WO2021086159A1 - 면역 관문 분자와 결합 가능한 분자가 융합된 단백질 및 이의 용도 - Google Patents
면역 관문 분자와 결합 가능한 분자가 융합된 단백질 및 이의 용도 Download PDFInfo
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Definitions
- the present invention relates to a protein in which a molecule capable of binding to an immune checkpoint molecule is fused and a use thereof.
- cancer is very difficult and complex treatment is required, unlike other disease treatment.
- the methods used for cancer treatment include surgery, radiation therapy, and chemotherapy. If the cancer does not metastasize to other areas and develops locally, cancer can be treated through cancer removal surgery. However, since cancer metastasis occurs in more than 70% of cancer patients, adjuvant therapy must be combined.
- auxiliary treatment regimens radiation therapy that kills cancer cells using high energy radiation is performed, and the radiation therapy inhibits the proliferation of cancer cells when irradiating the cancer cells with radiation, so that new cancer cells cannot be generated. Prevents further division.
- this method has a problem that there is a side effect of affecting not only cancer cells but also normal cells.
- Chemotherapy is an adjuvant therapy in which a drug is used to kill cancer cells after surgery, and is performed for the purpose of killing invisible cancer cells.
- the chemotherapy has a problem that side effects such as vomiting, diarrhea, and hair loss follow.
- Immunotherapy methods have recently emerged to minimize these side effects.
- Immunotherapy is a method of treating cancer using the patient's immune response, and can even prevent cancer.
- Cancer immunotherapy is a treatment method that activates cancer-specific immune cells by administering an antigen that causes tumor formation, as in the principle of a vaccine, and then causes the activated immune cells to specifically attack the cancer in the body.
- the inactivated immune cells are activated as cancer-specific memory immune cells, so that when cancer occurs, cancer cells can be specifically attacked.
- TAA tumor-associated antigen
- TSA Tumor-specific antigen
- TSA tumor-specific antigen
- neo-antigens which are found in various tumor types such as lung cancer and kidney cancer, but mainly found in melanoma, are antigens that are newly generated by potential gene activity of individuals with cancer or mutations in the DNA part. This antigen is very important in producing a'customized cancer vaccine' based on the patient's individual genetic information.
- Non-Patent Document 1 Polymers are widely used as carriers of these cancer-specific antigens in the body, and when a cancer antigen is immobilized on the surface of the polymer for transport of the cancer-specific antigen in the body, the cancer-specific antigen must be exposed to the particle surface through chemical binding. .
- Cancer immunotherapy uses the patient's immune system compared to conventional anti-cancer treatment methods, so the side effects are low, the therapeutic effect can be sustained for a long time due to the formation of immune memory, and the effect on general cells is low due to the principle of tumor antigen-specific recognition. It has the advantage of having few side effects.
- cancer immunotherapy has been receiving explosive attention enough to be selected by Science as Breakthrough of the year 2013.
- immune checkpoint PD-1 Programmed cell death protein 1
- PD-L1 PD1 ligand
- Cancer immunotherapy is likely to be used in clinical settings as a standard treatment for cancer.
- immune checkpoints PD1/PD-L1 or CTLA4-specific immune activity blocking antibodies cannot fundamentally enhance the activity of immune cells, and are self-generated due to the long residence time in the body and the Fc domain of the antibody. Side effects such as immune diseases occur, and there is also a disadvantage in terms of economics due to the high cost of antibody treatment due to the high consumption amount as well as the high production cost due to antibody production and purification of the animal cell system.
- An object of the present invention is to provide a novel protein capable of binding to an immune checkpoint molecule.
- An object of the present invention is to provide a novel protein capable of killing cancer cells by preventing immune checkpoint molecules from inactivating T cells.
- An object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of cancer comprising the above novel protein.
- An object of the present invention is to provide a method for treating cancer comprising the above novel protein.
- the present invention provides a ferritin protein in which a molecule capable of binding to an immune checkpoint molecule is fused to an outer surface.
- the protein of the present invention may be a spherical protein formed by self-assembly of 24 ferritin monomers in which a molecule capable of binding to the immune checkpoint molecule is fused.
- the molecule capable of binding to the immune checkpoint molecule may be fused to at least one of adjacent ⁇ -helixes of the ferritin monomer.
- the molecule capable of binding to the immune checkpoint molecule may be fused to the N-terminus or C-terminus of the ferritin monomer.
- the molecule capable of binding to the immune checkpoint molecule may be fused to the A-B loop, B-C loop, C-D loop, or D-E loop of the ferritin monomer.
- a molecule capable of binding to the immune checkpoint molecule may be fused between the N-terminus and A helix of the ferritin monomer or between the E helix and C-terminus.
- the protein of the present invention may be mutated so as to decrease the binding ability to the human transferrin receptor.
- At least one of the ferritin monomers constituting it may be one in which amino acids 14, 15, 22, 81 or 83 in the sequence of SEQ ID NO: 1 are substituted with alanine, glycine, valine, or leucine.
- the ferritin protein of the present invention may have an avidity (K) for a transferrin receptor satisfying the following Equation 1:
- K [P][T]/[PT]
- [P] represents the concentration of ferritin protein in equilibrium of the binding reaction between the ferritin protein and the transferrin receptor
- [T] is the equilibrium.
- concentration of the transferrin receptor in the state is indicated
- [PT] indicates the concentration of the complex of the ferritin protein and the transferrin receptor in the equilibrium state).
- the binding force to the transferrin receptor may be the binding force to the human transferrin receptor.
- the immune checkpoint molecule is Her-2/neu, VISTA, 4-1BBL, Galectin-9, Adenosine A2a receptor, CD80, CD86, ICOS, ICOSL, BTLA, OX-40L, CD155, BCL2, MYC, PP2A, BRD1, BRD2, BRD3, BRD4, BRDT, CBP, E2F1, MDM2, MDMX, PPP2CA, PPM1D, STAT3, IDH1, PD1, CTLA4, PD-L1, PD-L2, LAG3, TIM3, TIGIT, BTLA, SLAMF7, 4- 1BB, OX-40, ICOS, GITR, ICAM-1, BAFFR, HVEM, LFA-1, LIGHT, NKG2C, SLAMF7, NKp80, LAIR1, 2B4, CD2, CD3, CD16, CD20, CD27, CD28, CD40L, CD48, It may be any one selected from the group consisting of CD52,
- the molecule capable of binding to the immune checkpoint molecule may be a ligand, an antibody, or a fragment thereof for the immune checkpoint molecule.
- ferritin may be a human ferritin heavy chain.
- the ferritin protein of the present invention may be present in a water-soluble fraction of 40% or more in the E. coli production system.
- the present invention provides a pharmaceutical composition for the treatment or prevention of cancer comprising the ferritin protein of the present invention.
- the pharmaceutical composition of the present invention is brain cancer, head and neck cancer, bladder cancer, breast cancer, cervical cancer, colon cancer, colorectal cancer, endometrial cancer, esophageal cancer, leukemia, lung cancer, liver cancer, ovarian cancer, pancreatic cancer, prostate cancer, rectal cancer, kidney cancer, stomach cancer, Testicular cancer, uterine cancer, vascular tumor, squamous cell carcinoma, adenocarcinoma, small cell carcinoma, melanoma, glioma, neuroblastoma, sarcoma, laryngeal cancer, parotid carcinoma, biliary tract cancer, thyroid cancer, actinic keratosis, acute lymphocytic leukemia, acute myeloid leukemia, adenocarcinoma Carcinoma, adenoma, glandular squamous cell carcinoma, anal duct cancer, anal cancer, anal rectal cancer, astrocytoma, large vaginal gland carcinoma, basal cell
- the pharmaceutical composition of the present invention may further include a protein comprising a ferritin monomer fused to a disease antigen epitope is self-assembled, and a binding force (K) to a transferrin receptor satisfies the following equation (4):
- K [P][T]/[PT]
- [P] represents the concentration of the protein in equilibrium state of the binding reaction between the protein and the transferrin receptor
- [T] is the above
- the concentration of the transferrin receptor in the equilibrium state is indicated
- [PT] indicates the concentration of the complex of the protein and the transferrin receptor in the equilibrium state).
- Disease antigen epitopes in the present invention are gp100, MART-1, Melna-A, MAGE-A3, MAGE-C2, Mammaglobin-A, proteinsase-3, mucin-1, HPV E6, LMP2, PSMA, GD2, hTERT, PAP, ERG, NA17, ALK, GM3, EPhA2, NA17-A, TRP-1, TRP-2, NY-ESO-1, CEA, CA 125, AFP, Survivin, AH1, ras, G17DT, MUC1, Her-2/neu , E75, p53, PSA, HCG, PRAME, WT1, URLC10, VEGFR1, VEGFR2, E7, Tyrosinase peptide, B16F10, EL4 or neoantigen (neoantigen).
- the pharmaceutical composition of the present invention may be for administration of the two kinds of proteins in combination.
- the proteins of the species may be administered simultaneously, individually or sequentially.
- 1A shows a schematic diagram of an expression vector for producing the protein of the present invention in which a tumor antigen is expressed
- B shows the structure of the produced protein.
- FIG. 2 is a schematic diagram showing the binding site of the tumor antigen and transferrin receptor (TfR) on the surface of the gp100-huHF nanoparticles prepared according to the present invention.
- TfR tumor antigen and transferrin receptor
- FIG. 3 shows the TEM image and DLS results of the gp100-huHF protein of the present invention.
- Figure 4 is a result of measuring the binding ability of the gp100-huHF protein of the present invention and the transferrin receptor (TfR).
- FIG. 5 is a schematic diagram of an expression vector for preparing an immune checkpoint inhibitor (huHF-PD1 protein) into which a PD1 domain capable of binding to PD-L1 is inserted; structure of the gp100-huHF protein; TEM image of the gp100-huHF protein of the present invention; Diameter distribution diagram of the gp100-huHF protein of the present invention; And huHF-PD1 protein and PD1 ligand (PD-L1), huHF-TPP1 (AB loop, CD loop), and ⁇ PD-L1 HCDR3 (CD loop, C-terminal).
- huHF-PD1 protein an immune checkpoint inhibitor
- Figure 6 shows the results of cellular uptake by dendritic cells of the protein of the present invention.
- 7A is a result of comparing the targeting efficiency of huHF protein and huHF-PD1 protein to cancer cells CT-26 and B16F10 through fluorescence images;
- 7B is a comparison of the targeting efficiency of huHF protein and huHF- ⁇ PD-L1 HCDR3 (CD loop, C-terminal) to CT-26 cells through fluorescence images;
- 7C is a comparison of the targeting efficiency of huHF protein, huHF-TPP1, and huHF-smPD1 to CT-26 cells through fluorescence images.
- 11A is a result of confirming whether the OVA-huHF protein can increase OVA peptide antigen presentation of antigen-presenting cells through flow cytometry (FACS), and B is a result of confirming the expression level of DC maturation marker of the protein.
- FACS flow cytometry
- the results of MHC-II, CD80, CD40, and CD86 are from the left in the bar graph of each group of B.
- FIG. 12 shows a schematic diagram and experimental results of an experimental method for confirming the ability of gp100-huHF protein to inhibit tumor antigens.
- FIG. 13 shows a schematic diagram and experimental results of an experimental method for confirming the tumor formation inhibitory effect of huHF-PD1 protein in CT26 (colorectal cancer cells) and B16F10 (melanoma cells) in an animal model.
- CT26 colonal cancer cells
- B16F10 melanoma cells
- 15A is a comparison of the T-cell mediated apoptosis efficiency of PD-L1 antibody and huHF-PD1 protein in cancer cells CT26 and B16F10;
- B is a comparison of the T-cell activity response of PD-L1 antibody and huHF-PD1 protein in cancer cells CT26 and B16F10;
- C shows T-cell activating responses to tumor antigens, respectively, by combination treatment of AH1-huHF protein, gp100-huHF protein, and huHF-PD1.
- CT26 colonrectal cancer cells
- AH1-huHF protein and/or huHF-PD1 protein show the results of suppression of tumor recurrence in CT26 (colorectal cancer cells) according to treatment with AH1-huHF protein and/or huHF-PD1 protein.
- the results are PBS, AH1-huHF, ⁇ -PD-L1, PD1-huHF, AH1-huHF + ⁇ -PD-L1, AH1-huHF + PD1-huHF.
- Figure 19 is NA-gp100-huHF
- Figure 20 is EC-gp100-huHF
- Figure 21 is D in -gp100-huHF
- Figure 22 is Ein0gp100-huHF
- Figure 23 is a vector schematic diagram for each preparation of msmPD1-huHF and its It confirms the production of the protein.
- 25 is a schedule for evaluating the tumor inhibitory ability of the huHF-PD-L1-TIGIT dual blocker.
- 26 and 27 are the results of evaluating the tumor suppression ability of the huHF-PD-L1-TIGIT dual blocker.
- FIG. 30 is a schematic diagram of a vector of huHF- ⁇ PD-L1 HCDR3, and the production and self-assembly of the protein were confirmed.
- Fig. 31 is a schematic diagram of a vector of huHF- ⁇ PD1 HCDR3, and the production and self-assembly of the protein are confirmed.
- FIG. 32 is a schematic diagram of a vector of huHF- ⁇ CTLA4 HCDR3, and production and self-assembly of the protein were confirmed.
- Fig. 33 is a schematic diagram of a vector of huHF- ⁇ TIGIT HCDR3, and production and self-assembly of the protein are confirmed.
- Fig. 34 is a schematic diagram of a vector of huHF- ⁇ LAG3 HCDR3, and the production and self-assembly of the protein are confirmed.
- Fig. 35 is a schematic diagram of a vector of huHF- ⁇ TIM3 HCDR3, and production and self-assembly of the protein are confirmed.
- FIG. 36 is a schematic diagram of a vector of huHF- ⁇ PD-L1- ⁇ TIGIT, and production and self-assembly of the protein were confirmed.
- the present invention relates to a ferritin protein (protein A) in which a molecule capable of binding to an immune checkpoint molecule is fused to an outer surface.
- protein A ferritin protein
- Ferritin may be ferritin derived from humans, animals and microorganisms.
- Human ferritin is composed of a heavy chain (21 kDa) and a light chain (19 kDa), and exhibits the property of forming spherical nanoparticles through the self-assembly ability of the monomers constituting the ferritin.
- Ferritin can form a self-assembly having a spherical three-dimensional structure by gathering 24 monomers.
- the outer diameter is about 12 nm and the inner diameter is about 8 nm.
- the structure of the ferritin monomer is a form in which five ⁇ -helix structures, namely A helix, B helix, C helix, D helix, and E helix are sequentially linked, and each ⁇ -helix structure, called a loop, is formed. It includes a linking atypical polypeptide moiety.
- the loop is a region that is not structurally damaged even if a peptide or a small protein antigen is inserted into ferritin.
- a peptide-ferritin fusion protein monomer in which a peptide such as an epitope is located on a monomer of ferritin can be prepared.
- the loop connecting the A helix and the B helix is the AB loop
- the loop connecting the B helix and the C helix is the BC loop
- the loop connecting the C helix and the D helix is the CD loop
- the loop connecting the D helix and the E helix is the DE It is called a loop.
- Ferritin may be a ferritin heavy chain, specifically, a human ferritin heavy chain.
- the human ferritin heavy chain may be a protein represented by the amino acid sequence of SEQ ID NO: 1 derived from human, and in the present specification, the ferritin may be used interchangeably with'human ferritin heavy chain' or'huHF'.
- ferritin In ferritin, several of its monomers are self-assembled to form an organized structure or pattern.
- the ferritin protein of the present invention is a nano-scale particle.
- 24 ferritin monomers in which a molecule capable of binding to an immune checkpoint molecule is fused can be self-assembled to form a ferritin protein.
- the ferritin protein (protein A) of the present invention may be spherical.
- the particle diameter may be 8 to 50 nm. More specifically, it may be 8nm to 50nm, 8nm to 45nm, 8nm to 40nm, 8nm to 35nm, 8nm to 30nm, 8nm to 25nm, 8nm to 20nm, 8nm to 15nm.
- the ferritin protein (protein A) of the present invention is a fusion of a molecule capable of binding to an immune checkpoint molecule on an outer surface. As long as the molecule capable of binding to the immune checkpoint molecule is fused to the outer surface of the ferritin protein, it is not necessary that the molecule capable of binding to the immune checkpoint molecule is fused to all ferritin monomers constituting the ferritin protein.
- a molecule capable of binding to an immune checkpoint molecule is fused to self-assemble to form a ferritin protein (protein A)
- a molecule capable of binding to an immune checkpoint molecule is fused to at least one ferritin monomer out of 24 ferritin monomers. All you need In order to increase the density of molecules that can bind to the immune checkpoint molecule on the outer surface of the ferritin protein, all 24 ferritin monomers can be fused with the immune checkpoint molecule and bindable molecules.
- Each ferritin monomer constituting the ferritin protein may be fused with one or more molecules capable of binding to an immune checkpoint molecule.
- Each ferritin monomer constituting the ferritin protein may be fused with a molecule capable of binding to a different immune checkpoint molecule.
- Each ferritin monomer constituting the ferritin protein may be a fusion of different molecules capable of binding to the same immune checkpoint molecule.
- the immune checkpoint molecule binds to T cells and inactivates T cells.
- These immune checkpoint molecules are, for example, Her-2/neu, VISTA, 4-1BBL, Galectin-9, Adenosine A2a receptor, CD80, CD86, ICOS, ICOSL, BTLA, OX-40L, CD155, BCL2, MYC, PP2A, BRD1, BRD2, BRD3, BRD4, BRDT, CBP, E2F1, MDM2, MDMX, PPP2CA, PPM1D, STAT3, IDH1, PD1, CTLA4, PD-L1, PD-L2, LAG3, TIM3, TIGIT, BTLA, SLAMF7, 4- 1BB, OX-40, ICOS, GITR, ICAM-1, BAFFR, HVEM, LFA-1, LIGHT, NKG2C, SLAMF7, NKp80, LAIR1, 2B4, CD2, CD3, CD16, CD20, CD27, CD28, CD40L
- Molecules capable of binding the immune checkpoint molecule may be a ligand for the immune checkpoint molecule, an antibody, or a fragment thereof.
- Molecules capable of binding to the immune checkpoint molecule can be fused to any ferritin monomer as long as it can be exposed to the outer surface of the ferritin protein. Molecules capable of binding to the immune checkpoint molecule are fused to a site that does not interfere with the self-assembly of the ferritin monomer.
- Molecules that can bind to the immune checkpoint molecule are fused inside the ferritin monomer, which can change the structure of the ferritin protein.
- the internally embedded part can protrude to the outside by the fusion of the immune checkpoint molecule and the binding molecule.
- the part protruding to the outside among the constituent parts of the ferritin monomer is the immune checkpoint molecule. It can be incorporated into the inside by fusion of a molecule capable of binding with.
- the molecule capable of binding to the immune checkpoint molecule is fused to a position where the ferritin protein (protein A) decreases the binding ability with the human transferrin receptor or interferes with the binding ability with the human transferrin receptor.
- ferritin protein protein A
- Molecules capable of binding to an immune checkpoint molecule are not limited to a specific range of structures, molecular weights, and amino acid lengths.
- a molecule capable of binding an immune checkpoint molecule may be, for example, a ligand, antibody, or fragment thereof whose amino acid length is 25aa or less. More specifically, amino acids whose length is 25aa or less, 24aa or less, 23aa or less, 22aa or less, 21aa or less, 20aa or less, 19aa or less, 18aa or less, 17aa or less, 16aa or less, 15aa or less, 14aa or less, 13aa or less, 12aa or less, 11aa Hereinafter, it may be 10aa or less, 9aa or less, 8aa or less, 7aa or less, 6aa or less, 5aa or less.
- the length of the amino acid may be 3aa or more, 4aa or more, 5aa or more, 6aa or more, 7aa or more, 8aa or more, 9aa or more, 10aa or more.
- the fusion site of the molecule capable of binding to the immune checkpoint molecule is not limited to a specific position, such as between adjacent ⁇ -helixes of ferritin monomers, N-terminus, C-terminus, AB loop, BC loop, CD loop, DE loop, N -It can be fused between the end and A helix, between the E helix and C-end, inside the helix, etc.
- ferritin protein (protein A) of the present invention is designed to bind to an immune checkpoint molecule, it is preferable that it does not bind well to the transferrin receptor.
- the ferritin protein of the present invention satisfies the following equation 1 for the transferrin receptor:
- K [P][T]/[PT]
- [P] represents the concentration of ferritin protein in equilibrium of the binding reaction between the ferritin protein and the transferrin receptor
- [T] is the equilibrium.
- concentration of the transferrin receptor in the state is indicated
- [PT] indicates the concentration of the complex of the ferritin protein and the transferrin receptor in the equilibrium state).
- the avidity may be, for example, a binding force to a human transferrin receptor.
- the K value in Equation 1 is 100nM or more, 110nM or more, 120nM or more, 125nM or more, 125nM or more, 150nM or more, 200nM or more, 210nM or more, 220nM or more, 230nM or more, 240nM or more, 250nM or more, 260nM or more, 270nM or more, 280nM Or more, 290nM or more, 300nM or more, 350nM or more, 400nM or more, 450nM or more, 500nM or more, 550nM or more, 600nM or more, 700nM or more, 800nM or more, 900nM or more, 1000nM or more.
- the avidity (K) to the transferrin receptor is measured in an equilibrium state of the binding reaction between the ferritin protein (A) of the present invention and the transferrin receptor.
- concentration of ferritin protein ([P]), the concentration of the transferrin receptor ([T]), and the concentration of the complex of the protein of the present invention and the transferrin receptor ([PT]) in an equilibrium state can be measured by various known methods. .
- the binding force (K) to the transferrin receptor can be measured according to, for example, a Microscale Thermophoresis (MST) method.
- MST Microscale Thermophoresis
- Monolith NT.115 is an MST measuring device.
- Equation 1 The concentration of Equation 1 may be obtained by utilizing the following Equations 2 and 3.
- [PT] 1/2 x (([P 0 ]+[A 0 ]+[P][T]/[PT])-(([P 0 ]+[T 0 ]+ ([P][T ]/[PT]) 2 )-4 x [P 0 ] x [T 0 ]) 1/2 )
- [PT] is the concentration in the parallel state of reaction of the complex of ferritin protein and transferrin receptor
- P 0 is the initial concentration of ferritin protein
- T 0 is the initial concentration of transferrin receptor
- [P] is the reaction parallel of ferritin protein.
- the concentration in the state, [T] represents the concentration in the parallel state of the reaction of the transferrin receptor, respectively).
- [PT] is the concentration in the reaction parallel state of the complex of the ferritin protein and the transferrin receptor
- P 0 is the initial concentration of the ferritin protein
- X is the ratio of the protein complexed with the transferrin receptor in the ferritin protein
- the ferritin protein (protein A) of the present invention can fuse an immune checkpoint molecule and a molecule capable of binding to a site involved in binding to the transferrin receptor in order to lower the binding ability to the transferrin receptor.
- the molecule may be fused so that a molecule capable of binding to an immune checkpoint molecule is located in the B-C loop and A helix portions of the ferritin protein of the present invention.
- the ferritin protein (protein A) of the present invention may be produced in a microorganism expressing a sequence encoding the protein.
- microorganisms known in the art may be used without limitation.
- it may be E. coli, specifically BL21 (DE3), but is not limited thereto.
- the obtained protein In the case of producing a protein by a microbial system, the obtained protein must be present in a dissolved state in the cytoplasm to facilitate separation/purification. In many cases, the produced protein exists in an aggregated state as an inclusion body.
- the ferritin protein of the present invention has a high percentage dissolved in the cytoplasm in the microbial production system. It is easy to separate/purify and use.
- the ferritin protein (protein A) of the present invention can be prepared, for example, in a state in which the water-soluble fraction ratio of the total protein is 40% or more in the E. coli system for producing the same. Specifically, it may be 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, 95% or more.
- the upper limit may be, for example, 100%, 99%, 98%, 97%, 96%, and the like.
- the present invention provides a pharmaceutical composition for preventing or treating cancer comprising the above ferritin protein (protein A). All of the above descriptions of the ferritin protein apply as it is to the ferritin protein as an active ingredient of the pharmaceutical composition of the present application.
- the pharmaceutical composition of the present invention may contain a pharmaceutically acceptable carrier.
- pharmaceutically acceptable carrier refers to a carrier or diluent that does not significantly irritate an organism and does not impair the biological activity and properties of an administered component.
- the pharmaceutically acceptable carrier in the present invention may be used by mixing one component or one or more of these components, including saline, sterile water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol, and If necessary, other conventional additives such as antioxidants, buffers and bacteriostatic agents may be added, and formulated in the form of an injection suitable for injection into tissues or organs.
- a target organ-specific antibody or other ligand may be used in combination with the carrier so that it can specifically act on the target organ.
- composition of the present invention may further include a filler, an excipient, a disintegrant, a binder or a lubricant.
- compositions of the present invention may be formulated using methods known in the art to provide rapid, sustained or delayed release of the active ingredient after administration to a mammal.
- the pharmaceutical composition may be an injection formulation, and may be administered intravenously, but is not limited thereto.
- the term "effective amount” means an amount necessary to delay the onset or progression of a specific disease to be treated or to entirely enhance it.
- the composition may be administered in a pharmaceutically effective amount. It is obvious to a person skilled in the art that the appropriate total daily use amount of the pharmaceutical composition can be determined by the treating physician within the range of correct medical judgment.
- a specific pharmaceutically effective amount for a specific patient is a specific composition, including the type and extent of the reaction to be achieved, whether or not other agents are used in some cases, the patient's age, weight, general health status, and sex. And it is preferable to apply differently according to various factors including diet, administration time, administration route and secretion rate of the composition, treatment period, drugs used with or concurrently with the specific composition, and similar factors well known in the medical field.
- the pharmaceutical composition may be accompanied by an instruction in connection with the packaging in a form directed by a government agency in charge of the manufacture, use and sale of drugs, if necessary, and the instruction may be in the form of a composition or a human or It represents private interest approval for administration to animals, and may be, for example, a label approved by the US Food and Drug Administration for the prescription of drugs.
- Cancers include, for example, brain cancer, head and neck cancer, bladder cancer, breast cancer, cervical cancer, colon cancer, colorectal cancer, endometrial cancer, esophageal cancer, leukemia, lung cancer, liver cancer, ovarian cancer, pancreatic cancer, prostate cancer, rectal cancer, kidney cancer, stomach cancer, testicular cancer.
- Uterine cancer vascular tumor, squamous cell carcinoma, adenocarcinoma, small cell carcinoma, melanoma, glioma, neuroblastoma, sarcoma, laryngeal cancer, parotid adenocarcinoma, biliary tract cancer, thyroid cancer, actinic keratosis, acute lymphocytic leukemia, acute myeloid leukemia, adenocystic carcinoma , Adenoma, glandular squamous cell carcinoma, anal duct cancer, anal cancer, anal rectal cancer, astrocytoma, large vaginal gland cancer, basal cell carcinoma, biliary cancer, bone cancer, bone marrow cancer, bronchial cancer, bronchial carcinoma, carcinoid, cholangiocarcinoma, chronic lymph Constituent leukemia, chronic myelogenous leukemia, clear cell carcinoma, connective tissue cancer, cyst adenoma, digestive system cancer, du
- the present invention is made by self-assembly of a ferritin monomer to which a disease antigen epitope is fused, and a pharmaceutical for the treatment or prevention of cancer further comprising a protein (protein B) having a binding force (K) to the transferrin receptor satisfies the following equation (4)
- a protein protein B having a binding force (K) to the transferrin receptor satisfies the following equation (4)
- the composition is provided:
- K [P][T]/[PT]
- [P] represents the concentration of the protein in equilibrium state of the binding reaction between the protein and the transferrin receptor
- [T] is the above
- the concentration of the transferrin receptor in the equilibrium state is indicated
- [PT] indicates the concentration of the complex of the protein and the transferrin receptor in the equilibrium state).
- the binding force to the transferrin receptor may be the binding force to the human transferrin receptor.
- the coupling force represented by Equation 4 may be obtained by the same method as the method for obtaining the coupling force represented by Equation 1, but is not limited thereto.
- the ferritin protein (protein B) to which the disease antigen epitope of the present invention is fused has an avidity (K) of 700 nM or less, 600 nM or less, 500 nM or less, 400 nM or less, 300 nM or less, 200 nM or less, 150 nM or less, 125 nM or less, 100 nM or less.
- K avidity
- it may be 50nM or less, 40nM or less, 30nM or less, 20nM or less, 10nM or less.
- the lower limit may be 0.5nM, 1nM, 1.5nM, 2nM, 2.5nM, 3nM, 3.5nM, 4nM, 4.5nM, 5nM, but is not limited thereto.
- Disease antigens can be antigens of any disease that can be prevented, treated, alleviated or ameliorated by an immune response.
- the disease antigen may be a cell surface antigen of a cancer cell, a pathogen cell, or a cell infected with a pathogen.
- the specific site that determines the antigen specificity of a disease antigen is a disease antigen epitope.
- the disease is, for example, cancer or an infectious disease.
- Cancers include, for example, brain cancer, head and neck cancer, bladder cancer, breast cancer, cervical cancer, colon cancer, colorectal cancer, endometrial cancer, esophageal cancer, leukemia, lung cancer, liver cancer, ovarian cancer, pancreatic cancer, prostate cancer, rectal cancer, kidney cancer, stomach cancer, testicular cancer, uterine cancer.
- Vascular tumor squamous cell carcinoma, adenocarcinoma, small cell carcinoma, melanoma, glioma, neuroblastoma, sarcoma, laryngeal cancer, parotid adenocarcinoma, biliary tract cancer, thyroid cancer, actinic keratosis, acute lymphocytic leukemia, acute myeloid leukemia, adenocyst carcinoma, adenoma , Glandular squamous cell carcinoma, anal duct cancer, anal cancer, anal rectal cancer, astrocytoma, large vaginal esophagus cancer, basal cell carcinoma, biliary cancer, bone cancer, bone marrow cancer, bronchial cancer, bronchial carcinoma, carcinoid, cholangiocarcinoma, chronic lymphocytic leukemia , Chronic myeloid leukemia, clear cell carcinoma, connective tissue cancer, cyst adenoma, digestive system
- the infectious disease can be, for example, a viral, bacterial, fungal, parasitic or prion infection.
- Cancer antigen epitopes are gp100, MART-1, Melna-A, MAGE-A3, MAGE-C2, Mammaglobin-A, proteinsase-3, mucin-1, HPV E6, LMP2, PSMA, GD2, hTERT, PAP, ERG, NA17.
- ALK ALK
- CEA CA 125, AFP, Survivin, AH1, ras, G17DT, MUC1, Her-2/neu, E75, p53, PSA, HCG, PRAME, WT1, URLC10, VEGFR1, VEGFR2, E7, Tyrosinase peptide, B16F10, EL4 or neoantigen.
- Neoantigen refers to an immunogenic peptide that is induced and formed by somatic mutations in tumor cells. Neoantigens form complexes with MHC I and migrate to the surface of tumor cells and can be displayed as antigen epitopes. T-cell receptors (TCRs) recognize the neoantigen-MHCI complex to trigger an immune response. To induce.
- the disease antigen epitope is not limited to a specific length as long as it can be fused to the ferritin monomer.
- the disease antigen epitope is not limited to a specific length as long as it does not interfere with self-assembly of the ferritin monomer.
- the disease antigen epitope can be fused to any of the ferritin monomers.
- the disease antigen epitope is fused to a site that does not interfere with self-assembly of the ferritin monomer.
- the disease antigen epitope is preferably fused to the ferritin monomer so that it is exposed to the protein surface for binding to the human transferrin receptor.
- Disease antigen epitopes are, for example, whose amino acid length is 25aa or less, 24aa or less, 23aa or less, 22aa or less, 21aa or less, 20aa or less, 19aa or less, 18aa or less, 17aa or less, 16aa or less, 15aa or less, 14aa or less, 13aa or less, 12aa
- it may be 11aa or less, 10aa or less, 9aa or less, 8aa or less, 7aa or less, 6aa or less, 5aa or less.
- the disease antigen epitope may be, for example, the amino acid length of 3aa or more, 4aa or more, 5aa or more, 6aa or more, 7aa or more, 8aa or more, 9aa or more, 10aa or more.
- the fusion of the disease antigen epitope to the ferritin monomer may improve the binding ability of the self-assembled protein (protein B) of the ferritin monomer to the human transferrin receptor.
- protein B self-assembled protein
- a portion incorporated into the inside may protrude outward after binding of the disease antigen epitope.
- the fusion site of the disease antigen epitope in the perintin monomer is not limited to a specific position, such as between adjacent ⁇ -helixes, N-terminus, C-terminus, AB loop, BC loop, CD loop, DE loop, N-terminus It can be fused between the A and A helix, the E helix and the C-terminal, and the inside of the helix.
- the disease antigen epitope can be fused to at least one of adjacent ⁇ -helixes.
- the disease antigen epitope can be fused to the N-terminus or C-terminus of the ferritin monomer.
- the disease antigen epitope may be fused to the A-B loop, B-C loop, C-D loop or D-E loop of the ferritin monomer.
- the disease antigen epitope may be fused between the N-terminus and A helix of the ferritin monomer or between the E helix and C-terminus.
- the disease antigen epitope may be fused to the interior of at least one of each helix of the ferritin monomer.
- the protein (protein B) of the present invention is a self-assembled ferritin monomer to which a disease antigen epitope is fused.
- Ferritin is a self-assembled protein that forms an aggregate by forming an organizational structure or pattern on its own when several monomers are collected, and it is possible to form nanoscale proteins without additional manipulation.
- ferritin monomer to which the disease antigen epitope according to the present invention is fused also forms a self-assembled protein.
- 24 ferriline monomers can be self-assembled to form spherical particles.
- the particle diameter may be, for example, 8 to 50 nm. Specifically, it may be 8nm to 50nm, 8nm to 45nm, 8nm to 40nm, 8nm to 35nmm, 8nm to 30nm, 8nm to 25nm, 8nm to 20nm, 8nm to 15nm, etc., but is not limited thereto.
- the ferritin monomer to which the disease antigen epitope is fused is self-assembled, and the protein that binds to the transferrin receptor (protein B) binds to the transferrin receptor (TfR) present on the surface of dendritic cells, which are antigen-presenting cells. Accordingly, the fused antigen epitope is introduced and presented to the dendritic cells, and the immune system recognizes the antigen so that the immune response can be performed.
- the protein (protein B) that is formed by self-assembly of the ferritin monomer to which the disease antigen epitope is fused and binds to the transferrin receptor may be a linker peptide further included between the human ferritin heavy chain protein and the disease antigen epitope.
- the linker peptide is not limited as long as it is a sequence for enhancing the surface expression of a protein by imparting flexibility to the epitope, but may have an amino acid sequence of SEQ ID NO: 36 to SEQ ID NO: 38, for example.
- the linker peptide may have a length capable of securing an appropriate space between disease antigen epitopes.
- the linker peptide may be a peptide consisting of 1 to 20, 3 to 18, 4 to 15, and 8 to 12 amino acids. By adjusting the length and/or amino acid composition of the linker peptide, the spacing and orientation between disease antigen epitopes can be controlled.
- the pharmaceutical composition of the present invention may be administered in combination with the two proteins (proteins A and B).
- the order of administration at the time of combined administration is not limited, and for example, two types of proteins may be administered simultaneously, or may be administered individually or sequentially.
- sequential administration a ferritin protein in which a molecule capable of binding to an immune checkpoint molecule is fused to an outer surface may be administered first, or a protein formed by self-assembly of a ferritin monomer to which a disease antigen epitope is fused may be administered first.
- the present invention provides a method for treating cancer comprising the above ferritin protein (protein A). All of the matters described above with respect to the ferritin protein apply as it is to the ferritin protein as an active ingredient in the cancer treatment method of the present application.
- the treatment method of the present invention includes the step of administering the ferritin protein (protein A) of the present application to a subject suffering from cancer.
- the individual suffering from cancer may be an animal suffering from cancer, specifically a mammal suffering from cancer, and more specifically may be a human suffering from cancer.
- Cancers include, for example, brain cancer, head and neck cancer, bladder cancer, breast cancer, cervical cancer, colon cancer, colorectal cancer, endometrial cancer, esophageal cancer, leukemia, lung cancer, liver cancer, ovarian cancer, pancreatic cancer, prostate cancer, rectal cancer, kidney cancer, stomach cancer, testicular cancer.
- Uterine cancer vascular tumor, squamous cell carcinoma, adenocarcinoma, small cell carcinoma, melanoma, glioma, neuroblastoma, sarcoma, laryngeal cancer, parotid adenocarcinoma, biliary tract cancer, thyroid cancer, actinic keratosis, acute lymphocytic leukemia, acute myeloid leukemia, adenocystic carcinoma , Adenoma, glandular squamous cell carcinoma, anal duct cancer, anal cancer, anal rectal cancer, astrocytoma, large vaginal gland cancer, basal cell carcinoma, biliary cancer, bone cancer, bone marrow cancer, bronchial cancer, bronchial carcinoma, carcinoid, cholangiocarcinoma, chronic lymph Constituent leukemia, chronic myelogenous leukemia, clear cell carcinoma, connective tissue cancer, cyst adenoma, digestive system cancer, du
- Protein (Protein A) can be administered in a therapeutically effective amount.
- the term "administration” means introducing the composition of the present invention to a patient by any suitable method, and the route of administration of the composition of the present invention is through various routes, either oral or parenteral, as long as it can reach the target tissue. Can be administered. Intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, oral administration, topical administration, intranasal administration, intrapulmonary administration, or rectal administration may be performed, but are not limited thereto.
- the method of the present invention may further include administering a protein (protein B) formed by self-assembly of a ferritin monomer to which a disease antigen epitope is fused to the individual.
- protein B protein formed by self-assembly of a ferritin monomer to which a disease antigen epitope is fused
- the disease antigen epitope may be a cancer antigen epitope, and may be a cancer antigen epitope specifically exemplified above.
- a protein (protein B) formed by self-assembly of a ferritin monomer to which a disease antigen epitope is fused may be administered simultaneously or sequentially with a ferritin protein to which a molecule capable of binding to an immune checkpoint molecule is fused.
- the order is not limited, and the protein made by self-assembly of ferritin monomers fused with disease antigen epitopes (Protein B) is the administration of ferritin protein (protein A) in which a molecule capable of binding to an immune checkpoint molecule is fused. It can be administered before or after administration.
- huHF is a globular protein (12 nm) composed of 24 monomers, and each monomer is composed of a total of 5 ⁇ -helix.
- the inventors of the present invention is the loop between each ⁇ -helix of the huHF monomer (AB loop among huHF 5T to 176G based on PDB 3AJO sequence; between 45D/46V, BC loop; 92D/93W, CD loop; 126D/127P, DE loop; 162E/163S )
- gp100 peptide which is one of the actual tumor antigens, was inserted at the N-terminus and C-terminus through gene cloning to obtain a delivery system in which the gp100 peptide was inserted at various positions of huHF (FIGS. 1 and 2 ).
- the present inventors have selected the surface composition of huHF nanoparticles with the best surveillance lymph node targeting efficiency as cancer-specific antigen delivery nanop
- the candidate proteins of Table 1 were subjected to PCR according to the vector schematic diagram of Table 2, and proteins huHF, huHF-gp100 (SEQ ID NO: 1; melanoma specific antigen), OVA (SEQ ID NO: 2), AH1 ( SEQ ID NO: 3) (AB; 45D/46V, BC; 92D/93W, CD; 126D/127P, DE; 162E/163S, N-terminal, C-terminal), huHF-PD1 (SEQ ID NO: 4; Active site of PD1 domain), huHF-TPP1 (SEQ ID NO: 5) (AB, CD loop), huHF- ⁇ PD-L1 HCDR3 (SEQ ID NO: 6) (CD loop, C-terminal) and huHF-smPD1 ( SEQ ID NO: 7) Particles were prepared.
- proteins huHF, huHF-gp100 SEQ ID NO: 1; melanoma specific antigen
- the OVA was used as an immunospecific antigen
- AH1 was used as a tumor specific antigen for colorectal cancer cells
- gp100 was used as a tumor specific antigen for melanoma cells. All the prepared plasmid expression vectors were purified on an agarose gel, and then the sequence was confirmed through complete DNA sequencing.
- PCR products required for preparation of each expression vector were sequentially inserted into the plasmid pT7-7 vector using the primer set in Table 3 to construct an expression vector capable of expressing each protein.
- it may further include a linker peptide of Table 4 below.
- E. coli strain BL21(DE3)[F-ompThsdSB(rB-mB-)] was transformed with the above-prepared expression vector, respectively, and ampicillin-resistant transformants were selected.
- the transformed E. coli was cultured in a flask (250 mL Erlenmeyer flasks, 37° C., 150 rpm) containing 50 mL of Luria-Bertani (LB) medium (containing 100 mg L-1 ampicillin).
- LB Luria-Bertani
- IPTG Isopropyl- ⁇ -Dthiogalactopyranosid
- the cultured E. coli was centrifuged at 4,500 rpm for 10 minutes to recover the cell precipitate and suspended in 5 ml of a disruption solution (10 mM Tris-HCl buffer, pH 7.5, 10 mM EDTA). Then, it was crushed using an ultrasonic crusher (Branson Ultrasonics Corp., Danbury, CT, USA). After crushing, centrifugation was performed at 13,000 rpm for 10 minutes, and the supernatant and insoluble aggregates were separated. The separated supernatant was used for later experiments.
- a disruption solution 10 mM Tris-HCl buffer, pH 7.5, 10 mM EDTA.
- the supernatant obtained in Example 2 was purified through a three-step process. First, 1) Ni2+-NTA affinity chromatography using the combination of histidine and nickel fused to the recombinant protein was performed, 2) the recombinant protein was concentrated and a fluorescent substance was attached through buffer exchange. Sucrose gradient ultracentrifugation was performed to separate only the attached self-assembled protein. Detailed description of each step is as follows.
- the cultured E. coli was recovered in the same manner as specified above, and the cell pellet was resuspended in 5 mL Lysis buffer (pH 8.0, 50 mM sodium phosphate, 300 mM NaCl, 20 mM imidazole), and an ultrasonic disruptor. Cells were disrupted. The crushed cell solution was centrifuged at 13,000 rpm for 10 minutes to separate only the supernatant, and then each recombinant protein was separated using a Ni2+-NTA column (Qiagen, Hilden, Germany) (washing buffer: pH 8.0, 50 mM sodium phosphate). , 300 mM NaCl, 80 mM imidazole / elution buffer: pH 8.0, 50 mM sodium phosphate, 300 mM NaCl, 200 mM imidazole).
- Lysis buffer pH 8.0, 50 mM sodium phosphate, 300 mM NaCl, 20 mM imidazole
- huHF-gp100 particles and huHF-PD1 particles were placed on a column with 5,000 g of 3 ml of recombinant protein eluted through Ni2 +- NTA affinity chromatography in an Ultracentrifugal filter (Amicon Ultra 100K, Millipore, Billerica, MA). Centrifugation was performed at 5,000 g until 1 ml of the solution remained. After that, to attach the NIR fluorescent substance cy5.5 and FITC (fluorescein isothiocyanate), the protein particles were buffered with sodium bicarbonate (0.1 M, pH 8.5) buffer, and the fluorescent substance at room temperature for 12 hours. Was attached.
- sucrose was added to PBS (2.7 mM KCl, 137 mM NaCl, 2 mM KH2PO4, 10 mM Na2HPO4, pH 7.4) buffer to contain 40%, 35%, 30%, 25%, 20% sucrose.
- PBS 2.7 mM KCl, 137 mM NaCl, 2 mM KH2PO4, 10 mM Na2HPO4, pH 7.4
- sucrose solution add 2 ml of sucrose solution at each concentration (45-20%) to the ultra-high-speed centrifugation tube (ultraclear 13.2 ml tube, Beckman), starting with the high-concentration solution, and then in the prepared buffer for self-assembly. After filling 1 ml of the present recombinant protein solution, ultra-high-speed centrifugation was performed at 4° C.
- TEM Transmission electron microscopy
- each of the particles formed spherical nanoparticles (FIGS. 3 and 5).
- each gp100-huHF-loops, huHF-PD1, huHF-TPP1 (AB, CD loops), huHF- ⁇ PD-L1 HCDR3 (CD loop, C-terminal), huHF-smPD1 particles through DLS (dynamic light scattering) measurement The diameter of the field was measured on the solution (Figs. 3 and 5).
- the present research team determined the binding ability of the purified recombinant protein of each protein (gp100-huHF-loops) produced in Example 3 to the transferrin receptor (TfR) in MST (Microscale Thermophoresis). ) Measured through a machine. As a result, it was confirmed that huHF nanoparticles containing no tumor antigen had the most excellent binding ability with TfR, and the binding ability of CD-loop-gp100 nanoparticles with tumor antigen inserted between CD helix was second. Through this, it was indirectly confirmed that the CD-loop-gp100 particles did not interfere with the binding to TfR most (FIG. 4).
- PD-1 Programmed cell death protein 1
- PD-L1 is a protein on the surface of T-cells. It binds to PD-L1, which is expressed on the surface of cancer cells, and induces decrease in T-cell activity. Therefore, when the binding site of PD-1 to bind to PD-L1 expressed on the surface of cancer cells is induced to inhibit the binding of PD-1 and PD-L1 in T cells by using the surface-expressed protein, T-cell activity is suppressed. It can be expected to increase the effectiveness of anticancer immunotherapy through the decrease.
- the PD-L1 binding site of PD-1 was synthesized in huHF (binding active site 22G-170V in the PD-1 sequence), PD-L1 targeting peptide TPP1, PD-L1 antibody HCDR3 sequence, the binding active site of PD-L1 (small PD1 domain)).
- the Langmuir equation was used to determine the binding capacity of PD-L1 antibody and huHF-PD1 protein and PD-L1, which are currently used immune antibody treatments. It was calculated using.
- the Kd value of huHF-PD1 and the recombinant protein PD-L1 was measured to be 327.59 nM, which is higher than 770 nM, which is a literature value of PD1-PDL1 binding affinity, which is similar to 255.10 nM, the Kd value of PD-L1 and PD-L1 antibodies. did. Through this, it was confirmed that the protein made by expressing the PD-1 binding domain on the huHF surface has the ability to bind to PD-L1 (FIG. 5).
- the binding capacity between the actually synthesized huHF- ⁇ PD-L1 HCDR3 (CD loop, C-terminal) protein and PD-L1 was also measured by ELISA, and the huHF- ⁇ PD-L1 HCDR3 (CD loop) particles were 71.24 nM, huHF- ⁇ PD-L1 HCDR3 (C-terminal) particles were measured to be 38.43 nM, respectively, confirming that these proteins also have a binding ability with PD-L1. (Fig. 5)
- the binding capacity of the huHF-TPP1 protein produced in Example 3 and PD-L1 was measured through a Microscale Thermophoresis (MST) machine.
- the Kd value of huHF-TPP1 (AB loop) with PD-L1 is 72.105 nM
- the Kd value of huHF-TPP1 (CD loop) with PD-L1 is 115.16 nM
- huHF- ⁇ PD-L1 HCDR3 (CD loop) ) was measured to be 71.24 nM
- huHF- ⁇ PD-L1 HCDR3 (C-terminal) was measured to be 38.43 nM (Fig. 5).
- the fluorescence signal was measured through a confocal (LSM 700) machine. It was confirmed that the binding ability of the CD-loop-gp100 protein with the tumor antigen inserted between the CD helix was superior to that of the huHF itself. Through this, it was also indirectly confirmed that the CD-loop-gp100 protein did not interfere with the binding of TfR most (FIG. 6).
- CT26 colorectal cancer cells and B16F10 melanoma cells were reacted with a protein at a concentration of 300 nM, and then the fluorescence signals were compared to confirm the cell uptake efficiency.
- huHF-PD1 ( Figure 7a), huHF- ⁇ PD-L1 HCDR3 (CD loops, C-terminal) ( Figure 7b), huHF-TPP1 (AB , CD loops) (Fig. 7c), huHF-smPD1 (Fig. 7c) protein was confirmed to exhibit a fluorescent signal by binding to cancer cells.
- PD-L1 antibody capable of masking PD-L1 expressed on the surface of cancer cells for 20 minutes
- huHF protein, huHF-PD1 protein, and huHF- ⁇ PD-L1 HCDR3 protein huHF-smPD1 protein were reacted respectively. When ordered, it was confirmed that neither was combined.
- the huHF protein and huHF-PD1 protein attached with a cy5.5 fluorescent substance were used in mice grown with CT-26 colorectal cancer cells.
- the PD-L1 antibody therapeutic agent that has been actually used in clinical practice was used as a control group.
- the particle targeting pattern in the body was observed with a Cy5.5 bandpass emission filater and a special Cmount lens or an IVIS spectrum imaging system (Caliper Life Sciences, Hopkinton, MA) (Fig. 9; right).
- the Y-axis represents the retention time in the body).
- the huHF-PD1 protein had better cancer cell targeting efficiency than the control huHF protein.
- the actual antibody treatment showed better cancer targeting efficiency and maintenance time in the body than the huHF-PD1 protein, but this is a result of the in vivo maintenance time of the antibody treatment being too long, which is directly related to the problem of immune side effects in the body. Therefore, it was confirmed that the protein according to the present invention has advantages in both side effects and side effects.
- PBS buffer
- huHF-gp100 loops protein By making PBS (buffer) and huHF-gp100 loops protein by the method of Examples 1 to 3, boosting the immune response of the immune cells in the lymph nodes by injecting the vaccine into C57BL/6 once a week for a total of 3 weeks, and then , The spleen where the immune cells gathered was excised and pulverized. After that, after extracting CD8+ T-cells in which an immune response was specifically induced by gp100 melanoma-specific antigen in the pulverized spleen, a specific partial antigen peptide of gp100 (KVPRNQDWL), which is known to induce an immune response in vitro.
- KVPRNQDWL a specific partial antigen peptide of gp100
- PBS buffer
- huHF-OVA loops protein By making PBS (buffer) and huHF-OVA loops protein by the method of Examples 1 to 3, boosting the immune response of the immune cells in the lymph nodes by injecting the vaccine into C57BL/6 once a week for a total of 3 weeks, and then , The spleen where the immune cells gathered was excised and pulverized. Then, in the pulverized spleen, the OVA immune peptide was used to identify a protein that best exposes the peptide to the dendritic cell surface by using an antibody that captures the surface-exposed dendritic cells (DC) through MHC-I.
- DC surface-exposed dendritic cells
- the nanoparticles containing the OVA peptide in the CD-loop induce the expression of the peptide surface on the MHC-I best, which disprove that it can most effectively activate the cytotoxic T cell activity in immunotherapy. to be.
- the B16F10 cell line was planted in each mouse and the growth rate of cancer was observed.
- the size of cancer cells was calculated by the following formula:
- the present inventors used a mouse Balb/c having a colon cancer tumor (CT26) of a certain size.
- CT26 colon cancer tumor
- PBS, PD-L1 antibody, and huHF-PD1 protein were injected intravenously at intervals of 3 days.
- CT26 colon cancer tumor
- the cancer-specific antigen epitopes (gp100 and AH1) were inserted into the CD-loop, which showed the best tumor growth inhibitory effect in mice with tumors of a certain size, and huHF-CD loop-gp100 and huHF-CD loop-AH1 ( 10 ⁇ M) protein was injected into mice at 3 days intervals by subcutaneous injection, and at the same time, huHF-PD1 (5 ⁇ M) and control PD-L1 antibody treatment samples were injected intravenously at 3 days intervals.
- the experiment using the huHF-CD loop-gp100 protein used C57BL/6 mice with B16F10 melanoma, and the experiment using the huHF-CD loop-AH1 protein used Balb/c mice with CT26 colon cancer. Each experiment used 5 mice per experimental group, and the size of cancer cells was calculated by the following formula:
- the experimental group was 1) no treatment group, 2) the first protein treatment group (AH1-huHF and gp100-huHF), 3) the antibody treatment group ( ⁇ -PD-L1), 4) the second protein Treatment group (huHF-PD1), 5) a group administered with a combination of a first protein and an antibody therapeutic agent (AH1-huHF+ ⁇ -PD-L1 and gp100-huHF+ ⁇ -PD-L1) and 6) a first protein and a second protein
- the combined administration groups (AH1-huHF+huHF-PD1 and gp100-huHF+huHF-PD1) were used.
- the present inventors In order to determine whether the huHF-PD1 protein is effective in cancer treatment through immune checkpoint suppression compared to the actual antibody treatment, PDL1 antibody, the present inventors have T-L1 antibody and huHF-PD1 protein react with cancer cells. The activity response of cells and the killing efficiency of cancer cells were compared.
- experimental group 1 No treat, 2) first protein treatment group (AH1-huHF and gp100-huHF), 3) antibody treatment group ( ⁇ -PD-L1), 4) second protein treatment group (huHF-PD1), 5) the combination administration group of the first protein and the antibody therapeutic agent (AH1-huHF+ ⁇ -PD-L1 and gp100-huHF+ ⁇ -PD-L1) and 6) the combination of the first protein and the second protein
- the active response of T-cells to the administration group (AH1-huHF+huHF-PD1 and gp100-huHF+huHF-PD1) was also observed, and as a result, experimental group 6 (AH1-huHF+huHF-PD1) with the best tumor growth inhibition result. And gp100-huHF+huHF-PD1), it was also confirmed that the T-cell activity is most excellent (FIG. 15C).
- huHF-PD1 protein has cancer treatment efficacy through immune checkpoint suppression compared to PDL1 antibody, which is an actual antibody treatment, and at the same time, the degree of induction of immune side effects when injected in vivo is also reduced.
- the biggest problem with the current antibody therapeutics is the problem of causing immune side effects due to long-term accumulation in the body when protein is injected, and the most representative cytokine that causes this immune side effect is known as IL-17. Accordingly, the present inventors performed an IL-17 detection test using the blood samples of Experimental Groups 1 to 6 described in Example 11.
- Example 11 As a result of the cancer growth inhibition test in Example 11, it was confirmed whether the first protein (CD loop-huHF) and the second protein (huHF-PD1) actually suppressed tumor growth in vivo and had a synergistic effect during combination treatment. Based on this, an experiment was conducted to see if the cancer recurs even after surgery.
- the experimental group was the same as in Example 11 1) no treatment group, 2) the first protein treatment group (AH1-huHF), 3) the antibody treatment group ( ⁇ -PD-L1), 4) agent 2 protein-treated group (huHF-PD1), 5) a group administered with a combination of the first protein and an antibody treatment (AH1-huHF+ ⁇ -PD-L1), and 6) a group administered with a combination of the first protein and a second protein (AH1-huHF+) huHF-PD1) was used.
- Example 11 An experiment was conducted to see whether cancer metastases even after surgery.
- the experimental group was the same as in Example 11 1) no treatment group, 2) the first protein treatment group (AH1-huHF), 3) the antibody treatment group ( ⁇ -PD-L1), 4) agent 2 protein-treated group (huHF-PD1), 5) a group administered with a combination of the first protein and an antibody treatment (AH1-huHF+ ⁇ -PD-L1), and 6) a group administered with a combination of the first protein and a second protein (AH1-huHF+) huHF-PD1) was used.
- mice were used.
- 5 mice were used per experimental group, and cancer metastasis was determined by extracting the lungs of mice used in all of the above experimental groups and counting cancer nodules (FIG. 17).
- the experimental group (1) no treatment group, 2) the first protein treatment group (AH1-huHF and gp100-huHF), 3) the antibody treatment group ( ⁇ -PD-L1) ), 4) the second protein treatment group (huHF-PD1), 5) the combination administration group of the first protein and antibody therapeutic agent (AH1-huHF+ ⁇ -PD-L1 and gp100-huHF+ ⁇ -PD-L1) and 6)
- the T-cell activity responses of the groups administered with the first protein and the second protein (AH1-huHF+huHF-PD1 and gp100-huHF+huHF-PD1) were observed.
- the protein was synthesized according to the method of Example 2, and the soluble and insoluble portions were confirmed according to the method of Example 18, which will be described later, and it was confirmed that the protein was self-assembled according to the method of Example 4.
- the binding force A of the prepared protein to transferrin was measured according to the following method.
- the reaction solution of each tube was put into the capillary of a Microscale thermophoresis device to obtain a homogeneous fluorescence intensity F cold without irradiation with a laser.
- the Microscale thermophoresis device (Monolith NT.115) was set to 40% MST power and the LED power so that the obtained fluorescence intensity value was within the range of 10,000 to 15,000, and irradiated with a laser for 30 seconds for each capillary in a heated state. The fluorescence intensity F hot was obtained.
- Various expression vectors based on pT7-7 were transformed into BL21 (DE3) competent cells.
- a single colony was inoculated into LB liquid medium (50 mL) to which 100 mg/L of ampicillin was added, and cultured at 37° C. and 130 rpm in a shaking incubator.
- the turbidity turbidity/optical density at 600 nm
- the expression of the target protein was induced through 1 mM IPTG administration.
- the cells in the culture medium were spun-down through centrifugation (13000 rpm, 10 minutes), and the cell pellet was collected and resuspended in 10 mM Tris-Hcl (pH 7.4) buffer. .
- Resuspended cells were ruptured using a Branson Sonifier (Branson Ultrasonics Corp., Danbury, CT). After sonication, the supernatant containing the soluble protein and the aggregates containing the insoluble protein were separated by centrifugation (13000 rpm, 10 minutes). The solubility was analyzed through SDS-PAGE analysis of the separated soluble and insoluble protein fractions. That is, the target protein bands stained with Coomassie were scanned with a densitometer (Duoscan T1200, Bio-Rad, Hercules, CA) and then the ratio of the water-soluble fraction was quantified. Specifically, using the scanned SDS-PAGE gel image, the ‘Quantity One’ program ‘Volume Rect. After setting the band thickness and background value with'Tool', the sum of the soluble and insoluble protein fractions was set to 100% using the'Volume Analysis Report' and the solubility was quantified.
- a Branson Sonifier Branson Ultrasonics Corp.,
- the protein was synthesized according to the method of Example 2, and the soluble and insoluble portions were confirmed according to the method of Example 19, and it was confirmed that the protein was self-assembled according to the method of Example 4.
- the binding force of the prepared protein to the transferrin receptor was measured according to the method of Example 17, and the concentration represented by Equation 1 was found to be 44.649 ⁇ 1.34 nM.
- the tumor inhibitory ability of the protein was evaluated according to the method of Example 11.
- the experimental group 1) PBS group, 2) antibody treatment group ( ⁇ -PD-L1), 3) first protein treatment group (huHF-PD1), 4) second protein treatment group (huHF-msmPD1) was used. I did.
- huHF is a substitution of some amino acids at the binding site with transferrin (exists in the BC loop), and a protein in which amino acids 81 and 83 in the sequence of SEQ ID NO: 1 are substituted with alanine was used.
- the binding force of the prepared protein to h-PD-L1 and m-PD-L1 was measured according to the method of Example 17, and the binding force to h-PD-L1 was 13.417 ⁇ 1.97 nM, to m-PD-L1. The binding force was found to be 177.14 ⁇ 3.32 nM.
- a protein was prepared in which an immune checkpoint molecule PD-L1 and a molecule binding to TIGIT were fused to ferritin, and its efficacy was confirmed.
- the HCDR3 sequence of the antibody was used, and the sequence used is shown in Table 10 below.
- the vector of Table 11 was prepared according to the method of Example 1, and at this time, the primer set of Table 12 was used.
- the protein was synthesized according to the method of Example 2.
- the tumor suppressing ability of the protein was determined by subcutaneous inoculation of a colon cancer cell line (CT26) into BALB/c mice and injecting the protein according to the schedule of FIG. 25, according to the method of Example 11. It was evaluated (FIG. 26).
- the experimental group 1) PBS group, 2) antibody treatment combined treatment group ( ⁇ -PD-L1, ⁇ -TIGIT), 3) protein treatment group (huHF-PD-L1-TIGIT dual blocker) was used.
- tumor tissues were removed for each treatment group and the weight was measured, and the results are shown in FIG. 27. From this, it is possible to confirm the excellent anticancer efficacy of the protein in which the molecule binding to PD-L1 and TIGIT is fused.
- a protein in which ⁇ -PD-L1 HCDR3 was fused to different positions of a ferritin monomer was prepared to confirm tumor suppression ability.
- a protein was prepared in which ⁇ -PD-L1 HCDR3 was fused to the AB loop, BC loop, CD loop, DE loop, and C-terminus (AB loop among huHF 5T to 176G based on PDB 3AJO sequence; between 45D/46V, BC loop; 92D/93W, CD loop; 126D/127P, DE loop; 162E/163S).
- This was prepared in the same manner as in Examples 1 and 2, except that the sequence of Table 10 was used.
- CT26 colorectal cancer cells at a concentration of 300 nM After reacting with the protein, the fluorescence signal was compared to confirm the cell uptake efficiency. It was confirmed that the huHF- ⁇ PD-L1 HCDR3 (AB, BC, CD, DE loops, C-terminal) protein bonded to the cancer cells and showed a fluorescent signal than the control huHF protein.
- Table 13 The sequence of Table 13 was used, and PCR was performed according to the vector schematic diagrams of FIGS. 29 to 36 and Table 14 below, and huHF- ⁇ PD1 HCDR3 (C-terminal), huHF- ⁇ CTLA4 HCDR3 (C-terminal), huHF ⁇ TIGIT HCDR3 (C-terminal), huHF- ⁇ LAG3 HCDR3 (C-terminal), huHF- ⁇ TIM3 HCDR3 (C-terminal), huHF- ⁇ PD-L1 HCDR3 (AB loop)- ⁇ TIGIT HCDR3 (C-terminal) (dual blocker) was prepared I did. All the prepared plasmid expression vectors were purified on an agarose gel, and then the sequence was confirmed through complete DNA sequencing.
- PCR products required for preparation of each expression vector were sequentially inserted into the plasmid pT7-7 vector using the primer set in Table 15 to construct an expression vector capable of expressing each protein nanoparticle.
- Insertion site 54 ⁇ _PD-L1_F C-terminal 55 ⁇ _PD-L1_R 56 ⁇ _PD1_F C-terminal 57 ⁇ _PD1_R 58 ⁇ _CTLA4_F C-terminal 59 ⁇ _CTLA4_R 60 ⁇ _LAG3_F C-terminal 61 ⁇ _LAG3-R 62 ⁇ _TIM3_F C-terminal 63 ⁇ _TIM3_R 64 ⁇ _TIGIT_F C-terminal 65 ⁇ _TIGIT_R 66 BC_ ⁇ _PD-L1_F BC loop 67 BC_ ⁇ _PD-L1_R
- Adhesion to the antigen was measured in the same manner as in Example 6, except that the antigen for each antibody was used.
- the binding power of the antibody is shown in Table 16, and the binding power of the protein of the example is shown in Tables 17 and 18. Referring to this, it can be seen that the proteins of the examples exhibit excellent binding power to human antigens.
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Abstract
Description
서열번호 | 후보 단백질 |
2 | Gp100 |
3 | Ovalbumin |
4 | AH1 |
5 | PD1 |
6 | TPP1 |
7 | αPD-L1 HCDR3 |
8 | smPD1 (small PD1 domain) |
9 | RFP |
단백질 | 발현 벡터 |
huHF | NH2-NdeI-(His)6-huHF-HindIII-COOH |
[gp100/AH1/OVA]-huHF loops | AB(45D/46V), BC(92D/93W), CD(126D/127P), DE(162E/163S):NH2-NdeI-(His)6-huHF-[gp100(KVPRNQDWL)/OVA(SIINFEKL)/AH1(SPSYVYHQF)]-huHF-HindIII-COOH
N-말단: NH2-NdeI-(His)6-[gp100(KVPRNQDWL)/OVA(SIINFEKL)/AH1(SPSYVYHQF)]-huHF-HindIII-COOH C-말단: NH2-NdeI-(His)6-huHF-[gp100(KVPRNQDWL)/OVA(SIINFEKL)/AH1(SPSYVYHQF)]-HindIII-COOH N-말단과 A helix 사이(6S/7Q), D helix 중간(156R/157K), E helix 중간(173H/174T), E helix와 C-말단 사이(178S/179D) NH2-NdeI-(His)6-[gp100(KVPRNQDWL)]-huHF-HindIII-COOH |
huHF-PD1 | NH2-NdeI-huHF-PD1(22-170)-HindIII-COOH |
huHF-TPP1 (AB, CD loop) | NH2-NdeI-huHF-linker-TPP1-linker-HindIII-COOH |
huHF-αPD-L1 HCDR3 (CD
loop, C-말단) |
NH2-NdeI-huHF-αPD-L1 HCDR3-HindIII-COOH |
huHF-smPD1 | NH2-NdeI-huHF-smPD1-HindIII-COOH |
huHF-RFP | NH2-NdeI-(His)6-huHF-Xho1-linker(G3SG3TG3SG3T)-RFP-HindIII-COOH |
서열번호 | 명칭 | 삽입부위 |
10 | N-gp100_F | N-말단 |
11 | N-gp100_R | |
12 | AB-gp100_F | AB loop |
13 | AB-gp100_R | |
14 | BC-gp100_F | BC loop |
15 | BC-gp100_R | |
16 | CD-gp100_F | CD loop |
17 | CD-gp100_R | |
18 | DE-gp100_F | DE loop |
19 | DE-gp100_R1 | |
20 | DE-gp100_R2 | |
21 | DE-gp100_R3 | |
22 | C-gp100_F | C-말단 |
23 | C-gp100_R | |
24 | NA-gp100_F_1 | N-말단과 A-helix 사이 |
25 | NA-gp100_F_2 | |
26 | NA-gp100_F_3 | |
27 | NA-gp100_R | |
28 | intD-gp100_F | D helix 중간 |
29 | intD-gp100_R_1 | |
30 | intD-gp100_R_2 | |
31 | intE-gp100_F | E helix 중간 |
32 | intE-gp100_R_1 | |
33 | intE-gp100_R_2 | |
34 | EC-gp100_F | E-helix와 C-말단 사이 |
35 | EC-gp100_R |
서열번호 | 명칭 |
36 | Linker1 |
37 | Linker2 |
38 | Linker3 |
TSA
삽입위치 |
TfR | gp100 |
N | Human | 18.484±1.13 |
Mouse | 48.323±2.86 | |
AB | Human | 29.663±0.71 |
Mouse | 40.787±2.85 | |
BC | Human | 14.043±3.27 |
Mouse | 48.021±3.37 | |
CD | Human | 4.8943±2.98 |
Mouse | 15.94±2.52 | |
DE | Human | 9.7809±1.97 |
Mouse | 30.485±1.11 | |
C | Human | 5.6533±1.33 |
Mouse | 34.795±0.98 | |
NA | Human | 152.29±2.68 |
EC | Human | 5.6768±1.29 |
D in | Human | 29.288±3.71 |
E in | Human | 6.276±1.76 |
TSA
삽입위치 |
TfR | AH1 |
N | Human | 84.648±0.83 |
AB | Human | 10.933±0.32 |
BC | Human | 106.64±0.74 |
CD | Human | 3.4503±3.49 |
DE | Human | 6.1146±4.11 |
C | Human | 5.3748±1.25 |
TSA
삽입위치 |
TfR | PD1 |
C | Human | 265.84±0.98 |
Mouse | 667.51±2.34 |
샘플 | TfR | 결합력 |
모델항원RFP-huHF
(TSA 삽입위치 C) |
Human | 127.23±0.71 |
WT huHF | Human | 2.498±1.77 |
Mouse | 7.59±2.72 |
단백질 | 수용성 분획 비율(%) | 단백질 | 수용성 분획 비율(%) |
N-OVA-huHF | 89.35 | N-gp100-huHF | 92.48 |
AB-OVA-huHF | 93.81 | AB-gp100-huHF | 93.86 |
BC-OVA-huHF | 95.74 | BC-gp100-huHF | 94.43 |
CD-OVA-huHF | 98.73 | CD-gp100-huHF | 95.57 |
DE-OVA-huHF | 96.82 | DE-gp100-huHF | 95.39 |
C-OVA-huHF | 96.62 | C-gp100-huHF | 96.40 |
N-AH1-huHF | 81.74 | NA-gp100-huHF | 67.22 |
AB-AH1-huHF | 94.63 | EC-gp100-huHF | 96.27 |
BC-AH1-huHF | 92.53 | D in-gp100-huHF | 48.21 |
CD-AH1-huHF | 98.47 | E in-gp100-huHF | 93.00 |
DE-AH1-huHF | 98.71 | PD1-huHF | 74.25 |
C-AH1-huHF | 87.90 | RFP-huHF | 98.31 |
서열번호 | 명칭 |
42 | huHF-αPD-L1* |
43 | huHF-αTIGIT* |
단백질 | 발현 벡터 |
huHF-PD-L1-TIGIT dual blocker | BC(92D/93W): NH2-NdeI-(His)6-huHF-[αTIGIT HCDR3]-huHF- αPD-L1 HCDR3-HindIII-COOH |
서열번호 | 명칭 | 삽입부위 |
44 | BC_α_PD-L1_F | BC loop |
45 | BC_α_PD-L1_R | |
46 | C_α_TIGIT_F | C-말단 |
47 | C_α_TIGIT_R |
서열번호 | 명칭 |
48 | huHF-αPD-L1 |
49 | huHF-αPD1 |
50 | huHF-αCTLA4 |
51 | huHF-αLAG3 |
52 | huHF-αTIM3 |
53 | huHF αTIGIT |
단백질 | 발현 벡터 |
huHF-αPD-L1 | NH2-NdeI-huHF-αPD-L1 HCDR3-HindIII-COOH |
huHF-αPD1 | NH2-NdeI-huHF-αPD1 HCDR3-HindIII-COOH |
huHF-αCTLA4 | NH2-NdeI-huHF-αCTLA4 HCDR3-HindIII-COOH |
huHF-αLAG3 | NH2-NdeI-huHF-αLAG3 HCDR3-HindIII-COOH |
huHF-αTIM3 | NH2-NdeI-huHF-αTIM3 HCDR3-HindIII-COOH |
huHF-αTIGIT | NH2-NdeI-huHF-αTIGIT HCDR3-HindIII-COOH |
huHF-PD-L1-TIGIT dual blocker | BC(92D/93W): NH2-NdeI-(His)6-huHF-[αTIGIT HCDR3]-huHF- αPD-L1 HCDR3-HindIII-COOH |
서열번호 | 명칭 | 삽입부위 |
54 | α_PD-L1_F | C-말단 |
55 | α_PD-L1_R | |
56 | α_PD1_F | C-말단 |
57 | α_PD1_R | |
58 | α_CTLA4_F | C-말단 |
59 | α_CTLA4_R | |
60 | α_LAG3_F | C-말단 |
61 | α_LAG3-R | |
62 | α_TIM3_F | C-말단 |
63 | α_TIM3_R | |
64 | α_TIGIT_F | C-말단 |
65 | α_TIGIT_R | |
66 | BC_α_PD-L1_F | BC loop |
67 | BC_α_PD-L1_R |
Human Ab | Kd (nM) | Catalog # |
αPD-L1 | 5.8227±0.52 | 10084-MM02
(Sino biological) |
αPD1 | 9.2096±1 | MBS154625(Mybiosource) |
αCTLA4 | 3.4228±1.81 | 11159-MM06(Sino biological) |
αTIM3 | 7.9993±1.01 | MBS4156568(Mybiosource) |
αTIGIT | 10.32±1.27 | MBS154627(Mybiosource) |
Sample | murine IC molecule
(standard) |
Kd
(nM) |
huHF-αPD-L1 | PD-L1 | 3.1692±2.56 |
huHF-αPD1 | PD1 | 87.889±2.47 |
huHF-αCTLA4 | CTLA4 | 17.513±0.462 |
huHF-αLAG3 | LAG3 | 37.817±1.82 |
huHF-αTIM3 | TIM3 | 7.0831±1.64 |
huHF-αTIGIT | TIGIT | 3.9997±2.29 |
huHF-α PD-L1-
αTIGIT |
PD-L1 | 4.9956±2.79 |
TIGIT | 4.7343±2.52 |
Sample | human IC molecule
(standard) |
Kd
(nM) |
huHF-αPD-L1 | PD-L1 | 10.708±4.52 |
huHF-αPD1 | PD1 | 17.776±4.38 |
huHF-αCTLA4 | CTLA4 | 10.167±3.11 |
huHF-αLAG3 | LAG3 | 18.498±3.17 |
huHF-αTIM3 | TIM3 | 13.03±4.21 |
huHF-αTIGIT | TIGIT | 7.1276±2.16 |
huHF-αPD-L1-αTIGIT | PD-L1 | 3.5605±1.07 |
TIGIT | 6.6423±3.46 |
Claims (20)
- 외부 표면에 면역 관문 분자와 결합 가능한 분자가 융합된 페리틴 단백질.
- 청구항 1에 있어서, 상기 면역 관문 분자와 결합 가능한 분자가 융합된 페리틴 단량체 24개가 자기 조립되어 이루어진 구형 단백질.
- 청구항 1에 있어서, 상기 면역 관문 분자와 결합 가능한 분자는 페리틴 단량체의 인접한 α-헬릭스들 사이 중 적어도 하나에 융합되는 단백질.
- 청구항 1에 있어서, 상기 면역 관문 분자와 결합 가능한 분자는 페리틴 단량체의 N-말단 또는 C-말단에 융합되는 단백질.
- 청구항 1에 있어서, 상기 면역 관문 분자와 결합 가능한 분자는 페리틴 단량체의 A-B루프, B-C루프, C-D루프 또는 D-E루프에 융합된 단백질.
- 청구항 1에 있어서, 상기 면역 관문 분자와 결합 가능한 분자는 페리틴 단량체의 N-말단과 A 헬릭스 사이 또는 E 헬릭스와 C-말단 사이에 융합된 단백질.
- 청구항 1에 있어서, 인간 트랜스페린 수용체에 대한 결합력이 떨어지도록 돌연 변이된 단백질.
- 청구항 1에 있어서, 상기 단백질을 구성하는 페리틴 단량체 중 적어도 하나가 서열번호 1의 서열에서 14번, 15번, 22번, 81번 또는 83번 아미노산이 알라닌, 글라이신, 발린 또는 류신으로 치환된 것인 단백질.
- 청구항 1에 있어서, 트랜스페린 수용체에 대한 결합력(K)이 다음 수학식 1을 만족하는 단백질:[수학식 1]K ≤ 700 nM(식 중, K = [P][T]/[PT]이고, 여기서 [P]는 상기 단백질과 상기 트랜스페린 수용체와의 결합 반응의 평형 상태에서의 상기 단백질의 농도를 나타내고, [T]는 상기 평형 상태에서의 상기 트랜스페린 수용체의 농도를 나타내며, [PT]는 상기 평형 상태에서의 상기 단백질과 상기 트랜스페린 수용체의 복합체의 농도를 나타냄).
- 청구항 9에 있어서, 상기 트랜스페린 수용체는 인간 트랜스페린 수용체인 단백질.
- 청구항 1에 있어서, 상기 면역 관문 분자는 Her-2/neu, VISTA, 4-1BBL, Galectin-9, Adenosine A2a receptor, CD80, CD86, ICOS, ICOSL, BTLA, OX-40L, CD155, BCL2, MYC, PP2A, BRD1, BRD2, BRD3, BRD4, BRDT, CBP, E2F1, MDM2, MDMX, PPP2CA, PPM1D, STAT3, IDH1, PD1, CTLA4, PD-L1, PD-L2, LAG3, TIM3, TIGIT, BTLA, SLAMF7, 4-1BB, OX-40, ICOS, GITR, ICAM-1, BAFFR, HVEM, LFA-1, LIGHT, NKG2C, SLAMF7, NKp80, LAIR1, 2B4, CD2, CD3, CD16, CD20, CD27, CD28, CD40L, CD48, CD52, EGFR family, AXL, CSF1R, DDR1, DDR2, EPH receptor family, FGFR family, VEGFR family, IGF1R, LTK, PDGFR family, RET, KIT, KRAS, NTRK1 및 NTRK2으로 이루어진 군에서 선택되는 어느 하나인 단백질.
- 청구항 1에 있어서, 상기 면역 관문 분자와 결합 가능한 분자는 상기 면역 관문 분자에 대한 리간드, 항체 또는 이의 단편인 단백질.
- 청구항 1에 있어서, 상기 페리틴은 인간 페리틴 중쇄인 단백질.
- 청구항 1에 있어서, 상기 단백질은 대장균 생산 시스템에서 수용성 분획 비율이 40% 이상으로 존재하는 단백질.
- 청구항 1 내지 14 중 어느 한 항의 단백질을 포함하는 암의 치료 또는 예방용 약학 조성물.
- 청구항 15에 있어서, 상기 암은 뇌암, 두경부암, 방광암, 유방암, 자궁경부암, 결장암, 결장직장암, 자궁내막암, 식도암, 백혈병, 폐암, 간암, 난소암, 췌장암, 전립선암, 직장암, 신장암, 위암, 고환암, 자궁암, 혈관 종양, 편평세포암종, 선암종, 소세포 암종, 흑색종, 신경교종, 신경아세포종, 육종, 후두암, 이하선암, 담도암, 갑상선암, 광선각화증, 급성 림프구성 백혈병, 급성 골수 백혈병, 샘낭암종, 선종, 선 평편상피암종, 항문관암, 항문암, 항문직장암, 성상세포종, 큰질어귀샘암, 기저세포 암종, 담즙암, 골암, 골수암, 기관지암, 기관지샘 암종, 카시노이드, 담관암종, 만성 림프구성 백혈병, 만성 골수성 백혈병, 투명세포 암종, 결합조직암, 낭선종, 소화계통암, 십이지장암, 내분비계암, 내배엽동종양, 자궁내막증식증, 자궁내막모양 선암종, 내피세포암, 뇌실막세포, 상피세포암, 안와암, 국소결절성 과증식, 담낭암, 날문방암, 위 기저부 암, 가스트린종, 교모세포종, 글루카곤종, 심장암, 혈관아세포종, 혈관내피종, 혈관종, 간샘종, 간 선종증, 간담도암, 간세포 암종, 호지킨병, 회장암, 인슐린종, 상피내 신생물, 상피내 편평세포 신생물, 간내 담도암, 침윤성 편평세포암종, 공장암, 관절암, 골반암, 거대 세포 암종, 대장암, 림프종, 악성 중피세포 종양, 수아세포종, 수질상피종, 뇌막암, 중피암, 전이성 암종, 구강암, 점막표피모양 암종, 다발성 골수종, 근육암, 비강관암, 신경계암, 비-상피 피부암, 비-호지킨 림프종, 연맥 세포 암종, 핍지교종암, 구강암, 골육종, 유두상 장액성 선암종, 음경암, 인두암, 뇌하수체 종양, 형질세포종, 가육종, 폐 아세포종, 직장암, 신세포 암종, 호흡계 암, 망막아세포종, 장액성 암종, 부비강암, 피부암, 소세포 암종, 소장암, 평활근육암, 연조직암, 소마토스타틴-분비 종양, 척추암, 편평세포암종, 선조 근육암, 중피세포하층암, T 세포 백혈병, 설암, 요관암, 요도암, 자궁경부암, 자궁몸통암, 질암, VIPoma, 외음부암, 고분화 암종 및 윌름 종양으로 이루진 군에서 선택된 것인 암의 치료 또는 예방용 약학 조성물.
- 청구항 15에 있어서, 질환 항원 에피토프가 융합된 페리틴 단량체가 자기 조립되어 이루어지고, 트랜스페린 수용체에 대한 결합력(K)이 다음 수학식 4를 만족하는 단백질을 더 포함하는 암의 치료 또는 예방용 약학 조성물:[수학식 4]K ≤ 700 nM(식 중, K = [P][T]/[PT]이고, 여기서 [P]는 상기 단백질과 상기 트랜스페린 수용체와의 결합 반응의 평형 상태에서의 상기 단백질의 농도를 나타내고, [T]는 상기 평형 상태에서의 상기 트랜스페린 수용체의 농도를 나타내며, [PT]는 상기 평형 상태에서의 상기 단백질과 상기 트랜스페린 수용체의 복합체의 농도를 나타냄).
- 청구항 15에 있어서, 상기 질환 항원 에피토프는 gp100, MART-1, Melna-A, MAGE-A3, MAGE-C2, Mammaglobin-A, proteinsase-3, mucin-1, HPV E6, LMP2, PSMA, GD2, hTERT, PAP, ERG, NA17, ALK, GM3, EPhA2, NA17-A, TRP-1, TRP-2, NY-ESO-1, CEA, CA 125, AFP, Survivin, AH1, ras, G17DT, MUC1, Her-2/neu, E75, p53, PSA, HCG, PRAME, WT1, URLC10, VEGFR1, VEGFR2, E7, Tyrosinase 펩타이드, B16F10, EL4 또는 신생항원(neoantigen)인 암의 치료 또는 예방용 약학 조성물.
- 청구항 17에 있어서, 2종의 단백질이 병용 투여되기 위한 암의 치료 또는 예방용 약학 조성물.
- 청구항 17에 있어서, 2종의 단백질이 동시, 개별 또는 순차적으로 투여되는 암의 치료 또는 예방용 약학 조성물.
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Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR20150088597A (ko) * | 2014-01-24 | 2015-08-03 | 국립대학법인 울산과학기술대학교 산학협력단 | 항원 펩타이드-페리틴 융합단백질 및 이를 포함하는 백신 조성물 |
KR20170047120A (ko) * | 2015-10-22 | 2017-05-04 | 고려대학교 산학협력단 | 암 특이적 에피토프와 연결된 단백질 나노입자 및 이를 포함하는 암 면역치료용 조성물 |
KR20180008353A (ko) * | 2016-07-15 | 2018-01-24 | 한국과학기술연구원 | 신규 나노케이지 및 그의 용도 |
KR20190115262A (ko) * | 2018-04-02 | 2019-10-11 | 충남대학교산학협력단 | 페리틴 자가조립체에 항원 펩타이드 및 면역 증강제가 결합된 나노 입자 및 이의 용도 |
Family Cites Families (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2014123399A1 (ko) * | 2013-02-08 | 2014-08-14 | 경북대학교 산학협력단 | 인간 페리틴 유래 융합폴리펩티드 |
EP3512563A1 (en) * | 2016-09-16 | 2019-07-24 | The Johns Hopkins University | Protein nanocages with enhanced mucus penetration for targeted tissue and intracellular delivery |
KR20200123132A (ko) * | 2018-02-21 | 2020-10-28 | 아지노모토 가부시키가이샤 | 융합 단백질 |
-
2020
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- 2020-11-02 JP JP2022525447A patent/JP2023500293A/ja active Pending
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Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR20150088597A (ko) * | 2014-01-24 | 2015-08-03 | 국립대학법인 울산과학기술대학교 산학협력단 | 항원 펩타이드-페리틴 융합단백질 및 이를 포함하는 백신 조성물 |
KR20170047120A (ko) * | 2015-10-22 | 2017-05-04 | 고려대학교 산학협력단 | 암 특이적 에피토프와 연결된 단백질 나노입자 및 이를 포함하는 암 면역치료용 조성물 |
US10206987B2 (en) | 2015-10-22 | 2019-02-19 | Korea University Research And Business Foundation | Protein nanoparticle linked with cancer specific epitope and composition for cancer immunotherapy comprising the same |
KR20180008353A (ko) * | 2016-07-15 | 2018-01-24 | 한국과학기술연구원 | 신규 나노케이지 및 그의 용도 |
KR20190115262A (ko) * | 2018-04-02 | 2019-10-11 | 충남대학교산학협력단 | 페리틴 자가조립체에 항원 펩타이드 및 면역 증강제가 결합된 나노 입자 및 이의 용도 |
Non-Patent Citations (1)
Title |
---|
LEE B.-R. ET AL: "Engineered Human Ferritin Nanoparticles for Direct Delivery of Tumor Antigens to Lymph Node and Cancer Immunotherapy", SCIENTIFIC REPORTS, vol. 6, 11 October 2016 (2016-10-11), pages 1 - 12, XP002792566, DOI: 10.1038/srep35182 * |
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