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WO2018159808A1 - Conjugué d'anticorps-médicament de l'anticorps anti-il-7r, et composition pharmaceutique de traitement du cancer ou de l'inflammation et contenant un conjugué d'anticorps-médicament de l'anticorps anti-il-7r et d'un médicament cytotoxique - Google Patents

Conjugué d'anticorps-médicament de l'anticorps anti-il-7r, et composition pharmaceutique de traitement du cancer ou de l'inflammation et contenant un conjugué d'anticorps-médicament de l'anticorps anti-il-7r et d'un médicament cytotoxique Download PDF

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WO2018159808A1
WO2018159808A1 PCT/JP2018/007985 JP2018007985W WO2018159808A1 WO 2018159808 A1 WO2018159808 A1 WO 2018159808A1 JP 2018007985 W JP2018007985 W JP 2018007985W WO 2018159808 A1 WO2018159808 A1 WO 2018159808A1
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antibody
cancer
pharmaceutical composition
steroid
cells
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Japanese (ja)
Inventor
正浩 安永
保広 松村
眞鍋　史乃
厚至 辻
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National Cancer Center Japan
RIN Institute Inc
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National Cancer Center Japan
RIN Institute Inc
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Priority to JP2019503147A priority Critical patent/JPWO2018159808A1/ja
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/47—Quinolines; Isoquinolines
    • A61K31/4738—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems
    • A61K31/4745—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems condensed with ring systems having nitrogen as a ring hetero atom, e.g. phenantrolines
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    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/47—Quinolines; Isoquinolines
    • A61K31/475—Quinolines; Isoquinolines having an indole ring, e.g. yohimbine, reserpine, strychnine, vinblastine
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    • A61K31/57—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids substituted in position 17 beta by a chain of two carbon atoms, e.g. pregnane or progesterone
    • A61K31/573—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids substituted in position 17 beta by a chain of two carbon atoms, e.g. pregnane or progesterone substituted in position 21, e.g. cortisone, dexamethasone, prednisone or aldosterone
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    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
    • A61K31/7034—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
    • A61K31/704—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin
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    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/42—Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein
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    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
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    • A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
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    • A—HUMAN NECESSITIES
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    • A61P35/00—Antineoplastic agents
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A—HUMAN NECESSITIES
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    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/24—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against cytokines, lymphokines or interferons

Definitions

  • the present invention relates to an antibody drug conjugate of an anti-IL-7R antibody (anti-IL-7 receptor antibody) and an antibody of an anti-IL-7R antibody and a cytotoxic agent for use in treating cancer or inflammation. It relates to a pharmaceutical composition comprising a drug conjugate.
  • SN-38 a topoisomerase 1 inhibitor
  • CPT-11 was developed as a compound that improves the blood translocation of SK-38.
  • CPT-11 is a prodrug of SN-38, and the conversion efficiency to the active form is high in cancer tissues of solid tumors, but is low in blood and low in blood. Therefore, CPT-11 could not be used to treat leukemia.
  • IL-7 is known as an essential cytokine for T cell generation and homeostasis (Non-patent Document 1).
  • IL-7R ⁇ moves into the cell slowly and partly returns to the cell surface, but partly decomposes (Non-patent Document 1).
  • the half-life of IL-7R ⁇ is shortened from about 24 hours to 3 hours by stimulation with IL-7 (Non-patent Document 1).
  • IL-7R is responsible for cancer steroid resistance (Non-patent Document 2).
  • Non-Patent Document 2 suggests that suppressing downstream signals of IL-7R is effective in treating steroid resistance.
  • the present invention relates to a pharmaceutical composition
  • a pharmaceutical composition comprising an antibody drug conjugate of an anti-IL-7R antibody and an antibody drug conjugate of an anti-IL-7R antibody and a cytotoxic agent for use in treating cancer or inflammation.
  • IL-7R is associated with steroid resistance and that IL-7R knockdown suppresses cell proliferation of cancer cell lines.
  • IL-7R knockdown dramatically improves the steroid sensitivity of cancer cells (especially lymphomas and leukemias), thereby increasing the cell killing effect of steroids, and steroid resistance
  • the expression of IL-7R is improved by obtaining.
  • the inventors further found that activation of NF ⁇ B occurs in steroid resistance without STAT or BCL2 activation of downstream signals of IL-7R.
  • the inventors further have improved expression of IL-7R after chemotherapy treatment with steroidal anti-inflammatory drugs, and IL-7R in cell lines that have been treated with steroidal anti-inflammatory drugs to gain steroid resistance.
  • anti-IL-7R antibodies accumulate in lysosomes when taken up by cells, and accumulate in tumor tissues and lymph nodes one week after administration.
  • the effect of the antibody blocking the binding between IL-7R and IL-7 is weak, and on the other hand, the ADC of the anti-IL-7R antibody shows a remarkable effect, And found to suppress lymphadenopathy.
  • the effect of ADC on tumors was also shown with a radioisotope-labeled anti-IL-7R antibody.
  • the present inventors have found that the symptoms of inflammatory diseases (for example, rheumatoid arthritis and ulcerative colitis) are reduced through a decrease in IL-7R positive cells of the immune system. The present invention is based on these findings.
  • a pharmaceutical composition comprising an antibody drug conjugate of an anti-IL-7R antibody or an antigen-binding fragment thereof and a cytotoxic agent, for use in treating cancer or inflammation.
  • the pharmaceutical composition according to the above (1) for use in treating inflammation.
  • the pharmaceutical composition according to (1) or (2) above wherein the inflammation is steroid-treated or steroid-resistant inflammation.
  • the pharmaceutical composition according to (2) or (3) above, wherein the inflammation is ulcerative colitis or rheumatoid arthritis.
  • a pharmaceutical composition comprising an antibody drug conjugate of an anti-IL-7R antibody or an antigen-binding fragment thereof and a cytotoxic agent, which is used for treating a disease or condition after steroid treatment or steroid resistance
  • Pharmaceutical composition for (11) A pharmaceutical composition comprising an antibody drug conjugate of an anti-IL-7R antibody or antigen-binding fragment thereof and a cytotoxic agent, which is used in combination with a steroid.
  • FIG. 1A shows the results of examining the expression level of IL-7R in various cancer cell lines by flow cytometry.
  • FIG. 1B is a graph showing that IL-7R expression level was actually decreased in various cancer cell lines by IL-7R knockdown assay.
  • FIG. 1C is a graph showing the influence on the growth of a cancer cell line in which IL-7R is knocked down.
  • FIG. 1D is a graph showing the influence of subcutaneously transplanted IL-7R on the growth of a cancer cell line knocked down in a nude mouse subcutaneous transplantation model.
  • FIG. 1E is a photomicrograph showing the viability of a cancer cell line in which IL-7R has been knocked down.
  • FIG. 1A shows the results of examining the expression level of IL-7R in various cancer cell lines by flow cytometry.
  • FIG. 1B is a graph showing that IL-7R expression level was actually decreased in various cancer cell lines by IL-7R knockdown assay.
  • FIG. 2A is a graph showing that IC50 of various cancer cell lines against dexamethasone (a kind of steroidal anti-inflammatory drug) is decreased by IL-7R knockdown.
  • FIG. 2B shows the effect of IL-7R knockdown on the resistance to dexamethasone in vitro by obtaining a dexamethasone resistant strain.
  • FIG. 2C shows that the expression of IL-7R is improved in the dexamethasone resistant strain.
  • FIG. 3 shows the results of staining various cancer cell lines and their IL-7R knockdown lines with anti-phosphorylated p65 / NF ⁇ B antibodies.
  • FIG. 4A is a diagram showing changes in gene expression before and after chemotherapy including steroids in B-cell acute leukemia.
  • FIG. 4A is a diagram showing changes in gene expression before and after chemotherapy including steroids in B-cell acute leukemia.
  • FIG. 4B is a diagram showing changes in gene expression before and after acquisition of steroid resistance by steroid treatment in B-cell acute leukemia.
  • FIG. 4C shows changes in gene expression before and after chemotherapy including steroids in T-cell acute leukemia.
  • FIG. 5A shows the results of confirming co-localization with lysosome by contacting Alexa647-labeled anti-IL-7R antibody with a cancer cell line in vitro.
  • FIG. 5B shows the results of administering a fluorescently labeled anti-IL-7R antibody to a tumor subcutaneous transplantation model and examining the accumulation of anti-IL-7R antibody in various tissues one week later.
  • FIG. 5C shows that an anti-IL-7R antibody having the ability to neutralize binding between IL-7R and IL-7 is effective against RAG2 ⁇ / ⁇ cells that survive in an IL-7-dependent manner, It shows that it has little effect on the cancer cell line CYG82.
  • FIG. 5D shows that an antibody drug conjugate (ADC) of an anti-IL-7R antibody and a cytotoxic agent exerts a killing effect on the cancer cell line CYG82.
  • FIG. 6A is a graph showing the effect of an antibody drug conjugate of an anti-IL-7R antibody and a cytotoxic agent on tumor volume increase in a subcutaneous transplantation model of a cancer cell line.
  • FIG. 6B is a graph showing the effect of an antibody drug conjugate of an anti-IL-7R antibody and a cytotoxic agent on tumor volume increase in a steroid-resistant cancer cell line subcutaneous transplantation model.
  • FIG. 7A shows the effect of IL-7R knockdown on lymphadenopathy in a subcutaneous transplantation model of cancer cell line CYG82.
  • FIG. 7B is a graph showing the effect of ADC of anti-IL-7R and SN-38 on intraperitoneal lymphadenopathy occurring through regional lymphadenopathy in a subcutaneous transplantation model of cancer cell line CYG82.
  • FIG. 8A is a graph showing the effect of a radioisotope-labeled ( 90 Y-labeled) anti-IL-7R antibody on the T-cell lymphoma cell line Sup-T1.
  • FIG. 8B shows the effect of radioisotope-labeled ( 90 Y-labeled) anti-IL-7R antibody on lung cancer cell line H2009.
  • FIG. 9 is a diagram showing the effect of anti-IL-7R on the immune system of ADC.
  • FIG. 10 shows that IL-7R expression is increased in rheumatoid arthritis and ulcerative colitis.
  • FIG. 11A shows the effect of ADC of anti-IL-7R antibody and monomethyl auristatin E (MMAE) on rheumatoid arthritis model.
  • FIG. 11B shows the effect of anti-IL-7R antibody and SN-38 ADC on rheumatoid arthritis model.
  • FIG. 11C shows the result of preparing a tissue section of a mouse forefoot lesion of a rheumatoid arthritis model and staining with hematoxylin and eosin.
  • FIG. 12A is a graph showing the effect of ADC of anti-IL-7R antibody and MMAE on ulcerative colitis model.
  • FIG. 12B is a graph showing the effect of ADC of anti-IL-7R antibody and SN-38 on ulcerative colitis model.
  • FIG. 13 shows that the colonic morphology of the ulcerative colitis model is improved with an anti-IL-7R antibody and MMAE ADC or an anti-IL-7R antibody and SN-38 ADC.
  • FIG. 14 is a graph showing the effect of ADC of anti-IL-7R antibody and MMAE or ADC of anti-IL-7R antibody and SN-38 on the histopathological image of a colon specimen removed from an ulcerative colitis model.
  • subject means a mammal, and may be a human in particular.
  • treatment is used to mean treatment (therapeutic treatment) and prevention (preventive treatment).
  • treatment means treatment, cure, prevention or amelioration of a disease or disorder or a reduction in the rate of progression of a disease or disorder.
  • prevention means reducing the likelihood of the onset of a disease or condition or delaying the onset of a disease or condition.
  • disease means a symptom for which treatment is beneficial.
  • therapeutically effective amount means the amount of a drug effective for treating (preventing or treating) a disease or condition.
  • a therapeutically effective amount of a drug reduces the rate of worsening of a symptom of a disease or condition, stops the worsening of the symptom, improves the symptom, cures the symptom, or develops or develops the symptom Can be suppressed.
  • resistant means not showing efficacy for treatment (eg, showing no significant effectiveness).
  • sensitivity means showing efficacy for treatment. Decreasing resistance has the same meaning as increasing sensitivity, and increasing resistance has the same meaning as decreasing sensitivity.
  • steroid resistance can be defined as no complete remission in the case of nephrotic syndrome, even if prednisolone is administered daily for more than 4 weeks.
  • other inflammatory / autoimmune diseases or malignant diseases such as leukemia / malignant lymphoma
  • other inflammatory / autoimmune diseases or malignant diseases such as leukemia / malignant lymphoma
  • other inflammatory / autoimmune diseases or malignant diseases such as leukemia / malignant lymphoma
  • other inflammatory / autoimmune diseases or malignant diseases such as leukemia / malignant lymphoma
  • other inflammatory / autoimmune diseases or malignant diseases such as leukemia / malignant lymphoma
  • steroids such as prednisolone, methylprednisolone
  • IL-7R means interleukin 7 receptor.
  • JAK1 and JAK3 are activated and receptor autophosphorylation occurs.
  • STAT binds to the phosphorylated receptor and is phosphorylated by JAK to dimerize. It is thought that transcriptional activation of genes is caused by dimerized STAT.
  • IL-7R has a high internalization activity of the molecule itself, and is internalized from the membrane and decomposed or recycled again to be exposed on the membrane.
  • blocking means neutralizing the binding of two proteins. Blocking includes partial dissociation in addition to complete dissociation. For example, it can be said that blocking occurs when the binding of two proteins is reduced by 50% or more.
  • steroid means steroidal anti-inflammatory drugs (SAIDs).
  • SAIDs steroidal anti-inflammatory drugs
  • glucocorticoids or their derivatives are mainly active ingredients.
  • GR ⁇ glucocorticoid receptor
  • GR ⁇ dissociates from hsp90, forms a dimer, moves into the nucleus, and regulates transcription.
  • Synthetic steroids include cortisols such as hydrocortisone and hydrocortisone succinate; prednisolones such as prednisolone, methylprednisolone and methylprednisolone succinate; triamcinolones such as triamcinolone and triamcinolone acetonide; dexamethasone and betamethasone. Both are derivatives of glucocorticoid and function in the cell by the same mechanism as glucocorticoid.
  • antibody drug conjugate means a substance in which a monoclonal antibody or an antigen-binding fragment thereof (hereinafter sometimes simply referred to as “antibody or the like”) is linked to a drug.
  • ADC a monoclonal antibody or the like and a drug can be linked via an appropriate linker.
  • the ADC binds to a membrane component on the cell membrane (for example, a transmembrane protein such as a receptor), is taken into the cell by endocytosis or internalization, is separated from the antibody or the like, and is released into the cell.
  • the linker By introducing a cleavable linker between the antibody and the drug in the cell, the linker can be cleaved in the cell, for example, in the endosome, and the drug can be released from the antibody and released into the cytoplasm. is there.
  • a cytotoxic agent is used as a drug, it is possible to kill cells to which the drug has been delivered.
  • a chemotherapeutic agent, a radioisotope, and a toxin can be used as the cytotoxic agent.
  • IL-7R expression is improved in many tumor cells.
  • tumor growth was greatly suppressed by reducing the expression of improved IL-7R.
  • IL-7R ADCs showed a strong killing ability.
  • antibodies capable of blocking the binding between IL-7R and IL-7 had little effect on cell survival. Therefore, as long as the anti-IL-7R antibody used in the ADC of the present invention binds to IL-7R on the cell membrane, the ability to block the binding between IL-7R and IL-7 is not questioned. It may be an antibody that can block the binding to -7, or an antibody that does not block the binding between IL-7R and IL-7.
  • an antibody drug conjugate of an anti-IL-7R antibody or antigen-binding fragment thereof and a cytotoxic agent is provided.
  • an antibody that recognizes IL-7R expressed on the cell membrane can be used as the anti-IL-7R antibody.
  • the antibody or antigen-binding fragment thereof and the cytotoxic agent in the antibody drug conjugate are linked via a linker.
  • cytotoxic agents include chemotherapeutic agents (for example, anticancer agents such as commercially available anticancer agents such as auristatin (auristatin E, auristatin F phenylenediamine (AFP), monomethyl auristatin E, monomethyl auristatin).
  • Statins F and their derivatives maytansinoids DM1 and DM4 and their derivatives), camptothecin (SN-38, topotecan and exotecan and their derivatives), DNA minor groove binders (enzyin, lexitropsin, duocarmycin) And their derivatives), taxanes (paclitaxel and docetaxel and their derivatives), polyketides (discodermolide and their derivatives), anthraquinones (mitoxantrone and their derivatives), benzodiazepines (pyrrolobenzodiazepine, indolinobenzo) Azepine, and oxazolidinobenzodiazepines and their derivatives), vinca alkaloids (vincristine, vinblastine, vindesine, and vinorelbine and their derivatives), doxorubicins (doxorubicin, morpholino-doxorubicin, and cyanomorpholino-doxorubicin and their derivatives), Cardiac glycosides (digit
  • the linker can be appropriately selected and synthesized by those skilled in the art in the production of ADC.
  • the linker can be a cleavable linker.
  • cleavable linkers include peptide linkers such as valine-citrulline (Val-Cit) and phenylalanine-lysine (Phe-lys) linkers, and hydrazone linkers that cleave in a pH-dependent manner.
  • Cleaveable linkers also include linkers that contain carbamate or ester bonds, which can be enzymatically degraded intracellularly. These linkers may be used in combination.
  • the bond between the antibody and the linker can be linked to, for example, a sulfhydryl group of the antibody via a maleimide group.
  • the linker may contain a polyethylene glycol block as necessary.
  • the present invention also provides a pharmaceutical composition for use in treating cancer, comprising an antibody drug conjugate of an anti-IL-7R antibody or antigen-binding fragment thereof and a cytotoxic agent.
  • Many cancer cells are positive for IL-7R. Accordingly, many cancers can be treated by the medicament of the present invention.
  • the cancer may be an IL-7R-positive cancer or a cancer that has been confirmed to be IL-7R-positive. it can.
  • IL-7R positive cancers include leukemia, lymphoma, lung cancer, pancreatic cancer, head and neck cancer, prostate cancer, bladder cancer, breast cancer, esophageal cancer, stomach cancer, colon cancer, uterine cancer, ovary Cancer, skin cancer, thyroid cancer, thymic cancer, kidney cancer, testicular cancer, penile cancer, liver cancer, biliary tract cancer, brain tumor, bone soft tissue tumor, retroperitoneal tumor, angio-lymphatic sarcoma , And these metastatic cancers (eg, metastatic solid tumors). Many cancers, such as leukemia, lymphoma, advanced cancer, and metastatic cancer, are positive for IL-7R. Whether IL-7R is positive or negative can be easily determined using techniques such as immunostaining and FACS using an anti-IL-7R antibody.
  • the pharmaceutical composition of the present invention can be used for treating IL-7R-positive cancer having resistance to a therapy that blocks the binding between IL-7R and IL-7.
  • IL-7R is involved in imparting steroid resistance.
  • IL-7R is positive in steroid resistant cells.
  • a pharmaceutical composition for use in treating a steroid resistant disease or condition comprising an antibody drug conjugate of an anti-IL-7R antibody or antigen-binding fragment thereof and a cytotoxic agent.
  • a pharmaceutical composition comprising a gate is provided.
  • a pharmaceutical composition for use in reducing steroid resistance in a disease or condition comprising an antibody drug conjugate of an anti-IL-7R antibody or antigen-binding fragment thereof and a cytotoxic agent.
  • a pharmaceutical composition is provided.
  • the steroid resistant disease or condition is cancer or inflammation. Furthermore, according to the present invention, it was also revealed that expression of IL-7R is improved after steroid treatment. Particularly after steroid treatment, IL-7R positive cells are more likely to survive, increasing the effectiveness of the conjugates of the invention. Even if IL-7R negative cells survive (for example, in patients after steroid treatment), a therapeutic effect can be expected. This is because the effect of killing IL-7R negative cells in the vicinity of IL-7R positive cells can be expected by the bystander effect of PDC.
  • a pharmaceutical composition for use in treating a disease or condition such as cancer or inflammation in a subject who has received steroid therapy wherein the anti-IL-7R antibody or antigen-binding property thereof is used.
  • Pharmaceutical compositions comprising antibody drug conjugates of fragments and cytotoxic agents are provided.
  • a pharmaceutical composition for use in treating a steroid resistant disease or condition, wherein the anti-IL-7R antibody or antigen-binding fragment thereof and cytotoxicity are used in combination with a steroid.
  • Pharmaceutical compositions comprising an antibody drug conjugate with an agent are provided.
  • a pharmaceutical composition for use in reducing steroid resistance of a disease or condition or increasing steroid sensitivity of the disease or condition
  • a pharmaceutical composition comprising an antibody drug conjugate of a sex fragment and a cytotoxic agent is provided.
  • the subject can be a subject with a steroid resistant disease or condition.
  • an antibody drug conjugate of an anti-IL-7R antibody or an antigen-binding fragment thereof and a cytotoxic agent can suppress lymphadenopathy in cancer subjects. Therefore, according to the present invention, a pharmaceutical composition for use in suppressing lymphadenopathy in a cancer subject, comprising an anti-IL-7R antibody or an antigen-binding fragment thereof and an antibody of a cytotoxic agent A pharmaceutical composition comprising a drug conjugate is provided.
  • an antibody drug conjugate of an anti-IL-7R antibody and a cytotoxic agent showed a symptom reducing effect.
  • IL-7R positive cells are strongly involved in the onset or maintenance of inflammatory diseases, and killing IL-7R positive cells in an inflammatory state is useful for alleviating inflammation. It means that there is.
  • a pharmaceutical composition for use in treating an inflammatory disease comprising an antibody drug conjugate of an anti-IL-7R antibody or antigen-binding fragment thereof and a cytotoxic agent A pharmaceutical composition is provided.
  • inflammatory diseases include collagen disease, autoimmune disease, type I diabetes, multiple sclerosis, systemic lupus erythematosus, allergic disease, rejection after organ transplantation, and graft versus host Diseases are also included. Inflammatory diseases also include inflammatory bowel diseases such as Crohn's disease.
  • autoimmune diseases and collagen diseases include Sjogren's syndrome, scleroderma, dermatomyositis, polyarteritis nodosa, mixed connective tissue disease, Corgan syndrome, polymyalgia rheumatica, adult Still's disease, giant cells Arteritis, antiphospholipid antibody syndrome, Guillain-Barre syndrome, myasthenia gravis, autoimmune hepatitis, autoimmune pancreatitis, primary biliary cholangitis, Takayasu arteritis, Goodpasture syndrome, rapidly progressive glomeruli Nephritis, autoimmune hemolytic anemia, autoimmune neutropenia, idiopathic thrombocytopenic purpura, Graves' disease, Hashimoto's disease, primary hypothyroidism, idiopathic Addison's disease, pemphigus vulgaris, pustular psoriasis Psoriasis vulgaris, pemphigoid, gestational herpe
  • lymphocytes are IL-7R positive in many inflammations. Specifically, many inflammatory diseases caused by these cells because they are positive in acquired immune system lymphocytes, T lymphocytes and B lymphocytes, and also in innate immune system lymphocytes. Can be the subject of treatment with the pharmaceutical composition of the present invention.
  • the inflammatory disease can be a disease or condition that is resistant to steroids or resistant to treatment that blocks the binding of IL-7R to IL-7.
  • a method of treating cancer or an inflammatory disease in a subject in need thereof comprising the step of administering to the subject an antibody drug conjugate of an anti-IL-7R antibody or antigen-binding fragment thereof and a cytotoxic agent.
  • a method is provided that includes administering a gate. The conjugate can be administered in a therapeutically effective amount.
  • the subject can be a subject that is resistant to steroid treatment or has received steroid treatment.
  • a method of treating cancer or an inflammatory disease in a subject in need thereof comprising the step of administering an antibody drug conjugate of an anti-IL-7R antibody or antigen-binding fragment thereof to a cytotoxic agent.
  • a method is provided comprising administering a gate and administering a steroidal anti-inflammatory drug to the subject.
  • the subject can be a subject that is resistant to steroid treatment or has received steroid treatment.
  • a method includes administering a conjugate.
  • a method of reducing steroid resistance of a disease or condition in a subject in need thereof, the subject comprising an antibody drug of an anti-IL-7R antibody or antigen-binding fragment thereof and a cytotoxic agent includes administering a conjugate.
  • a medicament for use in treating cancer or an inflammatory disease comprising an antibody drug conjugate of an anti-IL-7R antibody or antigen-binding fragment thereof and a cytotoxic agent.
  • the use of anti-IL-7R antibodies is provided.
  • in the manufacture of a medicament for use in treating cancer or an inflammatory disease comprising an antibody drug conjugate of an anti-IL-7R antibody or antigen-binding fragment thereof and a cytotoxic agent.
  • Use of an anti-IL-7R antibody or antigen-binding fragment thereof and a cytotoxic agent is provided.
  • an antibody drug conjugate of an anti-IL-7R antibody or antigen-binding fragment thereof and a cytotoxic agent in the manufacture of a medicament for use in treating cancer or an inflammatory disease.
  • One aspect of the invention provides the use of an antibody drug conjugate of an anti-IL-7R antibody or antigen-binding fragment thereof and a cytotoxic agent in a method for treating cancer or an inflammatory disease. Is done.
  • IL-7R expression may be suppressed in IL-7R-expressing cells.
  • a pharmaceutical composition for use in treating cancer or inflammatory disease comprising an IL-7R expression inhibitor.
  • the present invention also provides a pharmaceutical composition for use in treating a steroid resistant disease or condition, comprising an IL-7R expression inhibitor.
  • the present invention further provides a pharmaceutical composition for use in reducing steroid resistance in a disease or condition, comprising an IL-7R expression inhibitor.
  • the present invention further provides a pharmaceutical composition for use in treating a steroid resistant disease or condition, comprising an IL-7R expression inhibitor used in combination with a steroid.
  • the present invention further provides a pharmaceutical composition for use in suppressing lymphadenopathy in a cancer subject, comprising an IL-7R expression inhibitor.
  • IL-7R expression inhibitors include siRNA, shRNA and antisense oligos against IL-7R, which can be used in the present invention.
  • examples of the IL-7R expression inhibitor include an expression vector containing a DNA encoding siRNA or shRNA.
  • Example 1 Survival activity of IL-7 receptor and tumor cells
  • IL-7 receptor IL-7 receptor
  • IL-7R IL-7 receptor
  • IL-7R expression was measured by flow cytometry using an anti-human IL-7R antibody (eBioRDR5, eBioscience; R34-R34, TONB0) and an anti-mouse IL-7R antibody (A7R34 or SB / 199, eBioscience).
  • IL-7R antibody (A7R34) was prepared as in References 2 and 3 below and used in this example. The result was as shown in FIG. 1A.
  • IL-7R was positive in human T cell malignant lymphoma cell line SupT1, human B cell lymphocytic leukemia cell line NALM6, and mouse lymphocytic leukemia cell line CYG82.
  • lung cancer lymph node metastasis derived cell line H2009 lung cancer alveolar metastasis derived cell line H358, lung cancer pleural effusion cell line LC-2 / ad, pancreatic cancer liver metastasis derived cell line PK-45H, pharyngeal cancer IL-7R was positive in the cancerous pleural effusion cell line Detroite 562, prostate cancer bone metastasis-derived cell line PC3, and bladder cancer cell line TCCsup with bone metastasis (the histogram on the right side of each figure shows IL- 7R expression, left histogram is negative control).
  • IL-7R was positive in all cell lines of lymphoid malignancies and metastatic solid tumors examined. Expression was confirmed in metastatic solid tumors
  • IL-7R is expressed by expressing short hairpin RNA (shRNA, Sigma) incorporated into a lentiviral vector.
  • a gene knockdown (KD) cell line (IL-7R-KD) was generated.
  • a control cell (CTR) for comparison was prepared by introducing shRNA (Sigma) of the GFP gene into SupT1, NALM6, and CYG82, and non-specific shRNA (Sigma) into H2009 and PK-45H.
  • CDNA was synthesized from the total RNA of the cells using an extraction kit (QIAGEN) and a synthesis kit (Thermo Fisher Scientific or Takara).
  • Quantitative RT-PCR was performed. That is, ABI7500Fast (Thermo Fisher Scientific) was used in a total of 20 ⁇ l of the reaction mixture of 9 ⁇ l of cDNA dilution, 1 ⁇ l TaqMan primers / probe mixture (Thermo Fisher Scientific) and 10 ⁇ l TaqMan Fast Universal PCR Master Mix (Thermo Fisher Scientific). ). The expression of each gene was standardized by the comparative CT method using GAPDH for human genes and ACTb for mouse genes.
  • Example 2 Physiological function of IL-7R In this example, further physiological function of IL-7R was examined.
  • cytotoxic effect (IC50; 50% cytotoxic concentration) of dexamethasone in each cell in vitro was examined. confirmed. 1000 to 3000 cells were seeded at 100 ⁇ L / well in a 96-well plate, and dexamethasone was added to each well at a concentration of 0.1 ⁇ M to 1 mM.
  • WST-8 (Dojindo, Cell-Counting-Kit-8) was added at 10 ⁇ L / well, and after culturing for 3 hours, absorbance at 450 nm was measured with a microplate reader (Molecular Devices). IC50 was calculated by calculating the cell viability at each concentration. The result was as shown in FIG. 2A.
  • IL-7R-KD cells As shown in FIG. 2A, in SupT1, NALM6, and CYG82, the IC50 of steroid (dexamethasone) was decreased and the steroid sensitivity was increased in IL-7R-KD cells. . In H2009 and PK45H, there was no difference between IL-7R-KD cells and CTR cells. Thus, it is clear that IL-7R is greatly involved in steroid sensitivity in lymphoid malignant tumors, whereas in metastatic solid tumors H2009 and PK45H, steroid sensitivity is It became clear that IL-7R was hardly involved.
  • a steroid resistant strain was obtained, and an IL-7R knockdown strain and a control strain (CTR) with a specific shRNA were prepared to confirm the effect of IL-7R knockdown in the presence of dexamethasone. did. Since a CYG82 steroid resistant strain can be easily obtained from the CYG82 parental strain, the growth of cells in a state where a high concentration of steroid was added was observed. 10 6 cells were seeded at 3 ml / well in a 6- well plate, 10 nM or 100 nM dexamethasone was added, and the number of cells after 1 week and 2 weeks was counted. The result was as shown in FIG. 2B.
  • cDNA is synthesized from total RNA of cells using an extraction kit (QIAGEN) and a synthesis kit (Thermo Fisher Fisher or Takara), quantitative PCR is performed, and the expression of IL-7R gene and CD19 gene is performed. Examined. The result was as shown in FIG. 2C.
  • the steroid resistant strain of CYG82 cells obtained above shows a high IC50 against dexamethasone. Then, when the expression level of IL-7R was compared between the non-resistant parent strain and the resistant strain, it was revealed that the expression of IL-7R was significantly improved in the resistant strain. On the other hand, regarding the expression level of CD19, no significant difference was observed between the parent strain and the resistant strain.
  • Example 3 Relationship between IL-7R Signal Activation and Steroid Resistance
  • the relationship between IL-7R signal activation and steroid resistance was examined.
  • JAK / STAT was measured by fixing cells with 4% paraformaldehyde, treating with methanol, and detecting phosphorylated STAT5 using an anti-phospho-STAT5 antibody (eBioscience). Measurement was performed using a flow cytometry detection device, Guava easyCyte 10HT (Merck Millipore) or Aria flow cytometer (BD Biosciences). Dead cells were stained with Propidium iodide (Thermo Fisher Scientific) and excluded during measurement.
  • FlowJo program (Tree Star) was used for data analysis.
  • the BCL2 signal was subjected to total RNA extraction and cDNA synthesis from each cell, and the expression level of BCL2 was examined by quantitative PCR.
  • For NF ⁇ , cells were fixed with 4% paraformaldehyde and reacted with anti-phospho-p65 / NF-k ⁇ antibody (Abcam).
  • As the secondary antibody Alexa-488 / 555 / 647-labeled anti-rabbit IgG antibody (Thermo Fisher Scientific) was used. Nuclei were stained with DAPI (Thermo Fisher Scientific).
  • NFkB was decreased by IL-7R knockdown in all cells, but also decreased in IL-7R-KD of H2009 and PK-45H cells. Since these cells had no relationship with steroid resistance as shown in FIG. 2A, NFkB seemed to be more likely to be more strongly involved in cell proliferation and survival than steroid resistance.
  • Example 4 Acquisition of steroid resistance and change in gene expression
  • the relationship between IL-7R gene expression and steroid resistance in clinical samples was examined.
  • GSE39339 and GSE32962 were obtained from Gene® Expression® Omnibus® (GEO), and clinical data of E-MEXP-3916 was also acquired from Array® Express.
  • the data was subjected to statistical analysis after being standardized by RMA processing using analysis software R (open source freeware).
  • RMA analysis software
  • SPSS Statistics Version 20 SPSS Statistics Version 20 (IBM) was used, and P ⁇ 0.05 or less was used as a criterion.
  • FIG. 4A shows the analysis result of GSE39339
  • FIG. 4B shows the analysis result of GSE32962
  • FIG. 4C shows the analysis result of E-MEXP-3916.
  • lymphocyte markers TdT, CD19, and CD22 significantly decreased after chemotherapy including steroid (dexamethasone) for human acute lymphoblastic leukemia.
  • steroid dexamethasone
  • IL-7R tended to increase after treatment.
  • Chemotherapy included anthracycline, vincristine, and L-asparakinase in addition to dexamethasone.
  • a steroid (prednisolone) sensitive and resistant strain was established from cells derived from a patient with human acute B lymphoblastic leukemia and subjected to DNA microarray analysis. There was no change in TdT, CD19, and CD22 between the two groups, but IL-7R was significantly higher in the steroid (prednisolone) resistant strain.
  • DNA microarray analysis was performed in a non-treated group and a treated group containing steroid (dexamethasone) in a patient with human acute T lymphocytic leukemia.
  • IL-7R expression was significantly higher in the treatment group.
  • Chemotherapy included daunorubicin, vincristine, and L-asparakinase in addition to dexamethasone.
  • Example 5 Characteristics of anti-IL-7R antibody
  • the kinetics when an anti-IL-7R antibody was brought into contact with cells were examined.
  • Alexa647 (Thermo Fisher Scientific) was labeled with an anti-IL-7R antibody (A7R34). Alexa647-labeled anti-IL-7R antibody (A7R34) was administered to CYG62 cells under in vitro culture conditions, and intracellular uptake after 10 and 30 minutes was observed. Lysosomes were visualized with a lyso tracker (Thermo Fisher Fisher Scientific). The result was as shown in FIG. 5A.
  • the anti-IL-7R antibody was taken up into cells within 30 minutes after administration and localized in lysosomes.
  • the kinetics of anti-IL-7R antibody in vivo was confirmed. Specifically, after the CYG82 parent strain was formed into a tumor after subcutaneous implantation of female nude mice, Alexa647-labeled anti-IL-7R antibody (A7R34) or control antibody was administered from the tail vein of the mouse. One week after administration, the tumor, lymph node, spleen, bone, and liver were removed, and accumulation of anti-IL-7R antibody (A7R34) in each tissue was confirmed. The result was as shown in FIG. 5B.
  • the anti-IL-7R antibody (A7R34) accumulated in the tumor and the diseased lymph node, whereas the control antibody did not accumulate in these tissues.
  • CYG82 cells and IL-7-dependent RAG2 ⁇ / ⁇ cells were added to each 96-well plate at 2000 or 4000 cells / 100 ⁇ L / well and cultured.
  • IL-7R antibody A7R34 was added in a dilution series of 10 ⁇ 4 to 10 2 ⁇ g / ml, and 72 hours later, WST-8 (Dojindo, Cell Counting Kit-8) was added at 10 ⁇ L / well and cultured for 3 hours. Later, the absorbance at 450 nm was measured with a microplate reader (Molecular Devices). The cell viability was calculated and the cell killing effect due to the neutralizing activity of IL-7 signal of the antibody was measured. The result was as shown in FIG. 5C.
  • the anti-IL-7R antibody (A7R34) showed a cell-killing effect due to neutralizing activity in IL-7-dependent RAG2 ⁇ / ⁇ cells, but almost no effect in CYG82 cells. I could't. This revealed that CYG82 cells do not require IL-7. In CYG82 cells, cell proliferation and survival are maintained by autoactivation of IL-7R, and it is considered that the influence of the neutralizing activity of IL-7R antibody (A7R34) is avoided.
  • the linker and drug use maleimide for binding to the antibody.
  • Mal-PEG 12 -OSu was purchased from Quanta Biodesign or IRIS Biotech GMBH and used to increase polarity.
  • a carbamate bond was introduced into the linker site as a cleavage site for releasing the drug inside the cell.
  • SN-38 was dissolved in dimethyl sulfoxide at a concentration of 10 mM and stored at ⁇ 80 ° C.
  • the ADC was prepared by reducing the SS bond in the antibody and reacting the maleimide group of the linker with the drug. It was possible to obtain an ADC in which 4 to 6 drugs were bound per antibody. The result was as shown in FIG. 5D.
  • A7R34-ADC (SN-38) was 100 times more potent than CPT-11 and showed a cell killing effect similar to free SN-38. This indicates that A7R34-ADC is effective against CYG82 that survives independent of IL-7. This indicates that IL-7R constitutively internalizes even in the absence of IL-7 signal.
  • the anti-IL-7R antibody (A7R34)
  • the neutralizing activity between IL-7 and IL-7R did not significantly affect the cell viability and tumor volume suppression effects.
  • a remarkable antitumor effect was exhibited by the ADC of the anti-IL-7R antibody and the cytotoxic agent.
  • Example 6 Anti-tumor effect in leukemia model
  • the anti-tumor effect by ADC of anti-IL-7R antibody and drug was confirmed in leukemia model.
  • a mouse having a tumor volume of about 50 mm 3 after subcutaneous implantation of female nude mice (5 mice per group) with the CYG82 parent strain was used as a leukemia model mouse.
  • A7R34-ADC (SN-38) prepared in Example 5 was used as the ADC.
  • A7R34-ADC (SN-38) was administered as an ADC amount of 50 mg / kg on days 0, 4 and 8 from the tail vein of the mouse, and 0.6 mg / kg as an SN-38 equivalent amount.
  • Tumor volume was measured at 2-day intervals and measured until day 26 after administration.
  • 50 mg / kg of A7R34-ADC [0.6 mg / kg in terms of SN-38] was transferred to the mouse via the tail vein. Administered on day 8.
  • ADC of anti-IL-7R antibody and drug showed the most excellent inhibitory effect on tumor volume increase. On the other hand, no significant effect was observed with the anti-IL-7R antibody alone. Further, as shown in FIG. 6B, the ADC of the anti-IL-7R antibody and the drug showed a strong antitumor effect even against a steroid-resistant lymphocytic leukemia model.
  • Example 7 Lymph node enlargement suppression CYG82 cells in a leukemia model were transplanted into the front sole of female nude mice (5 mice per group), and the average volume of lymph nodes was measured on the 16th day after transplantation. The result was as shown in FIG. 7A. As shown in FIG. 7A, the enlargement of axillary lymph nodes was markedly suppressed in IL-7R knockdown mice.
  • A7R34-ADC (SN-38) was administered on days 0, 4 and 8 from the mouse tail vein.
  • physiological saline, A7R34 antibody, isotype antibody ADC (SN-38), dexamethasone, or CPT-11 was administered.
  • the volume of the lymph nodes was measured. The result was as shown in FIG. 7B.
  • A7R34-ADC (SN-38) showed a strong lymph node enlargement suppressing effect. Almost no therapeutic effect was observed with A7R34 monotherapy.
  • Example 8 Radioisotope-labeled anti-IL-7R antibody
  • ADC radioisotope-labeled anti-IL-7R antibody
  • the preparation of 90 Y-labeled antibody was performed according to Reference Document 10 below.
  • 1M acetate buffer (pH 6.0) and 90 YCl 3 were mixed at 1: 1 and allowed to stand at room temperature for 5 minutes.
  • a commercially available anti-IL-7R antibody (clone 40131, R & D) was added and allowed to stand at room temperature for 30 minutes.
  • purification was performed using a Sephadex G50 (GE Healthcare) column (swelled with 0.1 M acetate buffer (pH 6.0)). Thereafter, the dose was adjusted to 20 ⁇ g in terms of unlabeled anti-human IL-7R antibody.
  • 90 Y-labeled anti-IL-7R antibody was administered to the tail vein of nude mice transplanted subcutaneously with either SupT1 or H2009 to confirm the drug efficacy.
  • a single dose of 90 ⁇ g of 90 Y-labeled anti-IL-7R antibody was administered. The results were as shown in FIGS. 8A and B.
  • FIGS. 8A and 8B it was revealed that at 90 ⁇ Ci, 90 Y-labeled anti-IL-7R antibody exhibits an anti-tumor effect. This revealed that the anti-IL-7R antibody can be used for radioimmunotherapy.
  • Example 9 Effect of ADC on immune response
  • the effect of ADC on tumor was confirmed.
  • mice Healthy female BALB / c mice receive saline, 50 mg / kg A7R34-ADC (SN-38) [0.6 mg / kg in SN-38 equivalent], 10 mg / kg dexamethasone, or 40 mg / kg dexamethasone. Each was administered from the tail vein of mice.
  • IL-7R positive T cells and B cells were reduced to 1/10 or less in any of the thymus, bone marrow, and spleen.
  • T cells including IL-7R positive T cells in the thymus were generally decreased, but conversely, IL-7R positive B cells in the bone marrow increased more than twice.
  • the ratio of IL-7R positive T cells is increased, and physiologically IL-7R is involved in steroid resistance of T cells and B cells. It was shown that.
  • A7R34-ADC is considered effective in controlling steroid-resistant IL-7R positive cells.
  • Example 10 Increase in IL-7R positive cells in immune diseases
  • IL-7R gene expression in clinical samples was examined in the same manner as in Example 4.
  • Raw data of GSE55235 and GSE14580 were obtained from Gene Expression Omnibus (GEO). Data was analyzed using analysis software Qlucore Omics Explorer (Filgen), and P ⁇ 0.05 or less was used as a criterion for judgment. The result was as shown in FIG.
  • IL-7R gene expression tended to be higher in normal rheumatoid arthritis (RA) patients than in healthy subjects.
  • RA normal rheumatoid arthritis
  • U ulcerative colitis
  • IL-7R gene expression tended to be higher than that in healthy subjects, and in particular, IL-7R gene expression was enhanced in patients who did not succeed in both anti-TNF.
  • Example 11 Therapeutic effect of rheumatoid arthritis model using anti-IL-7R antibody
  • the anti-IL-7R antibody-drug conjugate was administered to a rheumatoid arthritis model, and the effect was examined.
  • A7R34-ADC As a conjugate of the anti-IL-7R antibody and the drug, A7R34-ADC (MMAE) in which A7R34 monoclonal antibody and monomethyl auristatin E (MMAE) are linked by Val-Cit bond (valine-citrulline dipeptide) was used. The Val-Cit bond and MMAE were linked by p-aminobenzylcarbamate (PABC). MMAE is known to have stronger cytotoxic activity than SN-38.
  • A7R34-ADC (MMAE) was prepared as described in References 6 to 9 as in Example 5. The chemical structural formula of A7R34-ADC (MMAE) was as follows.
  • a rheumatoid arthritis model was prepared by the method described in Reference 7 below. That is, 2 mg of anti-collagen 4 antibody (Chondrex) was intraperitoneally administered to female DBA / 1J mice (day 0). On day 3, 50 ⁇ g of LPS (Chondrex) was administered intraperitoneally. On the 5th day, 50 mg / kg of A7R34-ADC (MMAE) [MMAE equivalent 1 mg / kg] or 50 mg / kg of A7R34-ADC (SN-38) [SN-38 equivalent] The dose was 0.6 mg / kg].
  • MMAE A7R34-ADC
  • SN-38 A7R34-ADC
  • physiological saline 1 mg / kg MMAE, 50 mg / kg isotype control antibody ADC (MMAE) [1 mg / kg in MMAE equivalent amount], 10 mg / kg dexamethasone, or 50 mg / kg anti-TNF preparation etanercept was administered instead.
  • MMAE isotype control antibody ADC
  • SN-38 physiological saline as a control
  • 50 mg / kg anti-IL-7R antibody (A7R34) 50 mg / Kg anti-CD19 antibody ADC
  • SN-38 10 mg / kg dexamethasone was used.
  • the severity of arthritis was evaluated by measuring the arthritis score of each individual according to the instructions (the higher the score is, the more severe it is). The results were as shown in FIGS. 11A-C.
  • the arthritis score was determined by observing three joints: fingers, back and wrists of each limb. Specifically, the arthritis score of each limb was determined based on the following criteria. Evaluation criteria score of arthritis score of each limb 0: When there is no inflammation in any joint, Score 1: If inflammation is observed in one of the joints, Score 2: If inflammation is observed in any two of the joints, Score 3: If inflammation is observed in all joints, Score 4: All joints are inflamed and the entire limb is swollen red. Based on the above scoring, the arthritis score was measured for each limb (4 points maximum for each limb), and the limbs were summed (16 points maximum) to determine the arthritis score of the individual.
  • A7R34-ADC remarkably suppressed the exacerbation of the arthritic coefficient in the rheumatoid arthritis model, like dexamethasone.
  • the dexamethasone-administered group there was an individual (an example of poor control of inflammation) in which inflammation in the forelimbs was marked compared to the hindlimbs.
  • the A7R34-ADC (MMAE) administration group both the forelimbs and hindlimbs showed stable and strong anti-inflammatory effects. This anti-inflammatory effect was superior to anti-TNF therapy with etanercept.
  • FIG. 11B Similar results were obtained with A7R34-ADC (SN-38).
  • anti-IL-7R antibody A7R34 antibody alone was observed to suppress the deterioration of arthritis coefficient in a rheumatoid arthritis model, but the effect was limited (see FIG. 11B).
  • FIG. 11C a tissue section of a mouse forefoot lesion on day 9 after drug administration was prepared and observed with hematoxylin and eosin staining (HE staining). As shown in FIG. 11C, it can be seen that inflammation was significantly improved in the ADC administration group.
  • Example 12 Treatment effect of ulcerative colitis using anti-IL-7R antibody
  • the effect of anti-IL-7R antibody on ulcerative colitis was examined.
  • the ulcerative colitis model was prepared as shown in Reference Material 11 below. That is, female BALB / c mice were allowed to drink 5% dextran sulfate (DSS; MP Biomedicals). From the second day after drinking DSS, A7R34-ADC (MMAE) or A7R34-ADC (SN-38) was intraperitoneally administered in the same manner and dose as in the rheumatoid arthritis model.
  • DSS dextran sulfate
  • MMAE isotype control antibody ADC
  • dexamethasone dexamethasone
  • anti-TNF preparation etanercept was administered instead of ADC.
  • A7R34-ADC physiological saline, anti-IL-7R antibody (A7R34), anti-CD19 antibody ADC (SN-38), and dexamethasone were used as controls as in the rheumatoid arthritis model.
  • the large intestine on the 8th day after drinking was removed and the total length was measured. Furthermore, a tissue specimen was prepared and a pathological image was confirmed by HE staining.
  • the severity of ulcerative colitis was observed in terms of weight loss (the greater the weight loss, the greater the severity), the entire length of the large intestine (the more severe the length of the large intestine), and histological changes.
  • the results of changes in body weight were as shown in FIGS. 12A and B. Moreover, the result of the histological change of the entire length of the large intestine was as shown in FIG.
  • A7R34-ADC significantly suppressed weight loss after the fifth day.
  • This inhibitory effect was significantly higher than the suppression of weight loss in the etanercept administration group.
  • MMAE and control antibody ADC showed almost no inhibitory effect on weight loss.
  • A7R34-ADC also remarkably suppressed weight loss in model mice.
  • FIG. 13 in the negative control for ulcerative colitis, the total length was significantly reduced compared to the DSS-untreated group, but in the A7R34-ADC (MMAE) administration group, the total length of the large intestine was DSS. The laparotomy was at the same level as the untreated group. In the A7R34-ADC (SN-38) administration group, the total length tended to be long.
  • IL-7R is expressed on acquired immune system lymphocytes, T cells and B cells, and in particular on innate immune system lymphocytes. Since the inflammation could be sedated by the ADC against IL-7R, the ADC is effective against the sedation of inflammation via the acquired immune system lymphocytes, T cells and B cells, and in particular the innate immune system lymphocytes. It was suggested to be effective. Therefore, according to the present invention, against all inflammation via innate immune system lymphocytes, diseases caused by enhancement of acquired immune system (for example, autoimmune diseases, graft-versus-host diseases, etc.) and other diseases or conditions described above It has been shown that antibody drug conjugates of anti-IL-7R antibodies and cytotoxic agents are effective.

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Abstract

La présente invention concerne : un conjugué d'anticorps-médicament d'un anticorps anti-IL-7R ; et une composition pharmaceutique de traitement du cancer ou de l'inflammation et contenant un conjugué d'anticorps-médicament d'un anticorps anti-IL-7R et d'un médicament cytotoxique. La présente invention fournit : un conjugué d'anticorps-médicament d'un anticorps anti-IL-7R et d'un médicament cytotoxique ; et une composition pharmaceutique pour l'utilisation dans le traitement du cancer ou d'une inflammation, et contenant un conjugué d'anticorps-médicament d'un anticorps anti-IL-7R et d'un médicament cytotoxique.
PCT/JP2018/007985 2017-03-03 2018-03-02 Conjugué d'anticorps-médicament de l'anticorps anti-il-7r, et composition pharmaceutique de traitement du cancer ou de l'inflammation et contenant un conjugué d'anticorps-médicament de l'anticorps anti-il-7r et d'un médicament cytotoxique Ceased WO2018159808A1 (fr)

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JP2019503147A JPWO2018159808A1 (ja) 2017-03-03 2018-03-02 抗ｉｌ−７ｒ抗体の抗体薬物コンジュゲートと、がんまたは炎症を処置することに用いるための、抗ｉｌ−７ｒ抗体と細胞傷害剤との抗体薬物コンジュゲートを含む医薬組成物

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JP2017-040691 2017-03-03
JP2017040691 2017-03-03

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WO2025089399A1 (fr) * 2023-10-26 2025-05-01 国立研究開発法人国立がん研究センター Anticorps dirigé contre le récepteur de l'interleukine-7 (il-7r), et conjugué anticorps-médicament (adc) comprenant ledit anticorps

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JP2009102367A (ja) * 2001-04-06 2009-05-14 Univ Of Bristol ステロイド耐性患者におけるｃｄ２５結合分子の使用
JP2012024095A (ja) * 2004-08-10 2012-02-09 Cardiff Biologicals Ltd 乳癌の予後診断方法およびキット
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JP2009102367A (ja) * 2001-04-06 2009-05-14 Univ Of Bristol ステロイド耐性患者におけるｃｄ２５結合分子の使用
JP2012024095A (ja) * 2004-08-10 2012-02-09 Cardiff Biologicals Ltd 乳癌の予後診断方法およびキット
JP2014012657A (ja) * 2007-03-20 2014-01-23 Clavis Pharma Asa 抗炎症剤及び抗癌剤であるグルココルチコイドの脂肪酸エステル
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2025089399A1 (fr) * 2023-10-26 2025-05-01 国立研究開発法人国立がん研究センター Anticorps dirigé contre le récepteur de l'interleukine-7 (il-7r), et conjugué anticorps-médicament (adc) comprenant ledit anticorps

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TW201834698A (zh) 2018-10-01

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