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WO1994019369A1 - Treatment of cancerous growths with biologically active peptides and protease inhibitors - Google Patents

Treatment of cancerous growths with biologically active peptides and protease inhibitors Download PDF

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Publication number
WO1994019369A1
WO1994019369A1 PCT/US1994/002121 US9402121W WO9419369A1 WO 1994019369 A1 WO1994019369 A1 WO 1994019369A1 US 9402121 W US9402121 W US 9402121W WO 9419369 A1 WO9419369 A1 WO 9419369A1
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WO
WIPO (PCT)
Prior art keywords
amino acid
peptide
lys
hydrophilic
ala
Prior art date
Application number
PCT/US1994/002121
Other languages
French (fr)
Inventor
Meenhard Herlyn
Leonard S. Jacob
W. Lee Maloy
Original Assignee
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 Pharmaceuticals, Inc. filed Critical Magainin Pharmaceuticals, Inc.
Priority to AU62515/94A priority Critical patent/AU6251594A/en
Publication of WO1994019369A1 publication Critical patent/WO1994019369A1/en

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
    • A61K38/08—Peptides having 5 to 11 amino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/1703—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • A61K38/1709—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • A61K38/1751—Bactericidal/permeability-increasing protein [BPI]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
    • A61K38/10—Peptides having 12 to 20 amino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/1703—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
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/1703—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • A61K38/1709—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/55—Protease inhibitors

Definitions

  • This invention relates to the treatment of cancerous growths. More particularly, the invention relates to the treatment of cancerous growths by administering a biologically active peptide and a protease inhibitor.
  • a process for treating a cancerous growth in a host which comprises administering to a host (a) at least one biologically active amphiphilic ion channel-forming peptide or protein; and (b) at least one protease inhibitor.
  • the at least one peptide or protein and the at least one protease inhibitor are administered in amounts effective to treat a cancerous growth in a host.
  • treating a cancerous growth means that the peptide or protein and protease inhibitor inhibit, prevent, or destroy the growth of cancer cells, and/or reduces the size of or eliminates a cancerous growth.
  • An ion channel-forming peptide or protein or ionophore is a peptide or protein which increases the permeability for ions across a natural or synthetic lipid membrane.
  • B. Christensen, et al., PNAS, Vol. 85, pgs. 5072-5076 (July 1988) describes methodology which indicates whether or not a peptide has ion channel-forming properties and is therefore an ionophore.
  • an ion channel-forming peptide or ion channel-forming protein is a peptide or protein which has ion channel-forming properties as determined by the method of Christensen, et al.
  • amphiphilic peptide or protein is a peptide or protein which includes both hydrophobic and hydrophilic peptide or protein regions.
  • the ion channel-forming peptides or proteins of the present invention are generally water soluble to a concentration of at least 20 mg/ml at neutral pH in water.
  • the structure of such peptide provides for flexibility of the peptide molecule.
  • Such peptides are capable of forming an alpha-helix. 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 rodlike structure.
  • such peptides have at least 7 amino acids, and in many cases have at least 20 amino acids. In most cases, such peptides do not have in excess of 40 amino acids.
  • biologically active peptide or protein, and protease inhibitor are administered to a host
  • biologically active peptide or protein and the protease inhibitor may be administered as a single composition or in separate compositions, and the single or separate compositions may include additional materials, actives and/or inactives, in
  • Protease inhibitors which may be employed include, but are not limited to, bestatin, amastatin, and leupeptin. In one embodiment the protease inhibitor is bestatin.
  • the peptide or protein and the protease inhibitor are employed in amounts effective to inhibit and/or prevent and/or destroy a cancerous growth.
  • the protease inhibitor potentiates the action of the peptide or protein, and the peptide or protein potentiates the action of the protease inhibitor.
  • protease inhibitor means that the amount of protease inhibitor is effective to reduce the minimum effective concentration of the peptide or protein for inhibiting and/or reducing and/or destroying a cancerous growth, and the amount of peptide or protein is effective to reduce the minimum effective concentration of the
  • protease inhibitor for inhibiting and/or reducing and/or destroying a cancerous growth.
  • each of the peptide or protein and protease inhibitor is administered in an amount ineffective to treat a
  • cancerous growth in a host if administered alone to a host if administered alone to a host.
  • the peptide or protein and protease inhibitor may be administered to a host in vivo, such as, for example through systemic
  • peptides or proteins and protease inhibitors may be administered, such as intravenous or intraperitoneal administration.
  • administration such as intravenous or intraperitoneal administration.
  • peptide or protein administered intralesionally, i.e., the peptide or protein and
  • protease inhibitor are injected directly into the cancerous growth.
  • the peptide or protein may be administered in an amount of from abour 50 ⁇ g per injection to about 100 mg per injection such that the peptide or protein preferably is administered in an amount of from about 1 mg/kg to about 5 mg/kg of body weight.
  • the protease inhibotor may be administered in an amount of from about 50 ⁇ g per injection to about 100 mg per injection, preferably from about 1 mg/kg to about 5 mg/kg of body weight.
  • inhibitor may be administered in multiple doses.
  • Canceorus growths which may be treated with the peptide or protein and protease inhibitor include, but are not limited to, skin cancers, such as melanoma, squamous cell carcinoma, and basal cell carcinoma, or other cancers, such as ovarian cancers, uterine cancers, cervical cancer, and breast cancer.
  • skin cancers such as melanoma, squamous cell carcinoma, and basal cell carcinoma
  • other cancers such as ovarian cancers, uterine cancers, cervical cancer, and breast cancer.
  • the peptides or proteins, and at least one protease inhibitor are administered in combination with an acceptable pharmaceutical carrier or vehicle such as a filler, non-toxic buffer, or
  • the peptides or proteins may also be used in combination with adjuvants or compatible drugs.
  • the peptide is 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. Still more particularly, the hydrophobic amino acids are in groups of two adjacent amino acids, and each group of two hydrophobic amino acids is spaced from another group of two hydrophobic amino acids by at least one amino acid other than a hydrophobic amino acid (preferably at least two amino acids) and generally by no greater than four amino acids, and the amino acids between pairs of hydrophobic amino acids may or may not be
  • the hydrophilic amino acids are generally also in groups of two adjacent amino acids in which at least one of the two amino acids is a basic hydrophilic amino acid, with such groups of two hydrophilic amino acids being spaced from each other by at least one amino acid other than a hydrophilic amino acid (preferably at least two amino acids) and generally no greater than four amino acids, and the amino acids between pairs of hydrophilic amino acids may or may not be hydrophobic.
  • the polypeptide comprises a chain of at least four groupjs of amino acids, with each group consisting of four amino acids. 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 Ala, Cys, Phe, Gly, lle. Leu, Met, Pro, Val, Trp, Tyr, norleucine (Nle), norvaline (Nva), and cyclohexylalanine (Cha).
  • the neutral hydrophilic amino acids may be selected from the class
  • the basic hydrophilic amino acids may be selected from the class consisting of Lys, Arg, His, Orn, homoarginine (Har), 2, 4-diaminobutyric acid
  • 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. 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.
  • 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 and two hydrophilic amino acids as hereinabove noted.
  • the biologically active polypeptide may comprise a chain including at least four groups of amino acids, each containing four amino acids. Two of the four amino acids in each group are
  • hydrophobic, at least one amino acid is basic hydrophilic, and the remaining one is basic or neutral hydrophilic, 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,
  • the resulting polypeptide chain may have one of the following sequences: (X 1 ) a (A-B-C-D) n (Y 1 ) b
  • X 1 is D; C-D- or B-C-D-, Y 1 is -A or -A-B or -A-B-C
  • 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 i s -C , -C-D , -C-D-A
  • n is at least 4.
  • the peptide chain may include amino acids between the hereinabove noted groups of four amino acids provided that the spacing between such groups and the charge on the amino acids does not change the characteristics of the peptide chain which provide amphiphilicity and a positive charge and do not adversely affect the folding characteristics of the chain to that which is significantly different from one in which the hereinabove noted group of four amino acids are not spaced from each other.
  • the peptide may have amino acids extending from either end of chain.
  • the chains may have a Ser-Lys sequence before "Ala” end, and/or an Ala-Phe sequence after the "Lys" end.
  • Other amino acid sequences may also be attached to the "Ala” and/or the "Lys” end.
  • the chain in any polypeptide chain having at least four groups of amino acids of the sequence as described above, the chain may have, for example, a C-D sequence before the first A-B-C-D group.
  • other amino acid sequences may be attached to the "A” and/or the "D" end of one of these polypeptide chains.
  • there may be amino acids in the chain which space one or more groups of the hereinabove noted four amino acids from each other.
  • the peptide may be a magainin peptide.
  • a magainin peptide is either a magainin such as magainin I, II or III or an analogue or derivative thereof.
  • the magainin peptides preferably include the following basic peptide structure X 12 :
  • R 13 is a hydrophobic, neutral hydrophilic, or basic hydrophilic amino acid
  • R 14 and R 14 are hydrophobic or basic hydrophilic amino acids
  • R 15 is glutamic acid or aspartic acid, or a hydrophobic or a basic hydrophilic amino acid
  • n is 0 or 1.
  • R 13 is a hydrophobic or neutral hydrophilic amino acid
  • R 14a is a hydrophobic amino acid
  • R 15 is glutamic acid or aspartic acid.
  • a magainin peptide may include the following structure:
  • R 11 , R 12 , R 14 and R 14a are as previously defined.
  • a magainin peptide may also have the following structure:
  • R 16 where R 16 is a basic hydrophilic amino acid or
  • R 16 -R 17 where R 17 is a neutral hydrophilic amino acid, a hydrophobic amino acid, or a basic hydrophilic amino acid.
  • R 17 is a neutral hydrophilic amino acid.
  • a magainin peptide may also have the following structure:
  • X 12' Y 12 and Z 12 are as Previously defined and a is 0 or 1 and b is 0 or 1.
  • the magainin peptides may also include the following basic peptide structure X 13 :
  • the magainin peptide may aisp include the following structure
  • R 11 , R 14' R 14a' R 15' R 16' and R 17 are as hereinabove
  • n is 0 or 1, and each n may be the same or different
  • the magainin peptides general ly include at least fourteen amino acids and may include up to forty amino acids.
  • a magainin peptide preferably has 22 or 23 amino acids. Accordingly, the hereinabove described basic peptide structures of a magainin peptide may include additional amino acids at the amino end or at the carboxyl end, or at both ends.
  • magainin peptides having the following primary sequences as given in the accompanying sequence listing as well as appropriate analogues and derivatives thereof:
  • Magainin peptides are described in Proc. Natl. Acad Sci. Vol.84 pp. 5449-53 (Aug. 87).
  • magainin peptides refers to the basic magainin structure as well as derivatives and analogs thereof, including but not limited to the representative derivatives or analogs.
  • the peptide may be a PGLa peptide or an XPF peptide.
  • a PGLa peptide is either PGLa or an analogue or derivative thereof.
  • the PGLa peptides preferably include the following basic peptide structure X 14 :
  • R 11 -R 11 -R 12 - where R 11 , R 12 , R 14 , and R 17 are as previously defined.
  • the PGLa peptides generally include at least seventeen amino acids and may include as many as forty amino acids. Accordingly, the hereinabove described basic peptide structure for a PGLa peptidemay include additional amino acids at the amino end or at the carboxyl end or at both the amino and carboxyl end.
  • a PGLa peptide may have the following
  • R 11 and R 14 are as previously defined.
  • a PGLa peptide may also have the following
  • R 11 is as previously defined.
  • a PGLa peptide may also have the following structure:
  • X 14 ; Y 14 and Z 14 are as previously defined, a is 0 or 1 and b is 0 or 1.
  • An XPF peptide is either XPF or an analogue or derivative thereof.
  • the XPF peptides preferably include the following basic peptide structure X 16 :
  • R 11' R 12' R 14' R 15 and R 17 are as Previously defined and R 18 is glutamine or asparagine or a basic hydrophilic, or hydrophobic amino acid and, n is 0 or 1.
  • 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 end, or at the carboxyl end or at both the amino and carboxyl ends.
  • an XPF peptide may include the following structure:
  • 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 sequences as given in the
  • the peptide may be a CPF peptide or appropriate analogue or derivative thereof.
  • CPF peptides as well as analogues and derivatives thereof are herein sometimes referred to collectively as CPF peptides.
  • the CPF peptide may be one which includes the following basic peptide structure X 20 :
  • R 21 is a hydrophobic amino acid
  • R 22 is a hydrophobic amino acid or a basic hydrophilic amino acid
  • R 23 is a basic hydrophilic amino acid
  • R 24 is a hydrophobic or neutral hydrophilic amino acid
  • R 25 is a basic or neutral hydrophilic amino acid.
  • hydrophobic amino acids are Ala, Cys, Phe, Gly, lle, Leu, Met, Val, Trp, Tyr, norleucine (Nle), norvaline (Nva), and
  • the neutral hydrophilic amino acids are Asn, Gln, Ser, Thr, and homoserine (Hse).
  • the basic hydrophilic amino acids are Lys, Arg, His, Orn, homoarginine (Har), 2,4-diaminobutyric acid (Dbu), and
  • the CPF peptide may include only the hereinabove noted amino acids or may include additional amino acids at the amino and/or carboxyl end or both the amino and carboxyl end. In general, the peptide does not include more than 40 amino acids.
  • the CPF peptides including the above basic structure preferably have from 1 to 4 additional amino acids at the amino end.
  • R 21 , R 22 and R 25 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 additional amino acids.
  • 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 20 is
  • Preferred peptides may be represented by the following structural formula
  • CPF peptides which may be employed, some of which have been described in the literature, include the following sequenrps as given in the accompanying sequence listing: (SEQ ID NO: 14)
  • the peptide may include one of the following basic structures X 3 1 through X 37 wherein:
  • X 31 is -[R 31 -R 32 -R 32 -R 33 -R 31 -R 32 -R 32 ]- n;
  • X 32 is -[R 32 -R 32 -R 33 -R 31 -R 32 -R 32 -R 31 ]- n;
  • X 33 is -[R 32 -R 33 -R 31 -R 32 -R 32 -R 31 -R 32 ]- n;
  • X 34 is -[R 33 -R 31 -R 32 -R 32 -R 31 -R 32 -R 32 ]- n;
  • X 35 is -[R 31 -R 32 -R 32 -R 31 -R 32 -R 32 -R 33 ]- n;
  • X 36 is -[R 32 -R 32 -R 31 -R 32 -R 32 -R 33 -R 31 ]- n;
  • X 37 is -[R 32 -R 31 -R 32 -R 32 -R 33 -R 31 -R 32 ]- n;
  • R 31 is a basic hydrophilic amino acid
  • R 32 is a hydrophobic amino acid
  • R 33 is a neutral hydrophilic, basic hydrophilic, or hydrophobic amino acid
  • n is from 2 to 5.
  • the basic hydrophilic amino acids may be selected from the class consisting of Lys, Arg, His, Orn, homoarginine (Har),
  • the hydrophobic amino acids may be selected from the class consisting of Ala, Cys, Phe, Gly, lle, Leu, Met, Pro, Val, Trp and Tyr, norleucine (Nle), norvaline (Nva), and cyclohexylalanine (Cha) .
  • the neutral hydrophilic amino acids may be selected from the class consisting of Asn, Gln, Ser, Thr, and homoserine (Hse).
  • the peptide when the peptide includes the structure X 31 , the peptide may include the following structure:
  • Y 31 -X 31 wherein X 31 is as hereinabove described, and Y 31 is:
  • the peptide when the peptide includes the structure X 31 , the peptide may include the following structure:
  • the peptide may include the following structure:
  • the peptide may include the following structure:
  • the peptide when the peptide includes the structureX 32 , the peptide may include the following structure:
  • the peptide may include the following structure:
  • the peptide when the peptide includes the structure X 33 , the peptide may include the following structure:
  • the peptide when the peptide includes the structure X 33 , the peptide may include the following structure:
  • the peptide may include the following structure:
  • the peptide when the peptide includes the structure X 34 , the peptide may include the following structure:
  • the peptide when the peptide includes the structure X 34 , the peptide may include the following structure:
  • the peptide may include the following structure:
  • the peptide when the peptide includes the structure X 35 , the peptide may include the following structure:
  • Y 35 -X 35 wherein X 35 is as hereinabove described, and Y 35 is:
  • the peptide when the peptide includes the structure X 35 , the peptide may include the following structure:
  • the peptide may include the following structure:
  • the peptide when the peptide includes the structure X 36 .
  • the peptide may include the following structure:
  • the peptide when the peptide included the structure X 36, the peptide may include the following structure:
  • the peptide may include the following structure:
  • the peptide when the peptide includes the structure X 37 , the peptide may includes the structure Y 37 -X 37 , wherein X 37 is as hereinabove described, and Y 37 is:
  • the peptide when the peptide includes the structure X 37 , the peptide may include the following structure:
  • X 37 - Z 37 wherein X 37 is as hereinabove described, and Z 37 is: (i) R 32 ;
  • the peptide may include the following structure:
  • n is 3, and most preferably the peptide is of one of the following structures as given in the
  • Lys lle Ala (Lys lle Ala Gly Lys lle Ala) 3 (SEQ ID NO: 69)
  • Xaa is p-aminophenylalanme.
  • amphiphilic peptide includes the following basic structure X 40 :
  • R 31 , R 32 , and R 33 are as hereinabove described, and R 34 is a basic hydrophilic or hydrophobic amino acid.
  • the peptide may include the following structure:
  • Y 40 -X 40' wherein X 40 is as hereinabove described, and Y 40 is:
  • the peptide may include the following structure:
  • X 40 -Z 40 wherein X 40 is as hereinabove-described and Z 40 is:
  • Y 40 ) a -X 40 -(Z 40 ) b wherein Y 40 and Z 40 are as previously defined, a is 0 or 1, and b is 0 or 1.
  • the peptide has the following structural formula as given in the accompanying sequence listing:
  • the peptide has the following structural formula as given in the accompanying sequence l i sting :
  • the peptide has one of the one of the following structural formulae as given in the
  • the peptide may include the following structural formula:
  • n is from 2 to 5.
  • n is 3, and the peptide has the following structural formula:
  • the peptide may include the following structural formula: -(Lys Phe Ala Lys Lys Phe Ala)- n
  • n is from 2 to 5.
  • n 3
  • the peptide has the following structural formula:
  • the peptide may include the following structural formula:
  • n is from 2 to 5.
  • n is 3, and the peptide has the following structural formula:
  • the peptide may be selected from the group consisting of the following structural formulae as given in the accompanying sequence listing:
  • the peptide may be a cecropin or sarcotoxin.
  • cecropins includes the basic structure as well as analogues and derivatives thereof.
  • the cecropins and analogues and derivatives thereof are described in Ann. Rev. Microbiol. 1987, Vol. 41, pages 103-26, in particular page 108, and in Christensen, et al., PNAS Vol. 85, pgs. 5072-76, which are hereby incorporated by
  • amphiphilic peptide includes the following basic structure X 50 :
  • R 41 is a hydrophobic amino acid
  • R 42 is a basic hydrophilic or neutral hydrophilic amino acid.
  • the peptide includes the basic structure
  • Y 50 -X 50 wherein X 50 is as hereinabove described and Y 50 is:
  • R 41 is leucine.
  • R 42 is lysine.
  • Representative examples of peptides in accordance with this aspect of the present invention include those having the following structures:
  • amphiphilic peptide includes the following basic structure X 50 :
  • R 4 1 is leucine. In another embodiment, R 42 is lysine.
  • the peptide includes the basic structure
  • X 52 is as hereinabove described, and Y 52 is:
  • the peptide may have the following structure; Lys Lys Leu Leu Lys Lys Leu Lys Lys Leu Lys Lys Leu
  • the peptide includes the basic structure
  • the peptide may have the following structure:
  • the peptide may include the structure:
  • a is 0 or 1
  • b is 0 or 1.
  • the peptide includes the following basic structure X 54 :
  • R 41 and R 42 are as hereinabove described, and R 43 is a netural hydrophilic amino acid.
  • the peptide may have the following structure:
  • the peptide may have the following
  • the peptide includes the following basic structure X 56 : R 41 -R 42 -R 41 -R 41 -R 42 -R 42 -R 41 -R 41 -R 42 -R 44 , wherein R 41 and R 42 are as hereinabove described, and R 44 is a neutral hydrophilic amino acid or proline.
  • the peptide may include the following
  • Y 56 is:
  • R 41 and R 42 are as hereinabove described.
  • the peptide may include the structure:
  • the peptide may have one of the following structures:
  • the peptide may have the structure
  • the peptide includes the following basic structure X 58 :
  • the peptide may include the structure Y 58 -X 58, wherein X 58 is as hereinabove described, and
  • Y 58 is:
  • the peptide includes the structure X 58 -Z 58 , wherein X 58 is as hereinabove described, and Z 58 is:
  • the peptide has the following structure:
  • the peptide may have the structure
  • the peptide includes the following basic structure X 60 ;
  • the peptide may have the following structure:
  • the peptide may include the structure X 60 -Z 60 , wherein X 60 is as hereinabove described, and Z 60 is:
  • the peptide has a structure selected from the group consisting of:
  • the peptide has the structure (a), and a representative example of such a structure is (SEQ ID NO: 107), which is given in the accompanying sequence listing.
  • the peptide has the structure (b), and representative example of such a structure is (SEQ ID NO: 108), which is given in the accompanying sequence listing.
  • the peptide has the structure (c), and a representative example of such a structure is (SEQ ID NO: 109) as given the accompanying sequence listing.
  • the peptide has the structure (d), and a representative example of such a structure is (SEQ ID NO: 110) as given in the accompanying sequence listing.
  • the peptide has the structure (e), and representative examples of such a structure are (SEQ ID NO: 111) and (SEQ ID NO: 112) as given in the accompanying sequence listing.
  • the peptide has the following structural formula:
  • the peptide is melittin.
  • Melittin is an amphipathic peptide consisting of 26 amino acid residues, and is isolated from honeybee (Apis mellifera) venom.
  • the peptide is known to be cytolytic. See Habermann, et al.,
  • Melittin has the following structural formula as represented by the three-letter amino acid code:
  • the peptide purified in accordance with the present invention is an apidaecin.
  • apidaecin as used herein includes the basic structure as well as analogues and
  • the peptide may be an amide - or carboxy-terminated peptide represented by the following structural formula, and the numbers below each amino acid residue refer to the position of the residue in the peptide:
  • the peptide may be an analogue of such peptide wherein at least one of amino acid residues 1 through 7, 9, 11, 12, 14, 16, or 18 is deleted from the peptide.
  • amino acid residues 1, 3, 7, 9, 11, 12, 14, 16, or 18 is deleted from the peptide.
  • amino acid residues 1 through 3, 1 through 4, 1 through 5, 1 through 6, and 1 through 7 are deleted from the peptide.
  • amino acid residues 1 through 3 or 1 through 4 are deleted from the peptide, and such peptides have the following structural formulae:
  • the peptide includes the following structural formula X 62 :
  • R 41 is a hydrophobic amino acid
  • R 42 is a basic
  • R 41 is leucine, and in another embodiment, R 42 is lysine.
  • the peptide has the following structure:
  • the peptide includes the following structural formula X 64 :
  • R 41 is a hydrophobic amino acid
  • R 42 is a basic
  • R 41 is leucine, and in another embodiment, R 42 is lysine.
  • the peptide has the following structural formula:
  • the peptide includes the following
  • R 41 is a hydrophobic amino acid
  • R 42 is a basic
  • hydrophilic or neutral hydrophilic amino acid hydrophilic or neutral hydrophilic amino acid
  • the peptide may include the following
  • X 66 -Z 66 wherein X 66 is as hereinabove described and Z 66 is:
  • R 41 is leucine, and in another embodiment, R 42 is lysine.
  • the peptide has the following structural formula:
  • amphiphilic peptide or protein may be an ion channel-forming peptide or protein.
  • Ion channel-forming proteins or peptides which may be employed include defensins, also known as human neutrophil antimicrobial peptides (HNP), major basic protein (MBP) of eosinophils, bactericidal permeability-increasing protein (BPT). and a pore-forming cytotoxin called variously perforin, cytolysin, or pore-forming protein.
  • defensins also known as human neutrophil antimicrobial peptides (HNP), major basic protein (MBP) of eosinophils, bactericidal permeability-increasing protein (BPT).
  • BPT bactericidal permeability-increasing protein
  • 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 (1985).
  • 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. 14891-14894 (1987).
  • Perforin is described in Henkart, et al., J. Exp. Med., 160 75 (1984), and in Podack, et al., J. Exp. Med., 160:695 (1984). The above articles are hereby incorporated by reference.
  • ion channel-forming proteins includes the basic
  • the peptides may be produced by known techniques and obtained in substantially pure form.
  • the peptides may be synthesized on an automatic peptide 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 codons encoding the amino acids are known to those skilled in the art, and thus one may construct DNA encoding any of the peptides by accepted techniques, and clone such DNA into an expression vehicle such as, for example, a plasmid, and transfect such an expression vehicle into a cell which will express the peptide.
  • the peptide or protein and protease inhibitor may be any suitable peptide or protein and protease inhibitor.
  • chemotherapeutic agents such as, but not limited to, cyclophosphamide, cisplatin, doxorubicin,
  • the peptide or protein and protease inhibitor may be administered before, during, or after radiation treatment, chemotherapy, or
  • the administration of the peptides or proteins and protease inhibitor during surgery or radiation treatment may be advantageous in inhibiting, preventing, and/or destroying potential "loose" malignant cells capable of colonizing other sites.
  • Example The following melanoma cel l lines are seeded in 200 ul of 2 . 0% tumor medium at a concentration of 1.5 x 10 4 cells per well in a
  • each amino acid residue is a D-amino acid residue
  • triplicate a concentration of 1.0 ug/ml.
  • bestatin is added in triplicate at a concentration in 2% medium of 70 ug/ml.
  • D-(SEQ ID NO: 117) is added at a concentration of 1.0 ug/ml and bestatin is added at a concentration 70 ug/ml in 2% medium in triplicate.
  • a control group has neither peptide nor bestatin added to the cells.
  • Detached cells are harvested on a Tomtec cell harvester, and
  • radioactive counts are measured from the filters on a Packard counter.
  • the radioactive counts for each cell line contacted with the peptides is given in Table I below.
  • % decrease is based upon the difference between the results with bestatin only and the results with D-(SEQ ID NO: 117) and bestatin.
  • the peptide or protein and protease inhibitor may be employed for treating a wide variety of hosts.
  • the host may be a human or non-human animal.
  • the peptide or protein and the protease inhibitor may be employed together in a single
  • composition or in separate compositions.
  • protease inhibitor and the peptide or protein may be delivered or administered in different forms.
  • the peptide or protein and protease inhibitor 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.
  • 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 or protein and protease inhibitor may also be used in combination with adjuvants, or compatible drugs where such a combination is seen to be desirable or advantageous in treating a cancerous growth.
  • the peptide(s) or protein of the present invention may be administered to a host; in particular an animal, in an effective amount for treating a cancerous growth in conjunction with a protease inhibitor for potentiating the activity of the peptide or protein.
  • ADDRESSEE Carella, Byrne, Bain, Gilfillan,
  • NAME/KEY Magainin II peptide.
  • NAME/KEY magainin peptide
  • NAME/KEY magainin peptide
  • NAME/KEY magainin peptide

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Abstract

Compositions including biologically active peptides and protease inhibitors are useful as pharmaceuticals in the treatment of cancer.

Description

TREATMENT OF CANCEROUS GROWTHS WITH
BIOLOGICALLY ACTIVE PEPTIDES AND PROTEASE INHIBITORS
This invention relates to the treatment of cancerous growths. More particularly, the invention relates to the treatment of cancerous growths by administering a biologically active peptide and a protease inhibitor.
In accordance with an aspect of the present invention, there is provided a process for treating a cancerous growth in a host which comprises administering to a host (a) at least one biologically active amphiphilic ion channel-forming peptide or protein; and (b) at least one protease inhibitor. The at least one peptide or protein and the at least one protease inhibitor are administered in amounts effective to treat a cancerous growth in a host.
The term "treating a cancerous growth" as used herein means that the peptide or protein and protease inhibitor inhibit, prevent, or destroy the growth of cancer cells, and/or reduces the size of or eliminates a cancerous growth.
An ion channel-forming peptide or protein or ionophore is a peptide or protein which increases the permeability for ions across a natural or synthetic lipid membrane. B. Christensen, et al., PNAS, Vol. 85, pgs. 5072-5076 (July 1988) describes methodology which indicates whether or not a peptide has ion channel-forming properties and is therefore an ionophore. As used herein an ion channel-forming peptide or ion channel-forming protein is a peptide or protein which has ion channel-forming properties as determined by the method of Christensen, et al.
An amphiphilic peptide or protein is a peptide or protein which includes both hydrophobic and hydrophilic peptide or protein regions.
The ion channel-forming peptides or proteins of the present invention are generally water soluble to a concentration of at least 20 mg/ml at neutral pH in water. In addition, the structure of such peptide provides for flexibility of the peptide molecule. Such peptides are capable of forming an alpha-helix. 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 rodlike structure.
In general, such peptides have at least 7 amino acids, and in many cases have at least 20 amino acids. In most cases, such peptides do not have in excess of 40 amino acids.
In accordance with an aspect of the present invention wherein the biologically active peptide or protein, and protease inhibitor are administered to a host, such biologically active peptide or protein and the protease inhibitor may be administered as a single composition or in separate compositions, and the single or separate compositions may include additional materials, actives and/or inactives, in
addition to the peptide and/or protein and protease inhibitor.
Protease inhibitors which may be employed include, but are not limited to, bestatin, amastatin, and leupeptin. In one embodiment the protease inhibitor is bestatin.
In employing both an ion channel-forming biologically active amphiphilic peptide or an ion channel-forming protein, and a protease inhibitor, whether administered or prepared in a single composition, or in separate compositions, the peptide or protein and the protease inhibitor are employed in amounts effective to inhibit and/or prevent and/or destroy a cancerous growth. In effect, the protease inhibitor potentiates the action of the peptide or protein, and the peptide or protein potentiates the action of the protease inhibitor. The term "potentiate," as employed herein, means that the amount of protease inhibitor is effective to reduce the minimum effective concentration of the peptide or protein for inhibiting and/or reducing and/or destroying a cancerous growth, and the amount of peptide or protein is effective to reduce the minimum effective concentration of the
protease inhibitor for inhibiting and/or reducing and/or destroying a cancerous growth. In one embodiment, each of the peptide or protein and protease inhibitor is administered in an amount ineffective to treat a
cancerous growth in a host if administered alone to a host.
The peptide or protein and protease inhibitor may be administered to a host in vivo, such as, for example through systemic
administration, such as intravenous or intraperitoneal administration. Also, the peptides or proteins and protease inhibitors may be
administered intralesionally, i.e., the peptide or protein and
protease inhibitor are injected directly into the cancerous growth. The peptide or protein may be administered in an amount of from abour 50 μg per injection to about 100 mg per injection such that the peptide or protein preferably is administered in an amount of from about 1 mg/kg to about 5 mg/kg of body weight. The protease inhibotor may be administered in an amount of from about 50 μg per injection to about 100 mg per injection, preferably from about 1 mg/kg to about 5 mg/kg of body weight. The peptide or protein and the protease
inhibitor may be administered in multiple doses.
Canceorus growths which may be treated with the peptide or protein and protease inhibitor include, but are not limited to, skin cancers, such as melanoma, squamous cell carcinoma, and basal cell carcinoma, or other cancers, such as ovarian cancers, uterine cancers, cervical cancer, and breast cancer.
The peptides or proteins, and at least one protease inhibitor, are administered in combination with an acceptable pharmaceutical carrier or vehicle such as a filler, non-toxic buffer, or
physiological saline solution. The peptides or proteins may also be used in combination with adjuvants or compatible drugs.
In one embodiment, the peptide is 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. Still more particularly, the hydrophobic amino acids are in groups of two adjacent amino acids, and each group of two hydrophobic amino acids is spaced from another group of two hydrophobic amino acids by at least one amino acid other than a hydrophobic amino acid (preferably at least two amino acids) and generally by no greater than four amino acids, and the 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 amino acids in which at least one of the two amino acids is a basic hydrophilic amino acid, with such groups of two hydrophilic amino acids being spaced from each other by at least one amino acid other than a hydrophilic amino acid (preferably at least two amino acids) and generally no greater than four amino acids, and the amino acids between pairs of hydrophilic amino acids may or may not be hydrophobic.
In accordance with a particularly preferred embodiment, the polypeptide comprises a chain of at least four groupjs of amino acids, with each group consisting of four amino acids. 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 Ala, Cys, Phe, Gly, lle. Leu, Met, Pro, Val, Trp, Tyr, norleucine (Nle), norvaline (Nva), and cyclohexylalanine (Cha). The neutral hydrophilic amino acids may be selected from the class
consisting of Asn, Gln, Ser, Thr and homoserine (Hse). The basic hydrophilic amino acids may be selected from the class consisting of Lys, Arg, His, Orn, homoarginine (Har), 2, 4-diaminobutyric acid
(Dbu), and p-aminophenylalanine.
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. In one embodiment, 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. 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.
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 and two hydrophilic amino acids as hereinabove noted.
Thus the biologically active polypeptide may comprise a chain including at least four groups of amino acids, each containing four amino acids. Two of the four amino acids in each group are
hydrophobic, at least one amino acid is basic hydrophilic, and the remaining one is basic or neutral hydrophilic, with the polypeptide chain preferably having at least 20 amino acids but no greater than 50 amino acids.
In one embodiment, 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 hydrophobic amino acids, one of C or D is a basic hydrophilic amino acid, and the other of C or D is basic or neutral hydrophilic amino acid. The resulting polypeptide chain, therefore, may have one of the following sequences: (X1)a(A-B-C-D)n(Y1)b
(X2)a(B-C-D-A)n(Y2)b
(X3)a(C-D-A-B)n(Y3)b
(X4)a(D-A-B-C)n(Y4)b
wherein X1 is D; C-D- or B-C-D-, Y1 is -A or -A-B or -A-B-C
X2 is A-, D-A- or C-D-A-
Y2 is -B, -B-C or B-C-D
X3is B-, A-B-, D-A-B- Y3 i s -C , -C-D , -C-D-A
X4 i s C- , B-C- , A-B-C-
Y4 i s -D , -D-A , -D-A-B
a is 0 or 1; b is 0 or 1
and n is at least 4.
It is to be understood that the peptide chain may include amino acids between the hereinabove noted groups of four amino acids provided that the spacing between such groups and the charge on the amino acids does not change the characteristics of the peptide chain which provide amphiphilicity and a positive charge and do not adversely affect the folding characteristics of the chain to that which is significantly different from one in which the hereinabove noted group of four amino acids are not spaced from each other.
As representative examples of such peptides, there may be mentioned.
I Ala-Phe-Ser-Lys-Ala-Phe-Ser-Lys-Ala-Phe-Ser- Lys-Ala-Phe-Ser-Lys-Ala-Phe-Ser-Lys
(SEQ ID NO:1)
II Ala-Phe-Ser-Lys-Ala-Phe-Ser-Lys-Ala-Phe-Ser- Lys-Ala-Phe-Ser-Lys-Ala-Phe-Ser-Lys-Ala-Phe- Ser-Lys. (SEQ ID NO: 2)
III Phe-Ser-Lys-Ala-Phe-Ser- Lys-Ala-Phe-Ser-Lys-Ala- Phe-Ser-Lys-Ala- (SEQ ID NO: 3)
IV Ser-Lys-Ala-Phe-Ser-Lys-Ala- Phe-Ser-Lys-Λla-Phe-Ser-Lys-Ala- Phe-Ser-Lys-Ala-Phe- (SEQ ID NO: 4)
V Lys-Ala-Phe-Ser-Lys-Ala-Phe-Ser-Lys-Ala-Phe-Ser- Lys-Ala-Phe-Ser (SEQ ID NO: 5)
The peptide may have amino acids extending from either end of chain. For example, the chains may have a Ser-Lys sequence before "Ala" end, and/or an Ala-Phe sequence after the "Lys" end. Other amino acid sequences may also be attached to the "Ala" and/or the "Lys" end. Similarly, in any polypeptide chain having at least four groups of amino acids of the sequence as described above, the chain may have, for example, a C-D sequence before the first A-B-C-D group. Also other amino acid sequences may be attached to the "A" and/or the "D" end of one of these polypeptide chains. Also there may be amino acids in the chain which space one or more groups of the hereinabove noted four amino acids from each other.
In accordance with another embodiment, the peptide may be a magainin peptide.
A magainin peptide is either a magainin such as magainin I, II or III or an analogue or derivative thereof. The magainin peptides preferably include the following basic peptide structure X12:
-- R11- R11- R12-R13-R11R14-R12-R11-
R14-R12-R11-R11-R11-R14a-(R15)n-R14a-R14 -- wherein R11 is a hydrophobic amino acid, R12 is a basic
hydrophilic amino acid; R13 is a hydrophobic, neutral hydrophilic, or basic hydrophilic amino acid; R14 and R14 are hydrophobic or basic hydrophilic amino acids; R15 is glutamic acid or aspartic acid, or a hydrophobic or a basic hydrophilic amino acid, and n is 0 or 1. In a preferred embodiment, R13 is a hydrophobic or neutral hydrophilic amino acid, R14a is a hydrophobic amino acid, and R15 is glutamic acid or aspartic acid.
Thus, for example, a magainin peptide may include the following structure:
-Y12-X12- where X12 is the hereinabove described basic peptide structure and Y12 is
(i) R12
(ii) R14a-R12
(iii) R11-R14a-R12
(iv) R14-R11-R14a-R12
where R11, R12, R14 and R14a are as previously defined.
A magainin peptide may also have the following structure:
-X12-Z12- wherein X12 is as previously defined and Z12 is:
(i) R16 where R16 is a basic hydrophilic amino acid or
asparagine or glutamine.
(ii) R16-R17 where R17 is a neutral hydrophilic amino acid, a hydrophobic amino acid, or a basic hydrophilic amino acid.
Preferably, R17 is a neutral hydrophilic amino acid.
A magainin peptide may also have the following structure:
(Y12)a-X12-( Z12) b
where X12' Y12 and Z12 are as Previously defined and a is 0 or 1 and b is 0 or 1.
The magainin peptides may also include the following basic peptide structure X13:
--R14-R11-R14a-R12-R11-R11-R12-R13-
R11-R14-R12-R11-R11-R12-, wherein R11, R12, R13, R14, and R14a are amino acids as hereinabove described.
The magainin peptide may aisp include the following structure
X13-Z13; wherein X13 is the hereinabove described basic peptide structure and Z13 is
(R11)n-(R11)n-(R11)n-(R14a)n-(R15)n-(R14a)n-(R14)n-(R16)n-(R17)n wherein R11 , R14' R14a' R15' R16' and R17 are as hereinabove
described, and n is 0 or 1, and each n may be the same or different
The magainin peptides general ly include at least fourteen amino acids and may include up to forty amino acids. A magainin peptide preferably has 22 or 23 amino acids. Accordingly, the hereinabove described basic peptide structures of a magainin peptide may include additional amino acids at the amino end or at the carboxyl end, or at both ends.
As representative examples of such magainin peptides, there may be mentioned peptides having the following primary sequences as given in the accompanying sequence listing as well as appropriate analogues and derivatives thereof:
(a) (SEQ ID NO:6) (OH) or (NH2)
(Magainin I) (b) (SEQ ID NO:7) (OH) or (NH2)
(Magainin II)
(c) (SEQ ID NO: 8) (OH) or (NH2)
(Magainin III)
The following are examples of peptide derivatives or analogs of the basic structure:
(d) (SEQ ID NO:9) (OH) or (NH2)
(e) (SEQ ID NO:10) (OH) or (NH2)
(f) (SEQ ID NO:11) (OH) or (NH2)
Magainin peptides are described in Proc. Natl. Acad Sci. Vol.84 pp. 5449-53 (Aug. 87). The term "magainin peptides" as used herein refers to the basic magainin structure as well as derivatives and analogs thereof, including but not limited to the representative derivatives or analogs.
In accordance with a further embodiment, the peptide may be a PGLa peptide or an XPF peptide.
A PGLa peptide is either PGLa or an analogue or derivative thereof. The PGLa peptides preferably include the following basic peptide structure X14:
- R11-R17-R12-R11-R14-R14-R11-
R11-R14-R12-R11-R11-R12-R11-
R11-R11-R12- where R11, R12, R14, and R17 are as previously defined.
The PGLa peptides generally include at least seventeen amino acids and may include as many as forty amino acids. Accordingly, the hereinabove described basic peptide structure for a PGLa peptidemay include additional amino acids at the amino end or at the carboxyl end or at both the amino and carboxyl end.
Thus, for example, a PGLa peptide may have the following
structure:
-Y14-X14- where X14 is as previously defined and
Y14 is
(i) R 11; (ii) R14-R11
where R11 and R14 are as previously defined.
For example, a PGLa peptide may also have the following
structure:
-X14-Z14- where X1 4 i s as previously defined; and Z1 4 i s :
( i ) R11 ; or
( ii ) R11 -R11
where R11 is as previously defined.
A PGLa peptide may also have the following structure:
(Y14)a -X14-( Z14) b
where X14; Y14 and Z14 are as previously defined, a is 0 or 1 and b is 0 or 1.
An XPF peptide is either XPF or an analogue or derivative thereof. The XPF peptides preferably include the following basic peptide structure X16:
-- R11-R17-R12-R11-R14-R18-R17-
R11-R14-R12-R11-R11-R12- R11-R11- R11-R12-(R15)n-R11--, wherein
R11' R12' R14' R15 and R17 are as Previously defined and R18 is glutamine or asparagine or a basic hydrophilic, or hydrophobic amino acid and, n is 0 or 1.
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 end, or at the carboxyl end or at both the amino and carboxyl ends.
Thus, for example, an XPF peptide may include the following structure:
-Y16-X16- where X16 is as previously defined and Y16 is
(i) R11 or
(ii) R14-R11 where R11 and R14 are as previously defined.
An XPF peptide may include the following structure:
-X16-Z16- where X16 is as previously defined and Z16 is
(i) R11; or
(ii) R11-R18; or
(iii) R11-R18-Proline; or
(iv) R11-R18-Proline-R12
An XPF peptide may also have the following structure:
(Y16)a-X16-(Z16)b where X16, Y16 and Z16 are as previously defined: a is 0 or 1 and b is 0 or 1.
Preferred are XPF or PGLa peptides, which are characterized by the following primary amino acid sequences as given in the
accompanying sequence listing:
PGLa : (SEQ ID NO: 12) (NH2)
XPF : (SEQ ID NO: 13)
A review of XPF and PGLa can be found in Hoffman et al, EMBO J. 2:711-714, 1983; Andreu, et al, J. Biochem. 149:531-535, 1985; Gibson, et al J. Biol. Chem. 261:5341-5349, 1986; and Giovannini, et al,
Biochem J. 243:113-120, 1987.
In accordance with yet another embodiment, the peptide may be a CPF peptide or appropriate analogue or derivative thereof.
CPF peptides as well as analogues and derivatives thereof are herein sometimes referred to collectively as CPF peptides.
The CPF peptide may be one which includes the following basic peptide structure X20:
-R21-R21-R22-R22-R21-R21-R23-R21-
-R21-R21-R23-R21-R21-R24-R25-R21- wherein R21 is a hydrophobic amino acid;
R22 is a hydrophobic amino acid or a basic hydrophilic amino acid;
R23 is a basic hydrophilic amino acid; R24 is a hydrophobic or neutral hydrophilic amino acid; and
R25 is a basic or neutral hydrophilic amino acid.
The hereinabove basic structure is hereinafter symbolically indicated as X20.
The hydrophobic amino acids are Ala, Cys, Phe, Gly, lle, Leu, Met, Val, Trp, Tyr, norleucine (Nle), norvaline (Nva), and
cyclohexylalanine (Cha).
The neutral hydrophilic amino acids are Asn, Gln, Ser, Thr, and homoserine (Hse).
The basic hydrophilic amino acids are Lys, Arg, His, Orn, homoarginine (Har), 2,4-diaminobutyric acid (Dbu), and
p-aminophenylalanine.
The CPF peptide may include only the hereinabove noted amino acids or may include additional amino acids at the amino and/or carboxyl end or both the amino and carboxyl end. In general, the peptide does not include more than 40 amino acids.
The CPF peptides including the above basic structure preferably have from 1 to 4 additional amino acids at the amino end.
Accordingly, such preferred peptides may be represented by the structural formula:
Y20 - X20 - wherein X20 is the hereinabove described basic peptide structure and Y20 is
(i) R25-, or
(ii) R22-R25-; or
(iii) R21-R22-R25; or
(iv) R22-R21-R22-R25; Preferably
Glycine - R21-R22-R25.
wherein R21, R22 and R25 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 additional amino acids. In a preferred embodiment, the basic structure may have from 1 to 7 additional amino acids at the carboxyl end, which may be represented as follows:
-X20 - Z20 wherein
X is the hereinabove defined basic peptide structure and Z20 is
(i) R21-, or
(ii) R21-R21-; or
(iii) R21-R21-R24; or
(iv) R21-R21-R24-R24; or
(v) R21-R21-R24-R24-R26; or
(vi) R21-R21-R24-R24-R26-Gln; or
(vii) R2- R21-R24 -R-24 -R26-Gln-Gln, wherein R21 and R24 are previously defined, and R26 is proline or a hydrophobic amino acid.
Preferred peptides may be represented by the following structural formula
(Y20)a - X20 - (Z20)b wherein X20, Y20 and Z20 are as previously defined and a is 0 or 1 and b is 0 or 1.
Representative examples of CPF peptides which may be employed, some of which have been described in the literature, include the following sequenrps as given in the accompanying sequence listing: (SEQ ID NO: 14)
(SEQ ID NO: 15)
(SEQ ID NO: 16)
(SEQ ID NO: 17)
(SEQ ID NO: 18)
(SEQ ID MO: 19)
(SEQ ID NO: 20)
(SEQ ID NO: 21)
(SEQ ID NO: 22)
(SEQ ID NO: 23)
(SEQ ID NO: 24)
(SEQ ID NO: 25) ( SEQ ID NO : 26 )
A review of the CPF peptides can be found in Richter, K., Egger, R., and Kreil (1986) J. Biol. Chem 261, 3676-3680; Wakabayashi, T., Kato, H., and Tachibaba, S. (1985) Nucleic Acids Research 13, 1817-1828; Gibson, B.W., Poulter, L., Williams, D.H., and Maggio, J.E. (1986) J. Biol. Chem 261, 5341-5349.
In accordance with yet another embodiment, the peptide may include one of the following basic structures X3 1 through X37 wherein:
X31 is -[R31-R32-R32-R33-R31-R32-R32]-n;
X32 is -[R32-R32-R33-R31-R32-R32-R31]-n;
X33 is -[R32-R33-R31-R32-R32-R31-R32]-n;
X34 is -[R33-R31-R32-R32-R31-R32-R32]-n;
X35 is -[R31-R32-R32-R31-R32-R32-R33]-n;
X36 is -[R32-R32-R31-R32-R32-R33-R31]-n; and
X37 is -[R32-R31-R32-R32-R33-R31-R32]-n;
wherein R31 is a basic hydrophilic amino acid, R32 is a hydrophobic amino acid, R33 is a neutral hydrophilic, basic hydrophilic, or hydrophobic amino acid, and n is from 2 to 5.
The basic hydrophilic amino acids may be selected from the class consisting of Lys, Arg, His, Orn, homoarginine (Har),
2,4-diaminobutyric acid (Dbu), and p-aminophenylalanine.
The hydrophobic amino acids may be selected from the class consisting of Ala, Cys, Phe, Gly, lle, Leu, Met, Pro, Val, Trp and Tyr, norleucine (Nle), norvaline (Nva), and cyclohexylalanine (Cha) .
The neutral hydrophilic amino acids may be selected from the class consisting of Asn, Gln, Ser, Thr, and homoserine (Hse).
In accordance with one embodiment, when the peptide includes the structure X31 , the peptide may include the following structure:
Y31-X31, wherein X31 is as hereinabove described, and Y31 is:
(i) R32;
(ii) R32- R32;
(iii) R31-R32-R32;
(iv) R33-R31-R32-R32;
(V) R32-R33-R31-R32-R 32; or (vi) R32-R32-R33-R31-R32-R32, wherein R31, R32, and R33 are as hereinabove described
In accordance with another embodiment, when the peptide includes the structure X31 , the peptide may include the following structure:
X31-Z31, wherein X31 is as hereinabove described, and Z31 is:
(i) R31;
(ii) R31-R32;
(iii) R31-R32-R32;
(iv) R31-R32-R32-R33;
(v) R31-R32-R32-R33-R31; or
(vi) R31-R32-R32-R33-R31-R32.
In accordance with yet another embodiment, the peptide may include the following structure:
(Y31)a-X31-(Z31)b' wherein Y31 and Z31 are as Previously defnied, a is 0 or 1, and b is 0 or 1.
When the peptide includes the structure X32, the peptide may include the following structure:
Y32 - X32' wherein X32 is as hereinabove described, and Y32 is:
( i ) R31 ;
( ii ) R32-R31 ;
( i ii ) R32-R32-R31 ;
( iv) R3 1-R32-R32-R3 1 ;
(v ) R33-R31-R32-R32-R31 ; or
( vi ) R32-R33-R31-R32-R32-R31.
In another embodiment, when the peptide includes the structureX32, the peptide may include the following structure:
X32 - Z32, wherein X32 is as hereinabove described, and Z32 is:
(i) R32;
( i i ) R32-R32;
(iii) R32-R32-R33;
(iv) R32-R32-R33-R31;
(v) R32-R32-R33-R31-R32; or
(vi) R32-R32-R33-R31-R32-R32. in accordance with yet another embodiment, the peptide may include the following structure:
(Y32)a - X32 - (Z32)b, wherein Y32 and Z32 are as Previously defined, a is 0 or 1, and b is 0 or 1.
In accordance with another embodiment, when the peptide includes the structure X33, the peptide may include the following structure:
Y33 - X33 wherein X33 is as hereinabove described, and Y33 is:
(i) R32;
(ii) R31-R32;
(iii) R32-R31-R32;
( iv ) R32-R32-R31-R32 ;
( v ) R31- R32-R32-R31-R32 ; or
(vi) R33-R31-R32-R32-R31-R32, wherein R31, R32, and R33 are as hereinabove described.
In accordance with another embodiment, when the peptide includes the structure X33, the peptide may include the following structure:
X33 - Z33 wherein X33 is as hereinabove described, and Z33 is:
(i) R32;
(ii) R32-R33;
(iii) R32-R33-R31;
( iv ) R32-R32-R31-R32 ;
( v ) R31-R32-R32-R31-R32 ; or
(vi) R33-R31-R32-R32-R31-R32,
In accordance with yet another embodiment, the peptide may include the following structure:
(Y33)a - X33 - (Z33)b' wherein Y33 and Z33 are as Previously defined, a is 0 or 1, and b is 0 or 1.
In accordance with yet another embodiment, when the peptide includes the structure X34, the peptide may include the following structure:
Y34 - X34. wherein X34 is as hereinabove described, and Y34 is:
(i) R32;
(ii) R32-R32; (iii) R31-R32-R32;
(iv) R32-R31-R32-R32;
(v) R32-R32-R31-R32-R32; or
(vi) R31-R32-R32-R31-R32-R32, wherein R31, R32 and R33 are as hereinabove described.
In accordance with another embodiment, when the peptide includes the structure X34, the peptide may include the following structure:
X34-Z34, wherein X34 is as hereinabove described, and Z34 is:
( (i) R33;
(ii) R33-R31;
(iii) R33-R31-R32;
( iv ) R33-R31-R32-R32 ;
( v ) R33- R31-R32-R32-R31 ; or
(vi) R33-R31-R32-R32-R31-R32.
In accordance with yet another embodiment, the peptide may include the following structure:
(Y34)a- X34- (Z34 ) b ' wherein X34 and Z34 are as previously defined, a is 0 or 1, and b is 0 or 1.
In accordance with a further embodiment, when the peptide includes the structure X35, the peptide may include the following structure:
Y35-X35, wherein X35 is as hereinabove described, and Y35 is:
(i) R33;
(ii ) R32-R33;
(iii) R32-R32-R33;
(iv) R31-R32-R32-R33;
(v) R32-R31-R32-R32-R33; or
(vi) R32-R32-R31-R32-R32-R33, wherein R31, R32, and R33 are hereinabove described.
In accordance with another embodiment, when the peptide includes the structure X35, the peptide may include the following structure:
X35 - Z35 wherein X35 is as hereinabove described, and Z35 is:
(i) R31;
(ii) R31-R32; (iii) R31-R32-R32;
(iv) R31-R32-R32-R31;
(v) R31-R32-R32-R31-R32; or
(vi) R31-R32-R32-R31-R32-R32.
In accordance with yet another embodiment, the peptide may include the following structure:
(Y35)a- X35 (Z35)b, wherein X35 and Z35 are as Previously defined, a is 0 or 1, and b is 0 or 1.
In accordance with a further embodiment, when the peptide includes the structure X36. the peptide may include the following structure:
Y36 - X36 wherein X36 is as hereinabove described, and Y36 is:
(i) R31;
(ii) R33-R31;
(iii) R32-R33-R31;
(iv) R32-R32-R33-R31;
(v) R31-R32-R32-R33-R31; or
(vi) R32-R31-R32-R32-R33-R31, wherein R31, R32, and R33 are as hereinabove described.
In accordance with another embodiment, when the peptide included the structure X36, the peptide may include the following structure:
X36-Z36, wherein X36 is as hereinabove described, and Z36 is:
(i) R32;
(ii) R32-R32;
(iii) R32-R32-R31;
(iv) R32-R32-R31-R32;
(v) R32-R32-R31-R32-R32; or
(vi) R32-R32-R31-R32-R32-R33.
In accordance with yet another embodiment, the peptide may include the following structure:
(Y36)a- X36 (Z36)b, wherein Y36 and Z36 are as previously defined, a is 0 or 1, and b is 0 or 1. In accordance with one embodiment, when the peptide includes the structure X37, the peptide may includes the structure Y37-X37, wherein X37 is as hereinabove described, and Y37 is:
(i) R32;
(ii} R31-R32;
(iii) R33-R31-R32;
(iv) R32-R33-R31-R32;
(v) R32-R32-R33-R31-R32; or
(vi) R31-R32-R32-R33-R31-R32, wherein R31, R32, and R33 are as hereinabove described.
In accordance with a further embodiment, when the peptide includes the structure X37, the peptide may include the following structure:
X37 - Z37 wherein X37 is as hereinabove described, and Z37 is: (i) R32;
(ii) R32-R31;
(iii) R32-R31-R32;
(iv) R32-R31-R32-R32;
(V) R32-R31R32-R32-R33; or
(vi) R32-R31-R32-R32-R33-R31.
In accordance with yet another embodiment, the peptide may include the following structure:
(Y37)a- X37 (Z37)b, wherein Y37 and Z37 are as previously defined, a is 0 or 1, and b is 0 or 1.
In a preferred embodiment, n is 3, and most preferably the peptide is of one of the following structures as given in the
accompanying sequence listing:
(Lys lle Ala Gly Lys lle Ala)3 (SEQ ID NO:27).
(Lys lle Ala Lys lle Ala Gly)3 (SEQ ID NO:28).
(Lys lle Ala Gly Lys lle Gly)3 (SEQ ID NO:29).
(Lys Leu Ala Gly Lys Leu Ala)3 (SEQ ID NO:30).
(Lys Phe Ala Gly Lys Phe Ala)3 (SEQ ID NO:31).
(Lys Ala Leu Ser Lys Ala Leu)3 (SEQ ID NO:32).
(Lys Leu Leu Lys Ala Leu Gly)3 (SEQ ID NO: 33). (Lys Ala lle Gly Lys Ala lle)3 (SEQ ID NO: 34).
(Gly lle Ala Lys lle Ala Lys)3 (SEQ ID NO: 35).
(Lys lle Ala Lys lle Phe Gly)3 (SEQ ID NO: 36).
(Gly lle Ala Arg lle Ala Lys)3 (SEQ ID NO: 37).
(Lys Phe Ala Arg lle Ala Gly)3 (SEQ ID NO:38).
(Gly Phe Ala Lys lle Ala Lys)3 (SEQ ID NO:39).
(Lys lle Ala Gly Orn lle Ala)3 (SEQ ID NO: 40).
(Lys lle Ala Arg lle Ala Gly)3 (SEQ ID NO: 41).
(Orn lle Ala Gly Lys lle Ala)3 (SEQ ID NO: 42).
(Gly lle Ala Arg lle Phe Lys)3 (SEQ ID NO: 43).
(Lys Nle Ala Gly Lys Nle Ala)3 (SEQ ID NO: 44).
(Lys Nle Ala Gly Lys lle Ala)3 (SEQ ID NO: 45).
(Lys lle Ala Gly Lys Nle Ala)3 (SEQ ID NO:46).
(Lys Nva Ala Gly Lys Nva Ala) 3 (SEQ ID NO: 47).
(Lys Nva Ala Gly Lys lle Ala)3 (SEQ ID NO: 48).
(Lys Leu Leu Ser Lys Leu Gly)3 (SEQ ID NO: 49).
(Lys Leu Leu Ser Lys Phe Gly)3 (SEQ ID NO: 50).
(Lys lle Ala Gly Lys Nva Ala)3 (SEQ ID NO: 51).
(His lle Ala Gly His lle Ala)3 (SEQ ID NO: 52).
(Ala Gly Lys lle Ala Lys lle)3 (SEQ ID NO: 53).
(lle Ala Lys lle Ala Gly Lys)3 (SEQ ID NO: 54).
(Lys lle Ala Gly Arg lle Ala)3 (SEQ ID NO: 55).
(Arg lle Ala Gly Arg lle Ala)3 (SEQ ID NO: 56).
(Lys Val Ala Gly Lys lle Ala)3 (SEQ ID NO: 57).
(Lys lle Ala Gly Lys Val Ala)3 (SEQ ID NO: 58).
(Ala Lys lle Ala Gly Lys lle) ID NO: 59).
3 (SEQ
(Orn lle Ala Gly Orn lle Ala)3 (SEQ ID NO: 60).
(Lys Phe Ala Gly Lys lle Ala)3 (SEQ ID NO: 61).
(Lys lle Ala Gly Lys Phe Ala)3 (SEQ ID NO: 62).
(Lys Cha Ala Gly Lys lle Ala)3 (SEQ ID NO: 63).
(Lys Nle Ala Lys lle Ala Gly)3 (SEQ ID NO: 64).
(Arg lle Ala Gly Lys lle Ala)3 (SEQ ID NO: 65).
(Har lle Ala Gly Har lle Ala)3 (SEQ ID NO: 66).
(Xaa lle Ala Gly Lys lle Ala)3 (SEQ ID NO: 67). (Lys lle Ala Gly Xaa lle Ala)3 (SEQ ID NO:68).
Lys lle Ala (Lys lle Ala Gly Lys lle Ala)3 (SEQ ID NO: 69)
In (SEQ ID NO: 67) and (SEQ ID NO: 68), Xaa is p-aminophenylalanme.
In accordance with another embodiment, the amphiphilic peptide includes the following basic structure X40:
R31-R32-R32-R33-R34-R32-R32-R31-R32-R32-R32-R34-R32-R32,
wherein R31, R32, and R33 are as hereinabove described, and R34 is a basic hydrophilic or hydrophobic amino acid.
In accordance with one embodiment, the peptide may include the following structure:
Y40-X40' wherein X40 is as hereinabove described, and Y40 is:
(i) R32;
(ii) R32-R32;
(iii) R34-R32-R32;
(iv) R33-R34-R32-R32;
(v) R32-R33-R34-R32-R32;
(v) R32-R32-R33-R34-R32-R32, or
( vii ) R31-R32-R32-R33-R34-R32-R32 , Wherein R31' R32 , R33 and R34 are as hereinabove described.
In accordance with another embodiment, the peptide may include the following structure:
X40-Z40, wherein X40 is as hereinabove-described and Z40 is:
(i) R31;
(ii) R31-R32;
(iii) R31-R32-R32;
(iv) R31-R32-R32-R33;
(v) R31-R32-R32-R33-R34;
(vi) R31-R32-R32-R33-R34-R32, or
(vii) R31-R32-R32-R33-R34-R32-R32, wherein R31, R32, R33, andR34 are as hereinabove described.
In accordance with yet another embodiment the peptide may include the following structure:
(Y40)a-X40-(Z40)b, wherein Y40 and Z40 are as previously defined, a is 0 or 1, and b is 0 or 1. In a preferred embodiment, the peptide has the following structural formula as given in the accompanying sequence listing:
(SEQ ID NO: 70)
In another preferred embodiment, the peptide has the following structural formula as given in the accompanying sequence l i sting :
(SEQ ID NO: 71)
In accordance with a further embodiment, the peptide has one of the one of the following structural formulae as given in the
accompanying sequence listing:
(SEQ ID NO: 72)
(SEQ ID NO: 73)
(SEQ ID NO: 74)
(SEQ ID NO: 75)
(SEQ ID NO: 76)
(SEQ ID NO: 77)
(SEQ ID NO: 78)
(SEQ ID NO: 79)
(SEQ ID NO: 80)
(SEQ ID NO: 81)
(SEQ ID NO: 82)
(SEQ ID NO: 83)
(SEQ ID NO: 84)
(SEQ ID NO: 85)
(SEQ ID NO:86)
(SEQ ID NO: 87)
In accordance with another embodiment, the peptide may include the following structural formula:
- (Lys lle Ala Lys Lys lle Ala)-n, wherein n is from 2 to 5. Preferably, n is 3, and the peptide has the following structural formula:
(Lys lle Ala Lys Lys lle Ala)3
(SEQ ID NO: 88)
In accordance with another embodiment, the peptide may include the following structural formula: -(Lys Phe Ala Lys Lys Phe Ala)-n
wherein n is from 2 to 5.
Preferably, n is 3, and the peptide has the following structural formula:
(Lys Phe Ala Lys Lys Phe Ala)3
(SEQ ID NO: 89)
In accordance with another embodiment, the peptide may includ the following structural formula:
-(Lys Phe Ala Lys Lys lle Ala)-n
wherein n is from 2 to 5. Preferably n is 3, and the peptide has the following structural formula:
(Lys Phe Ala Lys Lys lle Ala)3
(SEQ ID NO: 90).
In accordance with another embodiment, the peptide may be selected from the group consisting of the following structural formulae as given in the accompanying sequence listing:
(SEQ ID NO: 91)
(SEQ ID NO:92)
(SEQ ID NO: 93)
(SEQ ID NO: 94)
In accordance with yet another embodiment, the peptide may be a cecropin or sarcotoxin.
The term cecropins includes the basic structure as well as analogues and derivatives thereof. The cecropins and analogues and derivatives thereof are described in Ann. Rev. Microbiol. 1987, Vol. 41, pages 103-26, in particular page 108, and in Christensen, et al., PNAS Vol. 85, pgs. 5072-76, which are hereby incorporated by
reference.
The term sarcotoxins includes the basic materials as well as analogues and derivatives thereof. The sarcotoxins and analogues and derivatives thereof are described in Molecular Entomology, pages 369-78, in particular page 375, Alan R. Liss, Inc. (1987), which is hereby incorporated by reference. In another embodiment, the amphiphilic peptide includes the following basic structure X50 :
R41-R42-R42-R41-R42-R42-R41-R41-R42-R41-R41.
R41 is a hydrophobic amino acid, and R42 is a basic hydrophilic or neutral hydrophilic amino acid.
In one embodiment, the peptide includes the basic structure
Y50 -X50 wherein X50 is as hereinabove described and Y50 is:
(i) R41 ;
(ii) R42-R41: or
(iii) R42-R42-R41, wherein R41 and R42 are as hereinabove
described.
In one embodiment, R41 is leucine. In another embodiment, R42 is lysine. Representative examples of peptides in accordance with this aspect of the present invention include those having the following structures:
(SEQ ID NO 95)
(SEQ ID NO 96)
(SEQ ID NO 97)
(SEQ ID NO 98)
In accordance with another embodiment, the amphiphilic peptide includes the following basic structure X50 :
R42-R41-R42-R42-R41-R41-R42-R42-R41-R42-R42, wherein R41 is a hydrophobic amino acid and R42 is a basic hydrophilic or neutral hydrophilic amino acid.
In one embodiment R4 1 is leucine. In another embodiment, R42 is lysine.
In one embodiment, the peptide includes the basic structure
wherein X52 is as hereinabove described, and Y52 is:
Y52- X52 , Wherein X52 is as hereinabove described, and Y52 is:
( i ) R42;
( ii) R41-R42;
( iii ) R41-R41-R42;
( iv) R42-R41-R41-R42; or
(v) R42-R42-R41-R41-R42. In one embodiment, the peptide may have the following structure; Lys Lys Leu Leu Lys Lys Leu Lys Lys Leu
5 10
Leu Lys Lys Leu Arg Arg
15
(SEQ ID NO: 99)
In another embodiment, the peptide includes the basic structure
X52 - Z52, wherein X52 is as hereinabove described, and Z52 is:
(i) R41;
(ii) R41-R41;
( i i i ) R41-R41- R42 ;
(iv) R41-R41-R42-R42; or
(v) R41-R41-R42-R42-R41;
In one embodiment, the peptide may have the following structure:
Lys Leu Lys Lys Leu Leu Lys Lys Leu Lys Lys Leu Leu Lys Lys Leu
5 10 15
(SEQ ID NO: 100)
In another embodiment, the peptide may include the structure:
(Y52)a - X52 - (Z52)b' wherein X52, Y52 and Z52 are as
hereinabove described, and a is 0 or 1, and b is 0 or 1.
In accordance with another embodiment, the peptide includes the following basic structure X54:
-R41-R42-R42-R41-R41-R42-R42-R41-R42-R42-R41-R41-R42-R42-R43-, wherein R41 and R42 are as hereinabove described, and R43 is a netural hydrophilic amino acid.
In one embodiment, the peptide may have the following structure:
(SEQ ID NO: 101)
In another embodiment, the peptide may have the following
structure:
(SEQ ID NO: 102)
In accordance with yet another embodiment, the peptide includes the following basic structure X56: R41-R42-R41-R41-R42-R42-R41-R41-R42-R42-R44, wherein R41 and R42 are as hereinabove described, and R44 is a neutral hydrophilic amino acid or proline.
In one embodiment, the peptide may include the following
structure Y56-X56' wherein X56 is the basic peptide structure
hereinabove described, and Y56 is:
(i) -R41
(i) -R41-R41;
( i i i ) -R42-R41-R41 ;
(iv) -R41-R42-R41-R41;
(v) -R41-R41-R42-R41-R41;
(vi) -R42-R41-R41-R42-R41-R41; or
(vii) -R42-R42-R41-R41-R41-R42-R41-R41,
wherein R41 and R42 are as hereinabove described.
In one embodiment, the peptide may include the structure:
X56-Z56' wherein X56 is as hereinabove described, and Z56 is:
(i) -R 42;
(ii ) -R42-R42;
(iii) -R42-R42-R41;
(iv) -R42-R42-R41-R41;
(v) -R42-R42-R41-R41-R42;
(vi) -R42-R42-R41-R41-R42-R42; or
(vii) -R42-R42-R41-R41-R42-R42-R41.
In a preferred embodiment, the peptide may have one of the following structures:
(SEQ ID NO: 103); or
(SEQ ID NO: 104).
In another embodiment, the peptide may have the structure
(Y56)a-X 56-(Z56)b, wherein X56, Y56, and Z56 are as hereinabove described, a is 0 or 1, and b is 0 or 1.
In accordance with another embodiment, the peptide includes the following basic structure X58:
R41-R41-R42-R42-R41-R42-R42-R41-R41-R42-R42-R41-R43,
wherein R41, R42 and R43 are as hereinabove described. In accordance with another embodiment, the peptide may include the structure Y58-X58, wherein X58 is as hereinabove described, and
Y58 is:
(i) -R41;
(ii ) -R42-R41;
( i i i ) -R42-R42-R41 ;
(iv) -R41-R42-R42-R41;
(v) -R41-R41-R42-R42-R41;
(vi) -R42-R41-R41-R42-R42-R41; or
(vii) -R42-R42-R41-R41-R42-R42-R41,wherein R41 and R42 are as hereinabove described.
In another embodiment, the peptide includes the structure X58-Z58, wherein X58 is as hereinabove described, and Z58 is:
(i) -R41;
(ii ) -R41-R45;
(iii) -R41-R45-R45;
(iv) -R41-R45-R45-R43;
(v) -R41-R45-R45-R43-R41;
(vi) -R41-R45-R45-R43-R41-R43:
(vii) -R41-R45-R45-R43-R41-R43-R43,
(viii) -R41-R45-R45-R43-R41-R43-R43-R45; or
(ix) -R41-R45-R45-R43-R41-R43-R43-R45-R43, wherein R41 and R43 are as hereinabove described, and R45 is proline.
In one embodiment, the peptide has the following structure:
(SEQ ID NO: 105).
In one embodiment, the peptide may have the structure
(Y58)a- X58-(Z58)b, wherein X58, Y58, and Z58 are as hereinabove described, a is 0 or 1, and b is 0 or 1.
In accordance with another embodiment, the peptide includes the following basic structure X60;
R41-R41-R43-R42-R41-R41-R41-R41-R41-R41-R42-R41-R41-R42-R42-R41- R41-R42-R42-R42-R41' wherein R41, R42, and R43 are as hereinabove described. In one embodiment, the peptide may have the following structure:
(SEQ ID NO: 106).
In another embodiment, the peptide may include the structure X60 -Z60, wherein X60 is as hereinabove described, and Z60 is:
(i) -R42;
( ii ) -R42-R42 ,
( iii ) -R42-R42-R41 ;
(iv) -R42-R42-R41-R41;
(v) -R42-R42-R41-R41-R42;
(vi) -R42-R42-R41-R41-R42-R42; or
(vii) -R42-R42-R41-R41-R42-R42-R41.
In accordance with yet another embodiment, the peptide has a structure selected from the group consisting of:
(a) R41-R42-R42-R41-R42-R42-R41;
(b) R41-R41-R42-R42-R41-R42-R42-R41;
(c) R42-R41-R41-R42-R42-R41-R42-R42-R41;
( d ) R42-R42-R41-R41-R42-R42-R41-R42-R42-R41 ; and
(e) R41-R42-R42-R41-R41-R42-R42-R41-R42-R42-R41, wherein R41 and R42 are as hereinabove described.
In one embodiment, the peptide has the structure (a), and a representative example of such a structure is (SEQ ID NO: 107), which is given in the accompanying sequence listing.
In another embodiment, the peptide has the structure (b), and representative example of such a structure is (SEQ ID NO: 108), which is given in the accompanying sequence listing.
In another embodiment, the peptide has the structure (c), and a representative example of such a structure is (SEQ ID NO: 109) as given the accompanying sequence listing.
In yet another embodiment, the peptide has the structure (d), and a representative example of such a structure is (SEQ ID NO: 110) as given in the accompanying sequence listing. In a further embodiment, the peptide has the structure (e), and representative examples of such a structure are (SEQ ID NO: 111) and (SEQ ID NO: 112) as given in the accompanying sequence listing.
In accordance with another embodiment, the peptide has the following structural formula:
(SEQ ID NO: 113).
In accordance with another embodiment, the peptide is melittin.
Melittin is an amphipathic peptide consisting of 26 amino acid residues, and is isolated from honeybee (Apis mellifera) venom. The peptide is known to be cytolytic. See Habermann, et al.,
Hoppe-Seyler's Zeitschrift Physiol. Chem., Vol. 348, pgs. 37-50 (1987). Melittin has the following structural formula as represented by the three-letter amino acid code:
Gly lle Gly Ala Val Leu Lys Val Leu
5
Thr Thr Gly Leu Pro Ala Leu lle Ser
10 15
Trp lle Lys Arg Lys Arg Gln Gln
20 25
(SEQ ID NO: 114)
In another embodiment, the peptide purified in accordance with the present invention is an apidaecin. The term apidaecin as used herein includes the basic structure as well as analogues and
derivaties thereof. Apidaecins are further described in European Patent Application No. 299,828.
In accordance with another embodiment, the peptide may be an amide - or carboxy-terminated peptide represented by the following structural formula, and the numbers below each amino acid residue refer to the position of the residue in the peptide:
Leu Lys Leu Leu Lys Lys Leu Leu Lys Lys Leu
1 2 3 4 5 6 7 8 9 10 11
Lys Lys Leu Leu Lys Lys Leu
12 13 14 15 16 17 18 (SEQ ID NO: 115)
or the peptide may be an analogue of such peptide wherein at least one of amino acid residues 1 through 7, 9, 11, 12, 14, 16, or 18 is deleted from the peptide.
In one embodiment, at least one of amino acid residues 1, 3, 7, 9, 11, 12, 14, 16, or 18 is deleted from the peptide. In other embodiments, amino acid residues 1 through 3, 1 through 4, 1 through 5, 1 through 6, and 1 through 7 are deleted from the peptide.
In preferred embodiments, amino acid residues 1 through 3 or 1 through 4 are deleted from the peptide, and such peptides have the following structural formulae:
(SEQ ID NO: 116)
(SEQ ID NO: 117)
In accordance with another embodiment, the peptide includes the following structural formula X62:
R41-R41-R42-R41-R41-R42-R42-R41-R41-R42-R42-
R41-R42-R42,
wherein R41 is a hydrophobic amino acid, and R42 is a basic
hydrophilic or neutral hydrophilic amino acid. In one embodiment, R41 is leucine, and in another embodiment, R42 is lysine. In a preferred embodiment, the peptide has the following structure:
Leu Leu Lys Leu Leu Lys Lys Leu Leu Lys Lys Leu Lys Lys
5 10
(SEQ ID NO: 118)
In accordance with another embodiment, the peptide includes the following structural formula X64:
R42-R41-R41-R42-R42-R41-R42-R42-R41-R41-R42-R42
-R41-R41,
wherein R41 is a hydrophobic amino acid, and R42 is a basic
hydrophilic or neutral hydrophilic amino acid. In one embodiment, R41 is leucine, and in another embodiment, R42 is lysine. In a preferred embodiment, the peptide has the following structural formula:
Lys Leu Leu Lys Lys Leu Lys Lys Leu Leu Lys Lys Leu Leu
5 10
(SEQ ID NO: 119)
In another embodiment, the peptide includes the following
structural formula X66 :
R41-R41-R42-R42-R41-R42-R42-R41-R41-R42-R42-R41-R41,
wherein R41 is a hydrophobic amino acid, and R42 is a basic
hydrophilic or neutral hydrophilic amino acid.
In one embodiment, the peptide may include the following
structure:
X66-Z66, wherein X66 is as hereinabove described and Z66 is:
(i) R42;
(ii ) R42-R41; or
(iii) R42-R41-R41
In one embodiment, R41 is leucine, and in another embodiment, R42 is lysine. In a preferred embodiment, the peptide has the following structural formula:
Leu Leu Lys Lys Leu Lys Lys Leu Leu Lys Lys Leu Leu Lys Leu Leu
5 10 15
(SEQ ID NO: 120)
In accordance with yet another embodiment, the amphiphilic peptide or protein may be an ion channel-forming peptide or protein.
Ion channel-forming proteins or peptides which may be employed include defensins, also known as human neutrophil antimicrobial peptides (HNP), major basic protein (MBP) of eosinophils, bactericidal permeability-increasing protein (BPT). 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 (1985). 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. 14891-14894 (1987). Perforin is described in Henkart, et al., J. Exp. Med., 160 75 (1984), and in Podack, et al., J. Exp. Med., 160:695 (1984). The above articles are hereby incorporated by reference.
The term ion channel-forming proteins includes the basic
structures of the ion channel-forming proteins as well as analogues and derivatives.
The peptides may be produced by known techniques and obtained in substantially pure form. For example, the peptides may be synthesized on an automatic peptide 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 codons encoding the amino acids are known to those skilled in the art, and thus one may construct DNA encoding any of the peptides by accepted techniques, and clone such DNA into an expression vehicle such as, for example, a plasmid, and transfect such an expression vehicle into a cell which will express the peptide.
The peptide or protein and protease inhibitor may be
administered in combination with chemotherapeutic agents such as, but not limited to, cyclophosphamide, cisplatin, doxorubicin,
hexamethylamine, and VP-16.
The peptide or protein and protease inhibitor may be administered before, during, or after radiation treatment, chemotherapy, or
surgery. The administration of the peptides or proteins and protease inhibitor during surgery or radiation treatment may be advantageous in inhibiting, preventing, and/or destroying potential "loose" malignant cells capable of colonizing other sites.
The invention will now be described with respect to the following example; however, the scope of the present invention is not intended to be limited thereby.
Example The following melanoma cel l lines are seeded in 200 ul of 2 . 0% tumor medium at a concentration of 1.5 x 104 cells per well in a
96-well plate:
WM 1158
WM 793
WM 1366
WM 902B
After 1 day, the media are removed, and peptide D-(SEQ ID
NO: 117), wherein each amino acid residue is a D-amino acid residue, is added to one group of each of the cell lines in triplicate in a concentration of 1.0 ug/ml. To another group of each of the cell lines, bestatin is added in triplicate at a concentration in 2% medium of 70 ug/ml. To another group of the cell lines, D-(SEQ ID NO: 117) is added at a concentration of 1.0 ug/ml and bestatin is added at a concentration 70 ug/ml in 2% medium in triplicate. A control group has neither peptide nor bestatin added to the cells.
On the next day, 10 ul/lμCi/ well of 3H-thymidine i s added to each well with a multichannel pipetter. After 18 hours, the medium is discarded as radioactive waste, and 50 μl of trypsin/versene solution is added to each well and placed into an incubator until detached.
Detached cells are harvested on a Tomtec cell harvester, and
radioactive counts are measured from the filters on a Packard counter. The radioactive counts for each cell line contacted with the peptides is given in Table I below.
Figure imgf000035_0001
% decrease is based upon the difference between the results with bestatin only and the results with D-(SEQ ID NO: 117) and bestatin.
The peptide or protein and protease inhibitor, as hereinabove described, may be employed for treating a wide variety of hosts. The host may be a human or non-human animal. The peptide or protein and the protease inhibitor may be employed together in a single
composition, or in separate compositions. Moreover, the protease inhibitor and the peptide or protein may be delivered or administered in different forms.
The peptide or protein and protease inhibitor 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. 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 or protein and protease inhibitor may also be used in combination with adjuvants, or compatible drugs where such a combination is seen to be desirable or advantageous in treating a cancerous growth.
The peptide(s) or protein of the present invention may be administered to a host; in particular an animal, in an effective amount for treating a cancerous growth in conjunction with a protease inhibitor for potentiating the activity of the peptide or protein.
Numerous modifications and variations of the present invention are possible in light of the above teachings and, therefore, within the scope of the appended claims, the invention may be practiced otherwise than as particularly described.
SEQUENCE LISTING
(1) GENERAL INFORMATION:
(i) APPLICANT: Herlyn, Meenhard
Jacob, Leonard S.
Maloy, W. Lee
(ii) TITLE OF INVENTION: Treatment of Cancerous Growths with
Biologically Active Peptides and
Protease Inhibitors
(iii) NUMBER OF SEQUENCES: 120 (iv) CORRESPONDENCE ADDRESS:
(A) ADDRESSEE: Carella, Byrne, Bain, Gilfillan,
Cecchi & Stewart
(B) STREET: 6 Becker Farm Road
(C) CITY: Roseland
(D) STATE: New Jersey
(E) COUNTRY: USA
(F) ZIP: 07068
(v) COMPUTER READABLE FORM:
(A) MEDIUM TYPE: 3.5 inch diskette
(B) COMPUTER: IBM PS/2
(C) OPERATING SYSTEM: PC-DOS
(D) SOFTWARE: DW4.V2
(vi) CURRENT APPLICATION DATA:
(A) APPLICATION NUMBER:
(B) FILING DATE:
(C) CLASSIFICATION: (vii) PRIOR APPLICATION DATA:
(A) APPLICATION NUMBER:
(B) FILING DATE:
(viii) ATTORNEY/AGENT INFORMATION:
(A) NAME: Olstein, Elliot M.
(B) REGISTRATION NUMBER: 24,025
(C) REFERENCE/DOCKET NUMBER: 421250
(ix) TELECOMMUNICATION INFORMATION:
(A) TELEPHONE: 201-994-1700
(B) TELEFAX: 201-994-1744
(2) INFORMATION FOR SEQ ID NO:1:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 20 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(x) PUBLICATION INFORMATION:
(H) DOCUMENT NUMBER: W089/11290
(I) FILING DATE: 19-MAY-1989
(J) PUBLICATION DATE: 30-NOV-1989
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:1:
Ala Phe Ser Lys Ala Phe Ser Lys Ala Phe
5 10
Ser Lys Ala Phe Ser Lys Ala Phe Ser Lys
15 20 ( 2 ) INFORMATION FOR SEQ ID NO : 2 :
( i ) SEQUENCE CHARACTERI STICS :
( A ) LENGTH : 24 amino acids
( B ) TYPE : amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(x) PUBLICATION INFORMATION:
(H) DOCUMENT NUMBER: W089/11290
(I) FILING DATE: 19-MAY-1989
(J) PUBLICATION DATE: 30-NOV-1989
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 2: Ala Phe Ser Lys Ala Phe Ser Lys Ala Phe
5 10
Ser Lys Ala Phe Ser Lys Ala Phe Ser Lys
15 20
Ala Phe Ser Lys
(2) INFORMATION FOR SEQ ID NO: 3:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 16 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(x) PUBLICATION INFORMATION:
(H) DOCUMENT NUMBER: W089/11290 (I) FILING DATE: 19-MAY-1989
(J) PUBLICATION DATE: 30-NOV-1989 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3: Phe Ser Lys Ala Phe Ser Lys Ala Phe Ser
5 10
Lys Ala Phe Ser Lys Ala
15
(2) INFORMATION FOR SEQ ID NO: 4:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 20 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(x) PUBLICATION INFORMATION:
(H) DOCUMENT NUMBER: -W089/11290
(I) FILING DATE: 19-MAY-1989
(J) PUBLICATION DATE: 30-NOV-1989
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:4: Ser Lys Ala Phe Ser Lys Ala Phe Ser Lys
5 10
Ala Phe Ser Lys Ala Phe Ser Lys Ala Phe
15 20
(2) INFORMATION FOR SEQ ID NO: 5:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 16 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide (x) PUBLICATION INFORMATION:
(H) DOCUMENT NUMBER: W089/11290
(I) FILING DATE: 19-MAY-1989
(J) PUBLICATION DATE: 30-NOV-1989
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:5: Lys Ala Phe Ser Lys Ala Phe Ser Lys Ala
5 10
Phe Ser Lys Ala Phe Ser
15
(2) INFORMATION FOR SEQ ID NO: 6:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 23 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(A) NAME/KEY: Magainin I peptide.
(x) PUBLICATION INFORMATION:
(A) AUTHOR: Zasloff, Michael
(C) JOURNAL: Proc . Nat. Acad. Sci.
(D) VOLUME: 84
(F) PAGES: 5449-5453
(G) DATE: AUG - 1987
( H ) DOCUMENT NUMBER : US 4810777
( I ) FI LING DATE : 04-MAR- 1987
( J ) PUBLICATION DATE : 07 -MAR- 1989 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:6: Gly lle Gly Lys Phe Leu His Ser Ala Gly
5 10
Lys Phe Gly Lys Ala Phe Val Gly Glu lle
15 20
Met Lys Ser
(2) INFORMATION FOR SEQ ID NO: 7:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 23 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(A) NAME/KEY: Magainin II peptide.
(x) PUBLICATION INFORMATION:
(A) AUTHOR: Zasloff, Michael
(C) JOURNAL: Proc. Nat. Acad. Sci.
(D) VOLUME: 84
(F) PAGES: 5449-5453
(G) DATE: AUG - 1987
(H) DOCUMENT NUMBER: US 4810777
(I) FILING DATE: 04-MAR-1987
(J) PUBLICATION DATE: 07-MAR-1989
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:7: Gly lle Gly Lys Phe Leu His Ser Ala Lys
5 10
Lys Phe Gly Lys Ala Phe Val Gly Glu lle
15 20
Met Asn Ser
(2) INFORMATION FOR SEQ ID NO: 8:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 22 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(A) NAME/KEY: Magainin III peptide.
(x) PUBLICATION INFORMATION:
(A) AUTHOR: Zasloff, Michael
(C) JOURNAL: Proc. Nat. Acad. Sci.
(D) VOLUME: 84
(F) PAGES: 5449-5453
(G) DATE: AUG - 1987
(H) DOCUMENT NUMBER: US 4810777
(I) FILING DATE: 04-MAR-1987
(J) PUBLICATION DATE: 07-MAR-1989
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:8:
Gly lle Gly Lys Phe Leu His Ser Ala Lys
5 10
Lys Phe Gly Lys Ala Phe Val Gly Glu Ile
15 20
Met Asn
(2) INFORMATION FOR SEQ ID NO: 9:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 22 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(A) NAME/KEY: magainin peptide.
(x) PUBLICATION INFORMATION:
(A) AUTHOR: Zasloff, Michael
(C) JOURNAL: Proc. Nat. Acad. Sci.
(D) VOLUME: 84
(F) PAGES: 5449-5453
(G) DATE: AUG - 1987
(H) DOCUMENT NUMBER: US 4810777
(I) FILING DATE: 04-MAR-1987
(J) PUBLICATION DATE: 07-MAR-1989
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:9: lle Gly Lys Phe Leu His Ser Ala Lys Lys
5 10
Phe Gly Lys Ala Phe Val Gly Glu lle Met
15 20
Asn Ser
(2) INFORMATION FOR SEQ ID NO: 10:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(A) NAME/KEY: magainin peptide.
(x) PUBLICATION INFORMATION:
(A) AUTHOR: Zasloff, Michael
(C) JOURNAL: Proc. Nat. Acad. Sci.
(D) VOLUME: 84
(F) PAGES: 5449-5453
(G) DATE: AUG - 1987
(H) DOCUMENT NUMBER: US 4810777
(I) FILING DATE: 04-MAR-1987
(J) PUBLICATION DATE: 07-MAR-1989
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:10:
Gly Lys Phe Leu His Ser Ala Lys Lys Phe
5 10
Gly Lys Ala Phe Val Gly Glu lle Met Asn
15 20
Ser
(2) INFORMATION FOR SEQ ID NO:11:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 20 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(A) NAME/KEY: magainin peptide.
(x) PUBLICATION INFORMATION:
(A) AUTHOR: Zasloff, Michael
(C) JOURNAL: Proc. Nat. Acad. Sci.
(D) VOLUME: 84
(F) PAGES: 5449-5453
(G) DATE: AUG - 1987
(H) DOCUMENT NUMBER: US 4810777
(I) FILING DATE: 04-MAR-1987
(J) PUBLICATION DATE: 07-MAR-1989
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 11: Lys Phe Leu His Ser Ala Lys Lys Phe Gly
5 10
Lys Ala Phe Val Gly Glu lle Met Asn Ser
15 20 (2) INFORMATION FOR SEQ ID NO:12:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(A) NAME/KEY: PGLa peptide.
(x) PUBLICATION INFORMATION:
(A) AUTHOR: Hoffman, et al.
(C) JOURNAL : EMBO J .
(D ) VOLUME: 2
(F) PAGES: 711-714
(G) DATE: 1983
(A) AUTHOR: Andreu, et al.
(C) JOURNAL: Journal of Biochemistry
(D) VOLUME: 149
(F) PAGES: 531-535
(G) DATE: 1985
(A) AUTHOR: Gibson, et al.
(C) JOURNAL: J. Biol. Chem.
(D) VOLUME: 261
(F) PAGES: 5341-5349
(G) DATE: 1986
(A) AUTHOR: Giovannini, et al.
(C ) JOURNAL: Biochem J.
(D ) VOLUME: 243
(F ) PAGES: 113-120
(G ) DATE: 1987 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 12
Gly Met Ala Ser Lys Ala Gly Ala lle Ala
5 10
Gly Lys lle Ala Lys Val Ala Leu Lys Ala
15 20
Leu
(2) INFORMATION FOR SEQ ID NO:13:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 25 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(A) NAME/KEY: XPF peptide.
(x) PUBLICATION INFORMATION:
(A) AUTHOR: Hoffman, et al.1
(C) JOURNAL: EMBO J.
(D) VOLUME: 2
(F)) PAGES: 711-714
(G) DATE: 1983
(A) AUTHOR: Andreu, et al. (C) JOURNAL: Journal of Biochemistry (D) VOLUME: 149
(F) PAGES: 531-535
(G) DATE: 1985
(A) AUTHOR: Gibson, et al. (C) JOURNAL: J. Biol. Chem. (D) VOLUME: 261
(F) PAGES: 5341-5349 (G) DATE: 1986
(A) AUTHOR : Giovannini, et al.
(C) JOURNAL : Biochem J.
(D) VOLUME: 243
(F) PAGES: 113-120
(G) DATE: 1987
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 13: Gly Trp Ala Ser Lys lle Gly Gln Thr Leu
5 10
Gly Lys lle Ala Lys Val Gly Leu Lys Glu
15 20
Leu lle Gln Pro Lys
25
(2) INFORMATION FOR SEQ ID NO:14:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 27 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(A) NAME/KEY: CPF peptide.
(x) PUBLICATION INFORMATION:
(A) AUTHOR: Richer, K.
Egger, R.
Kreil
(C) JOURNAL: J. Biol. Chem.
(D) VOLUME: 261
(F) PAGES: 3676-3680 (G) DATE: 1986
(A) AUTHOR: Wakabayashi, T.
Kato, H.
Tachibaba, S.
(C) JOURNAL: Nucleic Acids Research
(D) VOLUME : 13
(F) PAGES : 1817-1828
(G) DATE: 1985
(A) AUTHOR: Gibson, B.W.
Poulter, L.
Williams, D.H.
Maggio, J.E.
(C) JOURNAL : J. Biol. Chem.
(D) VOLUME : 261
(F) PAGES : 5341-5349
(G) DATE: 1986
(H) DOCUMENT NUMBER: W090/04407
(I) FILING DATE: 16-OCT-1989
(J) PUBLICATION DATE: 03-MAY-1990
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:14:
Gly Phe Gly Ser Phe Leu Gly Leu Ala Leu
5 10
Lys Ala Ala Leu Lys lle Gly Ala Asn Ala
15 20
Leu Gly Gly Ala Pro Gln Gln
25
(2) INFORMATION FOR SEQ ID NO:15:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 27 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(A) NAME/KEY: CPF peptide.
(x) PUBLICATION INFORMATION:
(A) AUTHOR: Richter, K
Egger, R.
Kreil
(C) JOURNAL: J. Biol. Chem.
(D) VOLUME: 261
(F) PAGES: 3676-3680
(G) DATE: 1986
(A) AUTHOR: Wakabayashi, T.
Kato, H.
Tachibaba, S.
(C) JOURNAL: Nucleic Acids Research
(D) VOLUME : 13
(F) PAGES : 1817-1828
(G) DATE: 1985
(A) AUTHOR : Gibson, B.W.
Poulter, L.
Williams, D.H.
Maggio, J.E.
(C) JOURNAL: J. Biol. Chem.
(D) VOLUME: 261
(F) PAGES : 5341-5349
(G) DATE: 1986
(H) DOCUMENT NUMBER: W090/04407
( I ) FILING DATE: 16-0CT-1989
(J) PUBLICATION DATE: 03-MAY-1990 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:15:
Gly Leu Ala Ser Phe Leu Gly Lys Ala Leu
5 10
Lys Ala Gly Leu Lys lle Gly Ala His Leu
15 20
Leu Gly Gly Ala Pro Gln Gln
25
(2) INFORMATION FOR SEQ ID NO: 16:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 27 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(A) NAME/KEY: CPF peptide.
(x) PUBLICATION INFORMATION:
(A) AUTHOR: Richter. K.
Egger, R.
Kreil
(C) JOURNAL: J. Biol. Chem.
(D) VOLUME: 261
(F) PAGES: 3676-3680
(G) DATE: 1986
(A) AUTHOR: Wakabayashi, T.
Kato, H.
Tachibaba, S.
(C) JOURNAL: Nucleic Acids Research
(D) VOLUME: 13
(F) PAGES:. 1817-1828 (G) DATE: 1985
(A) AUTHOR: Gibson, B.W.
Poulter, L.
Williams, D.H.
Maggio, J.E.
(C) JOURNAL: J. Biol. Chem.
(D) VOLUME: 261
(F) PAGES: 5341-5349
( G ) DATE : 1986
(H) DOCUMENT NUMBER: W090/04407
(I) FILING DATE: 16-OCT-1989
(J) PUBLICATION DATE: 03-MAY-1990
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:16: Gly Leu Ala Ser Leu Leu Gly Lys Ala Leu
5 10
Lys Ala Gly Leu Lys lle Gly Thr His Phe
15 20
Leu Gly Gly Ala Pro Gln Gln
25
(2) INFORMATION FOR SEQ ID NO: 17:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 27 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
( A ) NAME/KEY : CPF peptide .
( x ) PUBLICATION INFORMATI ON : (A) AUTHOR: Richter, K.
Egger, R.
Kreil
(C) JOURNAL: J. Biol. Chem.
(D) VOLUME: 261
(F) PAGES: 3676-3680
(G) DATE: 1986
(A) AUTHOR: Wakabayashi, T.
Kato, H.
Tachibaba, S.
(C) JOURNAL: Nucleic Acids Research
(D) VOLUME: 13
(F) PAGES: 1817-1828
(G) DATE : 1985
(A) AUTHOR: Gibson, B.W.
Poulter, L.
Williams, D.H.
Maggio, J.E.
(C) JOURNAL: J. Biol. Chem.
(D) VOLUME: 261
(F) PAGES: 5341-5349
(G) DATE: 1986
(H) DOCUMENT NUMBER: W090/04407
(I) FILING DATE: 16-OCT-1989
(J) PUBLICATION DATE: 03-MAY-1990
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:17:
Gly Leu Ala Ser Leu Leu Gly Lys Ala Leu
5 10
Lys Ala Thr Leu Lys lle Gly Thr His Phe
15 20
Leu Gly Gly Ala Pro Gln Gln
25 (2) INFORMATION FOR SEQ ID NO:18:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 27 amino acids
(B) TYPE: amino acid
( C ) STRANDEDNESS :
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(A) NAME/KEY: CPF peptide.
(x) PUBLICATION INFORMATION:
(A) AUTHOR: Richter, K.
Egger, R.
Kreil
(C) JOURNAL: J. Biol. Chem.
(D) VOLUME: 261
(F) PAGES: 3676-3680
(G) DATE: 1986
(A) AUTHOR: Wakabayashi, T.
Kato, H.
Tachibaba, S.
(C) JOURNAL: Nucleic Acids Research
(D) VOLUME: 13
(F) PAHES: 1817-1828
(G) DATE: 1985
(A) AUTHOR: Gibson, B.W.
Poulter, L.
Williams, D.H.
Maggio, J.E.
(C) JOURNAL J. Biol. Chem.
(D) VOLUME 261
(F) PAGES: 5341-5349 ( G ) DATE : 1986
(H) DOCUMENT NUMBER: W090/04407
(I) FILING DATE: 16-OCT-1989
(J) PUBLICATION DATE: 03-MAY-1990
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:18: Gly Phe Ala Ser Phe Leu Gly Lys Ala Leu
5 10
Lys Ala Ala Leu Lys lle Gly Ala Asn Met
15 20
Leu Gly Gly Thr Pro Gln Gln
25
(2) INFORMATION FOR SEQ ID NO: 19:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 27 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(A) NAME/KEY: CPF peptide.
(x) PUBLICATION INFORMATION:
(A) AUTHOR: Richter, K.
Egger. R.
Kreil
(C) JOURNAL: J. Biol. Chem.
(D) VOLUME: 261
(F) PAGES: 3676-3680
(G) DATE: 1986
(A) AUTHOR: Wakabayashi, T. Kato, H.
Tachibaba, S.
(C) JOURNAL: Nucleic Acids Research
(D) VOLUME : 13
(F) PAGES : 1817-1828
(G) DATE: 1985
(A) AUTHOR: Gibson, B.W.
Poulter, L.
Williams, D.H.
Maggio, J.E.
(C) JOURNAL: J. Biol. Chem.
(D) VOLUME : 261
(F) PAGES: 5341-5349
(G) DATE: 1986
(H) DOCUMENT NUMBER: W090/04407
(I) FILING DATE: 16-OCT-1989
(J) PUBLICATION DATE: 03-MAY-1990
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:19: Gly Phe Gly Ser Phe Leu Gly Lys Ala Leu
5 10
Lys Ala Ala Leu Lys lle Gly Ala Asn Ala
15 20
Leu Gly Gly Ala Pro Gln Gln
25
(2) INFORMATION FOR SEQ ID NO: 20:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 27 ammo acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide (ix) FEATURE:
(A) NAME/KEY: CPF peptide,
(x) PUBLICATION INFORMATION:
(A) AUTHOR: Richter, K.
Egger, R.
Kreil
(C) JOURNAL: J. Biol. Chem.
(D) VOLUME: 261
(F) PAGES: 3676-3680
(G) DATE: 1986
(A) AUTHOR: Wakabayashi, T.
Kato, H.
Tachibaba, S.
(C) JOURNAL: Nucleic Acids Research
(D) VOLUME: 13
(F) PAGES: 1817-1828-
(G) DATE: 1985
(A) AUTHOR: Gibson, B.W.
Poulter, L.
Williams, D.H.
Maggio, J.E.
(C) JOURNAL: J. Biol. Chem.
(D) VOLUME: 261
(F) PAGES: 5341-5349
(G) DATE: 19B6
(H) DOCUMENT NUMBER: W090/04407
(I) FILING DATE: 16-OCT-1989
(J) PUBLICATION DATE: 03-MAY-1990
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:20: Gly Phe Gly Ser Phe Leu Gly Lys Ala Leu
5 10
Lys Ala Ala Leu Lys lle Gly Ala Asn Ala
15 20
Leu Gly Gly Ser Pro Gln Gln
25
(2) INFORMATION FOR SEQ ID NO: 21:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 27 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(A) NAME/KEY: CPF peptide.
(x) PUBLICATION INFORMATION:
(A) AUTHOR: Richter, K.
Egger, R.
Kreil
(C) JOURNAL: J. Biol. Chem.
(D) VOLUME: 261
(F) PAGES: 3676-3680
(G) DATE: 1986
(A) AUTHOR: Wakabayashi, T.
Kato, H.
Tachibaba, S.
(C) JOURNAL: Nucleic Acids Research
(D) VOLUME: 13
(F) PAGES: 1817-1828 (G) DATE: 1985
(A) AUTHOR: Gibson, B.W.
Poulter, L.
Williams, D.H.
Maggio, J.E.
(C) JOURNAL: J. Biol. Chem.
(D) VOLUME: 261
(F) PAGES: 5341-5349
(G) DATE: 1986
(H) DOCUMENT NUMBER: W090/04407
(I) FILING DATE: 16-OCT-1989
(J) PUBLICATION DATE: 03-MAY-1990
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 21: Gly Phe Ala Ser Phe Leu Gly Lys Ala Leu
5 10
Lys Ala Ala Leu Lys lle Gly Ala Asn Leu
15 20
Leu Gly Gly Thr Pro Gln Gln
25
(2) INFORMATION FOR SEQ ID NO:22:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 27 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(A) NAME/KEY: CPF peptide.
( x ) PUBLICATION INFORMATION: (A) AUTHOR: Richter, K.
Egger, R.
Kreil
(C) JOURNAL: J. Biol. Chem.
(D) VOLUME: 261
(F) PAGES: 3676-3680
(G) DATE: 1986
(A) AUTHOR: Wakabayashi, T.
Kato, H.
Tachibaba, S.
(C) JOURNAL: Nucleic Acids Research
(D) VOLUME: 13
(F) PAGES : 1817-1828
(G) DATE: 1985
(A) AUTHOR: Gibson, B.W.
Poulter, L.
Williams, D.H.
Maggio, J.E.
(C) JOURNAL: J. Biol. Chem.
(D) VOLUME: 261
(F) PAGES: 5341-5349
(G) DATE: 1986
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:22 Gly Phe Ala Ser Phe Leu Gly Lys Ala Leu
5 10
Lys Ala Ala Leu Lys lle Gly Ala Asn Ala
15 20
Leu Gly Gly Ala Pro Gln Gln
25
(2) INFORMATION FOR SEQ ID NO: 23:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 27 amino acids (B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(A) NAME/KEY: CPF peptide.
(x) PUBLICATION INFORMATION:
(A) AUTHOR: Richter, K.
Egger, R.
Kreil
(C) JOURNAL: J. Biol. Chem.
(D) VOLUME, 261
(F) PAGES: 3676-3680
( G ) DATE : 1986
(A) AUTHOR: Wakabayashi, T.
Kato, H.
Tachibaba, S.
(C) JOURNAL: Nucleic Acids Research
(D) VOLUME: 13
(F) PAGES: 1817-1828
(G) DATE: 1985
(A) AUTHOR: Gibson, B.W.
Poulter, L.
Williams, D.H.
Maggio, J.E.
(C) JOURNAL: J. Biol. Chem.
(D) VOLUME: 261
(F) PAGES: 5341-5349
(G) DATE: 1986 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:23: Gly Phe Ala Ser Phe Leu Gly Lys Ala Leu
5 10
Lys Ala Ala Leu Lys lle Gly Ala Asn Met
15 20
Leu Gly Gly Ala Pro Gln Gln
25
(2) INFORMATION FOR SEQ ID NO:24:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 27 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(A) NAME/KEY: CPF peptide.
(x) PUBLICATION INFORMATION:
(A) AUTHOR: Richter, K.
Egger, R.
Kreil
(C) JOURNAL: J. Biol. Chem..
(D) VOLUME: 261
(F) PAGES: 3676-3680
(G) DATE: 1986
(A) AUTHOR: Wakabayashi, T.
Kato, H.
Tachibaba, S.
(C) JOURNAL: Nucleic Acids Research
(D) VOLUME: 13
(F) PAGES: 1817-1828 ( G ) DATE : 1985
(A) AUTHOR: Gibson, B.W.
Poulter, L.
Williams, D.H.
Maggio, J.E.
(C) JOURNAL: J. Biol. Chem.
(D) VOLUME: 261
(F) PAGES: 5341-5349
(G) DATE: 1986
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:24: Gly Phe Gly Ser Phe Leu Gly Lys Ala Leu
5 10
Lys Ala Ala Leu Lys l le Gly Ala Asn Ala
15 20
Leu Gly Gly Ser Leu Gln Gln
25
(2) INFORMATION FOR SEQ ID NO: 25:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 27 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(A) NAME/KEY: CPF peptide.
(x) PUBLICATION INFORMATION:
(A) AUTHOR: Richter, K.
Egger, R.
Kreil (C) JOURNAL: J. Biol. Chem.
(D) VOLUME : 261
(F) PAGES: 3676-3680
(G) DATE: 1986
(A) AUTHOR: Wakabayashi, T.
Kato, H.
Tachibaba, S.
(C) JOURNAL: Nucleic Acids Research
(D) VOLUME: 13
(F) PAGES: 1817-1828
(G) DATE: 1985
(A) AUTHOR: Gibson, B.W.
Poulter, L.
Williams, D.H.
Maggio, J.E.
(C) JOURNAL: J. Biol. Chem.
(D) VOLUME: 261
(F) PAGES: 5341-5349
(G) DATE: 1986
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:25: Gly Phe Gly Ser Phe Leu Gly Lys Ala Leu
5 10
Lys Ala Gly Leu Lys lle Gly Thr Asn Phe
15 20
Leu Gly Gly Ala Pro Gln Gln
25
(2) INFORMATION FOR SEQ ID NO: 26:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 27 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS: (D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(A) NAME/KEY: CPF peptide.
(x) PUBLICATION INFORMATION:
(A) AUTHOR: Richter, K
Egger, R.
Kreil
(C) JOURNAL: J. Biol. Chem.
(D) VOLUME: 261
(F) PAGES: 3676-3680
(G) DATE: 1986
(A) AUTHOR: Wakabayashi, T.
Kato, H.
Tachibaba, S.
(C) JOURNAL: Nucleic Acids Research
(D) VOLUME: 13
(F) PAGES: 1817-1828
(G) DATE: 1985
(A) AUTHOR: Gibson, B.W.
Poulter, L.
Williams, D.H.
Maggio, J.E.
(C) JOURNAL: J. Biol. Chem.
(D) VOLUME: 261
(F) PAGES: 5341-5349
(G) DATE: 1986
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:26: Gly Leu Ala Ser Leu Leu Gly Lys Ala Leu
5 10 Lys Ala Ala Leu Lys lle Gly Ala Asn Ala
15 20
Leu Gly Gly Ser Pro Gln Gln
25
(2) INFORMATION FOR SEQ ID NO:27:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:27: Lys lle Ala Gly Lys lle Ala Lys lle Ala
5 10
Gly Lys lle Ala Lys lle Ala Gly Lys lle
15 20
Ala
(2) INFORMATION FOR SEQ ID NO: 28:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:28: Lys lle Ala Lys lle Ala Gly Lys lle Ala
5 10 Lys lle Ala Gly Lys lle Ala Lys lle Ala
15 20
Gly
(2) INFORMATION FOR SEQ ID NO: 29:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:29:
Lys lle Ala Gly Lys lle Gly Lys lle Ala
5 10
Gly Lys lle Gly Lys lle Ala Gly Lys lle
15 20
Gly
(2) INFORMATION FOR SEQ ID NO: 30:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:30: Lys Leu Ala Gly Lys Leu Ala Lys Leu Ala
5 10 Gly Lys Leu Ala Lys Leu Ala Gly Lys Leu
15 20
Ala
(2) INFORMATION FOR SEQ ID NO: 31:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:31: Lys Phe Ala Gly Lys Phe Ala Lys Phe Ala
5 10
Gly Lys Phe Ala Lys Phe Ala Gly Lys Phe
15 20
Ala
(2) INFORMATION FOR SEQ ID NO: 32:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:32: Lys Ala Leu Ser Lys Ala Leu Lys Ala Leu
5 10 Ser Lys Ala Leu Lys Ala Leu Ser Lys Ala
15 20
Leu
(2) INFORMATION FOR SEQ ID NO: 33:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:33: Lys Leu Leu Lys Ala Leu Gly Lys Leu Leu
5 10
Lys Ala Leu Gly Lys Leu Leu Lys Ala Leu
15 20
Gly
(2) INFORMATION FOR SEQ ID NO: 34:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:34: Lys Ala lle Gly Lys Ala lle Lys Ala lle
5 10 Gly Lys Ala lle Lys Ala lle Gly Lys Ala
15 20 lle
(2) INFORMATION FOR SEQ ID NO: 35:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:35: Gly lle Ala Lys lle Ala Lys Gly lle Ala
5 10
Lys lle Ala Lys Gly lle Ala Lys lle Ala
15 20
Lys
(2) INFORMATION FOR SEQ ID NO: 36:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:36: Lys lle Ala Lys lle Phe Gly Lys lle Ala
5 10 Lys lle Phe Gly Lys lle Ala Lys lle Phe
15 20
Gly
(2) INFORMATION FOR SEQ ID NO: 37:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:37: Gly lle Ala Arg lle Ala Lys Gly lle Ala
5 10
Arg lle Ala Lys Gly lle Ala Arg lle Ala
15 20
Lys
(2) INFORMATION FOR SEQ ID NO: 38:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:38: Lys Phe Ala Arg lle Ala Gly Lys Phe Ala
5 10 Arg lle Ala Gly Lys Phe Ala Arg lle Ala
15 20
Gly
(2) INFORMATION FOR SEQ ID NO: 39:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:39:
Gly Phe Ala Lys lle Ala Lys Gly Phe Ala
5 10
Lys lle Ala Lys Gly Phe Ala Lys lle Ala
15 20
Lys
(2) INFORMATION FOR SEQ ID NO: 40:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(D) OTHER INFORMATION: Xaa is ornithine
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:40:
Lys lle Ala Gly Xaa lle Ala Lys lle Ala
5 10 Gly Xaa lle Ala Lys lle Ala Gly Xaa lle
15 20
Ala
(2) INFORMATION FOR SEQ ID NO: 41:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:41:
Lys lle Ala Arg lle Ala Gly Lys lle Ala
5 10
Arg lle Ala Gly Lys lle Ala Arg lle Ala
15 20
Gly
(2) INFORMATION FOR SEQ ID NO: 42:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(D) OTHER INFORMATION: Xaa is ornithine (xi) SEQUENCE DESCRIPTION: SEQ ID NO:42:
Xaa lle Ala Gly Lys lle Ala Xaa lle Ala
5 10
Gly Lys lle Ala Xaa lle Ala Gly Lys lle
15 20
Ala
(2) INFORMATION FOR SEQ ID NO: 43:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:43:
Gly lle Ala Arg lle Phe Lys Gly lle Ala
5 10
Arg lle Phe Lys Gly lle Ala Arg lle Phe
15 20
Lys
(2) INFORMATION FOR SEQ TD NO: 44:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(D) OTHER INFORMATION: Xaa is norleucine, (xi) SEQUENCE DESCRIPTION: SEQ ID NO:44:
Lys Xaa Ala Gly Lys Xaa Ala Lys Xaa Ala
5 10
Gly Lys Xaa Ala Lys Xaa Ala Gly Lys Xaa
15 20
Ala
(2) INFORMATION FOR SEQ ID NO: 45:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(D) OTHER INFORMATION: Xaa is norleucine.
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:45:
Lys Xaa Ala Gly Lys lle Ala Lys Xaa Ala
5 10
Gly Lys lle Ala Lys Xaa Ala Gly Lys lle
15 20
Ala
(2) INFORMATION FOR SEQ ID NO:46:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide (ix) FEATURE:
(D) OTHER INFORMATION: Xaa is norleucine,
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:46:
Lys lle Ala Gly Lys Xaa Ala Lys lle Ala
5 10
Gly Lys Xaa Ala Lys lle Ala Gly Lys Xaa
15 20
Ala
(2) INFORMATION FOR SEQ ID NO:47:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(D) OTHER INFORMATION: Xaa is norvaline.
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:47:
Lys Xaa Ala Gly Lys Xaa Ala Lys Xaa Ala
5 10
Gly Lys Xaa Ala Lys Xaa Ala Gly Lys Xaa
15 20
Ala
(2) INFORMATION FOR SEQ ID NO:48:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS: (D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(D) OTHER INFORMATION: Xaa is norvaline.
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:48:
Lys Xaa Ala Gly Lys lle Ala Lys Xaa Ala
5 10
Gly Lys lle Ala Lys Xaa Ala Gly Lys Xaa
15 20
Ala
(2) INFORMATION FOR SEQ ID NO:49:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:49:
Lys Leu Leu Ser Lys Leu Gly Lys Leu Leu
5 10
Ser Lys Leu Gly Lys Leu Leu Ser Lys Leu
15 20
Gly
(2) INFORMATION FOR SEQ ID NO:50:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid (C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:50:
Lys Leu Leu Ser Lys Phe Gly Lys Leu Leu
5 10
Ser Lys Phe Gly Lys Leu Leu Ser Lys Phe
15 20
Gly
(2) INFORMATION FOR SEQ ID NO:51:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(D) OTHER INFORMATION: Xaa is norvaline,
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:51:
Lys lle Ala Gly Lys Xaa Ala Lys lle Ala
5 10
Gly Lys Xaa Ala Lys lle Ala Gly Lys Xaa
15 20
Ala
(2) INFORMATION FOR SEQ ID NO: 52:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids (B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:52: His lle Ala Gly His lle Ala His lle Ala
5 10
Gly His lle Ala His lle Ala Gly His lle
15 20
Ala
(2) INFORMATION FOR SEQ ID NO:53:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:53:
Ala Gly Lys lle Ala Lys lle Ala Gly Lys
5 10
lle Ala Lys lle Ala Gly Lys lle Ala Lys
15 20 lle
(2) INFORMATION FOR SEQ ID NO: 54:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS: (D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:54: lle Ala Lys lle Ala Gly Lys lle Ala Lys
5 10
lle Ala Gly Lys lle Ala Lys lle Ala Gly
15 20
Lys
(2) INFORMATION FOR SEQ ID NO: 55:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:55: Lys lle Ala Gly Arg lle Ala Lys lle Ala
5 10
Gly Arg lle Ala Lys lle Ala Gly Arg lle
15 20
Ala
(2) INFORMATION FOR SEQ ID NO: 56:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:56: Arg lle Ala Gly Arg lle Ala Arg lle Ala
5 10
Gly Arg lle Ala Arg lle Ala Gly Arg lle
15 20
Ala
(2) INFORMATION FOR SEQ ID NO: 57:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide"
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:57: Lys Val Ala Gly Lys lle Ala Lys Val Ala
5 10
Gly Lys lle Ala Lys Val Ala Gly Lys lle
15 20
Ala
(2) INFORMATION FOR SEQ ID NO: 58:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:58:
Lys lle Ala Gly Lys Val Ala Lys lle Ala
5 10
Gly Lys Val Ala Lys lle Ala Gly Lys Val
15 20
Ala
(2) INFORMATION FOR SEQ ID NO: 59:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:59:
Ala Lys lle Ala Gly Lys lle Ala Lys lle
5 10
Ala Gly Lys lle Ala Lys lle Ala Gly Lys
15 20
lle
(2) INFORMATION FOR SEQ ID NO: 60:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(D) OTHER INFORMATION: Xaa is ornithine. (xi) SEQUENCE DESCRIPTION: SEQ ID NO:60: Xaa lle Ala Gly Xaa lle Ala Xaa lle Ala
5 10
Gly Xaa lle Ala Xaa lle Ala Gly Xaa lle
15 20
Ala
(2) INFORMATION FOR SEQ ID NO: 61:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:61: Lys Phe Ala Gly Lys lle Ala Lys Phe Ala
5 10
Gly Lys lle Ala Lys Phe Ala Gly Lys lle
15 20
Ala
(2) INFORMATION FOR SEQ ID NO:62:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH : 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:62 :
Lys lle Ala Gly Lys Phe Ala Lys lle Ala
5 10
Gly Lys Phe Ala Lys lle Ala Gly Lys Phe
15 20
Ala
(2) INFORMATION FOR SEQ ID NO: 63:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(D) OTHER INFORMATION: Xaa is cyclohexylalanine
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:63:
Lys Xaa Ala Gly Lys lle Ala Lys Xaa Ala
5 10
Gly Lys lle Ala Lys Xaa Ala Gly Lys lle
15 20
Ala
(2) INFORMATION FOR SEQ ID NO: 64:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide ( ix ) FEATURE :
(D) OTHER INFORMATION: Xaa is norleucine.
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:64:
Lys Xaa Ala Lys lle Ala Gly Lys Xaa Ala
5 10
Lys lle Ala Gly Lys Xaa Ala Lys lle Ala
15 20
Gly
(2) INFORMATION FOR SEQ ID NO:65:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 AMINO ACIDS
(B) TYPE: amino acids
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:65:
Arg lle Ala Gly Lys lle Ala Arg lle Ala
5 10
Gly Lys lle Ala Arg lle Ala Gly Lys lle
15 20
Ala
(2) INFORMATION FOR SEQ ID NO: 66:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOFOLOGY: linear
(ii) MOLECULE TYPE: peptide ( ix ) FEATURE :
(D) OTHER INFORMATION: Xaa is homoarginine.
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:66:
Xaa lle Ala Gly Xaa lle Ala Xaa lle Ala
5 10
Gly Xaa lle Ala Xaa lle Ala Gly Xaa lle
15 20
Ala
(2) INFORMATION FOR SEQ ID NO: 67:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE: Xaa is p-aminophenylalanine
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:67:
Xaa lle Ala Gly Lys lle Ala Xaa lle Ala
5 10
Gly Lys lle Ala Xaa lle Ala Gly Lys lle
15 20
Ala
(2) INFORMATION FOR SEQ ID NO: 68:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide
(ix) FEATURE: Xaa is p-aminophenylalanine
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:68: Lys lle Ala Gly Xaa lle Ala Lys lle Ala
5 10
Gly Xaa lle Ala Lys lle Ala Gly Xaa lle
15 20
Ala
(2) INFORMATION FOR SEQ ID NO:69:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 24 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:69: Lys lle Ala Lys lle Ala Gly Lys lle Ala
5 10
Lys lle Ala Gly Lys lle Ala Lys lle Ala
15 20
Gly Lys lle Ala
(2) INFORMATION FOR SEQ ID NO: 70:
(i) SEQUENCE CHARACTERI STICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:70:
Lys Leu Ala Ser Lys Ala Gly Lys lle Ala Gly
5 10
Lys lle Ala Lys Val Ala Leu Lys Ala Leu
15 20
(2) INFORMATION FOR SEQ ID NO:71:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(D) OTHER INFORMATION: Xaa is ornithine,
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 71:
Lys lle Ala Gly Lys lle Ala Lys lle Ala Gly
5 10
Xaa lle Ala Lys lle Ala Gly Lys lle Ala
15 20
(2) INFORMATION FOR SEQ ID NO:72:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:72:
Lys lle Ala Gly Lys lle Ala Lys lle Ala
5 10
Gly Arg lle Ala Lys lle Ala Gly Lys lle
15 20
Ala
(2) INFORMATION FOR SEQ ID NO:73:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(D) OTHER INFORMATION: Xaa is norleucine
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:73:
Lys lle Ala Gly Lys lle Ala Lys lle Ala
5 10
Gly Xaa lle Ala Lys lle Ala Gly Lys lle
15 20
Ala
(2) INFORMATION FOR SEQ ID NO:74:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide ( ix ) FEATURE :
(D) OTHER INFORMATION: Xaa is norvaline.
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:74:
Lys lle Ala Gly Lys lle Ala Lys lle Ala
5 10
Gly Xaa lle Ala Lys lle Ala Gly Lys lle
15 20
Ala
(2) INFORMATION FOR SEQ ID NO:75:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(D) OTHER INFORMATION: Xaa is ornithine.
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:75:
Lys Phe Ala Gly Lys Phe Ala Lys Phe Ala Gly
5 10
Xaa Phe Ala Lys Phe Ala Gly Lys Phe Ala
15 20
(2) INFORMATION FOR SEQ ID NO: 76:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(D) OTHER INFORMATION: Xaa is ornithine.
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:76:
Lys lle Ala Gly Lys Phe Ala Lys lle Ala
5 10
Gly Xaa Phe Ala Lys lle Ala Gly Lys Phe
15 20
Ala
(2) INFORMATION FOR SEQ ID NO:77:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(D) OTHER INFORMATION: Xaa at residues 6, 13, and 20 is norleucine; Xaa at residue
12 is ornithine.
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 77:
Lys lle Ala Gly Lys Xaa Ala Lys lle Ala
5 10
Gly Xaa Xaa Ala Lys lle Ala Gly Lys Xaa
15 20
Ala
(2) INFORMATION FOR SEQ ID NO:78: (i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:78: Lys Met Ala Ser Lys Ala Gly Lys lle Ala
5 10
Gly Lys lle Ala Lys Val Ala Leu Lys Ala
15 20
Leu
(2) INFORMATION FOR SEQ ID NO: 79:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:79 Lys lle Ala Ser Lys Ala Gly Lys lle Ala
5 10
Gly Lys lle Ala Lys Val Ala Leu Lys Ala Leu
15 20
(2) INFORMATION FOR SEQ ID NO:80:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid (C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(D) OTHER INFORMATION: Xaa is norleucine.
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:80:
Lys lle Ala Ser Lys Ala Gly Lys Xaa Ala
5 10
Gly Lys lle Ala Lys Val Ala Leu Lys Ala Leu
15 20
(2) INFORMATION FOR SEQ ID NO:81:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(D) OTHER INFORMATION: Xaa is norleucine.
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:81:
Lys Leu Ala Ser Lys Ala Gly Lys Xaa Ala
5 10
Gly Lys lle Ala Lys Val Ala Leu Lys Ala
15 20
Leu
(2) INFORMATION FOR SEQ ID NO:82: (i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(D) OTHER INFORMATION: Xaa is norleucine.
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:82:
Lys Xaa Ala Ser Lys Ala Gly Lys Xaa Ala
5 10
Gly Lys lle Ala Lys Val Ala Leu Lys Ala Leu
15 20
(2) INFORMATION FOR SEQ ID NO:83:
(i) SEQUENCE CHARACTERISTICS :
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(D) OTHER INFORMATION: Xaa is p-aminophenylal anine .
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:83: Lys lle Ala Gly Lys lle Ala Lys lle Ala
5 10
Gly Xaa lle Ala Lys lle Ala Gly Lys lle
15 20
Ala
(2) INFORMATION FOR SEQ ID NO:84:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:84: Lys lle Ala Gly Ala lle Ala Lys lle Ala
5 10
Gly Lys lle Ala Lys lle Ala Gly Lys lle
15 20
Ala
(2) INFORMATION FOR SEQ ID NO:85:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:85: Lys lle Ala Gly Lys lle Ala Lys lle Ala
5 10
Gly Ala lle Ala Lys lle Ala Gly Lys Ile
15 20
Ala
(2) INFORMATION FOR SEQ ID NO:86:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:86: Lys lle Ala Gly Lys lle Ala Lys lle Ala
5 10
Gly Lys lle Ala Lys lle Ala Gly Ala lle
15 20
Ala
(2) INFORMATION FOR SEQ ID NO:87:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:87: Lys Leu Ala Ser Lys Ala Ala Lys lle Ala 5 10
Ala Lys lle Ala Lys Val Ala Leu Lys Ala
10 20
Leu
(2) INFORMATION FOR SEQ ID NO:88:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:88: Lys lle Ala Lys Lys lle Ala Lys lle Ala
5 10
Lys Lys lle Ala Lys lle Ala Lys Lys lle
15 20
Ala
(2) INFORMATION FOR SEQ ID NO: 89:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:89: Lys Phe Ala Lys Lys Phe Ala Lys Phe Ala
5 10
Lys Lys Phe Ala Lys Phe Ala Lys Lys Phe 15 20
Al a
(2) INFORMATION FOR SEQ ID NO:90:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:90: Lys Phe Ala Lys Lys lle Ala Lys Phe Ala
5 10
Lys Lys lle Ala Lys Phe Ala Lys Lys lle
15 20
Ala
(2) INFORMATION FOR SEQ ID NO:91:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:91: Ala lle Ala Gly Lys lle Ala Lys lle Ala
5 10
Gly Lys lle Ala Lys lle Ala Gly Lys lle
15 20
Ala (2) INFORMATION FOR SEQ ID NO:92:
(j) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:92: Lys lle Ala Gly Lys lle Ala Ala lle Ala
5 10
Gly Lys lle Ala Lys lle Ala Gly Lys lle
15 20
Ala
(2) INFORMATION FOR SEQ ID NO:93:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:93: Lys lle Ala Gly Lys lle Ala Lys lle Ala
5 10
Gly Lys lle Ala Ala lle Ala Gly Lys lle
15 20
Ala
(2) INFORMATION FOR SEQ ID NO:94:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 21 amino acids
(B) TYPE: .amino acid (C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:94: Gly Met Ala Ser Lys Ala Gly Lys lle Ala
5 10
Gly Lys lle Ala Lys Val Ala Leu Lys Ala
15 20
Leu
(2) INFORMATION FOR SEQ ID NO:95:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 11 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:95: Leu Lys Lys Leu Lys Lys Leu Leu Lys Leu
5 10
Leu
(2) INFORMATION FOR SEQ ID NO:96:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 12 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:96: Leu Leu Lys Lys Leu Lys Lys Leu Leu Lys
5 10
Leu Leu
(2) INFORMATION FOR SEQ ID NO:97:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 13 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:97:
Lys Leu Leu Lys Lys Leu Lys Lys Leu Leu
5 10
Lys Leu Leu
(2) INFORMATION FOR SEQ ID NO:98:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 14 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:98:
Lys Lys Leu Leu Lys Lys Leu Lys Lys Leu
5 10
Leu Lys Leu Leu (2) INFORMATION FOR SEQ ID NO:99:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 16 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) SEQUENCE DESCRIPTION: SEQ ID NO:99:
Lys Lys Leu Leu Lys Lys Leu Lys Lys Leu Leu Lys Lys Leu Arg Arg
5 10 15
(2) INFORMATION FOR SEQ ID NO:100:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 16 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:100:
Lys Leu Lys Lys Leu Leu Lys Lys Leu Lys
5 10
Lys Leu Leu Lys Leu Leu
15
(2) INFORMATION FOR SEQ ID NO:101:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 15 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS: (D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:101: Leu Lys Lys Leu Leu Lys Lys Leu Lys Lys
5 10
Leu Leu Lys Lys Asn
15
(2) INFORMATION FOR SEQ ID NO:102:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 15 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(D) OTHER INFORMATION: Xaa is homoserine
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:102: Leu Lys Lys Leu Leu Lys Lys Leu Lys Lys
5 10
Leu Leu Lys Lys Xaa
15
(2) INFORMATION FOR SEQ ID NO:103:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 18 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:103 : Leu Lys Leu Leu Lys Lys Leu Leu Lys Lys
5 10
Asn Lys Lys Leu Leu Lys Lys Leu
15
(2) INFORMATION FOR SEQ ID NO:104:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 18 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEO ID NO:104: Leu Lys Leu Leu Lys Lys Leu Leu Lys Lys
5 10
Pro Lys Lys Leu Leu Lys Lys Leu
15
(2) INFORMATION FOR SEQ ID NO:105:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 22 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:105: Leu Leu Lys Lys Leu Lys Lys Leu Leu Lys
5 10
Lys Leu Gln Gly Pro Pro Gln Gly Gln Ser
15 20
Pro Gln
(2) INFORMATION FOR SEQ ID NO:106:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 20 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:106: Leu Ala Ser Lys Ala Gly Ala lle Ala Gly
5 10
Lys lle Ala Lys Lys Leu Leu Lys Lys Leu
15 20
(2) INFORMATION FOR SEQ ID NO:107:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 7 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:107: Leu Lys Lys Leu Lys Lys Leu
5 (2) INFORMATION FOR SEQ ID NO:108:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 8 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:108: Leu Leu Lys Lys Leu Lys Lys Leu
5
(2) INFORMATION FOR SEQ ID NO: 109:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 9 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:109: Lys Leu Leu Lys Lys Leu Lys Lys Leu
5
(2) INFORMATION TOR SEQ ID NO:110:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 10 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:110: Lys Lys Leu Leu Lys Lys Leu Lys Lys Leu
5 10
(2) INFORMATION FOR SEQ ID NO: 111:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 11 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:111: Leu Lys Lys Leu Leu Lys Lys Leu Lys Lys Leu
5 10
(2) INFORMATION FOR SEQ ID NO: 112:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 11 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:112: Ala Lys Lys Leu Leu Lys Lys Leu Lys Lys Leu
5 10
(2) INFORMATION FOR SEQ ID NO: 113:
(i) SEQUENCE CHARACTERISTICS
(A) LENGTH: 14 amino acids
(B) TYPE: amino acid (C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:113:
Leu Lys Lys Leu Leu Lys Lys Leu Lys Lys Leu
5 10
Leu Lys Arg
(2) INFORMATION FOR SEQ ID NO:114:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 26 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(vi) ORIGINAL SOURCE
(A) ORGANISM: Apis mellifera
(vii) FEATURE
(A) NAME/KEY: melittin peptide
(x) PUBLICATION INFORMATION:
(A) AUTHORS: Habermann, E.
Jentsch, J.
(B) TITLE: Sequenzanalyse des Melittins aus den tryptischen and peptischen
Spaltstucken
(C) JOURNAL: Hoppe-Seyler's Zeitschrift
Physiol. Chem. (D) VOLUME: 348
(F) PAGES: 37-50
(G) DATE: 1987
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:114:
Gly lle Gly Ala Val Leu Lys Val Leu
5
Thr Thr Gly Leu Pro Ala Leu lle Ser Trp
10 15
lle Lys Arg Lys Arg Gln Gln
20 25
(2) INFORMATION FOR SEQ ID NO:115:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 18 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:115: Leu Lys Leu Leu Lys Lys Leu Leu Lys Lys
5 10
Leu Lys Lys Leu Leu Lys Lys Leu
15
(2) INFORMATION FOR SEQ ID NO:116:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 15 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(D) OTHER INFORMATION: C-terminal amide, may be acetylated at N-terminus.
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:116:
Leu Lys Lys Leu Leu Lys Lys Leu Lys Lys
5 10
Leu Leu Lys Lys Leu
15
(2) INFORMATION FOR SEQ ID NO:117:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 14 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS :
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(D) OTHER INFORMATION: C-terminal amide, may be arptylated at N-terminus.
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:117:
Lys Lys Leu Leu Lys Lys Leu Lys Lys Leu
5 10
Leu Lys Lys Leu
(2) INFORMATION FOR SEQ ID NO:118:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 14 amino acids
(B) TYPE: amino acid (C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:118: Leu Leu Lys Leu Leu Lys Lys Leu Leu Lys
5 10
Lys Leu Lys Lys
(2) INFORMATION FOR SEQ ID NO: 119:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 14 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEO ID NO:119: Lys Leu Leu Lys Lys Leu Lys Lys Leu Leu
5 10
Lys Lys Leu Leu
(2) INFORMATION FOR SEQ ID NO:120:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 16 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS:
(D) TOPOLOGY: linear
(ii) MOLECULE TYFE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:120: Leu Leu Lys Lys Leu Lys Lys Leu Leu Lys
5 10
Lys Leu Leu Lys Leu Leu
15

Claims

WHAT IS CLAIMED IS:
1. A process for treating a cancerous growth in a host, comprising:
administering to a host (a) at least one biologically active amphiphilic ion channel-forming peptide or protein; and (b) at least one protease inhibitor, said at least one peptide or protein and said at least one protease inhibitor being administered in amounts
effective to treat a cancerous growth in a host.
2. The process of Claim 1 wherein said peptide or protein is selected from the group consisting of:
(a) magainin peptides;
(b) PGLa peptides;
(c) XPF peptides;
(d) CPF peptides;
(e) cecropins;
(f) sarcotoxins;
(g) a peptide including one of the following basic structures X31 through X37 wherein:
X31 is -[R31-R32-R32-R33-R31-R32-R32]n-;
X32 is -(R32-R32-R 33-R31-R32-R32-R31]n -;
X33 is -[R32-R33-R31-R32-R32-R31-R32]n-;
X34 is -[R33-R31-R32-R32-R31-R32-R32]n-;
X35 is -[R31-R32-R32-R31-R32-R32-R33]n-;
and
X36 is -[R32-R32-R31-R32-R32-R33-R31]n-;
X37 is -[R32-R31-R32-R32-R33-R31-R32]n-;
wherein R31 is a basic hydrophilic amino acid, R32 is a
hydrophobic amino acid, R33 is a neutral hydrophilic, basic
hydrophilic. or hydrophobic amino acid, and n is from 2 to 5;
(h) a peptide including the following basic structure X40: R31-R32-R32-R33-R33-R32-R32-R31-R32-R32-R32-R34-R32-R32,whereir R31 is a basic hydrophilic amino acid, R32 is a hydrophobic amin acid, R33 is a neutral hydrophilic or hydrophobic amino acid, an is a basic hydrophilic or hydrophobic amino acid;
(i) a peptide including the following basic structure X50: R41-R42-R42-R41-R42-R42-R41-R41-R42-R41-R41-, wherein R41 is a hydrophobic amino acid, and R42 is a basic hydrophilic or neutral hydrophobic amino acid;
(j) a peptide including the following basic structure X52 :
R42-R41-R42-R41-R42-R41-R42-R42-R41-R42-R42-, wherein R41 is a hydrophobic amino acid, and R42 is a basic hydrophilic or neutral hydrophilic amino acid;
(k) a peptide including the following basic structure X54:
R41-R42-R42-R41-R41-R42-R42-R41-R42-R42-R41-R41-
R42-R42-R43-, wherein R41 is a hydrophobic amino acid, R42 is a basic hydrophilic or neutral hydrophilic amino acid, and R43 is a neutral hydrophilic amino acid;
(l) a peptide including the following basic structure X56 :
-R41-R42-R41-R41-R42-R42-R41-R41-R42-R42-R44-, wherein R41 is a hydrophobic amino acid, R42 is a basic hydrophilic or neutral
hydrophilic amino acid, and R44 is a neutral hydrophilic amino acid o proline;
(m) a peptide including the following basic structure X58:
-R41-R41-R42-R42-R41-R42-R42-R41-R41-R42-R42-R41-R43, where is a hydrophobic amino acid, R42 is a basic hydrophilic or neutral hydrophilic amino acid, and R43 is a neutral hydrophilic amino acid;
(n) a peptide including the following basic structure X60:
-R41-R41-R43-R42-R41-R41-R41-R41-R41-R41-R42-R41-R41- R42-R42-R41-R41-R42-R42-R42-R41, wherein R41 is a hydrophobic amino acid, R42 is a basic hydrophilic or neutral hydrophilic amino acid, and R43 is a neutral hydrophilic amino acid;
(o) a peptide having a structure selected from the group consisting of:
(i) R41-R42-R42-R41-R42-R42-R41;
(ii) R41-R41-R42-R42-R41-R42-R42-R41;
(iii) R42-R41-R41-R42-R42-R41-R42-R42-R41;
(iv) R42-R42-R41-R41-R42-R42-R41-R42-R42-R41; and (v) R41-R42-R42-R41-R41-R42-R42-R41-R42-R42-R41, wherein R41 i a hydrophobic amino acid, and R42 is a basic hydrophilic amino acid c a neutral hydrophilic amino acid;
(p) a peptide, said peptide being an amide - or
carboxy-terminated peptide represented by the following structural formula, and the numbers below each amino acid residue refer to the position of the residue in the peptide:
Leu Lys Leu Leu Lys Lys Leu Leu Lys Lys Leu
1 2 3 4 5 6 7 8 9 10 11
Lys Lys Leu Leu Lys Lys Leu
12 13 14 15 16 17 18
or an analogue of said peptide wherein at least one of amino acid residues 1 through 7, 9, 11, 12, 14, 16, or 18 is deleted from the peptide;
(q) a peptide including the following structural formula X62:
R41-R41-R42-R41-R41-R42-R42-R41-R41-R42-R42-R41-R42-R42, wherein R41 is a hydrophobic amino acid, and R42 is a basic
hydrophilic or neutral hydrophilic amino acid;
(r) a peptide including the following structural formula X64:
R42-R41-R41-R42-R42-R41-R42-R42-R41-R41-R42-R42-R41-R41, wherein R41 is a hydrophobic amino acid, and R42 is a basic
hydrophilic or neutral hydrophilic amino acid;
(s) a peptide including the following structural formula X66:
R41-R41-R42-R42-R41-R42-R42-R41-R41-R42-R42-R41-R41,
wherein R41 is a hydrophobic amino acid, and R42 is a basic
hydrophilic or neutral hydrophilic amino acid;
(t) melittin;
(u) apidaecins;
(v) defensins;
(w) major basic protein of eosinophils;
(x) bacterial permeability-increasing protein; and
(y) perforin.
3. The process of Claim 2 wherein the peptide is a magainin peptide.
4. The process of Claim 2 wherein the peptide is a PGLa peptide,
5. The process of Claim 2 wherein the peptide is an XPF peptide.
6. The process of Claim 2 wherein the peptide is a CPF peptide
7. The process of Claim 2 wherein the peptide is a cecropin .
8. The process of Claim 2 wherein the peptide is a sarcotoxin.
9. The process of Claim 2 wherein the peptide includes one of the following basic structures X31 through X37, wherein:
X31 is -[R31-R32-R32-R33-R31-R32-R32]n-;
X32 is -[R32-R32-R33-R31-R32-R32-R31]n-;
X33 is -[R32-R33-R31-R32-R32-R31-R32]n-;
X34 is -[R33-R31-R32-R32-R31-R32-R32]n-;
X35 is -[R31-R32-R32-R31-R32-R32-R33]n-;
X36 is -[R32-R32-R31-R32-R32-R33-R31]n-; and
X37 is -[R32-R31-R32-R32-R33-R31-R32]n-; wherein R31 is a basic hydrophilic amino acid, R32 is a hydrophobic amino acid, R33 is a neutral hydrophilic, basic hydrophilic, or hydrophobic amino acid, and n is from 2 to 5.
10. The process of Claim 2 wherein the peptide includes the following basic structure X40:
R31-R32-R32-R33-R34-R32-R32-R31-R32-R32-R32-R34-R32-R32,
wherein R31 is a basic hydrophilic amino acid, R32 is a hydrophobic amino acid, R33 is a neutral hydrophilic or hydrophobic amino acid, and R34 is a basic hydrophilic or hydrophobic amino acid.
11. The process of Claim 2 wherein said peptide includes the following basic structure X50:
R41-R42-R42-R41-R42-R42-R41-R41-R42-R41-R41,
wherein R41 is a hydrophobic amino acid, and R42 is a basic
hydrophilic or neutral hydrophilic ammo acid.
12. The process of Claim 2 wherein said peptide includes the following basic structure X52:
R42-R41-R42-R42-R41-R41-R42-R42-R41-R42-R42, wherein R41 is a hydrophobic amino acid, and R42 is a basic
hydrophilic or neutral hydrophilic amino acid.
13. The process of Claim 2 wherein the peptide includes the following basic structure X54:
-R41-R42-R42-R41-R41-R42-R42-R41-R42-R42-R41-R41-R42-R42-R43-, wherei R41 is a hydrophobic amino acid, R42 is a basic hydrophilic or neutra hydrophilic amino acid, and R43 is a neutral hydrophilic amino acid.
14. The process of Claim 2 wherein the peptide includes the following basic structure X56:
-R41-R42-R41-R41-R42-R42-R41-R41-R42-R41-R4-, wherein R41 is a hydrophobic amino acid, R42 is a basic hydrophilic or neutral
hydrophilic amino acid, and R44 is a neutral hydrophilic amino acid o proline.
15. The process of Claim 2 wherein the peptide includes the following basic structure X58:
-R41-R41-R42-R42-R41-R42-R42-R41-R41-R42-R42- R41-R43-, wherein R41is a hydrophobic amino acid, R42 is a basic hydrophilic or neutral hydrophilic amino acid, and R43 is a neutral hydrophilic amino acid.
16. The process of Claim 2 wherein the peptide includes the following basic structure X60:
-R41-R41-R43-R42-R41-R41-R41-R41-R41-R41-R42-R41-R41-R42-R42R41- R41-R42-R42-R42-R41-, wherein R41 is a hydrophobic amino acid, R42is a basic hydrophilic or neutral hydrophilic amino acid, and R43 is a neutral hydrophilic amino acid.
17. The process of Claim 2 wherein the peptide has a structure selected from the group consisting of:
(i) R41-R42-R42-R41-R42-R42-R41;
(ii) R41-R41-R42-R42-R41-R42-R42-R41;
(iii) R42-R41-R41-R42-R42-R41-R42-R42-R41;
(iv) R42-R42-R41-R41-R42-R42-R41-R42-R42-R41; and
(v) R41-R42-R42-R41-R41-R42-R42-R41-R42-R42-R41, wherein R41 is a hydrophobic amino acid, and R42 is a basic hydrophilic aminc acid or a neutral hydrophilic amino acid.
18. The process of Claim 2 wherein said peptide is an amide-c carboxy-terminated peptide represented by the following structural formula, and the numbers below each amino acid residue refer to the position of the residue in the peptide:
Leu Lys Leu Leu Lys Lys Leu Leu Lys Lys Leu
1 2 3 4 5 6 7 8 9 10 11
Lys Lys Leu Leu Lys Lys Leu
12 13 14 15 16 17 18
or an analogue of said peptide wherein at least one of amino acid residues 1 through 7, 9, 11, 12, 14, 16, or 18 is deleted from the peptide.
19. The process of Claim 2 wherein said peptide includes the following structural formula X62:
-R41-R41-R42-R41-R41-R42-R42-R41-R41-R42-R42-R41-R42-R42-R42-, wherein R41 is a hydrophobic amino acid, and R42 is a basic
hydrophilic or neutral hydrophilic amino acid.
20. The process of Claim 2 wherein said peptide includes the following structural formula X64:
-R42-R41-R41-R42-R42-R41-R42-R42-R41-R41-R42-R42-R41-R41, wherein R41 is a hydrophobic amino acid, and R42 is a basic
hydrophilic or neutral hydrophilic amino acid.
21. The process of Claim 2 wherein said- peptide includes the following structural formula X66:
-R41-R42-R42-R41-R41-R42-R42-R41-R42-R42-R42-R41-R41,
wherein R41 is a hydrophobic amino acid, and R42 is a basic
hydrophilic or neutral hydrophilic amino acid.
22. The process of Claim 2 wherein said peptide is melittin.
23. The process of Claim 2 wherein said peptide is an apidaeci
24. The process of Claim 2 wherein said peptide or protein is defensin.
25. The process of Claim 2 wherein said peptide or protein is major basic protein of eosinophils.
26. The process of Claim 2 wherein said peptide or protein is bacterial permeability-increasing protein.
27. The process of Claim 2 wherein said peptide or protein is perforin.
28. The process of Claim 2 wherein the peptide is administered intralesionally.
29. The process of Claim 1 wherein said at least one protease inhibitor is bestatin.
30. The process of Claim 1 wherein each of said components (a) and (b) is administered in an amount ineffective to treat a cancerous growth in a host if administered alone to a host.
31. A composition for treating a cancerous growth in a host, comprising:
(a) at least one biologically active amphiphilic peptide c protein, said at least one peptide or protein being an ion
channel-forming peptide or protein;
(b) at least one protease inhibitor; and
(c) a physiologically acceptable pharmaceutical carrier, wherein components (a) and (b) are present in amounts effective to treat a cancerous growth in a host.
32. The composition of Claim 31 wherein said peptide or proteir is selected from the group consisting of:
(a) magainin peptides;
(b) PGLa peptides;
(c) XPF peptides;
(d) CPF peptides;
(e) cecropins;
(f) sarcotoxins;
(g) a peptide including one of the following basic structures
X31 through X37, wherein:
X31 is -[R31-R32-R32-R33-R31-R32-R32]n-;
X32 is -[R32-R32-R33-R31-R32-R32-R31]n-;
X33 is -[R32-R33-R31-R32-R32-R31-R32]n-;
X34 is -[R33-R31-R32-R32-R31-R32-R32]n-; X35 is -[R31-R32-R32-R31-R32-R32-R33]n-;
X36 is -[R32-R32-R31-R32-R32-R33-R31]n-; and
X37 is -[R32-R31-R32-R32-R33-R31-R32]n-;
wherein R31 is a basic hydrophilic amino acid, R32 is a
hydrophobic amino acid, R33 is a neutral hydrophilic, basic
hydrophilic, or hydrophobic amino acid, and n is from 2 to 5;
(h) a peptide including the following basic structure X40 :
R31-R32-R32-R33-R33-R32-R32-R31-R32-R32-R32-R34-R32-R32, wherei: R31 is a basic hydrophilic amino acid, R32 is a hydrophobic amino acid, R33 is a neutral hydrophilic or hydrophobic amino acid, and R3 is a basic hydrophilic or hydrophobic amino acid;
(i) a peptide including the following basic structure X50:
-R41-R42-R42-R41-R42-R42-R41-R41-R42-R41-R41-, Wherein R41 is a hydrophobic amino acid, and R42 is a basic hydrophilic or neutral hydrophobic amino acid;
(j) a peptide including the following basic structure X52:
-R42-R41-R42-R42-R41-R42-R42-R41-R42-R42-R42-, wherein R41 is a hydrophobic amino acid, and R42 is a basic hydrophilic or neutral hydrophilic amino acid;
(k) a peptide including the following basic structure X54:
-R41-R42-R42-R41-R41-R42-R42-R41-R42-R42-R41-R41- R 42-R42-R43-, wherein R41 is a hydrophobic amino acid, R42 is a basi hydrophilic or neutral hydrophilic amino acid, and R43 is a neutral hydrophilic amino acid;
(1) a peptide including the following basic structure X56:
-R41-R42-R41-R41-R42-R42-R41-R41-R42-R42-R44-, wherein R41 is a hydrophobic amino acid, R42 is a basic hydrophilic or neutral hydrophilic amino acid, and R44 is a neutral hydrophilic amino acid proline;
(m) a peptide including the following basic structure X58 : -R41-R41-R42-R42-R41-R42-R42-R41-R41-R42-R42-R41-R43, wherein R is a hydrophobic amino acid, R42 is a basic hydrophilic or neutral hydrophilic amino acid, and R43 is a neutral hydrophilic amino acid;
(n) a peptide including the following basic structure X60: -R41-R41-R43-R42-R41-R41-R41-R41-R41-R41-R42-R41-R41- R42-R42-R41-R41-R42-R42-R42-R41, wherein R41 is a hydrophobic amino acid, R42 is a basic hydrophilic or neutral hydrophilic amino acid, and R43 is a neutral hydrophilic amino acid;
(o) a peptide having a structure selected from the group consisting of:
(i) R41-R42-R42-R41-R42-R42-R41;
(ii) R41-R41-R42-R42-R41-R42-R42-R41;
(iii ) R42-R41-R41-R42-R42-R41-R42-R42-R41;
(iv) R42-R42-R41-R41-R42-R42-R41-R42-R42-R41; and
(v) R41-R42-R42-R41-R41-R42-R42-R41-R42-R42-R41, whereinR41i a hydrophobic amino acid, and R42 is a basic hydrophilic amino o a neutral hydrophilic amino acid;
(p) a peptide, said peptide being an amide - or
carboxy-terminated peptide represented by the following structural formula, and the numbers below each amino acid residue refer to the position of the residue in the peptide:
Leu Lys Leu Leu Lys Lys Leu Leu Lys Lys Leu
1 2 3 4 5 6 7 8 9 10 11
Lys Lys Leu Leu Lys Lys Leu
12 13 14 15 16 17 18
or an analogue of said peptide wherein at least one of amino acid residues 1 through 7, 9, 11, 12, 14, 16, or 18 is deleted from the peptide;
(q) a peptide including the following structural formula X62:
R41-R41-R42-R41-R41-R42-R42-R41-R41-R42-R42-R41-R42-R42, wherein R41 is a hydrophobic amino acid, and R42 is a basic hydrophilic or neutral hydrophilic amino acid;
(r) a peptide including the following structural formula X64:
R42-R41-R41-R42-R42-R41-R42-R42-R41-R41-R42-R42-R41-R41, wherein R41 is a hydrophobic amino acid, and R42 is a basic hydrophilic or neutral hydrophilic amino acid;
(s) a peptide including the following structural formula X66:
R41-R41-R42-R42-R41-R42-R42-R41-R41-R42-R42-R41-R41-R41, wherein R41 is a hydrophobic amino acid, and R42 is a basic
hydrophilic or neutral hydrophilic amino acid;
(t) melittin;
(u) apidaecins;
(v) defensins;
(w) major basic protein of eosinophils;
(x) bacterial permeability-increasing protein; and
(y) perforin.
33. The composition of Claim 32 wherein the peptide is a magainin peptide.
34. The composition of Claim 32 wherein the peptide is a PGLa peptide.
35. The composition of Claim 32 wherein the peptide is an XPF peptide.
36. The composition of Claim 32 wherein the peptide is a CPF peptide.
37. The composition of Claim 32 wherein the peptide is a cecropin.
38. The composition of Claim 32 wherein the peptide is a sarcotoxin.
39. The composition of Claim 32 wherein the peptide includes on of the following basic structures X31 through X37, wherein:
X31 is -[R31-R32-R32-R33-R31-R32-R32]n-;
X32 is -[R32-R32-R32-R33-R31-R32-R32]n-;
X33 is -[R32-R33-R31-R32-R31-R31-R32]n-;
X34 is -[R33-R31-R32-R32-R31-R32-R32]n-;
X35 is -[R31-R32-R32-R31-R32-R32-R33]n-;
X36 is -[R32-R32-R31-R32-R32-R33-R31]n-; and
X37 is -[R32-R31-R32-R32-R33-R31-R32]n-; wherein 31 is basic hydrophilic amino acid, R32 is a hydrophobic amino acid, R33 is a neutral hydrophilic, basic hydrophilic, or hydrophobic amino acid, ar n is from 2 to 5.
40. The composition of Claim 32 wherein the peptide includes
Figure imgf000123_0001
following basic structure X40: R31-R32-R32-R33-R34-R32-R32-R31-R32-R32-R32-R34-R32-R32, wherein R31 is a basic hydrophilic amino acid, R32 is a hydrophobic amino acid, R33 is a neutral hydrophilic or hydrophobic amino acid, and R34 is a basic hydrophilic or hydrophobic amino acid.
41. The composition of Claim 32 wherein said peptide includes the following basic structure X50:
R41-R42-R42-R41-R42-R42-R41-R41-R42-R41-R41,
wherein R41 is a hydrophobic amino acid, and R42 is a basic
hydrophilic or neutral hydrophilic amino acid.
42. The composition of Claim 32 wherein said peptide includes the following basic structure X52:
R42-R41-R42-R42-R41-R41-R42-R42-R41-R42-R42,
wherein R41 is a hydrophobic amino acid, and R42 is a basic
hydrophilic or neutral hydrophilic amino acid.
43. The composition of Claim 32 wherein the peptide includes the following basic structure X54:
-R41-R42-R42-R41-R41-R42-R42-R41 R42-R42-R41-R41-R42-R42-R43-, whereir R41 is a hydrophobic amino acid, R42 is a basic hydrophilic or neutra hydrophilic amino acid, and R43 is a neutral hydrophilic amino acid.
44. The composition of Claim 32 wherein the peptide includes the following basic structure X56:
-R41-R42-R41-R41-R42-R42-R41-R41 R42-R42-R44- ,wherein R41 is a hydrophobic amino acid, R42 is a basic hydrophilic or neutral
hydrophilic amino acid, and R44 is a neutral hydrophilic amino acid o proline.
45. The composition of Claim 32 wherein the peptide includes
Figure imgf000124_0001
following basic structure X58:
-R41-R41-R42-R42-R41-R42-R42-R41-R41-R42-R42-R41-R43-, wherein R41 is a hydrophobic amino acid, R42 is a basic hydrophilic or neutral hydrophilic amino acid, and R43 is a neutral hydrophilic amino acid.
46. The composition of Claim 32 wherein the peptide includes
Figure imgf000124_0002
following basic structure X60:
-R41-R41-R43-R42-R41-R41-R41-R41-R41-R41-R42-R41-R41-R42-R42-R41- R41-R42-R42-R42-R41-, therein R41 is a hydrophobic amino acid, R42 is a basic hydrophilic or neutral hydrophilic amino acid, and R43 is a neutral hydrophilic amino acid.
47. The composition of Claim 32 wherein the peptide has a structure selected from the group consisting of:
(i) R41-R42-R42-R41-R42-R42-R41;
(ii) R41-R41-R42-R42-R41-R42-R42-R41;
(iii) R42-R41-R41-R42-R42-R41-R42-R42-R41;
(iv) R42-R42-R41-R41-R42-R42-R41-R42-R42-R41; and
(v) R41-R42-R42-R41-R41-R42-R42-R41-R42-R42-R41, wherein R41 is a hydrophobic amino acid, and R42 is a basic hydrophilic aminc acid or a neutral hydrophilic amino acid.
48. The composition of Claim 32 wherein said peptide is an amide - or carboxy-terminated peptide represented by the following structural formula, and the numbers below each amino acid residue refer to the position of the residue in the peptide:
Leu Lys Leu Leu Lys Lys Leu Leu Lys Lys Leu
1 2 3 4 5 6 7 8 9 10 11
Lys Lys Leu Leu Lys Lys Leu
12 13 14 15 16 17 18
or an analogue of said peptide wherein at least one of amino acid residues 1 through 7, 9, 11, 12, 14, 16, or 18 is deleted from the peptide.
49. The process of Claim 32 wherein said peptide includes the following structural formula X62:
-R41-R41-R42-R41-R41-R42-R41-R41 R41-R42-R41-R41-R42-R42-R42-, wherein R41 is a hydrophobic amino acid, and R41 is a basic
hydrophilic or neutral hydrophilic amino acid.
50. The process of Claim 32 wherein said peptide includes the following structural formula X64:
-R42-R41-R41-R42-R42-R41-R42-R42 R41-R41-R42-R42-R41-R41, wherein R41 is a hydrophobic amino acid, and R42 is a basic
hydrophilic or neutral hydrophilic amino acid.
51. The process of Claim 32 wherein said peptide includes the following structural formula X66: -R41-R41-R42-R42-R41-R42-R42-R41-R41-R42-R42-R41-R41,
wherein R41 is a hydrophobic amino acid, and R42 is a basic
hydrophilic or neutral hydrophilic amino acid.
52. The composition of Claim 32 wherein said peptide is
melittin.
53. The composition of Claim 32 wherein said peptide is an apidaecin.
54. The composition of Claim 32 wherein said peptide or protein is a defensin.
55. The composition of Claim 32 wherein said peptide or protein is major basic protein of eosinophils.
56. The composition of Claim 32 wherein said peptide or protein is bacterial permeability-increasing protein.
57. The composition of Claim 32 wherein said pjeptide or protein is perforin.
58. The composition of Claim 31 wherein said at least one protease inhibitor is bestatin.
59. The composition of Claim 31 wherein each of components (a) and (b) is present in an amount ineffective in treating a cancerous growth if administered alone to a host.
PCT/US1994/002121 1993-02-26 1994-02-22 Treatment of cancerous growths with biologically active peptides and protease inhibitors WO1994019369A1 (en)

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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1996028468A2 (en) * 1995-03-09 1996-09-19 Unilever Plc Amphiphilic peptide and analogs thereof
EP0932420A1 (en) * 1996-04-30 1999-08-04 President And Fellows Of Harvard College Drug delivery using terminal complement components
US6440690B1 (en) 1995-06-29 2002-08-27 Amram Mor Peptides for the activation of the immune system in humans and animals
US8318899B2 (en) 2008-01-24 2012-11-27 Board Of Supervisors Of Louisiana State University And Agricultural And Mechanical College Lytic domain fusion constructs and methods of making and using same
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Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1996028468A2 (en) * 1995-03-09 1996-09-19 Unilever Plc Amphiphilic peptide and analogs thereof
WO1996028468A3 (en) * 1995-03-09 1997-04-17 Unilever Plc Amphiphilic peptide and analogs thereof
US6440690B1 (en) 1995-06-29 2002-08-27 Amram Mor Peptides for the activation of the immune system in humans and animals
EP0932420A1 (en) * 1996-04-30 1999-08-04 President And Fellows Of Harvard College Drug delivery using terminal complement components
EP0932420A4 (en) * 1996-04-30 2000-05-31 Harvard College SUPPLY OF MEDICINAL PRODUCTS BY MEANS OF COMPLEMENT TERMINALS
US8318899B2 (en) 2008-01-24 2012-11-27 Board Of Supervisors Of Louisiana State University And Agricultural And Mechanical College Lytic domain fusion constructs and methods of making and using same
US8546535B2 (en) 2008-01-24 2013-10-01 Esperance Pharmaceuticals, Inc. Lytic domain fusion constructs and methods of making and using same
US9255134B2 (en) 2008-01-24 2016-02-09 Esperance Pharmaceuticals, Inc. Lytic domain fusion constructs and methods of making and using same
US9492563B2 (en) 2012-10-30 2016-11-15 Esperance Pharmaceuticals, Inc. Antibody/drug conjugates and methods of use
US10233214B2 (en) 2012-10-30 2019-03-19 Esperance Pharmaceuticals, Inc. Antibody/drug conjugates and methods of use

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