[go: up one dir, main page]

WO2008018084A2 - Peptides dérivés d'un inhibiteur de l'activateur du plasminogène pour prévenir un dommage neuronal - Google Patents

Peptides dérivés d'un inhibiteur de l'activateur du plasminogène pour prévenir un dommage neuronal Download PDF

Info

Publication number
WO2008018084A2
WO2008018084A2 PCT/IL2007/001007 IL2007001007W WO2008018084A2 WO 2008018084 A2 WO2008018084 A2 WO 2008018084A2 IL 2007001007 W IL2007001007 W IL 2007001007W WO 2008018084 A2 WO2008018084 A2 WO 2008018084A2
Authority
WO
WIPO (PCT)
Prior art keywords
amino acid
group
subject
eeiimd
peptide
Prior art date
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/IL2007/001007
Other languages
English (en)
Other versions
WO2008018084A3 (fr
Inventor
Abd Al-Roof Higazi
Douglas B. Cines
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Thrombotech Ltd
University of Pennsylvania Penn
Original Assignee
Thrombotech Ltd
University of Pennsylvania Penn
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 Thrombotech Ltd, University of Pennsylvania Penn filed Critical Thrombotech Ltd
Publication of WO2008018084A2 publication Critical patent/WO2008018084A2/fr
Anticipated expiration legal-status Critical
Publication of WO2008018084A3 publication Critical patent/WO2008018084A3/fr
Ceased legal-status Critical Current

Links

Classifications

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

Definitions

  • the present invention relates to therapeutic uses of peptides comprising six amino acid residues derived from amino acids 350-355 of human plasminogen activator inhibitor 1 (PAI-I) for preventing neuronal damage due particularly to hypoxic or thromboembolic stroke and brain injury.
  • PAI-I human plasminogen activator inhibitor 1
  • tPA tissue-type plasminogen activator
  • tPA may not be related exclusively to its proteolytic activity.
  • the inventor of the present application and co-workers have previously reported that exogenous tPA decreases cerebral vascular resistance in rats (Nassar et al., Blood 103: 897-902, 2004) and piglets (Armstead et al., J. Neurotrauma 21 : 1204-1211, 2004).
  • Studies have also demonstrated that the levels of tPA in the CNS are elevated after fluid percussion brain injury (FPI), thought to mimic concussive traumatic brain injury, and that administration of tPA at these concentrations induces cerebral vasodilation in naive animals (Armstead et al., Develop. Brain Res. 156: 139-146, 2005).
  • FPI fluid percussion brain injury
  • PAI plasminogen activator inhibitor
  • PAI-I plasminogen activator inhibitor
  • PAI-I derived peptide EEIIMD inhibit tPA-mediated signal transduction
  • Madison et al. Proc. Natl. Acad. Sci. U.S.A. 87: 3530-3533, 1990
  • the region comprises the sequence RMAPEEIIMDR.
  • U.S. Patent No. 6,750,201 to Cines et al. discloses compositions comprising peptides comprising at least six amino acids derived from PAI-I, particularly disclosed is a peptide of the sequence EEIIMD as set forth in SEQ ID NO:4, and methods of use thereof for promoting internalization and degradation of urokinase-type plasminogen activator.
  • WO 03/00642 discloses a six amino acid peptide having the amino acid sequence EEIIMD capable of reducing the undesirable side effects of fibrinolytic agents, for example, the risk of intracerebral hemorrhage in patients receiving tPA, uPA, tcuPA, streptokinase, rt-PA or alteplase, rt-PA derivatives or anisoylated streptokinase complex.
  • the peptide was introduced into the thrombolytic regimen in later stages to prevent the vasoactive or side effects of the primary thrombolytic agent.
  • WO 03/095476 to the inventor of the present invention teaches that administration of the peptide EEIIMD, or the peptide Ac-RMAPEEIIMDRPFLYVVR-Amide, anti-LRP antibodies or LRP antagonists, in combination with one or more fibrinolytic agents is useful for reducing the side effects due to vasoactivity caused by the fibrinolytic agent, and/or prolonging the half lives of the fibrinolytic agent.
  • WO 03/095476 further relates to combination compositions and/or therapy regimens comprising the polypeptide EEIIMD and/or the Ac- RMAPEEIIMDRPFL YVVR-amide peptide and one or more currently used plasminogen activators.
  • U.S. Patent Application Publication No. US2006/0069035 to the inventor of the present invention discloses that the EEIIMD peptide reduces the effective dosage of a thrombolytic agent required in the prevention or treatment of thromboembolic disorders.
  • the present invention provides uses of peptides consisting of six amino acid residues derived from amino acids 350-355 of human plasminogen activator inhibitor type 1 (PAI-I) for preventing neuronal damage in subjects suffering from or susceptible to neuronal damage.
  • PAI-I human plasminogen activator inhibitor type 1
  • the present invention now discloses for the first time that a 6-mer peptide of the amino acid sequence Ac-EEIIMD-amide as set forth in SEQ ID NO:1 corresponding to amino acids 350-355 of human PAI-I as set forth in SEQ ID NO:2 is capable of reducing brain edema and neurodegeneration after traumatic brain injury in pigs.
  • the neuroprotective effect of the peptide Ac-EEIIMD-amide was not associated with neutralizing the deleterious effects of exogenously administered tPA since the injured pigs were not treated with tPA.
  • the peptide Ac-EEIIMD-amide possesses unexpectedly neuroprotective activity irrespective of exogenous tPA and is therefore highly useful in the prevention of neuronal damage in subjects suffering from such damage while not being treated with tPA.
  • the present invention provides a method for preventing neuronal damage or the progression of neuronal damage in a subject suffering from or susceptible to such neuronal damage comprising administering to the subject a pharmaceutical composition comprising a therapeutically effective amount of an isolated peptide consisting of the amino acid sequence of general formula I: A-X 1 -X 2 -IIe-IIe-MCt-X 3 -B SEQ ID NO:3 wherein A is selected from the group consisting of hydrogen, acetyl, and an amino-terminal blocking group; X 1 is an amino acid selected from the group consisting of
  • X 2 is an amino acid selected from the group consisting of D and E
  • X 3 is an amino acid selected from the group consisting of D and E
  • B denotes a carboxylic acid, an amide, alcohol, ester, and a carboxyl-terminal blocking group, and pharmaceutically acceptable carrier, thereby preventing neuronal damage or the progression of neuronal damage in said subject.
  • the peptide consists of the amino acid sequence EEIIMD as set forth in SEQ ID NO:4. According to another embodiment, the peptide consists of the amino acid sequence Ac-EEIIMD-amide as set forth in SEQ ID NO: 1.
  • neuronal damage is due to an ischemic condition.
  • the ischemic condition is selected from the group consisting of ischemic or hypoxic stroke, hemorrhage, and traumatic brain injury.
  • neuronal damage is due to a non-ischemic condition.
  • the non-ischemic condition is selected from the group consisting of epilepsy, Alzheimer's disease, Huntington's disease, Down's syndrome, multiple sclerosis, and Parkinson's disease.
  • neuronal damage is due to a condition or disorder selected from the group consisting of spinal cord injury; perinatal hypoxia; ischemic condition subsequent to cardiac arrest; neurotrauma subsequent to cardiac bypass surgery or grafting; metabolically induced neurological damage; cerebral seizures; secondary neurodegenerative diseases (metabolic or toxic); memory disorders; and dementia including vascular dementia, multi-infarct dementia, Lewy body dementia, or neurogenerative dementia.
  • administering the pharmaceutical composition is performed by intravenous, subcutaneous, intramuscular, intraperitoneal, oral, topical, intradermal, intranasal, epidural, ophthalmic, or rectal route.
  • administering the pharmaceutical composition is performed by intravenous administration route.
  • the subject is a mammal.
  • the mammal is a human.
  • the pharmaceutical composition is formulated in a form selected from the group consisting of a pellet, a tablet, a capsule, a solution, a suspension, an emulsion, a gel, a cream, a suppository, a vaginal ring, and a depot.
  • the pharmaceutical composition is administered within twenty four hours after the subject experienced the neuronal damage due to an ischemic condition. According to a certain embodiment, the pharmaceutical composition is administered within six hours after the subject experienced the neuronal damage due to the ischemic condition.
  • the present invention provides a method for treating hypoxic or ischemic stroke in a subject comprising administering to the subject a pharmaceutical composition comprising a therapeutically effective amount of a peptide consisting of the amino acid sequence of general formula I according to the principles of the present invention and a pharmaceutically acceptable carrier, thereby treating the storke in said subject.
  • the peptide consists of the amino acid sequence EEIIMD as set forth in SEQ ID NO:4. According to another embodiment, the peptide consists of the amino acid sequence Ac-EEIIMD-amide as set forth in SEQ ID NO:1.
  • the pharmaceutical composition is administered within twenty four hours, alternatively within six hours after the subject experienced the stroke.
  • the present invention provides a method for treating brain injury in a subject comprising administering to the subject a pharmaceutical composition comprising a therapeutically effective amount of a peptide consisting of the amino acid sequence of general formula I according to the principles of the present invention and a pharmaceutically acceptable carrier, thereby treating the brain injury in said subject.
  • the peptide consists of the amino acid sequence EEIIMD as set forth in SEQ ID NO:4. According to another embodiment, the peptide consists of the amino acid sequence Ac-EEIIMD-amide as set forth in SEQ ID NO:1.
  • the present invention provides use of a peptide consisting of the amino acid sequence of general formula I according to the principles of the present invention for preventing neuronal damage or the progression of neuronal damage.
  • the present invention provides use of a peptide consisting of the amino acid sequence of general formula I according to the principles of the present invention for treating hypoxic or ischemic stroke.
  • the present invention provides use of a peptide consisting of the amino acid sequence of general formula I according to the principles of the present invention for treating brain injury.
  • FIG. 1 shows the neuroprotective effect of the PAI-I derived peptide Ac-EEIIMD- amide.
  • Mechanical occlusion of the MCA was performed in rats. Two hours after reperfusion was established, saline (Cont), tPA, tPA together with Ac-EEIIMD-amide, tPA together with Ac-EEIIMR-amide, or Ac-EEIIMD-amide alone was injected intravenously and the size of the infarct was measured twenty four hours later.
  • FIG. 2 shows brain sections of mice treated with saline (Control), saline containing tPA, or saline containing tPA and Ac-EEIIMD-amide two hours after mechanical occlusion of the MCA. Three anatomically parallel sequential brain sections are shown.
  • FIG. 3 shows that the PAI-I derived peptide does not accelerate tPA clearance.
  • 125 RPA was injected intravenously (i.v.) in rats alone or together with Ac-EEIIMD-amide. Blood samples were taken 1, 4, 8 and 20 min after injection, the plasma was separated and the radioactivity was measured.
  • FIG. 4 shows that Ac-EEIIMD-amide does not inhibit the fibrinolytic activity of tPA. Rats were given an intravenous injection of saline (Cont), tPA, tPA plus Ac- EEIIMD-amide, or Ac-EEIIMD-amide alone.
  • FIG. 5 shows that Ac-EEIIMD-amide reduces infarct size in animals treated with tPA after embolic stroke.
  • Microemboli were injected into the MCA. Two hours later, the animals were injected i.v. with saline (Cont.), tPA, tPA together with Ac-EEIIMD-amide, or Ac-EEIIMD-amide alone. Infarct size was measured twenty four hours later. Infarction induced by mechanical occlusion as described in FIG. 1 was determined in parallel and was designated 100%.
  • FIGs. 6A-B show that Ac-EEIIMD-amide decreases hippocampal and cortical histopathologic changes associated with fluid percussion brain injury (FPI).
  • FIG. 6A shows H & E stained brain sections from the CA3 hippocampus and parietal cortex of piglets 4 hrs after injury in sham, FPI and FPI + Ac-EEIIMD-amide.
  • FIG. 6B shows FluoroJade stained brain sections from the CA3 hippocampus and parietal cortex of sham, FPI + Ac-EEIIMD-amide, and FPI piglets. Scale bar is 50 um.
  • FIGs. 7A-B show that Ac-EEIIMD-amide decreases the number of degenerating neurons in the cortex.
  • FIG. 7A-B show that Ac-EEIIMD-amide decreases the number of degenerating neurons in the cortex.
  • FIG. 7A shows the number of degenerating neurons in the cortex of sham, FPI + Ac-EEIIMD-amide pretreated, and FPI animals.
  • FIG. 7B shows the number of degenerating neurons in the CA3 hippocampi of sham, FPI + Ac-EEIIMD-amide pretreated, and FPI animals.
  • FIGs. 8A-B show that Ac-EEIIMD-amide decreased the effect of FPI on swelling of cortex.
  • FIG. 8A shows photomicrographs of H & E stained cortex in sham animal (top) compared to FPI animals or FPI animals treated with Ac-EEIIMD-amide. The inset portion of the figure is 2OX magnification.
  • FIG. 8B shows the extent of tissue edema in sham, FPI + Ac-EEIIMD-amide pretreated, and FPI animals. L is left (ipsilateral, injured) side and R is right (contralateral) side.
  • FIG. 9. shows that Ac-EEIIMD-amide decreases post FPI brain water content. Brain water content was determined in sham animals (Control) or 4 hours after FPI. The animals were exposed to isoproterenol (Iso), fluid percussion brain injury (FPI), FPI + Ac- EEIIMD-amide, FPI + tP A, FPI + Iso, FPI + tPA + Ac-EEIIMD-amide, or FPI + Iso + Ac- EEIIMD-amide.
  • Iso isoproterenol
  • FPI fluid percussion brain injury
  • FPI + Ac- EEIIMD-amide FPI + tP A
  • FPI + Iso FPI + tPA + Ac-EEIIMD-amide
  • FPI + Iso + Ac- EEIIMD-amide FPI + Iso + Ac- EEIIMD-amide.
  • the present invention is based in part on the findings that a 6-mer peptide of the amino acid sequence acetyl (Ac)-EEIIMD-amide as set forth in SEQ ID NO:1 corresponding to amino acids 350-355 of human PAI-I as set forth in SEQ ID NO: 2 was capable of abolishing not only the tPA-induced increase in brain infarct size in mechanical models of stroke in rats or in rats in which stroke was induced by embolism and thereafter treated with tPA, but also was capable of reducing brain edema and neurodegeneration after traumatic brain injury in pigs that were not subjected to any treatment with exogenous tPA.
  • the present invention provides methods for preventing neuronal damage or the progression of neuronal damage in a patient suffering from or susceptible to such neuronal damage comprising administering a therapeutically effective amount of an isolated peptide comprising the amino acid sequence of general formula I: A-Xi-X 2 -Ile-Ile-Met-X 3 -B SEQ ID NO:3 wherein A is selected from the group consisting of hydrogen, acetyl, and an amino-terminal blocking group; X 1 is an amino acid selected from the group consisting of D, E, and R; X 2 is an amino acid selected from the group consisting of D and E; X 3 is an amino acid selected from the group consisting of D and E; and B denotes a carboxylic acid, an amide, alcohol, ester, and a carboxyl-terminal blocking group, and pharmaceutically acceptable carrier.
  • A is selected from the group consisting of hydrogen, acetyl, and an amino-terminal blocking group
  • X 1 is an amino acid selected from the group consist
  • the peptide comprises the amino acid sequence EEIIMD as set forth in SEQ ID NO:4 corresponding to amino acids 350-355 of mature human PAI-I as set forth in SEQ ID NO:2.
  • the peptide consists of the amino acid sequence Ac-EEIIMD-amide as set forth in SEQ ID NO:1. It is to be understood that the amino acid sequence EEIIMD corresponds to positions 373-378 of the amino acid sequence of human PAI-I which includes the signal peptide.
  • the present invention further provides methods for treating hypoxic or ischemic stroke in a subject and methods for treating brain injury in a subject, the methods comprise a step of administering a therapeutically effective amount of an isolated peptide comprising the amino acid sequence of general formula I: A-Xj-X 2 -Ile-Ile-Met-X 3 -B SEQ ID NO:3 wherein A is selected from the group consisting of hydrogen, acetyl, and an amino-terminal blocking group; Xi is an amino acid selected from the group consisting of
  • X 2 is an amino acid selected from the group consisting of D and E
  • X 3 is an amino acid selected from the group consisting of D and E
  • B denotes a carboxylic acid, an amide, alcohol, ester, and a carboxyl-terminal blocking group, and pharmaceutically acceptable carrier.
  • peptide as used throughout the specification and claims designates a linear series of amino acid residues connected one to the other by peptide bonds. The amino acid residues are represented throughout the specification and claims by one-letter or three-letter codes according to IUPAC conventions.
  • amino acid or “amino acid residue” is understood to include the 20 naturally occurring amino acids.
  • the peptides of the present invention comprise six amino acid residues.
  • the peptides of the present invention can be isolated by any protein purification method known in the art.
  • PAI-I can be subjected to one or more proteolytic enzymes to yield a mixture of peptides, which can further be purified by any protein purification method known in the art to obtain the isolated peptides.
  • PAI-I can be cleaved by chemical agents such as, for example, CNBr to yield a mixture of peptides that can be further purified to obtain isolated peptides.
  • the peptides of the present invention can also be prepared by methods well known in the art including chemical synthesis or recombinant DNA technology.
  • a preferred method of synthesizing the peptides of the present invention involves solid-phase peptide synthesis utilizing a solid support as described by Merrifield (see J. Am. Chem. Soc, 85:2149, 1964). Large-scale peptide synthesis is described, for example, by Andersson et al. (Biopolymers 55(3): 227-50, 2000). Examples of solid phase peptide synthesis methods include the BOC method, which utilizes tert-butyloxcarbonyl as the ⁇ - amino protecting group, and the FMOC method, which utilizes 9- fluorenylmethyloxcarbonyl to protect the ⁇ -amino of the amino acid residues, both methods are well-known by those of skill in the art. Alternatively, the peptides of the present invention can be synthesized by standard solution synthesis methods (see, for example, Bodanszky, M., Principles of Peptide Synthesis, Springer- Verlag, 1984).
  • the peptides according to the principles of the invention need not be identical to the amino acid sequence EEIIMD set forth in SEQ ID NO:4 so long as each peptide is capable of preventing or inhibiting neuronal damage.
  • the term “analog” includes any peptide comprising altered sequence by amino acid substitutions, additions, deletions, or chemical modifications of the peptides listed herein above and which displays neuroprotective activity.
  • amino acid substitutions it is meant that functionally equivalent amino acid residues are substituted for residues within the sequence resulting in a silent change.
  • one or more amino acid residues within the sequence can be substituted by another amino acid of a similar polarity, which acts as a functional equivalent, resulting in a silent alteration.
  • Substitutes for an amino acid within the sequence may be selected from other members of the class to which the amino acid belongs.
  • the non-polar (hydrophobic) amino acids include alanine, leucine, isoleucine, valine, proline, phenylalanine, tryptophan and methionine.
  • the polar neutral amino acids include glycine, serine, threonine, cysteine, tyrosine, asparagine, and glutamine.
  • the positively charged (basic) amino acids include arginine, lysine and histidine.
  • the negatively charged (acidic) amino acids include aspartic acid and glutamic acid. Such substitutions are known as conservative substitutions.
  • the present invention encompasses peptide analogs, wherein at least one amino acid is substituted by another amino acid to produce a peptide analog having increased stability or longer half-life as compared to the peptides listed herein above.
  • amino acid residues of the peptide sequences set forth herein above are all in the "L” isomeric form
  • residues in the "D” isomeric form can substitute any L-amino acid residue so long as the peptide analog retains neuroprotective activity.
  • Production of a retro-inverso D-amino acid peptide analog where the peptide is made with the same amino acids as disclosed, but at least one amino acid, and perhaps all amino acids are D-amino acids is well known in the art.
  • the result is an analog having the same structural groups being at the same positions as in the L-amino acid form of the peptide.
  • the peptide analog is more stable to proteolytic degradation and is therefore useful in many of the applications recited herein.
  • the present invention further encompasses peptide derivatives of the peptides listed herein above.
  • derivative refers to a peptide having an amino acid sequence that comprises the amino acid sequence of the peptide of the invention, in which one or more of the amino acid residues is subjected to chemical derivatizations by a reaction of side chains or functional groups, where such derivatizations do not destroy the neuroprotective activity of the peptide derivative.
  • Chemical derivatization of amino acid residues include, but are not limited to, acetylation, amidation, glycosylation, oxidation, reduction, myristylation, sulfation, acylation, ADP-ribosylation, cyclization, disulfide bond formation, hydroxylation, iodination, and methylation.
  • the present invention also encompasses those peptides in which free amino groups have been derivatized to form amine hydrochlorides, p-toluene sulfonyl groups, carbobenzoxy groups, t-butyloxycarbonyl groups, chloroacetyl groups or formyl groups.
  • Free carboxyl groups may be derivatized to form, for example, salts, methyl and ethyl esters or other types of esters, amides, alcohols, or hydrazides.
  • Free hydroxyl groups may be derivatized to form, for example, o-acyl or o-alkyl derivatives.
  • the imidazole nitrogen of histidine may be derivatized to form N-im-benzylhistidine.
  • peptides which contain one or more naturally occurring amino acid derivatives of the twenty standard amino acid residues.
  • 4-hydroxyproline may be substituted for proline
  • 5-hydroxylysine may be substituted for lysine
  • 3-methylhistidine may be substituted for histidine
  • homoserine may be substituted or serine
  • ornithine may be substituted for lysine.
  • the peptides may also contain non-natural amino acids.
  • non-natural amino acids are norleucine, ornithine, citrulline, diaminobutyric acid, homoserine, isopropyl Lys, 3-(2'- naphtyl)-Ala, nicotinyl Lys, amino isobutyric acid, and 3-(3'-pyridyl-Ala).
  • the peptides may also contain non-protein side chains.
  • the peptides of the present invention may also include one or more non-amino acid monomers (e.g., fatty acids, complex carbohydrates, and the like).
  • Peptides of the present invention also include any peptide having one or more additions of amino acid residues relative to the sequences of the peptides listed herein above, so long as the requisite neuroprotective activity is maintained.
  • the amino acid residues may be added at the amino terminus and/or carboxy terminus and/or along the peptide sequence.
  • linker by which the peptides of this invention can be conveniently bound to a carrier.
  • linkers are usually of at least one amino acid residue and can be of 40 or more residues, more often of 1 to 10 residues.
  • Typical amino acid residues used for linking are tyrosine, cysteine, lysine, glutamic and aspartic acid, or the like.
  • the present invention includes conjugates of the peptides of the invention.
  • conjugates are meant to define a peptide of the present invention coupled to or conjugated with another protein or polypeptide. Such conjugates may have advantages over the peptides themselves. Such conjugates can be made by protein synthesis, e. g., by use of a peptide synthesizer, or by ligating the appropriate nucleic acid sequences encoding the desired amino acid sequences to each other by methods known in the art, in the proper coding frame, and expressing the chimeric protein by methods commonly known in the art.
  • a peptide of the present invention may also be conjugated to itself or aggregated in such a way as to produce a large complex containing the peptide.
  • compositions and administration routes The present invention provides methods for preventing neuronal damage or the progression of neuronal damage in a subject suffering from or susceptible to such neuronal damage comprising administering to the subject a pharmaceutical composition comprising a therapeutically effective amount of a peptide of the invention and a pharmaceutically acceptable carrier.
  • pharmaceutical composition refers to a preparation of one or more of the peptides described herein with other chemical components such as pharmaceutically acceptable carriers and excipients.
  • the purpose of a pharmaceutical composition is to facilitate administration of an active ingredient to an organism.
  • pharmaceutically acceptable means approved by a regulatory agency of the Federal or a state government or listed in the U. S. Pharmacopeia or other generally recognized pharmacopeia for use in animals, and more particularly in humans.
  • compositions of the present invention can be formulated as pharmaceutically acceptable salts of the peptides of the present invention.
  • pharmaceutically acceptable salts refers to salts prepared from pharmaceutically acceptable non-toxic bases or acids including inorganic or organic bases and inorganic or organic acids. Salts derived from inorganic bases include aluminum, ammonium, calcium, copper, ferric, ferrous, lithium, magnesium, manganic salts, manganous, potassium, sodium, zinc, and the like. Particularly preferred are the ammonium, calcium, magnesium, potassium, and sodium salts.
  • Salts derived from pharmaceutically acceptable organic non- toxic bases include salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins, such as arginine, betaine, caffeine, choline, N,N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethyl-morpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine, methylglucamine, mo ⁇ holine, piperazine, piperidine, polyamine resins, procaine, purines, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, and the like.
  • basic ion exchange resins such as
  • salts may be prepared from pharmaceutically acceptable non-toxic acids, including inorganic and organic acids.
  • acids include acetic, benzenesulfonic, benzoic, camphorsulfonic, citric, ethanesulfonic, fumaric, gluconic, glutamic, hydrobromic, hydrochloric, isethionic, lactic, maleic, malic, mandelic, methanesulfonic, mucic, nitric, pamoic, pantothenic, phosphoric, succinic, sulfuric, tartaric, p-toluenesulfonic acid, and the like.
  • Particularly preferred are citric, hydrobromic, hydrochloric, maleic, phosphoric, sulfuric, and tartaric acids.
  • carrier refers to a diluent or vehicle that does not cause significant irritation to an organism and does not abrogate the biological activity and properties of the administered compound.
  • An adjuvant is included under these phrases.
  • Such pharmaceutical carriers can be sterile liquids, such as water and oils, including those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil and the like, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents. Water is a preferred carrier when the pharmaceutical composition is administered intravenously. Saline solutions and aqueous dextrose and glycerol solutions can also be employed as liquid carriers, particularly for injectable solutions.
  • compositions of the invention can further comprise an excipient.
  • excipient refers to an inert substance added to a pharmaceutical composition to further facilitate administration of an active ingredient.
  • suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene glycol, water, ethanol and the like.
  • the composition if desired, can also contain minor amounts of wetting or emulsifying agents, or pH buffering agents such as acetates, citrates or phosphates.
  • Antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; and agents for the adjustment of tonicity such as sodium chloride or dextrose are also envisioned.
  • compositions of the present invention can be manufactured by processes well known in the art, e.g., by means of conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping, or lyophilizing processes.
  • compositions which contain peptides as active ingredients are prepared as injectable, either as liquid solutions or suspensions, however, solid forms, which can be suspended or solubilized prior to injection, can also be prepared.
  • compositions can also take the form of emulsions, tablets, capsules, gels, syrups, slurries, powders, creams, depots, sustained-release formulations and the like.
  • Methods of introduction of a pharmaceutical composition comprising a peptide of the invention include, but are not limited to, intravenous, subcutaneous, intramuscular, intraperitoneal, oral, topical, intradermal, intranasal, epidural, ophthalmic, and rectal routes.
  • the pharmaceutical compositions can be administered by any convenient route, for example by infusion or bolus injection, by absorption through epithelial linings (e.g., oral mucosa, rectal and intestinal mucosa, etc.), and may be administered together with other therapeutically active agents.
  • the administration may be localized, or may be systemic.
  • compositions of the invention may be formulated in conventional manner using one or more physiologically acceptable carriers comprising excipients and auxiliaries, which facilitate processing of the active ingredients into preparations that can be used pharmaceutically. Proper formulation is dependent upon the route of administration chosen.
  • the active ingredients of the pharmaceutical composition may be formulated in aqueous solutions, preferably in physiologically compatible buffers such as Hank's solution, Ringer's solution, or physiological salt buffer.
  • physiologically compatible buffers such as Hank's solution, Ringer's solution, or physiological salt buffer.
  • penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art.
  • the pharmaceutical composition can be formulated readily by combining the active ingredients with pharmaceutically acceptable carriers well known in the art. Such carriers enable the pharmaceutical composition to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions, and the like, for oral ingestion by a patient.
  • Pharmacological preparations for oral use can be made using a solid excipient, optionally grinding the resulting mixture, and processing the mixture of granules, after adding suitable auxiliaries as desired, to obtain tablets or dragee cores.
  • Suitable excipients are, in particular, fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol; cellulose preparations such as, for example, maize starch, wheat starch, rice starch, potato starch, gelatin, gum tragacanth, methyl cellulose, hydroxypropylmethyl-cellulose, and sodium carbomethylcellulose; and/or physiologically acceptable polymers such as polyvinylpyrrolidone (PVP).
  • disintegrating agents such as cross-linked polyvinyl pyrrolidone, agar, or alginic acid or a salt thereof, such as sodium alginate, may be added.
  • Dragee cores are provided with suitable coatings.
  • suitable coatings For this purpose, concentrated sugar solutions may be used which may optionally contain gum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethylene glycol, titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures.
  • Dyestuffs or pigments may be added to the tablets or dragee coatings for identification or to characterize different combinations of active compound doses.
  • compositions that can be used orally include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol.
  • the push-fit capsules may contain the active ingredients in admixture with filler such as lactose, binders such as starches, lubricants such as talc or magnesium stearate, and, optionally, stabilizers.
  • the active ingredients may be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols.
  • stabilizers may be added. All formulations for oral administration should be in dosages suitable for the chosen route of administration.
  • the compositions may take the form of tablets or lozenges formulated in conventional manner.
  • the active ingredients for use according to the present invention are conveniently delivered in the form of an aerosol spray presentation from a pressurized pack or a nebulizer with the use of a suitable propellant, e.g., dichlorodifluoromethane, trichlorofluoromethane, dichloro-tetrafluoroethane, or carbon dioxide.
  • a suitable propellant e.g., dichlorodifluoromethane, trichlorofluoromethane, dichloro-tetrafluoroethane, or carbon dioxide.
  • the dosage may be determined by providing a valve to deliver a metered amount.
  • Capsules and cartridges of, for example, gelatin for use in a dispenser may be formulated containing a powder mix of the compound and a suitable powder base, such as lactose or starch.
  • compositions described herein may be formulated for parenteral administration, e.g., by bolus injection or continuous infusion.
  • Formulations for injection may be presented in unit dosage form, e.g., in ampoules or in multidose containers with, optionally, an added preservative.
  • the compositions may be suspensions, solutions, or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing, and/or dispersing agents.
  • compositions for parenteral administration include aqueous solutions of the active preparation in water-soluble form. Additionally, suspensions of the active ingredients may be prepared as appropriate oily or water-based injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, or synthetic fatty acid esters such as ethyl oleate, triglycerides, or liposomes. Aqueous injection suspensions may contain substances that increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol, or dextran. Optionally, the suspension may also contain suitable stabilizers or agents that increase the solubility of the active ingredients, to allow for the preparation of highly concentrated solutions.
  • the active ingredient may be in powder form for constitution with a suitable vehicle, e.g., a sterile, pyrogen-free, water-based solution, before use.
  • a suitable vehicle e.g., a sterile, pyrogen-free, water-based solution
  • compositions of the present invention may also be formulated in rectal compositions such as suppositories or retention enemas, using, for example, traditional binders and carriers such as triglycerides, microcrystalline cellulose, gum tragacanth or gelatin.
  • composition of the invention may be administered locally to the area in need of treatment; this may be achieved by, for example, and not by way of limitation, local infusion during surgery, topical application, e.g., in conjunction with a wound dressing after surgery, by injection, by means of a catheter, by means of a suppository, or by means of an implant, said implant being of a porous, non-porous, or gelatinous material.
  • Administration can also be by direct injection e.g., via a syringe, at the site of injury.
  • the pharmaceutical composition may be in the form of tablets or capsules, which can contain any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose; a disintegrating agent such as alginic acid, Primogel, or corn starch; a lubricant such as magnesium stearate or Sterotes; or a glidant such as colloidal silicon dioxide.
  • a binder such as microcrystalline cellulose, gum tragacanth or gelatin
  • an excipient such as starch or lactose
  • a disintegrating agent such as alginic acid, Primogel, or corn starch
  • a lubricant such as magnesium stearate or Sterotes
  • a glidant such as colloidal silicon dioxide.
  • dosage unit form can contain, in addition to materials of the above type, a liquid carrier such as a fatty oil.
  • dosage unit forms can contain other materials which modify
  • a peptide of the invention can be delivered in a controlled release system.
  • the peptide can be administered in combination with a biodegradable, biocompatible polymeric implant, which releases the peptide over a controlled period of time at a selected site.
  • a biodegradable, biocompatible polymeric implant which releases the peptide over a controlled period of time at a selected site.
  • preferred polymeric materials include polyanhydrides, polyorthoesters, polyglycolic acid, polylactic acid, polyethylene vinyl acetate, copolymers and blends thereof (See, Medical applications of controlled release, Langer and Wise (eds.), 1974, CRC Pres., Boca Raton, FIa.).
  • a controlled release system can be placed in proximity of the therapeutic target, thus requiring only a fraction of a systemic dose.
  • compositions suitable for use in the context of the present invention include compositions wherein the active ingredients are contained in an amount effective to achieve the intended purpose. More specifically, a "therapeutically effective amount” means an amount of a peptide effective to prevent, alleviate, or ameliorate symptoms of a condition or disease associated with neuronal damage in the subject being treated. Uses of the peptides
  • the present invention provides uses of the peptides of the invention for the treatment, prophylaxis and/or inhibition of neuronal damage in a subject in need thereof, for the treatment of hypoxic or ischemic stroke, and/or for the treatment of brain injury.
  • prophylactic use of the pharmaceutical compositions comprises administering to a subject in need thereof the pharmaceutical composition to prevent the onset of neurological damage; and to prevent the progression of neurological damage.
  • the peptides of the present invention have neuroprotective activity.
  • neuroprotective activity refers to prevention of onset of neuronal damage or arresting or inhibition of progression of neuronal damage in a subject.
  • neurode damage includes, but is not limited to, brain infarct, brain edema, neurodegeneration, and hemorrhage.
  • the treatment of the present invention can be applied to a variety of acute and chronic conditions.
  • the present invention can be used for the treatment of ischemic conditions, for example cerebral ischemia (thromboembolic or hypoxic or ischemic stroke, hemorrhage or brain injury as a result of trauma) which involve various forms of brain damage and may lead to acute or delayed damage to the brain neurons, and to neurodegeneration— for example after head trauma.
  • the present invention can be applicable to the treatment of relatively long-term neurodegeneration of non-ischemic origin (e.g., epilepsy, Alzheimer's disease, Huntington's disease, Downs syndrome, Multiple Sclerosis and Parkinson's disease) and neurological damage resulting from chronic infection, for example HIV producing the syndrome of AIDS.
  • Other conditions which can cause neural damage are well-known to an ordinarily skilled neurologist or similar physician and include: primary neurogenerative disease; spinal cord lesions; hypoxic processes such as perinatal hypoxia or ischemic processes such as subsequent to cardiac arrest; neurotrauma such as subsequent to cardiac bypass surgery or grafting; metabolically induced neurological damage; cerebral seizures; secondary neurodegenerative diseases (metabolic or toxic); memory disorders; vascular dementia, multi-infarct dementia, Lewy body dementia, or neurogenerative dementia.
  • the time of treatment is also significant and can be important. Administration can be before or after neuronal damage has occurred or is suspected. Administration before neuronal damage has occurred can be of value for prophylactic treatment, for example when the subject is considered to be at risk of an ischemic condition. Such conditions could be, for example in cardiac bypass surgery, in which a significant proportion of patients can suffer minor cerebral damage, or in childbirth, in which the fetus may be liable to problems in the fetal circulation potentially leading to anoxia and cerebral palsy and the like.
  • the subject is a mammal.
  • the mammal is a human.
  • a therapeutically effective amount is well within the capability of those skilled in the art, especially in light of the detailed disclosure provided herein.
  • the precise amount of the peptide administered to a particular subject, preferably a mammal, more preferably a human being, in the method of treatment of the present invention will depend on a number of factors, for example the specific peptide administered; its mode of administration and/or the use for which it is intended; the particular clinical condition being treated and/or its severity; and/or the age, body mass and/or past clinical history of the patient to be treated, and always lies within the sound discretion of the person administering and/or supervising the treatment, for example a medical practitioner such as nurse and/or physician.
  • a suitable daily dose of the peptide for administration to a mammal is generally from about 0.01 mg/day per kg of the mammal's body mass to about 80 mg/kg/day, more usually 0.2-40 mg/kg/day given in a single dose and/or in divided doses at one or more times during the day.
  • the pharmaceutical composition can contain from about 0.1% to about 99% by weight of the peptide and is generally prepared in unit dose form, a unit dose of a peptide generally being from about 0.1 mg to about 500 mg.
  • Dosage amount and administration intervals may be adjusted individually to provide sufficient plasma or local levels of the peptide to induce a neuroprotective effect.
  • dosing can be of a single or a plurality of administrations, with course of treatment lasting from several days to several weeks, or until cure is effected or diminution of the disease state is achieved.
  • the left common and external carotid arteries were isolated through a midline neck incision and ligated with a 4-0 silk suture (Ethicon).
  • An arteriotomy was fashioned in the common carotid artery just proximal to the carotid bifurcation.
  • a 4-0 nylon monofilament (Ethicon) was introduced through this incision into the internal carotid artery and advanced distal to the carotid bifurcation to occlude the origin of the MCA; the thread was carefully withdrawn 2 hours later.
  • tPA tissue plasminogen activator
  • Infarct volume Measuring infarct volume. The brain was removed and sectioned coronally into 2-mm segments. Brain slices were immersed in 2% 2,3,5-triphenyltetrazoliurn chloride (TTC) in saline, incubated for 30 minutes at 37°C and placed in 4% formalin/PBS overnight. The area of infarction remains unstained, appearing white, making it clearly distinguishable from stained viable tissue. The sections were photographed and the infarct area measured using the NIH computer image analysis program. Infarct volume was defined as the sum of the unstained areas of all sections, multiplied by their thickness and expressed in cubic millimeters. Data are represented as mean ⁇ SE. Differences were analyzed by ANOVA followed by the t-test and the level of significance was corrected using a post-hoc analysis with the Bonferroni test. Statistical significance was set at P ⁇ 0.05.
  • Tissue plasminogen activator tPA
  • tPA Tissue plasminogen activator
  • FIG. 1 Tissue plasminogen activator
  • Co-injection of EEIIMD reduced infarct size associated with tPA treatment significantly (FIGs. 1 and 2) at every dose tested (P ⁇ 0.01).
  • EEIIMD itself had no effect on infarct size in the absence of tPA (FIG. 1).
  • the control peptide EEIIMR which does not bind to tPA (Zhang et al., J. Biol.
  • tPA intracranial hemorrhage
  • ICH intracranial hemorrhage
  • the frequency and severity of ICH correlates with infarct volume. Based on the capacity of EEIIMD to reduce tPA- induced infarct size, ICH in tPA and tPA-EEIIMD-treated animals was next examined.
  • Parenchymal hemorrhage defined as a visible homogeneous mass of blood within the tissue, was analyzed using two parameters: 1) the prevalence of hemorrhage defined as the number of rats within each group that developed at least one ICH; and 2) the total number of intracerebral hemorrhages found in each group which takes into account that some animals develop bleeding at more than one site.
  • ti/ 2 of tPA in the circulation of rats was determined as previously reported (Nasar et al., ibid).
  • 125 I- tPA (6 mg per kg) was injected i.v., alone or together with 1 mg per kg EEIIMD.
  • Blood samples (0.25 mL) were taken 1, 4, 8 and 20 minutes after the injection and the radioactivity was measured.
  • the Pharsight ® WinNonlinTM nonlinear modeling software was used to determine the ti /2 for tPA and tPA + EEIIMD. The mean Un of tPA ⁇ EEIIMD were then compared using the t-test.
  • plasma fibrinolytic activity in animals treated with tPA or tPA/EEIIMD was measured ex vivo using a radiolabeled plasma clot assay as previously reported (Higazi et al. Blood 105: 1021-1028, 2005). Briefly, purified human fibrinogen was radiolabeled with 125 I and resuspended to a specific activity of 60,000 cpm/mL in PBS containing 3 mg per mL fibrinogen.
  • Clots were formed by placing 0.4 mL soluble fibrinogen in 16-mm diameter tissue culture wells (Nunc, Roskilde, Denmark) before adding thrombin (0.2 U/mL final concentration).
  • Radiolabeled fibrin clots were overlaid with PBS containing 1 nM tPA ( ⁇ 2 ⁇ M EEIIMD) or 25 ⁇ L of plasma prepared from control animals or animals treated with either tPA (6 mg per kg) or tPA (6 mg per kg) + EEIIMD (1 mg per kg) for two hours at 37°C, and the release of radiolabeled soluble fibrin degradation products was measured.
  • EEIIMD did not decrease the fibrinolytic activity of tPA in vivo, consistent with previous findings (Zhang et al., ibid).
  • the present results therefore indicate that the salutary effect of EEIIMD in stroke caused by vascular occlusion (e.g. in situ thrombosis) is not due to either enhanced clearance of tPA or to inhibition of its proteolytic activity.
  • the suspension of microemboli used to induce stroke contained 1.4 x 10 6 microparticles with an average diameter of 3 ⁇ m.
  • tPA (6 mg per kg) or tPA (6 mg per kg) with EEIIMD (1 mg per kg) in normal saline were injected i.v. two hours after the catheter was withdrawn.
  • Control groups received a vehicle consisting of normal saline alone or EEIIMD (1 mg per kg) in saline. 50% of the dose was given as a bolus injection and the remainder was infused over 60 min.
  • the rats were returned to their cages after recovering from anesthesia. Twenty-four hours after the procedure, the rats were euthanized with an overdose of Nembutal and the infarct size was measured.
  • FIG. 5 injection of tPA decreased infarct volume by approximately 48% (FIG. 5), which contrasts with its deleterious effect after mechanical stroke (FIG. 1).
  • animals treated with tPA and EEIIMD had significantly (P ⁇ 0.01) smaller infarct sizes than those treated with tPA alone (FIG. 5).
  • Pigs provide important advantages in modeling human cerebral injury.
  • the overall shape, gyral pattern, and distribution of gray and white matter are similar in pigs and humans, the response of the pigs to hypoxia and ischemia parallels that observed in humans, cerebral blood flow and metabolism are similar and piglet myelination and brain electrical activity mature in a similar manner. Therefore, the effect EEIIMD on the neurotoxicity of endogenous tPA in a piglet model of FPI was used as follows:
  • Pigs (1-5 days old) of either sex were used in these experiments. All protocols were approved by the Animal Use and Care Committee of the University of Pennsylvania. Pigs were sedated with isoflurane (1-2 minimum alveolar concentration). Anesthesia was obtained by i.v. injection of ⁇ -chloralose (30-50 mg per kg), supplemented as needed with i.v. injection of 5-10 mg per kg per h. A tracheotomy was performed. The animals were ventilated with room air and catheters were placed in femoral arteries and veins to monitor arterial blood pressure and to measure blood gases and pH. A cranial window was implanted to visualize pial arteries. A 1 cm hole was made in the skull contralateral to the cranial window site.
  • the dura was not removed at this site.
  • the opening was connected to a right angle metal shaft.
  • the metal shaft was sealed in the skull with dental acrylic.
  • the metal shaft was connected to a transducer housing and this in turn was connected to the fluid percussion device.
  • One end of the device was connected to the transducer housing while the other end had a Plexiglas piston mounted 0-ring.
  • the exposed end of this piston was covered with a rubber pad.
  • the entire system was filled with 0.9% saline (37°C).
  • Brain injury was caused by striking the piston with a 4.8 kg pendulum.
  • the pressure pulse was recorded on a storage oscilloscope, which was triggered photoelectrically by a sensor activated by the descent of the pendulum.
  • the amplitude of the pressure pulse was used as an index of the intensity of the injury.
  • the intensity of the insult was 1.9 ⁇ 0.1 atm at a constant duration of 19-23 m. sec.
  • FPI EEIIMD (1 mg per kg i.v.)-treated. Sham control animals underwent identical surgery as the FPI animals, but did not undergo brain injury. In FPI animals, vehicle or EEIIMD was administered 30 min prior to FPI. Animals were sacrificed for histopathology 4 hours after the trauma. Histologic preparation. Brains were perfused with heparinized saline, followed by 4% paraformaldehyde and then phosphate buffered saline. Coronal sections were prepared at 0.5 cm intervals for gross examination and photography.
  • staining was done on both frozen and paraffin embedded slides, blocks from some animals were cryoprotected in sucrose and serial sections were cut at 40 ⁇ m intervals from the front face of each frozen block and mounted on microscope slides. Sections (10 ⁇ m) were stained with hematoxylin and eosin and FluoroJade. Prior to staining, paraffin embedded slides were immersed in xylene/alcohol and rinsed in distilled water. The slides were then immersed in 0.06% potassium permanganate for 20 minutes on a rotating platform. The sections were rinsed in distilled water and transferred to a Fluorojade solution (0.001%) for 30 minutes on a rotating platform.
  • FIG. 6A shows hematoxilin and eosin (H & E) staining and FIG. 6B shows
  • degenerating neurons were also found in the hippocampal CA3 subfields of these animals (FIG. 6A, bottom left panels). In the brains of injured, FPI + EEIIMD- treated animals few swollen and degenerating neurons were found in the parietal cortex and hippocampal CA3 subfield (FIG. 6A, middle panels).
  • tPA/EEIIMD The effect of tPA/EEIIMD on brain swelling and brain edema after FPI was also examined.
  • H & E-stained coronal sections were used to evaluate tissue swelling by measuring the area of cortical tissue at the parietal lobe level of the brain. The parietal lobe was selected based on previous studies demonstrating that this region sustains the greatest cortical damage in this model.
  • brain edema was also assessed by measuring tissue water content.
  • tPA or isoproterenol prior to induction of brain injury decreased mean arterial blood pressure by 8 ⁇ 1 % and 9 ⁇ 1 % and increased pial artery diameter from 133 ⁇ 7 ⁇ m to 148 ⁇ 10 ⁇ m and 130 ⁇ 7 ⁇ m to 145 ⁇ 8 ⁇ m, respectively (P ⁇ 0.05).
  • Isoproterenol like tPA, significantly increased brain water content post-FPI, but unlike tPA the effect of isoproterenol was not inhibited by EEIIMD (FIG. 9). In the absence of FPI, isoproterenol had no effect on brain water content (FIG.
  • EEIIMD blood brain barrier permeability induced by traumatic brain injury (TBI).
  • TBI traumatic brain injury
  • EEIIMD protects against cerebral injury caused by both exogenous and endogenous tPA without affecting tPA clearance or fibrinolytic activity in vivo.
  • EEIIMD was shown to reduce infarct size and the incidence of ICH caused by transient mechanical occlusion of the MCA and to reduce infarct size caused by cerebral microemboli.
  • EEIIMD reduced edema formation and neuronal degeneration after traumatic brain injury.

Landscapes

  • 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

La présente invention concerne des utilisations thérapeutiques de peptides comprenant six résidus d'acides aminés dérivés des acides aminés 350-355 de l'inhibiteur 1 de l'activateur du plasminogène humain (PAI-1) pour prévenir un dommage neuronal dû en particulier à un accident cérébrovasculaire hypoxique ou thrombo-embolique et à une lésion cérébrale.
PCT/IL2007/001007 2006-08-10 2007-08-12 Peptides dérivés d'un inhibiteur de l'activateur du plasminogène pour prévenir un dommage neuronal Ceased WO2008018084A2 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US83666506P 2006-08-10 2006-08-10
US60/836,665 2006-08-10

Publications (2)

Publication Number Publication Date
WO2008018084A2 true WO2008018084A2 (fr) 2008-02-14
WO2008018084A3 WO2008018084A3 (fr) 2009-05-07

Family

ID=39033391

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/IL2007/001007 Ceased WO2008018084A2 (fr) 2006-08-10 2007-08-12 Peptides dérivés d'un inhibiteur de l'activateur du plasminogène pour prévenir un dommage neuronal

Country Status (1)

Country Link
WO (1) WO2008018084A2 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2009013753A1 (fr) * 2007-07-24 2009-01-29 Thrombotech Ltd. Peptides issus de l'inhibiteur 1 de l'activateur du plasminogène et leurs utilisations

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6750201B1 (en) * 1997-10-17 2004-06-15 The Trustees Of The University Of Pennsylvania Compositions and methods for promoting internalization and degradation of urokinase-type plasminogen activator

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2009013753A1 (fr) * 2007-07-24 2009-01-29 Thrombotech Ltd. Peptides issus de l'inhibiteur 1 de l'activateur du plasminogène et leurs utilisations
EP2468289A1 (fr) 2007-07-24 2012-06-27 Thrombotech Ltd. Peptides dérivés d'un inhibiteur-1 d'activateur de plasminigène et utilisations correspondantes
US8507436B2 (en) 2007-07-24 2013-08-13 D-Pharm Ltd. Peptides derived from plasminogen activator inhibitor-1 and uses thereof

Also Published As

Publication number Publication date
WO2008018084A3 (fr) 2009-05-07

Similar Documents

Publication Publication Date Title
AU2018205186B2 (en) Angiotensin in treating brain conditions
JP2002502594A (ja) 神経保護ペプチドおよびその使用
CN101808655B (zh) 衍生自纤溶酶原激活物抑制剂-1的肽和其应用
CA2940751C (fr) Utilisations d'un peptide d'angiotensine (1-7) pour le traitement retarde d'un accident vasculaire cerebral
US6103698A (en) Dolastatin-15 derivatives in combination with taxanes
WO2008018084A2 (fr) Peptides dérivés d'un inhibiteur de l'activateur du plasminogène pour prévenir un dommage neuronal
JP2017501967A (ja) 新規なペプチド組成物
HK1145397B (en) Peptides derived from plasminogen activator inhibitor-1 and uses thereof
EP1443953A2 (fr) Detection et traitement de lesions intravasculaires
WO2016010844A1 (fr) Procédés et compositions pour le traitement de l'accident vasculo-cérébral
WO2011086557A1 (fr) Peptides destinés au traitement de maladies respiratoires
HK1016998A (en) Increasing blood-brain barrier permeability with permeabilizer peptide derivatives of bradykinin

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: 07790060

Country of ref document: EP

Kind code of ref document: A2

NENP Non-entry into the national phase

Ref country code: DE

NENP Non-entry into the national phase

Ref country code: RU

122 Ep: pct application non-entry in european phase

Ref document number: 07790060

Country of ref document: EP

Kind code of ref document: A2