WO2017189891A1 - Méthodes de traitement d'infections par le virus zika et de complications associées - Google Patents

Méthodes de traitement d'infections par le virus zika et de complications associées Download PDF

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WO2017189891A1
WO2017189891A1 PCT/US2017/029918 US2017029918W WO2017189891A1 WO 2017189891 A1 WO2017189891 A1 WO 2017189891A1 US 2017029918 W US2017029918 W US 2017029918W WO 2017189891 A1 WO2017189891 A1 WO 2017189891A1
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zika virus
subject
immune complex
polypeptide
amino acid
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PCT/US2017/029918
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English (en)
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Neil M. Bodie
Elliot Altman
Hyo Park
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Middle Tennessee State University
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
    • A61K38/10—Peptides having 12 to 20 amino acids
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/08—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses
    • C07K16/10—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses from RNA viruses
    • C07K16/1081—Togaviridae, e.g. flavivirus, rubella virus, hog cholera virus
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
    • C07K7/04—Linear peptides containing only normal peptide links
    • C07K7/08—Linear peptides containing only normal peptide links having 12 to 20 amino acids
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

  • Humoral immune responses are triggered when an antigen binds specifically to an antibody.
  • the combination of an antibody molecule and an antigen forms a small, relatively soluble immune complex.
  • Antigens either can be foreign substances, such as viral or bacterial polypeptides, or can be "self-antigens" such as polypeptides normally found in the human body.
  • the immune system normally distinguishes foreign antigens from self-antigens. "Autoimmune" disease can occur, however, when this system breaks down, such that the immune system turns upon the body and destroys tissues or organ systems as if they were foreign substances.
  • Larger immune complexes are more pathogenic than small, more soluble immune complexes.
  • the formation of large, relatively insoluble immune complexes can result from both the interaction of antibody molecules with antigen and the interaction of antibody molecules with each other. Such immune complexes also can result from interactions between antibodies in the absence of antigen.
  • Antibodies can prevent infections by coating viruses or bacteria, but otherwise are relatively harmless by themselves.
  • organ specific tissue damage can occur when antibodies combine with antigens and the resulting immune complexes bind to certain effector molecules in the body. Effector molecules are so named because they carry out the pathogenic effects of immune complexes. By inhibiting the formation of large, insoluble immune complexes, or by inhibiting the binding of immune complexes to effector molecules, the tissue damaging effects of immune complexes may be prevented.
  • polypeptides having amino acid sequences based on those set forth in SEQ ID NO:2 and SEQ ID NO:20 also referred to as ICI406 or
  • NB406 can bind specifically and with high affinity to the C H -C H 3 domain of immunoglobulin molecules, thus inhibiting the formation of insoluble immune complexes containing antibodies and antigens, and preventing the binding of such complexes to effector molecules.
  • This document provides such polypeptides, other C H 2-C H 3 binding compounds, compositions containing the polypeptides and/or compounds, and methods for using the polypeptides and compositions to inhibit immune complex formation and therapeutic use in treating viral infections.
  • the polypeptides may inhibit or prevent the binding of an Fc receptor to an immune complex that includes an antibody and an antigen. That is, polypeptides may inhibit or prevent the binding of a protein that could otherwise bind to the Fc-portion of an antibody, including an antibody bound to an antigen, to the Fc-portion of the antibody.
  • This document is based in part on the discovery that Zika virus and/or a protein of Zika virus, nonstructural protein 1 (NS1), can act as an Fc receptor.
  • NS1 nonstructural protein 1
  • this document features a method for inhibiting immune complex formation in a subject, the method comprising administering to the subject a composition comprising a purified polypeptide, the polypeptide comprising the amino acid sequence (Xaai) m -Cys-Ala-Xaa 2 -His- Leu-Gly-Glu-Leu-Val-Trp-Cys-Thr-(Xaa3) « (SEQ ID NO:60), wherein Xaai is any amino acid, Xaa 2 is Trp, Tyr or Phe, 5-Hydroxytrphophan (5-HTP), 5-hydroxytryptamine (5-HT), or another amino acid derivative, Xaa 3 is any amino acid, and m and n independently are 0, 1, 2, 3, 4, or 5, and wherein the immune complex formation is associated with a viral infection.
  • Xaai is any amino acid
  • Xaa 2 is Trp, Tyr or Phe
  • 5-Hydroxytrphophan (5-HTP) 5-hydroxy
  • the immune complex formation can be associated with infection by Zika virus, associated with Antibody Dependent Enhancement (ADE) of Zika virus disease, or associated with Zika virus-associated Guillain-Barre syndrome (GBS).
  • ADE Antibody Dependent Enhancement
  • GFS Zika virus-associated Guillain-Barre syndrome
  • the immune complex formation can contribute to the enhancement of a Zika virus infection or can contribute to the transmission of a Zika virus infection.
  • the peptide can cause clinical or histological improvement of a Zika virus infection and/or Zika virus-associated Guillain-Barre syndrome (GBS).
  • the peptide can cause an improvement in or delay the onset of one or more of the histological characteristics of a Zika virus infection.
  • the peptide can cause an improvement in or delay the onset of one or more of the histological characteristics or symptoms of a neurological disorder and, in some aspects, a Zika-related neurological disorder.
  • the neurological disorder can include Guillain-Barre syndrome (GBS) or microcephaly.
  • the peptide can decrease the ADE of a Zika virus infection.
  • the peptide can cause an inhibition in or delay in the vertical transmission of Zika virus from a pregnant subject to a fetus.
  • the subject may have been diagnosed as having or may be suspected of having a Zika virus infection.
  • the Zika virus infection could be caused by an African lineage Zika virus or an Asian lineage Zika virus.
  • the subject may be exhibiting symptoms of a Zika infection, the subject may be at risk of contracting a Zika infection, and/or the subject may have been exposed to Zika virus infection.
  • the subject may be a human. The subject may be pregnant, suspected of being pregnant, or trying to become pregnant.
  • the subject may have or be suspected of having a coinfection with a pathogen other the Zika virus including, for example, human immunodeficiency virus (HIV), human cytomegalovirus (HCMV), herpes simplex virus (HSV), hepatitis C virus, human papilloma virus (HPV), Mycobacterium tuberculosis,
  • a pathogen other the Zika virus including, for example, human immunodeficiency virus (HIV), human cytomegalovirus (HCMV), herpes simplex virus (HSV), hepatitis C virus, human papilloma virus (HPV), Mycobacterium tuberculosis,
  • the subject may have or be suspected of having an autoimmune disease.
  • the peptide can inhibit immune complex formation.
  • the immune complex can be capable of transcytosis, including, for example, FcRn-mediated transcytosis.
  • the transcytosis can include, for example, placental transcytosis, blood brain barrier transcytosis, retinal transcytosis, epithelial cell transcytosis, endothelial cell transcytosis, and/or testicular transcytosis.
  • the immune complex formation can include Fc-mediated immune complex formation.
  • the immune complex formation can include binding of to an Fc Receptor (FcR), FcRn, DC-SIGN, mClq, or sClq.
  • the immune complex formation can include binding to an IgG IC.
  • the immune complex formation can include binding of an immune complex (IC), including a heterologous IC and/or an IgG IC, to an FcR, mClq, or sClq.
  • IC immune complex
  • a heterologous immune complex can include a complex formed between a non-Zika virus antibody and a non-Zika virus antigen.
  • An immune complex can include a Zika virus antibody and a Zika virus antigen.
  • An immune complex can include a non-Zika virus antibody and a Zika virus antigen.
  • the immune complex formation can include binding of Zika virus and/or a Zika virus virion.
  • the immune complex formation can include binding of a Zika virus protein (e.g., Zika NS1).
  • the FcR can include DC-SIGN, FcRn, or a FcyR.
  • a FcyR can include, for example, Fcyl, Fcylla HI 31 allele, Fcylla R131 allele, FcyRIIb, FcyRIIc, FcyRIIIa, or Fcylllb.
  • a therapeutic antibody may be administered to the subject before, during, or after administration of a composition including the polypeptide.
  • the therapeutic antibody can include at least one antibody that activates the inhibitory FcyR, FcyRII, or can include an antibody to FcRn or DC-SIGN.
  • this document describes a method for treatment of a disease or condition in a subject comprising inhibiting the interaction of DC-SIGN and an immune complex.
  • Inhibiting the interaction of DC-SIGN and an immune complex can include administering to the subject a composition comprising a polypeptide.
  • the disease or condition can include a viral disease or infection including, for example, a Zika virus infection.
  • the immune complex can include, an Fc-mediated immune complex.
  • the immune complex can include a Zika virus, a Zika virus protein, or an antigen derived from a Zika virus.
  • this document describes a method for treating Zika virus infection.
  • the method includes identifying a subject with a Zika virus infection or at risk of developing a Zika virus infection, and administering to the subject a composition comprising a polypeptide.
  • the polypeptide can comprise a terminal stabilizing group.
  • the terminal stabilizing group can be at the amino terminus of the polypeptide and can be a tripeptide having the amino acid sequence Xaa-Pro-Pro, wherein Xaa is any amino acid (e.g., Ala).
  • the terminal stabilizing group can be at the carboxy terminus of the polypeptide and can be a tripeptide having the amino acid sequence Pro-Pro-Xaa, wherein Xaa is any amino acid (e.g., Ala).
  • the polypeptide can comprise the amino acid sequence Xaa-Pro-Pro-Asp-Cys-Ala-Trp-His-Leu- Gly-Glu-Leu-Val-Trp-Cys-Thr (SEQ ID NO: 19), wherein Xaa is any amino acid.
  • the polypeptide can comprise the amino acid sequence Ala-Pro-Pro- Asp-Cys-Ala-Trp-His-Leu-Gly- Glu-Leu-Val-Trp-Cys-Thr (SEQ ID NO:20).
  • the polypeptide can comprise the amino acid sequence
  • FIG. 1 shows an in silco model of a secreted Zika nonstructural protein 1 (NS1) hexamer bound to an IgG hexamer.
  • polypeptides and other compounds capable of interacting with the C H 2- C H 3 cleft of an immunoglobulin molecule, such that interaction of the immunoglobulin with other molecules (e.g., effectors or other immunoglobulins) is blocked.
  • Methods for identifying such polypeptides and other compounds also are described, along with compositions and articles of manufacture containing the polypeptides and compounds.
  • this document provides methods for using the polypeptides and compounds to inhibit immune complex formation and to treat diseases (e.g., viral diseases such as Zika Virus Disease) in which IgG immune complexes bind to effector molecules, such as membrane bound Clq (mClq), soluble Clq (sClq), and FcyRs (including, but not limited to, FcyRI (and isoforms of FcyRs), FcyRIIa, FcyRIIb/c, FcyRIIIa, FcyRIIIb, and FcRn).
  • diseases e.g., viral diseases such as Zika Virus Disease
  • IgG immune complexes bind to effector molecules, such as membrane bound Clq (mClq), soluble Clq (sClq), and FcyRs (including, but not limited to, FcyRI (and isoforms of FcyRs), FcyRIIa, FcyRIIb/c, FcyRIIIa, F
  • This document further provides methods of treating a subject at risk of having a Zika virus infection, diagnosed as having a Zika virus infection, or suspected of having a Zika virus infection.
  • Zika virus and/or a protein of Zika virus can act as a viral Fc Receptor.
  • the Fc receptor (FcyR) activity of these viruses can facilitate interactions between the virus and a host cell receptor.
  • an "Fc receptor” refers to a protein that can bind to the Fc-portion of an antibody, including an antibody bound to an antigen and/or an antibody that is not bound to an antigen.
  • the Fc receptor (FcR) activity of these viruses can facilitate interactions between the virus and a host cell receptor.
  • an “immune complex” or “IC” refers both a complex between an immunoglobulin and an antigen and a complex between an immunoglobulin Fc region and other antibodies or factors.
  • an “Fc-mediated immune complex” refers to a complex between an immunoglobulin Fc region and other antibodies or factors including, for example, an Fc receptor.
  • the Fc-mediated immune complex may, optionally, include bound antigen.
  • the terms “heterologous immune complex” and “heterologous IC” mean a complex formed between a non-Zika virus antibody and a non-Zika virus antigen.
  • An immunoglobulin that is bound to both a host cell Fc receptor and a viral Fc receptor can enhance binding and/or entry of a virus into to a host cells. Because the interaction between the host cell and the virus is facilitated by interactions with the Fc-portion of the antibody in an immunoglobulin complex, a heterologous immune complex, not just an anti -viral immune complex, can bridge a host cell-virus interaction.
  • the previously unreported and surprising ability of Zika virus to interact with heterologous immune complexes may contribute to accelerated disease progression and increased infectivity, particularly in subjects having an increased number of circulating immune complexes.
  • an immunoglobulin molecule including, for example, SEQ ID NO: 19, SEQ ID NO:20, or SEQ ID NO:2
  • the interaction of the immunoglobulin with other molecules including a host cell Fc receptor and/or a viral Fc receptor can be blocked or abrogated.
  • immunoglobulin is part of an antibody-antigen immune complex, such abrogation can interfere with the immune complex's ability to bridge virus-host cell interactions.
  • immunoglobulins make up a class of proteins found in plasma and other bodily fluids that exhibit antibody activity and bind to other molecules (e.g., antigens and certain cell surface receptors) with a high degree of specificity. Based on their structure and biological activity, immunoglobulins can be divided into five classes: IgM, IgG, IgA, IgD, and IgE. IgG is the most abundant antibody class in the body; this molecule assumes a twisted "Y" shape configuration. With the exception of the IgMs, immunoglobulins are composed mainly of four peptide chains that are linked by several intrachain and interchain disulfide bonds.
  • the IgGs are composed of two polypeptide heavy chains (H chains) and two polypeptide light chains (L chains), which are coupled by disulfide bonds and non-covalent bonds to form a protein molecule with a molecular weight of approximately 150,000 daltons (Saphire et al. (2001) "Crystal Structure of a Neutralizing Human IgG against HIV-1 : A Template for Vaccine Design," Science, 293 : 1155-1159).
  • the average IgG molecule contains approximately 4.5 interchain disulfide bonds and approximately 12 intrachain disulfide bonds (Frangione and Milstein (1968) J. Mol. Biol. 33 :893-906).
  • the light and heavy chains of immunoglobulin molecules are composed of constant regions and variable regions (see, e.g., Padlan (1994) Mol. Immunol. 31 : 169-217).
  • the light chains of an IgGl molecule each contain a variable domain (V L ) and a constant domain (C L ).
  • the heavy chains each have four domains: an amino terminal variable domain (V H ), followed by three constant domains (C H I , C H 2, and the carboxy terminal C H 3).
  • a hinge region corresponds to a flexible junction between the C H I and C H 2 domains.
  • Papain digestion of an intact IgG molecule results in proteolytic cleavage at the hinge and produces an Fc fragment that contains the C H 2 and C H 3 domains, and two identical Fab fragments that each contain a C H I , C L , V H , and V L domain.
  • the Fc fragment has complement- and tissue-binding activity, while the Fab fragments have antigen-binding activity.
  • Immunoglobulin molecules can interact with other polypeptides through various regions. The majority of antigen binding, for example, occurs through the V L /V H region of the Fab fragment.
  • the hinge region also is thought to be important, as immunological dogma states that the binding sites for Fc receptors (FcR) are found in the hinge region of IgG molecules (see, e.g., Raghavan and Bjorkman (1996) Annu. Rev. Dev. Biol. 12: 181-200). More recent evidence, however, suggests that FcR interacts with the hinge region primarily when the immunoglobulin is monomelic (i.e., not immune-complexed). Such interactions typically involve the amino acids at positions 234-237 of the Ig molecule (Wiens et al. (2000) J. Immunol. 164:5313-5318).
  • Immunoglobulin molecules also can interact with other polypeptides through a cleft within the C H 2-C H 3 domain.
  • the "C H 2-C H 3 cleft" typically includes the amino acids at positions 251-255 within the Cm domain and the amino acids at positions 424-436 within the C H 3 domain.
  • numbering is with respect to an intact IgG molecule as in Kabat et al. (Sequences of Proteins of Immunological Interest, 5 th ed., Public Health Service, U.S. Department of Health and Human Services, Bethesda, MD).
  • the corresponding amino acids in other immunoglobulin classes can be readily determined by those of ordinary skill in the art.
  • the C H -C H 3 cleft is unusual in that it is characterized by both a high degree of solvent accessibility and a predominantly hydrophobic character, suggesting that burial of an exposed hydrophobic surface is an important driving force behind binding at this site.
  • a three- dimensional change occurs at the IgG C H -C H 3 cleft upon antigen binding, allowing certain residues (e.g., a histidine at position 435) to become exposed and available for binding.
  • residues e.g., a histidine at position 435
  • Antigen binding therefore can be important for determining whether an immunoglobulin binds to other molecules through the hinge or the Fc C H 2-C H 3 region.
  • the Fc region can bind to a number of effector molecules and other proteins, including the following:
  • FcRn The neonatal Fc receptor determines the half-life of the antibody molecule in the general circulation (Leach et al. (1996) J. Immunol., 157:3317-3322; Gheti and Ward (2000) Ann. Rev. Immunol., 18:739-766). Mice genetically lacking FcRn are protected from the deleterious effects of pathogenic autoantibodies due to the shortened half-life of the autoantibodies (Liu et al. (1997) J. Exp. Med., 186:777-783).
  • the only binding site of FcRn to the IgG Fc is the IgG Fc C H 2-C H 3 cleft and HIS 435 has been shown by 3D structure and alanine scan to be essential to FcRn to IgG Fc binding (Shields et al. (2001) J. Biol. Chem., 276:6591-6604; Martin et al. (2001) Mol. Cell, 7:867-877). Since the peptides described herein bind with high affinity to the C H 2-C H 3 cleft and HIS 435, the peptides are direct inhibitors of (immune complexed) IgG Fc to FcRn binding. An inhibitor of FcRn binding to immune complexes or to pathogenic autoantibodies would be useful in treating diseases involving pathogenic autoantibodies and/or immune complexes.
  • FcR The cellular Fc Receptor provides a link between the humoral immune response and cell-mediated effector systems
  • the Fey. Receptors are specific for IgG molecules, and include FcyRI, FcyRIIa, FcyRIIb/c, and FcyRIIIa/b (and alleles, phenotypes and genotypes thereof). These isotypes bind with differing affinities to monomeric and immune-complexed IgG.
  • Clq The first component of the classical complement pathway is CI, which exists in blood serum as a complex of three proteins, Clq, Clr, and Cls.
  • the classical complement pathway is activated when Clq binds to the Fc regions of antigen-bound IgG or IgM. Although the binding of Clq to a single Fc region is weak, Clq can form tight bonds to a cluster of Fc regions (Diebolder et al. (2014) Science, 343(6176): 1260-1263).
  • Fc-mediated immune complex formation The formation of immune complexes via interactions between immunoglobulin Fc regions and other antibodies or other factors (e.g., those described above) is referred to herein as "Fc- mediated immune complex formation” or “the Fc-mediated formation of an immune complex.” Immune complexes containing such interactions are termed “Fc-mediated immune complexes.”
  • Fc-mediated immune complexes can include immunoglobulin molecules with or without bound antigen, and typically include C H 2-C H 3 cleft-specific ligands that have higher binding affinity for immune complexed antibodies than for monomeric antibodies.
  • polypeptide is any chain of amino acid residues, regardless of post- translational modification (e.g., phosphorylation or glycosylation).
  • the polypeptides provided herein typically are between 10 and 50 amino acids in length (e.g., 10, 11, 12, 13, 14, 15, 20, 25, 30, 35, 40, 45, or 50 amino acids in length).
  • Polypeptides that are between 10 and 20 amino acids in length e.g., 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acids in length
  • the amino acid sequences of the polypeptides provided herein are somewhat constrained, but can have some variability.
  • the polypeptides provided herein typically include the amino acid sequence
  • Xaai can be absent or can be any amino acid (e.g., Arg or Asp).
  • Xaa 2 can be Phe, Tyr, Tip, 5- Hydroxytryptophan (5-HTP), or Arg.
  • Xaa 3 can be any amino acid.
  • Xaa 4 can be Gly or Ala, while Xaa 5 can be Glu or Ala.
  • Xaa 6 also can be absent or can be any amino acid (SEQ ID NO:57).
  • a polypeptide can include the amino acid sequence Asp-Cys-Ala-Trp-His-
  • polypeptides provided herein can be modified for use in vivo by the addition, at the amino- or carboxy -terminal end, of a stabilizing agent to facilitate survival of the polypeptide in vivo. This can be useful in situations in which peptide termini tend to be degraded by proteases prior to cellular uptake.
  • a stabilizing agent to facilitate survival of the polypeptide in vivo.
  • Such blocking agents can include, without limitation, additional related or unrelated peptide sequences that can be attached to the amino- and/or carboxy-terminal residues of the polypeptide (e.g., an acetyl group attached to the N-terminal amino acid or an amide group attached to the C-terminal amino acid).
  • attachment can be achieved either chemically, during the synthesis of the polypeptide, or by recombinant DNA technology using methods familiar to those of ordinary skill in the art.
  • blocking agents such as pyroglutamic acid or other molecules known in the art can be attached to the amino- and/or carboxy-terminal residues, or the amino group at the amino terminus or the carboxy group at the carboxy terminus can be replaced with a different moiety.
  • polypeptides provided herein can have a Pro-Pro-Xaa (e.g., Pro-Pro- Ala) sequence at their carboxy termini.
  • a polypeptide can include the amino acid sequence Cys-Ala-Trp-
  • polypeptides provided herein can include additional amino acid sequences at the amino terminus of the sequence set forth in SEQ ID NO: 1, the carboxy terminus of the sequence set forth in SEQ ID NO: 1, or both.
  • a polypeptide can contain the amino acid sequence
  • Xaai can be absent or can be any amino acid (e.g., Arg or Asp); Xaa 2 can be Phe, Tyr, 5-HTP, Trp, or Arg; Xaa 3 can be any amino acid; Xaa 4 can be Gly or Ala; Xaa 5 can be Glu or Ala; and Xaa 6 can be absent or can be any amino acid (SEQ ID NO: 58).
  • a polypeptide can include the amino acid sequence Trp-Glu-Ala-Asp-Cys-Ala-Xaa-His-Leu-Gly-Glu-Leu-Val-Trp-Cys-Thr-Lys-Val-Glu- Glu (SEQ ID NO:50), where Xaa is Arg, Trp, 5-HTP, Tyr, or Phe.
  • the amino acid sequences of the polypeptides described herein typically contain two cysteine residues. Polypeptides containing these amino acid sequences can cyclize due to formation of a disulfide bond between the two cysteine residues.
  • a person having ordinary skill in the art can, for example, use Ellman's Reagent to determine whether a peptide containing multiple cysteine residues is cyclized.
  • these cysteine residues can be substituted with other natural or non-natural amino acid residues that can form lactam bonds rather than disulfide bonds. For example, one cysteine residue could be replaced with aspartic acid or glutamic acid, while the other could be replaced with ornithine or lysine.
  • a lactam bridge By varying the amino acids that form a lactam bridge, a polypeptide provided herein can be generated that contains a bridge approximately equal in length to the disulfide bond that would be formed if two cysteine residues were present in the polypeptide.
  • the polypeptides provided herein can contain an amino acid tag.
  • a "tag” is generally a short amino acid sequence that provides a ready means of detection or purification through
  • tags such as c-myc, hemagglutinin, polyhistidine, or FLAG (an eight amino acid peptide tag; Sigma-Aldrich Corp., St. Louis, MO) can be used to aid
  • a polypeptide with a polyhistidine tag can be purified based on the affinity of histidine residues for nickel ions (e.g., on a Ni-NTA column), and can be detected in western blots by an antibody against polyhistidine (e.g., the Penta-His antibody; Qiagen, Valencia, CA).
  • Tags can be inserted anywhere within the polypeptide sequence, although insertion at the amino- or carboxy -terminus is particularly useful.
  • amino acid refers to natural amino acids, unnatural amino acids, and amino acid analogs, all in their D and L stereoisomers if their structures so allow. Natural amino acids include alanine (Ala), arginine (Arg), asparagine (Asn), aspartic acid (Asp), cysteine (Cys), glutamine (Gin), glutamic acid (Glu), glycine (Gly), histidine (His), isoleucine (He), leucine (Leu), lysine (Lys), methionine (Met), phenylalanine (Phe), proline (Pro), serine (Ser), threonine (Thr), tryptophan (Tip), tyrosine (Tyr), and valine (Val).
  • Natural amino acids include alanine (Ala), arginine (Arg), asparagine (Asn), aspartic acid (Asp), cysteine (Cys), glutamine (Gin), glutamic acid (Glu
  • Unnatural amino acids include, but are not limited to 5-Hydroxytryptophan, azetidinecarboxylic acid, 2-aminoadipic acid, 3- aminoadipic acid, beta-alanine, aminopropionic acid, 2-aminobutyric acid, 4-aminobutyric acid, 6-aminocaproic acid, 2-aminoheptanoic acid, 2-aminoisobutyric acid, 3-aminoisobutyric acid, 2- aminopimelic acid, 2,4-diaminoisobutyric acid, desmosine, 2,2'-diaminopimelic acid, 2,3- diaminopropionic acid, N-ethylglycine, N-ethylasparagine, hydroxylysine, allo-hydroxylysine, 3- hydroxyproline, 4-hydroxyproline, isodesmosine, allo-isoleucine, N-methylglycine, N- methylisoleucine, N-methylvaline, norvaline,
  • an “analog” is a chemical compound that is structurally similar to another but differs slightly in composition (as in the replacement of one atom by an atom of a different element or in the presence of a particular functional group).
  • An “amino acid analog” therefore is structurally similar to a naturally occurring amino acid molecule as is typically found in native polypeptides, but differs in composition such that either the C-terminal carboxy group, the N-terminal amino group, or the side-chain functional group has been chemically modified to another functional group.
  • Amino acid analogs include natural and unnatural amino acids which are chemically blocked, reversibly or irreversibly, or modified on their N-terminal amino group or their side- chain groups, and include, for example, methionine sulfoxide, methionine sulfone, S- (carboxymethyl)-cysteine, S-(carboxymethyl)-cysteine sulfoxide and S-(carboxymethyl)- cysteine sulfone.
  • Amino acid analogs may be naturally occurring, or can be synthetically prepared.
  • Non-limiting examples of amino acid analogs include 5-Hydroxytryptophan (5-HTP), aspartic acid-(beta-methyl ester), an analog of aspartic acid; N-ethylglycine, an analog of glycine; and alanine carboxamide, an analog of alanine
  • 5-HTP 5-Hydroxytryptophan
  • aspartic acid-(beta-methyl ester) an analog of aspartic acid
  • N-ethylglycine an analog of glycine
  • alanine carboxamide an analog of alanine
  • polypeptide backbone The stereochemistry of a polypeptide can be described in terms of the topochemical arrangement of the side chains of the amino acid residues about the polypeptide backbone, which is defined by the peptide bonds between the amino acid residues and the a-carbon atoms of the bonded residues.
  • polypeptide backbones have distinct termini and thus direction.
  • the majority of naturally occurring amino acids are L-amino acids.
  • Naturally occurring polypeptides are largely comprised of L-amino acids.
  • D-amino acids are the enantiomers of L-amino acids and can form peptides that are herein referred to as "inverso" polypeptides (i.e., peptides corresponding to native peptides but made up of D-amino acids rather than L-amino acids).
  • a "retro" polypeptide is made up of L-amino acids, but has an amino acid sequence in which the amino acid residues are assembled in the opposite direction of the native peptide sequence.
  • Retro-inverso modification of naturally occurring polypeptides involves the synthetic assembly of amino acids with a-carbon stereochemistry opposite to that of the corresponding L-amino acids (i.e., D- or D-allo-amino acids), in reverse order with respect to the native polypeptide sequence.
  • a retro-inverso analog thus has reversed termini and reversed direction of peptide bonds, while approximately maintaining the topology of the side chains as in the native peptide sequence.
  • the term “native” refers to any sequence of L-amino acids used as a starting sequence for the preparation of partial or complete retro, inverso or retro-inverso analogs.
  • Partial retro-inverso polypeptide analogs are polypeptides in which only part of the sequence is reversed and replaced with enantiomeric amino acid residues. Since the retro-inverted portion of such an analog has reversed amino and carboxyl termini, the amino acid residues flanking the retro-inverted portion can be replaced by side-chain-analogous a-substituted geminal- diaminomethanes and malonates, respectively. Alternatively, a polypeptide can be a complete retro-inverso analog, in which the entire sequence is reversed and replaced with D-amino acids.
  • Peptidomimetic compounds that are designed on the basis of the amino acid sequences of polypeptides.
  • Peptidomimetic compounds are synthetic, non-peptide compounds having a three-dimensional conformation (i.e., a "peptide motif,") that is
  • Peptidomimetic compounds can be designed to mimic any of the polypeptides provided herein.
  • Peptidomimetic compounds that are protease resistant are particularly useful.
  • peptidomimetic compounds may have additional characteristics that enhance therapeutic utility, such as increased cell permeability and prolonged biological half-life.
  • Such compounds typically have a backbone that is partially or completely non-peptide, but with side groups that are identical or similar to the side groups of the amino acid residues that occur in the peptide upon which the peptidomimetic compound is based.
  • Several types of chemical bonds e.g., ester, thioester, thioamide, retroamide, reduced carbonyl, dimethylene and ketomethylene
  • ester, thioester, thioamide, retroamide, reduced carbonyl, dimethylene and ketomethylene are known in the art to be useful substitutes for peptide bonds in the construction of peptidomimetic compounds.
  • K d is expressed as a concentration, with a low K d value (e.g., less than 100 nanomolar (nM)) signifying high affinity.
  • Polypeptides that can interact with an immunoglobulin molecule typically have a binding affinity of at least 1 micromolar ( ⁇ ) (e.g., at least 500 nM, at least 100 nM, at least 50 nM, or at least 10 nM) for the C H 2-C H 3 cleft of the immunoglobulin.
  • Polypeptides provided herein can bind with substantially equivalent affinity to immunoglobulin molecules that are bound by antigen and to monomeric immunoglobulins.
  • the polypeptides described herein can have a higher binding affinity (e.g., at least 10-fold, at least 100-fold, or at least 1000-fold higher binding affinity) for immunoglobulin molecules that are bound by antigen than for monomeric immunoglobulins.
  • Conformational changes that occur within the Fc region of an immunoglobulin molecule upon antigen binding to the Fab region are likely involved in a difference in affinity.
  • the crystal structures of bound and unbound NC6.8 Fab (from a murine monoclonal antibody) showed that the tail of the Fab heavy chain was displaced by 19 angstroms in crystals of the antigen/antibody complex, as compared to its position in unbound Fab (Guddat et al. (1994) J. Mol.
  • Immune-complexed (antigen-bound) IgG has a more open configuration and thus is more conducive to ligand binding.
  • the binding affinity of RF for immune-complexed IgG is much greater than the binding affinity of RF for monomeric IgG (Corper et al. (1997) Nat. Struct. Biol. 4:374; Sohi et al. (1996) Immunol. 88:636). The same typically is true for the polypeptides provided herein.
  • the polypeptides described herein can bind to the C H -C H 3 cleft of immunoglobulin molecules, they can be useful for blocking the interaction of other factors (e.g., FcRn, FcR, Clq, histones, MBP, SOD1 and other immunoglobulins) to the Fc region of the immunoglobulin, and thus can inhibit Fc-mediated immune complex formation.
  • inhibit is meant that Fc-mediated immune complex formation is reduced in the presence of a polypeptide provided herein, as compared to the level of immune complex formation in the absence of the polypeptide. Such inhibiting can occur in vitro (e.g., in a test tube) or in vivo (e.g., in an individual). Any suitable method can be used to assess the level of immune complex formation. Many such methods are known in the art, and some of these are described herein.
  • the polypeptides described herein typically interact with the C H -C H 3 cleft of an immunoglobulin molecules.
  • immunoglobulin molecule in a monomeric fashion (i.e., interact with only one immunoglobulin molecule and thus do not link two or more immunoglobulin molecules together) with a 1 :2 IgG Fc to peptide stoichiometry. Interactions with other immunoglobulin molecules through the Fc region therefore are precluded by the presence of the polypeptide.
  • the inhibition of Fc-mediated immune complex formation can be assessed in vitro, for example, by incubating an IgG molecule with a labeled immunoglobulin molecule (e.g., a fluorescently or enzyme (ELISA) labeled Fc Receptor or Clq in the presence and absence of a polypeptide, and measuring the amount of labeled immunoglobulin that is incorporated into an immune complex.
  • a labeled immunoglobulin molecule e.g., a fluorescently or enzyme (ELISA) labeled Fc Receptor or Clq
  • ELISA fluorescently or enzyme
  • Polypeptides can be produced by a number of methods, many of which are well known in the art.
  • a polypeptide can be obtained by extraction from a natural source (e.g., from isolated cells, tissues or bodily fluids), by expression of a recombinant nucleic acid encoding the polypeptide (as, for example, described below), or by chemical synthesis (e.g., by solid-phase synthesis or other methods well known in the art, including synthesis with an ABI peptide synthesizer; Applied Biosystems, Foster City, CA).
  • Methods for synthesizing retro-inverso polypeptide analogs (Bonelli et al. (1984) Int. J. Peptide Protein Res.
  • nucleic acid refers to both RNA and DNA, including cDNA, genomic DNA, and synthetic (e.g., chemically synthesized) DNA.
  • the nucleic acid can be double- stranded or single-stranded (i.e., a sense or an antisense single strand).
  • isolated as used herein with reference to a nucleic acid refers to a naturally-occurring nucleic acid that is not immediately contiguous with both of the sequences with which it is immediately contiguous (one at the 5' end and one at the 3' end) in the naturally-occurring genome of the organism from which it is derived.
  • isolated nucleic acids also includes any non-naturally-occurring nucleic acid sequence, since such non-naturally-occurring sequences are not found in nature and do not have immediately contiguous sequences in a naturally-occurring genome.
  • An isolated nucleic acid can be, for example, a DNA molecule, provided one of the nucleic acid sequences that is normally immediately contiguous with the DNA molecule in a naturally- occurring genome is removed or absent.
  • an isolated nucleic acid includes, without limitation, a DNA molecule that exists as a separate molecule (e.g., a chemically synthesized nucleic acid, or a cDNA or genomic DNA fragment produced by PCR or restriction
  • an isolated nucleic acid can include an engineered nucleic acid such as a recombinant DNA molecule that is part of a hybrid or fusion nucleic acid.
  • vectors containing the nucleic acids described herein are vectors containing the nucleic acids described herein.
  • a "vector” is a replicon, such as a plasmid, phage, or cosmid, into which another DNA segment may be inserted so as to bring about the replication of the inserted segment.
  • Polypeptides can be developed using phage display, for example. Methods well known to those skilled in the art may use phage display to develop the polypeptides described herein.
  • the vectors can be, for example, expression vectors in which the nucleotides encode the polypeptides provided herein with an initiator methionine, operably linked to expression control sequences.
  • operably linked means incorporated into a genetic construct so that expression control sequences effectively control expression of a coding sequence of interest.
  • An "expression control sequence” is a DNA sequence that controls and regulates the transcription and translation of another DNA sequence
  • an "expression vector” is a vector that includes expression control sequences, so that a relevant DNA segment incorporated into the vector is transcribed and translated.
  • a coding sequence is "operably linked" and “under the control” of transcriptional and translational control sequences in a cell when RNA polymerase transcribes the coding sequence into mRNA, which then is translated into the protein encoded by the coding sequence.
  • Expression vectors can be used in a variety of systems (e.g., bacteria, yeast, insect cells, and mammalian cells), as described herein.
  • suitable expression vectors include, without limitation, plasmids and viral vectors derived from, for example, herpes viruses, retroviruses, vaccinia viruses, adenoviruses, and adeno-associated viruses.
  • a wide variety of suitable expression vectors and systems are commercially available, including the pET series of bacterial expression vectors (Novagen, Madison, WI), the Adeno-X expression system (Clontech, Mountain View, CA), the Baculogold baculovirus expression system (BD Biosciences Pharmingen, San Diego, CA), and the pCMV-Tag vectors (Stratagene, La Jolla, CA).
  • Expression vectors that encode the polypeptides described herein can be used to produce the polypeptides.
  • Expression systems that can be used for small or large scale production of polypeptides include, without limitation, microorganisms such as bacteria (e.g., E. coli and B. subtilis) transformed with recombinant bacteriophage DNA, plasmid DNA, or cosmid DNA expression vectors containing the nucleic acid molecules provided herein; yeast (e.g., S.
  • yeast expression vectors containing the nucleic acid molecules of the invention
  • insect cell systems infected with recombinant virus expression vectors (e.g., baculovirus) containing the nucleic acid molecules provided herein
  • plant cell systems infected with recombinant virus expression vectors e.g., tobacco mosaic virus
  • recombinant plasmid expression vectors e.g., Ti plasmid
  • mammalian cell systems e.g., primary cells or immortalized cell lines such as COS cells, CHO cells, HeLa cells, HEK 293 cells, and 3T3 LI cells harboring recombinant expression constructs containing promoters derived from the genome of mammalian cells (e.g., the metallothionein promoter) or from mammalian viruses (e.g., the adenovirus late promoter and the cytomegalovirus promote
  • purified polypeptide refers to a polypeptide that either has no naturally occurring counterpart (e.g., a peptidomimetic), or has been chemically synthesized and is thus uncontaminated by other polypeptides, or that has been separated or purified from other cellular components by which it is naturally accompanied (e.g., other cellular proteins, polynucleotides, or cellular components).
  • the polypeptide is considered “purified” when it is at least 70%, by dry weight, free from the proteins and naturally occurring organic molecules with which it naturally associates.
  • a preparation of purified polypeptide therefore can be, for example, at least 80%, at least 90%, or at least 99%, by dry weight, the polypeptide.
  • Suitable methods for purifying polypeptides can include, for example, affinity chromatography, immunoprecipitation, size exclusion chromatography, and ion exchange chromatography. The extent of purification can be measured by any appropriate method, including but not limited to: column
  • This document provides methods for designing, modeling, and identifying compounds that can bind to the C H -C H 3 cleft of an immunoglobulin molecule and thus serve as inhibitors of Fc- mediated immune complex formation.
  • Such compounds also are referred to herein as "ligands.”
  • Compounds designed, modeled, and identified by these methods typically can interact with an immunoglobulin molecule through the C H 2-C H 3 cleft, and typically have a binding affinity of at least 1 ⁇ (e.g., at least 500 nM, at least 100 nM, at least 50 nM, or at least 10 nM) for the C H 2- C H 3 cleft of the immunoglobulin.
  • Such compounds generally have higher binding affinity (e.g., at least 10-fold, at least 100-fold, or at least 1000-fold higher binding affinity) for immune- complexed immunoglobulin molecules than for monomeric immunoglobulin molecules.
  • Compounds typically interact with the C H 2-C H 3 cleft of an immunoglobulin molecule in a monomeric fashion (i.e., interact with only one immunoglobulin molecule and thus do not link two or more immunoglobulin molecules together).
  • the interactions between a compound and an immunoglobulin molecule typically involve the amino acid residues at positions 252, 253, 435, and 436 of the immunoglobulin (number according to Kabat, supra).
  • SEQ ID NO:20 may have hydrophobic packing with IgG Fc Met-252, Ile-253, Ser-254, His-435 and Tyr-436 (e.g., the indole ring of Trp-14 in SEQ ID NO:20 can have a hydrophobic interaction with IgG Fc His-435).
  • Alanine substitution of IgG Fc Asn-434, His-435 or Tyr-436 can disrupt binding (AAG>1.5 kcal/mol).
  • alanine substitution of SEQ ID NO:20 Val- 13 or Trp-14 can result in disruption of binding (AAG>2.0 kcal/mol).
  • the interaction between compounds and the C H 2-C H 3 cleft renders the compounds capable of inhibiting the Fc-mediated formation of immune complexes by blocking the binding of other factors (e.g., Fc:Fc interactions, FcyRs, FcRn, histones, MBP, MOG, RF, Tau protein, a- synuclein, SOD 1, TNF and Clq) to the C H 2-C H 3 cleft.
  • Compounds identified by the methods provided herein can be polypeptides such as, for example, those described herein.
  • a compound can be any suitable type of molecule that can specifically bind to the C H -C H 3 cleft of an immunoglobulin molecule.
  • modeling is meant quantitative and/or qualitative analysis of receptor-ligand
  • modeling typically is performed using a computer and may be further optimized using known methods.
  • Methods of designing ligands that bind specifically (i.e., with high affinity) to the C H -C H 3 cleft of an immunoglobulin molecule having bound antigen typically are computer-based, and involve the use of a computer having a program capable of generating an atomic model.
  • Computer programs that use X-ray crystallography data are particularly useful for designing ligands that can interact with an Fc C H 2-C H 3 cleft.
  • Programs such as RasMol for example, can be used to generate a three dimensional model of a C H 2-C H 3 cleft and/or determine the structures involved in ligand binding.
  • Methods can include, for example, providing to a computer the atomic structural coordinates for amino acid residues within the C H 2-C H 3 cleft (e.g., amino acid residues at positions 252, 253, 435, and 436 of the cleft) of an immunoglobulin molecule in an Fc-mediated immune complex, using the computer to generate an atomic model of the C H 2-C H 3 cleft, further providing the atomic structural coordinates of a candidate compound and generating an atomic model of the compound optimally positioned within the C H 2-C H 3 cleft, and identifying the candidate compound as a ligand of interest if the compound interacts with the amino acid residues at positions 252, 253, 435, and 436 of the cleft.
  • the data provided to the computer also can include the atomic coordinates of amino acid residues at positions in addition to 252, 253, 435, and 436.
  • “optimally positioned” is meant positioned to optimize hydrophobic interactions between the candidate compound and the amino acid residues at positions 252, 253, 435, and 436 of the C H 2- C H 3 cleft.
  • a method for designing a ligand having specific binding affinity for the C H 2-C H 3 cleft of an immunoglobulin molecule can utilize a computer with an atomic model of the cleft stored in its memory. The atomic coordinates of a candidate compound then can be provided to the computer, and an atomic model of the candidate compound optimally positioned can be generated.
  • a candidate compound can be identified as a ligand having specific binding affinity for the C H 2-C H 3 cleft of an immunoglobulin molecule if, for example, the compound interacts with the amino acid residues at positions 252, 253, 435, and 436 of the cleft.
  • Soluble Fcylla inhibits the binding of immune complexed (but not monomelic, non-immune complexed) IgG Fc to RF-AN (Wines et al. (2003) Immunol. 109:246-254), and inhibitors that bind to the IgG Fc C H 2-C H 3 cleft, such as the peptides described herein, inhibit the binding of immune complexed (antigen-bound) IgG Fc to FcyRs.
  • Compounds also can be interactively designed from structural information of the compounds described herein using other structure-based design/modeling techniques (see, e.g., Jackson (1997) Seminars in Oncology 24:L164-172; and Jones et al. (1996) J. Med. Chem. 39:904-917).
  • Compounds and polypeptides also can be identified by, for example, identifying candidate compounds by computer modeling as fitting spatially and preferentially (i.e., with high affinity) into the C H 2-C H 3 cleft of an immunoglobulin molecule, and then screening those compounds in vitro or in vivo for the ability to inhibit Fc-mediated immune complex formation. Suitable methods for such in vitro and in vivo screening include those described herein.
  • compositions and articles of manufacture that can be used in methods for treating conditions that arise from abnormal Fc-mediated immune complex formation (e.g., over-production of Fc-mediated immune complexes).
  • the polypeptides, compounds, and compositions provided herein can be administered to a subject (e.g., a human or another mammal) having an viral infection, for example, that can be alleviated by modulating Fc- mediated immune complex formation and inhibit immune complexed IgG Fc to mClq, sClq, FcyRs, FcRn, and/or DC-SIGN.
  • one or more polypeptides or compounds can be administered to a subject suspected of having a disease or condition associated with immune complex formation.
  • compositions generally contain one or more polypeptides and compounds described herein.
  • a C H -C H 3 binding polypeptide for example, can be in a pharmaceutically acceptable carrier or diluent, and can be administered in amounts and for periods of time that will vary depending upon the nature of the particular disease, its severity, and the subject's overall condition.
  • the polypeptide is administered in an inhibitory amount (i.e., in an amount that is effective for inhibiting the production of immune complexes in the cells or tissues contacted by the polypeptide).
  • the polypeptides and methods described herein also can be used prophylactically, e.g., to minimize immunoreactivity in a subject at risk for abnormal or overproduction of immune complexes (e.g., a transplant recipient).
  • the ability of a polypeptide to inhibit Fc-mediated immune complex formation can be assessed by, for example, measuring immune complex levels in a subject before and after treatment.
  • a number of methods can be used to measure immune complex levels in tissues or biological samples, including those that are well known in the art. If the subject is a research animal, for example, immune complex levels in the joints can be assessed by immunostaining following euthanasia.
  • the effectiveness of an inhibitory polypeptide also can be assessed by direct methods such as measuring the level of circulating immune complexes in serum samples. Alternatively, indirect methods can be used to evaluate the effectiveness of polypeptides in live subjects.
  • reduced immune complex formation can be inferred from clinical improvement of immune mediated diseases or in vitro or in vivo models of which have been shown to be essential in the therapeutic use in treating Atherosclerosis.
  • Methods for formulating and subsequently administering therapeutic compositions are well known to those skilled in the art. Dosing is generally dependent on the severity and
  • Optimum dosages can vary depending on the relative potency of individual polypeptides, and can generally be estimated based on EC50 found to be effective in in vitro and in vivo animal models. Typically, dosage is from 0.01 ug to 100 g per kg of body weight, and may be given once or more daily, biweekly, weekly, monthly, or even less often. Following successful treatment, it may be desirable to have the patient undergo maintenance therapy to prevent the recurrence of the disease state.
  • compositions and formulations that include the polypeptides and/or compounds described herein.
  • Polypeptides therefore can be admixed, encapsulated, conjugated or otherwise associated with other molecules, molecular structures, or mixtures of compounds such as, for example, liposomes, polyethylene glycol, receptor targeted molecules, or oral, rectal, topical or other formulations, for assisting in uptake, distribution and/or absorption.
  • a “pharmaceutically acceptable carrier” (also referred to herein as an “excipient”) is a pharmaceutically acceptable solvent, suspending agent, or any other pharmacologically inert vehicle for delivering one or more therapeutic compounds (e.g., C H 2-C H 3 binding polypeptides) to a subject.
  • Pharmaceutically acceptable carriers can be liquid or solid, and can be selected with the planned manner of administration in mind so as to provide for the desired bulk, consistency, and other pertinent transport and chemical properties, when combined with one or more of therapeutic compounds and any other components of a given pharmaceutical composition.
  • Typical pharmaceutically acceptable carriers that do not deleteriously react with amino acids include, by way of example and not limitation: water; saline solution; binding agents (e.g., polyvinylpyrrolidone or hydroxypropyl methylcellulose); fillers (e.g., lactose and other sugars, gelatin, or calcium sulfate); lubricants (e.g., starch, polyethylene glycol, or sodium acetate); disintegrates (e.g., starch or sodium starch glycolate); and wetting agents (e.g., sodium lauryl sulfate).
  • binding agents e.g., polyvinylpyrrolidone or hydroxypropyl methylcellulose
  • fillers e.g., lactose and other sugars, gelatin, or calcium sulfate
  • lubricants e.g., starch, polyethylene glycol, or sodium acetate
  • disintegrates e.g., starch or sodium starch glycolate
  • compositions can be administered by a number of methods, depending upon whether local or systemic treatment is desired and upon the area to be treated.
  • Administration can be, for example, topical (e.g., transdermal, sublingual, ophthalmic, or intranasal); pulmonary (e.g., by inhalation or insufflation of powders or aerosols); oral; or parenteral (e.g., by subcutaneous, intrathecal, intraventricular, intramuscular, or intraperitoneal injection, or by intravenous drip).
  • Administration can be rapid (e.g., by injection) or can occur over a period of time (e.g., by slow infusion or administration of slow release formulations).
  • C H 2-C H 3 binding polypeptides can be administered by injection or infusion into the cerebrospinal fluid, preferably with one or more agents capable of promoting penetration of the polypeptides across the blood-brain barrier.
  • Formulations for topical administration of C H 2-C H 3 binding polypeptides include, for example, sterile and non-sterile aqueous solutions, non-aqueous solutions in common solvents such as alcohols, or solutions in liquid or solid oil bases. Such solutions also can contain buffers, diluents and other suitable additives.
  • Pharmaceutical compositions and formulations for topical administration can include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids, and powders.
  • Nasal sprays are particularly useful, and can be administered by, for example, a nebulizer or another nasal spray device. Administration by an inhaler also is particularly useful. Conventional pharmaceutical carriers, aqueous, powder or oily bases, thickeners and the like may be necessary or desirable.
  • compositions and formulations for oral administration include, for example, powders or granules, suspensions or solutions in water or non-aqueous media, capsules, sachets, or tablets. Such compositions also can incorporate thickeners, flavoring agents, diluents, emulsifiers, dispersing aids, or binders. Compositions and formulations for parenteral, intrathecal or intraventricular administration can include sterile aqueous solutions, which also can contain buffers, diluents and other suitable additives (e.g., penetration enhancers, carrier compounds and other pharmaceutically acceptable carriers).
  • suitable additives e.g., penetration enhancers, carrier compounds and other pharmaceutically acceptable carriers.
  • compositions include, without limitation, solutions, emulsions, aqueous suspensions, and liposome-containing formulations. These compositions can be generated from a variety of components that include, for example, preformed liquids, self-emulsifying solids and self-emulsifying semisolids.
  • Emulsions are often biphasic systems comprising of two immiscible liquid phases intimately mixed and dispersed with each other; in general, emulsions are either of the water-in-oil (w/o) or oil-in-water (o/w) variety.
  • Emulsion formulations have been widely used for oral delivery of therapeutics due to their ease of formulation and efficacy of
  • Liposomes are vesicles that have a membrane formed from a lipophilic material and an aqueous interior that can contain the composition to be delivered. Liposomes can be particularly useful due to their specificity and the duration of action they offer from the standpoint of drug delivery. Liposome compositions can be formed, for example, from phosphatidylcholine, dimyristoyl phosphatidylcholine, dipalmitoyl phosphatidylcholine, dimyristoyl phosphatidylglycerol, or dioleoyl phosphatidylethanolamine.
  • LIPOFECTIN a 1 : 1 (w/w) liposome formulation of the cationic lipid N-[l-(2,3- dioleyloxy)propyl]-n,n,n-trimethylammonium chloride (DOTMA) and dioleoyl
  • DOPE phophotidylethanolamine
  • EFFECTENE a non-liposomal lipid formulation in conjunction with a DNA- condensing enhancer
  • Polypeptides can further encompass any pharmaceutically acceptable salts, esters, or salts of such esters, or any other compound which, upon administration to an animal including a human, is capable of providing (directly or indirectly) the biologically active metabolite or residue thereof. Accordingly, for example, this document provides pharmaceutically acceptable salts of polypeptides, prodrugs and pharmaceutically acceptable salts of such prodrugs, and other bioequivalents.
  • prodrug indicates a therapeutic agent that is prepared in an inactive form and is converted to an active form (i.e., drug) within the body or cells thereof by the action of endogenous enzymes or other chemicals and/or conditions.
  • pharmaceutically acceptable salts refers to physiologically and pharmaceutically acceptable salts of the polypeptides provided herein (i.e., salts that retain the desired biological activity of the parent polypeptide without imparting undesired toxicological effects).
  • pharmaceutically acceptable salts include, but are not limited to, salts formed with cations (e.g., sodium, potassium, calcium, or polyamines such as spermine); acid addition salts formed with inorganic acids (e.g., hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, or nitric acid); and salts formed with organic acids (e.g., acetic acid, citric acid, oxalic acid, palmitic acid, or fumaric acid).
  • cations e.g., sodium, potassium, calcium, or polyamines such as spermine
  • inorganic acids e.g., hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, or nitric acid
  • organic acids e.
  • compositions containing the polypeptides provided herein also can incorporate penetration enhancers that promote the efficient delivery of polypeptides to the skin of animals.
  • Penetration enhancers can enhance the diffusion of both lipophilic and non-lipophilic drugs across cell membranes.
  • Penetration enhancers can be classified as belonging to one of five broad categories, i.e., surfactants (e.g., sodium lauryl sulfate, polyoxyethylene-9-lauryl ether and polyoxyethylene-20-cetyl ether); fatty acids (e.g., oleic acid, lauric acid, myristic acid, palmitic acid, and stearic acid); bile salts (e.g., cholic acid, dehydrocholic acid, and deoxycholic acid); chelating agents (e.g., disodium ethylenediaminetetraacetate, citric acid, and salicylates); and non-chelating non-surfactants (e.g., unsaturated cyclic
  • compositions containing (a) one or more polypeptides and (b) one or more other agents that function by a different mechanism.
  • anti-inflammatory drugs including but not limited to nonsteroidal anti-inflammatory drugs and corticosteroids
  • antiviral drugs including but not limited to ribivirin, vidarabine, acyclovir and ganciclovir
  • non-polypeptide agents e.g., chemotherapeutic agents
  • Such combined compounds can be used together or sequentially.
  • compositions additionally can contain other adjunct components conventionally found in pharmaceutical compositions.
  • the compositions also can include compatible,
  • compositions provided herein can be mixed with auxiliary agents, e.g., lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, colorings, flavorings, and aromatic substances.
  • auxiliary agents e.g., lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, colorings, flavorings, and aromatic substances.
  • auxiliary agents e.g., lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, colorings, flavorings, and aromatic substances.
  • the pharmaceutical formulations which can be presented conveniently in unit dosage form, can be prepared according to conventional techniques well known in the pharmaceutical industry. Such techniques include the step of bringing into association the active ingredients (e.g., the C H 2-C H 3 binding polypeptides provided herein) with the desired pharmaceutical carrier(s) or excipient(s). Typically, the formulations can be prepared by uniformly and bringing the active ingredients into intimate association with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product. Formulations can be sterilized if desired, provided that the method of sterilization does not interfere with the effectiveness of the polypeptide contained in the formulation.
  • compositions described herein can be formulated into any of many possible dosage forms such as, but not limited to, tablets, capsules, liquid syrups, soft gels, suppositories, and enemas.
  • the compositions also can be formulated as suspensions in aqueous, non-aqueous or mixed media.
  • Aqueous suspensions further can contain substances that increase the viscosity of the suspension including, for example, sodium carboxymethylcellulose, sorbitol, and/or dextran.
  • Suspensions also can contain stabilizers.
  • C H 2-C H 3 binding polypeptides can be combined with packaging material and sold as kits for reducing Fc-mediated immune complex formation. Components and methods for producing articles of manufacture are well known.
  • the articles of manufacture may combine one or more of the polypeptides and compounds set out in the above sections.
  • the article of manufacture further may include, for example, buffers or other control reagents for reducing or monitoring reduced immune complex formation. Instructions describing how the polypeptides are effective for reducing Fc-mediated immune complex formation can be included in such kits. Zika Virus
  • Zika virus Zika virus (ZIKV) is a member of the virus family Flaviviridae and the genus Flavivirus and is transmitted by daytime-active Aedes mosquitoes. In humans, the virus causes a typically mild illness known as Zika fever, Zika, or Zika virus disease. Since the 1950s, Zika virus disease has been known to occur within a narrow equatorial belt from Africa to Asia. In 2014, the virus spread eastward across the Pacific Ocean to Central America, the Caribbean, and South America, where the Zika outbreak reached pandemic levels.
  • Zika virus is enveloped and icosahedral and has a non-segmented, single-stranded, positive-sense RNA genome.
  • Zika virus is related to dengue virus, yellow fever virus, Japanese encephalitis virus, and West Nile virus.
  • Zika virus is closely related to the Spondweni virus and is one of the two viruses in the Spondweni virus clade.
  • Zika virus The illness caused by Zika virus is treated by rest, but it cannot yet be treated or prevented by drugs or vaccines. Infection by the Zika virus results in flu-like symptoms and a rash in most infected adults, but the Zika virus causes microcephaly in newborn babies, resulting from mother-to-child transmission. Other neurologic conditions, including Guillain-Barre syndrome (GBS), are likely triggered by Zika virus in a small proportion of infected adults.
  • GBS Guillain-Barre syndrome
  • the Zika virus was first isolated in April 1947 from a rhesus macaque monkey. When the monkey developed a fever, researchers isolated a transmissible agent from its serum. A second isolation of the agent from the mosquito africanus followed at the same site in January 1948. The transmissible agent was first described as Zika virus in 1952. In 1968, Zika virus was isolated for the first time from humans in Nigeria. From 1951 through 1981, evidence of human infection was reported from other African countries including the Central African Republic, Egypt, Gabon, Sierra Leone, Africa, and Kenya, as well as in parts of Asia including India, Indonesia, Malaysia, the Philippines, Thailand, and Vietnam.
  • NS1 nonstructural protein 1
  • Zika virus is transmitted by daytime-active mosquitoes and has been isolated from a number of species in the genus Aedes, including A. aegypti, A. albopictus, A. africanus, A. apicoargenteus, A. furcifer, A. hensilli, A. luteocephalus, and A. vitattus.
  • Zika virus can migrate directly between humans through sexual contact, and it can also cross the placental membrane, affecting an unborn fetus and potentially causing microcephaly in the fetus or miscarriage.
  • Zika virus extends to the range of the mosquito species that transmit it (i.e., its vectors).
  • A. aegypti and albopictus distribution is now the most extensive ever recorded - reaching across all continents.
  • Zika vims Common symptoms of infection with Zika vims include mild headaches, maculopapular rash, fever, malaise, conjunctivitis, and joint pain.
  • symptoms began with a mild headache and progressed to include a maculopapular rash, fever, and back pain. Within two days, the rash started fading, and within three days, the fever resolved and only the rash remained.
  • Zika vims disease has been a relatively mild disease of limited scope, with only one in five persons developing symptoms, and with few known fatalities, but its tme potential as a viral agent of disease is unknown.
  • no vaccine or dmg treatment is available. Symptoms can be treated with rest, fluids, and paracetamol.
  • a link between a transplacental infection of a fetus with Zika vims and microcephaly and brain damage has been confirmed by the WHO and the CDC. Data suggests that newborn babies of mothers who had a Zika vims infection during the first trimester of pregnancy are at an increased risk of microcephaly. Genetic material from Zika vims has been detected in the placenta of a woman after termination of a pregnancy, confirming the vims' s ability to cross the placenta.
  • Zika vims can likely be passed from mother to fetus during any trimester of pregnancy, and limited data indicates that maternal infection with Zika vims in the first trimester might carry a greater risk of fetal microcephaly.
  • other fetal brain abnormalities that have been reported in association with clinical maternal Zika vims infection are
  • GRS Guillain-Barre syndrome
  • other neurologic conditions including, for example, myelitis (inflammation of the spinal cord) or brain abnormalities, may also be complications of Zika vims.
  • myelitis inflammation of the spinal cord
  • brain abnormalities may also be complications of Zika vims.
  • 73 cases of Guillain-Barre syndrome (GBS) and other neurologic conditions occurred in a population of 270,000.
  • the Brazil Ministry of Health reported an increased number of people infected with Zika virus.
  • Circulating immune complexes have been shown to play a role in other viral infections, including dengue fever and dengue hemorrhagic fever (DHF).
  • Immune complexes can form between anti-viral antibodies and viral antigens; between anti-viral antibodies and viral antigens from a different strain of the same virus; or between non-viral antibodies and non-viral antigens.
  • dengue antibodies that bind but do not neutralize viral particles of the second strain of the virus are believed to bring circulating immune complexes into close proximity with the cell surface Fc receptors, which in turn facilitate viral entry into the cells, driving antibody-dependent enhancement (ADE) of dengue virus infection.
  • AD antibody-dependent enhancement
  • an immune complex can form between a Zika virus antibody and a Zika virus antigen.
  • an immune complex can form between a non- Zika virus antibody and a non-Zika virus antigen.
  • an immune complex can form between a non- Zika virus antibody and a non-Zika virus antigen.
  • an immune response to a that different virus or disease could occur before or simultaneously with the Zika virus infection.
  • Such an immune response could result in the formation of heterologous immune complexes.
  • the terms "heterologous immune complex” and “heterologous IC” mean a complex formed between a non-Zika virus antibody and a non-Zika virus antigen.
  • heterologous immune complexes could be formed during Zika virus infection between, for example, antibodies generated during an immune response another virus, e.g., dengue virus, and an antigen from the other virus, e.g., dengue virus antigens.
  • Anti-dengue antibodies have been detected in some patients during Zika virus infection (Duffy et al.(2009), N. Engl. J. Med., 360:2536-2543; CDC MMWR, Jan. 29, 2016, 65(3): 63-67).
  • Such antibodies if cross-reactive to Zika virus, would be expected to have low avidity to the Zika virus and be poorly neutralizing or non-neutralizing.
  • Heterologous immune complexes present during Zika virus infection could also contain self-antigens and antibodies formed from an immune response generated by an autoimmune disease, such as, for example, systemic lupus erythematous (SLE).
  • SLE systemic lupus erythematous
  • nonstructural protein 1 can act as an Fc Receptor.
  • the viral Fc receptor (FcR) activity of Zika virus could, therefore, permit binding of Zika virus to any immune complex including, for example, a heterologous immune complex. Binding to an immune complex that is also bound to a host cell Fc receptor can enhance entry of Zika virus into host cells. Thus, a heterologous immune complex, not just an anti-Zika virus immune complex, could facilitate a host cell-Zika virus interaction. Binding of Zika virus and Zika virus NS1 to non-Zika virus-related immune complexes of peroxidase-rabbit anti-peroxidase IgG (“PAP-IC”) are shown in Example 1.
  • PAP-IC peroxidase-rabbit anti-peroxidase IgG
  • Zika virus and/or Zika virus NS1 may interact with heterologous immune complexes (i.e., ICs formed between a non-Zika virus antibody and a non-Zika virus antigen).
  • Zika virus's ability to interact with heterologous immune complexes may contribute to antibody dependent enhancement (ADE) that accelerates disease progression, increasing Zika virus infectivity and contributing to the pathogenicity of Zika virus infections.
  • ADE antibody dependent enhancement
  • Exemplary diseases that can cause an antibody response resulting in the formation of heterologous immune complexes in humans include, for example, dengue virus (DENV), Toxoplasma gondii, human immunodeficiency virus (HIV), human cytomegalovirus (HCMV), herpes simplex virus (HSV), hepatitis C virus, human papilloma virus (FIPV), Mycobacterium tuberculosis, malaria/ Plasmodium falciparum, and Schistosoma haematobium.
  • FcRn neonatal Fc Receptor
  • immune cells including, for example, monocytes, macrophages, neutrophils, dendritic cells, mast cells, basophils, B cells, NK cells, NKT cells, etc.
  • the neonatal Fc Receptor (FcRn) is expressed on both immune cells including, for example, antigen-presenting cells, monocytes/macrophages, neutrophils, and non-immune cells including, for example, vascular endothelial cells, intestinal epithelial cells, and epithelial cells of the blood-brain barrier.
  • FcRn is also expressed on syncytiotrophoblasts in human placenta which form the epithelial covering of the highly vascular embryonic placental villi. FcRn is occasionally or weakly expressed on fetal vessel endothelium in placental tissues. Although FcRn has not been reported to be expressed on Hofbauer cells, mononuclear phagocytes found in the placental, FcyRI is expressed on Hofbauer cells (Saji et al. (1999) Rev. Reprod. 4(2):81-9).
  • the presence of the Fc-receptor expressing cells in the placenta present the possibility that Fc- mediated transcytosis of immune complexes can increase vertical transmission from mother to fetus.
  • the immune complexes can include, for example, an anti-Zika virus IgG and Zika virus, a cross reactive IgG and Zika virus, and/or a heterologous IC (a complex formed between a non- Zika virus antibody and a non-Zika virus antigen).
  • Zika virus's Fc-binding capacity permits it to bind to the IC, allowing Zika virus to be transcytosed across the placental barrier along with the IC.
  • Endothelial cells at the blood brain barrier also express FcRn, and FcRn-mediated transcytosis of Zika virions into the CNS may contribute to microcephaly and/or Guillain-Barre syndrome (GBS).
  • Endothelial cells of the testes can also express FcRn, and FcRn-mediated transcytosis of Zika virions into the testes may contribute to sexual transmission of Zika virus.
  • CMV is the prototypical virus able to infect the placenta and cause vertical transmission from mother to fetus.
  • Neonatal Fc receptor (FcRn) expressed in early-gestation placenta binds IgG- CMV virion complexes, protecting them from degradation, and FcRn transcytoses virions in IgG-CMV virion complexes from the mother to the fetus.
  • FcRn Neonatal Fc receptor expressed in early-gestation placenta binds IgG- CMV virion complexes, protecting them from degradation, and FcRn transcytoses virions in IgG-CMV virion complexes from the mother to the fetus.
  • Zika virus may use a similar pathway as CMV during vertical transmission of the virus to a fetus.
  • Polypeptides that bind to the C H 2-C H 3 cleft of an immunoglobulin molecule and inhibit Fc- mediated immune complex formation have been suggested as useful for treatment of ADE in dengue fever and dengue hemorrhagic fever. (U.S. Patent No. 8,815,813). Due to the unexpected finding that Zika virus and/or Zika virus NSl act as an Fc Receptor, such polypeptides may also abrogate interactions between Zika virus and a heterologous immune complex that includes a non-Zika virus antibody and a non-Zika virus antigen.
  • polypeptides could also abrogate interactions between immune complexes and FcRn where the immune complex includes Zika virus as an antigen bound to the antibody (e.g., IgG). Due to the role of FcRn in maternal -fetus viral transmission, polypeptides that inhibit Fc-mediated immune complex formation could be effective vertical transmission deterrents when applied in the context of Zika virus infections.
  • C H 2-C H 3 binding polypeptides can be used in in vitro assays of Fc-mediated immune complex (IC) formation. Such methods are useful to, for example, evaluate the ability of a C H 2-C H 3 cleft- binding polypeptide to block Fc-mediated immune complex formation.
  • In vitro methods can involve, for example, contacting an immunoglobulin molecule (e.g., an antigen bound immunoglobulin molecule) with an effector molecule (e.g., mClq, sClq, FcRs, FcRn, DC-SIGN, or an antibody) in the presence and absence of a polypeptide as provided herein, and determining the level of IC formation in each sample.
  • an immunoglobulin molecule e.g., an antigen bound immunoglobulin molecule
  • an effector molecule e.g., mClq, sClq, FcRs, FcRn, DC-SIGN, or an antibody
  • Levels of IC formation can be evaluated by, for example, polyacrylamide gel electrophoresis with Coomassie blue or silver staining, or by co- immunoprecipitation. Such methods are known to those of ordinary skill in the art, and can be used to test the ability of a candidate polypeptide or compound to inhibit IC formation associated with an infectious disease, for example.
  • Methods provided herein also can be used to inhibit immune complex formation in a subject, and to treat an infectious viral disease or condition in a subject by inhibiting immune complex formation.
  • the immune complex formation can include complement or Fc-mediated immune complex formation Such methods can include, for example, administering any of the
  • the Zika virus can be African lineage Zika virus or the Asian lineage Zika virus.
  • the Zika virus can be Zika virus strain MR 766.
  • the immune complex can include a Zika virus antibody and a Zika virus antigen. In some embodiments, the immune complex can include a non-Zika virus antibody and a non-Zika virus antigen. In some embodiments, the immune complex can include a non-Zika virus antibody and a Zika virus antigen. In some embodiments, an immune complex can include an Fc-mediated immune complex. In some embodiments, the immune complex can include an FcyR or an FcRn. In some embodiments, the immune complex can include mClq, or sClq. In some embodiments, the immune complex can include and/or can interact with DC-SIGN.
  • the immune complex can include a Zika virus and/or a Zika virus protein including, for example nonstructural protein 1 (NS1).
  • NS1 nonstructural protein 1
  • an immune complex includes an Fc-mediated immune complex and a Zika virus and/or a Zika virus protein
  • the Zika virus and/or the Zika virus protein can act as an Fc Receptor.
  • the subject may also have or be at risk of having another viral disease alone or in combination with Zika virus including, for example, dengue virus (DENV), Toxoplasma gondii, human immunodeficiency vims (HIV), human cytomegalovirus (HCMV), herpes simplex virus (HSV), hepatitis C virus, human papilloma virus (HPV), Mycobacterium tuberculosis,
  • DECV dengue virus
  • Toxoplasma gondii human immunodeficiency vims
  • HMV human cytomegalovirus
  • HSV herpes simplex virus
  • HPV human papilloma virus
  • Mycobacterium tuberculosis Zika virus including, for example, dengue virus (DENV), Toxoplasma gondii, human immunodeficiency vims (HIV), human cytomegalovirus (HCMV), herpes simplex virus (HSV), hepatitis C virus, human papillom
  • an antibody response against the second viral disease can result in the formation of heterologous immune complexes.
  • the subject can be a mammal. In some embodiments, the subject can be a human or a non-human primate. In some embodiments, the subject may be pregnant. In some embodiments, the subject has been diagnosed as having or is suspected of having Zika-associated Guillain-Barre syndrome (GBS).
  • GGS Zika-associated Guillain-Barre syndrome
  • a method can include administering to an individual a composition containing a polypeptide that includes the amino acid sequence Cys-Ala-Trp-His-Leu-Gly-Glu-Leu-Val-Trp- Cys-Thr (SEQ ID NO: 10).
  • a method can include administering to a subject a polypeptide that contains the amino acid sequence Asp-Cys-Ala-T ⁇ -His-Leu-Gly-Glu-Leu-Val- Trp-Cys-Thr (SEQ ID NO:2), Xaa-Pro-Pro-Asp-Cys-Ala-Trp-His-Leu-Gly-Glu-Leu-Val-Trp- Cys-Thr (SEQ ID NO: 19; where Xaa is any amino acid), or Ala-Pro-Pro- Asp-Cys-Ala-Trp-His- Leu-Gly-Glu-Leu-Val-Trp-Cys-Thr (SEQ ID NO:20).
  • a polypeptide when the viral disease is a Zika virus infection, a polypeptide can be administered to inhibit or prevent maternal-fetus Zika virus transmission.
  • immune complex formation can include formation of an immune complex capable of being transcytosed.
  • the transcytosis can include FcRn-mediated transcytosis.
  • the transcytosis can include at least one of placental transcytosis, blood brain barrier transcytosis, retinal transcytosis, epithelial cell transcytosis, endothelial cell transcytosis, and testicular transcytosis (including testicular endothelial cell transcytosis).
  • the immune complex is preferably an Fc-mediated immune complex.
  • immune complex formation can enhance transcytosis of Zika virus by bridging infectious Zika virions and FcRn.
  • transcytosis of an immune complex IC
  • blocking Zika virus-binding to an IC or blocking FcR receptor binding to an IC with a polypeptide could prevent vertical transmission from the mother to the fetus.
  • blocking Zika virus-binding to an IC or FcRn binding to an IC can inhibit and/or abrogate transcytosis of an IC including Zika virus.
  • the infection of placental macrophages (Hofbauer cells) and/or dendritic cells via FcyRs may involve ADE of IgG complexes bound to the viral FcR of Zika virus and/or Zika virus NS 1. Since a C H 2-C H 3 binding polypeptide can inhibit IC IgG hexamers from binding to Clq, FcyRI, FcyRIIa, and Fcylll, a C H 2-C H 3 binding polypeptide can also inhibit IC IgG hexamers from binding to the viral Fc receptor of Zika virus, preventing the infection of the Hofbauer cells and DC in the placenta by Zika virus.
  • the polypeptide may inhibit the binding to an FcR on a placental cell of an immune complex that includes an antibody and an antigen (e.g., IgG IC).
  • the FcR on a placental cell can be FcRn.
  • the immune complex that includes an antibody and an antigen can include Zika virus as an antigen.
  • the immune complex that includes an antibody and an antigen can be a heterologous immune complex.
  • the immune complex that includes an antibody and an antigen can be bound to Zika virus via a viral Fc receptor
  • transcytosis e.g., of an infectious Zika virion bound to an IgG immune complex
  • GSS Zika virus-associated Guillain-Barre syndrome
  • blocking Zika virus-binding to an IC or blocking FcR binding to an IC with a polypeptide could ameliorate the neurological symptoms associated with Guillain-Barre syndrome.
  • testicular transcytosis of an IC contributes to sexual transmission of Zika virus
  • blocking Zika virus-binding to an IC or blocking FcR binding to an IC with a polypeptide could ameliorate the sexual transmission of Zika virus.
  • a polypeptide when the viral disease is Zika virus infection, can be administered to inhibit immune complex formation that is associated with Zika virus infection, that is associated with antibody dependent enhancement (ADE) of Zika virus infection, that contributes to the enhancement of a Zika virus infection, or that contributes to the transmission of a Zika virus infection.
  • ADE antibody dependent enhancement
  • ADE can include complement-mediated antibody-dependent enhancement (C-ADE) and/or ADE mediated by DC-SIGN.
  • the immune complex formation is preferably Fc-mediated immune complex formation.
  • the polypeptide can be administered to inhibit immune complex formation between DC-SIGN and an immune complex that includes an antibody and an antigen.
  • the immune complex that includes an antibody and an antigen can include Zika virus as an antigen. In some embodiments the immune complex that includes an antibody and an antigen can be a heterologous immune complex. In some embodiments the immune complex that includes an antibody and an antigen can be bound to Zika virus via a viral Fc receptor
  • the polypeptide can, for example, result in clinical or histological improvement of a Zika virus infection, result in an improvement or delay of onset of one or more of the histological characteristics of a Zika virus infection, result in a decrease maternal-fetal transmission of a Zika virus infection, result in a decrease the ADE of a Zika virus infection, and/or result in a decrease in symptoms of a neurological disorder associated with a Zika virus infection.
  • the neurological disorder can include Guillain-Barre syndrome (GBS) or microcephaly.
  • the methods described herein can include monitoring a subject for one or more clinical, histopathological or molecular characteristics of Zika virus infection and/or for one or more clinical, histopathological or molecular characteristics of a neurological disorder.
  • the neurological disorder can include Guillain-Barre syndrome (GBS).
  • GRS Guillain-Barre syndrome
  • the monitoring can occur before and/or after administration of the polypeptide.
  • the polypeptide can inhibit binding of a heterologous IC to an FcyR; inhibit formation of IC that contributes to immunopathogenesis of the ADE of Zika virus infections; inhibit formation of IC that contributes to maternal -fetal transmission of a Zika virus infection; inhibit binding of Zika virus virion to an IC; inhibit binding of a Zika virus protein (e.g., NS1) to an IC; inhibit binding of Zika virus and/or TNF-a to an IC; inhibit binding of a Zika virus-IgG (or a cross reactive anti-DENV Zika virus-IgG) IC to Fcyl, a Fcylla HI 31 allele, a Fcylla R131 allele, FcyRIIb, FcyRIIc, FcyRIIIa, or Fcylllb, FcRn, and/or DC-SIGN; and/or inhibit binding of Zika virus-IgG (or IgG to other viruses that are
  • a polypeptide can be administered to a subject that has been diagnosed as having a Zika virus infection, a subject suspected of having a Zika virus infection, a subject exhibiting symptoms of a Zika virus infection, a subject that is at risk of contracting a Zika virus infection, and/or a subject that has been exposed to a Zika virus.
  • the subject may be pregnant, suspected of being pregnant, or trying to become pregnant.
  • the subject and/or fetus may also be monitored indications of vertical transmission of Zika virus from the subject to the fetus.
  • a polypeptide can be administered to minimize the risk of a subject developing a Zika virus infection.
  • a polypeptide could be administered to a subject in a hospitalized population wherein one or more other subjects in the same hospitalized population has a Zika virus infection, is suspected of having a Zika virus infection, and/or is exhibiting symptoms of a Zika virus infection.
  • a polypeptide can be administered to a subject that has been diagnosed as having or is suspected of having a coinfection with a co-infective pathogen and has a Zika virus infection, is suspected of having a Zika virus infection, is exhibiting symptoms of a Zika virus infection, is at risk of contracting a Zika virus infection, and/or has been exposed to a Zika virus.
  • the co-infective pathogen may include, for example, dengue virus (DENV), Toxoplasma gondii, human immunodeficiency virus (HIV), human cytomegalovirus (HCMV or CMV), herpes simplex virus (HSV), hepatitis C virus, human papilloma virus (HPV), Mycobacterium tuberculosis, malaria/ Plasmodium falciparum, Schistosoma haematobium, and/or other disease associated with antibody production and/or immune complex formation.
  • DECV dengue virus
  • HMV human immunodeficiency virus
  • HCMV or CMV human cytomegalovirus
  • HSV herpes simplex virus
  • HPV hepatitis C virus
  • Mycobacterium tuberculosis malaria/ Plasmodium falciparum
  • Schistosoma haematobium and/or other disease associated with antibody production and/or immune complex formation.
  • Administration of a polypeptide may be particularly helpful in co-infected individuals or individuals at risk of being co-infected because of the potential for non-Zika virus-related immune complexes to allow Zika virus association with and entry into host cells and/or placental cells.
  • a polypeptide can be administered to a subject that has previously been diagnosed as having or is suspected of having previously had an infection with a disease associated with antibody production and/or immune complex formation and that has a Zika virus infection, is suspected of having a Zika virus infection, is exhibiting symptoms of a Zika virus infection, is at risk of contracting a Zika virus infection, and/or has been exposed to a Zika virus.
  • Such administration may be helpful in individuals that have previously had an infection with a disease associated with antibody production and/or immune complex formation because of the potential for non-Zika virus-related immune complexes to allow Zika virus association with and entry into host cells and/or placental cells.
  • a polypeptide can be administered as soon as a subject is diagnosed with a Zika virus infection, is suspected of having a Zika virus infection, is exhibiting symptoms of a Zika virus infection, is determined to be at risk of contracting a Zika virus infection, and/or has been exposed to a Zika virus.
  • a polypeptide can be administered to a pregnant subject who is at risk of Zika virus infection or who has a Zika virus infection.
  • a polypeptide can be administered to a subject who has or is suspected of having pregnancy-related complications resulting from a Zika virus infection.
  • a polypeptide can be administered to a subject having a replicating Zika virus.
  • a polypeptide can be administered to a subject experiencing symptoms of a neurological disorder.
  • Administration of a polypeptide to a subject who is exhibiting symptoms of a neurological disorder during or even after a Zika virus infection may still alleviate the symptoms and detrimental effects of Zika virus infection because of the probable link between Zika and neurological diseases, including, but not limited to, Guillain-Barre syndrome (GBS) and microcephaly.
  • GBS Guillain-Barre syndrome
  • microcephaly including, but not limited to, Guillain-Barre syndrome (GBS) and microcephaly.
  • a polypeptide can be administered to a subject that has been diagnosed as having or is suspected of having an autoimmune disease and as having or is suspected of having a Zika virus infection, is suspected of having a Zika virus infection, is exhibiting symptoms of a Zika virus infection, is at risk of contracting a Zika virus infection, and/or has been exposed to Zika virus.
  • the autoimmune disease may include, for example, Guillain-Barre syndrome (GBS), systemic lupus erythematosus, cryoglobulinemia, rheumatoid arthritis, scleroderma, Sjogren's syndrome, and/or another autoimmune disease associated with antibody production and/or immune complex formation.
  • a polypeptide can be administered to a subject in combination with another suitable therapy.
  • a polypeptide can be administered to a subject in combination with a composition comprising a recombinant human protein or a therapeutic antibody.
  • a therapeutic antibody can include a monoclonal antibody to FcyRIIb including, for example, SuppreMol SM201 or Xencor XmAb5871.
  • composition comprising monoclonal antibody to FcyRIIb can be administered to a subject before, during, or after administration of a composition comprising the polypeptide.
  • a recombinant human protein such as soluble FcyRIIb including, for example, SuppreMol SM101 can be administered to a subject before, during, or after administration of a composition comprising the polypeptide.
  • a therapeutic antibody can include a monoclonal antibody to FcyRIIb including, for example, SuppreMol SM201 or Xencor XmAb5871 administered during or after a Zika virus infection.
  • the composition comprising monoclonal antibody to FcyRIIb can be administered to a subject before, during, or after a Zika virus infection.
  • a recombinant human protein, such as soluble FcyRIIb including, for example, SuppreMol SMI 01 can be administered to a subject before, during, or after a Zika virus infection.
  • a therapeutic antibody can include an antibody to FcRn including, for example, a monoclonal antibody to FcRn. (Christianson et al. (2012) MAbs, 4(2):208-16).
  • an antibody to FcRn can abrogate and/or inhibit FcRn-mediated
  • an antibody to FcRn can abrogate and/or inhibit FcRn binding to an immune complex.
  • the composition comprising antibody to FcRn can be administered to a subject before, during, or after administration of a composition comprising the polypeptide.
  • a therapeutic antibody can include an antibody to DC-SIGN including, for example, a monoclonal antibody to DC-SIGN.
  • an antibody to DC- SIGN can abrogate and/or inhibit DC-SIGN binding to an immune complex.
  • the composition comprising antibody to DC-SIGN can be administered to a subject before, during, or after administration of a composition comprising the polypeptide.
  • a polypeptide can be administered to a subject in combination with a composition including an antiretroviral including, for example, oseltamivir, zanamivir, peramivir, etc.
  • the composition comprising the antiretroviral can be administered to a subject before, during, or after administration of a composition comprising the polypeptide.
  • a polypeptide can be administered to a subject in combination with an composition including a drug approved for treating hepatitis including sofosbuvir and/or ledipasvir.
  • the composition comprising the drug approved for treating hepatitis can be administered to a subject before, during, or after administration of a composition comprising the polypeptide.
  • a polypeptide can be administered to a subject in combination with a treatment for Guillain-Barre syndrome (GBS).
  • GBS Guillain-Barre syndrome
  • a polypeptide can be administered to a subject in combination with plasmapheresis.
  • a polypeptide can be administered to a subject in combination with an intravenous
  • immunoglobulin (IVIg).
  • In vitro assays involving enzyme-linked immunosorbent assay (ELISA) can be used to demonstrate competitive inhibition of immune complexed IgG Fc binding to immune mediating factors such as Fc receptors (FcRs), (e.g., FcyRI, Fcylla, FcyRIIb/c, FcyRIIIa/b), FcRn, mClq, and sClq by the polypeptides and compounds described herein.
  • FcRs Fc receptors
  • FcRs Fc receptors
  • FcRs Fc receptors
  • FcRs Fc receptors
  • Standardized reagents and ELISA kits are useful to reduce costs and increase the reproducibility of the experiments.
  • SEQ ID NO:20 is a classic noncompetitive allosteric inhibitor designed to bind to the Fc region of IgG immune complexes (IC) and prevents IgG-IC from binding to Fc receptors (FcRs)
  • IC IgG immune complexes
  • FcRs Fc receptors
  • ELISA enzyme-linked immunosorbent assay
  • an antigen is immunoadsorbed onto a plastic microwell. After suitable blocking and washing steps, a primary antibody with specificity directed toward the antigen is added to the microwell.
  • a secondary antibody that is directed toward the primary antibody and conjugated to an enzyme marker, such as horseradish peroxidase (HRP) is added to the microwell.
  • an enzyme marker such as horseradish peroxidase (HRP)
  • HRP horseradish peroxidase
  • the appropriate enzyme substrate such as 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) in the case of HRP, is added.
  • ABTS 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid
  • the conjugated enzyme catalyzes a colorimetric chemical reaction with the substrate, which is read with a microplate reader or spectrophotometer.
  • a titer of the primary antibody (the variable) is established.
  • the primary antibody binds to the antigen through its complementarily determining regions (CDR) located on the Fab arms.
  • CDR complementarily determining regions
  • HRP is conjugated to the Fc region of the secondary antibody, direct Fc binding is very limited or abrogated.
  • a "reverse" ELISA technique is used to assess binding of the Fc receptor ligands that bind to IgG-IC.
  • the enzyme e.g., HRP
  • HRP peroxidase-rabbit anti-peroxidase IgG
  • SEQ ID NO:20 was designed to bind to human IgG-IC and since rabbit IgG is virtually identical to human IgG at the binding site for SEQ ID NO:20, rabbit IgG-IC can be used as a substitute for human IgG-IC, which are not commercially available.
  • HRP serves both as the antigen and the enzyme marker but does not block the Fc region.
  • a Fc binding ligand or FcR (e.g., human FcRIIa or Clq) is bound to microwell plates.
  • PAP IC complexes bind to the immobilized ligand and the reaction between HRP and its substrate produces a signal ("negative control").
  • This signal is reduced by the pre-incubation of inhibitors (e.g., FcRIIa, Clq or rheumatoid factor) or peptides such as SEQ ID NO:20 that inhibits PAP IC from binding to the immobilized FcR ligand.
  • inhibitors e.g., FcRIIa, Clq or rheumatoid factor
  • SEQ ID NO:20 that inhibits PAP IC from binding to the immobilized FcR ligand.
  • less color development is indicative of greater inhibition. See, e.g., U.S. Patent Nos. 6,916,904, 7,714,104, 8,362,202 and 8,815,813.
  • Preparing the FcR coated microwell plates 100 microliters of a 0.05 microgram per microliter ⁇ g ⁇ L) solution of human FcRIIA (rhFcRIIa) or Clq is used to coat each well that is to be used of a standard microwell plate that has 96 0.32 square centimeter (cm 2 ) wells.
  • the rhFcRIIa is prepared by mixing 0.1 milliliter (mL) of 100 ⁇ g/200 [iL rhFcRIIa (catalogue number sc- 174810, Santa Cruz Biotechnology, Dallas, TX, or catalogue number 1330-CD-
  • the plates are blocked from any further non-specific protein adsorption by adding 100 ⁇ of 1 milligram per milliliter (mg/mL) bovine serum albumin (BSA), Cohn fraction V (catalogue number A5611, Sigma- Aldrich, St. Louis, MO) to each of the wells.
  • BSA bovine serum albumin
  • Cohn fraction V catalogue number A5611, Sigma- Aldrich, St. Louis, MO
  • the 1 mg/mL BSA solution is prepared by dissolving BSA in IX PBS and then filter sterilizing. After allowing the plate to incubate for 1 hour at 4°C, the BSA solution is manually removed and each well is washed four more times with 100 ⁇ L of the 1 mg/mL BSA solution with no additional time between washes. One final wash with 100 ⁇ L of IX PBS is then done to remove any residual BSA. If the plate is sealed it can be used for up to several days later.
  • IC Forming the PAP immune complex (IC). 10 milligrams (mg) of HRP (catalogue number P6782, Sigma-Aldrich, St. Louis, MO) is dissolved in 2 mL of IX PBS. Lyophilized rabbit anti- peroxidase antibody (catalogue number P7899, Sigma-Aldrich, St. Louis, MO) is dissolved in 2.1 mL of ice cold IX PBS and filtered using a 0.22 micrometer ( ⁇ ) pore size 13 millimeter (mm) PVDF syringe filter and a 3 mL sterile syringe. The resuspended rabbit anti-peroxidase antibody is stored at 4°C.
  • HRP catalogue number P6782, Sigma-Aldrich, St. Louis, MO
  • Lyophilized rabbit anti- peroxidase antibody catalog number P7899, Sigma-Aldrich, St. Louis, MO
  • the rabbit antibody must be handed using sterile techniques to avoid bacterial contamination since the P7899 antibody does not contain a preservative agent.
  • the resuspended antibody can be used up to one year as long as no contamination occurs as judged by an increase in turbidity.
  • 20 ⁇ L of the resuspended rabbit anti- peroxidase antibody is added to 950 ⁇ of IX PBS, followed by the addition of 50 ⁇ of the 5 mg/mL HRP solution.
  • the PAP complex must be used immediately. It cannot be stored, because even with antigen excess, larger immune complexes will be formed upon storage.
  • each inhibitor/PAP complex mixture is added to a well of the coated microwell plate as well as the control PAP complex without the added inhibitor. After allowing the plate to incubate for 1 hour, the inhibitor/PAP complex mixture is manually removed and each well is washed four more times with 100 ⁇ of IX PBS with no additional time between washes.
  • PAP immune complexes were formed by mixing 2 of rabbit anti-peroxidase (P7899, Sigma-Aldrich, St. Louis, MO) with 50 [iL of peroxidase (P6782, Sigma-Aldrich, St. Louis, MO) in 1 mL distilled water. PAP (100 ⁇ iL) were pre-incubated with 100 [iL of human Clq (Quidel Corp., San Diego, CA) or 100 [iL of peptide (see Table 1) for 1 hour. Like FcR, Clq is known to bind to the Fc region on IgG.
  • the ability of PAP immune complexes to bind to the immobilized Clq in the presence or absence of inhibitors was determined.
  • the Clq/PAP mixture served as a positive control; PAP immune complexes bind to soluble Clq, and the premixed Clq/PAP immune complexes are therefore not expected to bind to the Clq-containing plate, resulting in low signal levels, similar to what would be observed in the presence of an inhibitor.
  • the PAP immune complex alone serves as the negative control; in the absence of inhibitors, the PAP immune complex is expected to bind to the Clq-containing plate, resulting in high signal levels.
  • PAP/peptide mixtures were added to the FcyR coated plates and incubated for one hour. After washing, plates were incubated with ABTS substrate (Quidel Corp., San Diego, CA) for 15 minutes and read at 405 nm. Results are shown in Table 2.
  • Peptide APPDCAWHLGELVWCT (SEQ ID NO:20) appeared to result in the greatest overall inhibition of FcR binding to PAP, followed by peptide DCAWHLGELVWCT (SEQ ID NO:2). Experiments with SEQ ID NO:20 were repeated. COSTAR microtiter plates were coated with 1 : 10 dilutions of highly purified Fcylla (H is 131 allele aka HI 61), Fcyllb and Fcylllb and incubated for 24 hours. The plates were washed and then blocked with 10 mg/mL BSA blocking solution for 24 hours. PAP immune complexes were formed as described in Example 2. PAP (100 ⁇ ) were pre-incubated with 100 ⁇ of peptide for one hour. PAP/peptide mixtures were added to the FcyR coated plates and incubated for one hour. After washing, plates were incubated with ABTS substrate for 15 minutes and read at 405 nm. Results are shown in Table 3.
  • SEQ ID NO:20 inhibited binding of all three major classes of Fc receptor (Fcyl,
  • Zika virus as a viral FcR Zika virus contains a viral FcR, and ICI406 (SEQ ID NO: 20) can prevent the binding of IC to Zika virus
  • ICI406 SEQ ID NO: 20
  • the reverse ELISA assay was used. Instead of binding a known FcR to the plate, such as human FcRIIa or Clq, Zika Virus (live Zika virus, # 0810092CF, ZeptoMetrix Corporation, Buffalo, NY) was used. 100 ⁇ .
  • Zika virus contains a viral FcR that can bind IC (and, in particular, peroxidase-rabbit-anti-peroxidase IgG) and that SEQ ID NO:20 can effectively prevent the binding of an IC to the Zika virus.
  • Zika NSl protein as a viral FcR Zika virus NSl protein is a viral FcR, and ICI406 (SEQ ID NO: 20) can prevent the binding of an IC to Zika virus NSl protein
  • Zika virus nonstructural protein 1 was investigated as a candidate viral FcR.
  • the reverse ELISA assay was used to test whether the Zika virus NSl protein was a viral FcR and whether SEQ ID NO:20 could inhibit binding of an IC to the NSl protein of the Zika virus.
  • Recombinant Zika virus NSl from Kenya strain MR766 ZIKV-NSl-100, Native Antigen Company, Oxfordshire, UK
  • the Zika NSl core protein was used at a concentration of 10 or 3.0 ⁇ g/mL.
  • the standard reverse ELISA protocol was implemented with the inhibitor SEQ ID NO:20 used at a concentration of 10 mg/mL, 3 mg/mL, and 1 mg/mL; OD was read at 405 nm. Results are shown in Table 5.
  • Zika virus NSl protein (ZIKV NSl) is a viral FcR that can bind an IC and that SEQ ID NO:20 can effectively prevent the binding of an IC to the Zika virus NSl protein.
  • FcRn (#8639-FC- 050, R&D Systems, Minneapolis, MN) was immobilized on microwell plates. PAP immune complexes were formed by mixing 2 ⁇ L. of rabbit anti-peroxidase (P7899, Sigma-Aldrich, St. Louis, MO) with 50 uL of peroxidase (P6782, Sigma-Aldrich, St. Louis, MO) in 1 mL distilled water. Like FcR, FcRn is known to bind to the Fc region on IgG. The ability of PAP immune complexes to bind to the immobilized FcRn was determined as described above. OD was read at 405 nm. Results are shown in Table 6.
  • DC-SIGN is known to bind to sialyled IgG Fc (Sondermann et al. (2013) PNAS, 110(24):9868-72).
  • Non-sialyled IgG has not been reported to bind to DC-SIGN.
  • the ability of "normal" (non-sialyled) IgG PAP immune complexes to bind to immobilized DC-SIGN was determined. Unexpectedly, non-sialyled IgG PAP immune complexes bind to DC-SIGN.
  • IgG hexamers form a ring structure that allows high affinity binding by Clq (Diebolder et al. (2014) Science, 343(6176): 1260-3).
  • SEQ ID NO:20 (ICI406) inhibits Clq binding to ICs and is believed to disrupt the IgG Fc-Fc interactions required to form an IgG hexamer.
  • SEQ ID NO:20 (ICI406) inhibits Zika NSl binding to an IC, an in silco model of Zika NSl binding to IgG was created .
  • FIG. 1 shows the results of in silco modeling, indicating that it is possible that a Zika NSl hexamer can bind an IgG hexamer.
  • SEQ ID NO:20 (ICI406) inhibits Zika NS1 hexamer binding to IgG (hexamer) by disrupting the Fc-Fc interactions required to form an IgG hexamer.
  • the idea that Zika NS1 is mimicking Clq is consistent with the experimental data disclosed herein, but was completely unexpected.
  • ZIKV Zika virus
  • the plates were washed 5 times with ELISA wash solution (Quidel Corp., San Diego, CA), blocked with highly purified BSA for 1 hour, and washed 5 times.
  • ELISA wash solution Quidel Corp., San Diego, CA
  • Rabbit anti-ZIKV IC are formed by combining ( ⁇ ) whole inactivated Zika virus (Zika Virus Culture Fluid, # 0810092CF, ZeptoMetrix Corporation, Buffalo, NY) with an antibody that reacts with Zika, rabbit IgG Anti-Flavivirus group antigen antibody (clone number D1-4G2-4- 15 (4G2), catalogue number Ab00230-23.0, Absolute Antibody Ltd, Oxford, United Kingdom.
  • the antibody and the Zika virus are pre-incubated with and without SEQ ID NO:20 ( ⁇ at 10 mg/ml, 3.3 mg/ml or 1.1 mg/ml) for 1 hour.
  • an anti-Zika HRP conjugate can be used instead of the Mouse Anti-FlaviVirus group antigen antibody and HRP- conjugated anti-mouse secondary antibody, described below.
  • HRP conjugate conjugate number A- 10685, ThermoFisher Scientific, Minneapolis, MN
  • 100 ⁇ _ of ABTS is added to each microwell; after 30 minutes incubation, the plates are read at 405 nm.
  • ICI406 is expected to inhibit the binding of Zika-IgG immune complexes to FcyRIIa.
  • Immune complexes in vitro and in vivo, can bridge Zika viral FcR to cellular FcyR, primarily recombinant human FcyRIIa, and allow infection (either cis or trans) of immunocompetent cells, such as macrophages.
  • the Zika viral FcR allows an alternate means to drive ADE.
  • SEQ ID NO:20 (ICI406) can disrupt ADE by disruption of IgG- Fc to Fc interaction and inhibition of binding of FcRIIa/CD32a to IgG Fc and/or binding of ZIKV viral FcR to IgG Fc.
  • a three-dimensional culture system that recapitulates placental syncytiotrophoblast development and microbial resistance is used to test whether ICI406 can inhibit FcRn mediated transcytosis of ZIKV in syncytiotrophoblast cells.
  • ZIKV virions are incubated with enhancing antibodies obtained in the manner described by Paul et al. (bioRxiv Online 4-25-16, Dengue Virus Antibodies Enhance Zika Virus Infection) to form infectious ZIKV IC.
  • ICI406 is added at a concentration of 0 mg/mL or lOmg/mL to the infectious ZIKV IC for one hour at 37°C, before the ICs are added to 3-D cell cultures prepared as described in McConkey et al. (2016) Sci. Adv., 2:el501462. Immunochemistry is used to demonstrate translocation of infectious ZIKV and RT-PCR is used to measure infection of the trophoblasts. The presence of ICI406 is expected to abrogate infection of the trophoblasts.
  • the rabbits are monitored for clinical and histological evidence of Axonal Guillain-Barre Syndrome.
  • Axonal Guillain-Barre Syndrome is expected to be induced in immunized rabbits, and abrogation of clinical and/or histological features of Guillain-Barre Syndrome are expected to be abrogated in rabbits treated with ICI406.

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Abstract

Cette invention concerne des polypeptides et autres composés qui se lient spécifiquement à la fissure CH2-CH3 d'une molécule d'immunoglobuline, ainsi que des méthodes d'utilisation de ces polypeptides et composés pour inhiber la formation d'un complexe immun à médiation Fc dans l'infection virale. Par exemple, des polypeptides et d'autres composés peuvent être utilisés pour inhiber la formation de complexes immuns chez un sujet qui a été diagnostiqué comme atteint ou suspecté d'être atteint d'une infection par le virus Zika.
PCT/US2017/029918 2016-04-27 2017-04-27 Méthodes de traitement d'infections par le virus zika et de complications associées WO2017189891A1 (fr)

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2018208231A1 (fr) * 2017-05-11 2018-11-15 National University Of Singapore Prévention ciblée de la transmission verticale d'une infection entre une mère et son fœtus
US11673955B2 (en) 2017-05-11 2023-06-13 National University Of Singapore Targeted prevention of maternal to foetal vertical transmission of infection
WO2019083904A1 (fr) * 2017-10-23 2019-05-02 Chan Zuckerberg Biohub, Inc. Mesure de glycanes fc d'igg afucosylés et procédés de traitement associés
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CN111683959A (zh) * 2017-11-09 2020-09-18 巴斯德研究院 包含非结构蛋白的寨卡病毒嵌合多表位及其在免疫原性组合物中的用途
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