Composite or salt thereof, and method for producing the same
Patent Information
- Authority / Receiving Office
- JP · JP
- Patent Type
- Patents
- Current Assignee / Owner
- AJINOMOTO CO INC
- Filing Date
- 2022-03-15
- Publication Date
- 2026-06-23
AI Technical Summary
【0019】 本発明によれば、抗体およびフェリチン粒子の比率を高度に制御することができる。また、抗体に対するフェリチン粒子中の連結部位、およびフェリチン粒子に対する抗体中の連結部位の双方において位置選択性を実現することができる。したがって、本発明は、極めて高度な品質管理を実現することができる。 また、天然型ヒトフェリチン単量体を含むヒトフェリチン粒子、および/またはペプチドを含まないリンカーを使用することで、ヒトへの臨床応用に際して、免疫原性を回避することができる。 さらに、本発明の複合体またはその塩において、抗体およびフェリチン粒子は、それぞれ、薬物および/または標的化剤としての機能を担うことができる。これは、抗体自体が薬物および/または標的化剤として使用でき、また、フェリチン粒子についても、薬物送達手段および/または標的化剤として使用できるためである。例えば、フェリチン粒子は、内腔中に多量の物質を封入可能であり、また、表面上のフェリチン単量体に薬物を結合可能であるため、薬物送達手段として使用できる。特に、薬物が生体中で分解し易い性質を有する場合(例、siRNA等の核酸分子)、フェリチンは、内腔中に多量の物質を封入して、生体から物質を隔離できるので、薬物送達手段として有用である。また、フェリチン粒子は、特定臓器または組織(例、リンパ節、肝臓、脾臓、癌組織)に集積し易く、また、血液脳関門(BBB)を透過し易いため、標的化剤として使用できる。したがって、本発明の複合体またはその塩は、薬物、薬物送達手段、および/または標的化剤としての機能を、抗体およびヒトフェリチン粒子の一方または双方に振り分けることができるので、医薬または試薬等の製品として有用である。
Smart Images
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Abstract
Claims
1. A complex or salt thereof comprising (A) one IgG antibody, and (B) two human ferritin particles each containing one or more human ferritin H chains linked thereto.
2. The complex or a salt thereof according to claim 1, wherein the human ferritin particles are 24-mers of human ferritin H chains.
3. The complex or salt thereof according to claim 1 or 2, wherein the human ferritin H chain is a natural human ferritin H chain that may have a methionine residue at position 1 deleted.
4. The aforementioned composite is given by the following formula (1): X1-L1-Y-L2-X2 (1) [During the ceremony, X1 and X2 each represent human ferritin particles containing one or more human ferritin H chains. Y is an IgG antibody, L1 and L2 represent linkers, respectively. Two human ferritin particles, represented by X1 and X2, are linked to two linkers, represented by L1 and L2, via atoms in the side chains of amino acid residues located at the same positions in the human ferritin H chains contained therein. The complex or a salt thereof according to any one of claims 1 to 3.
5. The complex or a salt thereof according to claim 4, wherein the linker does not contain a peptide portion.
6. The complex or salt thereof according to claim 4 or 5, wherein the atom in the side chain of the amino acid residue located at the same position in the human ferritin H chain is a sulfur atom in the side chain of the cysteine residue.
7. The complex or salt thereof according to claim 6, wherein the cysteine residue is located at position 91 and / or 103, corresponding to the reference position of the natural human ferritin H chain.
8. The complex or a salt thereof according to any one of claims 1 to 7, wherein the antibody is a chimeric monoclonal antibody, a humanized monoclonal antibody, or a human monoclonal antibody.
9. The complex or salt thereof according to any one of claims 4 to 7, wherein the IgG antibody represented by Y is linked to two linkers represented by L1 and L2 via atoms in the side chains of amino acid residues located at the same positions in the two heavy chain constant regions.
10. The complex or salt thereof according to claim 9, wherein the atoms in the side chains of amino acid residues located at the same position in two heavy chain constant regions are nitrogen atoms in the side chains of lysine residues.
11. The complex or salt thereof according to claim 10, wherein the lysine residue is located at position 246 / 248, 288 / 290, or 317 of the IgG antibody heavy chain according to EU numbering.
12. The complex or a salt thereof according to any one of claims 1 to 11, wherein human ferritin particles containing one or more human ferritin H chains contain one or more modified human ferritin H chains and one or more unmodified human ferritin H chains.
13. The complex or salt thereof according to any one of claims 1 to 12, wherein the human ferritin particles are particles containing each of the following human ferritin H chains (a) to (c) in the following average number per particle: (a) Number of modified human ferritin H chains linked to the linker: 1; (b) The number of modified human ferritin H chains that are not linked to a linker and have modified groups introduced: n; and (c) Number of unmodified human ferritin H chains: m; Here, n is a number between 0 and 23, m is a number between 0 and 23, and the sum of n and m is 23.
14. The complex or salt thereof according to any one of claims 1 to 13, wherein the human ferritin particles are particles containing an average of one or more unmodified human ferritin H chains.
15. The complex or salt thereof according to any one of claims 1 to 14, wherein the human ferritin particles are particles containing an average of 12 or more unmodified human ferritin H chains.
16. The complex or a salt thereof according to claim 13, wherein the modifying group includes a reactive group.
17. The complex or salt thereof according to claim 16, wherein the reactive group is a bioorthogonal functional group to a protein.
18. The complex or salt thereof according to claim 17, wherein the bioorthogonal functional group to the protein comprises one or more substructures selected from the group consisting of maleimide moiety, azide moiety, ketone moiety, aldehyde moiety, thiol moiety, alkene moiety, alkyne moiety, halogen moiety, tetrazine moiety, nitrone moiety, hydroxylamine moiety, nitrile moiety, hydrazine moiety, boronic acid moiety, cyanobenzothiazole moiety, allyl moiety, phosphine moiety, disulfide moiety, thioester moiety, α-halocarbonyl moiety, isonitrile moiety, cydonone moiety, and selenium moiety.
19. The composite or salt thereof according to claim 13, wherein the modifying group contains a functional substance.
20. The complex or salt thereof according to claim 19, wherein the functional substance comprises one or more parts selected from the group consisting of peptides, proteins, nucleic acids, low molecular weight organic compounds, chelators, glycans, lipids, high molecular weight polymers, and metals.
21. A composite or salt thereof according to any one of claims 1 to 20, wherein human ferritin particles have a substance in their lumen.
22. A method for producing a composite or a salt thereof, The complex comprises (A) one IgG antibody and (B) two human ferritin particles each containing one or more human ferritin H chains linked thereto. Method including (1) to (3) below: (1) To produce a modified IgG antibody containing two primary reactive groups by introducing a primary reactive group into an IgG antibody; (2) To produce modified particles containing one or more modified human ferritin H chains containing the second reactive group by introducing a second reactive group that can react with a first reactive group into human ferritin particles containing one or more human ferritin H chains containing the second reactive group; and (3) Reacting the modified IgG antibody with the modified particles to obtain the complex.
23. The aforementioned composite is given by the following formula (1): X1-L1-Y-L2-X2 (1) [During the ceremony, X1 and X2 each represent human ferritin particles containing one or more human ferritin H chains. Y is an IgG antibody, L1 and L2 represent linkers, respectively. The method according to claim 22, wherein two human ferritin particles, represented by X1 and X2, are linked to two linkers, represented by L1 and L2, via atoms in the side chains of amino acid residues located at the same positions in the human ferritin H chain contained therein.
24. The method according to claim 23, wherein the linker does not contain a peptide portion.
25. The method according to claim 23 or 24, wherein the atom in the side chain of the amino acid residue located at the same position in the human ferritin H chain is a sulfur atom in the side chain of the cysteine residue.
26. The method according to claim 25, wherein the cysteine residue is located at position 91 and / or 103, corresponding to the reference position of the natural human ferritin H chain.
27. A method for producing a composite or a salt thereof, The complex comprises (A) one IgG antibody, and (B) two human ferritin particles each containing one or more modified human ferritin H chains and one or more unmodified human ferritin H chains linked thereto. Method including (1) to (4) below: (1) To produce a modified IgG antibody containing two primary reactive groups by introducing a primary reactive group into an IgG antibody; (2) To produce first modified particles containing two or more modified human ferritin H chains containing the second reactive group by introducing a second reactive group that can react with a first reactive group into human ferritin particles containing two or more human ferritin H chains containing the second reactive group; (3) In the first modified particle, a portion of the two or more modified human ferritin H chains is replaced with an unmodified human ferritin H chain to produce a second modified particle containing one or more modified human ferritin H chains and one or more unmodified human ferritin H chains; and (4) Reacting the modified IgG antibody with the second modified particle to obtain the complex.
28. A method for producing a composite or a salt thereof, The complex comprises (A) one IgG antibody, and (B) two human ferritin particles each containing one or more modified human ferritin H chains and one or more unmodified human ferritin H chains linked thereto. Method including (1) to (4) below: (1) To produce an IgG antibody containing two primary reactive groups by introducing a primary reactive group into an IgG antibody; (2) To produce first modified particles containing two or more modified human ferritin H chains containing the second reactive group by introducing a second reactive group that can react with a first reactive group into human ferritin particles containing two or more human ferritin H chains containing the second reactive group; (3) Reacting the modified IgG antibody with the first modified particle to produce a complex or a salt thereof comprising (A') one IgG antibody and (B') two human ferritin particles each containing two or more modified human ferritin H chains linked thereto; and (4) A complex or salt thereof comprising (A') one IgG antibody and (B') two human ferritin particles each containing two or more modified human ferritin H chains linked thereto, wherein a portion of the two or more modified human ferritin H chains is replaced with an unmodified human ferritin H chain to obtain a complex or salt thereof comprising (A) one IgG antibody and (B) two human ferritin particles each containing one or more modified human ferritin H chains and one or more unmodified human ferritin H chains linked thereto.
29. The aforementioned composite is given by the following formula (1): X1-L1-Y-L2-X2 (1) [During the ceremony, X1 and X2 each represent human ferritin particles containing one or more modified human ferritin H chains and one or more unmodified human ferritin H chains, respectively. Y is an IgG antibody, L1 and L2 represent linkers, respectively. The method according to claim 27 or 28, represented by the following: Two human ferritin particles, represented by X1 and X2, are linked to two linkers, represented by L1 and L2, via atoms in the side chains of amino acid residues located at the same positions in the human ferritin H chains contained therein.
30. The method according to claim 29, wherein the linker does not contain a peptide portion.
31. The method according to claim 30, wherein the atom in the side chain of the amino acid residue located at the same position in the human ferritin H chain is a sulfur atom in the side chain of the cysteine residue.
32. The method according to claim 31, wherein the cysteine residue is located at position 91 and / or 103, corresponding to the reference position of the natural human ferritin H chain.
33. The method according to any one of claims 22 to 32, wherein the first reactive group and the second reactive group are bioorthothic functional groups for a protein.
34. The bioorthogonal functional group to the protein comprises one or more substructures selected from the group consisting of maleimide moiety, azide moiety, ketone moiety, aldehyde moiety, thiol moiety, alkene moiety, alkyne moiety, halogen moiety, tetrazine moiety, nitrone moiety, hydroxylamine moiety, nitrile moiety, hydrazine moiety, boronic acid moiety, cyanobenzothiazole moiety, allyl moiety, phosphine moiety, disulfide moiety, thioester moiety, α-halocarbonyl moiety, isonitrile moiety, cydonone moiety, and selenium moiety. The method according to claim 33, wherein the first reactive group and the second reactive group are selected in a combination that can react with each other.