Drugs and treatment methods

Abstract

The present invention relates to an immunoattractant compound (IAC) comprising at least one target structure binding moiety and at least one immune inducer moiety, wherein the target structure binding moiety can bind a target structure to a target cell and / or the target cell environment of a subject, the presence and / or overexpression of the target structure indicates the cancerous disease of the subject, and the immune inducer moiety can induce immune cells to attract immune cells to the target cell and / or the target cell environment.

Description

[Technical Field]

[0001] The present invention relates to an immunoinducing compound (IAC), a pharmaceutical composition comprising an immunoinducing compound (IAC), a method for treating a cancerous disease in a subject, comprising the application of an immunoinducing compound (IAC), a method for inducing an immune response, comprising the step of applying an immunoinducing compound (IAC), a method for determining the effectiveness of at least one treatment, comprising applying an immunoinducing compound (IAC) to a subject, and a method for producing an immunoinducing compound (IAC). [Background technology]

[0002] Cancer is a group of diseases characterized by abnormal cell proliferation that can invade or spread to other parts of the body. Possible signs and symptoms include lumps, unusual bleeding, persistent cough, unexplained weight loss, and changes in bowel movements. More than 100 types of cancer occur in humans, and according to the World Health Organization, cancer is the leading cause of death worldwide, accounting for nearly 10 million deaths in 2020, or roughly one in six deaths. Therefore, the study of effective ways to treat cancer is a focus area of ​​current medical and scientific efforts. While several approaches are currently known to cure, shrink, or halt the progression of cancer, including surgery, radiotherapy or drug therapy, particularly the application of radiotherapy and / or chemotherapy, bone marrow transplantation, immunotherapy, hormone therapy, targeted drug therapy, cryo-thaw necrosis treatment, and radiofrequency ablation, the need for alternative treatment approaches remains high.

[0003] In particular, in cancer immunotherapy, several successful and promising developments have been made recently. These efforts focus on enhancing or inducing a specific immune response in patients to target cancer cells or the tumor immune microenvironment (TIME), and the tumor immune microenvironment (TIME), as shown in more recent studies, is likely to play a crucial role in anti-cancer immunity.

[0004] In particular, tumor-specific antigens (TSAs), which are unique molecules present or that become present on the surface of cancer cells but not present or that do not become present on normal cells, can be used to target cancer cells for destruction by the immune system or other mechanisms. From the immune system's perspective, mutant antigens produced as a result of mutations in the DNA of cancer cells that can lead to the production of abnormal proteins can be recognized as non-self by the immune system and targeted for destruction. Alternatively or in addition to this, overexpressed antigens are present in normal cells but at higher levels in cancer cells and can therefore also be targeted by the immune system or targeted structures. In this regard, TSAs can be used in cancer immunotherapy or radiotherapy. The first is a type of therapy that leverages the immune system's power to fight cancer. For example, vaccines can be developed that target TSAs and thereby stimulate an immune response against cancer cells that express these antigens. The idea of ​​targeting TSAs is hindered by their very close relationship with their non-mutant equivalents (i.e., proteins / peptides with wild-type amino acid sequences) which are recognized as "self" and therefore not recognized and blocked for an effective immune response except for autoimmune antigens. Targeting TSAs presents the problem of triggering off-target reactions, which can result in substantial side effects. The probabilistic nature of mutations leads to antigenic structures that are not optimal for an effective immune response. Therefore, algorithms have been developed to classify mutation-based neoepitopes for vaccination purposes. However, it can be assumed that nearly 70% of tumors, despite having numerous mutations, do not possess effective neoepitopes for vaccination purposes. Furthermore, immunotherapies targeting immune checkpoint molecules, such as checkpoint inhibitor drugs, can be used to release the brakes on the immune system, allowing it to target and destroy cancer cells. However, if tumor recognition is limited due to the lack of optimal neoepitopes or TSAs, releasing the brakes cannot be effective, and this is also true for most tumors, as CPI treatment is effective in only about 20% of tumors for long-term survival.Identifying and characterizing TSA, and more importantly, the ligands that can specifically bind to TSA, is a key area of ​​research in cancer biology, as it has the potential to lead to the development of new and more effective cancer therapies.

[0005] Generally, antigens expressed in cancer cells or the surrounding tumor environment undergo proteasomal degradation after processing in the endoplasmic reticulum (ER), typically into smaller peptides loaded onto MHC class I (MHC-I) molecules. This is primarily driven by "classical" human leukocyte antigen (HLA) genes HLA-A, HLA-B, and HLA-C, which activate immune cell activation, whereas "non-classical" or embryonic HLAs such as HLA-G block immune cell activation, for one reason being secreted into the extracellular stroma as immune cell effectors. The resulting peptide-MHC-I complex is presented on the cell surface for recognition by CD8+ T cells. MHC-II molecules preferentially present peptides originating from exogenous proteins or peptides derived from endogenous proteins that reach the secretory and endocytosis compartments. Binding to MHC-II does not require the same strict sequence and length in the peptide as binding to MHC-I. Therefore, several approaches consider providing mutant peptides or neoantigens that are preferentially presented on MHC-II for immune recognition by CD4+ T cells. Furthermore, tumors have immune cell infiltrations including dendritic cells (DCs), macrophages, and B cells that can act as professional antigen-presenting cells (APCs). Thus, approaches aim to induce an immune response against tumor cells so that the target immune system causes cell death mediated by a targeted immune response. Typically, tumor-infiltrating APCs can activate antigen-specific memory CD4+ and CD8+ T cells that have previously received cognate priming. However, neither cancer cells nor tumor-infiltrating APCs can directly antigen-stimulate T cells. Primary antigen stimulation of naive T cells occurs almost exclusively in lymph nodes (LNs) via highly specialized LN-resident DCs. Simultaneously, research indicates that the tumor immune microenvironment (TIME), which refers to a complex network of interactions between cancer cells, immune cells, and the extracellular matrix within solid tumors, appears to influence the immune system's ability to recognize and attack cancer cells.Factors such as the presence of immunosuppressive cells, the secretion of immunosuppressive cytokines, and the generation of an immunosuppressive extracellular matrix can all contribute to a supportive microenvironment for tumor growth. The immune system is thus suppressed in the tumor immune microenvironment through various mechanisms, including immune checkpoint molecules: tumor cells can express immune checkpoint molecules such as PD-L1, which inhibit the activation of immune cells and prevent them from recognizing and attacking cancer cells; tumor-associated macrophages (TAMs): TAMs are immune cells that can be recruited to tumors and secrete immunosuppressive cytokines such as TGF-β, which suppress the immune response; regulatory T cells (Tregs): Tregs are immune cells that help maintain immune tolerance to autoantigens by suppressing the activity of T cells and other immune cells such as natural killer cells. Treg cells are often abundant in the tumor microenvironment and can contribute to the suppression of the immune response: Myeloid-derived suppressor cells (MDSCs): MDSCs are a population of immune cells that can suppress the immune response through the secretion of immunosuppressive cytokines or HLA fragments and the expression of immune checkpoint molecules; and hypoxia: Hypoxic levels or conditions in the tumor microenvironment can lead to the upregulation of immunosuppressive factors such as HIF-1α and the suppression of the immune response. Furthermore, tumor cells exhibit significant changes in glucose metabolism by switching from the citrate cycle to anaerobic glucose consumption, leading to lactate release, which induces a pH shift to an acidic environment, inhibiting immune cell activity. The immunosuppressive effects of TIME are thought to limit the use of neoantigens.

[0006] An alternative approach to anti-cancer therapy that achieves promising results is offered by theranostics, a term referring to the integration of diagnostic and therapeutic applications in a single approach. Theranostics is a rapidly growing field that aims to optimize patient care by providing diagnosis and treatment simultaneously for specific conditions. The central idea behind theranostics is to develop techniques that can diagnose disease in vivo and deliver therapeutic interventions using the same key molecules for diagnosis against tumors, resulting in a more personalized, efficient, and effective approach to patient care. In this regard, radiotherapy can be combined with molecular imaging techniques such as PET / CT (positron emission tomography / computed tomography), a medical imaging technique that combines two imaging methods, PET and CT, into a single scan.

[0007] Radiation therapy, which uses radionuclides to treat cancer, works by delivering a target dose of ionizing radiation to cancer cells. The radionuclides are usually attached to molecularly targeted agents that specifically bind to tumor cells, allowing the radiation to be delivered to the site of the tumor.

[0008] Radionuclides used in cancer treatment emit either beta particles, which are high-energy electrons or positrons, or alpha particles, which are highly charged particles or Auger electrons. These particles cause damage to the DNA and cell membranes in cancer cells, leading to cell death, particularly in tumors where DNA repair mechanisms are impaired. Radiotherapy with radionuclides can be delivered into the body through parenteral administration of radiolabeled molecules, or as close-range radiotherapy using sealed sources (e.g., iodine-125, iridium-192, rhenium-188, etc.). The advantages of radiotherapy with radionuclides include the ability to deliver high doses of radiation to tumors while minimizing exposure to normal tissue, and the ability to target specific tumor cells, resulting in a more effective and personalized treatment approach. However, the effectiveness of radioligand therapy is also limited, and long-term cures can only be achieved in a small number of cases. As an example, we can cite the recently approved PSMA targeting in patients with metastatic prostate cancer, where "only" 5-10% of patients achieved long-term disease-free survival and 5.3 months of imaging-based progression-free survival compared to standard treatment. 177 Improvement was achieved with Lu-PSMA-617 (Sartor NEJM 2021). Similarly, a dose of 7.4 GB every 8 weeks 177 The response rate for midgut neuroendocrine tumors after four cycles of radioligand therapy with Lu-DOTATATE was 17%, but the progression-free survival rate at 20 months was 65.8% compared to 10.8% in the control group (Strosberg et al NEJM 2017). In summary, while promising, both immunotherapy and radioligand therapy alone have limited efficacy in cancer patients, substantially extending progression-free survival and enabling long-term survival in a very small number of patients with advanced cancer. [Overview of the project]

[0009] Therefore, an object of the present invention is to overcome the above-mentioned problems. In particular, an object of the present invention is to provide compounds suitable for improving cancer treatment. In particular, another object of the present invention is to provide compounds suitable for reducing and / or avoiding the immunosuppressive effects of tumors and / or tumor immune microenvironment (TIME).

[0010] These and other issues are resolved by the subject matter of the attached independent claims.

[0011] The above-mentioned objectives of the present invention are achieved, in particular, by the immunoinducing compounds (IACs) according to the present invention.

[0012] Surprisingly, the inventors have discovered that the use of the immune-attracting compound (IAC) according to the present invention can attract immune cells to tumor sites. Thus, the present invention utilizes the binding of the target structure-binding moiety of the immune-attracting compound (IAC) to the target structure. As described above, the target structure is specific to the tumor site or at least overexpressed, and therefore the compound is likely to preferentially or stochastically bind to cancer cells or structures in the tumor site, particularly to cellular structures in the tumor immune microenvironment (TIME). Thus, such a mechanism advantageously allows for the accumulation of the immune-attracting compound (IAC) in cancer cells or the cancer cell environment. Immune cells are attracted to the tumor site by at least one immune-attracting moiety of the immune-attracting compound (IAC) of the present invention. Thus, the inventors have specifically investigated the possibility of advantageously existing memory responses to the immune system of the target immune system to the immune-attracting moiety, particularly the use of previously known immune-attracting moieties in the target immune system. This is possible through a pre-stimulation or initial antigen stimulation step of the immune system of the subject under treatment, including the application of an immune inducer compound (IAC) according to the present invention, or by the selection of an immune inducer moiety that is or is similar to an antigen to which the immune system of the subject has been previously exposed. For example, and preferably, the immune inducer moiety is an antigen of a vaccine or encodes such an antigen of a vaccine, such as a SARS-CoV-19 vaccine. The inventors believe that antigens to which the subject and its immune system have likely been previously, preferably more than once, exposed in a given population increase the likelihood of immune induction of immune cells to the tumor site. In other words, if a population, such as the population beginning in 2020, has been exposed to a globally epidemic or region-specific viral infection and / or a nearly comprehensive vaccination rate within the population. The application of the immune inducer compound (IAC) of the present invention is particularly advantageous because the previously initially stimulated immune system of the subject to be treated has been previously initially stimulated and is preferably even booster-immunized by the antigen of the immune inducer moiety of the immune inducer compound (IAC) of the present invention.

[0013] The inventors also intended to improve the use of radionuclides that may be included in the immunoinducing compounds (IACs) according to the present invention to convey diagnostic and / or therapeutic effects relating to cancer cells and / or the cancer cell environment. Thereafter, such radionuclides can be selected according to the desired purpose, for example in molecular imaging, to be diagnostic, such as gallium-68 (Ga-68) and fluorine-18 (F-18), or therapeutic, such as lutetium-177 (Lu-177), yttrium-90 (Y-90), and actinium-225 (Ac-225). Such radionuclides can be advantageously included in the concept of immunoinducing compounds (IACs) of the present invention by providing either a chelate for complexing with the radionuclide or a prosthetic group for forming a covalent bond with a nonmetal. In such embodiments in which a radionuclide is included in the immune inducer compound (IAC) according to the present invention, the benefits of theranosticism, i.e., both radionuclide therapy and / or radionuclide diagnostics, can be advantageously combined with the targeted attraction of immune cells to tumor sites using chelates.

[0014] Those with knowledge will immediately recognize that the immune-attracting portion attracts target immune cells to the target site. Thus, the possible damage and / or destruction of target cells by therapeutic radionuclides and / or attracted immune cells, in addition to direct radioactive damage to the cells, can also favorably increase the immune response to the tumor and / or tumor environment through the potent secondary effects of inflammatory stimulation and / or the immune cell-mediated presentation of the respective cell fragments, and therefore, to a certain extent, the respective tumor-associated antigens or tumor-specific antigens.

[0015] Preferred embodiments, including various embodiments as described in the attached claims, can be understood from the dependent claims and otherwise from the following description.

[0016] Therefore, it should be recognized that the present invention is superior to radioligand therapies known in the art, which are largely based on the specific binding of low molecular weight ligands to receptors on the cell surface. The ligand is covalently bound to a chelating agent via a linker structure, so that radionuclides locally deposit various types of radiation onto tumor cells or tumor stroma. The subsequent cytotoxic effect on target cells or target regions is based on damage to DNA, RNA, and proteins. Changes in genetic material result in subsequent novel protein structures, which are recognized as non-self by immune cells, triggering an immune response. Depending on the composition of adult and embryonic MHC complexes and the expression of so-called "checkpoint" factors, this "non-self recognition" leads to a subsequent systemic immune defense response. The immune response can be enhanced, on the one hand, by systemic administration of checkpoint inhibitors (e.g., against PD1 / CTLA4, etc.) directed at immune cells (T cells) that are not normally conjugated, or by administration of immunostimulants. The subsequent immune response is based on the fact that invading immune cells are not stopped by the surface structure of tumor cells.

[0017] In contrast, the concept of "immune induction" using the immune-inducing compounds according to the present invention is based on the principle of recruiting immune cells to tumor cells or sites of prior radioactive ligand enrichment.

[0018] For this purpose, this immunoattractant compound may also advantageously represent an improved theranostic concept.

[0019] Firstly, preferably, typical and well-accepted radioligands, such as SSTRs-, FAP-, CXCR4, or PSMA, can be used as target structure binding sites. During imaging, these target structure binding sites can also be advantageously linked to chelating agents via linkers and subsequent complexes, such as gallium-68 or other isotopes for subsequent imaging. This allows the specificity of enrichment in the tumor region to be visualized and quantified via PET. In the next step, the same radioligand can be used as a target structure binding site to deposit therapeutically effective isotopes near the tumor by chelating them with their therapeutic isotopes (e.g., lutetium-177, yttrium-90, actinium-225, etc.). At least partial cell death is present, and existing immune cells are activated or other immune cells are initially attracted. The latter can be enhanced by covalently binding a synthetic ligand, consisting of ligands for SSTRs-, FAP-, or PSMA, or other targets, to an immune-inducing moiety, which is preferably a highly immunogenic component, such as the coronavirus spike protein or other vaccine agents. Simultaneously or sequentially, with a time delay, the immune response can be triggered by attracting immune cells based on previous vaccination. What is intrinsically present and not originally directed towards the tumor immune response is thus directed towards the tumor or its stroma. This potentially results in an initial moderate local immune response “boost” effect, which can then lead to sustained immunity that prevents tumor recurrence. In one embodiment, the immunogenic component (i.e., the “immune inducer”) may be administered topically to the tumor or tumor stroma.

[0020] With respect to the term “vaccine or vaccination” as used herein, this term is preferably understood to include any form of vaccination, including subcutaneous or other locations, particularly by different vaccination routes, such as oral, intravenous, or spray use.

[0021] The present invention will be described in more detail while referring to the drawings from which further features, embodiments, and advantages can be grasped.

Brief Description of the Drawings

[0022] [Figure 1] FIG. 1 shows a schematic view of an immune-inducing compound according to a first embodiment of the present invention. [Figure 2] FIGS. 2A and 2B show various chemical formulas of chelating molecules. [Figure 3] FIG. 3 shows various schematic and conceptual views of an immune-inducing compound according to the present invention. And [Figure 4] FIGS. 4A and 4B show the results of the first healing test, in particular, the antibody response and the total number of CD eight cells after treatment with an immune-inducing compound according to a second embodiment of the present invention.

Modes for Carrying Out the Invention

[0023] The embodiments in the drawings may relate to preferred embodiments, but all elements and features described in relation to an embodiment can, as appropriate, be combined with any other embodiment and feature discussed herein, particularly in relation to any other embodiment discussed further above. Figure 1 schematically shows an immune inducer compound (IAC) 1 according to a first embodiment of the present invention. Thereafter, the immune inducer compound (IAC) 1 includes a target structure binding moiety 2 that can bind to a target structure 4 of a patient / animal target cell 5 and / or a cell in the target cell environment 6. For understanding, a second cell 51 is shown that is not a target cell and contains a surface molecule 41 that is not bound and / or recognized by the target structure binding moiety 2, in other words, a non-target structure. This difference schematically represents the specific binding of the target structure 4 by the target structure binding moiety 2. If the presence and / or overexpression of the target structure 4 on the target cell 5 indicates the cancerous disease in question, in other words, if the target cell 5 is a cancer cell, it will be understood that the immune inducer compound (IAC) 1 according to the shown embodiment of the present invention selectively binds to the target cell 5 via recognition of the target structure 4 on the target cell 5 and binding to the target structure 4. The immune inducer compound (IAC) 1 according to the shown embodiment also includes an immune inducer moiety 3. In the schematic, simplified example shown, an immune cell receptor 71, such as a B cell receptor, can recognize the immune-inducing portion 3 and therefore can also recognize the binding of immune cells 7 to cancer cells 5 via the immune-inducing compound (IAC) 1. In this schematic example, the immune-inducing portion 3 may be an antigen that has been previously used to vaccinate the subject against a pathogenic infection prior to treatment of the subject with the immune-inducing compound (IAC) 1. For example, the subject may be a cancer patient / animal that has been previously vaccinated with the SARS-CoV-19 vaccine. Thereafter, those skilled in the art will immediately understand that the SARS-CoV-19 vaccine may have provided the antigen not directly as a peptide or protein, but rather in the form of a nucleotide encoding the antigen, such as an mRNA molecule. Thus, each immune-inducing portion 3 may provide the antigen in any form, for example, directly or as part of a peptide, protein or nucleic acid molecule encoding it.In the example shown, for the sake of simplification, the immune-attracting moiety 3 may be an immunodominant epitope of the Sars-CoV-19 spike protein, and subjects may have been inoculated with respective mRNA vaccines whose mRNA encodes the same immunodominant epitope of the Sars-CoV-19 spike protein. In the schematic representation of the principle underlying the present invention, the immune-attracting compound (IAC) 1 is bound to a target structure 4 on a target cell 5, and thus, since the immune-attracting compound (IAC) 1 is bound to a target structure 4 on the cell surface of the target cell 5 in this example, the immune-attracting moiety 3 is also bound to the target cell 5, and immune cells 7 can be attracted to the target cell 5, which is a tumor cell or a cell of the tumor cell environment. Those skilled in the art will immediately understand that the target structure 4 may be the same or different on each tumor cell 5 or each cell of the tumor cell environment 6, but for the sake of brevity, Figure 1 also illustrates the binding of the immune-attracting compound (IAC) 1 to the structural target structure 4 on cells of the tumor immune microenvironment 6. Generally, the same principle as described above applies, and immune cells 7 are attracted to an immune inducer compound (IAC) 1 bound to a target structure 4 of the tumor immune microenvironment 6. Notably, in the schematic representation shown in Figure 1, the immune inducer compound (IAC) 1 includes a linker portion 8 that connects the target structure binding portion 2 and the immune inducer portion 3. Here, the linker portion 8 may contain a chelating agent capable of binding a radioactive portion 9. Several chelating agent molecules are shown in Figures 2A and 2B. These and others may be selected by those skilled in the art with respect to the intended purpose and the specific radioactive portion 9 and immune inducer compound 1 that may be considered for each intended use. In the embodiment shown in Figure 1, the radioactive portion 9 is a radionuclide 9. The radionuclide may be selected according to the desired purpose, for example, to be diagnostic, such as gallium-68 (Ga-68) for molecular imaging, or to be therapeutic, such as lutetium-177 (Lu-177).Since the radionuclides are advantageously incorporated into the concept of the immunoinducing compound (IAC) of the present invention and complexed by chelation, the immunoinducing compound (IAC) 1 can direct the radionuclides to target cells 5 and / or cells in the cellular environment 6. Therefore, the immunoinducing compound (IAC) 1 can be used, for example, to diagnose the presence of target cells 5 in each molecular imaging method, or to deliver a therapeutic effect, i.e., radionuclide therapy by direct irradiation of target cells 5 and / or cells in the tumor cell environment 6, and thus to deliver cancer destruction. Therefore, the present invention, in particular the immune inducer compound (IAC) 1, is suitable for treating target cancerous diseases by synergistically combining a) direct death of tumor cells by irradiation using α- / β emitters, and / or b) attracting immune cells that present, for example, Fc receptors, by binding antibodies to the immune inducer epitope of the IAC, thereby inducing subsequent immune activation by the secretion of immune activators such as IFNG, and / or c) the internal translocation of the IAC compound, which results in the release of peptide fragments to be presented on MHC I or MHC II molecules for interaction in T cell-mediated immune responses, which can then be further enhanced by subsequent checkpoint inhibitor treatment. Fc receptors involved in antibody-mediated recognition of immune inducers include Fcα receptors from the surface of monocytes, macrophages, neutrophils, and eosinophils, as well as Fcγ receptors from the surface of phagocytes, B lymphocytes, NK cells, and dendritic cells, and / or Fcε receptors from the surface of mast cells and basophilic granulocytes. The interaction between the Fc portion of antibodies and Fc receptors (FcRs) on immune cells such as monocytes and macrophages is well known for its immune effector functions, such as the phagocytosis of antibody-coated bacterial and viral particles. Furthermore, Fc-FcR interactions play additional roles related to the regulatory aspects of the immune response. These interactions are involved in antigen presentation, T cell activation and proliferation, and antibody production. As discussed in the background chapter, the application of radioligand therapy, immunotherapy in general, and checkpoint inhibitor approaches alone is fully effective in only a small number of patients. The synergistic nature of these therapeutic approaches is part of the present invention, and this becomes even more effective when both mechanisms are physically connected by immune-inducing compounds that combine target-specific binding and imaging of theranostic compounds with immune activation. In certain preferred embodiments, the target structure binding portion 2 of IAC1 may include affilin 21. The affilin 21 may be advantageously linked to and / or attached to a linker 8, the linker including a half-life extension domain (HEAD) 11. In particular, a chelating agent 91 may also be linked to and / or attached to the linker 8 including the half-life extension domain HEAD 11. In a further embodiment, the immune-inducing portion 3 is linked to and / or attached to a linker 8 containing a half-life extension domain (HEAD) 11. According to the present invention and various embodiments thereof, the immunoinducing compounds (IACs) of the present invention suitable for treating target cancer diseases are: - At least one target structure binding moiety (TSM) 2 and at least one immune attraction moiety (IAM) 3 Includes. Thus, for example, one or more target structure binding moieties and one or more immune-inducing moieties in the form of affilin 21, and any linker 8, if present, in the form of HEAD11 or containing HEAD11; or chelating agent 91 may be arranged in various forms, all of which are considered to be within the scope of the present invention. Several concepts are illustrated in Figure 3. For example, in its simplest form, as shown in Figure 3A, one IAM3 may exist and be linked to one TSM2 by a covalent linker 8. In another example, the IAM1 contains a chelating agent 9. For example, as shown in Figures 3B, 3C, or 3D, respectively, the IAM1 includes two linker portions, a first linker 8a and a second linker 8b. The IAM3 may be linked to a chelating agent 91 that can complexize a radionuclide 9 via the first linker 8a, and the TSM2 may be linked to the chelating agent 91 via the second linker 8b. Alternatively, the chelating agent 91 may be linked to the IAM3 via the first linker 8a, and the TSM2 may be linked to the IAM3 via the second linker 8b. Further alternatively, the chelating agent 91 may be linked to the TSM2 via the first linker 8a, and the IAM3 may be linked to the TSM2 via the second linker 8b. As shown in Figure 3F, the linker structure may also include a first linker connected to a branched linker and / or a second linker, and / or a second linker connected to the first and / or second linker. In the embodiment shown, a three-arm linker is shown connecting IAM3 to TSM2 and chelating agent 91. In a particularly preferred embodiment, the IAC1 according to the present invention includes one or more TSM2 and / or one or more chelating agents 91. As shown in Figure 3F, IAM3 may be connected to TSM2 and / or chelating agent 91 via a first linker 8a, and vice versa. In particular, the IAC1 according to the present invention may include IAM3 connected to TSM2 via a first linker 8a, and the TSM2 connected to chelating agent 91 via a second linker 8b. More specifically, the IAC1 according to the present invention may include a first IAM3 connected to a first TSM2 via a first linker 8a, the TSM2 being connected to a first chelating agent 91 via a second linker 8b, and the IAC1 according to the present invention may include a second IAM3 connected to a second TSM2 via a further linker, the second TSM2 being connected to a second chelating agent 91 via yet another linker. Each of the linkers may be the same as or different from one another. Figures 3G, 3H, 3J, and 3K show several exemplary embodiments of the IAC1, which may include a linker 8 having an affilin 21 and HEAD 11. In particular, Figure 3G illustrates a branched IAC1 in which IAC1 includes an immune-inducing moiety 3 linked to affilin 21 via a first linker, affilin 21 is optionally further linked to a chelating agent 91 via a further linker, and affilin 21 is specifically linked and / or attached to an even further linker having HEAD11. Thus, each linker may independently be the same as or different from any other linker in the molecule. However, in particular, the linker having HEAD11 may be linked to affilin 21. Alternatively, IAC1 may be a linear molecule, as represented by Examples 3H, 3J, and 3K. Here, as shown in Figure 3H, the chelating agent 91 may be optionally present and linked to affilin 21 via a first linker. Affilin 21 may be linked to IAM3 via a second linker, and IAM3 may be linked and / or attached to an even further linker having HEAD11. In a further embodiment, as shown in Figure 3J, IAM3 may be linked via a first linker to an affilin 21 connected and / or attached to a further linker having HEAD11. Furthermore, optionally, a linker 8 having HEAD11 may link the affilin 21 to a chelating agent 91. In further embodiments, as shown in Figure 3K, IAM3 connected to and / or attached to an additional linker having HEAD11 may be linked to affilin 21 via the first linker. Furthermore, optionally, linker 8 having HEAD11 may link IAM3 to a chelating agent 91.

[0024] In the first aspect, the present invention is - At least one target structure binding moiety and at least one immunoattractant moiety Includes, - The target structure binding portion can bind to the target structure of the target cell. - The presence and / or overexpression of the target structure indicates the target cancer disease, - The immune-attracting portion can attract immune cells to the target cells. Regarding immunoassay compounds (IACs).

[0025] The inventors have discovered that the immune-inducing compounds (IACs) according to the present invention remarkably attract immune cells to target cells. Accordingly, the term “target cells” as used herein preferably means cells that are targets of an immune response mediated by the immune-inducing compounds (IACs) of the present invention, such as a cellular immune response. Target cells include, in particular, any undesirable cells, such as cancer cells. Target cells are therefore preferably cells within the body of the subject.

[0026] Therefore, the immune inducer compound (IAC) is selected such that its target structure binding moiety can bind to a target structure known to exhibit the target cancer disease. Such a target structure is selectively expressed from or overexpressed by target cells or cells in the target cell environment, and is thus enriched in the disease area. The immune inducer compound (IAC) according to the present invention is particularly suitable for treating the target cancer disease.

[0027] Accordingly, the present invention utilizes the binding of the target structure-binding portion of an immune-inducing compound (IAC) to the target structure. The target structure is specific to or at least overexpressed in the tumor site of cancer, and therefore the compound is likely to preferentially or stochastically bind to structures in cancer cells or tumor sites, particularly cellular structures in the tumor immune microenvironment (TIME).

[0028] Therefore, such a mechanism advantageously allows for the accumulation of an immune attractant compound (IAC) in cancer cells or the cancer cell environment. Immune cells are attracted to the tumor site by at least one immune attractant moiety of the immune attractant compound (IAC) of the present invention. Therefore, the inventors specifically investigated the possibility that the use of a previously known immune attractant moiety in the target immune system, in particular, may advantageously exist in the memory response of the target immune system to the immune attractant moiety. This is possible in a pre-stimulation or initial antigen stimulation step of the target immune system during therapy, including the application of an immune attractant compound (IAC) according to the present invention, or by the selection of an immune attractant moiety that is or is an antigen to which the target immune system has been previously exposed. For example, and preferably, the immune attractant moiety is an antigen of a vaccine or encodes such an antigen of a vaccine, such as a SARS-CoV-2 vaccine. The inventors hypothesized that antigens to which the target and its immune system have likely been previously, preferably more than once, exposed in a given population increase the likelihood of immune cell attraction to the tumor site. In other words, if a population like the one beginning in 2020 is exposed to a globally prevalent / regionally specific viral infection and / or has a nearly comprehensive vaccination rate within the population, the application of the immune inducer compound (IAC) of the present invention is particularly advantageous because the previously stimulated immune system of the target to be treated has been previously stimulated and is preferably even boosted by the antigen of the immune inducer portion of the immune inducer compound (IAC) of the present invention.

[0029] The inventors also intended to improve the use of radionuclides that may be included in the immunoinducing compounds (IACs) according to the present invention to convey diagnostic and / or therapeutic effects relating to cancer cells and / or cells in the cancer cell environment. Thereafter, such radionuclides can be selected to be diagnostic, for example in molecular imaging, gallium-68 (Ga-68), or therapeutic, lutetium-177 (Lu-177), according to the desired purpose. Such radionuclides can be advantageously incorporated into the concept of the immunoinducing compounds (IACs) of the present invention by providing chelates for providing and complexing the radionuclides. In such embodiments in which radionuclides are incorporated into the immunoinducing compounds (IACs) according to the present invention, the advantages of theranostic, i.e., radionuclide therapy and / or radionuclide diagnostic methods, can be advantageously combined with the targeted attraction of immune cells to tumor sites.

[0030] Those skilled in the art will immediately recognize that, if the immune-inducing portion further includes radionuclides, particularly therapeutic radionuclides, the target immune cells may be favorably attracted to the target site. Thereafter, the possible damage and / or destruction of target cells by the therapeutic radionuclides and / or attracted immune cells may also favorably increase the immune response to the tumor and / or tumor environment through inflammatory stimuli and / or the potent secondary effect of the immune cell-mediated presentation of the respective cell fragments, and therefore, with a certain possibility, the respective tumor-associated antigens or tumor-specific antigens.

[0031] Preferred embodiments, including various embodiments as described in the attached claims, can be understood from the dependent claims and otherwise from the following description.

[0032] Although the present invention is described in detail below, please understand that the methods, protocols, and reagents may vary, and therefore the present invention is not limited to the specific methods, protocols, and reagents described herein. Please also understand that the terms used herein are intended solely to describe specific embodiments and are not intended to limit the scope of the present invention, which is limited only by the appended claims. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art.

[0033] Preferably, the terms used herein are defined as those described in “A multilingual glossary of biotechnological terms: (IUPAC Recommendations)”, H.G. W. Heuenberger, B. Nagel, and H. Kolbl, Eds., (1995) Helvetica Chimica Acta, CH-4010 Basel, Switzerland.

[0034] The implementation of this invention will, unless otherwise specified, utilize conventional methods of biochemistry, cell biology, immunology, and recombinant DNA techniques as described in the literature in the art (e.g., Molecular Cloning: A Laboratory Manual, 2nd Edition, J. Sambrook et al. eds., Cold Spring Harbor Laboratory Press, Cold Spring Harbor 1989).

[0035] As used herein, the term “target structure binding moiety” preferably refers to any component of a molecule or compound that specifically binds to the target structure, such as a receptor on a cell surface. More preferably, the target structure binding moiety is designed to selectively bind to a particular target structure, enabling the delivery of the immune inducer compound to a target site, such as a target cell or the target cell environment. Specific binding between the target structure binding moiety and the target structure is important in ensuring the effective delivery of the immune inducer compound (IAC) to the target cell and / or the target cell environment.

[0036] Throughout this specification and the subsequent claims, unless the context requires otherwise, the word “comprise,” and variations such as “comprises” and “comprising,” are understood to mean that they encompass the described component, integer or process or group of components, integers or processes, but not any other component, integer or process or group of components, integers or processes; however, in some embodiments, such other components, integers or processes or groups of components, integers or processes may be excluded, i.e., the subject matter lies in the inclusion of the described component, integer or process or group of components, integers or processes. The terms “a,” “an,” and “the” and similar references used in the context describing the invention (particularly in the context of the claims) should be interpreted as encompassing both singular and plural, unless otherwise indicated herein or unless clearly inconsistent with the context. The descriptions of ranges of values ​​herein are intended merely as a concise way of referring individually to each separate value that falls within that range. Unless otherwise indicated herein, each individual value is incorporated herein as if it were listed separately herein.

[0037] All methods described herein may be performed in any suitable order, unless otherwise indicated herein or unless it is clearly inconsistent with the context. Any use of any examples or illustrative language provided herein (e.g., "etc.") is intended merely to better illustrate the invention and not to impose any limitation on the scope of the invention as claimed elsewhere. No language herein should be construed as indicating an unclaimed element essential to the practice of the invention.

[0038] As used herein, the term “immune attractant moiety” preferably refers to a molecule or group of molecules that can attract immune cells to a specific site in the body, target cells and / or cells adjacent to target cells, i.e., the target cell environment. In the context of the present invention, the effect of including an immune attractant moiety in the immune attractant compound (IAC) of the present invention is to attract immune cells to target cells and / or cells in the target cell environment, which can then transmit an effective immune response to the target cells and / or cells in the target cell environment, and thus can increase the effectiveness of the immune response against tumors.

[0039] In a further preferred embodiment, the immune inducer compound (IAC) according to the present invention can therefore induce an immune response, particularly a cellular immune response, to the immune inducer moiety.

[0040] In a further preferred embodiment, the immune inducer compound (IAC) according to the present invention can therefore induce an immune response, particularly a cellular immune response, against the immune inducer moiety or the corresponding cells in the target.

[0041] In a further preferred embodiment, the immune-inducing compound (IAC) according to the present invention can therefore leverage the existing immune cell recognition of an epitope to increase the immune cell volume and / or cell death activity of cells in target cells and / or the target cell environment.

[0042] As used herein, the term “target cell environment” preferably refers to the microenvironment in which target cells are located, preferably called the tumor stroma, particularly in the context of tumors. Thus, the underlying idea of ​​the inventors is that the target cell environment exhibits a concentration of immune cells, and these immune cells can mediate an immune response. This encompasses the possibility of initiating both specific and nonspecific immune responses. Specifically, a nonspecific immune response may involve the infiltration of immune cells into the target cell environment that do not specifically target the intended cells. In the case of tumors, the tumor stroma may encompass a variety of cells and extracellular components, including fibroblast-like cells, endothelial cells, and immunomodulatory cells. The concentration of immune cells in the target cell environment, particularly within the tumor stroma, can facilitate an efficient immune response against target cells containing tumor-associated antigens, thereby helping to enhance the effectiveness of immunotherapeutic approaches.

[0043] The term "immune response" refers to the integrated bodily response to a target such as an antigen, preferably a cellular immune response or a cellular and humoral immune response. Immune responses can be protective / preventive / protective and / or therapeutic.

[0044] "To induce or trigger an immune response" may mean that no immune response existed before induction, but it may also mean that a certain level of immune response existed before induction and that the immune response is enhanced after induction. Therefore, "to induce an immune response" also includes "to enhance an immune response." Preferably, after inducing an immune response in a subject, the subject is protected from developing a disease such as cancer, or the disease state is improved by inducing the immune response. For example, an immune response to tumor-expressing antigens may be induced in patients / animals with cancer or in subjects at risk of developing cancer. In this case, inducing an immune response may mean that the disease state of the subject is improved, that the subject does not metastasize, or that a subject at risk of developing cancer does not develop cancer.

[0045] The terms “cellular immune response” and “cellular response,” or similar terms, refer to an immune response directed at cells characterized by the presentation of antigens by class I or class II MHC accompanied by T cells or T lymphocytes acting as either “helper” or “killer.” Helper T cells (also called CD4+ T cells) play a central role by regulating the immune response, while killer cells (also called cytotoxic T cells, cytolytic T cells, CD8+ T cells, or CTLs) kill disease cells such as cancer cells and prevent the production of more disease cells. In preferred embodiments, the present invention includes stimulating an antitumor CTL response against tumor cells that express one or more tumor expression antigens, preferably presenting such tumor expression antigens together with class I MHC.

[0046] Such immune responses are advantageous in eliminating target cells and / or cells in the target cell environment, or at least in preventing further cell division of tumor cells and thus tumor growth. Another favorable effect of attracting immune cells to target cells and / or cells in the target cell environment may further include any new immunologically active substances released by tumor cell apoptosis encountering an "inflammatory environment" in which the immune response can be amplified. The concept of IACs may also be equally applicable to the angiogenesis process, provided that appropriate target structures for binding are identified. Furthermore, it may favorably alter angiogenesis and blood and substrate supply.

[0047] Therefore, it is important to understand that an immune response induced against an immune inducer may include a series of events occurring in the target cells or target cell environment of the subject, particularly when the target cells or target cell environment of the subject are exposed to the immune inducer. The response may involve the activation of various cells and molecules of the immune system, such as antibodies, T cells, and cytokines. Thus, the immune response is naturally directed towards the immune inducer, ultimately eliminating or neutralizing it, and therefore eliminating or neutralizing the target cells and / or target cell environment. In particular, immune responses in morphology or T cell responses may be advantageous in bringing about the desired therapeutic effect of immune inducer compounds in the treatment of cancer. It is known to those skilled in the art that T cell-mediated cellular immune responses are an important component of the adaptive immune system that helps protect the body from infections and abnormal cells. This process begins when T cells, also known as T lymphocytes, encounter a foreign antigen presented on the surface of an antigen-presenting cell (APC). "Antigen-presenting cells" (APCs) are cells that present peptide fragments of protein antigens associated with MHC molecules on their cell surface. Some APCs can activate antigen-specific T cells. Professional antigen-presenting cells are highly efficient at taking in antigens either through phagocytosis or receptor-mediated endocytosis, and then presenting antigen fragments bound to class II MHC molecules on their membranes. T cells recognize and interact with the antigen-class II MHC molecule complex on the membrane of the antigen-presenting cell. Subsequently, additional co-stimulatory signals are produced by the antigen-presenting cell, leading to T cell activation. The expression of co-stimulatory molecules is a defining characteristic of professional antigen-presenting cells.

[0048] In the innovative concept of the present invention, it is particularly advantageous if an immune inducement moiety containing an antigen known to the target immune system is selected, especially if it is due to a previous vaccination or pathogen infection, such as SARS-COVID-19 infection and / or vaccination. This is because APCs, such as dendritic cells, macrophages, and B cells, as antigen-presenting cells, take up and process antigens and form complexes with major histocompatibility complex (MHC) molecules to present fragments of the antigen on their cell surface. T cells expressing T cell receptors (TCRs) recognize the foreign antigen presented on the MHC molecules and bind to the APCs. When T cells encounter the antigen presented on the APCs, they undergo activation and proliferation processes, resulting in the production of many identical T cells that are specific to the same antigen. These activated T cells can then directly attack infected or abnormal cells by releasing cytokines, or they can differentiate into effector T cells, such as cytotoxic T cells or helper T cells, which have specialized functions in the immune response.

[0049] Cytotoxic T cells, also known as CD8+ T cells, can directly kill infected or abnormal cells through a process called cytotoxicity. Helper T cells, also known as CD4+ T cells, play a crucial role in integrating the immune response by producing cytokines that activate and regulate the activity of other immune cells, including B cells and cytotoxic T cells. Thus, the T cell response is an essential aspect of the adaptive immune system that enables the body to respond rapidly and effectively to new or altered infections and abnormal cells. In the context of this invention, the target structure is bound by a target structure binding moiety of an immune inducer compound (IAC), and therefore the T cell induces its effect on the target cell and / or target cell environment mediated by the T cell's recognition of the immune inducer moiety. This response is therefore considered to be particularly related to inducing a targeted immune response of T cells directed against the target cell and / or target cell environment, preferably. In this regard, prior contact with each immune-inducing moiety, particularly in the form of vaccination by the respective immune-inducing moiety or pathogen infection by the immune-inducing moiety, is particularly advantageous and is thought to enhance the therapeutic effect of the immune-inducing compound of the present invention on the cancer the subject is suffering from. This is because, as with such prior infection and / or vaccination, the subject's immune system is already stimulated to respond more effectively and / or more rapidly to the immune-inducing moiety. In particular, a memory T cell and / or memory B cell response may be triggered. Memory T cells are a type of T cell that plays a crucial role in the immune response to recurrent infections. This phenomenon is utilized in the present invention, where the immune response is redirected to the target cell and / or target cell environment by the presentation of an immune-inducing moiety contained in the immune-inducing compound (IAC), preferably bound to the target cell and / or target cell environment via a target structure binding moiety. Memory T cells are produced after a person is first exposed to a pathogen, either through vaccination with the pathogen's antigen or infection by the pathogen itself. If the pathogen infects the body again in the future, these memory T cells can recognize and respond to the same pathogen.The memory T cell response process is thought to involve an initial antigen-stimulating event at the first point of exposure to a pathogen, where T cells (including CD4+ and CD8+ T cells) are activated and differentiate into effector T cells that can recognize and respond to specific antigens on the pathogen. Expansion follows initial antigen stimulation: after initial activation, effector T cells proliferate and increase in number to provide a stronger immune response. Memory T cells are then formed: some of the effector T cells then differentiate into memory T cells, which have a longer lifespan than effector T cells and can remain in the body for longer periods. Using the immune-inducing compound of the present invention, the application of the compound mimics a recurrent infection in which memory T cells rapidly recognize and respond to the immune-inducing portion when it is considered a "pathogen," providing a rapid and strong immune response against target cells. This is because memory T cells have already been antigen-stimulated to specifically recognize the immune-inducing portion, allowing them to respond more rapidly and effectively compared to naive T cells. Accordingly, depending on the specific composition and construction of the immune-inducing compound (IAC) of the present invention, its application may also include an enhancing effect in which, upon re-exposure of the subject to the immune-inducing site, memory T cells are reactivated and their numbers increase, further enhancing the immune response against the immune-inducing site. This process can lead to the development of more memory T cells, which can provide long-term protection against future infections. Overall, the memory T cell response is a crucial component of the immune system that helps protect against recurrent infections by providing a rapid and robust immune response, and in the context of the present invention, it is considered to induce an effective immune response directed against the immune-inducing site.

[0050] The main types of professional antigen-presenting cells are dendritic cells, macrophages, B cells, and certain activated epithelial cells, which possess the broadest range of antigen presentation and are perhaps the most important antigen-presenting cells. Dendritic cells (DCs) are a population of leukocytes that present antigens captured in peripheral tissues to T cells via both MHC class II and I antigen presentation pathways. It is well known that dendritic cells are potent inducers of the immune response, and that the activation of these cells is a crucial step for inducing antitumor immunity. Dendritic cells are conveniently classified as “immature” and “mature” cells, which can be used as a simple way to distinguish between two well-characterized phenotypes. However, this nomenclature should not be interpreted as excluding all possible intermediate stages of differentiation. Immature dendritic cells are characterized as antigen-presenting cells with a high capacity for antigen uptake and processing, and this high capacity correlates with high expression of Fcγ receptors and mannose receptors. The mature phenotype is typically characterized by lower expression of these markers, but high expression of cell surface molecules responsible for T cell activation, such as class I and class II MHCs, adhesion molecules (e.g., CD54 and CD11), and co-stimulatory molecules (e.g., CD40, CD80, CD86, and 4-1BB). Dendritic cell maturation is called the dendritic cell activation state in which such antigen-presenting dendritic cells result in initial antigen stimulation of T cells, while presentation by immature dendritic cells results in tolerance. Dendritic cell maturation is primarily triggered by biomolecules with microbial characteristics detected by innate receptors (bacterial DNA, viral RNA, endotoxins, etc.), pro-inflammatory cytokines (TNF, IL-1, IFN), ligation of CD40 on the dendritic cell surface by CD40L, and substances released from cells undergoing stress-induced cell death. Dendritic cells can be induced in vitro by culturing myeloid cells with cytokines such as granulocyte-macrophage colony-stimulating factor (GM-CSF) and tumor necrosis factor α.

[0051] Nonprofessional antigen-presenting cells do not constitutively express MHC class II proteins required for interaction with naive T cells; these are expressed only when stimulated by certain cytokines, such as IFNγ. Antigen-presenting cells can be loaded with MHC class I-presented peptides by transducing cells with nucleic acids, preferably RNA, that encode peptides or polypeptides containing the peptides to be presented, such as nucleic acids encoding antigens or polypeptides used for vaccination.

[0052] According to the present invention, the term "antigen-presenting cell" preferably also includes target cells.

[0053] Additionally or alternatively, the immune-inducing compounds (IACs) according to the present invention may be effective in treating cancer by inducing a B-cell response, also known as a humoral immune response, directed towards the immune-inducing moiety. The B-cell response is known to be a more important key part of the adaptive immune system, inherently responsible for recognizing and neutralizing foreign pathogens in the target body. Thus, B cells possess surface receptors called antibodies that are specific to certain antigens (molecules or substances that are non-self to the body). When a B cell encounters an antigen that fits its antibody receptor, the B cell binds to that antigen. The binding of the antigen to the antibody receptor on the B cell triggers a series of molecular and cellular events that lead to B cell activation. This activation process involves signaling pathways that result in the production of new antibody molecules and the expansion and differentiation of B cells into plasma cells. Activated B cells differentiate into plasma cells, which are specialized cells that produce and secrete large amounts of antibody molecules into the bloodstream. Antibodies produced by plasma cells are specific to the antigen that triggered the B-cell response and can bind to and neutralize the antigen. Antibodies produced by plasma cells circulate in the bloodstream and can bind to antigens present in the body. This binding can neutralize the antigen by blocking its function or by marking it for destruction by other cells of the immune system, such as phagocytic cells. Therefore, in the context of this invention, those skilled in the art will immediately recognize that an antibody response directed against the immune-inducing moiety of an immune-inducing compound (IAC) can thus result in an effective immune response against target cells and / or target cell structures exhibiting the target structure to which the immune-inducing compound (IAC) binds via its target structure-binding moiety. Similar to memory T cell responses, some activated B cells that survive the initial response also differentiate into memory B cells, which remain in the body and can respond more rapidly and effectively to subsequent exposure to the same antigen. This is why vaccination, which exposes the immune system to a harmless form of a pathogen, can provide long-lasting immunity against future infections by the same pathogen.Therefore, in particular, if the target immune system has previously received initial stimulation at the immune-inducing portion due to a previous infection and / or vaccination, the memory B cell response is also considered effective in therapies using the immune-inducing compounds of the present invention.

[0054] In a further preferred embodiment, the immune-inducing portion includes an antigen, preferably the antigen is known in advance to the target immune system.

[0055] As used herein, the term “antigen” preferably refers to any substance that is a target of an immune response, such as a specific reaction with an antibody or a T lymphocyte (T cell), and / or induces an immune response, such as a specific reaction with an antibody or a T lymphocyte (T cell), preferably a substance comprising a protein. Preferably, the antigen comprises at least one epitope, such as a T cell epitope. Preferably, the T cell epitope, when presented by MHC and recognized by a T cell receptor, can induce clonal proliferation of T cells having a T cell receptor that specifically recognizes the peptide / MHC complex in the presence of appropriate co-stimulatory signals. Preferably, the T cell epitope comprises an amino acid sequence substantially corresponding to the amino acid sequence of a fragment of the antigen. Preferably, the fragment of the antigen is a peptide presented by MHC class I and / or class II. The T cell epitopes according to the present invention preferably relate to a part or fragment of an antigen that can stimulate a cellular response to cells, such as disease cells, particularly cancer cells, characterized by an immune response, preferably by the expression of or to the antigen, and preferably by the presentation of the antigen. Preferably, the T cell epitope can stimulate a cellular response to cells characterized by antigen presentation by class I MHC, and more preferably, it can stimulate antigen-responsive cytotoxic T lymphocytes (CTLs).

[0056] Preferably, in the context of the present invention, an antigen is optionally a molecule that, after processing, preferably induces a specific immune response against the antigen (including cells expressing the antigen). The antigen or its T cell epitope is presented by cells, preferably in the context of MHC molecules, by antigen-presenting cells, including disease cells, particularly cancer cells, to elicit an immune response against the antigen (including cells expressing the antigen).

[0057] The term "Major Histocompatibility Complex" and the abbreviation "MHC" refer to a complex of genes that occur in all vertebrates, encompassing MHC class I and MHC class II molecules. MHC proteins or molecules are important for signaling between lymphocytes and antigen-presenting or disease cells in immune responses. MHC proteins or molecules bind peptides and present them for recognition by T cell receptors. Proteins encoded by MHC are expressed on the cell surface and present both self-antigens (peptide fragments from the cell itself) and non-self-antigens (e.g., fragments of invading microorganisms) to T cells. The MHC region is divided into three subgroups: class I, class II, and class III. MHC class I proteins contain the α chain and β2 microglobulin (encoded by chromosome 15 and not part of MHC). MHC class I proteins present antigen fragments to cytotoxic T cells. On most immune system cells, and especially on antigen-presenting cells, MHC class II proteins contain the α and β chains and present antigen fragments to T helper cells. The MHC class III region encodes complement components and other immune components, such as some cytokines. The MHC is polygenic (several MHC class I and MHC class II genes exist) and polymorphic (multiple alleles exist for each gene). As used herein, the term “haplotype” refers to an HLA allele found on a single chromosome and the protein it encodes. A haplotype may also refer to an allele present at any single locus within the MHC. Each class of MHC is represented by several loci: for example, for class I, HLA-A (human leukocyte antigen-A), HLA-B, HLA-C, HLA-E, HLA-F, HLA-G, HLAH, HLA-J, HLA-K, HLA-L, HLA-P, and HLA-V; and for class II, HLA-DRA, HLA-DRB1~9, HLA-, HLA-DQA1, HLA-DQB1, HLA-DPA1, HLA-DPB1, HLA-DMA, HLA-DMB, HLA-DOA, and HLA-DOB. The terms "HLA allele" and "MHC allele" are used interchangeably herein.MHC exhibits extreme polymorphism, and within the human population, there are numerous haplotypes at each locus containing different alleles. Different polymorphic MHC alleles, both class I and class II, have different peptide specificities, and each allele encodes a protein that binds peptides exhibiting a specific sequence pattern. In one preferred embodiment of all aspects of the present invention, the MHC molecule is an HLA molecule. According to the present invention, MHC class II includes HLA-DM, HLA-DO, HLA-DP, HLA-DQ, and HLA-DR. In the context of the present invention, the term “MHC-binding peptide” includes MHC class I and / or class II binding peptides, or peptides that can be processed to produce MHC class I and / or class II binding peptides. In the case of class I MHC / peptide complexes, the binding peptide is typically 8–12 amino acid long, preferably 8–10 amino acid long, but longer or shorter peptides may be effective. In the case of class II MHC / peptide complexes, the bound peptide is typically 9–30 amino acids long, preferably 10–25 amino acids long, and particularly 13–18 amino acids long, although longer and shorter peptides may be effective. When the peptide is presented directly, i.e., without processing, and especially without cleavage, the peptide has a length suitable for binding to MHC molecules, particularly class I MHC molecules, and is preferably 7–30 amino acids long, such as 7–20 amino acids long, more preferably 7–12 amino acids long, more preferably 8–11 amino acids long, and particularly 9 or 10 amino acids long. When the peptide is part of a larger entity containing further sequences, such as a vaccine sequence or polypeptide, and is presented after processing, especially after cleavage, the peptide produced by processing has a length suitable for binding to MHC molecules, particularly class I MHC molecules, and is preferably 7–30 amino acids long, such as 7–20 amino acids long, more preferably 7–12 amino acids long, more preferably 8–11 amino acids long, and particularly 9 or 10 amino acids long.Preferably, the peptide sequence presented after processing is derived from the amino acid sequence of the antigen or polypeptide used for vaccination; that is, the sequence substantially corresponds to, and preferably is completely identical to, the fragment of the antigen or polypeptide. Therefore, the MHC-binding peptide in one embodiment contains a sequence that substantially corresponds to, and preferably is completely identical to, the fragment of the antigen.

[0058] In preferred embodiments of the immunoinducing compound (IAC) according to the present invention, the immunoinducing portion includes an antigen expressed by target tumor cells or cells in the tumor cell environment.

[0059] Those skilled in the art will immediately recognize and acknowledge that, in this context, the term “tumor antigen” may refer to a specific antigenic substance, preferably originating from tumor cells or the tumor cell environment within the subject. It is extremely important to note that, in this application, “antigen” is preferably used in a dual sense. Firstly, an antigen may function as a “target,” representing a specific molecular entity against which an immune response is desired. In addition, or simultaneously, the term “antigen” may be used as an “immune attractant,” meaning a component that can attract and activate immune cells within the target cell environment. Thus, those skilled in the art will be able to understand each phrase in context and distinguish between the two aforementioned contexts, preferably, the term “antigen” is used in relation to a specific target of an immune response, while the term “antigen” preferably acts as an immune attractant, promoting the concentration and activation of immune cells in the desired environment.

[0060] More preferably, the antigen is a tumor-specific antigen (TSA). The term “tumor-specific antigen (TSA)” is therefore understood, preferably in the context of the present invention, as a molecule or substance that is specifically expressed in tumor cells but not in normal, healthy cells. TSAs can be particularly thought to be proteins or peptides expressed in tumor cells that may originate from the cytoplasm, cell surface, or cell nucleus, and which are part of tumor cells, particularly those that exist primarily intracellularly or as surface antigens of tumor cells. For example, tumor antigens include carcinoembryonic antigens, α1-fetoprotein, isoferritin and fetal sulfoglycoprotein, α2-H-ferroprotein and γ-fetoprotein. According to the present invention, tumor antigens preferably include any antigen expressed in tumors or cancers and tumor or cancer cells, and optionally any antigen that is characteristic in terms of type and / or expression level for tumors or cancers and tumor or cancer cells, i.e., tumor-associated antigens. In one embodiment, the term “tumor-associated antigen” refers to a protein that is specifically expressed under normal conditions in a limited number of tissues and / or organs or at a particular developmental stage, for example, a tumor-associated antigen may be specifically expressed under normal conditions in gastric tissue, preferably in the gastric mucosa, in reproductive organs, for example in the testes, in chorionic tissue, for example in the placenta, or in germline cells, and may be expressed or abnormally expressed in one or more tumor or cancerous tissues. In this context, “limited number” means preferably three or less, more preferably two or less. Tumor antigens in the context of the present invention include, for example, differentiation antigens, preferably cell type-specific differentiation antigens, i.e., proteins that are specifically expressed under normal conditions in a particular cell type at a particular differentiation stage, cancer / testicular antigens, i.e., proteins that are specifically expressed in the testes and sometimes in the placenta under normal conditions, and germline-specific antigens. Preferably, tumor antigens or abnormal expression of tumor antigens identify cancer cells. In the context of the present invention, tumor antigens expressed by cancer cells in a subject, e.g., a patient / animal suffering from cancerous disease, are preferably autoproteins in the subject.In preferred embodiments, tumor antigens in the context of the present invention are specifically expressed under normal conditions in non-essential tissues or organs, i.e., tissues or organs that do not result in the death of the subject when damaged by the immune system, or in organs or structures of the body that are inaccessible or barely accessible by the immune system.

[0061] These antigens are recognized as non-self by the immune system and can be targeted by immune cells to attack and destroy tumor cells. Therefore, using TSA as the immune-inducing moiety of the immune-inducing compound of the present invention can enhance the immune response against target cells and / or the target cell environment. Tumor-specific antigens can originate from a variety of sources, including mutant proteins specific to tumor cells and abnormal cell surface molecules that are expressed differently on tumor cells compared to normal healthy cells. Tumor-specific antigens are of great interest to the development of cancer treatments because they can be used as targets for immune-based therapies such as vaccines and immunotherapies. These therapies aim to train the immune system to specifically recognize and destroy tumor cells without affecting normal healthy cells. The concept of the present invention can utilize this knowledge and thus result in the specific and targeted death or destruction of tumor cells and / or the tumor cell environment. Thus, it is considered that those skilled in the art are familiar with a variety of tumor-specific antigens and can select tumor-specific antigens as possible immune-inducing moieties of the immune-inducing compound of the present invention for the desired purposes of the present invention. In particular, tumor-specific antigens may be antigens specific to the cancerous disease in question. This can advantageously achieve amplification of the immune response against target cells and / or the target cell environment.

[0062] In a further preferred embodiment, the tumor-specific antigen may be selected to be an antigen specific to a cancer disease different from the cancer disease of interest. This may be particularly advantageous if a TSA known to induce a potent and specific immune response is selected. The use of a TSA is particularly preferred because it is thought not to be present on the healthy cells of interest. It is important to note that not all tumors express unique or recognizable antigens, and some tumors may express antigens that are also present on normal healthy cells. Therefore, the identification and validation of tumor-specific antigens is an ongoing area of ​​research in the fields of cancer biology and immunotherapy, which is carefully followed by those skilled in the art. Alternatively or additionally, the immune-inducing portion may include an antigen not expressed by the tumor cells or tumor cell environment of interest.

[0063] In a further preferred embodiment, the immune-inducing portion includes an antigen that is not expressed by the target tumor cells or cells in the tumor cell environment.

[0064] Since antigens can be selected independently of tumors, this can be particularly advantageous. In particular, since factors in tumors, especially in the tumor environment TIME, are known to inhibit immune responses, it can be advantageous to select antigens and / or antigens of a specific affinity that are known to induce a particular strong immune response. More importantly, since vaccination with the following antigens and / or infection with such antigens can be assumed with a high probability, the antigens may be antigens that have been previously exposed to the target immune response, or at least antigens that are likely to have been previously exposed to the target immune response. However, artificial non-pathogenic antigens can also be considered as immune-inducing moieties in the immune-inducing compounds of the present invention.

[0065] In a further preferred embodiment, the immune-inducing portion includes an antigen selected from pathogen-derived antigens, tumor immune microenvironment (TIME)-related antigens, tumor-associated antigens (TAAs), tumor-specific antigens (TSAs), tumor germline antigens, neoantigens, artificial immunostimulatory antigens, or combinations thereof.

[0066] As used herein, the term “pathogen-derived” preferably refers to something that originates from or is derived from a pathogen. Thereafter, a pathogen is understood to be a microorganism, such as a bacterium, virus, fungus, or parasite, that can cause disease or infection in its host organism. In other words, where an entity or substance is described as “pathogen-derived,” it preferably means that the entity or substance comes from or is associated with a pathogen. For example, in relation to antigens, therapies, or compounds, a “pathogen-derived antigen” is preferably an antigen that originates from or is associated with a pathogen. Such antigens are known to those skilled in the art as being used in a variety of applications, including the development of vaccines or immunotherapies aimed at inducing an immune response against a particular pathogen or cells infected with a pathogen.

[0067] According to the present invention, the term “derived” as used herein preferably means that a particular entity, in particular a particular peptide sequence, is present in the object from which it originates. In the case of an amino acid sequence, in particular a particular sequence region, “derived” means, in particular, that the amino acid sequence originates from an amino acid sequence in which the amino acid sequence exists.

[0068] According to the present invention, the term "neoantigen" refers to a peptide or protein that includes one or more amino acid modifications compared to the parent peptide or protein. For example, a neoantigen may be a tumor-associated neoantigen, and the term "tumor-associated neoantigen" encompasses peptides or proteins that include amino acid modifications due to tumor-specific mutations. In this regard, it is important to understand that the immune response to tumor antigens, particularly mutated tumor antigens, is not induced by the tumor cells themselves, but rather by antigen-presenting cells, especially dendritic cells, that receive tumor antigens released from tumor cells. It is also known that, in order to achieve an effective immune response, the released tumor antigens taken up by antigen-presenting cells must be processed and presented by MHC class II (exogenous presentation) for the induction of a CD4 immune response, or by MHC class I (cross-presentation) for the induction of a CD8 immune response. For the latter immune response, the presence of a CD4 immune response to the same or different tumor antigens delivered to the same antigen-presenting cells is necessary (Bennett et al., J.Exp.Med.186,65-70(1997)). It is known in the art that the cellular localization of antigens in disease cells such as tumor cells can determine whether those antigens are taken up and presented by antigen-presenting cells. Exosomes released from disease cells such as tumor cells contain mRNA, proteins, and MHC peptide complexes, and therefore can transmit these components to antigen-presenting cells. Exosomes are produced by invagination and therefore mainly contain cytoplasmic components in addition to endocytosis membrane molecules. Therefore, it is thought that cytoplasmic components such as proteins are concentrated in exosomes and can be transmitted to antigen-presenting cells. Exosomes can also productively transmit mRNA, which can be translated in cells that take up RNA.Therefore, although we do not wish to be bound by any particular theory, since exosomes are taken up by antigen-presenting cells, and peptides and proteins are presented by antigen-presenting cells (optionally, after translation of coding RNA), peptides or polypeptides contained in exosomes, particularly cytoplasmic peptides or proteins, or peptides or polypeptides whose coding RNA is contained in exosomes, are considered to be particularly useful for immunotherapy. Exosomes thus serve as transport vehicles for peptides, proteins, or RNA to antigen-presenting cells, protecting the peptides, proteins, or RNA from degradation by proteases and ribonucleases. Alternatively, peptides and proteins may be taken up by antigen-presenting cells as complexes with other molecules, such as antibodies, via receptor-dependent mechanisms. For example, Tureci et al. (Clin. Cancer Res. 22(8), 1885-1896 (2016)) relate to personalized anti-cancer vaccines utilizing mutation-based neoepitopes. Therefore, in accordance with this understanding, it is a preferred embodiment to particularly select neoantigens previously known in the art as the immune-inducing moiety of the present immunoinducing compound of the present invention.

[0069] In a further preferred embodiment, the TAA is an antigen associated with the cancerous disease in question.

[0070] Examples of potentially useful tumor antigens include cell surface proteins of the claudin family such as p53, ART-4, BAGE, β-catenin / m, Bcr-abL CAMEL, CAP-1, CASP-8, CDC27 / m, CDK4 / m, CEA, CA19-9, CA125, hCG, AFP, claudin-6, claudin-18.2, and claudin-12, c-MYC, CT, Cyp-B, DAM, ELF2M, ETV6-AML1, G250, GAGE, GnT-V, Gap100, HAGE, HER-2 / neu, HPV-E7, HPV-E6, HAST-2, hTERT (or hTRT), LAGE, LDLR / FUT, MAGE-A, preferably MAGE-A1, MAGE-A2, MAGE-A3, MAGE-A4, MAGE-A5, MAGE-A6, MA These are GE-A7, MAGE-A8, MAGE-A9, MAGE-A10, MAGE-A11 or MAGE-A12, MAGE-B, MAGE-C, MART-1 / Melan-A, MC1R, Myosin / m, MUC1, MUM-1, -2, -3, NA88-A, NF1, NY-ESO-1, NY-BR-1, p190 minor BCR-abL, Pm1 / RARa, PRAME, Proteinase 3, PSA, PSM, RAGE, RU1 or RU2, SAGE, SART-1 or SART-3, SCGB3A2, SCP1, SCP2, SCP3, SSX, Survivin, TEL / AML1, TPI / m, TRP-1, TRP-2, TRP-2 / INT2, TPTE, and WT. Particularly preferred tumor antigens include claudin-18.2 (CLDN18.2) and claudin-6 (CLDN6).

[0071] In a further preferred embodiment, the TAA is an antigen associated with a cancer disease different from the cancer disease of interest. The ability to select the immune-inducing moiety according to the desired effect is a particular advantage of the present invention. Therefore, an antigen associated with a cancer disease different from the cancer disease of interest may also be used as the immune-inducing moiety. This could be an antigen known to induce an effective immune response against its respective target cells.

[0072] In a further preferred embodiment, the antigen is a tumor germline antigen, and more preferably, a tumor germline antigen specific to the cancerous disease in question.

[0073] As used herein, the term “tumor germline antigen” preferably refers to proteins or other molecules found on the surface of germline cells, i.e., cells derived from germ cells in the body. These antigens are often recognized by the immune system as non-self or abnormal and can trigger an immune response that helps fight cancer cells. Tumor germline antigens can also be used as targets in cancer immunotherapy, in which the immune system is activated to specifically attack and destroy cancer cells that express these antigens. Thus, tumor germline antigens also constitute a useful immune inducer according to the present invention.

[0074] In a further preferred embodiment, the tumor germline antigen is specific to a second cancerous disease distinct from the cancerous disease of interest.

[0075] With regard to "tumor germline antigens," the inventors hope that it will be understood that the underlying concept of the present invention should not be constrained in the selection of the immune-inducing portion for a specific cancer disease to be treated. Therefore, it is advantageous to be able to select antigens independent of and different from the cancer disease to be treated, particularly tumor germline antigens. Such tumor germline antigens may preferably include angiogenesis markers.

[0076] In a further preferred embodiment, the immune-inducing portion includes an antigen, which is a vaccine antigen and / or a vaccine-associated antigen.

[0077] As used herein, the term “vaccine antigen” preferably refers to a substance used in a vaccine to stimulate an immune response. Also known as an immunogen, this substance is typically a fragment of a pathogenic organism, such as a virus or bacteria, or a protein derived from a pathogen. Vaccine antigens are designed to mimic a pathogen in some way, and when introduced into the body, they trigger the immune system to recognize the antigen and produce antibodies and immune cells that respond to it. Over time, the immune system “remembers” the antigen, enabling a rapid response in the future if the pathogen is encountered again, thereby preventing or mitigating the severity of the disease. This pre-existing memory response is advantageously utilized in the present invention by using a known vaccine antigen as the immune inducer portion, which is also safe and has been particularly well understood, tested, and formally approved for inducing an effective immune response to the vaccine antigen. However, equally important, the use of a vaccine antigen in which the subject has been previously vaccinated increases the effectiveness of the immune response induced by the application of the immune inducer compound (IAC) of the present invention to the subject.

[0078] In a further preferred embodiment, the immune-inducing portion includes an antigen, and the antigen is an exogenous antigen.

[0079] As used herein, the term “exogenous antigen” preferably refers to a substance that does not naturally exist in a host organism and can stimulate an immune response. Exogenous antigens are often used as vaccine antigens because they are less likely to cause harm or adverse reactions and can trigger a stronger and more specific immune response compared to the host’s own antigens. Exogenous antigens can include fragments of pathogens such as viruses or bacteria, and proteins derived from these pathogens. The use of exogenous antigens enables the development of vaccines that can protect against a wide range of diseases, including infectious diseases and cancer. Therefore, the use of exogenous antigens as immune inducers can be particularly advantageous because they are recognized by the immune system as distinct from the host’s own antigens, and as a result can trigger a stronger and more specific immune response, especially if they have been previously used for vaccination. This can lead to the production of more effective antibodies and immune cells that can better protect against the pathogen in question. Furthermore, the use of exogenous antigens reduces the risk of inducing an autoimmune response in which the immune system mistakenly attacks the host’s own tissues. This is because exogenous antigens are less likely to cross-react with the host’s own antigens. Furthermore, exogenous antigens can be used to protect against a wide range of pathogens, including viruses, bacteria, parasites, and cancer cells. This versatility makes it possible to develop vaccines to protect against various diseases. If the subject treated with the IAC of the present invention has previously been vaccinated with such an exogenous antigen, the effectiveness of treatment with each IAC containing the exogenous antigen as the immune-inducing portion can be significantly enhanced. Also, since exogenous antigens do not exist naturally in the body, they are less likely to cause harm or adverse reactions. For this reason, exogenous antigens are a safer option for use compared to using actual pathogens or living attenuated forms of pathogens.

[0080] In a further preferred embodiment, the immune-inducing portion includes an antigen, and the antigen is an immunodominant antigen.

[0081] As used herein, the term “immunodominant antigen” preferably refers to a substance that, when used as a vaccine antigen, induces a potent and dominant immune response. An immunodominant antigen is typically a highly immunogenic protein or other molecule, meaning it can trigger a robust immune response. The term “immunodominant” refers to the fact that this antigen induces a significantly stronger response than the response induced by other antigens in the vaccine or in the pathogen itself. Since immunodominant antigens can better stimulate the immune system to produce antibodies and immune cells that can protect against pathogens, the use of immunodominant antigens in vaccines can lead to the development of more effective vaccines. Therefore, it is immediately apparent that the use of an immunodominant antigen as an immune inducer can similarly induce a strong immune response effective against target cells and / or the target cell environment, mediated by the IAC of the present invention, which contains an immunodominant antigen as an immune inducer.

[0082] In a further preferred embodiment, the immune inducer portion comprises an antigen selected from viral antigens and / or bacterial antigens. Viral and bacterial antigens are substances typically derived from viruses and bacteria, respectively, that can stimulate an immune response. These antigens are used in vaccines to trigger the immune system to produce antibodies and immune cells that can recognize and respond to pathogens. Antigens may be fragments of the pathogen itself or proteins derived from the pathogen. The use of viral and bacterial antigens in vaccines can help protect against a wide range of infectious diseases, including viral infections such as influenza and bacterial infections such as streptococcal pneumonia. By triggering an immune response, these vaccines can help prevent or reduce the severity of the disease and, in some cases, even provide long-term protection against reinfection by the pathogen. Therefore, the use of viral and bacterial antigens as the immune inducer portion of the IAC according to the present invention may be particularly advantageous in using viral and bacterial antigens that are known to induce a strong protective immune response in a subject.

[0083] In a further preferred embodiment, the immune-inducing portion comprises an antigen, and the antigen comprises a B or T cell epitope.

[0084] As used herein, the term “epitope” preferably refers to an antigenic determinant in a molecule such as an antigen, i.e., a portion or fragment of a molecule that is recognized by the immune system, for example, by T cells, when presented in the context of an MHC molecule. Epitopes of proteins such as tumor antigens preferably consist of continuous or discontinuous portions of the protein, preferably 5 to 100 amino acid lengths, preferably 5 to 50 amino acid lengths, more preferably 8 to 30 amino acid lengths, and most preferably 10 to 25 amino acid lengths. For example, an epitope may preferably be 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 amino acid lengths. In the context of the present invention, it is particularly preferable that the epitope is a T cell epitope. According to the present invention, an epitope can bind to MHC molecules, such as MHC molecules on the surface of a cell, and is therefore a "MHC-binding peptide."

[0085] According to the present invention, an epitope can bind to MHC molecules, such as MHC molecules on the surface of a cell, and is therefore a "MHC-binding peptide."

[0086] In a further preferred embodiment, the immune-inducing moiety can induce an antibody and / or TH1 and / or TH2 immune response.

[0087] In a further preferred embodiment, the immune-inducing portion comprises an antigen, which is a viral antigen of a vaccine having a vaccination rate of at least 50%, preferably at least 75%, more preferably at least 90%, and even more preferably at least 95%.

[0088] Those skilled in the art can immediately recognize that vaccination rates for viral antigen-based vaccines in human populations can vary depending on the specific vaccine and population. Generally, vaccination rates for common childhood vaccines in Western countries are very high, often exceeding 90%. For example, according to the World Health Organization (WHO), the vaccination rate for measles, mumps, and rubella (MMR) in the United States is approximately 92%. However, in some Western countries, vaccination rates for certain vaccines, such as the human papillomavirus (HPV) vaccine, which has a vaccination rate of approximately 60-70%, may be lower. Factors that may influence vaccination rates include vaccine availability, vaccination hesitation, and socioeconomic circumstances. Therefore, using an IAC containing an immune-inducing portion that includes the viral antigen of a vaccine with a relatively high vaccination rate will increase the probability of inducing an immune response that has been previously stimulated by that vaccine.

[0089] In a further preferred embodiment, the immune-inducing portion comprises an antigen, which is a viral antigen of a vaccine having an immunization rate of at least 50%, preferably at least 75%, more preferably at least 90%, and even more preferably at least 95%.

[0090] As used herein, the term “immunization rate” is preferably understood as a measure of the proportion of a population that has been vaccinated against a particular disease or has been infected with a particular disease, and therefore has a memory response, particularly a B-cell response, to said vaccination / infection. Immunization rates are usually expressed as a percentage and reflect the number of individuals in a population that have received all therapeutic units of a vaccine and / or have experienced infection with the respective pathogen. Immunization rates are an important evaluation criterion in the field of public health because they provide an indicator of the level of protection provided to a population against a particular disease. High immunization rates are often associated with lower incidences of disease and can help prevent epidemics and reduce the spread of infectious diseases. To achieve and maintain high immunization rates, it is important to have a strong public health infrastructure and an effective vaccine delivery system, as well as broad public awareness and acceptance of the benefits of vaccination. If such antigens of a vaccine having a relatively high immunization rate are selected as the immune-inducing portion of the IAC of the present invention, this favorably increases the probability of inducing a strong immune response against target cells and / or the target cell environment in the subject, and thus favorably increases the effectiveness of treating the cancerous disease of the subject with the IAC.

[0091] In a further preferred embodiment, the immune-inducing portion comprises an antigen selected from or derived from measles, mumps, rubella (MMR) vaccine, diphtheria, tetanus, polio (DTP) vaccine, Haemophilus influenzae type b (Hib) vaccine, pneumococcal vaccine, hepatitis B vaccine, or SARS-CoV-19 vaccine, or a portion thereof.

[0092] In most countries, especially developed countries, there are several mandatory vaccinations aimed at protecting populations, and sometimes children in particular, from certain diseases. These vaccinations are either part of a recommended vaccination schedule or required by law.

[0093] Mandatory childhood vaccinations in Germany include, for example, measles, mumps, rubella (MMR), diphtheria, tetanus, polio (DTP), Haemophilus influenzae type b (Hib), pneumococcal, or hepatitis B. The measles, mumps, and rubella (MMR) vaccine protects against three diseases and is usually administered in two doses at ages 11-14 months and 15-23 months. The MMR vaccine contains live, attenuated strains of the viruses that cause measles, mumps, and rubella. These viruses are used as antigens in the vaccine. The diphtheria, tetanus, and polio (DTP) vaccine protects against diphtheria, tetanus, and polio and is usually administered in four doses at ages 2, 3, 4, and 11-14 months. DTP vaccines contain inactivated (dead) forms of the toxins that cause diphtheria and tetanus, and small amounts of inactivated poliovirus. These toxins and viruses are used as antigens in the vaccine. Haemophilus influenzae type b (Hib) vaccination protects against bacterial infections that can cause serious complications such as meningitis or sepsis. Hib vaccines are usually given in three doses at ages 2, 4, and 12 months. Hib vaccines contain a protein called polyribosylribitol phosphate (PRP) extracted from the outer surface of Haemophilus influenzae type b bacteria, and this protein is used as an antigen in the vaccine. Pneumococcal vaccination protects against bacterial infections that can cause serious complications such as pneumonia or meningitis. Pneumococcal vaccines are usually given in three doses at ages 2, 4, and 12 months. Pneumococcal vaccines contain proteins called polysaccharides, which are extracted from the surface of certain types of Streptococcus pneumoniae bacteria, and these polysaccharides are used as antigens in the vaccine. Hepatitis B vaccination protects against viral infections that can cause liver disease. Hepatitis B vaccine is usually given in three doses at ages 2, 4, and 12 months.The hepatitis B vaccine contains a protein called hepatitis B surface antigen (HBsAg), which is extracted from the surface of the hepatitis B virus, and this protein is used as an antigen in the vaccine.

[0094] However, there are some vaccinations that are not mandatory but are recommended, such as influenza, meningococcal, and tick-borne encephalitis vaccinations. It is always important to discuss vaccination with your doctor or pediatrician and to be familiar with the recommendations and regulations regarding vaccination in Germany.

[0095] Recently, although not considered mandatory in many countries, SARS-CoV-19 vaccination programs have also achieved high immunization rates for SARS-CoV-19 in the population.

[0096] Therefore, the use of antigens selected from or derived from measles, mumps, rubella (MMR) vaccines, diphtheria, tetanus, polio (DTP) vaccines, Haemophilus influenzae type b (Hib) vaccine, pneumococcal vaccine, hepatitis B vaccine, and SARS-CoV-19 vaccine as the immune-inducing portion of the IAC according to the present invention may be particularly advantageous because the probability that the subject to be treated has already experienced a primary immune response to the said vaccine antigens is considerably increased. Thus, the possibility of a stronger immune response mediated by the application of the IAC according to the present invention, and therefore, almost certainly, a more effective treatment of the cancerous disease of the subject, is increased.

[0097] In a further preferred embodiment, the immune-inducing portion comprises an antigen selected from the group including natural substances, antibodies, affilins, peptides, proteins, carbohydrates, lipids, nucleic acids, synthetic compounds or toxoids, or combinations thereof.

[0098] Peptides and proteins are common antigens because they are major components of many pathogens, such as viruses and bacteria, and are recognized as non-self by the immune system. Carbohydrates, lipids, and nucleic acids are also used as antigens because they are not present in the host and can provide a distinctive molecular signature that can be recognized as non-self by the immune system. Synthetic compounds and toxoids are also used as antigens because they can mimic the structure of pathogen-associated antigens and induce similar immune responses. Thus, some viruses, such as the influenza virus, are thought to have carbohydrate antigens on their surface. Furthermore, some bacteria, such as Mycobacterium tuberculosis, have lipid antigens that are recognized by the immune system. Some viruses, such as human immunodeficiency virus (HIV), have nucleic acid antigens that are recognized by the immune system. Some vaccines, such as toxoid vaccines, use synthetic compounds as antigens. Some toxoid vaccines, such as those for diphtheria and tetanus, use inactivated toxins produced by bacteria as antigens. It is also worth noting that vaccines can use combinations of different types of antigens to produce a more robust immune response. Therefore, in one embodiment of the present invention, it is considered that a combination of at least two different antigens is included in the immune-inducing portion of the IAC according to the present invention. Thus, these types of antigens, as described above, included in the immune-inducing portion of the IAC of the present invention as a single antigen or multiple antigens, may be particularly advantageous in effectively inducing an immune response due to their ability to trigger the immune system and produce an immune response without requiring nucleic acid translation.

[0099] In a further preferred embodiment, the immune-inducing moiety (3) comprises an antigen, which is a synthetic compound selected from the group comprising small molecules.

[0100] In a further preferred embodiment, the immune-inducing portion (3) comprises an antigen, the antigen being a natural substance, in particular a plant-derived antigen such as cannabidiol (CBD), or any other natural substance that can stimulate an immune response in a subject.

[0101] As used herein, the term “natural substance” preferably refers to a naturally occurring, preferably as-is, chemical compound or material. In other words, natural substances are preferably not significantly altered or synthesized by human processes. These natural substances are typically derived from plants, animals, minerals, or microorganisms. Natural substances can encompass a wide range of compounds, including minerals, vitamins, proteins, carbohydrates, lipids, and various organic molecules.

[0102] In a further preferred embodiment, the immune-inducing moiety comprises a nucleic acid selected from the group including DNA, RNA, mRNA, rRNA, tRNA, miRNA, siRNA, snRNA, piRNA, and lncRNA. The use of nucleic acids, particularly DNA and RNA, particularly mRNA, as the immune-inducing moiety is especially advantageous because nucleic acids can encode genetic information and are readily expressed within cells, making them excellent carriers for antigens. This makes nucleic acids ideal for use in vaccines, as they can be used to deliver antigen information to cells and trigger an immune response. One approach to using nucleic acids as carriers for antigens is through the use of DNA vaccines. In this approach, a plasmid encoding the antigen is delivered into cells where the antigen is expressed and processed to present the antigen to the immune system. An immune response to the antigen is then triggered, leading to the development of immunological memory that will enable the recognition and response to pathogens in the future. DNA vaccines have several advantages, including ease of manufacture, stability, and the ability to be delivered using various administration routes, such as intramuscular injection or intradermal administration. The use of each DNA molecule in a DNA vaccine is therefore particularly advantageous when that DNA molecule is used as an immune-inducing portion of the IAC of the present invention or as part of an immune-inducing portion of the IAC of the present invention. Thereafter, those skilled in the art will immediately understand that nucleic acid molecules can directly induce immune cells or require internal translocation and translation, accompanied by the subsequent presentation of the resulting peptides or proteins on the cell surface of cells, particularly APCs. Thereafter, the underlying idea of ​​the inventors is that in tumor cells or tumor cell environments, particularly when irradiated with radionuclides, which may even be contained in the IAC of the present invention in some embodiments, the destroyed cells and their respective fragments are taken up by immune cells such as macrophages and subsequently presented to further immune cells. By the same mechanism, the nucleic acid of the immune-inducing portion is taken up, ultimately translated, and presented either by itself or in the form of a translation product or part thereof, such as a peptide.

[0103] Therefore, the use of RNA vaccines that utilize messenger RNA (mRNA) as a carrier for antigen information is also conceivable in the context of this invention. mRNA is taken up by cells, where it is translated into proteins, and then the antigen is presented to the immune system. RNA vaccines have several advantages over conventional protein-based vaccines, including the ability to rapidly produce large quantities of vaccine, to respond quickly to new threats, and to design vaccines against rapidly evolving pathogens such as the SARS-CoV-2 virus that causes COVID-19. In summary, nucleic acids are excellent carriers for antigens because they can deliver antigen information to cells, triggering antigen expression and an immune response. For this reason, nucleic acids represent a promising approach to developing new vaccines for a wide range of infectious diseases and to improving existing vaccines.

[0104] The approach of using mRNA molecules as antigens may, in particular, derive from International Publication No. 2021 / 213924, which is considered to be incorporated herein by reference. At the very least, those skilled in the art will have sufficient knowledge to incorporate the findings and embodiments of International Publication No. 2021 / 213924 when drafting the IAC according to the present invention.

[0105] In preferred embodiments of the IAC of the present invention, the immune-inducing moiety comprises an mRNA molecule encoding an amino acid sequence containing the SARS-CoV-2 S protein and / or an immunogenic variant thereof, or an immunogenic fragment of the SARS-CoV-2 S protein or an immunogenic variant thereof.

[0106] The antigens in COVID-19 vaccines used during the pandemic years of 2020–2023 are small pieces of genetic material called messenger RNA (mRNA) that contain instructions for producing the spike protein of the SARS-CoV-2 virus, as described above. The mRNA is packed into lipid nanoparticles that protect the mRNA and help deliver it to cells. Once inside the cell, the mRNA is used as a template for making the spike protein, which then protrudes from the cell surface.

[0107] In a preferred embodiment of the IAC of the present invention, the immune-inducing portion comprises nucleic acid molecules packed within lipid nanoparticles.

[0108] In a further preferred embodiment, the immune-inducing moiety comprises an immunogenic fragment of the SARS-CoV-2 S protein, and includes the S1 subunit of the SARS-CoV-2 S protein, or the receptor-binding domain (RBD) of the SARS-CoV-2 S protein, preferably the S1 subunit of the SARS-CoV-2 S protein. The receptor-binding domain (RBD) of the SARS-CoV-2 S protein preferably has SEQ ID NO: 8.

[0109] The receptor-binding domain (RBD) in the SARS-CoV-2 S protein was specifically characterized by Wanbo Tai et al. (Tai, W., He, L., Zhang, X. et al. Characterization of the receptor-binding domain (RBD) of 2019 novel coronavirus: implication for development of RBD protein as a viral attachment inhibitor and vaccine. Cell Mol Immunol 17, 613-620 (2020). https: / / doi.org / 10.1038 / s41423-020-0400-4).

[0110] In particular, the receptor-binding domain (RBD) of the S1 subunit of the SARS-CoV-2 S protein corresponds to a critical component in the interaction between the virus and host cells; therefore, using the receptor-binding domain (RBD) of the S1 subunit of the SARS-CoV-2 S protein may be advantageous in the context of the present invention. Accordingly, using the RBD as part of or as an immune-inducing portion in the IAC according to the present invention may have certain advantages. In particular, using the RBD may potentially enhance the effectiveness of the IAC according to the present invention by utilizing an immune system effectively trained to recognize and respond to a critical functional portion of the virus, thereby leveraging the immune response to SARS-CoV-2 in the target, preferably an existing immune response.

[0111] In a further preferred embodiment, the amino acid sequence comprising the SARS-CoV-2 S protein, its immunogenic mutant, or an immunogenic fragment of the SARS-CoV-2 S protein or its immunogenic mutant is encoded by a coding sequence that is codon-optimized and / or has an increased G / C content compared to the wild-type coding sequence, wherein the codon optimization and / or increased G / C content preferably does not alter the sequence of the encoded amino acid sequence.

[0112] Codon optimization and increased G / C content in the coding sequence are particularly advantageous because they enhance expression efficiency without altering the amino acid sequence, ensuring optimal protein production. As a result, this modification contributes to the development of improved vaccines or treatments by maximizing the expression and immunogenicity potential of the SARS-CoV-2 S protein or its variants.

[0113] In a particularly preferred embodiment of the IAC of the present invention, the immune inducer portion is (i) Preferably, a nucleic acid molecule, particularly an RNA molecule, encoding the SARS-CoV-2 S protein, an immunogenic variant thereof, or an immunogenic fragment of the SARS-CoV-2 S protein or an immunogenic variant thereof, comprising the nucleotide sequence of nucleotides 979-1584 of SEQ ID NO: 1, 2, or 3, a nucleotide sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the nucleotide sequence of nucleotides 979-1584 of SEQ ID NO: 1, 2, or 3, or a fragment of a nucleotide sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the nucleotide sequence of nucleotides 979-1584 of SEQ ID NO: 1, 2, or 3; and / or (ii) Preferably, an immunogenic fragment of the SARS-CoV-2 S protein, an immunogenic variant thereof, or an immunogenic fragment of the SARS-CoV-2 S protein or an immunogenic variant thereof, comprising the amino acid sequence of amino acids 327-528 of SEQ ID NO: 4, an amino acid sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the amino acid sequence of amino acids 327-528 of SEQ ID NO: 4, or an immunogenic fragment of the amino acid sequence of amino acids 327-528 of SEQ ID NO: 4, having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the amino acid sequence of amino acids 327-528 of SEQ ID NO: 4; and / or (iii) Preferably, an immunogenic fragment of the SARS-CoV-2 S protein RBD, an immunogenic variant thereof, or an immunogenic fragment of the SARS-CoV-2 S protein RBD or an immunogenic variant thereof, comprising the amino acid sequence of SEQ ID NO: 8, an amino acid sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the amino acid sequence of SEQ ID NO: 8, or an immunogenic fragment of an amino acid sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the amino acid sequence of SEQ ID NO: 8 Includes.

[0114] In a particularly preferred embodiment of the IAC of the present invention, the immune inducer portion is (i) A nucleic acid molecule, particularly an RNA molecule, encoding the SARS-CoV-2 S protein, its immunogenic variant, or an immunogenic fragment of the SARS-CoV-2 S protein or its immunogenic variant, comprising the nucleotide sequence 49-2055 of SEQ ID NO: 1, 2, or 3, a nucleotide sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the nucleotide sequence 49-2055 of SEQ ID NO: 1, 2, or 3, or a fragment of a nucleotide sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the nucleotide sequence 49-2055 of SEQ ID NO: 1, 2, or 3; and / or (ii) Preferably, an immunogenic fragment of the SARS-CoV-2 S protein, an immunogenic variant thereof, or an immunogenic fragment of the SARS-CoV-2 S protein or an immunogenic variant thereof, comprising the amino acid sequence of amino acids 17-685 of SEQ ID NO: 4, an amino acid sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the amino acid sequence of amino acids 17-685 of SEQ ID NO: 4, or an immunogenic fragment of the amino acid sequence of amino acids 17-685 of SEQ ID NO: 4, having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the amino acid sequence of amino acids 17-685 of SEQ ID NO: 4; and / or (iii) Preferably, an immunogenic fragment of the SARS-CoV-2 S protein RBD, an immunogenic variant thereof, or an immunogenic fragment of the SARS-CoV-2 S protein RBD or an immunogenic variant thereof, comprising the amino acid sequence of SEQ ID NO: 8, an amino acid sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the amino acid sequence of SEQ ID NO: 8, or an immunogenic fragment of an amino acid sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the amino acid sequence of SEQ ID NO: 8 Includes.

[0115] In a further preferred embodiment, (i) a nucleic acid molecule, particularly an RNA molecule, encoding the SARS-CoV-2 S protein, its immunogenic variant, or an immunogenic fragment of the SARS-CoV-2 S protein or its immunogenic variant, is a nucleotide sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the nucleotide sequence of nucleotides 49-3819 of SEQ ID NO: 1, 2, or 3, or at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the nucleotide sequence of nucleotides 49-3819 of SEQ ID NO: 1, 2, or 3 or at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the nucleotide sequence of nucleotides 49-3819 of SEQ ID NO: 1, 2, or 3 (ii) preferably an immunogenic fragment of the SARS-CoV-2 S protein, an immunogenic variant thereof, or an immunogenic fragment of the SARS-CoV-2 S protein or an immunogenic variant thereof, comprising: an amino acid sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the amino acid sequence of amino acids 17-1273 of SEQ ID NO: 4 or 5; or an immunogenic fragment of the amino acid sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the amino acid sequence of amino acids 17-1273 of SEQ ID NO: 4 or 5;And / or preferably, an immunogenic fragment of the SARS-CoV-2 S protein, an immunogenic variant thereof, or an immunogenic fragment of the SARS-CoV-2 S protein, comprising the amino acid sequence of SEQ ID NO: 8, an amino acid sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the amino acid sequence of SEQ ID NO: 8, or an immunogenic fragment of an amino acid sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the amino acid sequence of SEQ ID NO: 8.

[0116] In a further preferred embodiment, the amino acid sequence comprising the SARS-CoV-2 S protein, its immunogenic mutant, or an immunogenic fragment of the SARS-CoV-2 S protein or its immunogenic mutant comprises a secretory signaling peptide.

[0117] The inclusion of secreted signaling peptides is particularly advantageous because it ensures efficient extracellular exposure, promotes enhanced antigen presentation to immune cells, and potentially improves the effectiveness of SARS-CoV-2 vaccine or immunotherapy strategies.

[0118] In a further preferred embodiment, the secretory signaling peptide is preferably fused at the N-terminus to the SARS-CoV-2 S protein, its immunogenic mutant, or an immunogenic fragment of the SARS-CoV-2 S protein or its immunogenic mutant.

[0119] In a further preferred embodiment, (i) the RNA encoding the secretion signal peptide is a nucleotide sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the nucleotide sequence of nucleotides 1-48 of SEQ ID NO: 1, 2, or 3, or at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, or at least 85. (ii) the secretory signal peptide comprises an amino acid sequence of amino acids 1-16 of SEQ ID NO: 4, an amino acid sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity to the amino acid sequence of amino acids 1-16 of SEQ ID NO: 4, or a functional fragment of an amino acid sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity to the amino acid sequence of amino acids 1-16 of SEQ ID NO: 4.

[0120] In a further preferred embodiment, (i) The RNA encoding the SARS-CoV-2 S protein, its immunogenic variant, or an immunogenic fragment of the SARS-CoV-2 S protein or its immunogenic variant, comprises the nucleotide sequence of SEQ ID NO: 6, a nucleotide sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the nucleotide sequence of SEQ ID NO: 6, or a fragment of a nucleotide sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the nucleotide sequence of SEQ ID NO: 6; and / or (ii) the SARS-CoV-2 S protein, its immunogenic variant, or SARS-CoV-2 The immunogenic fragments of the S protein or its immunogenic variant include the amino acid sequence of SEQ ID NO: 7, an amino acid sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the amino acid sequence of SEQ ID NO: 7, or an immunogenic fragment of an amino acid sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the amino acid sequence of SEQ ID NO: 7.

[0121] In a further preferred embodiment, the immune-inducing moiety comprises a peptide having a length of at least 8 amino acids, preferably at least 10 amino acids, and more preferably at least 15 amino acids.

[0122] According to the present invention, the term "peptide" preferably refers to a short chain of amino acids linked by peptide bonds. Peptides are building blocks of proteins and are used in a variety of biological processes, including signal transduction, regulatory, and structural roles. In relation to vaccines, peptides can be used as antigens, which are substances that trigger an immune response in the body. Peptides can be designed to mimic specific regions of pathogen proteins, providing a target for the immune system to respond to. By inducing an immune response to peptide antigens, it is possible to develop immunity to pathogens and protect against infection. The definition of a peptide in a patent application may also include details about the structure and properties of the peptide, such as the number of amino acids, a specific amino acid sequence, and methods of synthesis and production. In the context of the present invention, the term "peptide" also refers to a substance containing two or more, preferably three or more, preferably four or more, preferably six or more, preferably eight or more, preferably ten or more, preferably thirteen or more, preferably sixteen or more, preferably twenty-one or more, and most preferably eight, ten, twenty-five, or fifty, and especially 100 amino acids, which are covalently linked by peptide bonds. The terms "polypeptide" or "protein" refer to a larger peptide, preferably a peptide having more than 100 amino acid residues, but generally the terms "peptide," "polypeptide," and "protein" are synonymous and are used interchangeably herein.

[0123] In a further preferred embodiment, the immune-inducing moiety comprises a peptide having a maximum length of 200 amino acids, preferably 150 amino acids, more preferably 100 amino acids, even more preferably 65 amino acids, even more preferably 60 amino acids, and even more preferably 25 amino acids.

[0124] Those skilled in the art will recognize that, particularly in vaccination, the length of peptides used to induce an immune response can vary depending on the specific vaccine and target antigen. Peptide vaccines typically use short synthetic peptides, usually 15–30 amino acids long, that mimic small portions of the antigen. This is because these short peptides are easier to synthesize in the laboratory and are more likely to be recognized by the immune system. However, some vaccines use longer peptides, up to 60 amino acids long, to mimic more complex regions of the antigen. Furthermore, some other vaccines use full-length proteins as antigens.

[0125] The terms “part” and “fragment” or “part thereof” are used interchangeably herein and refer to a contiguous element. For example, a part of a structure such as an amino acid sequence or a protein refers to a contiguous element of the said structure. A part, portion, or fragment of a structure preferably includes one or more functional properties of the said structure. For example, a part, portion, or fragment of an epitope, peptide, or protein preferably is immunologically equivalent to the epitope, peptide, or protein from which it is derived. In the context of the present invention, "part" of a structure such as an amino acid sequence preferably includes at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 92%, at least 94%, at least 96%, at least 98%, and at least 99% of the whole structure or amino acid sequence, and preferably consists of at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 92%, at least 94%, at least 96%, at least 98%, and at least 99% of the whole structure or amino acid sequence.

[0126] In a further preferred embodiment, the immune-inducing portion includes an antigen that is a subunit, a recombinant or conjugated vaccine or part thereof, or an antigen derived from a subunit, a recombinant or conjugated vaccine or part thereof.

[0127] In the case of subunit, recombinant, or conjugate vaccines, the antigen is a specific part or component of the virus or bacterium against which the vaccine is designed to provide protection. For example, in the case of Haemophilus influenzae type b (Hib) vaccine, the antigen is the bacterial polysaccharide capsule. This capsule is a surface component of the bacterium recognized by the immune system and is therefore an effective antigen for use in the vaccine. In the case of human papillomavirus (HPV) vaccine, the antigen is a specific protein called L1, which is a structural protein of the virus. In subunit vaccines, the antigen is a purified protein or a specific part of the pathogen.

[0128] In recombinant vaccines, the antigen is a genetically modified version of a pathogen, usually a specific protein derived from the pathogen. In conjugate vaccines, the antigen is a combination of a pathogen protein and a protein from another organism, which helps to enhance the immune response.

[0129] These subunits, recombinants, and conjugate vaccines are designed to induce an immune response against specific parts of a pathogen and are beneficial for protection against certain infections. Therefore, immune-inducing moieties containing subunits, recombinants, or conjugate vaccines or antigens that are parts thereof, or antigens derived from subunits, recombinants, or conjugate vaccines or parts thereof, may be particularly advantageous in inducing a strong immune response against target cells and in the treatment of targets with IACs of the present invention.

[0130] In a further preferred embodiment, the immune-inducing moiety comprises an antigen that is a toxoid or a part thereof, or an antigen derived from a toxoid or a part thereof, or the immune-inducing moiety comprises an antigen derived from a toxoid and expressed by target cells. In the case of toxoid vaccines, the antigen is a modified or inactivated form of a toxin (toxic substance) produced by a particular bacterium. Toxoids are used to create vaccines against diseases caused by toxin-producing bacteria, such as tetanus and diphtheria. For example, in a tetanus vaccine, the antigen is an inactivated form of tetanus toxin. The inactivation process prevents the toxin from causing disease, but it can still stimulate the immune system to produce an immune response. In the case of a diphtheria vaccine, the antigen is an inactivated form of diphtheria toxin. By exposing the immune system to these inactivated forms of toxins, the immune system can learn to recognize and respond to these toxins if a person is exposed to them in the future. These toxoid vaccines are highly effective in providing long-term immunity against these diseases. Therefore, an immune-inducing moiety containing a toxoid or an antigen that is a part thereof, or an antigen derived from a toxoid or a part thereof, may be particularly advantageous in inducing a strong immune response against target cells and in the treatment of a target by the IAC of the present invention.

[0131] In a further preferred embodiment, the immune-inducing moiety includes a protein derived from the SARS-CoV-19 virus, particularly the SARS-CoV-19 spike protein, or an antigen which is a part thereof.

[0132] The immune system recognizes the spike protein as non-self and initiates an immune response against it. This immune response can then protect the person from becoming infected with the virus if they are exposed to it in the future.

[0133] Currently, there are several different COVID-19 vaccines authorized for emergency use by regulatory bodies worldwide, including Pfizer-BioNTech, Moderna, AstraZeneca, and Johnson & Johnson. These vaccines use different technologies such as mRNA, vector systems, and inactivated viruses, but all of them target the SARS-CoV-2 virus spike protein, which is the antigen used to stimulate the immune response. Of course, as considered herein, the antigen contained in the immune-inducing portion of the IAC of the present invention may be a peptide of the SARS-CoV-19 spike protein, the whole protein, or a part thereof.

[0134] In further, particularly preferred embodiments, the immune-inducing moiety comprises an antigen, which is an RBD protein derived from the SARS-CoV-19 virus. The receptor-binding domain (RBD) of the SARS-CoV-2 S protein preferably has SEQ ID NO: 8.

[0135] In a further preferred embodiment, the immune-inducing portion comprises a nucleic acid selected from RNA, DNA, or a combination thereof. It is known in the art that antigens can be encoded by both DNA and RNA, which are two major types of nucleic acids. Such antigens may be particularly advantageous if they are included in the immune-inducing portion of the IAC of the present invention. DNA-based antigens are typically used in DNA vaccines. In these vaccines, a small piece of DNA containing genetic information about a specific antigen is delivered into the body. Once inside the cell, the DNA is used as a template for producing an antigen protein. The immune system then recognizes the antigen protein as non-self and initiates an immune response against it. RNA-based antigens are typically used in RNA vaccines. In these vaccines, a small piece of RNA called messenger RNA (mRNA) containing genetic information about a specific antigen is delivered into the body. Once inside the cell, the mRNA is used as a template for producing an antigen protein. The immune system then recognizes the antigen protein as non-self and initiates an immune response against it. Currently, the most common COVID-19 vaccine is an mRNA vaccine, which, as mentioned above, uses a small fragment of mRNA encoding the spike protein of the SARS-CoV-2 virus, and this is a new technology. Both DNA vaccines and RNA vaccines have the potential to provide immunity against certain infectious diseases, but RNA vaccines have the advantage of being easier to produce and handle than DNA vaccines. In either case, the immune-inducing moiety, which includes a nucleic acid selected from RNA, DNA, or a combination thereof, may be advantageous in inducing an immune response against target cells and / or the target cell environment when using the IAC of the present invention in the treatment of cancerous diseases. In particular, in embodiments in which the IAC further includes a radionuclide, the target cells and / or the target cell environment may be damaged by irradiation, and each immune-inducing moiety may be taken up by immune cells, resulting in the presentation of nucleic acids or translation products or parts thereof that carry the immune cell attractant.Therefore, a further specific advantage of using nucleic acids as or as part of an immune attractant is to prevent or reduce the effect that IACs can be recognized by antibodies present in the blood, which can, to some extent, lead to the elimination of IACs. This effect can be advantageously reduced in that the immune attractant comprises nucleic acids selected from RNA or DNA or a combination thereof, and the nucleic acids present antigens that ultimately attract immune cells to target cells and / or the target cell environment only when taken up by immune cells.

[0136] In a further preferred embodiment, the immune-inducing portion comprises tetanus toxin or a portion thereof. The antigen in the tetanus vaccine is a protein called tetanus toxin. Tetanus toxin is produced by the bacterium Clostridium tetani, which is found in soil, dust, and fertilizer. This toxin can enter the body through a puncture wound or other wound in the skin, travel to the central nervous system, where it blocks the release of neurotransmitters, resulting in muscle rigidity and spasms. Tetanus vaccine is usually given as part of a combination vaccine called tetanus, diphtheria, and pertussis (Tdap) vaccine. The Tdap vaccine contains inactivated (dead) tetanus toxoid, which is a form of tetanus toxin that has been processed to be non-toxic but still capable of stimulating an immune response. This vaccine is administered to help the body develop immunity to tetanus toxin so that, if a person is exposed to toxin-producing bacteria, the immune system can quickly and effectively neutralize the toxin, preventing the development of tetanus. Receiving a tetanus vaccine is important because tetanus is a serious disease that can cause severe physical disability and even death, especially in the elderly. This vaccine is usually administered in a series of doses, with booster doses given periodically to maintain immunity. Therefore, an immune-inducing moiety containing tetanus toxin or a portion thereof may be particularly advantageous in inducing a strong immune response against target cells and in the treatment of targets with the IAC of the present invention.

[0137] In a further preferred embodiment, the immune-inducing moiety comprises the hemagglutinin (HA) protein or a portion thereof. The antigen in the influenza (flu) vaccine is a viral protein called the hemagglutinin (HA) protein. Influenza viruses are constantly changing, and therefore it is important to update the vaccine annually to protect against the latest strains. This vaccine consists of inactivated or killed influenza viruses, or viral subunits containing the HA protein. This vaccine is designed to protect against the most common strains of influenza that are expected to be prevalent during a given influenza season. This vaccine is typically administered in the fall, before the start of the influenza season, and is recommended for almost everyone over 6 months of age, especially infants, pregnant women, people with certain chronic medical conditions, and the elderly, who are at high risk of severe influenza complications.

[0138] Therefore, an immune-inducing moiety containing hemagglutinin (HA) protein or a portion thereof may be particularly advantageous in inducing a strong immune response against target cells and in the treatment of targets with IAC according to the present invention.

[0139] In a further preferred embodiment, the immune-inducing moiety comprises polyribosylribitol phosphate (PRP). Polyribosylribitol phosphate (PRP) is a type of carbohydrate polymer found in the outer membrane of some bacteria, such as Haemophilus influenzae type b (Hib). PRP has been used in several vaccines as an antigen or immune system target to provide protection against bacteria. PRP-based vaccines are widely used and have been shown to be effective and safe. The use of PRP as an antigen in vaccines allows for the generation of immunity without causing disease, providing a method for protection against dangerous bacteria in a safe and controlled manner.

[0140] Therefore, the immune-inducing portion containing polyribosylribitol phosphate (PRP) may be particularly advantageous in inducing a strong immune response against target cells and in the treatment of the target by the IAC of the present invention.

[0141] In a further preferred embodiment, the immune-inducing moiety comprises at least one surface polysaccharide of Streptococcus pneumoniae. Surface polysaccharides of Streptococcus pneumoniae (Pneumococcal) are a type of carbohydrate found on the outer surface of the pneumococcal bacterium. In pneumococcal disease, surface polysaccharides can be used as antigens in vaccines to provide protection against pneumococcal infections. The vaccine works by exposing the immune system to the surface polysaccharide, which is then recognized by the body, generating an immune response. This response results in the production of antibodies that can protect against future infections by pneumococcal bacteria. Pneumococcal polysaccharide vaccines (PPVs) are widely used to protect against pneumococcal infections such as pneumococcal pneumonia, meningitis, and sepsis. PPVs are recommended for individuals at increased risk of pneumococcal infections, including the elderly, people with certain chronic medical conditions, and those with weakened immune systems. PPV is considered effective and safe and has been shown to reduce the incidence of pneumococcal disease. The use of surface polysaccharides as antigens in vaccines allows for the generation of immunity without causing disease, providing a method for protection against pneumococcal infection in a safe and controlled manner. Therefore, an immune-inducing moiety containing at least one surface polysaccharide of Streptococcus pneumoniae may be particularly advantageous in inducing a strong immune response against target cells and in the treatment of targets with IAC of the present invention.

[0142] In a further preferred embodiment, the immune-inducing moiety comprises hepatitis B surface antigen (HBsAg). HBsAg is a protein found on the surface of the hepatitis B virus (HBV). HBsAg is one of the earliest markers of HBV infection, and its presence in the blood indicates that a person is infected with the virus. HBsAg is also used as an antigen in hepatitis B vaccines. This vaccine works by exposing the immune system to HBsAg, which is then recognized by the body, generating an immune response. This response results in the production of antibodies that can protect against future infections by HBV. HBsAg-based vaccines are widely used and have been shown to be very effective in preventing HBV infection. The use of HBsAg as an antigen in vaccines allows for the generation of immunity without causing disease, providing a method for protection from HBV in a safe and controlled manner. HBsAg-based vaccines are recommended for all newborns, children, and adolescents, as well as for certain high-risk groups such as healthcare workers, people with multiple sexual partners, people who inject drugs, and individuals with chronic liver disease. HBsAg-based vaccines have helped reduce the incidence of HBV infection and its associated morbidity and mortality worldwide. Therefore, an immune-inducing moiety containing hepatitis B surface antigen (HBsAg) may be particularly advantageous in inducing a strong immune response against target cells and in the treatment of targets with the IAC of the present invention.

[0143] In a further preferred embodiment, the immunoattractant compound (IAC) comprises more than one, preferably two, immunoattractant moieties.

[0144] This is particularly advantageous because the target immune system can be stimulated in parallel by two immune-inducing moieties. These can be one or more, preferably two, identical immune-inducing moieties, or one or more, preferably two, different immune-inducing moieties. In particular, the use of one or more immune-inducing moieties can increase the effect on the immune system, the likelihood of immune cells binding to their immune cell receptors, and / or the affinity of immune cells to their immune cell receptors. When different immune-inducing moieties are used, different initial antigen-stimulating effects on the target, such as the immune response to a previous vaccination or infection event, can be advantageously utilized, and the likelihood of the target having a corresponding memory response to at least one of the different immune-inducing moieties is advantageously increased.

[0145] In a further preferred embodiment, the target cells are cancer cells, and the disease is a cancerous disease. According to the present invention, “tumor” is used herein to refer to an abnormal mass of tissue which may be benign (non-cancerous) or malignant (cancerous). Benign tumors do not spread to other parts of the body and do not pose a serious threat to health. On the other hand, malignant tumors can invade nearby tissues and spread to other parts of the body via the bloodstream or lymphatic system, which is known as metastasis. On the other hand, cancer is used herein to refer to a group of diseases which are preferably characterized by the uncontrolled proliferation and spread of abnormal cells. Cancer cells divide and proliferate in an uncontrolled manner and can invade and damage nearby tissues and organs. Cancer can also spread to other parts of the body via the bloodstream or lymphatic system.

[0146] The IAC of the present invention is particularly suitable for the treatment of cancerous diseases. Here, the presence and / or overexpression of a target structure indicates the target cancerous disease. Thus, the target cell environment of the target cells and / or target call may or may not be cancerous cells. It is particularly preferable that the target cells are cancerous cells and the disease is a cancerous disease. In such embodiments, the IAC directly targets cancer cells and directly transmits its effective treatment to the root cause of the disease. However, it may be considered as an alternative that non-cancer cells, particularly immune cells, are also targeted, thereby stimulating the immune system and enabling targeting of tumor-specific antigens. In such embodiments, care must be taken to avoid causing undesirable damage to the target body, but the inventors have also considered use in which the immune system is stimulated and "heated up" away from the scene, i.e., not in an immunosuppressive tumor environment. In this way, if TAA is selected as particularly included in the immune-inducing portion, the immune system can be stimulated by the IAC of the present invention to specifically induce an immune response against cancer, without being hindered by TIME.

[0147] In a further preferred embodiment, the disease is a cancer such as carcinoma, sarcoma, leukemia, lymphoma, melanoma, brain tumor, or other disease involving abnormal cell proliferation.

[0148] Carcinoma is the most common type of cancer and arises in the epithelial cells that cover the surface of internal organs and skin. Examples include lung cancer, breast cancer, and colon cancer. Sarcoma is understood as cancer that begins in bone, muscle, tendon, and other connective tissue. Examples include osteosarcoma and synovial sarcoma. Leukemia is cancer that begins in the hematopoietic cells of the bone marrow. Examples include acute lymphoblastic leukemia and chronic myeloid leukemia. Lymphoma is cancer that begins in the lymphatic system, which is a network of tubes and organs that help fight infection. Examples include Hodgkin lymphoma and non-Hodgkin lymphoma. Melanoma is cancer that begins in melanin-producing cells, which are cells that produce the pigment that gives color to the skin. Brain cancer is cancer that begins in the brain and can be benign or malignant. Among malignant neoplasms, adenocarcinoma begins in glandular (secretory) cells. Examples include lung adenocarcinoma, prostate adenocarcinoma, and colon adenocarcinoma. Squamous cell carcinoma is a type of carcinoma that originates in squamous epithelial cells, which are the thin, flat cells that cover the surface of internal organs and skin. Examples include squamous cell carcinoma of the lung, squamous cell carcinoma of the head and neck, and squamous cell carcinoma of the cervix. Transitional cell carcinoma is a type of carcinoma that originates in the cells that cover the urinary tract and renal pelvis. Examples include transitional cell carcinoma of the bladder and renal pelvis. Basal cell carcinoma is a type of carcinoma that originates in basal cells, which are the cells in the lower part of the epidermis (the outer layer of skin). Among sarcomas, osteosarcoma originates in bone. Synovial sarcoma is a type of sarcoma that originates in the synovial membrane, which is the lining of joints. Liposarcoma is a type of sarcoma that originates in adipocytes. Ewing's sarcoma is a type of sarcoma that usually originates in bone, but can also originate in other soft tissues. Among leukemias, acute lymphoblastic leukemia (ALL) is a type of leukemia that begins in white blood cells called lymphoblasts. Acute myeloid leukemia (AML) is a type of leukemia that begins in white blood cells called myeloblasts. Chronic lymphocytic leukemia (CLL) is a type of leukemia that begins in white blood cells (lymphocytes). Chronic myeloid leukemia (CML) is a type of leukemia that begins in white blood cells (myeloid cells). Among lymphomas, Hodgkin lymphoma is a type of lymphoma characterized by the presence of a cell type called Reed-Sternberg cells.Non-Hodgkin lymphoma is a type of lymphoma that does not have Reed-Sternberg cells. Examples include diffuse large B-cell lymphoma, follicular lymphoma, and mantle cell lymphoma. Among the group of melanomas, superficial spreading melanoma is the most common type. Nodular melanoma is a type of melanoma characterized by a raised, uneven appearance. Acral lentiginous melanoma is a type of melanoma that occurs on the palms of the hands, soles of the feet, and under the nails. Among the group of brain tumors, astrocytoma is a neoplasm that begins in astrocytes, which are cells that provide structural support to nerve cells in the brain. Glioma is a neoplasm that begins in glial cells, which are cells that provide support to nerve cells in the brain and spinal cord. Meningioma is a neoplasm that begins in the meninges, which are layers of tissue that cover the brain and spinal cord. Medulloblastoma is a neoplasm that begins in the cerebellum. It is important to note that this list is not exhaustive and that other types of malignant tumors exist and are included in the list of cancers that can be effectively treated with the IAC of the present invention.

[0149] In a further preferred embodiment, the disease is a form of carcinoma or neoplasm selected from the group including, in particular, adenocarcinomas such as lung adenocarcinoma, pancreatic adenocarcinoma, prostate adenocarcinoma and colon adenocarcinoma; squamous cell carcinomas such as squamous cell carcinoma of the lung, head and neck squamous cell carcinoma and cervical squamous cell carcinoma; transitional cell carcinomas such as transitional cell carcinoma of the bladder and renal pelvis; and basal cell carcinomas (BCCs) such as nodular BCC, nodular BCC, superficial BCC, pigmented BCC, sclerosing BCC, cystic BCC and invasive BCC. Lung adenocarcinoma is a type of lung cancer that begins in the mucus-producing glands in the lungs. Lung adenocarcinoma is the most common type of lung cancer in nonsmokers and is often diagnosed in its later stages because it does not cause symptoms until it has spread. Prostate adenocarcinoma is a type of cancer that begins in the glands that produce prostatic fluid. Prostate adenocarcinoma is the most common type of prostate cancer and is usually diagnosed in older men. Colon adenocarcinoma is a type of cancer that begins in the glands lining the colon and rectum. Colon adenocarcinoma is the most common type of colorectal cancer and often does not cause symptoms until it has progressed. Squamous cell carcinoma is a type of skin cancer that begins in the flat, flaky cells that make up the outer layer of the skin. There are several types of squamous cell carcinoma, including squamous cell carcinoma of the lung, which is a type of lung cancer that begins in the squamous cells that line the airways. Head and neck squamous cell carcinoma is a type of cancer that begins in the squamous cells that line the mouth, nose, throat, and other parts of the head and neck. Cervical squamous cell carcinoma is a type of cancer that begins in the squamous cells that line the cervix. Transitional cell carcinoma is a type of cancer that begins in cells that can change shape, such as the cells that line the bladder and renal pelvis. Transitional cell carcinoma of the bladder is a type of bladder cancer that begins in the cells that line the bladder. Transitional cell carcinoma of the renal pelvis is a type of cancer that begins in the cells lining the renal pelvis, which is part of the kidney that collects urine.

[0150] Basal cell carcinoma (BCC) is a type of skin cancer that originates in the cells that make up the lower layers of the skin. Several subtypes of BCC exist, including: Nodular BCC, which is usually a raised, solid bump that is pink, red, or flesh-colored; Superficial BCC, a type of BCC that grows slowly and remains near the surface of the skin; Pigmented BCC, a type of BCC that is dark in color, often caused by an increase in the number of pigment-producing cells; Sclerosing BCC, a type of BCC that is hard, scar-like, and white or yellow in color; Cystic BCC, a type of BCC that has a fluid-filled central region; and Invasive BCC, a type of BCC that grows in deeper layers of the skin and is more difficult to treat.

[0151] Notably, all of these cancers or neoplasms can be favorably treated with IAC according to the present invention.

[0152] In a further preferred embodiment, the disease is a neoplasm of a form of sarcoma selected from the group including osteosarcoma, synovial sarcoma, liposarcoma, and Ewing's sarcoma. Osteosarcoma is a type of neoplasm that begins in the cells that make up bone. Osteosarcoma is the most common type of bone cancer and is most commonly seen in teenagers. Synovial sarcoma is a type of neoplasm that begins in the cells that cover the joints. Synovial sarcoma can occur in any part of the body but is most commonly seen in the legs and arms. Liposarcoma is a type of neoplasm that begins in the cells that make up fat. Liposarcoma is the most common type of soft tissue sarcoma and can occur in any part of the body where fat is present. Ewing's sarcoma is a type of neoplasm that begins in the cells of bone or soft tissue. Ewing's sarcoma is most commonly seen in the legs, arms, pelvis, and chest. Notably, all of these neoplasms can be favorably treated with IAC according to the present invention.

[0153] In a further preferred embodiment, the disease is a neoplasm or cancer, particularly a form of leukemia, selected from the group including acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), and chronic myeloid leukemia (CML). It originates in hematopoietic cells in the bone marrow. ALL affects lymphocytes, a type of white blood cell, and typically progresses rapidly if left untreated. Acute myeloid leukemia (AML) is a type of neoplasm that originates in hematopoietic cells in the bone marrow. AML affects myeloid cells that produce red blood cells, white blood cells, and platelets. AML typically progresses more rapidly than other types of leukemia. Chronic lymphocytic leukemia (CLL) is a type of neoplasm that originates in hematopoietic cells in the bone marrow. CLL affects lymphocytes and typically progresses slowly over time. Chronic myeloid leukemia (CML) is a type of neoplasm that originates in hematopoietic cells in the bone marrow. CML affects myeloid cells and typically progresses slowly over time. Notably, all of these neoplasms can be favorably treated with IAC according to the present invention.

[0154] In a further preferred embodiment, the disease is a form of lymphoma cancer selected from the group including Hodgkin lymphomas and non-Hodgkin lymphomas, such as diffuse large B-cell lymphoma, follicular lymphoma, and mantle cell lymphoma. Hodgkin lymphoma is a type of cancer that begins in the lymphatic system, which is part of the body's immune system. Hodgkin lymphoma is characterized by the presence of abnormal cells called Reed-Sternberg cells. Non-Hodgkin lymphoma is a type of cancer that begins in the lymphatic system. Non-Hodgkin lymphoma is a diverse group of cancers that can affect different types of cells in the lymphatic system. Some common subtypes of non-Hodgkin lymphoma include: diffuse large B-cell lymphoma: this is a type of non-Hodgkin lymphoma that affects B cells, a type of white blood cell involved in the immune response; follicular lymphoma: this is a type of non-Hodgkin lymphoma that affects B cells. Follicular lymphoma typically progresses more slowly than other types of non-Hodgkin lymphoma. Mantle cell lymphoma: This is a type of non-Hodgkin lymphoma that affects B cells. Mantle cell lymphoma is typically a rapidly progressing, invasive type of lymphoma; notably, all of these cancers can be favorably treated with IAC according to the present invention.

[0155] In a further preferred embodiment, the disease is a cancer or neoplasm, particularly a form of melanoma, selected from the group including superficial spreading melanoma, nodular melanoma, and acral lentiginous melanoma. Superficial spreading melanoma is a type of skin cancer that originates in the pigment cells (melanin-forming cells) of the skin. Superficial spreading melanoma is the most common type of melanoma and typically spreads horizontally before growing vertically. This type of melanoma often presents as flat or slightly raised lesions with irregular borders and multiple colors. Nodular melanoma is a type of skin cancer that originates in the pigment cells (melanin-forming cells) of the skin. Nodular melanoma is characterized by raised, dome-shaped lesions that are usually black or blue in color. This type of melanoma tends to grow vertically and can rapidly spread to other parts of the body. Acral lentiginous melanoma is a type of skin cancer that originates in the pigment cells (melanin-forming cells) of the skin. This type of melanoma typically occurs on the palms of the hands, soles of the feet, or under the nails. Acral lentiginous melanoma is often misdiagnosed as a bruise or discoloration, making it difficult to detect in its early stages.

[0156] Notably, all of these neoplasms can be favorably treated with IAC according to the present invention.

[0157] In a further preferred embodiment, the disease is a neoplasm of a form of brain tumor, selected from the group including astrocytoma, glioma, meningioma, and medulloblastoma. Astrocytoma is a type of brain tumor originating from astrocytes, which are supporting cells in the brain. Astrocytoma is a type of glioma. Glioma is a type of brain tumor originating from glial cells, which are supporting cells in the brain. Gliomas can range from low-grade (slow-growing) to high-grade (rapid-growing) tumors. Meningioma is a type of brain tumor originating from the meninges, the membranes surrounding the brain and spinal cord. Meningiomas are usually benign (not cancerous) and grow slowly. Medulloblastoma is a type of brain tumor originating from the lower part of the brain (cerebellum). Medulloblastomas grow rapidly and can spread to other parts of the body. Medulloblastoma is most common in children and young adults.

[0158] Notably, all of these cancers can be favorably treated with IAC according to the present invention.

[0159] In a further preferred embodiment, the cancerous disease is a cancer selected from the group including prostate cancer, bladder cancer, neuroendocrine tumors (NETs), melanoma, small cell lung cancer, pancreatic cancer, breast cancer, colorectal cancer, leukemia, liver cancer, lung cancer, ovarian cancer, prostate cancer, gastric cancer, thyroid cancer, and uterine cancer.

[0160] Prostate cancer develops in the prostate gland, a gland in the male reproductive system. Bladder carcinoma forms in the bladder, the organ that stores urine. Neuroendocrine tumors (NETs) are a type of neoplasm that begins in cells that release hormones and other substances into the bloodstream. Melanoma is a type of cutaneous neoplasm that begins in cells that produce pigment in the skin. Small cell lung carcinoma is a type of lung cancer that typically grows and spreads rapidly. Pancreatic carcinoma is a cancer that develops in the pancreas, an organ located behind the stomach that plays a role in digestion and blood sugar control. Breast cancer is a cancer that develops in breast tissue and is most common in women, but can also occur in men. Colorectal cancer is a cancer that begins in the colon or rectum, which are parts of the large intestine. Leukemia is a neoplasm that begins in hematopoietic tissue, such as bone marrow, and causes the production of a large number of abnormal white blood cells. Liver cancer begins in the liver, an organ that filters toxins from the blood and aids in digestion. Lung cancer begins in the lungs and can be classified into small cell lung cancer and non-small cell lung cancer. Ovarian cancer begins in the ovaries, two small organs in the female reproductive system that produce eggs. Stomach cancer begins in the stomach, the organ that helps digest food. Thyroid cancer begins in the thyroid gland, a gland in the cervix that produces hormones that regulate the body's metabolism. Uterine cancer begins in the uterus, the female reproductive organ in which a fetus develops.

[0161] Notably, all of these cancers can be favorably treated with IAC according to the present invention.

[0162] In a further preferred embodiment, the target cellular environment is the tumor immune microenvironment (TIME). As used herein, the term “tumor immune microenvironment (TIME)” preferably refers to the complex interactions between cancer cells, immune cells, and various molecules and structures present in the tissue surrounding the tumor. Tumor-associated immune cells, including T cells, B cells, natural killer cells, and dendritic cells, can infiltrate the tumor microenvironment and play a role in cancer immunity. There are different types of immune cells that can be found in the tumor microenvironment, including tumor-infiltrating lymphocytes (TILs), which are immune cells that have infiltrated the tumor; tumor-associated macrophages (TAMs), which are a type of leukocyte that can promote tumor growth and spread; myeloid-derived suppressor cells (MDSCs), which can suppress the immune response against cancer; regulatory T cells (Tregs), which can suppress the immune response against cancer; and / or dendritic cells, which play a key role in activating the immune response against cancer. TIME also includes the presence of molecules such as TGF-β, IL-10, and VEGF, which can promote tumor growth and inhibit the immune response. The balance of these different types of cells and molecules in the tumor microenvironment can influence cancer progression and the effectiveness of cancer treatment. Understanding the tumor immune microenvironment is crucial for developing new cancer therapies that target the interactions between cancer cells and the immune system.

[0163] When targeting target cells during time-induced angiogenesis, the IAC of the present invention can favorably inhibit angiogenesis. Furthermore, the IAC of the present invention can eliminate the tumor base and favorably remove apoptotic products.

[0164] In a further preferred embodiment, the target cells are preferably tumor immunomicroenvironment (TIME) target cells selected from the group including tumor-infiltrating lymphocytes (TILs), tumor-associated macrophages (TAMs), TIME-associated myeloid-derived suppressor cells (MDSCs), TIME-associated regulatory T cells (Tregs), and TIME-associated dendritic cells.

[0165] In this regard, those skilled in the art will immediately recognize and understand that TIME, and the cells within TIME, respectively, represent and / or are typical of TIME, and / or characteristic of TIME. However, there are other cell types that may be present in other healthy tissues as well. For example, fibroblasts, while certainly prominent in TIME, are present in many other tissues within the body of the subject. In particular, fibroblasts may have different roles and / or characteristics in tumor tissue and TIME, respectively. Tumor-infiltrating fibroblasts in the context of this invention may refer specifically to cancer-associated fibroblasts, but in some embodiments, inflammatory fibroblasts may also be included.

[0166] In a further preferred embodiment, the presence and / or overexpression of a target structure indicates the cancerous disease in question, and the target structure is selected from the group including growth factor receptors, antigenic alterations, cell adhesion molecules, angiogenic factors, proteases, extracellular matrix molecules, and cell surface molecules.

[0167] It should be understood that the more specific a target structure is to the presence of cancer, particularly malignant cancer cells, the more specifically this IAC approach can target such target cells and / or the target cell environment. For example, overexpression of the HER2 / neu gene is characteristic of certain types of breast cancer. The presence of this overexpression can be used to help determine the best treatment strategy for a patient / animal.

[0168] Overexpression of a particular target structure in cells means that an increased amount of that structure is present compared to normal, non-malignant cells. In relation to cancer, overexpression of a particular gene or protein can indicate cancer. In cancer, gene mutations and other changes can lead to unregulated proliferation and division of cells. Some of these changes can lead to the overproduction of certain genes or proteins that promote cancer growth. When these genes or proteins are present at abnormally high levels, they are said to be overexpressed. Overexpression of a particular target structure in cancer cells is used as a marker to help diagnose and classify different types of cancer. In this IAC therapeutic approach, overexpression would result in a statistically increased probability that the IAC will bind to cancer cells. Where the “presence” of a target structure is referred to herein as indicating the cancer in question, this preferably includes the fact that normal, non-malignant cells do not exhibit such presence of the target structure. Therefore, treatment with IAC is specific to the target structure and does not involve site-effects for nonspecific binding or binding to non-target cells, or preferably only involves limited site-effects for nonspecific binding or binding to non-target cells.

[0169] A group of target structures, including growth factor receptors, antigenic alterations, cell adhesion molecules, angiogenic factors, proteases, extracellular matrix molecules, and cell surface molecules, play key roles in the development and progression of cancer, and therefore can serve as excellent indicators of cancer. Growth factor receptors are proteins found on the surface of cells that help regulate cell proliferation, division, and survival. In cancer, mutations or increased activity of these receptors can lead to unregulated cell proliferation and division. Antigenic alterations refer to changes in the antigenic properties of cancer cells, making them different from normal cells. These alterations can be used as markers to help diagnose cancer and monitor its progression. Cell adhesion molecules are proteins that help cells stick together and form tissues. In cancer, changes in these molecules can lead to the detachment of cancer cells from the primary tumor and their spread (metastasis) to other parts of the body. Angiogenic factors are proteins that promote the growth of new blood vessels. In cancer, increased production of angiogenic factors can help supply oxygen and nutrients to growing tumors. Proteases are enzymes that break down proteins. In cancer, several proteases can contribute to the degradation of the extracellular matrix and the disruption of normal tissue structure, enabling cancer cells to invade and spread. The extracellular matrix is ​​a network of proteins and carbohydrates that provides structural support to tissues. In cancer, changes in the composition of the extracellular matrix can contribute to the invasion and spread of cancer cells.

[0170] Cell surface molecules are proteins found on the surface of cells that help regulate intercellular interactions and information exchange. In cancer, changes in these molecules may contribute to the abnormal behavior of cancer cells and their ability to evade the immune system. Therefore, these target structures are also potential markers that help diagnose and classify different types of cancer, and their measurement may be useful in monitoring the response to treatment and predicting prognosis. Accordingly, these structures are preferred target structures for selecting the target structure binding moiety of the IAC of the present invention, especially when the presence and / or overexpression of the target structure indicates the cancerous disease of the subject. It is particularly preferable that the target structure is expressed by the target cell. Thereafter, preferably, the target structure is a cell surface protein of the target cell. It is even more preferable that the target structure is exposed on the cell surface of the target cell. This allows for a higher probability of binding of the target structure binding moiety to the target structure on the target cell. In this specification, it is particularly conceivable that the target structure is preferably a cell surface receptor molecule of the target cell.

[0171] Cell surface receptor molecules are located on the cell surface and become reachable for therapeutic intervention of IACs and their target structure binding moieties; therefore, they are particularly preferred as target structures for IACs in the present invention.

[0172] In further preferred embodiments, the presence and / or overexpression of an immunoinducing compound (IAC) target structure according to the present invention indicates the target cancer disease, the target structure indicates the presence of TIME, and is preferably selected from the group comprising vascular endothelial growth factor (VEGF), matrix metalloproteinase (MMP), tumor necrosis factor-α (TNF-α), interleukin (IL), epidermal growth factor (EGF), transforming growth factor-β (TGF-β), insulin-like growth factor (IGF), platelet-derived growth factor (PDGF), and cytokines.

[0173] In a further preferred embodiment, the presence and / or overexpression of a target structure indicates the target cancer disease, and the target structure is a fibroblast-activating protein (FAP).

[0174] FAP, as used herein, is also referred to as “fibroblast-activating protein” and is used interchangeably with “FAP” or “fibroblast-associated protein.” Fibroblast-activating protein (FAP) is a cell surface protein expressed on fibroblasts, cells that play a key role in the formation and maintenance of connective tissue and are involved in wound healing and tissue remodeling. FAP has been found to be involved in several biological processes, including tumor growth and angiogenesis. FAP is also used as a biomarker to identify fibroblasts in various tissues. FAP was first described by Wolfgang J. Rettig et al. (Wolfgang J. Rettig, P. Garin-Chesa and Lloyd J. Old (MSKCC) 1990). FAP is highly expressed in CAFs in 90% of all cancer types. Cancer-associated fibroblasts (CAFs) are extremely widespread in the tumor microenvironment (TME) of many cancer entities, and CAFs persist in the TME throughout all malignant stages of tumorigenesis. This revealed that increased FAP expression is present on CAFs in the tumor-associated stroma of more than 30 different malignant tumors, as well as on the cell surfaces of certain cancer types (e.g., PDAC cells, gastric cancer cells, ovarian cancer, breast cancer, sarcoma, etc.). This indicates that FAP expression in normal tissues is typically very low, or even undetectable by standard methods known in the art. CAFs express FAP, and therefore FAP is a pan-tumor target and, consequently, a preferred target structure in the IAC embodiments of the present invention. Since FAP is not expressed or is only mildly expressed on normal fibroblasts, FAP expression in normal tissues is extremely low (Hanahan et al. Cell, 2011). The inventors also recognize that FAP expression is often associated with poorer clinical outcomes and tumor progression. However, while not wishing to be bound by this theory, there are controversial findings in certain cancer types, and the precise mechanisms of action of FAP on tumor growth, migration, and invasion are not yet fully understood. Furthermore, the tumor-suppressing effect of FAP was also described.Accordingly, FAP-2286 and 3BP-3940 are examples of optimized FAP-binding peptide conjugates linked to DOTA (chelating agents) for post-traumatic imaging therapy (PTRT) applications. Thus, these conjugates may also be possible target structure binding moieties in embodiments of the IAC of the present invention.

[0175] In further preferred embodiments, the target structure is selected from tumor-associated antigens (TAAs), tumor-specific antigens, and tumor germline antigens, preferably carcinoembryonic antigen (CEA), carcinometris antigen (CTA), mucin-1 (MUC1), Her2 / neu (also known as human epidermal growth factor receptor 2), alpha-fetoprotein (AFP), tyrosinase, cancer / testis antigen (CTA), epidermal growth factor receptor (EGFR), and prostate-specific antigen (PSA), GD2 The group includes (ganglioside-2), MART-1 / Melan-A, sialyl Lewis X (sLeX), NY-ESO-1 (New York esophageal squamous cell carcinoma-1), cyclin B1, p53 tumor antigen, cancer germline antigen (CGA), MAGE (melanoma-associated antigen), and PSMA (prostate-specific membrane antigen), somatostatin receptor (SSTR), TME-specific receptor, fibroblast-activating protein (FAP), and kallikrein-4 (KLK4).

[0176] CEA (carcinoembryonic antigen) is a protein typically found in the tissues of developing fetuses and in small amounts in the blood of healthy adults. Elevated levels of CEA in the blood may indicate the presence of certain cancers, such as colorectal cancer or pancreatic cancer. CTA (cancotesticular antigen) is a protein typically expressed only in the testes, but can also be expressed in certain cancers, including germ cell tumors and lung cancer. CTA is considered a promising target for cancer immunotherapy. MUC1 (mucin-1) is a protein expressed on the surface of many types of cancer cells, including breast, ovarian, and pancreatic cancers. MUC1 is often overexpressed in cancer and may be a useful target for diagnosis and treatment. Her2 / neu is a type of cell surface receptor involved in cell proliferation and division. Overexpression of Her2 / neu is associated with high-grade morphologies of breast cancer and may be a target for therapies such as Herceptin. AFP (alpha-fetoprotein) is a protein produced by developing fetuses and certain cancers, including liver and testicular cancers. Elevated levels of AFP in the blood can be a sign of cancer. Tyrosinase is an enzyme involved in the production of melanin, the pigment that gives color to the skin, hair, and eyes. Tyrosinase is overexpressed in melanoma, a type of skin cancer. EGFR (epidermal growth factor receptor) is a type of cell surface receptor involved in cell proliferation and division. Overexpression of EGFR is associated with certain cancers, including lung cancer, breast cancer, and ovarian cancer. PSA (prostate-specific antigen) is a protein produced by the prostate gland. Elevated levels of PSA in the blood can be a sign of prostate cancer. GD2 (ganglioside-2) is a carbohydrate molecule expressed on the surface of certain cancer cells, including neuroblastoma, a type of childhood cancer. GD2 is a promising target for cancer immunotherapy. MART-1 / Melan-A is a protein expressed on the surface of melanoma cells, a type of skin cancer. MART-1 / Melan-A is a target for cancer immunotherapy. sLeX (sialyl Lewis X) is a carbohydrate molecule expressed on the surface of certain cancer cells, including breast and colon cancer cells. sLeX is a target of cancer immunotherapy. NY-ESO-1 (New York esophageal squamous cell carcinoma-1) is a carcinogenic antigen expressed in various cancers, including esophageal cancer, lung cancer, and ovarian cancer.NY-ESO-1 is a promising target for cancer immunotherapy. Cyclin B1 is a protein involved in regulating cell division. Overexpression of cyclin B1 is associated with certain cancers, including breast and ovarian cancer. The p53 tumor antigen is a protein involved in regulating the cell cycle and preventing tumor formation. Mutations in the p53 gene are common in many types of cancer, including lung, breast, and colorectal cancer. CGA (oncogerm cell lineage antigen) is a protein that is normally expressed only in germ cells, but can also be expressed in certain cancers, including testicular and ovarian cancer. CGA is considered a promising target for cancer immunotherapy. MAGE (melanoma-associated antigen) is an oncogerm cell lineage antigen that has been found to be expressed in various types of cancer, including melanoma, lung, and testicular cancer. The MAGE gene family is considered a promising target for cancer immunotherapy due to its limited expression in normal tissues. Prostate-specific membrane antigen (PSMA) is a cell surface receptor highly expressed in prostate cancer and is used as a biomarker for the diagnosis and treatment of prostate cancer. Somatostatin receptor (SSTR) is a cell surface receptor widely expressed in various types of cancer, including neuroendocrine tumors, and is used as a target for radionuclide therapy. TME-specific receptors refer to specific receptors expressed in the tumor microenvironment and used as targets for cancer immunotherapy. Fibroblast-activating protein (FAP) is a cell surface protein highly expressed in various types of cancer, including colorectal cancer, and is considered a promising target for cancer immunotherapy. Kallikrein-4 (also known as KLK4) is a member of the human kallikrein-associated peptidase (KLK) family of serine proteases. KLK4 is involved in various physiological processes, including skin desquamation, hair growth, and blood pressure regulation. KLK4 is also linked to the development and progression of various diseases, including cancer. Studies have shown that KLK4 expression is elevated in several types of cancer and is associated with poor prognosis and tumor progression. KLK4 has been proposed as a potential therapeutic target and a biomarker for cancer diagnosis and prognosis.

[0177] Therefore, all of the aforementioned target structures are advantageous target structures for selecting the target structure binding portion of the IAC of the present invention that binds to the aforementioned target structures, and the presence and / or overexpression of the target structures indicates the cancerous disease of the subject.

[0178] In a further preferred embodiment, the target structure is a tumor immune microenvironment (TIME)-associated antigen.

[0179] In a further preferred embodiment, the target structure is a tumor-related biomarker. As used herein, the term “tumor-related biomarker” preferably refers to a specific molecule, such as a protein or genetic marker, that is detectable in a biological sample and indicates the presence, progression, or characterization of a tumor. These biomarkers serve as indicators of tumorigenesis and are beneficial in the field of oncology for diagnostic, prognostic, or therapeutic purposes. Therefore, using tumor-related biomarkers as target structures is particularly advantageous because tumor-related biomarkers are usually well-characterized and understood, and can facilitate early detection, accurate diagnosis, and targeted control of progression for treatment.

[0180] In a further preferred embodiment, the target structure is a disease-related biomarker indicating the presence of TIME, preferably FAP. Targeting a disease-related biomarker in the tumor microenvironment (TIME), as exemplified by FAP, is particularly advantageous because it provides a specific and reliable approach for immunomodulation and enables precise intervention by the IAC of the present invention tailored to the presence of a pathological condition.

[0181] In a further preferred embodiment, the target structure is a disease-related biomarker, and the presence and / or overexpression of the target structure indicates the cancerous disease in question.

[0182] In a further preferred embodiment, the target structure is a peptide or protein fragment.

[0183] In a further preferred embodiment, the target structure binding moiety can bind to an extracellular target structure, and the presence and / or overexpression of the target structure indicates the cancerous disease of the subject.

[0184] In a further preferred embodiment, the target cells are tumor cells, and / or the target cell environment is the tumor cell environment, in particular the tumor immunomicroenvironment (TIME).

[0185] As used herein, the term “target cell environment” preferably means the tumor cell environment, in particular the tumor immune microenvironment (TIME).

[0186] In a further preferred embodiment, the target structure-binding moiety is a biomarker having specificity for binding to the target structure, and the presence and / or overexpression of the target structure indicates the target cancer disease.

[0187] As used herein, the term “biomarker” preferably refers to a biological molecule or feature that can be measured in a sample from a patient / animal (such as blood or tissue) and used as an indicator of a biological or medical condition, such as the presence or absence of a disease, particularly a cancerous disease according to the present invention, or a response to treatment. Biomarkers are often used as diagnostic or prognostic tools and to monitor progression or response to treatment. In a patent context, a biomarker may refer to a specific protein, gene, or other biological feature specifically claimed as a diagnostic tool, or more generally, a biomarker may be used to describe a method for measuring a biological feature to diagnose or monitor a disease. In the context of the present invention, known biomarkers indicating a cancerous disease in question are considered to be advantageously usable as target structure binding sites.

[0188] In a further preferred embodiment, the target structure binding portion is a ligand molecule capable of binding to the target structure.

[0189] As used herein, the term “ligand” preferably refers to the term as used in the immunological context in the art, particularly when it means that “the target structure binding portion is a ligand.” Thereafter, those skilled in the art will recognize that, unlike its use in other contexts of the invention which may represent a radioactivity-capturing portion that essentially acts as a cage for radioactive metals, the term “ligand” in its immunological context primarily refers to a molecule involved in binding to a target structure. Preferably, for immunological purposes, a ligand is defined as a molecule involved in immune interactions.

[0190] Therefore, in a preferred embodiment, the target structure binding moiety is a ligand molecule selected from the group including molecules that bind SSTR2, PSMA, CXCR4, Her2-neu, and FAP, preferably selected from the group including FAP-2286, FAP-46, and 3BP-3940.

[0191] FAP-2286 has been extensively studied in preclinical evaluations for targeted radionuclide imaging and therapy, and is therefore a particularly preferred FAP-binding ligand for use as the target structure binding moiety of the IAC of the present invention (Zboralski et al., European Journal of Nuclear Medicine and Molecular Imaging (2022) 49:3651-3667, https: / / doi.org / 10.1007 / s00259-022-05842-5).

[0192] In a further preferred embodiment, the target structure binding moiety is preferably a molecule selected from structures targeting SSTR2, PSMA, FAP, CXCR4, Her2-neu, or any other ligand that specifically targets any of these receptors, preferably including radiolabeled analogs, particularly synthetic or biological molecules or structures.

[0193] In a further preferred embodiment, the target structure-binding moiety is selected from the group comprising TIME-binding molecules, preferably avidin. Those skilled in the art will then know that avidin binds to TIME in a nonspecific manner. While not bound by theory, the inventors hypothesize that the nonspecific binding of avidin to TIME is due to the acidic environment in TIME, and therefore the binding may be pH-dependent. At the same time, there is evidence that in some cancer types, avidin binds directly in the tumor region, such as in bladder carcinoma.

[0194] In a further preferred embodiment, the target structure binding moiety can bind to a target structure in the target tumor immune microenvironment (TIME), preferably the target structure is selected from bladder carcinoma-related or specific target structures. Such bladder carcinoma-specific target structures are known in the art and have been reported, for example, particularly by Paganelli et al.

[0195] In a further preferred embodiment, the target structure binding portion can bind to a target structure on a target tumor cell, and the target structure is selected from the target structures described in any of the preceding claims.

[0196] In a further preferred embodiment, the target structure binding moiety can bind to at least one, preferably one or more, and particularly at least two target structures in the tumor cell or tumor immune microenvironment (TIME), the target structures being selected from the target structures described in any of the preceding claims. Thus, the IAC of the present invention can be construed such that the target structure binding moiety can bind to at least one, preferably one or more, and particularly at least two target structures in the tumor cell or tumor immune microenvironment (TIME), and / or the IAC of the present invention may comprise one or more, preferably one or more, more preferably one or more, and particularly at least two target binding moieties that can bind to at least one, preferably one or more, and particularly at least two target structures in each of the tumor cells of the subject.

[0197] In a further preferred embodiment of the immunoinducing compound (IAC) according to the present invention, the IAC comprises at least two, preferably more than two, target structure binding moieties (2), wherein the first target structure binding moiety (2) is the same as or different from the second target structure binding moiety (2).

[0198] In a further preferred embodiment of the immunoinducing compound (IAC) according to the present invention, the target structure binding moiety (2) of the immunoinducing compound (IAC) comprises an affilin (21).

[0199] As used herein, the term “affilin” preferably refers to a type of small protein or peptide engineered for therapeutic and diagnostic applications. Affilins are preferably derived from a natural human protein backbone known as “Stephin A.” Affilins are preferably designed using a specific molecular evolution technique called Affimer technology, which is known to those skilled in the art and was first developed by Avacta Life Sciences. Affilins are typically characterized by their small size, stability, and ability to bind to specific target molecules with high affinity. Due to their compact size and robust nature, affilins offer advantages such as rapid tissue penetration and efficient production. Therefore, affilins can be advantageously included in immunoinducing compounds according to the present invention. Generally, those skilled in the art are familiar with affilins, and in particular, can develop affilins directed to specific target structures, as disclosed, for example, in Lorey et al. (Lorey S, Fiedler E, Kunart A, Nerkamp J, Lange C, Fiedler M, Bosse-Doenecke E, Meysing M, Gloser M, Rundfeldt C, Rauchhaus U, Hanssgen I, Gottler T, Steuernagel A, Fiedler U, Haupts U. Novel ubiquitin-derived high affinity binding proteins with tumor targeting properties. J Biol Chem. 2014 Mar 21;289(12):8493-507. doi:10.1074 / jbc.M113.519884. Epub 2014 Jan 28. PMID:24474690; PMCID:PMC3961674). As discussed herein, “Affilin” refers to a class of high-affinity and specific binding molecules developed for the treatment and diagnosis of cancer. These molecules are derived from a dimeric ubiquitin library and are selected for fibronectin extradomain B (ED-B), a target primarily expressed in tumor tissue.Affilin molecules are scaffold-based binding proteins possessing outstanding biophysical and biochemical properties, including high thermal and serum stability, as well as strong in vitro target binding and in vivo tumor accumulation. The ultimate goal in developing affilin is to target effector molecules, such as toxins, cytokines, or radiolabeled molecules, to tumor cells, and to bind to target structures as target structure binding moieties according to the present invention, as particularly discussed herein. These molecules are designed to possess optimal properties for cancer treatment, including high tumor accumulation while maintaining low levels in healthy tissues and blood, which makes these molecules particularly interesting as target structure binding molecules within the scope of the present invention. Affilin are characterized by short systemic circulation, making them highly suitable for combination with effector molecules and half-life extension technologies, such as HEAD linkers.

[0200] In particular, affilin can be linked to or attached to the immune-inducing portion.

[0201] Furthermore, affilin may be bound to or attached to chelating agents. In particular, affilin may be bound to and / or to immune-inducing moieties.

[0202] In a further preferred embodiment, the immune cells are selected from the group including T cells, B cells, natural killer (NK) cells, macrophages, dendritic cells, monocytes, neutrophils, mast cells, and eosinophils.

[0203] In a further preferred embodiment, the immune cell includes an immune cell receptor to which an immune-attracting moiety can be bound.

[0204] In a further preferred embodiment, immune cells are attracted to and / or to the target cell environment.

[0205] In a further preferred embodiment, the target structure binding portion and the immune-inducing portion are linked by a linker portion.

[0206] As used herein, the term “linker” preferably refers to any structure that covalently links different parts of the IAC. Thereafter, more preferably, the covalent bond itself may represent the simplest form of a linker in the context of this application. In particular, the target structure binding portion (2) and the immune attracting portion (3) are linked by a linker portion (8). The linker portion contains or consists of covalent bonds. It should therefore be understood that the linker portion contains or consists of covalent bonds, and these covalent bonds connect the immune attracting portion (3) and the structure binding portion (2) to any other part of the IAC. In a further preferred embodiment, the linker portion contains or consists of a hydrocarbon portion.

[0207] In further preferred embodiments, each linker moiety may be individually selected from the group including amide-, carboxylic acid amide-, phosphinate-, alkyl-, triazole-, thiourea-, ethylene-, maleimide residues, -(CH2)m-, -(CH2CH2O)m- and (CH2)mNH-m, or others.

[0208] In a further preferred embodiment, both the target structure binding portion (2) and the immune inducer portion (3) are connected using a linker portion (8). Thus, in a further preferred embodiment, the connection between the target structure binding portion (2) and the immune inducer portion (3) via the linker portion (8) can take various forms. This includes the possibility of using two separate linkers or the same molecule to adapt the linear structure of several identical or different linkers, or the possibility of taking a Y-shaped structure and potentially incorporating two or more linkers. These linkers may be identical or different, and it is essential to recognize that this allows flexibility in the design of the immune inducer compound (IAC) to optimize the structural and functional characteristics of the IAC. Perhaps both the target structure binding portion (2) and the immune inducer portion (3) are connected to each other using the same or different linker portions (8).

[0209] In a further preferred embodiment, the linker portion includes a spacer.

[0210] Such spacers can advantageously allow for a distance between the target structure binding site and at least one immune-attracting site, thereby reducing steric collisions. Such spacers may be particularly advantageous when more than one immune-attracting site is linked to the target structure binding site, or when one or both of the immune-attracting site and / or target structure binding site have relatively small molecular sizes.

[0211] This may be particularly important in embodiments where the target structure binding moiety is avidin. Here, the spacer and the length of the spacer can be advantageously selected. Particularly preferably, when avidin is the target structure binding moiety, the spacer has a length of 22.2 angstroms and / or a chain length of 18 atoms.

[0212] In a further preferred embodiment, both the target structure binding portion (2) and the immune induction portion (3) are connected using one or more linker portions (8), where the first linker portion is connected to the second linker portion.

[0213] In a further preferred embodiment, both the target structure binding portion (2) and the immune induction portion (3) are connected using a linker portion (8).

[0214] In a further preferred embodiment, the IAC comprises a chelating agent, the chelating agent being linked to a target structure binding moiety (2) and / or an immune-inducing moiety (3).

[0215] It is important to understand that chelates form complexes with metal ions, including radionuclides, via multiple coordination bonds (donation bonds). The term "chelating agent" describes the way in which a chelating agent "grasps" a metal ion via multiple coordination sides. In the case of radionuclides, the chelating agent forms a robust structure around the metal ion, producing a stable complex. An advantage is that this chelate can be covalently bonded to any other chemical structure, which is not applicable to free radioactive metals. Furthermore, by selecting the appropriate chelate for the appropriate metal, high stability can be given to the entire complex. This stability can prevent the release of radionuclides into the environment, making it easier to handle and transport radionuclides in IACs of the present invention to target cells and / or target cell environments.

[0216] In a further preferred embodiment, the linker portion (8) comprises a chelating agent. In this regard, the chelating agent may be linked to the target structure binding portion (2) and / or the immune-inducing portion (3) by one or more linker portions, each linker portion being independently selected and may be the same as or different from one another.

[0217] In a further preferred embodiment, the IAC comprises a chelating agent, the chelating agent being linked to a target structure binding moiety (2) and / or an immune-inducing moiety (3).

[0218] In a further preferred embodiment, the chelate can bind a radioactive moiety, preferably a radionuclide.

[0219] The incorporation of a chelating agent into the immunoinducing compound of the present invention is particularly advantageous because it enhances its versatility and enables targeted binding to specific structures and efficient coordination of metal ions, especially radionuclides, for desired therapeutic purposes.

[0220] In further preferred embodiments, the chelating agent is EDTA (ethylenediamine tetraacetate), EDTMP (diethylenetriamine penta(methylenephosphonic acid)), DTPA (diethylenetriamine pentaacetate) and its derivatives, DOTA (dodeca-1,4,7,10-tetraamine tetraacetate), DOTAGA (2-(1,4,7,10-tetraazacyclododecane-4,7,10)-pentanedioic acid) and other DOTA derivatives, TRITA (trideca-1,4,7,10-tetraamine tetraacetate) TETA (tetradeca-1,4,8,11-tetraamine tetraacetate) and its derivatives, NOTA (nona-1,4,7-triamine triacetate) and NOTAGA (1,4,7-triazacyclononane, 1-glutaric acid, 4,7-acetate) and its derivatives, NOPO (1,4,7-triazacyclononane-1,4-bis[methylene(hydroxymethyl)phosphonic acid]-7-[methylene(2-carboxyethyl)phosphonic acid]), PEPA (pentadeca-1,4,7,10,13-pentamine tetraacetate) Acetate), HEHA (Hexadeca-1,4,7,10,13,16-Hexaaminetetraacetate) and its derivatives, HBED (Hydroxybenzylethylenediamine) and its derivatives, DEDPA and its derivatives such as H2DEDPA (1,2-[[6-(carboxylate-)pyridine-2-yl]methylamino]ethane), DFO (Deferoxamine) and its derivatives, Trishydroxypyridinone (THP) and its derivatives such as YM103, TRAP (Triazacyclononane-phosphonic acid), TEAP (Te The group is selected from the following: traazabicyclododecane-phosphonic acid) and its derivatives, derivatives such as AAZTA (6-amino-6-methylperhydro-1,4-diazepine-N,N,N',N'-tetraacetate) and DATA ((6-pentanoic acid)-6-(amino)methyl-1,4-diazepine triacetate); SarAr (1-N-(4-aminobenzyl)-3,6,10,13,16,19-hexazabicyclo[6.6.6]eicosane-1,8-diamine) and its salts, aminothiols and their derivatives. Exemplary structures of chelating agents are shown in Figure 2.Those skilled in the art will immediately recognize that such chelating agents can complex with any metal, in particular Ga-68, Lu-177, Ac-225, or any other radioactive metal of interest.

[0221] In a further preferred embodiment, the chelating agent can bind to a radioactive moiety (9), preferably the radioactive moiety (9) is a radionuclide (9).

[0222] In a further preferred embodiment, the chelating agent is bound to the radioactive moiety (9), and preferably the radioactive moiety (9) is a radionuclide (9).

[0223] In a further preferred embodiment, the radioactive portion (9) is a radionuclide (9) selected from the group consisting of gamma emitters, positron emitters, Auger emitters, beta emitters, and alpha emitters. Thereafter, those skilled in the art will recognize that therapeutic and diagnostic nuclides can encompass a variety of radiation types, including alpha, beta, and gamma emitters.

[0224] In nuclear medicine, radionuclides are used to generate gamma rays and / or positron emissions for imaging purposes. Gamma rays are high-energy electromagnetic radiation emitted from the nucleus of a radionuclide. Gamma rays can penetrate tissue and are detected by specialized cameras, such as gamma cameras or single-photon emission computed tomography (SPECT) cameras, to generate images of the distribution of radionuclides within the body. Positron emission is the emission of positively charged particles called positrons from the nucleus of a radionuclide. When a positron collides with an electron in the body, the two annihilate each other, producing two gamma rays that travel in opposite directions. These gamma rays can be detected by specialized cameras, such as positron emission tomography (PET) cameras, to generate images of the distribution of radionuclides within the body. Both gamma rays and positron emissions are used for imaging purposes in nuclear medicine, and the choice of radiation depends on the specific imaging requirements. In nuclear medicine, radionuclides are used for both imaging and therapeutic purposes. For therapeutic purposes, beta-emitters and alpha-emitters are commonly used. Beta-emitters are radionuclides that emit beta particles, which are high-energy electrons. These beta particles can damage or kill nearby cells, making them useful for targeted radiotherapy. Some common beta-emitters used for therapeutic purposes include yttrium-90 (Y-90), iodine-131 (I-131), lutetium-177 (Lu-177), and strontium-89 (Sr-89). Alpha-emitters are radionuclides that emit alpha particles, which are heavy, highly charged particles. Alpha particles have a short range and high energy, making them useful for targeted radiotherapy. Some common alpha-emitters used for therapeutic purposes include actinium-225 (Ac-225), lead-212 (Pb-212), and bismuth-213 (Bi-213). In both beta and alpha-emitting therapies, radionuclides are targeted to specific sites within the body, allowing them to deliver high doses of radiation to the targeted area while minimizing exposure to surrounding healthy tissue. This targeted approach makes radionuclides an effective tool for cancer treatment.

[0225] In a further preferred embodiment, the radioactive portion (9) is scandium-44 (44Sc), scandium-47 (47Sc), cobalt-55 (55Co), copper-62 (62Cu), copper-64 (64Cu), copper-67 (67Cu), gallium-66 (66Ga), gallium-67 (67Ga), gallium-68 (68Ga), zirconium-89 (89Zr), yttrium-86 (86Y), yttrium-90 (90Y), niobium-90 (90Nb), ​​techneum-99m (99mTc), indium-111 (111In), samarium-135 (135Sm), praseodymium-140 (140Pr), gadolinium-159 (159Gd), terbius Mu-149 (149Tb), Terbium-160 (160Tb), Terbium-161 (161Tb), Erbium-165 (165Er), Dysprosium-166 (166Dy), Holmium-166 (166Ho), Ytterbium-175 (175Yb), Lutetium-177 (177Lu), Rhenium-186 (186 It is a radionuclide (9) selected from the group including Re, rhenium-188 (188Re), lead-203 (203Pb), lead-212 (212Pb), bismuth-213 (213Bi), actinium-225 (225Ac), fluorine-18 (F-18), iodine-131 (I-131), or astatine-211 (At-211).

[0226] All of these radionuclides are particularly useful when included in the IAC of the present invention for either diagnostic or therapeutic purposes in the diagnosis and treatment of the target cancer.

[0227] In a further preferred embodiment, the chelating agent is selected from the group including acyclic chelating agents, macrocyclic chelating agents, or any other.

[0228] In further preferred embodiments, the chelating agent is selected from the group comprising DOTA, TRITA, TETA, NOTA, PEPA, HEHA, DOTAGA, AAZTA, DATA, EDTA, DTPA, EDTMP, DFO-B, TRAP, DEDPA, H2DEDPA, CP256, YM103, stabilized derivatives of DTPA, and derivatives thereof.

[0229] Linkers may also include half-life extension domains (HEADs). Such domains are known in the art and are interpreted to be able to increase the half-life of molecules in the blood. Several such domains known in the art exist, for example, including albumin-binding moieties, which thus result in binding to blood cells. The use of HEADs may be particularly advantageous when affilin is contained in IACs. Affilin is generally known to have a short residence time in the blood, and therefore conjugation or linkage to HEADs can favorably increase the residence time from a few hours to several days.

[0230] The IAC according to the present invention may include an affilin linked to and / or attached to the half-life extension domain. Thereafter, a chelating agent is linked to and / or attached to the half-life extension domain, and / or an immune-inducing moiety is linked to and / or attached to the half-life extension domain, either additionally or alternatively.

[0231] The basic chemical structural formulas of the above chelates can preferably be obtained from Figures 2A and 2B.

[0232] In a further preferred embodiment, the subject is a mammal, preferably a human.

[0233] According to the present invention and various embodiments thereof, the immunoinducing compounds (IACs) of the present invention suitable for treating target cancer diseases are: - At least one target structure binding moiety (TSM) (2) and at least one immune attraction moiety (IAM) (3) Includes.

[0234] As a result, one or more target structure binding sites and one or more immune-inducing sites, as well as any linker, chelating agent, or affilin, if present, for example in the form of a HEAD, can be arranged in various ways, all of which are considered to be within the scope of the present invention. Several concepts are illustrated in Figure 3.

[0235] For example, in its simplest form, as shown in Figure 3A, one IAM(3) may exist and be linked to one TSM(2) by a covalent linker(8). In another example, the IAM(1) contains a chelating agent(9). For example, as shown in Figures 3B, 3C, or 3D respectively, the IAC(1) includes two linker portions, a first linker(8a) and a second linker(8b). The IAM(3) may be linked to a chelating agent(91) capable of complexing a radionuclide(9) via the first linker(8a), and the TSM(2) may be linked to the chelating agent(91) via the second linker(8b). Alternatively, the chelating agent(91) may be linked to the IAM(3) via the first linker(8a), and the TSM(2) may be linked to the IAM(3) via the second linker(8b). Alternatively, the chelating agent (91) is linked to TSM(2) via a first linker (8a), and IAM(3) is linked to TSM(2) via a second linker (8b).

[0236] As shown in Figure 3F, the linker structure may also include a first linker connected to a branched linker and / or a second linker, and / or a second linker connected to a first and / or third linker. In the embodiments shown, a three-arm linker is shown connecting IAM(3) to TSM(2) and chelating agent (91). In a particularly preferred embodiment, IAC(1) according to the present invention includes one or more TSM(2) and / or one or more chelating agents (91). As shown in Figure 3F, IAM(3) may be connected to TSM(2) and / or chelating agent (91) via a first linker (8a), or vice versa. In particular, IAC(1) according to the present invention may include IAM(3) connected to TSM(2) via a first linker (8a), and TSM(2) connected to chelating agent (91) via a second linker (8b). More specifically, the IAC(1) according to the present invention may include a first IAM(3) connected to a first TSM(2) via a first linker(8a), the TSM(2) being connected to a first chelating agent(91) via a second linker(8b), and the IAC(1) according to the present invention may include a second IAM(3) connected to a second TSM(2) via a further linker, the second TSM(2) being connected to a second chelating agent(91) via yet another linker. Each of the linkers may be the same as or different from one another.

[0237] In certain preferred embodiments, the IAC(1) may include an affilin(21). The affilin(21) may be advantageously linked to and / or attached to a linker(8), the linker comprising a half-life extension domain (HEAD)(11). In particular, a chelating agent(91) may also be linked to and / or attached to the linker(8) comprising the half-life extension domain HEAD(11).

[0238] In further embodiments, the immune-inducing portion (3) is linked to and / or attached to a linker (8) containing a half-life extension domain (HEAD) (11).

[0239] Figures 3G, 3H, 3J, and 3K show several exemplary embodiments of IAC(1) which may include an affilin (21) and a linker (8) having HEAD (11).

[0240] In particular, Figure 3G illustrates a branched IAC(1) in which IAC(1) includes an immune-inducing moiety (3) linked to affilin(21) via a first linker, and affilin(21) is optionally further linked to a chelating agent (91) via a further linker, and affilin(21) is specifically linked and / or attached to a further linker having HEAD(11). Thus, each linker may independently be the same as or different from any other linker in the molecule. However, in particular, the linker having HEAD(11) may be linked to affilin(21). Alternatively, IAC(1) may be a linear molecule, as represented by Examples 3H, 3J, and 3K, where the chelating agent (91) may be optionally present and linked to affilin(21) via a first linker, as shown in Figure 3H. Affilin (21) may be linked to IAM (3) via a second linker, and IAM (3) may be connected to and / or attached to a further linker having HEAD (11). In a further embodiment, as shown in Figure 3J, IAM (3) may be linked to affilin (21) which is connected to and / or attached to a further linker having HEAD (11) via a first linker. Furthermore, optionally, a linker (8) having HEAD (11) may link affilin (21) to a chelating agent (91).

[0241] In further embodiments, as shown in Figure 3KJ, IAM(3), which is connected to and / or attached to a further linker having HEAD(11), can be linked to affilin(21) via the first linker. Optionally, a linker(8) having HEAD(11) can link IAM(3) to a chelating agent(91).

[0242] In a further preferred embodiment, the subject is a mammal, preferably a human, that has cancer or is at risk of developing cancer.

[0243] In a particularly preferred embodiment, the immune inducer compound (IAC) according to the present invention comprises a target structure binding moiety and an immune inducer moiety (3), wherein the target structure binding moiety is linked to the immune inducer moiety (3) via a linker moiety (8), the target structure binding moiety can bind a target structure (4) of a target cell (5) and / or the target cell environment (6), the presence and / or overexpression of the target structure (4) indicates a target cancer disease, and the immune inducer moiety (3) can attract immune cells (7) to the target cell (5) and / or the target cell environment (6), the target structure binding moiety is a PSMA ligand and / or a PSMA-binding affilin (2, 21), and the immune inducer moiety (3) is the SARS-CoV-19 spike protein, particularly the RBD of the SARS-CoV-19 spike protein, or a part thereof. Thereafter, the IAC is preferably for the treatment of a cancer disease, the target structure (4) is PSMA, and the cancer disease is prostate cancer. In particular, the IAC, more specifically the linker (8), comprises a chelating agent (91), which is linked to a target structure binding moiety and / or an immune-inducing moiety (3). Thereafter, preferably, the chelating agent (91) complexes with a radioactive moiety (9), preferably gallium-68 (Ga-68) and / or lutetium-177 (Lu-177) or actinium-225 (Ac-225), or yttrium-90 (Y-90), or any other radiodiagnostic or radiotherapeutic isotope.

[0244] In a particularly preferred embodiment, the immune inducer compound (IAC) according to the present invention comprises a target structure binding moiety and an immune inducer moiety (3), wherein the target structure binding moiety is linked to the immune inducer moiety (3) via a linker moiety (8), the target structure binding moiety can bind a target structure (4) of a target cell (5) and / or the target cell environment (6), the presence and / or overexpression of the target structure (4) indicates a target cancer disease, and the immune inducer moiety (3) can attract immune cells (7) to the target cell (5) and / or the target cell environment (6), the target structure binding moiety is an FAP ligand and / or an FAP-binding affilin (2, 21), and the immune inducer moiety (3) is the SARS-CoV-19 spike protein, particularly the RBD of the SARS-CoV-19 spike protein, or a part thereof. Thereafter, the IAC is preferably for the treatment of a cancer disease, the target structure (4) is FAP, and the cancer disease is pancreatic adenocarcinoma. In particular, the IAC, more specifically the linker (8), comprises a chelating agent (91), which is linked to a target structure binding moiety and / or an immune-inducing moiety (3). Thereafter, preferably, the chelating agent (91) complexes with a radioactive moiety (9), preferably gallium-68 (Ga-68) and / or lutetium-177 (Lu-177) or actinium-225 (Ac-225), or yttrium-90 (Y-90), or any other radiodiagnostic or radiotherapeutic isotope.

[0245] In a particularly preferred embodiment, the immune attractant compound (IAC) according to the present invention comprises a target structure binding moiety and an immune attractant moiety (3), wherein the target structure binding moiety is linked to the immune attractant moiety (3) via a linker moiety (8), the target structure binding moiety can bind a target structure (4) of a target cell (5) and / or the target cell environment (6), the presence and / or overexpression of the target structure (4) indicates the cancerous disease of the subject, and the immune attractant moiety (3) can attract immune cells (7) to the target cell (5) and / or the target cell environment (6), the target structure binding moiety is a Her2 Neu ligand and / or a Her2 Neu-binding affilin (2, 21), and the immune attractant moiety (3) is the SARS-CoV-19 spike protein, particularly the RBD of the SARS-CoV-19 spike protein, or a part thereof. Thereafter, the IAC is preferably for the treatment of cancerous diseases. In particular, the IAC, more specifically the linker (8), comprises a chelating agent (91), which is linked to a target structure binding moiety and / or an immune-inducing moiety (3). Thereafter, preferably, the chelating agent (91) complexes with a radioactive moiety (9), preferably gallium-68 (Ga-68) and / or lutetium-177 (Lu-177) or actinium-225 (Ac-225), or yttrium-90 (Y-90), or any other radiodiagnostic or radiotherapeutic isotope.

[0246] In a second aspect, the present invention relates to a pharmaceutical composition comprising an immunoassay compound (IAC) according to a first aspect of the present invention.

[0247] In a preferred embodiment, the pharmaceutical composition is intended for use in the treatment, diagnosis, and / or prevention of cancer.

[0248] In a further preferred embodiment, the target cells are cancer cells, and the disease is a cancerous disease.

[0249] In a further preferred embodiment, the pharmaceutical composition includes a suitable carrier.

[0250] This is particularly suitable for embodiments in which the immune-inducing portion includes nucleic acids. In particular, since nucleic acids must remain stable in order to reach their target cells or the target cell environment, encapsulation of nucleic acids, such as mRNA molecules, is considered herein.

[0251] In a third aspect, the present invention relates to a method for treating cancer in a subject, Step a) of applying an immunoinducing compound (IAC) according to a first aspect of the present invention to the subject. This includes methods.

[0252] In a preferred embodiment, the subject is a mammal, preferably a human.

[0253] In a further preferred embodiment, the subject is suffering from or at risk of developing cancer.

[0254] In a further preferred embodiment, the method further includes the step of determining the initial immune state of a subject to at least one antigen, preferably the initial state of the immune response after vaccination of the subject.

[0255] In a further preferred embodiment, the method further includes the step of determining the “responsive” immune state of a subject to at least one antigen, preferably the response state of the immune response after treatment with IAC according to the present invention.

[0256] In a further preferred embodiment, the method further includes the step of vaccinating a target with an antigen contained in at least one immunoinducing portion of the immunoinducing compound (IAC) administered in step a).

[0257] In a further preferred embodiment, the method further includes a second step of determining the subject's immune status to at least one antigen, preferably determining the state of the immune response after vaccination of the subject.

[0258] In a further preferred embodiment, the method further includes the step of additionally vaccinating a target with an antigen contained in at least one immunoinducing portion of the immunoinducing compound (IAC) administered in step a).

[0259] In a further preferred embodiment, the method further includes a pre-targeting step, preferably performed one or two days before step a), which is performed before step a) in which an immunoinducing compound (IAC) according to any one of claims 1 to 90 is applied to the target.

[0260] In a further preferred embodiment, the method further includes the step of applying molecular imaging, preferably PET and / or PET / CT, before and / or after step a) of claim 95.

[0261] In a further preferred embodiment, the method further includes the step of applying it to a radionuclide.

[0262] In a further preferred embodiment, the method further includes the step of applying at least one immunoCP inhibitor selected from the group comprising inhibitors of CTLA-4, PD-1, PD-L1, 7-H3, LAG-3, TIM-3, VISTA, GITR, CD27, CD70, CD40, OX40, or 4-1BB.

[0263] In a further preferred embodiment, the method may use ipilimumab, tremelimumab, AGEN-1884, pembrolizumab, nivolumab, PDR001, SHR1210, semiprimab, REGN2810, pizilizumab, AMP 514, BGB A317, PF-06801591, AMP224, atezolizumab, durvalumab, avelumab, CK-301, BMS The process further includes applying an immunoCP inhibitor selected from the group including 936559, MGA-271, MGD-009, IMP-321, BMS-986016, LAG-525, TSR-022, MBG-453, CA-170, TRX-518, INCAGN01876, GWN-323, MEDI1873, MK-4166, MK-1248, BMS986156, Valrirumab, SGN-CD70A, ISF35, RO70097890, MEDI-6469, MOXR-0916, PF-04518600, MEDI-0562, Urelumab, and Utomirumab.

[0264] In particular, information on checkpoint inhibitors, as well as the types of drugs, the targets of each inhibitor, and the mAb isotypes, is shown in Table 1 below: [Table 1-1] [Table 1-2]

[0265] In a fourth aspect, the present invention relates to a method for inducing an immune response, comprising the step of applying an immune-inducing compound (IAC) according to a first aspect of the present invention to a subject, preferably a therapeutic method according to a third aspect of the present invention.

[0266] In a fifth aspect, the present invention relates to a method for determining the effectiveness of at least one treatment applied to a subject, wherein the treatment comprises step a) applying an immunoassay compound (IAC) according to any one of claims 1 to 90 to the subject, and the method for determining the effectiveness preferably comprises a molecular imaging step, which includes applying PET and / or PET / CT to the subject.

[0267] In a sixth aspect, the present invention relates to a method for determining the effectiveness of at least one treatment applied to a subject, preferably as described in claim 108, wherein the treatment comprises step a) applying an immune inducer compound (IAC) described in any one of claims 1 to 90 to the subject, and the method for determining the effectiveness comprises step of determining the induction of immune cells to and / or the target cell environment.

[0268] In a preferred embodiment, the number of immune cells in the subject is increased by at least 5% after step a) compared to before step a).

[0269] In a further preferred embodiment, the immunoinducing compound (IAC) according to the first aspect of the present invention is intended for use as a pharmaceutical.

[0270] In a further preferred embodiment, an immunoinducing compound (IAC) according to the first aspect of the present invention and / or a pharmaceutical composition according to the second aspect of the present invention are for use in the manufacture of pharmaceuticals for the treatment of cancer.

[0271] In a seventh aspect, the present invention relates to a method for producing an immunoassay compound (IAC) according to a first aspect of the present invention.

[0272] In a preferred embodiment, the method for manufacturing is: Step a) involves preparing at least one target structure-binding subprecursor molecule. Step b) to prepare at least one immune-inducing subprogenitor molecule, Step c) of linking the at least one target structure-binding partial precursor molecule to the at least one immune attraction partial precursor molecule comprises.

[0273] In a preferred embodiment, preferably according to the first aspect of the invention, the immune attraction compound (IAC) is produced by the method according to the seventh aspect of the invention.

[0274] In an eighth aspect, the invention relates to a nucleic acid molecule comprising a nucleic acid sequence encoding an immune attraction compound (IAC) or a part thereof, particularly a nucleic acid encoding an immune attraction part.

[0275] Materials and Methods Preparation of Immune Attraction Compound Synthesis of FAP (UAMC1110)-Covid19-spike full length and FAP (peptide precursor)-Covid19-spike full length In a first approach, in accordance with the invention, a Covid19-spike full length IAC specific for binding to the pan-tumor marker FAP as a target structure and as an immune attraction part (IAM) was synthesized.

[0276] Hereinafter, the inventors specifically describe two alternative approaches for synthesizing an IAC comprising an FAP ligand as a target structure-binding part and a Covid19-spike full length protein as an exemplary immune attraction part in accordance with the invention. It will be recognized by those skilled in the art that the IACs according to the invention obtained, FAP-Covid19-spike, and similarly other IACs according to the invention can be synthesized according to these and various other methods known to those skilled in the art.

[0277] Those skilled in the art will recognize UAMC1110 as one of the most commonly used FAP inhibitory proteins (see, for example, Front Chem 2021 Apr 14;9:640566.doi:10.3389 / fchem.2021.640566.eCollection 2021, Vab Rymenant et al.). Of course, other target structure binding moieties, particularly FAP-specific target structure binding moieties, can be used to implement the invention in its entirety and depending on the desired results, especially depending on the cancerous disease to be treated.

[0278] Those skilled in the art will also recognize, for example, sequences of FAP such as Sequence ID No. 9 or similar, disclosed in International Publication No. 2021005131 A1, as one of the most commonly used linear FAP peptide precursors that can be achieved by standard methods and general procedures for automated / semi-automated solid-phase synthesis known in the art (see, for example, European Patent No. 3763726 A1 for FAP peptides). Of course, other target structure binding moieties, particularly FAP-specific target structure binding moieties, can be used to implement the invention in its entirety and depending on the desired results, especially depending on the cancerous disease to be treated.

[0279] In the following, the inventors describe the functionalization of both UAMC1110 and the FAP peptide precursor with squalate (SA) or with NCS ester.

[0280] Functionalization of UAMC1110 with squalate (SA) Process 1: UAMC1110.SA The FAP precursors NH2-UAMC1110 (1.0 equivalent) and diethyl squareate (3.0 equivalents) were dissolved in phosphate buffer (0.5 M; pH 7; 0.5 mL) and stirred at room temperature for 2 hours. The pH of the reaction was adjusted, and if necessary, it was adjusted to pH 7-7.5 with sodium hydroxide solution (1 M). The product UAMC1110.SA (75%) was isolated by semi-preparation HPLC (column: Phenomenex Luna C18 (250 × 10 mm) 10 μm; flow rate: 5 mL / min; solvent: H2O / MeCN + 0.1% TFA) and obtained as a colorless solid after lyophilization.

[0281] Step 2: UAMC1110.SA.COVID19-Spike COVID19-spike (1.0 mg) was diluted with 0.5 M Na2HPO4 buffer (pH 9, 1 mL). A 10-fold molar excess of UAMC1110.SA solution (1 mg / mL) was added, and the pH was adjusted to 9 with 1 M NaOH solution. The mixture was shaken overnight at room temperature using a thermomixer. Subsequently, the resulting immunoattractant compound (IAC-UAMC1110.SA.COVID19-spike) was purified by fractionation SEC using a PD-10 desalting column (8.3 mL Sephadex® G-25) and PBS as the mobile phase.

[0282] Functionalization of UAMC1110 with NCS esters Process 1: UAMC1110.NCS NO2-UAMC1110 (1.0 equivalent) was dissolved in tetrahydrofuran (2 mL), and Raney® Nickel 2800® was suspended in the solution. Hydrogen was passed through the suspension, and it was kept under hydrogen at room temperature for 5 hours. After completion, the mixture was filtered through celite / sand, the celite was washed twice with methanol (5 mL), and the organic layer was concentrated under vacuum. The obtained product was used without further purification (approximately 90%). The residue was stirred in anhydrous dichloromethane (1 mL) and TEA at room temperature for 15 minutes. To this solution, thiophosgene (1.0 equivalent) dissolved in anhydrous dichloromethane (1 mL) was added, and the mixture was stirred for 1 hour. The solution was quenched with 1 M NaOH solution and extracted with dichloromethane. The organic layer was concentrated under vacuum, and the obtained product was dissolved in dichloromethane / trifluoroacetic acid (1:1, vol%). After 5 hours, the solvent was removed under vacuum, and the residue was purified by HPLC to obtain the product UAMC1110.NCS as a colorless solid (78%).

[0283] Step 2: UAMC1110.NCS.COVID19-Spike The COVID19 spike was incorporated into PBS at pH 7.4. Then, 60 μL of the solution was diluted to 1 mL with PBS at pH 7.4. The pH of the spike solution was raised to 8.8–9.0 with 0.1 M Na2CO3, after which 5 equivalents of UAMC1110.NCS in 15 μL of DMSO were slowly added. The reaction was incubated at 37°C for 1 hour, shaken at 300 rpm, and then purified by SEC and centrifugation to obtain the immunoattractant compound (IAC) IAC-UAMC1110.NCS.COVID19-spike.

[0284] SA-mediated functionalization of FAP peptide precursors Step 1: FAP peptide-NH2 synthesis The sequence Hex-Cys-Pro-Pro-Thr-Gln-Phe-Cys-OH was synthesized by Fmoc solid-phase chemistry on a trityl resin. After final resin cleavage and precipitation with cooled methyl-tert-butyl ether / cyclohexane (1 / 1), the crude peptide was lyophilized in water / acetonitrile. The crude peptide was dissolved in 10 mL of a 1:1 mixture of ethanol and acetonitrile. To this mixture, N,N-diisopropylethylamine and then 1,3,5-tris(bromomethyl)benzene (1.3 equivalents) were added. This solution was stirred for 1 hour, and then 2-aminoethanethiol (11.0 equivalents) was added. After 1 hour, the solvent was removed by evaporation, and the remainder was dissolved in acetonitrile and water (20 mL, 1:1 mixture, containing 50 μL of TFA). After lyophilization, the crude FAP peptide-NH2 was purified by reverse-phase high-performance liquid chromatography (RP-HPLC) (column: Phenomenex Luna C18 (250 × 10 mm) 10 μm; flow rate: 5 mL / min; solvent: H2O / MeCN + 0.1% TFA).

[0285] Step 2: Functionalization of FAP peptides with SA FAP peptide-NH2 (1.0 equivalent) and diethyl square acid (3.0 equivalents), which are FAP precursors, were dissolved in phosphate buffer (0.5 M; pH 7; 0.5 mL) and stirred at room temperature for 2 hours. The pH of the reaction was adjusted, and if necessary, it was adjusted to pH 7-7.5 with sodium hydroxide solution (1 M). The product FAP peptide.SA (73%) was isolated by semi-preparation HPLC (column: Phenomenex Luna C18 (250 × 10 mm) 10 μm; flow rate: 5 mL / min; solvent: H2O / MeCN + 0.1% TFA) and obtained as a colorless solid after lyophilization.

[0286] Step 3: FAP peptide.SA.COVID19-spike COVID19-spike (1.0 mg) was diluted with 0.5 M Na2HPO4 buffer (pH 9, 1 mL). A 10-fold molar excess of FAP peptide.SA solution (1 mg / mL) was added, and the pH was adjusted to 9 with 1 M NaOH solution. The mixture was shaken overnight at room temperature using a thermomixer. Subsequently, the resulting immune complex IAC-FAP peptide.SA.COVID19-spike was purified by fractionation SEC using a PD-10 desalting column (8.3 mL Sephadex® G-25, GE Healthcare) and PBS as the mobile phase.

[0287] Functionalization of FAP peptide precursors with NCSester Step 1: FAP peptide-NO2 The sequence Hex-Cys-Pro-Pro-Thr-Gln-Phe-Cys-OH was synthesized by Fmoc solid-phase chemistry on a trityl resin. After final resin cleavage and precipitation in cooled methyl-tert-butyl ether / cyclohexane (1 / 1), the crude peptide was lyophilized in water / acetonitrile. The crude peptide was dissolved in 10 mL of a 1:1 mixture of ethanol and acetonitrile. To this mixture, N,N-diisopropylethylamine and then 1,3,5-tris(bromomethyl)benzene (1.3 equivalents) were added. This solution was stirred for 1 hour, and then 2-nitroethanethiol (11.0 equivalents) was added. After 1 hour, the solvent was removed by evaporation, and the remainder was dissolved in acetonitrile and water (20 mL, 1:1 mixture, containing 50 μL of TFA). After lyophilization, the crude FAP peptide-NO2 was purified by reverse-phase high-performance liquid chromatography (RP-HPLC) (column: Phenomenex Luna C18 (250 × 10 mm) 10 μm; flow rate: 5 mL / min; solvent: H2O / MeCN + 0.1% TFA).

[0288] Step 2: Functionalization of FAP peptides by NCS The FAP peptide - NO2 (1.0 equivalent) was dissolved in tetrahydrofuran (2 mL), and Raney (registered trademark) Nickel 2800 (registered trademark) was suspended. Hydrogen was passed through the suspension and kept under hydrogen at room temperature for 5 hours. After completion, the mixture was filtered through celite / sand, the celite was washed twice with methanol (5 mL), and the organic layer was concentrated under vacuum. The obtained product was used without further purification (90%). The residue was stirred at room temperature for 15 minutes in dehydrated dichloromethane (1 mL) and TEA. To this solution, thiophosgene (1.0 equivalent) dissolved in dehydrated dichloromethane (1 mL) was added and stirred for 1 hour. The solution was quenched with 1M NaOH solution and extracted with dichloromethane. The organic layer was concentrated under vacuum, and the obtained product was dissolved in dichloromethane / trifluoroacetic acid (1:1, volume %). After 5 hours, the solvent was removed under vacuum, and the residue was purified by HPLC to obtain the product FAP peptide.NCS as a colorless solid (42%).

[0289] Step 3: FAP peptide.NCS.COVID19 - spike COVID19 - spike was incorporated into PBS at pH 7.4. Then, 60 μL of the solution was diluted to 1 mL with PBS at pH 7.4. After raising the pH of the spike solution to 8.8 - 9.0 with 0.1M Na2CO3, 5 equivalents of FAP peptide.NCS in 15 μL of DMSO were slowly added. The reaction was incubated at 37 °C for 1 hour with shaking at 300 rpm, followed by SEC and centrifugal filtration to purify the immune - attracting compound (IAC - FAP peptide.NCS.COVID19 - spike).

[0290] Synthesis of PSMA - Covid19 - spike full - length In a second approach, according to the present invention, a specific IAC with respect to binding to PSMA as a target structure and Covid19 - spike full - length as an immune - attracting moiety was synthesized. As described above, PSMA is of particular interest as a target structure specific to prostate cancer tumors.

[0291] Those skilled in the art will recognize amino-cyclohexa-naphthyl-Lys-CO-Glu-tri-O-tert-butyl ester (PSMA-NH2), which can be synthesized according to the literature (e.g., see https: / / doi.org / 10.1016 / j.ejmcr.2022.100084 European Journal of Medicinal Chemistry Reports Volume 6, December 2022, 100084, Kumar et al.), as one of the most commonly used PSMA inhibitory proteins. Of course, other target structure binding moieties, particularly PSMA-specific target structure binding moieties, can be used to implement the invention in its entirety and depending on the desired results, especially depending on the cancerous disease to be treated.

[0292] PSMA functionalization by SA Process 1: PSMA.SA PSMA-NH2 (1.0 equivalent) was dissolved in 0.5 M phosphate buffer (pH 7; 250 μL), and 3,4-diethoxycyclobuta-3-ene-1,2-dione (3.0 equivalents) was added. The pH was adjusted to pH 7 with 1 M NaOH solution, and the reaction solution was shaken overnight. After HPLC purification (column: Phenomex Luna C18 semi-preparation type (250 × 10 mm) 10 μL, flow rate: 5 mL / min, H2O / MeCN + 0.1% TFA), the product PSMA.SA (23%) was obtained as a colorless solid.

[0293] Step 2: PSMA.SA.COVID19-Spike COVID-19 spike (1.0 mg) was diluted with 0.5 M Na2HPO4 buffer (pH 9, 1 mL). A 10-fold molar excess of PSMA-NH2 solution (1 mg / mL) was added, and the pH was adjusted to 9 with 1 M NaOH solution. The mixture was shaken overnight at room temperature using a thermomixer. Subsequently, the resulting immune complex (IAC-PSMA.SA.COVID19-spike) was purified by fractionation SEC using a PD-10 desalting column (8.3 mL Sephadex® G-25, GE Healthcare) and PBS as the mobile phase.

[0294] PSMA functionalization by NCS Process 1: PSMA.NCS PSMA-NO2 (1.0 equivalent) was dissolved in tetrahydrofuran (2 mL), and Raney® Nickel 2800® was suspended in the solution. Hydrogen was passed through the suspension, and it was kept under hydrogen at room temperature for 5 hours. After completion, the mixture was filtered through celite / sand, the celite was washed twice with methanol (5 mL), and the organic layer was concentrated under vacuum. The obtained product was used without further purification (90%). The residue was stirred in anhydrous dichloromethane (1 mL) and TEA at room temperature for 15 minutes. To this solution, thiophosgene (1.0 equivalent) dissolved in anhydrous dichloromethane (1 mL) was added, and the mixture was stirred for 1 hour. The solution was quenched with 1 M NaOH solution, and extracted with dichloromethane. The organic layer was concentrated under vacuum, and the obtained product was dissolved in dichloromethane / trifluoroacetic acid (1:1, vol%). After 5 hours, the solvent was removed under vacuum, and the residue was purified by HPLC to obtain the product PSMA.NCS as a colorless solid (65%).

[0295] Process 2: PSMA.NCS.COVID19-Spike The COVID-19 spike was incorporated into PBS at pH 7.4. Then, 60 μL of the solution was diluted to 1 mL with PBS at pH 7.4. The pH of the spike solution was raised to 8.8–9.0 with 0.1 M Na2CO3, after which 5 equivalents of PSMA.NCS in 15 μL of DMSO were slowly added. The reaction was incubated at 37°C for 1 hour, shaken at 300 rpm, and then purified by SEC and centrifugation to obtain the immunoattractant compound (IAC-PSMA.NCS.COVID19-spike).

[0296] In the above example, the full length of the Covid19 spike was used. However, those skilled in the art will recognize that other proteins, particularly peptides, can similarly be used using techniques and methods known to those skilled in the art. For example, the RBD of the full length of the Covid19 spike can also be advantageously used as the IAM of the IAC of the present invention. The synthesis of other IACs, such as the HER2-Covid19-full length spike IAC, can be achieved in a similar manner. Thus, the use of affilin, in particular, may be advantageous and is considered to be within the scope of the present invention.

[0297] In particular with respect to Her2 neu, Her2-binding proteins can be obtained from the disclosure of International Publication No. 2017013129, which is inherited herein by reference.

[0298] Synthetic PSMA / FAP-DOTA-Covid19-Spike: Coupling with chelating agents Step 1: PSMA or FAP-peptide / UAMC1110-DOTA-Bn-pSCN To a mixture of pSCN-Bn-DOTA (1.0 equivalent) and HOBt (1.0 equivalent) in 30 mL of ethyl acetate cooled in an ice bath, DCC (1.1 equivalents) was added all at once. The mixture was stirred for 30 minutes. The formation of a white precipitate (DCU) was observed. Either PSMA-NH2, FAP peptide-NH2, or NH2-UAMC1110 (1.0 equivalent) and TEA (1.0 equivalent) were added. After stirring the mixture at room temperature for 2 hours, 15 mL of hexane was poured into the mixture. DCU was removed by filtration. The precipitate was washed with ethyl acetate. The combined filtrate was washed with 4% HCl (30 mL x 2), saturated NaHCO3 solution (25 mL x 3), and brine (30 mL x 3), and dried. HPLC purification yielded pSCN-Bn-DOTA-PSMA / FAP peptide / UAMC1110 as a colorless product (20-40%).

[0299] Step 2: PSMA / FAP peptide / UAMC1110-DOTA-Bn-SCN-COVID19-spike The COVID-19 spike was incorporated into PBS at pH 7.4. Then, 60 μL of the solution was diluted to 1 mL with PBS at pH 7.4. The pH of the spike solution was raised to 8.8–9.0 with 0.1 M Na2CO3, after which 5 equivalents of PSMA / FAP peptide / UAMC1110-DOTA-Bn-pSCN in 15 μL of DMSO were slowly added. The reaction was incubated at 37°C for 1 hour, shaken at 300 rpm, and then purified by SEC and centrifugation to obtain the immunoattractant compound (IAC PSMA / FAP peptide / UAMC1110-DOTA-Bn-SCN-COVID19-spike).

[0300] Labeling with radioactive metals The molecules obtained above are further labeled with a radioactive metal.

[0301] Ga-68 was eluted from a 68Ge / 68Ga generator (GalliaPharm®, Eckert & Ziegler Radiopharma GmbH, Germany) and used without further purification. Radiolabeling was performed at 85°C using 2.0 mL of 0.7 M ammonium acetate buffer pH 5.5 and 50 μg of precursor with an All-In-One mini synthesis module (Trasis, Belgium) equipped with disposable cassettes. Purification was performed using a size exclusion column (SEC). The product was then diluted with 10 mL of physiological saline and passed through a sterile filter (Sterifix®, B. Braun SE, Germany).

[0302] [ 177Lu]LuCl3 non-carrier added (nca) (EndolucinBeta®) was obtained from ITM Isotope Technologies Munich (Germany) at different radioactivity levels, dissolved in 0.04 M HCl (40 GBq / mL), and used without further purification. Radiolabeling was performed at 85°C using a synthesis module equipped with disposable cassettes, with 2.0 mL of 0.15 M ascorbic acid buffer pH 4.5 and 37-50 μg of precursor / GBq Lu-177. Purification was performed using a size exclusion column (SEC), diluted in 15 mL of physiological saline, and formulated by passing through a sterile filter (Sterifix®, B. Braun SE, Germany).

[0303] In all of the above approaches, pH was controlled at the start and after labeling using disposable pH stripes (Merck, Germany). For reaction control, a radio-TLC (TLC Silica gel 60 F254 Merck, Germany) was used with (1) citrate buffer pH 4 and (2) a 1:1 mixture (v:v) of 1.0 M ammonium acetate buffer and MeOH as the mobile phase.

[0304] Analytical radioHPLC was performed using an Agilent Infinity 1200 HPLC system (100 μl injection loop, 20 μl injection for quality control) equipped with a Ramona* radioactivity detector (Elysia-Raytest, Belgium) and an external BGO scintillator flow cell (300 μL, Elysia-Raytest, Belgium). The following column, BioSepSEC LC Column 300 × 7.8 mm, Phenomenex, Germany, was used. The mobile phases were acetonitrile (A) and water (B), each containing 0.1% TFA. A gradient at a flow rate of 1.0 mL / min was performed, starting with 5% A and increasing to 45% A within 10 minutes. For data interpretation, TLC was analyzed using a miniGita TLC scanner (Elysia-Raytest, Belgium) and analytical software Gina (Elysia-Raytest, Belgium).

[0305] Stability studies were performed in a triplicate at 37°C in human serum (HS), physiological saline, and the final product formulation. HS (human male AB plasma, US origin) was obtained from Sigma Aldrich, USA. Physiological saline was purchased from B. Braun, Germany. The final step involved using 100 μL of the formulated product solution (20 MBq) added to either 1 mL of HS or physiological saline. Radio-TLC was performed under the analytical conditions described above.

[0306] Determination of binding specificity In this field, various target structure binding moieties / target structure pairs are known. The target structure binding moieties can bind to the target structure of the target cell, and the presence and / or overexpression of the target structure indicates the target cancer disease, as summarized in Table 2 below. [Table 2]

[0307] From the above, it is scientifically proven that each target structure (the presence and / or overexpression of the aforementioned target structures indicates the target cancer disease) is specifically bound by its respective target structure binding site.

[0308] To demonstrate that the target structure binding portion of the synthesized immunoattractant compound (IAC) according to the present invention can bind to the target structure of target cells (4), the binding specificity can be determined, and various methods are known in the art, in particular, including immunoprecipitation (IP), fluorescence resonance energy transfer (FRET), surface plasmon resonance (SPR), ELISA (enzyme-linked immunosorbent assay), in vivo imaging, and Biacore Technology, especially in appropriate animal models.

[0309] Radiocompetitive ligand binding assay In this experiment, we will use Chinese hamster ovary cells (CHO), CHO-PSMA, CHO-FAP, or CHO-HER2 cell lines that overexpress each receptor / surface protein to determine the binding affinity of 68-Ga-PSMAi-IAM, 68-Ga-FAPI-IAM, or 68-Ga-Her2 / neu-IAM to human PSMA / FAP / Her2 in a competitive binding study.

[0310] For example, to assess binding affinity, a competitive binding assay was performed as follows: CHO-PSMA cells overexpressing prostate-specific membrane antigen (PSMA) were cultured in Dulbecco's modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum and 1% penicillin / streptomycin. The cells were maintained at 37°C in a humidified atmosphere of 5% CO2. 68-Ga-PSMAi-IAM (25 μL) and increasing concentrations (10 -10 )~10 -6 A solution containing a mixture of 68-Ga-PSMAi-IAM and an unlabeled DOTA conjugate anti-PSMAi (25 μL, as a competitor) was added to CHO-PSMA cells (400,000 cells per vial). After incubation at 37°C for 1 hour, the cells were centrifuged at 600 g (1,200 rpm; Biofuge 15) for 5 minutes. The supernatant was carefully removed from each vial, followed by two washes of cells with 250 μL of PBS. The washes were combined with the previously removed supernatant, and the supernatant represented the amount of free radioligand. Subsequently, the amount of radioactivity bound to cells (cell pellet) and the amount of free radioligand were quantified using a 2470 Wizard2 γ-counter (PerkinElmer). Given the high structural similarity between 68-Ga-PSMAi-IAM and the unlabeled ligand, it is considered that they exhibited nearly identical affinity for PSMA, resulting in homologous competitive binding. To verify specific binding, transduced CHO cells were used as a negative control.

[0311] In vivo coupled assay / comparison 68-Ga-PET / CT To prepare the xenograft model, 5 athymoid nude mice aged 6-8 weeks were subcutaneously injected into the right flank with 5 × 10^6 CHO-PSMA or CHO-control cells suspended in 100 μL of Matrigel. 68-Ga-PSMAi and 68-GA-PSMAi-IAM were synthesized using standard procedures or as described above. The xenograft was 150-200 mm. 3 Once the mice reach their average size, they are subjected to PET / CT imaging. Prior to imaging, the mice are anesthetized with isoflurane and a tail vein catheter is inserted for intravenous radioactive tracer injection. A preclinical small animal PET / CT scanner is used for PET / CT imaging. The mice are placed in a supine position within the scanner, and a CT scan is performed first for anatomical reference. Subsequently, 5–10 MBq of 68-Ga-PSMAi in a volume of 100 μL is injected intravenously, and dynamic PET imaging is performed for a predetermined period until complete elimination. Two days later, a PET / CT scan is performed again using 10 MBq of 68-Ga-PSMAi-IAM. The mice are euthanized under anesthesia, and organs are collected for in vivo distribution analysis. The PET images are reconstructed and simultaneously registered with the CT images for anatomical localization. Regions of interest (ROIs) corresponding to areas suspected of specific binding are manually selected on the acquired images. These ROIs are defined based on anatomical (xenograft) and / or functional criteria (target gene expression). ROIs were visualized by experienced radiologists using MIM software (MIM software Inc., 2014) to ensure consistency and accuracy in quantifying radiotracer uptake in xenografts. Standardized uptake values ​​(SUVmax) were calculated to determine the specific binding of the 68-Ga-PSMA vs. 68-Ga-PSMAi-IAM tracer to PSMA-expressing xenografts. Non-transferred CHO cells were used as a negative control group to assess non-specific tracer uptake.

[0312] SUV cost quantification Within the selected ROI, the standardized inclusion value (SUV) was calculated. The maximum SUV (SUVmax) and average SUV (SUVmean) were determined using the following formulas: SUVmax = (Radioactivity concentration in ROI [Bq / mL]) / (Injected dose [Bq] / Body weight [g]) SUVmean = (Total radioactivity in ROI [Bq]) / (ROI volume [mL] * injected dose [Bq] / body weight [g])

[0313] Clinical process 1: Patient selection (SelofP) Patients who may be candidates for the treatments and methods of the present invention, and who can therefore be successfully treated with the immunoinducing compounds of the present invention, are diagnosed with malignant tumors by standard methods known in the art (Table 2). For example, patients who are positive on a corresponding PET scan and / or positive on a biopsy by quantitative qPCR or immunohistochemistry may be selected for treatment and subsequently subjected to the methods and treatments described below. In particular, any specific diagnostic method accepted in the art, for example, whose results indicate the presence of the cancerous disease in question, is appropriate.

[0314] SelofP Criterion 1: Tumor entity and target gene expression For example, according to the current ESMO guidelines for prostate cancer diagnosis (https: / / doi.org / 10.1016 / j.annonc.2020.06.011), patients with prostate cancer can be identified, for example, by elevated PSA levels (PSA >1 ng / ml at age 40 or >2 ng / ml at age 60), positive multiparametric magnetic resonance imaging (mpMRI), and / or positive histopathological or immunohistopathological evaluation of tumor biopsy (Gleason score >6). Patients suitable for the invention presented herein may be identified by positive Ga-68-PSMA-PET / CT scan or positive Ga-68-FAPI-PET / CT, and the ideal SUVmax cutoff value will be assessed during clinical trials as shown in Table 3. [Table 3]

[0315] As an alternative or additional method to verify the expression of a target gene, qPCR or immunohistochemistry can be performed, and this can also be advantageously performed in a revised order, i.e., markers and diagnostics first, followed by PET.

[0316] Analysis of target gene expression using qPCR Nucleic acids are extracted from 1 ml of whole blood EDTA and heparinized samples. RNA is extracted from whole blood samples according to the general instructions for using a commercially available bead-based extraction method (Xtrakt kit; Stratifyer Molecular Pathology GmbH, Cologne, Germany). Briefly, 100 μl of blood or plasma is placed in a 1.5 ml microcentrifuge tube, 100 μl of erythrocyte lysis buffer is added, and the mixture is incubated in a thermomixer at 95°C for 15 minutes with shaking at 100 rpm. Then, the solubilizer is treated with proteinase K at 65°C for 15 minutes. Subsequently, binding buffer and magnetic beads are added, and the mixture is incubated at room temperature for 15 minutes with shaking at 1200 rpm. The supernatant is discarded, and the beads are washed by a three-cycle process of adding wash buffer, aspirating, magnetizing, and discarding the supernatant. Finally, add 100 μl of elution buffer and dilute the nucleic acid by incubation at 95°C for 15 minutes with shaking at 1000 rpm. Then, magnetize the beads and digest the supernatant with DNAse I to obtain total RNA from the sample without DNAse. The RNA eluate is then stored at -80°C until use.

[0317] mRNA levels of PSMA, FAP, and Her-2, as well as the reference genes calmodulin 2 (CALM2) and beta-2 microglobulin (B2M), were determined by one-step RT-qPCR using the SuperScript III RT-qPCR system (Invitrogen, Waltham, Massachusetts, USA) and gene-specific primer-probe combinations (assay numbers, respectively; STRATIFYER Molecular Pathology GmbH, Cologne, Germany).

[0318] In accordance with the MammaTyper® Instruction Manual 140603-90020-EU Rev 2.0, each patient sample or control was analyzed in pairs using a Light Cycler LC480 Instrument II (Roche Diagnostics, Rothkreuz, Switzerland) in 40 cycles of 30 minutes at 50°C, 2 minutes at 95°C, followed by 15 seconds at 95°C and 30 seconds at 60°C, as per the manufacturer's instructions. Gene expression was quantified using a modified method by Schmittgen and Livak by calculating 40-ΔCt, where ΔCt is calculated as the difference in Ct between the mean of the test gene and the reference gene (23). Gene expression levels were calculated as described above. : In short-cycle quantification, the threshold (Cq) value of the marker gene (MG) for each sample (S) was estimated as the median of three measurements. To compensate for inter-run variability, the Cq value is normalized to the mean expression of the REF gene and offset against the calibrator (PC) (ΔΔCq method). By subtracting ΔΔCq from the total number of cycles

[40] , it is ensured that the normalized gene expression is proportional to the corresponding mRNA expression level. This method facilitates the interpretation of data and clinicopathological correlations. The various calculation steps are summarized in the following equation: 40 - ΔΔCq(MG)S = 40 - ((Cq[MG]S - mean Cq[REF]S) - (Cq[MG]pc - mean Cq[REF]pc)). Finally, the individual biomarker dynamics during ICI treatment are determined by subtracting the post-treatment expression value from the pre-treatment expression value (Δ expression).

[0319] Analysis of target gene expression by immunohistochemistry: In short, immunohistochemical staining for the following proteins—anti-PSMA, anti-FAP, and anti-Her2—is performed on 4 μm formalin-fixed paraffin-embedded (FFPE) tissue sections from core needle biopsies using a Ventana Benchmark Ultra automated staining system (Ventana) at a diagnostic immunohistochemistry core facility. Positive signals are evaluated by a specially trained pathologist.

[0320] SelofP Criterion 2: Pre-treatment immune response Cancer-positive patients suitable for treatment either have a pre-existing immune response to IAC / IAM before treatment, or are vaccinated or boosted with IAC / IAM before initiating treatment to build an active immune response to IAC / IAM.

[0321] For example, antibody titers against the Covid-19 spike, which is the immune-inducing portion, are assessed in baseline serum samples from treatment-naive patients. Therefore, SARS-CoV-2 specific antibodies are assessed using antibody-specific enzyme-linked immunosorbent assay (ELISA) or alternative serological techniques, for example, using the Elecsys® anti-SARS-CoV-2 S assay on cobas e411 (Roche Diagnostics). A titer of 0.8 U / ml or higher in the Elecsys® Anti-SARS-CoV-2 S-Assay is considered positive and indicates a pre-existing immune response.

[0322] Clinical step 2: Pre- and post-treatment analysis of the patient's immune response status and TME composition. 1. To assess the pre-treatment anti-tumor immune response in the blood, and 2. To assess the immune cellular status in the tumor's tissue microenvironment, an immune baseline test is performed. An increase in tumor immune-infiltrating cells and increased cytokine secretion from pre-treatment to post-treatment is considered to indicate successful immunotherapy. Therefore, the respective tests are performed before and after the initiation of treatment (at the time points shown below): a) Immunotherapy phenotyping of peripheral blood mononuclear cells (PBMCs) is performed using fluorescence-guided cell sorting (FACS) in whole blood samples treated with anticoagulation using EDTA or heparin, with a panel of immunocytoplasmic markers (e.g., CD3, CD8, CD56, CD16, CXCR3, CCR4, CCR2, CCR5, CXCR6, PD-1, PD-L1, CTLA-42, CD45RA, CD45RO, CD62L, CCR7). b) CRP and cytokine levels (e.g., IL2, IL12, IL15, IL21, INFγ, TNFα) in serum or PBMCs are analyzed by ELISA or quantitative PCR (qPCR). c) FFPE tissue material from fresh biopsy primary tumor specimens of treatment-naïve biopsies is analyzed by multiplex immunohistochemistry for tumor-infiltrating immune cells and molecular subtypes, hormone receptor status, repair gene mutation status (BRCA1 / 2, CHEK2, etc.), and apoptosis / necrosis / ferroptosis markers. (Multiplex immunohistochemistry accurately defines the immune status of metastatic melanoma | Scientific Reports (nature.com)) d) mRNA or FFPE analysis from fresh tissue material using the Nanostring® nCounter platform with the PanCancer IO 360® panel.

[0323] CRP and cytokine levels Baseline CRP and cytokine levels are determined before and after treatment using a standardized laboratory system derived from clinical routines, such as ELSIA or qPCR.

[0324] mRNA extraction is performed as described above. mRNA levels of IL2, IL12, IL15, IL21, INFγ, TNFα, CD3, CD8, PD1, PD-L1, CTL1-4, and the reference genes calmodulin 2 (CALM2) and beta-2 microglobulin (B2M) are determined by one-step RT-qPCR using the SuperScript III RT-qPCR system (Invitrogen, Waltham, Massachusetts, USA) and gene-specific primer-probe combinations (assay numbers, respectively; STRATIFYER Molecular Pathology GmbH, Cologne, Germany). Analysis of the qPCR results is performed as described above.

[0325] Assessment of the identity and quantity of immune cells by single or multiple immunohistochemistry. To determine the immunological status of tissue samples, nucleic acids are extracted from 4 μm FFPE tissue sections obtained from pre-treatment tumor samples and post-treatment biopsies using a commercially available kit (RNXtract; STRATIFYER Molecular Pathology GmbH, Cologne). In short, at the inventors' diagnostic immunohistochemistry core facility, 4 μm FFPE tissue sections on a Ventana Benchmark Ultra automated staining system (Ventana) are immunohistochemically stained for the following proteins: CD3 (F7.2.38, monoclonal mouse, ThermoFisher-Scientific®, dilution 1:50), CD8 (C8 / 144B, mouse monoclonal, ThermoFisher-Scientific®, dilution 1:50), CD68 (PG-M1, mouse monoclonal, ThermoFisher-Scientific®, dilution 1:60), PD-1 (NAT105, mouse monoclonal; Ventana), and CD56 (MRQ-42, monoclonal mouse, CELL MARQUE®, dilution 1:50), and PD-L1 (SP263 assay, Ventana).

[0326] Spatial multiplexed immunohistochemistry Multiple immunohistochemistry is performed on the Phenocycler Fusion Platform using the PhenoCode™ Discover Immune Profiling Human Protein Core, which contains antibodies against CD3e, CD4, CD8, CD11c, CD14, CD20, CD44, CD45, CD45RO, CD56, CD68, HLA-A, HLA-DR, Ki67, and pancytokeratin.

[0327] The results of single and multiple immunohistochemistry are evaluated by two expert pathologists guided by automated image analysis (Akoya Systems).

[0328] Automated image analysis using Definiens Developer Software As previously described, assess tumor tissue, normal tissue and stromal tissue, as well as empty spaces

[19] . Quantify CD3+, CD8+, CD68+, PD-1+, and CD56+ ICs (counts / mm2) and convert to log2 for further analysis.

[0329] mRNA expression analysis of TME composition mRNA is extracted from FFPE or fresh-frozen tumor biopsies as described above. Using a hybridization-based digital counting assay, specifically the Nanostring® nCounter platform, purified total RNA from treatment-naive, treated, and control cohorts is subjected to a PanCancer IO 360® panel to measure 770 genes related to immunity and control genes. Normalized and log2-converted expression is assessed to identify immune cell abundance, immune signaling, tumor characteristics, and stromal biological signatures.

[0330] Clinical step 3: Diagnosis and / or treatment of the patient in any of the following ways A) 68-Ga labeled PSMAi / FAPi / Her-2i-IAM or for diagnostic and immunotherapy B) For combination immunotherapy / theranostic therapy 177Lu-PSMAi / FAPi / Her-2i-AIC C) Label-free PSMAi / FAPi / Her2i-IAM for immunotherapy

[0331] The application is intended to induce an increase in relevant immune cells during TME and to effectively prompt the immune system to attack tumor cells linked to selected immune-inducing regions.

[0332] A) 68-Ga labeled PSMAi / FAPi / Her-2i-IAM or for diagnostic and immunotherapy Safety, internal distribution, and dose measurement assessment of 68-Ga-PSMAi / FAPi / Her2-IAM In Phase I clinical studies, safety, intracellular distribution, and dose measurement are the primary objectives of assessment. A secondary objective is to investigate the potential for tumor targeting.

[0333] Based on the criteria outlined in SelofP, 20 patients, exemplified in Table 4 below, received injections of 68-Ga labeled PSMAi / FAPi / Her2-IAM tracers and 150-200 MBq of 68-Ga-PSMAi / FAPi / Her2-IAM v. injections. The radioactivity of the administered radioisotopes was within the range of 150-200 megabecquerels (MBq). Dosimetry assessments were performed by PET / CT scans at three time points after injection, specifically at 10, 60, and 90 minutes. Physical examinations and blood analyses were performed to ensure safety. Dosimetry calculations were performed using MIM (MIM software Inc., 2014) for intracellular distribution analysis for 11 different organs and regions of interest (ROIs), while dosimetry calculations were performed using OLINDA / EXM software 1.0 (Organ Level Internal Dose Assessment / EXponential Modeling, Vanderbilt University). The singularity of a bond is calculated by the ratio of SUVmax and SUVmean in the ROI for the region with the smallest or lowest tracer bond reference region.

[0334] The patient cohort is classified into three subgroups, each corresponding to a different administered mass of the 68-Ga-PSMAi / FAPi / Her2-IAM tracer: 0.01 milligrams for patients 1-7, 0.1 milligrams for patients 8-15, and 0.5 milligrams for patients 16-20. This stratification enables investigation of potential variations in normal biodistribution. The goal is to assess whether increasing the tracer mass leads to a decrease in non-specific binding in non-target organs. Importantly, the radioactivity of the administered tracer is consistent across all patient groups, ranging from 50 to 200 MBq.

[0335] Safety assessment Measure vital signs including blood pressure, heart rate, and body temperature and perform additional clinical laboratory assessments including standard hematological and comprehensive metabolic panels. These panels included measurements of hemoglobin, white blood cell count, neutrophils, lymphocytes, platelets, creatinine, blood urea nitrogen, calcium, sodium, potassium, carbon dioxide, lactate dehydrogenase, alanine transaminase, aspartate aminotransferase, alkaline phosphatase, total bilirubin, and albumin.

[0336] These evaluations are performed both pre-injection and 2 hours post-injection of the compound. Subjective adverse experiences are also systematically monitored, using open-ended questions to assess the patient's experience at pre-injection, during the 2-hour period spent in the nuclear medicine department, and by telephone follow-up observations up to 24 hours post-injection.

[0337] B) For combination immunotherapy / seranostic 177 Lu-PSMAi / FAPi / Her-2i-AIC The application is for inducing an increase in relevant immune cells in the TME and for effectively attacking tumor cells linked to the selected immune-attracting moiety by the radioisotope 177The addition of Lu is thought to enhance the theranostic effect due to direct radiation damage to tumor cells and the abscopal effect on the immune response. The abscopal effect has been described in immunotherapeutic approaches as the ability to induce localized, distant activation of tumor cells via radiation.

[0338] example: At a dose of 5 GBq 177 Intravenous injections of Lu-PSMAi-IAM were repeated every six weeks.

[0339] Typically 1-5 GBq, administered to the target group at an appropriate location or intravenously. 177 Lu is administered. Here, different groups can be treated with doses that have been proven effective for each IAC molecule, depending on its size and molecular weight.

[0340] To assess immune cell infiltration mediated by the application of IAC, the analysis outlined in Clinical Step 2 is performed 2–5 days, and / or blood is taken daily for 7 days, then weekly, and a biopsy is performed 2 months later and compared to treatment-untreated samples.

[0341] If necessary, clinical step 3 may be repeated with radioisotope-containing / non-containing IACs after imaging indicates the disappearance of the immune-inducing compound, until the associated immune response can be observed. Additionally or alternatively, measurements of immune parameters may be repeated to confirm whether they have increased. Re-staging may also be considered to assess the progress of treatment.

[0342] C) Application of immunotherapy agents without radionuclides. 68 Molecular imaging via Ga or 177 Instead of step 3 above and the first alternative step 3, which are primarily used for diagnostic and therapeutic purposes via radionuclide therapy using Lu, the IAC according to the present invention can also be applied without loading a radionuclide.

[0343] Therefore, the patient group is treated with 20-150 mg of IAC, either locally or intravenously, taking into account the molecular weight differences of the different IAC molecules. Treatment can be repeated with similar doses approximately 2-4 weeks later.

[0344] Similar to step 3) or the first alternative step 3), an imaging method can be used to determine the effectiveness of binding of the target structure binding moiety to the target structure and can be compared with the baseline data determined in step 1. In this way, the effect of the immunoattractant compound on the binding constant and mean binding time can be assessed.

[0345] If necessary, and after imaging indicates the disappearance of the immune-inducing compound, alternative step 3 may be repeated until the relevant immune response can be observed or until the recommended maximum radiation dose is achieved.

[0346] An effective immune response can be determined by an increase in serum CRP levels (doubling of individual baseline levels during the initial treatment period / "flare responder" assessment), an increase in individual CD8 mRNA levels (at least a difference of 1 DCT value higher is equivalent to a doubling of relative cell type-specific transcript levels), or an increase in body temperature (above 38°C). The antitumor efficacy obtained is Ga 90 PSMA imaging or Ga 90 It can be quantified by repeated conventional radioligand assessments using PSMA-IAC imaging.

[0347] Those skilled in the art will recognize that the application of lutetium (or other β-emitter) chelates, by application of immunoassay compounds, actually increases the levels of T cells, B cells, and macrophages. Therefore, new molecular entities produced by radiation face significantly increased response levels during time, and thus amplify the immune response.

[0348] Step 4: Immune response assessment a) As demonstrated in our previous research (Brubeck et al., Transl Lung Cancer Res 2021 | https: / / dx.doi.org / 10.21037 / tlcr-21-587), dynamic changes in immune cell markers such as CD3 and CD8 levels during cancer treatment indicate a significant extension of average lifespan when immunomodulatory agents are applied, as exemplified by the first-line monotherapy application of the checkpoint inhibitor pembrolizumab (Keytruda®) in metastatic NSCLC.

[0349] b) Monitor CD3 and CD8 levels in the blood at regular intervals over the course of treatment and up to 6 months post-treatment by molecular means (e.g., RT-qPCR of nucleic acid extracts from whole blood samples) or fluorescence-activated cell sorting (FACS) analysis using cell type-specific antibodies along with fluorescent measurement markers. An increase in CD3 / CD8 levels in individual patients is expected to indicate induction of an immune response and subsequent systemic tumor destruction, including micrometastatic lesions not visible at the time of immunotherapy. Increases in individual immune cell levels are determined, preferably by comparing at least two different blood samples at pre-treatment and post-treatment, or at two different time points post-treatment. When RT-qPCR is applied to nucleic acid extracts from blood samples, changes in candidate gene expression (e.g., CD3, CD8, CD69, PD-1) normalized to a reference / housekeeper gene (e.g., CALM2, B2M, etc.) at two time points are determined by the 40-DCT method, where higher numbers reflect higher gene expression levels. An increase in expression is determined by subtracting the relative gene expression from the second blood sample (preferably at the baseline level) from the first blood sample (preferably at the baseline level). An increase can be described if the difference is greater than 0. Most preferably, to account for technical variability, the increase is greater than 0,3DCT value.

[0350] c) As the inventors have published on immunomodulatory therapy in advanced bladder, kidney, and lung cancer (see, for example, Kluemper et al., 2022 May.;167:13-22.doi:10.1016 / j.ejca.2022.02.022), determining CRP levels using standard laboratory procedures is also effective in determining a "flare" of temporarily increased inflammatory levels within the first two weeks after treatment, which predicts the response to checkpoint inhibitors and improved progression-free survival (PFS) and overall survival (OS). For CRP flare responders, the predicted CRP dynamics at treatment are defined as at least a doubling of baseline CRP within the first month after initiation of checkpoint therapy, followed by a decline below baseline within the next three months. The same definition is applied to assess the effectiveness of immunotherapy.

[0351] d) Determination of serum antibody titers (against the immunization inducer) over the course of treatment. Determination of the antibody threshold level. Antibody levels are expected to increase compared to booster immunization vaccination. An increase in antibody titer of at least 50% can be used to determine the responder to immunization inducement therapy.

[0352] e) If necessary, biopsies can be performed at appropriate locations to observe any relevant changes in the TME (macrophage, T, or B cell count). This can be achieved by subjective assessment of HE scans or histopathological assessment of intratumoral lymphocytes by AI-based analysis. Alternatively, comparative analysis of IHC staining of immune cell markers (CD3, CD8, CD68, CD69, etc.) can be used to determine the increase in infiltrating immune cells in the target lesion. Similarly, mRNA can be extracted from FFPE tissue sections using commercially available kits to determine the state of immune cell infiltration and cellular composition of the tumor stroma, and assessed by standard RT-qPCR assays (STRATIFYER Milecular Pathology GmbH, Cologne) for candidate genes such as CD3, CD8, CXCL9, and IFN gamma. As the inventors have previously published, these methods are useful for distinguishing "cold" tumors from "hot" tumors and are beneficial for assessing individual prognoses (see, for example, Pfannstiel et al. Cancer Immunol Res 2019 Jun;7(6):923-938.doi:10.1158 / 2326-6066.CIR-18-0758.; Erlmeier et al. Eur Urol 2023 Feb;83(2):133-142.doi:10.1016 / j.eururo.2022.10.020.).

[0353] Tumor size is expected to shrink or disappear over the course of treatment, as it can be quantified by comparing SUV at two (to) time points using molecular tumor volume assessment, radioligand-mediated in vivo imaging, and the respective immune inducers themselves, in correlation with the immune response.

[0354] From the literature (Brueckl et al., Transl Lung Cancer Res 2021 | https: / / dx.doi.org / 10.21037 / tlcr-21-587), it is known that dynamic changes at the CD3+ and CD8+ cell levels during cancer treatment significantly extend average lifespan.

[0355] Blood CD3+ and CD8+ cell levels are monitored at regular intervals throughout the treatment period and up to 6 months post-treatment. CD3+ / CD8+ cell levels increase in individual patients who are systemically responding to combination therapy with radioactive ligands and immune inducers. The number of tumor-infiltrating immune cells increases due to the increased recognition of immune inducers. Antibody binding from previous vaccination or infection with the respective immunogenic factors of coronavirus leads to the direct accumulation and activation of immune cells with Fc receptors, as well as the subsequent secretion of immune activators, as can be determined by the increase in IFNG and CRP levels in the peripheral blood. Simultaneously, the internalization of radioactive immune inducers leads to cell destruction and exposure of intracellular components to phagocytic immune cells, accompanied by the subsequent presentation of digested protein fragments on MHC I and / or MHC II molecules. This leads to the activation of the respective T cells and their subsequent activation via T cell receptor recognition. The maturation of naive T cells into CD8-positive cells and their proliferation result in elevated T cell levels above baseline levels, which can be detected by FACS analysis or RT-qPCR of each immune cell-specific transcript from nucleic acid extracts of whole blood samples before and after treatment. Typically, a doubling of serum CRP levels, an increase in CD8 mRNA levels, or an increase in CD8 cell count indicates that active determination of antibody titers (against the inducer) in serum over the course of treatment can also be used to detect effective immune inducer therapy. Determination of antibody threshold levels can be performed by standard methods used to assess effective vaccination against COVID-19. An increase in antibody levels compared to booster vaccination is expected to indicate successful antitumor therapy with the respective immune inducer, with an increase of at least 20% indicating a response to treatment in the target lesion.

[0356] If necessary, biopsies can be performed at appropriate sites to observe any relevant changes (macrophage, T, or B cell counts) during TIME. Here, increased mRNA expression of the chemokine CXCL9 indicates T cell recruitment to the target lesion. This is also evident by an increase in immune cell infiltration into the tumor stroma, thereby transforming the “cold” tumor into an inflammatory tumor. This is accompanied by a dynamic increase in PD-L1 expression by tumor cells and neutrophils. This initial immune cell enrichment at the target site occurs as early as a few hours after radioligand / immune inducer combination therapy. The degree of the immune response can be determined by an increase in body temperature (>37°C, typically 38-39°C) and particularly nocturnal sweating, as well as by time-dependent digital devices such as wearables or by conventional temperature assessments by healthcare professionals at the treatment facility.

[0357] Efficacy research Various criteria can be used to assess the effectiveness of treatment with IAC according to the present invention, and in particular, potential criteria for treatment response include the following: - Progression-free survival -Overall survival -> Good physical condition score> - Relief of tumor-related symptoms These can be assessed using, for example, the following: RECIST v1.1 blinded independent central review (BICR); imaging using standard tracer-PET-CT (total tumor glucose metabolism (TLG), metabolic tumor volume (MTV), and standardized uptake (SUV), MRI (volume), ultrasound (volume), spectral-CT (TLG, MTV, SUV)). Metabolic changes in tumors as detected by 18-F-FDG-PET DW-MRI, DCE-MRI (Apparent diffusion coefficient (ADC) value)

[0358] Regarding immunotherapy, RECIST v.1.1 may not be applicable due to pseudo-exacerbations, and therefore irRECIST - Decrease in biomarker levels - Prostate cancer: Decrease in PSA (PSA response rate, PSA PCWG3 guideline waterfall plot)

[0359] Furthermore, it is possible to monitor molecular tumor volume. Various methods known in the art can be applied to determine the effectiveness of treatment for subjects requiring treatment using IAC according to the present invention. In particular, determining the molecular tumor volume over time is considered for monitoring the treatment effect. The molecular volume of the tumor can be determined in particular using imaging techniques such as PET / CT scans.

[0360] This allows for the clear visualization of tumor boundaries through cross-sectional images of the body and the use of contrast agents.

[0361] It is possible to test for an increase in survival and life expectancy. To determine the effectiveness of treatment with the IAC of the present invention, a Kaplan-Meier survival analysis can be performed, particularly in a cohort study comparing untreated patients and / or patients treated with standard radioligand therapy (without immune-inducing portion) with patients treated with the IAC according to the present invention.

[0362] To visualize and compare survival (or event-free) probabilities over time using the Kaplan-Meier method, survival data is collected and analyzed using statistical software, such as SPSS, R, or SAS.

[0363] Patient groups (e.g., untreated patients; patients treated with standard radioligand therapy; patients treated with IAC according to the present invention) are monitored for survival, and survival data are entered into selected statistical software. Kaplan-Meier curves are plotted with time on the x-axis and survival probability on the y-axis, with different patient groups represented by different curves. The median survival time, which is the point at which the curve intersects the 50% survival line, is visualized. A log-rank test is performed to compare survival curves between groups, and statistical significance is determined using the p-value generated by the log-rank test (typically, p < 0.05 is considered significant). Those skilled in the art will also interpret the results based on shape, rate of decline, and potential differences between survival curves, as well as significant differences in median survival time and between groups.

[0364] example Example 1 Figures 4A and 4B show the results of the first cure test, with Figure 4A specifically showing the antibody response and Figure 4B showing the total number of CD8+ T cells after treatment. The patient was diagnosed with extensive cutaneous metastases (over 10 cm in diameter) from melanoma and was initially injected intratumor with 100 μg of avidin in 1 mL of PBS buffer. Subsequently, 5 μg of COV-19 spike biotin (biotinylated recombinant SARS-CoV-2 spike His-tag) in 1 mL of PBS buffer was administered. Blood parameters were determined at corresponding intervals.

[0365] Mechanistically, avidin is offered as an intratumoral targeting agent—avidin is large and cannot diffuse. Therefore, avidin is suitable for targeting (in this case, locally).

[0366] The results shown in Figures 4A and 4B clearly demonstrate a decrease in COVID-19 antibodies and total CD8 T cell yield in the patients' blood after 2 hours, indicating a sink effect.

[0367] This suggests that corresponding immune cells migrate into the tumor tissue, enabling the active transport of immune cells into the tumor. Furthermore, the enhanced immune response is induced over a period of 7 days.

[0368] Therefore, the IAC therapy of the present invention will also provide an immune attractant for immune cells to target each tumor cell.

[0369] The immune-inducing compound (IAC) according to the present invention is therefore suitable for treating the target cancer disease. The target structure binding moiety can bind to the target structure of the target cell, and the presence and / or overexpression of the target structure indicates the target cancer disease. Thus, the immune-inducing moiety of the IAC of the present invention can attract immune cells to the target cell.

[0370] Example 2: Preclinical trials The concept of preclinical trials using the immunoinducing compounds according to the present invention can be established by those skilled in the art. In particular, the methods and protocols described by Muller et al. (Cristina Muller, Maria De Prado Leal, Marco D. Dominietto, Christoph A. Umbricht, Sairos Safai, Rosalind L. Perrin, Martina Eglo, Peter Bernhardt, Nicholas P. van der Meulen, Damien C. Weber, Roger Schibli and Antony J. Lomax, “Combination of Proton Therapy and Radionuclide” in Pharmaceutics 2019, 11, 450; doi:10.3390 / pharmaceutics 11090450) can be applied with their respective modifications to study the application of therapies using the immunoinducing compositions according to the present invention.

[0371] Accordingly, preclinical trials can be set up in the following groups using the corresponding IAC of the present invention.

[0372] As an exemplary IAC according to the present invention, affilin-RBD-HEAD-DOTA may be tested in HER-2 positive mice such as SKBR3 or BT474. [Table 4]

[0373] The treatment studies will be conducted using mice that are either COVID-19 vaccinated (Covid19(+)) or COVID-19 naive (Covid19(-)), and treated with one of the Lu therapeutics listed in Table 4 above. The dose to the tumor will be selected based on the authors' experience in TRT. Monotherapy with Lu-HER2-affylin in either therapeutic mode aims to delay tumor growth, but not to completely eradicate tumor xenografts, in order to allow determination of the potential additive or synergistic effects of combination compounds. Each experiment will include four groups of mice, treated on day 0 of the study. The first group of mice (currently receiving treatment) will be injected with saline only (groups A / B: control). The next group will be treated with Lu-HER2-affylin. Groups 5 and 6 will be given combination compounds. In all studies, a radioactivity equivalent to 15 MBq will be injected.

[0374] Mice are monitored by measuring body weight and tumor size every other day for 9 weeks. Mice are euthanized when a predefined endpoint (see below) is reached or when the experiment ends on day 63. Relative body weight (RBW) is defined as [BWx / BW0], where BWx is the body weight in grams on a given day x and BW0 is the body weight in grams on day 0. Tumor dimensions are determined by measuring the longest tumor axis (L) and its vertical axis (W) with a digital caliper. Tumor volume (V) is calculated according to the formula [V=0.5×(L×W2)]. Relative tumor volume (RTV) is defined as [TVx / TV0], where TVx is the tumor volume in mm3 on a given day x and TV0 is the tumor volume in mm3 on day 0.

[0375] The endpoint criteria are set according to the size of the mouse strain. In this study, the endpoint criteria are defined as follows: (i) tumor volume > 1000 mm³; (ii) weight loss of 15% or more; (iii) tumor volume of 900 mm³ or more and weight loss of 10% or more; or (iv) signs of anxiety and discomfort.

[0376] The results can be plotted as a graph, as shown in Figure 2 of Muller et al.'s (Cristina Muller, Maria De Prado Leal, Marco D. Dominietto, Christoph A. Umbricht, Sairos Safai, Rosalind L. Perrin, Martina Eglo, Peter Bernhardt, Nicholas P. van der Mulen, Damien C. Weber, Roger Schibli and Antony J. Lomax, “Combination of Proton Therapy and Radionuclide” in Pharmaceutics 2019, 11, 450; doi:10.3390 / pharmaceutics11090450).

[0377] As a result, groups 1 and 2, respectively, preferably provide similar results to the control group (pink), showing that tumor progression and relative tumor volume rapidly increased from about 1 to 5 RTV within about 14 days without treatment.

[0378] Groups 3, 4, and 6 were mice treated with 177Lu-Forate, respectively (TRT; Muller et al. (Cristina Muller, Maria De Prado Leal, Marco D. Dominietto, Christoph A. Umbricht, Sairos Safai, Rosalind L. Perrin, Martina Eglo, Peter Bernhardt, Nicholas P. van der Meulen, Damien C. Weber, Roger Schibli and Antony J. Lomax, “Combination of Proton Therapy and Radionuclide” in Pharmaceutics) The results are similar to those shown in the blue curve in 2019,11,450;doi:10.3390 / pharmaceutics11090450, and will likely show at least moderate treatment success rates. According to our findings, this slight positive therapeutic effect is due to the HER2-mediated antitumor effect, which has a positive effect on tumor progression and median overall survival in this group, and the RTV will generally increase from 1 to 3-4 within about 28 days.

[0379] Group 5 showed a significant success rate as a result of the combined therapeutic effect of the immune-inducing compounds (IACs) according to the present invention, and the COVID-19 positive group had their respective pre-immune responses, which would be stimulated to antitumor activity in addition to the effects already observed in groups 3, 4, and 6, respectively. According to our findings, this positive therapeutic effect is significant compared to groups 3, 4, and / or 6, respectively, and results in a positive effect on tumor progression and median overall survival in these groups, with RTV generally remaining in the range of 1 to 2 within approximately 20 to 60 days, depending on the overall survival of the mice.

[0380] Sequence List The sequences described in this application are shown below and in the attached sequence listing file. [Table 5-1] [Table 5-2] [Table 5-3] [Table 5-4] [Table 5-5] [Table 5-6] [Table 5-7] [Table 5-8] [Table 5-9]

[0381] Embodiment Embodiment 1: An immunoassay compound (IAC) (1) suitable for treating a target cancer disease, It comprises at least one target structure binding portion (2) and at least one immune-inducing portion (3), The target structure binding portion (2) can bind to the target structure (4) of the target cell (5). The presence and / or overexpression of the target structure (4) indicates the target cancer disease, The immune-attracting portion (3) is an immune-attracting compound (IAC) (1) that can attract immune cells (7) to the target cells (5).

[0382] Embodiment 2: The immunoinducing compound (IAC) (1) according to Embodiment 1, wherein the compound is suitable for treating a target cancer disease.

[0383] Embodiment 3: An immunoinducing compound (IAC) (1) according to Embodiment 1 or 2, which can induce an immune response, particularly a cellular immune response, to the immune inducing portion (3).

[0384] Embodiment 4: An immunoinducing compound (IAC) (1) according to any one of Embodiments 1 to 3, which can induce an immune response against an immune inducing portion (3) or corresponding cells in a subject.

[0385] Embodiment 5: An immunoinducing compound (IAC) (1) according to any one of Embodiments 1 to 4, which can utilize the existing immune cell recognition of an epitope to increase immune cell volume and / or cell death activity in and / or in the target cell environment.

[0386] Embodiment 6: The immune inducer portion (3) comprises an antigen, preferably the antigen is an immune inducer compound (IAC) (1) according to any one of Embodiments 1 to 5, which is known in the target immune system.

[0387] Embodiment 7: An immune inducer compound (IAC) (1) according to any one of Embodiments 1 to 6, wherein the immune inducer portion (3) comprises an antigen expressed by target tumor cells or cells in the tumor cell environment.

[0388] Embodiment 8: An immunoinducing compound (IAC) (1) according to any one of Embodiments 1 to 7, wherein the tumor-specific antigen may be selected to be an antigen specific to a cancer disease different from the cancer disease of interest.

[0389] Embodiment 9: An immunoinducing compound (IAC) (1) according to any one of Embodiments 1 to 8, wherein the antigen is a tumor-specific antigen (TSA).

[0390] Embodiment 10: An immune inducer compound (IAC) (1) according to any one of Embodiments 1 to 9, wherein the immune inducer portion (3) comprises an antigen not expressed by target tumor cells or cells in the tumor cell environment.

[0391] Embodiment 11: An immune inducer compound (IAC) (1) according to any one of Embodiments 1 to 10, wherein the immune inducer portion (3) comprises an antigen selected from pathogen-derived antigens, tumor immune microenvironment (TIME)-related antigens, tumor-associated antigens (TAAs), tumor-specific antigens, tumor germline antigens, neoantigens, artificial immunostimulatory antigens, or some and / or combinations thereof.

[0392] Embodiment 12: An immunoinducing compound (IAC) (1) according to any one of Embodiments 1 to 11, wherein TAA is an antigen associated with the target cancer disease.

[0393] Embodiment 13: An immunoinducing compound (IAC) (1) according to any one of Embodiments 1 to 12, wherein the TAA is an antigen associated with a cancer disease different from the target cancer disease.

[0394] Embodiment 14: An immunoinducing compound (IAC) (1) according to any one of Embodiments 1 to 13, wherein the antigen is a tumor germline antigen, more preferably a tumor germline antigen specific to the target cancer disease.

[0395] Embodiment 15: An immunoinducing compound (IAC) (1) according to any one of Embodiments 1 to 14, wherein the tumor-specific antigen is an antigen specific to the target cancer disease.

[0396] Embodiment 16: An immunoinducing compound (IAC) (1) according to any one of Embodiments 1 to 15, wherein the tumor-specific antigen is an antigen specific to a cancer disease different from the target cancer disease.

[0397] Embodiment 17: An immunoinducing compound (IAC) (1) according to any one of Embodiments 1 to 16, wherein the tumor germline antigen is an antigen specific to the target cancer disease.

[0398] Embodiment 18: An immunoinducing compound (IAC) (1) according to any one of Embodiments 1 to 17, wherein the tumor germline antigen is specific to a second cancer disease different from the target cancer disease.

[0399] Embodiment 19: An immune inducer compound (IAC) (1) according to any one of Embodiments 1 to 18, wherein the immune inducer portion (3) comprises an antigen, the antigen being a vaccine antigen and / or a vaccine-associated antigen.

[0400] Embodiment 20: An immune inducer compound (IAC) (1) according to any one of Embodiments 1 to 19, wherein the immune inducer portion (3) comprises an antigen, and the antigen is an exogenous antigen.

[0401] Embodiment 21: An immunoinducing compound (IAC) (1) according to any one of Embodiments 1 to 20, wherein the immunoinducing portion (3) comprises an antigen, and the antigen is an immunodominant antigen.

[0402] Embodiment 22: An immunoinducing compound (IAC) (1) according to any one of Embodiments 1 to 21, wherein the immunoinducing portion (3) comprises an exogenous antigen selected from a virus and / or bacterial antigen or a part thereof.

[0403] Embodiment 23: An immunoinducing compound (IAC) (1) according to any one of Embodiments 1 to 22, wherein the immune inducing portion (3) comprises an antigen, and the antigen comprises a B or T cell epitope.

[0404] Embodiment 24: An immunoinducing compound (IAC) (1) according to any one of Embodiments 1 to 23, wherein the immunoinducing portion (3) can induce an antibody and / or a TH1 and / or TH2 immune response.

[0405] Embodiment 25: An immune inducer compound (IAC) (1) according to any one of Embodiments 1 to 24, wherein the immune inducer portion (3) comprises an antigen, the antigen being a viral antigen of a vaccine having a vaccination rate of at least 50%, preferably at least 75%, more preferably at least 90%, and even more preferably at least 95%.

[0406] Embodiment 26: An immune inducer compound (IAC) (1) according to any one of Embodiments 1 to 25, wherein the immune inducer portion (3) comprises an antigen, the antigen being a viral antigen of a vaccine having an immunization rate of at least 50%, preferably at least 75%, more preferably at least 90%, and even more preferably at least 95%.

[0407] Embodiment 27: An immune inducer compound (IAC) (1) according to any one of Embodiments 1 to 26, wherein the immune inducer portion (3) comprises an antigen selected from or derived from measles, mumps, rubella (MMR) vaccine, diphtheria, tetanus, polio (DTP) vaccine, Haemophilus influenzae type b (Hib) vaccine, pneumococcal vaccine, hepatitis B vaccine, and SARS-CoV-19 vaccine, or a portion thereof.

[0408] Embodiment 28: An immunoinducing compound (IAC) (1) according to any one of Embodiments 1 to 27, wherein the immune inducing portion (3) comprises an antigen selected from the group including natural substances, antibodies, affilins, peptides, proteins, carbohydrates, lipids, nucleic acids, synthetic compounds or toxoids, or combinations thereof.

[0409] Embodiment 29: An immunoinducing compound (IAC) (1) according to any one of Embodiments 1 to 28, wherein the immune inducing portion (3) comprises an antigen, and the antigen is a synthetic compound selected from the group comprising small molecules.

[0410] Embodiment 30: An immune inducer compound (IAC) (1) according to any one of Embodiments 1 to 29, wherein the immune inducer portion (3) comprises an antigen, and the antigen is a nucleic acid selected from the group including DNA, RNA, mRNA, rRNA, tRNA, miRNA, siRNA, snRNA, piRNA, and lncRNA.

[0411] Embodiment 31: An immunoinducing compound (IAC) (1) according to any one of Embodiments 1 to 30, wherein the immune inducing portion (3) comprises a nucleic acid molecule encoding an amino acid sequence containing the SARS-CoV-2 S protein and / or an immunogenic variant thereof, or an immunogenic fragment of the SARS-CoV-2 S protein or its immunogenic variant.

[0412] Embodiment 31: An immunoinducing compound (IAC) (1) according to any one of Embodiments 1 to 30, wherein the immune inducing portion (3) comprises nucleic acid molecules packed in lipid nanoparticles.

[0413] Embodiment 32: An immune inducer compound (IAC) (1) according to any one of Embodiments 1 to 31, wherein the immune inducer moiety comprises an immunogenic fragment of the SARS-CoV-2 S protein, most preferably in the form of an affilin, the S1 subunit of the SARS-CoV-2 S protein, or the receptor-binding domain (RBD) of the SARS-CoV-2 S protein, preferably the S1 subunit of the SARS-CoV-2 S protein, or a portion thereof.

[0414] Embodiment 32: An immunoinducing compound (IAC)(1) according to any one of Embodiments 1 to 31, wherein the immune inducing portion (3) comprises an amino acid sequence containing the SARS-CoV-2 S protein, an immunogenic mutant thereof, or an immunogenic fragment of the SARS-CoV-2 S protein or its immunogenic mutant, and is encoded by a coding sequence that is codon-optimized and / or has an increased G / C content compared to the wild-type coding sequence, wherein the codon optimization and / or the increased G / C content preferably does not alter the sequence of the encoded amino acid sequence.

[0415] Embodiment 33: The immune-inducing portion (3) is Preferably, a nucleic acid molecule, particularly an RNA molecule, encoding the SARS-CoV-2 S protein, its immunogenic variant, or an immunogenic fragment of the SARS-CoV-2 S protein or its immunogenic variant, comprising the nucleotide sequence of nucleotides 979-1584 of SEQ ID NO: 1, 2, or 3, a nucleotide sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the nucleotide sequence of nucleotides 979-1584 of SEQ ID NO: 1, 2, or 3, or a fragment of a nucleotide sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the nucleotide sequence of nucleotides 979-1584 of SEQ ID NO: 1, 2, or 3; and / or Preferably, an immunogenic fragment of the SARS-CoV-2 S protein, an immunogenic variant thereof, or an immunogenic fragment of the SARS-CoV-2 S protein or an immunogenic variant thereof, comprising the amino acid sequence of amino acids 327-528 of SEQ ID NO: 4, an amino acid sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the amino acid sequence of amino acids 327-528 of SEQ ID NO: 4, or an immunogenic fragment of the amino acid sequence of amino acids 327-528 of SEQ ID NO: 4, having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the amino acid sequence of amino acids 327-528 of SEQ ID NO: 4; and / or Preferably, the SARS-CoV-2 S protein RBD, its immunogenic variant, or immunogenic variant of the SARS-CoV-2 S protein, comprising the amino acid sequence of SEQ ID NO: 8, an amino acid sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the amino acid sequence of SEQ ID NO: 8, or an immunogenic fragment of an amino acid sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the amino acid sequence of SEQ ID NO: 8. including, An immunoinducing compound (IAC) (1) according to any one of Embodiments 1 to 32.

[0416] Embodiment 34: The immune induction portion (3) is A nucleic acid molecule, particularly an RNA molecule, encoding the SARS-CoV-2 S protein, its immunogenic variant, or an immunogenic fragment of the SARS-CoV-2 S protein or its immunogenic variant, comprising the nucleotide sequence 49-2055 of SEQ ID NO: 1, 2, or 3, a nucleotide sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the nucleotide sequence 49-2055 of SEQ ID NO: 1, 2, or 3, or a fragment of a nucleotide sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the nucleotide sequence 49-2055 of SEQ ID NO: 1, 2, or 3; and / or Preferably, an immunogenic fragment of the SARS-CoV-2 S protein, an immunogenic variant thereof, or an immunogenic fragment of the SARS-CoV-2 S protein or an immunogenic variant thereof, comprising the amino acid sequence of amino acids 17-685 of SEQ ID NO: 4, an amino acid sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the amino acid sequence of amino acids 17-685 of SEQ ID NO: 4, or an immunogenic fragment of the amino acid sequence of amino acids 17-685 of SEQ ID NO: 4, having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the amino acid sequence of amino acids 17-685 of SEQ ID NO: 4; and / or Preferably, the SARS-CoV-2 S protein RBD, its immunogenic variant, or immunogenic variant of the SARS-CoV-2 S protein, comprising the amino acid sequence of SEQ ID NO: 8, an amino acid sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the amino acid sequence of SEQ ID NO: 8, or an immunogenic fragment of an amino acid sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the amino acid sequence of SEQ ID NO: 8. including, An immunoinducing compound (IAC) (1) according to any one of Embodiments 1 to 33.

[0417] Embodiment 35: The immune-inducing portion (3) is A nucleic acid molecule, particularly an RNA molecule, encoding the SARS-CoV-2 S protein, its immunogenic variant, or an immunogenic fragment of the SARS-CoV-2 S protein or its immunogenic variant, comprising the nucleotide sequence 49-3819 of SEQ ID NO: 1, 2, or 3, a nucleotide sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the nucleotide sequence 49-3819 of SEQ ID NO: 1, 2, or 3, or a fragment of a nucleotide sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the nucleotide sequence 49-3819 of SEQ ID NO: 2, 8, or 9 or the nucleotide sequence 49-3819 of SEQ ID NO: 1, 2, or 3; and / or Preferably, an immunogenic fragment of the SARS-CoV-2 S protein, an immunogenic variant thereof, or an immunogenic fragment of the SARS-CoV-2 S protein or its immunogenic variant, comprising the amino acid sequence of amino acids 17-1273 of SEQ ID NO: 4 or 5, an amino acid sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the amino acid sequence of amino acids 17-1273 of SEQ ID NO: 4 or 5, or an immunogenic fragment of the amino acid sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the amino acid sequence of amino acids 17-1273 of SEQ ID NO: 4 or 5; and / or Preferably, the SARS-CoV-2 S protein RBD, its immunogenic variant, or immunogenic variant of the SARS-CoV-2 S protein, comprising the amino acid sequence of SEQ ID NO: 8, an amino acid sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the amino acid sequence of SEQ ID NO: 8, or an immunogenic fragment of an amino acid sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the amino acid sequence of SEQ ID NO: 8. including, An immunoinducing compound (IAC) (1) according to any one of Embodiments 1 to 34.

[0418] Embodiment 36: An immunoinducing compound (IAC) (1) according to any one of Embodiments 1 to 35, wherein the immune inducing portion (3) comprises an amino acid sequence containing the SARS-CoV-2 S protein, an immunogenic variant thereof, or an immunogenic fragment of the SARS-CoV-2 S protein or its immunogenic variant, and comprises a secretory signaling peptide.

[0419] Embodiment 37: The immunoinducing compound (IAC)(1) according to Embodiment 36, wherein the secretory signaling peptide is preferably fused at its N-terminus to the SARS-CoV-2 S protein, an immunogenic mutant thereof, or an immunogenic fragment of the SARS-CoV-2 S protein or an immunogenic mutant thereof.

[0420] Embodiment 38: The immune induction portion (3) is RNA molecules encoding a secretory signal peptide, comprising the nucleotide sequence of nucleotides 1-48 of SEQ ID NO: 1, 2, or 3; a nucleotide sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the nucleotide sequence of nucleotides 1-48 of SEQ ID NO: 1, 2, or 3; or a fragment of a nucleotide sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the nucleotide sequence of nucleotides 1-48 of SEQ ID NO: 1, 2, or 3; and / or Secretory signal peptides containing a functional fragment of the amino acid sequence of amino acids 1-16 of SEQ ID NO: 4, an amino acid sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the amino acid sequence of amino acids 1-16 of SEQ ID NO: 4, or an amino acid sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the amino acid sequence of amino acids 1-16 of SEQ ID NO: 4. An immunoinducing compound (IAC) (1) according to any one of embodiments 1 to 37, including the above.

[0421] Embodiment 39: The immune induction portion (3) is The RNA encoding the SARS-CoV-2 S protein, its immunogenic variant, or an immunogenic fragment of the SARS-CoV-2 S protein or its immunogenic variant comprises the nucleotide sequence of SEQ ID NO: 6, a nucleotide sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the nucleotide sequence of SEQ ID NO: 6, or a fragment of a nucleotide sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the nucleotide sequence of SEQ ID NO: 6; and / or SARS-CoV-2 S protein, its immunogenic variant, or immunogenic fragments of the SARS-CoV-2 S protein or its immunogenic variant are defined as the amino acid sequence of SEQ ID NO: 7, an amino acid sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the amino acid sequence of SEQ ID NO: 7, or an immunogenic fragment of an amino acid sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the amino acid sequence of SEQ ID NO: 7. An immunoinducing compound (IAC) (1) according to any one of embodiments 1 to 38, including the above.

[0422] Embodiment 40: An immunoinducing compound (IAC) (1) according to any one of Embodiments 1 to 39, wherein the immunoinducing portion (3) comprises a peptide having a length of at least 8 amino acids, preferably at least 10 amino acids, and more preferably at least 15 amino acids.

[0423] Embodiment 41: An immunoinducing compound (IAC) (1) according to any one of Embodiments 1 to 40, wherein the immunoinducing portion (3) comprises a peptide having a length of up to 200 amino acids, preferably up to 150 amino acids, more preferably up to 100 amino acids, even more preferably up to 65 amino acids, even more preferably up to 60 amino acids, and even more preferably up to 25 amino acids.

[0424] Embodiment 42: An immune inducer compound (IAC) (1) according to any one of Embodiments 1 to 41, wherein the immune inducer portion (3) comprises a subunit, recombinant or conjugated vaccine, or an antigen derived therefrom or a part thereof.

[0425] Embodiment 43: An immunoinducing compound (IAC) (1) according to any one of Embodiments 1 to 42, wherein the immune inducing portion (3) is a toxoid or comprises an antigen or a part thereof derived from a toxoid, or the immune inducing portion comprises a toxoid and an antigen expressed by target cells.

[0426] Embodiment 44: An immune inducer compound (IAC) (1) according to any one of Embodiments 1 to 43, wherein the immune inducer portion (3) comprises a protein derived from the SARS-CoV-19 virus, in particular the SARS-CoV-19 spike protein, or an antigen which is a part thereof.

[0427] Embodiment 45: An immune inducer compound (IAC) (1) according to any one of Embodiments 1 to 44, wherein the immune inducer portion (3) preferably includes the amino acid sequence of SEQ ID NO: 8, an amino acid sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the amino acid sequence of SEQ ID NO: 8, or an immunogenic fragment of the amino acid sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the amino acid sequence of SEQ ID NO: 8 or the amino acid sequence of SEQ ID NO: 8, and includes an antigen which is an immunogenic fragment of the SARS-CoV-19 virus-derived RBD pr...

Claims

1. Immuno-inducing compounds (IACs) suitable for treating the target cancer disease, including the following: (1) At least one target structure binding site (2) and at least one immune-inducing site (3), however, The target structure binding portion (2) can bind to the target structure (4) of the target cell (5). The presence and / or overexpression of the target structure (4) indicates the target cancer disease, and The immune-attracting portion (3) can attract immune cells (7) to the target cells (5), and preferably the immune-attracting portion (3) contains a protein derived from the SARS-CoV-19 virus, particularly the SARS-CoV-19 spike protein, or an antigen which is a part thereof, and The IAC (1) optionally includes a chelating agent (91) linked to the target structure binding portion (2) and / or the immune induction portion (3).

2. Immuno-inducing compounds (IACs) suitable for treating the target cancer disease, including the following (1): At least one target structure binding site (2) and at least one immune-inducing site (3), however, The target structure binding portion (2) can bind to the target structure (4) of the target cell (5). The presence and / or overexpression of the target structure (4) indicates the target cancer disease, the target structure (4) is Her2 Neu, and preferably the target structure binding portion (2) comprises Her2 Neu-binding affilin (2, 21), and The immune-attracting portion (3) can attract immune cells (7) to the target cells (5), The immune-inducing portion (3) includes a protein derived from the SARS-CoV-19 virus, particularly the SARS-CoV-19 spike protein, or an antigen which is a part thereof, and The IAC (1) optionally includes a chelating agent (91) linked to the target structure binding portion (2) and / or the immune induction portion (3).

3. IAC contains a chelating agent (91), (i) The chelating agent is linked to the target structure binding portion (2), (ii) The chelating agent can bind to the radioactive portion (9), preferably the radioactive portion (9) is a radionuclide (9) and / or (iii) The target structure binding portion (2) is linked to the immune induction portion (3) via the linker portion (8), and the IAC, in particular the linker (8), comprises the chelating agent (91) and / or (iv) The target structure binding portion (2) is linked to the immune induction portion (3) via a linker portion (8), wherein the IAC, in particular the linker (8), comprises the chelating agent (91) and complexes the radioactive portion (9), wherein the radioactive portion is optionally gallium-68 (Ga-68) and / or lutetium-177 (Lu-177) or actinium-225 (Ac-225) or yttrium-90 (Y-90) or any other radiodiagnostic or radiotherapeutic isotope. An immunoassay compound (IAC) according to claim 1 or 2 (1).

4. An immune inducer compound (IAC) (1) according to any one of claims 1 to 3, wherein the immune inducer moiety comprises an immunogenic fragment of the SARS-CoV-2 S protein, most preferably in the form of an affilin, the S1 subunit of the SARS-CoV-2 S protein, or the receptor-binding domain (RBD) of the SARS-CoV-2 S protein, preferably the S1 subunit of the SARS-CoV-2 S protein, or a portion thereof.

5. The immune inducer compound (IAC) (1) according to any one of claims 1 to 4, wherein the immune inducer portion (3) comprises an amino acid sequence comprising a SARS-CoV-2 S protein encoded by a coding sequence in which the codon optimization and / or the G / C content is increased compared to the wild-type coding sequence, an immunogenic mutant thereof, or an immunogenic fragment of the SARS-CoV-2 S protein or its immunogenic mutant, wherein the codon optimization and / or the increase in G / C content preferably does not alter the sequence of the encoded amino acid sequence.

6. The immune inducer compound (IAC) (1) according to any one of claims 1 to 5, wherein the immune inducer portion (3) comprises an amino acid sequence containing SARS-CoV-2 S protein, an immunogenic mutant thereof, or an immunogenic fragment of the SARS-CoV-2 S protein or its immunogenic mutant, and the amino acid sequence comprises a secretory signaling peptide.

7. The immunoinducing compound (IAC) (1) according to claim 6, wherein the secretory signal peptide is preferably fused at its N-terminus to the SARS-CoV-2 S protein, an immunogenic mutant thereof, or an immunogenic fragment of the SARS-CoV-2 S protein or its immunogenic mutant.

8. An immune inducer compound (IAC) (1) according to any one of claims 1 to 7, wherein the immune inducer portion (3) preferably comprises an amino acid sequence of SEQ ID NO: 8, an amino acid sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the amino acid sequence of SEQ ID NO: 8, or an immunogenic fragment of an amino acid sequence having at least 99%, at least 98%, at least 97%, at least 96%, at least 95%, at least 90%, at least 85%, or at least 80% identity with the amino acid sequence of SEQ ID NO: 8 or the amino acid sequence of SEQ ID NO: 8, and comprises an antigen which is an immunogenic fragment of the RBD protein derived from the SARS-CoV-19 virus, preferably the RBD of the SARS-CoV-2 S protein, an immunogenic variant thereof, or an immunogenic fragment of the RBD of the SARS-CoV-2 S protein or an immunogenic variant thereof.

9. An immunoassay compound (IAC) (1) (i) One or more, preferably two, identical immune-inducing moieties (3), or (ii) More than one, preferably two or more, different immune-inducing moieties (3) An immunoassay compound (IAC) (1) according to any one of claims 1 to 8, comprising:

10. (i) The target cell (5) is a cancer cell, and the disease is a cancerous disease. (ii) The cancerous disease is breast cancer, and / or (iii) The target structure (4) is expressed by the target cell (5), An immunoassay compound (IAC) according to any one of claims 1 to 9 (1).

11. An immunoassay compound (IAC) according to any one of claims 1 to 11, for use as a pharmaceutical (1).

12. An immunoassay compound (IAC) according to any one of claims 1 to 11 (1) for use in treating cancer.

13. An immunoinducing compound (IAC) (1) according to any one of claims 1 to 11 for use in a method for treating cancer in a subject, wherein the method comprises step a) of applying the immunoinducing compound (IAC) (1) according to any one of claims 1 to 10 to the subject, the subject being a mammal, preferably a human.

14. An immunoinducing compound (IAC) (1) according to any one of claims 1 to 11 for use in the method of claim 14, the method further comprising the step of determining the initial immune state of a subject to at least one antigen, preferably the step of determining the initial state of the immune response after vaccination of the subject.

15. An immune inducer compound (IAC) (1) according to any one of claims 1 to 11 for use in the method according to claim 14 or 15, the method further comprising the step of determining the responsive immune state of a subject to at least one antigen, preferably the step of determining the response state of the immune response after treatment with the IAC according to any one of claims 1 to 11.

16. An immunoinducing compound (IAC) (1) according to any one of claims 1 to 11, for use in the method of any one of claims 14 to 16, further comprising the step of vaccinating a subject with a vaccine relating to any antigen contained in at least one immunoinducing portion (3) of an immunoinducing compound (IAC) (1) administered in step a).

17. An immune inducer compound (IAC) (1) according to any one of claims 1 to 11 for use in the method according to any one of claims 14 to 17, the method further comprising a second step of determining the immune status of a subject to at least one antigen, preferably a second step of determining the state of the immune response of the subject after vaccination in the step according to claim 17.

18. An immune inducer compound (IAC) (1) according to any one of claims 1 to 11, for use in the method according to any one of claims 14 to 18, further comprising the step of additionally vaccinating a target with a vaccine related to an antigen contained in at least one immune inducer portion (3) of the immune inducer compound (IAC) (1) administered in step a).

19. It comprises at least one target structure binding portion (2) and at least one immune-inducing portion (3), The target structure binding portion (2) is connected to the immune induction portion (3) via the linker portion (8), The target structure binding portion (2) is It can bind to the target structure (4) of the target cell (5), The presence and / or overexpression of the target structure (4) indicates the target cancer disease, The immune-attracting portion (3) can attract immune cells (7) to the target cells (5), The target structure binding portion (2) is Her2 Neu-binding affilin (2, 21), The immune-inducing portion (3) is the SARS-CoV-19 spike protein, particularly the RBD of the SARS-CoV-19 spike protein, or a part thereof. IAC (1) comprises a chelating agent (91), the chelating agent (91) is linked to the target structure binding portion (2), The chelating agent (91) complexes the radioactive moiety (9), preferably gallium-68 (Ga-68) and / or lutetium-177 (Lu-177), or actinium-225 (Ac-225), or yttrium-90 (Y-90), or any other radiodiagnostic or radiotherapeutic isotope. An immunoassay compound (IAC) (1) suitable for treating the target cancer disease, preferably according to any one of claims 1 to 19.

20. A pharmaceutical composition comprising an immunoassay compound (IAC) (1) according to any one of claims 1 to 20, and optionally a suitable carrier.

21. A pharmaceutical composition according to claim 21 for use in the treatment, in vivo diagnosis and / or prevention of cancer.

22. Step a) to prepare at least one target structure binding moiety (2) precursor molecule. Step b) to prepare at least one immune-inducing moiety (3) precursor molecule. Step c) Linking the at least one target structure binding portion (2) precursor molecule to the at least one immune attraction portion (3) precursor molecule. A method for producing an immunoassay compound (IAC) (1) according to any one of claims 1 to 20, comprising:

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