Exceptionally Potent and Broadly Cross-Reactive, Bispecific Multivalent HIV-1 Inhibitors Based on Single Human CD4 and Antibody Domains

Chen, Weizao; Yang, Feng; Prabakaran, Ponraj; Ying, Tianlei; Wang, Yanping; Sun, Jianyuan; Macedo, Camila; Zhu, Zhongyu; He, Yuxian; Polonis, Victoria R.; Dimitrov, Dimiter S.

doi:10.1128/jvi.02566-13

Cited by 56 publications

(79 citation statements)

References 70 publications

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“…Other MD mutants also showed ELISA binding activity (EC 50 s, 1.5-2 nM) much higher than that (EC 50 , >1,000 nM) of MD although slightly lower than that (EC 50 , 1 nM) of MD13 likely due to relatively inefficient CH1-CK heterodimerization in the other MD mutants compared to MD13 (data not shown). Given that mD1.22 has nanomolar affinities for the Env, while m36.4 barely binds without sCD4, as demonstrated in our previous studies, 4,10,16 the results suggest that the purified MD13 protein should contain mD1.22-CH1. As expected, mass spectrometry confirmed the presence of mD1.22-CH1 with abundance comparable to that of m36.4-CK (Fig.…”

mentioning

confidence: 86%

“…4Dm2m neutralized all HIV-1 isolates tested in vitro with potency and breadth greater than those of representative bnAbs such as VRC01 and CD4-Ig, a clinically tested Fc-fusion protein of the two-domain sCD4 D1D2. 4 However, our preliminary study in mice showed that 4Dm2m had a markedly shorter in vivo half-life than CD4-Ig. In addition to the potential difference in stability and specificity of binding moieties between 4Dm2m and CD4-Ig, we hypothesized that the use of CH1-CK heterodimerization scaffold and multiple long (G4S)3 linkers in 4Dm2m might contribute to the reduced pharmacokinetics because CH1-CK heterodimerization had previously been shown to be inefficient 11 and unstable, 12,13 and the (G4S)3 linker may be sensitive to proteolysis.…”

Section: Introductionmentioning

confidence: 99%

“…7,8 4Dm2m is a bispecific multivalent fusion protein composed of the engineered single-domain sCD4 mD1.22 and antibody domain m36.4 that targets the coreceptor binding site of the Env gp120. 4,9,10 The epitope of m36.4 is fully formed or exposed only in the presence of CD4 and therefore, mD1.22 and m36.4 synergize with each other in binding and neutralizing HIV-1. Both binding moieties are linked to the human IgG1 constant region via the (G4S)3 linkers and use the heavy chain constant domain 1 (CH1) and kappa light chain constant domain (CK) for heterodimerization (Fig.…”

Section: Introductionmentioning

confidence: 99%

“…1,2 However, the virus' ability to constantly mutate enables it to generate variants capable of evading immune recognition necessitating treatment either in combination with other antibodies recognizing different epitopes or parallel treatment with antiretroviral drugs to control the infection. 3 Bispecific fusion proteins of soluble forms of the HIV-1 primary receptor CD4 (sCD4) with bnAbs 4,5 or mimetics of the HIV-1 coreceptor CCR5 6 have emerged as novel classes of promising HIV-1 inhibitors. The two moieties bind the viral envelope glycoproteins (Env) cooperatively and with high avidity, leading to exceptionally potent and broad neutralizing activity of the fusion proteins.…”

Section: Introductionmentioning

confidence: 99%

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Improving the CH1-CK heterodimerization and pharmacokinetics of 4Dm2m, a novel potent CD4-antibody fusion protein against HIV-1

Chen

Bardhi

Yang

et al. 2016

mAbs

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(2016) Improving the CH1-CK heterodimerization and pharmacokinetics of 4Dm2m, a novel potent CD4-antibody fusion protein against HIV-1, mAbs, 8:4, 761-774, DOI: 10.1080/19420862.2016 To link to this article: https://doi.org/10. 1080/19420862.2016 ABSTRACTWe previously described 4Dm2m, an exceptionally potent broadly neutralizing CD4-antibody fusion protein against HIV-1. It was generated by fusing the engineered single human CD4 domain mD1.22 to both the N and C termini of the human IgG1 heavy chain constant region and the engineered single human antibody domain m36.4, which targets the CD4-induced coreceptor binding site of the viral envelope glycoprotein, to the N terminus of the human antibody kappa light chain constant region via the (G4S)3 polypeptide linkers. However, therapeutic use of 4Dm2m was limited by its short in vivo half-life. Here, we show that a combination of three approaches have successfully increased the persistence of 4Dm2m in mice. First, to stabilize the scaffold, we enhanced heterodimerization between the heavy chain constant domain 1 (CH1) and kappa light chain constant domain (CK) by using structure-guided design and phage-display library technologies. Second, to address the possibility that long polypeptide linkers might render fusion proteins more susceptible to proteolysis, we shortened the (G4S)3 linkers or replaced them with the human IgG1 hinge sequence, which is naturally designed for both flexibility and stability. Third, we introduced two amino acid mutations into the crystallizable fragment (Fc) of the scaffold previously shown to increase antibody binding to the neonatal Fc receptor (FcRn) and prolong half-lives in vivo. Collectively, these approaches markedly increased the serum concentrations of 4Dm2m in mice while not affecting other properties of the fusion protein. The new 4Dm2m variants are promising candidates for clinical development to prevent or treat HIV-1 infection. To our knowledge, this is the first report on stabilized CH1-CK, which is potentially useful as a new heterodimerization scaffold for generation of bispecific and multispecific antibodies or proteins with a more favorable pharmacokinetic profile.

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confidence: 86%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Improving the CH1-CK heterodimerization and pharmacokinetics of 4Dm2m, a novel potent CD4-antibody fusion protein against HIV-1

Chen

Bardhi

Yang

et al. 2016

mAbs

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“…First, in addition to being able to encode combinations of bNAbs, which has been shown to prevent or delay viral escape, the larger carrying capacity of ADV vectors should support the delivery of the newly engineered bispecific and multivalent bNAbs, which have been reported to be 100-200 times more potent than conventional gp160-targeting bNAbs. 42,43 By encoding even more highly potent bNAbs, viremia may be fully suppressed for a longer period of time, reducing the chances of virological resistance or escape. Second, clinical safety is a major concern with any viral vector, and immunogenic effects have been observed, albeit rarely, for both ADV 44 and AAV.…”

Section: Discussionmentioning

confidence: 99%

Adenovirus-Vectored Broadly Neutralizing Antibodies Directed Against gp120 Prevent Human Immunodeficiency Virus Type 1 Acquisition in Humanized Mice

et al. 2015

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{The first three authors contributed equally to this manuscript.Despite nearly three decades of research, a safe and effective vaccine against human immunodeficiency virus type 1 (HIV-1) has yet to be achieved. More recently, the discovery of highly potent anti-gp160 broadly neutralizing antibodies (bNAbs) has garnered renewed interest in using antibody-based prophylactic and therapeutic approaches. Here, we encoded bNAbs in first-generation adenoviral (ADV) vectors, which have the distinctive features of a large coding capacity and ease of propagation. A single intramuscular injection of ADV-vectorized bNAbs in humanized mice generated high serum levels of bNAbs that provided protection against multiple repeated challenges with a high dose of HIV-1, prevented depletion of peripheral CD4 + T cells, and reduced plasma viral loads to below detection limits. Our results suggest that ADV vectors may be a viable option for the prophylactic and perhaps therapeutic use of bNAbs in humans. INTRODUCTIONSince its emergence more than three decades ago, human immunodeficiency virus type 1 (HIV-1) remains a pandemic, with more than 60 million infected individuals to date and more than 32 million acquired immunodeficiency syndrome (AIDS)-related deaths.1,2 Despite intense research efforts, a safe and effective vaccine remains elusive. At present, highly active antiretroviral therapy (HAART) constitutes the mainstay of treatment and has resulted in HIV-infected individuals with plasma viral RNA loads (VLs) below the limits of detection, increased peripheral CD4 + T cell counts, and decreased patient morbidity and mortality. Despite the improved quality of life, HAART has a number of limitations including high cost, drug toxicity and interactions, emergence of virus resistance, and the need for indefinite treatment, necessitating alternative therapeutic approaches.

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Virus Entry Inhibitors: Past, Present, and Future

Jiang

2022

Advances in Experimental Medicine and Biology

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Exceptionally Potent and Broadly Cross-Reactive, Bispecific Multivalent HIV-1 Inhibitors Based on Single Human CD4 and Antibody Domains

Cited by 56 publications

References 70 publications

Improving the CH1-CK heterodimerization and pharmacokinetics of 4Dm2m, a novel potent CD4-antibody fusion protein against HIV-1

Improving the CH1-CK heterodimerization and pharmacokinetics of 4Dm2m, a novel potent CD4-antibody fusion protein against HIV-1

Adenovirus-Vectored Broadly Neutralizing Antibodies Directed Against gp120 Prevent Human Immunodeficiency Virus Type 1 Acquisition in Humanized Mice

Virus Entry Inhibitors: Past, Present, and Future

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