Reuse of antibodies

Abstract

The present disclosure relates to compounds and methods for reusing antibodies.

Description

[0001] Cross - Reference to Related Applications

[0002] This application claims priority to U.S. Provisional Application No. 63 / 349,411, filed on June 6, 2022.

[0003] Reference to an Electronically Submitted "Sequence Listing"

[0004] This application contains a Sequence Listing submitted electronically in XML format as SRI - 012_P190026_SL.xml and incorporated herein by reference in its entirety. The size of the XML copy is 76,415 bytes. Background of the Invention

[0005] The discovery and development of antibodies ("Abs") for therapeutic applications can take years to identify engineered antibodies with target binding and selectivity, as well as clinical safety and efficacy. However, once identified, antibodies exhibit excellent target selectivity and generally have a clean safety window and a long duration of action (up to 24 days) in humans. The antibody drug discovery process is not suitable when a rapid response to new biological or chemical threats is required; for example, in response to a bioterrorism agent, chemical or biological weapons, or a global pandemic.

[0006] Currently, there is no platform available for the discovery, development, and manufacture of therapeutic antibodies within a time frame suitable for a rapid response to new chemical and biological threats. Alternative treatment options mainly focus on isolation, either through hospitalization or quarantine of infected sites to prevent the spread of the threat agent. Therefore, there is a need for the rapid discovery and development of antibodies against new threats, from target identification to a practical therapeutic agent, within approximately six months or less. Additionally, the ability to repurpose an antibody from its original target to recognize a second / new target opens up multiple possibilities for treating any condition other than infection, including cancer. Summary of the Invention

[0007] New synthetic compounds are disclosed herein, as well as methods for using those compounds to repurpose an original antibody that originally targets a first target so that it can be redirected to a second, novel target without structural modification of the original antibody. The methods rely on the conjugation of the original antibody with a target - redirecting synthetic compound that comprises at least two molecules covalently bound either directly or through a linker, where the first molecule is capable of non - covalently binding to the variable region of the original antibody that binds to the first target, and the second molecule is capable of non - covalently binding to a different second target.

[0008] In one embodiment, the first and / or second molecule is a synthetic polymer, also referred to herein as "Techneins". The Techneins disclosed herein are novel molecules not previously described elsewhere, although several other Techneins have been previously described in the art, such as PCT / US17 / 50119 entitled "Mass Spectrometry Distinguishable Synthetic Compounds, Libraries, and Methods Thereof", PCT / US15 / 50306 entitled "Affinity Reagent and Catalyst Discovery Though Fiber-Optic Array Scanning Technology", and PCT / US21 / 55226 entitled "High Affinity Non-Natural Ligands Against Protein Targets", the entire contents of which are incorporated herein by reference.

[0009] In one embodiment, the first and second molecules or affinity agents (e.g., Technein) can be comprehensively optimized for their targets (antibodies or novel targets), as well as for optimizing the absorption, distribution, metabolism, excretion, and toxicity (ADMET) profile and pharmacokinetics and dynamics (PK / D). Technein has been shown to have significant stability and limited toxic effects in human tissues.

[0010] In one embodiment, the identified and optimized Techneins can be directly coupled to each other or coupled to each other through a linker. In one embodiment, the Techneins can be coupled to the tight binding epitopes of antibodies previously approved by the FDA with the desired adaptive immune recruitment properties (i.e., recruiting macrophages to engage and neutralize the bound target protein) and a proven safety record in human use. Co-administering the Technein epitope conjugate with an antibody that normally binds to a first target to a second target will now cause the antibody to bind the second target by binding to the epitope conjugated to the Technein that binds to the second target. In this way, the approved antibody is repurposed to bind and immunologically recruit neutralizing factors against the second target. In some embodiments, the repurposed antibody acts through various effector functions of the antibody Fc region (ADCC, ADCP, complement activation, and interaction with other receptors on cells such as Fc receptors, etc.).

[0011] The following embodiments are exemplary but non-limiting embodiments of the present disclosure and are numbered in sequence:

[0012] 1. A synthetic compound comprising at least one unit, each unit comprising:

[0013] (i) a first molecule capable of binding to an antibody or a target-binding fragment thereof; wherein the antibody is capable of binding to a first target; and

[0014] (ii) a second molecule capable of binding to a second target;

[0015] wherein the first target and the second target are different, and wherein the first molecule and the second molecule are directly covalently linked or covalently linked via a linker.

[0016] 2. The synthetic compound according to embodiment 1, wherein the first molecule comprises a first target, an epitope thereof, an antibody-binding fragment thereof or a first synthetic polymer; and the second molecule comprises a second synthetic polymer capable of binding to a second target, wherein the first and second synthetic polymers comprise amino acid monomers and / or non-amino acid monomers.

[0017] 3. The synthetic compound according to embodiment 1, wherein the first molecule is capable of non-covalently binding to one or more complementarity-determining regions (CDRs) of an antibody.

[0018] 4. The synthetic compound according to embodiment 1, wherein the antibody has been approved by one or more of the following agencies: the Federal Drug Administration (FDA), the European Medicines Agency (EMA), the Swiss Medicines Agency, the National Medical Products Administration (NMPA) and the Pharmaceuticals and Medical Devices Agency (PMDA) of Japan.

[0019] 5. The synthetic compound according to embodiment 4, wherein the antibody is selected from Trastuzumab (Herceptin), Adalimumab (Humira), Bevacizumab (Avastin), Rituximab (Rituxan / MabThera), Infliximab (Remicade) or any other antibody in Table 1.

[0020] 6. The synthetic compound according to embodiment 1, wherein the antibody is an endogenous antibody.

[0021] 7. The synthetic compound according to embodiment 6, wherein both the first target and the first molecule are α-Gal.

[0022] 8. The synthetic compound according to Embodiment 1, wherein the target is selected from proteins, sugars, carbohydrates, nucleic acids, lipids, and combinations thereof.

[0023] 9. The synthetic compound according to Embodiment 1, wherein the target is a viral target, a tumor-specific target (e.g., a tumor-associated antigen), a tissue-specific target, a cell-specific target, a bacterial target, a fungal target, and combinations thereof.

[0024] 10. The synthetic compound according to Embodiment 1, wherein the first target is human epidermal growth factor receptor 2 (HER2) and / or the second target is a viral protein (including but not limited to the SARS-CoV-2 spike protein).

[0025] 11. The synthetic compound according to Embodiment 2, wherein the amino acids include L-amino acids, D-amino acids, β-amino acids, γ-amino acids, or combinations thereof, and the non-amino acid monomers include PAM, DhqF, DhqB, DhqO, DhqY, DhqE, N-substituted glycines, triazines, pyrimidines, or combinations thereof.

[0026] 12. The synthetic compound according to Embodiment 1, wherein the linker includes polyethylene glycol (PEG), a polyglycine sequence, a peptide, an alkyl chain, cysteine, lysine, glutamine, maleimide, dibenzyloctane, or combinations thereof.

[0027] 13. The synthetic compound according to Embodiment 1, wherein at least one unit contains multiple first and / or second molecules, wherein:

[0028] i. The multiple first molecules are directly covalently bound to each other or covalently bound to each other through a linker; and / or

[0029] ii. The multiple second molecules are directly covalently bound to each other or covalently bound to each other through a linker;

[0030] Preferably, the linker includes PEG, Lys, Gln, or combinations thereof.

[0031] 14. The synthetic compound according to Embodiment 1, wherein the first molecule and / or the second molecule can bind to the antibody or the second target, respectively, with an affinity lower than 500 nM, preferably lower than 200 nM, more preferably lower than 100 nM.

[0032] 15. The synthetic compound according to Embodiment 1, wherein the first molecule and / or the second molecule do not significantly bind to any other targets and are also referred to as being specific for the first target or the second target, respectively.

[0033] 16. An conjugate comprising at least one synthetic compound antibody or an antigen-binding fragment thereof according to any one of embodiments 1 to 15, non-covalently bound thereto, wherein the conjugate is also referred to as a repurposed antibody or a repurposed fragment thereof.

[0034] 17. The conjugate according to embodiment 16, wherein the antibody comprises light and heavy chain immunoglobulin variable and constant regions.

[0035] 18. The conjugate according to embodiment 16, wherein the antibody or antigen-binding fragment thereof is selected from monoclonal antibodies, recombinantly produced antibodies, monospecific antibodies, multispecific antibodies (including bispecific antibodies), human antibodies, engineered antibodies, humanized antibodies, chimeric antibodies, immunoglobulins, synthetic antibodies, tetrameric antibodies comprising two heavy chain and two light chain molecules, antibody light chain monomers, antibody heavy chain monomers, antibody light chain dimers, antibody heavy chain dimers, antibody light chain - antibody heavy chain pairs, intracellular antibodies, antibody fusions (sometimes referred to herein as "antibody conjugates"), hetero-conjugate antibodies, single domain antibodies, monovalent antibodies, single chain antibodies or single chain Fv (scFv), camelized antibodies, affibodies, Fab, Fab', F(ab')2 and Fv fragments, disulfide-linked Fv (sdFv), anti-idiotypic (anti-Id) antibodies (including, for example, anti-anti-Id antibodies), minibodies, domain antibodies, synthetic antibodies (sometimes referred to herein as "antibody mimetics"), and antigen-binding fragments of any of the foregoing antibodies.

[0036] 19. The conjugate according to any one of embodiments 16 to 18 and / or the synthetic compound according to any one of embodiments 1 to 15, wherein the antibody capable of binding to a first molecule, the conjugate (repurposed antibody) and / or the synthetic compound is an immunotherapeutic agent.

[0037] 20. A method of repurposing an antibody or a first target-binding fragment thereof by changing the target of the antibody or its first target-binding fragment that binds to a first target to a second target, the method comprising non-covalently conjugating the antibody or its first target-binding fragment with at least one synthetic compound according to any one of embodiments 1 to 15, wherein the second target is also referred to as a repurposed target, and wherein the conjugate is also referred to as a repurposed antibody.

[0038] 21. A method of treating a disorder in a subject in need thereof with a repurposed antibody, comprising administering to the subject an effective amount of the synthetic compound (or a portion thereof) according to embodiments 1 to 15, the antibody according to any one of embodiments 1 to 15, and / or the repurposed antibody according to any one of embodiments 16 to 19.

[0039] 22. The method according to embodiment 21, wherein the disease condition is an infectious disease caused by a pathogen comprising the second target or a cancer expressing the second target.

[0040] 23. The method according to embodiment 22, wherein the pathogen is a virus (including but not limited to the SARS-CoV-2 coronavirus).

[0041] 24. The method according to any one of embodiments 20 to 23, wherein the antibody is an endogenous antibody.

[0042] 25. A method for treating a disease caused by a pathogen in a subject in need thereof, comprising:

[0043] (i) administering to the subject an antibody that binds to a first molecule of a synthetic compound according to any one of embodiments 1 to 15, or confirming that the subject carries an endogenous antibody that binds to the compound;

[0044] (ii) exposing the subject to, or the subject having been exposed to, the pathogen, an epitope of the pathogen, and / or a part of the pathogen; and

[0045] (iii) administering to the subject the synthetic compound of (i), wherein the pathogen comprises a second target of the synthetic compound.

[0046] 26. The method according to embodiment 25, wherein the pathogen causes a disease selected from viral diseases, bacterial diseases, fungal diseases, or tick-related diseases.

[0047] 27. A method for ameliorating, treating, or reducing the incidence of cancer / tumor / malignancy in a subject in need thereof, preferably, wherein the method aids or enhances the immune system to kill or eradicate cancer cells, wherein the method comprises administering to the subject a therapeutically effective amount of one or more conjugates / repurposed antibodies, synthetic compounds, or antibodies according to any one of embodiments 1 to 19, preferably antibodies that initiate an active (or effectuate a passive) immune response to destroy cancer cells.

[0048] 28. A method for ameliorating, treating, or reducing the incidence of cancer / tumor / malignancy in a subject in need thereof, preferably, wherein the method aids or enhances the immune system to kill or eradicate cancer cells, wherein the method comprises administering to the subject a therapeutically effective amount of CAR-T cells, T cells, and / or tumor-infiltrating lymphocytes that have been contacted with or bound to an immunotherapeutic compound according to any one of embodiments 1 to 19, preferably, wherein the cells initiate an active (or effectuate a passive) immune response to destroy cancer cells.

[0049] 29. A method of using a therapeutically effective amount of the conjugate / repurposed antibody, synthetic compound or antibody according to any one of embodiments 1 to 19, or an immunotherapeutic cell conjugated or contacted with the compound or conjugate according to any one of claims 1 to 19 (including using autologous and / or heterologous cells or immortalized cell lines), in vivo or ex vivo as adoptive immunotherapy for treating cancer in a subject in need thereof, comprising administering or giving to the subject one or more conjugates / repurposed antibodies, synthetic compounds, antibodies according to any one of embodiments 1 to 19 and / or contacting the immunotherapeutic cell with one or more conjugates / repurposed antibodies, synthetic compounds, antibody cells and / or nucleic acids according to any one of embodiments 1 to 19.

[0050] 30. A composition comprising one or more conjugates / repurposed antibodies, synthetic compounds and / or antibodies according to any one of embodiments 1 to 19, and / or CAR-T cells, T cells or TILs conjugated or contacted with the compound according to any one of embodiments 1 to 19, preferably wherein the composition is a pharmaceutical composition.

[0051] 31. An immunotherapeutic compound according to any one of embodiments 1 to 19 or a composition according to embodiment 30 for cancer immunotherapy, wherein the immunotherapeutic compound or composition comprises a repurposed antibody, a fragment thereof and / or a synthetic compound of the present disclosure, and / or CAR-T cells, T cells and / or tumor infiltrating lymphocytes conjugated or contacted with the compound according to any one of embodiments 1 to 19.

[0052] 32. Use of the synthetic compound according to any one of embodiments 1 to 15, the conjugate according to any one of embodiments 16 to 19, a composition comprising the same, and / or CAR-T cells, T cells or TILs conjugated or contacted with the compound according to any one of embodiments 1 to 19 in the treatment of a disease or disorder.

[0053] 33. The use according to claim 32, wherein the disease or disorder is an infectious disease or disorder or cancer, or any other disease disorder described in the specification.

[0054] 34. Use of the synthetic compound according to any one of embodiments 1 to 15, the conjugate according to any one of embodiments 16 to 19, a composition comprising the same, and / or CAR-T cells, T cells or TILs conjugated or contacted with the compound according to any one of embodiments 1 to 19 in the manufacture of a medicament for treating a disease or disorder, preferably wherein the disease or disorder is an infectious disease or disorder or cancer, or any other disease disorder described in the specification. BRIEF DESCRIPTION OF THE DRAWINGS

[0055] Figure 1 : Examples of antibody repurposing.

[0056] Figure 2A and 2B are examples of "second molecules" containing synthetic polymers: Technein hits against the spike receptor-binding domain (SRBD) of SARS-CoV-2 (16838-89-XX); Figure 2C : Technein hits against the SRBD of Sars-Cov-2 – biotinylated hits; Figure 2D : Building blocks of the hits (all hits are from the DHQ10 library).

[0057] Figure 3A (Examples of "first molecules" containing synthetic polymers): Technein hits against Herceptin (16989-3-XX) (from the DAAP2 library) (examples of "first molecules" containing synthetic polymers); Figure 3B : Technein hits against Herceptin (16910-59-XX) (from the DAAP2 library) and Technein hits against Herceptin (16989-11-XX) (from the DHQ11 library) (examples of "first molecules" containing synthetic polymers); Figure 3C : Building blocks of the hits from the DAAP2 library; Figure 3D : Building blocks of the hits from the DHQ11 library.

[0058] Figure 4 : Repurposing agent for anti-biotin antibody against the spike protein (Technein-linker-biotin)

[0059] Figure 5A , 5B , 5C, 5D: Repurposing agents for anti-αGal antibody against the spike protein.

[0060] Figure 6A , 6B : Repurposing agent for anti-Herceptin antibody against the spike protein.

[0061] Figure 7 : Repurposing biotin antibody to bind to the SARS-2 spike protein.

[0062] Figure 8 : Biotin antibody promotes immune response in a cell function assay for clearing SARS-2 spike protein by ADCP (antibody-dependent cell phagocytosis).

[0063] Figure 9: Repurposing of Herceptin (an example of an FDA-approved antibody drug).

[0064] Figure 10 : Repurposing of α-Gal antibody (an example of an endogenous antibody).

[0065] Figure 11 : In a sandwich enzyme-linked immunosorbent assay (ELISA), the antibody repurposing Techneins bind to an anti-α-Gal antibody and a SARS-Cov2 antigen (SARS-Cov2 SRBD or Omicron S1 / S2 ECD).

[0066] Figure 12A , 12B , 12C: Performing an ADCP assay using the repurposing agent αGal-89-8. The total phagocytosis score is determined by multiplying the percentage of cells with beads by their MFI. Figure 12A and 12B : Phagocytosis score; Figure 12C : Antigen-specific phagocytosis score.

[0067] Figure 13A : Spike binding of Technein monomer and dimer; Figure 13B : Technein dimer, where two identical Techneins each bind to their own PEG molecule and then polymerize into a single PEG through a short linker. The latter PEG binds biotin.

[0068] Figure 14 : Examples of the application of antibody repurposing in vaccination against infectious agents. Detailed Description

[0069] Definitions

[0070] To better understand the present disclosure, certain terms are first defined hereinbelow. Additional definitions of the following terms and other terms are set forth throughout the specification.

[0071] As used in this specification and the appended claims, the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise.

[0072] Unless otherwise specified or obvious from the context, as used herein, the term "or" shall be understood to be inclusive and encompass both "or" and "and".

[0073] As used herein, the term "and / or" shall be taken as specifically disclosing each of the two features or components, with or without the other. Thus, the term "and / or" as used in a phrase such as "A and / or B" is intended to include A and B; A or B; A (alone) and B (alone). Similarly, the term "and / or" as used in a phrase such as "A, B, and / or C" is intended to cover each of the following aspects: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone).

[0074] As used herein, the terms "for example" and "i.e." are used only by way of illustration and not limitation, and should not be construed to refer only to those items specifically recited in the specification.

[0075] Terms such as "or more", "at least", "more than", etc., such as "at least one", should be understood to include, but not be limited to, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 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, 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, 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, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149 or 150, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 2000, 3000, 4000, 5000 or more than the values shown. Also included are any larger numbers or fractions therebetween.

[0076] Conversely, the term "not more than" includes each value less than the indicated value. In one embodiment, "not more than 100 monomers" includes 100, 99, 98, 97, 96, 95, 94, 93, 92, 91, 90, 89, 88, 87, 86, 85, 84, 83, 82, 81, 80, 79, 78, 77, 76, 75, 74, 73, 72, 71, 70, 69, 68, 67, 66, 65, 64, 63, 62, 61, 60, 59, 58, 57, 56, 55, 54, 53, 52, 51, 50, 49, 48, 47, 46, 45, 44, 43, 42, 41, 40, 39, 38, 37, 36, 35, 34, 33, 32, 31, 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, and 0 monomers. Also included are any smaller numbers or fractions therebetween.

[0077] The terms "a plurality", "at least two", "two or more", "at least second", etc., shall be understood to include, but not be limited to, at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 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, 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, 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, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149 or 150, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 2000, 3000, 4000, 5000 or more. Also included are any larger numbers or fractions therebetween.

[0078] Throughout the specification, the word "comprising" or variations thereof such as "comprises" or "comprising" shall be understood to mean including the recited element, integer, or step, or group of elements, integers, or steps, but not excluding any other element, integer, or step, or group of elements, integers, or steps. It should be understood that where an aspect is described in terms of "comprising", equivalent aspects described in terms of "consisting of" and / or "consisting essentially of" are also provided. The term "consisting of" is a closed term and excludes any element, step, or ingredient not specified in the claim. In re Gray, 53 F.2d 520, 11 USPQ 255 (CCPA 1931); Ex parte Davis, 80 USPQ 448, 450 (Bd. App. 1948) ("consisting of" is defined as "limiting the claim to the inclusion of materials other than those recited, but excluding impurities normally associated therewith"). The term "consisting essentially of" limits the scope of the claim to the specified materials or steps and those "which do not materially affect the basic and novel characteristics" of the claimed disclosure.

[0079] Unless otherwise specified or apparent from the context, as used herein, the term "about" refers to a value or composition within an acceptable error range of a particular value or composition as determined by one of ordinary skill in the art, which will depend on how the value or composition is measured or determined, i.e., the limitations of the measuring system. In one embodiment, in accordance with the practice in the art, "about" or "approximately" can refer to one or more standard deviations. "About" or "approximately" can represent a range of up to 10% (i.e., ±10%). Thus, "about" can be understood to be within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05%, 0.01%, or 0.001% greater than or less than the stated value. In one embodiment, about 5 mg can include any amount from 4.5 mg to 5.5 mg. Additionally, especially with respect to biological systems or processes, these terms can represent up to one order of magnitude or up to 5 times a numerical value. When a specific value or composition is provided in the present disclosure, unless otherwise stated, it should be assumed that the meaning of "about" or "approximately" is within the acceptable error range of that specific value or composition.

[0080] As described herein, unless otherwise specified, any concentration range, percentage range, ratio range, or integer range shall be understood to include any integer value within the recited range and, where appropriate, fractions thereof (such as one-tenth and one-hundredth of an integer).

[0081] The units, prefixes, and symbols used herein are provided in their internationally System of Units (SI)-accepted form. Numerical ranges include the values defining the range.

[0082] Unless otherwise defined, all technical terms used herein have the same ordinary meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. In one embodiment, Juo, “The Concise Dictionary of Biomedicine and Molecular Biology,” 2nd Edition, (2001), CRC Press; “The Dictionary of Cell & Molecular Biology,” 5th Edition, (2013), Academic Press; and “Oxford Dictionary Of Biochemistry And Molecular Biology,” Cammack et al., eds., 2nd Edition, (2006), Oxford University Press, provide a common dictionary of many of the terms used in this disclosure to those skilled in the art.

[0083] A “therapeutically effective amount,” “effective dose,” “effective amount,” or “therapeutically effective dose” of a therapeutic agent (e.g., Technein, a repurposed antibody, a small molecule, as described in the specification) is any amount of the agent that, when used alone or in combination with another therapeutic agent, protects a subject against the onset of a disease or promotes the regression of a disease, as evidenced by a decrease in the severity of the symptoms of the disease, an increase in the frequency and duration of asymptomatic periods of the disease, or the prevention of damage or disability caused by the affliction of the disease. These terms may be used interchangeably. A variety of methods known to those of skill in the art can be used to evaluate the ability of a therapeutic agent to promote the regression of a disease, such as in human subjects during clinical trials, in animal model systems predictive of efficacy in humans, or by determining the activity of the agent in in vitro assays. The therapeutically effective amounts and dosing regimens can be determined empirically by testing in known in vitro or in vivo (e.g., animal model) systems.

[0084] In certain embodiments, a “therapeutically effective amount” in the context of SARS CoV-2 infection is an amount sufficient to reduce one or more of the following steps of the SARS-CoV-2 life cycle: docking of viral particles with cells, introduction of viral genetic information into cells, expression of viral proteins, translation of viral RNA, transcription of viral RNA, replication of viral RNA, synthesis of new viral RNA, production of new viral particles, and release of viral particles from cells. Such a reduction in any of the foregoing may be at least 5%, preferably at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% or 100%. In some embodiments, a “therapeutically effective amount” in the context of SARS-CoV-2 infection reduces viral replication, propagation or spread by at least 5%, preferably at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% or 100%. In some embodiments, a “therapeutically effective amount” in the context of SARS-CoV-2 infection increases the survival rate of an infected subject by at least 5%, preferably at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% or 100%. In each of the foregoing, when a reduction or increase is set forth, such reduction or increase may be determined for a subject diagnosed with SARS-CoV-2 infection who has not been treated with the antibodies of the present disclosure.

[0085] The term “combination” refers to a fixed combination in unit dosage form, or the compounds of the present disclosure and combination partners (e.g., another drug as explained below, also referred to as a “therapeutic agent” or “reagent”) can be administered simultaneously independently or separately at intervals of time, particularly when these intervals of time allow the combination partners to exhibit a synergistic effect (e.g., a synergistic effect). The individual components may be packaged as a kit or separately packaged. One or both components (e.g., powder or liquid) may be reconstituted or diluted to the desired dose prior to administration. The terms “co-administered” or “administered in combination” as used herein are intended to cover the administration of selected combination partners to a single subject (e.g., a patient) in need thereof, and are intended to include treatment regimens in which the reagents need not be administered by the same route of administration or simultaneously.

[0086] As used herein, "patient" or "subject" includes any person suffering from a heart disease or disorder. The terms "subject" and "patient" are used interchangeably herein.

[0087] As used herein, the term "epitope" refers to an antigenic determinant (paratope) that interacts with (is bound by) a specific antigen-binding site in the variable region of an antibody molecule. A single antigen (such as, but not limited to, a polypeptide) can have more than one epitope. Thus, different antibodies can bind to different epitopes on an antigen and can have different biological effects depending on the epitope to which they bind. The term "epitope" also refers to the site on an antigen at which a B-cell and / or T-cell response occurs. It also refers to the region of an antigen to which an antibody binds. An epitope can be defined as a structural epitope (the part of the antigenic determinant that contacts the CDR loops of an antibody) or a functional epitope (a subset of the structural epitope that includes those energetic residues located at the center of the structural epitope and that directly contribute to the affinity of the antibody-epitope interaction). An antigenic epitope can have immunological activity (cryptotope) after antigen fragmentation or denaturation. An epitope can be linear or conformational (comprising non-linear amino acids that are clustered together in a folded three-dimensional structure). An epitope can include groups of surface molecules with chemical activity (such as amino acids, sugar side chains, phosphoryl or sulfonyl groups), and an epitope can have specific three-dimensional structural features and / or specific charge characteristics. An epitope typically includes at least 3 to 15 amino acids.

[0088] The term "antibody" is used herein in the broadest sense and encompasses various antibody structures and antibody fragments, provided they exhibit the desired antigen-binding activity and fusion proteins comprising an antibody, as well as any other modified configurations of immunoglobulin molecules containing an antigen recognition site. Antibodies include antibodies of any class, such as IgG, IgA or IgM (or their subclasses), and antibodies need not belong to any particular class. Immunoglobulins can be classified into different classes based on the amino acid sequence of the constant domain of their heavy chains. There are five main classes of immunoglobulins: IgA, IgD, IgE, IgG and IgM, several of which can be further divided into multiple subclasses (subtypes, also known as isotypes), such as IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2. The constant regions of the heavy chains corresponding to different classes of immunoglobulins are called α, δ, ε, γ and μ, respectively. The subunit structures and three-dimensional configurations of different classes of immunoglobulins are well known. Examples of antibody structures include, but are not limited to, monoclonal antibodies, recombinantly produced antibodies, monospecific antibodies, multispecific antibodies (including bispecific antibodies), human antibodies, engineered antibodies, humanized antibodies, chimeric antibodies, immunoglobulins, synthetic antibodies, tetrameric antibodies comprising two heavy chain and two light chain molecules, antibody light chain monomers, antibody heavy chain monomers, antibody light chain dimers, antibody heavy chain dimers, antibody light chain - antibody heavy chain pairs, intracellular antibodies, antibody fusions (sometimes referred to herein as "antibody conjugates"), hetero-conjugate antibodies, single domain antibodies, monovalent antibodies, single chain antibodies or single chain Fv (scFv), camelized antibodies, affibodies, Fab, Fab', F(ab')2 and Fv fragments, disulfide-linked Fv (sdFv), anti-idiotypic (anti-Id) antibodies (including, for example, anti-anti-Id antibodies), minibodies, domain antibodies, synthetic antibodies (sometimes referred to herein as "antibody mimetics"). In one embodiment, the antibody has been previously approved by a regulatory agency for clinical use.

[0089] A "humanized" antibody refers to a chimeric antibody that contains amino acid residues from non-human HVRs and amino acid residues from human FRs. In certain embodiments, a humanized antibody will comprise at least one, and typically two, variable regions in which all or substantially all of the HVRs (e.g., CDRs) correspond to the corresponding portions of a non-human antibody, and all or substantially all of the FRs correspond to the corresponding portions of a human antibody. A humanized antibody optionally comprises at least a portion of an antibody constant region derived from a human antibody. The "humanized form" of an antibody, such as a non-human antibody, refers to the antibody that has been humanized.

[0090] The term "variable region" or "variable domain" refers to the heavy or light chain domain of an antibody that is involved in binding of the antibody to an antigen. The variable domains of the heavy and light chains of a native antibody (VH and VL, respectively) generally have similar structures, each domain comprising four conserved framework regions (FRs) and three hypervariable regions (HVRs). A single VH or V L The domain may be sufficient to confer antigen-binding specificity. In addition, VH or VL domains from antibodies that bind to an antigen can be used to isolate antibodies that bind to a specific antigen to screen libraries of complementary VL or VH domains, respectively. See, e.g., Portolano et al., J. Immunol. 150:880-887 (1993); Clarkson et al., Nature 352:624-628 (1991).

[0091] The term "antigen-binding fragment" refers to a molecule other than an intact antibody that contains a portion of an intact antibody and is capable of binding to an antigen to which the intact antibody binds. Examples of antigen-binding fragments include, but are not limited to, Fv, Fab, Fab', Fab'-SH, F(ab')2, diabodies, dAb, linear antibodies, single-chain antibodies (e.g., scFv); single-domain antibodies; antigen-binding fragments of bivalent or bispecific antibodies; camelized antibodies; single-domain antibodies, maxibodies, minibodies, nanobodies, intracellular antibodies, diabodies, triabodies, tetra-bodies, v-NAR, and bis-scFv, and other fragments having the ability to bind to a desired antigen (e.g., the SARS-COV spike).

[0092] The term "antigen-binding site" refers to the portion of an antibody molecule that contains determinants that form an epitope interface capable of binding to a polypeptide or its epitope. In the case of a protein (or protein mimetic), the antigen-binding site typically contains one or more loops (e.g., loops of at least 4 amino acids or amino acid mimetics) that form an interface capable of binding to a polypeptide. Generally, the antigen-binding site of an antibody molecule contains at least one or two CDRs and / or hypervariable loops, or more typically at least three, four, five, or six CDRs and / or hypervariable loops.

[0093] As used herein, the term "CDR" refers to the complementarity determining regions within the variable sequences of an antibody. Each of the variable regions of the heavy and light chains has three CDRs, which are CDR1, CDR2, and CDR3 of each variable region. As used herein, the term "CDR set" refers to the set of three CDRs that occur within a single variable region and that are capable of binding an antigen. The exact boundaries of these CDRs are defined differently according to different systems. The system described by Kabat (Kabat et al., Sequences of Proteins of Immunological Interest (U.S. National Institutes of Health, Bethesda, MD (1987) and (1991))) provides not only a defined residue numbering system applicable to any variable region of an antibody, but also the precise residue boundaries that define the three CDRs. These CDRs may be referred to as Kabat CDRs. Subparts of the CDRs may be designated as L1, L2, and L3 or H1, H2, and H3, where "L" and "H" denote the light and heavy chain regions, respectively. These regions may be referred to as Chothia CDRs, the boundaries of which overlap with the Kabat CDRs. Padlan (FASEB J. 9:133-139 (1995)) and MacCallum (J Mol Biol. 262(5):732-45 (1996)) have described other boundaries that define CDRs that overlap with the Kabat CDRs. There are other CDR boundary definitions that may not strictly follow one of the above systems, but will still overlap with the Kabat CDRs, and although specific residues or groups of residues or even entire CDRs may not significantly affect antigen binding according to prediction or experimental results, they may be shortened or lengthened. The methods used herein may utilize CDRs defined according to any of these systems, although preferred embodiments use Kabat- or Chothia-defined CDRs.

[0094] As used herein, the term "specifically binds" refers to the ability of a molecule to bind to a binding partner with a degree of affinity or avidity such that the molecule can be used to distinguish the binding partner from a suitable control in a binding assay or other binding situation. For an antibody, the term "specifically binds" refers to the ability of the antibody to bind a particular antigen with a degree of affinity or avidity such that, compared to a suitable reference antigen, the antibody can be used to distinguish the particular antigen from other antigens, e.g., to preferentially target certain cells (such as muscle cells) by binding to the antigen, as described herein. In some embodiments, preferably measured by the method of the examples (Biolayer Interferometry (BLI)), if the KD (affinity) of the antibody for the target is at least about 10 -4 M, 10 -5 M, 10-6 M, 10 -7 M, 10 -8 M, 10 -9 M, 10 -10 M, 10 -11 M, 10 -12 M, 10 -13 If M or less, then the antibody can specifically bind to the target.

[0095] The term "antibody repurposing" refers to the change of the original target (binding molecule) of an antibody from a first / original target to a second / new target. Antibody repurposing can be achieved by conjugating the original antibody with a synthetic compound comprising (i) a first molecule that can bind to the variable region of the original antibody and (ii) a second molecule that can bind to the second / new target, wherein the first molecule and the second molecule are directly covalently linked or covalently linked through a linker. The synthetic compound acts as a bridge between the original antibody and its new target, thereby repurposing the antibody for the new target. The second molecule comprises a synthetic polymer, herein referred to as Technein.

[0096] The term "Technein" refers to a specific subset of synthetic polymers composed of covalently linked amino acid monomers and non-amino acid monomers. The amino acids can be, for example, D-amino acids (e.g., D-Glu(tBu); D-Lys), L-amino acids, β-amino acids, γ-amino acids, or combinations thereof. Non-limiting examples of monomers useful for Technein are shown in the figure and include PAM, DhqF, DhqB, DhqO, DhqY, DhqE, N-substituted glycines, triazines, pyrimidines, and combinations thereof.

[0097] The term "linker" refers to a chemical moiety comprising a covalent bond or chain of atoms that covalently attaches one molecule to another (e.g., Technein to another Technein). In various embodiments, the linker includes divalent groups such as alkyldiyl, aryldiyl, heteroaryldiyl, repeating units of moieties such as --(CR2)nO(CR2)n--, alkoxy (e.g., polyoxyethylene, PEG, polymethyleneoxy), and alkylamino (e.g., polyaminoethyl); and diesters and amides, including succinate, succinamide, diglycolate, malonate, and hexamide. In various embodiments, the linker can comprise one or more amino acid residues such as valine, phenylalanine, cysteine, lysine, and homoarginine.

[0098] The term "synthetic" is generally used herein to refer to compounds or molecules that are not naturally occurring, such as the compounds described herein.

[0099] As used herein, the term "polypeptide" refers to a polymer of amino acids. The polymer can be a linear or branched polymer, can contain modified amino acids, and can be interspersed with non-amino acids. The term also encompasses modified amino acid polymers; for example, disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, or any other manipulation, such as conjugation with a labeled component. Polypeptides can be isolated from natural sources, produced by prokaryotic or eukaryotic hosts using recombinant techniques, or can be the products of synthetic methods. In some embodiments, the polypeptide is greater than 50 amino acids in length.

[0100] As used herein, a "peptide" is less than or equal to 50 amino acids in length, for example, about 5, 10, 15, 20, 25, 30, 35, 40, 45, or 50 amino acids in length.

[0101] As used herein, the terms "Her-2", "ErbB2", "c-Erb-B2", "HER2", "Her2", and "neu" are used interchangeably and refer to native HER2 and its allelic variants. Unless otherwise specified, the terms "HER2", "ErbB2", "c-Erb-B2", "HER2", and "Her2" as used herein refer to the human protein. The gene encoding Her2 is referred to herein as "ErbB2".

[0102] The term "aptamer" refers to a biomolecule that can be designed or selected to bind tightly to other ligands, for example using a technique called systematic evolution of ligands by exponential enrichment (SELEX; Tuerk C, Gold L: "Systematic evolution of ligands by exponential enrichment: RNA ligands to bacteriophage T4 DNA polymerase." Science 1990, 249: 505-510). Aptamers can be peptides. Aptamers can also be nucleic acids. For example, nucleic acid aptamers can be selected from pools of random sequence oligonucleotides that have high binding affinity and specificity for a wide range of biomedically relevant targets, suggesting that aptamers have broad therapeutic applications (Keefe, Anthony D., Supriya Pai and Andrew Ellington. "Aptamers as therapeutics." Nature Reviews Drug Discovery 9.7 (2010): 537-550). These characteristics also suggest that aptamers can be widely used as drug delivery vehicles (Levy-Nissenbaum, Etgar et al. "Nanotechnology and aptamers: applications in drug delivery." Trends in biotechnology 26.8 (2008): 442-449; and Hicke B J, Stephens A W. "Escort aptamers: a delivery service for diagnosis and therapy." J Clin Invest 2000, 106: 923-928). Aptamers can also be constructed to act as molecular switches, changing properties in response to queries, such as RNA aptamers that bind fluorophores to mimic green fluorescent protein activity (Paige, Jeremy S., Karen Y. Wu and Samie R. Jaffrey. "RNA mimics of green fluorescent protein." Science 333.6042 (2011): 642-646).It has also been proposed that aptamers can be used as components of a targeted siRNA therapeutic delivery system, such as targeting cell surface proteins (Zhou, Jiehua and John J. Rossi. "Aptamer-targeted cell-specific RNA interference." Silence 1.1 (2010): 4). In some embodiments, an "aptamer" refers to single-stranded or double-stranded oligodeoxyribonucleic acid (oligoDNA), oligoribonucleic acid (oligoRNA), or oligoDNA / RNA, or any analog thereof, that can specifically bind to a target molecule, such as a peptide. Advantageously, aptamers exhibit a relatively high degree of specificity and affinity for their targets. The generation of aptamers is described in U.S. Patent No. 5,270,163; Ellington & Szostak 1990 (Nature 346:818-822); Tuerk & Gold 1990 (Science 249:505-510); or "The Aptamer Handbook: Functional Oligonucleotides and Their Applications", edited by Klussmann, Wiley-VCH 2006, ISBN 3527310592, which is incorporated herein by reference. The term "photoaptamer" refers to an aptamer that contains one or more photoreactive functional groups that can covalently bind to or crosslink with a target molecule. The term "spiegelmer" refers to an aptamer that includes L-DNA, L-RNA, or other left-handed nucleotide derivatives or nucleotide-like molecules. Aptamers containing left-handed nucleotides are resistant to degradation by naturally occurring enzymes that typically act on substrates containing right-handed nucleotides. The term "peptide mimetic" refers to a non-peptide agent that is a topological analog of the corresponding peptide. Methods for rationally designing peptide mimetics of peptides are known in the art. For example, the rational design of three peptide mimetics based on the sulfated octamer peptide CCK26-33, and the rational design of two peptide mimetics based on the 11-mer peptide substance P, and related peptide mimetic design principles are described in Horwell 1995 (Trends Biotechnol 13:132-134).

[0103] As used herein, the term "affinity" refers to the strength of the sum of non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). Unless otherwise stated, as used herein, "binding affinity" refers to the intrinsic binding affinity that reflects the 1:1 interaction between the members of a binding pair (e.g., an antibody and an antigen). The affinity of a molecule X for its partner Y is typically expressed by the dissociation constant (KD). Methods for determining binding affinity are known in the art and include surface plasmon resonance (e.g., SPR, BIACORE) or similar techniques (e.g., ForteBio; Biolayer Interferometry (BLI)).

[0104] The term "immunotherapeutic agent" refers to the synthetic compounds or repurposed antibodies (conjugates) of the present disclosure, which can be understood as immunotherapeutic compounds or compositions.

[0105] Repurposing of Antibodies

[0106] The present disclosure relates to a method for repurposing an antibody by altering its target without covalently modifying its original structure. In one embodiment, the method allows for the repurposing of antibodies that have been approved by health regulatory agencies such as the US Food and Drug Administration (FDA), the European Medicines Agency (EMA), the Swissmedic, the National Medical Products Administration (NMPA), and the Pharmaceuticals and Medical Devices Agency (PMDA) of Japan, without the need for the extensive clinical testing to the same extent as that applied to newly developed antibodies prior to approval. The repurposing of an antibody is achieved by altering its target. Any antibody has an original target, i.e., the original molecule that the antibody is designed or produced to bind through the complementarity-determining regions (CDRs) in its variable region. The present disclosure provides a method for altering the original target by non-covalently conjugating the antibody with a synthetic compound that serves as a bridge between the variable region of the antibody (which can bind to the original target) and a second new target. Such a synthetic compound or bridge is capable of binding to the variable region of the antibody at one end and to the second / new target at the other end. To achieve this, the synthetic compound comprises at least two different molecules that are directly covalently linked or covalently linked through a linker. One molecule ("the first molecule") is capable of binding to the variable region of the antibody. The other molecule ("the second molecule") is capable of binding to the second / new target. At least one of these molecules is a synthetic polymer, which in some embodiments may be referred to herein as a "Technein". Figure 1 Examples of antibody repurposing are visible. Antibody repurposing can alter the clinical use of an antibody without covalent modification. In one embodiment, an antibody initially approved by a health regulatory agency for cancer treatment can be repurposed as a vaccine against an infectious agent. In addition, CAR-T cells can also be repurposed by altering their targets by the methods disclosed herein.

[0107] Antibody repurposing takes advantage of the immunomodulatory efficacy and safety of previously FDA-approved antibodies that took decades to develop and validate for clinical use, and enables the discovery and development of affinity targeting agents within months to effectively repurpose the antibody to any desired target. This approach allows for the rapid availability of antibody-based countermeasures without the time and cost of discovery and development, and importantly, since the antibody has been GMP manufactured for its primary approved indication, the lead time for establishing antibody GMP production at a scale suitable for at-risk and exposed populations can be avoided. In contrast, Technein-epitope conjugates are relatively easy to manufacture, and a general GMP validation process can be pre-established for rapid response. As described above, this approach can be extended beyond Technein to include any target affinity molecule related to a synthetic epitope or hapten of an antibody. This can include small molecules, peptides, scFv, nucleic acids, and aptamers as well as Technein. In some embodiments, the Technein is selected from those in Figures 2 and 3. In some embodiments, the Technein is selected from the Technein described in PCT / US17 / 50119 entitled "MASS SPECTROMETRY DISTINGUISHABLE SYNTHETIC COMPOUNDS, LIBRARIES, AND METHODS THEREOF", PCT / US15 / 50306 entitled "Affinity Reagent and Catalyst Discovery Though Fiber-Optic Array Scanning Technology", and PCT / US21 / 55226 entitled "HIGH AFFINITY NON-NATURAL LIGANDS AGAINST PROTEIN TARGETS". The content of which is incorporated herein by reference in its entirety.

[0108] Synthetic Compounds for Antibody Repurposing

[0109] In one embodiment, the present disclosure provides a method of repurposing an antibody. In one embodiment, the method involves non-covalently conjugating an original antibody with a synthetic compound that can act as a bridge between the original antibody and a new target of the repurposed antibody. In one embodiment, the antibody and the synthetic compound are covalently linked. In one embodiment, the synthetic compound comprises the following moieties:

[0110] (i) A first molecule capable of binding to an antibody or a target-binding fragment thereof; wherein the antibody is capable of binding to a first target; and

[0111] (ii) A second molecule capable of binding to a second target;

[0112] wherein the first target and the second target are different, and wherein the first molecule and the second molecule are directly covalently linked or covalently linked via a linker.

[0113] In one embodiment, the first molecule is capable of binding to the variable region of the antibody. In some embodiments, the first molecule comprises the first target or a fragment thereof. In some embodiments, the first molecule comprises an epitope of an antibody to be repurposed. In one embodiment, the first molecule is selected from any of the targets of the antibodies described in Tables 1 to 3, or an antibody-binding fragment thereof. In some embodiments, the antibody is an endogenous antibody and the first molecule is selected from its natural targets. In some embodiments, the first molecule is α-Gal. In some embodiments, the first molecule is biotin.

[0114] In some embodiments, the first molecule comprises a synthetic polymer. In some embodiments, the synthetic polymer comprises amino acid monomers and non-amino acid monomers. In some embodiments, the synthetic polymers, amino acids, and monomers are selected from those in Figure 3. In some embodiments, the synthetic polymer is also referred to as Techneins. In some embodiments, the Technein is selected from the Techneins described in PCT / US17 / 50119 entitled "MASS SPECTROMETRY DISTINGUISHABLE SYNTHETIC COMPOUNDS, LIBRARIES, AND METHODS THEREOF", PCT / US15 / 50306 entitled "Affinity Reagent and Catalyst Discovery Though Fiber-Optic Array Scanning Technology", and PCT / US21 / 55226 entitled "HIGH AFFINITY NON-NATURAL LIGANDS AGAINST PROTEIN TARGETS", the entire contents of which are incorporated herein by reference in their entirety. In one embodiment, the first molecule is selected from small molecules, peptides, scFvs, nucleic acids, and aptamers.

[0115] In some embodiments, the second molecule is capable of binding to a second target. In one embodiment, the second molecule comprises a synthetic polymer. In some embodiments, the synthetic polymer comprises amino acid monomers and non-amino acid monomers. In some embodiments, the synthetic polymer, amino acids, and monomers are selected from those in Figure 2. In some embodiments, the synthetic polymer is a Technein. In one embodiment, the first molecule and / or the second molecule are selected from small molecules, peptides, scFvs, nucleic acids, and aptamers.

[0116] In one embodiment, the first molecule and the second molecule are covalently linked. In one embodiment, the first molecule and the second molecule are directly linked to each other. In one embodiment, the first molecule and the second molecule are linked to each other through a linker.

[0117] In some embodiments, the synthetic compound comprises one Technein. In some embodiments, the synthetic compound comprises two Techneins, where one is the first molecule and the other is the second molecule. Figure 1 . In some embodiments, the synthetic compound comprises the first molecule and / or the second molecule, and each molecule comprises two or more Techneins that are directly linked to each other or interconnected through a linker. In some embodiments, two Techneins in the first molecule or the second molecule are interconnected, as Figure 13B shown. In some embodiments, the first molecule comprises a Technein dimer, where each monomer is linked to a PEG molecule, and where two PEG molecules converge at a Lys-Gln linker that can be linked to a PEG, which can be linked to the second molecule.

[0118] In some embodiments, the linker comprises polyethylene glycol (PEG), PEG. In some embodiments, the linker comprises PEG, a peptide, an alkyl chain, cysteine, lysine, glutamine, maleimide, dibenzyloctane, or a combination thereof.

[0119] antibody

[0120] Any antibody can be repurposed using the methods of the present disclosure. In some embodiments, the antibody has previously been approved by the U.S. Food and Drug Administration (FDA), the European Medicines Agency (EMA), the Swiss Medicines Agency, the National Medical Products Administration (NMPA), and the Pharmaceuticals and Medical Devices Agency (PMDA) of Japan, or any other health agency responsible for antibody approval. In some embodiments, the antibody is an endogenous antibody (endogenous to the subject to be treated with the repurposed antibody). In some embodiments, the endogenous antibody is recombinantly expressed and binds to the synthetic compound in vitro.

[0121] Examples of previously approved antibodies can be found on the websites of health agencies. In some embodiments, the antibody is selected from those in Table 1.

[0122] Table 1: Exemplary Antibodies for Repurposing

[0123]

[0124]

[0125]

[0126]

[0127]

[0128]

[0129]

[0130]

[0131]

[0132]

[0133]

[0134]

[0135]

[0136]

[0137]

[0138]

[0139] In some embodiments, the antibody is selected from trastuzumab (Herceptin), adalimumab (Humira), bevacizumab (Avastin), rituximab (Rituxan / MabThera), infliximab (Infliximab), and (Remicade). In some embodiments, the antibody is Herceptin. In some embodiments, the antibody is selected from endogenous anti-α-galactosidase (Gal) antibodies. Alpha-Gal may also be referred to as a-Gal and α-Gal herein and in the art.

[0140] The mechanism of action of chimeric antigen receptor T cell (CAR-T) therapy also relies on antibody-like molecules. Therefore, antibody repurposing can also be used to repurpose CAR-T cells by altering their targets in a similar manner. Examples of repurposable CAR-T cell therapies include, but are not limited to, those in Table 2.

[0141] Table 2: Examples of FDA-approved CAR-T cell therapies

[0142]

[0143]

[0144] In some embodiments, CAR-T cells are inherently targeted to CD19, CD20, or BCMA.

[0145] Table 3: Non-exhaustive list of ongoing CAR-T cell therapy studies for solid organ malignancies

[0146]

[0147]

[0148]

[0149]

[0150] Target

[0151] The first / original target and the second / new target can be any target. The first target determines the antibody, and generally, it doesn't matter what the target is, because the ability of the antibody to recognize and bind to that target will be eliminated due to the binding of the synthetic compound to its variable region. The first or original targets of some antibodies that can be used according to the present disclosure are listed in Tables 1-3, but are not limited thereto. The antibody can be recognized by any first molecule. Examples of the first molecule are as described above. When the first molecule is Technein, a Technein that can bind to the antibody must be obtained. Once the second target is selected, the subsequent step is to obtain a Technein that can specifically bind to it. In one embodiment, the Technein is identified by screening a Technein library. For some antibodies, the first molecular component of the synthetic compound of the present disclosure does not need to be Technein, but can be the natural target / ligand of the antibody, or a fragment thereof (e.g., an epitope). In one embodiment, if the antibody to be repurposed is an anti-biotin antibody, the first molecule of the synthetic compound may contain biotin. If the antibody is an anti-α-Gal antibody, the first molecule may contain α-Gal.

[0152] The second target determines the new use of the repurposed antibody. A second molecule that binds to the second target junction must be identified and can be directly linked or linked via a linker to the first molecule. In some embodiments, the second molecule is a Technein. In some embodiments, a Technein library is screened to identify those that bind to the second target.

[0153] The target can be any type of biomolecule, including, for example, simple intermediary metabolites, sugars, lipids, and hormones, as well as macromolecules such as complex carbohydrates, phospholipids, nucleic acids, and proteins. Some exemplary classes of targets include, but are not limited to, viral antigens, bacterial antigens, fungal antigens, tick antigens, protozoan and other parasite antigens, tumor antigens, antigens involved in autoimmune diseases, allergies, and transplant rejection, and various other antigens. In one embodiment, the target is selected from microbial antigens such as viral antigens, fungal antigens, or bacterial antigens; or therapeutic antigens such as antigens associated with cancer cells or growth or antigens associated with autoimmune diseases. In some embodiments, the target is selected from small molecules, nucleotides, polynucleotides, peptides, polypeptides, proteins, lipids, carbohydrates, other immunogenic molecules, and combinations thereof. Exemplary diseases, pathogens, pathogen polypeptides, and disease-related polypeptides are known, and those of ordinary skill in the art can readily identify other targets. The first target and the second target can be selected from the same molecular library. Thus, any reference to the second target can also serve as an example of the first target. The first target and the second target of the synthetic compounds of the present disclosure must be different in order to enable the repurposing of the antibody.

[0154] In some embodiments, the repurposed antibody can be used as an immunotherapeutic agent for vaccination against pathogens or for treating resulting infections, including persistent viral infections. Pathogens can include bacteria, protozoa, viruses, and prions, as well as other prion-like particles that cause diseases and disorders. Bacterial pathogens include, for example, Escherichia coli, Klebsiella, Staphylococcus, Acinetobacter, and Pseudomonas species, especially drug-resistant species and strains. Bacteria also include strains / species of Salmonella, such as Salmonella typhimurium. Viruses include the Retroviridae (e.g., HIV, including HIV fusion peptide antigen), Orthomyxoviridae, Paramyxoviridae, Arenaviridae, 5 Filoviridae, and / or Coronaviridae (e.g., SARS-CoV, SARS-CoV-2 fusion peptide, and / or PEDV).

[0155] Non-limiting examples of viral antigens that can be selected as the second target include antigens from Coronaviridae viruses (severe acute respiratory syndrome coronavirus (SARS-CoV), Middle East respiratory syndrome-related coronavirus (MERS-CoV), and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)), retroviral antigens such as retroviral antigen human immunodeficiency virus (HIV) antigens, such as gene products of gag, pol, and env genes, Nef protein, reverse transcriptase, and other HIV components; hepatitis virus antigens such as the S, M, and L proteins of hepatitis B virus, the pre-S antigen of hepatitis B virus, and other hepatitis (e.g., hepatitis A, hepatitis B, and hepatitis C) virus components such as hepatitis C virus RNA; influenza virus antigens such as hemagglutinin and neuraminidase and other influenza virus components; measles virus antigens such as the measles virus fusion protein and other measles virus components; rubella virus antigens such as proteins E1 and E2 and other rubella virus components; rotavirus antigens such as VP7sc and other rotavirus components; cytomegalovirus antigens such as envelope glycoprotein B and other cytomegalovirus antigen components; respiratory syncytial virus antigens such as RSV fusion protein, M2 protein, and other respiratory syncytial virus antigen components; herpes simplex virus antigens such as immediate early proteins, glycoprotein D, and other herpes simplex virus antigen components; varicella-zoster virus antigens such as gpI, gpII, and other varicella-zoster virus antigen components; Japanese encephalitis virus antigens such as protein E, M-E, M-E-NS1, NS1, NS1-NS2A, 80% E, and other Japanese encephalitis virus antigen components; rabies virus antigens such as rabies glycoprotein, rabies nucleoprotein, and other rabies virus antigen components. For other examples of viral antigens, see Fundamental Virology, 2nd Edition, edited by Fields, BN and Knipe, DM (Raven Press, New York, 1991).

[0156] In some embodiments, the target is a viral protein from Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV), Middle East Respiratory Syndrome-related Coronavirus (MERS-CoV), or Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). These three viruses mainly mediate viral invasion by binding of the spike protein (S protein) to host cell receptors and determine viral tissue or host tropism. The host cell receptor protein of SARS-CoV-2 is angiotensin-converting enzyme 2 (ACE2). The spike protein (S protein) can bind to the ACE2 receptor and is cleaved by host proteases into an S1 polypeptide containing the receptor-binding domain (SARS-CoV-2 RBD) and an S2 polypeptide responsible for mediating viral fusion with the cell membrane and thus invading the host. In one embodiment of the present disclosure, the coronavirus SARS-CoV-2 spike protein (SARS-CoV-2 S protein), its extracellular region, S1 subunit, or receptor-binding region is used as the target. In one embodiment, the target protein is from the viral envelope (E), membrane (M), or nucleocapsid (N). In some embodiments, the coronavirus is an SARS-CoV-2 variant selected from the group consisting of Alpha, Beta, Gamma, Delta, and Epsilon variants.

[0157] In some embodiments, the virus is a virus of the family Orthomyxoviridae. Infections with influenza viruses such as influenza A, B, and C mainly rely on two envelope glycoproteins: hemagglutinin (HA) and neuraminidase (NA), which are responsible for viral attachment and the invasion of virus particles into cells. The initiation of influenza virus infection is caused by the attachment of the hemagglutinin (HA) protein to sialic acid-containing cell receptors (glycoproteins and glycolipids) on the surface of the virion. The neuraminidase (NA) protein mediates the processing of sialic acid receptors, and viral invasion of cells depends on HA-dependent receptor-mediated endocytosis. In one embodiment of the present disclosure, the influenza A H5N1 hemagglutinin (HA) protein is used as an antigen, and the influenza B hemagglutinin protein (HA1 subunit) can also be used as a target.

[0158] In some embodiments, the virus is a virus of the family Filoviridae, and its representatives can be Ebola virus of the genus Ebola and Marburg virus of the genus Marburg. The only protein present on the surface of the Ebola virus is the glycoprotein (GP). The GP1,2 trimer that forms the viral surface spike contains two subunits, GP1 and GP2, linked by disulfide bonds. It is known that GP1 mediates viral attachment to host cells, while GP2 is involved in membrane fusion. In one embodiment, the Ebola virus glycoprotein (GP) is selected as the target, such as the extracellular domain of GP, the subunit GP proteins (GP1 and / or GP2).

[0159] In some embodiments, the virus is a Flaviviridae virus, which mainly includes the genera Flavivirus, Pestivirus, Pegivirus, and Hepatitis C virus. Among them, the Flaviviridae virus includes Zika virus (ZIKV), Dengue virus (DV), West Nile virus, Japanese encephalitis virus, and Yellow fever virus. The genus Hepatitis C virus includes Hepatitis C virus (HCV). The flavivirus envelope protein plays an important role in the infection of host cells by the virus and mediates the entry of the virus into host cells. It consists of three independent structural envelope domains I, II, and III (EDI, EDII, and EDIII). EDI is the structural core domain of the envelope protein, which can stabilize the overall conformation of the protein, and the glycosylation sites in EDI are related to virus production, pH sensitivity, and neuroinvasiveness. Due to the immunological advantages of the fusion loop epitope and the envelope dimer epitope, EDII plays an important role in membrane fusion. In addition, EDIII is the main target of neutralizing antibodies. The Zika virus envelope protein (“E” or “EP”) consists of three different domains. E domain I (E-DI) is the central domain that organizes the entire E protein structure. E domain II (E-DII) is formed by two extended loops extending from E-DI and is located in the pocket region between E-DI and E domain III (E-DIII). E-DIII is an immunoglobulin-like domain, which forms small protrusions on the surface of the originally smooth, spherical, mature virus particles and is considered to interact with cell receptors on target cells. In one embodiment, the Zika virus envelope protein E-DIII is selected as the target. In one embodiment, the Hepatitis C virus envelope glycoproteins E1 and / or E2 are selected as the target.

[0160] In some embodiments, the virus is HCV. The HCV RNA genome encodes a single polyprotein, which is cleaved into three structural proteins (core protein, glycoprotein E1, and E2) and seven non-structural proteins (p7, NS2, NS3, NS4A, NS4B, NS5A, and NS5B) during translation or post-translation. The envelope proteins, glycoproteins E1 and E2, form a heterodimer and constitute the virus envelope protein, which plays an important role in mediating virus entry and morphogenesis when the virus enters host cells. The Hepatitis C virus envelope protein glycan binds to specific proteins on the surface of host hepatocytes, initiating the virus invasion process. This process involves a large number of host receptors / coreceptors. Among them, E2 is the main HCV envelope protein glycan and directly interacts with the receptor / coreceptor. For a long time, it has been believed that E1 does not directly interact with host receptors during this process, but rather initiates membrane fusion with E2 by maintaining the functional E2 conformation required for receptor binding. In some embodiments, the second target is E1 or E2.

[0161] Non-limiting examples of bacterial antigens that can be used as a second target include, but are not limited to, gonococcal bacterial antigens; Bordetella pertussis bacterial antigens such as pertussis toxin, filamentous hemagglutinin, pertactin, FIM2, FIM3, adenylate cyclase, and other Bordetella pertussis bacterial antigen components; Corynebacterium diphtheriae bacterial antigens such as diphtheria toxin or toxoid and other Corynebacterium diphtheriae bacterial antigen components; Clostridium tetani bacterial antigens such as tetanus toxin or toxoid and other Clostridium tetani bacterial antigen components; Streptococcus bacterial antigens such as M protein and other Streptococcus bacterial antigen components; Gram-negative bacillus bacterial antigens such as lipopolysaccharide and other Gram-negative bacterial antigen components; Mycobacterium tuberculosis bacterial antigens such as mycolic acid, heat shock protein 65 (HSP65), 30 kDa major secreted protein, antigen 85A, and other mycobacterial antigen components; Helicobacter pylori bacterial antigen components; Streptococcus pneumoniae bacterial antigens such as pneumolysin, pneumococcal capsular polysaccharide, and other Streptococcus pneumoniae bacterial antigen components; Haemophilus influenzae bacterial antigens such as capsular polysaccharide and other Haemophilus influenzae bacterial antigen components; Bacillus anthracis bacterial antigens such as Bacillus anthracis protective antigen and other Bacillus anthracis bacterial antigen components; Rickettsia bacterial antigens such as rOMPs and other Rickettsia bacterial antigen components. The bacterial antigens described herein also include any other bacterial, mycobacterial, mycoplasma, rickettsial, or chlamydial antigen.In other embodiments, the second target binds to bacteria selected from Escherichia coli, Pseudomonas bacteria, Staphylococcal bacteria, Enterobacteriaceae bacteria, Streptococcus bacteria, Haemophilus influenzae, Leptospira interrogans, Legionella bacteria, Mycobacterium tuberculosis, Candida albicans bacteria, Acinetobacter baumannii bacteria, Stenotrophomonas maltophilia bacteria, Clostridium difficile bacteria, Enterococcus bacteria, Klebsiella pneumoniae bacteria, Necrotizing fascitis bacteria, Corynebacterium bacteria, Helicobacter pylori bacteria, Campylobacter bacteria, Salmonellae bacteria, Neisseria gonorrhoeae bacteria, Haemophilus influenza bacteria, Shigella bacteria, and combinations thereof.

[0162] Exemplary fungal antigens that can be used as a second target include, but are not limited to, Candida fungal antigen components; Histoplasma fungal antigens such as heat shock protein 60 (HSP60) and other Histoplasma fungal antigen components; Cryptococcus fungal antigens such as capsular polysaccharide and other Cryptococcus fungal antigen components; Coccidioides fungal antigens such as spherule antigen and other Coccidioides fungal antigen components; and dermatophyte fungal antigens such as trichophytin and other Coccidioides fungal antigen components. Other fungal targets can be from fungal infections, which include Alternaria alternata, Aspergillus fumigatus, Aspergillus niger, Aspergillus flavus, Aspergillus nidulans, Aspergillus paraciticus, Candida albicans, Candida dubliniensis, Candida famata, Candida glabrata, Candida guilliermondii, Candida haemulonii, Candida kejyr, Candida krusei, Candida lusitaniae, Candida norvegensis, Candida parapsilosis, Candida tropicalis, Candida viswanathii, Epidermophyton floccosum, Fusarium graminearum, Fusarium oxysporum, Fusarium solani, Fusarium monoliforme, Trichophyton rubrum, Trichophyton mentagrophytes, Trichophyton inter digitales, Trichophyton tonsurans, Cryptococcus neoformans, Cryptococcus gattiigattii), Cryptococcus grubii, Colletotrichum graminicola, Microsporum canis, Microsporum gypseum, Penicillium marneffei, Trichosporon beigelii, Trichosporon asahii, Trichosporon inkin, Trichosporon asteroides, Trichosporon cutaneum, Trichosporon domesticum, Trichosporon mucoides, Trichosporon ovoides, Trichosporon pullulans, Trichosporon loubieri, Trichosporon japonicum, Scedosporium apiospermum, Scedosporium prolifwans, Paecilomyces variotii, Paecilomyces lilacinus, Acremonium stricutm, Cladophialophora bantiana, Wangiella dermatitidis, Ramichloridium obovoideum, Chaetomium atrobrunneum, Dactlaria gallopavum, Bipolaris spp, Exserohilum rostratum, Absidia corymbifera, Apophysomyces elegans, Mucor indicus, Rhizomucor pusillus, Rhizopus oryzae, Cunninghamellabertholletiae), Cokeromyces recurvatus, Saksenaea vasiformis, Syncephalastrum racemosum, Basidiobolus ranarum, Conidiobolus coronatusl, Conidiobolus incongruus, Blastomyces dermatitidis, Coccidioides immitis, Coccidioides posadasii, Histoplasma capsulatum, Paracoccidioides brasiliensis, Pseudallescheria boydii, Sporothrix schenckii, Alternaria brasicicola, Alternaria alternata, Aspergillus nidulans, Botrytis cinerea, Cercospora beticola, Cercospora zeae maydis, Cochliobolus heterostrophus, Exserohilum turcicum, Fusarium culmorum, Fusarium oxysporum, Fusarium oxysporum f.sp.dianthi, Fusarium solani, Fusarium pseudograminearum, Fusarium verticilloides, Gaeumannomyces graminis var.tritici, Plasmodiophora brassicae, Sclerotinia sclerotiorum, Stenocarpella (Diplodia) maydis, Thielaviopsisbasicola), Verticillium dahliae, Ustilago zeae, Puccinia sorghi, Macrophomina phaseolina, Phialophora gregata, Diaporthe phaseolorum, Cercospora sojina, Phytophthora sojae, Rhizoctonia solani, Phakopsora pachyrhizi, Alternaria macrospora, Cercospora gossypina, Phoma exigua, Puccinia schedonnardii, Puccinia cacabata, Phymatotrichopsis omnivora, Fusarium avenaceum, Alternaria brassicae, Alternaria raphani, Blumeria graminis, Septoria tritici, Septoria nodorum, Mycosphaerella zeae, Rhizoctonia cerealis, Ustilago tritici, Puccinia graminis, Puccinia triticina, Tilletia indica, Tilletia caries, Tilletia controversa, Alternaria solani, Alternaria brassicae, Alternaria brassicae, Monilinia fructicola, Venturia inaequalis, Cladosporum carpophilum, Botryosphaeria obtuse, Moniliniavaccinia - corymbosi), Sclerotinia homeocarpa, Podosphaera xanthii, Podosphaera fuliginea, Erysiphe cichoracearum, Blumeria graminis f.sp. Tritici, Blumeria graminis f.sp. Hordei, Microsphaera diffusa, Erysiphe necator, Leveillula taurica, Podosphaera leucotricha, Podosphaera aphanis, Sawadaea tulasnei, Erysiphe berberidis, Golovinomyces orontii, Peronospora belbahrii, Pseudoperonospora cubensis, Plasmopara viticola, Pseudoperonospora humuli, Peronospora manshurica, Pryonospora halstedii, Phytopthora capsici, Phytopthora infestans, Phytopthora cinnamomi, Phytopthora sojae, Phytopthora agathidicida, Phytopthora cactorum, Phytopthora citricola, Phytopthora fragariae, Phytopthora kernoviae, Phytopthora lateralis, Phytopthora megakarya, Phytopthoramultivora), Phytophthora nicotianae, Phytophthora palmivora, Phytophthora ramorum, Phytophthora quercina, and combinations thereof.

[0163] Exemplary parasite antigens that can be used as a second target include, but are not limited to, Plasmodium falciparum antigens such as merozoite surface antigen, sporozoite surface antigen, circumsporozoite antigen, gametocyte / gamete surface antigen, blood-stage antigen pf 155 / RESA, and other Plasmodium antigen components; Toxoplasma gondii antigens such as SAG-1, p30, and other Toxoplasma gondii antigen components; Schistosoma antigens such as glutathione-S-transferase, paramyosin, and other Schistosoma antigen components; Leishmania major and other Leishmania antigens such as gp63, lipophosphoglycan, and related proteins and other Leishmania antigen components; and Trypanosoma cruzi antigens such as 75-77 kDa antigen, 56 kDa antigen, and other Trypanosoma antigen components.

[0164] Exemplary tick antigens that can be used as a second target include, but are not limited to, antigens from ticks of the following: blood-sucking arthropods such as ticks selected from the group consisting of Ixodes, Bothriocrotoninae, Amblyomminae, Haemaphysalinae, Rhipicephalinae (including Hyalomminae), Nuttalliellidae, Argasinae, Otobinae, Antricolinae, Nothhoaspinae, and Ornithodorinae; blacklegged ticks from the following species: Ixodes, Dermacentor variabillis, Rhipicephalus sanguineus, or Amblyomma americanum; blacklegged ticks selected from the group consisting of Ixodes scapularis, Ixodes pacificus, Ixodes ricinus, and Ixodes persulcatus.

[0165] In some embodiments, the second target is a tumor cell antigen, which can be any molecule whose expression is restricted to or overexpressed in cancer cells. Many tumor-associated antigens have been identified and described in the literature, and thus are not all listed herein. In some embodiments, the tumor antigen is a tumor-specific antigen (TSA) or a tumor-associated antigen (TAA). A TSA is unique to tumor cells and does not occur on other cells in the body. A TAA-related antigen is not unique to tumor cells but is also expressed on normal cells under conditions that do not induce an immune tolerance state to the antigen. The expression of the antigen on the tumor can occur under conditions that enable the immune system to respond to the antigen. TAAs can be antigens that are expressed on normal cells during fetal development when the immune system is not yet mature enough to respond, or they may be antigens that are normally present at very low levels on normal cells but are expressed at much higher levels on tumor cells.

[0166] In some embodiments, the second target is selected from tumor-associated surface antigens such as 5T4, alpha-fetoprotein (AFP), B7-1 (CD80), B7-2 (CD86), BCMA, beta-human chorionic gonadotropin, CA-125, carcinoembryonic antigen (CEA), CD123, CD133, CD138, CD19, CD20, CD22, CD23, CD24, CD25, CD30, CD33, CD34, CD4, CD40, CD44, CD56, CD8, CLL-1, c-Met, CMV-specific antigen, CS-1, CSPG4, CTLA-4, DLL3, disialoganglioside GD2, ductal epithelial mucin, EBV-specific antigen, EGFR variant III (EGFRvIII), ELF2M, endothelial factor, ephrin B2, epidermal growth factor receptor (EGFR), epithelial cell adhesion molecule (EpCAM), epithelial tumor antigen, ErbB2 (HER2 / neu), fibroblast activation protein (fap), FLT3, folate-binding protein, GD2, GD3, glioma-associated antigen, glycosphingolipid, gp36, HBV-specific antigen, HCV-specific antigen, HER1-HER2, HER2-HER3 combination, HERV-K, high molecular weight melanoma-associated antigen (HMW-MAA), HIV-1 envelope glycoprotein gp41, HPV-specific antigen, human telomerase reverse transcriptase, IGFI receptor, IGF-II, IL-11Rα, IL-13R-a2, influenza virus-specific antigen;CD38, insulin-like growth factor (IGF1)-I, intestinal carboxylesterase, kappa chain, LAGA-la, lambda chain, Lassa virus-specific antigen, lectin-reactive AFP, lineage-specific or tissue-specific antigens (such as CD3), MAGE, MAGE-A1, major histocompatibility complex (MHC) molecules, MHC molecules presenting tumor-specific peptide epitopes, M-CSF, melanoma-associated antigens, mesothelin, MN-CAIX, MUC-1, mut hsp70-2, mutant p53, mutant ras, neutrophil elastase, NKG2D, Nkp30, NY-ESO-1, p53, PAP, prostate enzymes, prostate-specific antigen (PSA), prostate cancer tumor antigen-1 (PCTA-1), prostate-specific antigen protein, STEAP1, STEAP2, PSMA, RAGE-1, ROR1, RU1, RU2(AS), surface adhesion molecules, survivin and telomerase, TAG-72, extra domain A (EDA) and extra domain B (EDB) of fibronectin, and A1 domain of tenascin-C (TnC A1), thyroglobulin, tumor stroma antigen, vascular endothelial growth factor receptor-2 (VEGFR2), virus-specific surface antigens such as HIV-specific antigens (e.g., HIV gp120), and any derivatives or variants of these surface antigens; differentiation antigens such as MART-1 / MelanA (MART-I), gp100 (Pmel 17), tyrosinase, TRP-1, TRP-2, and tumor-specific multi-lineage antigens such as MAGE-1, MAGE-3, BAGE, GAGE-1, GAGE-2, p15; overexpressed embryonic antigens such as CEA; overexpressed oncogenes and mutant tumor suppressor genes such as p53, Ras, HER2 / neu; unique tumor antigens resulting from chromosomal translocations such as BCR-ABL, E2A-PRL, H4-RET, IGH-IGK, MYL-RAR; and virus antigens such as Epstein-Barr virus antigen EBVA and human papillomavirus (HPV) antigens E6 and E7;TSP-180, MAGE-4, MAGE-5, MAGE-6, RAGE, NY-ESO, pl85erbB2, pl80erbB-3, c-met, nm-23HI, PSA, TAG-72, CA 19-9, CA 72-4, CAM 17.1, NuMa, K-ras, beta-Catenin, CDK4, Mum-1, p15, p16, 43-9F, 5T4, 791Tgp72, alpha-fetoprotein, beta-HCG, BCA225, BTAA, CA125, CA 15-3\CA 27.29\BCAA, CA 195, CA242, CA-50, CAM43, CD68\P1, CO-029, FGF-5, G250, Ga733\EpCAM, HTgp-175, M344, MA-50, MG7-Ag, MOV18, NB / 70K, NY-CO-1, RCAS1, SDCCAG16, TA-90\Mac-2 binding protein\Cysteine protease C-related protein, TAAL6, TAG72, TLP, TPS; GPC2, CD276, delta-like protein ligand 3 (DLL3), NY-ESO-1, melanoma-associated antigen 4; survivin protein, synovial sarcoma X breakpoint protein 2, CD3, epidermal growth factor receptor (EGFR), erbb2 tyrosine kinase receptor, HER2, CEA, CD66, CD66e, ROR1, ntrkr1 tyrosine kinase receptor, GPC3, mesothelin, glutamate carboxypeptidase II, PMSA, PD-L1, folate receptor alpha, PSCA, mucin 1, HLA antigens (such as HLA class I antigen A-2 alpha, HLA class I antigen A-11 alpha, HLA class II antigens), c-Met, hepatocyte growth factor receptor, K-Ras GTPase (KRAS), IL-15 receptor, Kit tyrosine kinase, PDGF receptor beta, RET tyrosine kinase receptor; Raf 1 protein kinase, Raf B protein kinase, thymidylate synthase, topoisomerase II, Brachyury protein, Flt3 tyrosine kinase, VEGF, VEGF receptors (VEGF-1 receptor, VEGF-2 receptor and VEGF-3 receptor), estrogen receptor, neoantigen, human papillomavirus E6 and heat shock protein.;

[0167] Antibody-dependent cell phagocytosis in the immune response triggered by antibody repurposing

[0168] In some embodiments, repurposed antibodies for passive infusion can be used in antitoxin, antiviral, anti - cancer, and anti - inflammatory therapies. Without being bound by theory, repurposed antibodies can exert their functions through multiple mechanisms. In some embodiments, repurposed antibodies act through target neutralization, which mainly depends on the interaction between a second target - binding molecule (such as Technein) and the second target, and thus is mainly independent of the Fc domain. In other embodiments, including antibody - dependent cell - mediated cytotoxicity (ADCC) and antibody - dependent cell phagocytosis (ADCP), the Fc domain of the repurposed antibody needs to be recognized through Fc receptors to interact with other proteins or immune effector cells. These Fc - receptor - dependent antibody functions provide a direct link between the innate immune system and the adaptive immune system, leveraging the powerful anti - pathogen functions of the innate immune system and overcoming its inherently limited pattern - recognition capabilities by harnessing the diversity and specificity of the adaptive immune response. Fc - receptor - dependent antibody functions are an important part of the immune response and can provide mechanisms for clearing infected host cells, immune complexes, or opsonized pathogens. Fc - receptor - dependent antibody functions also participate in activating downstream adaptive immune responses by promoting antigen presentation or stimulating the secretion of inflammatory mediators.

[0169] In one embodiment, the Fc - receptor - dependent function of antibody - dependent cell phagocytosis (ADCP) provides a mechanism for using repurposed antibodies to clear pathogens (such as viruses) and pathogen - infected cells, as well as a mechanism for stimulating downstream adaptive immune responses by promoting antigen presentation or stimulating the secretion of inflammatory mediators. In some embodiments, ADCP mediated by repurposed antibodies may be a mechanism for inhibiting pathogen infections (such as viral infections).

[0170] In some embodiments, the antibody is of the IgG1 isotype. In some embodiments, the antibody is of the IgG2 isotype. In some embodiments, the antibody is of the IgG3 isotype. In some embodiments, the antibody is of the IgG4 isotype. In some embodiments, the antibody is of the IgA1 isotype. In some embodiments, the antibody is of the IgA2 isotype. For human IgG isotypes, IgG3 has the highest affinity for most type I FcγRs, followed by IgG1, then IgG4, and then IgG2. In contrast, IgA subclasses are not the main factors regulating ADCP because human FcαR has been shown to have similar affinities for IgA1 and IgA2. In some embodiments, the antibody is of the IgA or IgM isotype.

[0171] In some embodiments, the antibody is glycosylated. For IgG antibodies, there are 36 possible glycosylation forms and 4 different subclasses, resulting in a total of 144 possible unique Fc regions. Any of these Fc regions can be used in the antibodies of the present disclosure.

[0172] In some embodiments, Technein is specifically selected for its significant binding to an epitope that is particularly effective in engaging a repurposed antibody to induce ADCP. In one embodiment, for ADCP of virions, the epitope may be present on the surface of the virion, while for ADCP of infected cells, any epitope expressed on the surface of the virus-infected cells may be a potential target. In one embodiment, Technein is selected for its activity in supporting ADCP of pathogen (e.g., virus) particles. In one embodiment, Technein is selected for its activity in supporting ADCP of pathogen-infected cells.

[0173] In some embodiments, it is known that the original antibody mediates ADCP. In some embodiments, the antibody is trastuzumab, rituximab, cetuximab, or ipilimumab.

[0174] Methods for assessing ADCP

[0175] In some embodiments, the ability of the repurposed antibody to mediate ADCP is determined by the method described in the examples. In other embodiments, the method is as described in Guillaume Beaudoin-Bussières, Jonathan Richard, Jérémie Prévost, Guillaume Goyette, Andrés Finzi, A new flow cytometry assay to measure antibody-dependent cellular cytotoxicity against SARS-CoV-2 Spike-expressing cells, STAR Protocols, Volume 2, Issue 4, 2021, 100851. In other embodiments, the method is as described in Duchemin M, Tudor D, Cottignies-Calamarte A, Bomsel M. Antibody-Dependent Cellular Phagocytosis of HIV-1-Infected Cells Is Efficiently Triggered by IgA Targeting HIV-1 Envelope Subunit gp41. Front Immunol. June 9, 2020; 11:1141. doi:10.3389 / fimmu.2020.01141. PMID: 32582208; PMCID: PMC7296124. These references are incorporated herein by reference in their entirety.

[0176] Antibody-dependent cell-mediated cytotoxicity (ADCC)

[0177] Antibody-dependent cell-mediated cytotoxicity (ADCC), also known as antibody-dependent cell-mediated cytotoxicity, is an immune mechanism by which effector cells bearing Fc receptors can recognize and kill antibody-coated target cells expressing tumor or pathogen-derived antigens on their surface. In one embodiment, an antibody is selected for repurposing because of its ability to trigger ADCC against tumor cells. In other embodiments, an antibody is selected for repurposing because of its ability to trigger ADCC against pathogen-derived antigens on the surface of infected cells. In one embodiment, the original antibody does not have the ability to mediate ADCC, but the repurposed antibody can mediate ADCC. In some embodiments, both antibodies can mediate ADCC.

[0178] In some embodiments, the original antibody has been shown to enhance ADCC. Examples of FDA-approved antibodies known to mediate ADCC include trastuzumab (anti-HER2), rituximab (anti-CD20), cetuximab (anti-EGFR), avelumab (anti-PD-L1), ofatumumab (anti-CD20), obinutuzumab (anti-CD20), mogamulizumab (anti-CCR4), margetuximab (anti-HER2), ublituximab (anti-CD20).

[0179] Measuring ADCC

[0180] In some embodiments, the ability of the repurposed antibody to mediate ADCC is determined by the methods described in the examples. In some other embodiments, ADCC is measured by other methods selected from labeling target cells with a fluorescent dye, which allows for the sensitive determination of cytotoxicity by flow cytometry ( K, Garritsen, H, Van Graft, M, et al. A simple and sensitive flow cytometric assay for the determination of the cytotoxic activity of human natural killer cells. J Immunol Methods 1968;135:81–9.); The VITAL assay, which was developed to measure cytotoxicity of multiple target populations simultaneously both in vitro and in vivo (Hermans, I, Silk, J, Yang, J, et al. The VITAL assay: a versatile fluorometric technique for assessing CTL- and NKT-mediated cytotoxicity against multiple targets in vitro and in vivo. J Immunol Methods 2004;285:25–40); The 51Cr release assay but replace radioisotopes with natural cell products; Combining flow cytometric and labeling-based techniques for high-throughput single-cell computer image analysis (Welter, A, Sundararaman, S, Li, R, et al.High-throughput GLP-capable target cell visualization assay for measuring cell-mediated cytotoxicity (Cells 2018; 7:35); genetically modified target cells to express a reporter protein for a standardized assay (Rossignol, A, Bonnaudet, V, Clémenceau, B et al. A high-performance, non-radioactive potency assay for measuring cytotoxicity: a full substitute of the chromium-release assay targeting the regulatory-compliance objective. MAbs 2017; 9:521–35). Alternatively, when cells are killed, they release lactate dehydrogenase and other proteases, which can be quantified by providing a fluorescent substrate, thus more accurately assessing cytotoxicity without any labeling or manipulation of the target cells (Hassenrück, F,. E, E, et al. Sensitive detection of the natural killer cell-mediated cytotoxicity of anti-CD20 antibodies and its impairment by B-cell receptor pathway inhibitors. BioMed Res Int 2018; 2018: 1–9; Niles, A, Moravec, R, Eric Hesselberth, P, et al. A homogeneous assay to measure live and dead cells in the same sample by detecting different protease markers. Anal Biochem 2007; 366: 197–206).

[0181] Targeted blocking and / or neutralization

[0182] In some embodiments, the repurposed antibody is a blocking antibody. In some embodiments, the repurposed antibody is a neutralizing antibody. In some embodiments, a blocking antibody does not trigger a response when binding its target, but prevents other molecules (e.g., another antibody) from interfering with the target. In some embodiments, a blocking antibody binds to its target and directly interferes with its function, such as blocking cell adhesion or receptor-ligand binding. In some embodiments, a neutralizing antibody binds to its target and abrogates its downstream cellular effects, such as cell proliferation or chemotaxis. In some embodiments, a neutralizing antibody is an endogenous antibody that is naturally produced by the body as part of the body's immune response to a drug or pathogen. In some embodiments, the antibody (original and / or repurposed antibody) is a neutralizing and / or blocking antibody.

[0183] In some embodiments, the original and / or repurposed antibody is a blocking antibody. Non-limiting examples of blocking antibodies include anti-PD-L, anti-PD-L1, and anti-CTLA4 antibodies, including nivolumab, pembrolizumab, ipilimumab, tremelimumab. In some embodiments, the blocking antibody is a complement-blocking antibody. In some embodiments, the antibody is capable of binding to and sequestering plasma proteins or plasma drugs. In one embodiment, the antibody is selected from adalimumab, pegylated certolizumab, golimumab, infliximab that bind tumor necrosis factor or bevacizumab that binds VEGF.

[0184] In one embodiment, the present disclosure provides neutralizing antibodies that can be used for virus clearance and achieve protection against multiple viruses. They can achieve this in a variety of ways, including interfering with virion binding to receptors, blocking virus uptake by host cells, and preventing viral genome uncoating in endosomes or causing viral particle aggregation. In one embodiment, the neutralizing antibody blocks the interaction of the target with its natural receptor. In some embodiments, the neutralizing antibody binds to the virus in a manner that blocks virus infection. In some embodiments, the neutralizing antibody binds to the viral capsid in a manner that inhibits viral genome uncoating. In some embodiments, the neutralizing antibody binds to the virus to produce virus complexes that can be destroyed by phagocytes.

[0185] Accordingly, in some embodiments, the immunotherapeutic agents / compounds or compositions of the present disclosure (i.e., synthetic compounds, repurposed antibodies, and their compositions) can act in cancer by triggering an immune response to disrupt or reduce the incidence of cancer cell growth. In some embodiments, the immunotherapeutic agents of the present disclosure can be used for any of the following applications:

[0186] Methods of treating or reducing the incidence of cancer using the immunotherapeutic compounds or compositions of the present disclosure.

[0187] In some embodiments, these claims cover methods of ameliorating, treating, or reducing the incidence of malignancy in a human subject, wherein the steps of the method assist or enhance the immune system to eradicate cancer cells. Examples include:

[0188] (a) A method of ameliorating, treating, or reducing the incidence of cancer / tumor / malignancy in a human subject, wherein the steps of the method assist or enhance the immune system to eradicate cancer cells, and wherein the method comprises administering a therapeutically effective amount of an antibody, synthetic compound, cell, and / or nucleic acid as described in the present disclosure, which triggers an active (or achieves passive) immune response to destroy cancer cells;

[0189] b) A method of co-administering a biological adjuvant (such as interleukin, cytokine, Bacillus Calmette-Guérin, monophosphoryl lipid A, etc.) in combination with conventional cancer therapies (such as chemotherapy, radiotherapy, or surgery).

[0190] c) A method involving administering any vaccine, wherein the vaccine disrupts cancer cell growth or reduces the incidence of cancer cell growth by activating the immune system, and wherein the vaccine comprises a therapeutically effective amount of an antibody, synthetic compound, cell, and / or nucleic acid as described in the present disclosure; and

[0191] (d) A method of treating cancer by using a therapeutically effective amount of the antibodies, synthetic compounds, cells, and / or nucleic acids described in the present disclosure as adoptive immunotherapy in vivo or ex vivo, which includes using autologous and / or allogeneic cells or immortalized cell lines in a subject in need thereof, which includes administering or giving to the subject the antibodies, synthetic compounds, cells, and / or nucleic acids described in the present disclosure and / or contacting immunotherapeutic cells with the synthetic compounds described in the present disclosure.

[0192] (ii) The immunotherapeutic compounds or compositions of the present disclosure for cancer immunotherapy, wherein the compounds or compositions comprise the repurposed antibodies, fragments thereof, and / or synthetic compounds of the present disclosure, or CAR-T, T cells, or TILs that are contacted or conjugated with the compounds of the present disclosure.

[0193] Indications

[0194] As described above, the repurposed antibodies can be used for a variety of indications. In some embodiments, the indications are selected from infectious diseases, cancer, hematological malignancies, autoimmune diseases, hypercholesterolemia, asthma, osteoporosis, inflammatory bowel disease, allograft rejection, and drug reversal. In some embodiments, although the targets are different, the repurposed antibodies and the original antibodies can be used for the same indication. In some embodiments, the repurposed antibodies have acquired the ability to bind to those antibodies selected from Tables 1 to 3 and have the same function because it has been repurposed to bind to the same target, even if it is completely different from the original indication.

[0195] In some embodiments, the repurposed antibodies are prepared in vitro by binding the original antibodies to the synthetic compounds described in the present disclosure in vitro. In some embodiments, the repurposed antibodies are prepared in vivo by administering the original antibodies before, after, and / or simultaneously with the synthetic compounds described in the present disclosure.

[0196] The repurposed antibodies and / or their components (such as antibodies, first molecules, second molecules, and synthetic compounds) can be administered by any route. In some embodiments, the repurposed antibodies and / or their components are administered by the same route. In some embodiments, the repurposed antibodies and / or their components are administered orally, intravenously, subcutaneously, and / or intramuscularly.

[0197] In one embodiment, the repurposed antibodies of the present disclosure can be used to treat infections (e.g., viruses, bacteria, fungi, etc.). In other embodiments, the repurposed antibodies of the present disclosure can be used to treat one or more of the following diseases or conditions: amyotrophic lateral sclerosis; endotoxicosis; atherosclerotic vascular disease or coronary artery, cancer (see elsewhere); rheumatoid arthritis; arterial disease; in-stent restenosis; carotid metabolic disease; stroke; acute myocardial infarction; heart failure; peripheral arterial disease; limb ischemia; vein graft failure; AV fistula failure; Crohn's disease; ulcerative colitis; ileitis and enteritis; vaginitis; psoriasis and inflammatory skin diseases such as dermatitis; eczema; atopic dermatitis; allergic contact dermatitis; urticaria; vasculitis; spondyloarthropathy; scleroderma; respiratory allergic diseases such as asthma; allergic rhinitis; allergic lung disease; arthritis (e.g., rheumatoid arthritis and psoriatic arthritis); eczema; psoriasis; osteoarthritis; multiple sclerosis; systemic lupus erythematosus; diabetes; glomerulonephritis; transplant rejection (including allograft rejection and graft-versus-host disease) or engineered tissue rejection; infectious diseases; myositis; inflammatory central nervous system diseases; stroke; closed head injury; neurodegenerative diseases; Alzheimer's disease; encephalitis; meningitis; osteoporosis; gout; hepatitis; hepatic veno-occlusive disease (VOD); hemorrhagic cystitis; nephritis; sepsis; sarcoidosis; conjunctivitis; otitis; chronic obstructive pulmonary disease; sinusitis; Bechet's syndrome; graft-versus-tumor effect; mucositis; appendicitis; appendiceal rupture; peritonitis; aortic valve disease; mitral valve disease; Rett syndrome; tuberous sclerosis; phenylketonuria; Smith-Lemli-Opitz syndrome and fragile X chromosome syndrome; Parkinson's disease; Aicardi-Goutières Syndrome; Alexander Disease; Allan-Hemdon-Dudley Syndrome; POLG-related disorders; alpha-mannosidosis (types II and III); Alström syndrome Angelman Syndrome; Ataxia-Telangiectasia; Neuronal Ceroid-Lipofuscinosis; β-Thalassemia; Bilateral Optic Atrophy and (infantile) Optic Atrophy Type 1; Retinoblastoma (bilateral); Canavan Disease; Cerebro-Oculo-Facioskeletal Syndrome 1 [COFS1]; Cerebrotendinous Xanthomatosis; Cornelia de Lange Syndrome; MAPT-Related Disorders; Hereditary Prion Diseases; Dravet Syndrome; Early-Onset Familial Alzheimer's Disease; Friedreich Ataxia [FRDA]; Frynns Syndrome; Fucosidosis; Fukuyama Congenital Muscular Dystrophy; Galactosialidosis; Gaucher Disease; Organic Acidemias; Hemophagocytic Lymphohistiocytosis; Hutchinson-Gilford Progeria Syndrome; Mucolipidosis II; Infantile Free Sialic Acid Storage Disease; PLA2G6-Related Neurodegeneration; Jervell and Lange-Nielsen Syndrome; Junctional Epidermolysis Bullosa; Huntington Disease; Krabbe Disease (infantile); Mitochondrial DNA-Related Leigh Syndrome and NARP; Lesch-Nyhan Syndrome; LIS1-Related Lissencephaly; Lowe Syndrome; Maple Syrup Urine Disease; MECP2 Duplication Syndrome; ATP7A-Related Copper Transport Disorder; LAMA2-Related Muscular Dystrophy; Arylsulfatase A Deficiency; Mucopolysaccharidosis Type I, II or III; Peroxisome Biogenesis Disorders; Zellweger Syndrome Spectrum; Neurodegeneration with Brain Iron Accumulation Disorders; Acid Sphingomyelinase Deficiency; Niemann-Pick Disease Type C; Glycine Encephalopathy; ARX-Related Disorders; Urea Cycle Disorders; COL1A1 / 2-Related Osteogenesis Imperfecta; Mitochondrial DNA Deletion Syndromes; PLP1-Related Disorders; Perry Syndrome; Phelan-McDermid Syndrome;Glycogen storage disease type II (Pompe disease) (infantile); MAPT-related disorders; MECP2-related disorders; Acrofacial dysostosis type 1; Roberts syndrome; Sandhoff disease; Schindler Disease-Type 1; Adenosine deaminase deficiency; Smith-Lemli-Opitz Syndrome; Spinal muscular atrophy; Infantile spinocerebellar ataxia; Hexosaminidase A deficiency; Thanatophoric dysplasia type 1; Type VI collagen-related disorders; Usher Syndrome Type I; Congenital muscular dystrophy; Wolf-Hirschhorn Syndrome; Lysosomal Acid Lipase Deficiency; and Xeroderma Pigmentosum.;

[0198] Pharmaceutical Compositions and Formulations

[0199] Compositions are provided herein that comprise an original antibody and / or a synthetic molecule (or portion thereof) of the present disclosure, a repurposed antibody or a target-binding fragment thereof as described herein, and another component such as a carrier. Pharmaceutical compositions / therapeutic formulations are also provided that comprise an original antibody and / or a synthetic molecule (or portion thereof) of the present disclosure, a repurposed antibody or a target-binding fragment thereof as described herein, and an excipient and / or a diluent. In one embodiment, the carrier is not a naturally occurring compound. In one embodiment, the excipient is not a naturally occurring compound. In another embodiment, the diluent is not a naturally occurring compound. In another embodiment, a formulation comprising a repurposed antibody or a target-binding fragment thereof as described herein does not contain naturally occurring compounds other than optionally water. It will be apparent to those skilled in the art that certain carriers, excipients or diluents may be more preferred, depending, for example, on the route of administration and the concentration of the antibody administered.

[0200] In one embodiment, a therapeutic formulation of the native antibody and / or synthetic molecule (or portion thereof) of the present disclosure, repurposed antibody or target-binding fragment thereof is prepared for storage and / or administration in the form of a lyophilized formulation or an aqueous solution by mixing the native antibody and / or synthetic molecule (or portion thereof) of the present disclosure, repurposed antibody or target-binding fragment thereof having the desired purity with an optional pharmaceutically acceptable carrier, diluent, excipient or stabilizer (Remington's Pharmaceutical Sciences, 16th Edition, Osol, A. (ed.) (1980)). In one embodiment, at the dosages and concentrations employed, the acceptable carriers, excipients or stabilizers are non-toxic to the recipient and include: buffers such as phosphate, citrate and other organic acid buffers; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyl dimethyl benzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl alcohol or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); polypeptides of low molecular weight (less than about 10 residues); proteins such as serum albumin, gelatin or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine or lysine; monosaccharides, disaccharides and other sugars including glucose, mannose or dextrin; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol; salt-forming counterions such as sodium; metal complexes (e.g., Zn-protein complexes); and / or nonionic surfactants such as TWEEN, PLURONICS or polyethylene glycol (PEG). Examples of lyophilized antibody formulations are described in WO 97 / 04801, which is hereby incorporated by reference in its entirety.

[0201] In other embodiments, the formulation further comprises a surfactant. The surfactant can be selected, for example, from detergents, ethoxylated castor oil, polyethylene glycol glycerol esters, acetylated monoglycerides, sorbitan fatty acid esters, polyoxypropylene-polyoxyethylene block polymers (e.g., poloxamers such as F68, poloxamers 188 and 407, Triton X-100), polyoxyethylene sorbitan fatty acid esters, polyoxyethylene and polyethylene derivatives such as alkylated and alkoxylated derivatives (Tween, e.g., Tween-20, Tween-40, Tween-80 and Brij-35), monoglycerides or their ethoxylated derivatives, diglycerides or their polyoxyethylene derivatives, alcohols, glycerol, lectins and phospholipids (e.g., phosphatidylserine, phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, diphosphatidylglycerol and sphingomyelin), phospholipid derivatives (e.g., dipalmitoyl phosphatidic acid) and lysophospholipids (e.g., palmitoyl lysophosphatidyl-L-serine and 1-acyl-sn-glycero-3-phosphates of ethanolamine, choline, serine or threonine) and alkyl, alkoxy (alkyl ester), alkoxy (alkyl ether) derivatives of lysophosphatidyl and phosphatidylcholine, e.g., lauroyl and myristoyl derivatives of lysophosphatidylcholine, dipalmitoyl phosphatidylcholine, and modifications of the polar head group, i.e., choline, ethanolamine, phosphatidic acid, serine, threonine, glycerol, inositol and positively charged DODAC, DOTMA, DCP, BISHOP, lysophosphatidylserine and lysophosphatidylthreonine and glycerophospholipids (e.g., cephalin), glyceroglycolipids (e.g., galactosides), glycosphingolipids (e.g., ceramides, gangliosides), dodecylphosphocholine, egg lysolecithin, fusidic acid derivatives—(e.g., sodium taurodihydrofusidate, etc.), C6-C12 long-chain fatty acids and their salts (e.g., oleic acid and octanoic acid), acylcarnitine and its derivatives, Nα-acylated derivatives of lysine, arginine or histidine, or side-chain acylated derivatives of lysine or arginine, Nα-acylated derivatives of dipeptides containing any combination of lysine, arginine or histidine with neutral or acidic amino acids, Nα-acylated derivatives of tripeptides containing any combination of neutral amino acids with two charged amino acids, DSS (sodium docusate, CAS registry number [577-11-7]), calcium docusate, CAS registry number [128-49-4]), potassium docusate, CAS registry number [7491-09-0]), SDS (sodium dodecyl sulfate or sodium lauryl sulfate), sodium octanoate, cholic acid or its derivatives, bile acids and their salts and glycine or taurine conjugates, ursodeoxycholic acid, sodium cholate, sodium deoxycholate, sodium taurocholate, sodium glycocholate, N-hexadecyl-N,N-dimethyl-3-ammonio-1-propanesulfonate, anionic (alkylaryl sulfonate) monovalent surfactants, zwitterionic surfactants (e.g., N-alkyl-N,N-dimethylammonio-1-propanesulfonate, 3-cholamidopropyl dimethylammonio-1-propanesulfonate, cationic surfactants (quaternary ammonium bases) (e.g., cetyltrimethylammonium bromide, cetylpyridinium chloride), nonionic surfactants (e.g., dodecyl.β.-D-glucopyranoside), poloxamines (e.g., Tetronic's), which are tetrafunctional block copolymers obtained by the sequential addition of propylene oxide and ethylene oxide to ethylenediamine, or the surfactant can be selected from the group of imidazoline derivatives, or mixtures thereof. In one embodiment, the surfactant is not a naturally occurring compound. These specific surfactants each constitute an alternative embodiment of the present disclosure.

[0202] One embodiment provides a stable formulation of an original antibody and / or a synthetic molecule (or a portion thereof) of the present disclosure, a repurposed antibody or a target-binding fragment thereof, preferably comprising a phosphate buffer containing saline or a selected salt, and a preservation solution and formulation containing a preservative, and a multi-purpose preservation formulation suitable for pharmaceutical or veterinary use, which comprises at least one antibody and / or a target-binding fragment thereof in a pharmaceutically acceptable formulation. In one embodiment, the preservation formulation contains at least one known preservative in an aqueous diluent or optionally at least one preservative selected from the group consisting of phenol, m-cresol, p-cresol, o-cresol, chlorocresol, benzyl alcohol, phenylmercuric nitrate, phenoxyethanol, formaldehyde, chlorobutanol, magnesium chloride (such as hexahydrate), alkyl esters of hydroxybenzoic acid (methyl ester, ethyl ester, propyl ester, butyl ester, etc.), benzalkonium chloride, benzethonium chloride, sodium dehydroacetate, and thimerosal, or a mixture thereof. Any suitable concentration or mixture known in the art can be employed, such as 0.001 - 5%, or any range or value therebetween, such as but not limited to 0.001, 0.003, 0.005, 0.009, 0.01, 0.02, 0.03, 0.05, 0.09, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.3, 4.5, 4.6, 4.7, 4.8, 4.9, or any range or value therebetween. Non-limiting examples include, no preservative, 0.1 - 2% m-cresol (e.g., 0.2, 0.3, 0.4, 0.5, 0.9, 1.0%), 0.1 - 3% benzyl alcohol (e.g., 0.5, 0.9, 1.1, 1.5, 1.9, 2.0, 2.5%), 0.001 - 0.5% thimerosal (e.g., 0.005, 0.01), 0.001 - 2.0% phenol (e.g., 0.05, 0.25, 0.28, 0.5, 0.9, 1.0%), 0.0005 - 1.0% alkyl esters of hydroxybenzoic acid (e.g., 0.00075, 0.0009, 0.001, 0.002, 0.005, 0.0075, 0.009, 0.01, 0.02, 0.05, 0.075, 0.09, 0.1, 0.2, 0.3, 0.5, 0.75, 0.9, 1.0%), etc. In one embodiment, one or more of the preservatives are not naturally occurring compounds.

[0203] In one embodiment, the original antibodies and / or synthetic molecules (or portions thereof) of the present disclosure, the repurposed antibodies of the present disclosure or target-binding fragments thereof can be incorporated into a pharmaceutical composition suitable for administration to a subject. In a common embodiment, the pharmaceutical composition comprises the original antibodies and / or synthetic molecules (or portions thereof) of the present disclosure, the repurposed antibodies of the present disclosure or target-binding fragments thereof, and a pharmaceutically acceptable carrier. In one embodiment, "pharmaceutically acceptable carrier" includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, etc. that are physiologically compatible. Additional examples of pharmaceutically acceptable carriers include one or more of water, saline, phosphate buffered saline, dextrose, glycerol, ethanol, etc. and combinations thereof. In many cases, an isotonic agent, such as sugar, a polyol such as mannitol, sorbitol, or sodium chloride, may preferably be included in the composition. The pharmaceutically acceptable carrier may also contain small amounts of auxiliary substances, such as wetting agents or emulsifying agents, preservatives or buffering agents, which may extend the shelf life or effectiveness of the antibody or its target-binding fragment.

[0204] In one embodiment, an appropriate amount of a pharmaceutically acceptable salt is used in the formulation to render the formulation isotonic. Examples of carriers include saline, Ringer's solution, and dextrose solution. In one embodiment, the pH of the solution is from about 5 to about 8. In another embodiment, the pH is from about 7 to about 7.5. Other carriers include sustained-release formulations, such as a semipermeable matrix of a solid hydrophobic polymer containing the antibody or its target-binding fragment, which matrix is in the form of a shaped article, such as a film, liposome, or microparticle. As used herein, a sustained-release matrix is a matrix made of a material (usually a polymer that degrades by enzymatic or acid / base hydrolysis or dissolution). Once inserted into the body, enzymes and body fluids act on the matrix. The sustained-release matrix is preferably selected from biocompatible materials, such as liposomes, polylactic acid (polylactide), polyglycolic acid (a polymer of glycolic acid), polylactic-co-glycolic acid (a copolymer of lactic acid and glycolic acid), polyanhydrides, poly(ortho)esters, polypeptides, hyaluronic acid, collagen, chondroitin sulfate, carboxylic acids, fatty acids, phospholipids, polysaccharides, nucleic acids, polyamino acids, amino acids such as phenylalanine, tyrosine, isoleucine, polynucleotides, polyvinyl propylene, polyvinyl pyrrolidone, and silicone. Preferred biodegradable matrices are matrices of one of polylactic acid, polyglycolic acid, or polylactic-co-glycolic acid (a copolymer of lactic acid and glycolic acid).

[0205] The compositions of the present disclosure can be in a variety of forms. This includes, for example, liquid, semi-solid, and solid dosage forms such as liquid solutions (e.g., injectable and infusible solutions), dispersions or suspensions, tablets, pills, powders, liposomes, and suppositories. In some embodiments, such compositions may also include buffers (e.g., neutral buffered saline or phosphate buffered saline), carbohydrates (e.g., glucose, mannose, sucrose, or dextran), mannitol, proteins, polypeptides, or amino acids (such as glycine), antioxidants, chelating agents (such as EDTA or glutathione), adjuvants (e.g., aluminum hydroxide), and / or preservatives. Alternatively, the compositions of the present disclosure can be formulated as lyophilized products. The native antibodies and / or synthetic molecules (or portions thereof), repurposed antibodies, or their target-binding fragments of the present disclosure can also be encapsulated within liposomes using well-known techniques.

[0206] Dosage forms suitable for oral administration generally contain from about 0.1 milligram to about 500 milligrams of the antibody or its target-binding fragment (active ingredient) per unit or container. In these pharmaceutical compositions, the active ingredient is generally present in an amount of about 0.5 - 99.999% by weight based on the total weight of the composition.

[0207] Under the conditions of manufacture and storage, the therapeutic compositions / formulations must generally be sterile and stable. The compositions can be formulated as solutions, microemulsions, dispersions, liposomes, or other ordered structures suitable for high drug concentrations. The required amount of the active compound (i.e., the antibody or its target-binding fragment) in a suitable solvent can be admixed with one or a combination of the above ingredients as needed, and then filtered sterilized to prepare a sterile injectable solution. Generally, a dispersion can be prepared by adding the active agent to a sterile vehicle containing a basic dispersion medium and the other required ingredients described above. In the preparation of sterile powders for sterile injectable solutions, the preferred methods of preparation are vacuum drying and freeze drying to obtain a powder of the active ingredient and any other required ingredients from a previously sterile filtered solution. The proper fluidity of the solution can be maintained, for example, by using a coating such as lecithin, by maintaining the required particle size in the case of a dispersion, and by using surfactants. Prolonged absorption of injectable compositions can be achieved by including substances that delay absorption, such as monostearates and gelatin.

[0208] The preferred dosage form depends on the intended mode of administration and therapeutic application. Those of ordinary skill in the art are familiar with the procedures for determining such dosages. Typical compositions are in the form of injectable or infusible solutions, such as those similar to those used for passive immunization of humans with other antibodies. The most typical mode of administration is parenterally (e.g., intravenously, subcutaneously, intraperitoneally, intramuscularly). In a preferred embodiment, the antibody is administered by intravenous infusion or injection. In another preferred embodiment, the antibody is administered by intramuscular or subcutaneous injection.

[0209] The original antibodies and / or synthetic molecules (or portions thereof) of the present disclosure, the repurposed antibodies of the present disclosure or their target-binding fragments can be administered by a variety of methods known in the art. Although for many therapeutic applications, the preferred route / mode of administration is intravenous injection or infusion. Those skilled in the art will appreciate that the route and / or mode of administration will vary depending on the desired outcome. In certain embodiments, the original antibodies and / or synthetic molecules (or portions thereof) of the present disclosure, the repurposed antibodies or their target-binding fragments can be formulated with a carrier that will protect the compound from rapid release, such as a controlled-release formulation, including implants, transdermal patches and microencapsulation delivery systems. Biodegradable biocompatible polymers such as ethylene-vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters and polylactic acid can be used. Many methods of preparing such formulations are patented or are well known to those skilled in the art. See, for example, Sustained and Controlled Release Drug Delivery Systems, edited by J.R. Robinson, Marcel Dekker, Inc., New York, 1978.

[0210] In certain embodiments, the original antibodies and / or synthetic molecules (or portions thereof) of the present disclosure, the repurposed antibodies of the present disclosure or their target-binding fragments can be administered orally, for example, in conjunction with an inert diluent or an absorbable edible carrier. The original antibodies and / or synthetic molecules (or portions thereof) of the present disclosure, the repurposed antibodies or their target-binding fragments (if desired, together with other ingredients) can also be encapsulated in hard or soft shell gelatin capsules, compressed into tablets or directly incorporated into the diet of the subject. For oral therapeutic administration, the original antibodies and / or synthetic molecules (or portions thereof) of the present disclosure, the repurposed antibodies or their target-binding fragments can be incorporated with excipients and used in the form of ingestible tablets, buccal tablets, lozenges, capsules, elixirs, suspensions, syrups, wafers, etc. In order to administer the original antibodies and / or synthetic molecules (or portions thereof) of the present disclosure, the repurposed antibodies of the present disclosure or their target-binding fragments by a non-parenteral route, it may be necessary to coat the antibody or its target-binding fragment with a material or co-administer the antibody or its target-binding fragment with a material to prevent its inactivation.

[0211] In certain embodiments, the therapeutically effective amount of the repurposed antibody is determined to be the amount provided in the package insert provided with the original antibody. The term package insert refers to the instructions typically included in the commercial package of a drug approved by the FDA or a similar regulatory agency outside the United States, which contains information about the use, dosage, administration, contraindications and / or warnings of such drug.

[0212] A repurposed antibody or a target-binding fragment thereof can be administered in an amount related to the weight of a patient in need of such treatment. In one embodiment, a repurposed antibody or a target-binding fragment thereof can be administered in the following amounts: 0.1 mg / kg to about 50 mg / kg, 0.1 mg / kg to about 40 mg / kg, 0.1 mg / kg to about 30 mg / kg, 0.1 mg / kg to about 25 mg / kg, 0.1 mg / kg to about 20 mg / kg, 0.1 mg / kg to about 15 mg / kg, 0.1 mg / kg to about 10 mg / kg, 0.1 mg / kg to about 7.5 mg / kg, 0.1 mg / kg to about 5 mg / kg, 0.1 mg / kg to about 2.5 mg / kg, or about 0.1 mg / kg to about 1 mg / kg. A repurposed antibody or a target-binding fragment thereof can be administered in the following amounts: 0.5 mg / kg to about 50 mg / kg, 0.5 mg / kg to about 40 mg / kg, 0.5 mg / kg to about 30 mg / kg, 0.5 mg / kg to about 25 mg / kg, 0.5 mg / kg to about 20 mg / kg, 0.5 mg / kg to about 15 mg / kg, 0.5 mg / kg to about 10 mg / kg, 0.5 mg / kg to about 7.5 mg / kg, 0.5 mg / kg to about 5 mg / kg, 0.5 mg / kg to about 2.5 mg / kg, or about 0.5 mg / kg to about 1 mg / kg. A repurposed antibody or a target-binding fragment thereof can be administered in an amount of about 0.5 mg / kg to about 5 mg / kg or about 0.1 mg / kg to about 10 mg / kg. A repurposed antibody or a target-binding fragment thereof can be administered in an amount of about 0.1 mg / kg to about 20 mg / kg or about 0.1 mg / kg to about 30 mg / kg.

[0213] The reutilized antibody or its target-binding fragment can be administered in an amount of approximately: 1 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 75 mg, 80 mg, 90 mg, 100 mg, 150 mg, or 200 mg. The reutilized antibody or its target-binding fragment can be administered in an amount of approximately: 250 mg, 300 mg, 400 mg, 500 mg, 600 mg, 700 mg, 800 mg, 900 mg, 1000 mg, 1100 mg, 1200 mg, 1300 mg, 1400 mg, 1500 mg, 1600 mg, 1700 mg, 1800 mg, 1900 mg, or 2000 mg. The reutilized antibody or its target-binding fragment can be administered in an amount of approximately 1000 mg to approximately 2000 mg. The reutilized antibody or its target-binding fragment can be administered in an amount of approximately: 1 mg to approximately 10 mg, 10 mg to approximately 20 mg, 25 mg to approximately 50 mg, 30 mg to approximately 60 mg, 40 mg to approximately 50 mg, 50 mg to approximately 100 mg, 75 mg to approximately 150 mg, 100 mg to approximately 200 mg, 200 mg to approximately 500 mg, 500 mg to approximately 1000 mg, 1000 mg to approximately 1200 mg, 1000 mg to approximately 1500 mg, 1200 mg to approximately 1500 mg, or 1500 to approximately 2000 mg.

[0214] The reutilized antibody or its target-binding fragment can be administered in an amount of approximately 0.1 mg / mL, 0.5 mg / mL, 1 mg / mL, 2 mg / mL, 3 mg / mL, 4 mg / mL, 5 mg / mL, 6 mg / mL, 7 mg / mL, 8 mg / mL, 9 mg / mL, 10 mg / mL, 15 mg / mL, 20 mg / mL, 25 mg / mL, 30 mg / mL, 40 mg / mL, 50 mg / mL, 60 mg / mL, 70 mg / mL, 80 mg / mL, 90 mg / mL, 100 mg / mL, 150 mg / mL, 200 mg / mL, 250 mg / mL, 300 mg / mL, 400 mg / mL, or 500 mg / mL. In one embodiment, the reutilized antibody or its target-binding fragment is present in the combination in an amount of approximately: 1 mg / mL to approximately 10 mg / mL, 5 mg / mL to approximately 10 mg / mL, 5 mg / mL to approximately 15 mg / mL, 10 mg / mL to approximately 25 mg / mL; 20 mg / mL to approximately 30 mg / mL; 25 mg / mL to approximately 50 mg / mL, or 50 mg / mL to approximately 100 mg / mL.

[0215] For example, the first molecule and / or the second molecule can be administered continuously or intermittently (such as, BIW), for example, once a day (QD), twice a day (BID), once a week (QW), twice a week (BIW), three times a week (TIW), or once a month (QM), for 3 months, and then resumed after one month. In one embodiment, the first molecule and / or the second molecule can be administered BID. The first molecule and / or the second molecule can be administered TIW. In some cases, the first molecule and / or the second molecule are administered 2 to 3 times per week. In another embodiment, the first molecule and / or the second molecule are administered QD. The compound can be administered QD for about: 1 day to about 7 days, 1 day to about 14 days, 1 day to about 21 days, 1 day to about 28 days, or administered daily until disease progression or unacceptable toxicity occurs. The administration of the first molecule and / or the second molecule and / or the synthetic compound may depend in part on the patient's tolerance, and the higher the tolerance, the more times or more frequently the administration is. Alternatively, when the patient has poor tolerance to the first molecule and / or the second molecule and / or the synthetic compound, the dosage of the compound can be reduced or the administration frequency can be decreased. The compound of formula I can be administered according to any regimen described herein.

[0216] In one embodiment, the first molecule and / or the second molecule and / or the synthetic compound may be administered in an amount of approximately: 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 60 mg, 70 mg, 80 mg, 85 mg, 90 mg, 100 mg, 125 mg, 150 mg, 175 mg, or 200 mg, QD. In one embodiment, the first molecule and / or the second molecule and / or the synthetic compound may be administered in an amount of approximately: 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 60 mg, 70 mg, 80 mg, 85 mg, 90 mg, 100 mg, 125 mg, 150 mg, 175 mg, or 200 mg, BIW. In one embodiment, the first molecule and / or the second molecule and / or the synthetic compound may be administered in an amount of approximately: 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 60 mg, 70 mg, 80 mg, 85 mg, 90 mg, 100 mg, 125 mg, 150 mg, 175 mg, or 200 mg, TIW. In one embodiment, the first molecule and / or the second molecule and / or the synthetic compound may be administered in an amount of approximately: 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 60 mg, 70 mg, 80 mg, 85 mg, 90 mg, 100 mg, 125 mg, 150 mg, 175 mg, or 200 mg, QW. In one embodiment, the first molecule and / or the second molecule and / or the synthetic compound may be administered in an amount of approximately: 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 60 mg, 70 mg, 80 mg, 85 mg, 90 mg, 100 mg, 125 mg, 150 mg, 175 mg, or 200 mg, Q2W. In one embodiment, the first molecule and / or the second molecule and / or the synthetic compound may be administered in an amount of approximately 5 mg or approximately 10 mg, QD. In one embodiment, the first molecule and / or the second molecule and / or the synthetic compound may be administered in an amount of approximately 5 mg or approximately 10 mg, BIW. In one embodiment, the first molecule and / or the second molecule and / or the synthetic compound may be administered in an amount of approximately 5 mg or approximately 10 mg, TIW.In one embodiment, the first molecule and / or the second molecule and / or the synthetic compound, QW, may be administered in an amount of about 5 mg or about 10 mg. In one embodiment, the first molecule and / or the second molecule and / or the synthetic compound, Q2W, may be administered in an amount of about 5 mg or about 10 mg. The administration of the first molecule and / or the second molecule and / or the synthetic compound may be continuous. The administration of the first molecule and / or the second molecule and / or the synthetic compound may be intermittent.

[0217] In one embodiment, the first molecule and / or the second molecule and / or the synthetic compound, QD, may be administered in an amount of about: 1 mg to about 10 mg, 1 mg to about 25 mg, 1 mg to about 50 mg, 5 mg to about 10 mg, 5 mg to about 25 mg, 5 mg to about 50 mg, 10 mg to about 25 mg, 10 mg to about 50 mg, 50 mg to about 100 mg, or 100 mg to about 200 mg. In one embodiment, the first molecule and / or the second molecule and / or the synthetic compound, BIW, may be administered in an amount of about: 1 mg to about 10 mg, 1 mg to about 25 mg, 1 mg to about 50 mg, 5 mg to about 10 mg, 5 mg to about 25 mg, 5 mg to about 50 mg, 10 mg to about 25 mg, 10 mg to about 50 mg, 50 mg to about 100 mg, or 100 mg to about 200 mg. In one embodiment, the first molecule and / or the second molecule and / or the synthetic compound, TIW, may be administered in an amount of about: 1 mg to about 10 mg, 1 mg to about 25 mg, 1 mg to about 50 mg, 5 mg to about 10 mg, 5 mg to about 25 mg, 5 mg to about 50 mg, 10 mg to about 25 mg, 10 mg to about 50 mg, 50 mg to about 100 mg, or 100 mg to about 200 mg. In one embodiment, the first molecule and / or the second molecule and / or the synthetic compound, QW, may be administered in an amount of about: 1 mg to about 10 mg, 1 mg to about 25 mg, 1 mg to about 50 mg, 5 mg to about 10 mg, 5 mg to about 25 mg, 5 mg to about 50 mg, 10 mg to about 25 mg, 10 mg to about 50 mg, 50 mg to about 100 mg, or 100 mg to about 200 mg. In one embodiment, the first molecule and / or the second molecule and / or the synthetic compound, Q2W, may be administered in an amount of about: 1 mg to about 10 mg, 1 mg to about 25 mg, 1 mg to about 50 mg, 5 mg to about 10 mg, 5 mg to about 25 mg, 5 mg to about 50 mg, 10 mg to about 25 mg, 10 mg to about 50 mg, 50 mg to about 100 mg, or 100 mg to about 200 mg. The administration of the first molecule and / or the second molecule and / or the synthetic compound may be continuous. The administration of the first molecule and / or the second molecule and / or the synthetic compound may be intermittent.

[0218] In one embodiment, the first molecule and / or the second molecule and / or the synthetic compound can be administered in an amount of approximately: 0.0001 mg / kg to about 200 mg / kg, 0.001 mg / kg to about 200 mg / kg, 0.01 mg / kg to about 200 mg / kg, 0.01 mg / kg to about 150 mg / kg, 0.01 mg / kg to about 100 mg / kg, 0.01 mg / kg to about 50 mg / kg, 0.01 mg / kg to about 25 mg / kg, 0.01 mg / kg to about 10 mg / kg, or 0.01 mg / kg to about 5 mg / kg, QD. In one embodiment, the first molecule and / or the second molecule and / or the synthetic compound can be administered in an amount of approximately: 0.0001 mg / kg to about 200 mg / kg, 0.001 mg / kg to about 200 mg / kg, 0.5 mg / kg to about 200 mg / kg, 0.5 mg / kg to about 150 mg / kg, 0.5 mg / kg to about 100 mg / kg, 0.5 mg / kg to about 50 mg / kg, 0.5 mg / kg to about 25 mg / kg, 0.5 mg / kg to about 10 mg / kg or 0.5 mg / kg to about 5 mg / kg, BIW. In one embodiment, the first molecule and / or the second molecule and / or the synthetic compound can be administered in an amount of approximately: 0.0001 mg / kg to about 200 mg / kg, 0.001 mg / kg to about 200 mg / kg, 0.5 mg / kg to about 200 mg / kg, 0.5 mg / kg to about 150 mg / kg, 0.5 mg / kg to about 100 mg / kg, 0.5 mg / kg to about 50 mg / kg, 0.5 mg / kg to about 25 mg / kg, 0.5 mg / kg to about 10 mg / kg or 0.5 mg / kg to about 5 mg / kg, TIW.In one embodiment, the first molecule and / or the second molecule and / or the synthetic compound may be administered in an amount of about: 0.0001 mg / kg to about 200 mg / kg, 0.001 mg / kg to about 200 mg / kg, 0.5 mg / kg to about 200 mg / kg, 0.5 mg / kg to about 150 mg / kg, 0.5 mg / kg to about 100 mg / kg, 0.5 mg / kg to about 50 mg / kg, 0.5 mg / kg to about 25 mg / kg, 0.5 mg / kg to about 10 mg / kg or 0.5 mg / kg to about 5 mg / kg, QW. In one embodiment, the first molecule and / or the second molecule and / or the synthetic compound may be administered in an amount of about: 0.0001 mg / kg to about 200 mg / kg, 0.001 mg / kg to about 200 mg / kg, 0.5 mg / kg to about 200 mg / kg, 0.5 mg / kg to about 150 mg / kg, 0.5 mg / kg to about 100 mg / kg, 0.5 mg / kg to about 50 mg / kg, 0.5 mg / kg to about 25 mg / kg, 0.5 mg / kg to about 10 mg / kg or 0.5 mg / kg to about 5 mg / kg, Q2W. In one example, the first molecule and / or the second molecule and / or the synthetic compound may be administered in an amount of about 15 mg / kg to about 75 mg / kg, QD. In another example, the first molecule and / or the second molecule and / or the synthetic compound may be administered in an amount of about 20 mg / kg to about 50 mg / kg. In yet another example, the first molecule and / or the second molecule and / or the synthetic compound may be administered in an amount of about 0.001 mg / kg, 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, 1 mg / kg, 2 mg / kg, 3 mg / kg, 4 mg / kg, 5 mg / kg, 10 mg / kg, 15 mg / kg, 20 mg / kg, 25 mg / kg, 30 mg / kg, 40 mg / kg, 50 mg / kg, 60 mg / kg, 70 mg / kg, 80 mg / kg, 90 mg / kg, 100 mg / kg, 125 mg / kg, 150 mg / kg, 175 mg / kg or 200 mg / kg. The administration of the first molecule and / or the second molecule and / or the synthetic compound may be continuous. The administration of the first molecule and / or the second molecule and / or the synthetic compound may be intermittent.

[0219] In one embodiment, the first molecule and / or the second molecule and / or the synthetic compound may be administered in an amount of about: 1 mg / kg to about 200 mg / kg, 1 mg / kg to about 150 mg / kg, 1 mg / kg to about 100 mg / kg, 1 mg / kg to about 50 mg / kg, 1 mg / kg to about 25 mg / kg, 1 mg / kg to about 10 mg / kg, or 1 mg / kg to about 5 mg / kg, QD. In one embodiment, the first molecule and / or the second molecule and / or the synthetic compound may be administered in an amount of about: 1 mg / kg to about 200 mg / kg, 1 mg / kg to about 150 mg / kg, 1 mg / kg to about 100 mg / kg, 1 mg / kg to about 50 mg / kg, 1 mg / kg to about 25 mg / kg, 1 mg / kg to about 10 mg / kg, or 1 mg / kg to about 5 mg / kg, BIW. In one embodiment, the first molecule and / or the second molecule and / or the synthetic compound may be administered in an amount of about: 1 mg / kg to about 200 mg / kg, 1 mg / kg to about 150 mg / kg, 1 mg / kg to about 100 mg / kg, 1 mg / kg to about 50 mg / kg, 1 mg / kg to about 25 mg / kg, 1 mg / kg to about 10 mg / kg, or 1 mg / kg to about 5 mg / kg, TIW. In one embodiment, the first molecule and / or the second molecule and / or the synthetic compound may be administered in an amount of about: 1 mg / kg to about 200 mg / kg, 1 mg / kg to about 150 mg / kg, 1 mg / kg to about 100 mg / kg, 1 mg / kg to about 50 mg / kg, 1 mg / kg to about 25 mg / kg, 1 mg / kg to about 10 mg / kg, or 1 mg / kg to about 5 mg / kg, QW. In one embodiment, the first molecule and / or the second molecule and / or the synthetic compound may be administered in an amount of about: 1 mg / kg to about 200 mg / kg, 1 mg / kg to about 150 mg / kg, 1 mg / kg to about 100 mg / kg, 1 mg / kg to about 50 mg / kg, 1 mg / kg to about 25 mg / kg, 1 mg / kg to about 10 mg / kg, or 1 mg / kg to about 5 mg / kg, Q2W. In one example, the first molecule and / or the second molecule and / or the synthetic compound may be administered in an amount of about 15 mg / kg to about 75 mg / kg, QD. In another example, the first molecule and / or the second molecule and / or the synthetic compound may be administered in an amount of about 20 mg / kg to about 50 mg / kg.In yet another example, the first molecule and / or the second molecule and / or the synthetic compound can be administered in an amount of about 0.001 mg / kg, 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.5 mg / kg, 1 mg / kg, 2 mg / kg, 3 mg / kg, 4 mg / kg, 5 mg / kg, 10 mg / kg, 15 mg / kg, 20 mg / kg, 25 mg / kg, 30 mg / kg, 40 mg / kg, 50 mg / kg, 60 mg / kg, 70 mg / kg, 80 mg / kg, 90 mg / kg, 100 mg / kg, 125 mg / kg, 150 mg / kg, 175 mg / kg, or 200 mg / kg. The administration of the first molecule and / or the second molecule and / or the synthetic compound can be continuous. The administration of the first molecule and / or the second molecule and / or the synthetic compound can be intermittent.

[0220] The original antibody and / or the synthetic molecule (or a portion thereof) of the present disclosure, a repurposed antibody or a target-binding fragment thereof, or any combination described herein can be administered in a regimen. The regimen can be configured to provide a therapeutically effective amount of the original antibody and / or the synthetic molecule (or a portion thereof) of the present disclosure, a repurposed antibody or a target-binding fragment thereof, or any combination described herein over a predetermined period of time (e.g., the administration time). The regimen can be configured to limit or prevent side effects or adverse complications of the components of the original antibody and / or the synthetic molecule (or a portion thereof) of the present disclosure, a repurposed antibody or a target-binding fragment thereof, or any combination described herein. The regimen can be configured to result in an increased effect (e.g., synergy) of the two therapies in combination. A regimen for treating cancer can include administration for any number of days and can be repeated as needed. The administration period can be interrupted by a rest period, where the rest period includes not administering at least one therapy. For example, the regimen can include an administration period that includes 2, 3, 5, 7, 10, 15, 21, 28, or more days. These cycles can be repeated. For example, the regimen can include a set number of days as described above, where the regimen is repeated 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or more times.

[0221] In other embodiments, the regimen can include a rest period of at least 1, 2, 3, 5, 7, 10, or more days, where at least one therapy is no longer administered to the patient. For example, the rest period can be determined by monitoring the patient's response to the original antibody and / or the synthetic molecule (or a portion thereof) of the present disclosure, a repurposed antibody or a target-binding fragment thereof, or any combination described herein or by measuring the efficacy of the treatment. The rest period can apply to a single therapy such that only one therapy in the combination described herein is stopped during the rest period, but the other therapies are still administered. The rest period can apply to all therapies administered to the subject such that the subject does not receive any therapy for a set period of time during the rest period.

[0222] Combined treatment

[0223] In some embodiments, the original antibody and / or a synthetic molecule (or portion thereof) of the present disclosure, a repurposed antibody, or a target-binding fragment thereof may be formulated with another therapeutic agent. The agent can be any agent so long as the agent is also useful for the indication for which the repurposed antibody is directed.

[0224] In some embodiments, the original antibody and / or a synthetic molecule (or portion thereof) of the present disclosure, a repurposed antibody, or a target-binding fragment thereof may be co-administered with another therapeutic agent or treatment (such as radiation). In some embodiments, the therapeutic agent or treatment is administered before, simultaneously with, and / or after the original antibody and / or the synthetic molecule (or portion thereof) of the present disclosure, the repurposed antibody, or a target-binding fragment thereof. As used herein, "co-administered" means delivering two (or more) different treatments (e.g., a repurposed antibody and an antibiotic) to a subject during the course of the subject's illness, e.g., after the subject has been diagnosed with a disease and before the disease has been cured or eliminated or treatment has been discontinued for other reasons. In some embodiments, at the start of delivery of the second treatment, delivery of the first treatment is still ongoing, such that there is overlap in administration. This is sometimes referred to herein as "simultaneous" or "concurrent" delivery. In other embodiments, delivery of the other treatment is initiated after delivery of the first treatment has terminated. In embodiments of either of these two cases, the treatments are more effective due to the combined administration. For example, the second treatment is more effective (e.g., the same effect is observed with less of the second treatment, or the second treatment alleviates or reduces symptoms to a greater extent) than would be observed if only the second treatment were administered without the first treatment, or vice versa for the first treatment. In some embodiments, symptoms (e.g., toxicity caused by administration of a repurposed antibody) or other parameters associated with the disorder are alleviated or reduced or decreased to a greater extent than would be observed with the delivery of one treatment without the other. The effects of the two treatments may be partially additive, fully additive, or more than additive. Delivery may allow the effect of delivering the first treatment to still be detected during delivery of the second treatment. The molecules of the present disclosure and the additional therapeutic agent may be administered simultaneously, in the same or separate compositions, or sequentially. In one embodiment, for sequential administration, the repurposed antibody or other molecule described herein may be administered first, followed by the therapeutic agent, or the order of administration may be reversed. The repurposed antibody or any other molecule described herein may be administered during active disease or during a remission or less active disease period. The repurposed antibody or any other molecule described herein may be administered before, concurrently with, after a treatment, or during a remission of a disease (e.g., cancer).

[0225] In one embodiment, the other therapeutic agents include agents that have the following effects: antibiotics, antiviral agents, anti-inflammatory agents, cytokines, hematopoietic growth factors, anti-cancer agents (including chemotherapeutic agents), immunomodulators, immunosuppressants, steroids (e.g., corticosteroids) or pharmacologically active derivatives thereof, therapeutic antibodies, vitamins, calcium or calcium supplements.

[0226] In some embodiments, the therapeutic agent is another antibody. In some embodiments, the other antibody is selected from those listed in Table 1.

[0227] [In one embodiment, the therapeutic agent is selected from amikacin, amoxicillin, amoxicillin-clavulanic acid, amphotericin-B, ampicillin, ampicillin-sulbactam, apramycin, azithromycin, aztreonam, bacitracin, benzylpenicillin, caspofungin, cefaclor, cefadroxil, cephalexin, cephalosporin Thiophene, cefazolin, cefdinir, cefepime, cefixime, cefmenoxime, cefoperazone, cefoperazone-sulbactam, ceftriaxone, cefotaxime, cefoxitin, cefepime, cefpodoxime, cefpodoxime-clavulanic acid, cefpodoxime-sulbactam, cefbroxil, cefquinol, ceftazidime, cefbutin, ceftiofur, ceftobiprole, ceftriaxone, cefuroxime, chloramphenicol, florfenicol, ciprofloxacin, clarithromycin , clinafloxacin, clindamycin, cloxacillin, colistin, trimethoprim / sulfamethoxazole, dalbavancin, dalfopristin / quinopristin, daptomycin, dibekacin, dicloxacillin, doripenem, doxycycline, enrofloxacin, ertapenem, erythromycin, flucloxacillin, fluconazole, flucytosine, fosfomycin, fusidic acid, gareoxacin, gatifloxacin, gemifloxacin, gentamicin, imipenem Nan, itraconazole, kanamycin, ketoconazole, levofloxacin, lincomycin, linezolid, clocarbazine, mecillin (aminocillin), meropenem, metronidazole, mezlocillin, mezlocillin sulbactam, minocycline, moxifloxacin, mupirocin, nalidixic acid, neomycin, netilmicin, nitrofurantoin, norfloxacin, ofloxacin, oxacillin, pefloxacin, penicillin V (Penicillin V), piperacillin, piperacillin-sulbactam, piperacillin-tazobactam, rifampicin, roxithromycin, sparfloxacin, spectinomycin, spiramycin, streptomycin, sulbactam, sulfamethoxazole, teicoplanin, telavancin, telithromycin, temocillin, tetracycline, ticarcillin, ticarcillin-clavulanic acid, tigecycline, tobramycin, trimethoprim, trovafloxacin, tylosin, vancomycin, virginiamycin, voriconazole and combinations thereof.

[0228] In one embodiment, the antiviral agent is selected from remdesivir, oseltamivir phosphate, zanamivir, peramivir, baloxavir marboxil, darunavir, atazanavir, ritonavir, acyclovir, valacyclovir, valganciclovir, tenofovir, raltegravir, virus attachment inhibitors, virus entry inhibitors, uncoating inhibitors, protease inhibitors, polymerase inhibitors, nucleoside and nucleotide reverse transcriptase inhibitors, non-nucleoside reverse transcriptase inhibitors, integrase inhibitors; nucleoside analogs (such as zidovudine, acyclovir, ganciclovir, vidarabine, idoxuridine, trifluridine and ribavirin), foscarnet, amantadine, peramivir, rimantadine, saquinavir, indinavir, ritonavir, α-interferon and other interferons, AZT, t-705, zanamivir and oseltamivir Other antiviral agents include influenza virus vaccines, such as (Glaxo SmithKline), (MedlmmuneVaccines), (Chiron Corporation), (Glaxo SmithKline), (CSL Biotherapies Inc.), (Novartis), (Aventis Pasteur), remdesivir, lopinavir, favipiravir, darunavir, oseltamivir, umifenovir, novaferon; immunomodulatory drugs selected from immunoglobulins, monoclonal antibodies; glucocorticoids; anti-inflammatory drugs selected from leflunomide, colchicine, naproxen, piclidenoson; cardiovascular drugs selected from ACE-2, ACE inhibitors, ARB, angiotensin; chloroquine; hydroxychloroquine; avitamin, azithromycin; itraconazole; zidovudine, acyclovir, ganciclovir, vidarabine, idoxuridine, trifluridine, ribavirin, foscarnet, amantadine, peramivir, rimantadine, saquinavir, indinavir, ritonavir, α-interferon, AZT, t-705, zanamivir, oseltamivir; vitamin D; zinc; and combinations thereof.

[0229] In one embodiment, the therapeutic agent is an anti-SARS-CoV-2 vaccine selected from the group consisting of: intranasal SARS-CoV-2 vaccine (Altimmune), INO-4800 (Inovio Pharma) and Beijing Advaccine Biotechnology Company, APN01 (APEIRON Biologics), mRNA-1273 vaccine (Moderna and Vaccine Research Center), nucleoside-modified mRNA BNT162b2 Tozinameran (INN) (Pfizer-BioNTech), adenovirus-based vaccine AZD1222 (recombinant ChAdOx1 adenovirus vector encoding the SARS-CoV-2 spike protein antigen; Oxford-AstraZeneca), Covishield (ChAdOx1_nCoV19) recombinant ChAdOx1 adenovirus vector encoding the SARS-CoV-2 spike protein antigen (Serum Institute of India), SARS-CoV-2 vaccine (Vero cells), inactivated vaccine (lnCoV) (Sinopharm / BIBP), SARS-CoV-2 vaccine (Vero cells), inactivated vaccine (Sinovac), Ad26.COV2.S-recombinant, replication-incompetent adenovirus type 26 (Ad26) vector vaccine (encoding the spike (S) protein of SARS-CoV-2) (Janssen Pharmaceuticals Companies of Johnson & Johnson), Covid-19 vaccine based on Sputnik V human adenovirus vector (The Gamaleya National Center), Ad5-nCoV recombinant novel coronavirus vaccine (adenovirus type 5 vector) (CanSinoBIO), EpiVacCorona peptide antigen vaccine (Vector State Research Centre of Virology and Biotechnology, Russia), recombinant novel coronavirus vaccine (CHO) (ZhifeiLongcom, China), SARS-CoV-2 vaccine, inactivated vaccine (Vero cells) (IMBCAMS, China), inactivated SARS-CoV-2 vaccine (Vero cells) (Sinopharm / WIBP), avian coronavirus infectious bronchitis virus (IBV) vaccine (MIGDAL Research Institute), modified vaccinia virus vaccine TNX-1800 (Tonix Pharmaceuticals), recombinant subunit vaccine based on the SARS-CoV-2 coronavirus trimeric S protein (S-Trimer) (Clover Pharmaceuticals), oral recombinant coronavirus vaccine (Vaxart), linear DNA vaccine based on (i) the entire spike gene of the coronavirus or (ii) the protein antigenic part of the coronavirus (Applied DNA Sciences and Takis Biotech), SARS-Cov-2 coronavirus vaccine NVX-CoV2373 (Novavax), SARS-Cov-2 coronavirus vaccine NVX-CoV2373 (Novavax), intramuscular vaccine INO-4700 (GLS-5300) (Inovio Pharma and GeneOne Life Science) and their combinations.

[0230] In one embodiment, the therapeutic agent is a SARS-CoV-2 RNA polymerase inhibitor, serine protease inhibitor, cysteine protease inhibitor, galidesivir, remdesivir, hydroxychloroquine, chloroquine, irbesartan, toremifene, camphor, maprotiline, mesalazine, mercaptopurine, nafamostat, parabens, sirolimus, carvedilol, actinomycin, melatonin, quinacrine, eplerenone, enolin, hydroxymetholone, ENU2000, azithromycin, lopinavir / ritonavir, umifenovir, cytovene, ganciclovir, trisodium phosphate, ribavirin, interferon, d4T, ddl, AZT, amantadine, rimantadine, acyclovir, foscarnet, laninamivir, oseltamivir, zanamivir, favipiravir, baloxavir marboxil, and peramivir, or a combination thereof.

[0231] In one embodiment, the therapeutic agent is a "checkpoint inhibitor". The term "checkpoint inhibitor" refers to a molecule that fully or partially reduces, inhibits, interferes with, or modulates one or more checkpoint proteins. Without being limited to a particular theory, checkpoint proteins regulate T cell activation or function. A variety of checkpoint proteins are known, such as CTLA-4 and its ligands CD80 and CD86; and PD-1 and its ligands PD-L1 and PD-L2. These proteins appear to be responsible for the co-stimulatory or inhibitory interactions of T cell responses. Immune checkpoint proteins appear to regulate and maintain self-tolerance as well as the duration and magnitude of physiological immune responses.

[0232] In one embodiment, the therapeutic agent is one or more immune cells (e.g., modified immune cells) that express one or more chimeric antigen receptors (CARs) on their surface. Generally, a CAR comprises an extracellular domain (e.g., an antigen-binding protein) from a first protein, a transmembrane domain, and an intracellular signaling domain. In certain embodiments, once the extracellular domain binds to a target protein (such as a tumor-associated antigen (TAA) or a tumor-specific antigen (TSA)), a signal that activates the immune cell is generated through the intracellular signaling domain, e.g., to target and kill cells that express the target protein. In some embodiments, the therapeutic agent can be together with one or more immune cells (e.g., modified immune cells) that express one or more chimeric antigen receptors (CARs) on their surface. Generally, a CAR comprises an extracellular domain (e.g., an antigen-binding protein) from a first protein, a transmembrane domain, and an intracellular signaling domain. In certain embodiments, once the extracellular domain binds to a target protein (such as a tumor-associated antigen (TAA) or a tumor-specific antigen (TSA)), a signal that activates the immune cell is generated through the intracellular signaling domain, e.g., to target and kill cells that express the target protein. The modified immune cells expressing CAR can be, for example, T lymphocytes (T cells, e.g., CD4+ T cells or CD8+ T cells), cytotoxic lymphocytes (CTLs), or natural killer (NK) cells. The T lymphocytes used in the compositions and methods provided herein can be naive T lymphocytes or MHC-restricted T lymphocytes. In certain embodiments, the T lymphocytes are tumor-infiltrating lymphocytes (TILs). In certain embodiments, T lymphocytes have been isolated from a tumor biopsy or T lymphocytes isolated from a tumor biopsy have been expanded. In certain other embodiments, T cells have been isolated from peripheral blood, cord blood, or lymph or T lymphocytes isolated from peripheral blood, cord blood, or lymph have been expanded. Conventional methods well known in the art can be used to isolate the immune cells to be used for generating the modified immune cells expressing CAR, such as blood collection, followed by apheresis and optionally antibody-mediated cell separation or sorting. The modified immune cells are preferably autologous to the individual to whom the modified immune cells are to be administered. In certain other embodiments, the modified immune cells are allogeneic to the individual to whom the modified immune cells are to be administered. When allogeneic T lymphocytes or NK cells are used to prepare modified T lymphocytes, T lymphocytes or NK cells that can reduce the likelihood of an individual developing graft-versus-host disease (GVHD) are preferably selected. For example, in certain embodiments, virus-specific T lymphocytes are selected to prepare modified T lymphocytes; it is expected that the original binding ability of such lymphocytes to any recipient antigen is significantly reduced, thereby reducing the likelihood of their activation.In some embodiments, receptor-mediated allogeneic T lymphocyte rejection can be reduced by co-administering to the host one or more immunosuppressive agents (such as cyclosporine, tacrolimus, sirolimus, cyclophosphamide, etc.).

[0233] In one embodiment, the chemotherapeutic agent is selected from acevine; aclarubicin; acridazone hydrochloride; actinonin; adozenastine; aldesleukin; altretamine; ambomycin; ametantrone acetate; amsacrine; anastrozole; anthramycin; asparaginase; aspirin; asitopride; azomycin; batimastat; benzpiperidine; bicalutamide; bisantrene hydrochloride; bisnafide dimethanesulfonate; bizelesin; bleomycin sulfate; brequinar sodium; bromopirimine; busulfan; actinomycin; caroxazone; casimamide; carbetimer; carboplatin; carmustine; carubicin hydrochloride; cazelesin; cedefadol; celecoxib (a COX-2 inhibitor); chlorambucil; cirolemycin; cisplatin; cladribine; clofarabine; creanoline mesylate; cyclophosphamide; cytarabine; dacarbazine; actinomycin; daunorubicin hydrochloride; decitabine; dicarboplatin; dideoxyguanosine; dideoxyguanosine mesylate; diquone; docetaxel; doxorubicin; doxorubicin hydrochloride; droloxifene; droloxifene citrate; droloxifene propionate; duazomycin; edatrexate; eflornithine hydrochloride; elsamitrucin; enloplatin; enprostil; epipropidine; epirubicin hydrochloride; erbucin; esorubicin hydrochloride; estramustine; estramustine phosphate sodium; etanidazole; etoposide; etoposide phosphate; etoprine; fazarabine; fenretinide; floxuridine; fludarabine phosphate; fluorouracil; flucytosine; fosquidone; fostriecin sodium; gemcitabine; gemcitabine hydrochloride; hydroxyurea; idarubicin hydrochloride; ifosfamide; imofosine; iproplatin; irinotecan; irinotecan hydrochloride; lanreotide acetate; letrozole; leuprorelin acetate; riluzole hydrochloride; lometrexol sodium; lomustine; losoxantrone hydrochloride; masoprocol; maytansine; mechlorethamine hydrochloride; megestrol acetate; melengestrol acetate; melphalan; menogaril; mercaptopurine; methotrexate; methotrexate sodium; metoprine; meturedepa; mitodiamide; mitomycin; mitoromine; mitogillin; mitomycin; mitotane; mitoxantrone hydrochloride; mycophenolic acid; nocodazole; nogalamycin; omacetaxine; omarplatin; oxilan; paclitaxel; pegaspargase; pellimycin; pentamustine; peplomycin sulfate; peplosamide; pipobroman; piposulfan; pirarubicin hydrochloride; plicamycin; promestriene; porfimer sodium; porfiromycin; prednimustine; procarbazine hydrochloride; puromycin; puromycin hydrochloride; pyrazofurin; riboprine; safingol; safingol hydrochloride; semustine; simtrazine; sorafenib; strontium pamidronate; sparmomycin; spirogermanium hydrochloride; spiro mustard; spiroplatin; streptozocin; streptozocin; sulfofenox; talisomycin; ticagalan sodium; taxotere; tegafur; teloxantrone hydrochloride; temoporfin; teniposide; troxipide; testolactone; tioguanine; thiotepa; thiazofurin; tirapazamine; toremifene citrate; tritium acetate; triciribine phosphate; trimetrexate; trimetrexate glucuronate; triptorelin; tubulozole hydrochloride; uracil mustard; uracil pipobroman; vapreotide; verteporfin;Vinblastine Sulfate; Vincristine Sulfate; Vindesine; Vindesine Sulfate; Vinfluprine Sulfate; Vinblastine Sulfate; Vinblastine Sulfate; Vinblastine Sulfate; Vinorelbine Tartrate; Vindolidine Sulfate; Vinleurosine Sulfate; Fosrozole; Zeniplatin; Zinostatin; and Zorubicin Hydrochloride.;

[0234] Other anticancer drugs included in the method of the present invention include, but are not limited to: 20-epi-1,25-dihydroxyvitamin D3; 5-ethynyluracil; abiraterone; aclarubicin; acylfulvene; nandrolone decanoate; adozenecin; aldesleukin; ALL-TK antagonist; altretamine; ametycin; amidox; amifostine; aminolevulinic acid; amrubicin; ametantrone; anagrelide; anastrozole; andrographolide; angiogenesis inhibitor; antagonist D; antagonist G; antarelix; anti-dorsalizing morphogenetic protein-1; antiandrogen, prostate cancer; antiestrogen; antiangiogenin; antisense oligonucleotide; aphidicolinglycinate; apoptosis gene regulator; apoptosis regulator; apurinic acid; ara-CDP-DL-PTBA; arginine deaminase; asulacrine; atamestane; atrimustine; aspirin 1; aspirin 2; aspirin 3; azasetron; azatoxin; azatyrosine; baccatin III derivative; balanol; batimastat; BCR / ABL antagonist; benzochlorin; benzoylstaricin; β-lactam derivative; β-allopurinol; betaclarithromycin B; betulinic acid; bFGF inhibitor; bicalutamide; bisantrene; bisaziridinylspermine; binaphthyl; bitriptolide A; bizelesin; bufuralol; bromopirimine; budotitane; buthionine sulfoximine; calcipotriol; carboprost C; camptothecin derivative; capecitabine; carboxyamidotriazole; carboxyamide triazole; CaRest M3; CARN 700; cartilage-derived inhibitor; cazelesin; casein kinase inhibitor (ICOS); castanospermine; cecropin B; cetrorelix; chloramphenicol; chloroquinoline sulfonamide; cicaprost; cis-porphyrin; cladribine; clomifene analogue; clotrimazole; collimycin A; collimycin B; combretastatin A4; combretastatin analogue; conagenin; crambescidin 816; crisnatol; cryptophycin 8; cryptophycin A derivative; curacin A; cyclopentanthraquinone; carboplatin; cypemycin; Ara-C octadecanoate; cytolytic factor; cytostatin; daclizumab; decitabine; dehydroepiandrosterone B; deslorelin; dexifosfamide; dexrazoxane; diltiazem; diquat; didemnin B; dioxide; diethylnorspermine; dihydro-5-azacytidine; 9-dihydrotaxol; doxorubicin; droloxifene; dronabinol; docamicin SA; ebselen; ifosfamide; edelfosine;Eculizumab; Eflornithine; Elemen; Emitefur; Epirubicin; Epristeride; Estramustine Analogues; Estrogen Agonists; Estrogen Antagonists; Etazolate; Etoposide Phosphate; Exemestane; Fadrozole; Fazarabine; Fenretinide; Filgrastim; Finasteride; Flavopiridol; Fludiazepam; Fluasterone; Fludarabine; Fluorouracil; Fostriecin; Fotemustine; Gadolinium Texaphyrin; Gallium Nitrate; Galocitabine; Ganirelix; Gelatinase Inhibitors; Gemcitabine; Glutathione Inhibitors; Heptatisulfan; Heregulin; Hexamethylenebisacetamide; Hypericin; Ibandronic Acid; Idarubicin; Idoxifene; Idramantone; Imophos; Ilomastat; Imatinib (such as Glivec); Imiquimod; Immunostimulatory Peptides; Insulin-like Growth Factor-1 Receptor Inhibitors; Interferon Agonists; Interferon; Interleukin; Iobenguane; Iodoxorubicin; 4-Isonicotinic Acid; Ipomperazole; Irsogladine; Isoxazolyl Benzene; Isoghomohalicondrin B; Itasert; Jasplakinolide; Kahalalide F; Lamellarin-N Triacetate; Lanreotide; Leinamycin; Lenograstim; Lentinan Sulfate; Leptolstatin; Letrozole; Leukemia Inhibitory Factor; Leukocyte Alpha Interferon; Leuprolide + Estrogen + Progesterone; Leuprorelin; Levamisole; Liarozole; Linear Polyamine Analogues; Lipophilic Disaccharide Peptides; Lipophilic Platinum Compounds; Lissoclinamide 7; Lobaplatin; Lombricine; Lomtriptorelin; Lonidamine; Losoxantrone; Losoxoribine; Letotecan; Lutetium Texaphyrin; Theophylline; Lytic Peptides; Maytansine; Mannostatin A; Marimastat; Masoprocol; Maspin; Matrix Metalloproteinase Inhibitors; Menogaril; Mebaral; Mitrelin; Methioninase; Metoclopramide; MIF Inhibitors; Mifepristone; Miltefosine; Mitobronitol; Mitomycin Analogues; Mitonafide; Mitogenic Fibroblast Growth Factor-Saponin; Mitoxantrone; Mofarotene; Moramustine; Erbitux, Human Chorionic Gonadotropin; Monophosphoryl Lipid A + Mycobacterium Cell Wall SK; Dipyridamole; Mustard Gas Anticancer Agents; Mycaperoxide B; Mycobacterium Cell Wall Extracts; Myriaporone; N-Acetyl Denarol; N-Substituted Benzamides; Nafarelin; Nartograstim; Naloxone + Pentazocine; Naproxen; Nartograstim; Nedaplatin; Nemorubicin; Nelidronic Acid; Nilutamide; Nishamycin; Nitric Oxide Modulators;Nitrogen oxide antioxidants; nitrilotris(methylenephosphonic acid); oblimersen(; O6-Benzylguanine; Octreotide; Okicenone; Oligonucleotide; Onapristone; Ondansetron; Ondansetron; Olaxin; Oral cytokine inducer; Ormaplatin; Oxaliplatin; Oxamycin; Paclitaxel; Paclitaxel analogues; Paclitaxel derivatives; Palauamine; Palmitoylrhizoxin; Pamidronic acid; Panaxytriol; Panomifene; Parabactin; Pazelliptine; Pegaspargase; Perdixine; Pentosan polysulfate sodium; Pentanoate; Tebuconazole; Pafuramidine; Peplomycin; Perillyl alcohol; Phenazineomycin; Phenylacetic acid; Phosphatase inhibitor; Picibanil; Pilocarpine hydrochloride; Pirarubicin; Picotamide; Platting A; Placental B; Plasminogen activator inhibitor; Platinum complex; Platinum compound; Platinum triamine complex; Porfimer sodium; Porphinomycin; Prednisone; Propyl bisacridone; Prostaglandin J2; Proteasome inhibitor; Protein A-based immunomodulator; Protein kinase C inhibitor; Protein kinase C inhibitor, microalgae; Protein tyrosine phosphatase inhibitor; Purine nucleoside phosphorylase inhibitor; Rhodoporphyrin; Pyrazoloacridine; Pyridoxylated hemoglobin polyoxyethylene conjugate; Raf antagonist; Raltitrexed; Ramosetron; Ras farnesyl protein transferase inhibitor; Ras inhibitor; Ras-GAP inhibitor; Demethylretifolin; Rhenium Re 186 hydroxyethylidene diphosphonate; Rhizoxin; Ribozyme; RII retinoic acid; Rofecoxib; Romurtide; Roquinimex; Rubiginone B1; Ruboxyl; Saffron alcohol; Santopine; SarCNU; Sarcophytol A; Sargramostim; Sdi 1 analogue; Semustine; Senescence-derived inhibitor 1; Sense oligonucleotide; Signal transduction inhibitor; Sizofiran; Sobuzoxane; Sodium borate; Sodium phenylacetate; Solvelol; Somatomedin-binding protein; Sonermin; Sparfosic acid; Spicamycin D; Spiroxatrine; Splenin; Spongistatin 1; Squalamine; Stemregenin 1; Matrix metalloproteinase inhibitor; Sulfinositol; Superactive vasoactive intestinal peptide antagonist; Suradista; Suramin; Swainsonine; Tamoxifen; Methyl iodide tamoxifen; Tauromustine; Tazarotene; Ticagrelan sodium; Tegafur; Telluropyran; Telomerase inhibitor; Temoporfin; Teniposide; Tetrachloro decoxide; Tetrazamine; Solanine; Thiocolchicoside; Thrombopoietin; Thrombopoietin analogue; Thymalfasin; Thymopoietin receptor agonist; Thymine; Thyroid-stimulating hormone; Tin ethyl etiopurpurin; Tirapazamine; Titanium dichloride; Topsentin; Toremifene; Translation inhibitor; Retinoic acid; Triacetyluridine; Trabectedin; Trimetrexate; Triptorelin; Tropisetron; Tolusterone; Tyrosine kinase inhibitor; Tyrosine phosphorylation inhibitor; UBC inhibitor; Ubenimex; Urogenital sinus-derived growth inhibitory factor; Urokinase receptor antagonist; Vapreotide; Varelin B; Veraguensin; Veramine; Verdine; Verteporfin; Vinorelbine;Changchun Sartin; Vitamin A; Vorozole; Zanoterone; Zinid Platinum; Zilacob; and Zinostatin Stimalamer.

[0235] In one embodiment, the additional therapeutic agent is a steroid. Those of ordinary skill in the art will understand that steroids have various medical uses, including but not limited to: (1) anti-inflammatory uses, such as betamethasone, budesonide, cortisone, dexamethasone, hydrocortisone, methylprednisolone, prednisolone, prednisone, and triamcinolone; (2) antiemetic uses, such as dexamethasone, hydrocortisone, and prednisone; (3) diagnostic uses, such as dexamethasone, for detecting Cushing's syndrome; and (4) immunosuppressant uses, such as betamethasone, cortisone, dexamethasone, hydrocortisone, methylprednisolone, prednisolone, prednisone, and triamcinolone. In addition, those of ordinary skill in the art can understand that corticosteroid drugs can be used as components in ophthalmic products (to treat various eye diseases), inhalers (to treat asthma or bronchial diseases), nasal drops and sprays (to treat various nasal diseases), and topical products such as ointments and creams (to treat various skin diseases).

[0236] Although various specific embodiments / aspects have been shown and described, it should be understood that various changes can be made without departing from the spirit and scope of the present disclosure.

[0237] All publications, patents, patent applications, and / or other documents cited in this application are hereby incorporated by reference in their entirety to the extent that each publication, patent, patent application, and / or other document cited in this application is separately stated to be incorporated by reference for all purposes.

[0238] Examples

[0239] Repurposing of Antibodies

[0240] The repurposing of antibodies can be used for a variety of applications. In one example, FDA-approved antibodies can be used to safely clear biothreat pathogens using the human immune system. This approach has many advantages:

[0241] Utilize highly optimized, safe, and low-cost antibodies to accelerate drug development;

[0242] Able to rapidly generate Technein repurposing agents, which can shorten the response time to newly infected pathogens: <6 months instead of years; platform technology with multiple product outputs. Repurposing drugs approved by health regulatory agencies (such as the FDA) may shorten the approval time.

[0243] Example 1

[0244] New Identification Technology for SARS-CoV-2 Spike Protein

[0245] Techneins that bind to the SARS-CoV-2 spike protein (i.e., the second target) were screened from the DHQ10 library. Several Techneins were identified. Figures 2A - 2C These Techneins contain amino acid and non-amino acid building blocks. Figure 2D 。

[0246] Using the spike protein RBD domain as the target, the binding affinity of each Technein was determined by microscale thermophoresis (MST). Some results are summarized in Table 4. The MST experiments were performed on a Monolith NT.115 pico (NanoTemper Technologies GmbH, Munich, Germany). The measurements were carried out at room temperature, repeated three times, with incubation periods of 15, 30, and 45 minutes respectively. The binding affinity was obtained from a 16-point, two-fold dilution series, with the ligand starting concentration of 1 μM and the target concentration of 5 nM. The SRBD protein was labeled using the Nanotemper Monolith second-generation protein labeling kit (maleimide-647-dye). The buffer for SRBD contained 20 mM HEPES pH 7.4, 150 mM NaCl, 10 mM MgCl2, and 0.05% Tween-20. The triplicate data were analyzed using MO.AffinityAnalysis software (NanoTemper Technologies GmbH).

[0247] Table 4: Binding affinities of exemplary Techneins

[0248] Hit ID Target KD (nM) 16838-89-1 SRBD - wt 36 16838-89-2 SRBD - wt 18 16838-89-3 SRBD - wt 240 16838-89-4 SRBD - wt 123 16838-89-5 SRBD - wt 70 16838-89-6 SRBD - wt 210 16838-89-7 SRBD - wt 85 16838-89-8 SRBD - wt 96 16838-89-9 SRBD - wt 199 16838-89-10 SRBD - wt 92 16838-89-11 SRBD - wt 310 16838-89-12 SRBD - wt No binding 16838-89-13 SRBD - wt 235 16838-89-14 SRBD - wt 216 16838-89-15 SRBD - wt 200 16838-89-16 SRBD - wt 119 16838-89-17 SRBD - wt 185 16838-89-18 SRBD - wt No binding 16838-89-19 SRBD - wt No binding 16838-89-20 SRBD - wt No binding

[0249] The Technein subgroup was identified as a lead hit and tested against different spike proteins in Biolayer Interferometry (BLI). The results are summarized in Table 5. BLI analysis was performed using a Sartorius Octet instrument. In a buffer (20 mM MOPS, 25 mM KCl, 0.02% Tween-20), Techneins in the concentration range of 1 - 250 nM were captured using a SAX biosensor dip and read for 600 seconds. The loaded sensor was quenched for 250 seconds with 25 μg / mL biotin (MOPS). Then, the baseline was measured in the case of MOPS buffer only. The sensor was then dipped into a 3-fold dilution series of spike RBD analyte (MOPS buffer) from 250 nM to 1.029 nM for 600 seconds. The sensor was then dipped into the buffer for 800 seconds for dissociation. The data was analyzed using HT11.1 analysis software and Savitzky-Golay filtering was used to reduce high-frequency noise. A global 1:2 fit was performed to determine the association rate constant (ka), dissociation rate constant (kd), and kinetic affinity (KD).

[0250] Table 5: Binding Affinities of Selected Techneins

[0251]

[0252] Example 2

[0253] Identification of New TECHNEINs Against Anti-HERCEPTIN Antibody

[0254] The DAAP2 and DHQ11 libraries were screened to find Techneins that bind to the Herceptin antibody. Many hits were identified. Figure 3A and 3B . These Techneins also contain amino acid and non-amino acid monomers. Figure 3C and 3D .

[0255] The binding affinity to Herceptin was tested in BLI. The results are summarized in Table 6.

[0256] Table 6: Binding Affinities of Selected Techneins Against Herceptin

[0257] Hit ID KD (nM) 16989-11-17 6 16910-59-7 150 16989-3-2 472 16989-3-1 477 16989-3-14 483 16989-3-13 559 16910-59-5 653 16989-3-6 730 16989-3-17 1340 16989-3-19 1360 16989-3-9 1360 16910-59-6 4650

[0258] Example 3

[0259] Repurposing Agents Against Spike Proteins

[0260] The anti-spike protein Technein (second molecule) is conjugated to biotin (first molecule) via a linker added to its C-terminus. Figure 4 As shown below, these Techneins can be used to repurpose anti-biotin antibodies (first target) to serve as anti-spike protein antibody-Technein conjugates (second target). Figure 7 .

[0261] Anti-spike Technein (second molecule) is conjugated to α-Gal (also known as alpha-Gal) (first target and also first molecule) in the presence or absence of a dye ( Figure 5A ) via a linker that includes PEG attached to the Cys side chain. Figure 5B ). Other anti-spike Technein-α-Gal conjugates are described in Figure 5C and 5D . As shown below, this synthetic compound is used to repurpose endogenous anti-α-Gal (first target) antibodies against anti-spike protein (second target). Figure 10 .

[0262] Anti-spike protein Technein (second molecule) is conjugated to anti-Herceptin Technein (first molecule) via a linker in the presence ( Figure 6A ) or absence ( Figure 6B ) of a dye. As shown below, this synthetic compound can repurpose anti-Herceptin antibodies (trastuzumab) to serve as anti-spike protein antibodies. Figure 9 .

[0263] Example 4

[0264] Analysis of antigen (SARS-COV2 SRBD) bead binding phagocytosis (antibody-dependent cellular phagocytosis, ADCP) assay by flow cytometry

[0265] Preparation of SARS-CoV-2 SRBD-coated beads by antigen adsorption. A total of 130 million carboxylated latex fluorescent beads with a diameter of 1 μm were incubated overnight at 4 °C with protein at a concentration of 40 μg / ml in 1 ml of PBS. Subsequently, the beads were washed twice with PBS plus 1% BSA and resuspended in RPMI medium. Three different biotinylated anti-SRBD Techneins were pre-incubated with mouse anti-biotin antibody or mock medium at 37 °C for 30 minutes to form a complex. Then the Technein-Ab complex (recycled anti-biotin antibody against SRBD) was added to the prepared SRBD beads and incubated at 37 °C for 1 hour. Then Raw264.7 cells were co-incubated with Technein / anti-biotin antibody-coated beads at 37 °C for 1 hour for phagocytosis. Subsequently, the cells were stained extracellularly with rabbit anti-SRBD Ab and goat anti-rabbit AF647. The phagocytosis efficiency (Y-axis in the figure) was calculated by the percentage of Raw264.7 cells with beads and lacking extracellular anti-rabbit AF647 staining. (Technein 77-4: 16877-77-4; 34-3: 16877-34-3, 76-2: 16788-76-2). Data are represented as mean ± SEM (n = 3). ADCP was promoted by the recycling agent. Figure 8 。

[0266] Example 5

[0267] In a sandwich enzyme-linked immunosorbent assay (ELISA), antibody recycling TECHNEINS bind to anti-α-GAL antibody and SARS-CoV2 antigen (SARS-CoV2 SRBD or Omicron S1 / S2 ECD).

[0268] Coat ELISA microplates with SARS-Cov2 SRBD or Omicron S1 / S2 ECD, incubate overnight at 4°C with 50 μl / well of PBS, with a protein concentration of 10 μg / ml. Subsequently, wash the microplates four times with PBS plus 0.05% Tween20 (PBST), and then block the reaction with PBST plus 1% BSA for 2 hours. Add the recycled SARS-CoV-binding Technein (89-8-aGal) and the uncoupled control SARS-CoV-binding Technein (89-8) at the indicated concentrations to 50 μl / well of 10 mM HEPES / 1% DMSO / 0.05% Tween 20 and incubate for 1 hour at room temperature. After washing, add 50 μl / well of mouse anti-α-Gal antibody (1:1000, m86, Absolute Antibody) and further incubate for 1 hour at room temperature. ELISA signals are developed using horseradish peroxidase (HRP)-conjugated anti-mouse IgG and 3,3',5,5'-tetramethylbenzidine (TMB) substrate. (Blank: negative control without any coated antigen; secondary antibody only: negative control without recycled Technein or control Technein). The results are shown in Figure 11 。The recycling agent αGal-89-8 exhibits antibody recycling of αGal antibodies, enabling them to bind to spike proteins, including wild-type SRBD and full-spike Omicron. As a negative control, compound 89-8 without α-Gal was used, showing a significantly lower level of antibody recycling.

[0269] Example 6

[0270] ADCP assay using HERCEPTIN antibody recycling agent

[0271] Α-GAL-TECHNEIN 89-8.

[0272] Wash 2.8-μm streptavidin-labeled magnetic Dynabeads TM M-280 (ThermoFisher) in PBS containing 2% BSA (PBS2%BSA) and incubate with 0.2 μg of biotinylated HER2 ( Figure 12A ) or biotinylated SRBD ( Figure 12B ) or biotinylated BSA ( Figure 12B) Incubate together, each mixed with 0.2 μg / ml biotin-FTIC in 250 μl PBS 2% BSA overnight at 4 °C. Wash the beads in PBS 2% BSA to remove unbound proteins and dyes, then resuspend them at a 1:100 dilution in RPMI 1640 containing 10% FCS. Then:

[0273] (A) Incubate 20 μl of biotinylated HER2-coated beads with 0.2 μg / ml HER2-specific antibody Herceptin in RPMI 1640 containing 10% FCS at 37 °C for 30 minutes in triplicate wells. As a negative control, simultaneously incubate another 20 μl of biotinylated HER2-coated beads with 0.2 μg / ml irrelevant human hIgG in RPMI 1640 containing 10% FCS at 37 °C for 30 minutes in triplicate wells. Figure 12A .

[0274] (B) Pre-incubate the recycling agent α-Gal-89-8 (200 nM) with 1:10 diluted human serum (= human serum preparation) or mouse serum (= mouse serum preparation) at room temperature for 1 hour (unlike human serum, mouse serum lacks αGal antibodies). Incubate 20 μl of biotinylated-SRBD-coated beads with the human serum preparation in RPMI 1640 containing 10% FCS at 37 °C for 30 minutes in triplicate wells. As a negative control, simultaneously treat 20 μl of biotinylated-BSA-coated beads in the same manner as the biotinylated-SRBD-coated beads, i.e., incubate them with the human serum preparation in RPMI 1640 containing 10% FCS at 37 °C for 30 minutes in triplicate wells. Set up two additional negative controls in parallel, using 20 μl of biotinylated SRBD-coated beads and 20 μl of biotinylated BSA-coated beads, each incubated with the mouse serum preparation in RPMI 1640 containing 10% FCS at 37 °C for 30 minutes in triplicate wells. Figure 12B .

[0275] (A + B) Then add 4 × 10^4 effector monocytes THP-1 to all opsonized beads. Spin the plate at 300 g for 1 minute to promote contact, then further incubate at 37 °C for 3 hours. Fix the cells with 4% paraformaldehyde and immediately acquire them on an A3 Symphony flow cytometer (BD Biosciences). The total phagocytosis score is determined by multiplying the percentage of cells with beads by their MFI. Figure 12A , B.

[0276] (C) To calculate the antigen-specific phagocytosis score of biotinylated HER2-coated beads shown in A, the total phagocytosis score obtained in the presence of the irrelevant hIgG was subtracted from the phagocytosis score obtained in the presence of the HER-2 specific antibody Herceptin. Figure 12C 。

[0277] (C) To calculate the antigen-specific phagocytosis scores of biotinylated SRBD-coated beads and biotinylated BSA-coated beads shown in B, the total phagocytosis score obtained in the presence of a human serum preparation was subtracted from the total phagocytosis score obtained in the presence of a mouse serum preparation. Figure 12A -C.

[0278] Example 7

[0279] Binding of SARS-COV-2 Spike Protein to Technein Monomers and Dimers

[0280] The binding of Technein to SARS-CoV-2 SRBD (wild type) was demonstrated by ELISA assay. The recombinant SRBD protein (Sino Biological) was immobilized on a 96-well ELISA plate containing 1% BSA, 0.1% Tween-20 in PBS, pH 7.4. The wells were washed thoroughly with buffer, and then biotinylated Technein 76-3 monomer (100 nM) and biotinylated Technein dimer 16877-9-4 (100 nM) were applied to the wells, with a background control without Technein. After washing, the wells were treated with streptavidin-HRP to measure the degree of binding, and the absorbance was read at 405 nm. Figure 13.

[0281] Example 8

[0282] Method for Rapid Vaccination by Antibody Repurposing

[0283] Antibody repurposing can generate rapid immunity to a pathogen by first administering a therapeutic antibody that binds to a known antigen or hapten. For example, the FDA-approved antibody drug trastuzumab (Herceptin), which was developed to bind to the protein HER2. Administering a bifunctional repurposing agent (synthetic compounds of the present disclosure) that can bind to the antibody through a first molecule and to a second target on a viral pathogen such as the SARS-CoV-2 spike protein through a second molecule can target the original antibody to the pathogen and generate an immune response to clear the pathogen from circulation. This approach enables a common antibody component to be used with a variety of different repurposing agents that are specific for various types of pathogens that cause human disease. Figure 14 。

Claims

1. A synthetic compound comprising at least one unit, each unit comprising: (i) a first molecule capable of binding to an antibody or a target-binding fragment thereof; wherein the antibody is capable of binding to a first target; and (ii) a second molecule capable of binding to a second target; wherein the first target and the second target are different, and wherein the first molecule and the second molecule are directly covalently linked or covalently linked via a linker.

2. The synthetic compound according to claim 1, wherein the first molecule comprises the first target, an epitope thereof, an antibody-binding fragment thereof or a first synthetic polymer; and the second molecule comprises a second synthetic polymer capable of binding to the second target, wherein the first and second synthetic polymers comprise amino acid monomers and / or non-amino acid monomers.

3. The synthetic compound according to claim 1, wherein the first molecule non-covalently binds to one or more complementarity-determining regions (CDRs) of the antibody.

4. The synthetic compound according to claim 1, wherein the antibody has been approved by one or more agencies selected from the US Food and Drug Administration (FDA), the European Medicines Agency (EMA), the Swiss Medicines Agency, the National Medical Products Administration (NMPA) and the Japanese Pharmaceuticals and Medical Devices Agency (PMDA).

5. The synthetic compound according to claim 4, wherein the antibody is selected from trastuzumab (Herceptin), adalimumab (Humira), bevacizumab (Avastin), rituximab (Rituxan / MabThera), infliximab (Remicade) or any other antibody in Table 1.

6. The synthetic compound according to claim 1, wherein the antibody is an endogenous antibody.

7. The synthetic compound according to claim 6, wherein both the first target and the first molecule are α-Gal.

8. The synthetic compound according to claim 1, wherein the target is selected from proteins, sugars, carbohydrates, nucleic acids, lipids and combinations thereof.

9. The synthetic compound according to claim 1, wherein the target is a viral target, a tumor-specific target (e.g., a tumor-associated antigen), a tissue-specific target, a cell-specific target, a bacterial target, a fungal target and combinations thereof.

10. The synthetic compound according to claim 1, wherein the first target is human epidermal growth factor receptor 2 (HER2) and / or the second target is a viral protein (including but not limited to the SARS-CoV-2 spike protein).

11. The synthetic compound according to claim 2, wherein the amino acids include L-amino acids, D-amino acids, β-amino acids, γ-amino acids or combinations thereof, and the non-amino acid monomers include PAM, DhqF, DhqB, DhqO, DhqY, DhqE, N-substituted glycines, triazines, pyrimidines or combinations thereof.

12. The synthetic compound according to claim 1, wherein the linker comprises polyethylene glycol (PEG), a polyglycine sequence, a peptide, an alkyl chain, cysteine, lysine, glutamine, maleimide, dibenzyloctane, or a combination thereof.

13. The synthetic compound according to claim 1, wherein at least one unit comprises a plurality of first and / or second molecules, wherein: i. the plurality of first molecules are directly covalently bound to each other or covalently bound to each other through a linker; and / or ii. the plurality of second molecules are directly covalently bound to each other or covalently bound to each other through a linker; Preferably, wherein the linker comprises PEG, Lys, Gln, or a combination thereof.

14. The synthetic compound according to claim 1, wherein the first molecule and / or the second molecule binds to the antibody or the second target, respectively, with an affinity lower than 500 nM, preferably lower than 200 nM, more preferably lower than 100 nM.

15. The synthetic compound according to claim 1, wherein the first molecule and / or the second molecule do not significantly bind to any other target, and are also referred to as being specific for the first target or the second target, respectively.

16. A conjugate comprising an antibody or an antigen-binding fragment thereof non-covalently bound to at least one of the synthetic compounds according to any one of claims 1 to 15, wherein the conjugate is also referred to as a repurposed antibody or a repurposed fragment thereof.

17. The conjugate according to claim 16, wherein the antibody comprises light and heavy chain immunoglobulin variable and constant regions.

18. The conjugate according to claim 16, wherein the antibody or its antigen-binding fragment is selected from: monoclonal antibodies, recombinantly produced antibodies, monospecific antibodies, multispecific antibodies (including bispecific antibodies), human antibodies, engineered antibodies, humanized antibodies, chimeric antibodies, immunoglobulins, synthetic antibodies, tetrameric antibodies comprising two heavy chain and two light chain molecules, antibody light chain monomers, antibody heavy chain monomers, antibody light chain dimers, antibody heavy chain dimers, antibody light chain - antibody heavy chain pairs, intracellular antibodies, antibody fusions (sometimes referred to herein as "antibody conjugates"), hetero-conjugate antibodies, single domain antibodies, monovalent antibodies, single chain antibodies or single chain Fv (scFv), camelized antibodies, affibodies, Fab, Fab', F(ab')2 and Fv fragments, disulfide-linked Fv (sdFv), anti-idiotypic (anti-Id) antibodies (including, for example, anti-anti-Id antibodies), minibodies, domain antibodies, synthetic antibodies (sometimes referred to herein as "antibody mimetics"), and antigen-binding fragments of any of the above antibodies.

19. The conjugate according to any one of claims 16 to 18 and / or the synthetic compound according to any one of claims 1 to 15, wherein the antibody that binds to the first molecule, the conjugate (repurposed antibody), and / or the synthetic compound is an immunotherapeutic agent.

20. A method of repurposing an antibody or a first target-binding fragment thereof that binds to a first target by changing the target of the antibody or the first target-binding fragment from the first target to a second target, the method comprising non-covalently coupling the antibody or the first target-binding fragment thereof to at least one synthetic compound according to any one of claims 1 to 15, wherein the second target is also referred to as the repurposed target, and wherein the conjugate is also referred to as the repurposed antibody.

21. A method of treating a disorder in a subject in need thereof with a repurposed antibody, which comprises administering to the subject an effective amount of the synthetic compound (or a portion thereof) of claims 1 to 15, the antibody according to any one of claims 1 to 15, and / or the repurposed antibody according to any one of claims 16 to 19.

22. The method according to claim 21, wherein the disorder is an infectious disease caused by a pathogen comprising the second target or cancer expressing the second target.

23. The method according to claim 22, wherein the pathogen is a virus (including but not limited to the SARS-CoV-2 coronavirus).

24. The method according to any one of claims 20 to 23, wherein the antibody is an endogenous antibody.

25. A method for treating a disease caused by a pathogen in a subject in need thereof, which comprises: (i) administering to the subject an antibody that binds to a first molecule of a synthetic compound according to any one of claims 1 to 15, or confirming that the subject carries an endogenous antibody that binds to the compound; (ii) exposing the subject to or the subject having been exposed to a pathogen, an epitope of the pathogen, and / or a portion of the pathogen; and (iii) administering to the subject the synthetic compound of (i), wherein the pathogen comprises the second target of the synthetic compound.

26. The method according to claim 25, wherein the pathogen causes a disease selected from viral diseases, bacterial diseases, fungal diseases, or tick-related diseases.

27. A method of ameliorating, treating, or reducing the incidence of cancer / tumor / malignancy in a subject in need thereof, preferably, wherein the method aids or enhances the immune system to kill or eradicate cancer cells, and wherein the method comprises administering to the subject a therapeutically effective amount of one or more conjugates / repurposed antibodies, synthetic compounds, or antibodies according to any one of claims 1 to 19, preferably an antibody that initiates an active (or achieves a passive) immune response to destroy cancer cells.

28. A method of ameliorating, treating, or reducing the incidence of cancer / tumor / malignancy in a subject in need thereof, preferably, wherein the method aids or enhances the immune system to kill or eradicate cancer cells, and wherein the method comprises administering to the subject a therapeutically effective amount of CAR-T cells, T cells, and / or tumor-infiltrating lymphocytes that have contacted or bound to an immunotherapeutic compound according to any one of claims 1 to 19, preferably, wherein the cells initiate an active (or achieve a passive) immune response to destroy cancer cells.

29. A method of using a therapeutically effective amount of a conjugate / repurposed antibody, synthetic compound, or antibody of any one of claims 1 to 19, or an immunotherapeutic cell conjugated or contacted with a compound or conjugate of any one of claims 1 to 19 (including the use of autologous and / or allogeneic cells or immortalized cell lines) in vivo or ex vivo as adoptive immunotherapy for treating cancer in a subject in need thereof, which comprises administering or giving to the subject one or more conjugates / repurposed antibodies, synthetic compounds, antibodies of any one of claims 1 to 19 and / or contacting the immunotherapeutic cells with one or more conjugates / repurposed antibodies, synthetic compounds, antibody cells and / or nucleic acids of any one of claims 1 to 19.

30. A composition comprising one or more conjugates / repurposed antibodies, synthetic compounds, and / or antibodies of any one of claims 1 to 19, and / or CAR-T cells, T cells, or TILs conjugated or contacted with a compound of any one of claims 1 to 19, preferably wherein the composition is a pharmaceutical composition.

31. An immunotherapeutic compound of any one of claims 1 to 19 or the composition of claim 30 for cancer immunotherapy, wherein the immunotherapeutic compound or composition comprises a repurposed antibody, fragments thereof, and / or synthetic compounds of the present disclosure, and / or CAR-T cells, T cells, and / or tumor-infiltrating lymphocytes conjugated or contacted with a compound of any one of claims 1 to 19.

32. Use of a synthetic compound according to any one of claims 1 to 15, a conjugate according to any one of claims 16 to 19, a composition comprising the same, and / or CAR-T cells, T cells, or TILs conjugated or contacted with a compound according to any one of claims 1 to 19 in the treatment of a disease or disorder.

33. The use according to claim 32, wherein the disease or disorder is an infectious disease or disorder or cancer, or any other disease disorder described in the specification.

34. Use of a synthetic compound according to any one of claims 1 to 15, a conjugate according to any one of claims 16 to 19, a composition comprising the same, and / or CAR-T cells, T cells, or TILs conjugated or contacted with a compound according to any one of claims 1 to 19 in the manufacture of a medicament for treating a disease or disorder, preferably wherein the disease or disorder is an infectious disease or disorder or cancer, or any other disease disorder described in the specification.

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