KR20230128271A - HER3 radioimmunotherapy for the treatment of solid cancers - Google Patents

Abstract

radionuclides such as ²²⁵ Ac; in a subject by administering an effective amount of a HER3-targeting agent labeled with ¹⁷⁷ Lu, ¹³¹ I, ⁹⁰ Y, ²¹³ Bi, ²¹¹ At, ²¹³ Bi, ²²⁷ Th, or ²¹² Pb, alone or in combination with another therapeutic agent or treatment modality. Compositions and methods for treating solid cancers, such as HER3-positive tumors, are provided. An effective amount of the radiolabeled HER3-targeting agent can be the maximum tolerated dose administered as a single bolus or together in divided doses equal to the maximum tolerated dose.

Description

HER3 radioimmunotherapy for the treatment of solid cancers

CROSS REFERENCES TO RELATED APPLICATIONS

This application claims International Application No. PCT/US21/56259, filed on October 22, 2021, and its priority applicant, U.S. Provisional Application No. 63/250,725, filed on September 30, 2021, and filed on July 28, 2021. US Provisional Application No. 63/226,699; US Provisional Application No. 63/118,181, filed on November 25, 2020; and U.S. Provisional Application No. 63/116,225, filed on November 20, 2020, each of which is incorporated herein by reference in its entirety.

sequence listing

This application contains a sequence listing submitted electronically in ASCII format, the entire contents of which are incorporated herein by reference. Said ASCII copy, created on November 22, 2021, is named ATNM-010PCT_SL_ST25.txt and is 191,107 bytes in size.

field of invention

The present invention relates to the field of radiation therapy.

ErbB3/HER3 is a type I transmembrane glycoprotein that is a member of the erythroblast oncogene B (ErbB) family of tyrosine kinase receptors (EGFR, HER2, HER3 and HER4). Signaling through HER3 can be activated in a ligand-dependent or ligand-independent manner. In the absence of ligand, the HER3 receptor molecule is normally expressed on the cell surface as a monomer with a conformation that prevents receptor dimerization. In this conformation, the dimerization loop of subdomain II makes intramolecular contact with a pocket on subdomain IV. Binding of HER3 ligands such as neuregulin (NRG), eg NRG1 (also known as heregulin, HRG) or NRG2, to subdomains I and III of the extracellular domain results in exposure of the dimerization loop of subdomain II. resulting in conformational changes that facilitate receptor dimerization and signaling. Certain cancer-associated mutations in HER3 disrupt this interaction between subdomains II and IV, an interaction required for the formation of an inactive 'closed' conformation, thus constitutive presentation of a dimerization loop in the absence of ligand binding. and activation of HER3-mediated signaling.

Antibodies that target HER3 can be used to target specific cancer cells, particularly certain solid cancers. HER3 is overexpressed in several types of cancer, such as breast, gastrointestinal and pancreatic cancer. A correlation has been shown between the expression of HER2/HER3 and the progression of these cancers from non-invasive to invasive stages. Agents that interfere with HER3-mediated signaling, such as anti-HER3 antibodies, can allow the establishment of robust immune responses against cancer cells that may be inappropriate using conventional therapies.

Accordingly, one object of the invention disclosed herein is to provide therapeutically effective radiolabeled HER3 targeting agents, such as for the treatment of HER3-positive cancers. A related object of the invention disclosed herein is to provide a method of treatment comprising administering such a radiolabeled HER3 targeting agent, alone or in combination with one or more additional therapeutic agents.

The present invention uses a radioisotope-labeled, HER3-targeting agent that targets HER3, such as a monoclonal antibody, peptide, or small molecule, and a radiolabeled HER3-targeting agent to treat HER3-positive (HER3-expressing) cancers. Methods of diagnosis and/or treatment are provided.

According to certain aspects of the invention, radiolabeled HER3 targeting agents useful for diagnostic purposes are anti-HER3 antibodies, peptides, or anti-HER3 antibodies comprising radioisotopes such as ¹¹¹ In, ⁶⁸ Ga, ⁶⁴ Cu, ⁸⁹ Zr, or ¹⁷⁷ Lu. It may be a small molecule.

According to certain other aspects, the radiolabeled HER3 targeting agent useful for therapeutic intervention is a radioisotope such as ¹³¹ I, ¹²⁵ I, ¹²³ I, ⁹⁰ Y, ¹⁷⁷ Lu, ¹⁸⁶ Re, ¹⁸⁸ Re, ⁸⁹ Sr, ¹⁵³ Sm. , ³² P, ²²⁵ Ac, ²¹³ Bi, ²¹³ Po, ²¹¹ At, ²¹² Bi, ²¹³ Bi, ²²³ Ra, ²²⁷ Th, ¹⁴⁹ Tb, ¹³⁷ Cs, ²¹² Pb, or a combination thereof; or a small molecule. According to certain preferred aspects, the radiolabeled HER3 targeting agent may comprise ¹³¹ I, ⁹⁰ Y, ¹⁷⁷ Lu, ²²⁵ Ac, ²¹³ Bi, ²¹¹ At, ²²⁷ Th, or ²¹² Pb.

According to certain aspects, the HER3-positive cancer may be a solid tumor.

The treatment methods of the invention disclosed by this application generally comprise administering to a patient a therapeutically effective amount of a radiolabeled HER3 targeting agent. According to certain aspects, the effective amount of the radiolabeled HER3 targeting agent can be a maximum tolerated dose (MTD) or can be divided doses, wherein the total amount of radiation administered in divided doses is the MTD.

According to certain aspects, a composition or amount of a HER3 targeting agent comprising a radiolabeled fraction and a non-radiolabeled fraction of the HER3 targeting agent is provided and/or used. As such, an effective amount of a HER3 targeting agent may include a total protein dose of less than 100 mg, such as 5 mg to 60 mg, or 5 mg to 45 mg. According to certain aspects, the total protein dose may be between 0.001 mg/kg and 3 mg/kg of the subject's body weight, such as between 0.005 mg/kg and 2 mg/kg of the subject's body weight. According to certain aspects, the total protein dose may be less than 2 mg/kg, or less than 1 mg/kg, less than 0.5 mg/kg, or even less than 0.1 mg/kg. A portion of the total protein dose is radiolabeled (ie, radioconjugated) as indicated, wherein the effective amount of the radiolabeled HER3 targeting agent may vary depending on the specific radioisotope selected. Preferred radioisotopes for therapeutic intervention include ²²⁵ Ac, ¹⁷⁷ Lu, ¹³¹ I, ⁹⁰ Y, ²¹³ Bi, ²¹¹ At, ²²⁷ Th, or ²¹² Pb. Thus, the HER3 targeting agent may include a radiolabeled fraction and an unlabeled fraction.

According to certain aspects, an effective amount of a HER3 targeting agent, i.e., a radiolabeled portion thereof, such as ^{a 225} Ac-anti-HER3 antibody, peptide, or small molecule, in terms of radiation dose, is a dose of 0.1 to 20 μCi/kg of subject's body weight. , such as a dose of 0.1 to 10 μCi/kg or 0.1 to 5 uCi/kg (body weight of the subject), or a dose of 0.5 to 20 μCi/kg or 1 to 10 uCi/kg (body weight of the subject).

According to certain aspects, an effective amount of a HER3 targeting agent, i.e., a radiolabeled portion thereof, such as ^{a 225} Ac-anti-HER3 antibody, peptide, or small molecule, is a component of the targeting agent, i.e., a full-length antibody or antigen-binding antibody fragment (eg eg, Fab, Fab ₂ , minibodies, nanobodies, etc.), such as any of those disclosed herein. For example, if the HER3 targeting agent comprises a full-length antibody, ²²⁵ Ac-anti-HER3 antibody, the dose is less than 5 uCi/kg of the subject's body weight, such as between 0.1 and 5 uCi/kg of the subject's body weight. can be Alternatively, if the HER3 targeting agent comprises a fragment ²²⁵ Ac-anti-HER3 antibody, the dose may be greater than 5 uCi/kg of the subject's body weight, such as between 5 and 20 uCi/kg of the subject's body weight. .

According to certain aspects, the HER3 targeting agent is an anti-HER3 antibody, such as a monoclonal antibody or antigen-binding fragment thereof, such as an IgG or antigen-binding fragment thereof, such as patritumab from Daiichi Sankyo, Merrimack Pharma Serivantumab (MM-121) from Merrimack Pharmaceuticals, rumletuzumab from Roche, elgemtumab from Novartis, GSK2849330 from GlaxoSmithKline, Celdex Thera CDX-3379 from Celldex Therapeutics, EV20 and MP-RM-1 from MediPharma, ISU104 from Isu Abxis Co., Hummingbird Bioscience Pte. and/or by HMBD-001 (10D1F) from Regeneron Pharmaceuticals, REGN1400 from Regeneron Pharmaceuticals, and/or AV-203 from AVEO Oncology. According to certain aspects, the anti-HER3 antibody is selected from one or more of patritumab, ceribantumab, rumletuzumab, elgemtumab, AV-203, CDX-3379, or GSK2849330.

According to certain aspects, the HER3 targeting agent is selected from the group consisting of administration once every 7 days, 10 days, 12 days, 14 days, 20 days, 24 days, 28 days, 35 days, and 42 days throughout the treatment period. It may be administered according to an administration schedule, wherein the treatment period includes at least two administrations.

According to certain aspects, the HER3 targeting agent is administered in two doses on day 1 and day 5, day 6, day 7, day 8, day 9, or day 10 of the treatment period, or on the first day of the treatment period. It may be administered according to a dosing schedule comprising two doses on days 1 and 8.

According to certain aspects, the HER3 targeting agent can be administered as a single bolus or infusion in a single subject specific dose.

According to certain aspects, the method may further comprise administration of one or more additional therapeutic agents, such as chemotherapeutic agents, small molecule drugs, anti-inflammatory agents, immunosuppressive agents, immunomodulatory agents, antimyeloma agents, cytokines, or combinations thereof. Exemplary chemotherapeutic agents include at least radiosensitizers that can synergize with radiolabeled HER3 targeting agents, such as temozolomide, cisplatin and/or fluorouracil.

According to certain aspects, the method may further include administration of one or more immune checkpoint therapies. Exemplary immune checkpoint therapies include CTLA-4, PD-1, TIM-3, VISTA, BTLA, LAG-3, TIGIT, CD28, OX40, GITR, CD137, CD40, CD40L, CD27, HVEM, PD-L1, PD -L2, PD-L3, PD-L4, CD80, CD86, CD137-L, GITR-L, CD226, B7-H3, B7-H4, BTLA, TIGIT, GALS, KIR, 2B4, CD160, CGEN-15049 antibodies, or any combination thereof. According to certain aspects, the immune checkpoint therapy may include an antibody to an immune checkpoint protein selected from the group consisting of antibodies to PD-1, PD-L1, PD-L2, CTLA-4, CD137, and combinations thereof. there is.

According to certain aspects, an immune checkpoint therapy can be administered to a subject in an effective amount, such as a dose of 0.1 mg/kg to 50 mg/kg of patient body weight, such as 0.1-5 mg/kg, or 5-30 mg/kg. there is.

According to certain aspects, the method may further include administration of one or more DNA damage response inhibitors (DDRi). Exemplary DDRi agonists are one or more antibodies or small molecules that target poly(ADP-ribose) polymerase (ie, poly(ADP-ribose) polymerase inhibitors or PARPi). PARPi can include, for example, olaparib, niraparib, rucaparib, thalazoparib, or any combination thereof. According to certain aspects, PARPi can be given in a subject effective amount including 0.1 mg/day - 1200 mg/day, such as 0.100 mg/day - 600 mg/day, or 0.25 mg/day - 1 mg/day. Exemplary subject effective amounts are 0.1 mg, 0.25 mg, 0.5 mg, 0.75 mg, 1.0 mg, 100 mg, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 700 mg, 750 mg, 800 mg, 900 mg, and 1000 mg, administered orally once or twice daily.

Another exemplary DDRi includes mutated telangiectatic ataxia (ATM), mutated and Rad-3 related telangiectatic ataxia (ATR), or an inhibitor of Weel. Exemplary inhibitors of ATM include KU-55933, KU-59403, wortmannin, CP466722 and KU-60019. Exemplary inhibitors of ATR include at least Schizandrin B, NU6027, NVP-BEA235, VE-821, VE-822, AZ20 and AZD6738. Exemplary inhibitors of Weel include AZD-1775 (ie, Adavosertip).

According to certain aspects, the method may further comprise administration of one or more CD47 blockers. CD47 blockers include monoclonal antibodies, SIRPα-Fc fusion proteins or other molecules that prevent CD47 from binding to SIRPα or otherwise block or downregulate the immunosuppressive activity of CD47, such as magnolimab, remzoparlimab, AO Includes -176, AK117, IMC-002, IBI-188, IBI-322, BI 766063, ZL-1201, AXL148, RRx-001, ES004, SRF231, SHR-1603, TJC4, TTI-621, or TTI-622 can do. Exemplary effective doses for CD47 blockers include 0.05 to 5 mg/kg of patient body weight. CD47 blockers may also include agents that modulate the expression of CD47 and/or SIRPα, such as nucleic acid approaches such as antisense, RNAi or μRNA approaches. Exemplary CD47 blockers also include phosphorodiamidate morpholino oligomers (PMOs) that block translation of CD47, such as MBT-001.

According to certain aspects, the method may further include administration of a combination of additional therapeutic agents. Exemplary combinations are directed against at least one or more DDRi and/or one or more immune checkpoint therapies and/or one or more CD47 blockers and/or one or more chemotherapeutic agents and/or one or more small molecule anticancer drugs and/or different cancer-associated antigens. Including one or more targeting agents that have been derived.

According to certain aspects, the radiolabeled HER3 targeting agent and one or more additional therapeutic agents may be administered simultaneously or sequentially. When more than one additional therapeutic agent is administered, these therapeutic agents may be administered simultaneously or sequentially.

According to certain aspects of the invention, the radiolabeled HER3 targeting agent may be a multispecific targeting agent, such as a multispecific antibody or a bispecific antibody, wherein at least one portion recognizes HER3. Accordingly, the method may comprise administering to a subject an effective amount of a multispecific antibody, wherein the multispecific antibody comprises a first target recognition component that specifically binds an epitope of HER3, and a first target recognition component and and a second target recognition component that specifically binds an epitope of a different HER3 or that specifically binds an epitope of a different antigen, such as a different cancer-associated antigen. According to certain aspects, the HER3 targeting agent is a multispecific antibody directed against a first epitope of HER3 and at least a second epitope of HER3, or against HER3 and at least a second (different) antigen. Exemplary multispecific antibodies that can be radiolabeled for diagnostic and/or therapeutic use according to the present invention include bispecific antibodies to HER3/HER2, such as MM-111 from Merrimack Pharmaceuticals or Merus ( Merus) MCLA-128 from N.V.; or a bispecific antibody to IGF-1R/HER3 such as MM-141 from Merrimac Pharmaceuticals (ie istiratumab); or a bispecific antibody to EGFR/HER3, such as MEHD7945A from Roche (i.e. duligotumab) or a cetuximab-based bispecific or multispecific zybody from Zyngenia Inc. including any

According to certain aspects, there is provided a composition comprising a mixture of a HER3 targeting agent, such as an antibody to HER3, and one or more additional targeting agents, such as antibodies targeting/to one or more different cancer-associated antigens, wherein the HER3 One or more of the targeting agent and the other targeting agent(s) may be radiolabeled or non-radiolabeled in any combination. Exemplary antibody compositions comprising antibody mixtures include at least Sym013 from Symphogen with 6 monoclonal antibodies against EGFR (HER1), HER2 and HER3. In one aspect, one or more of the antibodies of Sym013 can be radiolabeled in any combination, eg, with at least a HER3 antibody and no antibodies to EGFR and HER2, or with one or more of these antibodies.

Additional features, advantages and aspects of the present invention may be described or made apparent in consideration of the following detailed description, drawings, if any, and claims. Moreover, it is to be understood that both the foregoing summary of the invention and the following detailed description are exemplary and are intended to provide further explanation without limiting the scope of the claimed invention.

1 provides amino acid sequences of N-terminal regions, complementarity determining regions, and variable regions of heavy and light chains of HER3 monoclonal antibodies that can be implemented in various aspects of the present invention.
2 provides amino acid sequences of full-length heavy and light chains, with or without leader sequences, of HER3 monoclonal antibodies that can be implemented in various aspects of the present invention.
FIG. 3 shows the ELISA assay binding characteristics of Ac225 labeled DOTA-conjugated anti-HER3 monoclonal antibodies compared to unmodified anti-HER3 antibodies and non-specific antibodies (IgG), indicating that such modifications are immunoreactive to HER3. proves that it has no real effect on
Figure 4 shows 225Ac-HER3-ARC, unmodified anti-HER3 mAb, non-specific antibody control (IgG) and BxPC-3 cells (human pancreatic adenocarcinoma) on HER3-positive NCI-H1975 cells (human lung adenocarcinoma, NSCLC) and BxPC-3 cells (human pancreatic adenocarcinoma). A graph showing the results of a flow cytometry assay examining the binding of the secondary antibody only control.
5 is a graph showing the in vitro cytotoxic effect of 225Ac-HER3-ARC on the HER3-positive cell line NCI-H1975 as a function of radiation dose.
6A is a graph showing that 225Ac-HER3-ARC upregulates cell surface calreticulin (CRT) in NCI-H1975 cells.
6B is a graph showing that 225Ac-HER3-ARC upregulates CD47 on NCI-H1975 cells.
7A is a graph showing the results of a phagocytosis assay demonstrating that the combination of 225Ac-HER3-ARC with an anti-CD47 blocking antibody enhanced phagocytosis of BxPC-3 cells compared to either treatment alone.
7B is a graph showing the results of a phagocytosis assay demonstrating that the combination of 225Ac-HER3-ARC with an anti-CD47 blocking antibody enhanced phagocytosis of NCI-H1975 cells compared to either treatment alone.
8 is a graph showing the effect of 225Ac-HER3-ARC in combination with an anti-CD47 blocking antibody on tumor growth at different radiation doses in a human tumor (NCI-H1975 cell) mouse xenograft model.
9 is a graph showing body weight over time for the subjects of the experiment described in FIG. 8 .
10 is a graph showing survival probability over time for subjects in the experimental group of the experiment described in FIG. 8 .
Figure 11 shows vehicle (control), CD47 blocking antibodies Magrolimab alone, 225Ac-Trastuzumab alone, and Magrolimab and 225Ac-Trastuzumab in an NGS mouse xenograft model using the HER2-positive SK-OV3 human ovarian cancer cell line. It is a graph showing the comparative effect of combinations of zumab on tumor growth.
Figure 12 : Tumor growth of vehicle (control), magnolimab alone, 177Lu-trastuzumab alone, and the combination of magnolimab and 177Lu-trastuzumab in an NGS mouse xenograft model using the SK-OV3 human ovarian cancer cell line. It is a graph showing the comparative effect on

In one aspect, the invention disclosed by the present application provides compositions and methods for treating cancers that express HER3, ie, HER3-positive cancers. This aspect generally relates to the administration of an effective amount of a radiolabeled HER3 targeting agent, such as a radiolabeled antibody, peptide, or small molecule targeted to HER3, alone or in one or more additional therapeutic agents, to a mammalian subject in need of treatment, such as a human patient. and/or administration in combination with a treatment modality/treatment.

Additional therapeutic agents and treatment modalities that may be used include, for example, at least one or more immune checkpoint therapies and/or one or more inhibitors of components of the DNA damage response pathway (i.e., DNA damage response inhibitors, DDRi, such as poly(ADP- ribose) polymerase, i.e. PARPi) and/or one or more CD47/SIRPα axis blockers and/or one or more chemotherapeutic agents such as radiosensitizers, and/or one or more small molecule oncology drugs such as tyrosine kinase inhibitors , and/or one or more targeting agents to different antigens.

The invention disclosed herein further provides methods for identifying, imaging and/or diagnosing HER3-positive cancer in a subject. The invention disclosed by the present application further provides methods of identifying, imaging and/or diagnosing a HER3-positive cancer in a subject, and then treating the subject according to any of the methods disclosed herein.

Definitions and Abbreviations

The singular form includes the plural referent unless the context clearly dictates otherwise. Thus, for example, reference to an antibody includes both a single antibody and a plurality of different antibodies.

As used in this specification and claims, the word "comprising" and the forms of the word "comprising" as well as the forms of the word "comprising" and "comprising" do not limit the inclusion of elements beyond those with which they are recited. . Additionally, throughout this disclosure various aspects or elements thereof are described in terms of “comprising” or “comprising”, but their corresponding counterparts described in terms of “consisting essentially of” or “consisting of”. Aspects or elements are similarly disclosed. For example, while certain aspects of the invention have been described in terms of a method that “comprises” or “comprises” administering a radiolabeled targeting agent, it instead “consists essentially of” administering a radiolabeled target. or corresponding methods listed as “consisting of” are also within the scope of this aspect and are disclosed by this disclosure.

The term "about", when used in this disclosure in conjunction with numerical designations or values, to describe, for example, temperature, time, amount, and concentration, including in the description of a particular range, a variance of ±10%. , and, within that larger variance, represents a variance of ±5% or ±1% of the numerical designation or value.

As used herein, “administering” in reference to a targeting agent, such as an antibody, antibody fragment, Fab fragment, or aptamer, means delivering the agent to the body of a subject via any known method suitable for antibody delivery. means that Specific modes of administration include, but are not limited to intravenous, transdermal, subcutaneous, intraperitoneal, intrathecal and intratumoral administration. Exemplary methods of administration for the antibody may be substantially as described in International Publication No. WO 2016/187514, incorporated herein by reference.

Also in the present invention, antibodies may be formulated using, for example, one or more routinely used pharmaceutically acceptable carriers and excipients. Such carriers and excipients are well known to those skilled in the art. For example, injectable drug delivery systems include solutions, suspensions, gels, microspheres, and polymeric injectables, and include excipients such as solubility-altering agents (e.g., ethanol, propylene glycol, and sucrose) and polymers (e.g., ethanol, propylene glycol, and sucrose). For example, polycapryllactone and PLGA) may be included.

As used herein, the term “antibody” refers to (a) an immunoglobulin molecule comprising two heavy chains and two light chains and recognizing an antigen; (b) polyclonal and monoclonal immunoglobulin molecules; (c) monovalent and bivalent fragments thereof, such as Fabs, di-Fabs, scFvs, diabodies, minibodies, and nanobodies (sdAbs); (d) naturally occurring and non-naturally occurring, such as fully synthetic antibodies, IgG-Fc-silencing and chimeras; and (e) bispecific and multispecific forms thereof. Immunoglobulin molecules may be derived from any of the commonly known classes including, but not limited to, IgA, secretory IgA, IgG and IgM. IgG subclasses are also well known to those skilled in the art and include, but are not limited to, human IgG1, IgG2, IgG3 and IgG4. The N-terminus of each chain defines a “variable region” of about 100 to 110 or more amino acids primarily responsible for antigen recognition. The terms variable light chain (VL) and variable heavy chain (VH) refer to these regions of the light and heavy chains, respectively. Antibodies may be human, humanized or non-human. Where a specific aspect of the invention disclosed herein refers to or lists an “antibody,” it is intended to refer to any of the full-length antibodies or fragments thereof disclosed herein, unless expressly indicated otherwise.

A “humanized” antibody refers to an antibody in which some, most or all amino acids outside the CDR domains of a non-human antibody are replaced with corresponding amino acids derived from human immunoglobulins. In one embodiment of the humanized form of the antibody, some, most or all of the amino acids outside the CDR domains are replaced with amino acids from human immunoglobulin, while some, most or all amino acids within one or more CDR regions are unchanged. Small additions, deletions, insertions, substitutions or modifications of amino acids are permissible so long as they do not interfere with the antibody's ability to bind a particular antigen. A “humanized” antibody retains similar antigenic specificity to the original antibody.

A "chimeric antibody" refers to an antibody in which the variable regions are derived from one species and the constant regions are derived from another species, such as an antibody in which the variable regions are derived from a mouse antibody and the constant regions are derived from a human antibody.

"Complementarity determining region" or "CDR" refers to an amino acid sequence that together defines the binding affinity and specificity of the variable region of a native immunoglobulin binding site. There are three CDRs in each of the light and heavy chains of the antibody.

“Framework regions” or “FRs” refer to amino acid sequences intervening between typically conserved CDRs that act as a scaffold between the CDRs.

"Constant region" refers to the portion of an antibody molecule that is consistent for a particular class of antibody and is defined by the type of light and heavy chains. For example, the light chain constant region can be of kappa or lambda chain type and the heavy chain constant region can be of one of five chain isotypes: alpha, delta, epsilon, gamma or mu. Such constant regions are generally capable of conferring effector functions exhibited by the antibody. The different subclasses of heavy chains (such as the IgG subclass of heavy chains) are primarily responsible for different effector functions.

As used herein, a HER3 targeting agent includes, for example, an antibody as defined herein, e.g., a full-length antibody that binds with high immunoreactivity to any available epitope of HER3, such as human HER3, mini It can be a body or a nanobody.

As used herein, "immunoreactivity" refers to a measure of the ability of an immunoglobulin to recognize and bind to a specific antigen. “Specific binding” or “specifically binds” or “binds” means a greater affinity than for another antigen, e.g., within a relevant context, such as within the body of a mammalian subject, such as a human patient. It also refers to an antibody that binds an antigen or an epitope within an antigen. Antibodies that may be implemented or used in various aspects of the invention may have, for example, an equilibrium dissociation constant (K _D ) of about 1x10 ^-8 M or less, eg about 1x10 ^-9 M or less, about 1x10 ^-10 M or less, about Binds to an antigen or epitope within an antigen with a K _D of 1x10 ^-11 M or less, or about 1x10 ^-12 M or less, typically at least 100-fold lower than its K _D for binding to a non-specific antigen (eg BSA, casein) can do. Dissociation constants can be determined using standard procedures. However, an antibody that specifically binds an antigen or an epitope within an antigen is another related antigen, for example another species, such as human or monkey, for example Macaca fascicularis ( Macaca fascicularis ) (cynomolgus, cyno) , Pan troglodytes (chimpanzee, chimp) or Callithrix jacchus ) (common marmoset, marmoset) to the same antigen (homolog).

“Epitope” refers to a site on a target molecule (eg, at least a portion of an antigen) that can be recognized by and bound by a targeting agent, such as an antibody, antibody fragment, Fab fragment or aptamer. For example, in the case of a protein antigen, this can refer to the protein region (ie amino acids, and particularly side chains thereof) bound by the antibody. Overlapping epitopes contain at least 1 to 5 common amino acid residues. Methods for identifying epitopes of antibodies are known to those skilled in the art and include, for example, those described in Antibodies, A Laboratory Manual, Cold Spring Harbor Laboratory, Ed Harlow and David Lane (1988). .

Radiolabeled HER3 targeting agents as disclosed herein can be used to treat HER3-positive, ie, HER3-expressing cancers or precancerous conditions, such as solid tumors. “HER3-positive” or “HER3-expressing” means that at least some of the cancer cells in a patient, such as in a tumor, express or overexpress HER3.

As used herein, the terms “proliferative disorder” and “cancer” may be used interchangeably and include, but are not limited to, solid cancers (eg, tumors) and precancerous proliferative disorders. "Solid cancers" that may be HER3-positive and which may be treated by various aspects of the present invention include bone cancer, pancreatic cancer, skin cancer, head and neck cancer, cutaneous or intraocular malignant melanoma, uterine cancer, ovarian cancer, prostate cancer, rectal cancer, anal cancer. , gastric cancer, testicular cancer, uterine cancer, fallopian tube cancer, endometrial cancer, cervical carcinoma, vaginal carcinoma, vulvar carcinoma, esophageal cancer, small intestine cancer, endocrine cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, children tumor, bladder cancer, kidney or ureter cancer, renal pelvic carcinoma, neoplasia of the central nervous system (CNS), primary CNS lymphoma, tumor angiogenesis, spinal axis tumor, brainstem glioma, pituitary adenoma, Kaposi's sarcoma, epidermal cancer, squamous cell carcinoma, cancers caused by the environment including, but not limited to, cancers caused by asbestos. These cancers may be metastatic or non-metastatic.

According to certain aspects, the solid cancer that can be treated by various aspects of the present invention and which can be HER3-positive is breast cancer, such as tamoxifen-sensitive breast cancer, tamoxifen-resistant breast cancer, HER2-positive breast cancer, HER2-negative breast cancer or triple negative breast cancer. (TNBC), stomach cancer, bladder cancer, cervical cancer, endometrial cancer, skin cancer such as melanoma, stomach cancer, testicular cancer, esophageal cancer, bronchioloalveolar cancer, prostate cancer such as castration-resistant prostate cancer (CRPC), colorectal cancer, ovarian cancer, uterus Cervical epidermal cancer, liver cancer such as hepatocellular carcinoma (HCC) or cholangiocarcinoma, pancreatic cancer, lung cancer such as non-small cell lung carcinoma (NSCLC), kidney cancer, head and neck cancer such as head and neck squamous cell carcinoma, carcinoma, sarcoma, or any combination thereof. can be These cancers may be metastatic or non-metastatic.

According to certain aspects, the HER3 targeting agent may be labeled with a radioisotope/radionuclide. As used herein, a “radioisotope” or “radionuclide” may be an alpha emitting isotope, a beta emitting isotope, and/or a gamma emitting isotope. Exemplary radionuclides that can be used to label HER3 targeting agents or other targeting agents include: ¹³¹ I, ¹²⁵ I, ¹²³ I, ⁹⁰ Y, ¹⁷⁷ Lu, ¹⁸⁶ Re, ¹⁸⁸ Re, ⁸⁹ Sr, ¹⁵³ Sm, ³² P, ²²⁵ Ac, ²¹³ Bi, ²¹³ Po, ²¹¹ At, ²¹² Bi, ²¹³ Bi, ²²³ Ra, ²²⁷ Th, ¹⁴⁹ Tb, ¹³⁷ Cs, ²¹² Pb, ¹⁰³ Pd, ¹³⁴ Ce, ⁴³ Sc, ⁴⁴ Sc, ⁴⁷ Sc , ⁵⁵ Co, ⁶⁰ Cu, ⁶¹ Cu, ⁶² Cu, ⁶⁴ Cu, 67 Cu, ⁶⁶ Ga, ⁶⁷ Ga, ⁶⁸ Ga, ⁸² Rb, ⁸⁶ Y, ⁸⁷ Y, ⁸⁹ Zr, ⁹⁷ Ru ^{, 105 Rh} , ¹⁰⁹ Pd, ¹¹¹ In, ^117m Sn, ¹⁴⁹ Pm, ¹⁴⁹ Tb, ¹⁵³ Sm, ¹⁷⁷ Lu, ¹⁹⁹ Au, ²⁰¹ Tl, and ²⁰³ Pb. Methods for attaching radioisotopes to proteins, such as antibodies or antibody fragments (ie, "labeling" proteins with radioisotopes) are well known in the art. Specific compositions and methods for labeling are described, for example, in International Publication No. WO 2017/155937 and US Provisional Application No. 63/042,651 filed on December 9, 2019 and on November 30, 2020, "Site-specific radioactivity "Compositions and Methods for the Preparation of Conjugates" are disclosed in US Provisional Application Serial No. 63/119,093, filed in the name of the invention, each of which is incorporated herein by reference. HER3 targeting agents and other targeting agents containing one or more cysteine residues, such as peptides, proteins, antibodies and protein antibody mimetics, are "reagents for site-selective bioconjugation of proteins or antibodies", for example for radionuclide labeling. It may be chemically conjugated with any of the chelator-containing, such as DOTA-containing, stable linkers disclosed in U.S. Patent 11,000,604 entitled Iran Invention.

According to certain aspects, the HER3 targeting agent can be an antibody, peptide or small molecule that is radiolabeled with ²²⁵ Ac (" ²²⁵ Ac-labeled"), and the effective amount is, for example, 50.0 μCi/kg or less (i.e., administered to a subject where the amount of ²²⁵ Ac delivered delivers a radiation dose of less than 50.0 μCi per kilogram of the subject's body weight). According to certain aspects, when the HER3 targeting agent is ²²⁵ Ac-labeled, the effective amount is 50 μCi/kg, 40 μCi/kg, 30 μCi/kg, 20 μCi/kg, 10 μCi/kg, 5 μCi/kg, 4 μCi /kg, 3 μCi/kg, 2 μCi/kg, 1 μCi/kg, or less than 0.5 μCi/kg. According to certain aspects, when the HER3 targeting agent is ²²⁵ Ac-labeled, the effective amount is at least 0.05 μCi/kg, or 0.1 μCi/kg, 0.2 μCi/kg, 0.3 μCi/kg, 0.4 μCi/kg, 0.5 μCi/kg, 1 μCi/kg, 2 μCi/kg, 3 μCi/kg, 4 μCi/kg, 5 μCi/kg, 6 μCi/kg, 7 μCi/kg, 8 μCi/kg, 9 μCi/kg, 10 μCi/kg, 12 μCi/kg, 14 μCi/kg, 15 μCi/kg, 16 μCi/kg, 18 μCi/kg, 20 μCi/kg, 30 μCi/kg, or 40 μCi/kg. According to certain aspects, ^{the 225} Ac-labeled antibody is administered at a dose comprising any combination of the upper and lower limits as described herein, such as from at least 0.1 μCi/kg up to 5 μCi/kg, or from at least 5 μCi/kg to 20 μCi/kg. It can be administered in μCi/kg or less.

According to certain aspects, the HER3 targeting agent can be a ²²⁵ Ac-labeled antibody, peptide or small molecule, and the effective amount can be 2 mCi or less (i.e., wherein ^{the 225} Ac is administered to the subject in a non-weight based dose). . According to certain aspects, the effective dose of the ²²⁵ Ac-labeled HER3 targeting agent is less than 1 mCi, such as 0.9 mCi, 0.8 mCi, 0.7 mCi, 0.6 mCi, 0.5 mCi, 0.4 mCi, 0.3 mCi, 0.2 mCi, 0.1 mCi, 90 μCi, 80 μCi, 70 μCi, 60 μCi, 50 μCi, 40 μCi, 30 μCi, 20 μCi, 10 μCi, or 5 μCi. An effective amount of ²²⁵ Ac-labeled HER3 targeting agent is at least 2 μCi, such as at least 5 μCi, 10 μCi, 20 μCi, 30 μCi, 40 μCi, 50 μCi, 60 μCi, 70 μCi, 80 μCi, 90 μCi, 100 μCi, 200 μCi, 300 μCi, 400 μCi, 500 μCi, 600 μCi, 700 μCi, 800 μCi, 900 μCi, 1 mCi, 1.1 mCi, 1.2 mCi, 1.3 mCi, 1.4 mCi, or 1.5 mCi. According to certain aspects, ^{the 225} Ac-labeled HER3 targeting agent is at a dose comprising any combination of the upper and lower limits as described herein, e.g., at least 2 μCi up to 1 mCi, or at least 2 μCi up to 250 μCi; or between 75 μCi and up to 400 μCi.

According to certain aspects, ^{the 225} Ac-labeled HER3 targeting agent is administered to a subject at a dose of less than 12 Gy, or less than 8 Gy, or less than 6 Gy, or less than 4 Gy, or less than 2 Gy, such as from 2 Gy to 8 Gy; eg single doses delivered primarily to targeted solid tumors.

According to certain aspects, the HER3 targeting agent can be an antibody, peptide, or small molecule that has been radiolabeled with ¹⁷⁷ Lu (“ ¹⁷⁷ Lu-labeled”), and the effective amount can be, for example, less than 1 mCi/kg (i.e., where the amount of ¹⁷⁷ Lu-labeled antibody administered to the subject delivers a radiation dose of less than 1000 μCi per kilogram of the subject's body weight). According to certain aspects, when the antibody is ¹⁷⁷ Lu-labeled, the effective amount is 900 μCi/kg, 800 μCi/kg, 700 μCi/kg, 600 μCi/kg, 500 μCi/kg, 400 μCi/kg, 300 μCi/kg , 200 μCi/kg, 150 μCi/kg, 100 μCi/kg, 80 μCi/kg, 60 μCi/kg, 50 μCi/kg, 40 μCi/kg, 30 μCi/kg, 20 μCi/kg, 10 μCi/kg , 5 μCi/kg, or less than 1 μCi/kg. According to certain aspects, the effective amount of ^{the 177} Lu-labeled antibody is at least 1 μCi/kg, 2.5 μCi/kg, 5 μCi/kg, 10 μCi/kg, 20 μCi/kg, 30 μCi/kg, 40 μCi/kg, 50 μCi/kg, 60 μCi/kg, 70 μCi/kg, 80 μCi/kg, 90 μCi/kg, 100 μCi/kg, 150 μCi/kg, 200 μCi/kg, 250 μCi/kg, 300 μCi/kg, 350 μCi/kg, 400 μCi/kg or 450 μCi/kg. According to certain aspects, ^{the 177} Lu-labeled antibody is administered at a dose comprising any combination of the upper and lower limits as described herein, such as from at least 5 mCi/kg up to 50 μCi/kg, or from at least 50 mCi/kg to 500 μCi/kg. It can be administered in μCi/kg or less.

According to certain aspects, the HER3 targeting agent can be a ¹⁷⁷ Lu-labeled antibody, and the effective amount is less than 45 mCi, such as 40 mCi, 30 mCi, 20 mCi, 10 mCi, 5 mCi, 3.0 mCi, 2.0 mCi, 1.0 mCi, less than 800 μCi, 600 μCi, 400 μCi, 200 μCi, 100 μCi, or 50 μCi. An effective amount of ¹⁷⁷ Lu-labeled HER3 targeting agent is at least 10 μCi, such as at least 25 μCi, 50 μCi, 100 μCi, 200 μCi, 300 μCi, 400 μCi, 500 μCi, 600 μCi, 700 μCi, 800 μCi, 900 μCi, It may be 1 mCi, 2 mCi, 3 mCi, 4 mCi, 5 mCi, 10 mCi, 15 mCi, 20 mCi, 25 mCi, or 30 mCi. According to certain aspects, ^{the 177} Lu-labeled antibody is administered at a dose comprising any combination of the upper and lower limits as described herein, such as from at least 10 mCi up to 30 mCi, or from at least 100 μCi up to 3 mCi, or up to 3 mCi to 30 mCi or less.

According to certain aspects, the HER3 targeting agent can be an antibody, peptide, or small molecule that has been radiolabeled with ¹³¹ I (" ¹³¹ I-labeled"), and the effective amount can be, for example, less than 1200 mCi (i.e., to a subject if the amount of ¹³¹ I administered delivers a total body radiation dose of less than 1200 mCi in a dose that is not based on body weight). According to certain aspects, the effective amount of the ¹³¹ I-labeled targeting agent is less than 1100 mCi, less than 1000 mCi, less than 900 mCi, less than 800 mCi, less than 700 mCi, less than 600 mCi, less than 500 mCi, less than 400 mCi, less than 300 mCi. , less than 200 mCi, less than 150 mCi or less than 100 mCi. According to certain aspects, the effective amount of the ¹³¹ I-labeled targeting agent is less than 200 mCi, such as 190 mCi, 180 mCi, 170 mCi, 160 mCi, 150 mCi, 140 mCi, 130 mCi, 120 mCi, 110 mCi, 100 mCi, It may be less than 90 mCi, 80 mCi, 70 mCi, 60 mCi, or 50 mCi. According to certain aspects, the effective amount of the ¹³¹ I-labeled targeting agent is at least 1 mCi, such as at least 2 mCi, 3 mCi, 4 mCi, 5 mCi, 6 mCi, 7 mCi, 8 mCi, 9 mCi, 10 mCi, 20 mCi , 30 mCi, 40 mCi, 50 mCi, 60 mCi, 70 mCi, 80 mCi, 90 mCi, 100 mCi, 110 mCi, 120 mCi, 130 mCi, 140 mCi, 150 mCi, 160 mCi, 170 mCi, 180 mCi , 190 It may be mCi, 200 mCi, 250 mCi, 300 mCi, 350 mCi, 400 mCi, 450 mCi, or 500 mCi. According to certain aspects, ^{the 131} I-labeled targeting agent can be administered at a dose comprising any combination of the upper and lower limits as described herein, such as between at least 1 mCi and up to 100 mCi, or between at least 10 mCi and up to 200 mCi. can

Although selected radionuclides are discussed in detail herein, any radionuclide, such as any disclosed herein, may be used with a radiolabeled targeting agent, such as a radiolabeled HER3 targeting agent as disclosed herein.

As used herein, a composition comprising a HER3 targeting agent includes a "patient specific composition" comprising both a radionuclide labeled and unlabeled portion. According to certain aspects of the present invention, when the HER3 targeting agent is labeled with a radioisotope, the majority of the targeting agent (antibody, antibody fragment, etc.) administered to the patient may consist of unlabeled targeting agent, and a minority may consist of radiolabeled targeting agent. It is a targeting agent. The ratio of labeled to unlabeled targeting agent can be adjusted using known methods. According to certain aspects of the present invention, the patient specific composition comprises a ratio of about 0.01:10 to 1:1, such as 0.1:10 to 1:1 labeled:unlabeled labeled HER3-targeting agent:unlabeled HER3-targeting agent. HER3 targeting agents.

According to certain aspects of the invention, the HER3 targeting agent is in a total protein or peptide amount of at most 100 mg, eg at most 60 mg, such as 5 mg to 45 mg, or 0.001 mg/kg (patient body weight) to 3.0 mg/kg (patient body weight), such as 0.005 mg/kg (patient body weight) to 2.0 mg/kg (patient body weight), or 0.01 mg/kg (patient body weight) to 1 mg/kg (patient body weight), or 0.1 mg/kg (patient body weight) body weight) to 0.6 mg/kg (patient body weight), or 0.3 mg/kg (patient body weight), or 0.4 mg/kg (patient body weight), or 0.5 mg/kg (patient body weight), or 0.6 mg/kg (patient body weight) ) can be provided with a total protein amount of

The present combination of radiolabeled and unlabeled fractions of an antibody or other targeting agent allows compositions to be tailored to a specific patient, wherein each of the protein dose and the radiation dose of the antibody or other targeting agent is at least one It is personalized to that patient based on patient-specific parameters. As such, each vial of the composition can be prepared for a specific patient, wherein the entire contents of the vial are delivered to that patient in a single dose. When a treatment regimen calls for multiple doses, each dose may be formulated as a patient specific dose in a vial and administered to the patient as a “single dose” (i.e., the entire contents of the vial are administered at one time). ). Subsequent doses may be formulated in a similar manner, such that each dose in the regimen provides a patient specific dose in a single dose container. One of the advantages of such a composition is that there will be no residual radiation that needs to be discarded or handled by medical personnel, eg, no dilution or other manipulations to obtain a dose for a patient. When provided in a single dose container, the container can simply be placed in-line with a set of infusion tubing for infusion into a patient. Furthermore, volume can be standardized to greatly reduce the possibility of medical error (ie, delivery of an incorrect dose, since the entire volume of the composition must be administered in one infusion).

Thus, according to certain aspects, the HER3 targeting agent can be provided as a single dose composition that can be tailored to a specific patient, wherein the amount of radiolabeled and unlabeled HER3 targeting agent in the composition is dependent on the patient's weight, age, sex, One or more of the disease state and/or health condition, such as those detailed in International Publication No. WO 2016/187514 and US Patent No. 10,736,975. According to certain aspects, the HER3 targeting agent may be provided as a multi-dose therapeutic, wherein each dose in the treatment regimen is provided as a patient specific composition. The patient-specific composition comprises radiolabeled and unlabeled HER3 targeting agents, wherein the amount of each varies according to one or more of the patient's weight, age, sex, disease state, and/or health condition.

As used herein, the terms “subject” and “patient” are interchangeable and refer to mammals such as humans, non-human primates, dogs, cats, horses, sheep, goats, cows, rabbits, pigs, rats and mice. Including, but not limited to. Where the subject is a human, such subject may be of any age. For example, the subject may be 60 years of age or older, 65 years of age or older, 70 years of age or older, 75 years of age or older, 80 years of age or older, 85 years of age or older, or 90 years of age or older. Alternatively, the subject may be under 50 years old, under 45 years old, under 40 years old, under 35 years old, under 30 years old, under 25 years old, or under 20 years old. In the case of a human subject with cancer, such subject may be newly diagnosed, relapsed and/or refractory, or in remission.

As used herein, "treating" a subject suffering from cancer refers to (i) slowing, stopping or reversing the progression of cancer, (ii) slowing, stopping or reversing the progression of symptoms of cancer. or reversing, (iii) reducing the likelihood of cancer recurrence, and/or (iv) reducing the likelihood of cancer symptoms recurring. According to certain preferred aspects, treating a subject suffering from cancer (i) reverses the progression of the cancer, ideally to the point of eliminating the cancer, and/or (ii) ideally to the point of eliminating the symptoms. reversing the progression of cancer symptoms, and/or (iii) reducing or eliminating the likelihood of recurrence (i.e., integration ideally resulting in destruction of any remaining cancer cells).

A “chemotherapeutic agent” in the context of the present invention means a chemical compound that inhibits or kills growing cells and can be used or is approved for use in the treatment of cancer. Exemplary chemotherapeutic agents include cytostatic agents that prevent, disrupt, hinder or retard cell division at the level of nuclear division or cell plasma division. Such agents, such as taxanes, especially docetaxel or paclitaxel, and epothilones, especially epothilones A, B, C, D, E and F, can stabilize microtubules, or, such as vinca alkaloids, especially vinblastine, vincristine , Vindesine, vinflunine and vinorelbine can destabilize microtubules. Exemplary chemotherapeutic agents such as temozolomide, cisplatin and/or fluorouracil also include radiosensitizers that can synergize with radiolabeled HER3.

“Therapeutically effective amount” or “effective amount” refers to an amount effective at dosages and for periods of time necessary to achieve the desired therapeutic result. A therapeutically effective amount may vary depending on factors such as the disease state, age, sex, and weight of the individual, and the ability of the therapeutic agent or combination of therapeutic agents to elicit a desired response in the individual. Exemplary indicators of an effective therapeutic agent or therapeutic combination include, for example, improved patient well-being, reduced tumor burden, stopped or slowed growth of tumors, and/or absence of metastasis of cancer cells to other sites in the body. According to certain aspects, a "therapeutically effective amount" or "effective amount", when used alone or in combination or conjunction with other agents and/or treatment modalities, is capable of depleting or causing a decrease in the total number of cells expressing HER3. and/or capable of inhibiting the growth of cells expressing HER3.

As used herein, “depleting” with respect to cells that express HER3 means at least one type of cell that expresses or overexpresses HER3 (e.g., HER3 circulating in a solid tumor or in the blood of a subject). -positive cells) to lower the population. According to certain aspects of the invention, the reduction is determined by comparing the number of HER3-positive cells in a blood or tissue biopsy of the subject, such as from a solid tumor, before and after initiation of treatment with the HER3 targeting agent. Thus, for example, a subject's HER3-positive cells are reduced in that population by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 99%. , can be considered depleted.

"Inhibit growth" means that the growth of malignant cells or tissues (eg, tumors) in vitro or in vivo when contacted with a therapeutic agent or combination of therapeutic agents or drugs, in the absence of the therapeutic agent or combination of therapeutic drugs, inhibits the growth of the same cells or a measurable decrease or retardation when compared to a decrease or retardation in the growth of a tissue. Inhibition of growth of malignant cells or tissues in vitro or in vivo by at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97% , 98%, 99%, or 100%.

The term “immune checkpoint therapy” refers to a molecule capable of modulating, in a positive or negative manner, the function of an immune checkpoint protein in promoting an immune response against cancer cells. The term "immune checkpoint" refers to proteins involved directly or indirectly in immune pathways that act to prevent uncontrolled immune responses under normal physiological conditions to maintain self-tolerance and/or tissue protection.

In the context of the present invention, an immune checkpoint therapy can at least partially upregulate the function of an antibody and/or a stimulatory immune checkpoint (agonist) that can at least partially downregulate the function of an inhibitory immune checkpoint (antagonist). It encompasses therapies such as antibodies in For example, immune checkpoint therapy can refer to antibodies against immune checkpoint inhibitors (ICIs) that can be upregulated in certain cancers and thus inhibit the function of ICIs.

The term “DDRi” refers to inhibitors of DNA damage response pathway proteins, eg PARPi. The term “PARPi” refers to inhibitors of poly(ADP-ribose) polymerase. In the context of the present invention, the term PARPi encompasses molecules capable of binding to and inhibiting the function of poly(ADP-ribose) polymerase, such as antibodies, peptides or small molecules.

The term "CD47 blocker" refers to an agent that prevents CD47 binding to SIRPα, such as a blocker that binds either CD47 or SIRPα, or a blocker that modulates the expression of CD47 or SIRPα, or a blocker that otherwise inhibits the CD47/SIRPα axis. refers to Without limitation, CD47 blockers modulate the expression of antibodies that bind at least CD47, such as magrolimab, remzoparlimab, and AO-176, SIRPα fusion proteins such as TTI-621 and TTI-622, CD47 and/or SIRPα. agents that block the translation of CD47, such as phosphorodiamidate morpholino oligomers (PMOs), and small molecule agents, such as RRx-001.

As used herein, administration of one or more additional therapies, such as immune checkpoint therapy and/or one or more of DDRi and/or CD47 blockers and/or radiosensitizers to a subject "in conjunction with" a HER3 targeting agent It means administering an additional therapy before, during and/or after administration of the HER3 targeting agent. Such administration includes, but is not limited to, the following scenarios: (i) the additional therapy is administered first and the HER3 targeting agent is administered second; (ii) the additional therapy is administered concurrently with the HER3 targeting agent (e.g., DDRi is administered orally once daily for n days, and the HER3 targeting agent is administered on days 2 to n-1 of the DDRi regimen). administered intravenously as a single dose on day 1 of (iii) the additional therapy is administered concurrently with the HER3 targeting agent (e.g., the DDRi is administered orally for a duration of greater than 1 month, e.g., 35 days, 42 days, 49 days, or progression of the cancer under treatment administered orally once daily for a longer period of time where the DDRi does not cause unacceptable toxicity, and the HER3-targeting agent is administered intravenously as a single daily dose within the first month of DDRi therapy); and (iv) the HER3 targeting agent is administered first (e.g., administered intravenously in a single dose or in multiple doses over a period of several weeks) and the additional therapy is administered second (e.g., For example, DDRi is administered orally once daily for 21 days, 28 days, 35 days, 42 days, 49 days, or a longer period during which the cancer under treatment does not progress and the DDRi does not cause unacceptable toxicity). Additional permutations obvious to those skilled in the art are possible and are within the scope of the invention claimed by the present application.

"Article of manufacture" refers to a package containing materials useful for the treatment, prevention and/or diagnosis of the disorders described herein. Articles of manufacture include containers; and a label or package insert on or associated with the container. Suitable containers include, for example, bottles, vials, syringes, IV solution bags, and the like. The container may be formed from a variety of materials, such as glass or plastic. The container holds a composition by itself or in combination with another composition effective for treating, preventing and/or diagnosing a condition, and can have a sterile access port (eg, the container can be an intravenous solution bag or a hypodermic needle It may be a vial with a stopper pierceable by). At least one active agent in the composition may be a radiolabeled HER3 targeting agent according to aspects of the invention disclosed herein.

"Labels" or "package inserts" are customarily included in commercial packages of therapeutic products that contain information about indications, directions for use, dosage, administration, combination therapy, contraindications and/or warnings regarding the use of such therapeutic products. It is used to refer to a set of guidelines. As used herein, a label may indicate that the composition is used to treat HER3-positive cancer, and may optionally indicate a route and/or method of administration. Moreover, the article of manufacture may comprise (a) a first container containing therein a composition comprising a HER3 targeting agent; and (b) a second container containing therein a composition comprising an additional cytotoxic or other therapeutic agent according to aspects of the invention disclosed herein. Alternatively or additionally, the article of manufacture may further comprise a second (or third) container comprising a pharmaceutically acceptable buffer, such as bacteriostatic water for injection (BWFI), phosphate buffered saline, Ringer's solution and dextrose solution. there is. It may further include other materials desirable from a commercial and user standpoint, including other buffers, diluents, filters, needles, and syringes.

Throughout this application, various patents, patent applications and other publications are cited, each of which is incorporated herein by reference in its entirety.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing described herein, suitable methods and materials are described below.

Experiment result

An anti-HER3 IgG monoclonal antibody consisting of heavy chain SEQ ID NO: 77 and light chain SEQ ID NO: 78 was prepared, conjugated to the chelator DOTA using p-SCN-Bn-DOTA, and actinium Radiolabeling via chelation to -225 was further investigated as described below in conjunction with Figures 3-11.

Figure 3 shows the ELISA assay binding characteristics of an Ac225 labeled DOTA-conjugated anti-HER3 monoclonal antibody ("HER3-ARC") compared to an unmodified anti-HER3 antibody and an unspecific antibody (IgG), indicating that such It demonstrates that the modification does not substantially affect immune reactivity to HER3.

The binding properties of 225Ac-HER3-ARC were evaluated by ELISA. A 96-well plate was coated overnight with human recombinant HER3, followed by serial dilutions (0 - 100 μg/ml) of anti-HER3, 225Ac-HER3-ARC and IgG (immunoglobulin 1, non-specific IgG1 control) for 1 hour at room temperature. incubated during. A secondary antibody (goat anti-human IgG F(ab')20-HRP) was added and incubated on ice for 30 minutes, followed by development with HCl 1M for 10 minutes. Sample absorbance was measured at 450 nm. ²²⁵ Ac-HER3-ARC showed similar binding properties to native antibodies by ELISA (HER3-ARC: EC ₅₀ = 0.0017 μg/ml, HER3 EC ₅₀ = 0.0022 μg/ml).

Figure 4 shows 225Ac-HER3-ARC, unmodified anti-HER3 mAb, non-specific antibody control (IgG) and BxPC-3 cells (human pancreatic adenocarcinoma) against HER3-positive NCI-H1975 cells (human lung adenocarcinoma, NSCLC) and BxPC-3 cells (human pancreatic adenocarcinoma). A graph showing the results of a flow cytometry assay examining the binding of the secondary antibody only control.

The binding properties of 225Ac-HER3-ARC were evaluated by flow cytometry in HER3+ cells (NCI-H1975 and BxPC3). A solution (100 μg/ml) of anti-HER3, 225Ac-HER3-ARC and IgG (immunoglobulin 1, non-specific IgG1) was added to HER+ cells and incubated for 1 hour at room temperature. PE-labeled secondary antibody was added and incubated on ice for 30 minutes. Sample fluorescence was measured using a flow cytometer. The binding properties of 225Ac-HER3-ARC to HER3+ positive cell lines were similar to those of the unmodified anti-HER3 mAb.

5 is a graph showing the in vitro cytotoxic effect of 225Ac-HER3-ARC on the HER3-positive cell line NCI-H1975 as a function of radiation dose.

The cytotoxic effect of 225Ac-HER3-ARC on the HER3+ cell line NCI-H1975 was evaluated in a colorimetric assay using the CellTiter 96® aqueous non-radioactive cell proliferation assay (MTS). NCI-H1975 cells were incubated with 225Ac-HER3-ARC for 24 hours at 37°C. Unbound 225Ac-HER3-ARC was then removed and the cells were cultured at 37°C for 72 hours. The absorbance at 490 nm was measured and the % of cell viability was calculated. 225Ac-HER3-ARC showed potent in vitro cytotoxicity against the HER3+ cell line NCI-H1975.

Figure 6A is a graph showing that 225Ac-HER3-ARC upregulates cell surface calreticulin (CRT) in NCI-H1975 cells and Figure 6B shows that 225Ac-HER3-ARC upregulates CD47 on NCI-H1975 cells. This is the graph that shows

The effect of 225Ac-HER3-ARC on the cell surface expression of calreticulin (CRT) and CD47 by the HER3+ cell line NCI-H1975 was investigated using flow cytometry. Cells were treated with 225Ac-HER3-ARC (100 nCi/ml) or PBS (control) for 72 hours. After treatment, cells were stained for CRT and CD47. The results demonstrate that each of CRT (FIG. 6A) and CD47 (FIG. 6B) is upregulated by 225Ac-HER3-ARC in NCI-H1975 cells.

7A is a graph showing the results of a phagocytosis assay demonstrating that the combination of 225Ac-HER3-ARC with an anti-CD47 blocking antibody enhanced phagocytosis of BxPC-3 cells compared to either treatment alone. 7B is a graph showing the results of a phagocytosis assay demonstrating that the combination of 225Ac-HER3-ARC with an anti-CD47 blocking antibody enhanced phagocytosis of NCI-H1975 cells compared to either treatment alone. The same keys apply to Figures 7A and 7B.

The effect of the combination of 225Ac-HER3-ARC and anti-CD47 on phagocytosis in vitro was evaluated by flow cytometry. BxPC-3 (FIG. 7A) and NCI-H1975 (FIG. 7B) cells were seeded in 6-well plates 24 hours before incubation with 225Ac-HER3-ARC for 24 hours at 37°C. After treatment with 225Ac-HER3-ARC, cells were cultured at 37°C for 72 hours.

BxPC-3 and NCI-H1975 cells were stained with Vybrant DiD cell-labeling solution and stained with anti-human CD47 (Bio X Cell, Cat#BE0019) and mouse IgG1 isotype control (Bio X Cell). , Cat#BE0083) at 37°C for 1 hour. Human macrophages were stained with Vibrandt DiO cell-labeling solution. Labeled human macrophages and target cells were co-cultured at 37°C for 2 hours. Phagocytosis was assessed by evaluating dual labeled cells (DiD+/DiO+).

Figure 8: 225Ac-HER3- in combination with anti-CD47 blocking antibody at different radiation doses (100 nCi, 200 nCi, 400 nCi, 600 nCi) alone and at 200 nCi in a human tumor (NCI-H1975 cell) mouse xenograft model. A graph showing the effect of ARC, unlabeled anti-HER3 mAb, anti-CD47 blocking antibody alone, and vehicle-only control on tumor growth. In particular, tumor growth was almost completely inhibited by 225Ac-HER-ARC at each radiation dose of 200 nCi, 400 nCi, 600 nCi and by the combination of 225Ac-HER-ARC (200 nCi) with anti-CD47 mAb.

9 is a graph showing body weight over time for the subjects of the experiment described in FIG. 8 .

10 is a graph showing survival probability over time for subjects in the experimental group of the experiment described in FIG. 8 .

A tumor xenograft study was also conducted examining the effect of HER2-ARC treatment alone and in combination with a CD47 blocker on HER2-positive tumor growth. The anti-HER2 mAb Trastuzumab was chemically conjugated to DOTA using p-SCN-Bn-DOTA and labeled with actinium-225 or lutetium-177 via chelation for use in these experiments.

Figure 11 shows vehicle alone (control), magnolimab alone (10 mg/kg), 225Ac-trastuzumab alone (0.025 μCi/animal) in an NGS mouse xenograft model using the HER2-positive SK-OV3 human ovarian cancer cell line. , and Magrolimab (10 mg/kg) and 225Ac-Trastuzumab (0.025 μCi/animal) in combination with comparative effects on tumor growth. Each cohort consisted of 8 animals.

Figure 12 shows vehicle alone (control), magnolimab alone (10 mg/kg), 177Lu-trastuzumab alone (25 μCi/animal) in an NGS mouse xenograft model using HER2-positive SK-OV3 human ovarian cancer cell line. , and Magrolimab (10 mg/kg) and 177Lu-Trastuzumab (25 μCi/animal) are graphs showing comparative effects on tumor growth. Each cohort consisted of 8 animals.

aspect of the invention

It is well documented in both preclinical and clinical studies that HER3 levels can be downregulated following administration of HER3-targeting antibodies (Mishra, 2018). In preclinical models using rumletuzumab, there was a dose-dependent (1-10 mg/kg) downregulation of HER3 as measured by both immunohistochemistry and western blotting (Maneses-Lorenta, 2015; Mirshberger, 2013). The lowest dose of rumletuzumab (0.3 mg/kg) did not result in HER3 target downregulation (Maneses-Lorenta, 2015), and this low dose of rumletuzumab (0.1 mg/kg and 0.3 mg/kg) did not result in downregulation of the HER3 target. kg) was not effective in controlling HER3-expressing tumors (Mirshberger, 2013). In a clinical study with rumletuzumab, downregulation of surface HER3 was observed in serial tumor biopsies in 92% of patients across all dose levels tested (100-2000 mg; Meulendijks, 2016). Additionally, reductions in total HER3 levels were observed in 3 out of 5 paired tumor biopsy samples from patients treated with 40 mg/kg of the HER3-targeting antibody LJM716 (Reynolds, 2017).

Internalization and degradation of HER3 can be beneficial in reducing phosphorylation and subsequent signaling activity of HER3, but reduced surface levels of HER3 can interfere with antibody targeting in tumors. Thus, if repeated administration of the HER3-targeting antibody is desired or necessary for efficacy, administration of the HER3-targeting antibody may result in downregulation of the target and preclude re-administration. The inventors have discovered that using an antibody radioconjugate (ARC) avoids the problems associated with dose-dependent downregulation of HER3 because lower antibody doses useful in therapeutic methods may not result in HER3 downregulation. Accordingly, the present inventors have found that HER3 targeting agents including radioisotopes are effective as diagnostic and therapeutic agents for improved tumor targeting and killing of HER3-expressing cancer cells, such as certain solid tumors. In particular, treatment methods that can include multiple doses of HER3-targeting agents can provide improved tumor targeting and killing without causing detrimental levels of HER3 downregulation.

Accordingly, according to certain aspects of the invention disclosed herein, therapeutic methods are provided for treating HER3-positive cancers using radiolabeled HER3 targeting agents. Such methods may also be used to determine whether and/or to what extent a patient has a HER3-positive cancer, for example, by identifying and/or quantifying HER3 positive cells circulating within a solid tumor or in a blood sample from the patient. diagnostic steps to determine whether and/or the localization of such cancer.

According to certain aspects, the method of treatment comprises administering a radiolabeled HER3 targeting agent, such as a radiolabeled antibody, peptide or small molecule targeting HER3, alone or in combination with one or more additional therapeutic agents or treatment modalities. According to certain aspects, the additional agent or modality may be administration of any one or more of immune checkpoint therapy, DDRi, CD47 blockers, chemotherapeutic agents, small molecule oncology drugs, external beam radiation, and brachytherapy.

According to certain aspects, the radiolabeled HER3 targeting agent may be administered to a patient in one or more doses in a patient specific composition.

According to certain aspects, the patient may be monitored at intervals during therapy for the presence of HER3-positive cells to assess a decrease in HER3-positive cells. Detecting a reduced number of HER3-positive cells after treatment with a HER3-targeting agent, as compared to the number of HER3-positive cells prior to treatment, improves the effectiveness of the HER3-targeting agent in treating HER3-positive cancer in a mammalian subject. can indicate

According to certain aspects, a method of treating cancer comprises identifying a patient having a HER3-positive cancer by identifying HER3-positive cells, and administering to the patient an effective amount of a HER3 targeting agent, alone or in combination with an additional treatment method. include According to certain aspects, the additional treatment method may be administration of any one or more of immune checkpoint therapy, DDRi, CD47 blockers, chemotherapeutic agents, small molecule oncology drugs, external beam radiation, and brachytherapy.

According to certain aspects, the chemotherapeutic agent is a radiosensitizer.

According to certain aspects, a radiolabeled HER3 targeting agent can be administered to a patient who has undergone, eg, has recently undergone, surgery for treatment, eg, cancer treatment, such as removal of all or part of a solid tumor. Thus, for example, a radiolabeled HER3 targeting agent can be administered before or after surgery.

HER3 targeting agent

It is an object of the invention disclosed herein to provide radiolabeled HER3-targeting agents, such as human HER3-targeting agents, for diagnostic and/or therapeutic uses, such as in the diagnosis and/or treatment of HER3-positive cancers. Radiolabeled HER3-targeting agents can elicit a therapeutic response through the delivery of DNA-damaging ionizing radiation, eg, alpha particles to cells that induce double-stranded DNA breakage and cell death.

Exemplary anti-HER3 antibodies (also referred to herein as “HER3 antibodies”) that are radiolabeled and can be implemented and/or used in various aspects of the invention disclosed herein, such as anti-human HER3 antibodies, include the following antibodies, and antibodies , such as, but not limited to, immunoglobulins, such as, but not limited to, IgG, including but not limited to: (i) those comprising a heavy chain variable region of a HER3 antibody or heavy chain, (ii) HER3 one, two or three heavy chain CDRs of an antibody or heavy chain (e.g., by the Kabat definition) or those recited, (iii) one comprising the light chain variable region of a HER3 antibody or light chain, and /or (iv) one, two or three light chain CDRs of a HER3 antibody or light chain (eg, according to the Kabat definition) or those recited. Furthermore, where a HER3 antibody heavy chain or HER3 antibody light chain comprising an N-terminal leader sequence is disclosed, it is also intended to be disclosed for embodiments and use in the various aspects of the present invention that the corresponding HER3 antibody lacking a leader sequence is also disclosed. It should also be understood that heavy chains and corresponding light chains.

Exemplary HER3 antibodies that are radiolabeled and can be implemented and/or used in the invention disclosed herein include a heavy chain having an amino acid sequence as set forth in, for example, SEQ ID NO: 9 or 11 and/or SEQ ID NO: 10 or 12, a murine monoclonal antibody to HER3 comprising a light chain having the amino acid sequence as set forth in , or an antibody such as a humanized antibody derived from one or more of the foregoing sequences. Exemplary HER3 antibodies that are radiolabeled and can be implemented and/or used in the invention disclosed herein include a heavy chain having an N-terminal region having the sequence set forth in SEQ ID NO:13 and/or as set forth in SEQ ID NO:14. and a light chain having an N-terminal region having the sequence. HER3 antibodies that can be similarly implemented or used in various aspects of the present invention include, for example, a heavy chain variable region having an amino acid sequence as set forth in SEQ ID NO:7, and/or an amino acid sequence as set forth in SEQ ID NO:8. a light chain variable region having the sequence; and/or a heavy chain comprising one or more of CDR1, CDR2 and CDR3 having the amino acid sequences set forth in SEQ ID NOs: 1-3, respectively, and/or CDR1, CD2 and CDR1 having the amino acid sequences set forth in SEQ ID NOs: 4-6, respectively. and a light chain comprising one or more of the CDR3s. See Figures 1 and 2 for further description of these sequences. A HER3 antibody implemented and/or used in any of the aspects of the present invention may comprise, for example, any combination of light chain sequences and/or heavy chain sequences described above.

An exemplary HER3 antibody comprises an immunoglobulin heavy chain variable region comprising CDR-H1 comprising SEQ ID NO: 15, CDR-H2 comprising SEQ ID NO: 16, and CDR-H3 comprising SEQ ID NO: 17 , and/or an immunoglobulin light chain variable region comprising CDR-L1 comprising SEQ ID NO: 18, CDR-L2 comprising SEQ ID NO: 19, and CDR-L3 comprising SEQ ID NO: 20; include An exemplary HER3 antibody comprises an immunoglobulin heavy chain variable region comprising SEQ ID NO:21 and/or an immunoglobulin light chain variable region comprising SEQ ID NO:22. An exemplary HER3 antibody comprises the immunoglobulin heavy chain amino acid sequence of SEQ ID NO:23 and/or the immunoglobulin light chain amino acid sequence of SEQ ID NO:24.

An exemplary HER3 antibody comprises an immunoglobulin heavy chain variable region comprising CDR-H1 comprising SEQ ID NO: 25, CDR-H2 comprising SEQ ID NO: 26, and CDR-H3 comprising SEQ ID NO: 27 ; and/or an immunoglobulin light chain variable region comprising CDR-L1 comprising SEQ ID NO: 28, CDR-L2 comprising SEQ ID NO: 29, and CDR-L3 comprising SEQ ID NO: 30. do. An exemplary HER3 antibody comprises an immunoglobulin heavy chain variable region comprising SEQ ID NO:31 and/or an immunoglobulin light chain variable region comprising SEQ ID NO:32. An exemplary HER3 antibody comprises the immunoglobulin heavy chain amino acid sequence of SEQ ID NO:33 and/or the immunoglobulin light chain amino acid sequence of SEQ ID NO:34.

An exemplary HER3 antibody comprises an immunoglobulin heavy chain variable region comprising CDR-H1 comprising SEQ ID NO: 35, CDR-H2 comprising SEQ ID NO: 36, and CDR-H3 comprising SEQ ID NO: 37 ; and/or an immunoglobulin light chain variable region comprising CDR-L1 comprising SEQ ID NO: 38, CDR-L2 comprising SEQ ID NO: 39, and CDR-L3 comprising SEQ ID NO: 40. do. An exemplary HER3 antibody comprises an immunoglobulin heavy chain variable region comprising SEQ ID NO:41 and/or an immunoglobulin light chain variable region comprising SEQ ID NO : 42. An exemplary HER3 antibody comprises the immunoglobulin heavy chain amino acid sequence of SEQ ID NO:43 and the immunoglobulin light chain amino acid sequence of SEQ ID NO:44.

An exemplary HER3 antibody comprises an immunoglobulin heavy chain variable region comprising CDR-H1 comprising SEQ ID NO:45, CDR-H2 comprising SEQ ID NO:46, and CDR-H3 comprising SEQ ID NO:47 ; and/or an immunoglobulin light chain variable region comprising CDR-L1 comprising SEQ ID NO:48, CDR-L2 comprising SEQ ID NO:29, and CDR-L3 comprising SEQ ID NO:49. do. An exemplary HER3 antibody comprises an immunoglobulin heavy chain variable region comprising SEQ ID NO:50 and/or an immunoglobulin light chain variable region comprising SEQ ID NO:51. An exemplary HER3 antibody comprises the immunoglobulin heavy chain amino acid sequence of SEQ ID NO:52 and/or the immunoglobulin light chain amino acid sequence of SEQ ID NO:53.

An exemplary HER3 antibody comprises an immunoglobulin heavy chain variable region comprising CDR-H1 comprising SEQ ID NO:54, CDR-H2 comprising SEQ ID NO:55, and CDR-H3 comprising SEQ ID NO:56 ; and/or an immunoglobulin light chain variable region comprising CDR-L1 comprising SEQ ID NO: 28, CDR-L2 comprising SEQ ID NO: 29, and CDR-L3 comprising SEQ ID NO: 30. do. An exemplary HER3 antibody comprises an immunoglobulin heavy chain variable region comprising SEQ ID NO:57 and/or an immunoglobulin light chain variable region comprising SEQ ID NO:58. An exemplary HER3 antibody comprises the immunoglobulin heavy chain amino acid sequence of SEQ ID NO:59 and/or the immunoglobulin light chain amino acid sequence of SEQ ID NO:60.

An exemplary HER3 antibody comprises an immunoglobulin heavy chain variable region comprising CDR-H1 comprising SEQ ID NO: 61, CDR-H2 comprising SEQ ID NO: 62, and CDR-H3 comprising SEQ ID NO: 63 ; and/or an immunoglobulin light chain variable region comprising CDR-L1 comprising SEQ ID NO:64, CDR-L2 comprising SEQ ID NO:65, and CDR-L3 comprising SEQ ID NO:66 do. An exemplary HER3 antibody comprises an immunoglobulin heavy chain variable region comprising SEQ ID NO:67 and/or an immunoglobulin light chain variable region comprising SEQ ID NO:68. An exemplary HER3 antibody comprises the immunoglobulin heavy chain amino acid sequence of SEQ ID NO:69 and the immunoglobulin light chain amino acid sequence of SEQ ID NO:70.

An exemplary HER3 antibody comprises an immunoglobulin heavy chain variable region comprising CDR-H1 comprising SEQ ID NO: 71, CDR-H2 comprising SEQ ID NO: 72, and CDR-H3 comprising SEQ ID NO: 66 ; and/or an immunoglobulin light chain variable region comprising CDR-L1 comprising SEQ ID NO: 28, CDR-L2 comprising SEQ ID NO: 29, and CDR-L3 comprising SEQ ID NO: 30. do. An exemplary HER3 antibody comprises an immunoglobulin heavy chain variable region comprising SEQ ID NO:73 and/or an immunoglobulin light chain variable region comprising SEQ ID NO:74. An exemplary HER3 antibody comprises the immunoglobulin heavy chain amino acid sequence of SEQ ID NO:75 and/or the immunoglobulin light chain amino acid sequence of SEQ ID NO:76.

An exemplary HER3 antibody comprises the immunoglobulin heavy chain amino acid sequence of SEQ ID NO:77 and/or the immunoglobulin light chain amino acid sequence of SEQ ID NO:78.

Exemplary HER3 antibodies comprise an immunoglobulin light chain variable region comprising SEQ ID NOs: 86, 87, 88, 89, 90 or 91 and/or SEQ ID NOs: 79, 80, 81, 82, 83, 84 or 85. A heavy chain variable region comprising

An exemplary HER3 antibody comprises an immunoglobulin heavy chain sequence comprising SEQ ID NOs: 92, 94, 95, 98 or 99 and/or an immunoglobulin light chain sequence comprising SEQ ID NOs: 93, 96, 97, 100 or 101 includes

Exemplary HER3 antibodies also include baresetamab (ISU104) from Isu Abxis Company and any of the HER3 antibodies disclosed in US Pat. No. 10,413,607.

Exemplary HER3 antibodies are also described in HMBD-001 (10D1F) from Hummingbird Biosciences Pte. and International Publication Nos. WO 2019185164 and WO2019185878, U.S. Patent Nos. 10,662,241; and any of the HER3 antibodies disclosed in US Publication Nos. 20190300624, 20210024651, and 20200308275.

Exemplary HER3 antibodies also include the HER2/HER3 bispecific antibody MCLA-128 from Merus N.V. (ie, Genocetuzumab); and any of the monospecific or multispecific HER3 antibodies disclosed in US Publication Nos. 20210206875, 20210155698, 20200102393, 20170058035, and 20170037145.

Exemplary HER3 antibodies also include the HER3 antibody patritumab (U3-1287), an antibody comprising the heavy chain sequence SEQ ID NO: 106 and/or light chain sequence SEQ ID NO: 7 reported as the chain of patritumab, and U.S. Patent No. 9,249,230 and 7,705,130 and International Publication No. WO2007077028.

Exemplary HER3 antibodies also include the HER3 antibody MM-121 and any of the HER3 antibodies disclosed in US Pat. No. 7,846,440 and International Publication No. WO2008100624. Exemplary HER3 antibodies also include the EGFR/HER3 bispecific antibody DL1 and any of the monospecific or multispecific HER3 antibodies disclosed in US Pat. Nos. 9,327,035 and 8,597,652, US Publication No. 20140193414, and International Publication No. WO2010108127. .

Exemplary HER3 antibodies also include the HER2/HER3 bispecific antibody MM-111 and any of the monospecific or multispecific HER3 antibodies disclosed in US Publication Nos. 20130183311 and 20090246206 and International Publication Nos. W02006091209 and WO2005117973.

According to certain aspects, the HER3 targeting agent is patritumab from Daiichi Sankyo, cerivantumab (MM-121) from Merrimac Pharmaceuticals, rumletuzumab from Roche, elgemtumab from Novartis, Glaxosmith GSK2849330 from Klein, CDX-3379 from Celdex Therapeutics, EV20 and MP-RM-1 from Medipharma, Baresetamab (ISU104) from Isu Abxis Company, HMBD- from Hummingbird Biosciences Pte. 001 (10D1F), REGN1400 from Regeneron Pharmaceuticals, and/or anti-HER3 antibodies that bind to epitopes of HER3 recognized by AV-203 from Aveo Oncology. According to certain aspects, the anti-HER3 antibody comprises patritumab, cerivantumab or an antibody comprising the heavy chain sequence SEQ ID NO: 108 and/or light chain sequence SEQ ID NO: 109 as reported for cerivantumab, rumletuzumab or antibody comprising heavy chain sequence SEQ ID NO: 110 and/or light chain sequence SEQ ID NO: 111 as reported for rumletuzumab, heavy chain sequence SEQ ID NO: 112 and/or light chain as reported for elgemtumab or elgemtumab An antibody comprising sequence SEQ ID NO: 113, AV-203, CDX-3379, GSK2849330, EV20, MP-RM-1, ISU104, HMBD-001 (10D1F) and REGN1400. Exemplary antibodies are also listed in Table 1 along with exemplary treatment indications.

<Table 1>

Wherever specific antibodies, specific antibody heavy chains and specific antibody light chains to HER3 or to any target are disclosed in this disclosure, they are considered to be disclosed for embodiment or use in the various aspects of the present invention. It is to be understood that also intended are antibodies such as, but not limited to, immunoglobulins, such as, but not limited to, IgG, including: (i) a antibody comprising a heavy chain variable region of a disclosed antibody or heavy chain; (ii) comprising one, two or three heavy chain CDRs (e.g., by Kabat definition) of a disclosed antibody or heavy chain, (iii) comprising a light chain variable region of a disclosed antibody or light chain. and/or (iv) comprising one, two or three light chain CDRs (eg, according to the Kabat definition) of a disclosed antibody or light chain. Also, wherever an antibody heavy chain or antibody light chain comprising an N-terminal leader sequence is disclosed in this disclosure, it is also intended to be disclosed for embodiments or use in various aspects of the invention, that the leader sequence It should be understood that the corresponding heavy chain and the corresponding light chain are missing.

Additionally, the present invention provides one or more isomeric amino acid replacements having the correct mass (e.g., in any of SEQ ID NOs: 1-14 listed in Figures 1 and 2), such as replacing Ile with Leu or vice versa. Modified versions of any of the listed amino acid sequences are provided, where possible. Additionally, certain portions of these sequences may be substituted, such as by relevant portions from human immunoglobulins, to form chimeric immunoglobulins (ie, chimeric or humanized HER3). Exemplary substitutions include all or part of a human leader sequence and/or conserved regions from a human IgG1, IgG2 or IgG4 heavy chain and/or a human kappa light chain.

The sequence and structure of human HER3, human HER2, and human EGFR (HER1) are all known. The amino acid sequence of human HER3 precursor protein (receptor tyrosine-protein kinase erbB-3 isoform 1 precursor NCBI Reference Sequence: NP_001973.2) is provided herein as SEQ ID NO: 115. Various types of suitable antibodies and antibodies specific for the extracellular domain of HER3, such as human HER3, for use in various aspects of the present invention will be readily available to those skilled in the art, given known target protein amino acid sequences. It will be readily appreciated that mimics can be produced using immunization and/or panning and/or antibody engineering techniques well established in the art.

A HER3 targeting agent that is radiolabeled for use in various embodiments of the present invention can be, for example, a HER3 binding peptide, such as a chelator-containing HER3 binding peptide, such as a DOTA-containing HER3 binding peptide, such as disclosed in US Publication No. 20200121814. Any of these may be included.

According to certain aspects, the HER3 targeting agent is directed against a first epitope of HER3 and at least a second epitope of HER3, or against HER3 and one or more different antigens, such as EGFR (HER1), HER2, TROP2, and T-cell receptor gamma (TCRγ). ) multispecific targeting agents, such as multispecific antibodies, to one or more of the chain alternating reading frame proteins (TRAPs). Exemplary multispecific antibodies that may be used include bispecific antibodies to HER3/HER2, such as MM-111 from Merrimac Pharmaceuticals or MCLA-128 from Merus N.V. (ie, Xenocutuzumab); or a bispecific antibody to IGF-1R/HER3 such as MM-141 from Merrimac Pharmaceuticals (ie istiratumab); or a bispecific antibody to EGFR/HER3, such as MEHD7945A from Roche (i.e. duligotumab) or cetuximab-based bispecific or multispecific xybodies from Zingenia Inc. .

According to certain aspects, compositions comprising a mixture of a HER3 targeting agent, such as an antibody to HER3, and one or more antibodies to one or more different antigens are provided and/or used, wherein one or more of the antibodies are radiolabeled. An exemplary antibody composition comprising an antibody mixture includes at least Sym013 from Symposen with 6 monoclonal antibodies to EGFR (HER1), HER2 and HER3. In one aspect of the invention, one or more of the antibodies of Sym013, such as an anti-HER3 antibody, may be radiolabeled. A related aspect of the invention provides a composition comprising a targeting agent, such as an antibody, to EGFR (HER1), HER2, and HER3, wherein one or more or all in any combination are radiolabeled.

The present invention further relates to administration of (i) a multispecific antibody directed against two or more epitopes of HER3, or a multispecific antibody directed against an epitope of HER3 and an epitope of one or more additional different antigens, and/or (ii) administration of HER3 Treatment of a proliferative disease or disorder comprising administration of a targeting agent, such as an antibody and one or more discrete targeting agents directed against one or more cancer-associated antigens, wherein one or more of the targeting agents, such as a HER3 targeting agent, is radiolabeled Multispecific targeting agents, compositions and related methods are provided. Additional different antigens, e.g., their expression on cells involved in various diseases or disorders, such as proliferative disorders, e.g., solid tumor cancers, such as HER3, are also upregulated or can be upregulated. This may be an antigen that is upregulated. For example, additional different antigens include mesothelin, TSHR, CD19, CD123, CD22, CD30, CD45, CD171, CD138, CS-1, CLL-1, GD2, GD3, B-cell maturation antigen (BCMA), Tn Ag, prostate specific membrane antigen (PSMA), ROR1, FLT3, TROP2, T-cell receptor gamma (TCRγ) chain alternating reading frame protein (TRAP), fibroblast activation protein (FAP), calreticulin, phosphatidylserine, GRP78 (BiP), TAG72, CD38, CD44v6, CEA, EPCAM, B7H3, KIT, IL-13Ra2, interleukin-11 receptor a (IL-11Ra), PSCA, PRSS21, VEGFR2, Lewis Y, CD24, platelet-derived growth factor receptor- beta (PDGFR-beta), SSEA-4, CD20, folate receptor alpha (FRa), ERBB2 (Her2/neu), MUC1, epidermal growth factor receptor (EGFR), EGFRvIII, NCAM, protase, PAP, ELF2M, F Lin B2, IGF-I receptor, CAIX, LMP2, gp100, bcr-abl, tyrosinase, EphA2, fucosyl GM1, sLe, GM3, DR5, 5T4, TGS5, HMWMAA, o-acetyl-GD2, folate receptor beta, TEM1/ CD248, TEM7R, CLDN6, GPRC5D, CXORF61, CD97, CD 179a, ALK, polysialic acid, PLAC1, GloboH, NY-BR-1, UPK2, HAVCR1, ADRB3, PANX3, GPR20, LY6K, OR51E2, TARP, WT1, NY- ESO-1, LAGE-1a, MAGE-A1, legumain, HPV E6,E7, MAGE A1, MAGEA3, MAGEA3/A6, ETV6-AML, sperm protein 17, XAGE1, Tie 2, MAD-CT-1, MAD -CT-2, Fos-associated antigen 1, prostein, subvivin and telomerase, PCTA-1/galectin 8, KRAS, MelanA/MART1, Ras mutant, hTERT, sarcoma potential breakpoint, ML-IAP, ERG (TMPRSS2 ETS fusion gene), NA17, PAX3, androgen receptor, cyclin B1, MYCN, RhoC, TRP-2, CYP1B 1, BORIS, SART3, PAX5, OY-TES 1, LCK, AKAP-4, SSX2, RAGE- 1, human telomerase reverse transcriptase, RU1, RU2, intestinal carboxyl esterase, mut hsp70-2, CD79a, CD79b, CD72, LAIR1, FCAR, LILRA2, CD300LF, CLEC12A, BST2, EMR2, LY75, GPC3, FCRL5 , GPA7 and IGLL1.

Exemplary DR5 (death receptor 5) targeting agents that may be radiolabeled, unlabeled or drug-conjugated for use in the present invention include the monoclonal anti-DR5 antibodies mapatumumab, conatumumab, lexatumumab, Tigatuzumab, Drozitumab, and LBY-135. Such DR5 targeting agents can be radiolabeled to treat, for example, ovarian, breast, cervical, prostate, gastric, bladder, lung, melanoma, colorectal and squamous cell carcinoma cancers and any of the cancers disclosed herein. It can be used in combination with a HER3 targeting agent.

Exemplary 5T4 (trophoblast glycoprotein (TBPG)) targeting agents that may be radiolabeled, drug-conjugated or unlabeled for use in the present invention are anti-5T4 monoclonal antibodies MED10641, ALG.APV-527, Tb535 , H6-DM5, and ZV0508 as well as naptumomab estafenatox or the Fab portion thereof. Such 5T4 targeting agents are radioactive to treat, for example, ovarian, head and neck, breast, prostate, gastric, bladder, lung, melanoma, colorectal and squamous cell carcinoma cancers and any of the cancers disclosed herein. It can be used in combination with labeled HER3 targeting agents.

Exemplary HER2 (ERBB2) targeting agents that may be radiolabeled, drug-conjugated or unlabeled for use in the present invention include the monoclonal antibodies Trastuzumab and Pertuzumab. Applicants successfully conjugated trastuzumab with p-SCN-DOTA and radiolabeled the composition with ²²⁵ Ac or ¹⁷⁷ Lu. An exemplary HER2 targeting ADC that may be used is pharm-trastuzumab deluxtecan-nxki [Enhertu®; AstraZeneca/Daichi Sankyo] and trastuzumab emtansine [Roche/Genentech]. Anti-HER2 antibodies, for example, also include multispecific antibodies against any available epitope of HER3/HER2, such as bispecific antibodies such as MM-111 and MM-141/ from Merrimac Pharmaceuticals. Istiratumab, MCLA-128 from Merus NV, and MEHD7945A/duligotumab from Genentech. HER2 targeting agents are, for example, HER2-expressing cancers such as ovarian cancer, breast cancer, metastatic breast cancer, esophageal cancer, lung cancer, cervical cancer and endometrial cancer (including but not limited to both HER2- and HER3-positive) It can be used in combination with a radiolabeled HER3 targeting agent in the treatment of .

The amino acid sequences of the heavy and light chains of Trastuzumab reported by Drugbank Online are heavy chains (SEQ ID NO: 102) and light chains (SEQ ID NO: 103), and a HER2-binding antibody comprising one or both of these chains is It can be implemented or used in various embodiments of the present invention.

The amino acid sequences of the heavy and light chains of Pertuzumab reported by Drugbank Online are heavy chains (SEQ ID NO: 104) and light chains (SEQ ID NO: 105), and a HER2-binding antibody comprising one or both of these chains is It can be implemented or used in various embodiments of the present invention.

Exemplary CD33 targeting agents that may be radiolabeled, drug-conjugated, or unlabeled for use in the present invention include the monoclonal antibodies lintuzumab, gemtuzumab, and badastuximab. In combination with a radiolabeled HER3 targeting agent as disclosed herein, a CD33 targeting therapeutic can, for example, by depleting bone marrow derived suppressor cells (MDSC), for example, in solid cancers such as ovarian cancer, breast cancer, cervical cancer prostate, gastric, bladder, lung, melanoma, colorectal and squamous cell carcinoma cancers and any of the cancers disclosed herein. In one aspect, the CD33 targeting agent used in combination with the radiolabeled HER3 targeting agent is 225Ac-lintuzumab. In another aspect, the CD33 targeting agent used in combination with the radiolabeled HER3 targeting agent is the ADC gemtuzumab ozogamicin [Mylotarg®; Pfizer].

Exemplary CD38 targeting agents that may be radiolabeled, drug-conjugated, or unlabeled for use in the present invention are anti-CD38 monoclonal antibodies such as daratumumab [Darzalex®; Johnson and Johnson] and isatuximab [Sarclisa®; Sanofi] or an antigen-binding fragment thereof. Such CD38 targeting agents can, for example, treat solid tumors that can be infiltrated with CD38-positive suppressor immune cells, such as ovarian cancer, breast cancer, cervical prostate cancer, gastric cancer, bladder cancer, lung cancer, melanoma, colorectal cancer and squamous cell carcinoma. It can be used in combination with a radiolabeled HER3 targeting agent in the treatment of cancer and any of the cancers disclosed herein.

Exemplary different antigens (relative to HER3) that can be targeted by multispecific antibodies according to aspects of the present invention include at least HER1 (EGFR), HER2 and IGF-1R. Exemplary HER3 multispecific targeting agents include multispecific antibodies such as MM-111 from Merrimac Pharmaceuticals or MCLA-128 from Merus NV (ie, Xenocutuzumab); or a multispecific antibody to IGF-1R/HER3 such as MM-141 from Merrimac Pharmaceuticals (ie istiratumab); or multispecific antibodies against EGFR/HER3, such as MEHD7945A from Roche (i.e., duligotuzumab), cetuximab-based bispecific xybodies from Zingenia Inc., and multispecific antibodies from Symposen. Contains the composition Sym-013. See also Table 2 for further explanation and exemplary indications.

<Table 2>

The invention also provides a method of treating a proliferative disease or disorder comprising administering a first antibody to at least one epitope of HER3 and administering a second antibody, wherein the second antibody is different from the first antibody. It is an antibody against an epitope of HER3, or an antibody against a different antigen, such as an epitope of one or more antigens selected from the list of different antigens set forth above. One or more of the HER3 antibodies may be radiolabeled. Antibodies to different antigens may also be radiolabeled, for example in any combination.

Such combinations, presented as multispecific antibodies or more than one monoclonal antibody as indicated above, can deliver a synergistic therapeutic effect comparable to the effectiveness of monotherapy using only antibodies to HER3 while reducing the side effects of monotherapy. Moreover, such combinations may deliver improved efficacy over monotherapy, as measured by, for example, reduction in total tumor cell number, increased time to recurrence, and other indicators of patient health.

Where the method involves administration of a multispecific antibody, the first target recognition component may comprise, for example, a first full-length heavy chain and a first full-length light chain, a first Fab fragment, a first single chain variable fragment (scFv), or other types of antibodies. The second target recognition component may include, for example, a second full-length heavy chain and a second full-length light chain, a second Fab fragment, or a second single chain variable fragment (scFv) or one of the other types of antibodies. there is. Moreover, the second target recognition component may be derived from a different epitope of the HER3 antigen or may be derived from any of the antigens listed above.

The HER3 targeting agent may include a radioisotope, and any additional antibodies to other antigens may optionally include a radioisotope. According to certain aspects of the invention, where the immunotherapy comprises a bispecific antibody, one or both of the first target recognition component and the second target recognition component, or any portion of the bispecific targeting component, is a radioisotope may contain elements.

According to certain aspects of the invention, a radiolabeled targeting agent may be immunoreactive against essentially the same antigen as a control targeting agent, wherein the control targeting agent is immunoreactive against the same antigen (i.e., HER3) as the radiolabeled targeting agent. Naked or otherwise unlabeled targeting agents to the epitope.

According to certain aspects of the invention, the targeting agent may be labeled with ²²⁵ Ac and may be at least 5-fold more effective at inducing cell death of HER3-positive cells than a control monoclonal antibody, wherein the control monoclonal antibody Antibodies include naked or unlabeled antibodies directed against the same antigenic epitope as the ²²⁵ Ac labeled antibody. For example, ^{a 225} Ac labeled monoclonal antibody is at least 10-fold more effective, at least 20-fold more effective, at least 50-fold more effective at inducing apoptosis of HER3-positive cells than a control monoclonal antibody, or , or at least 100 times more effective.

According to certain aspects of the invention, the methods may include administration of labeled and unlabeled (eg, “naked”) fractions of a HER3 targeting agent, such as an antibody, antibody fragment, etc. For example, the unlabeled fraction may contain the same antibodies to the same epitope as the labeled fraction. In this way, the total radioactivity of the antibody can be varied or held constant, while the total antibody protein concentration can be held constant or varied, respectively. For example, the total protein concentration of the unlabeled antibody fraction to be administered depends on the precise nature of the disease being treated, the age and weight of the patient, the identity of the monoclonal antibody, and the marker selected for labeling the monoclonal antibody (e.g. e.g. radionuclides).

According to certain aspects of the invention, the effective amount of the anti-HER3 antibody is the maximum tolerated dose (MTD) of the anti-HER3 antibody.

According to certain method aspects of the invention, when more than one antibody is administered, these antibodies may be administered simultaneously. As such, according to certain aspects of the invention, the antibody may be provided in a single composition. Alternatively, the two antibodies can be administered sequentially. As such, the radiolabeled HER3 targeting agent can be administered before administration of the second antibody, after administration of the second antibody, or both before and after administration of the second antibody. Moreover, the second antibody can be administered prior to administration of the radiolabeled HER3-targeting agent, after administration of the radiolabeled HER3-targeting agent, or both before and after administration of the radiolabeled HER3-targeting agent.

According to certain aspects of the methods of the invention, the radiolabeled HER3 targeting agent is administered every 7 days, 10 days, 12 days, 14 days, 20 days, 24 days, 28 days, 35 days, and 42 days throughout the treatment period. It may be administered according to an administration schedule selected from the group consisting of one administration, wherein the treatment period includes at least two administrations.

According to certain aspects of the invention, the radiolabeled HER3 targeting agent is administered in two doses, e.g., on days 1 and 5, 6, 7, 8, 9, or 10 of the treatment period. administration, or according to an administration schedule including administration on days 1 and 8 of the treatment period.

Administration of the radiolabeled HER3 targeting agent of the present invention can be provided in a number of ways depending on the area to be treated and whether local or systemic treatment is desired, in addition to other therapeutic agents. Administration can be intratracheal, intranasal, epidermal and transdermal, oral or parenteral. Parenteral administration includes intravenous, intraarterial, subcutaneous, intraperitoneal or intramuscular injection or infusion; or intracranial, eg intrathecal or intraventricular administration. In some embodiments, a sustained release formulation comprising the targeting agent(s) and/or other therapeutic agent may be administered. The various agents may be administered as a single treatment or as a series of treatments continued as needed for a period of time to reduce or alleviate one or more symptoms of cancer or achieve another desired effect.

The dose(s) may vary depending, for example, on the identity, size and condition of the subject, further depending on the route by which the composition is administered and the effect desired. Appropriate doses of therapeutic agents depend on the potency with respect to expression or activity to be modulated. A therapeutic agent may be initially administered to an animal (eg, human) at a relatively low dose, with the dose subsequently increased until an adequate response is obtained.

The radiolabeled HER3 targeting agent may be administered concurrently or sequentially with one or more additional therapeutic agents. Moreover, when more than one additional therapeutic agent is included, the additional therapeutic agents can be administered simultaneously or sequentially with each other and/or with the radiolabeled HER3 targeting agent.

HER3 targeting agent radiolabeling thing

The HER3 targeting agents and other targeting agents disclosed herein can be used to target a radioisotope, such as a beta emitter (e.g., ¹⁷⁷ Lu), for example, via conjugation to a chelator molecule, and via chelation of a radioisotope thereto. or with an alpha emitter (eg ²²⁵ Ac). According to certain aspects, the targeting agent is an asparagine- in the constant region, such as the heavy chain CH2 domain, for the purpose of finding a unique conjugation site, glutamine (i.e., Gln-295, Q295) so that it can conjugate with the bifunctional chelator molecule. 297 (Asn-297, N297; Kabat number).

According to certain aspects, the radiotherapeutic agent may be an antibody that may have reduced disulfide bonds, such as with a reducing agent, which may then be converted to dehydroalanine for conjugation with a bifunctional chelator molecule.

According to certain aspects, the radiotherapeutic agent may be an antibody whose disulfide bonds have been reduced using a reducing agent, which is then conjugated to a bifunctional chelator molecule via an aryl bridge. For example, according to certain aspects, a linker molecule such as 3,5-bis(bromomethyl)benzene may be used to bridge free sulfhydryl groups on an antibody.

According to certain aspects, the radiotherapeutic agent can be an antibody in which certain specific pre-existing amino acids can be replaced with cysteine(s) and then used for site-specific labeling.

Exemplary chelators that can be linked to targeting agents in various aspects of the invention include: 1,4,7,10-tetraazacyclododecane-1,4,7-triacetic acid (DO3A) or a derivative thereof ; 1,4,7-triazacyclononane-1,4-diacetic acid (NODA) or a derivative thereof; 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA) or a derivative thereof; 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA) or a derivative thereof; 1,4,7-triazacyclononane, 1-glutaric acid-4,7-diacetic acid (NODAGA) or a derivative thereof; 1,4,7,10-tetraazacyclodecane, 1-glutaric acid-4,7,10-triacetic acid (DOTAGA) or derivatives thereof; 1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid (TETA) or a derivative thereof; 1,4,8,11-tetraazabicyclo[6.6.2]hexadecane-4,11-diacetic acid (CB-TE2A) or a derivative thereof; diethylene triamine pentaacetic acid (DTPA), a diester thereof or a derivative thereof; 2-cyclohexyl diethylene triamine pentaacetic acid (CHX-A″-DTPA) or a derivative thereof; deforoxamine (DFO) or a derivative thereof; 1,2-[[6-carboxypyridin-2-yl]methylamino] ethane (H ₂ dedpa) or a derivative thereof DADA or a derivative thereof 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetra(methylene phosphonic acid) (DOTP) or a derivative thereof; 4-amino-6-[[16-[(6-carboxypyridin-2-yl)methyl]-1,4,10,13-tetraoxa-7,16-diazacyclooctadec-7-yl]methyl ]Pyridine-2-carboxylic acid (MACROPA-NH ₂ ) or a derivative thereof; MACROPA or a derivative thereof; 1,4,7,10-tetrakis(carbamoylmethyl)-1,4,7,10-tetraaza Cyclododecane (TCMC) or its derivatives {4-[2-(bis-carboxymethylamino)-ethyl]-7-carboxymethyl-[1,4,7]triazonan-1-yl}-acetic acid (NETA ) or a derivative thereof; Diamsar or a derivative thereof; 1,4,7-triazacyclononane-1,4,7-tris[methyl(2-carboxyethyl)phosphinic acid (TRAP, PRP9, TRAP-Pr) or its N,N'-bis(6-carboxy-2-pyridylmethyl)ethylenediamine-N,N'-diacetic acid (H4octapa) or its derivatives;N,N'-[1-benzyl-1,2, 3-triazol-4-yl]methyl-N,N'-[6-(carboxy)pyridin-2-yl]-1,2-diaminoethane (H2azapa) or a derivative thereof; N,N"-[[ 6-(carboxy)pyridin-2-yl]methyl]diethylenetriamine-N,N',N"-triacetic acid (H5decapa) or a derivative thereof; N,N'-bis(2-hydroxy-5-sulfo Benzyl)ethylenediamine-N,N'-diacetic acid (SHBED) or a derivative thereof N,N'-bis(2-hydroxybenzyl)ethylenediamine-N,N'-diacetic acid (HBED) or a derivative thereof 3 ,6,9,15-tetraazabicyclo[9.3.1]pentadeca-1(15),11,13-triene-3,6,9,-triacetic acid (PCTA) or its derivatives; B (DFO) or a derivative thereof: N,N'-(methylenephosphonate)-N,N'-[6-(methoxycarbonyl)pyridin-2-yl]methyl-1,2-diaminoethane ( H6phospa) or a derivative thereof; 1,4,7,10,13,16-hexaazacyclohexadecane-N,N',N",N'",N"",N""'-hexaacetic acid (HEHA) or a derivative thereof; 1,4,7,10,13-pentaazacyclopentadecane-N,N',N",N'",N""-pentaacetic acid (PEPA) or a derivative thereof; or 3,4,3-LI(1,2-HOPO) or a derivative thereof.

According to certain aspects, the targeting agent may be radiolabeled via chemical conjugation of a suitable bifunctional chelator capable of chelating one or more radionuclides. Exemplary chelator molecules that may be used are p-SCN-Bn-DOTA, NH ₂ -DOTA, NH ₂ -(CH ₂ ) _1-20 -DOTA, NH ₂ -(PEG) _1-20 -DOTA, HS-DOTA , HS-(CH ₂ ) _1-20 -DOTA, HS-(PEG) _1-20 -DOTA, dibromo-S-(CH ₂ ) _1-20 -DOTA, dibromo-S-(PEG) _{1 -20} -DOTA, p-SCN-Bn-DOTP, NH ₂ -DOTP, NH ₂ -(CH ₂ ) _1-20 -DOTP, NH ₂ -(PEG) _1-20 -DOTP, HS-DOTP, HS-( CH ₂ ) _1-20 -DOTP, HS-(PEG) _1-20 -DOTP, dibromo-S-(CH ₂ ) _1-20 -DOTP, and dibromo-S-(PEG) _1-20 - Includes DOTP.

For example, a chelator molecule can be attached to a targeting agent through a linker molecule. Exemplary linker molecules include:

-CH ₂ (C ₆ H ₄ )NH ₂ or -CH ₂ (C ₆ H ₄ )NH-XY;

where X is

-R ₂ -CH ₂ CH ₂ O(CH ₂ CH ₂ O) _n CH ₂ CH ₂ -,

-R ₂ -CH ₂ CH ₂ NHC(O)CH ₂ CH ₂ O(CH ₂ CH ₂ O) _n CH ₂ CH ₂ -,

-R ₂ -(CH ₂ ) _n CH ₂ -,

-R ₂ -CH ₂ CH ₂ NHC(O)(CH ₂ ) _n CH ₂ -,

-R ₂ -CH(C(O)R ₃ )CH ₂ -, where R ₃ is -OH or a short peptide (1-20 amino acids);

-R ₂ -CH ₂ CH ₂ O(CH ₂ CH ₂ O) _n CH ₂ C(O)O-, or

-R ₂ -CH ₂ CH ₂ NHC(O)CH ₂ CH ₂ O(CH ₂ CH ₂ O) _n CH ₂ CC(O)O-;

where n is 1-20,

R ₂ is -C(O)- or -C(S)NH-;

Y is -NH ₂ or -SR ₄ -, wherein R ₄ is -H or -CH ₂ -3,5-bis(bromomethyl)benzene.

Targeting agents, such as protein targeting agents such as antibodies and antigen-binding antibody fragments, and peptide targeting agents can be conjugated with chelators to radiolabel the targeting agent, for example through chelation of a radionuclide. For example, such a protein or peptide targeting agent comprising lysine(s) is “p-SCN-Bn-DOTA” [catalog # B205; Macrocyclics, Inc. (Macrocyclics, Inc.; Plano, Texas, USA)], a bifunctional agent also known as S-2-(4-isothiocyanatobenzyl)-1,4,7,10-tetraazacyclododecane tetraacetic acid. Can be conveniently conjugated to the DOTA chelating moiety using p-SCN-Bn-DOTA can be synthesized by multi-step organic synthesis fully described in U.S. Patent No. 4,923,985. Chelation of the radionuclide by the DOTA moiety can be performed before and/or after chemical conjugation of the antibody with p-SCN-Bn-DOTA.

A method for labeling chelator-conjugated targeting agents with exemplary radionuclides is described in Example 1.

diagnostic aspect

The methods disclosed herein may include diagnosing a subject to determine whether HER3-positive cells are present, the degree to which they are present, and/or their localization. HER3-positive cells can be present in many biological specimens, such as circulating cells in a blood sample from a subject or tumor cells in a biopsy of a subject. In one aspect, the diagnosing step may generally include obtaining a blood or tissue sample from a subject and immobilizing the sample on a substrate. The presence or absence of the HER3 antigen can be detected using a diagnostic antibody, peptide or small molecule, wherein the diagnostic antibody, peptide or small molecule is labeled with any of the standard imaging labels known in the art. Exemplary labeling agents include, for example, radiolabels such as ³ H, ¹⁴ C, ³² P, ³⁵ S, and ¹²⁵ I; fluorescent or chemiluminescent compounds such as fluorescein isothiocyanate, rhodamine or luciferin; and enzymes such as alkaline phosphatase, β-galactosidase or horseradish peroxidase. Exemplary HER3 targeting agents for use in such diagnostic assays include human or humanized antibodies to HER3.

In another aspect, the method is a radionuclide, such as any of ¹⁸ F, ¹¹ C, ⁶⁸ Ga, ⁶⁴ Cu, ⁸⁹ Zr, or ¹²⁴ I for PET imaging, or ^99m Tc or ¹¹¹ for SPECT imaging. diagnosing the subject to determine if HER3-positive cells are present using a HER3 targeting agent labeled with In. Thus, the method comprises administering to a subject a HER3 targeting agent labeled with one or more of ¹⁸ F, ¹¹ C, ⁶⁸ Ga, ⁶⁴ Cu, ⁸⁹ Zr, ¹²⁴ I, ^99m Tc, or ¹¹¹ In, and - performing an invasive imaging technique, such as performing a PET or SPECT scan on the subject. The method can include administering a radiolabeled HER3 targeting agent to a subject for imaging, and performing imaging after a sufficient time for the targeting agent to bind to a target in a tissue of the subject. The amount of time sufficient for a targeting agent to bind to a target in a tissue of a subject can be, for example, at least 20 minutes, at least 30 minutes, at least 60 minutes, or any number or subrange of minutes within the range of 20 minutes to 360 minutes. there is. According to one particular aspect of the method, the radiolabeled HER3 targeting agent may comprise ⁶⁸ Ga, ⁸⁹ Zr, or ¹¹¹ In, and any of the methods disclosed herein (eg, as described in Example 1) can be labeled using

If the subject has, for example, HER3-positive cancer cells above a predetermined or pre-selected threshold level, or other indications of HER3-positive cancer/tumor, the treatment methods of the invention disclosed by this application can be performed, That is, administration of a therapeutically effective amount of a radiolabeled HER3 targeting agent alone or in combination with one or more additional therapeutic agents can be performed.

Additional Therapeutics and Treatment Modalities

Methods of the invention comprising administering a radiolabeled HER3 targeting agent therapeutic alone or in combination with other targeting agents may further include administration of additional therapeutic agents or therapeutic modalities. According to certain aspects, the additional agent may be associated with a disease or condition being treated by the radiolabeled HER3 targeting agent. Such administration may be concurrent with, separate from, or sequential with administration of an effective amount of the HER3 targeting agent. For simultaneous administration, the agents may be administered as one composition or as separate compositions, as appropriate.

Exemplary additional therapeutic agents and treatment modalities that can be used in combination or in conjunction with radiolabeled HER3 targeting agents include at least chemotherapeutic agents, small molecule oncology drugs, anti-inflammatory agents, immunosuppressive agents, immunomodulatory agents, immune checkpoint therapies, DDR inhibitors, CD47 blockers, external beam radiation, brachytherapy, or any combination thereof. Exemplary additional agents and treatment modalities that can be used in combination or in conjunction with radiolabeled HER3 targeting agents, either alone or in combination with other targeting agents as disclosed herein, are described further below.

A. Chemotherapeutic agents and others small molecule agent

Exemplary chemotherapeutic agents include nitrogen mustards such as alkylating agents including mechlorethamine, cyclophosphamide, ifosfamide, melphalan and chlorambucil; nitrosoureas such as carmustine (BCNU), lomustine (CCNU) and semustine (methyl-CCNU); Temodal™ (temozolomide), ethylenimine/methylmelamine such as triethylenemelamine (TEM), triethylene, thiophosphoramide (thiotepa), hexamethylmelamine (HMM, altretamine); alkyl sulfonates such as busulfan; triazines such as dacarbazine (DTIC); folic acid analogs such as methotrexate and trimetrexate, pyrimidine analogs such as 5-fluorouracil (5FU), fluorodeoxyuridine, gemcitabine, cytosine arabinoside (AraC, cytarabine), 5- Azacytidine, 2,2'-difluorodeoxycytidine, purine analogs such as 6-mercaptopurine, 6-thioguanine, azathioprine, T-deoxycoformycin (pentostatin), erythrohydride antimetabolites including oxynonyladenine (EHNA), fludarabine phosphate and 2-chlorodeoxyadenosine (cladribine, 2-CdA); antimitotic drugs such as paclitaxel, vinblastine (VLB), vinca alkaloids including vincristine and vinorelbine, natural products including taxotere, estramustine and estramustine phosphate; pipodophyllotoxins such as etoposide and teniposide; antibiotics such as actinomycin D, daunomycin (rubidomycin), doxorubicin, mitoxantrone, idarubicin, bleomycin, plicamycin (mitramycin), mitomycin C, and actinomycin; enzymes such as L-asparaginase; biological response modifiers such as interferon-alpha, IL-2, G-CSF and GM-CSF; platinum coordination complexes such as oxaliplatin, cisplatin and carboplatin, anthracenediones such as mitoxantrone, methylhydrazine derivatives including substituted ureas such as hydroxyurea, N-methylhydrazine (MIH) and procarbazine, adrenocortical inhibitors other agents, including mitotane (o, p-DDD) and aminoglutethimide; antagonists including hormone and corticosteroid antagonists such as prednisone and equivalents, dexamethasone and aminoglutethimide; Gemzar™ (gemcitabine), progestins such as hydroxyprogesterone caproate, medroxyprogesterone acetate and megestrol acetate; estrogens such as diethylstilbestrol and ethinyl estradiol equivalents; antiestrogens such as tamoxifen; androgens including testosterone propionate and fluoxymesterone/equivalents; anti-androgens such as flutamide, gonadotropin-releasing hormone analogues and leuprolide; and anti-neoplastic agents including non-steroidal anti-androgens such as flutamide.

(i) histone deacetylase (HDAC) inhibitors such as vorinostat (suberoylanilide hydroxamic acid; SAHA), romidepsin, belinostat (PDX101), panobinostat (LBH589) and tucidinostat, methylating agents (eg, Vidaza); (ii) LSD1 inhibitors such as seclidemstat, TCP (tranilcypromine), ORY-1001 (iadademstat), GSK2879552 (GSK), INCB059872 (Imago BioSciences), IMG-7289 (Bomedemstat; Imago Biosciences), ORY-2001 (Bapidemstat), and CC-90011 (Celgene); and (iii) therapies targeting epigenetic mechanisms, including but not limited to release of transcriptional repression (ATRA) therapy, can also be performed using radiolabeled HER3-targeting agents and/or other radiolabeled targets as disclosed herein. It can be used in combination or in conjunction with the topic and combinations thereof.

According to certain aspects of the invention, the chemotherapeutic agent includes at least a radiosensitizer such as temozolomide, cisplatin and/or fluorouracil.

Additional agents include, for example, bcl-2 inhibitors such as navitoclax or venetoclax [Venclexta®; Abbvie], and the combination can be used, for example, in the treatment of solid tumors such as breast and lung cancers such as small cell lung carcinoma (SCLC).

Additional agents include, for example, cyclin dependent kinase CDK4 and CDK6 inhibitors such as palbociclib [Ibrance®; Pfizer], and the combination can be used, for example, with or without an aromatase inhibitor, for the treatment of solid cancers, such as breast cancer, such as HR-positive and HER2-negative breast cancer.

Additional agents include, for example, erlotinib [Tarceva®; Roche], the combination being useful, for example, in the treatment of solid tumor cancers such as non-small cell lung cancer (NSCLC), eg, those having mutations in the epidermal growth factor receptor (EGFR), and pancreatic cancer. can be used

Additional agents include, for example, sirolimus or everolimus [Affinitor®; Novartis], and the combination can be used, for example, in the treatment of solid tumor cancers such as melanoma and breast cancer.

Additional agents include, for example, pemetrexed [Alimta®; Eli Lilly], and the combination can be used, for example, in the treatment of solid cancers such as mesothelioma, such as pleural mesothelioma and lung cancer, such as non-small cell lung cancer (NSCLC).

The additional therapeutic agent may be administered according to any standard dosing regimen known in the art, for example. For example, a therapeutic agent can be administered at a concentration ranging from 1 to 500 mg/m ² , the amount calculated as a function of patient surface area (m ² ). For example, an exemplary dose of the chemotherapeutic agent paclitaxel may include 15 mg/m ² to 275 mg/m ² , and an exemplary dose of docetaxel may include 60 mg/m ² to 100 mg/m ² An exemplary dose of epitilone may include 10 mg/m ² to 20 mg/m ² , and an exemplary dose of calicheamicin may include 1 mg/m ² to 10 mg/m ² there is. Although exemplary dosages are listed herein, they are provided for reference only and are not intended to limit the dosage ranges of the agents of the invention disclosed herein.

B. External beam radiation and/or brachytherapy

An additional treatment modality administered in conjunction with a HER3 targeting agent, and optionally any other of the additional therapeutic agents disclosed herein, may be ionizing radiation, such as administered via external beam radiation or brachytherapy. Such radiation is usually performed by a machine placed outside the patient's body using X-rays, gamma rays, or charged particles (eg, protons or electrons) (external beam radiation therapy) or from a source (external beam radiation therapy) placed inside the patient's body. refers to producing ionizing radiation, such as delivered by internal radiation therapy or brachytherapy).

External beam radiation or brachytherapy can enhance the targeted radiation injury delivered by a radiolabeled HER3-targeting agent, and thus can be administered sequentially with a HER3-targeting agent, such as before and/or after administration of the HER3-targeting agent, or as a HER3-targeting agent. It can be delivered simultaneously with administration of the targeting agent.

External beam radiation or brachytherapy can be planned and administered in conjunction with imaging-based techniques such as computed tomography (CT) and/or magnetic resonance imaging (MRI) to accurately determine the dose and location of the radiation to be administered. For example, patients treated with any of the radiolabeled HER3 targeting agents disclosed herein may be imaged using either CT or MRI to determine the dose and location of radiation to be administered by external beam radiation or brachytherapy. It can be.

In various embodiments, the radiation therapy is whole body radiation therapy, conventional external beam radiation therapy, stereotactic radiosurgery, stereotactic body radiation therapy, 3-D stereoscopic radiation therapy, intensity modulated radiation therapy, image-guided radiation therapy, tomography, and It may be selected from the group consisting of brachytherapy. According to certain aspects, radiation therapy can be given as a single dose or as divided doses, eg in two or more fractions. For example, doses can be administered such that each fraction contains 2-20 Gy (eg, a radiation dose of 50 Gy can be divided into 10 fractions containing 5 Gy each). Two or more fractions are consecutively or sequentially, such as once every 2 days, once every 3 days, once every 4 days, once every 5 days, once every 6 days, once every 7 days, or combinations thereof. may be administered.

C. Immune Checkpoint Therapy

The additional agent(s) administered in conjunction with the HER3 targeting agent may be an immune checkpoint therapy. Cancer cells have developed means to circumvent the immune system's standard checkpoints. For example, cancer cells can reduce the expression of tumor antigens, reduce tumor antigen presentation due to downregulation of MHC class I and II molecules, secrete immunosuppressive cytokines such as TGFb, immunosuppressive cells such as regulatory T cells (Tregs) or through recruitment or induction of bone marrow-derived suppressor cells (MDSC) and overexpression of specific ligands [eg, programmed death ligand-1 (PD-L1)] that suppress the host's pre-existing antitumor immunity. turned out to be avoided.

Another major mechanism of immune suppression by cancer cells is a process known as “T-cell depletion” due to chronic exposure to tumor antigens and is characterized by upregulation of inhibitory receptors. These inhibitory receptors serve as immune checkpoints to prevent uncontrolled immune responses.

PD-1 (ie programmed cell death protein 1) and its ligands PD-L1 and PD-L2, CTLA-4 (ie cytotoxic T-lymphocyte associated protein-4) and its ligands CD80 and CD86, LAG3 (i.e. lymphocyte-activating gene 3), B and T lymphocyte attenuator, TIGIT (T-cell immunoreceptor with Ig and ITIM domains), TIM-3 (i.e. T-cell immunoglobulin and mucin-domain containing protein 3) ), and VISTA (V-domain immunoglobulin inhibitor of T cell activation), various immune checkpoints that function at different levels of T cell immunity have been described in the literature.

Improving the efficacy of the immune system through therapeutic intervention is a particularly exciting development in cancer treatment. As indicated, checkpoint inhibitors such as CTLA-4 and PD-1 prevent autoimmunity and generally protect tissues from immune collateral damage. In addition, stimulation checkpoints such as OX40 (ie tumor necrosis factor receptor superfamily, member 4; TNFR-SF4), CD137 (ie TNFR-SF9), GITR (ie glucocorticoid-induced TNFR), CD27 (ie , TNFR-SF7), CD40 (ie cluster of differentiation 40) and CD28 activate and/or promote the expansion of T cells. Modulation of the immune system by inhibition or overexpression of these proteins is currently a promising area of research.

Thus, a promising therapeutic strategy is to use immune checkpoint therapies that can eliminate specific blockages to the immune system utilized by cancer cells, in combination with the HER3 targeting agents disclosed herein. For example, antibodies to certain immune checkpoint inhibitors (ICIs) can block interactions between checkpoint inhibitor proteins and their ligands, thus preventing signaling events that might otherwise suppress the immune response to tumor cells. can

Moreover, there is growing preclinical evidence supporting the ability of radiation to synergize with ICI antibodies, as evidenced by a growing number of clinical trials evaluating combinations of external beam radiation and immune checkpoint therapies across a range of tumor types and ICI antibodies. It is also being explored in the clinic (Lamichhane, 2018). Clinical evidence supporting this combination has been generated in melanoma and two studies demonstrating the clinical benefit of using radiation in combination with ipilimumab, an anti-cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) ICI antibody. There are several studies (Twyman-Saint Vistor, 2015).

Accordingly, it is an object of the invention disclosed herein to provide a therapy for the treatment of cancer using a HER3 targeting agent in combination with one or more immune checkpoint therapies, such as an ICI antibody.

Immune checkpoint therapies of the present invention include molecules that wholly or partially reduce, inhibit, interfere with or modulate one or more checkpoint proteins. Checkpoint proteins regulate T-cell activation or function. Immune checkpoint therapy can unblock pre-existing suppression of an immune response by binding to or otherwise inactivating a checkpoint inhibitor. Immune checkpoint therapy can include monoclonal antibodies, humanized antibodies, fully human antibodies, antibody fragments, small molecule therapeutics, or combinations thereof.

Exemplary immune checkpoint therapies include checkpoint proteins such as inhibitory receptors CTLA-4, PD-1, TIM-3, VISTA, BTLA, LAG-3 and TIGIT, and/or activating receptors CD28, OX40, CD40, GITR, CD137 , can specifically bind to and inhibit CD27 and HVEM. Additionally, immune checkpoint therapies include checkpoint proteins described above, such as PD-L1, PD-L2, PD-L3 and PD-L4 (ligands for PD-1); CD80 and CD86 (ligands for CTLA-4); CD137-L (ligand for CD137); and GITR-L (ligand of GITR). Other exemplary immune checkpoint therapies include checkpoint proteins such as CD226, B7-H3, B7-H4, BTLA, TIGIT, GALS, KIR, 2B4 (all NK, γδ and memory CD8+ (αβ) T expressed in cells), CD160 (also referred to as BY55), and CGEN-15049.

At the heart of the immune checkpoint process are the CD137, CTLA-4 and PD-1 immune checkpoint pathways.

The CTLA-4 and PD-1 pathways are thought to operate at different stages of the immune response. CTLA-4 is considered a “leader” of immune checkpoint inhibitors (ICIs) because it typically arrests potentially autoreactive T cells in the early stages of naive T-cell activation in the lymph node. The PD-1 pathway regulates previously activated T cells in the later stages of the immune response, primarily in peripheral tissues. Moreover, patients with advanced cancer have been shown to lack PD-L1 upregulation by tumor cells or tumor-infiltrating immune cells. Thus, immune checkpoint therapies targeting the PD-1 pathway may be particularly effective in tumors where this immunosuppressive axis is at play, and reversing the balance to an immune protective milieu can reactivate and enhance pre-existing antitumor immune responses. . PD-1 blockade can be achieved by a variety of mechanisms, including antibodies that bind to PD-1 or its ligand, PD-L1.

According to certain aspects of the invention disclosed by this disclosure, an immune checkpoint therapy may include an inhibitor of the PD-1 checkpoint, which interacts with PD-1 and one or more of its binding partners, such as PD-L1 and PD-L2. may reduce, block, inhibit, abolish or interfere with signal transduction resulting from the action. Inhibitors of the PD-1 checkpoint reduce anti-PD-1 antibodies, antigen-binding fragments, fusion proteins, oligopeptides, and signal transduction resulting from the interaction of PD-1 with PD-L1 and/or PD-L2; It may be another molecule that blocks, inhibits, abolishes or interferes. In some embodiments, a PD-1 checkpoint inhibitor reduces negative co-stimulatory signals mediated by or through cell surface proteins expressed on T lymphocytes, making dysfunctional T-cells less dysfunctional (e.g., , enhancing effector responses to antigen recognition). In some embodiments, the PD-1 checkpoint therapy is an anti-PD-1 antibody.

Thus, according to certain aspects of the invention, an immune checkpoint therapy may include a monoclonal antibody to an immune checkpoint inhibitor (ICI) such as CTLA-4, PD-1 or PD-L1.

According to certain aspects, the ICI antibody may be an antibody to PD-1. The ICI antibody may be an anti-PD-1 antibody, such as nivolumab. For example, inhibitors of PD-1 biological activity (or ligands thereof) are disclosed in US Patent No. 7,029,674. Exemplary antibodies to PD-1 include anti-mouse PD-1 antibody clone J43 from BioXcell (Cat #BE0033-2); anti-mouse PD-1 antibody clone RMP1-14 (Cat #BE0146) from BioXcell; mouse anti-PD-1 antibody clone EH12; Merck's MK-3475 anti-mouse PD-1 antibody (Keytruda®, pembrolizumab, lambrolizumab); and AnaptysBio's anti-PD-1 antibody known as ANB011; antibody MDX-1106 (ONO-4538); human IgG4 monoclonal antibody nivolumab from Bristol-Myers Squibb (Opdivo®, BMS-936558, MDX1106); AstraZeneca's AMP-514, and AMP-224; and pidilizumab (CT-011), CureTech Ltd.

According to certain aspects, the immune checkpoint therapy is an inhibitor of PD-L1. Exemplary inhibitors of PD-L1 include antibodies (eg, anti-PD-L1 antibodies, i.e., ICI antibodies), RNAi molecules (eg, anti-PD-L1 RNAi), antisense molecules (eg, anti-PD-L1 antibodies). PD-L1 antisense RNA), dominant negative proteins (eg, dominant negative PD-L1 protein) and small molecule inhibitors. Exemplary anti-PD-L1 antibodies include clone EH12. Exemplary antibodies to PD-L1 include Genentech's MPDL3280A (RG7446); anti-mouse PD-L1 antibody clone 10F.9G2 from BioXcell (Cat #BE0101); anti-PD-L1 monoclonal antibodies MDX-1105 (BMS-936559) and BMS-935559 from Bristol-Myers Squibb; MSB0010718C; mouse anti-PD-L1 clone 29E.2A3; and AstraZeneca's MEDI4736 (Durvalumab).

According to certain aspects, the immune checkpoint therapy may be an inhibitor of PD-L2 or reduce the interaction between PD-1 and PD-L2. Exemplary inhibitors of PD-L2 include antibodies (eg, anti-PD-L2 antibodies, i.e., ICI antibodies), RNAi molecules (eg, anti-PD-L2 RNAi), antisense molecules (eg, anti-PD-L2 antibodies). PD-L2 antisense RNA), dominant negative proteins (eg, dominant negative PD-L2 protein), and small molecule inhibitors. Antibodies include monoclonal antibodies, humanized antibodies, deimmunized antibodies, and Ig fusion proteins.

According to certain aspects, the immune checkpoint therapy can be an inhibitor of CTLA-4, such as an anti-CTLA-4 antibody, ie an ICI antibody. According to one aspect, the ICI antibody can be ipilimumab. Anti-CTLA-4 antibodies can block the binding of CTLA-4 to CD80 (B7-1) and/or CD86 (B7-2) expressed on antigen-presenting cells. Exemplary antibodies to CTLA-4 include Bristol-Myers Squibb's anti-CTLA-4 antibody ipilimumab (also known as Yervoy®, MDX-010, BMS-734016 and MDX-101); anti-CTLA4 antibody, clone 9H10 from Millipore; Pfizer's Tremelimumab (CP-675,206, Ticilimumab); and anti-CTLA-4 antibody clone BNI3 from Abcam. According to certain aspects, the immune checkpoint inhibitor can be a nucleic acid inhibitor of CTLA-4 expression.

CD137 (also known as “TNF receptor superfamily member 9”) is a costimulatory receptor member of the tumor necrosis factor receptor superfamily and mediates CD28-dependent and -independent T-cell costimulation (Bartkowiak, 2015). CD137 is inducibly expressed by T cells, natural killer (NK) cells, dendritic cells (DC), B cells, and other cells of the immune system. The protein consists of a 255 amino acid protein with a short N-terminal cytoplasmic portion, a transmembrane region, and an extracellular domain containing three cysteine-rich motifs. Ligation of CD137 by its ligand CD137L (4-1BBL; TNFSF9), which is primarily, but not exclusively, expressed on antigen-presenting cells (APCs), induces a variety of T cell responses, such as cell expansion, increased cytokine secretion, and activation-induction. induces the prevention of cell death. Thus, this ligation serves to activate the immune system. However, the cis-interaction between CD137 and CD137L also strongly downregulates the expression of CD137L (Kwon, 2015). Thus, CD137 ligand functions to control the extent and kinetics of CD137-mediated immune system activation (Kwon, 2015). Importantly, CD137 expressed on human NK cells is upregulated when combined with anti-tumor antibodies bound to tumor cells (Wei, 2014).

Thus, according to certain aspects of the invention disclosed herein, an immune checkpoint therapy may include an antibody to CD137, which may be used to activate the immune system, thereby in combination with a HER3 targeting agent disclosed herein. Thus, therapies for cancer can be provided. Exemplary anti-CD137 antibodies that may be used include US Publication Nos. 20140274909; 20130280265; 20130273078; 20130071403; 20120058047; 20110104049; 20110097313; 20080166336; 20080019905; 20060188439; 20060182744; 20060121030; and 20030223989.

According to certain aspects of the invention, an immune checkpoint therapy may include more than one modulator of an immune checkpoint protein. As such, an immune checkpoint therapy may include a first antibody or inhibitor against a first immune checkpoint protein and a second antibody or inhibitor against a second immune checkpoint protein.

D. DNA Damage Response Inhibitors

Additional agents administered in conjunction with the HER3 targeting agent may be one or more DNA damage response inhibitors (DDRi). DNA damage can be due to endogenous factors, such as spontaneous or enzymatic reactions, chemical reactions, or replication errors, or it can be due to exogenous factors, such as UV or ionizing radiation or genotoxic chemicals. The repair pathways that overcome this damage are collectively referred to as the DNA damage response or DDR. These signaling networks are responsible for detecting and orchestrating the cell's response to certain forms of DNA damage, most notably double-strand breaks and replication stress. After treatment with many types of DNA-damaging drugs and ionizing radiation, cells depend on DDR for survival. It has been shown that disruption of the DDR can increase the susceptibility of cancer cells to these DNA damaging agents and thus improve the response of patients to such therapy.

Within the DDR, there are several DNA repair mechanisms including base excision repair, nucleotide excision repair, mismatch repair, homologous recombination repair, and non-homologous end joining. Approximately 450 human DDR genes encode proteins that play a role in physiological processes. Dysregulation of the DDR causes a variety of disorders including genetic, neurodegenerative, immune, cardiovascular, and metabolic diseases or disorders, and cancer. For example, the genes OGG1 and XRCC1 are part of the base excision repair mechanism of the DDR, and mutations in these genes are found in renal, breast and lung cancer, whereas the genes BRCA1 and BRCA2 are involved in the homologous recombination repair mechanism and these genes Mutations in β increase the risk of breast, ovarian, prostate, pancreatic, as well as gastrointestinal and hematological cancers, and melanoma. Exemplary DDR genes are provided in Table 3 .

It is an object of the invention disclosed herein to administer radiolabeled HER3 targeting agents that deliver ionizing radiation in combination with DDRi. Thus, according to certain aspects, the additional agent(s) administered with the HER3 targeting agent can target proteins within the DDR, i.e., are DDR inhibitors or DDRi, thereby maximizing DNA damage or, for example, in G1 and S-phases. In the presence of damage in , it can inhibit repair and/or prevent repair in G2, bringing the maximum amount of DNA damage to mitosis, resulting in cell death.

<Table 3>

Moreover, one or more DDR pathways can be targeted to ensure cell death, ie lethality to targeted cancer cells. For example, mutations in the BRCA1 and 2 genes alone may not be sufficient to ensure cell death, as other pathways, such as the PARP1 base excision pathway, may play a role in repairing DNA damage. Thus, combinations of multiple DDRi inhibitors or combinations of DDRi with anti-angiogenic agents or immune checkpoint inhibitors such as those listed above are possible and are an object of the invention disclosed herein.

exemplary DDRi - ATM and ATR inhibitors

Mutated telangiectatic ataxia (ATM) and mutated and Rad-3 related telangiectatic ataxia (ATR) are members of the phosphatidylinositol 3-kinase related kinase (PIKK) family of serine/threonine protein kinases.

ATM is a serine/threonine protein kinase that is recruited and activated by DNA double-strand breaks. ATM phosphorylates several key proteins that initiate activation of DNA damage checkpoints, resulting in cell cycle arrest, DNA repair or cellular apoptosis. Several of these targets, including p53, CHK2 and H2AX, are tumor suppressors. This protein is named after telangiectatic ataxia, a disorder caused by mutations in ATM. ATM belongs to the superfamily of phosphatidylinositol 3-kinase related kinases (PIKKs), which includes six serine/threonine protein kinases that show sequence similarity to phosphatidylinositol 3-kinases (PI3Ks).

Like ATM, ATR is one of the central kinases involved in DDR. ATR is activated by single-stranded DNA structures, which can occur, for example, at excised DNA DSBs or delayed replication forks. When DNA polymerase is delayed during DNA replication, the replicating helicase continues to unwind DNA before the replication fork, creating long, single-stranded DNA (ssDNA).

ATM has been shown to benefit cancer cells by providing resistance to chemotherapeutic agents, thus helping tumor growth and survival. Inhibition of ATM and/or ATR can significantly increase cancer cell susceptibility to DNA damaging agents, such as ionizing radiation provided by radiolabeled HER3 targeting agents. Accordingly, an object of the invention disclosed herein is to inhibit cancer cells, such as cancer cells that express or overexpress HER3, by administering an inhibitor of ATM (ATMi) and/or an inhibitor of ATR (ATRi) in combination with a HER3 targeting agent. including killing or destroying.

An inhibitor of ATM (ATMi) or an inhibitor of ATR (ATRi) may be an antibody, peptide or small molecule that targets ATM or ATR, respectively. Alternatively, the ATMi or ATRi may reduce or eliminate activation of ATM or ATR by one or more signaling molecules, proteins, or other compounds, or ATM or ATR by all signaling molecules, proteins, or other compounds. reduction or elimination of activation. ATMi and/or ATRi also include compounds that inhibit their expression (eg, compounds that inhibit ATM or ATR transcription or translation). Exemplary ATMi KU-55933 inhibits cell proliferation and induces apoptosis. Other exemplary ATMi include at least KU-59403, wortmannin, CP466722 and KU-60019. Exemplary ATRi include at least Schizandrin B, NU6027, NVP-BEA235, VE-821, VE-822, AZ20 and AZD6738.

exemplary DDRi - Wee1 inhibitor

The checkpoint kinase Wee1 catalyzes the inhibitory phosphorylation of CDK1 (CDC2) and both CDK2 on tyrosine 15, thereby arresting the cell cycle in response to externally induced DNA damage. Deregulated Wee1 expression or activity is considered a hallmark of pathology in several types of cancer. For example, Weel is frequently overexpressed in glioblastoma, malignant melanoma, hepatocellular carcinoma, breast cancer, colon carcinoma, lung carcinoma and head and neck squamous cell carcinoma. Advanced tumors with increased levels of genomic instability may require functional checkpoints capable of repairing such lethal DNA damage. As such, we believe that Weel represents an attractive target in advanced tumors where its inhibition is believed to result in irreversible DNA damage. Accordingly, an object of the invention disclosed by the present application includes administering an inhibitor of Weel in combination with a HER3 targeting agent to inhibit or kill cancer cells, such as cancer cells that express or overexpress HER3.

Wee1 inhibitors can be antibodies, peptides or small molecules that target Wee1. Alternatively, a Weel inhibitor may reduce or eliminate Wee1 activation by one or more signaling molecules, proteins or other compounds, or may result in reduced or eliminated Weel activation by all signaling molecules, proteins or other compounds. there is. The term also includes compounds that reduce or eliminate activation or inactivation of one or more proteins or cell signaling components by Weel (e.g., Weel inhibitors reduce or eliminate Weel-dependent inactivation of cyclins and Cdk activity). can). Wee1 inhibitors also include compounds that inhibit Wee1 expression (eg, compounds that inhibit Wee1 transcription or translation).

Exemplary Weel inhibitors include AZD-1775 (ie, adavosertip), and inhibitors such as, for example, US Pat. No. 7,834,019; 7,935,708; 8,288,396; 8,436,004; 8,710,065; 8,716,297; 8,791,125; 8,796,289; 9,051,327; 9,181,239; 9,714,244; 9,718,821; and 9,850,247; US Publication Nos. US 20100113445 and 20160222459; and International Publication Nos. WO2002090360, 2015019037, 2017013436, 2017216559, 2018011569, and 2018011570.

Additional Wee1 inhibitors include pyrazolopyrimidine derivatives, pyridopyrimidines, 4-(2-chlorophenyl)-9-hydroxypyrrolo[3,4-c]carbazole-1,3-(2H,6H) -Dione (CAS No. 622855-37-2), 6-butyl-4-(2-chlorophenyl)-9-hydroxypyrrolo[3,4-c]carbazole-1,3-(2H,6H )-dione (CAS No. 62285550-9), 4-(2-phenyl)-9-hydroxypyrrolo[3,4-c]carbazole-1,3-(2H,6H)-dione (CAS No. 1177150-89-8), and anti-Weel small interfering RNA (siRNA) molecules.

exemplary DDRi - PARP inhibitor

Another exemplary type of DDRi that may be used is an inhibitor of poly(ADP-ribose) polymerase ("PARP"). Inhibitors of the DNA repair protein PARP, individually and collectively referred to as “PARPi,” have been approved for use in a variety of solid tumors, such as breast and ovarian cancer, particularly in patients with BRCA1/2 mutations. BRCA1 and 2 function in homologous recombination repair (HRR). When mutations occur, they cause genomic instability by shifting the DNA repair process from conservative and precise HRR to DNA end joining that can produce mutations through non-conservative methods such as deletions and insertions.

PARPi was found to exhibit synthetic lethality, as shown by potent single-agent activity in BRCA1/2 mutant cells. This essentially blocks repair of single-stranded DNA breaks. Since HRR does not function in these tumor cells, cell death occurs. Since most tumors do not harbor BRCA1 or BRCA2 mutations, the efficacy of PARPi in such tumors is much less pronounced.

To date, the FDA has approved four PARPi drugs (olaparib, niraparib, rucaparib, and thalazoparib) as monotherapy specifically for patients with germline and somatic mutations in the BRCA1 and BRCA2 genes. Along with veliparib, olaparib, niraparib and rucaparib are among the first-generation PARPi that have entered clinical trials. Their IC50 values were found to be in the nanomolar range. In contrast, second generation PARPi such as thalazoparib have IC50 values in the picomolar range.

All of these PARPi bind to the binding site of b nicotinamide adenine dinucleotide (b-NAD+), a cofactor, in the catalytic domains of PARP1 and PARP2. Enzymes of the PARP family use NAD+ to covalently add poly(ADP-ribose) (PAR) chains onto target proteins, a process called "PARization". PARP1 (the most studied member) and PARP2 are important components of the DNA damage response (DDR) pathway. PARP1 is involved in the repair of single-stranded DNA breaks, and possibly other DNA lesions (Woodhouse, et al.; Krishnakumar, et al.). Through its zinc finger domain, PARP1 binds damaged DNA and then PARates a series of DNA repair effector proteins, releasing nicotinamide as a by-product (Krishnakumar, et al.). Subsequently, PARP1 self-PARylation results in the release of the protein from DNA. However, available PARPi differ in their ability to capture PARP1 on DNA, which appears to correlate with cytotoxicity and drug efficacy. Specifically, drugs such as thalazoparib and olaparib are more effective at capturing PARP1 than veliparib (Murai, et al., 2012; Murai, et al., 2014).

The efficacy of PARPi in ovarian and breast cancer patients with loss-of-function mutations within the BRCA1 or BRCA2 gene is primarily due to the genetic concept of synthetic lethality: the proteins of BRCA 1 and 2 are DNA repair known as homologous recombination repair (HRR). normal maintenance of the integrity of the genome by mediating the process; PARPi normally cause persistent DNA lesions that are otherwise repaired by HR. In the presence of PARPi, PARP1 is captured on DNA delaying the progression of the replication fork. This delay is cytotoxic unless repaired in a timely manner by the HR system. Cells lacking effective HR cannot effectively repair these DNA lesions, so the cells die.

Again, mutations in the BRCA gene and other genes within the HRR system are not prevalent in many cancer types. Thus, to better exploit the therapeutic benefits of PARPi in such cancers, "artificial" synthetic lethality can be induced by pairing PARPi with chemotherapy or radiation therapy. Preclinical studies have demonstrated that combining radiotherapy with PARPi can increase the sensitivity of BRCA1/2 mutant tumor cells to PARP inhibition and expand the sensitivity of non-mutant BRCA tumors to PARP inhibition. Additional studies have shown that ionizing radiation (IR) itself can mediate PARPi synthesis lethality in tumor cells.

Accordingly, an object of the invention disclosed herein is to administer a radiolabeled HER3 targeting agent that delivers ionizing radiation in combination with PARPi.

In various embodiments of the present invention, PARPi can be any known agent that performs that function, preferably one approved by the FDA. Preferably, PARPi is olaparib [Lynparza®], niraparib [Zejula®], rucaparib [Rubraca®] or thalazoparib [Talzenna®] ]am.

Clinically, therapy with PARPi has resulted in sustained antitumor responses in a variety of cancers including ovarian, prostate, pancreatic and triple negative breast cancer (TNBC). In one clinical trial, TNBC patients with germline BRCA1/2 mutations were treated with PARPi, olaparib. Although this regimen clearly demonstrated higher disease stabilization rates in BRCA1/2 mutants compared to non-mutant patients, no sustained response was achieved in either cohort (Gelmon, 2011).

The inventors realized that while this repair pathway is blocked by PARPi, the effect of PARPi can be ameliorated through an increase in dsDNA breakage induced by ionizing radiation provided by HER3 targeting agents. Exemplary PARPi include olaparib, niraparib, rucaparib and thalazoparib.

E. CD47 blockers

Additional agents administered with a HER3 targeting agent may inhibit activity and/or signaling between CD47 (eg, on target cells) and SIRPα (eg, on phagocytic cells) through interaction with a CD47 blocker, such as CD47 or SIRPα. It can be any agent that interferes with or reduces delivery. Non-limiting examples of suitable CD47 blocking agents include CD47 and/or SIRPα reagents including, but not limited to, SIRPα polypeptides, anti-SIRPα antibodies, soluble CD47 polypeptides, and anti-CD47 antibodies or antibody fragments.

Additional examples of CD47 blockers include agents that modulate the expression of CD47 and/or SIRPα. For example, such agents modulate, e.g., block, suppress, reduce, antagonize, neutralize, or otherwise modulate CD47 expression or a nucleic acid approach, such as a phosphorodiamidate morpholino oligomer (PMO) that blocks translation of CD47. antibodies specific for human CD47 that interfere with it. A CD47 antibody or antisense approach may inhibit CD47 expression (eg, inhibit cell surface expression of CD47), inhibit activity and/or signaling, or interfere with the interaction between CD47 and SIRPα. An agent provided herein completely or partially reduces or otherwise modulates CD47 expression or activity when it binds or otherwise interacts with CD47, eg, human CD47. The reduction or modulation of a biological function of CD47 is complete, significant, or partial depending on the interaction between the antibody and human CD47 polypeptides and/or peptides. The agent is capable of reducing the level of CD47 expression or activity in the presence of the antibody by at least 50%, e.g., 60%, when compared to the level of CD47 expression or activity in the absence of interaction, e.g., binding, with an antibody described herein. , 70%, 80%, 90%, 95%, 96%, 98%, 99%, or 100% is considered to inhibit CD47 expression or activity.

According to certain aspects, an anti-CD47 agent specifically binds CD47 (ie, an anti-CD47 antibody) and binds CD47 on one cell (eg, an infected cell) to another cell (eg, a phagocytic cell). ) It is an antibody that reduces the interaction between SIRPα on the. Non-limiting examples of suitable antibodies include clones B6H12, 5F9, 8B6 and C3 and any of those described in International Publication Nos. WO2011/143624 and US Publication No. 20210246206. Suitable anti-CD47 antibodies include fully human, humanized or chimeric versions of such antibodies.

Exemplary human or humanized antibodies that are particularly useful for in vivo application in humans due to their low antigenicity are monoclonal antibodies to at least CD47, such as Hu5F9-G4, a humanized monoclonal available as Magrolimab from Gilead. raw antibodies (Sikic, et al. (2019) Journal of Clinical Oncology 37:946); Remzoparlimab and TJC4 from I-Mab Biopharma; AO-176 from Arch Oncology, Inc.; AK117 from Akesobio Australia Pty; IMC-002 from Innovent Biologics; ZL-1201 from Zia Lab; SHR-1603 from Jiangsu HengRui Medincine Co.; and SRF231 from Surface Oncology. A bispecific monoclonal antibody, such as IBI-322 from Innovent Biologics, which targets both CD47 and PD-L1, is also available. Antibodies to SIRPα such as ALX148 from Alx Oncology; BI 765063 from OSE (OSE-172); as well as small molecule inhibitors such as RRx-001 (1-bromoacetyl-3,3-dinitroazetidine) and azelnidipine (CAS number 123524-52-7) from EpicentRx or their pharmaceutical counterparts. Acceptable salts are also possible. See Table 4 for further description of exemplary agents.

<Table 4>

In addition to inducing tumor phagocytosis by blocking the CD47-SIRPα interaction, AO-176 binds preferentially to tumor cells over normal cells (especially RBCs when binding is negligible) and directly kills tumors compared to normal cells. reported to do

According to certain aspects, a SIRPα reagent may comprise a portion of SIRPα sufficient to bind CD47 with appreciable affinity, normally located between the signal sequence and the transmembrane domain, or a fragment thereof that retains binding activity. A suitable SIRPα reagent reduces (eg, blocks, prevents, etc.) the interaction between the native protein SIRPα and CD47. For example, the CD47 blocker used in various aspects of the present invention is any of those disclosed in U.S. Patent No. 9,969,789, including but not limited to the SIRPα-IgG Fc fusion proteins disclosed therein, such as TTI-621 and TTI -622, both of which preferentially bind CD47 on tumor cells while also binding activating Fc receptors. For example, SEQ ID NO: 116, SEQ ID NO: 117, or a SIRPα-IgG Fc fusion protein comprising the amino acid sequence of SEQ ID NO: 118 can be used.

A therapeutically effective dose of an anti-CD47 antibody or other protein CD47 inhibitor achieves a sustained serum level of the protein of about 40 μg/ml or greater (e.g., about 50 ug/ml or greater, about 60 ug/ml or greater, about 75 ug/ml or greater). ml or more, about 100 ug/ml or more, about 125 ug/ml or more, or about 150 ug/ml or more). A therapeutically effective dose or administration of a CD47 blocker, such as an anti-CD47 antibody or SIRPα fusion protein or small molecule, is, for example, 0.05 - 10 mg/kg (agent weight/subject weight), such as at least 0.1 mg/kg, 0.5 mg/kg kg, 1.0 mg/kg, 1.5 mg/kg, 2.0 mg/kg, 2.5 mg/kg, 3.0 mg/kg, 3.5 mg/kg, 4.0 mg/kg, 4.5 mg/kg, 5.0 mg/kg, 5.5 mg/kg kg, 6.0 mg/kg, 6.5 mg/kg, 7.0 mg/kg, 7.5 mg/kg, 8.0 mg/kg, 8.5 mg/kg, 9.0 mg/kg; or 10 mg/kg, 9.5 mg/kg, 9.0 mg/kg, 8.5 mg/kg, 8.0 mg/kg, 7.5 mg/kg, 7.0 mg/kg, 6.5 mg/kg, 6.0 mg/kg, 5.5 mg/kg , in an amount less than or equal to 5.0 mg/kg, 4.5 mg/kg, 4.0 mg/kg, 3.5 mg/kg, 3.0 mg/kg, 2.5 mg/kg, 2.0 mg/kg, 1.5 mg/kg, 1.0 mg/kg, or Any combination of the upper and lower limits of these values is included. A therapeutically effective dose of a small molecule CD47 blocker as disclosed herein may also be, for example, from 0.01 mg/kg to 1,000 mg/kg and any subrange or value of mg/kg therein, such as from 0.01 mg/kg to 500 mg. /kg or 0.05 mg/kg to 500 mg/kg, or 0.5 mg/kg to 200 mg/kg, or 0.5 mg/kg to 150 mg/kg, or 1.0 mg/kg to 100 mg/kg, or 10 mg/kg kg to 50 mg/kg.

According to certain aspects, an anti-CD47 agent specifically binds SIRPα and reduces the interaction between CD47 on one cell (eg, an infected cell) and SIRPα on another cell (eg, a phagocytic cell). It is a soluble CD47 polypeptide. Suitable soluble CD47 polypeptides can bind SIRPα without activating or stimulating signaling through SIRPα, since activation of SIRPα inhibits phagocytosis. Instead, suitable soluble CD47 polypeptides facilitate preferential phagocytosis of infected cells over non-infected cells. Cells that express higher levels of CD47 compared to normal non-target cells (normal cells) (eg, infected cells) will be preferentially phagocytosed. Thus, a suitable soluble CD47 polypeptide specifically binds to SIRPα without activating/stimulating a signaling response sufficient to inhibit phagocytosis. In some cases, a suitable soluble CD47 polypeptide may be a fusion protein (eg, as described in US Publication No. 20100239579).

Advantageously, CD47 blockers can enhance the cytotoxicity and phagocytosis enhancing effects of radiolabeled targeting agents, such as radiolabeled HER3 and/or HER2 targeting agents, and reduce dose limiting radiotoxicity of the targeting agent, thereby improving tolerability. and allow higher radiation doses of the targeting agent to be used/tolerated in the treatment of the subject.

Example

Example One: radiolabeled HER3 Production of targeting agent

HER3 targeting agents, such as monoclonal antibodies to HER3, are disclosed in International Publication No. WO 2017/155937 and filed on December 9, 2019 entitled "Compositions and Methods for the Preparation of Site-Specific Radioconjugates". It can be labeled with indium-111 ( ¹¹¹ In) or actinium-225 ( ²²⁵ Ac) according to the procedures detailed in US Provisional Application No. 63/042,651.

Radiolabeling : For example, the antibody may be conjugated with a chelate-bearing linker as described, for example, herein or in prior patent applications. Exemplary linkers include at least dodecane tetraacetic acid (DOTA), wherein the goal of the conjugation reaction is to achieve a DOTA-antibody ratio of 3:1 to 5:1. Chelation with the radionuclide ¹¹¹ In or ²²⁵ Ac can then be performed, and the efficiency and purity of the resulting ¹¹¹ In- or ²²⁵ Ac-labeled anti-HER3 antibody can be determined by HPLC and iTLC. .

An exemplary labeling reaction for ²²⁵ Ac is as follows: A reaction containing 15 μl 0.15 M NH ₄ OAc buffer, pH=6.5 and 2 μL (10 μg) DOTA-anti-HER3 (5 mg/ml) was prepared in Eppendorf Mix in a reaction tube and add 4 μL ²²⁵ Ac (10 μCi) in 0.05 M HCl sequentially. The contents of the tube can be mixed with a pipette tip and the reaction mixture can be incubated at 37° C. for 90 minutes while shaking at 100 rpm. At the end of the incubation period, 3 μL of a 1 mM DTPA solution can be added to the reaction mixture and incubated at room temperature for 20 minutes to bind unreacted ²²⁵ Ac to the ²²⁵ Ac-DTPA complex. ²²⁵ Ac-labeled anti-HER3 ( ²²⁵ Ac-DOTA-anti-HER3) was obtained as free ²²⁵ Ac ( ²²⁵ Ac-DTPA ), the radiochemical purity of ²²⁵ Ac-DOTA-anti-HER3 can be determined. In this system, the radiolabeled antibody stays at the point of application and ^{the 225} Ac-DTPA migrates with the solvent front. The strip can be cut in half and counted in a gamma counter equipped with a multi-channel analyzer using channels 72-110 for ²²⁵ Ac to exclude its daughters.

Purification : An exemplary radiolabeled HER3 targeting agent, such as ²²⁵ Ac-DOTA-anti-HER3, was prepared on a PD10 column pre-blocked with 1% HSA or in a 2 x 1.5 mL wash, Viva with a 50 kDa MW cutoff of 3 minutes per spin. It can be purified on a Vivaspin centrifugal concentrator. HPLC analysis of ²²⁵ Ac-DOTA-anti-HER3 after purification was performed using a TSK3000SW XL column eluted with PBS at pH=7.4 and a flow rate of 1 ml/min, Waters equipped with a flow-through Waters UV and Bioscan radiation detector. This can be done using an HPLC system.

Stability Determination : An exemplary radiolabeled HER3 targeting agent, such as ²²⁵ Ac-DOTA-anti-HER3, can be used for stability determination, wherein ²²⁵ Ac-DOTA-anti-HER3 is tested in original volume or in a working buffer (0.15 M NH ₄ OAc), incubated for 48 hours at room temperature (rt) or 96 hours at 4° C. and tested by ITLC. Stability is determined by comparison of intact radiolabeled anti-HER3 before and after incubation. Another antibody labeled with ²²⁵ Ac was found to be stable for up to 96 hours at 4°C.

Immunoreactivity (IR) Determination : Exemplary radiolabeled HER3 targeting agents such as ²²⁵ Ac-DOTA-anti-HER3 can be used in immunoreactivity experiments. HER3 positive cells and control HER3 negative cells can be used in amounts of 1 million to 7.5 million cells per sample to examine the amount of binding (percent radioactive binding to cells after several washes; or immunoreactive fraction (IRF) bead assay). It can be carried out according to the disclosed method as described in the literature [Sharma, 2019] using Previous assays for other antibodies radiolabeled with ¹¹¹ In or ²²⁵ Ac demonstrated about 50-60% immunoreactivity.

Example 2 - Exemplary PARPi Dosage and dosing regimen

(A) olaparib ( Linparza ®) - normal and reduced dosing regimen

Olaparib is marketed by AstraZeneca under the brand name Lynparza®. Lynparza® is sold in tablet form of 100 mg and 150 mg. The dosage is 300 mg taken orally twice daily for a total of 600 mg per day. Administration is continued until disease progression or unacceptable toxicity. This dosing regimen is referred to herein as the “normal” human dosing regimen for Lynparza® regardless of the disorder being treated. Any dosing regimen that has a shorter dosing period (e.g., 21 days) or involves less administration of Lynparza® (e.g., 300 mg/day) is described herein as a "reduced" human dosing regimen. is referred to Examples of reduced human dosing regimens include: (i) 550 mg/day; (ii) 500 mg/day; (iii) 450 mg/day; (iv) 400 mg/day; (v) 350 mg/day; (vi) 300 mg/day; (vii) 250 mg/day; (viii) 200 mg/day; (ix) 150 mg/day; (x) 100 mg/day; or (xi) 50 mg/day.

(B) niraparib ( Zejula ®) - normal and reduced dosing regimen

Niraparib is marketed by Tesaro under the brand name Zejula®. Zejula® is sold in capsule form of 100 mg. The dosage is 300 mg taken orally once daily. Administration is continued until disease progression or unacceptable side effects occur. This dosing regimen is referred to herein as the “normal” human dosing regimen for Zejula® regardless of the disorder being treated. Any dosing regimen that has a shorter dosing period (eg, 21 days) or involves less administration of Zejula® (eg, 150 mg/day) is herein described as a "reduced" human dosing regimen. is referred to Examples of reduced human dosing regimens include: (i) 250 mg/day; (ii) 200 mg/day; (iii) 150 mg/day; (iv) 100 mg/day; or (v) 50 mg/day.

(C) Lucaparib ( Rubraca ®) - normal and reduced dosing regimen

Lucaparib is marketed by Clovis Oncology, Inc. under the trade name Lubraca™. Rubraca™ is sold in tablet form of 200 mg and 300 mg. The dosage is 600 mg taken orally twice daily for a total of 1,200 mg per day. Administration is continued until disease progression or unacceptable toxicity. This dosing regimen is referred to herein as the “normal” human dosing regimen for Rubraca™ regardless of the disorder being treated. Any dosing regimen having a shorter dosing period (e.g., 21 days) or involving less administration of Rubraca™ (e.g., 600 mg/day) is herein referred to as a "reduced" human dosing regimen. is referred to Examples of reduced human dosing regimens include: (i) 1,150 mg/day; (ii) 1,100 mg/day; (iii) 1,050 mg/day; (iv) 1,000 mg/day; (v) 950 mg/day; (vi) 900 mg/day; (vii) 850 mg/day; (viii) 800 mg/day; (ix) 750 mg/day; (x) 700 mg/day; (xi) 650 mg/day; (xii) 600 mg/day; (xiii) 550 mg/day; (xiv) 500 mg/day; (xv) 450 mg/day; (xvi) 400 mg/day; (xvii) 350 mg/day; (xviii) 300 mg/day; (xix) 250 mg/day; (xx) 200 mg/day; (xxi) 150 mg/day; or (xxii) 100 mg/day.

(D)- Thalazoparib ( Talgenna ™) - normal and reduced dosing regimen

Talazoparib is marketed by Pfizer Labs under the trade name Talgenna™. Talgenna™ is sold in 1 mg capsule form. The dosage is 1 mg taken orally. Administration is continued until disease progression or unacceptable toxicity. This dosing regimen is referred to herein as the "normal" human dosing regimen for Talgenna™ regardless of the disorder being treated. Any dosing regimen having a shorter dosing period (eg, 21 days) or involving less administration of Taljenna™ (eg, 0.5 mg/day) is herein described as a "reduced" human dosing regimen. is referred to Examples of reduced human dosing regimens include: (i) 0.9 mg/day; (ii) 0.8 mg/day; (iii) 0.7 mg/day; (iv) 0.6 mg/day; (v) 0.5 mg/day; (vi) 0.4 mg/day; (vii) 0.3 mg/day; (viii) 0.2 mg/day; or (ix) 0.1 mg/day.

Example 3: HER3 targeting agent and to PARPi Dosing regimen for

Human patients can be treated according to the following regimen. One of olaparib, niraparib, rucaparib, or thalazoparib (PARPi) is administered orally according to one of the dosing regimens listed in Example 2, either as single or divided doses, to a radiolabeled HER3 target as described in detail herein. Intravenous administration of topical is involved. For example, a dosing regimen may include, for example: (a) PARPi and the HER3 targeting agent are administered concurrently, wherein (i) each is administered starting on the same day, and (ii) the HER3 targeting agent is administered for at least 1 week. (iii) PARPi is administered daily or twice daily (as appropriate) for a period equal to or greater than the period of administration of the HER3-targeting agent; or (b) the PARPi and the HER3 targeting agent are administered concurrently, wherein (i) the PARPi administration precedes the HER3 targeting agent by at least 1 week, and (ii) the HER3 targeting agent is administered in a single dose or divided by at least 1 week intervals. (iii) PARPi is administered daily or twice daily (if appropriate) for a period of time equal to or greater than the period of administration of the HER3 targeting agent.

Example 4: HER3 Targeting agents and CD47 Dosage regimen for blockers

According to certain aspects of the invention, the CD47 blocking agent can be, for example, a monoclonal antibody that prevents CD47 from binding to SIRPα. Exemplary protein CD47 blockers include magnolimab, remzoparlimab, AO-176, TTI-621, TTI-622, or combinations thereof. A CD47 blocker may alternatively, or additionally, be an agent that modulates the expression of CD47 and/or SIRPα, such as a phosphorodiamidate morpholino oligomer (PMO) that blocks translation of CD47, such as MBT-001 (PMO) , morpholino, sequence: 5'-CGTCACAGGCAGGACCCACTGCCCA-3' [SEQ ID NO: 114]) or the PMO oligomer CD47 disclosed in any of U.S. Pat. No. 8,557,788, U.S. Pat. No. 8,236,313, U.S. Pat. No. 10,370,439 and International Publication No. WO2008060785. any of the inhibitors. A therapeutically effective dose of an anti-CD47 antibody includes at least 0.05 - 10 mg/kg. Thus, the methods of the present invention may comprise administering one or more of an anti-CD47 antibody or other agent concomitantly with intravenous administration of a radiolabeled HER3 targeting agent as described in detail herein in single or divided doses. . For example, a dosing regimen may include, for example: (a) an anti-CD47 antibody or agent and a HER3 targeting agent are administered concurrently, wherein (i) each is administered starting on the same day, and (ii) the HER3 targeting agent is administered concurrently. wherein the agent is administered in single or divided doses at intervals of at least one week, and (iii) the anti-CD47 antibody or agent is administered daily or twice daily (as appropriate) for a period equal to or longer than the period of administration of the HER3 targeting agent. thing; or (b) the anti-CD47 antibody or agent and the HER3 targeting agent are administered concurrently, wherein (i) the anti-CD47 antibody or agent administration precedes the HER3 targeting agent by at least one week, and (ii) the HER3 targeting agent is administered as a single dose or divided dose at intervals of at least 1 week, and (iii) the anti-CD47 antibody or agent is administered daily or twice daily (if appropriate) for a period equal to or longer than the period of administration of the HER3 targeting agent. include

Example 5: HER3 targeting agent and to ICI Dosing regimen for

According to certain aspects of the invention, the immune checkpoint inhibitor (ICI) may be a monoclonal antibody to any of PD-1, PD-L1, PD-L2, CTLA-4, CD137. Therapeutically effective doses of these antibodies include at least 0.05 - 10 mg/kg. Accordingly, the methods of the present invention include administering one or more ICIs accompanied by intravenous administration of a radiolabeled HER3 targeting agent as described in detail herein, in single or divided doses. For example, a dosing regimen may include, for example: (a) the ICI and the HER3-targeting agent are administered concurrently, wherein (i) each is administered starting on the same day, and (ii) the HER3-targeting agent is administered for at least 1 week. (iii) the ICI is administered daily or twice daily (as appropriate) for a period equal to or greater than the period of administration of the HER3-targeting agent; or (b) the ICI and the HER3 targeting agent are administered concurrently, wherein (i) the anti-CD47 antibody administration precedes the HER3 targeting agent by at least 1 week, and (ii) the HER3 targeting agent is administered as a single administration at least 1 week apart. (iii) the ICI is administered daily or twice daily (as appropriate) for a period equal to or greater than the period of administration of the HER3 targeting agent.

Without limitation, the following aspects are also provided by the present disclosure:

Aspect 1. A method of treating solid cancer in a mammalian subject, such as a human patient, comprising administering to the mammalian subject a therapeutically effective amount of a radiolabeled HER3 targeting agent.

Aspect 2. according to any preceding aspect, wherein the solid cancer is breast cancer, gastric cancer, bladder cancer, cervical cancer, endometrial cancer, skin cancer, gastric cancer, testicular cancer, esophageal cancer, bronchioloalveolar cancer, prostate cancer, colorectal cancer, ovarian cancer, cervical cancer skin cancer, pancreatic cancer, lung cancer, kidney cancer, head and neck cancer, or any combination thereof.

Aspect 3. The method of any preceding aspect, wherein the solid cancer is colorectal cancer, gastric cancer, ovarian cancer, non-small cell lung carcinoma, head and neck squamous cell cancer, pancreatic cancer, kidney cancer, or any combination thereof.

Aspect 4. The method of any preceding aspect, wherein the solid cancer is a HER3-positive cancer, such as a HER3-positive solid tumor.

Aspect 5. The method of any preceding aspect, wherein the radiolabeled HER3 targeting agent is ¹³¹ I, ¹²⁵ I, ¹²³ I, ⁹⁰ Y, ¹⁷⁷ Lu, ¹⁸⁶ Re, ¹⁸⁸ Re, ⁸⁹ Sr, ¹⁵³ Sm, ³² P, ²²⁵ Ac, selected from ²¹³ Bi, ²¹³ Po, ²¹¹ At, ²¹² Bi, ²¹³ Bi, ²²³ Ra, ²²⁷ Th, ¹⁴⁹ Tb, ¹³⁷ Cs, ²¹² Pb, ¹⁰³ Pd, or any of those disclosed herein, or any combination thereof A method comprising a radioactive label.

Aspect 6. The method of any preceding aspect, wherein the radiolabeled HER3 targeting agent is selected from ¹³¹ I, ⁹⁰ Y, ¹⁷⁷ Lu, ²²⁵ Ac, ²¹³ Bi, ²¹¹ At, ²¹³ Bi, ²²⁷ Th, ²¹² Pb, or any combination thereof. A method comprising a selected radiolabel.

Aspect 7. The method of any preceding aspect, wherein the radiolabeled HER3 targeting agent comprises an antibody to HER3.

Aspect 8. The method of any preceding aspect, wherein the HER3 targeting agent is an anti-HER3 monoclonal antibody, such as any of those disclosed herein, such as patritumab, serivantumab (MM-121), rumletuzumab, A method comprising a HER3 antibody selected from elgemtumab, GSK2849330 and AV-203, and any combination thereof.

Aspect 9. The HER3 targeting agent of any preceding aspect, wherein the HER3 targeting agent (i) has a heavy chain sequence comprising SEQ ID NO:77 and/or a light chain sequence comprising SEQ ID NO:78; (ii) an immunoglobulin heavy chain variable region comprising CDR-H1 comprising SEQ ID NO: 15, CDR-H2 comprising SEQ ID NO: 16 and/or CDR-H3 comprising SEQ ID NO: 17; and/or an immunoglobulin light chain variable region comprising CDR-L1 comprising SEQ ID NO: 18, CDR-L2 comprising SEQ ID NO: 19, and/or CDR-L3 comprising SEQ ID NO: 20. have; (iii) has an immunoglobulin heavy chain variable region comprising SEQ ID NO: 21 and/or an immunoglobulin light chain variable region comprising SEQ ID NO: 22; or (iv) a monoclonal antibody having the immunoglobulin heavy chain amino acid sequence of SEQ ID NO: 23 and/or the immunoglobulin light chain amino acid sequence of SEQ ID NO: 24.

Aspect 10. The method of any preceding aspect, wherein the HER3 targeting agent comprises a heavy chain variable region having the amino acid sequence set forth in SEQ ID NO:7 and/or a light chain variable region having the amino acid sequence set forth in SEQ ID NO:8 A method comprising a monoclonal antibody comprising

Aspect 11. The method of any preceding aspect, wherein the HER3 targeting agent is selected from the group consisting of one or more of a heavy chain N-terminal region and a complementarity determining region (CDR) having an amino acid sequence as set forth in SEQ ID NOs: 13 and/or 1-3, respectively. and/or comprises a monoclonal antibody comprising one or more of a light chain N-terminal region and CDRs having the amino acid sequence as set forth in SEQ ID NOs: 14 and/or 4-6, respectively.

Aspect 12. The method of any preceding aspect, wherein the effective amount of the radiolabeled HER3 targeting agent is the maximum tolerated dose.

Aspect 13. The method of any preceding aspect, wherein the radiolabeled HER3 targeting agent is ²²⁵ Ac-, ¹⁷⁷ Lu-, or ¹³¹ I-labeled.

Aspect 14. The method of any preceding aspect, wherein the therapeutically effective amount of the radiolabeled HER3 targeting agent comprises a single dose delivering to the subject a dose of less than 2 Gy, or less than 8 Gy, such as from 2 Gy to 8 Gy. .

Aspect 15. The method of any preceding aspect, wherein the radiolabeled HER3 targeting agent is ²²⁵ Ac-labeled, and the effective amount of the ²²⁵ Ac-labeled HER3 targeting agent is from 0.1 to 50 uCi/kg (body weight of the subject), or from 0.2 to 20 uCi/kg (body weight of the subject), or a dose of 0.5 to 10 uCi/kg (body weight of the subject).

Aspect 16. The method of any preceding aspect, wherein the radiolabeled HER3 targeting agent is a ²²⁵ Ac-labeled full-length antibody to HER3, and the effective amount of the ²²⁵ Ac-labeled HER3 targeting agent is 5 uCi/kg (body weight of the subject). ) less than, such as from 0.1 to 5 uCi/kg (body weight of the subject).

Aspect 17. The method of any one of aspects 1 to 6, wherein the radiolabeled HER3 targeting agent is a ²²⁵ Ac-labeled antibody fragment to HER3, such as a minibody or nanobody, and the effective amount of the ²²⁵ Ac-labeled HER3 targeting agent greater than 5 uCi/kg (body weight of the subject), such as between 5 and 20 uCi/kg (body weight of the subject).

Aspect 18. The method of any one of aspects 1 to 14, wherein the radiolabeled HER3 targeting agent is ²²⁵ Ac-labeled, and the effective amount of the ²²⁵ Ac-labeled HER3 targeting agent is between 2 μCi and 2 mCi, or between 2 μCi and 250 μCi. , or from 75 μCi to 400 μCi.

Aspect 19. The method of any one of aspects 1 to 14, wherein the radiolabeled HER3 targeting agent is ¹⁷⁷ Lu-labeled and the effective amount of the HER3 targeting agent is a dose of less than 1000 uCi/kg (body weight of the subject), such as 1 to 900 uCi/kg (body weight of the subject), or 5 to 250 uCi/kg (body weight of the subject) or 50 to 450 uCi/kg (body weight).

Aspect 20. The method of any one of aspects 1 to 14, wherein the radiolabeled HER3 targeting agent is ¹⁷⁷ Lu-labeled, and the effective amount of the ¹⁷⁷ Lu-labeled HER3 targeting agent is between 10 mCi and 30 mCi or less, or at least 100 A method comprising a dose of μCi to 3 mCi or less, or 3 mCi to 30 mCi or less.

Aspect 21. The method of any one of aspects 1 to 14, wherein the radiolabeled HER3 targeting agent is ¹³¹ I-labeled, and the effective amount of the ¹³¹ I-labeled HER3 targeting agent is at a dose less than 1200 mCi, such as from 25 to 1200 mCi; or a dose of 100 to 400 mCi, or 300 to 600 mCi, or 500 to 1000 mCi.

Aspect 22. The method of any one of aspects 1 to 14, wherein the radiolabeled HER3 targeting agent is ¹³¹ I-labeled, and the effective amount of the ¹³¹ I-labeled HER3 targeting agent is at a dose of less than 200 mCi, such as 1 to 200 mCi; Or a method comprising a dose of 25 to 175 mCi, or 50 to 150 mCi.

Aspect 23. The method of any preceding aspect, wherein the effective amount of the HER3 targeting agent is less than 3 mg/kg (subject's body weight), such as 0.001 mg/kg (patient's body weight) to 3.0 mg/kg (patient's body weight), or 0.005 mg/kg (patient's body weight). kg (patient body weight) to 2.0 mg/kg (patient body weight), or 0.01 mg/kg (patient body weight) to 1 mg/kg (patient body weight), or 0.1 mg/kg (patient body weight) to 0.6 mg/kg (patient body weight) body weight), or 0.3 mg/kg (patient body weight), or 0.4 mg/kg (patient body weight), or 0.5 mg/kg (patient body weight), or 0.6 mg/kg (patient body weight) method.

Aspect 24. The group of any preceding aspect, wherein the HER3 targeting agent consists of once every 7 days, 10 days, 12 days, 14 days, 20 days, 24 days, 28 days, 36 days and 42 days throughout the treatment period wherein the treatment period comprises at least two administrations.

Aspect 25. The method of any one of aspects 1-6, wherein the HER3 targeting agent is a peptide or small molecule.

Aspect 26. The method of any preceding aspect, further comprising administering to the subject a therapeutically effective amount of an immune checkpoint therapy, a chemotherapeutic agent, a DNA damage response inhibitor (DDRi), a CD47 blocker, or a combination thereof.

Aspect 27. The method of aspect 26, wherein the immune checkpoint therapy is CTLA-4, PD-1, TIM-3, VISTA, BTLA, LAG-3, TIGIT, CD28, OX40, GITR, CD137, CD40, CD40L, CD27, HVEM , PD-L1, PD-L2, PD-L3, PD-L4, CD80, CD86, CD137-L, GITR-L, CD226, B7-H3, B7-H4, BTLA, TIGIT, GALS, KIR, 2B4, CD160 , or an antibody to CGEN-15049 or other blocking agent, or any combination of such antibody and blocking agent.

Aspect 28. The method of aspect 27, wherein the immune checkpoint therapy comprises an antibody to PD-1, PD-L1, PD-L2, CTLA-4, CD137, or a combination thereof.

Aspect 29. The method of aspect 26, wherein the DDRi is a poly(ADP-ribose) polymerase inhibitor (PARPi), a telangiectatic ataxia mutated inhibitor (ATMi), a telangiectatic ataxia mutated and a Rad-3 related inhibitor (ATRi), or a Wee1 inhibitor.

Aspect 30. The method of aspect 29, wherein the PARPi comprises one or more of olaparib, niraparib, rucaparib and thalazoparib.

Aspect 31. The method of aspect 29, wherein the ATMi comprises one or more of KU-55933, KU-59403, wortmannin, CP466722, or KU-60019.

Aspect 32. The method of aspect 29, wherein the ATRi comprises one or more of Schizandrin B, NU6027, NVP-BEA235, VE-821, VE-822, AZ20, or AZD6738.

Aspect 33. The method of aspect 29, wherein the Weel inhibitor comprises AZD-1775 (ie, Adavosertip).

Aspect 34. The method of aspect 26, wherein the CD47 blocker comprises an agent that prevents CD47 from binding to SIRPα, such as a monoclonal antibody and/or an agent that modulates CD47 expression.

Aspect 35. The method of aspect 34, wherein the CD47 blocker comprises one or more of magrolimab, remzoparlimab, AO-176, TTI-621, TTI-622, or combinations thereof; Wherein the agent that modulates CD47 expression comprises a phosphorodiamidate morpholino oligomer (PMO) that reduces expression of CD47 (eg, MBT-001).

Aspect 36. The method of aspect 34, wherein the therapeutically effective amount of the CD47 blocker comprises 0.05 to 5 mg/kg of patient body weight.

Aspect 37. The method of aspect 26, wherein the HER3 targeting agent is administered at least 1 week prior to administration of the immune checkpoint therapy and/or DDRi and/or CD47 blocker; or the immune checkpoint therapy and/or DDRi and/or CD47 blocker is administered at least one week prior to administration of the HER3 targeting agent.

Aspect 38. The method of aspect 26, wherein the HER3 targeting agent is administered with one of the immune checkpoint therapy or DDRi or CD47 blocker, and the other of the immune checkpoint therapy or DDRi or CD47 blocker is administered before or after administration of the HER3 targeting agent. how it would be.

Aspect 39. The method of aspect 26, wherein the HER3 targeting agent is administered concurrently with the immune checkpoint therapy and/or DDRi and/or CD47 blocker.

Aspect 40. The method of any preceding aspect, wherein the HER3 targeting agent is a multispecific antibody, wherein the multispecific antibody comprises a first target recognition component that specifically binds an epitope of HER3, and a HER3 target recognition component different from the first target recognition component. A method comprising a second target recognition component that specifically binds to an epitope of, or an epitope of a different antigen.

Aspect 41. The method of aspect 40, wherein the HER3 targeting agent is a bispecific antibody to HER3/HER2, such as MM-111 or MCLA0-128, or a bispecific antibody to IGF-1R/HER3, such as MM-141 (ie , istiratumab), and/or a bispecific antibody to HER1/HER3, such as MEHD7945A (ie, duligotumab).

Aspect 42. diagnosing the subject as having HER3-positive cells; and if the subject has HER3-positive cells, a method of treating a proliferative disease or disorder comprising administering to the subject a therapeutically effective amount of a HER3 targeting agent according to any of the methods of aspects 1-41.

Aspect 43. The method of aspect 42, wherein the diagnosing step comprises obtaining a blood or tissue sample from the subject; immobilizing the sample on a substrate; and detecting the presence or absence of the HER3 antigen using a diagnostic antibody, wherein the diagnostic antibody is a radiolabel, such as ³ H, ¹⁴ C, ³² P, ³⁵ S, and ¹²⁵ I; fluorescent or chemiluminescent compounds such as fluorescein isothiocyanate, rhodamine or luciferin; or an antibody to HER3 labeled with an enzyme such as alkaline phosphatase, β-galactosidase or horseradish peroxidase.

Aspect 44. The method of aspect 42, wherein the diagnosing step is administering a HER3 targeting agent to the subject, wherein the HER3 targeting agent is ¹⁸ F, ¹¹ C, ⁶⁸ Ga, ⁶⁴ Cu, ⁸⁹ Zr, ¹²⁴ I, ^99m Tc, or ¹¹¹ In To contain a radiolabel selected from the group containing; waiting sufficient time for the HER3 targeting agent to accumulate at the tissue site; and imaging the tissue with a non-invasive imaging technique to detect the presence or absence of HER3-positive cells.

Aspect 45. The method of aspect 44, wherein the non-invasive imaging technique comprises positron emission tomography (PET imaging) for ¹⁸ F, ¹¹ C, ⁶⁸ Ga, ⁶⁴ Cu, ⁸⁹ Zr, or ¹²⁴ I labeled HER3 targeting agents. or, in the case of ^99m Tc or ¹¹¹ In labeled HER3 targeting agents, single photon emission computed tomography (SPECT imaging).

While various specific embodiments have been illustrated and described herein, it will be appreciated that various changes may be made without departing from the spirit and scope of the invention(s). Moreover, features described in connection with one aspect of the invention may be used in connection with other aspects of the invention even if not explicitly illustrated in combination herein.

references

Mishra R, Patel H, Alanazi S, Yuan L, Garrett JT. HER3 signaling and targeted therapy in cancer. Oncol Rev. 2018;12(1).

Meneses-Lorente G, Friess T, Kolm I, et al. Preclinical pharmacokinetics, pharmacodynamics, and efficacy of RG7116: a novel humanized, glycoengineered anti-HER3 antibody. Cancer Chemother Pharmacol. 2015;75(4):837-850.

Mirschberger C, Schiller CB, Schraml M, et al. RG7116, a Therapeutic Antibody That Binds the Inactive HER3 Receptor and Is Optimized for Immune Effector Activation. Cancer Res. 2013;73(16):5183-5194.

Meulendijks D, Jacob W, Martinez-Garcia M, et al. First-in-Human Phase I Study of Lumretuzumab, a Glycoengineered Humanized Anti-HER3 Monoclonal Antibody, in Patients with Metastatic or Advanced HER3-Positive Solid Tumors. Clin Cancer Res. 2016;22(4):877-885.

Reynolds KL, Bedard PL, Lee S-H, et al. A phase I open-label dose-escalation study of the anti-HER3 monoclonal antibody LJM716 in patients with advanced squamous cell carcinoma of the esophagus or head and neck and HER2-overexpressing breast or gastric cancer. BMC Cancer. 2017;17(1):646.

SEQUENCE LISTING <110> ACTINIUM PHARMACEUTICALS, INC. <120> HER3 RADIOIMMUNOTHERAPY FOR THE TREATMENT OF SOLID CANCERS <130> ATNM-010PCT <140> <141> <150> PCT/US21/56259 <151> 2021-10-22 <150> US 63/250,725 <151> 2021-09-30 <150> US 63/226,699 <151> 2021-07-28 <150> US 63/118,181 <151> 2020-11-25 <150> US 63/116,225 <151> 2020-11-20 <160> 118 <170> PatentIn version 3.5 <210> 1 <211> 5 <212> PRT 213 <213> <400> 1 Ser His Trp Leu His 1 5 <210> 2 <211> 17 <212> PRT 213 <213> <400> 2 Val Leu Asp Pro Ser Asp Phe Tyr Ser Asn Tyr Asn Gln Asn Phe Lys 1 5 10 15 Gly <210> 3 <211> 11 <212> PRT 213 <213> <400> 3 Gly Leu Leu Ser Gly Asp Tyr Ala Met Asp Tyr 1 5 10 <210> 4 <211> 16 <212> PRT 213 <213> <400> 4 Arg Ser Ser Gln Ser Ile Val His Ser Asn Gly Asn Thr Tyr Leu Glu 1 5 10 15 <210> 5 <211> 7 <212> PRT 213 <213> <400> 5 Lys Val Ser Asn Arg Phe Ser 1 5 <210> 6 <211> 9 <212> PRT 213 <213> <400> 6 Phe Gln Gly Ser Tyr Val Pro Trp Thr 1 5 <210> 7 <211> 120 <212> PRT 213 <213> <400> 7 Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Arg Pro Gly Thr 1 5 10 15 Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser His 20 25 30 Trp Leu His Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Val Leu Asp Pro Ser Asp Phe Tyr Ser Asn Tyr Asn Gln Asn Phe 50 55 60 Lys Gly Lys Ala Thr Leu Thr Val Asp Thr Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Leu Leu Ser Gly Asp Tyr Ala Met Asp Tyr Trp Gly Ala 100 105 110 Gly Thr Ser Val Thr Val Ser Ser 115 120 <210> 8 <211> 112 <212> PRT 213 <213> <400> 8 Asp Val Leu Met Thr Gln Ile Pro Leu Ser Leu Pro Val Ser Leu Gly 1 5 10 15 Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser 20 25 30 Asn Gly Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Lys Ser Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys Phe Gln Gly 85 90 95 Ser Tyr Val Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 <210> 9 <211> 463 <212> PRT 213 <213> <400> 9 Met Gly Trp Ser Cys Ile Ile Val Leu Leu Val Ser Thr Ala Thr Gly 1 5 10 15 Val His Ser Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Arg 20 25 30 Pro Gly Thr Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe 35 40 45 Thr Ser His Trp Leu His Trp Val Lys Gln Arg Pro Gly Gln Gly Leu 50 55 60 Glu Trp Ile Gly Val Leu Asp Pro Ser Asp Phe Tyr Ser Asn Tyr Asn 65 70 75 80 Gln Asn Phe Lys Gly Lys Ala Thr Leu Thr Val Asp Thr Ser Ser Ser 85 90 95 Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val 100 105 110 Tyr Tyr Cys Ala Arg Gly Leu Leu Ser Gly Asp Tyr Ala Met Asp Tyr 115 120 125 Trp Gly Gln Gly Thr Ser Val Thr Val Ser Ser Ala Lys Thr Thr Pro 130 135 140 Pro Ser Val Tyr Pro Leu Ala Pro Gly Ser Ala Ala Gln Thr Asn Ser 145 150 155 160 Met Val Thr Leu Gly Cys Leu Val Lys Gly Tyr Phe Pro Glu Pro Val 165 170 175 Thr Val Thr Trp Asn Ser Gly Ser Leu Ser Ser Gly Val His Thr Phe 180 185 190 Pro Ala Val Leu Gln Ser Asp Leu Tyr Thr Leu Ser Ser Ser Val Thr 195 200 205 Val Pro Ser Ser Thr Trp Pro Ser Gln Thr Val Thr Cys Asn Val Ala 210 215 220 His Pro Ala Ser Ser Thr Lys Val Asp Lys Lys Ile Val Pro Arg Asp 225 230 235 240 Cys Gly Cys Lys Pro Cys Ile Cys Thr Val Pro Glu Val Ser Ser Val 245 250 255 Phe Ile Phe Pro Pro Lys Pro Lys Asp Val Leu Thr Ile Thr Leu Thr 260 265 270 Pro Lys Val Thr Cys Val Val Val Asp Ile Ser Lys Asp Asp Pro Glu 275 280 285 Val Gln Phe Ser Trp Phe Val Asp Asp Val Glu Val His Thr Ala Gln 290 295 300 Thr Gln Pro Arg Glu Glu Gln Phe Asn Ser Thr Phe Arg Ser Val Ser 305 310 315 320 Glu Leu Pro Ile Met His Gln Asp Trp Leu Asn Gly Lys Glu Phe Lys 325 330 335 Cys Arg Val Asn Ser Ala Ala Phe Pro Ala Pro Ile Glu Lys Thr Ile 340 345 350 Ser Lys Thr Lys Gly Arg Pro Lys Ala Pro Gln Val Tyr Thr Ile Pro 355 360 365 Pro Pro Lys Glu Gln Met Ala Lys Asp Lys Val Ser Leu Thr Cys Met 370 375 380 Ile Thr Asp Phe Phe Pro Glu Asp Ile Thr Val Glu Trp Gln Trp Asn 385 390 395 400 Gly Gln Pro Ala Glu Asn Tyr Lys Asn Thr Gln Pro Ile Met Asp Thr 405 410 415 Asp Gly Ser Tyr Phe Val Tyr Ser Lys Leu Asn Val Gln Lys Ser Asn 420 425 430 Trp Glu Ala Gly Asn Thr Phe Thr Cys Ser Val Leu His Glu Gly Leu 435 440 445 His Asn His His Thr Glu Lys Ser Leu Ser His Ser Pro Gly Lys 450 455 460 <210> 10 <211> 238 <212> PRT 213 <213> <400> 10 Met Lys Leu Pro Val Arg Leu Leu Val Leu Met Phe Trp Ile Pro Ala 1 5 10 15 Ser Ser Ser Asp Val Leu Met Thr Gln Ile Pro Leu Ser Leu Pro Val 20 25 30 Ser Leu Gly Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile 35 40 45 Val His Ser Asn Gly Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro 50 55 60 Gly Gln Ser Pro Lys Ser Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser 65 70 75 80 Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr 85 90 95 Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys 100 105 110 Phe Gln Gly Ser Tyr Val Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu 115 120 125 Glu Ile Lys Arg Ala Asp Ala Ala Pro Thr Val Ser Ile Phe Pro Pro 130 135 140 Ser Ser Glu Gln Leu Thr Ser Gly Gly Ala Ser Val Val Cys Phe Leu 145 150 155 160 Asn Asn Phe Tyr Pro Arg Asp Ile Asn Val Lys Trp Lys Ile Asp Gly 165 170 175 Ser Glu Arg Gln Asn Gly Val Leu Asn Ser Trp Thr Asp Gln Asp Ser 180 185 190 Lys Asp Ser Thr Tyr Ser Met Ser Ser Thr Leu Thr Leu Thr Lys Asp 195 200 205 Glu Tyr Glu Arg His Asn Ser Tyr Thr Cys Glu Ala Thr His Lys Thr 210 215 220 Ser Thr Ser Pro Ile Val Lys Ser Phe Asn Arg Asn Glu Cys 225 230 235 <210> 11 <211> 444 <212> PRT 213 <213> <400> 11 Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Arg Pro Gly Thr 1 5 10 15 Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser His 20 25 30 Trp Leu His Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Val Leu Asp Pro Ser Asp Phe Tyr Ser Asn Tyr Asn Gln Asn Phe 50 55 60 Lys Gly Lys Ala Thr Leu Thr Val Asp Thr Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Leu Leu Ser Gly Asp Tyr Ala Met Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Ser Val Thr Val Ser Ser Ala Lys Thr Thr Pro Pro Ser Val 115 120 125 Tyr Pro Leu Ala Pro Gly Ser Ala Ala Gln Thr Asn Ser Met Val Thr 130 135 140 Leu Gly Cys Leu Val Lys Gly Tyr Phe Pro Glu Pro Val Thr Val Thr 145 150 155 160 Trp Asn Ser Gly Ser Leu Ser Ser Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu Gln Ser Asp Leu Tyr Thr Leu Ser Ser Ser Val Thr Val Pro Ser 180 185 190 Ser Thr Trp Pro Ser Gln Thr Val Thr Cys Asn Val Ala His Pro Ala 195 200 205 Ser Ser Thr Lys Val Asp Lys Lys Ile Val Pro Arg Asp Cys Gly Cys 210 215 220 Lys Pro Cys Ile Cys Thr Val Pro Glu Val Ser Ser Val Phe Ile Phe 225 230 235 240 Pro Pro Lys Pro Lys Asp Val Leu Thr Ile Thr Leu Thr Pro Lys Val 245 250 255 Thr Cys Val Val Val Asp Ile Ser Lys Asp Asp Pro Glu Val Gln Phe 260 265 270 Ser Trp Phe Val Asp Asp Val Glu Val His Thr Ala Gln Thr Gln Pro 275 280 285 Arg Glu Glu Gln Phe Asn Ser Thr Phe Arg Ser Val Ser Glu Leu Pro 290 295 300 Ile Met His Gln Asp Trp Leu Asn Gly Lys Glu Phe Lys Cys Arg Val 305 310 315 320 Asn Ser Ala Ala Phe Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr 325 330 335 Lys Gly Arg Pro Lys Ala Pro Gln Val Tyr Thr Ile Pro Pro Pro Lys 340 345 350 Glu Gln Met Ala Lys Asp Lys Val Ser Leu Thr Cys Met Ile Thr Asp 355 360 365 Phe Phe Pro Glu Asp Ile Thr Val Glu Trp Gln Trp Asn Gly Gln Pro 370 375 380 Ala Glu Asn Tyr Lys Asn Thr Gln Pro Ile Met Asp Thr Asp Gly Ser 385 390 395 400 Tyr Phe Val Tyr Ser Lys Leu Asn Val Gln Lys Ser Asn Trp Glu Ala 405 410 415 Gly Asn Thr Phe Thr Cys Ser Val Leu His Glu Gly Leu His Asn His 420 425 430 His Thr Glu Lys Ser Leu Ser His Ser Pro Gly Lys 435 440 <210> 12 <211> 219 <212> PRT 213 <213> <400> 12 Asp Val Leu Met Thr Gln Ile Pro Leu Ser Leu Pro Val Ser Leu Gly 1 5 10 15 Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser 20 25 30 Asn Gly Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Lys Ser Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys Phe Gln Gly 85 90 95 Ser Tyr Val Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 Arg Ala Asp Ala Ala Pro Thr Val Ser Ile Phe Pro Pro Ser Ser Glu 115 120 125 Gln Leu Thr Ser Gly Gly Ala Ser Val Val Cys Phe Leu Asn Asn Phe 130 135 140 Tyr Pro Arg Asp Ile Asn Val Lys Trp Lys Ile Asp Gly Ser Glu Arg 145 150 155 160 Gln Asn Gly Val Leu Asn Ser Trp Thr Asp Gln Asp Ser Lys Asp Ser 165 170 175 Thr Tyr Ser Met Ser Ser Thr Leu Thr Leu Thr Lys Asp Glu Tyr Glu 180 185 190 Arg His Asn Ser Tyr Thr Cys Glu Ala Thr His Lys Thr Ser Thr Ser 195 200 205 Pro Ile Val Lys Ser Phe Asn Arg Asn Glu Cys 210 215 <210> 13 <211> 10 <212> PRT 213 <213> <400> 13 Gln Val Gln Leu Gln Gln Pro Gly Ala Glu 1 5 10 <210> 14 <211> 10 <212> PRT 213 <213> <400> 14 Asp Val Leu Met Thr Gln Ile Pro Leu Ser 1 5 10 <210> 15 <211> 5 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 15 Asp Tyr Ala Met Ser 1 5 <210> 16 <211> 17 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 16 Thr Ile Ser Asp Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val Lys 1 5 10 15 Gly <210> 17 <211> 10 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 17 Glu Trp Gly Asp Tyr Asp Gly Phe Asp Tyr 1 5 10 <210> 18 <211> 11 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 18 Arg Ala Ser Gln Glu Ile Ser Gly Tyr Leu Ser 1 5 10 <210> 19 <211> 7 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 19 Ala Ala Ser Thr Leu Asp Ser 1 5 <210> 20 <211> 9 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 20 Leu Gln Tyr Asp Ser Tyr Pro Tyr Thr 1 5 <210> 21 <211> 119 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 21 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Ala Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Thr Ile Ser Asp Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Trp Gly Asp Tyr Asp Gly Phe Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 22 <211> 107 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 22 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Glu Ile Ser Gly Tyr 20 25 30 Leu Ser Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Arg Leu Ile 35 40 45 Tyr Ala Ala Ser Thr Leu Asp Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln Tyr Asp Ser Tyr Pro Tyr 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 23 <211> 471 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 23 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Gln Val Gln Leu Val Glu Ser Gly Gly Gly 20 25 30 Leu Val Lys Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly 35 40 45 Phe Thr Phe Ser Asp Tyr Ala Met Ser Trp Ile Arg Gln Ala Pro Gly 50 55 60 Lys Gly Leu Glu Trp Val Ser Thr Ile Ser Asp Gly Gly Thr Tyr Thr 65 70 75 80 Tyr Tyr Pro Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn 85 90 95 Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp 100 105 110 Thr Ala Val Tyr Tyr Cys Ala Arg Glu Trp Gly Asp Tyr Asp Gly Phe 115 120 125 Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr 130 135 140 Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser 145 150 155 160 Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu 165 170 175 Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His 180 185 190 Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser 195 200 205 Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys 210 215 220 Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu 225 230 235 240 Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 245 250 255 Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 260 265 270 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 275 280 285 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 290 295 300 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 305 310 315 320 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 325 330 335 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 340 345 350 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 355 360 365 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys 370 375 380 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 385 390 395 400 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 405 410 415 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 420 425 430 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 435 440 445 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 450 455 460 Leu Ser Leu Ser Pro Gly Lys 465 470 <210> 24 <211> 236 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 24 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Asp Ile Gln Met Thr Gln Ser Pro Ser Ser 20 25 30 Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser 35 40 45 Gln Glu Ile Ser Gly Tyr Leu Ser Trp Tyr Gln Gln Lys Pro Gly Lys 50 55 60 Ala Pro Lys Arg Leu Ile Tyr Ala Ala Ser Thr Leu Asp Ser Gly Val 65 70 75 80 Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr 85 90 95 Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln 100 105 110 Tyr Asp Ser Tyr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile 115 120 125 Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp 130 135 140 Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn 145 150 155 160 Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu 165 170 175 Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp 180 185 190 Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr 195 200 205 Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser 210 215 220 Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 225 230 235 <210> 25 <211> 5 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 25 Ser His Trp Leu His 1 5 <210> 26 <211> 17 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 26 Val Leu Asp Pro Ser Asp Phe Tyr Ser Asn Tyr Asn Gln Asn Phe Lys 1 5 10 15 Gly <210> 27 <211> 11 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 27 Gly Leu Leu Ser Gly Asp Tyr Ala Met Asp Tyr 1 5 10 <210> 28 <211> 16 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 28 Arg Ser Ser Gln Ser Ile Val His Ser Asn Gly Asn Thr Tyr Leu Glu 1 5 10 15 <210> 29 <211> 7 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 29 Lys Val Ser Asn Arg Phe Ser 1 5 <210> 30 <211> 9 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 30 Phe Gln Gly Ser Tyr Val Pro Trp Thr 1 5 <210> 31 <211> 120 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 31 Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Arg Pro Gly Thr 1 5 10 15 Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser His 20 25 30 Trp Leu His Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Val Leu Asp Pro Ser Asp Phe Tyr Ser Asn Tyr Asn Gln Asn Phe 50 55 60 Lys Gly Lys Ala Thr Leu Thr Val Asp Thr Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Leu Leu Ser Gly Asp Tyr Ala Met Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Ser Val Thr Val Ser Ser 115 120 <210> 32 <211> 112 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 32 Asp Val Leu Met Thr Gln Ile Pro Leu Ser Leu Pro Val Ser Leu Gly 1 5 10 15 Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser 20 25 30 Asn Gly Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Lys Ser Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys Phe Gln Gly 85 90 95 Ser Tyr Val Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 <210> 33 <211> 463 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 33 Met Gly Trp Ser Cys Ile Ile Val Leu Leu Val Ser Thr Ala Thr Gly 1 5 10 15 Val His Ser Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Arg 20 25 30 Pro Gly Thr Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe 35 40 45 Thr Ser His Trp Leu His Trp Val Lys Gln Arg Pro Gly Gln Gly Leu 50 55 60 Glu Trp Ile Gly Val Leu Asp Pro Ser Asp Phe Tyr Ser Asn Tyr Asn 65 70 75 80 Gln Asn Phe Lys Gly Lys Ala Thr Leu Thr Val Asp Thr Ser Ser Ser 85 90 95 Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val 100 105 110 Tyr Tyr Cys Ala Arg Gly Leu Leu Ser Gly Asp Tyr Ala Met Asp Tyr 115 120 125 Trp Gly Gln Gly Thr Ser Val Thr Val Ser Ser Ala Lys Thr Thr Pro 130 135 140 Pro Ser Val Tyr Pro Leu Ala Pro Gly Ser Ala Ala Gln Thr Asn Ser 145 150 155 160 Met Val Thr Leu Gly Cys Leu Val Lys Gly Tyr Phe Pro Glu Pro Val 165 170 175 Thr Val Thr Trp Asn Ser Gly Ser Leu Ser Ser Gly Val His Thr Phe 180 185 190 Pro Ala Val Leu Gln Ser Asp Leu Tyr Thr Leu Ser Ser Ser Val Thr 195 200 205 Val Pro Ser Ser Thr Trp Pro Ser Gln Thr Val Thr Cys Asn Val Ala 210 215 220 His Pro Ala Ser Ser Thr Lys Val Asp Lys Lys Ile Val Pro Arg Asp 225 230 235 240 Cys Gly Cys Lys Pro Cys Ile Cys Thr Val Pro Glu Val Ser Ser Val 245 250 255 Phe Ile Phe Pro Pro Lys Pro Lys Asp Val Leu Thr Ile Thr Leu Thr 260 265 270 Pro Lys Val Thr Cys Val Val Val Asp Ile Ser Lys Asp Asp Pro Glu 275 280 285 Val Gln Phe Ser Trp Phe Val Asp Asp Val Glu Val His Thr Ala Gln 290 295 300 Thr Gln Pro Arg Glu Glu Gln Phe Asn Ser Thr Phe Arg Ser Val Ser 305 310 315 320 Glu Leu Pro Ile Met His Gln Asp Trp Leu Asn Gly Lys Glu Phe Lys 325 330 335 Cys Arg Val Asn Ser Ala Ala Phe Pro Ala Pro Ile Glu Lys Thr Ile 340 345 350 Ser Lys Thr Lys Gly Arg Pro Lys Ala Pro Gln Val Tyr Thr Ile Pro 355 360 365 Pro Pro Lys Glu Gln Met Ala Lys Asp Lys Val Ser Leu Thr Cys Met 370 375 380 Ile Thr Asp Phe Phe Pro Glu Asp Ile Thr Val Glu Trp Gln Trp Asn 385 390 395 400 Gly Gln Pro Ala Glu Asn Tyr Lys Asn Thr Gln Pro Ile Met Asp Thr 405 410 415 Asp Gly Ser Tyr Phe Val Tyr Ser Lys Leu Asn Val Gln Lys Ser Asn 420 425 430 Trp Glu Ala Gly Asn Thr Phe Thr Cys Ser Val Leu His Glu Gly Leu 435 440 445 His Asn His His Thr Glu Lys Ser Leu Ser His Ser Pro Gly Lys 450 455 460 <210> 34 <211> 238 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 34 Met Lys Leu Pro Val Arg Leu Leu Val Leu Met Phe Trp Ile Pro Ala 1 5 10 15 Ser Ser Ser Asp Val Leu Met Thr Gln Ile Pro Leu Ser Leu Pro Val 20 25 30 Ser Leu Gly Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile 35 40 45 Val His Ser Asn Gly Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro 50 55 60 Gly Gln Ser Pro Lys Ser Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser 65 70 75 80 Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr 85 90 95 Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys 100 105 110 Phe Gln Gly Ser Tyr Val Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu 115 120 125 Glu Ile Lys Arg Ala Asp Ala Ala Pro Thr Val Ser Ile Phe Pro Pro 130 135 140 Ser Ser Glu Gln Leu Thr Ser Gly Gly Ala Ser Val Val Cys Phe Leu 145 150 155 160 Asn Asn Phe Tyr Pro Arg Asp Ile Asn Val Lys Trp Lys Ile Asp Gly 165 170 175 Ser Glu Arg Gln Asn Gly Val Leu Asn Ser Trp Thr Asp Gln Asp Ser 180 185 190 Lys Asp Ser Thr Tyr Ser Met Ser Ser Thr Leu Thr Leu Thr Lys Asp 195 200 205 Glu Tyr Glu Arg His Asn Ser Tyr Thr Cys Glu Ala Thr His Lys Thr 210 215 220 Ser Thr Ser Pro Ile Val Lys Ser Phe Asn Arg Asn Glu Cys 225 230 235 <210> 35 <211> 7 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 35 Thr Phe Gly Leu Ser Val Gly 1 5 <210> 36 <211> 16 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 36 His Ile Trp Trp Asp Asp Asp Lys Tyr Tyr Asn Pro Ala Leu Lys Ser 1 5 10 15 <210> 37 <211> 10 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 37 Ile Gly Ala Asp Ala Leu Pro Phe Asp Tyr 1 5 10 <210> 38 <211> 16 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 38 Arg Ser Ser Lys Ser Leu Leu His Ser Asn Gly Asn Thr Tyr Leu Tyr 1 5 10 15 <210> 39 <211> 7 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 39 Arg Met Ser Asn Leu Ala Ser 1 5 <210> 40 <211> 9 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 40 Met Gln His Leu Glu Tyr Pro Phe Thr 1 5 <210> 41 <211> 120 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 41 Gln Val Thr Leu Lys Glu Ser Gly Pro Gly Ile Leu Arg Pro Ser Gln 1 5 10 15 Thr Leu Ser Leu Thr Cys Ser Phe Ser Gly Phe Ser Leu Ser Thr Phe 20 25 30 Gly Leu Ser Val Gly Trp Ile Arg Gln Pro Ser Gly Lys Gly Leu Glu 35 40 45 Trp Leu Ala His Ile Trp Trp Asp Asp Asp Lys Tyr Tyr Asn Pro Ala 50 55 60 Leu Lys Ser Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys Asn Gln Val 65 70 75 80 Phe Leu Lys Ile Ala Asn Val Asp Thr Ala Asp Thr Ala Thr Tyr Tyr 85 90 95 Cys Ala Arg Ile Gly Ala Asp Ala Leu Pro Phe Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Thr Leu Thr Val Ser Ser 115 120 <210> 42 <211> 112 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 42 Asp Ile Val Leu Thr Gln Thr Ala Pro Ser Val Pro Val Thr Pro Gly 1 5 10 15 Glu Ser Val Ser Ile Ser Cys Arg Ser Ser Lys Ser Leu Leu His Ser 20 25 30 Asn Gly Asn Thr Tyr Leu Tyr Trp Phe Leu Gln Arg Pro Gly Gln Ser 35 40 45 Pro Gln Leu Leu Ile Tyr Arg Met Ser Asn Leu Ala Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Ala Phe Thr Leu Arg Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln His 85 90 95 Leu Glu Tyr Pro Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys 100 105 110 <210> 43 <211> 475 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 43 Met Gly Arg Leu Thr Ser Ser Phe Leu Leu Leu Ile Val Pro Ala Tyr 1 5 10 15 Val Leu Ser Gln Val Thr Leu Lys Glu Ser Gly Pro Gly Ile Leu Arg 20 25 30 Pro Ser Gln Thr Leu Ser Leu Thr Cys Ser Phe Ser Gly Phe Ser Leu 35 40 45 Ser Thr Phe Gly Leu Ser Val Gly Trp Ile Arg Gln Pro Ser Gly Lys 50 55 60 Gly Leu Glu Trp Leu Ala His Ile Trp Trp Asp Asp Asp Lys Tyr Tyr 65 70 75 80 Asn Pro Ala Leu Lys Ser Arg Leu Thr Ile Ser Lys Asp Thr Ser Lys 85 90 95 Asn Gln Val Phe Leu Lys Ile Ala Asn Val Asp Thr Ala Asp Thr Ala 100 105 110 Thr Tyr Tyr Cys Ala Arg Ile Gly Ala Asp Ala Leu Pro Phe Asp Tyr 115 120 125 Trp Gly Gln Gly Thr Thr Leu Thr Val Ser Ser Ala Lys Thr Thr Pro 130 135 140 Pro Ser Val Tyr Pro Leu Ala Pro Gly Cys Gly Asp Thr Thr Gly Ser 145 150 155 160 Ser Val Thr Ser Gly Cys Leu Val Lys Gly Tyr Phe Pro Glu Pro Val 165 170 175 Thr Val Thr Trp Asn Ser Gly Ser Leu Ser Ser Ser Val His Thr Phe 180 185 190 Pro Ala Leu Leu Gln Ser Gly Leu Tyr Thr Met Ser Ser Ser Val Thr 195 200 205 Val Pro Ser Ser Thr Trp Pro Ser Gln Thr Val Thr Cys Ser Val Ala 210 215 220 His Pro Ala Ser Ser Thr Thr Thr Val Asp Lys Lys Leu Glu Pro Ser Gly 225 230 235 240 Pro Ile Ser Thr Ile Asn Pro Cys Pro Pro Cys Lys Glu Cys His Lys 245 250 255 Cys Pro Ala Pro Asn Leu Glu Gly Gly Pro Ser Val Phe Ile Phe Pro 260 265 270 Pro Asn Ile Lys Asp Val Leu Met Ile Ser Leu Thr Pro Lys Val Thr 275 280 285 Cys Val Val Val Asp Val Ser Glu Asp Asp Pro Asp Val Gln Ile Ser 290 295 300 Trp Phe Val Asn Asn Val Glu Val His Thr Ala Gln Thr Gln Thr His 305 310 315 320 Arg Glu Asp Tyr Asn Ser Thr Ile Arg Val Val Ser Thr Leu Pro Ile 325 330 335 Gln His Gln Asp Trp Met Ser Gly Lys Glu Phe Lys Cys Lys Val Asn 340 345 350 Asn Lys Asp Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ile Lys 355 360 365 Gly Leu Val Arg Ala Pro Gln Val Tyr Thr Leu Pro Pro Pro Ala Glu 370 375 380 Gln Leu Ser Arg Lys Asp Val Ser Leu Thr Cys Leu Val Val Gly Phe 385 390 395 400 Asn Pro Gly Asp Ile Ser Val Glu Trp Thr Ser Asn Gly His Thr Glu 405 410 415 Glu Asn Tyr Lys Asp Thr Ala Pro Val Leu Asp Ser Asp Gly Ser Tyr 420 425 430 Phe Ile Tyr Ser Lys Leu Asn Met Lys Thr Ser Lys Trp Glu Lys Thr 435 440 445 Asp Ser Phe Ser Cys Asn Val Arg His Glu Gly Leu Lys Asn Tyr Tyr 450 455 460 Leu Lys Lys Thr Ile Ser Arg Ser Pro Gly Lys 465 470 475 <210> 44 <211> 239 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 44 Met Arg Cys Leu Ala Glu Phe Leu Gly Leu Leu Val Leu Trp Ile Pro 1 5 10 15 Gly Ala Ile Gly Asp Ile Val Leu Thr Gln Thr Ala Pro Ser Val Pro 20 25 30 Val Thr Pro Gly Glu Ser Val Ser Ile Ser Cys Arg Ser Ser Lys Ser 35 40 45 Leu Leu His Ser Asn Gly Asn Thr Tyr Leu Tyr Trp Phe Leu Gln Arg 50 55 60 Pro Gly Gln Ser Pro Gln Leu Leu Ile Tyr Arg Met Ser Asn Leu Ala 65 70 75 80 Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Ala Phe 85 90 95 Thr Leu Arg Ile Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr 100 105 110 Cys Met Gln His Leu Glu Tyr Pro Phe Thr Phe Gly Ser Gly Thr Lys 115 120 125 Leu Glu Ile Lys Arg Ala Asp Ala Ala Pro Thr Val Ser Ile Phe Pro 130 135 140 Pro Ser Ser Glu Gln Leu Thr Ser Gly Gly Ala Ser Val Val Cys Phe 145 150 155 160 Leu Asn Asn Phe Tyr Pro Arg Asp Ile Asn Val Lys Trp Lys Ile Asp 165 170 175 Gly Ser Glu Arg Gln Asn Gly Val Leu Asn Ser Trp Thr Asp Gln Asp 180 185 190 Ser Lys Asp Ser Thr Tyr Ser Met Ser Ser Thr Leu Thr Leu Thr Lys 195 200 205 Asp Glu Tyr Glu Arg His Asn Ser Tyr Thr Cys Glu Ala Thr His Lys 210 215 220 Thr Ser Thr Ser Pro Ile Val Lys Ser Phe Asn Arg Asn Glu Cys 225 230 235 <210> 45 <211> 5 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 45 Asp His Ile Ile His 1 5 <210> 46 <211> 17 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 46 Tyr Ile Tyr Pro Arg Asp Gly Tyr Ile Lys Tyr Asn Glu Lys Phe Lys 1 5 10 15 Gly <210> 47 <211> 8 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 47 Gly Tyr Tyr Tyr Ala Met Asp Tyr 1 5 <210> 48 <211> 16 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 48 Arg Ser Ser Gln Ser Ile Val His Ser Ile Gly Asn Thr Tyr Leu Glu 1 5 10 15 <210> 49 <211> 9 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 49 Phe Gln Gly Ser His Val Pro Phe Thr 1 5 <210> 50 <211> 117 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 50 Gln Val Gln Leu Gln Gln Ser Asp Ala Glu Leu Val Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Ile Ser Cys Lys Val Ser Gly Tyr Thr Phe Thr Asp His 20 25 30 Ile Ile His Trp Met Lys Gln Arg Pro Glu Gln Gly Leu Glu Trp Ile 35 40 45 Gly Tyr Ile Tyr Pro Arg Asp Gly Tyr Ile Lys Tyr Asn Glu Lys Phe 50 55 60 Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Gln Val Asn Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys 85 90 95 Ala Arg Gly Tyr Tyr Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr Ser 100 105 110 Val Thr Val Ser Ser 115 <210> 51 <211> 112 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 51 Asp Val Leu Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly 1 5 10 15 Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser 20 25 30 Ile Gly Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Glu Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys Phe Gln Gly 85 90 95 Ser His Val Pro Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys 100 105 110 <210> 52 <211> 460 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 52 Met Glu Trp Ser Trp Val Ser Leu Phe Phe Leu Ser Val Thr Thr Gly 1 5 10 15 Val His Ser Gln Val Gln Leu Gln Gln Ser Asp Ala Glu Leu Val Lys 20 25 30 Pro Gly Ala Ser Val Lys Ile Ser Cys Lys Val Ser Gly Tyr Thr Phe 35 40 45 Thr Asp His Ile Ile His Trp Met Lys Gln Arg Pro Glu Gln Gly Leu 50 55 60 Glu Trp Ile Gly Tyr Ile Tyr Pro Arg Asp Gly Tyr Ile Lys Tyr Asn 65 70 75 80 Glu Lys Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser 85 90 95 Thr Ala Tyr Met Gln Val Asn Ser Leu Thr Ser Glu Asp Ser Ala Val 100 105 110 Tyr Phe Cys Ala Arg Gly Tyr Tyr Tyr Ala Met Asp Tyr Trp Gly Gln 115 120 125 Gly Thr Ser Val Thr Val Ser Ser Ala Lys Thr Thr Pro Pro Ser Val 130 135 140 Tyr Pro Leu Ala Pro Gly Ser Ala Ala Gln Thr Asn Ser Met Val Thr 145 150 155 160 Leu Gly Cys Leu Val Lys Gly Tyr Phe Pro Glu Pro Val Thr Val Thr 165 170 175 Trp Asn Ser Gly Ser Leu Ser Ser Gly Val His Thr Phe Pro Ala Val 180 185 190 Leu Gln Ser Asp Leu Tyr Thr Leu Ser Ser Ser Val Thr Val Pro Ser 195 200 205 Ser Thr Trp Pro Ser Gln Thr Val Thr Cys Asn Val Ala His Pro Ala 210 215 220 Ser Ser Thr Lys Val Asp Lys Lys Ile Val Pro Arg Asp Cys Gly Cys 225 230 235 240 Lys Pro Cys Ile Cys Thr Val Pro Glu Val Ser Ser Val Phe Ile Phe 245 250 255 Pro Pro Lys Pro Lys Asp Val Leu Thr Ile Thr Leu Thr Pro Lys Val 260 265 270 Thr Cys Val Val Val Asp Ile Ser Lys Asp Asp Pro Glu Val Gln Phe 275 280 285 Ser Trp Phe Val Asp Asp Val Glu Val His Thr Ala Gln Thr Gln Pro 290 295 300 Arg Glu Glu Gln Phe Asn Ser Thr Phe Arg Ser Val Ser Glu Leu Pro 305 310 315 320 Ile Met His Gln Asp Trp Leu Asn Gly Lys Glu Phe Lys Cys Arg Val 325 330 335 Asn Ser Ala Ala Phe Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr 340 345 350 Lys Gly Arg Pro Lys Ala Pro Gln Val Tyr Thr Ile Pro Pro Pro Lys 355 360 365 Glu Gln Met Ala Lys Asp Lys Val Ser Leu Thr Cys Met Ile Thr Asp 370 375 380 Phe Phe Pro Glu Asp Ile Thr Val Glu Trp Gln Trp Asn Gly Gln Pro 385 390 395 400 Ala Glu Asn Tyr Lys Asn Thr Gln Pro Ile Met Asp Thr Asp Gly Ser 405 410 415 Tyr Phe Val Tyr Ser Lys Leu Asn Val Gln Lys Ser Asn Trp Glu Ala 420 425 430 Gly Asn Thr Phe Thr Cys Ser Val Leu His Glu Gly Leu His Asn His 435 440 445 His Thr Glu Lys Ser Leu Ser His Ser Pro Gly Lys 450 455 460 <210> 53 <211> 238 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 53 Met Lys Leu Pro Val Arg Leu Leu Val Leu Met Phe Trp Ile Pro Ala 1 5 10 15 Ser Arg Ser Asp Val Leu Met Thr Gln Thr Pro Leu Ser Leu Pro Val 20 25 30 Ser Leu Gly Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile 35 40 45 Val His Ser Ile Gly Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro 50 55 60 Gly Gln Ser Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser 65 70 75 80 Gly Val Pro Glu Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr 85 90 95 Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys 100 105 110 Phe Gln Gly Ser His Val Pro Phe Thr Phe Gly Ser Gly Thr Lys Leu 115 120 125 Glu Ile Lys Arg Ala Asp Ala Ala Pro Thr Val Ser Ile Phe Pro Pro 130 135 140 Ser Ser Glu Gln Leu Thr Ser Gly Gly Ala Ser Val Val Cys Phe Leu 145 150 155 160 Asn Asn Phe Tyr Pro Lys Asp Ile Asn Val Lys Trp Lys Ile Asp Gly 165 170 175 Ser Glu Arg Gln Asn Gly Val Leu Asn Ser Trp Thr Asp Gln Asp Ser 180 185 190 Lys Asp Ser Thr Tyr Ser Met Ser Ser Thr Leu Thr Leu Thr Lys Asp 195 200 205 Glu Tyr Glu Arg His Asn Ser Tyr Thr Cys Glu Ala Thr His Lys Thr 210 215 220 Ser Thr Ser Pro Ile Val Lys Ser Phe Asn Arg Asn Glu Cys 225 230 235 <210> 54 <211> 5 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 54 Ser Tyr Trp Met His 1 5 <210> 55 <211> 17 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 55 Met Ile Asp Pro Ser Asp Val Tyr Thr Asn Tyr Asn Pro Lys Phe Lys 1 5 10 15 Gly <210> 56 <211> 6 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 56 Asn Tyr Ser Gly Asp Tyr 1 5 <210> 57 <211> 115 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 57 Gln Val Gln Leu Leu Gln Pro Gly Ala Glu Leu Val Arg Pro Gly Thr 1 5 10 15 Ser Val Lys Leu Ser Cys Lys Thr Ser Gly Tyr Thr Phe Ser Ser Tyr 20 25 30 Trp Met His Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Met Ile Asp Pro Ser Asp Val Tyr Thr Asn Tyr Asn Pro Lys Phe 50 55 60 Lys Gly Lys Ala Thr Leu Thr Val Asp Thr Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Asn Tyr Ser Gly Asp Tyr Trp Gly Gln Gly Thr Thr Leu Thr 100 105 110 Val Ser Ser 115 <210> 58 <211> 112 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 58 Asp Val Leu Met Thr Gln Ile Pro Leu Ser Leu Pro Val Ser Leu Gly 1 5 10 15 Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser 20 25 30 Asn Gly Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys Phe Gln Gly 85 90 95 Ser Tyr Val Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 <210> 59 <211> 458 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 59 Met Gly Trp Ser Cys Ile Ile Val Leu Leu Val Ser Thr Ala Thr Cys 1 5 10 15 Val His Ser Gln Val Gln Leu Leu Gln Pro Gly Ala Glu Leu Val Arg 20 25 30 Pro Gly Thr Ser Val Lys Leu Ser Cys Lys Thr Ser Gly Tyr Thr Phe 35 40 45 Ser Ser Tyr Trp Met His Trp Val Lys Gln Arg Pro Gly Gln Gly Leu 50 55 60 Glu Trp Ile Gly Met Ile Asp Pro Ser Asp Val Tyr Thr Asn Tyr Asn 65 70 75 80 Pro Lys Phe Lys Gly Lys Ala Thr Leu Thr Val Asp Thr Ser Ser Ser 85 90 95 Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val 100 105 110 Tyr Tyr Cys Ala Arg Asn Tyr Ser Gly Asp Tyr Trp Gly Gln Gly Thr 115 120 125 Thr Leu Thr Val Ser Ser Ala Lys Thr Thr Pro Pro Ser Val Tyr Pro 130 135 140 Leu Ala Pro Gly Ser Ala Ala Gln Thr Asn Ser Met Val Thr Leu Gly 145 150 155 160 Cys Leu Val Lys Gly Tyr Phe Pro Glu Pro Val Thr Val Thr Trp Asn 165 170 175 Ser Gly Ser Leu Ser Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 180 185 190 Ser Asp Leu Tyr Thr Leu Ser Ser Ser Ser Val Thr Val Pro Ser Ser Thr 195 200 205 Trp Pro Ser Gln Thr Val Thr Cys Asn Val Ala His Pro Ala Ser Ser 210 215 220 Thr Lys Val Asp Lys Lys Ile Val Pro Arg Asp Cys Gly Cys Lys Pro 225 230 235 240 Cys Ile Cys Thr Val Pro Glu Val Ser Ser Val Phe Ile Phe Pro Pro 245 250 255 Lys Pro Lys Asp Val Leu Thr Ile Thr Leu Thr Pro Lys Val Thr Cys 260 265 270 Val Val Val Asp Ile Ser Lys Asp Asp Pro Glu Val Gln Phe Ser Trp 275 280 285 Phe Val Asp Asp Val Glu Val His Thr Ala Gln Thr Gln Pro Arg Glu 290 295 300 Glu Gln Phe Asn Ser Thr Phe Arg Ser Val Ser Glu Leu Pro Ile Met 305 310 315 320 His Gln Asp Trp Leu Asn Gly Lys Glu Phe Lys Cys Arg Val Asn Ser 325 330 335 Ala Ala Phe Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr Lys Gly 340 345 350 Arg Pro Lys Ala Pro Gln Val Tyr Thr Ile Pro Pro Pro Lys Glu Gln 355 360 365 Met Ala Lys Asp Lys Val Ser Leu Thr Cys Met Ile Thr Asp Phe Phe 370 375 380 Pro Glu Asp Ile Thr Val Glu Trp Gln Trp Asn Gly Gln Pro Ala Glu 385 390 395 400 Asn Tyr Lys Asn Thr Gln Pro Ile Met Asp Thr Asp Gly Ser Tyr Phe 405 410 415 Val Tyr Ser Lys Leu Asn Val Gln Lys Ser Asn Trp Glu Ala Gly Asn 420 425 430 Thr Phe Thr Cys Ser Val Leu His Glu Gly Leu His Asn His His Thr 435 440 445 Glu Lys Ser Leu Ser His Ser Pro Gly Lys 450 455 <210> 60 <211> 238 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 60 Met Lys Leu Pro Val Arg Leu Leu Val Leu Met Phe Trp Ile Pro Ala 1 5 10 15 Ser Ser Ser Asp Val Leu Met Thr Gln Ile Pro Leu Ser Leu Pro Val 20 25 30 Ser Leu Gly Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile 35 40 45 Val His Ser Asn Gly Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro 50 55 60 Gly Gln Ser Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser 65 70 75 80 Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr 85 90 95 Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys 100 105 110 Phe Gln Gly Ser Tyr Val Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu 115 120 125 Glu Ile Lys Arg Ala Asp Ala Ala Pro Thr Val Ser Ile Phe Pro Pro 130 135 140 Ser Ser Glu Gln Leu Thr Ser Gly Gly Ala Ser Val Val Cys Phe Leu 145 150 155 160 Asn Asn Phe Tyr Pro Arg Asp Ile Asn Val Lys Trp Lys Ile Asp Gly 165 170 175 Ser Glu Arg Gln Asn Gly Val Leu Asn Ser Trp Thr Asp Gln Asp Ser 180 185 190 Lys Asp Ser Thr Tyr Ser Met Ser Ser Thr Leu Thr Leu Thr Lys Asp 195 200 205 Glu Tyr Glu Arg His Asn Ser Tyr Thr Cys Glu Ala Thr His Lys Thr 210 215 220 Ser Thr Ser Pro Ile Val Lys Ser Phe Asn Arg Asn Glu Cys 225 230 235 <210> 61 <211> 5 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 61 Thr Tyr Gly Met Ser 1 5 <210> 62 <211> 17 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 62 Trp Ile Asn Thr Tyr Ser Gly Val Pro Thr Tyr Ala Asp Asp Phe Lys 1 5 10 15 Gly <210> 63 <211> 12 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 63 Gly Arg Asp Gly Tyr Gln Val Ala Trp Phe Ala Tyr 1 5 10 <210> 64 <211> 11 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 64 Ile Thr Ser Thr Asp Ile Asp Asp Asp Met Asn 1 5 10 <210> 65 <211> 7 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 65 Glu Gly Asn Thr Leu Arg Pro 1 5 <210> 66 <211> 9 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 66 Leu Gln Ser Asp Asn Leu Pro Tyr Thr 1 5 <210> 67 <211> 121 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 67 Gln Ile Gln Leu Val Gln Ser Gly Pro Glu Leu Lys Lys Pro Gly Glu 1 5 10 15 Ala Val Lys Ile Ser Cys Lys Ser Ser Gly Tyr Thr Phe Thr Thr Tyr 20 25 30 Gly Met Ser Trp Val Lys Gln Ala Pro Gly Arg Ala Leu Lys Trp Met 35 40 45 Gly Trp Ile Asn Thr Tyr Ser Gly Val Pro Thr Tyr Ala Asp Asp Phe 50 55 60 Lys Gly Arg Phe Ala Phe Ser Leu Glu Ser Ser Ala Ser Thr Ala Tyr 65 70 75 80 Leu Gln Ile Asn Asn Leu Lys Asn Glu Asp Thr Ala Thr Tyr Phe Cys 85 90 95 Ala Arg Gly Arg Asp Gly Tyr Gln Val Ala Trp Phe Ala Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ala 115 120 <210> 68 <211> 107 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 68 Glu Thr Thr Val Thr Gln Ser Pro Ala Ser Leu Ser Met Ala Ile Gly 1 5 10 15 Asp Lys Val Thr Ile Arg Cys Ile Thr Ser Thr Asp Ile Asp Asp Asp 20 25 30 Met Asn Trp Phe Gln Gln Lys Pro Gly Glu Pro Pro Lys Leu Leu Ile 35 40 45 Ser Glu Gly Asn Thr Leu Arg Pro Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Tyr Gly Thr Asp Phe Ile Phe Thr Ile Glu Asn Met Leu Ser 65 70 75 80 Glu Asp Val Ala Asp Tyr Tyr Cys Leu Gln Ser Asp Asn Leu Pro Tyr 85 90 95 Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 69 <211> 464 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 69 Met Gly Trp Leu Trp Asn Leu Leu Phe Leu Met Ala Ala Ala Gln Ser 1 5 10 15 Ala Gln Ala Gln Ile Gln Leu Val Gln Ser Gly Pro Glu Leu Lys Lys 20 25 30 Pro Gly Glu Ala Val Lys Ile Ser Cys Lys Ser Ser Gly Tyr Thr Phe 35 40 45 Thr Thr Tyr Gly Met Ser Trp Val Lys Gln Ala Pro Gly Arg Ala Leu 50 55 60 Lys Trp Met Gly Trp Ile Asn Thr Tyr Ser Gly Val Pro Thr Tyr Ala 65 70 75 80 Asp Asp Phe Lys Gly Arg Phe Ala Phe Ser Leu Glu Ser Ser Ala Ser 85 90 95 Thr Ala Tyr Leu Gln Ile Asn Asn Leu Lys Asn Glu Asp Thr Ala Thr 100 105 110 Tyr Phe Cys Ala Arg Gly Arg Asp Gly Tyr Gln Val Ala Trp Phe Ala 115 120 125 Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ala Ala Lys Thr Thr 130 135 140 Pro Pro Ser Val Tyr Pro Leu Ala Pro Gly Ser Ala Ala Gln Thr Asn 145 150 155 160 Ser Met Val Thr Leu Gly Cys Leu Val Lys Gly Tyr Phe Pro Glu Pro 165 170 175 Val Thr Val Thr Trp Asn Ser Gly Ser Leu Ser Ser Gly Val His Thr 180 185 190 Phe Pro Ala Val Leu Gln Ser Asp Leu Tyr Thr Leu Ser Ser Ser Val 195 200 205 Thr Val Pro Ser Ser Thr Trp Pro Ser Gln Thr Val Thr Cys Asn Val 210 215 220 Ala His Pro Ala Ser Ser Thr Lys Val Asp Lys Lys Ile Val Pro Arg 225 230 235 240 Asp Cys Gly Cys Lys Pro Cys Ile Cys Thr Val Pro Glu Val Ser Ser 245 250 255 Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Val Leu Thr Ile Thr Leu 260 265 270 Thr Pro Lys Val Thr Cys Val Val Val Asp Ile Ser Lys Asp Asp Pro 275 280 285 Glu Val Gln Phe Ser Trp Phe Val Asp Asp Val Glu Val His Thr Ala 290 295 300 Gln Thr Gln Pro Arg Glu Glu Gln Phe Asn Ser Thr Phe Arg Ser Val 305 310 315 320 Ser Glu Leu Pro Ile Met His Gln Asp Trp Leu Asn Gly Lys Glu Phe 325 330 335 Lys Cys Arg Val Asn Ser Ala Ala Phe Pro Ala Pro Ile Glu Lys Thr 340 345 350 Ile Ser Lys Thr Lys Gly Arg Pro Lys Ala Pro Gln Val Tyr Thr Ile 355 360 365 Pro Pro Pro Lys Glu Gln Met Ala Lys Asp Lys Val Ser Leu Thr Cys 370 375 380 Met Ile Thr Asp Phe Phe Pro Glu Asp Ile Thr Val Glu Trp Gln Trp 385 390 395 400 Asn Gly Gln Pro Ala Glu Asn Tyr Lys Asn Thr Gln Pro Ile Met Asp 405 410 415 Thr Asp Gly Ser Tyr Phe Val Tyr Ser Lys Leu Asn Val Gln Lys Ser 420 425 430 Asn Trp Glu Ala Gly Asn Thr Phe Thr Cys Ser Val Leu His Glu Gly 435 440 445 Leu His Asn His His Thr Glu Lys Ser Leu Ser His Ser Pro Gly Lys 450 455 460 <210> 70 <211> 234 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 70 Met Phe Ser Leu Ala Leu Leu Leu Ser Leu Leu Leu Leu Cys Val Ser 1 5 10 15 Asp Ser Arg Ala Glu Thr Thr Thr Val Thr Gln Ser Pro Ala Ser Leu Ser 20 25 30 Met Ala Ile Gly Asp Lys Val Thr Ile Arg Cys Ile Thr Ser Thr Asp 35 40 45 Ile Asp Asp Asp Met Asn Trp Phe Gln Gln Lys Pro Gly Glu Pro Pro 50 55 60 Lys Leu Leu Ile Ser Glu Gly Asn Thr Leu Arg Pro Gly Val Pro Ser 65 70 75 80 Arg Phe Ser Gly Ser Gly Tyr Gly Thr Asp Phe Ile Phe Thr Ile Glu 85 90 95 Asn Met Leu Ser Glu Asp Val Ala Asp Tyr Tyr Cys Leu Gln Ser Asp 100 105 110 Asn Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg 115 120 125 Ala Asp Ala Ala Pro Thr Val Ser Ile Phe Pro Pro Ser Ser Glu Gln 130 135 140 Leu Thr Ser Gly Gly Ala Ser Val Val Cys Phe Leu Asn Asn Phe Tyr 145 150 155 160 Pro Arg Asp Ile Asn Val Lys Trp Lys Ile Asp Gly Ser Glu Arg Gln 165 170 175 Asn Gly Val Leu Asn Ser Trp Thr Asp Gln Asp Ser Lys Asp Ser Thr 180 185 190 Tyr Ser Met Ser Ser Thr Leu Thr Leu Thr Lys Asp Glu Tyr Glu Arg 195 200 205 His Asn Ser Tyr Thr Cys Glu Ala Thr His Lys Thr Ser Thr Ser Pro 210 215 220 Ile Val Lys Ser Phe Asn Arg Asn Glu Cys 225 230 <210> 71 <211> 5 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 71 Asn Tyr Trp Met His 1 5 <210> 72 <211> 17 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 72 Met Ile Asp Pro Ser Asp Ser Tyr Thr Asn Tyr Asn Pro Lys Phe Lys 1 5 10 15 Gly <210> 73 <211> 115 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 73 Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Arg Pro Gly Thr 1 5 10 15 Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr 20 25 30 Trp Met His Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Met Ile Asp Pro Ser Asp Ser Tyr Thr Asn Tyr Asn Pro Lys Phe 50 55 60 Lys Gly Lys Ala Thr Leu Thr Val Asp Thr Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Asn Tyr Ser Gly Asp Tyr Trp Gly Gln Gly Thr Thr Leu Thr 100 105 110 Val Ser Ser 115 <210> 74 <211> 112 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 74 Asp Val Leu Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly 1 5 10 15 Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser 20 25 30 Asn Gly Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys Phe Gln Gly 85 90 95 Ser Tyr Val Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 <210> 75 <211> 458 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 75 Met Gly Trp Ser Cys Ile Ile Val Leu Leu Val Ser Thr Ala Thr Gly 1 5 10 15 Val His Ser Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Arg 20 25 30 Pro Gly Thr Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe 35 40 45 Thr Asn Tyr Trp Met His Trp Val Lys Gln Arg Pro Gly Gln Gly Leu 50 55 60 Glu Trp Ile Gly Met Ile Asp Pro Ser Asp Ser Tyr Thr Asn Tyr Asn 65 70 75 80 Pro Lys Phe Lys Gly Lys Ala Thr Leu Thr Val Asp Thr Ser Ser Ser 85 90 95 Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val 100 105 110 Tyr Tyr Cys Ala Arg Asn Tyr Ser Gly Asp Tyr Trp Gly Gln Gly Thr 115 120 125 Thr Leu Thr Val Ser Ser Ala Lys Thr Thr Pro Pro Ser Val Tyr Pro 130 135 140 Leu Ala Pro Gly Ser Ala Ala Gln Thr Asn Ser Met Val Thr Leu Gly 145 150 155 160 Cys Leu Val Lys Gly Tyr Phe Pro Glu Pro Val Thr Val Thr Trp Asn 165 170 175 Ser Gly Ser Leu Ser Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 180 185 190 Ser Asp Leu Tyr Thr Leu Ser Ser Ser Ser Val Thr Val Pro Ser Ser Thr 195 200 205 Trp Pro Ser Gln Thr Val Thr Cys Asn Val Ala His Pro Ala Ser Ser 210 215 220 Thr Lys Val Asp Lys Lys Ile Val Pro Arg Asp Cys Gly Cys Lys Pro 225 230 235 240 Cys Ile Cys Thr Val Pro Glu Val Ser Ser Val Phe Ile Phe Pro Pro 245 250 255 Lys Pro Lys Asp Val Leu Thr Ile Thr Leu Thr Pro Lys Val Thr Cys 260 265 270 Val Val Val Asp Ile Ser Lys Asp Asp Pro Glu Val Gln Phe Ser Trp 275 280 285 Phe Val Asp Asp Val Glu Val His Thr Ala Gln Thr Gln Pro Arg Glu 290 295 300 Glu Gln Phe Asn Ser Thr Phe Arg Ser Val Ser Glu Leu Pro Ile Met 305 310 315 320 His Gln Asp Trp Leu Asn Gly Lys Glu Phe Lys Cys Arg Val Asn Ser 325 330 335 Ala Ala Phe Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr Lys Gly 340 345 350 Arg Pro Lys Ala Pro Gln Val Tyr Thr Ile Pro Pro Pro Lys Glu Gln 355 360 365 Met Ala Lys Asp Lys Val Ser Leu Thr Cys Met Ile Thr Asp Phe Phe 370 375 380 Pro Glu Asp Ile Thr Val Glu Trp Gln Trp Asn Gly Gln Pro Ala Glu 385 390 395 400 Asn Tyr Lys Asn Thr Gln Pro Ile Met Asp Thr Asp Gly Ser Tyr Phe 405 410 415 Val Tyr Ser Lys Leu Asn Val Gln Lys Ser Asn Trp Glu Ala Gly Asn 420 425 430 Thr Phe Thr Cys Ser Val Leu His Glu Gly Leu His Asn His His Thr 435 440 445 Glu Lys Ser Leu Ser His Ser Pro Gly Lys 450 455 <210> 76 <211> 238 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 76 Met Lys Leu Pro Val Arg Leu Leu Val Leu Met Phe Trp Ile Pro Ala 1 5 10 15 Ser Ser Ser Asp Val Leu Met Thr Gln Thr Pro Leu Ser Leu Pro Val 20 25 30 Ser Leu Gly Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile 35 40 45 Val His Ser Asn Gly Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro 50 55 60 Gly Gln Ser Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser 65 70 75 80 Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr 85 90 95 Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys 100 105 110 Phe Gln Gly Ser Tyr Val Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu 115 120 125 Glu Ile Lys Arg Ala Asp Ala Ala Pro Thr Val Ser Ile Phe Pro Pro 130 135 140 Ser Ser Glu Gln Leu Thr Ser Gly Gly Ala Ser Val Val Cys Phe Leu 145 150 155 160 Asn Asn Phe Tyr Pro Arg Asp Ile Asn Val Lys Trp Lys Ile Asp Gly 165 170 175 Ser Glu Arg Gln Asn Gly Val Leu Asn Ser Trp Thr Asp Gln Asp Ser 180 185 190 Lys Asp Ser Thr Tyr Ser Met Ser Ser Thr Leu Thr Leu Thr Lys Asp 195 200 205 Glu Tyr Glu Arg His Asn Ser Tyr Thr Cys Glu Ala Thr His Lys Thr 210 215 220 Ser Thr Ser Pro Ile Val Lys Ser Phe Asn Arg Asn Glu Cys 225 230 235 <210> 77 <211> 449 <212> PRT <213> artificial sequence <220> 223 <#223> <400> 77 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Ala Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Thr Ile Ser Asp Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Trp Gly Asp Tyr Asp Gly Phe Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125 Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140 Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160 Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175 Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190 Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro 195 200 205 Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220 Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro 225 230 235 240 Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser 245 250 255 Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 260 265 270 Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285 Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val 290 295 300 Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu 305 310 315 320 Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys 325 330 335 Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 340 345 350 Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr 355 360 365 Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 370 375 380 Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 385 390 395 400 Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys 405 410 415 Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430 Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445 Lys <210> 78 <211> 214 <212> PRT <213> artificial sequence <220> <223> Antibody light chain <400> 78 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Glu Ile Ser Gly Tyr 20 25 30 Leu Ser Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Arg Leu Ile 35 40 45 Tyr Ala Ala Ser Thr Leu Asp Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln Tyr Asp Ser Tyr Pro Tyr 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 <210> 79 <211> 119 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 79 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Thr Ile Ser Asp Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Asn Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Trp Gly Asp Tyr Asp Gly Phe Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 80 <211> 119 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 80 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Ala Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Thr Ile Ser Asp Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Asn Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Trp Gly Asp Tyr Asp Gly Phe Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 81 <211> 119 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 81 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Ala Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Thr Ile Ser Asp Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Trp Gly Asp Tyr Asp Gly Phe Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 82 <211> 119 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 82 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Thr Ile Ser Asp Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Asn Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Trp Gly Asp Tyr Asp Gly Phe Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 83 <211> 119 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 83 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Thr Ile Ser Asp Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Asn Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Trp Gly Asp Tyr Asp Gly Phe Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 84 <211> 119 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 84 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Thr Ile Ser Asp Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Asn Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Ser Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Trp Gly Asp Tyr Asp Gly Phe Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 85 <211> 119 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 85 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Thr Ile Ser Asp Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Asn Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Trp Gly Asp Tyr Asp Gly Phe Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 <210> 86 <211> 107 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 86 Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Glu Ile Ser Gly Tyr 20 25 30 Leu Ser Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ala Ala Ser Thr Leu Asp Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln Tyr Asp Ser Tyr Pro Tyr 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 87 <211> 107 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 87 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Glu Ile Ser Gly Tyr 20 25 30 Leu Ser Trp Phe Gln Gln Lys Pro Gly Lys Ala Pro Lys Ser Leu Ile 35 40 45 Tyr Ala Ala Ser Thr Leu Asp Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln Tyr Asp Ser Tyr Pro Tyr 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 88 <211> 107 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 88 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Glu Ile Ser Gly Tyr 20 25 30 Leu Ser Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Arg Leu Ile 35 40 45 Tyr Ala Ala Ser Thr Leu Asp Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln Tyr Asp Ser Tyr Pro Tyr 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 89 <211> 107 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 89 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Glu Ile Ser Gly Tyr 20 25 30 Leu Ser Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ala Ala Ser Thr Leu Asp Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Ile Ala Thr Tyr Tyr Cys Leu Gln Tyr Asp Ser Tyr Pro Tyr 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 90 <211> 107 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 90 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Glu Ile Ser Gly Tyr 20 25 30 Leu Ser Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ala Ala Ser Thr Leu Asp Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln Tyr Asp Ser Tyr Pro Tyr 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 91 <211> 107 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 91 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Glu Ile Ser Gly Tyr 20 25 30 Leu Ser Trp Leu Gln Gln Lys Pro Gly Gly Ala Ile Lys Arg Leu Ile 35 40 45 Tyr Ala Ala Ser Thr Leu Asp Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Ser Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln Tyr Asp Ser Tyr Pro Tyr 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 92 <211> 468 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 92 Met Asn Phe Gly Leu Ser Leu Met Phe Leu Val Leu Val Leu Lys Gly 1 5 10 15 Val Gln Cys Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys 20 25 30 Pro Gly Gly Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe 35 40 45 Ser Asp Tyr Ala Met Ser Trp Val Arg Gln Thr Pro Glu Lys Arg Leu 50 55 60 Glu Trp Val Ala Thr Ile Ser Asp Gly Gly Thr Tyr Thr Tyr Tyr Pro 65 70 75 80 Asp Asn Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn 85 90 95 Asn Leu Tyr Leu Gln Met Ser His Leu Lys Ser Glu Asp Thr Ala Met 100 105 110 Tyr Tyr Cys Ala Arg Glu Trp Gly Asp Tyr Asp Gly Phe Asp Tyr Trp 115 120 125 Gly Gln Gly Thr Thr Leu Thr Val Ser Ser Ala Ser Thr Lys Gly Pro 130 135 140 Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr 145 150 155 160 Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr 165 170 175 Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro 180 185 190 Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr 195 200 205 Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn 210 215 220 His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser 225 230 235 240 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 245 250 255 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 260 265 270 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 275 280 285 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 290 295 300 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 305 310 315 320 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 325 330 335 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 340 345 350 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 355 360 365 Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val 370 375 380 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 385 390 395 400 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 405 410 415 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 420 425 430 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 435 440 445 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 450 455 460 Ser Pro Gly Lys 465 <210> 93 <211> 236 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 93 Met Asp Met Arg Val Pro Ala His Val Phe Gly Phe Leu Leu Leu Trp 1 5 10 15 Phe Pro Gly Thr Arg Cys Asp Ile Gln Met Thr Gln Ser Pro Ser Ser 20 25 30 Leu Ser Ala Ser Leu Gly Glu Arg Val Ser Leu Thr Cys Arg Ala Ser 35 40 45 Gln Glu Ile Ser Gly Tyr Leu Ser Trp Leu Gln Gln Lys Pro Asp Gly 50 55 60 Thr Ile Lys Arg Leu Ile Tyr Ala Ala Ser Thr Leu Asp Ser Gly Val 65 70 75 80 Pro Lys Arg Phe Ser Gly Ser Arg Ser Gly Ser Asp Tyr Ser Leu Thr 85 90 95 Ile Gly Ser Leu Glu Ser Glu Asp Leu Ala Asp Tyr Tyr Cys Leu Gln 100 105 110 Tyr Asp Ser Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile 115 120 125 Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp 130 135 140 Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn 145 150 155 160 Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu 165 170 175 Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp 180 185 190 Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr 195 200 205 Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser 210 215 220 Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 225 230 235 <210> 94 <211> 471 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 94 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Gln Val Gln Leu Val Glu Ser Gly Gly Gly 20 25 30 Leu Val Lys Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly 35 40 45 Phe Thr Phe Ser Asp Tyr Ala Met Ser Trp Ile Arg Gln Ala Pro Gly 50 55 60 Lys Gly Leu Glu Trp Val Ser Thr Ile Ser Asp Gly Gly Thr Tyr Thr 65 70 75 80 Tyr Tyr Pro Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn 85 90 95 Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp 100 105 110 Thr Ala Val Tyr Tyr Cys Ala Arg Glu Trp Gly Asp Tyr Asp Gly Phe 115 120 125 Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr 130 135 140 Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser 145 150 155 160 Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu 165 170 175 Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His 180 185 190 Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser 195 200 205 Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys 210 215 220 Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu 225 230 235 240 Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 245 250 255 Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 260 265 270 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 275 280 285 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 290 295 300 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 305 310 315 320 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 325 330 335 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 340 345 350 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 355 360 365 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys 370 375 380 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 385 390 395 400 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 405 410 415 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 420 425 430 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 435 440 445 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 450 455 460 Leu Ser Leu Ser Pro Gly Lys 465 470 <210> 95 <211> 467 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 95 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Gln Val Gln Leu Val Glu Ser Gly Gly Gly 20 25 30 Leu Val Lys Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly 35 40 45 Phe Thr Phe Ser Asp Tyr Ala Met Ser Trp Ile Arg Gln Ala Pro Gly 50 55 60 Lys Gly Leu Glu Trp Val Ser Thr Ile Ser Asp Gly Gly Thr Tyr Thr 65 70 75 80 Tyr Tyr Pro Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn 85 90 95 Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp 100 105 110 Thr Ala Val Tyr Tyr Cys Ala Arg Glu Trp Gly Asp Tyr Asp Gly Phe 115 120 125 Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr 130 135 140 Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser 145 150 155 160 Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu 165 170 175 Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His 180 185 190 Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser 195 200 205 Val Val Thr Val Pro Ser Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys 210 215 220 Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Thr Val Glu 225 230 235 240 Arg Lys Cys Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala 245 250 255 Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met 260 265 270 Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His 275 280 285 Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val 290 295 300 His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Phe 305 310 315 320 Arg Val Val Ser Val Leu Thr Val Val His Gln Asp Trp Leu Asn Gly 325 330 335 Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ala Pro Ile 340 345 350 Glu Lys Thr Ile Ser Lys Thr Lys Gly Gln Pro Arg Glu Pro Gln Val 355 360 365 Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser 370 375 380 Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu 385 390 395 400 Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro 405 410 415 Met Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val 420 425 430 Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met 435 440 445 His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser 450 455 460 Pro Gly Lys 465 <210> 96 <211> 236 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 96 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Asp Ile Gln Met Thr Gln Ser Pro Ser Ser 20 25 30 Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser 35 40 45 Gln Glu Ile Ser Gly Tyr Leu Ser Trp Phe Gln Gln Lys Pro Gly Lys 50 55 60 Ala Pro Lys Ser Leu Ile Tyr Ala Ala Ser Thr Leu Asp Ser Gly Val 65 70 75 80 Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 85 90 95 Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln 100 105 110 Tyr Asp Ser Tyr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile 115 120 125 Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp 130 135 140 Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn 145 150 155 160 Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu 165 170 175 Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp 180 185 190 Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr 195 200 205 Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser 210 215 220 Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 225 230 235 <210> 97 <211> 236 <212> PRT <213> artificial sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 97 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Arg Gly Ala Arg Cys Asp Ile Gln Met Thr Gln Ser Pro Ser Ser 20 25 30 Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser 35 40 45 Gln Glu Ile Ser Gly Tyr Leu Ser Trp Tyr Gln Gln Lys Pro Gly Lys 50 55 60 Ala Pro Lys Arg Leu Ile Tyr Ala Ala Ser Thr Leu Asp Ser Gly Val 65 70 75 80 Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr 85 90 95 Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln 100 105 110 Tyr Asp Ser Tyr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile 115 120 125 Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp 130 135 140 Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn 145 150 155 160 Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu 165 170 175 Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp 180 185 190 Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr 195 200 205 Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser 210 215 220 Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 225 230 235 <210> 98 <211> 449 <212> PRT <213> artificial sequence <220> <223> Antibody heavy chain no leader sequence <400> 98 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Ala Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Thr Ile Ser Asp Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Trp Gly Asp Tyr Asp Gly Phe Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125 Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140 Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160 Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175 Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190 Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro 195 200 205 Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys 210 215 220 Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro 225 230 235 240 Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser 245 250 255 Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 260 265 270 Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285 Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val 290 295 300 Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu 305 310 315 320 Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys 325 330 335 Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 340 345 350 Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr 355 360 365 Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 370 375 380 Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 385 390 395 400 Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys 405 410 415 Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430 Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445 Lys <210> 99 <211> 445 <212> PRT <213> artificial sequence <220> <223> Antibody heavy chain no leader sequence <400> 99 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr 20 25 30 Ala Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Thr Ile Ser Asp Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Trp Gly Asp Tyr Asp Gly Phe Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125 Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu 130 135 140 Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160 Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175 Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190 Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn Val Asp His Lys Pro 195 200 205 Ser Asn Thr Lys Val Asp Lys Thr Val Glu Arg Lys Cys Cys Val Glu 210 215 220 Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly Pro Ser Val Phe Leu 225 230 235 240 Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu 245 250 255 Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Gln 260 265 270 Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys 275 280 285 Pro Arg Glu Glu Gln Phe Asn Ser Thr Phe Arg Val Val Ser Val Leu 290 295 300 Thr Val Val His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys 305 310 315 320 Val Ser Asn Lys Gly Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys 325 330 335 Thr Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser 340 345 350 Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys 355 360 365 Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln 370 375 380 Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Met Leu Asp Ser Asp Gly 385 390 395 400 Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln 405 410 415 Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn 420 425 430 His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <210> 100 <211> 214 <212> PRT <213> artificial sequence <220> <223> Antibody light chain no leader sequence <400> 100 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Glu Ile Ser Gly Tyr 20 25 30 Leu Ser Trp Phe Gln Gln Lys Pro Gly Lys Ala Pro Lys Ser Leu Ile 35 40 45 Tyr Ala Ala Ser Thr Leu Asp Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln Tyr Asp Ser Tyr Pro Tyr 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 <210> 101 <211> 214 <212> PRT <213> artificial sequence <220> <223> Antibody light chain no leader sequence <400> 101 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Glu Ile Ser Gly Tyr 20 25 30 Leu Ser Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Arg Leu Ile 35 40 45 Tyr Ala Ala Ser Thr Leu Asp Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln Tyr Asp Ser Tyr Pro Tyr 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 <210> 102 <211> 450 <212> PRT <213> artificial sequence <220> 223 <#223> <400> 102 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Asn Ile Lys Asp Thr 20 25 30 Tyr Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Arg Ile Tyr Pro Thr Asn Gly Tyr Thr Arg Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn Thr Ala Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ser Arg Trp Gly Gly Asp Gly Phe Tyr Ala Met Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 210 215 220 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly 225 230 235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu 260 265 270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 305 310 315 320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350 Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu 355 360 365 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410 415 Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 420 425 430 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440 445 Gly Lys 450 <210> 103 <211> 214 <212> PRT <213> artificial sequence <220> <223> Antibody light chain <400> 103 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Val Asn Thr Ala 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ser Ala Ser Phe Leu Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Arg Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln His Tyr Thr Thr Pro Pro 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 <210> 104 <211> 448 <212> PRT <213> artificial sequence <220> 223 <#223> <400> 104 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Asp Tyr 20 25 30 Thr Met Asp Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Asp Val Asn Pro Asn Ser Gly Gly Ser Ile Tyr Asn Gln Arg Phe 50 55 60 Lys Gly Arg Phe Thr Leu Ser Val Asp Arg Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Asn Leu Gly Pro Ser Phe Tyr Phe Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125 Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140 Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160 Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175 Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190 Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro 195 200 205 Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220 Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro 225 230 235 240 Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser 245 250 255 Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 260 265 270 Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285 Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val 290 295 300 Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu 305 310 315 320 Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys 325 330 335 Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 340 345 350 Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr 355 360 365 Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 370 375 380 Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 385 390 395 400 Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys 405 410 415 Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430 Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445 <210> 105 <211> 214 <212> PRT <213> artificial sequence <220> <223> Antibody light chain <400> 105 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asp Val Ser Ile Gly 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ser Ala Ser Tyr Arg Tyr Thr Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Ile Tyr Pro Tyr 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 <210> 106 <211> 447 <212> PRT <213> artificial sequence <220> 223 <#223> <400> 106 Gln Val Gln Leu Gln Gln Trp Gly Ala Gly Leu Leu Lys Pro Ser Glu 1 5 10 15 Thr Leu Ser Leu Thr Cys Ala Val Tyr Gly Gly Ser Phe Ser Gly Tyr 20 25 30 Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45 Gly Glu Ile Asn His Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu Lys 50 55 60 Ser Arg Val Thr Ile Ser Val Glu Thr Ser Lys Asn Gln Phe Ser Leu 65 70 75 80 Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Arg Asp Lys Trp Thr Trp Tyr Phe Asp Leu Trp Gly Arg Gly Thr Leu 100 105 110 Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu 115 120 125 Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys 130 135 140 Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser 145 150 155 160 Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser 165 170 175 Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser 180 185 190 Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn 195 200 205 Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His 210 215 220 Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val 225 230 235 240 Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr 245 250 255 Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu 260 265 270 Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys 275 280 285 Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser 290 295 300 Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys 305 310 315 320 Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile 325 330 335 Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro 340 345 350 Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu 355 360 365 Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn 370 375 380 Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser 385 390 395 400 Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg 405 410 415 Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu 420 425 430 His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <210> 107 <211> 220 <212> PRT <213> artificial sequence <220> <223> Antibody light chain <400> 107 Asp Ile Glu Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Glu Arg Ala Thr Ile Asn Cys Arg Ser Ser Gln Ser Val Leu Tyr Ser 20 25 30 Ser Ser Asn Arg Asn Tyr Leu Ala Trp Tyr Gln Gln Asn Pro Gly Gln 35 40 45 Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val 50 55 60 Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 65 70 75 80 Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln 85 90 95 Tyr Tyr Ser Thr Pro Arg Thr Phe Gly Gln Gly Thr Lys Val Glu Ile 100 105 110 Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp 115 120 125 Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn 130 135 140 Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu 145 150 155 160 Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp 165 170 175 Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr 180 185 190 Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser 195 200 205 Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 210 215 220 <210> 108 <211> 445 <212> PRT <213> artificial sequence <220> 223 <#223> <400> 108 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser His Tyr 20 25 30 Val Met Ala Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Ser Ser Ser Gly Gly Trp Thr Leu Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Thr Arg Gly Leu Lys Met Ala Thr Ile Phe Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125 Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu 130 135 140 Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160 Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175 Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190 Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn Val Asp His Lys Pro 195 200 205 Ser Asn Thr Lys Val Asp Lys Thr Val Glu Arg Lys Cys Cys Val Glu 210 215 220 Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly Pro Ser Val Phe Leu 225 230 235 240 Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu 245 250 255 Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Gln 260 265 270 Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys 275 280 285 Pro Arg Glu Glu Gln Phe Asn Ser Thr Phe Arg Val Val Ser Val Leu 290 295 300 Thr Val Val His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys 305 310 315 320 Val Ser Asn Lys Gly Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys 325 330 335 Thr Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser 340 345 350 Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys 355 360 365 Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln 370 375 380 Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Met Leu Asp Ser Asp Gly 385 390 395 400 Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln 405 410 415 Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn 420 425 430 His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <210> 109 <211> 217 <212> PRT <213> artificial sequence <220> <223> Antibody light chain <400> 109 Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Ser Tyr 20 25 30 Asn Val Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Ile Ile Tyr Glu Val Ser Gln Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Thr Glu Asp Glu Ala Asp Tyr Tyr Cys Cys Ser Tyr Ala Gly Ser 85 90 95 Ser Ile Phe Val Ile Phe Gly Gly Gly Thr Lys Val Thr Val Leu Gly 100 105 110 Gln Pro Lys Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser Glu 115 120 125 Glu Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Val Ser Asp Phe 130 135 140 Tyr Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp Gly Ser Pro Val 145 150 155 160 Lys Val Gly Val Glu Thr Thr Lys Pro Ser Lys Gln Ser Asn Asn Lys 165 170 175 Tyr Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser 180 185 190 His Arg Ser Tyr Ser Cys Arg Val Thr His Glu Gly Ser Thr Val Glu 195 200 205 Lys Thr Val Ala Pro Ala Glu Cys Ser 210 215 <210> 110 <211> 449 <212> PRT <213> artificial sequence <220> 223 <#223> <400> 110 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Arg Ser Ser 20 25 30 Tyr Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Tyr Ala Gly Thr Gly Ser Pro Ser Tyr Asn Gln Lys Leu 50 55 60 Gln Gly Arg Val Thr Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg His Arg Asp Tyr Tyr Ser Asn Ser Leu Thr Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 210 215 220 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly 225 230 235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu 260 265 270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 305 310 315 320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350 Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu 355 360 365 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410 415 Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 420 425 430 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440 445 Gly <210> 111 <211> 220 <212> PRT <213> artificial sequence <220> <223> Antibody light chain <400> 111 Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 1 5 10 15 Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Val Leu Asn Ser 20 25 30 Gly Asn Gln Lys Asn Tyr Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln 35 40 45 Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val 50 55 60 Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 65 70 75 80 Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Ser 85 90 95 Asp Tyr Ser Tyr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile 100 105 110 Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp 115 120 125 Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn 130 135 140 Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu 145 150 155 160 Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp 165 170 175 Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr 180 185 190 Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser 195 200 205 Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 210 215 220 <210> 112 <211> 446 <212> PRT <213> artificial sequence <220> 223 <#223> <400> 112 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Asn Ser Gln Gly Lys Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Trp Gly Asp Glu Gly Phe Asp Ile Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu 115 120 125 Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys 130 135 140 Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser 145 150 155 160 Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser 165 170 175 Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser 180 185 190 Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn 195 200 205 Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His 210 215 220 Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val 225 230 235 240 Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr 245 250 255 Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu 260 265 270 Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys 275 280 285 Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser 290 295 300 Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys 305 310 315 320 Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile 325 330 335 Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro 340 345 350 Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu 355 360 365 Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn 370 375 380 Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser 385 390 395 400 Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg 405 410 415 Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu 420 425 430 His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445 <210> 113 <211> 214 <212> PRT <213> artificial sequence <220> <223> Antibody light chain <400> 113 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Asn Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Gly Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Ser Ser Phe Pro Thr 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 <210> 114 <211> 25 <212> DNA <213> artificial sequence <220> <223> <400> 114 cgtcacaggc aggacccact gccca 25 <210> 115 <211> 1342 <212> PRT <213> Homo sapiens <400> 115 Met Arg Ala Asn Asp Ala Leu Gln Val Leu Gly Leu Leu Phe Ser Leu 1 5 10 15 Ala Arg Gly Ser Glu Val Gly Asn Ser Gln Ala Val Cys Pro Gly Thr 20 25 30 Leu Asn Gly Leu Ser Val Thr Gly Asp Ala Glu Asn Gln Tyr Gln Thr 35 40 45 Leu Tyr Lys Leu Tyr Glu Arg Cys Glu Val Val Met Gly Asn Leu Glu 50 55 60 Ile Val Leu Thr Gly His Asn Ala Asp Leu Ser Phe Leu Gln Trp Ile 65 70 75 80 Arg Glu Val Thr Gly Tyr Val Leu Val Ala Met Asn Glu Phe Ser Thr 85 90 95 Leu Pro Leu Pro Asn Leu Arg Val Val Arg Gly Thr Gln Val Tyr Asp 100 105 110 Gly Lys Phe Ala Ile Phe Val Met Leu Asn Tyr Asn Thr Asn Ser Ser 115 120 125 His Ala Leu Arg Gln Leu Arg Leu Thr Gln Leu Thr Glu Ile Leu Ser 130 135 140 Gly Gly Val Tyr Ile Glu Lys Asn Asp Lys Leu Cys His Met Asp Thr 145 150 155 160 Ile Asp Trp Arg Asp Ile Val Arg Asp Arg Asp Ala Glu Ile Val Val 165 170 175 Lys Asp Asn Gly Arg Ser Cys Pro Pro Cys His Glu Val Cys Lys Gly 180 185 190 Arg Cys Trp Gly Pro Gly Ser Glu Asp Cys Gln Thr Leu Thr Lys Thr 195 200 205 Ile Cys Ala Pro Gln Cys Asn Gly His Cys Phe Gly Pro Asn Pro Asn 210 215 220 Gln Cys Cys His Asp Glu Cys Ala Gly Gly Cys Ser Gly Pro Gln Asp 225 230 235 240 Thr Asp Cys Phe Ala Cys Arg His Phe Asn Asp Ser Gly Ala Cys Val 245 250 255 Pro Arg Cys Pro Gln Pro Leu Val Tyr Asn Lys Leu Thr Phe Gln Leu 260 265 270 Glu Pro Asn Pro His Thr Lys Tyr Gln Tyr Gly Gly Val Cys Val Ala 275 280 285 Ser Cys Pro His Asn Phe Val Val Asp Gln Thr Ser Cys Val Arg Ala 290 295 300 Cys Pro Pro Asp Lys Met Glu Val Asp Lys Asn Gly Leu Lys Met Cys 305 310 315 320 Glu Pro Cys Gly Gly Leu Cys Pro Lys Ala Cys Glu Gly Thr Gly Ser 325 330 335 Gly Ser Arg Phe Gln Thr Val Asp Ser Ser Asn Ile Asp Gly Phe Val 340 345 350 Asn Cys Thr Lys Ile Leu Gly Asn Leu Asp Phe Leu Ile Thr Gly Leu 355 360 365 Asn Gly Asp Pro Trp His Lys Ile Pro Ala Leu Asp Pro Glu Lys Leu 370 375 380 Asn Val Phe Arg Thr Val Arg Glu Ile Thr Gly Tyr Leu Asn Ile Gln 385 390 395 400 Ser Trp Pro Pro His Met His Asn Phe Ser Val Phe Ser Asn Leu Thr 405 410 415 Thr Ile Gly Gly Arg Ser Leu Tyr Asn Arg Gly Phe Ser Leu Leu Ile 420 425 430 Met Lys Asn Leu Asn Val Thr Ser Leu Gly Phe Arg Ser Leu Lys Glu 435 440 445 Ile Ser Ala Gly Arg Ile Tyr Ile Ser Ala Asn Arg Gln Leu Cys Tyr 450 455 460 His His Ser Leu Asn Trp Thr Lys Val Leu Arg Gly Pro Thr Glu Glu 465 470 475 480 Arg Leu Asp Ile Lys His Asn Arg Pro Arg Arg Asp Cys Val Ala Glu 485 490 495 Gly Lys Val Cys Asp Pro Leu Cys Ser Ser Gly Gly Cys Trp Gly Pro 500 505 510 Gly Pro Gly Gln Cys Leu Ser Cys Arg Asn Tyr Ser Arg Gly Gly Val 515 520 525 Cys Val Thr His Cys Asn Phe Leu Asn Gly Glu Pro Arg Glu Phe Ala 530 535 540 His Glu Ala Glu Cys Phe Ser Cys His Pro Glu Cys Gln Pro Met Glu 545 550 555 560 Gly Thr Ala Thr Cys Asn Gly Ser Gly Ser Asp Thr Cys Ala Gln Cys 565 570 575 Ala His Phe Arg Asp Gly Pro His Cys Val Ser Ser Cys Pro His Gly 580 585 590 Val Leu Gly Ala Lys Gly Pro Ile Tyr Lys Tyr Pro Asp Val Gln Asn 595 600 605 Glu Cys Arg Pro Cys His Glu Asn Cys Thr Gln Gly Cys Lys Gly Pro 610 615 620 Glu Leu Gln Asp Cys Leu Gly Gln Thr Leu Val Leu Ile Gly Lys Thr 625 630 635 640 His Leu Thr Met Ala Leu Thr Val Ile Ala Gly Leu Val Val Ile Phe 645 650 655 Met Met Leu Gly Gly Thr Phe Leu Tyr Trp Arg Gly Arg Arg Ile Gln 660 665 670 Asn Lys Arg Ala Met Arg Arg Tyr Leu Glu Arg Gly Glu Ser Ile Glu 675 680 685 Pro Leu Asp Pro Ser Glu Lys Ala Asn Lys Val Leu Ala Arg Ile Phe 690 695 700 Lys Glu Thr Glu Leu Arg Lys Leu Lys Val Leu Gly Ser Gly Val Phe 705 710 715 720 Gly Thr Val His Lys Gly Val Trp Ile Pro Glu Gly Glu Ser Ile Lys 725 730 735 Ile Pro Val Cys Ile Lys Val Ile Glu Asp Lys Ser Gly Arg Gln Ser 740 745 750 Phe Gln Ala Val Thr Asp His Met Leu Ala Ile Gly Ser Leu Asp His 755 760 765 Ala His Ile Val Arg Leu Leu Gly Leu Cys Pro Gly Ser Ser Leu Gln 770 775 780 Leu Val Thr Gln Tyr Leu Pro Leu Gly Ser Leu Leu Asp His Val Arg 785 790 795 800 Gln His Arg Gly Ala Leu Gly Pro Gln Leu Leu Leu Asn Trp Gly Val 805 810 815 Gln Ile Ala Lys Gly Met Tyr Tyr Leu Glu Glu His Gly Met Val His 820 825 830 Arg Asn Leu Ala Ala Arg Asn Val Leu Leu Lys Ser Pro Ser Gln Val 835 840 845 Gln Val Ala Asp Phe Gly Val Ala Asp Leu Leu Pro Pro Asp Asp Lys 850 855 860 Gln Leu Leu Tyr Ser Glu Ala Lys Thr Pro Ile Lys Trp Met Ala Leu 865 870 875 880 Glu Ser Ile His Phe Gly Lys Tyr Thr His Gln Ser Asp Val Trp Ser 885 890 895 Tyr Gly Val Thr Val Trp Glu Leu Met Thr Phe Gly Ala Glu Pro Tyr 900 905 910 Ala Gly Leu Arg Leu Ala Glu Val Pro Asp Leu Leu Glu Lys Gly Glu 915 920 925 Arg Leu Ala Gln Pro Gln Ile Cys Thr Ile Asp Val Tyr Met Val Met 930 935 940 Val Lys Cys Trp Met Ile Asp Glu Asn Ile Arg Pro Thr Phe Lys Glu 945 950 955 960 Leu Ala Asn Glu Phe Thr Arg Met Ala Arg Asp Pro Pro Arg Tyr Leu 965 970 975 Val Ile Lys Arg Glu Ser Gly Pro Gly Ile Ala Pro Gly Pro Glu Pro 980 985 990 His Gly Leu Thr Asn Lys Lys Leu Glu Glu Val Glu Leu Glu Pro Glu 995 1000 1005 Leu Asp Leu Asp Leu Asp Leu Glu Ala Glu Glu Asp Asn Leu Ala 1010 1015 1020 Thr Thr Thr Leu Gly Ser Ala Leu Ser Leu Pro Val Gly Thr Leu 1025 1030 1035 Asn Arg Pro Arg Gly Ser Gln Ser Leu Leu Ser Pro Ser Ser Gly 1040 1045 1050 Tyr Met Pro Met Asn Gln Gly Asn Leu Gly Glu Ser Cys Gln Glu 1055 1060 1065 Ser Ala Val Ser Gly Ser Ser Glu Arg Cys Pro Arg Pro Val Ser 1070 1075 1080 Leu His Pro Met Pro Arg Gly Cys Leu Ala Ser Glu Ser Ser Glu 1085 1090 1095 Gly His Val Thr Gly Ser Glu Ala Glu Leu Gln Glu Lys Val Ser 1100 1105 1110 Met Cys Arg Ser Arg Ser Arg Ser Arg Ser Pro Arg Pro Arg Gly 1115 1120 1125 Asp Ser Ala Tyr His Ser Gln Arg His Ser Leu Leu Thr Pro Val 1130 1135 1140 Thr Pro Leu Ser Pro Pro Gly Leu Glu Glu Glu Asp Val Asn Gly 1145 1150 1155 Tyr Val Met Pro Asp Thr His Leu Lys Gly Thr Pro Ser Ser Arg 1160 1165 1170 Glu Gly Thr Leu Ser Ser Val Gly Leu Ser Ser Val Leu Gly Thr 1175 1180 1185 Glu Glu Glu Asp Glu Asp Glu Glu Tyr Glu Tyr Met Asn Arg Arg 1190 1195 1200 Arg Arg His Ser Pro Pro His Pro Pro Arg Pro Ser Ser Leu Glu 1205 1210 1215 Glu Leu Gly Tyr Glu Tyr Met Asp Val Gly Ser Asp Leu Ser Ala 1220 1225 1230 Ser Leu Gly Ser Thr Gln Ser Cys Pro Leu His Pro Val Pro Ile 1235 1240 1245 Met Pro Thr Ala Gly Thr Thr Pro Asp Glu Asp Tyr Glu Tyr Met 1250 1255 1260 Asn Arg Gln Arg Asp Gly Gly Gly Pro Gly Gly Asp Tyr Ala Ala 1265 1270 1275 Met Gly Ala Cys Pro Ala Ser Glu Gln Gly Tyr Glu Glu Met Arg 1280 1285 1290 Ala Phe Gln Gly Pro Gly His Gln Ala Pro His Val His Tyr Ala 1295 1300 1305 Arg Leu Lys Thr Leu Arg Ser Leu Glu Ala Thr Asp Ser Ala Phe 1310 1315 1320 Asp Asn Pro Asp Tyr Trp His Ser Arg Leu Phe Pro Lys Ala Asn 1325 1330 1335 Ala Gln Arg Thr 1340 <210> 116 <211> 345 <212> PRT <213> artificial sequence <220> <223> fusion proteins <400> 116 Glu Glu Glu Leu Gln Val Ile Gln Pro Asp Lys Ser Val Ser Val Ala 1 5 10 15 Ala Gly Glu Ser Ala Ile Leu His Cys Thr Val Thr Ser Leu Ile Pro 20 25 30 Val Gly Pro Ile Gln Trp Phe Arg Gly Ala Gly Pro Ala Arg Glu Leu 35 40 45 Ile Tyr Asn Gln Lys Glu Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Glu Ser Thr Lys Arg Glu Asn Met Asp Phe Ser Ile Ser Ile Ser Asn 65 70 75 80 Ile Thr Pro Ala Asp Ala Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Asp Thr Glu Phe Lys Ser Gly Ala Gly Thr Glu Leu Ser 100 105 110 Val Arg Ala Lys Pro Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro 115 120 125 Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys 130 135 140 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 145 150 155 160 Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 165 170 175 Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 180 185 190 Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His 195 200 205 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 210 215 220 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 225 230 235 240 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu 245 250 255 Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro 260 265 270 Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn 275 280 285 Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 290 295 300 Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val 305 310 315 320 Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln 325 330 335 Lys Ser Leu Ser Leu Ser Pro Gly Lys 340 345 <210> 117 <211> 345 <212> PRT <213> artificial sequence <220> <223> fusion protein <400> 117 Glu Glu Glu Leu Gln Val Ile Gln Pro Asp Lys Ser Val Ser Val Ala 1 5 10 15 Ala Gly Glu Ser Ala Ile Leu His Cys Thr Val Thr Ser Leu Ile Pro 20 25 30 Val Gly Pro Ile Gln Trp Phe Arg Gly Ala Gly Pro Ala Arg Glu Leu 35 40 45 Ile Tyr Asn Gln Lys Glu Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Glu Ser Thr Lys Arg Glu Asn Met Asp Phe Ser Ile Ser Ile Ser Asn 65 70 75 80 Ile Thr Pro Ala Asp Ala Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Asp Thr Glu Phe Lys Ser Gly Ala Gly Thr Glu Leu Ser 100 105 110 Val Arg Ala Lys Pro Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro 115 120 125 Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys 130 135 140 Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val 145 150 155 160 Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 165 170 175 Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 180 185 190 Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His 195 200 205 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 210 215 220 Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 225 230 235 240 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu 245 250 255 Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro 260 265 270 Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn 275 280 285 Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 290 295 300 Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val 305 310 315 320 Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln 325 330 335 Lys Ser Leu Ser Leu Ser Pro Gly Lys 340 345 <210> 118 <211> 347 <212> PRT <213> artificial sequence <220> <223> fusion proteins <400> 118 Glu Glu Glu Leu Gln Val Ile Gln Pro Asp Lys Ser Val Ser Val Ala 1 5 10 15 Ala Gly Glu Ser Ala Ile Leu His Cys Thr Val Thr Ser Leu Ile Pro 20 25 30 Val Gly Pro Ile Gln Trp Phe Arg Gly Ala Gly Pro Ala Arg Glu Leu 35 40 45 Ile Tyr Asn Gln Lys Glu Gly His Phe Pro Arg Val Thr Thr Val Ser 50 55 60 Glu Ser Thr Lys Arg Glu Asn Met Asp Phe Ser Ile Ser Ile Ser Asn 65 70 75 80 Ile Thr Pro Ala Asp Ala Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys 85 90 95 Gly Ser Pro Asp Thr Glu Phe Lys Ser Gly Ala Gly Thr Glu Leu Ser 100 105 110 Val Arg Ala Lys Pro Ser Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro 115 120 125 Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe Pro 130 135 140 Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr 145 150 155 160 Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn 165 170 175 Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg 180 185 190 Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val 195 200 205 Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser 210 215 220 Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys 225 230 235 240 Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu 245 250 255 Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe 260 265 270 Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu 275 280 285 Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe 290 295 300 Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly 305 310 315 320 Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr 325 330 335 Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys 340 345

Claims (31)

Administering to a mammalian subject a therapeutically effective amount of a radionuclide-labeled HER3 targeting agent.
A method of treating solid cancer in a mammalian subject, comprising: The method of claim 1, wherein the radionuclide-labeled HER3 targeting agent is ¹³¹ I, ¹²⁵ I, ¹²³ I, ⁹⁰ Y, ¹⁷⁷ Lu, ¹⁸⁶ Re, ¹⁸⁸ Re, ⁸⁹ Sr, ¹⁵³ Sm, ³² P, ²²⁵ Ac, ²¹³ Bi, ²¹³ Po, ²¹¹ At, ²¹² Bi, ²¹³ Bi, ²²³ Ra, ²²⁷ Th, ¹⁴⁹ Tb, ¹³⁷ Cs, ²¹² Pb or ¹⁰³ Pd, or a combination thereof. 2. The method of claim 1, wherein the radionuclide-labeled HER3 targeting agent comprises a radiolabel selected from ²²⁵ Ac, ¹⁷⁷ Lu, ¹³¹ I, ⁹⁰ Y, ²¹³ Bi, ²¹¹ At, ²¹³ Bi, ²²⁷ Th, ²¹² Pb, or a combination thereof. How to do. The method of claim 1 , wherein the radionuclide-labeled HER3 targeting agent comprises a humanized antibody to HER3. The method of claim 1 , wherein the radionuclide-labeled HER3 targeting agent comprises one or more of patritumab, cerivantumab, rumletuzumab, elgemtumab, AV203, GSK2849330. 2. The method of claim 1, wherein the radionuclide-labeled HER3 targeting agent comprises a monoclonal antibody comprising one or both of a heavy chain sequence comprising SEQ ID NO: 77 and a light chain sequence comprising SEQ ID NO: 78. way of being. The method of claim 1, wherein the radionuclide-labeled HER3 targeting agent is:
(i) (a) an immunoglobulin comprising CDR-H1 comprising SEQ ID NO: 15, CDR-H2 comprising SEQ ID NO: 16, and/or CDR-H3 comprising SEQ ID NO: 17 heavy chain variable region, and
(b) an immunoglobulin light chain variable region comprising CDR-L1 comprising SEQ ID NO: 18, CDR-L2 comprising SEQ ID NO: 19, and/or CDR-L3 comprising SEQ ID NO: 20
either or both;
(ii) one or both of an immunoglobulin heavy chain variable region comprising SEQ ID NO: 21 and an immunoglobulin light chain variable region comprising SEQ ID NO: 22; or
(iii) one or both of the immunoglobulin heavy chain amino acid sequence of SEQ ID NO: 23 and the immunoglobulin light chain amino acid sequence of SEQ ID NO: 24
A method comprising a monoclonal antibody comprising a. The method of claim 1 , wherein the radionuclide-labeled HER3 targeting agent comprises a heavy chain having a complementarity determining region (CDR) having an amino acid sequence as set forth in SEQ ID NO: 13 and/or 1-3, respectively; and/or a light chain having CDRs having the amino acid sequences as set forth in SEQ ID NOs: 14 and/or 4-6, respectively. The method of claim 1, wherein the solid cancer is breast cancer, gastric cancer, bladder cancer, cervical cancer, endometrial cancer, skin cancer, gastric cancer, testicular cancer, esophageal cancer, bronchioloalveolar cancer, prostate cancer, colorectal cancer, ovarian cancer, cervical epidermal cancer, pancreatic cancer, lung cancer, kidney cancer, head and neck cancer, or any combination thereof. The method of claim 1 , wherein the solid cancer is breast cancer, gastric cancer, pancreatic cancer, or any combination thereof. The method of claim 1 , wherein the solid cancer comprises HER3-positive cancer cells. The method of claim 1 , wherein the effective amount of the radionuclide-labeled HER3 targeting agent is the maximum tolerated dose. The method of claim 1, wherein the radionuclide-labeled HER3-targeting agent is ²²⁵ Ac-labeled, and the effective amount of the ²²⁵ Ac-labeled HER3-targeting agent is 0.1 to 50 uCi/kg (body weight of the subject), or 0.1 to 5 uCi/kg (body weight of the subject), or 5 to 20 uCi/kg (body weight of the subject). The method of claim 1, wherein the radionuclide-labeled HER3-targeting agent is ²²⁵ Ac-labeled, and the effective amount of the ²²⁵ Ac-labeled HER3-targeting agent is 2 μCi to 2 mCi, or 2 μCi to 250 μCi, or 75 μCi to 400 μCi A method comprising a dose of The method of claim 1, wherein the effective amount of the radionuclide-labeled HER3 targeting agent is less than 3 mg/kg (subject's body weight), such as 0.001 mg/kg (patient's body weight) to 3.0 mg/kg (patient's body weight), or 0.005 mg/kg (patient's body weight). kg (patient body weight) to 2.0 mg/kg (patient body weight), or 0.01 mg/kg (patient body weight) to 1 mg/kg (patient body weight), or 0.1 mg/kg (patient body weight) to 0.6 mg/kg (patient body weight) body weight), or 0.3 mg/kg (patient body weight), or 0.4 mg/kg (patient body weight), or 0.5 mg/kg (patient body weight), or 0.6 mg/kg (patient body weight) method. The group of claim 1, wherein the radionuclide-labeled HER3-targeting agent consists of once every 7 days, 10 days, 12 days, 14 days, 20 days, 24 days, 28 days, 36 days and 42 days throughout the treatment period. wherein the treatment period comprises at least two administrations. The method of claim 1 , further comprising administering to the subject a therapeutically effective amount of an immune checkpoint therapy, a DNA damage response inhibitor (DDRi), a CD47 blocker, a chemotherapeutic agent, or a combination thereof. 18. The method of claim 17, wherein the immune checkpoint therapy comprises an antibody to PD-1, PD-L1, PD-L2, CTLA-4, CD137, or a combination thereof. 18. The method of claim 17, wherein the DDRi is poly(ADP-ribose) polymerase inhibitor (PARPi), telangiectatic ataxia mutated inhibitor (ATMi), telangiectatic ataxia mutated and Rad-3 related inhibitor (ATRi). ), or a Wee1 inhibitor. 18. The method of claim 17, wherein the CD47 blocker comprises one or more of magnrolimab, remzoparlimab, AO-176, TTI-621, TTI-622, and a modulator of CD47 expression. 18. The method of claim 17, wherein the CD47 blocker comprises a modulator of CD47 expression. 22. The method of claim 21, wherein the modulator of CD47 expression is MBT-001. The method of claim 1 , wherein the radionuclide-labeled HER3 targeting agent is also specific for HER2. 24. The method of claim 1 or 23, wherein the radiolabeled HER3 targeting agent comprises a chemically conjugated chelator group that chelates a radionuclide. 25. The method of claim 24, wherein the chelator group comprises DOTA. 26. The method of any one of claims 1 to 25, wherein the administering step is:
Administering to a subject a therapeutically effective amount of a therapeutic composition comprising a radiolabeled fraction of a HER3 targeting agent and a non-radiolabeled fraction of a HER3 targeting agent.
A method comprising a. 27. The method of claim 26, wherein the therapeutic composition further comprises at least one pharmaceutically acceptable excipient. 28. The method of any one of claims 1-27, further comprising diagnosing the subject having a HER3-positive cancer prior to the administering step. 29. The method of claim 28, wherein the diagnosing step comprises imaging HER3-positive cells in the subject using a radionuclide-labeled HER3 targeting agent. 30. The method of claim 29, wherein the same HER3 targeting agent is used in the diagnosis step and the administration step. 31. The method of claim 30, wherein the HER3 targeting agent is labeled with a different radionuclide in the diagnosis step as in the administration step.

Images