Classifications

• • 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
  • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
  • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
  • A61P1/18—Drugs for disorders of the alimentary tract or the digestive system for pancreatic disorders, e.g. pancreatic enzymes
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P13/00—Drugs for disorders of the urinary system
  • A61P13/12—Drugs for disorders of the urinary system of the kidneys
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P27/00—Drugs for disorders of the senses
  • A61P27/02—Ophthalmic agents
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P3/00—Drugs for disorders of the metabolism
  • A61P3/04—Anorexiants; Antiobesity agents
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P3/00—Drugs for disorders of the metabolism
  • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
  • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
• • 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
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P9/00—Drugs for disorders of the cardiovascular system
• • 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
  • C07K14/52—Cytokines; Lymphokines; Interferons
  • C07K14/521—Chemokines
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
  • C07K7/04—Linear peptides containing only normal peptide links
  • C07K7/08—Linear peptides containing only normal peptide links having 12 to 20 amino acids
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K38/00—Medicinal preparations containing peptides

Definitions

• the present invention relates to novel polypeptides, to nucleic acids encoding the same and to compositions comprising the same, as well as to their uses in therapeutic methods.
• disorders of carbohydrate metabolism occur in many forms. The most common disorders are acquired. Acquired or secondary derangements in carbohydrate metabolism, such as diabetic ketoacidosis, hyperosmolar coma, and hypoglycemia, all affect the central nervous system. Many forms and variants of peripheral nerve disease also are seen in diabetes. The remaining disorders of carbohydrate metabolism are the rare inborn errors of metabolism (i.e. genetic defects).
• the acquired disorders of carbohydrate metabolism are fairly common, both in the United States and internationally.
• Hypoglycemia is a common cause of neurological disease, especially acute mental deterioration, memory loss, disorientation, obtundation, and coma, among both alcoholics and patients with diabetes who are treated with insulin.
• Hyperinsulinemia from other causes is rare, but pancreatic tumors could be the cause.
• Diabetes with its various neurological complications, is among the most common disorders treated in adult patients.
• Diabetes is the most common endocrine disease, and is characterized by abnormalities of glucose metabolism.
• the abnormal glucose metabolism associated with this disease results in hyperglycemia (high blood glucose levels) and eventually causes complications of multiple organ systems, including eyes, kidneys, nerves, and blood vessels.
• Patients with persistent hyperglycemia or abnormal glucose tolerance are generally diagnosed with the disease, although most commonly patients initially present with excessive urination (polyuria) and frequent drinking due to extreme thirst (polydipsia). These typical initial symptoms result from the osmotic effects of hyperglycemia.
• the pathogenesis of diabetes mellitus is typically associated with pancreatic dysfunction, particularly of the beta cells of the pancreatic islets of Langerhans. This dysfunction may lead to destruction of the islet beta cells, which produce insulin, a glucose regulatory peptide hormone. Diabetes mellitus has been generally categorized as insulin dependent or type I, versus non-insulin dependent, or type II.
• the principal three forms or diabetes are:
• Type I Results from the body's failure to produce insulin. Treatment usually involves insulin administration.
• Type II Results from a condition in which the body fails to use insulin properly, combined with relative insulin deficiency. Many people destined to develop type II diabetes spend many years in a state of Pre-diabetes, a condition that occurs when a person's blood glucose levels are higher than normal but not high enough for a diagnosis of type II diabetes.
• Gestational diabetes Pregnant women who have never had diabetes before but who have high blood sugar (glucose) levels during pregnancy are said to have gestational diabetes. Gestational diabetes affects about 4% of all pregnant women. It may precede development of type II (or rarely type I).
• diabetes Many other forms of diabetes are categorized separately from these. Examples include congenital diabetes due to genetic defects of insulin secretion, cystic fibrosis-related diabetes, steroid diabetes induced by high doses of glucocorticoids, and several forms of monogenic diabetes.
• I insulin dependent diabetes mellitus
• pre-IDDM refers to an autoimmune condition that can be detected by biopsy or by analysis of autoimmune responses, in which pancreatic islet beta cells are being subject to a specific autoimmune attack to an extent where some cells may be subject to destruction.
• pre-IDDM the destruction (if any) has not progressed to an extent sufficient to require the administration of insulin. Since there can be a point in the early stages of Type I diabetes in which overt symptoms are observed but some islet function remains (known as the "honeymoon period", not all Type I diabetes is classified as IDDM, and not all pre-IDDM presents without overt symptoms.
• the metabolic complications associated with the abnormal metabolism caused by insulin insufficiency can affect numerous organ systems.
• the most common acute metabolic complication is that of diabetic ketoacidosis, characterized by severe hyperglycemia (and resulting hypovolemia caused by osmotic diuresis) as well as metabolic acidosis induced by excess free fatty acid release and the production of ketone bodies.
• Diabetic retinopathy is a leading cause of blindness, and is initiated by increased permeability of retinal capillaries which can progress to occlusion, hemorrhage, aneurysm formation, and neovascularization known as proliferative retinopathy.
• an object of the present invention is to provide polypeptides having an improved activity in comparison to the one of PRT8, as well as nucleic acid, vectors, and cells encoding the same.
• Another object of the present invention is to provide methods and uses of sbPRT8 for therapeutic treatment of diseases or disorders associated with an excess of glucose in blood (i.e. hyperglycemia).
• an isolated polypeptide comprising (a) the amino acid sequence set forth in SEQ ID NO: 1; or (b) an amino acid sequence that is at least 95% identical to SEQ ID NO: 1, wherein said polypeptide is able to lower glucose level in blood.
• the isolated polypeptide consists of the amino acid sequence set forth in SEQ ID NO: 1.
• the polypeptide is modified at the N-terminal end with an acetyl moiety and at the C-terminal end with an amide moiety.
• the invention further provides a pharmaceutical composition
• a pharmaceutical composition comprising an isolated polypeptide as defined above and at least one pharmaceutically acceptable carrier, excipient, or diluent.
• the invention further provides a therapeutically effective amount of the polypeptide described above for use for in lowering blood glucose level in a subject.
• said subject suffers from a disease or disorder associated with hyperglycemia.
• disease or disorder may be selected, but is not limited to, diabetes mellitus type 1, diabetes mellitus type 2, metabolic syndrome, obesity, nephropathy, retinopathy, cardiovascular diseases, glands dysfunction, pancreas diseases, sepsis, intracranial diseases, and post- surgery stress.
• the invention still further provides a therapeutically effective amount of the polypeptide described above for use in lowering plasma triglycerides (TG) blood level in a subject.
• TG plasma triglycerides
• the invention further provides a therapeutically effective amount of the polypeptide as described above for use in lowering the level of glycated hemoglobin (HbAlc) in blood of a subject.
• HbAlc glycated hemoglobin
• the invention further provides isolated nucleic acid molecules encoding a polypeptide comprising (a) the amino acid sequence set forth in SEQ ID NO: 1; or (b) an amino acid sequence that is at least 95% identical to SEQ ID NO: 1, expression vectors comprising the same, and host cells comprising said expression vector.
• FIGS 1A-1B Effects of sbPRT8 on C57B1/6 mice treated with streptozotocyn (STZ)
• Figure 1A Graph showing glucose levels in C57B1/6 mice injected with STZ (IP injection of 50 mg/kg for 4 days) followed by a 5 weeks treatment with either PRT8 or sbPRT8, and IP injected with 1 mg/kg of glucose (IPGTT);
• Figure IB Graph showing glucose levels in C57B1/6 mice injected with STZ (IP injection of 50mg/kg for 4 days) followed by a 4 weeks treatment with either PRT8 or sbPRT8, and IP injected with 1 mg/kg of glucose (IPGTT);
• WO 2011/016026 disclosed novel proteins named PRT5, PRT6, PRT7 and PRT8.
• PRT8 was found particularly in the pancreas and testis, and also in the liver. In particular, it has been shown that PRT8 is able to significantly lower the glucose level in blood.
• a shortened PRT8 peptide namely sbPRT8
• sbPRT8 has an improved biological activity over the full length PRT8 peptide, and is able to lower blood glucose level, even more significantly.
• the present invention provides an isolated polypeptide comprising an amino acid sequence as set forth in SEQ ID NO: 1, as well as homologs and derivatives thereof, wherein said polypeptide lower glucose level in blood (i.e. decrease the glucose concentration in blood).
• polypeptide is used herein to denote a polypeptide or a protein.
• the polypeptide may be obtained synthetically, through genetic engineering methods, expression in a host cell, or through any other suitable means. Unless indicated otherwise, a polypeptide is generally composed of naturally- occurring L-amino acids.
• the polypeptide of the invention is modified at the C- terminal end and/or at the N-terminal end.
• the peptide is modified as follows: acetyl-(Peptide Sequence)-amide.
• the polypeptide of the invention is modified with one or more groups selected from a biotin group, a fluorescent group or a cysteine residue.
• modified polypeptide With respect to a modified polypeptide, fragments, homologues and derivatives thereof, in connection with the present invention, it is understood that it retains the biological activity of the polypeptide of SEQ ID NO: 1.
• one or more assays can be carried out, such as for example an in vitro, in vivo or a clinical experiment in which a modified polypeptide is compared to the corresponding unmodified one that is assayed in parallel or in a separately conducted experiment.
• a modified polypeptide or variant polypeptide may be a polypeptide that includes at least two, three or more copies of sbPRT8 optionally separated by an amino acid spacer.
• a modified polypeptide may be also a polypeptide having a degree of identity being at least 70%, preferably at least 80%, more preferably at least 90% and particularly at least 95% with SEQ ID NO:l.
• polypeptides derived from sbPRT8 e.g., modified polypeptides in which one or more amino acids are replaced by another amino acid by conservative substitution.
• conservative substitution refers to the substitution of an amino acid in one class by an amino acid of the same class, where a class is defined by common physicochemical amino acid side chain properties and high substitution frequencies in homologous proteins found in nature.
• Class I Cys
• Class II Ser, Thr, Pro, Ala, Gly
• Class III Asn, Asp, Gin, Glu
• Class IV His, Arg, Lys
• Class V He, Leu, Val, Met
• Class VI Phe, Tyr, Trp
• substitution of an Asp for another class III residue such as Asn, Gin, or Glu is a conservative substitution.
• substitutions are made in the amino acid sequence. In another embodiment, two substitutions are made. In a further embodiment, three substitutions are made. The maximum number of substitutions should not exceed the number of amino acids which leaves at least 70%, desirably at least 80%, preferably at least 90%, most preferably at least 95% of the amino acids in the unsubstituted sequence. In one particular embodiment, the substitutions which include up to 3, at times up to 6 amino acid residues substituted by others, are conservative substitutions. In a further embodiment, one or more amino acids may be replaced by D-amino acids, preferably the corresponding D-amino acids. In a particular embodiment, all of the amino acids are D-amino acids.
• substitutions are changes that would not be expected to alter the secondary structure of the polypeptide, i.e., conservative changes.
• the following list shows amino acids (right side) that may be exchanged for the original amino acids (left side).
• Val He; Leu Amino acids can also be grouped according to their essential features, such as charge, size of the side chain, and the like. The following list shows groups of similar amino acids. Preferred substitutions would exchange an amino acid present in one group with an amino acid from the same group, as follows:
• Polar negatively charged residues and their amides Asp, Asn, Glu, Gin; 3. Polar positively charged residues: His, Arg, Lys;
• the polypeptide of the invention can be produced by conventional chemical methods, such as solid phase synthesis (using e.g. FMOC and BOC techniques), and solution phase synthesis. These proteins or polypeptides may also be produced in bacterial or insect cells or other eukaryotic transcriptional in vivo system. Following production, the polypeptide are purified from the cells in which they have been produced. Polypeptide purification and isolation methods are known to the person of skill in the art.
• the polypeptide may be produced as a fusion with a second protein, such as Glutathione-S-transferase (GST) or the like, or a sequence tag, such as the Histidine tag (His-tag) sequence.
• GST Glutathione-S-transferase
• His-tag Histidine tag
• the polypeptide of the invention can also be synthesized in cell-free systems, using, for example, cell extracts or ribosomes.
• the polypeptide of the invention may be further modified to improve their function, affinity, or stability.
• cyclization may be used to impart greater stability and/or overall improved performance upon a polypeptide.
• side chain cyclization and backbone cyclization have been developed, including side chain cyclization and backbone cyclization. These methods are well documented in the prior art.
• a particular method of cyclization involves stabilization of an amphipathic alpha-helix by using para-substituted amino acid derivatives of a benzene ring.
• Another particular method of cyclization is backbone cyclization.
• Another method of cyclization which involves backbone- to side chain connections may also be used.
• polypeptide of the invention may be extended at the N-terminus and/or C-terminus thereof with various identical or different organic moieties which are not naturally occurring or synthetic amino acids.
• polypeptide may be extended at the N-terminus and/or C-terminus thereof with an N-acetyl group.
• the polypeptide of the invention can be coupled through their N-terminus to a lauryl-cysteine (LC) residue and/or through their C-terminus to a cysteine (C) residue, or to other residue/s suitable for linking the polypeptide to adjuvant/s for immunization.
• LC lauryl-cysteine
• C cysteine
• the present invention provides an isolated nucleic acid encoding a polypeptide as set forth in SEQ ID NO: 1 and any derivatives and analogs thereof.
• nucleic acid molecule is intended to include DNA molecules (e.g., cDNA) and RNA molecules (e.g., mRNA) and analogs of the DNA or RNA generated using nucleotide analogs.
• the nucleic acid molecule can be single-stranded or double- stranded, but preferably it is double- stranded DNA.
• isolated nucleic acid molecule is intended to include nucleic acid molecules which are separated from other nucleic acid molecules and which are substantially free of other cellular material, or culture medium when produced by recombinant techniques, or substantially free of chemical precursors or other chemicals when chemically synthesized.
• Polynucleotide or nucleic acid derivatives differ from the sequences described or known in nucleotide sequence.
• a polynucleotide derivative may be characterized by one or more nucleotide substitutions, insertions, or deletions.
• a nucleic acid molecule of the present invention i.e. a nucleic acid molecule having a nucleotide sequence encoding sbPRT8, can be generated using standard molecular biology techniques and the sequence information provided herein.
• the present invention provides a vector comprising an isolated nucleic acid molecule encoding the polypeptide as set forth in SEQ ID NO: 1 or any functional derivative or analog thereof.
• said nucleic acid molecule is operably linked to a promoter.
• said vector is an expression vector.
• the term "vector" is intended to include a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked.
• a vector may be characterized by one or a small number of restriction endonuclease sites at which such DNA sequences may be cut in a determinable fashion without the loss of an essential biological function of the vector, and into which a DNA fragment may be spliced in order to bring about its replication and cloning.
• a vector may further contain a marker suitable for use in the identification of cells transformed with the vector.
• plasmid which refers to a circular double stranded DNA loop into which additional DNA segments can be ligated.
• Another type of vector is a viral vector, wherein additional DNA segments can be ligated into the viral genome.
• vectors are capable of autonomous replication in a host cell into which they are introduced (e.g., bacterial vectors having a bacterial origin of replication and episomal mammalian vectors).
• Other vectors e.g., non-episomal mammalian vectors
• certain vectors are capable of directing the expression of genes to which they are operatively linked. Such vectors are referred to herein as "expression vectors”.
• expression vectors of utility in recombinant DNA techniques are often in the form of plasmids.
• plasmid and "vector” can be used interchangeably as the plasmid is the most commonly used form of vector.
• the invention is intended to include such other forms of expression vectors, such as viral vectors (e.g., replication defective retroviruses, adenoviruses and adeno-associated viruses), which serve equivalent functions.
• viral vectors e.g., replication defective retroviruses, adenoviruses and adeno-associated viruses
• operatively linked or “operably linked” is intended to mean that molecules are functionally coupled to each other in that the change of activity or state of one molecule is affected by the activity or state of the other molecule.
• Nucleotide sequences are "operably linked" when the regulatory sequence functionally relates to the DNA sequence encoding the polypeptide or protein of interest.
• a promoter nucleotide sequence is operably linked to a DNA sequence encoding the polypeptide of interest if the promoter nucleotide sequence controls the transcription of the DNA sequence encoding the protein of interest.
• two polypeptides that are operatively linked are covalently attached through peptide bonds.
• the present invention provides a cell comprising the vector as described above, said vector comprising an isolated nucleic acid molecule encoding the polypeptide as set forth in SEQ ID NO: 1 and any derivatives or analog thereof.
• said cell is a host cell selected from the group consisting of: a plant cell, an insect cell, a fungal cell, a bacterial cell or a mammalian cell.
• the terms "host cell” and “recombinant host cell” are used interchangeably herein.
• a “host cell” includes any cultivatable cell that can be modified by the introduction of heterologous DNA.
• a host cell is one in which a transcriptional regulatory protein can be stably expressed, post- translationally modified, localized to the appropriate sub-cellular compartment, and made to engage the appropriate transcription machinery.
• the choice of an appropriate host cell will also be influenced by the choice of detection signal.
• reporter constructs as described above, can provide a selectable or screenable trait upon activation or inhibition of gene transcription in response to a transcriptional regulatory protein; in order to achieve optimal selection or screening, the host cell phenotype will be considered. It is understood that such terms refer not only to the particular subject cell but to the progeny or potential progeny of such a cell. Because certain modifications may occur in succeeding generations due to either mutation or environmental influences, such progeny may not, in fact, be identical to the parent cell, but are still included within the scope of the term as used herein.
• a host cell of the present invention includes prokaryotic cells and eukaryotic cells.
• Prokaryotes include gram negative or gram positive organisms, for example, E. coli or Bacilli.
• Suitable prokaryotic host cells for transformation include, for example, E. coli, Bacillus subtilis, Salmonella typhimurium, and various other species within the genera Pseudomonas, Streptomyces, and Staphylococcus.
• Eukaryotic cells include, but are not limited to, yeast cells, plant cells, fungal cells, insect cells (e.g., baculovirus), mammalian cells, and the cells of parasitic organisms, e.g., trypanosomes.
• yeast includes not only yeasts in a strict taxonomic sense, i.e., unicellular organisms, but also yeast-like multicellular fungi of filamentous fungi.
• Exemplary species include Kluyverei lactis, Schizosaccharomyces pombe, and Ustilaqo maydis, with Saccharomyces cerevisiae being preferred.
• Other yeast which can be used in practicing the present invention are Neurospora crassa, Aspergillus niger, Aspergillus nidulans, Pichia pastoris, Candida tropicalis, and Hansenula polymorpha.
• Mammalian host cell culture systems include established cell lines such as HeLa cells, COS cells, L cells, 3T3 cells, Chinese hamster ovary (CHO) cells, embryonic stem cells, etc.
• the present invention provides a composition comprising an isolated polypeptide sbPRT8 wherein said polypeptide comprises the sequence as set forth in SEQ ID NO: 1, or any analogs of derivatives thereof, as defined above.
• any composition provided in the present invention may further comprise a pharmaceutically acceptable adjuvant, carrier, excipient, or diluent.
• the term “pharmaceutically acceptable carrier” it is meant any one of inert, non-toxic materials, which do not react with the active ingredient.
• the carrier is selected at times based on the desired form of the formulation.
• the carrier may also at times have the effect of the improving the delivery or penetration of the active ingredient to the target tissue, for improving the stability of the drug, for slowing clearance rates, for imparting slow release properties, for reducing undesired side effects etc.
• the carrier may also be a substance that stabilizes the formulation (e.g. a preservative), for providing the formulation with an edible flavor, etc.
• the carriers may be any of those conventionally used and is limited only by chemico-physical considerations, such as solubility and lack of reactivity with the antibodies of the invention, and by the route of administration.
• the carrier may include additives, colorants, diluents, buffering agents, disintegrating agents, moistening agents, preservatives, flavoring agents, and pharmacologically compatible carriers.
• the carrier may be an adjuvant, which, by definition are substances affecting the action of the active ingredient in a predictable way.
• Typical examples of carriers include (a) liquid solutions, where an effective amount of the active substance is dissolved in diluents, such as water, saline, natural juices, alcohols, syrups, etc.; (b) capsules (e.g.
• the ordinary hard- or soft-shelled gelatin type containing, for example, surfactants, lubricants, and inert fillers), tablets, lozenges (wherein the active substance is in a flavor, such as sucrose and acacia or tragacanth or the active substance is in an inert base, such as gelatin and glycerin), and troches, each containing a predetermined amount of active agent as solids or granules; (c) powders; (d) suspensions in an appropriate liquid; (e) suitable emulsions; (f) liposome formulation; and others.
• compositions of the invention may also optionally further comprise additional active agents, such as, but not limited to antibiotics, cytokines, lymphokines, growth factors, hormones, anti-oxidants, vitamins, etc.
• additional active agents such as, but not limited to antibiotics, cytokines, lymphokines, growth factors, hormones, anti-oxidants, vitamins, etc.
• the present invention provides the use of a polypeptide sbPRT8 for the preparation of a medicament for the treatment of a disease or disorder associated with hyperglycemia, in particular a disease or disorder selected from the group consisting of diabetes mellitus type 1, diabetes mellitus type 2, metabolic syndrome, obesity, nephropathy, retinopathy, cardiovascular diseases, glands dysfunction, pancreas diseases, sepsis, intracranial diseases, and post-surgery stress.
• the term "effective amount” means an amount necessary to achieve a selected result, which at present, involves the amount of sbPRT8, or biologically active derivatives thereof, necessary for treating a disorder.
• Said therapeutic effective amount, or dosing is dependent on severity and responsiveness of the disease state to be treated, with the course of treatment lasting one hour to several hours, one day to several days, or until a cure is effected or a diminution of the disease state is achieved.
• Persons of ordinary skill can readily determine optimum dosages, dosing methodologies and repetition rates.
• Optimum dosages may vary depending on the relative potency of each polypeptide or protein of the invention, or compositions comprising thereof, and can generally be estimated based on EC50, found to be effective in in vitro as well as in in vivo animal models. Persons of ordinary skill in the art can easily estimate repetition rates for dosing based on measured residence times, concentrations, and adjustment to the employed polypeptide or protein.
• treat, treating or treatment mean ameliorating one or more clinical indicia of disease activity in a patient having a disease or disorder. “Treatment” refers to therapeutic treatment.
• patient or “subject in need” is meant any mammal for which treatment of a disorder or disease is desired in order to overcome said disorder or disease, particularly a human subject.
• a “therapeutically effective amount” is also determined by the severity of the disease in conjunction with the preventive or therapeutic objectives, the route of administration and the patient's general condition (age, sex, weight and other considerations known to the attending physician).
• Various methods of administration may be used for delivering sbPRT8 described in the invention to a subject in need.
• Said polypeptide, or compositions comprising the same may be delivered via intravenous (i.v.), intramuscular (i.m.), intraperitoneal (i.p.), or topical injections, or through any other route found suitable by the man skilled in the art.
• the polypeptides or proteins of the invention should be prepared in a way that would enable their stability in the system following administration.
• disorder refers to a condition in which there is a disturbance of normal functioning.
• a “disease” is any abnormal condition of the body or mind that causes discomfort, dysfunction, or distress to the person affected or those in contact with the person.
• the term is used broadly to include injuries, congenital malformations, disabilities, syndromes, symptoms, deviant behaviors, and atypical variations of structure and function, chronic or permanent health defects resulting from disease.
• said subject may be a mammal, human or non-human.
• Non-human mammals include, but are not limited to cows, horses, dogs, cats, mice, rats, guinea-pigs, etc.
• the subject is a human, particularly a patient, or a healthy individual.
• a "disease or disorder associated with hyperglycemia” refers to diseases or disorders in which an excessive amount of glucose circulates in the blood plasma. This is generally a glucose level higher than 11.1 mmol/1 (200 mg/dl), but symptoms may not start to become noticeable until even higher values such as 15-20 mmol/1 ( ⁇ 250-300 mg/dl). According to the American Diabetes Association guidelines, a subject with a consistent range between 100 and 126 is considered hyperglycemic, while above 126 mg/dl or 7 mmol/1 is generally held to have diabetes. Chronic levels exceeding 7 mmol/1 (125 mg/dl) can produce organ damage.
• Diabetes mellitus type 1 and type 2
• cardiovascular diseases e.g. stroke, myocardial infarction
• glands dysfunction e.g. thyroid, adrenal, or pituitary
• pancreas diseases e.g. sepsis
• intracranial diseases e.g encephalitis, brain tumors, brain bleeds, meningitis.
• Prolonged, major surgeries can temporarily increase glucose levels.
• acute hyperglycemia increases plasma triglycerides (TG) by stimulating hepatic TG secretion, in a manner which is independent of either plasma insulin or free fatty acids levels.
• TG plasma triglycerides
• glycated hemoglobin (also referred to as hemoglobin Ale, HbAlc, AlC, Hblc, or HbAlc) is a form of hemoglobin that is measured primarily to identify the average plasma glucose concentration over prolonged periods of time. It is formed in a non-enzymatic glycation pathway by hemoglobin's exposure to plasma glucose. Normal levels of glucose produce a normal amount of glycated hemoglobin. As the average amount of plasma glucose increases (hyperglycemia), the fraction of glycated hemoglobin increases in a predictable way. In diabetes mellitus, higher amounts of glycated hemoglobin have been associated with cardiovascular disease, nephropathy, and retinopathy.
• STZ streptozocin
• mice C57B1/6 males mice were purchased from Harlan Laboratories Ltd., Jerusalem, Israel, After 7 days of rest, mice were IP injected with 50 mg/kg STZ for 4 days. Five days after STZ injection, mice were treated as follows:
• mice were starved for food only for 12h and then were IP injected with lmg/kg of glucose solution (IPGTT).
• IPGTT glucose solution
• mice C57B1/6 males mice were purchased from Harlan Laboratories Ltd., Jerusalem, Israel. After 7 days of rest, mice were IP injected with 50 mg/kg STZ for 4 days. Five days after STZ injection, mice were treated as follows:
• mice were starved for food only for 12h and then were IP injected with 1 mg/kg of glucose solution (IPGTT).
• IPGTT insulin solution

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• 2014
  • 2014-12-30 AU AU2014374945A patent/AU2014374945A1/en not_active Abandoned
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  • 2014-12-30 WO PCT/IL2014/051136 patent/WO2015101984A1/en active Application Filing
  • 2014-12-30 CN CN201480072200.6A patent/CN105916874A/zh active Pending
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  • 2014-12-30 CA CA2935624A patent/CA2935624A1/en not_active Abandoned
• 2016
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