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KR-102962848-B1 - Two-component toxin-loaded antibody-drug conjugate and its uses

KR102962848B1KR 102962848 B1KR102962848 B1KR 102962848B1KR-102962848-B1

Abstract

The present disclosure provides an antibody-drug conjugate loaded with a two-component toxin. By linking MMAF in series with another drug unit at the cysteine binding site on the antibody, the two exhibit a significant synergistic effect, which can effectively improve the effect of killing tumor cells. This provides a novel solution for the development of high-efficiency and low-toxicity ADCs.

Inventors

  • 후앙 창지앙
  • 순 유시앙
  • 시옹 지우카이
  • 콩 나나
  • 얀 신신

Assignees

  • 맙플렉스 인터내셔널 컴퍼니 리미티드

Dates

Publication Date
20260511
Application Date
20210719

Claims (20)

  1. Antibody-drug conjugate having the following chemical formula I or a pharmaceutically acceptable salt thereof: In the above formula, Ab is an antibody or an antigen-binding fragment, wherein the antigen-binding fragment is one or more antibody fragments that retain the ability to specifically bind to an antigen; S is a sulfur atom in the sulfhydryl residue formed after the opening of the interchain disulfide bond on Ab; Aa is an amino acid unit comprising one or more amino acids, wherein the amino acid unit Aa is -glycine-, -alanine-, -valine-, -leucine-, -isoleucine-, -proline-, -phenylalanine-, -tryptophan-, -methionine-, -tyrosine-, -serine-, -threonine-, -cysteine-, -asparagine-, -glutamine-, -aspartic acid-, -glutamic acid-, -lysine-, -arginine-, -histidine-, -citrulline-, -lysine(trityl)-, -lysine(monomethoxycitrate)-, -lysine(fluorenylmethoxycarbonyl)-, -valine-citrulline-, -valine-alanine-, -valine-lysine-, -valine-lysine(trityl)-, -valine-lysine(monomethoxycitrate)-, Selected from the group consisting of -valine-lysine (fluorenyl methoxycarbonyl)-, -valine-arginine-, -phenylalanine-citrulline-, -phenylalanine-lysine-, -phenylalanine-lysine (trityl)-, -phenylalanine-lysine (monomethoxycitrateyl)-, -phenylalanine-lysine (fluorenyl methoxycarbonyl)-, -leucine-citrulline-, -isoleucine-citrulline-, -phenylalanine-arginine-, -phenylalanine-arginine-arginine-, -glycine-glycine-phenylalanine-glycine-, -glycine-phenylalanine-leucine-glycine-, and -alanine-leucine-alanine-leucine-; G is a cracking unit, selected from the following structural formulas: and (In the above formula, R1 and R2 are H, C1 - C10 alkyl and It is independently selected from a group consisting of D 1 is MMAF and; D2 is MK4827, ceritinib, chemical formula [ A second active drug unit selected from the group consisting of Dxd, SN38, and exatecan having ]; p is an integer selected from 1, 2, 3, 4, 5, 6, 7, and 8.
  2. In paragraph 1, Ab Intact antibodies of murine antibodies, chimeric antibodies, humanized antibodies, or fully human antibodies; or The antigen-binding fragment of the above intact antibody Phosphorus, antibody-drug conjugate, or pharmaceutically acceptable salt thereof.
  3. In paragraph 2, The antigen-binding fragment is selected from the group consisting of Fab, Fab', F(ab) 2 and F(ab') 2 ; The antibody is selected from the group consisting of bispecific antibodies and multispecific antibodies, Antibody-drug conjugate or pharmaceutically acceptable salt thereof.
  4. In paragraph 3, An antibody-drug conjugate or a pharmaceutically acceptable salt thereof, wherein Ab is a human IgG1 or human IgG4 antibody, or a form having an Fc domain or a hinge domain having partial amino acid mutations, substitutions, and deletions thereof.
  5. In paragraph 1, An antibody-drug conjugate or a pharmaceutically acceptable salt thereof, wherein the first active drug unit D1 is selected from the group consisting of the structure of the following chemical formula, its tautomers, mesomes, racemic mixtures, enantiomers and diastereomers, mixtures thereof, and pharmaceutically acceptable salts or solvates thereof: .
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  8. In paragraph 1, Antibody-drug conjugates or pharmaceutically acceptable salts thereof selected from the structures of the following chemical formulas: In the above formula, p is an integer selected from 1, 2, 3, 4, 5, 6, 7, and 8.
  9. In any one of paragraphs 1 through 5 and paragraph 8, Antibody-drug conjugates or pharmaceutically acceptable salts thereof for use in the treatment or prevention of cancer.
  10. Intermediate compound having the structure of the following chemical formula II: In the above formula, Aa is an amino acid unit comprising one or more amino acids, wherein the amino acid unit Aa is -glycine-, -alanine-, -valine-, -leucine-, -isoleucine-, -proline-, -phenylalanine-, -tryptophan-, -methionine-, -tyrosine-, -serine-, -threonine-, -cysteine-, -asparagine-, -glutamine-, -aspartic acid-, -glutamic acid-, -lysine-, -arginine-, -histidine-, -citrulline-, -lysine(trityl)-, -lysine(monomethoxycitrate)-, -lysine(fluorenylmethoxycarbonyl)-, -valine-citrulline-, -valine-alanine-, -valine-lysine-, -valine-lysine(trityl)-, -valine-lysine(monomethoxycitrate)-, Selected from the group consisting of -valine-lysine (fluorenyl methoxycarbonyl)-, -valine-arginine-, -phenylalanine-citrulline-, -phenylalanine-lysine-, -phenylalanine-lysine (trityl)-, -phenylalanine-lysine (monomethoxycitrateyl)-, -phenylalanine-lysine (fluorenyl methoxycarbonyl)-, -leucine-citrulline-, -isoleucine-citrulline-, -phenylalanine-arginine-, -phenylalanine-arginine-arginine-, -glycine-glycine-phenylalanine-glycine-, -glycine-phenylalanine-leucine-glycine-, and -alanine-leucine-alanine-leucine-; G is a cracking unit, selected from the following structural formulas: and (In the above formula, R1 and R2 are H, C1 - C10 alkyl and It is independently selected from a group consisting of D 1 is MMAF and; D2 is MK4827, ceritinib, chemical formula [ It is a second active drug unit selected from the group consisting of Dxd, SN38, and exatecan having ].
  11. In Paragraph 10, A compound comprising a first active drug unit D1 selected from the group consisting of the structure of the following chemical formula, its tautomers, mesomes, racemic mixtures, enantiomers and diastereomers, mixtures thereof, and pharmaceutically acceptable salts or solvates thereof: .
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  13. In Paragraph 10, Compounds selected from the structures of the following chemical formulas: .
  14. In any one of paragraphs 10, 11 and 13, A compound for use in the treatment or prevention of cancer.
  15. A method for synthesizing a compound according to paragraph 10, wherein the synthesis route of the said synthesis method And; The above synthesis method Step 1: Compound Fmoc-Aa-G, the second active drug unit D2 , and organic base 1 are dissolved in an appropriate amount of organic solvent 1, and the same is brought into contact and reacted to form compound Fmoc-Aa- GD2. The obtaining stage, Step 2: A step of obtaining compound Aa- GD2 by removing the protecting group Fmoc from the compound Fmoc-Aa- GD2 , and Step 3: Dissolving the above compound Aa- GD2 , compound Mc- D1 (wherein Mc is maleimidocaproyl), and condensation agent 1 in an appropriate amount of organic solvent 2, and contacting and reacting them to obtain compound Mc- D1 -Aa- GD2. A synthesis method comprising, wherein the organic base 1 is selected from one or more of N,N-diisopropylethylamine, triethylamine, and pyridine; the organic solvent 1 and the organic solvent 2 are each independently selected from one or both of DMF and DMA; and the condensation agent 1 is selected from one or more of TSTU, HATU, HBTU, HCTU, PyBop, CDMT, and T3P.
  16. In paragraph 15, The compound is selected from the structures set forth in paragraph 13; synthetic path Selected from, Synthesis method.
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Description

Two-component toxin-loaded antibody-drug conjugate and its uses The present disclosure relates to the field of antibody-drug conjugates, and in particular to antibody-drug conjugates loaded with a two-component toxin via an antibody. An antibody-drug conjugate (ADC) refers to a biological drug in which a biologically active drug is linked to an antibody via a chemical linker. An ADC is akin to a precision-guided weapon system where the biologically active drug acts as a killing weapon that precisely attacks diseased cells under the guidance of the antibody. Therefore, ADCs combine the dual advantages of the high efficiency of cytotoxic drug molecules and the high targeting of antibodies. As of March 2021, only 11 antibody-conjugated drugs have been approved worldwide (as shown in Table 1), all of which are ADC molecules in which a single target antibody is bound to a single toxin molecule. Due to the limited number of antigens on the surface of tumor cells, the level of antigen expression required for effective ADC activity varies depending on the characteristics of the different antigens. ADCs require at least 10⁴ antigens/cell to ensure the delivery of a lethal dose of cytotoxic drug. Ideally, the antigen targeted by the antibody portion of the ADC should be uniformly expressed on the surface of tumor cells and have a high copy number (greater than 10⁵ /cell). However, the reality is that typically only a limited number of antigens (about 5,000 to 10⁶ antigens/cell) exist on the surface of tumor cells, and the ideal average DAR value of an ADC is 3.5 to 4 (for example, the average DAR value of Brentuximab vedotin is 4, and the average DAR value of Trastuzumab emtansine is 3.5), and the amount of drug delivered to tumor cells is very small, so the drug effect on tumor cells is poor, and this is also considered one of the main causes of clinical failure of ADCs. Antibody-drug conjugates currently on the marketDrug generic namecompanyTargetSales periodBrentuximab Vedotin (Brentuximab vedotin)Seattle, Takeda (Seattle, Takeda)CD30Release/2011Ado-trastuzumab emtansine (Ado-trastuzumab emtansine)Genentech (Genentech)Her2Release/2013Inotuzumab ozogamicin (Inotuzumab ozogamicin)Pfizer (Pfizer)CD22Release/2017Gemtuzoumab ozogamicin (Gemtuzumab ozogamicin)PfizerCD33Release/2017Moxetumomab Pseudotox-tdfk (Moxetumomab pasudotox-tdfk)AstraZeneca (AstraZeneca)CD22Release/2018Enfortumab Vedotin-ejfv (Enfortumab vedotin-ejfv)Seattle/Astellas (Seattle/Astellas)Nectin-4Release/2019Folatuzumab Vedotin-piiq (Polatuzumab vedotin-piiq)GenentechCD79bRelease/2019Pharm-Trastuzumab Deluxecan-nxki (Fam-trastuzumab deruxtecan-nxki)AstraZeneca/Daiichi SankyoHer2Release/2019Sacituzumab govitecan-hziy (Sacituzumab govitecan-hziy)Immunomedics (Immunomedics)Trop2Release/2020Belantamab Mapodotin (Belantamab mafodotin)GlaxoSmithKline (Ireland) Limited (Glaxosmithkline (Ireland) Ltd)BCMARelease/2020Cetuximab Sarotalocan) (Cetuximab sarotalocanRakuten Medical (Rakuten Medical)EGFRRelease/2020Note: Mylotarg was approved on the list in 2000, withdrawn from the market in 2010, and re-approved on the list in 2017. To solve the above problems, the present disclosure provides an antibody-drug conjugate capable of loading a two-component toxin. In particular, the present disclosure provides an antibody-drug conjugate comprising the structure of the following formula I: [Chemical Formula I] In the above formula, Ab is an antibody or antigen-binding fragment; S is a sulfur atom in the sulfhydryl residue formed after the opening of the interchain disulfide bond on Ab; Aa is an amino acid unit containing one or more amino acids; G is an arbitrary crack unit; D 1 is MMAF and; D2 is a second active drug unit that is not MMAF; p is an integer selected from 1, 2, 3, 4, 5, 6, 7, and 8. The antibody or antigen-binding fragment Ab is reactive with an antigen or its epitope associated with cancer, an infectious disease, or an autoimmune disease. Optionally, Ab is an intact antibody, fragment (e.g., Fab, Fab', F(ab) 2 , and F(ab') 2 ), or subfragment (e.g., a single-strand structure) derived from a murine, chimeric, primate, humanized, or human monoclonal antibody, and said antibody includes bispecific and multispecific antibodies. Optionally, Ab is an intact form of a murine, chimeric, primate, humanized, or human monoclonal antibody, and said Ab has an Fc domain and a hinge domain of a form of a human IgG1 or human IgG4 antibody, or a form having partial amino acid mutations, substitutions, or deletions thereof. The first active drug unit D1 is selected from auristatin cytotoxic agents; preferably, the first active drug unit D1 contains a free carboxyl group. In some preferred embodiments, the first drug unit D1 is selected from the structure of the following formula, and its tautomers, mesomes, racemics, enantiomers, diastereomers, or mixtures thereof, and pharmaceutically acceptable salts or solvates thereof: . The second active drug unit D2 is selected from