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
  • C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
  • C07K14/4702—Regulators; Modulating activity
  • C07K14/4705—Regulators; Modulating activity stimulating, promoting or activating activity
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
  • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
  • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
  • A61K47/56—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
  • A61K47/59—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes
  • A61K47/60—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
• • 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
  • A61P35/00—Antineoplastic agents
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
  • A61P7/06—Antianaemics
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K38/00—Medicinal preparations containing peptides

Definitions

• EPO-R agonists Given the immense potential of EPO-R agonists, both for studies of the important biological activities mediated by this receptor and for treatment of disease, there remains a need for the identification of peptide EPO-R agonists of enhanced potency and activity.
• the present invention provides such compounds.
• 1- naphthylalanine is 1-nal or Np
• N-methylglycine also known as sarcosine
• MeG or Sc N-methylglycine
• acetylated glycine N-acetylglycine
• the phrase “pharmaceutically acceptable” refers to molecular entities and compositions that are "generally regarded as safe”, e.g., that are physiologically tolerable and do not typically produce an allergic or similar untoward reaction, such as gastric upset, dizziness and the like, when administered to a human.
• 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.
• carrier refers to a diluent, adjuvant, excipient, or vehicle with which the compound is administered.
• C 4 of the spacer is covalently bonded to C 3 of X; N 1 of the spacer is covalently attached via a carbamate or an amide linkage to an activated PEG moiety; and N 2 of the spacer is covalently attached via a carbamate or an amide linkage to an activated PEG moiety, where PEG has a molecular weight of about 10,000 to about 60,000 Daltons (the term "about” indicating that in preparations of PEG, some molecules will weigh more, some less, than the stated molecular weight).
• N 2 represents the nitrogen atom of lysine's ⁇ -amino group and N 1 represents the nitrogen atom of lysine's ⁇ -amino group.
• two amine functional groups of the linker molecule are protected with the same removable amine protecting group.
• the protected linker is coupled to a solid support via the linker's third functional group.
• the two protected functional groups of the linker molecule are simultaneously deprotected, and the two peptide chains simultaneously synthesized on the deprotected amines. Note that using this technique, the sequences of the peptide chains of the dimer will be identical, and the thiol-protecting groups for the cysteine residues are all the same.
• each protected amino acid is generally reacted in about a 3- fold excess using an appropriate carboxyl group activator such as 2-(lH-benzotriazol-l-yl)-l, 1,3,3 tetramethyluronium hexafluorophosphate (HBTU) or dicyclohexylcarbodimide (DCC) in solution, for example, in methylene chloride (CH 2 CI 2 ), N-methyl pyrrolidone, dimethyl formamide (DMF), or mixtures thereof.
• carboxyl group activator such as 2-(lH-benzotriazol-l-yl)-l, 1,3,3 tetramethyluronium hexafluorophosphate (HBTU) or dicyclohexylcarbodimide (DCC) in solution, for example, in methylene chloride (CH 2 CI 2 ), N-methyl pyrrolidone, dimethyl formamide (DMF), or mixtures thereof.
• the amino-terminus of the peptide can be capped with an ⁇ - substituted acetic acid, wherein the ⁇ -substituent is a leaving group, such as an ⁇ -haloacetic acid, for example, ⁇ -chloroacetic acid, ⁇ -bromoacetic acid, or ⁇ -iodoacetic acid.
• an ⁇ -haloacetic acid for example, ⁇ -chloroacetic acid, ⁇ -bromoacetic acid, or ⁇ -iodoacetic acid.
• EPO-R agonist activity assays In vitro functional assays In vitro competitive binding assays quantitate the ability of a test peptide to compete with
• the peptides of this invention will find use in the treatment of renal insufficiency and/or end-stage renal failure/dialysis; chronic kidney disease, anemia associated with AIDS; anemia associated with chronic inflammatory diseases (for example, rheumatoid arthritis and chronic bowel inflammation) and autoimmune disease; and for boosting the red blood count of a patient prior to surgery.
• diluents could include carbohydrates, especially mannitol, ⁇ -lactose, anhydrous lactose, cellulose, sucrose, modified dextrans and starch.
• Certain inorganic salts may be also be used as fillers including calcium triphosphate, magnesium carbonate and sodium chloride.
• Some commercially available diluents are Fast-Flo, Emdex, STA-Rx 1500, Emcompress and Avicell.
• compositions for rectal or vaginal administration are preferably suppositories which may contain, in addition to the active substance, excipients such as cocoa butter or a suppository wax.
• Compositions for nasal or sublingual administration are also prepared with standard excipients well known in the art.
• Contemplated for use in the practice of this invention are a wide range of mechanical devices designed for pulmonary delivery of therapeutic products, including but not limited to nebulizers, metered dose inhalers, and powder inhalers, all of which are familiar to those skilled in the art.
• Some specific examples of commercially available devices suitable for the practice of this invention are the Ultravent nebulizer (Mallinckrodt Inc., St. Louis, MO); the Acorn II nebulizer (Marquest Medical Products, Englewood, CO); the Ventolin metered dose inhaler (Glaxo Inc., Research Triangle Park, NC); and the Spinhaler powder inhaler (Fisons Corp., Bedford, MA).
• IMDM medium 40-50 mL IMDM medium is added gently to cells: the medium is added drop by drop along the side of the 5OmL tube for the first 1OmL, and then the remaining volume of medium is slowly dispensed along the side of the tube. The cells are then spun at 900rpm for 20 min, and the media removed carefully by gentle aspiration. The cells are resuspended in ImI of IMDM medium and the cell density per mL is counted on hemacytometer slide (lO ⁇ L aliquot of cell suspension on slide, and cell density is the average count X 10,000 cells/ml). The cells are then diluted in IMDM medium to a cell density of 15,000 cells/mL.
• mice Forty eight hours following sample injection, the mice are administered an intraperitoneal injection of 0.2 ml of Fe 59 (Dupont, NEN), for a dose of approximately 0.75 ⁇ Curies/mouse.
• Mouse body weights are determined 24hr after Fe 59 administration, and the mice are sacrificed 48hr after Fe 59 administration.
• Blood is collected from each animal by cardiac puncture and hematocrits are determined (heparin was used as the anticoagulant).
• Each blood sample (0.2 ml) is analyzed for Fe 59 incorporation using a Packard gamma counter.
• Non-responder mice i.e., those mice with radioactive incorporation less than the negative control group
• Mice that have hematocrit values less than 53% of the negative control group are also eliminated.
• mice are dosed with four weekly bolus intravenous injections of either EPO positive control, test peptide, or vehicle.
• a range of positive control and test peptide doses, expressed as mg/kg, are tested by varying the active compound concentration in the formulation. Volumes injected are 5ml/kg.
• the vehicle control group is comprised twelve animals, while 8 animals are in each of the remaining dose groups. Daily viability and weekly body weights are recorded.
• Step 4 Synthesis of PEG moiety comprising two linear PEG chains linked by Lysine mPEG2-Lysinol-NPC
• Example 10 Synthesis of EPO-R agonist peptide homodimers of peptide monomers having the amino acid sequence (AcG)GL YACHMGPIT(l-nal) VCQPLR(MeG)K (SEQ ID NO: 2)
• Proliferation of the resulting cell line, BaF3/Gal4/Elk/EPOR is dependent on EPO-R activation.
• the degree of cell proliferation is quantitated using MTT, where the signal in the MTT assay is proportional to the number of viable cells.
• the starved cells are then washed twice with Dulbecco's PBS (Gibco), and resuspended to a density of IxIO 6 cells/ml in DMEM/F12 supplemented with 10% FBS (no WEHI-3 supernatant). 50 ⁇ L aliquots (-50,000 cells) of the cell suspension are then plated, in triplicate, in 96 well assay plates.
• test EPO mimetic peptides 50 ⁇ L aliquots of dilution series of test EPO mimetic peptides, or 50 ⁇ L EPO (R & D Systems Inc., Minneapolis, MN) or AranespTM (darbepoeitin alpha, an ERO-R agonist commercially available from Amgen) in DMEM/F12 media supplemented with 10% FBS (no WEHI-3 supernatant I) are added to the 96 well assay plates (final well volume of lOO ⁇ L). For example, 12 different dilutions may be tested where the final concentration of test peptide (or control EPO peptide) ranges from 81OpM to 0.0045pM. The plated cells are then incubated for 48h at 37°C.
• Peptide I Since the primary amino acid sequence of Peptide I differs from that of recombinant Human EPO (rHuEPO), it is less likely to induce a cross-reactive immune response against endogenous EPO. Although very rare, a cross-reactive immune response can cause serious side effects associated with loss of potency for both recombinant human ESAs and endogenous EPO. In addition, since Peptide I is a synthetic peptide, its production avoids the potential risk of contamination of the drug with host cell material that may occur with recombinant protein products.
• the patients selected for the study will be those pre-dialysis patients aged 18-75 years with hemoglobin >9 g/dL and ⁇ 11 g/dL secondary to chronic kidney disease who have not had previous treatment with ESAs and who meet eligibility criteria will be enrolled.
• Each replacement patient will be assigned to the same treatment group as the withdrawn patient. Patients who terminate from study after day 22 will not be replaced.
• a patient's hemoglobin level reaches 14 g/dL, the patient should be phlebotomized if clinically indicated. If a patient's hemoglobin level reaches 16 g/dL (confirmed), the patient should be phlebotomized.
• An open-label, multi-center, sequential, dose finding study of the safety, pharmacodynamics, and pharmacokinetics of Peptide I injection administered intravenously for the maintenance treatment of anemia in chronic hemodialysis patients will be performed to determine the range of monthly intravenously administered Peptide I doses that maintains hemoglobin within 1.0 g/dL above or below baseline in hemodialysis patients whose hemoglobin values were stable on Epoetin alfa.
• This study will also evaluate the safety profile of up to 3 doses of Peptide I administered intravenously in hemodialysis patients; evaluate the pharmacokinetic profile of up to 3 doses Peptide I administered intravenously in hemodialysis patients (in a subset of study patients).
• K/DOQI Outcomes Quality Initiative
• enrollment into the current cohort may be stopped and dose escalation to the next cohort is allowed if there are: no safety concerns are identified by the MM; and, 6 or more patients are transfused after week 4 or have a confirmed hemoglobin increase of ⁇ 1 g/dL at week 6.
• Figure 6 depicts an increase in hemoglobin from baseline of > 1 g/dL at 6 weeks following the first dose of 0.5 mg/kg, 0.1 mg/kg, 0.15 mg/kg, and 0.2 mg/kg of Peptide I (SEQ ID NO: 2) was observed in 17%, 73%, 55%, and 47% of patients, respectively.
• Xaa is N-acetylglycine
• Xaa is 1-naphthylalanine

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• 2008
  • 2008-07-23 WO PCT/US2008/070943 patent/WO2009025958A1/en not_active Ceased
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  • 2008-07-23 WO PCT/US2008/070926 patent/WO2009025957A1/en not_active Ceased
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