Classifications

• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K38/00—Medicinal preparations containing peptides
  • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
  • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
  • A61K38/22—Hormones
  • A61K38/24—Follicle-stimulating hormone [FSH]; Chorionic gonadotropins, e.g. HCG; Luteinising hormone [LH]; Thyroid-stimulating hormone [TSH]
• • 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
  • A61P3/06—Antihyperlipidemics
• • 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
• • 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
  • A61P5/00—Drugs for disorders of the endocrine system
  • A61P5/48—Drugs for disorders of the endocrine system of the pancreatic hormones
  • A61P5/50—Drugs for disorders of the endocrine system of the pancreatic hormones for increasing or potentiating the activity of insulin

Definitions

• TSH thyrotropin-stimulating hormone
• the TSH receptor (TSHR) is a member of the G-protein coupled, seven transmembrane receptor superfamily. Activation of the TSH receptor leads to coupling with heterotrimeric G proteins, which evoke downstream cellular effects.
• the TSH receptor has been shown to interact with G proteins of subtypes G s , G q , G 12 , and Gj. In particular, interaction with G s leads to activation of adenyl cyclase and increased levels of cAMP. See Laugwitz, K. L., et al.
• 40 ⁇ l of conditioned medium were collected for glycerol assay analysis, and 40 ⁇ l of conditioned medium were collected for free fatty acid analysis.
• the glycerol standard was diluted in water to a range from 200 nmols/10 ⁇ l to 0.25 nmols/10 ⁇ l.
• Glycerol was used as a reference for determining the amount of glycerol in the conditioned media.
• Sigma reagent A was reconstituted to the recommended concentration.
• Conditioned media samples were assayed in 96-well plates. 150 ⁇ l of Sigma reagent A were added to 10 ⁇ l of glycerol standard plus 40 ⁇ l of assay medium.
• the CHO culture supernatant was 0.2 ⁇ xn filtered and adjusted to pH 6 and 20 mM 2-Morpholinoethanesulfonic Acid (MES).
• MES 2-Morpholinoethanesulfonic Acid
• the CGH in the adjusted supernatant was captured at 55 cm/hr using a 1:2 online dilution with 20 mM MES pH 6 onto a POROS HS 50 column that was previously equilibrated in 20 mM MES pH 6. After loading was complete, the column was washed with 20 column volumes (CV) of equilibration buffer. This was followed by a 3 CV wash with 250 mM NaCl in 20 mM MES pH 6 at 90 cm/hr.
• CV column volumes
• ConA Sepharose Chromatography is Concanavalin A coupled to Sepharose.
• Concanavalin A is a lectin, which binds reversibly to molecules, which contain D-mannopyranosyl, D- glucopyranosyl and related residues.
• the adjusted pool of CGH from the cation exchange chromatography was applied directly at 2 cm/hr to the ConA column equilibrated in 20 mM Tris pH 7.4 containing 0.5 M NaCl. After loading, the column was washed with 20 CV of equilibration buffer.
• the CGH was then competed off the column at 1-2 cm/hr with 3 CV of 0.5M Methyl-D-Manno-Pyranoside in 20 mM Tris pH 7.4. This CGH pool was concentrated via ultrafiltration using an Amicon stirred cell with a 5kDa-cutoff membrane.
• the concentrated CGH ConA pool was then applied to an appropriately sized bed of Superdex 75 resin (i.e. ⁇ 5% of bed volume) for removal of remaining HMW contaminants and for buffer exchange into PBS.
• the CGH eluted from the Superdex 75 column at about 0.65 to 0.7 CV and was concentrated for storage at -80 °C using the Amicon stirred cell with a 5kDa-cutoff ultrafiltration membrane.
• the heterodimeric protein was pure by Coomassie-stained SDS PAGE, had the correct NH2 termini, the correct amino acid composition, and the correct mass by SEC MALS.
• the overall process recovery estimated by RP HPLC assay was 50-60%.

Landscapes

• Health & Medical Sciences (AREA)
• Life Sciences & Earth Sciences (AREA)
• Diabetes (AREA)
• Chemical & Material Sciences (AREA)
• General Health & Medical Sciences (AREA)
• Endocrinology (AREA)
• Veterinary Medicine (AREA)
• Engineering & Computer Science (AREA)
• Public Health (AREA)
• Bioinformatics & Cheminformatics (AREA)
• Medicinal Chemistry (AREA)
• Animal Behavior & Ethology (AREA)
• Pharmacology & Pharmacy (AREA)
• Organic Chemistry (AREA)
• Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
• General Chemical & Material Sciences (AREA)
• Chemical Kinetics & Catalysis (AREA)
• Hematology (AREA)
• Obesity (AREA)
• Emergency Medicine (AREA)
• Reproductive Health (AREA)
• Zoology (AREA)
• Gastroenterology & Hepatology (AREA)
• Immunology (AREA)
• Proteomics, Peptides & Aminoacids (AREA)
• Epidemiology (AREA)
• Child & Adolescent Psychology (AREA)
• Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Priority Applications (4)

Applications Claiming Priority (2)

Publications (1)

Family

ID=23180168

Family Applications (1)

Country Status (7)

Cited By (3)

Families Citing this family (4)

Citations (3)

Family Cites Families (2)

• 2002
  • 2002-07-15 CA CA002454181A patent/CA2454181A1/en not_active Abandoned
  • 2002-07-15 IL IL15952602A patent/IL159526A0/xx unknown
  • 2002-07-15 WO PCT/US2002/022747 patent/WO2003006051A1/en not_active Ceased
  • 2002-07-15 EP EP02748192A patent/EP1414485A1/en not_active Withdrawn
  • 2002-07-15 JP JP2003511857A patent/JP2004521952A/ja active Pending
  • 2002-07-15 US US10/196,437 patent/US20030095983A1/en not_active Abandoned
• 2004
  • 2004-01-06 ZA ZA200400060A patent/ZA200400060B/en unknown
• 2005
  • 2005-10-19 US US11/253,881 patent/US20060035818A1/en not_active Abandoned

Patent Citations (3)

Also Published As

Similar Documents

Legal Events