COMPOSITIONS AND METHODS FOR IMPROVING PRODUCTION OF RECOMBINANT POLYPEPTIDES

• US 20100260706A1
• Filed: 12/23/2009
• Published: 10/14/2010
• Est. Priority Date: 08/15/2007
• Status: Abandoned Application

• Current Assignee: Amunix Operating Inc. (Sanofi )
• Original Assignee: Amunix Operating Inc. (Sanofi )
• Inventors: Yin, Yong, Bogin, Oren, Geething, Nathan C., Schellenberger, Volker, Wang, Chia-wei, Stemmer, Willem P.
• Application Number:
  US12/646,566
• Publication Number:
  US 20100260706A1
• Time in Patent Office: Days
• US Class Current: 424/85.200
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61P 1/02 Stomatological preparations, e.g. drugs for caries, aphtae, periodontitis
• CPC Class Codes: A61P 1/04 for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 11/06 Antiasthmatics
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 15/00 Drugs for genital or sexual disorders for disorders of sex hormones A61P5/24; Contraceptives
• CPC Class Codes: A61P 15/10 for impotence
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 17/06 Antipsoriatics
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 19/10 for osteoporosis
• CPC Class Codes: A61P 21/04 for myasthenia gravis
• CPC Class Codes: A61P 25/04 Centrally acting analgesics, e.g. opioids
• CPC Class Codes: A61P 25/14 for treating abnormal movements, e.g. chorea, dyskinesia
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 27/06 Antiglaucoma agents or miotics
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 3/04 Anorexiants; Antiobesity agents
• CPC Class Codes: A61P 3/10 : for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 31/00 : Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 31/04 : Antibacterial agents
• CPC Class Codes: A61P 31/12 : Antivirals
• CPC Class Codes: A61P 31/18 : for HIV
• CPC Class Codes: A61P 35/00 : Antineoplastic agents
• CPC Class Codes: A61P 35/02 : specific for leukemia
• CPC Class Codes: A61P 37/08 : Antiallergic agents antiasthmatic agents A61P11/06; ophthalmic antiallergics A61P27/14
• CPC Class Codes: A61P 43/00 : Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 5/00 : Drugs for disorders of the endocrine system
• CPC Class Codes: A61P 5/24 : of the sex hormones
• CPC Class Codes: A61P 7/00 : Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: A61P 7/02 : Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
• CPC Class Codes: A61P 7/06 : Antianaemics
• CPC Class Codes: A61P 9/00 : Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/10 : for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: A61P 9/12 : Antihypertensives
• CPC Class Codes: C07K 14/00 : Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
• CPC Class Codes: C07K 14/535 : Granulocyte CSF; Granulocyte-macrophage CSF
• CPC Class Codes: C07K 14/56 : IFN-alpha
• CPC Class Codes: C07K 14/61 : Growth hormone [GH], i.e. somatotropin
• CPC Class Codes: C07K 16/2863 : against receptors for growth factors, growth regulators
• CPC Class Codes: C07K 16/30 : from tumour cells
• CPC Class Codes: C07K 16/32 : against translation products of oncogenes
• CPC Class Codes: C07K 16/40 : against enzymes
• CPC Class Codes: C07K 2317/22 : from camelids, e.g. camel, llama or dromedary
• CPC Class Codes: C07K 2317/55 : Fab or Fab'
• CPC Class Codes: C07K 2317/569 : Single domain, e.g. dAb, sdAb, VHH, VNAR or nanobody®
• CPC Class Codes: C07K 2317/622 : Single chain antibody (scFv)
• CPC Class Codes: C07K 2317/626 : Diabody or triabody
• CPC Class Codes: C07K 2319/00 : Fusion polypeptide
• CPC Class Codes: C07K 2319/33 : fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies

Abstract

The present invention relates to biologically active polypeptides linked to one or more accessory polypeptides. The present invention also provides recombinant polypeptides including vectors encoding the subject proteinaceous entities, as well as host cells comprising the vectors. The subject compositions have a variety of utilities including a range of pharmaceutical applications.

Claims

1-148. -148. (canceled)

149. A biologically active protein comprising at least two domains wherein (a) a first domain of said at least two domains comprises an amino acid sequence having and/or mediating said biological activity;

and (b) a second domain of said at least two domains comprises an amino acid sequence consisting preferably of at least about 100 amino acid residues forming random coil conformation whereby said random coil conformation mediates an increased in vivo and/or in vitro stability of said biologically active protein.

150. The biologically active protein according to claim 149, wherein said second domain forming random coil conformation consists of alanine, serine and proline residues.

151. The biologically active protein according to claim 149 or 150, wherein said second domain forming random coil conformation comprises a plurality of amino acid repeats, wherein said repeat consist of Ala, Ser, and Pro residues and wherein no more than 6 consecutive amino acid residues are identical.

152. The biologically active protein according to any one of claims 149 to 151, wherein said proline residues constitute more than 4% and less than 40% of the amino acids of said second domain forming random coil conformation.

153. The biologically active protein according to any one of claims 149 to 152, wherein said second domain of said at least two domains comprises an amino acid sequence consisting of about 100 to 3000 amino acid residues forming random coil conformation.

154. The biologically active protein according to any one of claims 149 to 153, wherein said polypeptide with biological activity is selected from the group consisting of binding molecules, antibody fragments, cytokines, growth factors, hormones or enzymes.

155. The biologically active protein according to claim 154 wherein said binding molecule is selected from the group consisting of antibodies, Fab fragments, F(ab′

)2 fragments, CDR derived peptidomimetics, single chain variable fragments (scFv), domain antibodies and lipocalins.

156. The biologically active protein according to any one of claims 149 to 154, wherein said polypeptide with biological activity is selected from the group consisting of granulocyte colony stimulating factor, human growth hormone, alphainterferon, betainterferon, gammainterferon, tumor necrosis factor, erythropoietin, coagulation factor VIII, gp120/gp160, soluble tumor necrosis factor I and II receptor, interleukin 2 and neutrophil gelatinase associated lipocalin.

157. The biologically active protein according to any one of claims 149 to 156, wherein said increased in vivo stability of said biologically active protein is a prolonged plasma half life of said biologically active protein comprising said random coil forming second domain when compared to said biologically active protein lacking said random coil forming second domain.

158. A composition comprising the biologically active protein according to any one of claims 149 to 157.

159. The composition according to claim 158, which is a pharmaceutical composition, optionally further comprising a pharmaceutical acceptable carrier.

160. A nucleic acid molecule encoding the biologically active protein of any one of claims 149 to 157.

161. A vector comprising the nucleic acid of claim 159.

162. A cell comprising the nucleic acid according to claim 160 or the vector according to claim 161.

163. A method for the preparing a biologically active protein comprising culturing the cell according to claim 162 and isolating said biologically active protein from the culture.

164. A method of treating a disease condition selected from the group consisting of hormone deficiency related disorders, autoimmune disease, cancer, anaemia, neovascular diseases, infectious/inflammatory diseases, thrombosis, myocardial infarction, diabetes, reperfusion injury, and a kidney disease, comprising administering to a subject in need thereof a composition according to claim 159.