Optimized clinical performance of growth hormone with an expanded genetic code

Cho, Ho; Daniel, T. Sheidy; Buechler, Ying Ji; Litzinger, David C.; Maio, Zhenwei; Putnam, Anna-Maria Hays; Kraynov, Vadim S.; Sim, Bee-Cheng; Bussell, Stuart; Javahishvili, Tsotne; Kaphle, Sami; Viramontes, Guillermo; Ong, M M; Chu, Shaoyan; Gc, Becky; Lieu, Ricky; Knudsen, Nick; Castiglioni, Paola; Norman, Thea; Axelrod, Douglas W.; Hoffman, Andrew R.; Schultz, Peter G.; DiMarchi, Richard D.; Kimmel, Bruce E.

doi:10.1073/pnas.1100387108

Cited by 192 publications

(194 citation statements)

References 23 publications

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“…The estimated terminal t 1/2 after i.v. injection is 34 min for Genotropin, which is consistent with published results (24), and 47 h for hGH-CDR3H-coil-Herceptin (Fig. 2C), based on data fit using a two-compartment model.…”

Section: Resultssupporting

confidence: 79%

“…These data indicate that the hGH-CDR3H fusion folds correctly and is functional. In contrast, many of the long-acting forms of growth hormone in development, such as PEGylated GH and the XTEN and GH-GHR fusions, have a significant loss of in vitro potency (24)(25)(26).…”

Section: Resultsmentioning

confidence: 99%

“…1E). Finally, hGH-induced signal transduction was examined with a STAT5 phosphorylation assay in IM9 cells, a human B lymphoblastoid cell line naturally expressing hGHR (24). Upon treatment with hGH or hGH-BLV1H12, the hGHR signaling pathway is activated and results in STAT5 phosphorylation, which was quantified by flow cytometry analysis to give EC 50 values of 0.43 ± 0.02 and 0.49 ± 0.09 nM, respectively (Fig.…”

Section: Resultsmentioning

confidence: 99%

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Functional human antibody CDR fusions as long-acting therapeutic endocrine agonists

Liu

Wang

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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On the basis of the 3D structure of a bovine antibody with a wellfolded, ultralong complementarity-determining region (CDR), we have developed a versatile approach for generating human or humanized antibody agonists with excellent pharmacological properties. Using human growth hormone (hGH) and human leptin (hLeptin) as model proteins, we have demonstrated that functional human antibody CDR fusions can be efficiently engineered by grafting the native hormones into different CDRs of the humanized antibody Herceptin. The resulting Herceptin CDR fusion proteins were expressed in good yields in mammalian cells and retain comparable in vitro biological activity to the native hormones. Pharmacological studies in rodents indicated a 20-to 100-fold increase in plasma circulating half-life for these antibody agonists and significantly extended in vivo activities in the GH-deficient rat model and leptin-deficient obese mouse model for the hGH and hLeptin antibody fusions, respectively. These results illustrate the utility of antibody CDR fusions as a general and versatile strategy for generating long-acting protein therapeutics.antibody | protein engineering | growth hormone | leptin | pharmacology M any endocrine hormones are used therapeutically (1); however, they generally suffer from short circulating halflives (2, 3) and therefore require high doses and frequent injections to achieve efficacious exposures. For example, human growth hormone (hGH) is a 22-kDa four-helix-bundle protein produced by the pituitary gland, and its recombinant form has long been used for the treatment of growth hormone deficiency (GHD). Due to the short in vivo half-life of hGH, which is on the timescale of minutes, current GHD therapy requires daily injection (4), resulting in lower patient compliance (5). Studies have shown that a continuous infusion of recombinant (r)hGH has a similar efficacy and safety profile as daily rhGH therapy (6), and thus there is considerable interest in the development of long-acting forms of hGH. The first and only approved long-acting hGH, Nutropin Depot, a sustained-release formulation based on a biodegradable polymer, was withdrawn due to manufacturing difficulties. Various forms of long-acting GH have since been generated: those currently in clinical development include LB03002 (microparticle suspension), NNC126-0083 (PEGylation), NNC0195-0092 (reversible albumin-binding GH derivative), MOD-4023 (CTP modification), ACP-001 (prodrug strategy), Albutropin (albumin fusion), and VRS-317 (XTEN fusion), and have been extensively reviewed elsewhere (5). However, challenges associated with heterogeneity, stability, immunogenicity, toxicity, and/or compromised potency could potentially limit their clinical utility (3, 7-10).Another short-lived hormone in which there is considerable clinical interest is human leptin, a 16-kDa four-helix-bundle protein that plays a central role in the homeostasis of body weight. Recombinant leptin has been approved for the treatment of leptindeficient lipodystrophy patients (11), and ...

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Section: Resultssupporting

confidence: 79%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

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Functional human antibody CDR fusions as long-acting therapeutic endocrine agonists

Liu

Wang

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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“…For example, hydroxylamines have been used to generate glycoprotein mimics 155 , to label G-protein-coupled receptors 156 , antibodies 157 and therapeutic proteins for increased pharmacokinetics 6 , and for dual protein tagging and formation of bifunctional antibodies 158,159 . However, despite its widespread use, the reactions of aldehydes and ketones suffer from a number of key drawbacks.…”

Section: Review Nature Communications | Doi: 101038/ncomms5740mentioning

confidence: 99%

“…Ideally, this should proceed with near total conversion to generate homogenous constructs [2][3][4] . The applications of modified proteins are many; they are as varied as the in vivo tracking of protein-fluorophore conjugates 5 to the polyethylene glycol (PEG)ylation of therapeutic proteins to reduce immunogenicity 6 , from the production of materials with novel properties 7 to probing the mechanism of pathological enzymes 8 .…”

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confidence: 99%

Selective chemical protein modification

2014

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Chemical and Genetic Modification

Farys¹,

Ginn²,

Badescu³

et al. 2013

Pharmaceutical Sciences Encyclopedia

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There are many natural post‐translational modifications of proteins (e.g., glycosylation, ubiquitination, acylation,and amino acid derivatization).Glycosylation is common for therapeutic proteins. Classes of proteins modified by glycosylation include monoclonal antibodies, blood factors, hematopoietic proteins, and cytokines. Excluding monoclonal antibodies, many proteins were, at least when they were first introduced to the clinic, designed to be used as a replacement protein for the corresponding endogenous protein. Maintaining control of the protein structure to be as close as possible to the structure of the endogeneous protein is therefore a key goal. In this review, we focus on (i) optimization of protein pharmacokinetics, (ii) improvement of protein properties to be used as medicines (this primarily includes reduction of immunogenic sequences and improvement of protein stability), (iii) addition of a therapeutic effect, and (iv) use of proteins as diagnostics.

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Optimized clinical performance of growth hormone with an expanded genetic code

Cited by 192 publications

References 23 publications

Functional human antibody CDR fusions as long-acting therapeutic endocrine agonists

Functional human antibody CDR fusions as long-acting therapeutic endocrine agonists

Selective chemical protein modification

Chemical and Genetic Modification

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