WO2006076254A2 - Methode de traitement ou de prevention d'une infection microbienne du tissu respiratoire - Google Patents

Methode de traitement ou de prevention d'une infection microbienne du tissu respiratoire Download PDF

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Publication number
WO2006076254A2
WO2006076254A2 PCT/US2006/000592 US2006000592W WO2006076254A2 WO 2006076254 A2 WO2006076254 A2 WO 2006076254A2 US 2006000592 W US2006000592 W US 2006000592W WO 2006076254 A2 WO2006076254 A2 WO 2006076254A2
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WO
WIPO (PCT)
Prior art keywords
tissue
lkktet
respiratory
peptide
infection
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Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/US2006/000592
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English (en)
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WO2006076254A3 (fr
Inventor
Allan L. Goldstein
Jack Finkelstein, Jr.
David Crockford
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RegeneRx Biopharmaceuticals Inc
Original Assignee
RegeneRx Biopharmaceuticals 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 RegeneRx Biopharmaceuticals Inc filed Critical RegeneRx Biopharmaceuticals Inc
Publication of WO2006076254A2 publication Critical patent/WO2006076254A2/fr
Priority to US11/773,168 priority Critical patent/US20080051348A1/en
Anticipated expiration legal-status Critical
Publication of WO2006076254A3 publication Critical patent/WO2006076254A3/fr
Ceased legal-status Critical Current

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
    • A61K38/08—Peptides having 5 to 11 amino acids

Definitions

  • the present invention relates to the field of treating or preventing respiratory microbial infection of respiratory tissue.
  • Respiratory infections caused by Pseudomonas aeruginosa and other gram- negative bacteria occur almost exclusively in individuals with a compromised lower respiratory tract or a compromised systemic defense mechanism.
  • Primary pneumonia occurs in patients with chronic lung disease and congestive heart failure.
  • Bacteremic pneumonia commonly occurs in neutropenic cancer patients undergoing chemotherapy.
  • Lower respiratory tract colonization of cystic fibrosis patients by mucoid strains of Pseudomonas aeruginosa and other gram-negative bacteria is common and difficult, if not impossible, to treat.
  • a method of treatment for treating, preventing, inhibiting or reducing respiratory microbial infection of respiratory tissue of a subject comprises administering to a subject in need of such treatment an effective amount of a composition comprising an antimicrobial agent comprising amino acid sequence LKKTET or LKKTNT, a conservative variant thereof, or a stimulating agent that stimulates production of an LKKTET or LKKTNT peptide, or a conservative variant thereof, in said tissue, so as to inhibit said microbial infection.
  • actin-sequestering peptides such as thymosin, beta 4 (T ⁇ 4 or TB4) and other agents including actin-sequestering peptides or peptide fragments containing amino acid sequence LKKTET or LKKTNT or conservative variants thereof, promote reversal or prevention of respiratory infection of respiratory tissue.
  • a respiratory infection treated in accordance with the present invention is a bacterial infection, more preferably a gram-negative bacterial infection, and most preferably a Pseudomonas aeruginosa respiratory infection.
  • a subject being treated in accordance with the present invention preferably is mammalian, most preferably human.
  • Thymosin 4 was initially identified as a protein that is up-regulated during endothelial cell migration and differentiation in vitro. Thymosin 4 was originally isolated from the thymus and is a 43 amino acid, 4.9 kDa ubiquitous polypeptide identified in a variety of tissues. Several roles have been ascribed to this protein including a role in a endothelial cell differentiation and migration, T cell differentiation, actin sequestration, vascularization and wound healing.
  • the invention is a method of treatment for treating, preventing, inhibiting or reducing respiratory microbial infection of respiratory tissue of a subject, comprising administering to a subject in need of such treatment an effective amount of a composition comprising an antimicrobial agent, which may be a polypeptide comprising amino acid sequence LKKTET or LKKTNT, or a conservative variant thereof having antimicrobial activity, preferably Thymosin ⁇ 4, and/or T ⁇ 4 isoforms, analogues or derivatives, including KLKKTET, LKKTETQ, N-terminal variants of T ⁇ 4, C-terminal variants of T ⁇ 4 and antagonists of T ⁇ 4.
  • the invention also may utilize oxidized T ⁇ 4.
  • the antimicrobial agent is other than thymosin beta 4 or oxidized T ⁇ 4.
  • T ⁇ 4 Thymosin ⁇ 4
  • T ⁇ 4 Thymosin ⁇ 4
  • T ⁇ 4 Thymosin ⁇ 4
  • PCT/US99/17282 discloses isoforms of T ⁇ 4 which may be useful in accor ⁇ ance witn the present invention as well as amino acid sequence LKKTET and conservative variants thereof, which may be utilized with the present invention.
  • International Application Serial No. PCT/GB99/00833 discloses oxidized Thymosin ⁇ 4 which may be utilized in accordance with the present invention.
  • T ⁇ 4 and T ⁇ 4 isoforms are described primarily hereinafter with respect to T ⁇ 4 and T ⁇ 4 isoforms, it is to be understood that the following description is intended to be equally applicable to amino acid sequence LKKTET or LKKTNT, peptides and fragments comprising or consisting essentially of LKKTET or LKKTNT, conservative variants thereof having antimicrobial activity, and/or T ⁇ 4 isoforms, analogues or derivatives, including N-terminal variants of T ⁇ 4, C-terminal variants of T ⁇ 4 and antagonists of T ⁇ 4.
  • the invention also may utilize oxidized T ⁇ 4.
  • the antimicrobial agent may be directly or indirectly antimicrobial.
  • the invention provides a method of treatment for treating, preventing, inhibiting or reducing respiratory microbial infection of respiratory tissue of a subject, by contacting the tissue with an antimicrobial effective amount of a composition which contains an antimicrobial agent as described herein.
  • the tissue may be selected from respiratory tract or airway tissue of said subject.
  • the contacting may be directly or systemlcally.
  • direct administration include, for example, contacting the tissue, by direct application or inhalation, with a solution, lotion, salve, gel, cream, paste, spray, suspension, dispersion, hydrogel, ointment, or oil comprising an antimicrobial agent as described herein.
  • Systemic administration includes, for example, intravenous, intraperitoneal, intramuscular injections or infusions of a composition containing an antimicrobial agent as described herein, in a pharmaceutically acceptable carrier such as water for injection.
  • Antimicrobial agents for use in the invention, as described herein may be administered in any suitable microbial infection-inhibiting amount.
  • an antimicrobial agent as described herein may be administered in dosages within the range of about 0.0001-1 ,000,000 micrograms, more preferably in amounts within the range of about 0.1-5,000 micrograms, most preferably within the range of about 1-30 micrograms.
  • a composition in accordance with the present invention can be administered daily, every other day, every other week, every other month, etc., with a single application or multiple applications per day of administration, such as applications 2, 3, 4 or more times per day of administration.
  • Many T ⁇ 4 isoforms have been identified and have about 70%, or about 75%, or about 80% or more homology to the known amino acid sequence of T ⁇ 4.
  • Such isoforms include, for example, T ⁇ 4 ala , T ⁇ 9, T ⁇ 10, T ⁇ 11 , T ⁇ 12, T ⁇ 13, T ⁇ 14 and T ⁇ 15. Similar toT ⁇ 4, the T ⁇ 10 and T ⁇ 15 isoforms have been shown to sequester actin.
  • T ⁇ 4, T ⁇ 10 and T ⁇ 15, as well as these other isoforms share an amino acid sequence, LKKTET or LKKTNT, that appears to be involved in mediating actin sequestration or binding.
  • T ⁇ 4 can modulate actin polymerization (e.g. ⁇ -thymosins appear to depolymerize F-actin by sequestering free G-actin).
  • T ⁇ 4's ability to modulate actin polymerization may therefore be due to all, or in part, its ability to bind to or sequester actin via the LKKTET sequence.
  • T ⁇ 4 other proteins which bind or sequester actin, or modulate actin polymerization, including T ⁇ 4 isoforms having the amino acid sequence LKKTET, are likely to be effective, alone or in a combination with T ⁇ 4, as set forth herein.
  • T ⁇ 4 isoforms such as T ⁇ 4 ala , T ⁇ 9, T ⁇ 10, T ⁇ 11 , T ⁇ 12, T ⁇ 13, T ⁇ 14 and T ⁇ 15, as well as T ⁇ 4 isoforms not yet identified, will be useful in the methods of the invention.
  • T ⁇ 4 isoforms are useful in the methods of the invention, including the methods practiced in a subject.
  • the invention therefore further provides pharmaceutical compositions comprising T ⁇ 4, as well as T ⁇ 4 isoforms T ⁇ 4 ala , T ⁇ 9, T ⁇ 10, T ⁇ 11 , T ⁇ 12, T ⁇ 13, T ⁇ 14 and T ⁇ 15, and a pharmaceutically acceptable carrier.
  • antimicrobial agents or proteins having actin sequestering or binding capability or that can mobilize actin or modulate actin polymerization, as demonstrated in an appropriate sequestering, binding, mobilization or polymerization assay, or identified by the presence of an amino acid sequence that mediates actin binding, such as LKKTET or LKKTNT, for example, can similarly be employed in the methods of the invention.
  • Such proteins may include gelsolin, vitamin D binding protein (DBP), profilin, cofilin, depactin, Dnasel, vilin, fragmin, severin, capping protein,, ⁇ - actinin and acumentin, for example.
  • the invention further provides pharmaceutical compositions comprising gelsolin, vitamin D binding protein (DBP), profilin, cofilin, depactin, Dnasel, vilin, fragmin, severin, capping protein, ⁇ -actinin and acumentin as set forth herein.
  • DBP vitamin D binding protein
  • the invention includes the use of an antimicrobial polypeptide comprising the amino acid sequence LKKTET or LKKTNT and conservative variants thereof.
  • conservative variants thereof denotes the replacement of an amino acid residue by another, biologically similar residue.
  • T ⁇ 4 has been localized to a number of tissue and cell types and thus, agents which stimulate the production of an LKKTET or LKKTNT peptide such as T ⁇ 4 or another antimicrobial agent as described herein, can be added to or comprise a composition to effect production an antimicrobial agent from a tissue and/or a cell.
  • agents which stimulate the production of an LKKTET or LKKTNT peptide such as T ⁇ 4 or another antimicrobial agent as described herein, can be added to or comprise a composition to effect production an antimicrobial agent from a tissue and/or a cell.
  • Such stimulating agents may include members of the family of growth factors, such as insulin-like growth factor (IGF-1), platelet derived growth factor (PDGF), epidermal growth factor (EGF), transforming growth factor beta (TGF- ⁇ ), basic fibroblast growth factor (bFGF), thymosin cc1 (TaI) and vascular endothelial growth factor (VEGF). More preferably, the stimulating agent is transforming growth factor beta (TGF.- ⁇ ) or other members of the TGF.- ⁇ superfamily.
  • IGF-1 insulin-like growth factor
  • PDGF platelet derived growth factor
  • EGF epidermal growth factor
  • TGF- ⁇ transforming growth factor beta
  • bFGF basic fibroblast growth factor
  • TaI thymosin cc1
  • VEGF vascular endothelial growth factor
  • subjects are treated with a stimulating agent that stimulates production in the subject of an antimicrobial agent as defined herein.
  • an antimicrobial agent as described herein alone or in combination can be added in combination with any one or more of the following agents: antibiotics, VEGF, KGF, FGF, PDGF, TGF ⁇ , IGF-1 , IGF- 2, IL-I , prothymosin ⁇ and/or thymosin ⁇ 1 in an effective amount.
  • the invention also includes a pharmaceutical composition comprising a therapeutically effective amount of an antimicrobial agent as described herein in a pharmaceutically acceptable carrier such as water for injection.
  • Suitable formulations may include an antimicrobial agent as described herein at a concentration within the range of about 0.001 - 50% by weight, more preferably within the range of about 0.01 - 0.1% by weight, most preferably about 0.05% by weight.
  • the therapeutic approaches described herein involve various routes of administration or delivery of an antimicrobial agent as described herein, including any conventional administration techniques (for example, but not limited to, direct administration, local injection, inhalation, or systemic administration), to a subject.
  • the methods and compositions using or containing an antimicrobial agent as described herein may be formulated into pharmaceutical compositions by admixture with pharmaceutically acceptable non-toxic excipients or carriers.
  • the invention may include use of antibodies which interact with an antimicrobial agent as described herein. Antibodies which consist essentially of pooled monoclonal antibodies with different epitopic specificities, as well as distinct monoclonal antibody preparations are provided.
  • Monoclonal antibodies are made from antigen containing fragments of the protein by methods well known to those skilled in the art as disclosed in PCT/US99/17282, supra.
  • the term antibody as used in this invention is meant to include monoclonal and polyclonal antibodies.
  • the invention provides a method of treating a subject by administering an effective amount of stimulating agent which modulates gene expression.
  • modulate refers to inhibition or suppression of expression when an antimicrobial agent as described herein is over expressed, and induction of expression when an antimicrobial agent as described herein is underexpressed.
  • effective amount means that amount of stimulating agent which is effective in modulating gene expression of an antimicrobial agent as described herein, resulting in reducing the symptoms of respiratory microbial infection of respiratory tissue.
  • a stimulating agent which modulates gene expression of a response-inhibiting agent as described herein may be a polynucleotide, for example.
  • the polynucleotide may be an antisense, a triplex agent, or a ribozyme.
  • an antisense directed to the structural gene region or to the promoter region of an antimicrobial agent as described herein may be utilized.
  • the stimulating agent which modulates gene expression of an antimicrobial agent as described herein may also be a small interfering RNAs (siRNAs).
  • siRNAs small interfering RNAs
  • the invention provides a method for utilizing compounds that modulate activity of an antimicrobial agent as described herein.
  • Compounds that affect activity of an antimicrobial agent as described herein include peptides, peptidomimetics, polypeptides, chemical compounds, minerals such as zincs, and biological agents.
  • a method for screening for a stimulating agent as defined herein comprises contacting a respiratory tissue exhibiting respiratory microbial infection, with a candidate compound; and measuring activity in said tissue of an LKKTET or LKKTNT peptide, wherein an increase of activity of said peptide in said tissue, compared to a level of activity of said peptide in a corresponding tissue lacking said candidate compound, indicates that said compound is capable of inducing said stimulating agent.
  • a further method of screening for a stimulating agent as defined herein comprises contacting a respiratory tissue with a candidate compound, optionally microbially infecting the tissue, and measuring LKKTET or LKKTNT peptide activity in said tissue, wherein an increase of activity in said tissue, compared to a level of said LKKTET or LKKTNT peptide activity in a corresponding tissue lacking said candidate compound, indicates that said candidate compound is capable of stimulating production in said tissue of said peptide.
  • Antimicrobial assays were performed to assess peptide activity against Pseudomonas aeruginosa (PA), Staphylococcus aureus (PA), and Staphylococcus epidermidis (SE).
  • PA Pseudomonas aeruginosa
  • PA Staphylococcus aureus
  • SE Staphylococcus epidermidis

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

Abstract

L'invention concerne une méthode destinée au traitement, à la prévention et à l'inhibition ou à l'atténuation d'une infection microbienne du tissu respiratoire chez un patient, consistant à administrer à un patient nécessitant un tel traitement une dose utile d'une composition contenant un agent antimicrobien renfermant une séquence d'acide aminé LKKTET ou LKKTNT, une variante conservatrice de cette séquence, ou un agent stimulant la production d'un peptide LKKTET ou LKKTNT, ou une variante conservatrice de ce peptide, dans le tissu, de manière à inhiber l'infection.
PCT/US2006/000592 2002-02-06 2006-01-10 Methode de traitement ou de prevention d'une infection microbienne du tissu respiratoire Ceased WO2006076254A2 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
US11/773,168 US20080051348A1 (en) 2002-02-06 2007-07-03 Treatment of infections and other disorders

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US64252005P 2005-01-11 2005-01-11
US60/642,520 2005-01-11

Related Child Applications (1)

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US11/773,168 Continuation-In-Part US20080051348A1 (en) 2002-02-06 2007-07-03 Treatment of infections and other disorders

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WO2006076254A2 true WO2006076254A2 (fr) 2006-07-20
WO2006076254A3 WO2006076254A3 (fr) 2007-10-25

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Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003020215A2 (fr) * 2001-08-29 2003-03-13 Regenerx Biopharmaceuticals, Inc. Procedes de guerison ou de prevention de l'inflammation, de la deterioration ou d'autres modifications intervenant avant, pendant ou immediatement apres un evenement myocardique par thymosine beta 4, ses analogues, isoformes ou autres derives
AU2003296875B2 (en) * 2002-02-06 2008-01-24 Regenerx Biopharmaceuticals, Inc. Treatment of infections and other disorders

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