+

WO2009070564A2 - Compositions et procédés utilisables dans le cadre de la prévention d'infections associées à des implants transcutanés ostéointégrés - Google Patents

Compositions et procédés utilisables dans le cadre de la prévention d'infections associées à des implants transcutanés ostéointégrés Download PDF

Info

Publication number
WO2009070564A2
WO2009070564A2 PCT/US2008/084640 US2008084640W WO2009070564A2 WO 2009070564 A2 WO2009070564 A2 WO 2009070564A2 US 2008084640 W US2008084640 W US 2008084640W WO 2009070564 A2 WO2009070564 A2 WO 2009070564A2
Authority
WO
WIPO (PCT)
Prior art keywords
peptide
lys
wound
amino acids
amino acid
Prior art date
Application number
PCT/US2008/084640
Other languages
English (en)
Other versions
WO2009070564A3 (fr
Inventor
Roy D. Bloebaum
Cathy Petti
Juliana Szakacs
Teri G.R. Chou
Original Assignee
Macrochem Corporation
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 Macrochem Corporation filed Critical Macrochem Corporation
Publication of WO2009070564A2 publication Critical patent/WO2009070564A2/fr
Publication of WO2009070564A3 publication Critical patent/WO2009070564A3/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
    • A61K38/16—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
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics

Definitions

  • This invention relates to the use of peptides in preventing and treating infections. More specifically, the invention relates to the use of amphiphilic peptides in preventing and treating infections. Even more specifically the invention relates to the use of cationic amphiphilic peptides in preventing and treating infections associated with transcutaneous osseointegreated implants.
  • socket technology pain from loading soft tissue with underlying heterotopic ossification (Potter BK, Burns TC, Lacap AP, Granville RR, Gajewski D. Heterotopic ossification in the residual limbs of traumatic and combat-related amputees. Journal of the American Academy of Orthopaedic Surgeons. 2006 Oct;14(10 Suppl):S191-7) and soft tissue atrophy (Fernie GR, Holliday PJ. Volume fluctuations in the residual limbs of lower limb amputees. Arch Phys Med
  • Bone is the major biological tissue designed to carry the biomechanical loads associated with motion in the human body. Therefore, the ideal strategy for rehabilitating the amputee is to develop skeletal implants that are maintained without infection and remain attached to the human skeleton for the lifetime of the amputee.
  • Branemark recently demonstrated that osseointegrated implants affixed to the skeleton can substantially improve the quality of life for amputee patients (Hagberg K. Transfemoral amputation, quality of life and prosthetic function. Goteborg, Sweden: The Sahlgrenska Academy at Goteborg University; 2006) as they avoid the limitations associated with socket technology.
  • antimicrobial peptides may be useful in preventing wound infection.
  • Ge Y MacDonald DL, Holroyd KJ, Thornsberry C, Wexler H, Zasloff M. In vitro antibacterial properties of pexiganan, an analog of magainin.
  • Antimicrobial peptides including amphiphilic peptides, are used by animals as part of their innate immunity against microbes in their environment (Boman HG.
  • Antibacterial peptides basic facts and emerging concepts. J Intern Med. 2003 Sep;254(3):197-215; Boman HG. Peptide antibiotics and their role in innate immunity. Annu Rev Immunol. 1995;13:61-92; Schonwetter BS, Stolzenberg ED, Zasloff MA. Epithelial antibiotics induced at sites of inflammation. Science. 1995 Mar 17;267(5204):1645-8; Stolzenberg ED, Anderson GM, Ackermann MR, Whitlock RH,
  • proline-arginine-rich peptides (Agerberth et al., Eur. J. Biochem. 202, 849 854 (1991)), and sapecin (Matsuyama et al., J. Biol. Chem. 263, 17112 17116 (1988)), like beta peptides, all have a net positive charge and considerable hydrophobic character.
  • magainins isolated from the skin of the African clawed frog ⁇ Xenopus laevis
  • Iwahori A Hirota Y, Sampe R, Miyano S, Numao N. Synthesis of reversed magainin 2 analogs enhanced antibacterial activity. Biol Pharm Bull. 1997 Mar;20(3):267-70;
  • Magainin 2 a 23-amino acid amphiphilic cationic peptide
  • pathogens including bacteria, amoebae, fungi, and parasites.
  • antibiotic resistance to the magainin family of peptides has been no documented antibiotic resistance to the magainin family of peptides (Ge Y, MacDonald DL, Holroyd KJ, Thornsberry C, Wexler H, Zasloff M. In vitro antibacterial properties of pexiganan, an analog of magainin. Antimicrob Agents Chemother. 1999 Apr;43(4):782-8; Hancock RE. Peptide antibiotics. Lancet.
  • This invention comprises a method of preventing infections using amphiphilic peptides. More specifically, the invention relates to the use of cationic amphiphilic peptides in preventing infections associated with transcutaneous implants.
  • the cationic amphiphilic peptide is a mangainin peptide or mangainin peptide derivative.
  • mangainin peptide is pexiganan or derivative thereof.
  • the invention comprises a method of preventing wound infection by the topical application of a cationic peptide to a wound site, hi one embodiments the cationic peptide is a mangainin.
  • the mangainin peptide is pexiganan or a analog therefore.
  • the pexiganan is an acid, amid or salt of the peptide shown in SEQ ID NO: 1.
  • the invention comprises a process for preventing wound infection, comprising topically administering a therapeutic formulation of a magainin peptide having the following peptide structure:
  • the invention comprises a process for preventing wound infection, comprising topically administering a therapeutic formulation of a magainin peptide having the following peptide structure:
  • the invention comprises a process for preventing would infection by administering topically a therapeutic formulation comprising a cationic peptide wherein the peptide has the amino actid sequence of SEQ ID NO: 1.
  • the invention comprises a method of preventing wound infection by the topical application of a therapeutic formulation comprising a cream, the cream further comprising a cationic peptide to a wound site.
  • the peptide is a salt, amide, or acid of pexiganan.
  • the therapeutic formulation comprises a 1% pexiganan acetate cream.
  • the therapeutic formulation is applied to the wound site once a day.
  • the invention comprises a process for preventing wound infection in a host, comprising administering to a host having a wound at least one biologically active amphiphilic peptide and/or biologically active protein, said peptide or protein being an ion channel-forming peptide or protein, said at least one biologically active amphiphilic peptide or protein being administered in an amount effective for preventing wound infection in the host.
  • the peptide of the process of the invention is a basic polypeptide having at least sixteen amino acids, wherein said basic polypeptide includes at least eight hydrophobic amino acids and at least eight hydrophilic amino acids.
  • the amphiphilic peptide is cationic. More preferably the peptide is a mangainin peptide.
  • the invention comprises a process for preventing wound infection in a host, comprising administering to a host having a wound a magainin peptide having the following basic peptide structure:
  • the invention comprises a process for preventing wound infection in a host, comprising administering to a host having a wound a magainin peptide having the following basic peptide structure:
  • Rn is a hydrophobic amino aid
  • Ri 2 is a basic hydrophilic amino acid
  • Rn is a hydrophobic, neutral hydrophilic, or basic hydrophilic amino acid
  • Ri 4 and Rj 43 are hydrophobic or basic hydrophilic amino acids.
  • the invention comprises a process for preventing wound infection in a host, comprising administering to a host having a wound at least one biologically active amphiphilic peptide and/or biologically active protein, wherein the wound is a transcutaneous wound.
  • the transcutaneous wouldn is the site of a medical implant.
  • the invention comprises a process for preventing wound infection in a host, comprising administering to a host having a wound at least one biologically active amphiphilic peptide and/or biologically active protein wherein the peptide has the following peptide structure:
  • FIGURE 1 is a depiction of the surgery involved in inserting transcutaneous implants.
  • the implant went through the tibialis anterior muscle and remained transcutaneous on the lateral side.
  • the ends of the cerclage wire were twisted together, trimmed, and pressed flat against the medical side of the tibia underneath the soft tissue.
  • FIGURE 2 is a scanning electron image of a tantalum implant depicting the area analyzed for bone ingrowth.
  • FIGURE 3 shows that certain porous tantalum implants used in the procedures of the examples were prone to breaking and to skin healing over the implant, thereby, sealing the implant from the external environment.
  • the rabbits were removed from the study since they no longer had a pathway for pin track infection.
  • compositions of the present invention include one or more polypeptides.
  • Amino acid is used herein to refer to a chemical compound with the general formula: NH 2 -CRH-COOH, where R, the side chain, is H or an organic group. Where R is an organic group, R can vary and is either polar or nonpolar (i.e., hydrophobic).
  • the amino acids of this invention can be naturally occurring or synthetic (often referred to as nonproteinogenic).
  • an organic group is a hydrocarbon group that is classified as an aliphatic group, a cyclic group or combination of aliphatic and cyclic groups.
  • aliphatic group means a saturated or unsaturated linear or branched hydrocarbon group. This term is used to encompass alkyl, alkenyl, and alkynyl groups, for example.
  • cyclic group means a closed ring hydrocarbon group that is classified as an alicyclic group, aromatic group, or heterocyclic group.
  • alicyclic group means a cyclic hydrocarbon group having properties resembling those of aliphatic groups.
  • aromatic group refers to mono- or polycyclic aromatic hydrocarbon groups. As used herein, an organic group can be substituted or unsubstituted.
  • polypeptide and peptide are used interchangeably and refer to a polymer of amino acids. These terms do not connote a specific length of a polymer of amino acids. Thus, for example, the terms oligopeptide, protein, and enzyme are included within the definition of polypeptide or peptide, whether produced using recombinant techniques, chemical or enzymatic synthesis, or naturally occurring. This term also includes polypeptides that have been modified or derivatized, such as by glycosylation, acetylation, phosphorylation, and the like.
  • Cys Cysteine
  • Leu Leucine
  • bacteria preferably Gram-negative bacteria
  • the types of bacteria can include Pseudomonas spp including P. aeruginosa and P. cepacia, E. coli strains, including E. coli B, Salmonella, Proteus mirabilis and Staphylococcus strains such as Staphylococcus aureus.
  • a preferred organism is Staphylococcus aureus.
  • polypeptides of of the invention can be in their free acid form or they can be amidated at the C-terminal carboxylate group.
  • the present invention also includes analogs of the polypeptide of the invetion, which typically have structural similarity with the diclsosed polypeptides.
  • An "analog" of a polypeptide includes at least a portion of the polypeptide, wherein the portion contains deletions or additions of one or more contiguous or noncontiguous amino acids, or containing one or more amino acid substitutions. Substitutes for an amino acid in the polypeptides of the invention are preferably conservative substitutions, which are selected from other members of the class to which the amino acid belongs.
  • an amino acid belonging to a grouping of amino acids having a particular size or characteristic can generally be substituted for another amino acid without substantially altering the structure of a polypeptide.
  • conservative amino acid substitutions are defined to result from exchange of amino acids residues from within one of the following classes of residues: Class I: Ala, GIy, Ser, Thr, and Pro (representing small aliphatic side chains and hydroxyl group side chains); Class II: Cys, Ser, Thr, and Tyr (representing side chains including an -OH or -SH group); Class III: GIu, Asp, Asn, and GIn (carboxyl group containing side chains): Class IV: His, Arg, and Lys
  • Class V He, VaI, Leu, Phe, and Met (representing hydrophobic side chains); and Class VI: Phe, Trp, Tyr, and His (representing aromatic side chains).
  • the classes also include related amino acids such as 3Hyp and 4Hyp in Class I; homocysteine in Class II; 2-aminoadipic acid, 2-aminopimelic acid, .gamma.- carboxyglutamic acid, .beta.-carboxyaspartic acid, and the corresponding amino acid amides in Class III; ornithine, homoarginine, N-methyl lysine, dimethyl lysine, trimethyl lysine, 2,3-diaminopropionic acid, 2,4-diaminobutyric acid, homoarginine, sarcosine and hydroxylysine in Class IV; substituted phenylalanines, norleucine, norvaline, 2- aminooctanoic acid, 2-amin
  • Polypeptide analogs as that term is used herein, also include modified polypeptides. Modifications of polypeptides of the invention include chemical and/or enzymatic derivatizations at one or more constituent amino acid, including side chain modifications, backbone modifications, and N- and C-terminal modifications including acetylation, hydroxylation, methylation, amidation, and the attachment of carbohydrate or lipid moieties, cofactors, and the like.
  • a preferred polypeptide analog is characterized by having at least one of the biological activities described herein. Such an analog is referred to herein as a "biologically active analog" or simply “active analog.”
  • the biological activity of a polypeptide can be determined, for example, as described in the Examples Section.
  • the polypeptides of the invention may be synthesized by the solid phase method using standard methods based on either t-butyloxycarbonyl (BOC) or 9- fluorenylmethoxy-carbonyl (FMOC) protecting groups. This methodology is described by G. B. Fields et al. in Synthetic Peptides: A User's Guide, W.M. Freeman & Company, New York, N.Y., pp. 77 183 (1992).
  • the present peptides may also be synthesized via recombinant techniques well known to those skilled in the art. For example, U.S. Pat.
  • No. 5,595,887 describes methods of forming a variety of relatively small peptides through expression of a recombinant gene construct coding for a fusion protein which includes a binding protein and one or more copies of the desired target peptide. After expression, the fusion protein is isolated and cleaved using chemical and/or enzymatic methods to produce the desired target peptide.
  • the biologically active peptides or biologically active proteins employed in the present invention are ion channel-forming peptides or proteins.
  • 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 P. 5072-76 (July, 1988) describes methodology which indicates whether or not a peptide or protein has ion channel-forming properties and is therefore an ionophore.
  • an ion channel-forming peptide or protein is a peptide or protein which has ion channel-forming properties as determined by the method of Christensen et al.
  • the peptides employed in this invention are preferably amphilic peptides and more preferably amphiphilic acationic peptides.
  • An amphiphilic peptide is a peptide which includes both hydrophobic and hydrophilic peptide regions.
  • a cationic peptide is one that carries a positive charge.
  • the administration of the biologically active amphiphilic peptides or proteins to a host may is preferably by topical administration in a consistent and repeated manner in order to prvent infention of the host.
  • the topical administration is prefarably performed at or near a wound site where infection would tend to occur.
  • the peptide employed, or derivatives or analogues thereof 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.
  • 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.
  • the polypeptide comprises a chain of at least four groups 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, GIy, He, Leu, Met, VaI, Trp, and Tyr.
  • the neutral hydrophilic amino acids may be selected from the class consisting of Asn, GIn, Ser, and Thr.
  • the basic hydrophilic amino acids may be selected from the class consisting of Lys, Arg, His, and ornithine (O).
  • 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
  • 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 comprises 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, 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 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: (XO a (A-B-C-D) n (YO b
  • Yi 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 is -C, -C-D, -C-D-A
  • X 4 is C-, B-C-, A-B-C-
  • Y 4 is D, -D-A, -D-A-B a is 0 or l; b is O or 1 and 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 the chain.
  • the chains may have a Ser-Lys sequence before the "Ala” end, and/or an AIa- Phe sequence after the "Lys" end.
  • Other amino acid sequences may also be attached to the "Ala” and/or the "Lys" end.
  • 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.
  • the peptides may be produced by known techniques and obtained in substantially pure form. For example, the peptides may be synthesized on an automatic synthesizer. Journal of the American Chemical Society, Vol. 85 Pages 2149-54(1963). It is also possible to produce such peptides by genetic engineering techniques.
  • the peptide employed 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 may include the following basic peptide structure X 12 :
  • Rn is a hydrophobic amino acid
  • Rj 2 is a basic hydrophilic amino acid
  • Rn is a hydrophobic, neutral hydrophilic, or basic hydrophilic amino acid
  • Ri 4 and Ri 4a are hydrophobic or basic hydrophilic amino acids
  • Ri 5 is glutamic acid or aspartic acid, or a hydrophobic or basic hydrophilic amino aid
  • n is 0 or 1.
  • Rn is a hydrophobic or neutral hydrophilic amino acid
  • Ri 4a 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 17 is a neutral hydrophilic amino acid, a hydrophobic amino acid, or a basic hydrophilic amino acid.
  • R] 7 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
  • the magainin peptide may also include the following structure Xi 3 -Zi 3 ; wherein X] 3 is the herein above described basic peptide structure and Zi 3 is:
  • Rn, R 14 , R 14a , R ]5 , Ri 6 , and Rn are amino acids as hereinabove described, and n is 0 or 1, and each n may be the same or different.
  • the peptide of the invention is a mangainin peptide or anlaog thereof.
  • the magainin peptides generally 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 are described in Proc. Natl. Acad. Sci. Vol. 84 pp. 5449-53 (Aug. 1987).
  • magainin peptides refers to the basic magainin structure as well as derivatives and analogs thereof, including but not limited to the representatives derivatives or analogs disclosed herein.
  • magainin peptides having the following primary sequence (expressed as a single letter code) as well as appropriate analogues and derivatives thereof:
  • the present invention also provides a composition that includes one or more active agents (i.e., polypeptides) of the invention and one or more pharmaceutically acceptable carriers.
  • active agents i.e., polypeptides
  • One or more polypeptides with demonstrated biological activity can be administered to a patient in an amount alone or together with other active agents and with a pharmaceutically acceptable buffer.
  • the polypeptides can be combined with a variety of physiological acceptable carriers for delivery to a patient including a variety of diluents or excipients known to those of ordinary skill in the art. For example, for parenteral administration, isotonic saline is preferred.
  • a cream including a carrier such as dimethylsulfoxide (DMSO), or other agents typically found in topical creams that do not block or inhibit activity of the peptide, can be used.
  • a carrier such as dimethylsulfoxide (DMSO)
  • DMSO dimethylsulfoxide
  • suitable carriers include, but are not limited to, alcohol, phosphate buffered saline, and other balanced salt solutions.
  • the formulations may be conveniently presented in unit dosage form and may be prepared by any of the methods well known in the art of pharmacy. Preferably, such methods include the step of bringing the active agent into association with a carrier which constitutes one or more accessory ingredients.
  • the formulations are prepared by uniformly and intimately bringing the active agent into association with a liquid carrier, a finely divided solid carrier, or both, and then, if necessary, shaping the product into the desired formulations.
  • the methods of the invention include administering to a patient, preferably a mammal, and more preferably a human, the composition of the invention in an amount effective to produce the desired effect.
  • the peptides can be administered as a single dose or in multiple doses.
  • Useful dosages of the active agents can be determined by comparing their in vitro activity and the in vivo activity in animal models. Methods for extrapolation of effective dosages in mice, and other animals, to humans are known in the art; for example, see U.S. Pat. No. 4,938,949.
  • the agents of the present invention are preferably formulated in pharmaceutical compositions and then, in accordance with the methods of the invention, administered to a patient, such as a human patient, in a variety of forms adapted to the chosen route of administration.
  • the formulations include, but are not limited to, those suitable for oral, rectal, vaginal, topical, nasal, ophthalmic, or parental (including subcutaneous, intramuscular, intraperitoneal, intratumoral, and intravenous) administration.
  • Formulations suitable for parenteral administration conveniently include a sterile aqueous preparation of the active agent, or dispersions of sterile powders of the active agent, which are preferably isotonic with the blood of the recipient.
  • Isotonic agents that can be included in the liquid preparation include sugars, buffers, and sodium chloride.
  • Solutions of the active agent can be prepared in water, optionally mixed with a nontoxic surfactant.
  • Dispersions of the active agent can be prepared in water, ethanol, a polyol (such as glycerol, propylene glycol, liquid polyethylene glycols, and the like), vegetable oils, glycerol esters, and mixtures thereof.
  • the ultimate dosage form is sterile, fluid, and stable under the conditions of manufacture and storage.
  • the necessary fluidity can be achieved, for example, by using liposomes, by employing the appropriate particle size in the case of dispersions, or by using surfactants.
  • Sterilization of a liquid preparation can be achieved by any convenient method that preserves the bioactivity of the active agent, preferably by filter sterilization.
  • Preferred methods for preparing powders include vacuum drying and freeze drying of the sterile injectible solutions. Subsequent microbial contamination can be prevented using various antimicrobial agents, for example, antibacterial, antiviral and antifungal agents including parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. Absorption of the active agents over a prolonged period can be achieved by including agents for delaying, for example, aluminum monostearate and gelatin.
  • antimicrobial agents for example, antibacterial, antiviral and antifungal agents including parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like.
  • Absorption of the active agents over a prolonged period can be achieved by including agents for delaying, for example, aluminum monostearate and gelatin.
  • the peptide or protein is employed to provide peptide or protein dosages of from 0.1 mg to 250 mg per kilogram of host weight, when administered systemically.
  • the peptide or protein is administered in an amount of from about 0.1% to about 10%, preferably from about 0.5% to about 2%.
  • the topical composition may be in the form of an ointment, cream, or solution.
  • the composition is a topical cream or ointment comprising about 1% pexiganan or its derivatives, aanalogs, salts or amids.
  • the implants were 36-mm long and 2.7-mm diameter. A hexagonal cross- section was used to prevent rotational motion.
  • the implants were either titanium alloy
  • Ti-6A1-4V free of surface finish or tantalum with uniform volume porosity of 75% to 80%
  • Trabecular MetalTM Zimmer, Inc., Warsaw, IN
  • All implants were manufactured by the Zimmer Corporation.
  • the implants were pre-sterilized by gamma radiation and conformed to Zimmer's human implant sterile lot release procedures.
  • the investigators added a less than 1.0-mm diameter hole to the tip of the implant to allow it to be secured the tibia with a cerclage wire.
  • the implants were then passivated by MedicineLodge, Inc (Logan, UT), re-sterilized in an autoclave, and placed in a sterile pouch.
  • aureus consisted of phosphate buffered saline with 25% glycerol. The bacteria solution was frozen until just prior to inoculation. Inoculation was performed by dripping the bacteria solution over the implant site with a sterile transfer pipet.
  • the S. aureus used in this study came from an American Type Culture Collection (ATCC) 49230, which was from a patient with osteomyelitis. Table 1 - Description and sample size of each experimental rabbit group. Group 1 was compared to Group 2 for addressing the first hypothesis, and Group 1 was compared to Group 3 for addressing the second hypothesis.
  • Anesthesia was induced by an intramuscular injection of ketamine (35mg/kg) and Xylazine (5mg/kg) into the rabbit's semi tendonosis or semi membranosis muscle.
  • ketamine 35mg/kg
  • Xylazine 5mg/kg
  • swab samples were obtained at the implant site. After endotracheal intubation the rabbit breathed isoflurane (3-5%) in oxygen and respiration was monitored with an apnea detection device.
  • An alpha 2 antagonist, Yohimine or Tolazine HCL was administered IV (0.5mg/kg) in the lateral ear vein to partially reverse the effect of Xylazine.
  • buprenorphine (0.02-0.1 mg/kg) was injected subcutaneously into the rabbit's neck region.
  • a 25 ⁇ g transdermal fentanyl patch was sutured to the shaved spot in between the rabbit's shoulders. Suturing prevented the patch from coming off during the rabbit's recovery.
  • the hind leg was close-shaved and prepared for surgery by scrubbing with a povidine iodine scrub followed by a 70% alcohol wipe repeated three times. After a brief drying time, the area was prepped with a final spray of povidine solution and again allowed to dry.
  • the rabbit's heart rate, mucus membrane color, corneal reflex, capillary refill time and respiratory rate were monitored and recorded.
  • a direct anterolateral incision (1.0 ⁇ 0.2cm) was made over the proximal tibia while the limb was positioned in extension. The skin and fascia were reflected. The fibers of the tibialis anterior muscle were separated so that the implant traversed the muscle belly.
  • a bicortical hole was drilled in the proximal tibia approximately 1-cm distal to the tibia tuberosity. The implants were tapped into the hole with a metal mallet for press-fit fixation.
  • a 24 gauge cerclage wire was inserted through the hole in the medial end of the implant.
  • One end of the wire was placed along the anterior cortex of the tibia, taken around the shaft of the implant on the lateral side, and then both ends of the wire were twisted together on the medial side.
  • the ends of the wire were trimmed to about 2-mm and pressed flat with needle holders against the tibia ( Figure 1).
  • the rabbits were euthanized as per protocol when they showed clinical signs greater than a Grade I pin track infection but no greater than a Grade II infection (Checketts RG, MacEachern AG, Otterburn M. Pin track infection and the principles of pin site care.
  • a Grade II infection was defined as "slight redness around the pin together will a little discharge” whereas a Grade II infection was defined as the presence of swelling, redness, purulent discharge, and tenderness of the implant site (Checketts RG, MacEachern AG, Otterburn M. Pin track infection and the principles of pin site care.
  • Gia Grade II infection was defined as "slight redness around the pin together will a little discharge”
  • a Grade II infection was defined as the presence of swelling, redness, purulent discharge, and tenderness of the implant site (Checketts RG, MacEachern AG, Otterburn M. Pin track infection and the principles of pin site care.
  • Gia Grade II infection was defined as "slight redness
  • samples were obtained of blood, muscle, and bone. All skin surfaces were disinfected with a povidone-iodine solution and all instruments were sterilized or disinfected prior to tissue collection.
  • blood was drawn from a vein or artery from the inside of the ear and injected into a blood culture bottle. After euthanasia, a biopsy punch was used to collect the muscle samples. This sample extended from the skin surface to the bone.
  • the muscle sample was taken 5 -mm away from the implant site in the direction towards the knee joint. The muscle sample was then placed into fastidious broth for culturing.
  • the soft tissue around the tibia was cut with a scalpel circumferentially 4-cm distal from the implant site.
  • the tibia was transected using an autopsy saw. Bone chips were collected by twisting a 1/8 inch drill bit inside the medullary canal near the implant. The entire drill bit was placed into fastidious broth. After using standard laboratory protocols to culture the tissue samples, the presence or absence of bacteria growth was noted as well as the type of organism such as S. aureus or E. coli. The technician performing the cultures and analysis was not aware of the rabbit study groups the samples belonged.
  • the tibia was disarticulated and placed in 70% ethanol. After taking photos and radiographs of the gross specimens, they were fixed in 1 :10 buffered formalin. Within days, a transverse 3-ram thick slice of the tibia was taken so that the side of the slice closest to the implant was at a distance of 1 to 3-mm. Histology ink was used to mark the side of the section that was furthest from the implant and the lateral side of the skin, muscle, and bone. Skin and muscle were placed into one cassette and the bone in another cassette. Care was taken to maintain the physiological orientation of the tissue.
  • the bone was decalcified and then both the bone and soft tissue were dehydrated, infiltrated (Tissue Tek VAccum Infiltration Process, Miles Scientific), and embedded in paraffin (Histocentre 2, Shandon).
  • a microtome was used to cut 4- ⁇ m slices of the specimens. At least three slices were taken of each soft tissue and bone specimen. These were stained with Hematoxylin & Eosin (H&E) and histologically examined for inflammation, fibrosis, and bone remodeling. Periodic acid- Schiff (PAS) stain was used to detect the presence of fungus, and Brown-Brenn stain was used to detect the presence of bacteria.
  • H&E Hematoxylin & Eosin
  • PAS Periodic acid- Schiff
  • the H&E and PAS stain was performed using a Microm DS 50 Slide Stainer (Richard-Allan Scientific).
  • the Brown-Brenn stains as well as all histological analyses were performed by the Department of Pathology at Harvard Vanguard Medical Associates (Boston, MA). This analysis performed while being blinded to the specimen's study group. Positive histology, indicative of infection, was defined as the presence of acute inflammation and microorganisms on the Brown-Brenn and/or PAS stain.
  • the residual tibia was dehydrated, infiltrated, and embedded in methyl methacylate using standard procedures (Emmanual J, Hornbeck C, Bloebaum RD. A polymethyl methacrylate method for large specimens of mineralized bone with implants.
  • a Kaplan-Meier survivorship curve was used to compare infection rates in each rabbit group.
  • the rabbit was classified as infected if it had at least two of the three infection criteria: 1) greater than Grade I clinical signs of infection, 2) positive culture for one or more of the tissue samples, and/or 3) positive histology indicative of infection for one or more of the tissue samples.
  • Statistical differences between the groups were tested with a log-rank test for equality of survivor functions. A p-value of ⁇ 0.05 was considered statistically significant.
  • the second limitation was the high drop out rate of the porous tantalum implants in Group 3 due to breaking and/or having the skin heal over the implant resulting.
  • the low strength of porous tantalum in bending may have contributed to its breaking.
  • Porous tantalum's ultimate bending strength has been reported as 110 ⁇ 14 MPa for a disk 2.5 x
  • pexiganan acetate has been shown to kill gram-positive and gram- negative anaerobic and aerobic bacteria.
  • pexiganan incorporated into collagen matrices reduced P. aeruginosa and S. aureus growth in infected wounds on rats.
  • the current study supported pexiganan acetate's effectiveness against gram-positive and gram-negative bacteria since S. aureus and E.
  • pexiganan acetate is an important antimicrobial for transcutaneous osseointegrated implants since it helped to prevent organisms from traversing the skin barrier into muscle, blood, and bone.
  • Porous tantalum, as a coating on an osseointegrated implant, will not likely prevent pin track infection without additional methods of soft tissue immobilitization around the implant site

Landscapes

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

Abstract

La présente invention concerne un procédé de prévention de l'infection d'une plaie par administration d'une forme galénique thérapeutique, comprenant un peptide, sur le site d'une plaie. Plus précisément, l'invention concerne l'utilisation de peptides amphiphiles dans le cadre de la prévention et du traitement d'infections. L'invention concerne également la prévention de l'infection d'une plaie chez un hôte grâce à l'administration d'un peptide amphiphile biologiquement actif ou d'une protéine biologiquement active chez un hôte porteur d'une plaie, le peptide ou la protéine étant administré en quantité efficace pour prévenir l'infection de la plaie. Plus précisément encore, l'invention concerne l'utilisation de peptides amphiphiles cationiques dans le cadre de la prévention et du traitement d'infections associées à des implants transcutanés ostéointégrés.
PCT/US2008/084640 2007-11-27 2008-11-25 Compositions et procédés utilisables dans le cadre de la prévention d'infections associées à des implants transcutanés ostéointégrés WO2009070564A2 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US99043007P 2007-11-27 2007-11-27
US60/990,430 2007-11-27

Publications (2)

Publication Number Publication Date
WO2009070564A2 true WO2009070564A2 (fr) 2009-06-04
WO2009070564A3 WO2009070564A3 (fr) 2009-08-27

Family

ID=40679204

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2008/084640 WO2009070564A2 (fr) 2007-11-27 2008-11-25 Compositions et procédés utilisables dans le cadre de la prévention d'infections associées à des implants transcutanés ostéointégrés

Country Status (1)

Country Link
WO (1) WO2009070564A2 (fr)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2012174373A1 (fr) * 2011-06-16 2012-12-20 Dipexium Pharmaceuticals Méthodes de traitement et d'inhibition d'agents pathogènes avec des mécanismes de résistance
CN104602762A (zh) * 2012-06-12 2015-05-06 帝普休姆制药有限公司 稳定的培西加南配方
WO2020043834A1 (fr) * 2018-08-30 2020-03-05 Stichting Katholieke Universiteit Feuille de polyuréthane
NL2021630B1 (en) * 2018-09-14 2020-05-06 Stichting Katholieke Univ Polyurethane sheet

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5045531A (en) * 1989-12-18 1991-09-03 Magainin Sciences Inc. Wound treatment employing biologically active ion channel forming peptides and proteins
US5792831A (en) * 1990-02-08 1998-08-11 Magainin Pharmaceuticals Inc. Analogues of magainin peptides containing D-amino acids

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2012174373A1 (fr) * 2011-06-16 2012-12-20 Dipexium Pharmaceuticals Méthodes de traitement et d'inhibition d'agents pathogènes avec des mécanismes de résistance
CN104602762A (zh) * 2012-06-12 2015-05-06 帝普休姆制药有限公司 稳定的培西加南配方
JP2015521599A (ja) * 2012-06-12 2015-07-30 ディペキシウム ファーマシューティカルズ インコーポレイテッド 安定なペキシガナン製剤
EP2858721A4 (fr) * 2012-06-12 2015-12-09 Dipexium Pharmaceuticals Inc Formulation stable de pexiganan
WO2020043834A1 (fr) * 2018-08-30 2020-03-05 Stichting Katholieke Universiteit Feuille de polyuréthane
NL2021630B1 (en) * 2018-09-14 2020-05-06 Stichting Katholieke Univ Polyurethane sheet

Also Published As

Publication number Publication date
WO2009070564A3 (fr) 2009-08-27

Similar Documents

Publication Publication Date Title
Diefenbeck et al. Gentamicin coating of plasma chemical oxidized titanium alloy prevents implant-related osteomyelitis in rats
Gao et al. Enhancing antibacterial capability and osseointegration of polyetheretherketone (PEEK) implants by dual-functional surface modification
ES2952858T3 (es) Nuevos polipéptidos y uso médico de los mismos
RU2695455C2 (ru) Противомикробный пептид
EP3280722B1 (fr) Peptides antimicrobiens et leur utilisation pour le traitement d'infections locales
Chou et al. Evaluating antimicrobials and implant materials for infection prevention around transcutaneous osseointegrated implants in a rabbit model
Rammelt et al. In vivo effects of coating loaded and unloaded Ti implants with collagen, chondroitin sulfate, and hydroxyapatite in the sheep tibia
WO2012023510A1 (fr) Implant biologique
Schindeler et al. Local delivery of the cationic steroid antibiotic CSA-90 enables osseous union in a rat open fracture model of Staphylococcus aureus infection
Perry et al. Assessing peri‐implant tissue infection prevention in a percutaneous model
WO2005006938A2 (fr) Implants medicaux antimicrobiens et utilisations associees
JP2007332040A (ja) カーボンナノチューブを含む抗菌剤とそれを用いた材料及び製剤
WO2009070564A2 (fr) Compositions et procédés utilisables dans le cadre de la prévention d'infections associées à des implants transcutanés ostéointégrés
Peng et al. Quaternised chitosan coating on titanium provides a self-protective surface that prevents bacterial colonisation and implant-associated infections
KR101821842B1 (ko) Ｐｇａ를 포함하는 새로운 ｅｍｄ 제제
KR101493437B1 (ko) 컨센서스 펩티드
CN105963680A (zh) 用于抑制/瓦解生物被膜的抑制剂及其应用
Stallmann et al. In vivo release of the antimicrobial peptide hLF1‐11 from calcium phosphate cement
EP4228677A1 (fr) Nouvelles combinaisons peptidiques bioactives et leurs utilisations
Lin et al. Augmentation of osseous phenotypes in vivo with a synthetic peptide
WO2018115877A2 (fr) Compositions et méthodes de traitement
Kazemzadeh-Narbat et al. Antimicrobial peptide delivery from trabecular bone grafts
US20220296669A1 (en) Novel compositions and uses thereof
US20250134946A1 (en) Self-assembling amphiphilic peptide hydrogels for treatment of nerve injury
Zeng et al. Titanium oxide nanotubes embedded with silver dioxide nanoparticles for Staphylococcus aureus infections after prosthetic joint replacement in animal models

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 08855099

Country of ref document: EP

Kind code of ref document: A2

NENP Non-entry into the national phase

Ref country code: DE

32PN Ep: public notification in the ep bulletin as address of the adressee cannot be established

Free format text: NOTING OF LOSS OF RIGHTS PURSUANT TO RULE 112(1) EPC (EPO FORM 1205A DATED 22.10.2010)

122 Ep: pct application non-entry in european phase

Ref document number: 08855099

Country of ref document: EP

Kind code of ref document: A2

点击 这是indexloc提供的php浏览器服务，不要输入任何密码和下载