• 专利附录
  • 专利说明
  • 权利要求
  • 类似技术
  • 同族专利
  • 引用文献
  • 法律状态
  • 优先权
    • 专利摘要:
    Methods and pharmaceutical compositions for enhancing endogenous immune response-mediated clearance of pathogenic cell populations in host animals are provided wherein the pathogenic cells preferentially express, uniquely express or overexpress binding sites for specific ligands. . The present invention consists in administering a conjugated ligand to an immunogen capable of activating toll-like receptors to a host animal including a pathogenic cell population. At least one other therapeutic factor may be administered, wherein the therapeutic factor is a compound capable of promoting an endogenous immune response, and the compound does not bind to the ligand-immunogen conjugate.
    公开号:KR20040053136A
    申请号:KR10-2004-7004233
    申请日:2002-09-26
    公开日:2004-06-23
    发明作者:필립스튜어트 로우；잉주안 류
    申请人:펄듀 리서치 파운데이션；
    IPC主号:
    专利说明:

    METHOD OF TREATMENT USING LIGAND-IMMUNOGEN CONJUGATE}
    [2] The mammalian immune system provides a means for recognizing and eliminating tumor cells, other pathogenic cells, and invading foreign pathogens. Although the immune system generally provides a strong line of defense, in many cases cancer cells, other pathogenic cells, or infectious agents proliferate away from the host immune response or persist in concomitant host pathogenicity. Chemotherapy and radiotherapy have been developed to eliminate cloned neoplasms. However, side effects occur because currently available chemotherapeutic agents and radiotherapy not only destroy cancer cells but also affect normal host cells, such as hematopoietic cells. In addition, chemotherapeutic agents have limited effectiveness when host drug resistance develops.
    [3] In addition, foreign pathogens can proliferate in the host by avoiding a competent immune response or when the host immune system is weakened by drug therapy or other health problems. Although many therapeutic compounds have been developed, pathogens develop resistance to these therapeutic agents. Due to the ability of cancer cells and infectious agents to develop resistance to therapeutic drugs and the side effects of currently available anticancer drugs, there is a need for the development of novel therapies specific to the pathogenic cell population with less host toxicity.
    [4] The researchers developed a therapeutic protocol that specifically targets cytotoxic compounds to these cells to destroy cancer cells. These protocols use toxins conjugated to ligands that bind to receptors that are unique to cancer cells or overexpressed by cancer cells in order to minimize normal cell delivery of toxins. Using this approach, specific immunotoxins have been developed that consist of antibodies directed to specific receptors of pathogenic cells, which antibodies are linked to toxins such as lysine, Pseudomonas exotoxin, diphtheria toxin, tumor necrosis factor. These immunotoxins target tumor cells carrying receptors specifically recognized by antibodies (Olsnes, S., Immunol. Today , 10, pp . 291-295, 1989; Melby, EL, Cancer Res. , 53 (8), pp. 1755-1760, 1993; Better, MD, PCT Publication No. WO 91/07418, published May 30, 1991).
    [5] Another way of selectively targeting cancer cells or foreign pathogens in the host is to enhance host immune responses against pathogenic cells, thereby avoiding the administration of compounds with potential for independent host toxicity. One strategy of immunotherapy is to bind antibodies, eg, genetically engineered complex antibodies, to the tumor cell surface, presenting constant regions of the antibody at the cell surface, and inducing tumor cell death by various immune system mediated processes (De Vita). , VT, Biologic Therapy of Cancer , 2d ed.Philadelphia, Lippincott, 1995; Soulillou, JP, US Patent No. 5,672,486). However, it is difficult to define tumor-specific antigens in this manner. Another way that relies on host immune competency is to target anti-T cell receptor antibodies or anti-Fc receptor antibodies to the tumor cell surface to promote direct binding of immune cells to tumors (Kranz, DM, US Patent No. 5,547,668). In addition, a vaccine-based approach has been proposed, which is based on vaccines containing antigens fused to cytokines, which alter the immunogenicity of vaccine antigens and promote immune responses against pathogenic factors (Pillai, S., PCT Publication No. WO 91/11146, published Feb. 7, 1991). This method is based on indirect regulation of the reported immune response. Another way to kill unwanted cell populations is to use IL-2, or Fab fragments of anti-thymocyte globulin linked to the antigen, to remove unwanted T cells; However, the reported experimental data show that the method removes only about 50% of the targeted cell population and induces nonspecific cell death in vivo (ie, kills 50% of peripheral blood lymphocytes and not T cells (Pouletty, P., PCT Publication No. WO 97/37690, October 16, 1997). Thus, there is an urgent need for therapies that can treat disease states characterized by the presence of a pathogenic cell population in a diseased host.
    [6] Immunity can be divided into innate or adaptive immunity, which is mediated by T and B cells that exhibit specificity and memory. Innate immune responses are immediate responses that can be mediated by phagocytes, natural killer cells, T cells, other cells, and the complement system. Phagocytes, such as monocytes / macrophages and dendritic cells (antigen-presenting cells), kill and degrade infectious agents or foreign antigens, and present the components of ingested pathogens in naïve T cells as MHC or MHC-like autoantigen Function Naïve T cells recognize foreign antigens as MHC or MHC-like autoantigens and differentiate into T H 1 or T H 2 cells capable of inducing adaptive immunity.
    [7] Innate immunity-mediated immune cells can distinguish between autologous and pathogenic antigens by utilizing surface receptors that recognize motifs conserved in pathogens that do not exist in higher eukaryotes. One receptor of this kind is a toll-like receptor that is conserved between insects and humans and first identified in Drosophila . Toll-like lineage receptors have at least 10 members (TLR 1-10) and are present on the surface of monocytes / macrophages and dendritic cells. Although toll-like receptors are activated through the recognition of conserved motifs in pathogens, whether such activation occurs through direct binding of the conserved motifs with toll-like receptors or through covalent receptors interacting with toll-like receptors. It is not confirmed. Activation of toll-like receptors results in induction of cytokine secretion and innate immune responses, where naïve T cells differentiate into T H 1 or T H 2 cells. Thus, toll-like receptors may provide an important link between innate immune recognition and activation of subsequent adaptive immunity.
    [8] Molecules capable of activating toll-like receptors are components of pathogenic factors, for example, Muramil dipeptide, LPS, lipopeptides, lipoproteins, peptidoglycans, lipoteachoic acid, lipoarabinomannan ), Zymosan (yeast cell wall preparation), or other endogenous molecules of the host, such as fibronectin or heat shock proteins. Toll-like receptors can be activated indirectly through covalent receptors or directly with these molecules that promote cytokine secretion from monocytes / macrophages and dendritic cells and result in an innate immune response.
    [9] The present invention relates to a method for removing a pathogenic cell population from a host by increasing host immune system recognition and response to the pathogenic cell population. Indeed, increasing the antigenicity of cell pathogens enhances endogenous immune response-mediated clearance of pathogenic cell populations. This method avoids or minimizes the use of cytotoxic or antimicrobial therapies. The method consists of administering a ligand-immunogen conjugate, wherein the ligand is capable of specifically binding in vivo to a pathogenic cell population that uniquely expresses, preferentially or overexpresses a ligand binding moiety and Immunogens can activate toll-like receptors and induce innate immune responses in host animals. Immune system mediated elimination of pathogenic cells is accomplished by binding the ligand component in a ligand-immunogen conjugate to a receptor, transporter or other surface-presented protein that is uniquely expressed, overexpressed or preferentially expressed by the pathogenic cell. Surface-presented proteins that are uniquely expressed, overexpressed or preferentially expressed by pathogenic cells are receptors that are absent or trace amounts present in non-pathogenic cells and provide a means of selective removal of pathogenic cells. One or more other therapeutic factors, such as immune system promoters, may be co-administered to the host animal to enhance the therapeutic effect.
    [10] In one embodiment, the invention comprises administering a ligand capable of specifically binding with high affinity in vivo to a cell surface protein that is uniquely expressed, preferentially expressed or overexpressed in a targeted cell population ( The ligand consists of activating toll-like receptors and conjugated to an immunogen capable of inducing innate immunity in the host animal) and co-administering one or more therapeutic factors which are endogenous immune response activators. In a preferred embodiment, the present invention involves administering a ligand-immunogen conjugate composition to a host animal, wherein the ligand is a folic acid or other folate receptor binding ligand (e.g., a folate receptor binding ligand independent of folic acid analogs or folates). to be. Ligands are conjugated by covalent bonds to immunogens, for example. Along with administration of the ligand-immunogen conjugate, one or more other therapeutic factors may be administered that are not able to specifically bind the ligand-immunogen complex but may promote or enhance an endogenous immune response.
    [11] In another embodiment there is provided a method of enhancing endogenous immune response-mediated specific clearance of these cell populations in a host animal comprising a pathogenic cell population, wherein members of the cell population retain available binding sites for ligands. do. The method comprises administering to a host a ligand-immune conjugate composition containing a complex of ligand and an immunogen, wherein the immunogen can activate toll-like receptors and the ligand is a vitamin or derivative or analog thereof. The therapeutic factor is a compound capable of promoting an endogenous immune response, which compound consists of administering to the host at least one additional composition comprising a ligand-immunogen conjugate.
    [12] In a preferred embodiment there is provided a method of enhancing endogenous immune response-mediated specific clearance of a pathogenic cell population in a host animal comprising the pathogenic cell population, wherein said cell population uniquely expresses or preferentially expresses a vitamin receptor. Or overexpress. The method comprises administering to a host a composition containing a ligand covalently linked to an immunogen, wherein the immunogen can activate toll-like receptors, wherein the ligand is a vitamin or derivative or analog thereof.
    [13] In other embodiments, the targeted pathogenic cell population is a cancer cell population. In another embodiment, the targeted cell population is a virus-infected cell. In another embodiment, the targeted cell population is an exogenous organism, bacteria, mycoplasma yeast or fungal population. In another embodiment, the targeted cell population is an activated macrophage population that mediates a disease state. Ligand-immunogen conjugates bind to the surface of tumor cells or pathogenic organisms and “label” cell members of a targeted cell population with an immunogen, inducing an immune-mediated response directed at a labeled cell population. Immunogens are directly recognized by immune cells and pathogenic cells can be killed directly.
    [14] Removal of foreign pathogens, infected or neoplastic endogenous cells or pathogenic cells can be enhanced by administering therapeutic factors that promote endogenous immune responses. In one embodiment, the immunostimulator is an interleukin such as IL-2, IL-12, IL-15, IL-18; IFNs such as IFN-α, IFN-β, IFN-γ; Or GM-CSF. In other embodiments, the immunostimulator is a combination of cytokines such as IL-2, IL-12, IL-15 or an effective combination thereof in combination with IFN-α, IFN-β, IFN-β or GM-CSF , Or any other effective cytokine combination. These identified cytokines promote the T H 1 response, but cytokines that promote the T H 2 response, such as IL-4, IL-10, IL-11, or an effective combination thereof, may also be used. In addition, cytokines that promote the T H 1 response and cytokines that promote the T H 2 response may be used in combination.
    [15] In another embodiment a pharmaceutical composition is presented. Such pharmaceutical compositions consist of a therapeutically effective amount of a ligand-immunogen conjugate (immunogen can activate toll-like receptors, the ligand being a vitamin or a derivative or analog thereof) and a pharmaceutically acceptable carrier.
    [16] In another embodiment, ligand-immunogen conjugates (immunogen can activate toll-like receptors, ligands are vitamins or derivatives or analogs thereof), therapeutic factors (therapeutic factors can promote endogenous immune responses) Compounds, which do not bind ligand-immunogen conjugates), pharmaceutical compositions consisting of pharmaceutically active carriers. In one embodiment, the pharmaceutical composition is an extra-intestinal long-term release dosage form. In other embodiments, the therapeutic factor is interleukin (eg, IL-2, IL-4, IL-10, IL-11, IL-12, IL-15), IFN (eg, IFN-α, IFN-β, IFN -γ), GM-CSF, and an immunostimulating agent consisting of a compound selected from a combination thereof.
    [1] The present invention relates to methods and pharmaceutical compositions effective for treating a disease state characterized by the presence of a pathogenic cell population. More specifically, cell-targeted ligand-immunogen complexes, optionally in combination with immune system promoters, are administered to enhance or redirect the host immune response against pathogenic cells.
    [17] 1 depicts the effect of Muramil dipeptide and Muramil dipeptide-folate therapy on tumor-grafted mice.
    [18] 2 shows the effect of CpG and CpG-folate therapy on tumor-grafted mice.
    [19] A method of treating a host in a disease state mediated by pathogenic cells, or a host infected with a pathogenic organism, is provided. This method enhances immune response-mediated clearance of pathogenic cell populations by labeling pathogenic cells with antigens and results in their recognition and removal by the host immune system. The method employs ligand-immunogen conjugates capable of binding to cancer cells or other pathogenic factors or cells with high affinity. High affinity binding can be inherent in the ligand, which can be modified (strengthened) with the use of chemically modified ligands or with specific chemical bonds between the immunogen and the ligands present as conjugates. The method may also utilize combination therapies using additional therapeutic factors capable of enhancing immune response-mediated clearance of ligand-immunogen conjugates and pathogenic cell populations.
    [20] The methods of the present invention are used to enhance endogenous immune response-mediated clearance of pathogenic cell populations in host animals, including pathogenic cell populations. The present invention can be applied to a variety of pathogenic cell populations causing cancer, inflammation, autoimmune diseases, infectious diseases. Thus, the pathogenic cell population may or may not form a tumorigenic cancer cell population, including benign and malignant tumors. Cancer cell populations may occur spontaneously or by a process such as mutation or somatic mutation in a germline of a host animal, or may be induced by chemicals, viruses or radiation. The present invention can be used to treat cancers such as carcinoma, sarcoma, lymphoma, Hodgkin's disease, melanoma, mesotheliomas, Burkitt's lymphoma, thickening carcinoma, leukemia, myeloma. Cancer cell populations include oral cancer, thyroid cancer, endocrine cancer, skin cancer, gastric cancer, esophageal cancer, laryngeal cancer, pancreatic cancer, colon cancer, bladder cancer, bone cancer, ovarian cancer, cervical cancer, uterine cancer, breast cancer, testicular cancer, prostate cancer, rectal cancer, kidney cancer, liver cancer, lung cancer. Included, but not limited to these.
    [21] The pathogenic cell population may also be an exogenous pathogen or a cell population comprising an exogenous pathogen (eg a virus). The invention can be applied to exogenous pathogens such as bacteria, fungi, viruses, mycoplasmas, parasites. Infectious agents that can be treated with the present invention include bacteria such as gram-negative / gram-positive cocci or rods; DNA viruses such as papilloma virus, parvovirus, adenovirus, herpes virus, vaccinia virus; Pathogenesis in animals, including organisms such as RNA viruses such as arenavirus, coronavirus, rhinovirus, respiratory syncope virus, influenza virus, picornavirus, paramyxovirus, leovirus, retrovirus, rhabdovirus Is any infectious organism known in the art. In particular, antibiotic resistant bacteria, such as antibiotic-resistant Streptococcus species and Staphlococcus species, or bacteria that are sensitive to antibiotics but induce relapses of infections treated with antibiotics and eventually develop into resistant bacteria Corresponds to this. These organisms can be treated with the ligand-immunogen conjugates of the present invention in combination with a dose of antibiotic less than that normally administered to a patient in order to avoid the occurrence of said antibiotic-resistant bacterial strains. The invention can be applied to any fungi, mycoplasma species, parasites, or other infectious organisms that cause disease in animals. Examples of fungi that can be treated with the methods of the invention include fungi-growing or yeast-like fungi, such as ringworm, histoplasmosis, blastomycosis, aspergillosis, It is a fungus that causes diseases such as cryptococcosis, spororotrichosis, coccidioidomycosis, South American yeast (paracoccidio-idomycosis) and candidiasis. The invention vivo tapeworm (tapeworm), blood flukes (fluke), tissue roundworms (roundworm), amoeba, flasks modium (Plasmodium) species, trees Fano dressage (Trypanosoma) species, risyu mania (Leishmania) species, Toxoplasma (Toxoplasma) It can be used to treat parasitic infections, including but not limited to infections caused by the species. In particular, parasites that express folate receptors and bind to folates; However, the existing literature sufficiently reports ligands showing high affinity for infectious organisms. For example, penicillin and cephalosporin, which are known for antibiotic activity and specific binding to bacterial cell wall precursors, can similarly be used as ligands for making ligand-immunogen conjugates used in the present invention. The ligand-immunogen conjugate of the present invention may be directed to a cell population comprising endogenous pathogens, wherein the pathogen-specific antigen is preferentially expressed on the surface of the cell containing the pathogen and acts as a receptor for the ligand. The ligand specifically binds to the antigen.
    [22] Furthermore, the pathogenic cell population may be an activated macrophage population that mediates the disease state. Activated macrophages often overexpress cell surface proteins such as folate receptors. Activated macrophages aggravate disease conditions such as ulcerative colitis, Crohn's disease, rheumatoid arthritis, osteomyelitis, atherosclerosis, graft-versus-host disease, psoriasis, osteoporosis, pseudosarcoma, multiple sclerosis, and other inflammatory and autoimmune diseases May cause The activated macrophage population may be an activated macrophage population infected with pathogens such as Salmonella , Shigella , and Tuberculosis species. Activated macrophages are targeted for elimination as a result of ligand-immunogen conjugate binding to the activated macrophage surface.
    [23] The method of the present invention can be used for human clinical treatment or livestock treatment. Thus, host animals, including pathogenic organism populations and that can be treated with ligand-immunogen conjugates, are human or experimental animals, livestock, pets or wildlife in the case of livestock treatment. The present invention is a human; Laboratory animals such as rodents (eg, mice, rats, hamsters, etc.), rabbits, monkeys, chimpanzees; Pets such as dogs, cats, rabbits; Livestock such as cattle, horses, pigs, sheep, and goats; Can be applied to wildlife such as bears, pandas, lions, tigers, leopards, elephants, zebras, giraffes, gorillas, dolphins, whales.
    [24] Suitably, the ligand-immunogen conjugate is administered extrahost, for example intradermal, subcutaneous, intramuscular, intraperitoneal or intravenous to the host animal. Alternatively, the conjugates may be administered to the host animal by other medically useful procedures, such as oral administration, and suitable therapeutic forms may be employed, including effective amounts and long-term release forms. The methods of the present invention may be used for the surgical removal of tumors, radiotherapy, chemotherapy, or biotherapy (eg, monoclonal antibody therapy, treatment with immunomodulators, adoptive transfer of immune effector cells, treatment with hematopoietic growth factors, Cytokines, vaccinations, and other immunotherapy].
    [25] According to the present invention, the ligand component of the ligand-immunogen conjugate can be selected from various ligands. The ligand should be able to specifically remove the pathogenic cell population from the host animal that preferentially expresses the ligand receptor used for ligand binding in the pathogenic cell. Available ligands include folic acid, folic acid analogs and other folate receptor-binding molecules, including folate-independent folate receptor binding ligands, other vitamins, peptide ligands identified from library screens, tumor-specific peptides, tumor-specific Specifically expressed in enemy aptamers, tumor-specific carbohydrates, tumor-specific monoclonal or polyclonal antibodies, Fab or scFv (ie, single chain variable region) fragments of antibodies (eg, EphA2, or metastatic cancer cells) Fab fragments of antibodies directed to or only used other proteins, small organic molecules derived from combinatorial libraries, homologous polypeptides to growth factors (eg, EGF, FGF, insulin, insulin-like growth factor), somatostatin and its Analogs, transferrins, lipoprotein complexes, bile salts, selectins, steroid hormones, Arg-Gly-Asp bearing peptides, retinoids, various Small organics containing galectins, δ-opioid receptor ligands, cholecystokinin A receptor ligands, ligands specific for angiotensin AT1 or AT2 receptors, peroxysomal proliferator-activated receptor γ ligands, β-lactam antibiotics, and antimicrobials Molecules, other molecules or fragments thereof that specifically bind to receptors preferentially expressed on the surface of tumor cells or infectious organisms are included. In particular, in the case of a ligand that binds an infectious organism, any molecule known in the art as preferentially binding to a microorganism, such as an antibiotic or other drug, corresponds. The invention also applies to molecules designed to fit into the binding pockets of specific receptors, for example antimicrobials, based on the crystal structure of the receptor or other cell surface protein, wherein the receptor is a tumor, bacteria, virus, mycoplasma, fungus, parasite or It is preferentially expressed on the surface of other pathogens. In addition, in preferred embodiments of the present invention, ligands that bind to any tumor antigen or other molecule that is preferentially expressed on the surface of the tumor cell can be used.
    [26] The binding site for the ligand may include a receptor for any molecule that can specifically bind to the receptor, where the receptor or other protein is preferentially expressed in a pathogenic cell population, for example opioid peptides, hormones, antibodies, It is a receptor for carbohydrates, small organic molecules, as well as growth factors, vitamins and peptides. The binding site may be a binding site for any molecule known in the art as being preferentially present in the microorganism, for example antibiotics or other drugs. For example, such binding sites are binding sites for β-lactam antibiotics such as penicillin in bacterial cell walls, or binding sites for antiviral agents uniquely present on the surface of the virus. The present invention also applies to binding sites for ligands, eg, antibacterial agents, designed to be suitable for the binding site of a specific receptor based on the crystal structure of the receptor, where the receptor is preferentially expressed on the surface of pathogenic cells or organisms. In addition, tumor-specific antigens may function as binding sites for ligands in the methods of the invention. An example of a tumor-specific antigen that can function as a binding site for a ligand-immunogen conjugate is an extracellular epitope of a protein member of the Ephrin family, such as EphA2. EphA2 expression is limited to cell-cell junctions in normal cells, but in metastatic tumor cells, EphA2 is distributed over the entire cell surface. Thus, EphA2 is a ligand-immunogen specific for metastatic cancer cells because EphA2 can be used for binding to Fab fragments of antibodies conjugated to immunogens, for example in metastatic cells, but not for binding to Fab fragments in normal cells. Resulting in the conjugate. The present invention also contemplates the use of ligand-immunogen conjugates to maximize the targeting of pathogenic cells in elimination by the innate immune response.
    [27] Suitable immunogens for use in the present invention are immunogens that can activate receptor members of the toll-like lineage (eg, TLR2, TLR4, TLR5, TLR6, TLR9, TLR10) and induce an innate immune response. Suitable immunogens for use in the present invention include structures essential for binding to ligands capable of activating toll-like receptors, for example, muramyl dipeptides, lipopeptides, lipoproteins, lipoarabinomannans and toll-like receptors. Characterized by lipid A, LPS, peptidoglycan, Zymosan (yeast cell wall preparation), GPI-fixed protein, soluble tuberculosis factor, Taxol, F protein of respiratory syncytial virus, flagellin, nucleotides (e.g. , CpG nucleotides), lipoteichoic acid, N-formyl Met, mannan, mannoprotein, and the like. In addition, ligands endogenously present in the host animal, such as fibronectin, fibrinogen, other extracellular matrix components, factors released as a result of cell death or damage, thermal shock heat shock proteins (eg, hsp 60) may also be used. In one embodiment, the ligand-immunogen conjugate is a conjugate in which the ligand is a vitamin and the immunogen can activate receptor members of the toll-like lineage.
    [28] Ligands and immunogens of the invention can be conjugated by any method known in the art to form complexes. This may include covalent, ionic or hydrogen bonding of the ligand and immunogen either directly or indirectly through a linking group such as a divalent linker. Typically, conjugates are formed by covalent bonds of ligands and immunogens through the formation of acid, aldehyde, hydroxy, amino or hydrazeoamide amides, esters or imino bonds in the individual components of the complex. In a preferred embodiment, the ligand is folic acid, a folic acid analog or any other folate-receptor binding molecule, wherein the folate ligand uses trifluoroacetic anhydride to prepare the γ-ester of folic acid through a proteyl azide intermediate. Conjugate to the immunogen. This preferred process results in the synthesis of folate ligands conjugated to the immunogen only through the γ-carboxyl groups of the glutamic acid functional groups of the folate, where the γ-conjugate binds to the folate receptor with high affinity to the α-conjugates and γ-conjugates. Avoid the formation of mixtures. Alternatively, pure α-conjugates can be made from intermediates, where the γ-carboxyl groups are selectively blocked and the α-conjugates are formed, after which the γ-carboxyl groups are decomposed by organic synthesis protocols and procedures known in the art. Is blocked. Other vitamins may also be used as ligands for making the conjugates according to the invention. For example, ligand-immunogen conjugates can be formed with vitamins such as biotin, riboflavin, thiamine, vitamin B 12 as well as folate (US Patent. 5,108,921, 5,416,016, 5,635,382). Alternatively, folate receptor binding ligands independent of folate may be used.
    [29] Ligand-immunogen conjugates of the present invention enhance endogenous immune response-mediated clearance of pathogenic cell populations. Endogenous immune responses include cell-mediated immune responses and other immune responses inherent in host animals. For example, endogenous immune responses may play an important role in innate immune responses by functioning as antigen-presenting cells and involve immune cells, such as macrophages and dendritic cells, that express toll-like receptors. The transferred ligand-immunogen conjugate serves as a cross-linking agent for recruiting immune cells expressing toll-like receptors into pathogenic cells. Direct recognition of these metastasized immunogens by toll-like receptors expressed on the surface of antigen-presenting cells results in the differentiation of cytokines such as IL-12 and IL-18 that induce differentiation of naïve T cells into T H 1 cells. Can result in production. Alternatively, immunogens may indirectly activate toll-like receptors through interactions with auxiliary proteins and ultimately induce an innate immune response involving differentiation of naïve T cells into T H 1 cells. Activation of toll-like receptors provides an innate mechanism by which ligand-immunogen conjugates preferentially activate cell-mediated immunity against pathogenic cell populations.
    [30] Immunogens result in presentation to naïve T cells either by antigen-presenting cells via interaction with toll-like receptors, or as MHC or MHC-like antigens (eg, CD1) at the surface of the immunogen's processing and antigen-presenting cells. It may be internalized by phagocytosis or endocytosis. As a result of antigen presentation, antigen-presenting cells induce differentiation of T cells into T H 1 cells, but T H 2 responses may also be induced. Polarization of the immune response into the T H 1 or T H 2 response depends on cytokines released during the interaction of T cells with antigen-presenting cells. The T H 1 and T H 2 responses are co-ordinated with the release of promoting cytokines (eg, IL-12, which induces a T H 1 response, and IL-4, which induces an IL-18 and T H 2 response) from antigen-presenting cells. This is the result of antigen presentation. In addition, agonist cytokines released from T H 1 cells (eg IFN-γ) and T H 2 cells (eg IL-4, IL-5, IL-10, IL-13) may further promote T cells. Can be. In addition, innate immune responses may utilize cytokine secretion that regulates processes such as replication and migration of immune cells.
    [31] Toll-like receptor ligands can induce humoral responses as a result of innate immune induction. In such cases, toll-like receptor ligands may bind to toll-like receptors in dendritic cells (type antigen-presenting cells) to promote maturation of dendritic cells. To induce adaptive immunity (eg, humoral response) by stimulating T lymphocytes to secrete cytokines that promote humoral responses.
    [32] When adaptive immunity (ie, a humoral response) is induced as a result of innate immunity, antibodies are directed to tumor cells or infectious organisms by defecting ligand-immunogen conjugates that preferentially bind to invading cells or organisms, and pathogenic cells complement It is thought to be killed by mediated lysis, ADCC, antibody-dependent phagocytosis, or antibody clustering of receptors. This cytotoxic process involves cells of other types of immune response, such as cell-mediated immunity and attracted antigen-presenting cells, phagocytizing unwanted cells and presenting the native tumor antigens of foreign pathogens to the immune system to retain these antigens. Or it may involve a secondary reaction to remove the organism.
    [33] In order to enhance endogenous immune response-mediated clearance of the pathogenic cell population, at least one additional composition containing a therapeutic factor may be administered in combination with, or as an adjunct to, the methods described above, or one or more other therapeutic factors. Can be. The therapeutic factor is a compound that can promote an endogenous immune response, or another therapeutic factor that can compensate for the effect of the administered ligand-immunogen complex. The method of the present invention, in addition to the conjugates described above, cytokines or immune cell growth factors such as interleukin 1-18, stem cell factor, basic FGF, EGF, G-CSF, GM-CSF, FLK-2 ligand, HILDA, Administering to a host a compound or composition capable of promoting an endogenous immune response including but not limited to MIP-1α, TGFα, TGFβ, M-CSF, IFNα, IFNβ, IFNγ, soluble CD23, LIF, and combinations thereof It can be carried out.
    [34] A therapeutically effective combination of these cytokines can also be used. In a preferred embodiment, about 5000 IU / dose / day to 500,000 IU / dose / day of IL-2 as a multiple dose daily regimen and about 7500 IU / dose / day to 150,000 IU as an example multiple dose daily regimen IFN-α in / dose / day is used in combination with folate-muramil dipeptides to remove these cells from host animals, including pathogenic cell populations. In another preferred embodiment, the therapeutically effective amount of IL-12 and IFN-α is combined with a ligand-immunogen conjugate, and in another preferred embodiment, the therapeutically effective amount of IL-15 and IFN-α is combined with a ligand-immunogen conjugate. Use together. In another preferred embodiment, IL-2, IFN-α or IFN-γ, GM-CSF are combined with ligand-immunogen conjugates. Suitably, the therapeutic factors used, such as IL-2, IL-12, IL-15, IL-18, IFN-α, IFN-γ, GM-CSF, combinations thereof, are natural killer cells and / or Activate T cells (T H 1 cells). Alternatively, the therapeutic factor or combinations thereof, for example, an interleukin in combination with interferons and GM-CSF is to enable such other immune fruition body cells, such as macrophages, B cells, neutrophils, LAK cells or, T H 2 cells can be activated (eg, IL-4, IL-10, IL-11). The present invention also contemplates the use of other effective combinations of cytokines, including combinations of other interleukins, interferons, colony stimulating factors.
    [35] In the method of the present invention, a chemotherapeutic agent which is itself cytotoxic and enhances tumor permeability, may be used in combination with a ligand-immunogen conjugate, and such chemotherapeutic agents include adrenocorticoids, alkylating agents, antiandrogens, antiestrogens, androgens, and estrogens. Already approved chemotherapeutic agents such as; Anti-metabolites such as cytosine arabinoside, purine analogs, pyrimidine analogs, methotrexate; Busulfan; Carboplatin; Chlorambucil; Cisplatin and other platinum compounds; Tamoxifen; Taxol; Cyclophosphamide; Plant alkaloids; Prednisone; Hydroxyurea; Teniposide; Antibiotics such as mitomycin C and bleomycin; Nitrogen mustard; Nitrosourea; Vincristine; Vinblastine; Inflammatory and proinflammatory agents. Other therapeutic agents that may be adjuvant to the conjugates according to the invention include penicillin, cephalosporins, vancomycin, erythromycin, syndamycin, rifampin, chloramphenicol, aminoglycosides, gentamicin, amphotericin B, acyclovir, triple Uridine, gancyclovir, zidobudine, amantadine, ribavirin, any other known antimicrobial compound, and the like.
    [36] Removal of the pathogenic cell population consists of reducing or eliminating tumor nodules of the pathogenic organism or pathogenic cells resulting in a therapeutic response. In the case of tumors, removal is the removal of primary tumor cells or cells that have metastasized or are in the process of being separated from the primary tumor. Prophylactic treatment that inhibits tumor recurrence following tumor removal by any treatment mode, including surgical removal of the tumor, radiotherapy, chemotherapy, biotherapy, is also intended. Prophylactic treatment is initial treatment with a ligand-immunogen conjugate, eg, a multi-dose daily regimen, or additional treatment or a series of treatments days or months after the initial treatment.
    [37] The present invention also relates to pharmaceutical compositions containing ligand-immunogen conjugates that "label" a pathogenic cell population in a host animal for specific removal by an endogenous immune response. Optionally, the composition further contains a compound capable of promoting an endogenous immune response, wherein the compound does not bind to the ligand-immunogen conjugate. The compound is effective for enhancing the removal of pathogenic cells. The pharmaceutical composition contains a therapeutically effective amount of a ligand-immunogen conjugate and a therapeutic factor, which therapeutic agent is a cytokine such as IL-2, IL-4, IL-10, IL-11, IL-12, IL -15, IL-18; Combinations of cytokines such as IL-2, IL-4, IL-10, IL-11, IL-12, IL-15, IL-18 and interferon such as IFN-α, IFN-γ; Interferon, interleukin, colony stimulating factor, for example a combination of GM-CSF.
    [38] The unit daily dose of ligand-immunogen conjugates is highly dependent on the combination of other therapies such as host condition, disease state to be treated, molecular weight of conjugate, route of administration, tissue distribution, radiotherapy. The effective amount administered to the patient is based on body surface area, patient weight, and physician's assessment of patient condition. The effective amount is approximately 1 ng / kg to 1 mg / kg, more preferably approximately 1 μg / kg to 500 µg / kg, most preferably approximately 1 µg / kg to 100 µg / kg.
    [39] An effective regimen for administering ligand-immunogen conjugates and therapeutic factors can be used. For example, ligand-immunogen conjugates and therapeutic factors can be administered in a single dose or in a multi-dose daily regimen. In addition, differential agents, such as 1 to 3 day regimens per week, may be used as an alternative to daily treatment, where such intermittent or differential regimen is considered equivalent to daily treatment and is within the scope of the present invention. In a preferred embodiment of the invention, the host is treated with multiple injections of ligand-immunogen conjugates and therapeutic factors to eliminate pathogenic cell populations. In one embodiment, the host is treated by several (preferably 2-50) injections of ligand-immunogen conjugates, eg, at 12-72 hour intervals or at 48-72 hour intervals. Additional injections of ligand-immunogen conjugates can be administered to patients days or months after the first injection, which prevents the recurrence of the disease. Alternatively, the first injection of ligand-immunogen conjugate can prevent the recurrence of the disease.
    [40] The therapeutic factor may be administered to the host animal before, after or concurrently with the ligand-immunogen conjugate and is administered as part of the same composition containing the ligand-immunogen conjugate or as part of a composition different from such conjugate. Such therapeutic compositions containing a therapeutically effective amount of a therapeutic factor can be used in the present invention. Furthermore, more than one type of ligand-immunogen conjugate can be used. For example, host animals can be treated with muramyl dipeptides and Taxol or CpG nucleotides linked to the same or different ligands in a combination protocol. In the case of chemotherapeutic and antimicrobial agents, the therapeutic factors may be administered in suboptimal doses with ligand-immunogen conjugates in combination therapy to avoid the development of resistance to chemotherapeutic or antimicrobial agents in the host animal.
    [41] Suitably, the ligand-immunogen conjugate and the therapeutic factor are administered by extra-intestinal injection, such as intraperitoneal injection, subcutaneous injection, intramuscular injection, intravenous injection or subarachnoid injection. Ligand-immunogen conjugates and therapeutic factors can also be delivered to a patient using a slow pump. Examples of extra-intestinal dosage forms are aqueous solutions of the active ingredient dissolved in isotonic saline, 5% glucose, or other known pharmaceutically acceptable liquid carriers such as alcohols, glycols, esters, amides. The extra-intestinal composition according to the present invention may be in the form of a reconstitutable lyophilizate containing a dose of ligand-immunogen conjugate and a therapeutic factor. In a preferred aspect of the present invention, ligand-immunogen conjugates are prepared in a variety of long-term release forms known in the art, for example U.S. Patent No. 4,713,249; 5,266,333; It may be administered in a biodegradable carbohydrate matrix disclosed in 5,417,982.
    [42] Oral ingestion may also be used to administer ligand-immunogen conjugates and therapeutic factors, such as syrups, sprays, other liquid dosage forms, gel-seals, or capsules or caplets. Buccal and sublingual administration is a pharmaceutically acceptable liquid dosage form, such as a syrup or spray, or a saliva-soluble dosage form that forms a saliva solution in contact with the oral and pharyngeal mucosa in the patient's mouth. Contacting the patient's mouth and pharyngeal mucosa. Typical saliva-soluble forms are lozenge tablets, tablets and the like.
    [43] In the present invention, the ligand-immunogen conjugate and the therapeutic factor to be administered buccally or sublingually are administered to the patient in a dosage form suitable for facilitating contact of the patient's mouth with the pharyngeal mucosa. Thus, the dosage form may be in the form of a liquid solution, such as syrup, spray or other liquid dosage form, administered and used in a patient in a manner that facilitates contact with oral mucosal tissue. Alternatively, the conjugate may be administered by oral ingestion with a therapeutic factor, wherein the compound is prepared in a syrup that is swallowed by the patient and does not remain in the mouth. Syrups for this purpose are prepared with additional calorie sweeteners or non-calorie sweeteners, flavor oils, pharmaceutically acceptable surfactants / dispersants, with or without aroma, as a base in a buffered aqueous solution. can do. Other liquid dosage forms, including solutions or sprays, can be prepared in a similar manner and can be administered by buccal, sublingual or oral ingestion.
    [44] Suitably the conjugates and therapeutic factors administered buccally / sublingually in the present invention are prepared in solid dosage forms, for example lozenge tablets or tablets. Such formulations contain a saliva-soluble carrier and may optionally contain the desired excipients such as buffers or tableting aids.
    [45] The lozenge tablets used in the present invention can be prepared by techniques known in the art, for example, to form compressed tablets, wherein the conjugates and the therapeutic factors are dispersed in a compressible solid carrier and a lubricant (eg magnesium-stearate) It is optionally mixed with a suitable tableting aid such as) and compressed into tablets. Solid carrier components suitable for such tableting formulations are saliva-soluble solids, such as cold water-soluble starch, monosaccharides or disaccharides, which are readily soluble in the mouth. The pH of the aforementioned formulations is between 4 and 8.5. Lozenge tablets used in the present invention may also be prepared using other unit dosage form techniques known in the art.
    [46] Tablets used in the present invention can be prepared in a manner similar to that found in the manufacture of lozenge tablets, or by other techniques known in the art to form compressed tablets such as chewable vitamins. Solid carrier components suitable for tableting are mannitol, microcrystalline cellulose, carboxymethyl cellulose, dibasic calcium phosphate and the like.
    [47] Solid dosage forms for oral ingestion administration include dosage forms such as caplets, capsules and gel-seals. Such solid dosage forms can be prepared using standard tableting protocols and excipients that provide conjugates and therapeutic factor-containing capsules, caplets or gel-seals. Solid dosage forms used in the present invention, including lozenges and tablets, may be in a form suitable for sustained release.
    [48] Example 1 Effect of Folate-Muramil Dipeptide Conjugates on Survival of Lung Cancer Transplanted Mice
    [49] Female Balb / c mice were injected on day 0 with 5 × 10 5 M109 cells, a syngeneic lung cancer cell line expressing high levels of folate receptors. To reduce the time needed to obtain long-term survival data, these tumor cells were implanted intraperitoneally close to the liver. The cancer site then settled and grew. These animals were then injected with PBS (control) at 7, 8, 9, 11 and 14 days after tumor cell transplantation or with folate-muramil dipeptide (MDP) (15 nmoles / kg), IL- 2 (5,000 IU / dose), IFN-α (25,000 U / dose) were co-injected. Folate was conjugated to MDP via gamma carboxyl-linked ethylenediamine crosslinking. Other animals were injected with 80 nmoles / kg MDP, 15 nmoles / kg folate-MDP, or 80 nmoles / kg MDP, IL-2, IFN-α at the concentrations specified above. The efficacy of this immunotherapy was assessed by monitoring survival as a function of time in folate-MDP treated mice compared to control animals. As shown in FIG. 1, control mice all died on day 21, whereas mice treated with MDP-folate + IL-2 + IFN-α survived up to 40 days. In addition, folate-MDP in combination with IL-2 and IFN-α shows a strong synergy in the ability to promote long-term survival of tumor-bearing mice, while the combination of IL-2 and IFN-α results in long-term survival of mice. Had little effect on folate and folate-MDP alone had little effect on the survival of mice.
    [50] Example 2: Effect of Folate-CpG Conjugates on Survival of Lung Cancer Transplanted Mice
    [51] Female Balb / c mice were injected on day 0 with 5 × 10 5 M109 cells, a syngeneic lung cancer cell line expressing high levels of folate receptors. To reduce the time needed to obtain long-term survival data, these tumor cells were implanted intraperitoneally close to the liver. The cancer site then settled and grew. These animals were then injected with PBS (control), CpG (1500 nmoles / kg), or folate-CpG (1500 nmoles / kg) on days 7 and 8 after tumor cell transplantation. Folate was conjugated to 5'-amino CpG via peptide crosslinking. The efficacy of this immunotherapy was assessed by monitoring survival as a function of time. As shown in FIG. 2, the control mice all died up to 25 days, whereas mice treated with CpG survived up to 44 days and mice treated with folate-CpG survived up to 50 days.
    权利要求:
    Claims (38)
    [1" claim-type="Currently amended] A method of enhancing endogenous immune response-mediated specific clearance of a pathogenic cell population in a host animal comprising a pathogenic cell population, wherein the member of the cell population has available binding sites for ligands, Method comprising the steps of:
    A ligand-immunogen conjugate composition containing a complex of ligand and immunogen is administered to the host, wherein the immunogen can activate toll-like receptors, wherein the ligand is a vitamin or derivative or analog thereof;
    A therapeutic factor is administered to the host, where the therapeutic factor is a compound capable of promoting an endogenous immune response, and the compound does not bind to the ligand-immunogen conjugate.
    [2" claim-type="Currently amended] The method of claim 1, wherein the pathogenic cell population is a cancer cell population.
    [3" claim-type="Currently amended] The method of claim 2, wherein the cancer cell population forms a tumor.
    [4" claim-type="Currently amended] The method of claim 1, wherein the pathogenic cell population is an exogenous pathogen.
    [5" claim-type="Currently amended] 5. The method of claim 4, wherein the exogenous pathogen is selected from bacteria, fungi, viruses, mycoplasmas, parasites.
    [6" claim-type="Currently amended] The method of claim 1, wherein the pathogenic cell population is an endogenous cell population comprising exogenous pathogens.
    [7" claim-type="Currently amended] The method of claim 1 wherein the ligand component of the ligand-immunogen conjugate is a vitamin capable of specifically binding to cell membrane receptors.
    [8" claim-type="Currently amended] 8. The method of claim 7, wherein the ligand component of the ligand-immunogen conjugate is selected from folic acid and other folate receptor-binding ligands.
    [9" claim-type="Currently amended] The method of claim 1, wherein the ligand component of the ligand-immunogen conjugate is selected from folic acid and other folate receptor-binding ligands.
    [10" claim-type="Currently amended] The method of claim 1, wherein the ligand is chemically complexed to the immunogen via covalent bonds, ionic bonds, or hydrogen bonds.
    [11" claim-type="Currently amended] The method of claim 10, wherein the ligand is a folic acid analog having a glutamyl moiety covalently linked to the immunogen only via the glutamyl γ-carboxyl moiety.
    [12" claim-type="Currently amended] The method of claim 10, wherein the ligand is a folic acid analog having a glutamyl moiety covalently linked to the immunogen only via the glutamyl α-carboxyl moiety.
    [13" claim-type="Currently amended] 12. The method of claim 11, wherein the covalent bond between the immunogen and the ligand is a direct covalent bond to the immunogen, or a covalent bond via a divalent linker.
    [14" claim-type="Currently amended] The method of claim 12, wherein the covalent bond between the immunogen and the ligand is a direct covalent bond to the immunogen, or a covalent bond via a bivalent linker.
    [15" claim-type="Currently amended] 2. The ligand component of claim 1, wherein the ligand component of the ligand-immunogen conjugate is a small organic molecule capable of binding to a receptor, said receptor being preferentially expressed, uniquely expressed or overexpressed on the surface of a pathogenic cell population. How to.
    [16" claim-type="Currently amended] The method of claim 15, wherein the small organic molecule is an antimicrobial agent.
    [17" claim-type="Currently amended] The method of claim 16, wherein the antimicrobial agent is β-lactam antibiotics.
    [18" claim-type="Currently amended] The method of claim 1, wherein the binding site for the ligand component of the ligand-immunogen conjugate is an antigen that is preferentially expressed, uniquely expressed or overexpressed in metastatic cancer cells.
    [19" claim-type="Currently amended] The method of claim 18, wherein the ligand binding site is EphA2.
    [20" claim-type="Currently amended] The method of claim 1, wherein the immunogen is muramyl dipeptide.
    [21" claim-type="Currently amended] The method of claim 1, wherein the immunogen is a nucleotide.
    [22" claim-type="Currently amended] The method of claim 21, wherein the nucleotide is CpG.
    [23" claim-type="Currently amended] The method of claim 1, wherein the therapeutic factor consists of cytokines.
    [24" claim-type="Currently amended] The method of claim 23, wherein the therapeutic factor consists of IL-2, IL-4, IL-10, IL-11, IL-12, IL-15, IL-18, or a combination thereof.
    [25" claim-type="Currently amended] The method of claim 23, wherein the therapeutic factor is IL-2, IL-4, IL-10, IL-11, IL-12, IL-15, IL-18, or a combination thereof in combination with IFN-α or IFN-γ. And configured.
    [26" claim-type="Currently amended] The method of claim 23, wherein the therapeutic factors are IL-2, IL-4, IL-10, IL-11, IL-12, IL-15, IL-18, IFN-α, IFN-γ, GM-CSF, these It is selected from the combination of.
    [27" claim-type="Currently amended] The method of claim 1, wherein the endogenous immune response is a cell-mediated immune response.
    [28" claim-type="Currently amended] In a method of enhancing endogenous immune response-mediated specific clearance of a pathogenic cell population in a host animal comprising a pathogenic cell population, said cell population preferentially expresses, uniquely expresses or overexpresses a vitamin receptor, The method comprises the following steps:
    A composition containing a ligand covalently linked to an immunogen is administered to the host, wherein the immunogen can activate toll-like receptors, and the ligand is a vitamin or derivative or analog thereof.
    [29" claim-type="Currently amended] Pharmaceutical composition consisting of:
    Therapeutically effective amount of ligand-immunogen conjugate, wherein the immunogen can activate toll-like receptors and the ligand is a vitamin or derivative or analog thereof;
    Therapeutic factor, wherein the therapeutic factor is a compound capable of promoting an endogenous immune response, and the compound does not bind to ligand-immunogen conjugates;
    Pharmaceutically acceptable carrier.
    [30" claim-type="Currently amended] 30. The pharmaceutical composition according to claim 29, which is in an extra enteral long release form.
    [31" claim-type="Currently amended] The pharmaceutical composition according to claim 29, which is in oral dosage form.
    [32" claim-type="Currently amended] The pharmaceutical composition of claim 29, wherein the therapeutic factor is an immunostimulator.
    [33" claim-type="Currently amended] The method of claim 32, wherein the immunostimulator is IL-2, IL-4, IL-10, IL-11, IL-12, IL-15, IL-18, IFN-α, IFN-γ, GM-CSF, A pharmaceutical composition, characterized in that the compound selected from a combination thereof.
    [34" claim-type="Currently amended] 30. The pharmaceutical composition of claim 29, wherein the ligand is folic acid or folic acid analog.
    [35" claim-type="Currently amended] The pharmaceutical composition of claim 29, wherein the immunogen is a nucleotide.
    [36" claim-type="Currently amended] 36. The pharmaceutical composition of claim 35, wherein the nucleotide is CpG.
    [37" claim-type="Currently amended] A method of treating a disease state in a host animal, wherein the disease state is mediated by activated macrophages, the macrophages retain available binding sites for ligands, and the method comprises the following steps: How to feature:
    A ligand-immunogen conjugate composition containing a complex of ligand and immunogen is administered to the host, wherein the immunogen can activate toll-like receptors.
    [38" claim-type="Currently amended] Pharmaceutical composition consisting of:
    Therapeutically effective amount of ligand-immunogen conjugate, wherein the immunogen can activate toll-like receptors and the ligand is a vitamin or derivative or analog thereof;
    Pharmaceutically acceptable carrier.
    类似技术:
    公开号 | 公开日 | 专利标题
    US20170246314A1|2017-08-31|Folate Receptor Binding Conjugates of Antifolates
    Martins et al.2015|Vaccine adjuvant uses of poly-IC and derivatives
    US10258700B2|2019-04-16|Methods and compositions for delivering active agents with enhanced pharmacological properties
    ES2741308T3|2020-02-10|T-cell switches with chimeric antigen receptors and uses thereof
    JP2018030837A|2018-03-01|Synthetic nano carrier that generates humoral and cytotoxic t lymphocyte | immune response
    JP6407208B2|2018-10-17|Synthetic nanocarrier combination vaccine
    US20180296663A1|2018-10-18|Vaccine composition
    JP6453855B2|2019-01-16|Compositions and methods for activating "interferon gene stimulator" dependent signaling
    CN104507538B|2018-04-06|The composition and method of immunotherapy for cancer
    US9468687B2|2016-10-18|Immuno gene therapy for treatment of cancer and hyperproliferative diseases
    US20190105399A1|2019-04-11|Vitamin-Receptor Binding Drug Delivery Conjugates
    US8790655B2|2014-07-29|Conjugates of synthetic TLR agonists and uses therefor
    JP6148062B2|2017-06-14|Hydrophobic core carrier composition for delivery of therapeutic agents and methods of making and using the same
    JP2017149743A|2017-08-31|Vaccination in elderly patient
    Hanson et al.2015|Nanoparticulate STING agonists are potent lymph node–targeted vaccine adjuvants
    EP2680880B1|2017-04-26|Vaccination in newborns and infants
    JP5539460B2|2014-07-02|Compositions and methods for cancer immunotherapy
    Hanagata2017|CpG oligodeoxynucleotide nanomedicines for the prophylaxis or treatment of cancers, infectious diseases, and allergies
    RU2708246C2|2019-12-05|Drug combinations
    JP6426706B2|2018-11-21|GLA monotherapy for use in cancer treatment
    US20170327537A1|2017-11-16|Binding ligand linked drug delivery conjugates of tubulysins
    Petrovsky et al.2004|Vaccine adjuvants: current state and future trends
    KR101866893B1|2018-06-14|Therapeutic use of a tlr agonist and combination therapy
    Lahiri et al.2008|Engagement of TLR signaling as adjuvant: towards smarter vaccine and beyond
    Fontana et al.2017|Delivery of therapeutics with nanoparticles: what's new in cancer immunotherapy?
    同族专利:
    公开号 | 公开日
    引用文献:
    公开号 | 申请日 | 公开日 | 申请人 | 专利标题
    法律状态:
    2001-09-28|Priority to US32579301P
    2001-09-28|Priority to US60/325,793
    2001-10-01|Priority to US32632201P
    2001-10-01|Priority to US60/326,322
    2002-06-26|Priority to US60/391,654
    2002-06-26|Priority to US39165402P
    2002-09-26|Application filed by 펄듀 리서치 파운데이션
    2002-09-26|Priority to PCT/US2002/030546
    2004-06-23|Publication of KR20040053136A
    优先权:
    申请号 | 申请日 | 专利标题
    US32579301P| true| 2001-09-28|2001-09-28|
    US60/325,793|2001-09-28|
    US32632201P| true| 2001-10-01|2001-10-01|
    US60/326,322|2001-10-01|
    US39165402P| true| 2002-06-26|2002-06-26|
    US60/391,654|2002-06-26|
    PCT/US2002/030546|WO2003028634A2|2001-09-28|2002-09-26|Method of treatment using ligand-immunogen conjugates|
    [返回顶部]