[go: up one dir, main page]

WO1994019369A1 - Traitement de tumeurs cancereuses au moyen d'inhibiteurs de protease et de peptides a activite biologique - Google Patents

Traitement de tumeurs cancereuses au moyen d'inhibiteurs de protease et de peptides a activite biologique Download PDF

Info

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
Authority
WO
WIPO (PCT)
Prior art keywords
amino acid
peptide
lys
hydrophilic
ala
Prior art date
Application number
PCT/US1994/002121
Other languages
English (en)
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/fr

Links

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

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Epidemiology (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Immunology (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Veterinary Medicine (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Zoology (AREA)
  • Marine Sciences & Fisheries (AREA)
  • Molecular Biology (AREA)
  • Peptides Or Proteins (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

Des compositions comprenant des inhibiteurs de protéase et des peptides à activité biologique se prêtent à une utilisation comme produits pharmaceutiques dans le traitement du cancer.
PCT/US1994/002121 1993-02-26 1994-02-22 Traitement de tumeurs cancereuses au moyen d'inhibiteurs de protease et de peptides a activite biologique WO1994019369A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU62515/94A AU6251594A (en) 1993-02-26 1994-02-22 Treatment of cancerous growths with biologically active peptides and protease inhibitors

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US2160793A 1993-02-26 1993-02-26
US08/021,607 1993-02-26

Publications (1)

Publication Number Publication Date
WO1994019369A1 true WO1994019369A1 (fr) 1994-09-01

Family

ID=21805153

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1994/002121 WO1994019369A1 (fr) 1993-02-26 1994-02-22 Traitement de tumeurs cancereuses au moyen d'inhibiteurs de protease et de peptides a activite biologique

Country Status (2)

Country Link
AU (1) AU6251594A (fr)
WO (1) WO1994019369A1 (fr)

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1996028468A3 (fr) * 1995-03-09 1997-04-17 Unilever Plc Peptides amphiphiles et analogues desdits peptides
EP0932420A4 (fr) * 1996-04-30 2000-05-31 Harvard College Apport de medicaments au moyen de constituants terminaux de complement
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
US9492563B2 (en) 2012-10-30 2016-11-15 Esperance Pharmaceuticals, Inc. Antibody/drug conjugates and methods of use

Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4029547A (en) * 1974-07-01 1977-06-14 Zaidan Hojin Biseibutsu Kagaku Kenkyu Kai Biologically active substance, bestatin, and production thereof
US4052449A (en) * 1974-07-01 1977-10-04 Zaidan Hojin Biseibutsu Kagaku Kenkyu Kai Biologically active substance, bestatin, and production thereof
US4179573A (en) * 1974-07-01 1979-12-18 Zaidan Hojin Biseibutsu Kagaku Kenkyu Kai (2S,3R)-3-amino-2-hydroxy-4-phenylbutanoic acid
US5045531A (en) * 1989-12-18 1991-09-03 Magainin Sciences Inc. Wound treatment employing biologically active ion channel forming peptides and proteins
US5106948A (en) * 1988-05-27 1992-04-21 Mao Foundation For Medical Education And Research Cytotoxic boronic acid peptide analogs
US5114921A (en) * 1988-05-27 1992-05-19 The Children's Hospital Of Philadelphia Amphiphilic peptides and use thereof
US5159060A (en) * 1988-05-27 1992-10-27 Mayo Foundation For Medical Education And Research Cytotoxic boronic acid peptide analogs
US5217956A (en) * 1988-10-21 1993-06-08 The Children's Hospital Of Philadelphia Composition and treatment with biologically active peptides and certain anions
US5254535A (en) * 1989-04-17 1993-10-19 The Children's Hospital Of Pennsylvania Composition and treatment with biologically active peptides and antibiotic

Patent Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4029547A (en) * 1974-07-01 1977-06-14 Zaidan Hojin Biseibutsu Kagaku Kenkyu Kai Biologically active substance, bestatin, and production thereof
US4052449A (en) * 1974-07-01 1977-10-04 Zaidan Hojin Biseibutsu Kagaku Kenkyu Kai Biologically active substance, bestatin, and production thereof
US4179573A (en) * 1974-07-01 1979-12-18 Zaidan Hojin Biseibutsu Kagaku Kenkyu Kai (2S,3R)-3-amino-2-hydroxy-4-phenylbutanoic acid
US5106948A (en) * 1988-05-27 1992-04-21 Mao Foundation For Medical Education And Research Cytotoxic boronic acid peptide analogs
US5114921A (en) * 1988-05-27 1992-05-19 The Children's Hospital Of Philadelphia Amphiphilic peptides and use thereof
US5159060A (en) * 1988-05-27 1992-10-27 Mayo Foundation For Medical Education And Research Cytotoxic boronic acid peptide analogs
US5217956A (en) * 1988-10-21 1993-06-08 The Children's Hospital Of Philadelphia Composition and treatment with biologically active peptides and certain anions
US5254535A (en) * 1989-04-17 1993-10-19 The Children's Hospital Of Pennsylvania Composition and treatment with biologically active peptides and antibiotic
US5045531A (en) * 1989-12-18 1991-09-03 Magainin Sciences Inc. Wound treatment employing biologically active ion channel forming peptides and proteins

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1996028468A3 (fr) * 1995-03-09 1997-04-17 Unilever Plc Peptides amphiphiles et analogues desdits peptides
US6440690B1 (en) 1995-06-29 2002-08-27 Amram Mor Peptides for the activation of the immune system in humans and animals
EP0932420A4 (fr) * 1996-04-30 2000-05-31 Harvard College Apport de medicaments au moyen de constituants terminaux de complement
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

Also Published As

Publication number Publication date
AU6251594A (en) 1994-09-14

Similar Documents

Publication Publication Date Title
USRE35492E (en) Wound treatment employing biologically active ion channel forming peptides and proteins
EP0809655A1 (fr) PEPTIDES ET MOLECULES A COMPORTEMENT MIMETIQUE DES PEPTIDES AYANT UNE SIMILARITE STRUCTURELLE AVEC LA PROTEINE p53 HUMAINE ACTIVANT LA FONCTION p53
AU674525B2 (en) Biologically active peptides having N-terminal substitutions
WO1994012206A1 (fr) Traitement de tumeurs dermatologiques malignes avec des peptides bioactifs
US5635479A (en) Treatment of gynecological malignancies with biologically active peptides
AU5741794A (en) Treatment of septic shock with conjugated biologically active peptides
AU3236293A (en) Composition and treatment with biologically active peptides and chelating agents
EP0590044A4 (en) Composition and treatment with biologically active peptides having c-terminal substitutions
AU2323792A (en) Treating the oral cavity with ion-channel forming peptides
WO1994019369A1 (fr) Traitement de tumeurs cancereuses au moyen d'inhibiteurs de protease et de peptides a activite biologique
AU641129B2 (en) Composition and treatment with biologically active peptides and certain anions
AU693518B2 (en) Ion-channel forming amphiphilic peptides having n-terminal modifications
WO1991016918A1 (fr) Composition et traitement utilisant des peptides biologiquement actifs ainsi que des agents parasiticides et des agents fongicides
EP1001800A2 (fr) Peptides biologiquement actifs a toxicite reduite chez l'animal et procedes d'elaboration

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AU CA JP

AL Designated countries for regional patents

Kind code of ref document: A1

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

121 Ep: the epo has been informed by wipo that ep was designated in this application
DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
122 Ep: pct application non-entry in european phase
NENP Non-entry into the national phase

Ref country code: CA