WO2001034641A2 - Antimicrobial activity of the first cationic cluster of human lactoferrin - Google Patents
Antimicrobial activity of the first cationic cluster of human lactoferrin Download PDFInfo
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- WO2001034641A2 WO2001034641A2 PCT/NL2000/000821 NL0000821W WO0134641A2 WO 2001034641 A2 WO2001034641 A2 WO 2001034641A2 NL 0000821 W NL0000821 W NL 0000821W WO 0134641 A2 WO0134641 A2 WO 0134641A2
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- polypeptide
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/06—Lysis of microorganisms
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/79—Transferrins, e.g. lactoferrins, ovotransferrins
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Definitions
- the present invention relates to the field of polypeptides having various therapeutic and prophylactic applications, including bactericidal activity. As such, the present invention broadly relates to the fields of protein and medicinal chemistry.
- Lactoferrin is a metal binding glycoprotein of Mr 77,000 found in milk, tears, saliva, bronchial, intestinal, and other secretions. LF is also present in the secondan granules of neutrophils. Lactoferrin plays an important role in numerous inflammatory and immune response functions such as regulation of monocyte colony stimulating factor synthesis, activation of natural killer cell activity, inhibition of metastasis, and maturation of T-cells. Lactoferrin also inhibits myelopoiesis, binds to members of the low density lipoprotein receptor family, and blocks the clearance of lipoprotein chylomicron remnant particles (2, 32, 33, 34).
- hLF Human lactoferrin
- Human LF consists of a polypeptide chain of 692 amino acids and contains two N-terminal cationic domains, i.e., RRRR (residues 2-5 of SEQ ID NO:l) and RKVR (residues 28-31 of SEQ ID NO:l), whereas bovine lactoferrin (bLF) has only one cationic domain (residues 17-42 (8,9)).
- the LF polypeptide is folded into two globular lobes, each of which contains an iron-binding cleft.
- the high affinity of LF for iron confers to the protein certain antibacterial properties and, in addition, may play a role in the absorption of dietary iron by the small intestine.
- This invention provides polypeptides and pharmaceutical compositions that are useful in treating a wide variety of microbial infections such as bacterial infections and therapeutic methods for using such compositions.
- the present invention provides polypeptides that include at least 6 but no more than 27 contiguous amino acids from the N-terminal segment of human lactoferrin protein (SEQ ID NO: l), wherein the N-terminus of said polypeptide is residue 1 of SEQ ID NO: 1.
- the invention further includes similar polypeptides that include at least 6 but not more than 26 or 25 contiguous amino acids from the N-terminal segment of human lactoferrin protein, wherein the N-terminus of said polypeptide is residue 2 and 3 of SEQ ID NO: 1 , respectively.
- Specific examples of polypeptides provided by the invention include hLF(l-l l ), hLF(2-l 1) and hLF(3-l 1).
- the present invention further provides a variety of pharmaceutical compositions that in general include a polypeptide as described below and a pharmaceutically acceptable excipient.
- Certain polypeptides used in some pharmaceutical compositions are no more than 27 amino acids in length, have an N-terminal amino acid sequence of XaaArgArg, (Xaa is any amino acid and Arg is the amino acid arginine) and have antimicrobial activity.
- Other polypeptides used in other pharmaceutical compositions of the invention are similar to that just described, except they are no more than 26 amino acid residues in length and have an N-terminal sequence of ArgArg.
- Still other related polypeptides are 25 amino acids or less in length and have an N-terminal Arg.
- Polypeptides that can be included in other pharmaceutical compositions include any of the polypeptides described herein, including, for example, various N-terminal fragments from hLF (e.g., hLF(l-l 1), hLF(2-l 1) and hLF(3- 1 1)).
- the invention also provides methods for treating patients suffering from various microbial infections, such as bacterial, viral, fungal or parasitic infections, for example.
- the methods include administering a therapeutic amount or prophylatically effective amount of a polypeptide or pharmaceutical composition of the invention to a patient.
- Methods for detecting microbial infections in a patient are also provided by the invention. These methods typicalh involve administering a labeled polypeptide to a patient infected with, or suspected of being infected with, a microbe, wherein said polypeptide is human lactoferrin protein (SEQ ID NO: l ) or a subsequence thereof capable of interacting with said microbe and detecting the presence of said labeled polypeptide at a site of infection.
- the polypeptide is labeled with a radioisotope and the presence of the radioisotope at the site of infection is visualized with a gamma camera.
- the invention further provides methods for altering the permeability of microbial cell membrane by contacting the microbial cell membrane with a polypeptide of the invention.
- FIGS. 1A and I B are charts depicting the antibacterial activity of natural hLF and hLF ⁇ 3N (hLF protein without the N-terminal glycine and the two N-terminal arginines, i.e., without residues 1-3 of SEQ ID NO: l) against L. monocytogenes, and antibiotic resistant S. aureiis.
- FIGS. 1 A and IB illustrate a dose-dependent reduction of the number of viable __. m ⁇ nocylogenes (FIG. 1A), and antibiotic resistant S. aureus (FIG. IB) by natural hLF and hLF "3N .
- Tests involved exposing approximately 1 -2x10° CFU of __. monocytogenes or antibiotic resistant S. aureus to various concentrations of natural hLF or hLF " for 3 h at 37 °C; the number of viable bacteria remaining at that point were then determined using standard microbiological techniques. The detection limit was 4,000 CFU. Results are expressed as a mean value ( ⁇ standard deviation (SD)) and are calculated from at least three separate experiments.
- FIGS. 1C-1E show a time-dependent reduction of the number of viable __. monocytogenes (FIG. 1C), antibiotic resistant S. aureus (FIG. ID) and K. pneumoniae
- FIGS. 2A-2D are graphs showing the relative abilities of hLF(l-l 1 ) (residues 1-1 1 inclusively from the N-terminus of hLF, i.e., residues 1-1 1 of SEQ ID NO: 1) and fragments thereof, as well as hLF(21-31) (residues 21 -31 from the N-terminal region of hLF, i.e., residues 21-31 of SEQ ID NO: l), to kill ofZ. monocytogenes (FIG. 2A), antibiotic resistant S. aureus (FIG 2B), A ' , pneumoniae (FIG. 2C) and E. coli (FIG. 2D).
- FIG. 3A-3C illustrate the effect of hLF(l -l 1 ) and fragments thereof and hLF(21-31) on L. monocytogenes (FIG. 3A), antibiotic resistant S. aureus (FIG. 3B), and E. coli (FIG. 3C).
- Approximately lxl 0 6 CFU of the various bacteria were exposed to increasing micromolar concentrations of these peptides for 1 h at 37 °C. The number of viable bacteria was determined microbiologically and the percentage of propidium iodide-positive bacteria was determined by FACS analysis. Results are expressed as a mean value ( ⁇ SD) and are calculated from at least three separate experiments.
- FIG. 4 depicts the effect of natural hLF and hLF "3N on the non-specific esterase activity of __. monocytogenes.
- Approximately 2xl0 6 CFU of bacteria were initially incubated with 4 ⁇ M of natural hLF, hLF "3N , or BSA (-) for 2 h at 37 °C, and then incubated with 100 ⁇ M of 5-sulfofluorescein diacetate in 20% (v/v) ethanol for 20 min at room temperature in the dark. The bacteria were then washed and fluorescence intensity assessed by FACS analysis.
- FIG. 5 shows the bactericidal activity of natural hLF, hLF "3N and various related peptides in mice infected with antibiotic resistant S. aureus .
- Mice were intramuscularly infected with approximately lxlO 6 CFU of antibiotic resistant S. aureus in 0.1 ml of saline. Twenty-four hours thereafter, 0.2 ml of saline containing various amounts (0.1-10 n ole) of natural hLF, hLF "3N , hLF(l-l 1), hLF(4-l l) or hLF(21-31) was injected intravenously.
- FIG. 6 shows accumulation of 99m Tc-hLF or related peptides in mice experimentally infected with antibiotic resistant S. aureus.
- Results are expressed as a mean value ( ⁇ SD) of at least three mice (p ⁇ 0.05), compared with m Tc-lgG .
- polypeptide peptide
- protein protein
- amino acid polymers in which one or more amino acids are chemical analogues of a corresponding naturally-occurring amino acid.
- human lactoferrin protein refers to a full-length human lactoferrin polypeptide, e.g., a polypeptide having an amino acid sequence substantially as described by Powell, M.J. and Ogden, J.E., Nucleic Acids Res. 18:4013 (1990), which is incorporated herein by reference.
- An exemplary sequence for human lactoferrin protein is the sequence set forth in SEQ ID NO: 1.
- Other related human lactoferrin sequences are provided, for example, by Metz-Boutigue, M.H., et al., Eur. J. Biochem.
- human lactoferrin protein also includes naturally-occurring human allelic variants and variants involving conservative substitutions of amino acids.
- human lactoferrin protein also includes hLF wherein the protein backbone is modified. Examples of such modifications include acetylations, carboxylation, glycosylation modifications and other processing variants of hLF.
- natural human lactoferrin includes recombinantly encoded human lactoferrin expressed in a transgenic nonhuman animal, such as a bovine, where the glycosylation pattern may be distinct from glycosylation patterns of naturally-occurring human lactoferrin obtained from human milk.
- hLF fragment refers to a polypeptide wherein the amino acid residues of the polypeptide consist of a contiguous sequence of amino acids from hLF.
- these terms include polypeptides having conservatively modified variations or chemical modifications (e.g., carboxylation, glycosylation, acetylations) to a subsequence of hLF.
- the term also includes fragments wherein the cysteine at residue 10 (see SEQ ID NO: l ) is conserved, in both fragments consisting of contiguous sequence of amino acids from hLF as well as fragments having conservative variations.
- hLF( 1 -1 1 ) refers to residues 1 to 1 1 inclusively from the N-terminus of hLF; similarly, hLF(2-l 1) refers to residues 2 to 1 1 inclusively from the N-terminal region of hLF.
- hLF i.e., the full-length protein
- hLF in which the N-terminal glycine and arginine are removed is referred to as hLF " N ; hLF missing the N- terminal glycine and the two N-terminal arginines, is referred to as hLF ' , and hLF lacking the N-terminal glycine and first three arginine residues is referred to as hLF " N .
- the three arginines located at the N-terminus i.e., residues 2, 3 and 4 of SEQ ID NO: l
- Arg residues 2, 3 and 4 of SEQ ID NO: l
- N-terminal amino acid of hLF refers to Gly' (see SEQ ID NO:l); the 31 residues located at the amino-terminus of hLF are: N'-GRRRRSVQWCAVSQPEATKCFQWQRNMRKVR (residues 1 -31 of SEQ ID NO: l ).
- “conservatively modified variations” of a particular amino acid sequence refers to amino acid substitutions of those amino acids that are not critical for protein activity or substitution of amino acids with other amino acids having similar properties (e.g., acidic, basic, positively or negatively charged, polar or non-polar) such that the substitutions of even critical amino acids do not substantially alter activity.
- Conservative substitution tables providing functionally similar amino acids are well-known in the art. See, e.g., Creighton (1984) Proteins, W.H. Freeman and Company.
- individual substitutions, deletions or additions which alter, add or delete a single amino acid or a small percentage of amino acids in an encoded sequence are also “conservatively modified variations.”
- naturally-occurring refers to the fact that an object can be found in nature.
- polypeptide sequence that is present in an organism that can be isolated from a source in nature and which has not been intentionally modified by humans in the laboratory is naturally-occurring.
- isolated means an object species (e.g. , hLF or a fragment thereof) is the predominant macromolecular species present (i.e., on a molar basis it is more abundant than any other individual species in the composition), and preferably the object species comprises at least about 50 percent (on a molar basis) of all macromolecular species present.
- the object species in an isolated, purified or substantially pure composition will comprise more than 80 to 90 percent of all macromolecular species present in a composition.
- the object species is purified to essential homogeneity (i.e., contaminant species cannot be detected in the composition by conventional detection methods) wherein the composition consists essentially of a single macromolecular species.
- a specified antibody binds preferentially to a particular protein and does not bind in a significant amount to other proteins present in the sample.
- Specific binding to a protein under such conditions requires an antibody that is selected for its specificity for a particular protein.
- a molecule such as an antibody that specifically binds to a protein has an association constant of at least 10° M " 1 or 10 7 M '1 , preferably 10 8 M "1 to 10 9 M "1 .
- immunoassay formats may be used to select antibodies specifically immunoreactive with a particular protein.
- solid-phase ELISA immunoassays are routinely used to select monoclonal antibodies specifically immunoreactive with a protein. See, e.g., Harlow and Lane (1988) Antibodies, A Laboratory Manual, Cold Spring Harbor Publications, New York, for a description of immunoassay formats and conditions that can be used to determine specific immunoreactivity.
- patient includes human and veterinary subjects.
- the invention provides polypeptides such as hLF fragments and pharmaceutical compositions utilizing such polypeptides that are useful in a variety of therapeutic and prophylactic applications, including use as antimicrobial agents.
- the invention is based in part upon the finding that short polypeptides having one or more arginines in the N-terminal segment of the polypeptide, such as found in the first cationic domain of hLF. exhibit significant therapeutic activity.
- polypeptides of the invention include one or more residues from the first cationic domain of hLF, but do not include amino acids from the second cationic domain
- the polypeptides are quite short, generally less than 27 amino acids in length. Given their short length, the polypeptides are easily and inexpensively prepared and are readily amenable to use in pharmaceutical compositions. Antibodies which specifically bind to the polypeptides of the invention are also provided.
- the invention further provides pharmaceutical compositions which include a polypeptide and one or more other components such as a pharmaceutically acceptable excipient.
- the polypeptide used in the composition is a short polypeptide typically less than 27 amino acids long, has one or more arginine residues in the N-terminal segment, and has antimicrobial activity.
- the polypeptide can consist of contiguous sequences from the first cationic domain of hLF. for example.
- Methods of treating patients suffering from infections are further provided by the invention.
- the methods involve administering a therapeutic dose of one of the polypeptides or pharmaceutical compositions of the invention to a patient.
- the methods are effective against a wide range of microbes, including, for example, viruses and bacteria.
- the invention also discloses methods for detecting or imaging microbial infections such as bacterial infections using labeled hLF or fragments thereof which migrate to the site of a microbial infection. Using a detector suitable for the type of label attached to the peptide, it is possible to detect infection sites.
- the invention provides various polypeptides which include at least one or more arginines from the first cationic domain of hLF (residues 2-5 of SEQ ID NO: l), but which exclude the residues which make up the second cationic domain (residues 28-31 of SEQ ID NO:l).
- the invention provides polypeptides which comprise at least 6 but no more than 27 contiguous amino acids from the N-terminal segment of hLF (SEQ ID NO: l), wherein the N-terminus of the polypeptide is residue 1 of SEQ ID NO: l, i.e., glycine.
- the invention further includes similar polypeptides that include the first cationic domain but not the second cationic domain of hLF, and that also lack one or more residues from the N-terminus of hLF.
- the invention also includes polypeptides comprising at least 6 amino acids but no more than 26 contiguous amino acids from hLF, wherein the N-terminus of the polypeptide is Arg " (for example, a hLF fragment lacking Gly 1 ).
- the invention also includes polypeptides comprising at least 6 but no more than 25 contiguous amino acids from the N-terminal segment of hLF. wherein the N-terminus of the polypeptide is Arg ' ' (for example, a hLF fragment lacking Gl ⁇ Arg " ).
- the hLF polypeptide fragments just described are chosen so that cysteine residue 10 (see SEQ ID NO: l ) is retained.
- the polypeptide includes a cysteine at approximately the same location in the sequence.
- Such a cysteine can be used to dimerize one fragment with another polypeptide having a cysteine residue, such as another hLF fragment, for example.
- dimerization can increase the activity of the polypeptide.
- the minimum length of the polypeptides may include more than 6 contiguous amino acids from the N-terminus of hLF.
- some polypeptides of the invention may include 7, 8, 9, 10. 1 1 or 12 contiguous amino acids from hLF, for example.
- the polypeptides may also include fewer than 27, 26 or 25 contiguous amino acids of hLF.
- the polypeptides of the invention may include less than 24. 22, 20, 18, 16. 14 or 12 contiguous amino acids from hLF, or any number of amino acids therebetween.
- the polypeptide includes no more than 19 contiguous amino acids from hLF.
- the polypeptide includes no more than 1 1 contiguous amino acids from hLF.
- polypeptides of the invention include hLF( 1 -1 1 ), hLF(2- 1 1 ) and hLF(3- 1 1 ).
- the polypeptides of the invention are relatively short, the polypeptides can be readily synthesized using known methods.
- the polypeptides can be synthesized by the well-known Merrifield solid-phase synthesis method in which amino acids are sequentially added to a growing chain. See Merrifield (1963), J. Am. Chem. Soc. 85:2149-2156; and Atherton et al., "Solid Phase Peptide Synthesis," IRL Press. London, (1989). Automatic peptide synthesizers are commercially available from numerous suppliers, such as Applied Biosystems, Foster City, California. Additional synthetic approaches for preparing the polypeptides of the invention are described in Example I below.
- Certain polypeptides of the invention can also be prepared via a reduction and proteolysis method.
- This approach begins with pepsin digestion of hLF according to known methods (see. e.g.. Bellamy, W. et al.. Biochim. Biophys. Ada. 1 121 : 130-136 (1992); and Tomita, M. et al.. J. Dairy Sci. 74:4137-4132 (1991)).
- the digested products are subsequently reduced and alkylated using standard reagents (e.g.
- polypeptides of the invention can be prepared using well- known recombinant techniques in which a nucleotide sequence encoding the polypeptide of interest is expressed in cultured cells such as described in Ausubel et al, Current Protocols in Molecular Biology, Greene Publishing and Wiley-Interscience, New York (1987) and in Sambrook et al., Molecular Cloning - A Laboratory Manual. 2nd ed., vols. 1 -3, Cold Spring Harbor Laboratory . Cold Spring Harbor, New York, 1989, both of which are incorporated herein by reference in their entirety. Also see, Kunkel, 1985, Proc. Nail. Acad. Sci.
- nucleic acids encoding the desired polypeptides are used in expression vectors.
- expression vector generally refers to nucleotide sequences that are capable of affecting expression of a gene in hosts compatible with such sequences. These expression vectors typically include at least suitable promoter sequences and optionally, transcription termination signals. Additional factors necessary or helpful in effecting expression may also be used as described herein.
- DNA encoding the polypeptides of the present invention will typically be incorporated into DNA constructs capable of introduction into and expression in an in vitro cell culture.
- DNA constructs will be suitable for replication in a prokaryotic host, such as bacteria, e.g. , E. coli, or may be introduced into a cultured mammalian, plant, insect, yeast, fungi or other eukaryotic cell lines.
- DNA constructs prepared for introduction into a particular host will typically include a replication system recognized by the host, the intended DNA segment encoding the desired polypeptide, and transcriptional and translational initiation and termination regulatory sequences operably linked to the polypeptide encoding segment.
- a DNA segment is
- a promoter or enhancer is operably linked to a coding sequence if it stimulates the transcription of the sequence.
- DNA for a signal sequence is operably linked to DNA encoding a polypeptide if it is expressed as a preprotein that participates in the secretion of the polypeptide.
- DNA sequences that are operably linked are contiguous, and, in the case of a signal sequence, both contiguous and in reading phase.
- enhancers need not be contiguous with the coding sequences whose transcription they control. Linking is accomplished by ligation at convenient restriction sites or at adapters or linkers inserted in lieu thereof.
- an appropriate promoter sequence generally depends upon the host cell selected for the expression of the DNA segment.
- suitable promoter sequences include prokaryotic, and eukaryotic promoters well-known in the art. See, e.g. , Sambrook et al., Molecular Cloning: A Laboratory Manual (2d ed.), vols. 1 -3, Cold Spring Harbor Laboratory (1989).
- the transcriptional regulatory sequences will typically include a heterologous enhancer or promoter which is recognized by the host.
- the selection of an appropriate promoter will depend upon the host, but promoters such as the trp, lac and phage promoters, tRNA promoters and glycolytic enzyme promoters are known and available. See Sambrook et al., supra.
- expression vectors which include the replication system and transcriptional and translational regulatory sequences together with the insertion site for the polypeptide encoding segment may be employed. Examples of workable combinations of cell lines and expression vectors are described in Sambrook et al., supra, and in Metzger et al. Nature 334:31-36 (1988).
- suitable expression vectors may be expressed in, e.g., insect cells, e.g., Sf9 cells, mammalian cells, e.g., CHO cells and bacterial cells, e.g., E. coli.
- the polypeptides of the invention are produced by expression in transgenic animals (i.e., non-human animals containing an exogenous DNA sequence in the genome of germ-line and somatic cells introduced by way of human intervention) such as bovines, goats, rabbits, sheep, pigs or mice.
- transgenic animals i.e., non-human animals containing an exogenous DNA sequence in the genome of germ-line and somatic cells introduced by way of human intervention
- transgenic animals i.e., non-human animals containing an exogenous DNA sequence in the genome of germ-line and somatic cells introduced by way of human intervention
- transgenic animals i.e., non-human animals containing an exogenous DNA sequence in the genome of germ-line and somatic cells introduced by way of human intervention
- Methods for production of recombinant polypeptides by transgenic nonhuman species are known in the art and are described, for example, in U.S. Patent Nos. 5,304,489; 5,633,076; and 5,565,362 which
- transgenic animals are the isolation of the polypeptides of interest in large amounts, especially by economical purification methods.
- transgenic bovine species containing a transgene encoding a human lactoferrin polypeptide targeted for expression in mammary secreting cells is described in WO 91/08216, incorporated herein by reference in its entirety.
- lactoferrin variants are produced in transgenic bovines the human protein typically is separated from the bovine milk proteins (e.g., whey proteins, caseins, bovine lactoferrin, IgA, albumin, lysozyme, ⁇ - lactoglobulin) before use (e.g., administration to patients).
- bovine milk proteins e.g., whey proteins, caseins, bovine lactoferrin, IgA, albumin, lysozyme, ⁇ - lactoglobulin
- use may be made of whole or partially purified bovine milk containing the desired polypeptide.
- Another method for preparing the polypeptides of the invention is to employ an in vitro transcription translation system.
- DNA encoding a polypeptide of the invention is cloned into an expression vector as described supra.
- the expression vector is then transcribed and translated in vitro.
- the translation product can be used directly or first purified.
- Polypeptides resulting from in vitro translation typically do not contain the post- translation modifications present on polypeptides synthesized in vivo.
- Methods for synthesis of polypeptides by in vitro translation are described by, for example, Berger & Kimmel, Methods in Enzymology, Volume 152, Guide to Molecular Cloning Techniques, Academic Press, Inc., San Diego, CA, 1987 (incorporated herein by reference in its entirety).
- the polypeptides and pharmaceutical compositions of the invention exhibit a number of biological activities that provide benefit in therapeutic or prophylactic applications.
- the compositions are useful in treating various microbial infections such as bacterial infections.
- the polypeptides and pharmaceutical compositions may also have various other beneficial activities. These include anti-inflammatory, anti-viral and anti-infective activities, as well a pro- and anti-coagulant effects, modulation of complement activation, inhibition of lipoylpolysaccharide-(LPS) mediated activation of neutrophils, and growth promotion of intestinal epithelial cells.
- Other properties and biological activities of lactoferrin are described in Nuijens et al., 1996, J. Mammary Gland Biology and Neoplasia 1 :3, 283-293, which is incorporated herein by reference in its entirety.
- Therapeutic indications for the pharmaceutical compositions described herein include use in therapy or prophylaxis of infection, including local infection, large scale
- compositions can also be used to prepare or treat organ transplant recipients or other immunosuppressed individuals (e.g.. AIDS patients) against the effects of infections.
- compositions are effective in treating a variety of microbial infections, such as various viral, bacterial and fungal infections.
- the compositions are effective in treating Gram-negative and Gram-positive bacteria.
- pathogenic bacteria causing infections treatable by methods of the invention include: Listeria, Escherichia, chlamydia, rickettsial bacteria, mycobacteria, staphylococci, treptocci, pneumonococci, meningococci and conococci, Klebsiella, proteus, serratia, pseudomonas, Legionel/a, diphtheria, salmonella, bacilli, cholera, tetanus, botulism, anthrax, plague, leptospirosis, and Lymes disease bacteria.
- pathogenic viruses causing infections treatable by methods of the invention include: hepatitis (A, B. or C), herpes virus (e.g., VZV, HSV-I, HAV-6, HSV-II, and CMV, Epstein Barr virus), adenovirus, influenza virus, flaviviruses. echovirus, rhinovirus, coxsackie virus, coronavirus.
- herpes virus e.g., VZV, HSV-I, HAV-6, HSV-II, and CMV, Epstein Barr virus
- adenovirus e.g., influenza virus, flaviviruses. echovirus, rhinovirus, coxsackie virus, coronavirus.
- RSV respiratory syncytial virus
- mumps virus mumps virus
- rotavirus measles virus
- rubella virus parvovirus
- vaccinia virus HTLV virus
- dengue virus papillomavirus
- molluscum virus poliovirus
- rabies virus JC virus
- arboviral encephalitis virus and human immunodeficiency virus (HIV virus; e.g., type I and II).
- pathogenic fungi causing infections treatable by methods of the invention include: Candida (e.g., albicans, krusei, glabrata, tropicalis), Cryptococcus neoformans, Aspergillus (e.g., fumigatus, niger), Genus Mucorales (Mucor, Absidia, Rhizopus), Sporothrix schenkii, Blastomyces dermatitidis, Paracoccidioides brasiliensis, Coccidioides immitis and Histoplasma capsulatum.
- Candida e.g., albicans, krusei, glabrata, tropicalis
- Cryptococcus neoformans e.g., Aspergillus (e.g., fumigatus, niger), Genus Mucorales (Mucor, Absidia, Rhizopus), Sporothrix schenkii, Blastomyces dermatitidis, Paracoccidioides brasiliensis,
- pathogenic parasites causing infections treatable by methods of the invention include: Entamoeba histolytica, Balantidium coli, Naegleria, Fowler i, Acanthamoeba sp., Giardia lambia, Cryptosporidium sp., Pneumocystis carinii,
- Plasmodium vivax Babesia microti, Trypanosoma brucei, Trypanosoma cruzi, Leishmania donovani, Toxoplasma gondii and Plasmodium falcipar is.
- the efficacy of the polypeptides and pharmaceutical compositions of the invention may be further improved by combining the use of the compositions of the invention with compositions for treating microbial infections known in the art per se.
- the polypeptides and compositions of the invention may be combined with e.g. penicillins such as amplicillin. cephalosporins, erythromycin, kanamycin, gentamicin. vancomycin or tetracyclines. for treating bacterial infections.
- penicillins such as amplicillin. cephalosporins, erythromycin, kanamycin, gentamicin. vancomycin or tetracyclines.
- antiviral nucleoside analogs such as aclvcovir. ganciclovir, zidovudine (AZT) or didanosine.
- the pol) peptides and compositions of the invention may be combined with amphotericin B, nystatin, miconazole, fluconazole, clotrimazole, terbinafine, naftifine or butenafine.
- the composition includes a polypeptide and a pharmaceutically acceptable excipient, wherein the polypeptide has a defined length, consists of one or more arginine residues at the N-terminus and has antimicrobial activity (e.g.. is effective in killing viruses or bacteria).
- the polypeptide includes a contiguous segment of up to 27 amino acids and the N-terminal amino acids of the polypeptide consists of the residues XRR (where X is any amino acid and R is arginine).
- the invention also includes compositions in which the polypeptide is no longer than 26 amino acids and the N-terminal amino acids are RR.
- the polypeptide is shorter, such as 5, 10, 15, 20 or 25 amino acids long, or any length therebetween.
- the polypeptide is even smaller in other compositions.
- the polypeptide may simply consist of the N-terminal XRR or RR residues.
- the polypeptides used in the pharmaceutical compositions can also include any of the polypeptides described above.
- the remaining contiguous amino acid sequence of the polypeptide sequence can vary so long as the polypeptide has antimicrobial activity.
- the remaining contiguous sequence can consist of a contiguous amino acid sequence from hLF, especially the sequence beginning after Arg 3 .
- the polypeptide can be hLF(l -l l ), hLF(2-l l ) or hLF(3-l l ).
- the amino acid sequence may include sequences wherein a small number (e.g.. one, tw or three) amino acids are inserted or removed from the hLF sequence.
- the polypeptide includes contiguous sequences from hLF, wherein one or more of the amino acids has been chemically modified.
- compositions may also include, depending on the formulation desired, pharmaceutically-acceptable.
- non-toxic carriers of diluents which are defined as vehicles commonly used to formulate pharmaceutical compositions for animal or human administration.
- the diluent is selected so as not to affect the biological activity of the combination. Examples of such diluents are distilled water, buffered water, physiological saline, PBS, Ringer's solution, dextrose solution, and Hank's solution.
- the pharmaceutical composition or formulation may also include other carriers, adjuvants, or non-toxic, nontherapeutic, nonimmunogenic stabilizers, excipients and the like.
- compositions may also include additional substances to approximate physiological conditions, such as pH adjusting and buffering agents, toxicity adjusting agents, wetting agents, detergents and the like. Because of the ability of hLF to bind iron, in some instances it may be beneficial to include iron in the pharmaceutical composition.
- the composition may also include any of a variety of stabilizing agents, such as an antioxidant for example.
- the polypeptides may be complexed with various well-known compounds that enhance the in vivo stability of the polypeptide, or otherwise enhance its pharmacological properties (e.g., increase the half-life of the polypeptide, reduce its toxicity, enhance solubility or uptake). Examples of such modifications or complexing agents include the production of sulfate, gluconate, citrate, phosphate and the like.
- the polypeptides of the composition may also be complexed with molecules that enhance their in vivo attributes. A list of such molecules, provided by way of example and not limitation, includes carbohydrates, polyamines, amino acids, other peptides, ions (e.g., sodium, potassium, calcium, magnesium, manganese), and lipids.
- compositions containing the polypeptides can be administered for prophylactic and/or therapeutic treatments.
- the polypeptide in the pharmaceutical composition typically is present in a therapeutic amount, which is an amount sufficient to remedy a disease state or symptoms, particularly symptoms associated with a microbial infection, or otherw ise prevent, hinder, retard, or reverse the progression of disease or infection or any other undesirable symptoms in any way whatsoever.
- the concentration of the polypeptide in the pharmaceutical composition can vary widely, i.e., from less than about 0.1% by weight, usually being at least about 1 % by weight, to as much as 20% by weight or more.
- compositions are administered to a patient already suffering from a disease, as just described, in an amount sufficient to cure or at least partially arrest the symptoms of the disease or infection and its complications.
- An appropriate dosage of the pharmaceutical composition or polypeptide of the invention is readily determined according to any one of several well-established protocols. For example, animal studies (e.g., mice, rats) are commonly used to determine the maximal tolerable dose of the bioactive agent per kilogram of weight. In general, at least one of the animal species tested is mammalian. The results from the animal studies can be extrapolated to determine doses for use in other species, such as humans for example.
- compositions containing the compounds of the invention are administered to a patient susceptible to or otherwise at risk of a particular disease or infection. Such an amount is defined to be a "prophylactically effective" amount or dose. In this use, the precise amounts again depends on the patient's state of health and weight. Typically, the dose ranges from about 1 to 500 mg of purified protein per kilogram of body weight, with dosages of from about 5 to 100 mg per kilogram being more commonly utilized.
- compositions described herein can be administered in a variety of different ways. Illustrative examples include administering a composition containing a pharmaceutically acceptable carrier via oral, intranasal, rectal, topical, intraperitoneal, intravenous, intramuscular, subcutaneous, subdermal, transdermal, intrathecal, and intracranial methods.
- the preferred formulation and delivery option typically varies depending upon the location and size of the area requiring treatment. For example, for localized infections, the formulation may be designed for topical application or localized injection, for example. Systemic reactions, in contrast, may be treated or prevented by administration of compositions formulated for parenteral administration.
- the active ingredient can be administered in solid dosage forms, such as capsules, tablets, and powders, or in liquid dosage forms, such as elixirs, syrups, and suspensions.
- the active component(s) can be encapsulated in gelatin capsules together with inactive ingredients and powdered carriers, such as glucose, lactose, sucrose, mannitol. starch, cellulose or cellulose derivatives, magnesium stearate. stearic acid, sodium saccharin, talcum, magnesium carbonate and the like.
- inactive ingredients examples include red iron oxide, silica gel, sodium lauryl sulfate, titanium dioxide, and edible white ink.
- Similar diluents can be used to make compressed tablets. Both tablets and capsules can be manufactured as sustained release products to provide for continuous release of medication over a period of hours. Compressed tablets can be sugar coated or film coated to mask any unpleasant taste and protect the tablet from the atmosphere, or enteric-coated for selective disintegration in the gastrointestinal tract.
- Liquid dosage forms for oral administration can contain coloring and flavoring to increase patient acceptance.
- solid or liquid formulations in an enteric-coated or otherwise protected form.
- the formulation can be mixed or simply coadministered with a protectant, such as a liquid mixture of medium chain triglycerides, or the formulation can be filled into enteric capsules (e.g., of soft or hard gelatin, which are themselves optionally additionally enteric coated).
- enteric capsules e.g., of soft or hard gelatin, which are themselves optionally additionally enteric coated.
- solid formulations comprising the polypeptide can be coated with enteric materials to form tablets.
- the thickness of enteric coating on tablets or capsules can be, for example, from 0.5 to 4 microns in thickness.
- the enteric coating may comprise any of the enteric materials conventionally utilized in orally administrable pharmaceutical formulations. Suitable enteric coating materials are known, for example, from Remington 's Pharmaceutical Sciences, Mace Publishing Company, Philadelphia, 17th ed. (1985); and Hagars Handbuch der Pharmazeutica Praxie, Springer Verlag, 4 ed., Vol. 7a (1971), both of which are incorporated herein by reference in their entirety.
- compositions e.g., liposomes. or other lipidic complexes
- lipid-associated structures e.g., liposomes. or other lipidic complexes
- the complex containing the composition may subsequently be targeted to specific target cells by the incorporation of appropriate targeting molecules (e.g., specific antibodies or receptors). It is also possible to directly complex the polypeptide with a targeting agent.
- compositions prepared for intravenous administration typically contain 100 to 500 ml of sterile 0.9% NaCl or 5% glucose optionally supplemented with a 20% albumin solution and 100 to 500 mg of a polypeptide of the invention.
- a typical pharmaceutical composition for intramuscular injection would be made up to contain, for example, 1 ml of sterile buffered water and 1 to 10 mg of the purified polypeptide of the invention.
- Methods for preparing parenterally administrable compositions are well-known in the art and described in more detail in various sources, including, for example, Remington's
- compositions are to be used in vivo
- the components used to formulate the pharmaceutical compositions of the present invention are preferably of high purity and are substantially free of potentially harmful contaminants (e.g., at least National Food (NF) grade, generally at least analytical grade, and more typically at least pharmaceutical grade).
- compositions intended for in vivo use are usually sterile.
- the resulting product is typically substantially free of any potentially toxic agents, particularly any endotoxins, which may be present during the synthesis or purification process.
- Compositions for parental administration are also sterile, substantially isotonic and made under GMP conditions.
- any of the above pharmaceutical compositions may in addition to the polypeptides of the invention comprise one or more further antimicrobial agents known in the art per se.
- the pharmaceutical compositions may, in addition to the polypeptides of the invention, comprise antibiotics such as amplicillin, cephalosporins, erythromycin, kanamycin, gentamicin. vancomycin and/or tetracyclines.
- antibiotics such as amplicillin, cephalosporins, erythromycin, kanamycin, gentamicin. vancomycin and/or tetracyclines.
- antiviral nucleoside analogs such as aclycovir, ganciclovir, zidovudine (AZT) and/or didanosine.
- compositions may, in addition to the polypeptides of the invention, comprise amphotericin B, nystatin, miconazole, fluconazole, clotrimazole, terbinafine, naftifine and/or butenafine.
- Methods for detecting microbial infections such as bacterial infections are also provided by the invention.
- the method typically involves administering a labeled polypeptide to a patient infected with, or suspected of being infected with, some type of microbial organism.
- the polypeptide is hLF or a fragment thereof which is capable of interacting with the organism. Because the labeled polypeptide is capable of interacting with the infectious organism, it accumulates at the site of infection. It is possible to detect the accumulation of the polypeptide at a site of infection using various detectors which are sensitive to the label that is attached to the polypeptide.
- the polypeptide used can vary, but includes the polypeptides of the invention described herein.
- hLF as well as other fragments (i.e., fragments other than the N-tcrminal fragments described above) which are capable of binding to microbial organisms such as bacteria can also be utilized.
- hLF( l -l 1 ), hLF(2-l 1 ) and hLF(3-l 1 ) rapidly accumulate at the site of infection.
- the label utilized to label the polypeptide can vary widely; the label simply needs to be a molecule or macromolecule that is capable of generating a detectable signal and that can be attached to the polypeptide.
- Illustrative examples of such molecules include radioactive isotopes, fluorophors, chromophores, electron dense reagents, magnetic particles, enzymes, and ligands having a specific binding partner (e.g., biotin).
- the detector used to detect the label can be any device which is capable of detecting the signal generated by the label.
- the detector can include a gamma camera. Using such a camera it is possible to obtain images of the site of infection which can be utilized in various research, diagnostic and therapeutic applications.
- the invention further provides methods for screening polypeptides to determine which of the polypeptides containing one or more amino acids from the first cationic domain of hLF but lacking residues from the second cationic domain have antimicrobial activity. Screening can be performed using in vitro or in vivo methods. In vitro methods generally involve mixing a suspension containing the bacteria of interest with a solution containing the test polypeptide. Typically, several different concentrations of the test polypeptide are tested. At various different time intervals, the number of viable bacteria are determined using standard microbiological techniques. Results are compared with control solutions which contain no polypeptide at all or contain a polypeptide known not to have antibacterial activity. Additional details regarding this approach are set forth in Example II below.
- the in vivo methods generally involve injecting a test animal (e.g., a mouse or rat) with a known amount of bacteria suspended in a test solution. Control animals are injected with a solution containing no polypeptide or a solution containing a polypeptide which is known to have no antimicrobial activity. The bacteria arc allowed to grow for a set period of time and then the test animals are sacrificed. Infected tissue is removed and the number of bacteria present in the infected tissue is determined using standard microbiological techniques. Example II below provides more specifics regarding one in vivo method.
- a test animal e.g., a mouse or rat
- Control animals are injected with a solution containing no polypeptide or a solution containing a polypeptide which is known to have no antimicrobial activity.
- the bacteria arc allowed to grow for a set period of time and then the test animals are sacrificed. Infected tissue is removed and the number of bacteria present in the infected tissue is determined using standard microbiological techniques. Example II
- polypeptides of the present invention have various activities associated with the first cationic cluster of hLF.
- the polypeptides of the invention are especially useful in selectively triggering responses involving the first cationic domain while avoiding the activation of responses associated with the binding of the second cationic domain and/or activities related to the iron binding activities of hLF.
- Some polypeptides of the invention can bind and neutralize heparin and lipopolysaccharide (LPS), lipid A, and DNA and human lysozyme. Certain polypeptides provided herein can also bind to various target cells. Lactoferrin binds to cell surfaces through two classes of LF binding sites: relatively low affinity sites which are cell surface sulfated molecules (e.g., cell surface proteoglycans or glycosaminoglycans) and high affinity- receptors. Since binding of LF to the low affinity sites involves the first cationic domain, some polypeptides of the present invention can selectively bind to the low affinity sites without activating the LF high affinity receptors.
- LPS heparin and lipopolysaccharide
- lipid A lipid A
- DNA and human lysozyme DNA and human lysozyme.
- Certain polypeptides provided herein can also bind to various target cells. Lactoferrin binds to cell surfaces through
- polypeptides of the present invention are useful in neutralizing heparin or lipopolysaccharide (LPS) without activating the lactoferrin high affinity receptor.
- LPS lipopolysaccharide
- certain polypeptides can neutralize the anticoagulant activity of heparin (including low molecular weight heparin).
- By neutralizing bacterial lipopolysaccharides certain polypeptides of the invention can reduce the inflammatory response associated with these compounds.
- Cell binding assays are well-known and are described in, e.g., Mazurier, 1989, Eur. J. Biochem. 179:481-87.
- Cells which the polypeptides can interact with include, intestinal cells (Hu et al., 1990, Biochemistry 29:535-541 ; Kawakam et al, 1991 , Am. J. Physiol. 26LG841-G846; Mikogami et al, 1994, Am. J. Physiol. 267:G308-G31), mammary gland epithelial cells (Rochard et al, 1992. Anticancer Res. 1 :2047-2052), hepatocytes (Regoeczi et al, 1985. Am. J. Physiol. 248:G8-G14; MacAbee et al, 1991 , J Biol. Chem.
- po peptides and compositions of the invention can also be used to inhibit entry into a cell of viruses, for example, CMV (cytomegalovirus), HIV (human immunodeficiency viruses) or HSV1 (herpes simplex virus 1) viruses.
- methods of the invention include administering a polypeptide of the invention to a patient to protect against infection by viruses. While not intending to be limited to this particular explanation, the antiviral action is thought to be mediated by interaction of hLF with cell surface proteoglycans (e.g.. heparin) employed by viral particles for cell entry, and/or by the stimulation of natural killer cells.
- polypeptides are also useful in reducing inflammation. This can occur, as noted above, through neutralization of bacterial lipopolysaccharides, as well as through a reduction in cytokine production and neutrophil degranulation.
- the invention provides methods in which some polypeptides and pharmaceutical compositions of the invention are administered to a patient to inhibit myelopoieses and reduce production of GM-CSF.
- the invention further provides antibodies which specifically bind to the polypeptides of the invention.
- Monoclonal antibodies are made from the polypeptides of the invention or from antigen-containing fragments thereof by methods that are well-known in the art (see, e.g., Kohler, et al. Nature, 256:495, (1975); and Hariow & Lane, Antibodies, A
- the antibodies of the invention are useful in purifying polypeptides of the invention, in screening cDNA expression libraries, and for identifying clones containing cDNA inserts which encode structurally-related, immunocrossreactive polypeptides.
- the following examples are presented to further illustrate certain aspects of the invention and are not to be construed so as to limit the scope of the invention.
- Natural hLF ( Mr 77,000) was purified from the fresh milk of a single donor by cationic exchange chromatography using S-Sepharose (14). The N-terminus of the protein was intact, as determined by N-terminal analytical Mono-S chromatography and N-terminal Edman degradation and sequencing. Human lactoferrin, free of contamination with endotoxin and human lysozyme (14) and pure by SDS PAGE, was dialyzed against saline and stored at -70 °C at concentrations of approximately 20 mg/ml.
- a hLF variant lacking the first three N-terminal residues (GRR), further referred to as hLF " was isolated from natural hLF treated with trypsin as described (18).
- Bovine serum albumin (BSA, Sigma Chem Co. St. Louis, MO) was applied instead of hLF or hLF ' N as a control.
- Synthetic peptides corresponding to residues 1 -1 1 of hLF (GRRRRSVQWCA (residues 1 -1 1 of SEQ ID NO: l -- SEQ ID NO: 2); Mr 1 ,374), further referred to as hLF(l -l 1 ), and fragments thereof, and a peptide corresponding to residues 21 -31 of hLF (FQ WQRNMRKVR (residues 21 -31 of SEQ ID NO:l -- SEQ ID NO: 4); Mr 1 ,548), referred to as hLF(21 -31), were prepared and purified as described (19).
- Antibiotic resistant Staphylococcus aureus strain 2141 a clinical isolate (Dept. of Infectious Diseases, LUMC), was resistant to a panel of antibiotics including methicillin and displayed only limited sensitivity to teicoplanin and rifampicin.
- Listeria monocytogenes strain EGD, Klebsiella pneumoniae (43816), and Escherichia coli 054 were purchased from the American Type Culture Collection (Rockville, MD). Antibiotic resistant S. aureus, K. pneumoniae and E.
- coli were cultured overnight in nutrient broth (Oxoid, Basingstoke, UK) at 37 °C, diluted in tryptase soya broth (TSB, Oxoid) and cultured for an additional 2 h in a shaking waterbath at 37 °C.
- TTB tryptase soya broth
- L. monocytogenes was cultured overnight in brain heart infusion broth (BHI, Oxoid ) at 37 °C. diluted and cultured for an additional 2 h in a shaking waterbath at 37 °C.
- A. Assay for Antibacterial Activity of Lactoferrins and Related Polypeptides An in vitro assay was used to assess the antibacterial activity of hLF and related peptides.
- bacteria were washed with phosphate-buffered saline (PBS) (pH 7.4) and diluted to a concentration of approximately 2x l 0 6 CFU/ml of phosphate-buffered saline (pH 6.0) supplemented with 0.1 % (v/v) Tween-20 (hereafter designated as PBS-Tw). Equal volumes of this bacterial suspension and PBS-Tw containing various concentrations of hLF or related synthetic peptides were mixed.
- PBS-Tw phosphate-buffered saline
- hLF "3N and related peptides e.g., hLF (1 - 1 1 ) and hLF(21 -31 ) in bacteria
- SFDA 5-sulfofluorescein diacetate
- FACS analysis 231
- Lactoferrins and related peptides were labeled with 99m Technetium ( m Tc) as described (22, 23). Briefly, 10 ⁇ l of a peptide solution (1 mg/ml of HAc) was added to 2 ⁇ l of an aseptic solution of 0.5 mg of stannous pyrophosphate/ml (Dept. of Clinical Pharmacy and Toxicology, Leiden University Medical Center (LUMC), Leiden. The Netherlands). Immediately thereafter, 4 ⁇ l of a solution of 10 mg of KBH (crystalline. Sigma) per ml of 0.1 M NaOH was added.
- KBH crystalline. Sigma
- mice Preceding scintigraphy, a subcutaneous injection of 0.1 ml, containing 0.2 mg diazepam (Valium, Hoffmann-La Roche, Mijdrecht, The Netherlands) was administered. Immediately thereafter, lxl 0 7 CFU of antibiotic resistant S aureus or K. pneumoniae in 0.1 ml of saline were injected into the right thigh muscle. Twenty-four hours thereafter, 0.2 ml of saline, containing various amounts of 99m Tc-hLF or radiolabeled peptides (0.3-30 ⁇ mol). were injected intravenously. As a control, mice were injected w ith 0.2 ml of saline containing BSA or saline without any peptide.
- mice were sacrificed by an intraperitoneal injection of 0.25 ml of saline containing 12 mg of sodium pentobarbital (Sanofi BV, Div. Algin, Maassluis, The Netherlands).
- the infected thigh muscles were removed and, after weighing, homogenized in 4 ml of PBS.
- Appropriate dilutions of the homogenates were applied onto diagnostic sensitivity test plates (Oxoid), and the number of colonies counted after an overnight culture at 37 °C. Results are expressed as the number of CFUs per gram of infected tissue. All negative cultures were assigned the values of 100 CFU/ml of homogenate, which is the lower limit of detection.
- anatomically adjusted regions of interest were drawn over the various organs and both thighs providing data about clearance of 99m Tc- peptides and accumulation at the site of infection.
- the clearance of 99m Tc-labeled hLF or related peptides from the circulation during the first hour after injection was assessed by determining the half-life (t ⁇ /2 ) of radioactivity in ROI drawn over the heart at various time intervals.
- the amount of radioactivity in the heart was corrected for decay and expressed as a percentage of injected dose (% ID).
- Accumulation of hLF and related peptides at the site of infection is expressed as the ratio of the counts in the infected (target) and the non-infected thigh muscle (non-target), further referred to as the T/NT ratio.
- hLF(21-31) which includes the second cationic domain of hLF
- hLF( 1-1 1) was then determined against L. monocytogenes, antibiotic resistant S. aureus and E. coli.
- hLF and hLF "3N al 4 °C had no affect on the number of viable bacteria nor on the non-specific esterase activity, whereas protegrin- 1 did.
- Protegrin-1 kills bacteria through another mechanism, i.e., membrane permeabilization.
- membrane permeabilization In contrast to natural hLF and hLF "3N , the fragments hLF(l -1 1 ), hLF(2-l 1), hLF(3-l 1) and hLF(4-l 1), but not hLF(5-l 1 ) and hLF(6-l 1), influenced the membrane permeability (FIGS. 3A-3C) in a dose- and time-dependent fashion.
- hLF antibacterial activity of hLF and various fragments thereof in treating infections initiated in mice was determined using the methods described in Example II.
- hLF "3N , hLF(l -l 1 ), hLF(4-l 1 ) or hLF(21 -31 ) the number of viable bacteria in mice with an infection with antibiotic resistant S. aureus was reduced in a dose-dependent fashion; maximum effects were seen with 1 nmole of hLF, 10 nmole of hLF "3N , 0.1 nmole of hLF(l -l 1 ) and 10 nmole of hLF(21 -31) (see FIG. 5).
- hLF and related peptides were compared with an established marker of infection/inflammation, (i.e., radiolabeled polyclonal human IgG (23)), for their usefulness in imaging a bacterial infection.
- an established marker of infection/inflammation i.e., radiolabeled polyclonal human IgG (23)
- the results clearly indicated that radiolabeled hLF and related peptides visualized the infection much faster than radiolabeled IgG.
- hLF(l-l 1) was also superior to the other peptides and hLF itself.
- hLF(l-l 1) was found to be much more effective than hLF(4-l 1) in reducing the number of viable bacteria in antibiotic-resistant S. aure us-infected mice.
- kidneys 1 1 ⁇ 1.7 14 ⁇ 0.7 14 ⁇ 0.4 18 ⁇ 9.9 bladder 40 ⁇ 0.3 29 ⁇ 3 4 22 ⁇ 5.6 38 ⁇ 2.3 liver 4 ⁇ 0.6 10 ⁇ 0.6 27 ⁇ 0.6 20 ⁇ 6.3
- mice infected with antibiotic resistant S aureus were injected with 1 nmole of y m technet ⁇ um-Iabelled hLF, hLF "1N , peptides hLF( 1 - 1 1 ), hLF(4- 1 1 ) or hLF(21 -31 ) and at various intervals thereafter the amount of radioactivity in the kidneys, bladder, liver and other organs was determined using a planar gamma camera equipped with a coll ⁇ nator using regions of interest drawn over the vai ious organs The amounts of radioactivit) are expressed as a percentage of the injected dose (% ID) Values are means and SD of three mice
- Lactoferrin-lipopolysaccharide interactions involvement of the 28-34 loop region of human lactoferrin in the high affinity binding of Escherichia coli 055B5 lipopolysaccharide. Biochem. J. 312:839-84.
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JP2001537352A JP2003521483A (en) | 1999-11-11 | 2000-11-10 | Antibacterial activity of the first cation cluster of human lactoferrin |
AT00981916T ATE313562T1 (en) | 1999-11-11 | 2000-11-10 | ANTIMICROBIAL EFFECT OF THE FIRST CATIONIC CLUSTER OF HUMAN LACTOFERRIN |
DE60025026T DE60025026T2 (en) | 1999-11-11 | 2000-11-10 | Antimicrobial action of the first cationic cluster of human lactoferrin |
CA002388910A CA2388910C (en) | 1999-11-11 | 2000-11-10 | Antimicrobial activity of the first cationic cluster of human lactoferrin |
AU19015/01A AU776044B2 (en) | 1999-11-11 | 2000-11-10 | Antimcrobial activity of the first cationic cluster of human lactoferrin |
US10/130,180 US7060677B1 (en) | 1999-11-11 | 2000-11-10 | Antimicrobial activity of the first cationic cluster of human lactoferrin |
EP00981916A EP1228097B1 (en) | 1999-11-11 | 2000-11-10 | Antimicrobial activity of the first cationic cluster of human lactoferrin |
CY20061100402T CY1105008T1 (en) | 1999-11-11 | 2006-03-21 | ANTIMICROBIAL ACTIVITY OF THE FIRST CATIONIC COMPLEX OF HUMAN LACTOFERRIN |
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Cited By (11)
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WO2001072322A2 (en) * | 2000-03-27 | 2001-10-04 | Pharming Intellectual Property B.V. | High dosage parenteral administration of lactoferrin |
WO2002042336A2 (en) * | 2000-11-21 | 2002-05-30 | The Texas A & M University System | Fgf-affinity chromatography |
EP1360961A1 (en) * | 2002-05-07 | 2003-11-12 | AM-Pharma B.V. | Use of antimicrobial peptides for potentiating the activity of antimicrobial agents |
WO2007055578A1 (en) * | 2005-11-11 | 2007-05-18 | Leids Universitair Medisch Centrum | Cyclic antimicrobial peptides derived from lactoferrin |
WO2008006125A1 (en) * | 2006-07-10 | 2008-01-17 | Österreichische Akademie der Wissenschaften | Antimicrobial peptides |
EP2030980A1 (en) * | 2007-08-28 | 2009-03-04 | AM-Pharma B.V. | Mutants of lactoferrin |
EP2060586A1 (en) * | 2007-11-14 | 2009-05-20 | PharmaSurgics in Sweden AB | New synthetic arginine substituted peptides and their use |
CN107304424A (en) * | 2016-05-23 | 2017-10-31 | 成都福际生物技术有限公司 | Transferrins DNA binding domain, recombinant DNA polymerase and the preparation method of transformation |
US11338020B2 (en) | 2018-01-09 | 2022-05-24 | Synthetic Biologics, Inc. | Alkaline phosphatase agents for treatment of neurodevelopmental disorders |
US11638699B2 (en) | 2018-03-20 | 2023-05-02 | Theriva Biologics, Inc. | Intestinal alkaline phosphatase formulations |
US11654184B2 (en) | 2018-03-20 | 2023-05-23 | Theriva Biologics, Inc. | Alkaline phosphatase agents for treatment of radiation disorders |
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AU2004273785A1 (en) * | 2003-07-16 | 2005-03-31 | The Research Foundation Of State University Of New York | Host defense factor X (HDFX) |
EP2050461A1 (en) | 2007-10-19 | 2009-04-22 | PharmaSurgics in Sweden AB | Peptides based on the sequence of human lactoferrin and their use |
EP2481751A1 (en) | 2011-01-26 | 2012-08-01 | PharmaSurgics in Sweden AB | Human lactoferrin derived peptides |
JP6099055B2 (en) * | 2014-05-09 | 2017-03-22 | アンスティチュ ナショナル ドゥ ラ サンテ エ ドゥ ラ ルシェルシュ メディカル | Fucoidan as a ligand for the diagnosis of degenerative lesions |
EP4137511A4 (en) * | 2020-04-16 | 2024-01-10 | Dermopartners, S.L. | New composition for use to treat and prevent infections by covid-19 and other coronaviruses |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0629347A1 (en) * | 1992-01-23 | 1994-12-21 | Morinaga Milk Industry Co., Ltd. | Antibacterial agent and treatment of article therewith |
US5861491A (en) * | 1994-02-16 | 1999-01-19 | Pharming B.V. | Isolation of lactoferrin from milk |
Family Cites Families (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP3506274B2 (en) * | 1994-09-01 | 2004-03-15 | 雪印乳業株式会社 | New peptides and immunostimulants |
JPH1192375A (en) * | 1997-09-25 | 1999-04-06 | Morinaga Milk Ind Co Ltd | Antibacterial agent capable of enhancing antibacterial activity of new quinolone-based antibacterial drug |
-
2000
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Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0629347A1 (en) * | 1992-01-23 | 1994-12-21 | Morinaga Milk Industry Co., Ltd. | Antibacterial agent and treatment of article therewith |
US5861491A (en) * | 1994-02-16 | 1999-01-19 | Pharming B.V. | Isolation of lactoferrin from milk |
Non-Patent Citations (5)
Title |
---|
DATABASE WPI Section Ch, Week 199621 Derwent Publications Ltd., London, GB; Class B04, AN 1996-205535 XP002166159 -& JP 08 073499 A (SNOW BRAND MILK PROD CO LTD), 19 March 1996 (1996-03-19) * |
DATABASE WPI Section Ch, Week 199924 Derwent Publications Ltd., London, GB; Class B02, AN 1999-283486 XP002166160 -& JP 11 092375 A (MORINAGA MILK IND CO LTD), 6 April 1999 (1999-04-06) * |
ELLISON R T III ET AL: "DAMAGE OF THE OUTER MEMBRANE OF ENTERIC GRAM-NEGATIVE BACTERIA BY LACTOFERRIN AND TRANSFERRIN" INFECTION AND IMMUNITY, vol. 56, no. 11, 1988, pages 2774-2781, XP000993092 ISSN: 0019-9567 * |
GROENINK JASPER ET AL: "Cationic amphipathic peptides, derived from bovine and human lactoferrins, with antimicrobial activity against oral pathogens." FEMS MICROBIOLOGY LETTERS, vol. 179, no. 2, 15 October 1999 (1999-10-15), pages 217-222, XP000997948 ISSN: 0378-1097 * |
NIBBERING,P.H ET AL.: "Human lactoferrin and peptides derived from its N-terminus are highly effective against infections with antibiotic-resistant bacteria" INFECTION AND IMMUNITY, vol. 69, no. 3, March 2001 (2001-03), pages 1469-1476, XP000993093 * |
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US11053479B2 (en) | 2016-05-23 | 2021-07-06 | Foregene Company Limited | Modified transferrin DNA binding domain, recombinant DNA polymerase and preparation method |
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