WO2011071099A1 - Anticorps anti-peptide de mortaline anticancéreux - Google Patents

Anticorps anti-peptide de mortaline anticancéreux Download PDF

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WO2011071099A1
WO2011071099A1 PCT/JP2010/072086 JP2010072086W WO2011071099A1 WO 2011071099 A1 WO2011071099 A1 WO 2011071099A1 JP 2010072086 W JP2010072086 W JP 2010072086W WO 2011071099 A1 WO2011071099 A1 WO 2011071099A1
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mortalin
peptide
seq
amino acid
antibody
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PCT/JP2010/072086
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ワダワ　レヌー
スニル カウル
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独立行政法人産業技術総合研究所
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Priority to EP10836019.9A priority Critical patent/EP2511371B1/fr
Priority to US13/514,755 priority patent/US8586042B2/en
Priority to JP2011545235A priority patent/JP5641486B2/ja
Publication of WO2011071099A1 publication Critical patent/WO2011071099A1/fr

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    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/04—Immunostimulants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • 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
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/30—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants from tumour cells
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G01N33/57407—Specifically defined cancers
    • G01N33/57415—Specifically defined cancers of breast
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G01N33/57407—Specifically defined cancers
    • G01N33/57423—Specifically defined cancers of lung
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
    • C07K2317/34—Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/76—Antagonist effect on antigen, e.g. neutralization or inhibition of binding

Definitions

  • the present invention relates to an anti-mortalin peptide antibody exhibiting high anticancer activity and the antibody-producing hybridoma.
  • Mortalin (mortalin 2) belongs to the Hsp family of heat shock proteins and is a heat non-responsive protein, but binds to p53, a tumor suppressor protein, and inactivates its transcriptional activity function (Non-patent Document 1). The action and the like have been gradually found, and it has been clarified that it is essentially involved in carcinogenesis (Patent Document 1, etc.). Recently, research and development on substances that suppress mortalin and its functions and functions have been activated, and mortalin antibodies that bind to mortalin are also highly expected as anticancer agents (Non-patent Documents 2-6). .
  • Patent Document 1 filed an application regarding a pharmaceutical composition for cancer treatment using the antibody, a drug carrier, etc., and further examined the epitope sequence of mortalin recognized by the anti-mortalin antibody in detail, Several types of common epitopes including the epitope sequence “LFGRAP” that are commonly recognized by anti-mortalin antibodies having an activating function were determined (Patent Document 2).
  • anti-mortalin antibodies that have internalization functions in cancer cells have anti-cancer effects
  • anti-mortalin monoclonal antibodies using the peptide containing the common epitope as an immunogen, an even stronger anti-cancer effect can be obtained. It has been expected that an anti-mortalin peptide antibody will be obtained, but the present situation is that a peptide antibody having an anticancer activity exceeding that of the known antibody has not yet been provided.
  • Wadhwa, R., Takano, S., Robert, M., Yoshida, A., Reddel, R., Nomura, H., Mitsui, Y., and Kaul, S. C. (1998) J Biol Chem 273, 29586-29591 Walker, C., Bottger, S., and Low, B. (2006) Am J Pathol 168, 1526-1530 Wadhwa, R., Sugihara, T., Yoshida, A., Nomura, H., Reddel, R. R., Simpson, R., Maruta, H., and Kaul, S. C.
  • the present invention is a peptide antibody having a peptide containing an epitope sequence that is commonly recognized by an anti-mortalin antibody having an internalization function in cancer cells as an immunogen, and more than an anti-mortalin antibody having an internalization function in cancer cells.
  • An object of the present invention is to provide an anti-mortalin peptide antibody having a high anticancer activity.
  • Another object of the present invention is to provide an anticancer agent having a tumor cell growth inhibitory effect using the antibody.
  • the present inventors represent the epitope “LFGRAP” (sequence 402-407, position from the N-terminal of the human mortalin amino acid sequence as a common recognition site of anti-mortalin antibodies having an internalizing function in cancer cells. The same applies hereinafter. ) was designed, and at the same time, focusing on other common epitope “KAMQDAEVSKSDIGEVI” (sequence 368-384), an 18 amino acid peptide containing the sequence was designed. Both peptides were used together as an immunogen cocktail.
  • mice were immunized with the immunogen cocktail, a large number of monoclonal antibody-producing hybridomas were prepared by a conventional method, and 8 clones were selected by specificity for mortalin antigen and internalization into cells. Furthermore, as a result of examining the cancer cell growth inhibitory effect of the antibodies produced by these clones, it was found that only the monoclonal antibodies produced by C-26 and C-69 clones markedly inhibit the growth of cancer cells. The inhibitory effect far exceeded that of anti-mortalin monoclonal antibodies having a known internalization function. Obtaining the above knowledge, the present invention has been completed.
  • the hybridomas producing the C-26 antibody and the C-69 antibody were deposited as FERM P-21875 and FERM P-21876, respectively, at the National Institute of Advanced Industrial Science and Technology Patent Microorganism Depositary.
  • the present invention is specifically as follows.
  • a hybridoma obtained using a peptide shown in SEQ ID NO: 4 and a 1: 1 immunogen cocktail of the peptide shown in SEQ ID NO: 6 as an immunogen, specifically recognizing human mortalin antigen and internalizing cancer cells A hybridoma strain C-26 (FERM P-21875) or C-69 strain (FERM P-21875) having the ability to produce a functional monoclonal antibody.
  • a monoclonal antibody that specifically recognizes the human mortalin antigen produced by the hybridoma strain of [1] above and has a function of internalizing cancer cells, or a fragment containing the antigen-binding site thereof.
  • An anticancer agent having a tumor cell growth inhibitory effect comprising the monoclonal antibody according to [2] or a fragment containing an antigen-binding site thereof as an active ingredient.
  • a reagent for detecting a tumor cell comprising using the monoclonal antibody according to [2] or a fragment containing an antigen-binding site thereof.
  • the peptide of the following (1) or (2) which functions as an epitope recognized by a human mortalin-specific monoclonal antibody having a cancer cell internalization function; (1) a peptide comprising an amino acid sequence represented by SEQ ID NO: 10 or an amino acid sequence containing at least a partial sequence represented by SEQ ID NO: 8 in the sequence; (2) A peptide comprising an amino acid sequence represented by SEQ ID NO: 11 or an amino acid sequence containing at least a partial sequence represented by SEQ ID NO: 9 in the sequence.
  • An epitope set for an immune cocktail for producing a human mortalin-specific monoclonal antibody having a cancer cell internalizing function characterized by combining the following peptides (1) and (2): (1) a peptide comprising an amino acid sequence represented by SEQ ID NO: 10 or an amino acid sequence containing at least a partial sequence represented by SEQ ID NO: 8 in the sequence; (2) A peptide comprising an amino acid sequence represented by SEQ ID NO: 11 or an amino acid sequence containing at least a partial sequence represented by SEQ ID NO: 9 in the sequence.
  • an anti-mortalin peptide antibody having an extremely excellent anticancer activity could be provided.
  • the anti-cancer activity of the peptide antibody was compared with the 37-6 antibody that showed the highest anti-cancer activity among the monoclonal antibodies having the internalizing function of cancer cells obtained using full-length mortalin as an immunogen.
  • the effect of inhibiting cancer cell growth is several tens of times higher.
  • the anticancer agent which has the outstanding cancer cell growth inhibitory activity can be provided by making the said antibody into an active ingredient.
  • the monoclonal antibody of the present invention retains the internalization function of cancer cells, it can be used as a cancer cell detection agent by labeling.
  • a peptide containing two common epitopes used to produce peptide antibodies Confirmation of specificity to antigen: screening of hybridomas by Western blotting.
  • R indicates the result in the recombinant protein extract
  • U indicates the result in the human cancer cell (U2OS) solution.
  • the degree of reactivity was indicated by the number of +.
  • eight selected clones are shown.
  • Confirmation of specificity for antigen Screening of human cancer cells by immunostaining. The degree of reactivity is indicated by the number of +. In the lower left frame, eight selected clones are shown. Confirmation of cellular uptake.
  • a hybridoma is added to the culture solution of human cancer cells, and the uptake into cells is screened by immunostaining. The degree of cellular uptake was indicated by the number +. In the lower left frame, eight selected clones are shown. Figures 2-4 are shown together. Anticancer activity of monoclonal antibody produced by C-26 clone. Anticancer activity of monoclonal antibody produced by C-67 clone. Anticancer effect of monoclonal antibody produced by C-69 clone. Anticancer activity of monoclonal antibody produced by C-131 clone. Anti-cancer effect of monoclonal antibody produced by C-133 clone. Anti-cancer effect of monoclonal antibody produced by C-137 clone.
  • Anticancer activity of monoclonal antibody produced by C-152 clone Anticancer effect of monoclonal antibody produced by C-177 clone. Comparison of anticancer activity of monoclonal antibodies produced by 8 hybridomas. Amino acid sequences of 36 types of peptides and epitopes recognized by C26 and C69 antibodies. Epitope analysis of antibodies. Array analysis of 15 amino acid peptides shifted by one amino acid for the mortalin amino acid sequence 368-417. The vertical axis represents the signal intensity in ELIZA. Comparison of anticancer activity between C26 and C69 antibodies and other antigen peptide antibodies.
  • C133 is a monoclonal antibody with the same immunogen but no epitope in residues 368 to 417
  • 37-6 is a monoclonal antibody that recognizes the “LFGRAP” site with the entire mortalin as the immunogen (patented) Reference 1).
  • mortalin usually refers to human mortalin.
  • the homology between mot-2) and human mortalin is 97.9% at the amino acid level (BLAST method). Since the epitope sequences are also the same, they may be derived from other species such as mouse mortalin.
  • NM_010481 human mortalin: AK315177 , SEQ ID NO: 1
  • the present inventors have already reported that a mortalin antibody having the ability to internalize cells has an anticancer effect (Patent Document 1, etc.).
  • the epitope region that is recognized in common by the mortalin antibody having the ability to internalize cells is found to exist in a region corresponding to positions 381 to 410 of human mortalin in the amino acid sequence of human mortalin, and 348 including positions 381 to 410 are included.
  • the consensus sequence of the binding sequence specific for the internalizing antibody was “LFGRAP (SEQ ID NO: 2)” (common epitope).
  • LFGRAP consensus sequence of the binding sequence specific for the internalizing antibody
  • a plurality of binding sequences specific to other internalizing antibodies were found, one of which is “KAMQDAEVSKSDIGEVI (SEQ ID NO: 3)” (Patent Document 2).
  • peptide antibody “QDLFGRAPSKAVNPDEA (SEQ ID NO: 5)” having the highest calculated antigenic score among 17 amino acids including “KAMQDAEVSKSDIGEVI (SEQ ID NO: 3)” and “LFGRAP (SEQ ID NO: 2)”
  • the peptide-1 (SEQ ID NO: 4) and peptide-2 (SEQ ID NO: 6) with cysteine (C) added to the carrier protein at each end thereof were immunized with the carrier protein at a ratio of 1: 1.
  • a monoclonal antibody-producing hybridoma was obtained by a conventional method.
  • hybridoma clones by confirming cancer cell growth inhibitory activity of peptide antibodies (1) Screening of hybridomas by Western blotting (2) Screening by immunostaining of human cancer cells (3) Hybridoma culture supernatant is added to culture medium of human cancer cells In addition, screening for uptake into cells by immunostaining In combination with these screens, hybridoma clones are evaluated to produce monoclonal antibodies with high specificity for mortalin antigen and high uptake capacity into cancer cells. The clones were narrowed down to several types.
  • Method for measuring anti-cancer activity of peptide antibody Tumor growth is observed by forming tumor buds in the flank of nude mice by subcutaneous injection of human fibrosarcoma cells (10 7 ), and monitoring the change in tumor weight by injecting each peptide antibody. The inhibitory activity of was observed.
  • the anti-mortalin monoclonal antibody (CNTL) having anticancer activity for comparison with the peptide antibody of the present invention has a function of internalizing in cancer cells obtained by immunizing mice with the full length of mouse mortalin. It is monoclonal antibody 37-6 (hybridoma 37 strain: FERM-BP10408, see Patent Document 1).
  • antibody fragments having antigen-binding sites such as Fab fragments obtained by enzymatic hydrolysis of monoclonal antibody C-26 or C-69 with papain or the like, F (ab ′) 2 fragments, H chains and L chains
  • Fab fragments obtained by enzymatic hydrolysis of monoclonal antibody C-26 or C-69 with papain or the like F (ab ′) 2 fragments, H chains and L chains
  • scFv single chain antibody
  • cDNA obtained from the hybridoma C-26 or C-69 strain mRNA using reverse transcriptase is incorporated into an appropriate vector, which is introduced into a host, and a recombinant antibody is prepared using gene recombination technology.
  • Antibody fragments may be produced.
  • variable region sequence of monoclonal antibody C-26 or C-69 is obtained from the antibody variable region (V region) cDNA derived from the hybridoma C-26 or C-69 strain by the method described in Patent Document 2 and the like.
  • CDR sequences can be determined, and these sequences can be used to make chimeric and humanized antibodies.
  • the epitope sequence of mortalin recognized by the monoclonal antibody C-26 or C-69 of the present invention can be determined in more detail.
  • amino acid sequences of the epitopes recognized by the anti-mortalin antibodies C-26 and C-69 having strong anticancer activity are “EVILVG (SEQ ID NO: 8)” and “DLFGR (SEQ ID NO: 9)”.
  • the amino acid sequences of the epitopes of anti-mortalin antibodies C-131 and C-177 having a moderate anticancer activity are “EVILVGGMT (SEQ ID NO: 10)” and “DLFGRAP (SEQ ID NO: 11)”. (Figs. 15 and 16).
  • the center position of the epitope is shifted, and in particular, the “EVILVG” of the N-terminal epitope "Or” EVILVGGMT "has shifted significantly. From this result, the “EVILVG” region is not a “continuous epitope (linear epitope)” that recognizes the amino acid primary sequence of mortalin, but a three-dimensional structurally configured epitope of mortalin (“stereostructure epitope” or "Discontinuous epitope”)).
  • the monoclonal antibody that recognizes these two epitope regions is more potent than the monoclonal antibody 37-6 (Patent Document 1) that recognizes the single epitope “LFGRAP” studied in the previous application. Has activity. Therefore, an epitope region containing “EVILVGGMT (SEQ ID NO: 10)” or “EVILVG (SEQ ID NO: 8)” in the sequence, and “DLFGRAP (SEQ ID NO: 11)” or “DLFGR (SEQ ID NO: 9)” in the sequence are included.
  • the epitope region containing is a highly immunogenic epitope alone, but by combining these two epitope regions and using it as an immunogen cocktail, we believe that a monoclonal antibody with even stronger anticancer activity can be produced.
  • a 1: 1 immunogen cocktail of SEQ ID NO: 10 and SEQ ID NO: 11 is 2.
  • a hybridoma producing a human mortalin-specific peptide antibody having a cancer cell intrinsic function is obtained by immunizing a mouse prepared according to the “immunoantigen cocktail method” described in 1.
  • the monoclonal antibody has higher specificity for mortalin antigen, higher uptake ability into cancer cells, and higher tumor cell growth inhibitory activity. Can be obtained.
  • Antibody Pharmaceutical Composition The “anti-mortalin peptide antibody” of the present invention is usually mixed with one or more pharmacologically acceptable carriers and used for the purpose of treating or improving cancer. In that case, you may use together with a well-known anticancer agent.
  • the cancer to be treated by the peptide antibody of the present invention is not particularly limited, and includes a wide range of kidney cancer, lung cancer, colon cancer, brain tumor, uterine cancer, ovarian cancer, stomach cancer, skin cancer, breast cancer, prostate cancer, pancreatic cancer, lymphoma and the like. Can be used.
  • the effective dose is selected, for example, in the range of 0.001 mg to 1000 mg per kg body weight.
  • a dose of 0.01 to 100,000 mg / body can be selected per patient, and the dose can be administered before or after clinical symptoms of the disease occur.
  • pharmacologically acceptable carriers and additives include water, pharmaceutically acceptable organic solvents, collagen, polyvinyl alcohol, sodium alginate, water-soluble dextran, polyethylene glycol, human serum albumin (HSA) , Sugar alcohols and sugars such as mannitol and glucose, and surfactants such as Tween 80.
  • the anticancer agent of the present invention is usually administered by parenteral administration route, for example, injection (subcutaneous injection, intravenous injection, intramuscular injection, intraperitoneal injection, etc.), transdermal, transmucosal, nasal, transpulmonary, etc. Administration is also possible.
  • the anticancer agent of the present invention may be a solution preparation or lyophilized for dissolution and reconstitution before use. As an excipient for lyophilization, sugar alcohols and sugars such as mannitol and glucose can be used.
  • the “anti-mortalin peptide antibody” of the present invention can be used as a known fluorescent reagent, enzyme reagent, radioisotope. It can also be used as a reagent for detecting tumor cells by labeling with an element or the like, or by using in combination with a labeled secondary antibody.
  • Example 1 Production of peptide antibody by immune cocktail method (1-1) Preparation of antigenic peptide
  • Two types of 18 amino acid immunogenic peptides “peptide-1” and “peptide-2” were prepared by chemical synthesis.
  • Peptide-1: KAMQDAEVSKSDIGEVIC” contains the common epitope “KAMQDAEVSKSDIGEVI” of the internalized monoclonal antibody, adds “C (Cys) to the C-terminus, and“ Peptide-2: CQDLFGRAPSKAVNPDEDE ”contains mortalin containing the same“ LFGRAP ”. This is a sequence in which “C (Cys) is added to the N-terminus of the derived amino acid sequence (FIG. 1).
  • peptide-1 and peptide-2 a peptide sequence as an epitope of antibody 37-6 is executed in advance, and antigenicity analysis based on hydrophobicity is performed.
  • the antigenic score of “KAMQDAEVSKSDIGEVI” corresponding to peptide-1 was 0.486, and the antigenic score of “QDLFGRAPSKAVNPDEA” corresponding to peptide-2 was as high as 0.650.
  • C (Cys) is added to the terminal. When these two types of peptides are used as antigens, they are used at a ratio of 1: 1.
  • Example 2 Selection of peptide antibody-producing hybridomas (2-1) Screening of hybridomas by Western blotting Cell lysates of normal human cells (TIG-1 cells) and cancer cells (Hela cells) were prepared, and SDS-PAGE was performed. After that, mortalin was detected by Western blotting using the hybridoma culture supernatant. The results are shown in FIG. 2. In the figure, R indicates the result in the recombinant protein extract, and U indicates the result in the human cancer cell (U2OS) solution. In addition, the degree of reactivity was indicated by the number of +. In the lower left frame of FIG. 2, eight selected clones are shown.
  • Example 3 Measurement of anti-cancer activity of peptide antibody and comparison 8 types of anti-peptide monoclonal antibodies (C-26, C-67, C-69, C-) were obtained from the 8 types of hybridoma clones obtained in Example 2. 131, C-133, C-137, C-152 and C-177 antibodies).
  • Human fibrosarcoma cells (10 7 ) were injected subcutaneously into the left and right flank of each nude mouse. Five days later, it was confirmed that tumor buds were visible, and antibody injection was performed. 100 ⁇ g of antibody was injected intravenously once every two days for a total of 10 times. Tumor progression was monitored daily until day 60.
  • the change in body weight of each individual was measured, and the increase in body weight was taken as the tumor weight (measured by Vernier caliper).
  • 5 to 12 show the tumor weight for each antibody as an average value for each individual.
  • FIG. 13 is a diagram in which these are superimposed.
  • PBS is used as the control (CNTL).
  • Example 4 Analysis of epitope recognized by monoclonal antibodies C-26 and C-69
  • SEQ ID NO: 7 amino acid sequence of the following peptide corresponding to positions 368 to 417 of the mortalin amino acid sequence according to the method of Patent Document 2 above.
  • the signal intensities of monoclonal antibodies C131 and C-177 having moderate anticancer activity and monoclonal antibody C-133 having no epitope in this range are also measured.
  • the control ctrl-mouse
  • the control is mouse IgG, indicating that only the secondary antibody was used.
  • the result shows the presence of two epitope regions as shown in FIG. 16, and can be illustrated as a common region on the peptide amino acid sequence in FIG.
  • the amino acid sequences of the epitopes recognized by anti-mortalin antibodies C-26 and C-69 having strong anticancer activity are “EVILVG” and “DLFGR”, and have an anticancer activity having a moderate anticancer activity.
  • the amino acid sequences of the epitopes of mortalin antibodies C-131 and C-177 are two types, “EVILVGGMT” and “DLFGRAP”.
  • the regions of “EVILVG” and “DLFGR” are recognized in common, and it is understood that the C-26 and C-69 antibodies were the strongest in the recognition strength.
  • sequence of the immunogen cocktail used was the sequence of “KAMQDAEVSKSDIGEVI (C)” and “(C) QDLFGRAPSKAVNPDEA”, the N-terminal epitope “EVILVG” or “EVILVGGMT” was significantly shifted.
  • FERM P-21875 Mouse hybridoma C26 strain (deposited at the National Institute of Advanced Industrial Science and Technology Patent Microbiology Depositary, November 27, 2009)
  • Accession number FERM P-21876 Mouse hybridoma C69 strain (deposited at the National Institute of Advanced Industrial Science and Technology Patent Microbiology Depositary, November 27, 2009)

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Abstract

La présente invention concerne des anticorps anti-peptide de mortaline ayant un effet anticancéreux supérieur aux anticorps anti-mortaline connus, et des hybridomes capables de produire les anticorps ci-dessus. La présente invention concerne en outre des agents anticancéreux utilisant les anticorps ci-dessus. À partir de clones d'hybridome obtenus en utilisant, en tant que cocktail immunogène, deux types de peptides contenant des épitopes « LFGRAP » et « KAMQDAEVSKSDIGEVI » d'un anticorps anti-mortaline capable de s'internaliser dans des cellules cancéreuses, la souche C-26 (FERM P-21875) et la souche C-69 (FERM P-21876) d'hybridome capables de produire des anticorps monoclonaux anti-mortaline, ayant une fonction d'internalisation dans des cellules cancéreuses, une spécificité pour l'antigène mortaline, et un excellent effet d'inhibition de la prolifération de cellules cancéreuses in vivo, sont obtenues. La présente invention concerne en outre des agents anticancéreux comprenant ces anticorps monoclonaux en tant que substance active. Les séquences d'épitope reconnues par ces anticorps monoclonaux sont identifiées comme étant « EVILVG » et « DLFGR ».
PCT/JP2010/072086 2009-12-10 2010-12-09 Anticorps anti-peptide de mortaline anticancéreux WO2011071099A1 (fr)

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EP10836019.9A EP2511371B1 (fr) 2009-12-10 2010-12-09 Anticorps anti-peptide de mortaline anticancéreux
US13/514,755 US8586042B2 (en) 2009-12-10 2010-12-09 Hybridomas producing monoclonal anti-mortalin peptide antibodies
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US11427826B2 (en) 2017-08-11 2022-08-30 City of Hopw RNA aptamers against transferrin receptor (TfR)

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KR20170081962A (ko) * 2016-01-05 2017-07-13 한양대학교 산학협력단 켈로이드 또는 비후성 반흔 예방 또는 치료용 조성물

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2014057490A1 (fr) * 2012-10-09 2014-04-17 Ramot At Tel-Aviv University Ltd. Procédés et trousses utilisant la mortaline soluble dans le sang pour prédire un pronostic de cancer
US11105809B2 (en) 2012-10-09 2021-08-31 Ramot At Tel-Aviv University Ltd. Methods and kits for predicting prognosis of cancer using soluble mortalin in blood
US11427826B2 (en) 2017-08-11 2022-08-30 City of Hopw RNA aptamers against transferrin receptor (TfR)

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US8586042B2 (en) 2013-11-19
EP2511371B1 (fr) 2015-02-25
EP2511371A1 (fr) 2012-10-17
US20120302729A1 (en) 2012-11-29
EP2511371A4 (fr) 2013-05-15
JP5641486B2 (ja) 2014-12-17
JPWO2011071099A1 (ja) 2013-04-22

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