Nanobodies to henipavirus and uses thereof

Abstract

Disclosed are compounds, compositions, and methods involving nanobodies that bind Henipavirus. In some forms, the nanobodies can neutralize Henipavirus. The disclosed nanobodies are generally useful for binding to and affecting Henipavirus, such as Hendra virus and Nipah virus. The disclosed nanobodies are useful for treating Henipavirus infections.

Description

[0001] NANOBODIES TO HENIPA VIRUS AND USES THEREOF

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] This application claims the benefit of and priority to U.S. Provisional Application No.

[0004] 63 / 662,948 filed on June 21, 2024, the contents of which is incorporated herein in its entirety.

[0005] REFERENCE TO THE SEQUENCE LISTING

[0006] The Sequence Listing XML submitted as a file named “GSURF_2024-018- 02PCT_ST26.xml”, created on June 23, 2025, and having a size of 36,488 bytes is hereby incorporated by reference pursuant to 37 C.F.R. § 1.834(c)(1).

[0007] FIELD OF THE INVENTION

[0008] The disclosed invention is generally in the field of Henipavirus and specifically in the area of treatment of Henipavirus infection.

[0009] BACKGROUND OF THE INVENTION

[0010] Henipavirus is a in the Paramyxoviridae family. They are zoonotic pathogens with fatality rates of 50-100% for the most prominent forms (Nepah virus and Hendra virus). New strains continue to emerge. There are no FDA-approved therapeutics or vaccines for Henipavirus infections. The World Health Organization has made Henipavirus priority pathogens of epidemic and pandemic potential and found that there is an urgent need for research and development.

[0011] Any discussion of documents, acts, materials, devices, articles or the like which has been included in the present specification is not to be taken as an admission that any or all of these matters form part of the prior art base or were common general knowledge in the field relevant to the present disclosure as it existed before the priority date of each claim of this application.

[0012] Throughout this specification the word “comprise,” or variations such as “comprises” or “comprising,” will be understood to imply the inclusion of a stated element, integer or step, or group of elements, integers or steps, but not the exclusion of any other element, integer or step, or group of elements, integers or steps.

[0013] BRIEF SUMMARY OF THE INVENTION

[0014] Disclosed are compounds, compositions, and methods involving nanobodies that bind Henipavirus. In some forms, the nanobodies can neutralize Henipavirus. The disclosed nanobodies are generally useful for binding to and affecting Henipavirus, such as Hendra virus and Nipah virus. The disclosed nanobodies are useful for treating Henipavirus infections.

[0015] In some forms, the nanobody includes the complementarity-determining regions of nanobody ID nanobody ID NiVsF_Nbl, nanobody ID NiVsF_Nb2, nanobody ID NiVsF_Nb3, nanobody ID NiVsF_Nb4. nanobody ID NiVsF_Nb5, nanobody ID NiVsF_Nb6, nanobody ID NiVsF_Nb7, nanobody ID NiVsF_Nb8. nanobody ID NiVsF_Nb9, nanobody ID NiVsF_Nbl0, nanobody ID HeVsF_Nbl, nanobody ID HeVsF_Nb2, nanobody ID HeVsF_Nb3, nanobody ID HeVsF_Nb4, nanobody ID HeVsF_Nb5, or nanobody ID HeVsF_Nb6.

[0016] In some forms, the nanobody includes the complementarity-determining regions and one or more of the framework regions of nanobody ID nanobody ID NiVsF_Nbl, nanobody ID NiVsF_Nb2, nanobody ID NiVsF_Nb3, nanobody ID NiVsF_Nb4. nanobody ID NiVsF_Nb5, nanobody ID NiVsF_Nb6, nanobody ID NiVsF_Nb7, nanobody ID NiVsF_Nb8. nanobody ID NiVsF_Nb9, nanobody ID NiVsF_NblO, nanobody ID HeVsF_Nbl, nanobody ID HeVsF_Nb2, nanobody ID HeVsF_Nb3, nanobody ID HeVsF_Nb4, nanobody ID HeVsF_Nb5, or nanobody ID HeVsF_Nb6.

[0017] In some forms, the nanobody includes nanobody ID nanobody ID NiVsF_Nbl, nanobody ID NiVsF_Nb2, nanobody ID NiVsF_Nb3, nanobody ID NiVsF_Nb4. nanobody ID NiVsF_Nb5, nanobody ID NiVsF_Nb6, nanobody ID NiVsF_Nb7, nanobody ID NiVsF_Nb8. nanobody ID NiVsF_Nb9, nanobody ID NiVsF_NblO, nanobody ID HeVsF_Nbl, nanobody ID HeVsF_Nb2, nanobody ID HeVsF_Nb3, nanobody ID HeVsF_Nb4, nanobody ID HeVsF_Nb5, or nanobody ID HeVsF_Nb6.

[0018] In some forms, the nanobody includes the amino acid sequence of nanobody ID nanobody ID NiVsF_Nbl, nanobody ID NiVsF_Nb2, nanobody ID NiVsF_Nb3, nanobody ID NiVsF_Nb4. nanobody ID NiVsF_Nb5, nanobody ID NiVsF_Nb6, nanobody ID NiVsF_Nb7, nanobody ID NiVsF_Nb8. nanobody ID NiVsF_Nb9, nanobody ID NiVsF_NblO, nanobody ID HeVsF_Nbl, nanobody ID HeVsF_Nb2, nanobody ID HeVsF_Nb3, nanobody ID HeVsF_Nb4, nanobody ID HeVsF_Nb5, or nanobody ID HeVsF_Nb6.

[0019] In some forms, the nanobody has an amino acid sequence that is 70%, 75%, 80%, 85%, 90%, 95%, or 99% sequence identity with the amino acid sequence of nanobody ID nanobody ID NiVsF_Nbl, nanobody ID NiVsF_Nb2, nanobody ID NiVsF_Nb3, nanobody ID NiVsF_Nb4. nanobody ID NiVsF_Nb5, nanobody ID NiVsF_Nb6, nanobody ID NiVsF_Nb7, nanobody ID NiVsF_Nb8. nanobody ID NiVsF_Nb9, nanobody ID NiVsF_NblO, nanobody ID HeVsF_Nbl, nanobody ID HeVsF_Nb2, nanobody ID HeVsF_Nb3, nanobody ID HeVsF_Nb4, nanobody ID HeVsF_Nb5, or nanobody ID HeVsF_Nb6.

[0020] In some forms, the nanobody is humanized.

[0021] In some forms, the nanobody neutralizes Hendra virus. In some forms, the nanobody is nanobody ID HeVsF_Nb2, nanobody ID HeVsF_Nb3, nanobody ID HeVsF_Nb4, nanobody ID HeVsF_Nb5, and nanobody ID HeVsF_Nb6.

[0022] In some forms, the nanobody neutralizes Nipah virus. In some forms, the nanobody is nanobody ID nanobody ID NiVsF_Nbl, nanobody ID NiVsF_Nb2, nanobody ID NiVsF_Nb3, nanobody ID NiVsF_Nb4. nanobody ID NiVsF_Nb5, nanobody ID NiVsF_Nb6, nanobody ID NiVsF_Nb7, nanobody ID NiVsF_Nb8. nanobody ID NiVsF_Nb9, and nanobody ID NiVsF_NblO.

[0023] Also disclosed are composition that include one or more of the disclosed nanobodies. In some forms, the composition includes a pharmaceutically acceptable carrier.

[0024] Also disclosed are methods of using the disclosed nanobodies and nanobody compositions. In some forms, the methods involve treating a Henipavirus infection by administering to a subject infected with Henipavirus an effective amount of one or more of the disclosed nanobodies, one or more of the nanobody compositions, or combinations thereof.

[0025] In some forms, the methods involve treating a Hendra virus infection by administering to a subject infected with Hendra virus an effective amount of one or more of the disclosed nanobodies, one or more of the nanobody compositions, or combinations thereof.

[0026] In some forms, the methods involve treating a Nipah virus infection by administering to a subject infected with Nipah virus an effective amount of one or more of the disclosed nanobodies, one or more of the nanobody compositions, or combinations thereof.

[0027] Additional advantages of the disclosed method and compositions will be set forth in part in the description which follows, and in part will be understood from the description, or can be learned by practice of the disclosed method and compositions. The advantages of the disclosed method and compositions will be realized and attained by means of the elements and combinations particularly pointed out in the appended claims. It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the invention as claimed.

[0028] BRIEF DESCRIPTION OF THE DRAWINGS

[0029] The accompanying drawings illustrate several embodiments of the disclosed method and compositions and together with the description, serve to explain the principles of the disclosed method and compositions.

[0030] Figure 1 is a diagram showing features of Henipavirus, their range, and their public health significance.

[0031] Figure 2 is a diagram of the vectors and transmission pathways of Henipavirus.

[0032] Figure 3 is a diagram showing the pathogenic pathways and effects of Nipah virus.

[0033] Figure 4 is a diagram showing structural features of Henipavirus and the Henipavirus lifecycle in cells.

[0034] Figure 5 is diagrams showing comparative structures of conventional antibodies and nanobodies (top) and features of nanomouse systems for producing nanobodies (bottom).

[0035] Figure 6 is a table comparing characteristics of conventional antibodies and nanobodies. Figure 7 is a diagram illustrating the process for generating nanobodies, with the production of nanobodies to the F protein of HeV and NiV show.

[0036] Figure 8 shows the results of examples of generated nanobodies binding to the F protein.

[0037] Figure 9 is a table binding constants (KD (M), kon(l / M), and kdis(l / s))) measured for example of generated nanobodies.

[0038] Figure 10 is a diagram of the production of VS V pseudotyped viruses bearing Henipavirus F and G proteins.

[0039] Figures 11A-11C show example results of VSV pseudovirus infections.

[0040] Figures 12A-12C show example results of functional assays of neutralization of VSV pseudovirus infections, CedV chimeric virus infections, and authentic virus infections by NiV and HeV nanobodies. Figures 12A and 12B are neutralization curves. Figure 12 Cis a table showing the IC50 of NiV and HeV against VSVpp, CedV chimeric virus, and authentic virus.

[0041] Figures 13A-13C show results of neutralization of HeV and NiV VSV pseudoviruses by examples of HeV nanobodies. The HeV nanobodies cross-neutralize HeV and NiV VSV pseudoviruses.

[0042] Figures 14A- 14C show results of neutralization of HeV and NiV VSV pseudoviruses by examples of NiV nanobodies. The NiV nanobodies cross-neutralize HeV and NiV VSV pseudoviruses.

[0043] Figures 15A and 15B show results of neutralization of HeV and NiV chimeric CedV by examples of HeV nanobodies. Most of the nanobodies cross-neutralize chimeric CedV.

[0044] Figures 16A and 16B show results of neutralization of HeV and NiV VSV chimeric CedV by examples of NiV nanobodies. Most of the nanobodies cross-neutralize chimeric CedV.

[0045] Figures 17A-17C show results of neutralization of cell-cell fusion of HeV and NiV by examples of HeV nanobodies.

[0046] Figures 18A-18C show results of neutralization of cell-cell fusion of HeV and NiV by examples of NiV nanobodies. The NiV nanobodies cross-neutralize HeV and NiV VSV pseudoviruses.

[0047] Figure 19 is a diagram of the structure of monomer, monomer bivalent, dimer bivalent, and trimer bivalent nanobodies.

[0048] Figure 20 is a gel of the nanobody structures of Fig. 19.

[0049] Figures 21 A and 21B are diagrams of the ribbon structure of and example nanobody (NblO) binding to an NiV F trimer. Fig. 21 A is a top view. Fig. 21B is a side view. DETAILED DESCRIPTION OF THE INVENTION

[0050] The disclosed method and compositions can be understood more readily by reference to the following detailed description of particular embodiments and the Example included therein and to the Figures and their previous and following description.

[0051] It is to be understood that the disclosed method and compositions are not limited to specific synthetic methods, specific analytical techniques, or to particular reagents unless otherwise specified, and, as such, can vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting.

[0052] It is to be understood that the disclosed compounds, compositions, and methods are not limited to specific synthetic methods, specific analytical techniques, or to particular reagents unless otherwise specified, and, as such, may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular forms and embodiments only and is not intended to be limiting.

[0053] A. Nanobodies

[0054] The disclosed nanobodies are particularly useful in (and intended for) pharmaceutical uses, and in some forms, they can have a high degree of sequence homology in their framework regions with the framework sequences of human VH domains. In some forms, the nanobodies can have an overall degree of sequence identity (determined as further described herein, and taking into account only the framework regions and not the CDR's) with at least one human germline sequence of at least 80%, preferably at least 85%, such as 90% or more. In some forms, the nanobody can have an overall degree of sequence identity (determined as further described herein, and taking into account only the framework regions and not the CDR’s) of at least 80%, preferably at least 85%, such as 90% or more with at least one of the following human germline sequences: DP-47, DP-51 and / or DP-29. The DP47 germline sequence for the heavy chain variable domain, which confers high thermal stability and protein A binding properties.

[0055] However, according to a preferred aspect of the invention, the VH domain of the invention is an (heavy-chain) immunoglobulin single variable domain or “ISVD”, meaning a heavy-chain variable domain that can form a functional antigen binding site without interaction with a VL domain. For example, the VH domain of the invention can be a The nanobody be, or be derived from, a VHH, a humanized VHH and / or a camelized VHs such as camelized human VH's.

[0056] The term “nanobody” generally refers to a VHH, a humanized VHH or a camelized VH (such as a camelized human VH) or generally a sequence optimized VHH (such as e.g. optimized for chemical stability and / or solubility, maximum overlap with known human framework regions and maximum expression).

[0057] The term “ISVD” (or “ISV”) as used herein in its broadest sense also includes “TSVD- based biologicals” and, when the ISVD is a nanobody, “nanobody-based biologicals”. An “ISVD-based biological” is defined herein as a protein, polypeptide or other biological drug that comprises or essentially consist of at least one (such as one, two or three) ISVD's. Similarly, a “nanobody-based biological” is defined as a protein, polypeptide or other biological drug that comprises or consists essentially of at least one (such as one, two or three) nanobodies. As with the term “ISVD”, whenever the term “ISVD-based biological” is used, it should be understood that such an ISVD-based biological is preferably a nanobody -based biological. As disclosed herein, both an “ISVD-based biological” and a “nanobody-based biological” may for example be a monovalent, bivalent (or multivalent), bispecific (or multispecific), and biparatopic (or multiparatopic) ISVD construct or nanobody construct, respectively. Also, any ISVD-based or nanobody-based biological may for example, in addition to the one or more (such as one, two or three) ISVD's or nanobodies, optionally further comprise one or more (such as one or two) other further therapeutic moieties and / or one or more (such as one or two) other moieties that influence the pharmacokinetic or pharmacodynamic properties of the ISVD-based or nanobody- based biological (such as its half-life).

[0058] Suitable examples of such further therapeutic or other moieties will be clear to the skilled person, and for example generally can include any therapeutically active protein, polypeptide or other binding domain or binding unit, as well as for example modifications such as those described on pages 149 to 152 of WO 09 / 138159. An ISVD-based biological or nanobody-based biological is preferably a therapeutic or intended for use as a therapeutic (which includes prophylaxis and diagnosis) and for this purpose preferably contains at least one ISVD against a therapeutically relevant target.

[0059] As used herein, the amino acid residues of a nanobody are numbered according to the general numbering for VH domains given by Kabat et al. (“Sequence of proteins of immunological interest”, US Public Health Services, NIH Bethesda, Md., Publication No. 91), as applied to VHH domains from Camelids in the article of Riechmann and Muyldermans, J. Immunol. Methods 2000 Jun. 23; 240 (1-2): 185-195; or referred to herein. According to this numbering, FR1 of a nanobody comprises the amino acid residues at positions 1-30, CDR1 of a nanobody comprises the amino acid residues at positions 31-35, FR2 of a nanobody comprises the amino acids at positions 36-49, CDR2 of a nanobody comprises the amino acid residues at positions 50-65, FR3 of a nanobody comprises the amino acid residues at positions 66-94, CDR3 of a nanobody comprises the amino acid residues at positions 95-102, and FR4 of a nanobody comprises the amino acid residues at positions 103-113. In this respect, it should be noted that — as is well known in the art for VH domains and for VHH domains — the total number of amino acid residues in each of the CDR's may vary and may not correspond to the total number of amino acid residues indicated by the Kabat numbering (that is, one or more positions according to the Kabat numbering may not be occupied in the actual sequence, or the actual sequence may contain more amino acid residues than the number allowed for by the Kabat numbering). This means that, generally, the numbering according to Kabat may or may not correspond to the actual numbering of the amino acid residues in the actual sequence. Generally, however, it can be said that, according to the numbering of Kabat and irrespective of the number of amino acid residues in the CDR’s, position 1 according to the Kabat numbering corresponds to the start of FR1 and vice versa, position 36 according to the Kabat numbering corresponds to the start of FR2 and vice versa, position 66 according to the Kabat numbering corresponds to the start of FR3 and vice versa, and position 103 according to the Kabat numbering corresponds to the start of FR4 and vice versa.

[0060] Alternative methods for numbering the amino acid residues of VH domains, which methods can also be applied in an analogous manner to VHH domains from Camelids and to nanobodies, are the method described by Chothia et al. (Nature 342, 877-883 (1989)), the so- called “AbM definition” and the so-called “contact definition”.

[0061] In some forms, the C-terminal extension present in the nanobody does not contain a (free) cysteine residue (unless said cysteine residue is used or intended for further functionalization, for example for pegylation). In some forms, the preferences that are indicated on pages 35 to 41 of WO 12 / 175741 for the C-terminal extensions used according to WO 12 / 175741 can be used in the C-terminal extensions used in the nanobodies. In some forms, when the nanobody contains a C-terminal extension (X)n, n=l, 2 or 3, and each X is either Ala of Gly. In some forms, each X is Ala, and n=l or 2, and preferably 1.

[0062] Nanobodies can be easily assembled into multimers, leading to increased avidity (Fig. 19). Multimeric nanobodies exhibit higher binding affinity and stronger neutralization potency. Multimeric nanobodies exhibit higher binding affinity and stronger neutralization potency.

[0063] B. Administration

[0064] The term “hit” refers to a test compound that shows desired properties in an assay. The term “test compound” refers to a chemical to be tested by one or more screening method(s) as a putative modulator. A test compound can be any chemical, such as an inorganic chemical, an organic chemical, a protein, a peptide, a carbohydrate, a lipid, or a combination thereof. Usually, various predetermined concentrations of test compounds are used for screening, such as 0.01 micromolar, 1 micromolar and 10 micromolar. Test compound controls can include the measurement of a signal in the absence of the test compound or comparison to a compound known to modulate the target.

[0065] The terms “high,” “higher,” “increases,” “elevates,” or “elevation” refer to increases above basal levels, e.g., as compared to a control. The terms “low,” “lower,” “reduces,” or “reduction” refer to decreases below basal levels, e.g., as compared to a control.

[0066] The term “modulate” as used herein refers to the ability of a compound to change an activity in some measurable way as compared to an appropriate control. As a result of the presence of compounds in the assays, activities can increase or decrease as compared to controls in the absence of these compounds. Preferably, an increase in activity is at least 25%, more preferably at least 50%, most preferably at least 100% compared to the level of activity in the absence of the compound. Similarly, a decrease in activity is preferably at least 25%, more preferably at least 50%, most preferably at least 100% compared to the level of activity in the absence of the compound. A compound that increases a known activity is an “agonist.” One that decreases, or prevents, a known activity is an “antagonist.”

[0067] The term “inhibit” means to reduce or decrease in activity or expression. This can be a complete inhibition of activity or expression, or a partial inhibition. Inhibition can be compared to a control or to a standard level. Inhibition can be 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15,

[0068] 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41,

[0069] 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64,65, 66, 67,

[0070] 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93,

[0071] 94, 95, 96, 97, 98, 99, or 100%.

[0072] The term “monitoring” as used herein refers to any method in the art by which an activity can be measured.

[0073] The term “providing” as used herein refers to any means of adding a compound or molecule to something known in the art. Examples of providing can include the use of pipettes, pipettemen, syringes, needles, tubing, guns, etc. This can be manual or automated. It can include transfection by any mean or any other means of providing nucleic acids to dishes, cells, tissue, cell-free systems and can be in vitro or in vivo.

[0074] The term “preventing” as used herein refers to administering a compound prior to the onset of clinical symptoms of a disease or conditions so as to prevent a physical manifestation of aberrations associated with the disease or condition.

[0075] The term “in need of treatment” as used herein refers to a judgment made by a caregiver (e.g. physician, nurse, nurse practitioner, or individual in the case of humans; veterinarian in the case of animals, including non-human mammals) that a subject requires or will benefit from treatment. This judgment is made based on a variety of factors that are in the realm of a care giver’s expertise, but that include the knowledge that the subject is ill, or will be ill, as the result of a condition that is treatable by the disclosed compounds.

[0076] As used herein, “subject” includes, but is not limited to, animals, plants, bacteria, viruses, parasites and any other organism or entity. The subject can be a vertebrate, more specifically a mammal (e.g., a human, horse, pig, rabbit, dog, sheep, goat, non-human primate, cow, cat, guinea pig or rodent), a fish, a bird or a reptile or an amphibian. The subject can be an invertebrate, more specifically an arthropod (e.g., insects and crustaceans). The term does not denote a particular age or sex. Thus, adult and newborn subjects, as well as fetuses, whether male or female, are intended to be covered. A patient refers to a subject afflicted with a disease or disorder. The term “patient” includes human and veterinary subjects.

[0077] By “treatment” and “treating” is meant the medical management of a subject with the intent to cure, ameliorate, stabilize, or prevent a disease, pathological condition, or disorder. This term includes active treatment, that is, treatment directed specifically toward the improvement of a disease, pathological condition, or disorder, and also includes causal treatment, that is, treatment directed toward removal of the cause of the associated disease, pathological condition, or disorder. Tn addition, this term includes palliative treatment, that is, treatment designed for the relief of symptoms rather than the curing of the disease, pathological condition, or disorder; preventative treatment, that is, treatment directed to minimizing or partially or completely inhibiting the development of the associated disease, pathological condition, or disorder; and supportive treatment, that is, treatment employed to supplement another specific therapy directed toward the improvement of the associated disease, pathological condition, or disorder. It is understood that treatment, while intended to cure, ameliorate, stabilize, or prevent a disease, pathological condition, or disorder, need not actually result in the cure, amelioration, stabilization or prevention. The effects of treatment can be measured or assessed as described herein and as known in the art as is suitable for the disease, pathological condition, or disorder involved. Such measurements and assessments can be made in qualitative and / or quantitative terms. Thus, for example, characteristics or features of a disease, pathological condition, or disorder and / or symptoms of a disease, pathological condition, or disorder can be reduced to any effect or to any amount.

[0078] A cell can be in vitro. Alternatively, a cell can be in vivo and can be found in a subject. A “cell” can be a cell from any organism including, but not limited to, a bacterium.

[0079] In one aspect, the compounds described herein can be administered to a subject comprising a human or an animal including, but not limited to, a mouse, dog, cat, horse, bovine or ovine and the like, that is in need of alleviation or amelioration from a recognized medical condition. By the term “effective amount” of a compound as provided herein is meant a nontoxic but sufficient amount of the compound to provide the desired result. As will be pointed out below, the exact amount required will vary from subject to subject, depending on the species, age, and general condition of the subject, the severity of the disease that is being treated, the particular compound used, its mode of administration, and the like. Thus, it is not possible to specify an exact “effective amount.” However, an appropriate effective amount can be determined by one of ordinary skill in the art using only routine experimentation.

[0080] The dosages or amounts of the compounds described herein are large enough to produce the desired effect in the method by which delivery occurs. The dosage should not be so large as to cause adverse side effects, such as unwanted cross-reactions, anaphylactic reactions, and the like. Generally, the dosage will vary with the age, condition, sex and extent of the disease in the subject and can be determined by one of skill in the art. The dosage can be adjusted by the individual physician based on the clinical condition of the subject involved. The dose, schedule of doses and route of administration can be varied.

[0081] The efficacy of administration of a particular dose of the compounds or compositions according to the methods described herein can be determined by evaluating the particular aspects of the medical history, signs, symptoms, and objective laboratory tests that are known to be useful in evaluating the status of a subject in need for the treatment of Henipavirus infection or other diseases and / or conditions. These signs, symptoms, and objective laboratory tests will vary, depending upon the particular disease or condition being treated or prevented, as will be known to any clinician who treats such patients or a researcher conducting experimentation in this field. For example, if, based on a comparison with an appropriate control group and / or knowledge of the normal progression of the disease in the general population or the particular individual: (1) a subject’s physical condition is shown to be improved (e.g., a tumor has partially or fully regressed), (2) the progression of the disease or condition is shown to be stabilized, or slowed, or reversed, or (3) the need for other medications for treating the disease or condition is lessened or obviated, then a particular treatment regimen will be considered efficacious.

[0082] By “pharmaceutically acceptable” is meant a material that is not biologically or otherwise undesirable, i.e., the material can be administered to a subject along with the selected compound without causing any undesirable biological effects or interacting in a deleterious manner with any of the other components of the pharmaceutical composition in which it is contained.

[0083] Any of the compounds having the formula I can be used therapeutically in combination with a pharmaceutically acceptable carrier. The compounds described herein can be conveniently formulated into pharmaceutical compositions composed of one or more of the compounds in association with a pharmaceutically acceptable carrier. See, e.g., Remington's Pharmaceutical Sciences, latest edition, by E.W. Martin Mack Pub. Co., Easton, PA, which discloses typical carriers and conventional methods of preparing pharmaceutical compositions that can be used in conjunction with the preparation of formulations of the compounds described herein. These most typically would be standard carriers for administration of compositions to humans. In one aspect, humans and non-humans, including solutions such as sterile water, saline, and buffered solutions at physiological pH. Other compounds will be administered according to standard procedures used by those skilled in the art.

[0084] The pharmaceutical compositions described herein can include, but are not limited to, carriers, thickeners, diluents, buffers, preservatives, surface active agents and the like in addition to the molecule of choice. Pharmaceutical compositions can also include one or more active ingredients such as antimicrobial agents, antiinflammatory agents, anesthetics, and the like.

[0085] The compounds and pharmaceutical compositions described herein can be administered to the subject in a number of ways depending on whether local or systemic treatment is desired, and on the area to be treated. Moreover, a pharmaceutical composition can be administered to a subject vaginally, rectally, intranasally, orally, by inhalation, or parenterally, for example, by intradermal, subcutaneous, intramuscular, intraperitoneal, intrarectal, intraarterial, intralymphatic, intravenous, intrathecal and intratracheal routes. Parenteral administration, if used, is generally characterized by injection and includes intravenous (IV), subcutaneous (SC), intramuscular (IM), epidural and intra- articular injection, as well as surgical insertion of depots in the organ or tissue of interest (Bittner, et al., BioDrugs., 32:425-440 (2018); Lee et al., J. Pharm. Investig., 49: 459-476 (2019); Chaudhary et al., Crit. Rev. Ther. Drug Carrier Syst., 36: 137-181 (2019)). Injectables can be prepared in conventional forms, either as liquid solutions or suspensions, solid forms suitable for solution or suspension in liquid prior to injection, or as emulsions (Park et al., J Control Release, 342:53-65 (2022); Nkanga, et al., Advanced Drug Delivery Reviews, 167:19-46, (2020); Sheikh, et al., Asian Journal of Pharmaceutics, 10(4):S465-S471 (2016); Rhee et al., Pharmaceutical Technology Drug Delivery, p.S6 (2010)). An exemplary approach for parenteral administration involves use of a slow release or sustained release system such that a constant dosage is maintained. See for example, U.S. Patent No. 9,700,630, WO 2006 / 125620, and KR 101898816.

[0086] Preparations for parenteral administration include sterile aqueous or non-aqueous solutions, suspensions, and emulsions which can also contain buffers, diluents and other suitable additives. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Aqueous carriers include water, alcoholic / aqueous solutions, emulsions or suspensions, including saline and buffered media. Parenteral vehicles include sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride, lactated Ringer's, or fixed oils. Intravenous vehicles include fluid and nutrient replenishers, electrolyte replenishers (such as those based on Ringer's dextrose), and the like. Preservatives and other additives can also be present such as, for example, antimicrobials, anti-oxidants, chelating agents, and inert gases and the like.

[0087] Formulations for topical administration can include ointments, lotions, creams, gels, drops, suppositories, sprays, liquids and powders. Conventional pharmaceutical carriers, aqueous, powder or oily bases, thickeners and the like can be necessary or desirable.

[0088] Compositions for oral administration can include powders or granules, suspensions or solutions in water or non-aqueous media, capsules, sachets, or tablets. Thickeners, flavorings, diluents, emulsifiers, dispersing aids or binders can be desirable.

[0089] Sequences

[0090] SEQ ID NO: 1 - DNA sequences of NiVsFJNbl

[0091] CAGGTGCAATTGGTAGAGTCTGGCGGTGGACTCGTTCAAGCCGGAGGCTCACTGCG GTTGTCTTGTGCCGCATCTGGTCGAACCTTCTCAAGTTATGCTATGGGGTGGTTCCG GCAGGCTCCTGGTAAAGAGCGGGAATTTGTCGCAGCAATTGGTTGGTCTGGTGCCA ATATTTACTATGCTGATTCAGTAAAAGGTCGATTCACTATATCACGGGATAATGCAA AAAATACTGTCTATCTCCAAATGAACTCTCTGAAGCCTGAAGATACTGCTGTGTATT ACTGTGTCGCAGATTCTTTTCTTCCATTACTACGGCTACGAGGGGGTGACTACTGGG GCCAAGGCACCACTCTCACAGTCTCCTCG

[0092] SEQ ID NO: 2 - Amino acid sequences of NiVsF_Nbl

[0093] QVQLVESGGGLVQAGGSLRLSCAASGRTFSSYAMGWFRQAPGKEREFVAAIGWSGANI YYADSVKGRFTISRDNAKNTVYLQMNSLKPEDTAVYYCVADSFLPLLRLRGGDYWGQ GTTLTVSS

[0094] SEQ ID NO: 3 - DNA sequences of NiVsF_Nb2

[0095] CAGGTTCAACTCGTTGAGTCAGGCGGTGGACTCGTTCAAGCCGGAGGCTCACTGCG GTTGTCTTGTGCCGCATCTGGTCGAACCTTCTCAAGTTATGCTATGGGGTGGTTCCG GCAGGCTCCAGGTAAAGAGCGGGAATTTGTCGCAGCAATTAGTTGGTCTGGTGGTA GTGCTTACTATACTGATTCAGTAAAAGGTCGATTCACTATATCACGGGATAATGAAA AAAATACTGTCTATCTCCAAATGAACTCTCTGACGCCTGAAGATACTGCTGTGTATT CCTGTGCCGGAGGAGAGGCTCACTCCTACTATAGGCCCAACTCTGCTATGGACTACT GGGGTCAAGGAACCTCAGTCACCGTCTCCTCG

[0096] SEQ ID NO: 4 - Amino acid sequences of NiVsF_Nb2

[0097] QVQLVESGGGLVQAGGSLRLSCAASGRTFSSYAMGWFRQAPGKEREFVAAISWSGGSA YYTDSVKGRFTISRDNEKNTVYLQMNSLTPEDTAVYSCAGGEAHSYYRPNSAMDYWG QGTSVTVSS SEQ ID NO: 5 - DNA sequences of NiVsF_Nb3

[0098] GAAGTCCAACTTGTAGAGTCAGGCGGTGGACTCGTTCAAGCCGGAGGCTCACTGCG

[0099] GTTGTCTTGTGCCGCATCTGGTCGAACCTTCTCAAGTTATGTTATGGGGTGGTTCCGG

[0100] CAGGCTCCNGGTAAAGAGCGGGAATTTGTCGCAGCAATTGGTTGGTCTGGTGGTAA

[0101] TACTTACTCTTCTGCTTCAGTAAAAGGTCGATTCACTATATCACGGGATAATTCAAA

[0102] AAATACTGTCTATCTCCAAATGAACTCTCTGAAGCCTGAAGATACTGCTGTGTATTA

[0103] CTGTGCCGCAGGAAGGGTGTCCTACTATAGGTACCACCCCTTTGACTACTGGGGCCA

[0104] AGGCACCACTCTCACAGTCTCCTCG

[0105] SEQ ID NO: 6 - Amino acid sequences of NiVsF_Nb3

[0106] EVQLVESGGGLVQAGGSLRLSCAASGRTFSSYVMGWFRQAPGKEREFVAAIGWSGGNT YSSASVKGRFTISRDNSKNTVYLQMNSLKPEDTAVYYCAAGRVSYYRYHPFDYWGQG TTLTVSS

[0107] SEQ ID NO: 7 - DNA sequences of NiVsF_Nb4

[0108] CAAGTTCAGCTTGTAGAGTCTGGCGGTGGACTCGTTCAAGCCGGAGGCTCACTGCG

[0109] GTTGTCTTGTGCCGCATCTGGTCGAACCTTCTCAAGTTATGCTATGGGGTGGTTCCG

[0110] GCAGGCTCCAGGTAAAGAGCGGGAATTTGTCGCAGGAATAAGTTGGTCTGGTGTTA

[0111] ATACTTACTATGCTGACTCAGTGCAAGGTCGATTCACTATATCACGGGATAATGCAA

[0112] AAAATACTGTCTATCTCCAAATGAACTCTCTGAAGCCTGAAGATACTGCTGTGTATT

[0113] ACTGTGCCGCAGATCGGGCCTCGGCCTACTATGGTCTACATTACTATGCTATGGCCT

[0114] ACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCG

[0115] SEQ ID NO: 8 - Amino acid sequences of NiVsF_Nb4

[0116] QVQLVESGGGLVQAGGSLRLSCAASGRTFSSYAMGWFRQAPGKEREFVAGISWSGVNT

[0117] YYADSVQGRFTISRDNAKNTVYLQMNSLKPEDTAVYYCAADRASAYYGLHYYAMAY WGQGTSVTVSS

[0118] SEQ ID NO: 9 - DNA sequences of NiVsF_Nb5

[0119] CAGGTCCAACTGGTAGAGTCAGGCGGTGGACTCGTTCAAGCCGGAGGCTCACTGCG

[0120] GTTGTCTTGTGCCGCATCTGGTCGAACCTTCTCAAGTTATGCAATGGGGTGGTTCCG

[0121] GCAGGCTCCTGGTAAAGAGCGGGAATTTGTCGCAGCAATTAGTTGGTCTGGTGGTA

[0122] GTTCTTACTACGCTGATTCAGTAAAAGGTCGATTCACTATATCACGGGATAATGCAA

[0123] AAAATACTGGCTATCTCCAAATGAGCTCTCTGAAGCCTGAAGATACTGCTGTGTATT ACTGTGCCGCAGGCGCGCCGCACACTTACTACAGTAGTCGGCACCATGCTATGGACT ACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCG SEQ ID NO: 10 - Amino acid sequences of NiVsF_Nb5

[0124] QVQLVESGGGLVQAGGSLRLSCAASGRTFSSYAMGWFRQAPGKEREFVAAISWSGGSS

[0125] YYADSVKGRFTISRDNAKNTGYLQMSSLKPEDTAVYYCAAGAPHTYYSSRHHAMDYW GQGTSVTVSS

[0126] SEQ ID NO: 11 - DNA sequences of NiVsF_Nb6

[0127] CAGGTGCAATTGGTAGAGTCTGGCGGTGGACTCGTTCAAGCCGGAGACTCACTGCG

[0128] GTTGTCTTGTGCCGCATCTGGTCGATCCTTCTCAAGTTTTGTTATGGGGTGGTTCCGG

[0129] CAGGCTCCAGGTACAGAGCGGGAATTTGTCGCAGCAATTAGTTGGTCTGGTGGTAG

[0130] TACTTACTCTTCTGATTCAGTAAAAGGTCGATTCACTATATCACGGGATAATTCAAA

[0131] AAATACTGTCTATCTCCAAATGAAGTCTCTGAAGCCTGAAGATACTGCTGTGTATTA

[0132] CTGTGCCGCAGGAAGGGTGTCCTACGATAGGTACGACGCCTTTGACTACTGGGGCC

[0133] AAGGCACCACTCTCACAGTCTCCTCG

[0134] SEQ ID NO: 12 - Amino acid sequences of NiVsF_Nb6

[0135] QVQLVESGGGLVQAGDSLRLSCAASGRSFSSFVMGWFRQAPGTEREFVAAISWSGGST

[0136] YSSDSVKGRFTISRDNSKNTVYLQMKSLKPEDTAVYYCAAGRVSYDRYDAFDYWGQG TTLTVSS

[0137] SEQ ID NO: 13 - DNA sequences of NiVsF_Nb7

[0138] CAGGTGCAATTGGTAGAGTCTGGCGGTGGACTCGTTCAAGCCGGAGGCTCACTGCG

[0139] GTTGTCTTGTGCCGCATCTGGTCGAACCTTCTCAAGTTATGCTATGGGGTGGTTCCG

[0140] GCAGGCTCCAAATAAAGAGCGGGAATTTGTCGCAGCAATTAGTTGGTCTGGTGGTA

[0141] GTTCTTACTATGTTGATTCAGTAAAAGGTCGATTCACTATATCACGGGATAATGCAA

[0142] TAAATACTGTCTATCTCCAAATGAACTCTCTGAAGCCTGAAGATACTGCTGTGTATT

[0143] TCTGTGCCGGAGGAGAGGCTCACTCCTACTATAGGCCCTACTATGCTATGGACTACT

[0144] GGGGTCAAGGAACCTCAGTCACCGTCTCCTCG

[0145] SEQ ID NO: 14 - Amino acid sequences of NiVsF_Nb7

[0146] QVQLVESGGGLVQAGGSLRLSCAASGRTFSSYAMGWFRQAPNKEREFVAAISWSGGSS

[0147] YYVDSVKGRFTISRDNAINTVYLQMNSLKPEDTAVYFCAGGEAHSYYRPYYAMDYWG QGTSVTVSS

[0148] SEQ ID NO: 15 - DNA sequences of NiVsF_Nb8

[0149] CAAGTCCAGCTCGTCGAGTCAGGCGGTGGACTCGTTCAAGCCGGAGGCTCACTGCG

[0150] GTTGTCTTGTGCCGCATCTGGTCGAGCCTTCTCAAGTTATGCTGTGGGGTGGTTCCG

[0151] GCAGGCTCCAGGTAAAGAGCGGGAATTTGTCGCAGCAATTAGTTGGTCTGGTGGTA

[0152] GTTCTTACCAGGCTGATTCAGTAAAAGGTCGATTCACTGTATCACGGGATATTGCAA

[0153] AAAATACTGTCTATCTCCAAATGAACTCTCTGAAGCCTGAAGATACTGCTGTGTATT ACTGTGCCGCAGATCGGGCCTCGGCCTACTATACTCTACATTACTATACTATGGACT ACTGGGGTCAAGGAACCACGGTCACCGTCTCCTCG

[0154] SEQ ID NO: 16 - Amino acid sequences of NiVsF_Nb8

[0155] QVQLVESGGGLVQAGGSLRLSCAASGRAFSSYAVGWFRQAPGKEREFVAAISWSGGSS

[0156] YQADSVKGRFTVSRDIAKNTVYLQMNSLKPEDTAVYYCAADRASAYYTLHYYTMDY WGQGTTVTVSS

[0157] SEQ ID NO: 17 - DNA sequences of NiVsF_Nb9

[0158] CAAGTTCAACTCGTAGAATCTGGCGGTGGACTCGTTCAGGCCGGAGGCTCACTGCG

[0159] GTTGTCTTGTGCCGCATCTGGTCGAACCTTCTCAAGTTATGCTGTGGGGTGGTACCG

[0160] GCAGGCTCCAGGTAAAGAGCGGGAATTTGTCGCAGCAATTAGTTGGTCTGGTGTTA

[0161] ATACTTACTATGCTGATTCAGTAAAAGGTCGATTCACTATATCGCGGGATAATGTGA

[0162] AGAATGCTGTCTATCTCCAAATGAACTCTCTGAAGCCTGAAGATACTGCTGTGTATT

[0163] ACTGTGGCGCAGATCGGGCCTCGGCCTACTATGGTCTATATTACTCTGTTCCGGAGT

[0164] ACTGGGGTCAGGGAACCTCAGTCACCGTCTCCTCG

[0165] SEQ ID NO: 18 - Amino acid sequences of NiVsF_Nb9

[0166] QVQLVESGGGLVQAGGSLRLSCAASGRTFSSYAVGWYRQAPGKEREFVAAISWSGVNT

[0167] YYADSVKGRFTISRDNVKNAVYLQMNSLKPEDTAVYYCGADRASAYYGLYYSVPEYW GQGTSVTVSS

[0168] SEQ ID NO: 19 - DNA sequences of NiVsI Nb 10

[0169] CAAGTAAAACTTGAAGAATCAGGTGGTGGACTCGTTCAAGCCGGAGGCTCACTGCG

[0170] GTTGTCTTGTGCCGCATCTGGTCGAACCTTCTCAAGTTATGCTATGGGGTGGTTCCG

[0171] GCAGGCTCCAGATAAAGAGCGGGAATTTGTCGCAGCAATAAGTTGGTCTGGTGTTA

[0172] ATACTTACTATGCTGATTCAGTGCAAGGTCGATTCACTGTATCACGGGATAATGCAA

[0173] AAAATACTGTCTATCTCCAAATGAACTCTCTGAAGCCTGAAGATACTGCTGTGTATT

[0174] ACTGTGCCGGAGGAGAGGCTCACTCCTACTATAGGCCCTACTATGCTATGGACTACT

[0175] GGGGGCAAGGAACCTCAGTCACCGTCTCCTCG

[0176] SEQ ID NO: 20 - Amino acid sequences of NiVsF_NblO

[0177] QVKLEESGGGLVQAGGSLRLSCAASGRTFSSYAMGWFRQAPDKEREFVAAISWSGVNT

[0178] YYADSVQGRFTVSRDNAKNTVYLQMNSLKPEDTAVYYCAGGEAHSYYRPYYAMDYW GQGTSVTVSS

[0179] SEQ ID NO: 21 - DNA sequences of HeVsF_Nbl

[0180] CAAGTTCAACTTGTTGAATCAGGTGGTGGACTCGTTCAAGCCGGAGGCTCACTGCGG

[0181] TTGTCTTGTGCCGCATCTGGTCGAACCTTCTCAAGTTATGCTTTGGGGTGGTTCCGGC

[0182] AGGCTCCAGGTAAAGAGCGGGAATTTGTCGCAGCAATTAGTTGGTCTGGTGTTAAT

[0183] ACTTACTATGCTGATTCAGTAAAAGGTCGATTCACTATATCACGGGATGATGCAAGA AATACTGTCTATCTCCAAATGAACTCTCTGAAGCCTGAAGATACTGCTGTGTATTAC

[0184] TGTGCCGCAGGGGGGCCCCCGGCCTACTATAGGTACGAAACTATGGACTACTGGGG

[0185] TCAAGGAACCTCAGTCACCGTCTCCTCG

[0186] SEQ ID NO: 22 - Amino acid sequences of HeVsF_Nbl

[0187] QVQLVESGGGLVQAGGSLRLSCAASGRTFSSYALGWFRQAPGKEREFVAAISWSGVNT

[0188] YYADSVKGRFTISRDDARNTVYLQMNSLKPEDTAVYYCAAGGPPAYYRYETMDYWG QGTSVTVSS

[0189] SEQ ID NO: 23 - DNA sequences of HeVsF_Nb2

[0190] CAGGTCCAACTGGTAGAGTCAGGCGGTGGACTCGTTCAAGCCGGAGGCTCACTGCG

[0191] GTTGTCTTGTGCCGCATCTGGTCGAAACTTCTCAAGTTATGCTATGGGGTGGTTCCG

[0192] GCAGGCTCCAGGAAAAGAGCGGGAATTTGTCGCAGCAATTAGTTGGTCTGGTGTTA

[0193] ATACTTACTATGCTGATTCAGTAAAAGGTCGATTCACTATATCACGGGATAATGAAA

[0194] AAAATACTGCCCATCTCCAAATGAACTCTCTGAAGCCTGAAGACTCTGCTGTGTATT

[0195] ACTGTGCCGCAGGGGAGCCCCCGGCCTACTATAGATACGAAACTATGGACCACTGG

[0196] GGTCAAGGAACCTCAGTCACCGTCTCCTCG

[0197] SEQ ID NO: 24 - Amino acid sequences of HeVsF_Nb2

[0198] QVQLVESGGGLVQAGGSLRLSCAASGRNFSSYAMGWFRQAPGKEREFVAAISWSGVN

[0199] TYYADSVKGRFTISRDNEKNTAHLQMNSLKPEDSAVYYCAAGEPPAYYRYETMDHWG QGTSVTVSS

[0200] SEQ ID NO: 25 - DNA sequences of HeVsF_Nb3

[0201] CAAGTAAAACTTGAAGAATCAGGTGGTGGACTCGTTCAAGCCGGAGGCTCACTGCG

[0202] GTTGTCTTGTGCCGCATCTGGTCGAACCTTCTCAAGTTATGCTATGGGGTGGTTCCG

[0203] GCAGGCTCCAGGGAAAGAGCGGGAATTTGTCGCAGTAATTAGTTGGTCTGGTGGTA

[0204] GTGCTTACTATGCTGATTCAGTAAAAGGTCGATTCACTATATCACGGGATAATGCAG

[0205] AAAATACTGTCTATCTCCAAATGAACTCTCTGAAGCCTGAAGATACTGCTGTGTATT

[0206] ACTGTGCCGCAGGGGGGCCCCCGGCCTACTATAGGTACGATACTATGGACTACTGG

[0207] GGTCAAGGAACCACGGTCACCGTCTCCTCG

[0208] SEQ ID NO: 26 - Amino acid sequences of HeVsF_Nb3

[0209] QVKLEESGGGLVQAGGSLRLSCAASGRTFSSYAMGWFRQAPGKEREFVAVISWSGGSA

[0210] YYADSVKGRFTISRDNAENTVYLQMNSLKPEDTAVYYCAAGGPPAYYRYDTMDYWG QGTTVTVSS

[0211] SEQ ID NO: 27 - DNA sequences of HeVsF_Nb4

[0212] CAAGTAAAACTTGAAGAATCAGGTGGTGGACTCGTTCAAGCCGGAGGCTCACTGCG

[0213] GTTGTCTTGTGCCGCATCTGGTCGAACCTTCTCAAGTTATGCTATGGGGTGGTTCCG

[0214] GCAGGCTCCAGGTAAAGAGCGGGAATTTGTCGCAGCAATTAGTTGGTCTGGTGTAA ATACTTACTACGCTGATTCAGTTAAAGGTCGATTCACCATATCACGGGATAATGCAA AAAATGCTGGCTATCTCCAAATGAACTCTCTGAAGCCTGAAGACACTGCTGTGTATT ACTGTGCCGCAGGGGAGCCCCCGGCCTACTATAGGTACGATACTATGGACTACTGG

[0215] GGTCAAGGAACCTCAGTCACCGTCTCCTCG

[0216] SEQ ID NO: 28 - Amino acid sequences of HeVsF_Nb4

[0217] QVKLEESGGGLVQAGGSLRLSCAASGRTFSSYAMGWFRQAPGKEREFVAAISWSGVNT

[0218] YYADSVKGRFTISRDNAKNAGYLQMNSLKPEDTAVYYCAAGEPPAYYRYDTMDYWG QGTSVTVSS

[0219] SEQ ID NO: 29 - DNA sequences of HeVsF_Nb5

[0220] CAGGTTCAACTCGTCGAATCCGGGGGGGGCAGCGTACAAGCCGGAGGGTCACTCAG

[0221] GTTGTCTTGTGCCGCATCTGGTCGGACCTTCTCAAGTTATGCTATGGGGTGGTTCCG

[0222] GCAGGCTCCAGGGAAAGAGCGGGAATTTGTCGCAGTAATTAGTTGGTCTGGTGGTA

[0223] GTAGTTACTATGCTGATTCAGTAAAAGGTCGATTCACTATATCACGGGATAATGCAG

[0224] AAAATACTGGCTATCTCCAAATGAACTCTCTGAAACCTGAAGATACTGCTGTGTATT

[0225] ACTGTGCCGCCGGCCTACCCTCGGCCTACTATAGGTACGATACTATGGACTACTGGG

[0226] GTCAAGGAACCTCAGTCACCGTCTCCTCG

[0227] SEQ ID NO: 30 - Amino acid sequences of HeVsF_Nb5

[0228] QVQLVESGGGSVQAGGSLRLSCAASGRTFSSYAMGWFRQAPGKEREFVAVISWSGGSS

[0229] YYADSVKGRFTISRDNAENTGYLQMNSLKPEDTAVYYCAAGLPSAYYRYDTMDYWG QGTSVTVSS

[0230] SEQ ID NO: 31 - DNA sequences of HeVsF_Nb6

[0231] CAAGTTCAGCTTGTAGAGTCAGGCGGTGGACTCGTTCAAGCCGGAGGCTCACTGCG

[0232] GTTGTCTTGTGCCGCATCTGGTCAAACCTTCTCAAGTTATGCTGTGGGGTGGTTCCG

[0233] GCAGGCTCCCGGTAAAGAGCGGGAATTTGTCGCAGCAATTAGTTGGTCTGGTGGTG

[0234] CTACTTACTATGCTGATTCAGTAAAAGGTCGATTCACTATATCACGGGATAATGCAA

[0235] AGAATACTGTCTATCTCCAAATGAAGTCTCTGAAGCCTGAAGATACTGCTGTGTATT

[0236] ACTGTGCCGCAGGGGAGCCCCCGGCCTACTATAGGTACGATACTATGGACTACTGG

[0237] GGTCAAGGAATCTCAGTCACCGTCTCCTCG

[0238] SEQ ID NO: 32 - Amino acid sequences of HeVsF_Nb6

[0239] QVQLVESGGGLVQAGGSLRLSCAASGQTFSSYAVGWFRQAPGKEREFVAAISWSGGAT

[0240] YYADSVKGRFTISRDNAKNTVYLQMKSLKPEDTAVYYCAAGEPPAYYRYDTMDYWG QGISVTVSS

[0241] Examples

[0242] Prefusion-stabilized NiV and HeV F proteins were used for nanomice immunization. A total of 15 F-specific nanobodies were isolated from the constructed phage library. A VSV pseudotyped virus assay was established and successfully applied to evaluate the neutralization of nanobodies. The nanobodies showed cross-neutralization against HeV and NiV in both pseudotyped virus and chimeric CedV assays. Engineered multimeric nanobodies exhibited better binding activity and higher neutralization potency.

[0243] The nanobodies can be evaluated in the in vivo animal model. The neutralization mechanism of nanobodies can be assessed by identifying other neutralizing epitopes using Cryo- EM.

[0244] It is understood that the disclosed method and compositions are not limited to the particular methodology, protocols, and reagents described as these can vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention which will be limited only by the appended claims.

[0245] Disclosed are materials, compositions, and components that can be used for, can be used in conjunction with, can be used in preparation for, or are products of the disclosed method and compositions. These and other materials are disclosed herein, and it is understood that when combinations, subsets, interactions, groups, etc. of these materials are disclosed that while specific reference of each various individual and collective combinations and permutation of these compounds may not be explicitly disclosed, each is specifically contemplated and described herein. For example, if a nanobody is disclosed and discussed and a number of modifications that can be made to a number of molecules including the nanobody are discussed, each and every combination and permutation of nanobody and the modifications that are possible are specifically contemplated unless specifically indicated to the contrary. Thus, if a class of molecules A, B, and C are disclosed as well as a class of molecules D, E, and F and an example of a combination molecule, A-D is disclosed, then even if each is not individually recited, each is individually and collectively contemplated. Thus, is this example, each of the combinations A-E, A-F, B-D, B-E, B-F, C-D, C-E, and C-F are specifically contemplated and should be considered disclosed from disclosure of A, B, and C; D, E, and F; and the example combination A-D. Likewise, any subset or combination of these is also specifically contemplated and disclosed. Thus, for example, the sub-group of A-E, B-F, and C-E are specifically contemplated and should be considered disclosed from disclosure of A, B, and C; D, E, and F; and the example combination A-D. Further, each of the materials, compositions, components, etc. contemplated and disclosed as above can also be specifically and independently included or excluded from any group, subgroup, list, set, etc. of such materials. These concepts apply to all aspects of this application including, but not limited to, steps in methods of making and using the disclosed compositions. Thus, if there are a variety of additional steps that can be performed it is understood that each of these additional steps can be performed with any specific embodiment or combination of embodiments of the disclosed methods, and that each such combination is specifically contemplated and should be considered disclosed.

[0246] It must be noted that as used herein and in the appended claims, the singular forms “a,” “an,” and “the” include plural reference unless the context clearly dictates otherwise. Thus, for example, reference to “a nanobody” includes a plurality of such nanobodies, reference to “the nanobody” is a reference to one or more nanobodies and equivalents thereof known to those skilled in the art, and so forth.

[0247] “Optional” or “optionally” means that the subsequently described event, circumstance, or material may or may not occur or be present, and that the description includes instances where the event, circumstance, or material occurs or is present and instances where it does not occur or is not present.

[0248] Unless the context clearly indicates otherwise, use of the word “can” indicates an option or capability of the object or condition referred to. Generally, use of “can” in this way is meant to positively state the option or capability while also leaving open that the option or capability could be absent in other forms or embodiments of the object or condition referred to. Unless the context clearly indicates otherwise, use of the word “may” indicates an option or capability of the object or condition referred to. Generally, use of “may” in this way is meant to positively state the option or capability while also leaving open that the option or capability could be absent in other forms or embodiments of the object or condition referred to. Unless the context clearly indicates otherwise, use of “may” herein does not refer to an unknown or doubtful feature of an object or condition.

[0249] Ranges can be expressed herein as from “about” one particular value, and / or to “about” another particular value. When such a range is expressed, also specifically contemplated and considered disclosed is the range from the one particular value and / or to the other particular value unless the context specifically indicates otherwise. Similarly, when values are expressed as approximations, by use of the antecedent “about,” it will be understood that the particular value forms another, specifically contemplated embodiment that should be considered disclosed unless the context specifically indicates otherwise. It will be further understood that the endpoints of each of the ranges are significant both in relation to the other endpoint, and independently of the other endpoint unless the context specifically indicates otherwise. It should be understood that all of the individual values and sub-ranges of values contained within an explicitly disclosed range are also specifically contemplated and should be considered disclosed unless the context specifically indicates otherwise. Finally, it should be understood that all ranges refer both to the recited range as a range and as a collection of individual numbers from and including the first endpoint to and including the second endpoint. In the latter case, it should be understood that any of the individual numbers can be selected as one form of the quantity, value, or feature to which the range refers. In this way, a range describes a set of numbers or values from and including the first endpoint to and including the second endpoint from which a single member of the set (i.e. a single number) can be selected as the quantity, value, or feature to which the range refers. The foregoing applies regardless of whether in particular cases some or all of these embodiments are explicitly disclosed.

[0250] Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of skill in the art to which the disclosed method and compositions belong. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present method and compositions, the particularly useful methods, devices, and materials are as described. Nothing herein is to be construed as an admission that the present invention is not entitled to antedate such disclosure by virtue of prior invention. No admission is made that any reference constitutes prior art. The discussion of references states what their authors assert, and applicants reserve the right to challenge the accuracy and pertinency of the cited documents. It will be clearly understood that, although a number of publications are referred to herein, such reference does not constitute an admission that any of these documents forms part of the common general knowledge in the art.

[0251] Although the description of materials, compositions, components, steps, techniques, etc. can include numerous options and alternatives, this should not be construed as, and is not an admission that, such options and alternatives are equivalent to each other or, in particular, are obvious alternatives.

[0252] Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the method and compositions described herein. Such equivalents are intended to be encompassed by the following claims.

Claims

CLAIMSWe claim:1 . A nanobody that neutralizes Henipavirus.

2. The nanobody of claim 1, wherein the nanobody comprises the complementaritydetermining regions of nanobody ID nanobody ID NiVsF_Nbl, nanobody ID NiVsF_Nb2, nanobody ID NiVsF_Nb3, nanobody ID NiVsF_Nb4. nanobody ID NiVsF_Nb5, nanobody ID NiVsF_Nb6, nanobody ID NiVsF_Nb7, nanobody ID NiVsF_Nb8. nanobody ID NiVsF_Nb9, nanobody ID NiVsF_NblO, nanobody ID HeVsF_Nbl, nanobody ID HeVsF_Nb2, nanobody ID HeVsF_Nb3, nanobody ID HeVsF_Nb4, nanobody ID HeVsF_Nb5, or nanobody ID HeVsF_Nb6.

3. The nanobody of claim 1 or 2, wherein the nanobody comprises the complementarity - determining regions and one or more of the framework regions of nanobody ID nanobody ID NiVsF_Nbl, nanobody ID NiVsF_Nb2, nanobody ID NiVsF_Nb3, nanobody ID NiVsF_Nb4. nanobody ID NiVsF_Nb5, nanobody ID NiVsF_Nb6, nanobody ID NiVsF_Nb7, nanobody ID NiVsF_Nb8. nanobody ID NiVsF_Nb9, nanobody ID NiVsF_NblO, nanobody ID HeVsF_Nbl, nanobody ID HeVsF_Nb2, nanobody ID HeVsF_Nb3, nanobody ID HeVsF_Nb4, nanobody ID HeVsF_Nb5, or nanobody ID HeVsF_Nb6.

4. The nanobody of any one of claims 1-3, wherein the nanobody comprises nanobody ID nanobody ID NiVsF_Nbl, nanobody ID NiVsF_Nb2, nanobody ID NiVsF_Nb3, nanobody ID NiVsF_Nb4. nanobody ID NiVsF_Nb5, nanobody ID NiVsF_Nb6, nanobody ID NiVsF_Nb7, nanobody ID NiVsF_Nb8. nanobody ID NiVsF_Nb9, nanobody ID NiVsF_NblO, nanobody ID HeVsF_Nbl, nanobody ID HeVsF_Nb2, nanobody ID HeVsF_Nb3, nanobody ID HeVsF_Nb4, nanobody ID HeVsF_Nb5, or nanobody ID HeVsF_Nb6.

5. The nanobody of any one of claims 1-4, wherein the nanobody is humanized.

6. The nanobody of any one of claims 1-5, wherein the nanobody neutralizes Hendra virus.

7. The nanobody of claim 6, wherein the nanobody is nanobody ID HeVsF_Nb2, nanobody ID HeVsF_Nb3, nanobody ID HeVsF_Nb4, nanobody ID HeVsF_Nb5, and nanobody ID HeVsF_Nb6.

8. The nanobody of any one of claims 1-5, wherein the nanobody neutralizes Nipah virus.

9. The nanobody of claim 8, wherein the nanobody is nanobody ID nanobody ID NiVsF_Nbl, nanobody ID NiVsF_Nb2, nanobody ID NiVsF_Nb3, nanobody ID NiVsF_Nb4. nanobody ID NiVsF_Nb5, nanobody ID NiVsF_Nb6, nanobody ID NiVsF_Nb7, nanobody ID NiVsF_Nb8. nanobody ID NiVsF_Nb9, and nanobody ID NiVsF_NblO.

10. A composition comprising the nanobody of any one of claims 1-9.

11. The composition of claim 10 further comprising a pharmaceutically acceptable carrier.

12. A method of treating a Henipavirus infection, comprising administering to a subject infected with Henipavirus an effective amount of the nanobody of any one of claims 1-9 or of the composition of claim 10 or 11.

13. A method of treating a Hendra virus infection, comprising administering to a subject infected with Hendra virus an effective amount of the nanobody of claim 6 or 7 or the composition of claim 10 or 11.

14. A method of treating a Nipah virus infection, comprising administering to a subject infected with Nipah virus an effective amount of the nanobody of claim 8 or 9 or the composition of claim 10 or 11.

Images