Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 2 de 2
Filter
Add more filters










Database
Language
Publication year range
1.
Proc Natl Acad Sci U S A ; 111(5): 1766-71, 2014 Feb 04.
Article in English | MEDLINE | ID: mdl-24443552

ABSTRACT

Using an expanded genetic code, antibodies with site-specifically incorporated nonnative amino acids were produced in stable cell lines derived from a CHO cell line with titers over 1 g/L. Using anti-5T4 and anti-Her2 antibodies as model systems, site-specific antibody drug conjugates (NDCs) were produced, via oxime bond formation between ketones on the side chain of the incorporated nonnative amino acid and hydroxylamine functionalized monomethyl auristatin D with either protease-cleavable or noncleavable linkers. When noncleavable linkers were used, these conjugates were highly stable and displayed improved in vitro efficacy as well as in vivo efficacy and pharmacokinetic stability in rodent models relative to conventional antibody drug conjugates conjugated through either engineered surface-exposed or reduced interchain disulfide bond cysteine residues. The advantages of the oxime-bonded, site-specific NDCs were even more apparent when low-antigen-expressing (2+) target cell lines were used in the comparative studies. NDCs generated with protease-cleavable linkers demonstrated that the site of conjugation had a significant impact on the stability of these rationally designed prodrug linkers. In a single-dose rat toxicology study, a site-specific anti-Her2 NDC was well tolerated at dose levels up to 90 mg/kg. These experiments support the notion that chemically defined antibody conjugates can be synthesized in commercially relevant yields and can lead to antibody drug conjugates with improved properties relative to the heterogeneous conjugates formed by nonspecific chemical modification.


Subject(s)
Antibodies/metabolism , Immunoconjugates/metabolism , Pharmaceutical Preparations/chemical synthesis , Protein Engineering/methods , Animals , Antibodies/blood , Antibodies/chemistry , Antibodies/toxicity , Batch Cell Culture Techniques , CHO Cells , Cell Death/drug effects , Cell Line , Cricetinae , Cricetulus , Cysteine/metabolism , Humans , Immunoconjugates/chemistry , Immunoconjugates/pharmacokinetics , Immunoconjugates/toxicity , Pharmaceutical Preparations/blood , Pharmaceutical Preparations/chemistry , Protein Stability/drug effects , Rats
2.
J Pept Sci ; 18(6): 383-93, 2012 Jun.
Article in English | MEDLINE | ID: mdl-22565812

ABSTRACT

The identification of leptin as a mediator of body weight regulation provided much initial excitement for the treatment of obesity. Unfortunately, leptin monotherapy is insufficient in reversing obesity in rodents or humans. Recent findings suggest that amylin is able to restore leptin sensitivity and when used in combination with leptin enhances body weight loss in obese rodents and humans. However, as the uniqueness of this combination therapy remains unclear, we assessed whether co-administration of leptin with other weight loss-inducing hormones equally restores leptin responsiveness in diet-induced obese (DIO) mice. Accordingly, we report here the design and characterization of a series of site-specifically enhanced leptin analogs of high potency and sustained action that, when administered in combination with exendin-4 or fibroblast growth factor 21 (FGF21), restores leptin responsiveness in DIO mice after an initial body weight loss of 30%. Using either combination, body weight loss was enhanced compared with either exendin-4 or FGF21 monotherapy, and leptin alone was sufficient to maintain the reduced body weight. In contrast, leptin monotherapy proved ineffective when identical weight loss was induced by caloric restriction alone over a comparable time. Accordingly, we find that a hypothalamic counter-regulatory response to weight loss, assessed using changes in hypothalamic agouti related peptide (AgRP) levels, is triggered by caloric restriction, but blunted by treatment with exendin-4. We conclude that leptin re-sensitization requires pharmacotherapy but does not appear to be restricted to a unique signaling pathway. Our findings provide preclinical evidence that high activity, long-acting leptin analogs are additively efficacious when used in combination with other weight-lowering agents.


Subject(s)
Diet/adverse effects , Fibroblast Growth Factors/pharmacology , Leptin/analogs & derivatives , Leptin/pharmacology , Obesity/metabolism , Peptides/pharmacology , Venoms/pharmacology , Animals , Body Weight , Drug Combinations , Exenatide , Fibroblast Growth Factors/administration & dosage , Leptin/administration & dosage , Leptin/therapeutic use , Mice , Mice, Inbred C57BL , Mice, Obese , Models, Molecular , Obesity/chemically induced , Obesity/drug therapy , Peptides/administration & dosage , Polyethylene Glycols/chemistry , Venoms/administration & dosage
SELECTION OF CITATIONS
SEARCH DETAIL
...