EP0533795A1 - Composition et traitement utilisant des peptides biologiquement actifs contenant des resites d'aminoacides d - Google Patents

Composition et traitement utilisant des peptides biologiquement actifs contenant des resites d'aminoacides d

Info

Publication number
EP0533795A1
EP0533795A1 EP91911544A EP91911544A EP0533795A1 EP 0533795 A1 EP0533795 A1 EP 0533795A1 EP 91911544 A EP91911544 A EP 91911544A EP 91911544 A EP91911544 A EP 91911544A EP 0533795 A1 EP0533795 A1 EP 0533795A1
Authority
EP
European Patent Office
Prior art keywords
peptide
amino acid
basic
residue
amino acids
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.)
Withdrawn
Application number
EP91911544A
Other languages
German (de)
English (en)
Other versions
EP0533795A4 (en
Inventor
W. Lee Maloy
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Magainin Pharmaceuticals Inc
Original Assignee
Magainin Sciences Inc
Magainin Pharmaceuticals Inc
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Magainin Sciences Inc, Magainin Pharmaceuticals Inc filed Critical Magainin Sciences Inc
Publication of EP0533795A1 publication Critical patent/EP0533795A1/fr
Publication of EP0533795A4 publication Critical patent/EP0533795A4/en
Withdrawn legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • This invention relates to biologically active peptides, and more particularly to biologically active peptides wherein each amino acid residue of such peptides is a D-amino acid residue or glycine.
  • a compound comprising an analogue of Magainin I Peptide or Magainin II peptide.
  • Each amino acid residue of the Magainin I peptide and the Magainin II peptide is a D-amino acid residue or glycine.
  • the Magainin I or Magainin II peptide is in an amide or carboxy terminated form.
  • Magainin I is represented by the following structural formula using the single letter amino acid code and the numbers below each amino acid residues refer to the position of the residue in the peptide.
  • Magainin II is represented by the following structural formula using the single letter amino acid code and the numbers below each amino acid residue refer to the position of the residue in the peptide:
  • the Magainin I or Magainin II peptide is substituted in at least one of positions 1-23.
  • the substituents which may be employed in each of positions 1-23 are shown in the following table:
  • D-tryptophan residues may be protected with a formyl group or be unprotected.
  • the D-phenylalanine residues, whether such residue is present in its normal position, or employed as a substitution residue, may be a normal
  • the peptide is a Magainin I peptide, and at least one of amino acid residues 8, 10, 13, 16, 18 and 19 is substituted with a D-alanine residue. In another embodiment, the Magainin I peptide has D-alanine residue
  • the Magainin I peptide has D-alanine residue substitutions at four of amino acid residues 3, 8, 16, 19 and 23, and the remaining one of these residues is substituted with a D-lysine residue.
  • amino acid 21 of a Magainin I peptide is substituted with a D-proline residue.
  • At least one of amino acid residues 3, 7, 8, 10, 18-21, and 23 of Magainin I is selected from amino acid residues 3, 7, 8, 10, 18-21, and 23 of Magainin I.
  • amino acid residues 3, 8, 9, 19 and 23 of Magainin I are each substituted with a D-lysine residue and amino acid residue 16 is substituted with a D-alanine residue.
  • Magainin I is substituted with a protected D-tryptophan residue, wherein the protecting group is preferably a formyl group.
  • the protecting group is preferably a formyl group.
  • at least one of amino acid residues 9, 13, 15 and 17 of Magainin I is substituted with an unprotected D-tryptophan residue.
  • the peptide is a Magainin II peptide, and at least one of amino acid residues 1-8, 10, 13, 14, 16 and 18-23 is a D-alanine residue.
  • At least one of amino acid residues 1-3, 5-9, 12, 13 and 15-23 of Magainin II is a D-lysine residue.
  • an analogue of Magainin I or Magainin II peptide said Magainin I or Magainin II peptide being in an amide- or carboxy-terminated form and having the structural formulas hereinabove described, wherein each amino acid residue of said Magainin I or Magainin II peptide is a D-amino acid residue or glycine, and wherein at least one of amino acid residues 15-23 is omitted and at least one of the remaining amino acid residues is substituted.
  • the substituents which may be employed in at least one of positions 1-23 are shown in the following table:
  • amino acid residues 16-23 of Magainin I are deleted, and amino acid residues 3, 8 and 10 are substituted with a D-alanine residue.
  • amino acid residue 19 of Magainin I is deleted, and preferably amino acid residues 5, 8, 9 and 16 are each substituted with a D-lysine residue, amino acid residue 21 is substituted with a D-leucine residue, and amino acid residues 18 and 23 are substituted with a D-alanine residue.
  • amino acid residues 17-23 or 16-23 or 15-23 are deleted, and amino acid residues 3, 7, and 8 are each
  • Preferred peptides are as follows:
  • amino acid residue 21 of Magainin I may be deleted, and preferably amino acid residues 5,10, 18, and 19 are each substituted with a D-lysine residue, amino acid residue 7 is substituted with a D-phenylalanine residue, and amino acid residue 22 is substituted with a D-alanine residue.
  • amino acid residue 19 is omitted, and at least one of amino acid residues 3, 7, 8, 10, 13, 15, 16, 18 21, 22 or 23 is substituted with another amino acid as follows:
  • the peptide is a Magainin II peptide
  • the substitution analogue wherein amino acid 19 is deleted is selected from the class consisting of the following substitution analogues:
  • K** is E-F-moc-lysine.
  • amide-terminated preferably Magainin II, wherein a portion of the basic peptide is deleted and at least one of the remaining amino acid residues is substituted as hereinabove described; in particular, amino acid residue 19 is omitted and in addition either amino acid residues 1-4 or 3,5, and 6 are omitted.
  • Preferred peptides are as follows:
  • a compound comprising a peptide deletion analogue of an amide or carboxy terminated Magainin I, wherein Magainin I is represented by the following structural formula using the single letter amino acid code and the numbers below each amino acid residue refer to the position of the residue in the peptide:
  • each amino acid residue is a D-amino acid residue or a glycine residue, and at least one of amino acid residues 15 through 23 is omitted.
  • at least one of amino acid residues 15, 16, 18, 19, 21, 22 and 23 is omitted.
  • at least amino acid residue 18 is omitted whereas in another embodiment, at least amino acid residue 19 is omitted, and in yet another embodiment at least amino acid residue 21 is omitted. In preferred embodiments, only one of amino acid residues 18, 19, and 21, respectively, is omitted.
  • amino acid residues 21, 22, and 23 are omitted, amino acid residues 19 through 23 are omitted, amino acid residues 18 through 23 are omitted, and amino acid residues 17 through 23 are omitted.
  • the compound can be a deletion analogue of amide - or carboxy-terminated Magainin I.
  • a compound comprising a peptide that is a deletion analogue of an amide - or carboxy-terminated Magainin II, wherein Magainin II is represented by the following
  • each amino acid residue is a D-amino acid residue or glycine, and at least one of amino acids 15 through 22 is omitted.
  • at least one of amino acids 15, 18, 19, 20, 21, and 22 is omitted.
  • at least amino acid 18 is omitted.
  • at least amino acid 19 is omitted.
  • Another embodiment omits at least amino acid 21, and yet other embodiment omits at least amino acid 22.
  • the compound can be a deletion analogue of amide-terminated Magainin II.
  • a basic (positively charged) polypeptide having at least sixteen amino acids wherein the polypeptide includes at least eight hydrophobic amino acids and at least eight hydrophilic amino acids.
  • Each of the amino acid residues of the polypeptide is a D-amino acid residue or a glycine residue.
  • the hydrophobic amino acids are in groups of two adjacent amino acids wherein the amino acids are D-amino acids or glycine, and each group of two
  • hydrophobic amino acids is spaced from another group of two hydrophobic amino acids by at least one D-amino acid other than a hydrophobic amino acid (preferably at least two D-amino acids) and generally by no greater than four D-amino acids, and the D-amino acids between pairs of hydrophobic amino acids may or may not be hydrophilic.
  • the hydrophilic amino acids are generally also in groups of two adjacent D-amino acids in which at least one of the two
  • D-amino acids is a basic hydrophilic amino acid, with such groups of two hydrophilic D-amino acids being spaced from each other by at least one amino acid, other than a hydrophilic D-amino acid, wherein each of said at least one amino acid(s) is a D-amino acid or glycine (preferably at least two amino acids) and generally no greater than four amino acids, and the amino acids between pairs of hydrophilic D-amino acids may or may not be hydrophobic.
  • the polypeptide comprises a chain of at least four groups of amino acids wherein each amino acid is a D-amino acid or glycine, with each group consisting of four amino acids wherein each of the at least four amino acids is a D-amino acid or glycine.
  • Two of the four amino acids in each group are hydrophobic amino acids, and two of the four amino acids in each group are hydrophilic, with at least one of the hydrophilic amino acids in each group being a basic hydrophilic amino acid and the other being a basic or neutral hydrophilic amino acid.
  • the hydrophobic amino acids may be selected from the class consisting of D-Ala, D-Cys, D-Phe, D-Ile, D-Leu, D-Met, D-Val, D-Trp, D-Tyr, and Gly.
  • the neutral hydrophilic D-amino acids may be selected from the class consisting of D-Ser, D-Asn, D-Gln, and D-Thr.
  • the basic hydrophilic D-amino acids may be selected from the class consisting of D-Lys, D-Arg, and D-His, and D-ornithine.
  • Each of the groups of four amino acids may be of the sequence ABCD, BCDA, CDAB, or DABC, wherein A and B are each hydrophobic amino acids and may be the same or different, one of C or D is a basic hydrophilic amino acid, and the other of C or D is a basic or neutral hydrophilic amino acid and may be the same or different .
  • the polypeptide chain may comprise 5 or 6 groups of this sequence. In each group, each of A, B, C and D may be the same in some or all of the groups or may be different in some or all of the groups.
  • the polypeptide chain preferably has at least 20 amino acids, and no greater than 50 amino acids wherein each amino acid is a D-amino acid or glycine. It is to be understood, however, that the polypeptide does not have to consist entirely of the groups described above.
  • the polypeptide may have amino acids extending from either or both ends of the noted groups forming the polypeptide chain and/or there may be amino acids between one or more of the at least four groups and still remain within the scope of the invention, provided that each amino acid residue of the polypeptide chain is a D-amino acid residue or glycine.
  • the groups of amino acids may be repeating groups of amino acids, or the amino acids in the various groups may vary provided that in each group of the at least four groups of amino acids there are two hydrophobic amino acids wherein each amino acid is a D-amino acid or glycine, and two hydrophilic D-amino acids as hereinabove noted.
  • the biologically active polypeptide comprises a chain including at least four groups of amino acids, each containing four amino acids wherein each amino acid is a D-amino acid or glycine. Two of the four amino acids in each group are hydrophobic, wherein each hydrophobic amino acid is a D-amino acid or glycine, at least one amino acid is a basic hydrophilic D-amino acid, and the remaining one is a basic or neutral hydrophilic D-amino acid, with the polypeptide chain preferably having at least 20 amino acids but no greater than 50 amino acids.
  • each of the at least four groups of amino acids which are in the peptide chain is of the sequence A-B-C-D, B-C-D-A, C-D-A-B or D-A-B-C wherein A and B are
  • polypeptide chain therefore, may have one of the following sequences:
  • X 2 is A-, D-A- or C-D-A- Y 2 is -B, -B-C or B-C-D X 3 is B-, A-B-, D-A-B-
  • Y 3 is -C, -C-D, -C-D-A
  • X 4 is C-, B-C-, A-B-C-
  • Y 4 is -D, -D-A, -D-A-B
  • n is at least 4.
  • the peptide may have amino acids extending from either end of the chain.
  • the chains may have a D-Ser-D-Lys sequence before the "D-Ala” end, and/or a D-Ala-D-Phe sequence after the "D-Lys" end.
  • Other amino acid sequences may also be attached to the "D-Ala” and/or the "D-Lys” end.
  • the chain may have, for example, a C-D sequence before the first A-B-C-D group.
  • other acid sequences including D-amino acids or glycine may be attached to the "A" and/or the "D" end of one of these polypeptide chains.
  • D-amino acids or glycine residues in the chain which space one or more groups of the hereinabove noted four amino acids from each other.
  • polypeptides are generally water-soluble to a
  • polypeptides are non-hemolytic, i.e., it will not rupture red blood cells at effective antimicrobial concentrations.
  • the structure of such polypeptides provides for flexibility of the polypeptide molecule. When the polypeptide is placed in water, it does not assume an amphiphilic structure.
  • polypeptide encounters an oily surface or membrane
  • polypeptide chain folds upon itself into a rod like structure.
  • peptide having from eight to fifteen amino acids comprised of at least four
  • hydrophobic amino acids and four hydrophilic amino acids Each amino acid residue is a D-amino acid residue or glycine.
  • the hydrophobic amino acids are in groups of two adjacent hydrophobic D-amino acids or glycine wherein each group of two hydrophobic D-amino acids or glycine is spaced from each other by at least one D-amino acid other than a hydrophobic D-amino acid or glycine (preferably at least two D-amino acids) and generally no greater than four D-amino acids, and the D-amino acid(s) between pairs of hydrophobic amino acids wherein each amino acid is a D-amino acid or glycine, may or may not be hydrophilic.
  • D-amino acids are generally also in groups of two adjacent amino acids
  • D-amino acids in which at least one of the two D-amino acids is a basic hydrophilic D-amino acid and the other of the two is basic or neutral.
  • the groups of two D-hydrophilic amino acids are spaced from each other by at least one amino acid other than a hydrophilic D-amino acid, and wherein the at least one amino acid is a D-amino acid or glycine (preferably at least two amino acids) and generally no greater than 4 amino acids, and the amino acids between pairs of hydrophilic amino acids may or may not be hydrophobic.
  • the peptide having from 8 to 15 amino acids is amphiphilic and is positively charged (basic).
  • the 8 to 15 amino acid peptide hereinabove described may or may not be bioactive, and in the case where such peptide is not bioactive, it has utility as an intermediate in providing the hereinabove noted peptides which have at least 16 amino acids and which are bioactive.
  • two peptides, one having eight D-amino acids and the other having twelve D-amino acids may be coupled to each other to form a peptide having 20 D-amino acids of the type hereinabove described and which is bioactive.
  • the peptides may be coupled by standard peptide chemistry techniques.
  • such peptides may be condensed in solution by the technique disclosed by Johnson, et al. Peptides, pages 239-42 (Walter de Gruzter & Co., 1986).
  • peptides represented by the following structure wherein A, B, C and D are as defined previously;
  • W 2 is A-, D-A-, C-D-A- W 3 is B-, A-B-, D-A-B- W 4 is C-, B-C-, A-B-C- Z 1 is -A, -A-B, -A-B-C Z 2 is -B, -B-C, -B-C-D
  • Z 3 is -C, -C-D, -C-D-A
  • Z 4 is -D, -D-A, -D-A-B
  • n 2 or 3
  • a is o or 1
  • b is o or 1.
  • peptide which includes the following basic peptide structure X:
  • R 1 is a hydrophobic amino acid
  • R 2 is a hydrophobic amino acid or a basic hydrophilic amino acid
  • R 3 is a basic hydrophilic amino acid
  • R 4 is a hydrophobic or neutral hydrophilic amino acid
  • R 5 is a basic or neutral hydrophilic amino acid, and each amino acid residue of said peptide is a D-amino acid residue or glycine.
  • the CPF peptides of the present invention may include only the hereinabove noted D-amino acids or glycine or may include additional D-amino acids or glycine residues at the amino and/or carboxyl end or both the amino and carboxyl end.
  • the peptide does not include more than 40 amino acids, wherein each amino acid is a D-amino acid or glycine.
  • the CPF peptides including the above basic structure preferably have from 1 to 4 additional amino acids at the amino end, wherein each amino acid is a D-amino acid or glycine.
  • X is the hereinabove described basic peptide structure and Y is (i) R 5 -, or
  • R 1 , R 2 and R 5 are as previously defined.
  • the carboxyl end of the basic peptide structure may also have additional amino acids which may range from 1 to 13
  • the basic structure may have from 1 to 7 additional amino acids at the carboxyl end, which may be represented as follows:
  • X is the hereinabove defined basic peptide structure and Z is
  • R 1 -R 1 -R 4 -R 4 -R 6 -D-Gln-D-Gln wherein R 1 and R 4 are as previously defined, and R 6 is D-proline or a hydrophobic amino acid.
  • Preferred peptides may be represented by the following structural formula
  • CPF peptides As representative examples of CPF peptides used in the present invention, there may be mentioned peptides represented by the following (single letter amino acid code):
  • Preferred CPF peptides are of the following sequences:
  • composition comprising at least one
  • biologically active peptide which includes at least a chain of the following 20 amino acid residues having the following peptide structure X 10 where X 10 is:
  • R 11 is a hydrophobic amino acid
  • R 12 is a basic hydrophilic amino acid
  • R 13 is a hydrophobic or basic hydrophilic amino acid
  • R 14 is a netural hydrophilic amino acid, or a basic
  • R 15 is a neutral hydrophilic amino acid
  • R 16 is D-proline or a hydrophobic amino acid
  • R 17 is a netural hydrophilic, basic hydrophilic or hydrophobic amino acid.
  • Each of the amino acid residues of the peptide is a D-amino acid residue or
  • the at least one biologically active peptide which includes peptide residue X 10 has at least the hereinabove
  • Such peptides are derived from fragments of the human hormone
  • cholecystokinin or may be derivatives of such fragments.
  • the at least one biologically active peptide is of the formula R 11 -R 11 -X-,
  • R 11 is a hydrophobic amino acid as described above.
  • the biologically active peptide of this embodiment includes a chain of at least the following amino acids:
  • the at least one biologically active petpide includes a chain of at least the following amino acids:
  • R 11 , R 12 , and R 15 are amino acids of the types hereinabove described.
  • the peptide may be amide-terminated or carboxy-terminated.
  • a biologically active amphiphilic peptide which includes the following basic structure X 12 :
  • R 21 is a basic hydrophilic amino acid
  • R 22 is a hydrophobic amino acid
  • R 23 is a neutral hydrophilic or hydrophobic amino acid
  • n is from 2 to 5
  • each amino acid residue of the peptide is a D-amino acid residue or glycine.
  • the peptide may include the following structure:
  • the peptide may include the following structure:
  • the peptide may include the following structure:
  • n 3
  • the peptide is of the following structure as indicated by the single letter amino acid code:
  • n 2
  • the peptide preferably is of the following structure as indicated by the single letter amino acid code:
  • a biologically active amphiphilic peptide which includes the following basic structure X 14 :
  • the peptide may include the following structure: Y 14 -X 14 , wherein X 14 is as hereinabove described, and Y14 is:
  • the peptide may include the following structure:
  • the peptide is of the following structural formula as indicated by the single letter amino acid code:
  • the peptide is of the following structural formula as indicated by the single letter amino acid code:
  • the peptide may include the following basic peptide structure X 16 :
  • R 31 is a hydrophobic amino acid
  • R 32 is a basic
  • R 33 is a neutral hydrophilic, basic hydrophilic, or hydrophobic amino acid
  • R 34 is a hydrophobic or basic hydrophilic amino acid
  • each amino acid residue is a D-amino acid residue or glycine.
  • R 33 is a neutral hydrophilic amino acid.
  • Such peptides are commonly referred to as PGLa peptides.
  • the PGLa peptides generally include at least seventeen amino acids and may include as many as forty amino acids.
  • the hereinabove described basic peptide structure for a PGLa peptide may include additional amino acids at the amino end or at the carboxyl end or at both the amino and
  • a PGLa peptide may have the following structure:
  • R 31 and R 34 are as previously defined.
  • a PGLa peptide may also have the following structure:
  • R 31 is as previously defined.
  • a PGLa peptide may also have the following structure: (Y 16 ,) a -X 16 -(Z 16 )b
  • X 16 , Y 16 , and Z 16 are as Previously defined, a is 0 or 1 and b is 0 or 1.
  • the peptide may include the following basic peptide structure X 18 :
  • R 31 -R 31 -R 31 -R 32 -(R 36 ) n -R 31 -- wherein R 31 , R 32 , R 33 , and R 34 are as previously defined, and R 35 is glutamine or asparagine or a basic hydrophilic, or hydrophobic amino acid, R 36 is glutamic acid, aspartic acid, a hydrophobic amino acid or a basic hydrophilic amino acid, and n is 0 or 1.
  • Each amino acid residue is a D-amino acid residue or glycine.
  • Such peptides are commonly referred to as XPF peptides.
  • the XPF peptides generally include at least nineteen amino acids and may include up to forty amino acids. Accordingly, the hereinabove described basic peptide structure of XPF may include additional amino acids at the amino and carboxyl ends.
  • an XPF peptide may include the following structure:
  • R 31 and R 34 are as previously defined.
  • An XPF peptide may include the following structure:
  • An XPF peptide may also have the following structure
  • XPF or PGLa peptides which are characterized by the following primary amino acid sequence (single letter amino acid code):
  • PGLa GMASKAGAIAGKIAKVALKAL (NH 2 )
  • the above-mentioned peptides also have increased resistance to proteolytic enzymes while retaining their biological activity.
  • the peptides hereinabove described, and/or analogues or derivatives thereof are generally water soluble to a concentration of at least 20 mg/ml at neutral pH in water.
  • such peptides are non-hemolytic; i.e., they will not rupture blood cells at effective concentrations.
  • the peptides cause less than 5% hemolysis of human erythrocytes at a
  • the structure of such peptide provides for flexibility of the peptide molecule. When the peptide is placed in water, it does not assume an amphiphilic structure. When the peptide encounters an oily surface or membrane, the peptide chain folds upon itself into a rod-like structure.
  • the peptides may be C-terminal acids "or amides.
  • the peptides may be administered to a host; for example a human or non-human animal, in an amount effective to inhibit growth of a target cell or virus.
  • a host for example a human or non-human animal, in an amount effective to inhibit growth of a target cell or virus.
  • peptides and/or analogues or derivatives thereof may be used as antimicrobial agents, anti-viral agents, antibiotics, anti-tumor agents, antiparasitic agents, spermicides, as well as exhibiting other bioactive functions.
  • antimicrobial means that the peptides of the present invention inhibit, prevent, or destroy the growth or proliferation of microbes such as bacteria, fungi, or the like.
  • peptides employed in the present invention produce effects adverse to the normal biological functions of the cell, tissue, or organism including death or destruction and prevention of the growth or proliferation of the biological system when contacted with the peptides.
  • spermicidal as used herein means that the peptides employed in the present invention, inhibit, prevent, or destroy the motility of sperm.
  • antiviral means that the peptides employed in the present invention inhibit, prevent, or destroy the growth or proliferation of viruses.
  • anti-tumor means that the peptide inhibits the growth of or destroys tumors.
  • antiparasitic means that the peptides of the present invention may be used to inhibit the growth of or destroy parasites.
  • the peptides of the present invention have a broad range of potent antibiotic activity against a plurality of microorganisms including Gram-positive and Gram-negative bacteria, fungi, protozoa, and the like, as well as parasites.
  • the peptides of the present invention allow a method for treating or controlling microbial infection caused by organisms which are sensitive to the peptides. Such treatment may comprise administering to a host organism or tissue susceptible to or affiliated with a microbial infection an antimicrobial amount of at least one of the peptides. Because of the antibiotic properties of the peptides, they may also be used as preservatives or sterilants of materials susceptible to microbial contamination.
  • the peptide may be administered in combination with a non-toxic pharmaceutical carrier or vehicle such as a filler, non-toxic buffer, or physiological saline solution.
  • a non-toxic pharmaceutical carrier or vehicle such as a filler, non-toxic buffer, or physiological saline solution.
  • compositions may be used topically or systemically and may be in any suitable form such as a liquid, solid,
  • compositions may also be used in combination with adjuvants, protease inhibitors, or compatible drugs where such a combination is seen to be desirable or
  • microorganisms including protozoa viruses, and the like, as well as by parasites.
  • the peptides of the present invention may be administered to a host; in particular an animal, in an effective antibiotic and/or anti-tumor and/or anti-viral and/or anti-microbial and/or anti-parasitic and/or an antispermicidal amount.
  • composition in accordance with the invention will contain an effective anti-microbial amount and/or an effective antispermicidal amount and/or an effective
  • the peptide of the present invention may also be employed in promoting or stimulating healing of a wound in a host.
  • wound healing includes various aspects of the would healing process.
  • peptides increase wound breaking strength.
  • the peptides of the present invention may also be employed so as to reverse the inhibition of wound healing caused by steroids such as cortisone.
  • the peptides of the present invention may be used in the treatment of external burns and to treat and/or prevent skin and burn infections.
  • the peptides may be used to treat skin and burn infections caused by organisms such as, but not limited to, P. aeruginosa and S . aureus.
  • the peptides are also useful in the prevention or treatment of eye infections.
  • infections may be caused by bacteria such as, but not limited to, P. aeruginosa, S . aureus, and N. gonorrhoeae, by fungi such as but not limited to C. albicans and A. fumigatus, by parasites such as but not limited to A.
  • the peptides may also be effective in killing cysts, spores, or trophozoites of infection - causing organisms.
  • Such organisms include, but are not limited to Acanthamoeba which forms trophozoites or cysts, C. albicans, which forms spores, and A. fumigatus, which forms spores as well.
  • the peptide is employed to provide peptide dosages of from 0.1 mg. to 500 mg. per kilogram of host weight.
  • the peptide is used in a
  • the peptides may be produced by known techniques and obtained in substantially pure form.
  • the peptides may be synthesized on an automatic synthesizer. Journal of the American Chemical Society, Vol. 85, pgs. 2149-54 (1963). It is also possible to produce such peptides by genetic engineering techniques.
  • the peptides of the present invention may be administered alone, or in combination with other such peptides.
  • the peptides may also be administered in combination with biologically active peptides or proteins containing L-amino acid residues, such as ion channel-forming proteins, or the peptides of the present invention may be administered in combination with other active components, such as toxic ions or antibiotics.
  • Biologically active peptides containing L-amino acid residues which may be administered in combination with the peptides of the present invention include magainin peptides, PGLa peptides, XPF peptides, cecropins, and sarcotoxins.
  • Magainin peptides are described in Proc. Natl. Acad. Sci., Vol. 84, pgs 5449-53 (Aug. 1987).
  • a review of XPF and PGLa peptides may be found in Hoffman, et al., EMBOJ., 2:711-714 (1983). Andreu, et al., J. Biochem, 149:531-535 (1985); Gibson, et al., Biochem J., 243:113-120 (1987).
  • cecropins and analogues and derivatives thereof are described in Ann. Rev. Microbiol., Vol. 41, pgs, 103-26 (1987), and Christensen, et al., PNAS, Vol. 85, pgs. 5072-5076. (1988).
  • Sarcotoxins and analogues and derivatives thereof are described in Molecular Entomotogy, pgs 369-78, Alan R. Liss, Inc. (1987).
  • Ion channel-forming proteins or peptides which may be employed in combination with the peptides of the present
  • defensins also known as human neutrophil antimicrobial peptides (HNP), major basic protein (MBP) of eosinophils, bactericidal permeability-increasing protein (BPI), and a pore-forming cytotoxin called variously perforin,
  • cytolysin or pore-forming protein.
  • Defensins are described in Selsted, et al., J. Clin. Invest., Vol. 76, pgs. 1436-1439
  • MBP proteins are described in Wasmoen, et al., J. Biol. Chem., Vol. 263, pgs. 12559-12563 (1988).
  • BPI proteins are described in Ooi, et al., J. Biol. Chem. Vol. 262, pgs.
  • Toxic ions which may be employed in combination with the peptides of the present invention include anions such as fluoride, peroxide, and bicarbonate anions, and cations such as silver, zinc, mercury, arsenic, copper, platinum, antimony, gold, thallium, nickel, selenium, bismuth, and cadmium cations.
  • a toxic ion is one which when introduced into a target cell inhibits and/or prevents and/or destroys the growth of the target cell.
  • Such a toxic ion is one which in the absence of an ion channel forming peptide is unable to cross a natural or synthetic lipid membrane; in particular a cell membrane, in sufficient amounts to affect a cell adversely.
  • the peptide and the toxic ion are employed in amounts effective to inhibit and/or prevent and/or destroy the growth of the target cell.
  • the ion is administered or prepared in a single composition or in separate compositions, employed in amounts effective to inhibit and/or prevent and/or destroy the growth of the target cell.
  • the amount of toxic ion is effective to reduce the maximum effective concentration of the peptide or protein for inhibiting growth of a target cell.
  • Antibiotics which may be employed in combination with the peptides of the present invention include bacitracins,
  • hydrophobic antibiotics including macrolide antibiotics, penicillins, m ⁇ nobactams, or derivatives or
  • antibiotics which are 50-S ribosome inhibitors such as lincomycin; clindamycin; and chloramphenicol; etc.; and antibiotics which have a large lipid like lactone ring, such as mystatin; pimaricin, etc.
  • the peptide and antibiotic may be adminstered by direct administration to a target cell or by systemic or topical administration to a host which includes the target cell, in order to prevent, destroy or inhibit the growth of a target cell.
  • Target cells whose growth may be prevented, inhibited, or destroyed by the administration of the peptides and antibiotic include Gram-positive and Gram-negative bacteria as well as fungal cells.
  • each amino acid residue is a D-amino acid residue or glycine, was prepared at a concentration of 512 ⁇ g/ml in sterile deionized distilled water and stored at -70°C.
  • the stock peptide solution is diluted in serial dilutions (1:2) down the wells of a microtiter plate so that the final concentrations of peptides in the wells are 0.25, 0.50, 1, 2, 4, 8, 16, 32, 64, 128, and 256 ⁇ g/ml.
  • 1.5 X 10 5 CFUs/ml of either S. aureus ATCC 25923, E. coli ATCC 25922, or P. aeruginosa ATCC 27853 were added to the wells in full strength Mueller Hinton broth (BBL 11443) from a mid-log culture.
  • the inoculum is standarized spectrophotometrically at 600nm and is vertified by colony counts.
  • the plates are incubated for 16-20 hours at 37°C, and the minimal inhibitory concentration (MIC) for each peptide is determined.
  • Minimal inhibitory concentration is defined as the lowest concentration of peptide which produces a clear well in the microtiter plate.
  • the minimal inhibitory concentration of the peptide was 8 ⁇ g/ml against S. aureus, 32 ⁇ g/ml against P. aeruginosa, and 4 ⁇ g/ml against E. coli.
  • a stock solution of the above-mentioned peptide was prepared as hereinabove described wherein each amino acid residue was not a D-amino acid residue. This peptide was then tested as hereinabove described for MIC against S. aureus, P. aeruginosa, and E. coli.
  • the MIC of the peptide was 8 ⁇ g/ml against S. aureus, 16 ⁇ g/ml against P.
  • peptides of the present invention whether administered alone or in combination with agents such as toxic ions,
  • antibiotics or other biologically active peptides or proteins as hereinabove described, may be employed in a wide variety of pharmaceutical compositions in combination with a non-toxic pharmaceutical carrier or vehicle such as a filler, non-toxic buffer, or physiological saline solution.
  • a non-toxic pharmaceutical carrier or vehicle such as a filler, non-toxic buffer, or physiological saline solution.
  • Such pharmaceutical compositions may be used topically or systemically and may be in any suitable form such as a liquid, solid, semi-solid, injectable solution, tablet, ointment, lotion, paste, capsule or the like.
  • the peptide and/or agent as hereinabove described may also be used in combination with adjuvants, protease inhibitors, or compatible drugs where such a combination is seen to be desirable or advantageous in controlling infection caused by harmful microorganisms including protozoa, viruses, and the like.
  • the peptide may be administerd to a host, in particular an animal, in an effective antibiotic and/or anti-tumor and/or antiviral and/or antimicrobial and/or antispermicidal and/or antiparasitic amount, or in an amount effective to stimulate wound healing in a host.
  • the peptides may be administerd either alone or in combination with a toxic ion, antibiotic, or ion channel forming peptide or protein as hereinabove described.
  • the activity of the peptide is potentiated.
  • the peptide When the peptide is administered in combination with an agent as hereinabove described, it is possible to administer the peptide and agent in separate forms.
  • the agent may be administered systemically and the peptide may be administered topically.
  • the peptide When the peptide is adminitered topically, it may be administered in combination with a water-soluble vehicle, said water-soluble vehicle being in the form of an ointment, cream, lotion, paste or the like.
  • water-soluble vehicles which may be employed include, but are not limited to, glycols, such as polyethylene glycol, hydroxycellulose, and KY Jelly.
  • the water-soluble vehicle is preferably free of an oily substance.
  • the peptide may also be employed alone, or in combination with other peptides of the present invention, or in combination with peptides containing L-amino acid residues, or in combination with other active components (eg., toxic ions, antibiotics) as hereinabove described in the form of an oral composition for oral hygiene.
  • Such a composition may be incorporated into a wide variety of compositions and materials used for oral hygiene purposes, which include, but are not limited to, toothpastes, mouthwashes, tooth gels, and tooth powders.
  • Such composition may thus be used to treat or prevent periodontal disease, to prevent or reduce plaque, and/or to prevent or treat or reduce dental caries.
  • the peptide may be used to inhibit, prevent, or destroy the growth of Streptococcus mutans, which is associated with dental caries and periodontal disease.

Abstract

L'invention se rapporte à un peptide biologiquement actif dont chaque reste d'aminoacide est un reste d'aminoacide D ou un reste de glycine. De tels peptides, dans lesquels chaque reste d'aminoacide est un reste d'aminoacide D ou un reste de glycine, sont par exemple des analogues de suppression et de substitution de peptides de magaïnine, des peptides CPF, des peptides PGLa, un peptide XPF et des dérivés de cholécystokinine. Ces peptides possèdent une résistance accrue aux enzymes protéolytiques, tout en conservant leur activité biologique.
EP19910911544 1990-05-14 1991-05-09 Composition of and treatment with biologically active peptides having d-amino acid residues Withdrawn EP0533795A4 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US52268890A 1990-05-14 1990-05-14
US522688 1990-05-14

Publications (2)

Publication Number Publication Date
EP0533795A1 true EP0533795A1 (fr) 1993-03-31
EP0533795A4 EP0533795A4 (en) 1993-10-20

Family

ID=24081910

Family Applications (1)

Application Number Title Priority Date Filing Date
EP19910911544 Withdrawn EP0533795A4 (en) 1990-05-14 1991-05-09 Composition of and treatment with biologically active peptides having d-amino acid residues

Country Status (4)

Country Link
EP (1) EP0533795A4 (fr)
JP (1) JPH05507273A (fr)
CA (1) CA2042468A1 (fr)
WO (1) WO1991017760A1 (fr)

Families Citing this family (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6348445B1 (en) 1992-06-01 2002-02-19 Magainin Pharmaceuticals, Inc. Biologically active peptides with reduced toxicity in animals and a method for preparing same
US5654274A (en) * 1992-06-01 1997-08-05 Magainin Pharmaceuticals, Inc. Biologically active peptides having N-terminal substitutions
US5593866A (en) * 1992-08-21 1997-01-14 The University Of British Columbia Cationic peptides and method for production
US5733872A (en) * 1993-03-12 1998-03-31 Xoma Corporation Biologically active peptides from functional domains of bactericidal/permeability-increasing protein and uses thereof
US6057291A (en) 1995-06-02 2000-05-02 University Of British Columbia Antimicrobial cationic peptides
JP3860838B2 (ja) 1995-08-23 2006-12-20 ユニバーシティー オブ ブリティッシュ コロンビア 抗微生物性陽イオン性ペプチドおよびそのスクリーニング方法

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1988006597A1 (fr) * 1987-03-04 1988-09-07 The United States Of America, As Represented By Th Composes bioactifs synthetiques nouveaux et procede d'obtention d'effets bioactifs
WO1991012015A1 (fr) * 1990-02-08 1991-08-22 Magainin Sciences Inc. Peptides amphiphiles biologiquement actifs et procede pour inhiber la croissance de cellules cibles, de virus ou de cellules contaminees par virus

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4659692A (en) * 1982-11-19 1987-04-21 The Regents Of The University Of California Cationic oligopeptides having microbicidal activity
DE3324534A1 (de) * 1983-07-07 1985-01-17 Ciba-Geigy Ag, Basel Modifizierte protease-inhibitoren, verfahren zu ihrer herstellung und daraus bereitete pharmazeutische mittel
US4617149A (en) * 1983-09-21 1986-10-14 Eli Lilly And Company Growth hormone release factor analogs
DE3438296A1 (de) * 1984-04-18 1985-11-07 Hoechst Ag, 6230 Frankfurt Neue polypeptide mit blutgerinnungshemmender wirkung, verfahren zu deren herstellung bzw. gewinnung, deren verwendung und diese enthaltende mittel
EP0209061B1 (fr) * 1985-07-17 1994-01-12 Hoechst Aktiengesellschaft Peptides à activité anticoagulante, leur procédé de préparation, d'obtention, leur application et les agents les contenant

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1988006597A1 (fr) * 1987-03-04 1988-09-07 The United States Of America, As Represented By Th Composes bioactifs synthetiques nouveaux et procede d'obtention d'effets bioactifs
WO1991012015A1 (fr) * 1990-02-08 1991-08-22 Magainin Sciences Inc. Peptides amphiphiles biologiquement actifs et procede pour inhiber la croissance de cellules cibles, de virus ou de cellules contaminees par virus

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
FEBS LETTERS vol. 274, no. 1,2, November 1990, pages 151 - 155 R. BESALLE ET AL. 'All-D-mangainin: chirality, antimicrobial activity and proteolytic resistance' *
PROC. NATL. ACAD. SCI. USA vol. 87, June 1990, pages 4761 - 4765 D.WADE ET AL. 'All-D amino acid-containing channel-forming antibiotic peptides' *
See also references of WO9117760A1 *

Also Published As

Publication number Publication date
EP0533795A4 (en) 1993-10-20
WO1991017760A1 (fr) 1991-11-28
CA2042468A1 (fr) 1991-11-15
JPH05507273A (ja) 1993-10-21

Similar Documents

Publication Publication Date Title
AU657781B2 (en) Novel biologically active peptide compositions and uses therefor
AU674525B2 (en) Biologically active peptides having N-terminal substitutions
AU2667192A (en) Biologically active amphiphilic peptide compositions and uses therefor
US5792831A (en) Analogues of magainin peptides containing D-amino acids
US5225399A (en) Antimicrobial amphiphilic peptides
EP0671938A4 (fr) Nouveaux peptides a activite biologique et leurs applications.
US5620954A (en) Peptide compositions and uses therefor
US5217956A (en) Composition and treatment with biologically active peptides and certain anions
WO1991012015A1 (fr) Peptides amphiphiles biologiquement actifs et procede pour inhiber la croissance de cellules cibles, de virus ou de cellules contaminees par virus
US5654274A (en) Biologically active peptides having N-terminal substitutions
EP0590044A1 (fr) Composition de peptides biologiquement actifs presentant des substitutions au niveau de leur terminaison c et traitement a l'aide de ces peptides
CA2125494A1 (fr) Traitement a l'aide d'une composition a base de peptides et d'agents de chelation biologiquement actifs
US5459237A (en) Peptide compositions and uses therefor
AU641129B2 (en) Composition and treatment with biologically active peptides and certain anions
EP0533795A1 (fr) Composition et traitement utilisant des peptides biologiquement actifs contenant des resites d'aminoacides d
AU693518B2 (en) Ion-channel forming amphiphilic peptides having n-terminal modifications
WO1991016066A1 (fr) Compositions et traitement a base de peptides biologiquement actifs et de cations toxiques
EP0559647A4 (fr) COMPOSITION ET TRAITEMENT A L'AIDE DE PEPTIDES BIOLOGIQUEMENT ACTIFS ET D'ANTIBIOTIQUES INHIBANT LA GYRASE d'ADN.

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

17P Request for examination filed

Effective date: 19921030

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): AT BE CH DE DK ES FR GB GR IT LI LU NL SE

RHK1 Main classification (correction)

Ipc: C07K 7/06

A4 Supplementary search report drawn up and despatched

Effective date: 19930906

AK Designated contracting states

Kind code of ref document: A4

Designated state(s): AT BE CH DE DK ES FR GB GR IT LI LU NL SE

17Q First examination report despatched

Effective date: 19940225

RAP1 Party data changed (applicant data changed or rights of an application transferred)

Owner name: MAGAININ PHARMACEUTICALS INC.

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE APPLICATION IS DEEMED TO BE WITHDRAWN

18D Application deemed to be withdrawn

Effective date: 19971201