Quantitative methods for developing Fc mutants with extended half‐lives

Kamei, Daniel T.; Lao, Bert J.; Ricci, Margaret Speed; Deshpande, Rohini; Xu, Han; Tidor, Bruce; Lauffenburger, Douglas A.

doi:10.1002/bit.20624

Cited by 25 publications

(9 citation statements)

References 52 publications

(61 reference statements)

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“…This may simply lead to an increased mass action effect of blocking locally produced VEGF or may provide a mechanism for enhanced degradation of VEGF in the tumor. Previously, another Fc variant with improved human FcRn binding was shown to have slower degradation rate and improved recycling behavior than WT in vitro, in human tumor cells expressing FcRn (35). In a perhaps related finding, Dall'Acqua and colleagues (16) reported that the concentration of the anti-RSV variant M252Y/S254T/T256E in the bronchoalveolar lavage of cynomolgus monkeys increased proportionally with the rise of serum IgG level.…”

Section: Discussionmentioning

confidence: 95%

A Therapeutic Anti–VEGF Antibody with Increased Potency Independent of Pharmacokinetic Half-life

Yeung¹,

Wu²,

Reyes³

et al. 2010

Cancer Research

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Bevacizumab [Avastin; anti-vascular endothelial growth factor (VEGF) antibody] is an antiangiogenic IgG approved for treating patients with certain types of colon, breast, and lung cancer. In these indications, bevacizumab is administered every 2 to 3 weeks, prompting us to study ways to reduce the frequency of administration. Increasing affinity to neonatal Fc receptor (FcRn) may extend the pharmacokinetic half-life of an antibody, but the quantitative effect of FcRn affinity on clearance has not been clearly elucidated. To gain further insight into this relationship, we engineered a series of anti-VEGF antibody variants with minimal amino acid substitutions and showed a range of half-life improvements in primates. These results suggest that, if proven clinically safe and effective, a modified version of bevacizumab could potentially provide clinical benefit to patients on long-term anti-VEGF therapy through less-frequent dosing and improved compliance with drug therapy. Moreover, despite having half-life similar to that of wild-type in mice due to the speciesspecific FcRn binding effects, the variant T307Q/N434A exhibited superior in vivo potency in slowing the growth of certain human tumor lines in mouse xenograft models. These results further suggest that FcRn variants may achieve increased potency through unidentified mechanisms in addition to increased systemic exposure. Cancer Res; 70(8); 3269-77. ©2010 AACR.

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Section: Discussionmentioning

confidence: 95%

A Therapeutic Anti–VEGF Antibody with Increased Potency Independent of Pharmacokinetic Half-life

Yeung¹,

Wu²,

Reyes³

et al. 2010

Cancer Research

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“…It was beneficial to generate a Fc fusion protein because it has been shown that the presence of the Fc significantly increases the half-life of a protein, thereby improving its pharmacokinetics properties and dynamics in vivo (37). In addition, presentation of costimulation by Fc receptor -bearing cells may be a particularly effective method for immunotherapy.…”

Section: Discussionmentioning

confidence: 99%

Construction and Preclinical Characterization of Fc-mGITRL for the Immunotherapy of Cancer

Hu¹,

Arias²,

Sadun³

et al. 2008

Clinical Cancer Research

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Purpose:To provide proper costimulation required for effective cancerT-cell immunity, Fc-GITRL fusion proteins were generated for use in immunotherapy protocols. Experimental Design: Soluble fusion proteins consisting of the Fc fragment of immunoglobulin and the murine glucocorticoid-induced tumor necrosis factor^related receptor ligand (mGITRL) connected with different linkers were genetically engineered and tested for their potency in two BALB/c solid tumor models. Results: In vivo, construct #178-14 (-5aa, -linker) showed the best activity (>90% tumor reduction) at doses ranging from 5 to 25 Ag and was found to be intact by gel electrophoresis. Similar doses used with construct #175-2 (-linker) produced good but not as high tumor regression. Construct #5-1 (+linker), which was found to be relatively unstable by SDS gel electrophoresis, produced <60% tumor regression and required a higher dose (100 Ag) to produce optimal results.Survival curves showed that Fc-mGITRL treatment extended the life of 80% of tumor-bearing mice to >3 months compared with controls that died by day 40. T-cell depletion studies showed that CD8 + T cells play a major role in Fc-mGITRL immunotherapy, and tumors removed from Fc-mGITRL^and DTA-1^treated mice showed a significant influx of granzyme B + lymphocytes compared with controls. Finally,Tregulatory (Treg) cell assays showed that, unlike other Fc fusion proteins, all three Fc-mGITRL constructs profoundly suppressed Treg activity. Conclusions: These studies suggest that a stable, intact Fc-mGITRL fusion protein can provide missing costimulation for the immunotherapy of solid tumors. In addition, Fc-mGITRL may alter Treg activity to enhance its effectiveness for tumor immunotherapy.

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“…As a consequence, they may have extended serum half-lifes by endocytic salvage via neonatal FcR (FcRn), which may compensate for slow tissue penetration. [30][31][32][33][34][35] Furthermore, they may elicit antibody-dependent cell-mediated cytotoxicity or phagocytosis (ADCC or ADCP, respectively) via Fcg receptors (FcgRs), which is important for cancer biotherapeutics. 3,[36][37][38][39] Fully functional Fc domains may also confer the potential to trigger the classical pathway of complementdependent humoral response, and thereby elicit complementdependent cytotoxicity (CDC).…”

Section: Introductionmentioning

confidence: 99%

CODV-Ig, a universal bispecific tetravalent and multifunctional immunoglobulin format for medical applications

Steinmetz

Vallée

Beil

et al. 2016

mAbs

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Bispecific immunoglobulins (Igs) typically contain at least two distinct variable domains (Fv) that bind to two different target proteins. They are conceived to facilitate clinical development of biotherapeutic agents for diseases where improved clinical outcome is obtained or expected by combination therapy compared to treatment by single agents. Almost all existing formats are linear in their concept and differ widely in drug-like and manufacture-related properties. To overcome their major limitations, we designed cross-over dual variable Ig-like proteins (CODV-Ig). Their design is akin to the design of circularly closed repeat architectures. Indeed, initial results showed that the traditional approach of utilizing (G4S) x linkers for biotherapeutics design does not identify functional CODV-Igs. Therefore, we applied an unprecedented molecular modeling strategy for linker design that consistently results in CODV-Igs with excellent biochemical and biophysical properties. CODV architecture results in a circular self-contained structure functioning as a self-supporting truss that maintains the parental antibody affinities for both antigens without positional effects. The format is universally suitable for therapeutic applications targeting both circulating and membrane-localized proteins. Due to the full functionality of the Fc domains, serum half-life extension as well as antibody-or complement-dependent cytotoxicity may support biological efficiency of CODV-Igs. We show that judicious choice in combination of epitopes and paratope orientations of bispecific biotherapeutics is anticipated to be critical for clinical outcome. Uniting the major advantages of alternative bispecific biotherapeutics, CODV-Igs are applicable in a wide range of disease areas for fast-track multi-parametric drug optimization.

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Quantitative methods for developing Fc mutants with extended half‐lives

Cited by 25 publications

References 52 publications

A Therapeutic Anti–VEGF Antibody with Increased Potency Independent of Pharmacokinetic Half-life

A Therapeutic Anti–VEGF Antibody with Increased Potency Independent of Pharmacokinetic Half-life

Construction and Preclinical Characterization of Fc-mGITRL for the Immunotherapy of Cancer

CODV-Ig, a universal bispecific tetravalent and multifunctional immunoglobulin format for medical applications

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