11 (wherein i is an integer).

2 . The Gram-negative bacterial membrane-penetrating peptide or peptide analogue of claim 1 , wherein the peptide or peptide analogue exhibits any one of the following characteristics:
i) it has activity against Gram-negative bacteria without having hemolytic activity against host cells or activity against Gram-positive bacteria; ii) it is capable of binding to an LPS layer on the surface of a Gram-negative bacterial outer membrane; iii) it is capable of binding to an LPS layer on the surface of a Gram-negative bacterial outer membrane while entering the outer membrane and staying only in the outer membrane; iv) it has the property of penetrating a Gram-negative bacterial outer membrane and staying only in the outer membrane while having no ability to degrade the outer or inner membrane; v) it has a relatively strong hydrophilicity compared to a peptide having the same amino acid composition; vi) it has a structure wherein the alpha-helix is kinked by Pro, and the kinked portion is hydrophobic; and vii) the content of positively charged amino acids of 35% or more based on the total content of amino acids, or the content of hydrophobic amino acids is 35% or more based on the total content of amino acids.

3 . The Gram-negative bacterial membrane-penetrating peptide or peptide analogue of claim 1 , wherein the amphipathic alpha-helical peptide is composed of 12-20 amino acids comprising hydrophobic amino acids and hydrophilic amino acids.

4 . The Gram-negative bacterial membrane-penetrating peptide or peptide analogue of claim 1 , wherein the hydrophilic amino acids of the amphipathic alpha-helical peptide comprise one or more selected from the positively charged amino acid group consisting of arginine, lysine, and histidine.

5 . The Gram-negative bacterial membrane-penetrating peptide or peptide analogue of claim 1 , wherein the hydrophobic amino acids is one or more selected from the group consisting of leucine, valine, tryptophan, phenylalanine, tyrosine, isoleucine, D-form amino acids thereof, and derivatives thereof.

6 . The Gram-negative bacterial membrane-penetrating peptide or peptide analogue of claim 1 , wherein the amphipathic alpha-helical peptide comprises a amino-acid sequence represented by any one of the following formulas (1) to (8), the reverse sequence thereof, or a sequence containing the same repeatedly:
XXZYX  (1)
XYZYY  (1-1)
YXXZYXY  (2)
YXYZYYX  (2-1)
YYXXZYXYY  (3)
YYXYZYYXY  (3-1)
XYYXXZYXYYX  (4)
YYYXYZYYXYX  (4-1)
YXZZXX  (5)
YYXZZXXY  (6)
XYYXZZXXYY  (7)
XXYYXZZXXYYX  (8)
wherein X is a hydrophilic amino acid; Y is a hydrophobic amino acid; and Z is proline, aspartic acid, asparagine, glutamic acid, glutamine, or a derivative thereof which is an amino acid substituted for a kinked or broken structure.

7 . (canceled)

8 . (canceled)

9 . (canceled)

10 . (canceled)

11 . (canceled)

12 . (canceled)

13 . (canceled)

14 . The Gram-negative bacterial membrane-penetrating peptide or peptide analogue of claim 1 , wherein the amphipathic alpha-helical peptide comprises one or more amino acid residues, selected from the group consisting of positively charged arginine, lysine and histidine, in an amount equal to 35% or more of the total amino acids of the peptide, or
wherein the amphipathic alpha-helical peptide comprises one or more hydrophobic amino acid residues, selected from the group consisting of leucine, tryptophan, valine, phenylalanine, tyrosine, and isoleucine, in an amount equal to 35% or more of the total amino acids of the peptide.

15 . (canceled)

16 . The Gram-negative bacterial membrane-penetrating peptide or peptide analogue of claim 1 , wherein the amphipathic alpha-helical peptide comprises a sequence selected from the group consisting of the following SEQ ID NOs: 1 to 6, the reverse sequence thereof, or a sequence containing the same repeatedly:

(SEQ ID NO: 1)

KLLKL

(SEQ ID NO: 2)

LKKLL

(SEQ ID NO: 3)

LKKLLKL

(SEQ ID NO: 4)

KLLKLLK

(SEQ ID NO: 5)

LKKLLKLLKKLLKL

and

(SEQ ID NO: 6)

KLLKLLKKLLKLLK .

17 . (canceled)

18 . The Gram-negative bacterial membrane-penetrating peptide or peptide analogue of claim 1 , wherein the amino acids at one or more positions selected from the group consisting of positions 6, 7, 8, 9, 11 and 12 in the N-terminus to C-terminus direction in the amphipathic alpha-helical peptide are substituted.

19 . (canceled)

20 . (canceled)

21 . An antimicrobial composition for co-administration, comprising the peptide or peptide analogue of claim 1 .

22 . The composition of claim 21 , wherein the peptide is co-administered with one or more selected from the group consisting of hydrophobic compounds having a log P (partition coefficient) value of 0.19 or higher, compounds positively charged under physiological pH conditions, and colistin.

23 . A conjugate comprisi

Assignees

• Seoul Nat Univ R&Db Foundation

Inventors

• Yu Jaehoon
• Lee Yan
• Hyun Soonsil
• Kim Seo Yeon
• Jin Sun Mi
• Choi Yunhwa
• Jo Doyeon
• Lee Seonju

Classifications

• A61K38/04

  Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof ({enzyme inhibitors A61K38/005;} gastrins {A61K38/2207}, somatostatins A61K38/31, melanotropins A61K38/34 {; protease inhibitors A61K38/55}) · CPC title

• A61K31/431

  containing further heterocyclic rings, e.g. ticarcillin, azlocillin, oxacillin · CPC title

• A61K38/16 Primary

  Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof {(enzyme inhibitors A61K38/005)} · CPC title

• A61K47/6455

  Polycationic oligopeptides, polypeptides or polyamino acids, e.g. for complexing nucleic acids · CPC title

• A61K31/366

  having six-membered rings, e.g. delta-lactones · CPC title