WO2022078524A2 - Specific conjugation of an antibody - Google Patents

Specific conjugation of an antibody Download PDF

Info

Publication number
WO2022078524A2
WO2022078524A2 PCT/CN2021/128453 CN2021128453W WO2022078524A2 WO 2022078524 A2 WO2022078524 A2 WO 2022078524A2 CN 2021128453 W CN2021128453 W CN 2021128453W WO 2022078524 A2 WO2022078524 A2 WO 2022078524A2
Authority
WO
WIPO (PCT)
Prior art keywords
antibody
independently
alkyl
och
protein
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/CN2021/128453
Other languages
English (en)
French (fr)
Other versions
WO2022078524A3 (en
WO2022078524A4 (en
Inventor
Robert Zhao
Qingliang YANG
Xiaolei Liu
Lingli Zhang
Yuanyuan Huang
Wenjun Li
Hangbo YE
Juan Wang
Huihui GUO
You Zhou
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Hangzhou Dac Biotech Co Ltd
Original Assignee
Hangzhou Dac Biotech Co Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Hangzhou Dac Biotech Co Ltd filed Critical Hangzhou Dac Biotech Co Ltd
Priority to JP2024526535A priority Critical patent/JP2024541058A/ja
Priority to CA3236930A priority patent/CA3236930A1/en
Priority to KR1020247017868A priority patent/KR20240095442A/ko
Priority to EP21879565.6A priority patent/EP4426727A2/en
Priority to CN202180099689.6A priority patent/CN117980327A/zh
Priority to PCT/CN2021/128453 priority patent/WO2022078524A2/en
Priority to AU2021362997A priority patent/AU2021362997A1/en
Publication of WO2022078524A2 publication Critical patent/WO2022078524A2/en
Publication of WO2022078524A3 publication Critical patent/WO2022078524A3/en
Priority to KR1020247017883A priority patent/KR20240095316A/ko
Priority to JP2024526537A priority patent/JP2024541059A/ja
Priority to AU2022383265A priority patent/AU2022383265A1/en
Priority to PCT/CN2022/123901 priority patent/WO2023078021A1/en
Priority to CA3236852A priority patent/CA3236852A1/en
Priority to CN202280066606.8A priority patent/CN118434765A/zh
Priority to EP22889047.1A priority patent/EP4426741A4/en
Priority to TW113142378A priority patent/TW202506738A/zh
Priority to TW111141166A priority patent/TWI864472B/zh
Priority to EP22889295.6A priority patent/EP4426729A1/en
Priority to AU2022381163A priority patent/AU2022381163A1/en
Priority to CN202280069631.1A priority patent/CN118215676A/zh
Priority to JP2024526538A priority patent/JP2024542073A/ja
Priority to CA3236754A priority patent/CA3236754A1/en
Priority to PCT/CN2022/129122 priority patent/WO2023078273A1/en
Publication of WO2022078524A4 publication Critical patent/WO2022078524A4/en
Priority to TW112138435A priority patent/TWI923934B/zh
Priority to ZA2024/03371A priority patent/ZA202403371B/en
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6835—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site
    • A61K47/6851—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site the antibody targeting a determinant of a tumour cell
    • A61K47/6867—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site the antibody targeting a determinant of a tumour cell the tumour determinant being from a cell of a blood cancer
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6801—Drug-antibody or immunoglobulin conjugates defined by the pharmacologically or therapeutically active agent
    • A61K47/6803—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates
    • A61K47/68037—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates the drug being a camptothecin [CPT] or derivatives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6801—Drug-antibody or immunoglobulin conjugates defined by the pharmacologically or therapeutically active agent
    • A61K47/6803—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6801—Drug-antibody or immunoglobulin conjugates defined by the pharmacologically or therapeutically active agent
    • A61K47/6803—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates
    • A61K47/68031—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates the drug being an auristatin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6801—Drug-antibody or immunoglobulin conjugates defined by the pharmacologically or therapeutically active agent
    • A61K47/6803—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates
    • A61K47/68033—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates the drug being a maytansine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6801—Drug-antibody or immunoglobulin conjugates defined by the pharmacologically or therapeutically active agent
    • A61K47/6803—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates
    • A61K47/68035—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates the drug being a pyrrolobenzodiazepine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6801—Drug-antibody or immunoglobulin conjugates defined by the pharmacologically or therapeutically active agent
    • A61K47/6803—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates
    • A61K47/6811—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates the drug being a protein or peptide, e.g. transferrin or bleomycin
    • A61K47/6817—Toxins
    • A61K47/6831—Fungal toxins, e.g. alpha sarcine, mitogillin, zinniol or restrictocin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6835—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site
    • A61K47/6851—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site the antibody targeting a determinant of a tumour cell
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6889—Conjugates wherein the antibody being the modifying agent and wherein the linker, binder or spacer confers particular properties to the conjugates, e.g. peptidic enzyme-labile linkers or acid-labile linkers, providing for an acid-labile immuno conjugate wherein the drug may be released from its antibody conjugated part in an acidic, e.g. tumoural or environment
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2878—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95

Definitions

  • the present invention relates to a process for preparing a homogeneous conjugate of an antibody or antibody-like protein molecule/agent via linkage of certain sulphurs of cysteine sites in the antibody.
  • the present invention also relates to methods of making the conjugates in a specific manner comprising either generation of specific thiols of an antibody or antibody-like protein agent, followed by reaction with drug/linker complexes, or generation of specific thiols of an antibody or antibody-like protein agent and conjugation of a synthetic linker-drug assembly to the protein molecule simultaneously in one pot reaction. It also relates to methods of using the homogeneous conjugate in targeted prophylaxis or treatment of cancer, infection and immunological disorders.
  • NMMs next generation maleimides
  • dibromopyri-dazinediones A. Maruani, et al, Nat. Commun., 2015, 6, 6645; M.T. Lee, et al, Chem.
  • Zinc amino complexes have more advantages over ZnCl 2 in coordination of reduction of disulfide bond in an antibody.
  • zinc amino complexes are much bulkier than ZnCl 2 and can be more 3-D space selectivly to be inserted in certain positions (e.g.
  • zinc amino complexes are more stable in a water based solution, for instance, the stability constant of zinc ammonia complex ion is 2.9 x 10 9 (https: //chempedia. info/info/stability_constants/) , which in turn, slow the precipitation in a neutral pH phosphate buffer.
  • the conjugation strategy of this invention has robust manufacturability to yield highly homogeneous ADCs without antibody engineering and can successfully tackle an important shortcoming in current ADC preparation methods.
  • This conjugation strategy can be applied directly to other antibody likes of proteins.
  • the resulting homogeneous ADCs demonstrate improved pharmacokinetics, superior efficacy, and reduced toxicity in vivo compared to analogous conventional heterogeneous ADCs.
  • the present invention provides conjugation process with improved homogeneity of an antibody conjugate, or antibody-like protein conjugate, in particular, an antibody –drug conjugate (ADC) , wherein over 75%of payloads (drugs) are specifically conjugated to the disulfide bond sites between heavy-light chains of an antibody.
  • ADC antibody –drug conjugate
  • the homogenous conjugation process comprises the following three key steps:
  • PBS Mes, Bis-Tris, Bis-Tris Propane, Pipes, Aces, Mopso, Bes, Mops, Hepes, Tes, Pipps, Dipso, Tapso, Heppso, Tris-up, Tris-HCl, Tricine, Hepps, Gly-Gly, Bicine, Taps, Hepee, Acetates, Histidine, Citrates, MES, or Borates, etc. ) to selectively reduce interchain disulfide bonds within the antibody, or antibody-like protein to generate thiols;
  • step (b) introducing an effective amount of linker or payload/linker complex/assembly bearing thiol reactive groups (e.g., a drug containing maleimide terminal) to react with the thiol groups resulted from step (a) ; and
  • oxidant e.g. dehydroascorbic acid (DHAA)
  • DHAA dehydroascorbic acid
  • step (d) can be replaced by: adding an effective amount of cystine or relative disulfide compound to quench the unreacted reductant, while generating cysteine from the reduction of the cystine to quench the excessive conjugation linker or linker/payload complex containing thiol reactive groups (e.g. maleimide) .
  • (NR 1 R 2 R 3 ) m1 can be form a dimer, trimer, tetramer, pentamer, or hexamer wherein these polymers are covalently linked among N, R 1 , R 2 and R 3 ; and N, R 1 , R 2 or R 3 themselve can form heterocyclic, carbocyclic, diheterocyclic, or dicarbocyclic rings.
  • the transition metal cation-amino chelate/complex, M (NR 1 R 2 R 3 ) m1 m2+ , used in step (a) is 0.01 mM –1.0 mM in concentration, or 0.5 ⁇ 20 equivalents in moles of the protein, and it can be added to the reaction solution with a water-soluble organic solvent, selected from, ethanol, methanol, propanol, propandiol, DMA, DMF, DMSO, THF, CH 3 CN.
  • a water-soluble organic solvent selected from, ethanol, methanol, propanol, propandiol, DMA, DMF, DMSO, THF, CH 3 CN.
  • the reductant is an organic phosphine, preferably selected from Tris (2-carboxyethyl) phosphine (TECP) or Tris (hydroxypropyl) phosphine and its use in the reaction solution is 0.02 mM –1.0 mM in concentration, or 1.0 –20 equivalents in moles of the protein.
  • the oxidant to be added in step (c) may be DHAA, Fe 3+ , I 2 , Cu 2+ , Mn 3+ , MnO 2 , or mixture of Fe 3+ /I - .
  • the oxidant used in the reaction solution is 0.02 mM -1.0 mM in concentration, or 1 -100 equivalents in moles of the protein.
  • the optimum pH in the conjugation reaction is typically between about 5.0 to 8.0, and preferably, about 5.5 to 7.5.
  • the optimum temperature in the conjugation reaction is typically between about -5 to about 40 °C, and preferably, about 0 to 37 °C; more preferably about 2 to 8 °C.
  • the optimum time of the conjugation reaction is typically between about 15 min to about 48 preferably, about 30 min to overnight (10 ⁇ 16 h) .
  • the optimal reaction conditions e.g. pH, temeperature, buffer, concentrations of the reactants
  • the optimal reaction conditions e.g. pH, warmth, buffer, concentrations of the reactants
  • the optimal reaction conditions e.g. pH, warmth, buffer, concentrations of the reactants
  • the optimal reaction conditions e.g. pH, warmth, buffer, concentrations of the reactants of course are depended upon specifically an antibody-like protein, a payload/linker complex, a reductant and/or M (NR 1 R 2 R 3 ) m1
  • the antibody or antibody-like protein in the conjugation process can be any types of antibodies or proteins as long as they have two or more disulfide bonds in the protein for differentiation of reduction.
  • the payload/linker complex may be any types or formats as long as it has a thiol reactive group.
  • the ADCs prepared by the process of the present application have more than 80%of payloads conjugated in the Fab region of an antibody, in contrast to the conventional process wherein around 40%of the payloads are in the Fab region of an antibody and about 70%of the payloads are in the Fab region of an antibody using the process of WO2020164561.
  • Figure 1 The proposed mechanism that zinc amino complexes coordinate the reduction of the disulfide bonds in an antibody.
  • FIG. 1 Middle-level characterization of ADC after N-deglycosylation and reduction.
  • FIG. 4 The Percentage of Drug Loaded Peptides which were generated with hydrolases from the BCMA conjugate C-408b and analysized with UPLC-MS.
  • (a) Light chain (LC) Peptide [GEC] with zero or one drug molecule attached (D0 and D1)
  • Heavy chain (HC) Peptide [SCDK] at the arm with zero or one drug molecule attached (D0 and D1)
  • (X here is an amino acid that will be disclosed in a separated patent application) .
  • the results demonstrated the payloads were conjugated mainly (over 85%) at the cysteine sites between the light-heavy chains of the antibody.
  • the figure indicates that all the 9 conjugates had antitumor activity, and the conjugate C-408b prepared with the method of this invention had better in vivo activity than that prepared by the regular method.
  • the figure indicates that all the 9 conjugates had antitumor activity, and the conjugate C-408b prepared with the method of this invention had better in vivo activity than that prepared by the regular method.
  • Alkyl refers to an aliphatic hydrocarbon group or univalent groups derived from alkane by removal of one or two hydrogen atoms from carbon atoms. It may be straight or branched having C 1 -C 8 (1 to 8 carbon atoms) in the chain. “Branched” means that one or more lower C numbers of alkyl groups such as methyl, ethyl or propyl are attached to a linear alkyl chain.
  • Exemplary alkyl groups include methyl, ethyl, n-propyl, i-propyl, n-butyl, t-butyl, n-pentyl, 3-pentyl, octyl, nonyl, decyl, cyclopentyl, cyclohexyl, 2, 2-dimethylbutyl, 2, 3-dimethylbutyl, 2, 2-dimethylpentyl, 2, 3-dimethylpentyl, 3, 3-dimethylpentyl, 2, 3, 4-trimethylpentyl, 3-methyl-hexyl, 2, 2-dimethylhexyl, 2, 4-dimethylhexyl, 2, 5-dimethylhexyl, 3, 5-dimethylhexyl, 2, 4-dimethylpentyl, 2-methylheptyl, 3-methylheptyl, n-heptyl, isoheptyl, n-octyl, and isooctyl.
  • a C 1 -C 8 alkyl group can be unsubstituted or substituted with one or more groups including, but not limited to, -C 1 -C 8 alkyl, -O- (C 1 -C 8 alkyl) , -aryl, -C (O) R', -OC (O) R', -C (O) OR', -C (O) NH 2 , -C (O) NHR', -C (O) N (R') 2 , -NHC (O) R', -SR', -S (O) 2 R', -S (O) R', -OH, -halogen, -N 3 , -NH 2 , -NH (R') , -N (R') 2 and -CN; where each R' is independently selected from -C 1 -C 8 alkyl and aryl.
  • Halogen refers to fluorine, chlorine, bromine or iodine atom; preferably fluorine and chlorine atom.
  • Heteroalkyl refers to C 2 -C 8 alkyl in which one to four carbon atoms are independently replaced with a heteroatom from the group consisting of O, S and N.
  • Carbocycle refers to a saturated or unsaturated ring having 3 to 8 carbon atoms as a monocycle or 7 to 13 carbon atoms as a bicycle.
  • Monocyclic carbocycles have 3 to 6 ring atoms, more typically 5 or 6 ring atoms.
  • Bicyclic carbocycles have 7 to 12 ring atoms, arranged as a bicycle [4, 5] , [5, 5] , [5, 6] or [6, 6] system, or 9 or 10 ring atoms arranged as a bicycle [5, 6] or [6, 6] system.
  • Representative C 3 -C 8 carbocycles include, but are not limited to, -cyclopropyl, -cyclobutyl, -cyclopentyl, -cyclopentadienyl, -cyclohexyl, -cyclohexenyl, -1, 3-cyclohexadienyl, -1, 4-cyclohexadienyl, -cycloheptyl, -1, 3-cycloheptadienyl, -1, 3, 5-cycloheptatrienyl, -cyclooctyl, and -cyclooctadienyl.
  • a “C 3 -C 8 carbocycle” refers to a 3-, 4-, 5-, 6-, 7-or 8-membered saturated or unsaturated nonaromatic carbocyclic ring.
  • a C 3 -C 8 carbocycle group can be unsubstituted or substituted with one or more groups including, but not limited to, -C 1 -C 8 alkyl, -O- (C 1 -C 8 alkyl) , -aryl, -C (O) R', -OC (O) R', -C (O) OR', -C (O) NH 2 , -C (O) NHR', -C (O) N (R') 2 , -NHC (O) R', -SR', -S (O) R', -S (O) 2 R', -OH, -halogen, -N 3 , -NH 2 , -NH (R') , -N (R') 2 and
  • Alkenyl refers to an aliphatic hydrocarbon group containing a carbon-carbon double bond which may be straight or branched having 2 to 8 carbon atoms in the chain.
  • alkenyl groups include ethenyl, propenyl, n-butenyl, i-butenyl, 3-methylbut-2-enyl, n-pentenyl, hexylenyl, heptenyl, octenyl.
  • Alkynyl refers to an aliphatic hydrocarbon group containing a carbon-carbon triple bond which may be straight or branched having 2 to 8 carbon atoms in the chain.
  • exemplary alkynyl groups include ethynyl, propynyl, n-butynyl, 2-butynyl, 3-methylbutynyl, 5-pentynyl, n-pentynyl, hexylynyl, heptynyl, and octynyl.
  • Alkylene refers to a saturated, branched or straight chain or cyclic hydrocarbon radical of 1-18 carbon atoms, and having two monovalent radical centers derived by the removal of two hydrogen atoms from the same or two different carbon atoms of a parent alkane.
  • Typical alkylene radicals include, but are not limited to: methylene (-CH 2 -) , 1, 2-ethyl (-CH 2 CH 2 -) , 1, 3-propyl (-CH 2 CH 2 CH 2 -) , 1, 4-butyl (-CH 2 CH 2 CH 2 CH 2 -) , and the like.
  • Alkenylene refers to an unsaturated, branched or straight chain or cyclic hydrocarbon radical of 2-18 carbon atoms, and having two monovalent radical centers derived by the removal of two hydrogen atoms from the same or two different carbon atoms of a parent alkene.
  • Alkynylene refers to an unsaturated, branched or straight chain or cyclic hydrocarbon radical of 2-18 carbon atoms, and having two monovalent radical centers derived by the removal of two hydrogen atoms from the same or two different carbon atoms of a parent alkyne.
  • Typical alkynylene radicals include, but are not limited to: acetylene, propargyl and 4-pentynyl.
  • Aryl or Ar refers to an aromatic or hetero aromatic group, composed of one or several rings, comprising three to fourteen carbon atoms, preferentially six to ten carbon atoms.
  • hetero aromatic group refers one or several carbon on aromatic group, preferentially one, two, three or four carbon atoms are replaced by O, N, Si, Se, P or S, preferentially by O, S, and N.
  • Heterocycle refers to a ring system in which one to four of the ring carbon atoms are independently replaced with a heteroatom from the group of O, N, S, Se, B, Si and P. Preferable heteroatoms are O, N and S. Heterocycles are also described in The Handbook of Chemistry and Physics, 78th Edition, CRC Press, Inc., 1997-1998, p. 225 to 226, the disclosure of which is hereby incorporated by reference.
  • Preferred nonaromatic heterocyclic include epoxy, aziridinyl, thiiranyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, oxiranyl, tetrahydrofuranyl, dioxolanyl, tetrahydropyranyl, dioxanyl, dioxolanyl, piperidyl, piperazinyl, morpholinyl, pyranyl, imidazolinyl, pyrrolinyl, pyrazolinyl, thiazolidinyl, tetrahydrothiopyranyl, dithianyl, thiomorpholinyl, dihydropyranyl, tetrahydropyranyl, dihydropyranyl, tetrahydropyridyl, dihydropyridyl, tetrahydropyrimidinyl, dihydrothiopyranyl, azepanyl, as well as the fused
  • heteroaryl refers to a 3 to 14, preferably 5 to 10 membered aromatic hetero, mono-, bi-, or multi-cyclic ring.
  • examples include pyrrolyl, pyridyl, pyrazolyl, thienyl, pyrimidinyl, pyrazinyl, tetrazolyl, indolyl, quinolinyl, purinyl, imidazolyl, thienyl, thiazolyl, benzothiazolyl, furanyl, benzofuranyl, 1, 2, 4-thiadiazolyl, isothiazolyl, triazolyl, tetrazolyl, isoquinolyl, benzothienyl, isobenzofuryl, pyrazolyl, carbazolyl, benzimidazolyl, isoxazolyl, pyridyl-N-oxide, as well as the fused systems resulting from the condensation with a phenyl group
  • Alkyl “, “cycloalkyl “, “alkenyl “, “alkynyl “, “aryl “, “heteroaryl “, “heterocyclic” and the like refer also to the corresponding “alkylene “, “cycloalkylene “, “alkenylene “, “alkynylene “, “arylene “, “heteroarylene “, “heterocyclene” and the likes which are formed by the removal of two hydrogen atoms.
  • Arylalkyl refers to an acyclic alkyl radical in which one of the hydrogen atoms bonded to a carbon atom, typically a terminal or sp 3 carbon atom, is replaced with an aryl radical.
  • Typical arylalkyl groups include, benzyl, 2-phenylethan-1-yl, 2-phenylethen-1-yl, naphthylmethyl, 2-naphthylethan-1-yl, 2-naphthylethen-1-yl, naphthobenzyl, 2-naphthophenylethan-1-yl and the like.
  • Heteroarylalkyl refers to an acyclic alkyl radical in which one of the hydrogen atoms bonded to a carbon atom, typically a terminal or sp 3 carbon atom, is replaced with a heteroaryl radical.
  • heteroarylalkyl groups are 2-benzimidazolylmethyl, 2-furylethyl.
  • Examples of a “hydroxyl protecting group” include, methoxymethyl ether, 2-methoxyethoxymethyl ether, tetrahydropyranyl ether, benzyl ether, p-methoxybenzyl ether, trimethylsilyl ether, triethylsilyl ether, triisopropylsilyl ether, t-butyldimethylsilyl ether, triphenylmethylsilyl ether, acetate ester, substituted acetate esters, pivaloate, benzoate, methanesulfonate and p-toluenesulfonate.
  • leaving group refers to a functional group that can be substituted by another functional group.
  • Such leaving groups are well known in the art, and examples include, a halide (e.g., chloride, bromide, and iodide) , methanesulfonyl (mesyl) , p-toluenesulfonyl (tosyl) , trifluoro-methylsulfonyl (triflate) , and trifluoromethylsulfonate.
  • a preferred leaving group is selected from nitrophenol; N-hydroxysuccinimide (NHS) ; phenol; dinitrophenol; pentafluorophenol; tetrafluorophenol; difluorophenol; monofluorophenol; pentachlorophenol; triflate; imidazole; dichlorophenol; tetrachlorophenol; 1-hydroxybenzotriazole; tosylate; mesylate; 2-ethyl-5-phenylisoxazolium-3′-sulfonate, anhydrides formed its self, or formed with the other anhydride, e.g. acetyl anhydride, formyl anhydride; or an intermediate molecule generated with a condensation reagent for peptide coupling reactions or for Mitsunobu reactions.
  • NHS N-hydroxysuccinimide
  • Boc tert-butoxy carbonyl
  • BroP bromotrispyrrolidinophosphonium hexafluorophosphate
  • CDI 1, 1'-carbonyldiimidazole
  • DCC dicyclohexylcarbodiimide
  • DCE dichloroethane
  • DCM dichloromethane
  • DIAD diisopropylazodicarboxylate
  • DIBAL-H diisobutyl-aluminium hydride
  • DIPEA diisopropylethylamine
  • DEPC diethyl phosphorocyanidate
  • DMA N, N-dimethyl acetamide
  • DMAP 4- (N, N-dimethylamino) pyridine
  • DMF N, N-dimethylformamide
  • DMSO dimethylsulfoxide
  • DTT dithiothreitol
  • EDC 1- (3-dimethylamino)
  • amino acid (s) can be natural and/or unnatural amino acids, preferably alpha-amino acids.
  • Natural amino acids are those encoded by the genetic code, which are alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tyrosine. tryptophan and valine.
  • the unnatural amino acids are derived forms of proteinogenic amino acids.
  • Examples include hydroxyproline, lanthionine, 2-aminoisobutyric acid, dehydroalanine, gamma-aminobutyric acid (the neurotransmitter) , ornithine, citrulline, beta alanine (3-aminopropanoic acid) , gamma-carboxyglutamate, selenocysteine (present in many noneukaryotes as well as most eukaryotes, but not coded directly by DNA) , pyrrolysine (found only in some archaea and one bacterium) , N-formylmethionine (which is often the initial amino acid of proteins in bacteria, mitochondria, and chloroplasts) , 5-hydroxytryptophan, L-dihydroxyphenylalanine, triiodothyronine, L-3, 4-dihydroxyphenylalanine (DOPA) , and O-phosphoserine.
  • DOPA 4-dihydroxyphenylalanine
  • amino acid also includes amino acid analogs and mimetics.
  • Analogs are compounds having the same general H 2 N (R) CHCO 2 H structure of a natural amino acid, except that the R group is not one found among the natural amino acids. Examples of analogs include homoserine, norleucine, methionine-sulfoxide, and methionine methyl sulfonium.
  • an amino acid mimetic is a compound that has a structure different from the general chemical structure of an alpha-amino acid but functions in a manner similar to one.
  • the term “unnatural amino acid” is intended to represent the “D” stereochemical form, the natural amino acids being of the “L” form.
  • amino acid sequence is then preferably a cleavage recognition sequence for a protease.
  • Many cleavage recognition sequences are known in the art. See, e.g., Matayoshi et al. Science 247: 954 (1990) ; Dunn et al. Meth. Enzymol. 241: 254 (1994) ; Seidah et al. Meth. Enzymol. 244: 175 (1994) ; Thornberry, Meth. Enzymol. 244: 615 (1994) ; Weber et al. Meth. Enzymol. 244: 595 (1994) ; Smith et al. Meth. Enzymol.
  • sequence is selected from the group consisting of Val-Cit, Ala-Val, Val-Ala-Val, Lys-Lys, Ala-Asn-Val, Val-Leu-Lys, Cit-Cit, Val-Lys, Ala-Ala-Asn, Lys, Cit, Ser, and Glu.
  • “Pharmaceutically” or “pharmaceutically acceptable” refer to molecular entities and compositions that do not produce an adverse, allergic or other untoward reaction when administered to an animal, or a human, as appropriate.
  • “Pharmaceutically acceptable solvate” or “solvate” refer to an association of one or more solvent molecules and a disclosed compound.
  • solvents that form pharmaceutically acceptable solvates include, but are not limited to, water, isopropanol, ethanol, methanol, DMSO, ethyl acetate, acetic acid and ethanolamine.
  • “Pharmaceutically acceptable excipient” includes any carriers, diluents, adjuvants, or vehicles, such as preserving or antioxidant agents, fillers, disintegrating agents, wetting agents, emulsifying agents, suspending agents, solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents and the like.
  • preserving or antioxidant agents such as preserving or antioxidant agents, fillers, disintegrating agents, wetting agents, emulsifying agents, suspending agents, solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents and the like.
  • the use of such media and agents for pharmaceutical active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active ingredient, its use in the therapeutic compositions is contemplated. Supplementary active ingredients can also be incorporated into the compositions as suitable therapeutic combinations.
  • pharmaceutically acceptable salts refer to derivatives of the disclosed compounds wherein the parent compound is modified by making acid or base salts thereof.
  • the pharmaceutically acceptable salts include the conventional non-toxic salts or the quaternary ammonium salts of the parent compound formed, for example, from non-toxic inorganic or organic acids.
  • such conventional non-toxic salts include those derived from inorganic acids such as hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, nitric and the like; and the salts prepared from organic acids such as acetic, propionic, succinic, tartaric, citric, methanesulfonic, benzenesulfonic, glucuronic, glutamic, benzoic, salicylic, toluenesulfonic, oxalic, fumaric, maleic, lactic and the like.
  • Further addition salts include ammonium salts such as tromethamine, meglumine, epolamine, etc., metal salts such as sodium, potassium, calcium, zinc or magnesium.
  • the pharmaceutical salts of the present invention can be synthesized from the parent compound which contains a basic or acidic moiety by conventional chemical methods.
  • such salts can be prepared via reaction the free acidic or basic forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent, or in a mixture of the two.
  • non-aqueous media like ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred. Lists of suitable salts are found in Remington’s Pharmaceutical Sciences, 17 th ed., Mack Publishing Company, Easton, PA, 1985, p. 1418, the disclosure of which is hereby incorporated by reference.
  • administering refers to any mode of transferring, delivering, introducing or transporting a pharmaceutical drug or other agent to a subject. Such modes include oral administration, topical contact, intravenous, intraperitoneal, intramuscular, intralesional, intranasal, subcutaneous or intrathecal administration. Also contemplated by the present invention is utilization of a device or instrument in administering an agent. Such device may utilize active or passive transport and may be slow-release or fast-release delivery device.
  • ACES N- (2-Acetamido) -2-aminoethanesulfonic acid
  • ADA N- (2-Acetamido) iminodiacetic acid, N- (Carbamoylmethyl) iminodiacetic acid
  • AMPD (2-amino-2-methyl-1, 3-propanediol) ) is a useful buffer at pH 7.8 -9.7.
  • Bicine N, N-Bis (2-hydroxyethyl) glycine
  • BisTris propane (1, 3-Bis [tris (hydroxymethyl) methylamino] propane) .
  • DIPSO N, N-Bis (2-hydroxyethyl) -3-amino-2-hydroxypropanesulfonic acid
  • HEBPS N- (2-Hydroxyethyl) piperazine-N′- (4-butanesulfonic acid)
  • HEPES EPPS N- (2-Hydroxyethyl) piperazine-N′- (4-butanesulfonic acid
  • HEPES (4- (2-hydroxyethyl) -1-piperazineethanesulfonic acid ; 2-morpholinoethanesulfonic acid; 2- (4-morpholino) ethanesulphonic acid; 2- (N-morpholino) ethanesulfonic acid; morpholine-4-thanesulfonic acid hydrate) is widely used to buffer at pH 6.8 -8.2; pKa at 20°C: 7.45-7.65)
  • HEPPSO (2-Hydroxyethyl) piperazine-1- (2-hydroxypropanesulfonic acid) hydrate
  • MES (2- (N-morpholino) ethanesulfonic acid, monohydrate) is used as buffering agent at pH 5.2-7.1 (pKa: 6.16) .
  • MOBS (4-Morpholinebutanesulfonic acid; 3- (N-Morpholino) butanesulfonic acid hemisodium salt) is an homolog of MES and MOPS with higher pKa/It is used to buffer solution at pH6.9-8.3 (pKa: 7.6) .
  • MOPS (4-Morpholinepropanesulfonic acid Sodium salt) .
  • MOPSO ⁇ -Hydroxy-4-morpholinepropanesulfonic acid, 3-Morpholino-2-hydroxypropanesulfonic acid
  • POPSO Piperazine-1, 4-bis (2-hydroxypropanesulfonic acid) dihydrate
  • TAPS [ (2-Hydroxy-1, 1-bis (hydroxymethyl) ethyl) amino] -1-propanesulfonic acid
  • TAPSO (2-Hydroxy-3- [tris (hydroxymethyl) methylamino] -1-propanesulfonic acid) .
  • Tricine (Piperazine-N, N'-Bis [2-Hydroxypropanesulfonic Acid) ] is used to buffer at pH7.4-8.8 (pKa: 8.16) .
  • antibody is used herein in the broadest sense and encompasses various antibody structures, including but not limited to monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies) , and antibody fragments so long as they exhibit the desired antigen-binding activity and fusion proteins comprising an antibody, and any other modified configuration of the immunoglobulin molecule that comprises an antigen recognition site.
  • An antibody includes an antibody of any class, such as IgG, IgA, or IgM (or sub-class thereof) , and the antibody need not be of any particular class.
  • immunoglobulins can be assigned to different classes.
  • immunoglobulins There are five major classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, and several of these may be further divided into subclasses (isotypes) , e.g., IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2.
  • the heavy-chain constant regions that correspond to the different classes of immunoglobulins are called alpha, delta, epsilon, gamma, and mu, respectively.
  • the subunit structures and three-dimensional configurations of different classes of immunoglobulins are well known.
  • antibody fragment refers to a molecule other than an intact antibody that comprises a portion of an intact antibody and that binds the antigen to which the intact antibody binds.
  • antibody fragments include but are not limited to Fv, Fab, Fab', Fab'-SH, F (ab') 2; diabodies; linear antibodies; single-chain antibody molecules (e.g. scFv) ; and multispecific antibodies formed from antibody fragments.
  • a “humanized” antibody refers to a chimeric antibody comprising amino acid residues from non-human HVRs and amino acid residues from human FRs.
  • a humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the HVRs (e.g., CDRs) correspond to those of a non-human antibody, and all or substantially all of the FRs correspond to those of a human antibody.
  • a humanized antibody optionally may comprise at least a portion of an antibody constant region derived from a human antibody.
  • a “humanized form” of an antibody, e.g., a non-human antibody refers to an antibody that has undergone humanization.
  • the term “variable region” or “variable domain” refers to the domain of an antibody heavy or light chain that is involved in binding the antibody to antigen.
  • variable domains of the heavy chain and light chain (VH and VL, respectively) of a native antibody generally have similar structures, with each domain comprising four conserved framework regions (FRs) and three hypervariable regions (HVRs) .
  • FRs conserved framework regions
  • HVRs hypervariable regions
  • a single VH or VL domain may be sufficient to confer antigen-binding specificity.
  • antibodies that bind a particular antigen may be isolated using a VH or VL domain from an antibody that binds the antigen to screen a library of complementary VL or VH domains, respectively. See, e.g., Portolano et al., J. Immunol. 150: 880-887 (1993) ; Clarkson et al., Nature 352: 624-628 (1991) .
  • “monoclonal antibody” refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical except for possible naturally-occurring mutations that may be present in minor amounts. Monoclonal antibodies are highly specific, being directed against a single antigenic site. Furthermore, in contrast to polyclonal antibody preparations, which typically include different antibodies directed against different determinants (epitopes) , each monoclonal antibody is directed against a single determinant on the antigen.
  • the modifier “monoclonal” indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies, and is not to be construed as requiring production of the antibody by any particular method.
  • the monoclonal antibodies to be used in accordance with the present invention may be made by the hybridoma method first described by Kohler and Milstein, Nature 256: 495, 1975, or may be made by recombinant DNA methods such as described in U.S. Pat. No. 4,816,567.
  • the monoclonal antibodies may also be isolated from phage libraries generated using the techniques described in McCafferty et al., Nature 348: 552-554, 1990, for example.
  • humanized antibody refers to forms of non-human (e.g. murine) antibodies that are chimeric immunoglobulins, immunoglobulin chains, or fragments thereof (such as Fv, Fab, Fab', F (ab') 2 or other antigen binding subsequences of antibodies) that contain minimal sequence derived from non-human immunoglobulin.
  • humanized antibodies are human immunoglobulins (recipient antibody) in which residues from a complementarity determining region (CDR) of the recipient are replaced by residues from a CDR of a non-human species (donor antibody) such as mouse, rat, or rabbit having the desired specificity, affinity, and capacity.
  • CDR complementarity determining region
  • Fv framework region (FR) residues of the human immunoglobulin are replaced by corresponding non-human residues.
  • the humanized antibody may comprise residues that are found neither in the recipient antibody nor in the imported CDR or framework sequences, but are included to further refine and optimize antibody performance.
  • the humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the CDR regions correspond to those of a non-human immunoglobulin and all or substantially all of the FR regions are those of a human immunoglobulin consensus sequence.
  • the humanized antibody optimally also will comprise at least a portion of an immunoglobulin constant region or domain (Fc) , typically that of a human immunoglobulin.
  • CDR L1, CDR L2, CDR L3, CDR H1, CDR H2, or CDR H3 are altered with respect to the original antibody, which are also termed one or more CDRs “derived from” one or more CDRs from the original antibody.
  • human antibody means an antibody having an amino acid sequence corresponding to that of an antibody produced by a human and/or which has been made using any of the techniques for making human antibodies known to those skilled in the art or disclosed herein.
  • This definition of a human antibody includes antibodies comprising at least one human heavy chain polypeptide or at least one human light chain polypeptide.
  • One such example is an antibody comprising murine light chain and human heavy chain polypeptides.
  • Human antibodies can be produced using various techniques known in the art. In one embodiment, the human antibody is selected from a phage library, where that phage library expresses human antibodies (Vaughan et al., Nature Biotechnology, 14: 309-314, 1996; Sheets et al., Proc. Natl. Acad.
  • Human antibodies can also be made by immunization of animals into which human immunoglobulin loci have been transgenically introduced in place of the endogenous loci, e.g., mice in which the endogenous immunoglobulin genes have been partially or completely inactivated. This approach is described in U.S. Pat. Nos. 5,545,807; 5,545,806; 5,569,825; 5,625,126; 5,633,425; and 5,661,016.
  • the human antibody may be prepared by immortalizing human B lymphocytes that produce an antibody directed against a target antigen (such B lymphocytes may be recovered from an individual or from single cell cloning of the cDNA, or may have been immunized in vitro) . See, e.g., Cole et al. Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, p. 77, 1985; Boerner et al., J. Immunol., 147 (1) : 86-95, 1991; and U.S. Pat. No. 5,750,373.
  • chimeric antibody is intended to refer to antibodies in which the variable region sequences are derived from one species and the constant region sequences are derived from another species, such as an antibody in which the variable region sequences are derived from a mouse antibody and the constant region sequences are derived from a human antibody.
  • polypeptide oligopeptide
  • peptide peptide and protein are used interchangeably herein to refer to chains of amino acids of any length, preferably, relatively short (e.g., 10-100 amino acids) .
  • the chain may be linear or branched, it may comprise modified amino acids, and/or may be interrupted by non-amino acids.
  • the terms also encompass an amino acid chain that has been modified naturally or by intervention; for example, disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, or any other manipulation or modification, such as conjugation with a labeling component.
  • polypeptides containing one or more analogs of an amino acid including, for example, unnatural amino acids, etc.
  • polypeptides can occur as single chains or associated chains.
  • a “monovalent antibody” comprises one antigen binding site per molecule (e.g., IgG or Fab) .
  • a monovalent antibody can have more than one antigen binding sites, but the binding sites are from different antigens.
  • a “monospecific antibody” comprises two identical antigen binding sites per molecule (e.g. IgG) such that the two binding sites bind identical epitope on the antigen. Thus, they compete with each other on binding to one antigen molecule. Most antibodies found in nature are monospecific. In some instances, a monospecific antibody can also be a monovalent antibody (e.g. Fab) .
  • bivalent antibody comprises two antigen binding sites per molecule (e.g., IgG) . In some instances, the two binding sites have the same antigen specificities. However, bivalent antibodies may be bispecific.
  • bispecific or dual-specific is a hybrid antibody having two different antigen binding sites.
  • the two antigen binding sites of a bispecific antibody bind to two different epitopes, which may reside on the same or different protein targets.
  • a “bifunctional” is antibody is an antibody having identical antigen binding sites (i.e., identical amino acid sequences) in the two arms but each binding site can recognize two different antigens.
  • heteromultimer is a molecule comprising at least a first polypeptide and a second polypeptide, wherein the second polypeptide differs in amino acid sequence from the first polypeptide by at least one amino acid residue.
  • the heteromultimer can comprise a “heterodimer” formed by the first and second polypeptide or can form higher order tertiary structures where polypeptides in addition to the first and second polypeptide are present.
  • heterodimer is a molecule comprising a first polypeptide and a second polypeptide, wherein the second polypeptide differs in amino acid sequence from the first polypeptide by at least one amino acid residue.
  • the “hinge region” includes the meaning known in the art, which is illustrated in, for example, Janeway et al., ImmunoBiology: the immune system in health and disease, (Elsevier Science Ltd., NY) (4th ed., 1999) ; Bloom et al., Protein Science (1997) , 6: 407-415; Humphreys et al., J. Immunol. Methods (1997) , 209: 193-202.
  • immunoglobulin-like hinge region refers to the hinge region and hinge sequence of an immunoglobulin-like or an antibody-like molecule (e.g., immunoadhesins) .
  • the immunoglobulin-like hinge region can be from or derived from any IgG1, IgG2, IgG3, or IgG4 subtype, or from IgA, IgE, IgD or IgM, including chimeric forms thereof, e.g., a chimeric IgG1/2 hinge region.
  • immune effector cell refers to a cell within the natural repertoire of cells in the human immune system which can be activated to affect the viability of a target cell.
  • the viability of a target cell can include cell survival, proliferation, and/or ability to interact with other cells.
  • Antibodies of the invention can be produced using techniques well known in the art, e.g., recombinant technologies, phage display technologies, synthetic technologies or combinations of such technologies or other technologies readily known in the art (see, for example, Jayasena, S. D., Clin. Chem., 45: 1628-50, 1999 and Fellouse, F. A., et al, J. Mol. Biol., 373 (4) : 924-40, 2007) .
  • cytotoxic agent refers to a substance that inhibits or prevents a cellular function and/or causes cell death or destruction. Cytotoxic agents include, but are not limited to, radioactive isotopes (e.g., At211, I131, I125, Y90, In111, Re186, Re188, Sm153, Bi212, P32, Pb212, Zr89, F18, and radioactive isotopes of Lu, e.g.
  • chemotherapeutic agents or drugs e.g., tubulysin, maytansin, auristatin, DNA minor groove binders (such as PBD dimers) , ducarmysin, topoisomerase inhibitor, RNA polymerase inhibitors, DNA alkylators, methotrexate, adriamicin, vinca alkaloids (vincristine, vinblastine, etoposide) , doxorubicin, melphalan, mitomycin C, chlorambucil, daunorubicin or other intercalating agents) ; growth inhibitory agents; enzymes and fragments thereof such as nucleolytic enzymes; antibiotics; toxins such as small molecule toxins or enzymatically active toxins of bacterial, fungal, plant or animal origin, including fragments and/or variants thereof; and the various antitumor or anticancer agents disclosed throughout the application.
  • Linker refers to a chemical moiety comprising a covalent bond or a chain of atoms that covalently attaches an antibody to a drug moiety.
  • linkers include a divalent radical such as an alkyldiyl, an aryldiyl, a heteroaryldiyl, moieties such as: -- (CR2) nO (CR2) n--, repeating units of alkyloxy (e.g. polyethylenoxy, PEG, polymethyleneoxy) and alkylamino (e.g. polyethyleneamino) ; and diacid ester and amides including succinate, succinamide, diglycolate, malonate, and caproamide.
  • linkers can comprise one or more amino acid residues, such as valine, phenylalanine, lysine, and homolysine.
  • the key factor of the invention in the conjugation process is the transition metal cation-amino complex, M (NR 1 R 2 R 3 ) m1 m2+ , which coordinate the selective reduction of certain chain of disulfide bonds of biomolecule.
  • M transition metal cation-amino complex
  • the certain inter-chain disulfide bonds in an antibody Preferably the certain inter-chain disulfide bonds in an antibody.
  • disulfide bonds in the CH2 domain were the most susceptible to reduction.
  • Disulfide bonds in VL, CL, VH, and CH1 domains had similar and moderate susceptibility, while disulfide bonds in the CH3 domain were the least susceptible to reduction (Liu, H, et al Anal. Chem., 2010, 82, 5219–5226) .
  • the using of ZnCl 2 salt at low temperatures of 2 ⁇ 8 °C in coordination the reduction of the disulfide bonds of a IgG antibody of the invention WO2020164561 made practically possible of the distinguishable reduction above.
  • the transition metal cation-amino complex, M (NR 1 R 2 R 3 ) m1 m2+ which is used in the conjugation process of the invention, is much bulky, not only can coordinate the disulfide reduction, but also stereoscopically hinders the reductant (such as TCEP) to access to disulfide bonds between the two heavy chains of an IgG antibody, thus results in much better selective reduction and following by conjugation with a drug/linker complex.
  • (NR 1 R 2 R 3 ) m1 can form a dimer, trimer, tetramer, pentamer, or hexamer wherein these polymers are covalently linked among N, R 1 , R 2 and R 3 ; and N, R 1 , R 2 and/or R 3 themselve can form heterocyclic, carbocyclic, diheterocyclic, or dicarbocyclic rings.
  • the preferred M is Zn
  • the preferred M (NR 1 R 2 R 3 ) m1 m2+ are exampled as following: Zn (NH 2 CH 3 ) 2 2+ , Zn (NH 2 CH 2 CH 3 ) 2 2+ , Zn (NH 2 CH 2 CH 2 CH 3 ) 2 2+ , Zn (NH 2 CH (CH 3 ) 2 ) 2 2+ , Zn (NH 2 C (CH 3 ) 3 ) 2 2+ , Zn (NH 2 CH 2 C (CH 3 ) 3 ) 2 2+ , Zn (NH (CH 3 ) 2 ) 2 2+ , Zn (NH (CH 2 CH 3 ) 2 ) 2 2+ , Zn (NH (CH (CH 3 ) 2 ) 2 ) 2 2+ , Zn (NH (CH (CH 3 ) 2 ) 2 ) 2 2+ , Zn (NH (CH (CH 3 ) 2 ) 2 ) 2+ , Zn (NH (C (CH 3 ) 3
  • All the complex cations above can be formed with an anion, selected from, but not limited, Cl - , Br - , I - , SO 4 2- , HSO 4 - , NO 3 - , PO 4 3- , HPO 4 2- , H 2 PO 4 - , CO 3 2- , HCO 3 - , HCOO - , CH 3 COO - , F 3 CCOO - , Cl 3 CCOO - , FCH 2 COO - , ClCH 2 COO - , F 2 CHCOO - , Cl 2 CHCOO - , BF 4 - , SO 3 2- , HSO 3 - , CH 3 SO 3- , C 6 H 5 CH 2 SO 3- , C 6 H 5 SO 3- , C 6 H 5 COO - , C 6 H 5 CH 2 COO - , C 6 F 5 O - , C 6 H 4 (OH) COO - , C 6 H 2 F 3 O - , C
  • the transition metal cation-amino complex in the reaction solution are 0.5 ⁇ 20 equivalents of the antibody, preferably 1.0 -5.0 equivalents of the antibody, more preferably 1.5 -3.0 equivalents of the antibody.
  • the transition metal cation-amino complex can be added to the reaction solution with a water-miscible organic solvent, selected from, but not limited, ethanol, methanol, propanol, propandiol, DMA, DMF, DMSO, THF, or CH 3 CN.
  • the reductant is selected from TECP or P (CH 2 CH 2 CH 2 OH) 3 , and more preferably the reductant is selected from TECP.
  • concentration of the reductant in the reaction solution may be 0.04 mM -0.4 mM, or 1.0 -10.0 equivalents of antibody used in the reaction.
  • the reductant is used at 2.0 -4.0 equivalents of an antibody.
  • the optimum buffer for conduction of the selective reduction is selected from, but not limited, PBS, Mes, Bis-Tris, Bis-Tris Propane, Pipes, Aces, Mopso, Bes, Mops, Hepes, Tes, Pipps, Dipso, Tapso, Heppso, Tris-up, Tris-HCl, Tricine, Hepps, Gly-Gly, Bicine, Taps, Hepee, Acetates, Histidine, Citrates, MES, Borates, or combinations two, three or four buffer components from above.
  • the pH of the buffer is selected 4.0 -9.0, preferred 5.0 -7.5, more preferred 5.5 -7.5.
  • the concentration of the buffer in the reaction is 0.02 –1.0 M, preferably 20 –200 mM, more preferably 20 –100 mM.
  • concentration of the buffer in the reaction is 0.02 –1.0 M, preferably 20 –200 mM, more preferably 20 –100 mM.
  • up to 30%of water mixable (miscible) organic solvents selected from DMA, DMF, ethanol, methanol, acetone, acetonitrile, THF, isopropanol, dioxane, propylene glycol, or ethylene diol can be added as the co-solvent in water based buffer solution;
  • the optimum temperature for the reduction reaction is typically controlled between about -5 and 40 °C, and the reaction time is 15 minutes to 48 hours. But it is well-understandable in the field of protein conjugation that the reaction time and temperature can be determined by those skilled in the art based on the specific protein, in particular, the antibody to be conjugated.
  • a preferable reduction reaction can be controlled at a temperature typically between about -5 to about 40 °C, and preferably, about 0 to 37 °C; more preferably about 2 to 8 °C, and more procisily 4 ⁇ 1 °C.
  • the process of the conjugation is 15 min to 12 hours, and more preferably at a temperature between about 2 and 8 °C, and the process time is about 30 min to 15 hours (overnight) .
  • a Drug/linker complex/assembly is directly added to the solution of the reduction reaction for conjugation.
  • the Drug/linker complex/assembly having a formula (I) or (II) represented as:
  • Lv 1 and Lv 2 are a thiol reaction group, and are independently selected from:
  • X 1 ’ and X 2 ’ are independently F, Cl, Br, I, OTf, OMs, OC 6 H 4 (NO 2 ) , OC 6 H 3 (NO 2 ) 2 , OC 6 F 5 , OC 6 HF 4 , or Lv 3 ;
  • X 2 is O, NH, N (R 1 ) , or CH 2 ;
  • R 3 and R 5 are independently H, R 1 , aromatic, heteroaromatic, or aromatic group wherein one or several H atoms are replaced independently by -R 1 , -halogen, -OR 1 , -SR 1 , -NR 1 R 2 , -NO 2 , -S (O) R 1 , -S (O) 2 R 1, or -COOR 1 ;
  • Lv 3 and Lv 3 ’ are independently a leaving group selected from F, Cl, Br, I, nitrophenoxyl; N-hydroxysuccinimide (NHS) ; phenoxyl;
  • L 1 and L 2 are, the same or different, independently selected from O, NH, S, NHNH, N (R 3 ) , N (R 3 ) N (R 3’ ) , polyethyleneoxy unit of formula (OCH 2 CH 2 ) p OR 3 , or (OCH 2 CH (CH 3 ) ) p OR 3 , or NH (CH 2 CH 2 O) p R 3 , or NH (CH 2 CH (CH 3 ) O) p R 3 , or N [ (CH 2 CH 2 O) p R 3 ] [ (CH 2 CH 2 O) p’ R 3’ ] , or (OCH 2 CH 2 ) p COOR 3 , or CH 2 CH 2 (OCH 2 CH 2 ) p COOR 3 , wherein p and p’ are independently an integer selected from 0 to about 1000, or combination thereof; C 1 -C 8 of alkyl; C 2 -C 8 of heteroalkyl, alkylcycloalkyl, heterocycloal
  • L 1 or L 2 may contain a self-immolative or a non-self-immolative component, peptidyl units, a hydrazone bond, a disulfide, an ester, an oxime, an amide, or a thioether bond.
  • the self-immolative unit includes, but is not limited to, aromatic compounds that are electronically similar to the para-aminobenzylcarbamoyl (PAB) groups such as 2-aminoimidazol-5-methanol derivatives, heterocyclic PAB analogs, beta-glucuronide, and ortho or para-aminobenzylacetals.
  • PAB para-aminobenzylcarbamoyl
  • the self-immolative linker component has one of the following structures:
  • (*) atom is the point of attachment of additional spacer or releasable linker units, or the cytotoxic agent, and/or the binding molecule (CBA) ;
  • X 1 , Y 1 , Z 2 and Z 3 are independently NH, O, or S;
  • Z 1 is independently H, NH, O or S;
  • v is 0 or 1;
  • the non-self-immolative linker component is one of the following structures:
  • (*) atom is the point of attachment of additional spacer R 1 or releasable linkers, the cytotoxic agents, and/or the binding molecules;
  • X 1 , Y 1 , U 1 , R 1, R 5 , R 5 ’ are defined as above; r is
  • L 1 or L 2 may be composed of one or more linker components of 6-maleimidocaproyl ( “MC” ) , maleimidopropanoyl ( “MP” ) , valine-citrulline ( “val-cit” or “vc” ) , alanine-phenylalanine ( “ala-phe” or “af” ) , p-aminobenzyloxycarbonyl ( “PAB” ) , 4-thiopentanoate ( “SPP” ) , 4- (N-maleimidomethyl) cyclohexane-1 carboxylate ( “MCC” ) , (4-acetyl) amino-benzoate ( “SIAB” ) , 4-thio-butyrate (SPDB) , 4-thio-2-hydroxysulfonyl-butyrate (2-Sulfo-SPDB) , or natural or unnatural peptides having 1 ⁇ 8 natural or unnatural amino acid unites.
  • L 1 or L 2 may be a releasable linker.
  • the term releasable linker refers to a linker that includes at least one bond that can be broken under physiological conditions, such as a pH-labile, acid-labile, base-labile, oxidatively labile, metabolically labile, biochemically labile, or enzyme-labile bond.
  • physiological conditions resulting in bond breaking do not necessarily include a biological or metabolic process, and instead may include a standard chemical reaction, such as a hydrolysis or substitution reaction, for example, an endosome having a lower pH than cytosolic pH, and/or disulfide bond exchange reaction with a intracellular thiol, such as a millimolar range of abundant of glutathione inside the malignant cells.
  • a standard chemical reaction such as a hydrolysis or substitution reaction, for example, an endosome having a lower pH than cytosolic pH, and/or disulfide bond exchange reaction with a intracellular thiol, such as a millimolar range of abundant of glutathione inside the malignant cells.
  • releasable linkers examples include, but not limited:
  • Example structures of the components of the linker L 1 and L 2 may contain:
  • X 2 , X 3 , X 4 , X 5 , or X 6 are independently selected from NH; NHNH; N (R 12 ) ; N (R 12 ) N (R 12’ ) ; O; S; C 1 -C 6 of alkyl; C 2 -C 6 of heteroalkyl, alkylcycloalkyl, heterocycloalkyl; C 3 -C 8 of aryl, Ar-alkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, heteroaryl; CH 2 OR 12 , CH 2 SR 12 , CH 2 NHR 12 , or 1 ⁇ 8 amino acids; wherein R 12 and R 12’ are independently H; C 1 -C 8 of alkyl; C 2 -C 8 of hetero-alkyl, alkylcycloalkyl, heterocycloalkyl; C 3
  • L 1 , L 2 , X 1 , X 2 , X 3 , X 1’ , X 2’ and X 3’ can be independently absent.
  • E 1 is a joint group that link two thiol reactonable groups of Lv 1 and Lv 2 .
  • D is a cytotoxic drug, or a therapeutic drug, or an immunotherapeutical protein, or a function molecule for enhancement of binding or stabilization of the cell-binding protein agent, or a cell-surface receptor binding lingand, such as an antybody or an antibody fragment, or siRNA or DNA molecule.
  • the cytotoxic drug is selected from, but not limited to:
  • Chemotherapeutic agents a) . Alkylating agents: such as Nitrogen mustards: chlorambucil, chlornaphazine, cyclophosphamide, dacarbazine, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, mannomustine, mitobronitol, melphalan, mitolactol, pipobroman, novembichin, phenesterine, prednimustine, thiotepa, trofosfamide, uracil mustard; CC-1065 (including its adozelesin, carzelesin and bizelesin synthetic analogues) ; Duocarmycin (including the synthetic analogues, KW-2189 and CBI-TMI) ; Benzodiazepine dimers (e.g., dimmers of pyrrolobenzodiazepine (PBD) or tomaymycin, indolinobenzodiazepine
  • Plant Alkaloids such as Vinca alkaloids: (vincristine, vinblastine, vindesine, vinorelbine, navelbin) ; Taxoids: (paclitaxel, docetaxol) and their analogs, Maytansinoids (DM1, DM2, DM3, DM4, maytansine and ansamitocins) and their analogs, cryptophycins (particularly cryptophycin 1 and cryptophycin 8) ; epothilones, eleutherobin, discodermo-lide, bryostatins, dolostatins, auristatins, tubulysins, cephalostatins; pancratistatin; a sarcodictyin; spongistatin; c) .
  • DNA Topoisomerase Inhibitors such as [Epipodophyllins: (9-aminocamptothecin, camptothecin, crisnatol, daunomycin, etoposide, etoposide phosphate, irinotecan, mitoxantrone, novantrone, retinoic acids (retinols) , teniposide, topotecan, 9-nitrocamptothecin (RFS 2000) ) ; mitomycins: (mitomycin C) ] ; d) .
  • Epipodophyllins (9-aminocamptothecin, camptothecin, crisnatol, daunomycin, etoposide, etoposide phosphate, irinotecan, mitoxantrone, novantrone, retinoic acids (retinols) , teniposide, topotecan, 9-nitrocamptothec
  • Anti-metabolites such as ⁇ [Anti-folate: DHFR inhibitors: (methotrexate, trimetrexate, denopterin, pteropterin, aminopterin (4-aminopteroic acid) or the other folic acid analogues) ; IMP dehydrogenase Inhibitors: (mycophenolic acid, tiazofurin, ribavirin, EICAR) ; Ribonucleotide reductase Inhibitors: (hydroxyurea, deferoxamine) ] ; [Pyrimidine analogs: Uracil analogs: (ancitabine, azacitidine, 6-azauridine, capecitabine (Xeloda) , carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, 5-Fluorouracil, floxuridine, ratitrexed (Tomudex) ) ; Cytosine analogs: (cytar
  • Hormonal therapies such as ⁇ Receptor antagonists: [Anti-estrogen: (megestrol, raloxifene, tamoxifen) ; LHRH agonists: (goscrclin, leuprolide acetate) ; Anti-androgens: (bicalutamide, flutamide, calusterone, dromostanolone propionate, epitiostanol, goserelin, leuprolide, mepitiostane, nilutamide, testolactone, trilostane and other androgens inhibitors) ] ; Retinoids/Deltoids: [Vitamin D3 analogs: (CB 1093, EB 1089 KH 1060, cholecalciferol, ergocalciferol) ; Photodynamic therapies: (verteporfin, phthalocyanine, photosensitizer Pc4, demethoxy-hypocrellin A) ; Cytokines
  • Kinase inhibitors such as BIBW 2992 (anti-EGFR/Erb2) , imatinib, gefitinib, pegaptanib, sorafenib, dasatinib, sunitinib, erlotinib, nilotinib, lapatinib, axitinib, pazopanib.
  • vandetanib E7080 (anti-VEGFR2) , mubritinib, ponatinib (AP24534) , bafetinib (INNO-406) , bosutinib (SKI-606) , cabozantinib, vismodegib, iniparib, ruxolitinib, CYT387, axitinib, tivozanib, sorafenib, bevacizumab, cetuximab, Trastuzumab, Ranibizumab, Panitumumab, ispinesib; g) . antibiotics, such as the enediyne antibiotics (e.g.
  • calicheamicins especially calicheamicin ⁇ 1, ⁇ 1, ⁇ 1 and ⁇ 1, see, e.g., J. Med. Chem., 39 (11) , 2103–2117 (1996) , Angew Chem Intl. Ed. Engl.
  • dynemicin including dynemicin A and deoxydynemicin
  • esperamicin including dynemicin A and deoxydynemicin
  • esperamicin including dynemicin A and deoxydynemicin
  • esperamicin including dynemicin A and deoxydynemicin
  • esperamicin including dynemicin A and deoxydynemicin
  • esperamicin including kedarcidin, C-1027, maduropeptin
  • neocarzinostatin chromophore and related chromoprotein enediyne antiobiotic chromomophores , aclacinomysins, actinomycin, authramycin, azaserine, bleomycins, cactinomycin, carabicin, carminomycin, carzinophilin
  • chromomycins dactinomycin, daun
  • acetogenins especially bullatacin and bullatacinone
  • gemcitabine epoxomicins (e.g. carfilzomib) , bortezomib, thalidomide, lenalidomide, pomalidomide, tosedostat, zybrestat, PLX4032, STA-9090, Stimuvax, allovectin-7, Xegeva, Provenge, Yervoy, Isoprenylation inhibitors (such as Lovastatin) , Dopaminergic neurotoxins (such as 1-methyl-4-phenylpyridinium ion) , Cell cycle inhibitors (such as staurosporine) , Actinomycins (such as Actinomycin D, dactinomycin) , Bleomycins (such as bleomycin A2, bleomycin B2, peplomycin) , Anthracyclines (such as daunorubi
  • An anti-autoimmune disease agent includes, but is not limited to, cyclosporine, cyclosporine A, aminocaproic acid, azathioprine, bromocriptine, chlorambucil, chloroquine, cyclophosphamide, corticosteroids (e.g.
  • amcinonide betamethasone, budesonide, hydrocortisone, flunisolide, fluticasone propionate, fluocortolone danazol, dexamethasone, Triamcinolone acetonide, beclometasone dipropionate) , DHEA, enanercept, hydroxychloroquine, infliximab, meloxicam, methotrexate, mofetil, mycophenylate, prednisone, sirolimus, tacrolimus.
  • An anti-infectious disease agent includes, but is not limited to, a) .
  • Aminoglycosides amikacin, astromicin, gentamicin (netilmicin, sisomicin, isepamicin) , hygromycin B, kanamycin (amikacin, arbekacin, bekanamycin, dibekacin, tobramycin) , neomycin (framycetin, paromomycin, ribostamycin) , netilmicin, spectinomycin, streptomycin, tobramycin, verdamicin; b) .
  • Amphenicols azidamfenicol, chloramphenicol, florfenicol, thiamphenicol; c) .
  • Ansamycins geldanamycin, herbimycin; d) .
  • Carbapenems biapenem, doripenem, ertapenem, imipenem/cilastatin, meropenem, panipenem; e) .
  • Cephems carbacephem (loracarbef) , cefacetrile, cefaclor, cefradine, cefadroxil, cefalonium, cefaloridine, cefalotin or cefalothin, cefalexin, cefaloglycin, cefamandole, cefapirin, cefatrizine, cefazaflur, cefazedone, cefazolin, cefbuperazone, cefcapene, cefdaloxime, cefepime, cefminox, cefoxitin, cefprozil, cefroxadine, ceftezole, cefuroxime, cefixime, cefdinir, cefditoren, cefepime, cefetamet, cefmenoxime, cefodizime, cefonicid, cefoperazone, ceforanide, cefotaxime, cefotiam, cefozopran, cephal
  • Glycopeptides bleomycin, vancomycin (oritavancin, telavancin) , teicoplanin (dalbavancin) , ramoplanin; g) .
  • Glycylcyclines e.g. tigecycline; g) .
  • ⁇ -Lactamase inhibitors penam (sulbactam, tazobactam) , clavam (clavulanic acid) ; i) .
  • Lincosamides clindamycin, lincomycin; j) .
  • Lipopeptides daptomycin, A54145, calcium-dependent antibiotics (CDA) ; k) .
  • Macrolides azithromycin, cethromycin, clarithromycin, dirithromycin, erythromycin, flurithromycin, josamycin, ketolide (telithromycin, cethromycin) , midecamycin, miocamycin, oleandomycin, rifamycins (rifampicin, rifampin, rifabutin, rifapentine) , rokitamycin, roxithromycin, spectinomycin, spiramycin, tacrolimus (FK506) , troleandomycin, telithromycin; l) .
  • Monobactams aztreonam, tigemonam; m) .
  • Oxazolidinones linezolid; n) .
  • Penicillins amoxicillin, ampicillin (pivampicillin, hetacillin, bacampicillin, metampicillin, talampicillin) , azidocillin, azlocillin, benzylpenicillin, benzathine benzylpenicillin, benzathine phenoxymethyl-penicillin, clometocillin, procaine benzylpenicillin, carbenicillin (carindacillin) , cloxacillin, dicloxacillin, epicillin, flucloxacillin, mecillinam (pivmecillinam) , mezlocillin, meticillin, nafcillin, oxacillin, penamecillin, penicillin, pheneticillin, phenoxymethylpenicillin, piperacillin, propicillin, sulbenicillin, temocillin, ticarcillin; o) .
  • Polypeptides bacitracin, colistin, polymyxin B; p) .
  • Quinolones alatrofloxacin, balofloxacin, ciprofloxacin, clinafloxacin, danofloxacin, difloxacin, enoxacin, enrofloxacin, floxin, garenoxacin, gatifloxacin, gemifloxacin, grepafloxacin, kano trovafloxacin, levofloxacin, lomefloxacin, marbofloxacin, moxifloxacin, nadifloxacin, norfloxacin, orbifloxacin, ofloxacin, pefloxacin, trovafloxacin, grepafloxacin, sitafloxacin, sparfloxacin, temafloxacin, tosufloxacin, trovafloxacin; q) .
  • Streptogramins pristinamycin, quinupristin/dalfopristin) ; r) .
  • Sulfonamides mafenide, prontosil, sulfacetamide, sulfamethizole, sulfanilimide, sulfasalazine, sulfisoxazole, trimethoprim, trimethoprim-sulfamethoxazole (co-trimoxazole) ; s) .
  • Steroid antibacterials e.g. fusidic acid; t) .
  • Tetracyclines doxycycline, chlortetracycline, clomocycline, demeclocycline, lymecycline, meclocycline, metacycline, minocycline, oxytetracycline, penimepicycline, rolitetracycline, tetracycline, glycylcyclines (e.g. tigecycline) ; u) .
  • antibiotics include annonacin, arsphenamine, bactoprenol inhibitors (Bacitracin) , DADAL/AR inhibitors (cycloserine) , dictyostatin, discodermolide, eleutherobin, epothilone, ethambutol, etoposide, faropenem, fusidic acid, furazolidone, isoniazid, laulimalide, metronidazole, mupirocin, mycolactone, NAM synthesis inhibitors (e.g.
  • fosfomycin nitrofurantoin, paclitaxel, platensimycin, pyrazinamide, quinupristin/dalfopristin, rifampicin (rifampin) , tazobactam tinidazole, uvaricin;
  • Anti-viral drugs a) . Entry/fusion inhibitors: aplaviroc, maraviroc, vicriviroc, gp41 (enfuvirtide) , PRO 140, CD4 (ibalizumab) ; b) . Integrase inhibitors: raltegravir, elvitegravir, globoidnan A; c) . Maturation inhibitors: bevirimat, becon; d) . Neuraminidase inhibitors: oseltamivir, zanamivir, peramivir; e) .
  • Nucleosides &nucleotides abacavir, aciclovir, adefovir, amdoxovir, apricitabine, brivudine, cidofovir, clevudine, dexelvucitabine, didanosine (ddI) , elvucitabine, emtricitabine (FTC) , entecavir, famciclovir, fluorouracil (5-FU) , 3’-fluoro-substituted 2’, 3’-dideoxynucleoside analogues (e.g.
  • ⁇ -l-thymidine and ⁇ -l-2’-deoxycytidine penciclovir, racivir, ribavirin, stampidine, stavudine (d4T) , taribavirin (viramidine) , telbivudine, tenofovir, trifluridine valaciclovir, valganciclovir, zalcitabine (ddC) , zidovudine (AZT) ; f) .
  • Non-nucleosides amantadine, ateviridine, capravirine, diarylpyrimidines (etravirine, rilpivirine) , delavirdine, docosanol, emivirine, efavirenz, foscarnet (phosphonoformic acid) , imiquimod, interferon alfa, loviride, lodenosine, methisazone, nevirapine, NOV-205, peginterferon alfa, podophyllotoxin, rifampicin, rimantadine, resiquimod (R-848) , tromantadine; g) .
  • Protease inhibitors amprenavir, atazanavir, boceprevir, darunavir, fosamprenavir, indinavir, lopinavir, nelfinavir, pleconaril, ritonavir, saquinavir, telaprevir (VX-950) , tipranavir; h) .
  • anti-virus drugs abzyme, arbidol, calanolide a, ceragenin, cyanovirin-n, diarylpyrimidines, epigallocatechin gallate (EGCG) , foscarnet, griffithsin, taribavirin (viramidine) , hydroxyurea, KP-1461, miltefosine, pleconaril, portmanteau inhibitors, ribavirin, seliciclib.
  • EGCG epigallocatechin gallate
  • griffithsin taribavirin (viramidine)
  • hydroxyurea KP-1461
  • miltefosine pleconaril
  • portmanteau inhibitors ribavirin, seliciclib.
  • the drugs used for conjugates via a bridge linker of the present invention also include radioisotopes.
  • radioisotopes are 3 H, 11 C, 14 C, 18 F, 32 P, 35 S, 64 Cu, 68 Ga, 86 Y, 99 Tc, 111 In, 123 I, 124 I, 125 I, 131 I, 133 Xe, 177 Lu, 211 At, or 213 Bi.
  • Radioisotope labeled antibodies are useful in receptor targeted imaging experiments or can be for targeted treatment such as with the antibody-drug conjugates of the invention (Wu et al (2005) Nature Biotechnology 23 (9) : 1137-46) .
  • the cell binding molecules e.g.
  • an antibody can be labeled with ligand reagents through the bridge linkers of the present patent that bind, chelate or otherwise complex a radioisotope metal, using the techniques described in Current Protocols in Immunology, Volumes 1 and 2, Coligen et al, Ed. Wiley-Interscience, New York, Pubs. (1991) .
  • Chelating ligands which may complex a metal ion include DOTA, DOTP, DOTMA, DTPA and TETA (Macrocyclics, Dallas, Tex. USA) .
  • the drug D can be a chromophore molecule, for which the conjugate can be used for detection, monitoring, or study the interaction of the cell binding molecule with a target cell.
  • Chromophore molecules are a compound that have the ability to absorb a kind of light, such as UV light, florescent light, IR light, near IR light, visual light;
  • a chromatophore molecule includes a class or subclass of xanthophores, erythrophores, iridophores, leucophores, melanophores, and cyanophores; a class or subclass of fluorophore molecules which are fluorescent chemical compounds re-emitting light upon light; a class or subclass of visual phototransduction molecules; a class or subclass of photophore molecules; a class or subclass of luminescence molecules; and a class or subclass of luciferin compounds.
  • the chromophore molecule can be selected from, but not limited, non-protein organic fluorophores, such as: Xanthene derivatives (fluorescein, rhodamine, Oregon green, eosin, and Texas red) ; Cyanine derivatives: (cyanine, indocarbocyanine, oxacarbocyanine, thiacarbocyanine, and merocyanine) ; Squaraine derivatives and ring-substituted squaraines, including Seta, SeTau, and Square dyes; Naphthalene derivatives (dansyl and prodan derivatives) ; Coumarin derivatives; Oxadiazole derivatives (pyridyloxazole, nitrobenzoxadiazole and benzoxadiazole) ; Anthracene derivatives (anthraquinones, including DRAQ5, DRAQ7 and CyTRAK Orange) ; Pyrene derivatives (cascade blue, etc) ; Oxazine derivative
  • Acridine derivatives proflavin, acridine orange, acridine yellow etc.
  • Arylmethine derivatives auramine, crystal violet, malachite green
  • Tetrapyrrole derivatives porphin, phthalocyanine, bilirubin
  • a chromophore molecule can be selected from any analogs and derivatives of the following fluorophore compounds: CF dye (Biotium) , DRAQ and CyTRAK probes (BioStatus) , BODIPY (Invitrogen) , Alexa Fluor (Invitrogen) , DyLight Fluor (Thermo Scientific, Pierce) , Atto and Tracy (Sigma Aldrich) , FluoProbes (Interchim) , Abberior Dyes (Abberior) , DY and MegaStokes Dyes (Dyomics) , Sulfo Cy dyes (Cyandye) , HiLyte Fluor (AnaSpec) , Seta, SeTau and Square Dyes (SETA BioMedicals) , Quasar and Cal Fluor dyes (Biosearch Technologies) , SureLight Dyes (APC, RPEPerCP, Phycobilisomes) (Columbia Biosciences) , A
  • Examples of the widely used fluorophore compounds which are reactive or conjugatable with the linkers of the invention are: Allophycocyanin (APC) , Aminocoumarin, APC-Cy7 conjugates, BODIPY-FL, Cascade Blue, Cy2, Cy3, Cy3.5, Cy3B, Cy5, Cy5.5, Cy7, Fluorescein, FluorX, Hydroxycoumarin, IR-783, Lissamine Rhodamine B, Lucifer yellow, Methoxycoumarin, NBD, Pacific Blue, Pacific Orange, PE-Cy5 conjugates, PE-Cy7 conjugates, PerCP, R-Phycoerythrin (PE) , Red 613, Seta-555-Azide, Seta-555-DBCO, Seta-555-NHS, Seta-580-NHS, Seta-680-NHS, Seta-780-NHS, Seta-APC-780, Seta-PerCP-680, Seta-R-PE-670, SeTau-380-NHS, SeT
  • the fluorophore compounds that can be linked to the linkers of the invention for study of nucleic acids or proteins are selected from the following compounds or their derivatives: 7-AAD (7-aminoactinomycin D, CG-selective) , Acridine Orange, Chromomycin A3, CyTRAK Orange (Biostatus, red excitation dark) , DAPI, DRAQ5, DRAQ7, Ethidium Bromide, Hoechst33258, Hoechst33342, LDS 751, Mithramycin, PropidiumIodide (PI) , SYTOX Blue, SYTOX Green, SYTOX Orange, Thiazole Orange, TO-PRO: Cyanine Monomer, TOTO-1, TO-PRO-1, TOTO-3, TO-PRO-3, YOSeta-1, YOYO-1.
  • 7-AAD 7-aminoactinomycin D, CG-selective
  • Acridine Orange Chromomycin A3, CyTRAK Orange (Bio
  • the fluorophore compounds that can be linked to the linkers of the invention for study cells are selected from the following compounds or their derivatives: DCFH (2'7'Dichorodihydro-fluorescein, oxidized form) , DHR (Dihydrorhodamine 123, oxidized form, light catalyzes oxidation) , Fluo-3 (AM ester. pH > 6) , Fluo-4 (AM ester. pH 7.2) , Indo-1 (AM ester, low/high calcium (Ca2+) ) , and SNARF (pH 6/9) .
  • the preferred fluorophore compounds that can be linked to the linkers of the invention for study proteins/antibodies are selected from the following compounds or their derivatives: Allophycocyanin (APC) , AmCyan1 (tetramer, Clontech) , AsRed2 (tetramer, Clontech) , Azami Green (monomer, MBL) , Azurite, B-phycoerythrin (BPE) , Cerulean, CyPet, DsRed monomer (Clontech) , DsRed2 ( “RFP” , Clontech) , EBFP, EBFP2, ECFP, EGFP (weak dimer, Clontech) , Emerald (weak dimer, Invitrogen) , EYFP (weak dimer, Clontech) , GFP (S65A mutation) , GFP (S65C mutation) , GFP (S65L mutation) , GFP (S65T mutation) , GFP (
  • the drug D can be polyalkylene glycols that are used for extending the half-life of the cell-binding antibody, or antibody-like protein molecule when administered to a mammal.
  • Polyalkylene glycols include, but are not limited to, poly (ethylene glycols) (PEGs) , poly (propylene glycol) and copolymers of ethylene oxide and propylene oxide; particularly preferred are PEGs, and more particularly preferred are monofunctionally activated hydroxyPEGs (e.g., hydroxyl PEGs activated at a single terminus, including reactive esters of hydroxyPEG-monocarboxylic acids, hydroxyPEG-monoaldehydes, hydroxyPEG-monoamines, hydroxyPEG-monohydrazides, hydroxyPEG-monocarbazates, hydroxyl PEG-monoiodoacetamides, hydroxyl PEG-monomaleimides, hydroxyl PEG-monoorthopyridyl dis
  • the polyalkylene glycol has a molecular weight of from about 10 Daltons to about 200 kDa, preferably about 88 Da to about 40 kDa; two branches each with a molecular weight of about 88 Da to about 40 kDa; and more preferably two branches, each of about 88 Da to about 20 kDa.
  • the polyalkylene glycol is poly (ethylene) glycol and has a molecular weight of about 10 kDa; about 20 kDa, or about 40 kDa.
  • the PEG is a PEG 10 kDa (linear or branched) , a PEG 20 kDa (linear or branched) , or a PEG 40 kDa (linear or branched) .
  • a number of US patents have disclosed the preparation of linear or branched “non-antigenic” PEG polymers and derivatives or conjugates thereof, e.g., U.S. Pat. Nos.
  • D is more preferably a potent cytotoxic agent, selected from a tubulysin and its analogs, a maytansinoid and its analogs, a taxanoid (taxane) and its analogs, a CC- 1065 and its analogs, a daunorubicin or doxorubicin and its analogs, an amatoxin and its analogs, a benzodiazepine dimer (e.g., dimers of pyrrolobenzodiazepine (PBD) , tomaymycin, anthramycin, indolinobenzodiazepines, imidazobenzothiadiazepines, or oxazolidinobenzo-diazepines) and their analogs, a calicheamicin and the enediyne antibiotic and their analogs, an actinomycin and its analogs, an azaserine and its analogs, a bleomycin and its analogs, an epirubicin and its analogs,
  • Tubulysin and its analogs are well known in the art and can be isolated from natural sources according to known methods or prepared synthetically according to known methods (e.g. Balasubramanian, R., et al. J. Med. Chem., 2009, 52, 238–40; Wipf, P., et al. Org. Lett., 2004, 6, 4057–60; Pando, O., et al. J. Am. Chem. Soc., 2011, 133, 7692–5; Reddy, J.A., et al. Mol. Pharmaceutics, 2009, 6, 1518–25; Raghavan, B., et al. J. Med.
  • Tubulysin analog having the following formula (IV) :
  • R 1 , R 2 , R 3 , and R 4 are independently H, C 1 ⁇ C 8 alkyl; C 2 ⁇ C 8 heteroalkyl, or heterocyclic; C 3 ⁇ C 8 aryl, Ar-alkyl, cycloalkyl, alkylcycloalkyl, heterocycloalkyl, heteroalkylcycloalkyl, carbocyclic, or alkylcarbonyl; or R 1 R 2 , R 1 R 3 , R 2 R 3 , R 3 R 4 , R 5 R 6 , R 11 R 12 or R 13 R 14 form a 3 ⁇ 7 membered carbocyclic, cycloalkyl, heterocyclic, heterocycloalkyl, aromatic or heteroaromatic ring system; R 1 and R 2 can be independently absent when they link to L 1 or L 2 independently or simultaneously, Y 1 is N or CH;
  • R 5 , R 6 , R 8 , R 10 and R 11 are independently H, or C 1 ⁇ C 4 alkyl or heteroalkyl;
  • X 1 is O, S, S-S, NH, CH 2 or NR 14 ;
  • R 15 ⁇ R 16 and R 17 is independently H, C 1 ⁇ C 8 alkyl, heteroalkyl; C 2 -C 8 of alkenyl, alkynyl, heteroalkyl, heterocycloalkyl; C 3 -C 8 of aryl, Ar-alkyl, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, heteroaryl, alkylcarbonyl, or Na + , K + , Cs + , Li + , Ca 2+ , Mg + , Zn 2+ , N + (R 1 ) (R 2 ) (R 3 ) (R 4 ) , HN + (C 2 H 5 OH) 3 salt;
  • R 20 is H; C 1 -C 8 of linear or branched alkyl or heteroalkyl; C 2 -C 8 of linear or branched alkenyl, alkynyl, alkylcycloalkyl, heterocycloalkyl; C 3 -C 8 linear or branched of aryl, Ar-alkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, heteroaryl; carbonate (-C (O) OR 17 ) , carbamate (-C (O) NR 17 R 18 ) ; or 1-8 carbon atoms of carboxylate, esters, ether, or amide; or 1 ⁇ 8 amino acids; or polyethyleneoxy unit of formula (OCH 2 CH 2 ) p or (OCH 2 CH (CH 3 ) ) p , wherein p is an integer from 0 to about 1000; or R 20 is absent and the oxygene forms a ketone, or combination above thereof
  • Calicheamicins and their related enediyne antibiotics that are described in: Nicolaou, K. C. et al, Science 1992, 256, 1172-1178; Proc. Natl. Acad. Sci USA. 1993, 90, 5881-8) , U.S. Patent Nos. 4,970,198; 5,053,394; 5,108,912; 5,264,586; 5,384,412; 5,606,040; 5,712,374; 5,714,586; 5,739,116; 5,770,701; 5,770,710; 5,773,001; 5,877,296; 6,015,562; 6,124,310; 8,153,768.
  • Exemplary enediynes include, but are not limited to, calicheamicin, esperamicin, uncialamicin, dynemicin, and their derivatives.
  • the structure of calicheamicins is preferred the following formula:
  • Geldanamycins are benzoquinone ansamycin antibiotic that bind to Hsp90 (Heat Shock Protein 90) and have been used antitumor drugs.
  • exemplary geldanamycins include, but are not limited to, 17-AAG (17-N-Allylamino-17-Demethoxygeldanamycin) and 17-DMAG (17-Dimethylaminoethylamino-17-demethoxygeldanamycin) .
  • Maytansines or their derivatives maytansinoids inhibit cell proliferation by inhibiting the mcirotubules formation during mitosis through inhibition of polymerization of tubulin. See Remillard et al., Science 189: 1002-1005 (1975) .
  • Exemplary maytansines and maytansinoids include, but are not limited to, mertansines (DM1, DM4) , maytansinol and its derivatives as well as ansamitocin. Maytansinoids are described in U.S. Patent Nos.
  • camptothecin and its derivatives, which are topoisomerase inhibitors to prevent DNA re-ligation and therefore to causes DNA damage resulting in apoptosis, are described in: Shang, X.F. et al, Med Res Rev. 2018, 38 (3) : 775-828; Botella, P. and Rivero-Buceta, E. J Control Release. 2017, 247: 28-54; Martino, E. et al, Bioorg Med Chem Lett. 2017, 27 (4) : 701-707; Lu, A., et al, Acta Pharmacol Sin 2007, 28 (2) : 307–314.
  • Camptothecin CPT
  • R 1, R 2 and R 4 are independently selected from H, F, Cl, Br, CN, NO 2 , C 1 ⁇ C 8 alkyl; O-C 1 ⁇ C 8 alkyl; NH-C 1 ⁇ C 8 alkyl; C 2 -C 8 of heteroalkyl, alkylcycloalkyl, heterocycloalkyl; C 3 -C 8 of aryl, Ar-alkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, heteroaryl; or 2-8 carbon atoms of esters, ether, amide, carbonate, urea, or carbamate; R 3 is H, OH, NH 2 , C 1 ⁇
  • camptothecins are preferred the following formula:
  • P 1 is H, OH, NH 2 , COOH, C (O) NH 2 , OCH 2 OP (O) (OR 18 ) 2 , OC (O) OP (O) (OR 18 ) 2 , OPO (OR 18 ) 2 , NHPO (OR 18 ) 2 , OC (O) R 18 , OP (O) (OR 18 ) OP (O) (OR 18 ) 2 , OC (O) NHR 18 , OC (O) N (C 2 H 4 ) 2 NCH 3 , OSO 2 (OR 18 ) , O- (C 4 -C 12- glycoside) , OC (O) N (C 2 H 4 )
  • Combretastatins are natural phenols with vascular disruption properties in tumors.
  • Exemplary combretastatins and their derivatives include, but are not limited to, combretastatin A-4 (CA-4) , CA4- ⁇ Gals, CA-4PD, CA4-NPs and ombrabulin.
  • Taxanes which includes Paclitaxel (Taxol) , a cytotoxic natural product, and docetaxel (Taxotere) , a semi-synthetic derivative, and their analogs which are preferred for conjugation are exampled in: K C. Nicolaou et al., J. Am. Chem. Soc. 117, 2409-20, (1995) ; Ojima et al, J. Med. Chem. 39: 3889-3896 (1996) ; 40: 267-78 (1997) ; 45, 5620-3 (2002) ; Ojima et al., Proc. Natl. Acad. Sci., 96: 4256-61 (1999) ; Kim et al., Bull.
  • Ar and Ar’ are independently aryl or heteroaryl.
  • Anthracyclines are mammalian DNA topoisomerases II inhibitors that are able to stabilize enzyme-DNA complexes wherein DNA strands are cut and covalently linked to the antibody-like protein. These anticancer agents maintain a prominent role in treating many forms of solid tumors and acute leukemias during the last several decades.
  • anthracyclines cause cardiovascular morbidity and mortality (Sagi, J. C., et al, Pharmacogenomics. 2016, 17 (9) , 1075-87; McGowan, J. V., et al, Cardiovasc Drugs Ther. 2017, 31 (1) , 63-75) .
  • reasearchers actively are using the conjugation of anthracyclines to a cell-binding antibody, or antibody-like protein molecule as a general approach for improving the therapeutic index of these drugs, (Mollaev, M. et al, Int J Pharm. 2018 Dec 29. pii: S0378-5173 (18) 30991-8; Rossin, R., et al, Bioconjug Chem. 2016, 27 (7) : 1697-706; Dal Corso, A., et al, J Control Release. 2017, 264: 211-218) .
  • anthracyclines include, but are not limited to, daunorubicin, doxorubicin (i.e., adriamycin) , epirubicin, idarubicin, valrubicin, and mitoxantrone.
  • doxorubicin i.e., adriamycin
  • epirubicin i.e., adarubicin
  • valrubicin idarubicin
  • mitoxantrone i.e., mitoxantrone.
  • Vinca alkaloids are a set of anti-mitotic and anti-microtubule alkaloid agents that work by inhibiting the ability of cancer cells to divide.
  • Vinca alkaloids include vinblastine, vincristine, vindesine, leurosine, vinorelbine, catharanthine, vindoline, vincaminol,ieridine, minovincine, methoxyminovincine, minovincinine, vincadifformine, desoxyvincaminol, vincamajine, vincamine, vinpocetine, and vinburnine.
  • the structures of vinca alkaloids are preferred vinblastine, vincristine having the following formula:
  • Dolastatins and their peptidic analogs and derivatives, auristatins are highly potent antimitotic agents that have been shown to have anticancer and antifungal activity. See, e.g., U.S. Pat. No. 5,663,149 and Pettit et al., Antimicrob. Agents Chemother. 42: 2961-2965, 1998.
  • Exemplary dolastatins and auristatins include, but are not limited to, dolastatin 10, auristatin E (AE) , auristatin EB (AEB) , auristatin EFP (AEFP) , MMAD (Monomethyl Auristatin D or monomethyl dolastatin 10) , MMAF (Monomethyl Auristatin F or N-methylvaline-valine-dolaisoleuine-dolaproine-phenylalanine) , MMAE (Monomethyl Auristatin E or N-methylvaline-valine-dolaisoleuine-dolaproine-norephedrine) , 5-benzoylvaleric acid-AE ester (AEVB) , Auristatin F phenylene diamine (AFP) and other novel auristatins.
  • AE auristatin E
  • AEB auristatin EB
  • AEFP auristatin EFP
  • auristatin analogs are preferred the following formula (Ih-01) , (Ih-02) , (Ih-03) , (Ih-04) , (Ih-05) , (Ih-06) , (Ih-07) , (Ih-08) , (Ih-09) , (Ih-10) , and (Ih-11) :
  • R 1 , R 2 , R 3 , R 4 and R 5 are independently H; C 1 -C 8 linear or branched alkyl, aryl, heteroaryl, heteroalkyl, alkylcycloalkyl, ester, ether, amide, amines, heterocycloalkyl, or acyloxylamines; or peptides containing 1-8 aminoacids, or polyethyleneoxy unit having formula (OCH 2 CH 2 ) p or (OCH 2 CH (CH 3 ) ) p , wherein p is an integer from 1 to about 1000.
  • R 1 R 2 , R 2 R 3 , R 1 R 3 or R 3 R 4 together can form 3 ⁇ 8 member cyclic ring of alkyl, aryl, heteroaryl, heteroalkyl, or alkylcycloalkyl group;
  • Y 1 and Y 2 are independently O, NH, NHNH, NR 5 , S, C (O) O, C (O) NH, OC (O) NH, OC (O) O, NHC (O) NH, NHC (O) S, OC (O) N (R 1 ) , N (R 1 ) C (O) N (R 2 ) , C (O) NHNHC (O) and C (O) NR 1 when linked to the connecting site (that links to L 1 and/or L 2 independently) ; or OH, NH 2 , NHNH 2 , NHR 5 , SH, C (O) OH, C (O) NH 2 , OC (O) NH 2 , OC
  • Hemiasterlin and its analogues bind to the tubulin, disrupt normal microtubule dynamics, and, at stoichiometric amounts, depolymerize microtubules.
  • the structure of maytansinoids is preferred the following formula:
  • R 1 , R 2 , R 3 , R 4 and R 5 are independently H; C 1 -C 8 linear or branched alkyl, aryl, heteroaryl, heteroalkyl, alkylcycloalkyl, ester, ether, amide, amines, heterocycloalkyl, or acyloxylamines; or peptides containing 1-8 aminoacids, or polyethyleneoxy unit having formula (OCH 2 CH 2 ) p or (OCH 2 CH (CH 3 ) ) p , wherein p is an integer from 1 to about 5000;
  • R 2 R 3 can form 3 ⁇ 8 member cyclic ring of alkyl, aryl, heteroaryl, heteroalkyl, or alkylcycloalkyl group.
  • Eribulin which is binding predominantly to a small number of high affinity sites at the plus ends of existing microtubules has both cytotoxic and non-cytotoxic mechanisms of action. Its cytotoxic effects are related to its antimitotic activities, wherein apoptosis of cancer cells is induced following prolonged and irreversible mitotic blockade (Kuznetsov, G. et al, Cancer Research. 2004, 64 (16) : 5760–6.; Towle, M. J, et al, Cancer Research. 2010, 71 (2) : 496–505) .
  • Eribulin has been approved by US FDA for the treatment of metastatic breast cancer who have received at least two prior chemotherapy regimens for late-stage disease, including both anthracycline-and taxane-based chemotherapies, as well as for the treatment of liposarcoma (a specific type of soft tissue sarcoma) that cannot be removed by surgery (unresectable) or is advanced (metastatic) .
  • Eribulin has been used as payload for ADC conjugates (US20170252458) .
  • the structure of Eribulin is preferred the following formula, Eb01:
  • NAMPT nicotinamide phosphoribosyltransferases
  • NAD + acts as a coenzyme in redox reactions, as a donor of ADP-ribose moieties in ADP-ribosylation reactions, as a precursor of the second messenger molecule cyclic ADP-ribose, as well as acting as a substrate for bacterial DNA ligases and a group of enzymes called sirtuins that use NAD + to remove acetyl groups from proteins.
  • NAD + emerges as an adenine nucleotide that can be released from cells spontaneously and by regulated mechanisms (Smyth L. M, et al, J. Biol. Chem. 2004, 279 (47) , 48893–903; Billington R. A, et al, Mol Med.
  • NAMPT inhibitors are preferred the following formula, NP01, NP02, NP03, NP04, NP05, NP06, NP07, NP08, and NP09:
  • X 5 is F, Cl, Br, I, OH, OR 1 , R 1 , OPO 3 H 2 , OSO 3 H, NHR 1 , OCOR 1 , NHCOR 1 .
  • a benzodiazepine dimer and its analogs are anti-tumor agents that contain one or more immine functional groups, or their equivalents, that bind to duplex DNA.
  • PBD and IGN molecules are based on the natural product athramycin, and interact with DNA in a sequence-selective manner, with a preference for purine-guanine-purine sequences.
  • the preferred benzodiazepine dimers according to the present invention are exampled in: US Patent Nos. 8, 163, 736; 8, 153, 627; 8, 034, 808; 7, 834, 005; 7, 741, 319; 7, 704, 924; 7, 691, 848; 7, 678, 787; 7, 612, 062; 7, 608, 615; 7, 557, 099; 7, 528, 128; 7, 528, 126; 7, 511, 032; 7, 429, 658; 7, 407, 951; 7, 326, 700; 7, 312, 210; 7, 265, 105; 7, 202, 239; 7, 189, 710; 7, 173, 026; 7, 109, 193; 7, 067, 511; 7, 064, 120; 7, 056, 913; 7, 049, 311; 7, 022, 699; 7, 015, 215; 6, 979, 684; 6, 951, 853; 6, 884, 799; 6, 800, 622; 6, 747, 144; 6, 660
  • X 1 , X 2 , Y 1 , Y 2 , Z 1 , Z 2 , and n are defined the same above;
  • X 1 , X 2 , Y 1 and Y 2 are independently O, N, NH, NHNH, NR 5 , S, C (O) O, C (O) NH, OC (O) NH, OC (O) O, NHC (O) NH, NHC (O) S, OC (O) N (R 1 ) , N (R 1 ) C (O) N (R 1 ) , CH, C (O) NHNHC (O) and C (O) NR 1 ; R 1 , R 2 , R 3 , R
  • NR 5 R 5 ’ heterocycloalkyl, or acyloxylamines (-C (O) NHOH, -ONHC (O) R 5 ) ; or peptides containing 1-20 natural or unnatural aminoacids, or polyethyleneoxy unit of formula (OCH 2 CH 2 ) p or (OCH 2 CH (CH 3 ) ) p , wherein p is an integer from 1 to about 5000.
  • R 1 R 2 , R 2 R 3 , R 1 R 3 , R 1’ R 2’ , R 2’ R 3’ , or R 1’ R 3’ can independently form 3 ⁇ 8 member cyclic ring of alkyl, aryl, heteroaryl, heteroalkyl, or alkylcycloalkyl group;
  • X 3 and Y 3 are independently N, NH, CH 2 or CR 5 , or one of X 3 and Y 3 can be absent;
  • M 1 and M 2 are independently H, Na, K, Ca, Mg, NH 4 , NR 1 R 2 R 3 ;
  • X 6 is CH, N, P (O) NH, P (O) NR 1 , CHC (O) NH, C 3 -C 8 aryl, heteroaryl, alkylcycloalkyl, acyloxyl, alkylaryl, alkylaryloxyl, alkylarylamino, or an Aa (amino acid, is preferably selected from Lys, Phe, Asp, Glu, Ser, Thr, His, Cys, Tyr, Trp, Gln, Asn, Arg) ; is defined the same above.
  • CC-1065 analog and doucarmycin analogs are also preferred to be used for a conjugate of the present process invention.
  • the examples of the CC-1065 analogues and doucarmycin analogs as well as their synthesis are described in: e.g. Warpehoski, et al, J. Med. Chem. 31: 590-603 (1988) ; D. Boger et al., J. Org. Chem; 66; 6654-61, 2001; U.S.
  • X 1 , X 2 , Y 1 and Y 2 are independently O, NH, NHNH, NR 5 , S, C (O) O, C (O) NH, OC (O) NH, OC (O) O, NHC (O) NH, NHC (O) S, OC (O) N (R 1 ) , N (R 1 ) C (O) N (R 2 ) , C (O) NHNHC (O) and C (O) NR 1 when linked to the connecting site or OH, NH 2 , NHNH 2 , NHR 1 , SH, C (O) OH, C (O) NH 2 , OC (O) NH 2 , OC (O) OH, NHC (O) NH 2 , NHC (O) SH, OC (O) NH (R 1 ) , N (R 1 ) C (O) NH (O) NH (R 2 ) , C (O) NHNHC (
  • amatoxin and its analogs which are a subgroup of at least ten toxic compounds originally found in several genera of poisonous mushrooms, most notably Amanita phalloides and several other mushroom species, are also preferred for conjugation of the present patent.
  • These ten amatoxins named ⁇ -Amanitin, ⁇ -Amanitin, ⁇ -Amanitin, ⁇ -Amanitin, Amanullin, Amanullinic acid, Amaninamide, Amanin, Proamanullin, are rigid bicyclic peptides that are synthesized as 35-amino-acid proproteins, from which the final eight amino acids are cleaved by a prolyl oligopeptidase (Litten, W.
  • Spliceostatins and pladienolides are anti-tumor compounds which inhibit splicing and interacts with spliceosome, SF3b.
  • spliceostatins include, but are not limited to, spliceostatin A, FR901464, and (2S, 3Z) -5- ⁇ [ (2R, 3R, 5S, 6S) -6- ⁇ (2E, 4E) -5- [ (3R, 4R, 5R, 7S) -7- (2-hydrazinyl-2-oxoethyl) -4-hydroxy-1, 6-dioxaspiro [2.5] oct-5-yl] -3-methylpenta-2, 4-dien-1-y-l ⁇ -2, 5-dimethyltetrahydro-2H-pyran-3-yl] amino ⁇ -5-oxopent-3-en-2-yl acetate having the core structure:
  • pladienolides examples include, but are not limited to, Pladienolide B, Pladienolide D, and E7107.
  • Protein kinase inhibitors that block the action of an enzyme to add a phosphate (PO 4 ) group to serine, threonine, or tyrosine amino acids on an antibody-like protein, and can modulate the protein function.
  • the protein kinase inhibitors can be used to treat diseases due to hyperactive protein kinases (including mutant or overexpressed kinases) in cancer or to modulate cell functions to overcome other disease drivers.
  • protein kinase inhibitors are preferred to selected from Adavosertib, Afatinib, Axitinib, Bafetinib, Bosutinib, Cobimetinib, Crizotinib, Cabozantinib, Dasatinib, Entrectinib, Erdafitinib, Erlotinib, Erlotinib, Fostamatinib, Gefitinib, Ibrutinib, Imatinib, Lapatinib, Lenvatinib, Mubritinib, Nilotinib, Pazopanib, Pegaptanib, Ponatinib, Rebastinib, Regorafenib, Ruxolitinib, Sorafenib, Sunitinib, SU6656, Tofacitinib, Vandetanib, Vemurafenib, Entrectinib, Palbociclib, Ribociclib, Riboc
  • Z 5 and Z 5 ’ are independently selected from O, NH, NHNH, NR 5 , S, C (O) O, C (O) NH, OC (O) NH, OC (O) O, NHC (O) O, NHC (O) NH, NHC (O) S, OC (O) N (R 1 ) , N (R 1 ) C (O) N (R 2 ) , C (O) NHNHC (O) and C (O) NR 1 .
  • a MEK inhibitor inhibits the mitogen-activated protein kinases MEK1 and/or MEK2 which is often overactive in some cancers.
  • MEK inhibitors are especially used for treatment of BRAF-mutated melanoma, and KRAS/BRAF mutated colorectal cancer, breast cancer, and non-small cell lung cancer (NSCLC) .
  • MEK inhibitors are selected from PD0325901, selumetinib (AZD6244) , cobimetinib (XL518) , refametinib, trametinib (GSK1120212) , pimasertib, Binimetinib (MEK162) , AZD8330, RO4987655, RO5126766, WX-554, E6201, GDC-0623, PD-325901 and TAK-733.
  • the preferred MEK inhibitors are selected from Trametinib (GSK1120212) , Cobimetinib (XL518) , Binimetinib (MEK162) , selumetinib having the following formula:
  • Z 5 is selected from O, NH, NHNH, NR 5 , S, C (O) O, C (O) NH, OC (O) NH, OC (O) O, NHC (O) O, NHC (O) NH, NHC (O) S, OC (O) N (R 1 ) , N (R 1 ) C (O) N (R 2 ) , C (O) NHNHC (O) and C (O) NR 1 ;
  • a proteinase inhibitor that are used as a payload is preferably selected from: Carfilzomib, Clindamycin, Rumblemulin, Indibulin, as shown in the following structures:
  • An immunotoxin herein is a macromolecular drug which is usually a cytotoxic protein derived from a bacterial or plant protein, such as Diphtheria toxin (DT) , Cholera toxin (CT) , Trichosanthin (TCS) , Dianthin, Pseudomonas exotoxin A (ETA′) , Erythrogenic toxins, Diphtheria toxin, AB toxins, Type III exotoxins, etc. It also can be a highly toxic bacterial pore-forming protoxin that requires proteolytic processing for activation. An example of this protoxin is proaerolysin and its genetically modified form, topsalysin.
  • Topsalysin is a modified recombinant protein that has been engineered to be selectively activated by an enzyme in the prostate, leading to localized cell death and tissue disruption without damaging neighboring tissue and nerves;
  • An immunotoxin herein is preferably conjugated via the process of the application through an amino acid having free amino, thiol or carboxyl acid group; and more preferably through N-terminal amino acid.
  • a certain cell receptor agonist, a cell stimulating molecule or intracellular signalling molecule can be as a drug D conjugated via the process of the invention.
  • a cell-binding ligand or receptor agonist selected from: Folate derivatives; Glutamic acid urea derivatives; Somatostatin and its analogs (selected from the group consisting of octreotide (Sandostatin) and lanreotide (Somatuline) ) ; Aromatic sulfonamides; Pituitary adenylate cyclase activating peptides (PACAP) (PAC1) ; Vasoactive intestinal peptides (VIP/PACAP) (VPAC1, VPAC2) ; Melanocyte-stimulating hormones ( ⁇ -MSH) ; Cholecystokinins (CCK) /gastrin receptor agonists; Bombesins (selected from the group consisting of Pyr-Gln-Arg-Leu-Gly-Asn-Gln-Trp-Ala-Val-Gly-His-Leu-Met-NH 2 ) /gastrin-releasing peptide (GR
  • a cell-binding molecule/ligand or a cell receptor agonist selected from the following: LB01 (Folate) , LB02 (PMSA ligand) , LB03 (PMSA ligand) , LB04 (PMSA ligand) , LB05 (Somatostatin) , LB06 (Somatostatin) , LB07 (Octreotide, a Somatostatin analog) , LB08 (Lanreotide, a Somatostatin analog) , LB09 (Vapreotide (Sanvar) , a Somatostatin analog) , LB10 (CAIX ligand) , LB11 (CAIX ligand) , LB12 (Gastrin releasing peptide receptor (GRPr) , MBA) , LB13 (luteinizing hormone-releasing hormone (LH-RH) ligand and GnRH) , LB14 (luteinizing hormone-releasing hormone (LH-R
  • Y 5 is N, CH, C (Cl) , C (CH 3 ) , or C (COOR 1 ) ;
  • R 12 is H, C 1 -C 6 Alkyl, C 3 -C 8 Ar;
  • X 4 , and Y 1 are independently O, NH, NHNH, NR 1 , S, C (O) O, C (O) NH, OC (O) NH, OC(O) O, NHC (O) NH, NHC (O) S, OC (O) N (R 1 ) , N (R 1 ) C (O) N (R 1 ) , CH 2, C (O) NHNHC (O) and C (O) NR 1 .
  • one, two or more DNA, RNA, mRNA, small interfering RNA (siRNA) , microRNA (miRNA) , and PIWI interacting RNAs (piRNA) can be as a drug conjugated via the process of the invention:
  • X 1 , and Y are independently O, NH, NHNH, NR 1 , S, C (O) O, C (O) NH, OC (O) NH, OC (O) O, NHC (O) NH, NHC (O) S, OC (O) N (R 1 ) , N (R 1 ) C (O) N (R 1 ) , CH 2, C (O) NHNHC (O) and C (O) NR 1 .
  • the oxidant which can be added in step (c) (after the step of the conjugation reaction) in the process of invention is preferably selected from dehydroascorbic acid (DHAA) to re-oxidize unreacted thiol groups, thus leading to restore the disulfide linkage in the antibody or antibody-like protein for having longer half life.
  • DHAA dehydroascorbic acid
  • the concentration of the oxidant in the reaction solution may be 0.01 mM -1.0 mM.
  • an excess amount of disulfide compound such as cystine can be added in the step (c) to replace DHAA.
  • the disulfide compound can be reduced by the excess reductant, such as TCEP in step (b) , to form a thiol compound, which simultaneously reacts to the excessive conjugation linker or linker/payload complex containing thiol reactive groups (e.g. maleimide) , and following by removing of the generated thiol-succinimide linker/payload complex by chromatography.
  • thiol reactive groups e.g. maleimide
  • n is 1 –20; n’ is 1-10; preferably n is 1 -8 and n’ is 1 -4; more preferably n is 2 -4 and n’ is 1 -2; D 1 , D 2 , L 1 , L 2 , and E 1 are described the same above; S (sulfur) is generated from the reduction of disulfide bonds in the antibody-like protein (e.g. antibody) under process of the invention; mAb is an antibody-like protein;
  • Lv 1 ’ and Lv 2 ’ are independently the resulting groups that a thiol in mAb reacted with Lv 1 and Lv 2 , whose structures described above.
  • Lv 1 ’ and Lv 2 ’ are independently having the following structures:
  • mAb is an antibody-like protein, preferably an antibody.
  • the conjugates are spefically linked to the tiols between heavy-light chains of the antibody when an antibody-like protein is specifically an antibody.
  • a linker having formula (VIII) , (IX) or (X) illustrated below can react first to the selectively reduced thiols in the antibody or antibody-like protein (e.g. typically thiols between heavy-light chain when the antibody or antibody-like protein is IgG antibody) independently, followed by condensation with a cytotoxic drug or cytotoxic drug/linker complex to form the conjugates of formula (V) , (VI) , or (VII) as shown above:
  • L 1 , L 2 , E 1 , Lv 1 , and Lv 2 are defined the same above for Formula (I) , (II) and (III) ; wherein Lv 5 and Lv 6 are independently selected from
  • X 1 ’ is F, Cl, Br, I, OTs (tosylate) , OTf (triflate) , OMs (mesylate) , OC 6 H 4 (NO 2 ) , OC 6 H 3 (NO 2 ) 2 , OC 6 F 5 , OC 6 HF 4 , or Lv 3 ;
  • X 2 ’ is O, NH, N (R 1 ) , or CH 2 ;
  • R 3 and R 5 are independently H, R 1 , aromatic, heteroaromatic, or aromatic group wherein one or several H atoms are replaced independently by -R 1 , -halogen, -OR 1 , -SR 1 , -NR 1 R 2 , -NO 2 , -S (O) R 1 , -S (O) 2 R 1, or -COOR 1 ;
  • Lv 3 and Lv 3 ’ are independently a leaving group selected from F, Cl, Br, I, nitrophenoxyl; N-
  • acetyl anhydride formyl anhydride; or an intermediate molecule generated with a condensation reagent for peptide coupling reactions or for Mitsunobu reactions; wherein the fuction groups Lv 5 and/or Lv 6 can be also reacted with a thiol in a cytotoxic drug as long as the reaction are at least one fold faster or slower than the reaction between Lv 1 or Lv 2 and a thiol in an antibody-like protein, in particular, in an antibody.
  • linker of formula (VIII) , (IX) or (X) illustrated above can react first with a cytotoxic drug to form the cytotoxic drug/linker complex molecule of formula (I) , (II) or (III) , follow by reaction with the reduced thiols in the antibody or antibody-like protein independently to form the conjugate of formula (V) , (VI) , or (VII) under process of this invention.
  • the first step condensation reaction of the formula (VIII) , (IX) or (X) to a cytotoxic drug can be in a separated pot, and the resulted cytotoxic drug/linker complex molecules of formula (I) , (II) or (III) can be optionally purified by a chromatography, extraction or precipitatation before for conjugation to the reduced thiols in the antibody-like protein.
  • the first step of specific reduction of disulfide bonds in an antibody-like protein and conjugation reaction with formula (I) , (II) or (III) are preferred in the same pot without separation of intermidiates.
  • each step of the reactions for the linker of formula (VIII) , (IX) or (X) can be conducted at different conditions in the same or different reaction pots.
  • a drug containing an amino group can undergo condensation with a carboxylic acid group in the linker in the present of a condensation regent, e.g. EDC, TBTU or BroP, to give a modified drug/linker complex of Formula (I) , (II) or III) bearing amide bonds.
  • This condensation reaction can be performed at physiological buffer solution wherein the carboxylic acid group at one terminal of the linker of formula (VIII) , (IX) or (X) is activated to be N-hydoxylsuccinimidyl (NHS) , pentfluorophenyl, dinitrophenyl ester, or carboxylic acid chloride group, etc, which can react to a drug bearing an amino group to provide drug/linker complex of Formula (I) , (II) or III) , then subsequently or simultaneously undergo the conjugation to thiols of an antibody-like protein according to the process of the present application to form the conjugate of formula (V) , (VI) , or (VII) .
  • a thiol reactive group e.g. maleimido, vinylsulfonyl, haloacetyl, acrylic, substituted propiolic
  • a drug reactive group e.g. hydoxylsuccinimidyl (NHS) , pentfluorophenyl, dinitrophenyl ester, amino, alkyloxylamino or clickable chemistry group (e.g.
  • a drug bearing a reactive group matched to the reactive group in the antibody-like protein-linker conjugate of formula (XI) , (XII) or (XIII) accordingly can be subsequently or simultaneously added to the reaction solution to provide the conjugate of formula (V) , (VI) , or (VII) .
  • the antibody-like protein-linker conjugate of formula (XI) , (XII) or (XIII) can be optionally purified before proceeding the condensation with a drug, and the condensation condition of the second step can be adjusted, e.g. the pH is adjusted to 6.5 –8.0, and/or temperature is adjusted to 20 -45 °C.
  • the antibody-like protein can be modified through attachment of a heterobifunctional cross linker of formula (XI) , (XII) or (XIII) , such as with linkers of Amine-to-Sulfhydryl (succinimidyl (NHS) ester/maleimide, NHS ester/pyridyldithiol, NHS esters/haloacetyl) , diazirine (SDA) –to-Sulfhydryl, Azide-to-Sulfhydryl, Alkyne-to-Sulfhydryl, Sulfhydryl-to-Carbohydrate (Maleimide/Hydrazide, Pyridyldithiol/Hydrazide, haloacetyl/Hydrazide) , Hydroxyl-to-Sulfhydryl (Isocyanate/Maleimide/N-(trimethyl)
  • the reactive group of a drug/cytotoxic agent that reacting to a modified an antibody-linker conjugate of formula (XI) , (XII) or (XIII) to give the final conjugate can be in different ways accordingly.
  • the conjugate linked via disulfide bonds is achieved via the first step, a linker of formula (VIII) , (IX) or (X) is conjugated to the antibody-like protein at 2 °C -8 °C, pH 4.5 –6.0, according to the present invention of reduction and conjugation of an antibody-like protein, following by a disulfide exchange between a drug containing a free thiol group and the disulfide bond ( (e.g.
  • the excess reduction agent e.g. TCEP, or tri (3-hydroxylpropyl) phosphine
  • TCEP tri (3-hydroxylpropyl) phosphine
  • Synthesis of the conjugates linked via thioether is achieved by first reaction of a linker containing both thiol reactive terminals of maleimido or haloacetyl or ethylsulfonyl or substituted propiolic group to the thiols in an antibody which are reduced by the process of the present patent application at 2 °C -8 °C, pH 4.5 –6.0 to give the antibody-linker conjugate of formula (XI) , (XII) or (XIII) , following by reaction of a drug containing a thiol at pH 6.5 –8.0, at 20 °C -40 °C to to provide the conjugate of formula (V) , (VI) , or (VII) .
  • the preferred methods of synthesis of the disulfide or thiol-ether linked conjugates are through the first chemical synthesis the drug-linker complex having disulfide or thiol-ether bonds of the formula (I) , (II) or (III) ; following by reaction with the thiols in the protein (antibody) according the process of the invention.
  • Synthesis of conjugates bearing an acid labile hydrazone linkage can be achieved by reaction of a carbonyl group with the hydrazide moiety in the linker, by methods known in the art (see, for example, P. Hamann et al., Cancer Res. 53, 3336-34, 1993; B. Laguzza et al., J. Med. Chem., 32; 548-55, 1959; P. Trail et al., Cancer Res., 57; 100-5, 1997) .
  • Synthesis of conjugates bearing triazole linkage can be achieved by reaction of a 1-yne group of the drug with the azido moiety in the linker, through the click chemistry (Huisgen cycloaddition) (Lutz, J-F.
  • Synthesis of the conjugates linked via oxime is achieved by reaction of a modified antibody-like protein containing a ketone or aldehyde and a drug containing oxyamine group.
  • a drug bearing a hydroxyl group or a thiol group can be reacted with a modified linker of Formula (XI) , (XII) , or (XIII) , bearing a halogen, particularly the alpha halide of carboxylates, in the presence of a mild base, e.g.
  • a drug containing a hydroxyl group can be condensed with a linker of Formula (XI) , (XII) , or (XIII) bearing a carboxyl group, in the presence of a dehydrating agent, such as EDC or DCC, to give ester linkage, then the subject drug/linker complex undergoes the conjugation with an antibody-like protein under the process of the present invention.
  • a dehydrating agent such as EDC or DCC
  • a drug containing an amino group can condensate with a carboxyl ester of NHS, imidazole, nitrophenoxyl; N-hydroxysuccinimide (NHS) ; methylsufonylphenoxyl; dinitrophenoxyl; pentafluorophenoxyl; tetrafluorophenoxyl; difluorophenoxyl; monofluorophenoxyl; pentachlorophenoxyl; triflate; imidazole; dichlorophenoxyl; tetrachlorophenoxyl; 1-hydroxyben- zotriazole; tosylate; mesylate; 2-ethyl-5-phenylisoxazolium-3′-sulfonate on the antibody-like protein-linker of Formula (VIII) , or (XI) to give a conjugate via amide bond linkage of Formula (V) , (VI) , or (VII) .
  • NHS N-hydroxysuccinimide
  • the resulted conjugates of formula (V) , (VI) , or (VII) are over 75%linked to the cysteine sites between heavy-light chains of an antibody, and are less than 15%linked to the cysteine sites between heavy-heavy chains (hinge region) of an antibody.
  • the resulted conjugate may be purified by standard biochemical means, such as gel filtration on a Sephadex G25 or Sephacryl S300 column, adsorption chromatography, ion (cation or anion) exchange chromatography or by dialysis (ultrafiltration or hyperfiltration (UF) and diafiltration (DF) ) .
  • a small size molecule of antibody-like protein (e.g. ⁇ 10 KD) conjugated with a small molecular drugs can be purified by chromatography such as by HPLC, medium pressure column chromatography or ion exchange chromatography.
  • the conjugate of Formula (V) , (VI) , or (VII) is preferably generated from a drug/linker complex of Formula (I) , (XII) , or (XIII) , as in a one pot reaction.
  • the Ellman reagent can be optionally used to monitor the efficient reduction of the disulfide bonds and conjugation of the tiols through measurement of the numbers of the free thiols during the reactions.
  • a UV spectrometry at wavelength of range 190-390 nm, preferably at 240-380 nm, more preferably at 240-330 nm is preferred to be used in assisting the reaction (via monitoring the conjugation) .
  • the conjugation reaction can be thus measured or conducted in a quartz cell or Pyrex flask in temperature control environment.
  • the drug/protein (antibody) ratios (DAR) of the conjugates can also be measured by UV at wavelength of range 240-380 nm via calculation of the concentrations of the drug and the protein, by Hydrophobic Interaction Chromatography (HIC-HPLC) via measurement of the integration areas of each drug/protein fragment, by Capilary electrophoresis (CE) , and/or by LC-MS or LC-MS/MS or CE-MS (the combination of liquid chromatography (LC) or CE with mass spectrometry (MS) via measurement of both the integration areas of LC or CE and Peak intensity of MS for each drug/protein fragment) .
  • HIC-HPLC Hydrophobic Interaction Chromatography
  • CE Capilary electrophoresis
  • CE-MS the combination of liquid chromatography (LC) or CE with mass spectrometry (MS) via measurement of both the integration areas of LC or CE and Peak intensity of MS for each drug/protein fragment
  • a drug or a drug/linker complex when a drug or a drug/linker complex is not well soluble in a water based buffer solution, up to 30%of water mixable (miscible) organic solvents, such as DMA, DMF, ethanol, methanol, acetone, acetonitrile, THF, isopropanol, dioxane, propylene glycol, or ethylene diol can be added as the co-solvent in water based buffer solution.
  • water mixable organic solvents such as DMA, DMF, ethanol, methanol, acetone, acetonitrile, THF, isopropanol, dioxane, propylene glycol, or ethylene diol
  • aqueous solutions for the modification of the antibody-like protein are buffered between pH 4 and 9, preferably between 6.0 and 7.5 and can contain any non-nucleophilic buffer salts useful for these pH ranges.
  • Typical buffers include phosphate, acetate, triethanolamine HCl, HEPES, and MOPS buffers, which can contain additional components, such as cyclodextrins, sucrose and salts, for examples, NaCl and KCl.
  • Other biological buffers that are used for the conjugation process are listed in the definition section.
  • the progress of the reaction can be monitored by measuring the decrease in the absorption at a certain UV wavelength, such as at 254 nm, or increase in the absorption at a certain UV wavelength, such as 280 nm, or the other appropriate wavelength.
  • isolation of the modified cell-binding antibody-like protein agent can be performed in a routine way, using for example gel filtration chromatography, or adsorptive chromatography.
  • the extent of the modification can be assessed by measuring the absorbance of the nitropyridine thione, dinitropyridine dithione, pyridine thione, carboxylamidopyridine dithione and dicarboxyl-amidopyridine dithione group released via UV spectra.
  • the modification or conjugation reaction can be monitored by LC-MS, preferably by UPLC-QTOF mass spectrometry, or Capilary electrophoresis–mass spectrometry (CE-MS) .
  • the linker compounds have diverse functional groups that can react with drugs, preferably cytotoxic agents that possess a suitable substituent.
  • the modified antibody-like protein bearing an amino or hydroxyl substituent can react with drugs bearing an N-hydroxysuccinimide (NHS) ester
  • the modified antibody-like protein bearing a thiol substituent can react with drugs bearing a maleimido or haloacetyl group
  • the modified antibody-like protein bearing a carbonyl (ketone or aldehyde) substituent can react with drugs bearing a hydrazide or an alkoxyamine.
  • One skilled in the art can readily determine which linker to use based on the known reactivity of the available functional group on the linkers.
  • p. p 1 , p 2 , and p 3 are independently 0 -100; m, m 1 , and m 2 are independently 0-20; n is 1 -10;
  • P 1 is H, OH, NH 2 , COOH, C (O) NH 2 , OCH 2 OP (O) (OR 18 ) 2 , OC (O) OP (O) (OR 18 ) 2 , OPO (OR 18 ) 2 , NHPO (OR 18 ) 2 , OC (O) R 18 , OP (O) (OR 18 ) OP (O) (OR 18 ) 2 , OC (O) NHR 18 , OC (O) N (C 2 H 4 ) 2 NCH 3 , OSO 2 (OR 18 ) , O- (C 4 -C 12- glycoside) , OC (O) N (C 2 H 4 ) 2 CH 2 N (C 2 H 4 ) 2 CH 3 , O- (C 1 -C 8 of linear or branched alkyl) , C 1 -C 8 of linear or branched alkyl or heteroalkyl; C 2 -C 8 of linear or branched alkenyl, al
  • R 1 , R 2 , R 3 , R 1’ , R 2’ , R 3’ , and R 4 are independently H, C 1 ⁇ C 8 alkyl; C 2 ⁇ C 8 heteroalkyl, or heterocyclic; C 3 ⁇ C 8 aryl, Ar-alkyl, cycloalkyl, alkylcycloalkyl, heterocycloalkyl, heteroalkylcycloalkyl, carbocyclic, or alkylcarbonyl; or R 1 R 2 , R 1 R 3 , R 2 R 3 , R 3 R 4 , R 1’ R 2’ , R 1’ R 3’ or R 2’ R 3’ form a 3 ⁇ 7 membered carbocyclic, cycloalkyl, heterocyclic, heterocycloalkyl, aromatic or heteroaromatic ring system;
  • R 7 , R 8 , and R 9 are independently H, OH, OR 1 , NH 2 , NHR 1 , C 1 -C 6 alkyl, or absent;
  • R 10 is CH 2 , O, NH, NR 1 , NHC (O) , NHC (O) NH, NHC (O) O, OC (O) O, C (O) , OC (O) , OC (O) (NR 1 ) , (NR 1 ) C (O) (NR 1 ) , C (O) R 1 or absent;
  • R 11 is OH, NH 2 , NHR 1 , NHNH 2 , NHNHCOOH, O-R 1 -COOH, NH-R 1 -COOH, NH- (Aa) r COOH, O (CH 2 CH 2 O) p CH 2 CH 2 OH, O (CH 2 CH 2 O) p CH 2 CH 2 NH 2 , NH (CH 2 CH 2 O) p CH 2 CH 2 NH 2 , NR 1 R 2 , O (CH 2 CH 2 O) p CH 2 CH 2 -COOH, NH (CH 2 CH 2 O) p CH 2 CH 2 COOH, NH-Ar-COOH, NH-Ar-NH 2 , O (CH 2 CH 2 O) p CH 2 CH 2 -NHSO 3 H, NH (CH 2 CH 2 O) p CH 2 CH 2 -NHSO 3 H, NH (CH 2 CH 2 O) p CH 2 CH 2 NHSO 3 H, R 1 -NHSO 3 H, NH-R
  • R 25 , R 26 and R 25 ’ are are independently H, Ac, R 1 , C (O) NHR 1 , C (O) R 1 , R 1 COOH, R 1 COOR 2 , R 1 OR 2 , R 1 CONHR 2 , CH 2 OAc, CH 2 NHAc, R 1 NH 2 , NR 1 R 2, N + R 1 R 2 R 3 , CH 2 CONH (CH 2 ) q1 COOH, CH2CONH (CH 2 ) q1 COOR 1 , CH 2 CONH (CH 2 ) q1 N + R 1 R 2 R 3 , , or (Aa) r;
  • X 1 , X 2 , X 4 , Y 1 and Y 2 are independently O, NH, NHNH, NR 5 , S, C (O) O, C (O) NH, OC (O) NH, OC (O) O, NHC (O) NH, NHC (O) S, OC (O) N (R 1 ) , N (R 1 ) C (O) N (R 2 ) , CH 2 , CHNH, CH 2 O, C (O) NHNHC (O) , OCH 2 C 6 H 4 NH, NHCH 2 C 6 H 4 NH, SCH 2 C 6 H 4 NH and C (O) NR 1 when linked to the connecting site or OH, NH 2 , NHNH 2 , NHR 1 , SH, C (O) OH, C (O) NH 2 , OC (O) NH 2 , OC (O) OH, NHC (O) NH 2 , NHC (O) SH
  • X 3 is H, CH 3 or X 1 ’R 1 ’, wherein X 1 ’ is NH, N (CH 3 ) , NHNH, O, or S; and R 1 ’ is H, C 1 -C 8 linear or branched alkyl, C 3 -C 8 aryl, heteroaryl, heteroalkyl, alkylcycloalkyl, acyloxylamines;
  • Z 3 ’ is H, COOR 1 , NH 2 , NHR 1 , OR 1 , CONHR 1 , NHCOR 1 , OCOR 1 , OP (O) (OM 1 ) (OM 2 ) , OCH 2 OP (O) (OM 1 ) (OM 2 ) , OSO 3 M 1 , R 1 , or O-glycoside (glucoside, galactoside, mannoside, glucuronoside/glucuronide, alloside, fructoside, etc. ) , NH-glycoside, S-glycoside or CH 2 -glycoside; M 1 and M 2 are independently H, Na, K, Ca, Mg, NH 4 , NR 1 R 2 R 3 ;
  • X 5 is F, Cl, Br, I, OH, OR 1 , R 1 , OPO 3 H 2 , OSO 3 H, NHR 1 , OCOR 1 , NHCOR 1 , CN or OCH 2 OP (O) (OM 1 ) (OM 2 ) ;
  • X 6 and Y 6 are independently CH, C (O) , N, P (O) NH, P (O) NR 1 , CHC (O) NH, C 1 -C 8 linear or branched alkyl, or heteroalkyl; C 3 -C 8 aryl, heteroaryl, alkylcycloalkyl, acyloxyl, alkylaryl, alkylaryloxyl, alkylarylamino, or an Aa (amino acid, preferably selected from Lys, Phe, Asp, Glu, Ser, Thr, His, Cys, Tyr, Trp, Gln, Asn, Arg) ;
  • Z 5 and Z 5 ’ are independently selected from O, NH, NHNH, NR 5 , S, C (O) O, C (O) NH, OC (O) NH, OC (O) O, NHC (O) O, NHC (O) NH, NHC (O) S, OC (O) N (R 1 ) , N (R 1 ) C (O) N (R 2 ) , C (O) NHNHC (O) or C (O) NR 1 ;
  • X 8 is O, S, NH, NHNH, NHR 1 , SR 12 , SSR 12 , SSCH (CH 3 ) R 1 , SSC (CH 3 ) 2 R 1 , or R 1;
  • R 1 , R 2 and R 3 are in dependently H, C 1 -C 8 linear or branched alkyl, C 3 -C 8 aryl, heteroaryl, heteroalkyl, alkylcycloalkyl, acyloxylamines;
  • Lv 1 is a leaving group defined the same above.
  • Lv 1 is selected from F, Cl, Br, I, OTs, OMS, OC 6 H 3 (NO 2 ) 2 , OC 6 F 5 , OC 6 H 4 (NO 2 ) , OC 6 Cl 5 ;
  • M 1 and M 2 are independently H, Na, K, Ca, Mg, NH 4 , NR 1 R 2 R 3 ; is defined the same above.
  • the antibody-like protein used for the conjugation process is proferred a cell-binding antibody-like protein molecule that binds to, complexes with, or reacts with a moiety of a cell population sought to be therapeutically or otherwise biologically modified.
  • antibody-like protein should be understood to include “antibody-like protein and peptide” except where the context requires otherwise.
  • Suitable antibody-like proteins which may be present in the conjugates of the invention include for example peptides, polypeptides, antibodies, antibody fragments, enzymes, cytokines, chemokines, receptors, blood factors, peptide hormones, toxin, transcription antibody-like proteins, or multimeric antibody-like proteins, wherein they have interchain disulfide bonds structurally.
  • Enzymes include carbohydrate-specific enzymes, proteolytic enzymes and the like, for example the oxidoreductases, transferases, hydrolases, lyases, isomerases and ligases disclosed by U.S. Pat. No. 4,179,337.
  • Specific enzymes of interest include asparaginase, arginase, adenosine deaminase, superoxide dismutase, catalase, chymotrypsin, lipase, uricase, bilirubin oxidase, glucose oxidase, glucuronidase, galactosidase, glucocerbrosidase, and glutaminase.
  • Blood antibody-like proteins include albumin, transferrin, Factor VII, Factor VIII or Factor IX, von Willebrand factor, insulin, ACTH, glucagen, somatostatin, somatotropins, thymosin, parathyroid hormone, pigmentary hormones, somatomedins, erythropoietin, luteinizing hormone, hypothalamic releasing factors, antidiuretic hormones, prolactin, interleukins, interferons, for example IFN- ⁇ . or IFN- ⁇ , colony stimulating factors, haemoglobin, cytokines, antibodies, antibody fragments, chorionicgonadotropin, follicle-stimulating hormone, thyroid stimulating hormone and tissue plasminogen activator.
  • allergen antibody-like proteins of interest are allergen antibody-like proteins disclosed by Dreborg et al Crit. Rev. Therap. Drug Carrier Syst. (1990) 6 315-365 as having reduced allergenicity when conjugated with a polymer such as poly (alkylene oxide) and consequently are suitable for use as tolerance inducers.
  • allergens disclosed are Ragweed antigen E, honeybee venom, mite allergen and the like.
  • Glycopolypeptides such as immunoglobulins, ovalbumin, lipase, glucocerebrosidase, lectins, tissue plasminogen activator and glycosylated interleukins, interferons and colony stimulating factors are of interest, as are immunoglobulins such as IgG, IgE, IgM, IgA, IgD and fragments thereof.
  • immunoglobulins such as IgG, IgE, IgM, IgA, IgD and fragments thereof.
  • receptor and ligand binding antibody-like proteins and antibodies and antibody fragments which are used in clinical medicine for diagnostic and therapeutic purposes.
  • the antibody-like protein herein is preferred (A) : the group consisting of an antibody, a antibody-like protein molecule, probody, nanobody, peptides, an antibody coating on polymeric micelle, an antibody-liposome, a lipoprotein-based drug carrier, an antibody coating on nano-particle, an antibody-dendrimer, and a particle said above coated or linked with an antibody-like protein (antibody) , or a combination of said above thereof;
  • (B) an antibody-like protein, full-length antibodies (polyclonal antibodies, monoclonal antibodies, dimers, multimers, multispecific antibodies (e.g., bispecific antibody, trispecific antibody, or tetraspecific antibody) ; single chain antibodies; an antibody fragment that binds to the target cell, a monoclonal antibody, a single chain monoclonal antibody, a monoclonal antibody fragment that binds the target cell, a chimeric antibody, a chimeric antibody fragment that binds to the target cell, a domain antibody, a domain antibody fragment that binds to the target cell, a resurfaced antibody, a resurfaced single chain antibody, or a resurfaced antibody fragment that binds to the target cell, a humanized antibody or a resurfaced antibody, a humanized single chain antibody, or a humanized antibody fragment that binds to the target cell, anti-idiotypic (anti-Id) antibodies, CDR's , diabody, triabody,
  • the fragments of antibodies include Fab, Fab', F (ab') 2 , F v , [Parham, J. Immunol. 131, 2895-902 (1983) ] , fragments produced by a Fab expression library, and epitope-binding fragments of any of the above which immuno-specifically bind to cancer cell antigens, viral antigens, microbial antigens or an antibody-like protein generated by the immune system that is capable of recognizing, binding to a specific antigen or exhibiting the desired biological activity (Miller et al (2003) J.
  • interferons such as type I, II, III
  • peptides such as IL-2, IL-3, IL-4, IL-5, IL-6, IL-10, GM-CSF, interferon-gamma (IFN- ⁇ )
  • hormones such as insulin, TRH (thyrotropin releasing hormones) , MSH (melanocyte-stimulating hormone) , steroid hormones, such as androgens and estrogens, melanocyte-stimulating hormone (MSH)
  • growth factors and colony-stimulating factors such as epidermal growth factors (EGF) , granulocyte-macrophage colony-stimulating factor (GM-CSF) , transforming growth factors (TGF) , such as TGF ⁇ , TGF ⁇ , insulin and insulin like growth factors (IGF-I, IGF-II) G-CSF, M-CSF and GM-CSF [Burgess,
  • bioactive polymers Dhar, et al, Proc. Natl. Acad. Sci. 2008, 105, 17356-61
  • bioactive dendrimers Lee, et al, Nat. Biotechnol. 2005, 23, 15
  • a monoclonal antibody is preferred as a cell-surface binding agent if an appropriate one is available.
  • the antibody may be murine, human, humanized, chimeric, or derived from other species.
  • Production of antibodies used in the present invention involves in vivo or in vitro procedures or combinations thereof.
  • Methods for producing polyclonal anti-receptor peptide antibodies are well-known in the art, such as in U.S. Pat. No. 4,493,795 (to Nestor et al) .
  • a monoclonal antibody is typically made by fusing myeloma cells with the spleen cells from a mouse that has been immunized with the desired antigen ( G.; Milstein, C. (1975) . Nature 256: 495-7) .
  • the detailed procedures are described in “Antibodies--A Laboratory Manual” , Harlow and Lane, eds., Cold Spring Harbor Laboratory Press, New York (1988) , which is incorporated herein by reference.
  • Particularly monoclonal antibodies are produced by immunizing mice, rats, hamsters or any other mammal with the antigen of interest such as the intact target cell, antigens isolated from the target cell, whole virus, attenuated whole virus, and viral proteins.
  • Splenocytes are typically fused with myeloma cells using polyethylene glycol (PEG) 6000.
  • Fused hybrids are selected by their sensitivity to HAT (hypoxanthine-aminopterin-thymine) .
  • Hybridomas producing a monoclonal antibody useful in practicing this invention are identified by their ability to immunoreact specified receptors or inhibit receptor activity on target cells.
  • a monoclonal antibody used in the present invention can be produced by initiating a monoclonal hybridoma culture comprising a nutrient medium containing a hybridoma that secretes antibody molecules of the appropriate antigen specificity.
  • the culture is maintained under conditions and for a time period sufficient for the hybridoma to secrete the antibody molecules into the medium.
  • the antibody-containing medium is then collected.
  • the antibody molecules can then be further isolated by well-known techniques, such as using protein-A affinity chromatography; anion, cation, hydrophobic, or size exclusive chromatographies (particularly by affinity for the specific antigen after protein A, and sizing column chromatography) ; centrifugation, differential solubility, or by any other standard technique for the purification of proteins.
  • Dulbecco s minimal essential medium (DMEM; Dulbecco et al., Virol. 8, 396 (1959) ) supplemented with 4.5 gm/l glucose, 0 ⁇ 20 mM glutamine, 0 ⁇ 20%fetal calf serum, several ppm amount of heavy metals, such as Cu, Mn, Fe, or Zn, etc, or/and the other heavy metals added in their salt forms, and with an anti-foaming agent, such as polyoxyethylene-polyoxypropylene block copolymer.
  • antibody-producing cell lines can also be created by techniques other than fusion, such as direct transformation of B lymphocytes with oncogenic DNA, or transfection with an oncovirus, such as Epstein-Barr virus (EBV, also called human herpesvirus 4 (HHV-4) ) or Kaposi’s sarcoma-associated herpesvirus (KSHV) .
  • EBV Epstein-Barr virus
  • HHV-4 human herpesvirus 4
  • KSHV Kaposi’s sarcoma-associated herpesvirus
  • a monoclonal antibody may also be produced via an anti-receptor peptide or peptides containing the carboxyl terminal as described well-known in the art. See Niman et al., Proc. Natl. Acad. Sci. USA, 80: 4949-53 (1983) ; Geysen et al., Proc. Natl. Acad. Sci. USA, 82: 178-82 (1985) ; Lei et al. Biochemistry 34 (20) : 6675-88, (1995) .
  • the anti-receptor peptide or a peptide analog is used either alone or conjugated to an immunogenic carrier, as the immunogen for producing anti-receptor peptide monoclonal antibodies.
  • phage display technology which can be used to select a range of human antibodies binding specifically to the antigen using methods of affinity enrichment. Phage display has been thoroughly described in the literature and the construction and screening of phage display libraries are well known in the art, see, e.g., Dente et al, Gene. 148 (1) : 7-13 (1994) ; Little et al, Biotechnol Adv. 12 (3) : 539-55 (1994) ; Clackson et al., Nature 352: 264-8 (1991) ; Huse et al., Science 246: 1275-81 (1989) .
  • Monoclonal antibodies derived by hybridoma technique from another species than human, such as mouse, can be humanized to avoid human anti-mouse antibodies when infused into humans.
  • complementarity-determining region grafting and resurfacing are more common methods of humanization of antibodies. These methods have been extensively described, see e.g. U.S. Pat. Nos. 5,859,205 and 6,797,492; Liu et al, Immunol Rev. 222: 9-27 (2008) ; Almagro et al, Front Biosci. 13: 1619-33 (2008) ; Lazar et al, Mol Immunol. 44 (8) : 1986-98 (2007) ; Li et al, Proc. Natl. Acad. Sci. U S A.
  • Fully human antibodies can also be prepared by immunizing transgenic mice, rabbits, monkeys, or other mammals, carrying large portions of the human immunoglobulin heavy and light chains, with an immunogen. Examples of such mice are: the Xenomouse. (Abgenix/Amgen) , the HuMAb-Mouse (Medarex/BMS) , the VelociMouse (Regeneron) , see also U.S. Pat. Nos. 6,596,541, 6,207,418, 6,150,584, 6,111,166, 6,075,181, 5,922,545, 5,661,016, 5,545,806, 5,436,149 and 5,569,825.
  • murine variable regions and human constant regions can also be fused to construct called “chimeric antibodies” that are considerably less immunogenic in man than murine mAbs (Kipriyanov et al, Mol Biotechnol. 26: 39-60 (2004) ; Houdebine, Curr Opin Biotechnol. 13: 625-9 (2002) each incorporated herein by reference) .
  • site-directed mutagenesis in the variable region of an antibody can result in an antibody with higher affinity and specificity for its antigen (Brannigan et al, Nat Rev Mol Cell Biol. 3: 964-70, (2002) ) ; Adams et al, J Immunol Methods. 231: 249-60 (1999) ) and exchanging constant regions of a mAb can improve its ability to mediate effector functions of binding and cytotoxicity.
  • Antibodies immunospecific for a malignant cell antigen can also be obtained commercially or produced by any method known to one of skill in the art such as, e.g., chemical synthesis or recombinant expression techniques.
  • the nucleotide sequence encoding antibodies immune-specific for a malignant cell antigen can be obtained commercially, e.g., from the GenBank database or a database like it, the literature publications, or by routine cloning and sequencing.
  • an antibody like peptide or protein that bind/block/target or in some other way interact with the epitopes or corresponding receptors on a targeted cell can be used as a binding molecule.
  • These antibody like peptides or proteins could be any random peptide or proteins that have an affinity for the epitopes or corresponding receptors and they don't necessarily have to be of the immune-globulin family.
  • These peptides can be isolated by similar techniques as for phage display antibodies (Szardenings, J Recept Signal Transduct Res. 2003, 23 (4) : 307-49) .
  • the use of peptides from such random peptide libraries can be similar to antibodies and antibody fragments.
  • binding molecules of antibody like peptides or proteins may be conjugated on or linked to a large molecules or materials, such as, but is not limited, an albumin, a polymer, a liposome, a nano particle, a dendrimer, as long as such attachment permits the peptide or protein to retain its antigen binding specificity.
  • a large molecules or materials such as, but is not limited, an albumin, a polymer, a liposome, a nano particle, a dendrimer, as long as such attachment permits the peptide or protein to retain its antigen binding specificity.
  • antibodies used for conjugation of drugs via the linkers of this prevention for treating cancer, autoimmune disease, and/or infectious disease include, but are not limited to, 3F8 (anti-GD2) , Abagovomab (anti CA-125) , Abciximab (anti CD41 (integrin alpha-IIb) , Adalimumab (anti-TNF- ⁇ ) , Adecatumumab (anti-EpCAM, CD326) , Afelimomab (anti-TNF- ⁇ ) ; Afutuzumab (anti-CD20) , Alacizumab pegol (anti-VEGFR2) , ALD518 (anti-IL-6) , Alemtuzumab (Campath, MabCampath, anti-CD52) , Altumomab (anti-CEA) , Anatumomab (anti-TAG-72) , Anrukinzumab (IMA-638, anti-IL-13) , Apolio
  • Avicidin for Breast, Colon and Rectal cancers
  • anti-EPCAM epidermal cell adhesion molecule
  • anti-TACSTD1 Tumor-associated calcium signal transducer 1
  • anti-GA733-2 gastrointestinal tumor-associated protein 2
  • anti-EGP-2 epidermal glycoprotein 2
  • anti-KSA KS1/4 antigen
  • M4S tumor antigen 17-1A
  • LymphoCide Immunomedics, NJ
  • Smart ID10 Protein Design Labs
  • Oncolym Techniclone Inc, CA
  • Allomune BioTransplant, CA
  • anti-VEGF Geneentech, CA
  • CEAcide Immunomedics, NJ
  • IMC-1C11 ImClone, NJ
  • Cetuximab ImClone, NJ
  • antibodies as cell binding molecules/ligands include, but are not limited to, are antibodies against the following antigens: Aminopeptidase N (CD13) , Annexin A1, B7-H3 (CD276, various cancers) , CA125 (ovarian) , CA15-3 (carcinomas) , CA19-9 (carcinomas) , L6 (carcinomas) , Lewis Y (carcinomas) , Lewis X (carcinomas) , alpha fetoprotein (carcinomas) , CA242 (colorectal) , placental alkaline phosphatase (carcinomas) , prostate specific antigen (prostate) , prostatic acid phosphatase (prostate) , epidermal growth factor (carcinomas) , CD2 (Hodgkin’s disease, NHL lymphoma, multiple myeloma) , CD3 epsilon (T cell lymphoma, lung, breast, gastric, ovarian cancers, autoimmune diseases
  • the antibody-like protein more preferred an IgG antibody that is able to against tumor cells, virus infected cells, microorganism infected cells, parasite infected cells, autoimmune disease cells, activated tumor cells, myeloid cells, activated T-cells, an affecting B cells, or melanocytes.
  • the antibody is able to against abnormal cells expressing any one of the following antigens or receptors: CD1, CD1a, CD1b, CD1c, CD1d, CD1e, CD2, CD3, CD3d, CD3e, CD3g, CD4, CD5, CD6, CD7, CD8, CD8a, CD8b, CD9, CD10, CD11a, CD11b, CD11c, CD11d, CD12w, CD14, CD15, CD16, CD16a, CD16b, CDw17, CD18, CD19, CD20, CD21, CD22, CD23, CD24, CD25, CD26, CD27, CD28, CD29, CD30, CD31, CD32, CD32a, CD32b, CD33, CD34, CD35, CD36, CD37, CD38, CD39, CD40, CD41, CD42, CD42a, CD42b, CD42c, CD42d, CD43, CD44, CD45, CD46, CD47, CD48, CD49b, CD49c, CD3,
  • coli shiga toxin type-1 E. coli shiga toxin type-2, ED-B, EGFL7 (EGF-like domain-containing protein 7) , EGFR, EGFRII, EGFRvIII, Endoglin, Endothelin B receptor, Endotoxin, EpCAM (epithelial cell adhesion molecule) , EphA2, Episialin, ERBB2 (Epidermal Growth Factor Receptor 2) , ERBB3, ERG (TMPRSS2 ETS fusion gene) , Escherichia coli, ETV6-AML, FAP (Fibroblast activation protein alpha) , FCGR1, alpha-Fetoprotein, Fibrin II, beta chain, Fibronectin extra domain-B, FOLR (folate receptor) , Folate receptor alpha, Folate hydrolase, Fos-related antigen 1F protein of respiratory syncytial virus, Frizzled receptor, Fucosyl GM1, GD2 ganglio
  • the antibody-drug conjugates of this invention are used for the targeted treatment of cancers.
  • the targeted cancers include, but are not limited, Adrenocortical Carcinoma, Anal Cancer, Bladder Cancer, Brain Tumor (Adult, Brain Stem Glioma, Childhood, Cerebellar Astrocytoma, Cerebral Astrocytoma, Ependymoma, Medulloblastoma, Supratentorial Primitive Neuroectodermal and Pineal Tumors, Visual Pathway and Hypothalamic Glioma) , Breast Cancer, Carcinoid Tumor, Gastrointestinal, Carcinoma of Unknown Primary, Cervical Cancer, Colon Cancer, Endometrial Cancer, Esophageal Cancer, Extrahepatic Bile Duct Cancer, Ewings Family of Tumors (PNET) , Extracranial Germ Cell Tumor, Eye Cancer, Intraocular Melanoma, Gallbladder Cancer, Gastric Cancer (Stomach) ,
  • the antibody-drug conjugates of this invention are used in accordance with the compositions and methods for the treatment or prevention of an autoimmune disease.
  • the autoimmune diseases include, but are not limited, Achlorhydra Autoimmune Active Chronic Hepatitis, Acute Disseminated Encephalomyelitis, Acute hemorrhagic leukoencephalitis, Addison’s Disease, Agammaglobulinemia, Alopecia areata, Amyotrophic Lateral Sclerosis, Ankylosing Spondylitis, Anti-GBM/TBM Nephritis, Antiphospholipid syndrome, Antisynthetase syndrome, Arthritis, Atopic allergy, Atopic Dermatitis, Autoimmune Aplastic Anemia, Autoimmune cardiomyopathy, Autoimmune hemolytic anemia, Autoimmune hepatitis, Autoimmune inner ear disease, Autoimmune lymphoproliferative syndrome, Autoimmune peripheral neuropathy, Autoimmune pancreatitis, Auto
  • the antibody-drug conjugates of this invention for the treatment or prevention of an autoimmune disease can be, but are not limited to, anti-elastin antibody; Abys against epithelial cells antibody; Anti-Basement Membrane Collagen Type IV Protein antibody; Anti-Nuclear Antibody; Anti ds DNA; Anti ss DNA, Anti Cardiolipin Antibody IgM, IgG; anti-celiac antibody; Anti Phospholipid Antibody IgK, IgG; Anti SM Antibody; Anti Mitochondrial Antibody; Thyroid Antibody; Microsomal Antibody, T-cells antibody; Thyroglobulin Antibody, Anti SCL-70; Anti-Jo; Anti-U. sub.
  • the binding molecule for the conjugate in the present invention can bind to both a receptor and a receptor complex expressed on an activated lymphocyte which is associated with an autoimmune disease.
  • the receptor or receptor complex can comprise an immunoglobulin gene superfamily member (e.g. CD2, CD3, CD4, CD8, CD19, CD20, CD22, CD28, CD30, CD33, CD37, CD38, CD56, CD70, CD79, CD79b, CD90, CD125, CD137, CD138, CD147, CD152/CTLA-4, PD-1, or ICOS) , a TNF receptor superfamily member (e.g.
  • useful cell binding ligands that are immunospecific for a viral or a microbial antigen are humanized or human monoclonal antibodies.
  • viral antigen includes, but is not limited to, any viral peptide, polypeptide protein (e.g. HIV gp120, HIV nef, RSV F glycoprotein, influenza virus neuramimi-dase, influenza virus hemagglutinin, HTLV tax, herpes simplex virus glycoprotein (e.g. gB, gC, gD, and gE) and hepatitis B surface antigen) that is capable of eliciting an immune response.
  • polypeptide protein e.g. HIV gp120, HIV nef, RSV F glycoprotein, influenza virus neuramimi-dase, influenza virus hemagglutinin, HTLV tax, herpes simplex virus glycoprotein (e.g. gB, gC, gD, and gE) and hepatitis B surface antigen
  • microbial antigen includes, but is not limited to, any microbial peptide, polypeptide, protein, saccharide, polysaccharide, or lipid molecule (e.g., bacteria, fungi, pathogenic protozoa, or yeast polypeptides including, e.g., LPS and capsular polysaccharide 5/8) that is capable of eliciting an immune response.
  • microbial antigen includes, but is not limited to, any microbial peptide, polypeptide, protein, saccharide, polysaccharide, or lipid molecule (e.g., bacteria, fungi, pathogenic protozoa, or yeast polypeptides including, e.g., LPS and capsular polysaccharide 5/8) that is capable of eliciting an immune response.
  • antibodies available l for the viral or microbial infection include, but are not limited to, Palivizumab which is a humanized anti-respiratory syncytial virus monoclonal antibody for the treatment of RSV infection; PRO542 which is a CD4 fusion antibody for the treatment of HIV infection; Ostavir which is a human antibody for the treatment of hepatitis B virus; PROTVIR which is a humanized IgG. sub. 1 antibody for the treatment of cytomegalovirus; and anti-LPS antibodies.
  • Palivizumab which is a humanized anti-respiratory syncytial virus monoclonal antibody for the treatment of RSV infection
  • PRO542 which is a CD4 fusion antibody for the treatment of HIV infection
  • Ostavir which is a human antibody for the treatment of hepatitis B virus
  • PROTVIR which is a humanized IgG. sub. 1 antibody for the treatment of cytomegalovirus
  • anti-LPS antibodies include, but are not limited to,
  • infectious diseases include, but are not limited to, Acinetobacter infections, Actinomycosis, African sleeping sickness (African trypanosomiasis) , AIDS (Acquired immune deficiency syndrome) , Amebiasis, Anaplasmosis, Anthrax, Arcano-bacterium haemolyticum infection, Argentine hemorrhagic fever, Ascariasis, Aspergillosis, Astrovirus infection, Babesiosis, Bacillus cereus infection, Bacterial pneumonia, Bacterial vaginosis, Bacteroides infection, Balantidiasis, Baylisascaris infection, BK virus infection, Black piedra, Blastocystis hominis infection, Blastomycosis, Venezuelan hemorrhagic fever, Borrelia infection, Botulism (and Infant botulism) , Brazilian hemorrhagic fever, Brucellosis
  • the cell binding molecule which is more preferred to be an antibody described in this patent that are against pathogenic strains include, but are not limit, Acinetobacter baumannii, Actinomyces israelii, Actinomyces gerencseriae and Propionibacterium propionicus, Trypanosoma brucei, HIV (Human immunodeficiency virus) , Entamoeba histolytica, Anaplasma genus, Bacillus anthracis, Arcanobacterium haemolyticum, Junin virus, Ascaris lumbricoides, Aspergillus genus, Astroviridae family, Babesia genus, Bacillus cereus, multiple bacteria, Bacteroides genus, Balantidium coli, Baylisascaris genus, BK virus, Piedraia hortae, Blastocystis hominis, Blastomyces dermatitides, Machupo virus, Borrelia genus, Clostri
  • antibodies as cell binding ligands used in this invention for treatment of viral disease include, but are not limited to, antibodies against antigens of pathogenic viruses, including as examples and not by limitation: Poxyiridae, Herpesviridae, Adenoviridae, Papovaviridae, Enteroviridae, Picornaviridae, Parvoviridae, Reoviridae, Retroviridae, influenza viruses, parainfluenza viruses, mumps, measles, respiratory syncytial virus, rubella, Arboviridae, Rhabdoviridae, Arenaviridae, Non-A/Non-B Hepatitis virus, Rhinoviridae, Coronaviridae, Rotoviridae, Oncovirus [such as, HBV (Hepatocellular carcinoma) , HPV (Cervical cancer, Anal cancer) , Kaposi’s sarcoma-associated herpesvirus (Kaposi’s sarcoma) , Epstein-Bar
  • the present invention also concerns pharmaceutical compositions comprising the conjugate of the invention together with a pharmaceutically acceptable carrier, diluent, or excipient for treatment of cancers, infections or autoimmune disorders.
  • a pharmaceutically acceptable carrier diluent, or excipient for treatment of cancers, infections or autoimmune disorders.
  • the method for treatment of cancers, infections and autoimmune disorders can be practiced in vitro, in vivo, or ex vivo.
  • in vitro uses include treatments of cell cultures in order to kill all cells except for desired variants that do not express the target antigen; or to kill variants that express undesired antigen.
  • ex vivo uses include treatments of hematopoietic stem cells (HSC) prior to the performance of the transplantation (HSCT) into the same patient in order to kill diseased or malignant cells.
  • HSC hematopoietic stem cells
  • the bone marrow cells are washed with medium containing serum and returned to the patient by i.v. infusion according to known methods.
  • the treated marrow cells are stored frozen in liquid nitrogen using standard medical equipment.
  • the conjugates of the patent application are formulated to liquid, or suitable to be lyophilized and subsequently be reconstituted to a liquid formulation.
  • the conjugate in a liquid formula or in the formulated lyophilized powder may take up 0.01%-99%by weight as major gradient in the formulation.
  • a liquid formulation comprising 0.1 g/L ⁇ 300 g/L of concentration of the conjugate active ingredient for delivery to a patient without high levels of antibody aggregation may include one or more polyols (e.g. sugars) , a buffering agent with pH 4.5 to 7.5, a surfactant (e.g. polysorbate 20 or 80) , an antioxidant (e.g.
  • a tonicity agent e.g. mannitol, sorbitol or NaCl
  • chelating agents such as EDTA
  • metal complexes e.g. Zn-protein complexes
  • biodegradable polymers such as polyesters
  • a preservative e.g. benzyl alcohol
  • Suitable buffering agents for use in the formulations include, but are not limited to, organic acid salts such as sodium, potassium, ammounium, or trihydroxyethylamino salts of citric acid, ascorbic acid, gluconic acid, carbonic acid, tartaric acid, succinic acid, acetic acid or phtalic acid; Tris, tromethamine hydrochloride, sulfate or phosphate buffer.
  • amino acid cationic components can also be used as buffering agent.
  • amino acid component includes without limitation arginine, glycine, glycylglycine, and histidine.
  • the arginine buffers include arginine acetate, arginine chloride, arginine phosphate, arginine sulfate, arginine succinate, etc.
  • the arginine buffer is arginine acetate.
  • histidine buffers include histidine chloride-arginine chloride, histidine acetate-arginine acetate, histidine phosphate-arginine phosphate, histidine sulfate-arginine sulfate, histidine succinate-argine succinate, etc.
  • the formulations of the buffers have a pH of 4.5 to pH 7.5, preferably from about 4.5 to about 6.5, more preferably from about 5.0 to about 6.2.
  • the concentration of the organic acid salts in the buffer is from about 10 mM to about 500 mM.
  • a “polyol” that may optionally be included in the formulation is a substance with multiple hydroxyl groups.
  • Polyols can be used as stabilizing excipients and/or isotonicity agents in both liquid and lyophilized formulations.
  • Polyols can protect biopharmaceuticals from both physical and chemical degradation pathways.
  • Preferentially excluded co-solvents increase the effective surface tension of solvent at the protein interface whereby the most energetically favorable structural conformations are those with the smallest surface areas.
  • Polyols include sugars (reducing and nonreducing sugars) , sugar alcohols and sugar acids.
  • a “reducing sugar” is one which contains a hemiacetal group that can reduce metal ions or react covalently with lysine and other amino groups in proteins and a “nonreducing sugar” is one which does not have these properties of a reducing sugar.
  • reducing sugars are fructose, mannose, maltose, lactose, arabinose, xylose, ribose, rhamnose, galactose and glucose.
  • Nonreducing sugars include sucrose, trehalose, sorbose, melezitose and raffinose.
  • Sugar alcohols are selected from mannitol, xylitol, erythritol, maltitol, lactitol, erythritol, threitol, sorbitol and glycerol.
  • Sugar acids include L-gluconate and metallic salts thereof.
  • the polyol in the liquid formula or in the formulated lyophilized solid can be 0.0%-20%by weight.
  • a nonreducing sugar, sucrose or trehalose at a concentration of about from 0.1%to 15% is chosen in the formulation, wherein trehalose being preferred over sucrose, because of the solution stability of trehalose.
  • a surfactant optionally in the formulations is selected from polysorbate (polysorbate 20, polysorbate 40, polysorbate 65, polysorbate 80, polysorbate 81, polysorbate 85 and the like) ; poloxamer (e.g. poloxamer 188, poly (ethylene oxide) -poly (propylene oxide) , poloxamer 407 or polyethylene-polypropylene glycol and the like) ; Triton; sodium dodecyl sulfate (SDS) ; sodium laurel sulfate; sodium octyl glycoside; lauryl-, myristyl-, linoleyl-, or stearyl-sulfobetaine; lauryl-, myristyl-, linoleyl-or stearyl-sarcosine; linoleyl-, myristyl-, or cetyl-betaine; lauroamidopropyl-, cocamidopropy
  • lauroamidopropyl myristamidopropyl-, palmidopropyl-, or isostearamido-propyl-dimethylamine; sodium methyl cocoyl-, or disodium methyl oleyl-taurate; dodecyl betaine, dodecyl dimethylamine oxide, cocamidopropyl betaine and coco ampho glycinate; and the MONAQUAT TM series (e.g. isostearyl ethylimidonium ethosulfate) ; polyethyl glycol, polypropyl glycol, and copolymers of ethylene and propylene glycol (e.g.
  • Preferred surfactants are polyoxyethylene sorbitan fatty acid esters e.g. polysorbate 20, 40, 60 or 80 (Tween 20, 40, 60 or 80) .
  • the concentration of a surfactant in the formulation is range from 0.0%to about 2.0%by weight. In certain embodiments, the surfactant concentration is from about 0.01%to about 0.2%. In one embodiment, the surfactant concentration is about 0.02%.
  • a “preservative” optionally in the formulations is a compound that essentially reduces bacterial action therein.
  • potential preservatives include octadecyldimethylbenzyl ammonium chloride, hexamethonium chloride, benzalkonium chloride (amixture of alkylbenzyldimethylammonium chlorides in which the alkyl groups are long-chain compounds) , and benzethonium chloride.
  • preservatives include aromatic alcohols such as phenoxyl, butyl and benzyl alcohol, alkyl parabens such as methyl or propyl paraben, catechol, resorcinol, cyclohexanol, 3-pentanol, and m-cresol.
  • aromatic alcohols such as phenoxyl, butyl and benzyl alcohol
  • alkyl parabens such as methyl or propyl paraben
  • catechol resorcinol
  • cyclohexanol 3-pentanol
  • m-cresol m-cresol
  • the preservative in the liquid formula or in the formulated lyophilized powder can be 0.0%-5.0%by weight.
  • the preservative herein is benzyl alcohol.
  • Suitable free amino acids as a bulky material, or tonicity agent, or osmotic pressure adjustment in the formulation is selected from, but are not limited to, one or more of arginine, cystine, glycine, lysine, histidine, ornithine, isoleucine, leucine, alanine, glycine glutamic acid or aspartic acid.
  • arginine, cystine, glycine, lysine, histidine, ornithine isoleucine, leucine, alanine, glycine glutamic acid or aspartic acid.
  • the inclusion of a basic amino acid is preferred i.e. arginine, lysine and/or histidine. If a composition includes histidine then this may act both as a buffering agent and a free amino acid, but when a histidine buffer is used it is typical to include a non-histidine free amino acid e.g.
  • amino acid may be present in its D-and/or L-form, but the L-form is typical.
  • the amino acid may be present as any suitable salt e.g. a hydrochloride salt, such as arginine-HCl.
  • the amino acid in the liquid formula or in the formulated lyophilized powder can be 0.0%-30%by weight.
  • the formulations can optionally comprise methionine, glutathione, cysteine, cystine or ascorbic acid as an antioxidant at a concentration of about up to 5 mg/ml in the liquid formula or 0.0%-5.0%by weight in the formulated lyophilized powder;
  • the formulations can optionally comprise metal chelating agent, e.g., EDTA, EGTA, etc., at a concentration of about up to 2 mM in the liquid formula or 0.0%-0.3%by weight in the formulated lyophilized powder.
  • the final formulation can be adjusted to the preferred pH with a buffer adjusting agent (e.g. an acid, such as HCl, H 2 SO 4 , acetic acid, H 3 PO 4 , citric acid, etc, or a base, such as NaOH, KOH, NH 4 OH, ethanolamine, diethanolamine or triethanol amine, sodium phosphate, potassium phosphate, trisodium citrate, tromethamine, etc) and the formulation should be controlled “isotonic” which is meant that the formulation of interest has essentially the same osmotic pressure as human blood. Isotonic formulations will generally have an osmotic pressure from about 250 to 350 mOsm.
  • a buffer adjusting agent e.g. an acid, such as HCl, H 2 SO 4 , acetic acid, H 3 PO 4 , citric acid, etc, or a base, such as NaOH, KOH, NH 4 OH, ethanolamine, diethanolamine or triethanol amine, sodium phosphat
  • Isotonicity can be measured using a vapor pressure or ice-freezing type osmometer, for example.
  • the isotonic agent is selected from mannitol, sorbitol, sodium acetate, potassium chloride, sodium phosphate, potassium phosphate, trisodium citrate, or NaCl.
  • both the buffer salts and the isotonic agent may take up to 30%by weight in the formulation.
  • excipients which may be useful in either a liquid or lyophilized formulation of the patent application include, for example, fucose, cellobiose, maltotriose, melibiose, octulose, ribose, xylitol, arginine, histidine, glycine, alanine, methionine, glutamic acid, lysine, imidazole, glycylglycine, mannosylglycerate, Triton X-100, Pluoronic F-127, cellulose, cyclodextrin, (2-Hydroxypropyl) - ⁇ -cyclodextrin, dextran (10, 40 and/or 70 kD) , polydextrose, maltodextrin, ficoll, gelatin, hydroxypropylmeth, sodium phosphate, potassium phosphate, ZnCl 2 , zinc, zinc oxide, sodium citrate, trisodium citrate
  • contemplated excipients which may be utilized in the aqueous pharmaceutical compositions of the patent application include, for example, flavoring agents, antimicrobial agents, sweeteners, antioxidants, antistatic agents, lipids such as phospholipids or fatty acids, steroids such as cholesterol, protein excipients such as serum albumin (human serum albumin) , recombinant human albumin, gelatin, casein, salt-forming counterions such sodium and the like.
  • a pharmaceutical container or vessel is used to hold the pharmaceutical formulation of any of conjugates of the patent application.
  • the vessel is a vial, bottle, pre-filled syringe, pre-filled or auto-injector syringe.
  • the liquid formula can be freeze-dried or drum-dryed to a form of cake or powder in a borosilicate vial or soda lime glass vial.
  • the solid powder can also be prepared by efficient spray drying, and then packed to a vial or a pharmaceutical container for storage and distribution.
  • the invention provides a method for preparing a formulation comprising the steps of: (a) lyophilizing the formulation comprising the conjugates, excipients, and a buffer system; and (b) reconstituting the lyophilized mixture of step (a) in a reconstitution medium such that the reconstituted formulation is stable.
  • the formulation of step (a) may further comprise a stabilizer and one or more excipients selected from a group comprising bulking agent, salt, surfactant and preservative as hereinabove described.
  • reconstitution media several diluted organic acids or water, i.e. sterile water, bacteriostatic water for injection (BWFI) or may be used.
  • the reconstitution medium may be selected from water, i.e.
  • sterile water bacteriostatic water for injection (BWFI) or the group consisting of acetic acid, propionic acid, succinic acid, sodium chloride, magnesium chloride, acidic solution of sodium chloride, acidic solution of magnesium chloride and acidic solution of arginine, in an amount from about 10 to about 250 mM.
  • BWFI bacteriostatic water for injection
  • a liquid pharmaceutical formulation of the conjugates of the patent application should exhibit a variety of pre-defined characteristics.
  • One of the major concerns in liquid drug products is stability, as proteins/antibodies tend to form soluble and insoluble aggregates during manufacturing and storage.
  • various chemical reactions can occur in solution (deamidation, oxidation, clipping, isomerization etc. ) leading to an increase in degradation product levels and/or loss of bioactivity.
  • a conjugate in either liquid or loyphilizate formulation should exhibit a shelf life of more than 6 months at 25°C. More preferred a conjugate in either liquid or loyphilizate formulation should exhibit a shelf life of more than 12 months at 25°C.
  • liquid formulation should exhibit a shelf life of about 24 to 36 months at 2-8°C and the loyphilizate formulation should exhibit a shelf life of about preferably up to 60 months at 2-8°C. Both liquid and loyphilizate formulations should exhibit a shelf life for at least two years at -20°C, or -70°C.
  • the formulation is stable following freezing (e.g., -20°C, or -70°C. ) and thawing of the formulation, for example following 1, 2 or 3 cycles of freezing and thawing.
  • Stability can be evaluated qualitatively and/or quantitatively in a variety of different ways, including evaluation of drug/antibody (protein) ratio and aggregate formation (for example using UV, size exclusion chromatography, by measuring turbidity, and/or by visual inspection) ; by assessing charge heterogeneity using cation exchange chromatography, image capillary isoelectric focusing (icIEF) or capillary zone electrophoresis; amino-terminal or carboxy-terminal sequence analysis; mass spectrometric analysis, or matrix-assisted laser desorption ionization/time-of-flight mass spectrometry (MALDI/TOF MS) , or HPLC-MS/MS; SDS-PAGE analysis to compare reduced and intact antibody; peptide map (for example tryptic or LYS--C
  • Instability may involve any one or more of: aggregation, deamidation (e.g. Asn deamidation) , oxidation (e.g. Met oxidation) , isomerization (e.g. Asp isomeriation) , clipping/hydrolysis/fragmentation (e.g. hinge region fragmentation) , succinimide formation, unpaired cysteine (s) , N-terminal extension, C-terminal processing, glycosylation differences, etc.
  • deamidation e.g. Asn deamidation
  • oxidation e.g. Met oxidation
  • isomerization e.g. Asp isomeriation
  • clipping/hydrolysis/fragmentation e.g. hinge region fragmentation
  • a stable conjugate should also “retains its biological activity” in a pharmaceutical formulation, if the biological activity of the conjugate at a given time, e.g. 24 month, within about 20%, preferably about 10% (within the errors of the assay) of the biological activity exhibited at the time the pharmaceutical formulation was prepared as determined in an antigen binding assay, and/or in vitro, cytotoxic assay, for example.
  • the conjugate of the invention will be supplied as solutions or as a lyophilized solid that can be redissolved in sterile water for injection.
  • suitable protocols of conjugate administration are as follows. Conjugates are given dayly, weekly, biweekly, triweekly, once every four weeks or monthly for 8 ⁇ 54 weeks as an i.v. bolus. Bolus doses are given in 50 to 1000 ml of normal saline to which human serum albumin (e.g. 0.5 to 1 mL of a concentrated solution of human serum albumin, 100 mg/mL) can optionally be added. Dosages will be about 50 ⁇ g to 20 mg/kg of body weight per week, i.v.
  • Examples of medical conditions that can be treated according to the in vivo or ex vivo methods of killing selected cell populations include malignancy of any types of cancer, autoimmune diseases, graft rejections, and infections (viral, bacterial or parasite) .
  • the amount of a conjugate which is required to achieve the desired biological effect will vary depending upon a number of factors, including the chemical characteristics, the potency, and the bioavailability of the conjugates, the type of disease, the species to which the patient belongs, the diseased state of the patient, the route of administration, all factors which dictate the required dose amounts, delivery and regimen to be administered.
  • the conjugates of this invention may be provided in an aqueous physiological buffer solution containing 0.1 to 10%w/v conjugates for parenteral administration.
  • Typical dose ranges are from 1 ⁇ g/kg to 0.1 g/kg of body weight daily; weekly, biweekly, triweekly, or monthly, a preferred dose range is from 0.01 mg/kg to 25 mg/kg of body weight weekly, biweekly, triweekly, or monthly, an equivalent dose in a human.
  • the preferred dosage of drug to be administered is likely to depend on such variables as the type and extent of progression of the disease or disorder, the overall health status of the particular patient, the relative biological efficacy of the compound selected, the formulation of the compound, the route of administration (intravenous, intramuscular, or other) , the pharmacokinetic properties of the conjugates by the chosen delivery route, and the speed (bolus or continuous infusion) and schedule of administrations (number of repetitions in a given period of time) .
  • the conjugates of the present invention are also capable of being administered in unit dose forms, wherein the term “unit dose” means a single dose which is capable of being administered to a patient, and which can be readily handled and packaged, remaining as a physically and chemically stable unit dose comprising either the active conjugate itself, or as a pharmaceutically acceptable composition, as described hereinafter.
  • unit doses for humans range from 1 mg to 3000 mg per day, or per week, per two weeks (biweekly) , triweekly, or per month.
  • the unit dose range is from 1 to 500 mg administered one to four times a month and even more preferably from 1 mg to 100 mg, once a week, or once a biweek, or once a triweek.
  • Conjugatess provided herein can be formulated into pharmaceutical compositions by admixture with one or more pharmaceutically acceptable excipients.
  • Such unit dose compositions may be prepared for use by oral administration, particularly in the form of tablets, simple capsules or soft gel capsules; or intranasally, particularly in the form of powders, nasal drops, or aerosols; or dermally, for example, topically in ointments, creams, lotions, gels or sprays, or via trans-dermal patches.
  • compositions may conveniently be administered in unit dosage form and may be prepared by any of the methods well known in the pharmaceutical art, for example, as described in Remington: The Science and Practice of Pharmacy, 21 th ed.; Lippincott Williams &Wilkins: Philadelphia, PA, 2005.
  • the formulations include pharmaceutical compositions in which a compound of the present invention is formulated for oral or parenteral administration.
  • tablets, pills, powders, capsules, troches and the like can contain one or more of any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, or gum tragacanth; a diluent such as starch or lactose; a disintegrant such as starch and cellulose derivatives; a lubricant such as magnesium stearate; a glidant such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or a flavoring agent such as peppermint, or methyl salicylate.
  • a binder such as microcrystalline cellulose, or gum tragacanth
  • a diluent such as starch or lactose
  • a disintegrant such as starch and cellulose derivatives
  • a lubricant such as magnesium stearate
  • a glidant such
  • Capsules can be in the form of a hard capsule or soft capsule, which are generally made from gelatin blends optionally blended with plasticizers, as well as a starch capsule.
  • dosage unit forms can contain various other materials that modify the physical form of the dosage unit, for example, coatings of sugar, shellac, or enteric agents.
  • Other oral dosage forms syrup or elixir may contain sweetening agents, preservatives, dyes, colorings, and flavorings.
  • the active compounds may be incorporated into fast dissolve, modified-release or sustained-release preparations and formulations, and wherein such sustained-release formulations are preferably bi-modal.
  • Preferred tablets contain lactose, cornstarch, magnesium silicate, croscarmellose sodium, povidone, magnesium stearate, or talc in any combination.
  • Liquid preparations for parenteral administration include sterile aqueous or non-aqueous solutions, suspensions, and emulsions.
  • the liquid compositions may also include binders, buffers, preservatives, chelating agents, sweetening, flavoring and coloring agents, and the like.
  • Non-aqueous solvents include alcohols, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and organic esters such as ethyl oleate.
  • Aqueous carriers include mixtures of alcohols and water, buffered media, and saline.
  • biocompatible, biodegradable lactide polymer, lactide/glycolide copolymer, or polyoxyethylene-polyoxypropylene copolymers may be useful excipients to control the release of the active compounds.
  • Intravenous vehicles can include fluid and nutrient replenishers, electrolyte replenishers, such as those based on Ringer’s dextrose, and the like.
  • Other potentially useful parenteral delivery systems for these active compounds include ethylene-vinyl acetate copolymer particles, osmotic pumps, implantable infusion systems, and liposomes.
  • formulations for inhalation which include such means as dry powder, aerosol, or drops. They may be aqueous solutions containing, for example, polyoxyethylene-9-lauryl ether, glycocholate and deoxycholate, or oily solutions for administration in the form of nasal drops, or as a gel to be applied intranasally.
  • Formulations for buccal administration include, for example, lozenges or pastilles and may also include a flavored base, such as sucrose or acacia, and other excipients such as glycocholate.
  • Formulations suitable for rectal administration are preferably presented as unit-dose suppositories, with a solid based carrier, such as cocoa butter, and may include a salicylate.
  • Formulations for topical application to the skin preferably take the form of an ointment, cream, lotion, paste, gel, spray, aerosol, or oil.
  • Carriers which can be used include petroleum jelly, lanolin, polyethylene glycols, alcohols, or their combinations.
  • Formulations suitable for transdermal administration can be presented as discrete patches and can be lipophilic emulsions or buffered, aqueous solutions, dissolved and/or dispersed in a polymer or an adhesive.
  • a pharmaceutical composition comprising a therapeuticcally effective amount of the conjugate of Formula (V) , (VI) , (VII) , or any conjugates described through the present patent can be administered concurrently with the other therapeutic agents such as the chemotherapeutic agent, the radiation therapy, immunotherapy agents, autoimmune disorder agents, anti-infectious agents or the other conjugates for synergistically effective treatment or prevention of a cancer, or an autoimmune disease, or an infectious disease.
  • the other therapeutic agents such as the chemotherapeutic agent, the radiation therapy, immunotherapy agents, autoimmune disorder agents, anti-infectious agents or the other conjugates for synergistically effective treatment or prevention of a cancer, or an autoimmune disease, or an infectious disease.
  • the synergistic drugs or radiation therapy can be administered prior or subsequent to administration of a conjugate, in one aspect at least an hour, 12 hours, a day, a week, biweeks, triweeks, a month, in further aspects several months, prior or subsequent to administration of a conjugate of the invention.
  • the synergistic agents are preferably selected from one or several of the following drugs:
  • the synergistic agents according to Claim 20 are selected from one or several of the following drugs: Abatacept, Abiraterone acetate, Abraxane, Acetaminophen/hydrocodone, Acalabrutinib, aducanumab, Adalimumab, ADXS31-142, ADXS-HER2, Afatinib dimaleate, Aldesleukin, Alectinib, Alemtuzumab, Alitretinoin, ado-trastuzumab emtansine, Amphetamine/dextroamphetamine, Anastrozole, Aripiprazole, anthracyclines, Aripiprazole, Atazanavir, Atezolizumab, Atorvastatin, Avelumab, Axicabtagene ciloleucel, Axitinib, Belinostat
  • the drugs/cytotoxic agents used for conjugation of the present patent can be any analogues and/or derivatives of drugs/molecules described in the present patent.
  • drugs/cytotoxic agents will readily understand that each of the drugs/cytotoxic agents described herein can be modified in such a manner that the resulting compound still retains the specificity and/or activity of the starting compound.
  • the skilled artisan will also understand that many of these compounds can be used in place of the drugs/cytotoxic agents described herein.
  • the drugs/cytotoxic agents of the present invention include analogues and derivatives of the compounds described herein.
  • the conjugate and process of the present invention may be prepared in a number of ways well known to those skilled in the art.
  • the Camptothecin analogs used in the conjugate can be synthesized, for example, by application or adaptation of the methods described below, or variations thereon as appreciated by the skilled artisan.
  • the appropriate modifications and substitutions will be readily apparent and well known or readily obtainable from the scientific literature to those skilled in the art. In particular, such methods can be found in R. C. Larock, Comprehensive Organic Transformations, 2 nd Edition, Wiley-VCH Publishers, 1999.
  • any base, acid or solvent conventionally used in reactions of this type may equally be used here, provided that it has no adverse effect on other parts of the molecule.
  • the reactions can take place over a wide range of temperatures. In general, we find it convenient to carry out the reaction at a temperature of from -80°C to 150°C (more preferably from about room temperature to 100°C) .
  • the time required for the reaction may also vary widely, depending on many factors, notably the reaction temperature and the nature of the reagents. However, provided that the reaction is effected under the preferred conditions outlined above, a period of from 3 hours to 20 hours will usually suffice.
  • reaction products may be recovered by distilling off the solvent from the reaction mixture or, if necessary after distilling off the solvent from the reaction mixture, pouring the residue into water followed by extraction with a water-immiscible organic solvent and distilling off the solvent from the extract.
  • product can, if desired, be further purified by various well known techniques, such as recrystallization, reprecipitation or the various chromatography techniques, notably column chromatography or preparative thin layer chromatography.
  • NMR spectra were recorded on Zhongke-niujin WNMR-I 400 MHz instrument at the Department of Chemistry of Zhejiang Sci-Tech University. Chemical shifts ( ⁇ ) are reported in parts per million (ppm) referenced to tetramethylsilane at 0.00 and coupling constants (J) are reported in Hz.
  • the elemental analysis of C, H, and/or N was provided by the Department of Chemistry of Zhejiang Sci-Tech University and conducted on Elementar UNICUBE. Quantitative analysis of metal atoms was performed on Agilent ICPOES 730 ICP-MS.
  • Example 2 The procedure is the same as that of Example 1, starting from zinc chloride (6.0 g, 44.03 mmol) and O-phenylenediamine (19.1 g, 176.11 mmol) to provide 9.5 g of zinc o-phenylenediamine chloride complex as an off-white solid, 88.3%yield.
  • Example 2 The procedure is the same as that of Example 1, starting from zinc chloride (4.0 g, 29.35 mmol) and pyridin-2-ylmethanamine (12.7 g, 117.41 mmol) to provide 6.2 g of zinc pyridin-2-ylmethanamine chloride complex as an off-white solid, 86.8%yield.
  • Example 2 The procedure is the same as that of Example 1, starting from zinc chloride (4.0 g, 29.35 mmol) and 4- (pyrrolidin-1-yl) pyridine (7.2 g, 58.70 mmol) to provide 8.5 g of zinc 4-dimethylaminopyridine chloride complex as an off-white solid, 66.8%yield.
  • 1 H NMR 400 MHz, DMSO-d6) ⁇ 8.07 –8.01 (m, 4H) , 6.66 –6.59 (m, 4H) , 2.02 –1.93 (m, 8H) .
  • Example 2 The procedure is the same as that of Example 1, starting from zinc chloride (4.0 g, 29.35 mmol) and 1- (pyridin-2-yl) ethan-1-amine (3.6 g, 29.35 mmol) to provide 6.6 g of zinc 1- (pyridin-2-yl) ethan-1-amine chloride complex as an off-white solid, 87.1%yield.
  • N-Boc piperidone (10 g, 0.05 mol) was dissolve in MeOH (100 mL) , to which dimethylamine aqueous solution (25 mL, 0.22 mol) and 10%palladium on carbon (1 g) were added, and the reaction flask was evacuated and re-filled with hydrogen, then stirred at r.t. overnight. After filtration, the filtrate was concentrated and co-evaporated with dichloromethane for three times (3 ⁇ 80 mL) , and dried on a vacuum pump to remove all dimethylamine. HCl/MeOH (4 M, 50 mL) was added to the residue and stirred at r.t. for 30 minutes.
  • Example 40 Synthesis of N- (4- ( (S) -2- ( (tert-butoxycarbonyl) amino) propanamido) benzyl) -1- ( ( (S) -4-ethyl-4, 9-dihydroxy-3, 14-dioxo-3, 4, 12, 14-tetrahydro-1H-pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-10-yl) methyl) -N, N-dimethylpiperidin-4-aminium bromide (7)
  • Example 51 Synthesis of 2, 5-dioxopyrrolidin-1-yl (S) -30- (4- (2, 5-dioxo-2, 5-dihydro-1H-pyrrol-1-yl) butanamido) -27, 31-dioxo-2, 5, 8, 11, 14, 17, 20, 23-octaoxa-26, 32-diazahexatriacontan-36-oate (19)
  • Example 55 Synthesis of N- (4- ( (S) -2- ( (tert-butoxycarbonyl) amino) propanamido) benzyl) -1- ( ( (S) -4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3, 14-dioxo-3, 4, 12, 14-tetrahydro-1H-pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-11-yl) methyl) -N, N-dimethylpiperidin-4-aminium chloride (23)
  • Example 56 Synthesis of N- (4- ( (S) -2-aminopropanamido) benzyl) -1- ( ( (S) -4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3, 14-dioxo-3, 4, 12, 14-tetrahydro-1H-pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-11-yl) methyl) -N, N-dimethylpiperidin-4-aminium (24)
  • Example 63 Synthesis of 4- ( (S) -30- (4- (2, 5-dioxo-2, 5-dihydro-1H-pyrrol-1-yl) butanamido) -27, 31-dioxo-2, 5, 8, 11, 14, 17, 20, 23-octaoxa-26, 32-diazahexa triacontanamido) -1- ( ( (S) -4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3, 14-dioxo-3, 4, 12, 14-tetrahydro-1H-pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-11-yl) methyl) -1-methyl piperidin-1-ium formate (31)
  • 1-methylpiperazine (5.0 g, 50.0 mmol) was dissolved in a mixed solution of 1, 4-dioxane and water (60 mL/100 mL) , and sodium bicarbonate (12.6 g, 150 mmol) was added, and the mixture was cooled to 0°C.
  • Example 68 Synthesis of 1- (4- ( (S) -2-aminopropanamido) benzyl) -4- ( ( (S) -4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3, 14-dioxo-3, 4, 12, 14-tetrahydro-1H-pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-11-yl) methyl) -1-methylpiperazin-1-ium (36)
  • Example 70 Synthesis of 1- (4- ( (30S, 38S) -30- (4- (2, 5-dioxo-2, 5-dihydro-1H-pyrrol-1-yl) butanamido) -38-methyl-27, 31, 36-trioxo-2, 5, 8, 11, 14, 17, 20, 23-octaoxa-26, 32, 37-triazanonatriacontanamido) benzyl) -4- ( ( (S) -4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3, 14-dioxo-3, 4, 12, 14-tetrahydro-1H-pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-11-yl) methyl) -1-methylpiperazin-1-ium formate (38)
  • Example 75 Synthesis of 1- (4- ( (S) -2- ( (S) -3-amino-2- (2, 5-dioxo-2, 5-dihydro-1H-pyrrol-1-yl) propanamido) propanamido) benzyl) -4- ( ( (S) -4-ethyl-8-fluoro-4-hydroxy-9-methoxy-3, 14-dioxo-3, 4, 12, 14-tetrahydro-1H-pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-11-yl) methyl) -1-methylpiperazin-1-ium (43)
  • Example 82 Synthesis of bis ( (S) -4-ethyl-4-hydroxy-3, 14-dioxo-3, 4, 12, 14-tetrahydro-1H-pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl) 4, 4'- ( ( (tert-butoxycarbonyl) azanediyl) bis (ethane-2, 1-diyl) ) bis (azanediyl) ) bis (4-oxobutanoate) (50)
  • Example 84 Synthesis of (S) - (S) -4-ethyl-4-hydroxy-3, 14-dioxo-3, 4, 12, 14-tetrahydro-1H-pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl 30- (4- (2, 5-dioxo-2, 5-dihydro-1H-pyrrol-1-yl) butanamido) -37- (2- (4- ( ( (S) -4-ethyl-4-hydroxy-3, 14-dioxo-3, 4, 12, 14-tetrahydro-1H-pyrano [3', 4': 6, 7] indolizino [1, 2-b] quinolin-9-yl) oxy) -4-oxobutanamido) ethyl) -27, 31, 36, 41-tetraoxo-2, 5, 8, 11, 14, 17, 20, 23-octaoxa-26, 32, 37, 40-tetraa
  • Iminodiacetic acid (5.0 g, 37.6 mmol) was dissolved in THF (50 mL) and water (50 mL) , mixed with NaHCO 3 (12.6 g, 150 mmol) .
  • Boc 2 O (9.8 g, 45.1 mmol) was added slowly at about 5 °C, then the reaction was warmed to r.t. and stirred for 2 days.
  • the reaction mixture was diluted with water (100 mL) , washed with ethyl acetate (2 ⁇ 30 mL) , and then adjusted to pH 1.0 using concentrated HCl.

Landscapes

  • Health & Medical Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Engineering & Computer Science (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Immunology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Cell Biology (AREA)
  • Organic Chemistry (AREA)
  • Molecular Biology (AREA)
  • Toxicology (AREA)
  • Hematology (AREA)
  • Oncology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Biophysics (AREA)
  • Genetics & Genomics (AREA)
  • Biochemistry (AREA)
  • Microbiology (AREA)
  • Mycology (AREA)
  • Peptides Or Proteins (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicinal Preparation (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
PCT/CN2021/128453 2021-11-03 2021-11-03 Specific conjugation of an antibody Ceased WO2022078524A2 (en)

Priority Applications (24)

Application Number Priority Date Filing Date Title
JP2024526535A JP2024541058A (ja) 2021-11-03 2021-11-03 抗体の特異的共役
CA3236930A CA3236930A1 (en) 2021-11-03 2021-11-03 Specific conjugation of an antibody
KR1020247017868A KR20240095442A (ko) 2021-11-03 2021-11-03 항체의 특이적 접합
EP21879565.6A EP4426727A2 (en) 2021-11-03 2021-11-03 Specific conjugation of an antibody
CN202180099689.6A CN117980327A (zh) 2021-11-03 2021-11-03 抗体的特异性偶联
PCT/CN2021/128453 WO2022078524A2 (en) 2021-11-03 2021-11-03 Specific conjugation of an antibody
AU2021362997A AU2021362997A1 (en) 2021-11-03 2021-11-03 Specific conjugation of an antibody
CN202280066606.8A CN118434765A (zh) 2021-11-03 2022-10-08 Bcma单克隆抗体和抗体-药物偶联物
EP22889047.1A EP4426741A4 (en) 2021-11-03 2022-10-08 Monoclonal Anti-BCMA Antibody and Antibody-Drug Conjugate
CA3236852A CA3236852A1 (en) 2021-11-03 2022-10-08 Bcma monoclonal antibody and the antibody-drug conjugate
JP2024526537A JP2024541059A (ja) 2021-11-03 2022-10-08 Ｂｃｍａモノクローナル抗体及び抗体薬物複合体
AU2022383265A AU2022383265A1 (en) 2021-11-03 2022-10-08 Bcma monoclonal antibody and the antibody-drug conjugate
PCT/CN2022/123901 WO2023078021A1 (en) 2021-11-03 2022-10-08 Bcma monoclonal antibody and the antibody-drug conjugate
KR1020247017883A KR20240095316A (ko) 2021-11-03 2022-10-08 Bcma 단일클론 항체 및 항체-약물 접합체
TW113142378A TW202506738A (zh) 2021-11-03 2022-10-28 抗體的特異性偶聯
TW111141166A TWI864472B (zh) 2021-11-03 2022-10-28 抗體的特異性偶聯
PCT/CN2022/129122 WO2023078273A1 (en) 2021-11-03 2022-11-02 Specific conjugation for an antibody-drug conjugate
CA3236754A CA3236754A1 (en) 2021-11-03 2022-11-02 Specific conjugation for an antibody-drug conjugate
EP22889295.6A EP4426729A1 (en) 2021-11-03 2022-11-02 Specific conjugation for an antibody-drug conjugate
AU2022381163A AU2022381163A1 (en) 2021-11-03 2022-11-02 Specific conjugation for an antibody-drug conjugate
CN202280069631.1A CN118215676A (zh) 2021-11-03 2022-11-02 抗体药物偶联物的特异性偶联
JP2024526538A JP2024542073A (ja) 2021-11-03 2022-11-02 抗体薬物結合体のための特異的共役
TW112138435A TWI923934B (zh) 2022-10-08 2023-10-06 Ｂｃｍａ 單株抗體及抗體藥物結合物
ZA2024/03371A ZA202403371B (en) 2021-11-03 2024-04-30 Specific conjugation of an antibody

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/CN2021/128453 WO2022078524A2 (en) 2021-11-03 2021-11-03 Specific conjugation of an antibody

Publications (3)

Publication Number Publication Date
WO2022078524A2 true WO2022078524A2 (en) 2022-04-21
WO2022078524A3 WO2022078524A3 (en) 2022-08-25
WO2022078524A4 WO2022078524A4 (en) 2022-12-08

Family

ID=81209474

Family Applications (3)

Application Number Title Priority Date Filing Date
PCT/CN2021/128453 Ceased WO2022078524A2 (en) 2021-11-03 2021-11-03 Specific conjugation of an antibody
PCT/CN2022/123901 Ceased WO2023078021A1 (en) 2021-11-03 2022-10-08 Bcma monoclonal antibody and the antibody-drug conjugate
PCT/CN2022/129122 Ceased WO2023078273A1 (en) 2021-11-03 2022-11-02 Specific conjugation for an antibody-drug conjugate

Family Applications After (2)

Application Number Title Priority Date Filing Date
PCT/CN2022/123901 Ceased WO2023078021A1 (en) 2021-11-03 2022-10-08 Bcma monoclonal antibody and the antibody-drug conjugate
PCT/CN2022/129122 Ceased WO2023078273A1 (en) 2021-11-03 2022-11-02 Specific conjugation for an antibody-drug conjugate

Country Status (9)

Country Link
EP (3) EP4426727A2 (https=)
JP (3) JP2024541058A (https=)
KR (2) KR20240095442A (https=)
CN (3) CN117980327A (https=)
AU (3) AU2021362997A1 (https=)
CA (3) CA3236930A1 (https=)
TW (2) TW202506738A (https=)
WO (3) WO2022078524A2 (https=)
ZA (1) ZA202403371B (https=)

Cited By (35)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115181076A (zh) * 2022-06-24 2022-10-14 北京丹大生物技术有限公司 一种用于检测阿立哌唑和脱氢阿立哌唑浓度的半抗原、抗原、细胞株、抗体、试剂和试剂盒
WO2023078273A1 (en) 2021-11-03 2023-05-11 Hangzhou Dac Biotech Co., Ltd. Specific conjugation for an antibody-drug conjugate
CN116773826A (zh) * 2023-08-21 2023-09-19 迪亚莱博（张家港）生物科技有限公司 一种用于检测抗蛋白酶3抗体的胶乳比浊生化试剂盒
CN116879425A (zh) * 2023-06-26 2023-10-13 盛禾(中国)生物制药有限公司 一种双特异性抗体蛋白纯度的检测方法
WO2023201268A1 (en) 2022-04-13 2023-10-19 Gilead Sciences, Inc. Combination therapy for treating tumor antigen expressing cancers
WO2023201267A1 (en) 2022-04-13 2023-10-19 Gilead Sciences, Inc. Combination therapy for treating trop-2 expressing cancers
WO2023222108A1 (zh) * 2022-05-20 2023-11-23 上海迈晋生物医药科技有限公司 抗体-药物偶联物的制备方法
WO2023230620A1 (en) * 2022-05-27 2023-11-30 Sonnet BioTherapeutics, Inc. Il-12-albumin-binding domain fusion protein formulations and methods of use thereof
CN117257977A (zh) * 2023-11-07 2023-12-22 正大天晴药业集团南京顺欣制药有限公司 用于制备抗体药物偶联物的方法
CN117304790A (zh) * 2023-11-27 2023-12-29 石狮佳南热熔胶有限公司 水性环保涂料及水性皮革
WO2024041545A1 (en) * 2022-08-22 2024-02-29 Suzhou Bioreinno Biotechnology Limited Company A novel thiol reductant, preparation method and use thereof
WO2024041544A1 (en) * 2022-08-22 2024-02-29 Suzhou Bioreinno Biotechnology Limited Company A method of preparing an antibody with site-specific modifications
WO2024041541A1 (en) * 2022-08-22 2024-02-29 Suzhou Bioreinno Biotechnology Limited Company A novel thiol reductant, method and use thereof
WO2024041543A1 (en) * 2022-08-22 2024-02-29 Suzhou Bioreinno Biotechnology Limited Company A method of preparing an antibody with thiol group site-specific modifications and use of tcep
WO2024020379A3 (en) * 2022-07-19 2024-03-14 Praesidia Biotherapeutics Inc. Prodrugs, prodrug compositions and related methods
WO2024054673A1 (en) * 2022-09-09 2024-03-14 Texas Tech University System Listeria monocytogenes as a vector for tumor-specific delivery of chemotherapeutic agents
WO2024051787A1 (zh) * 2022-09-09 2024-03-14 北京惠之衡生物科技有限公司 一种长效酰化胰岛素衍生物及其应用
JP2024519140A (ja) * 2021-05-27 2024-05-08 サイムワークス・ビーシー・インコーポレイテッド カンプトテシン類似体、複合体及び使用の方法
WO2024097812A1 (en) 2022-11-04 2024-05-10 Gilead Sciences, Inc. Therapy for treating bladder cancer
US12122826B2 (en) 2016-04-27 2024-10-22 Abbvie Inc. Methods of treatment of diseases in which IL-13 activity is detrimental using anti-IL-13 antibodies
WO2024225837A1 (ko) * 2023-04-28 2024-10-31 주식회사 에이비켐바이오 말레이미드 함유 링커-페이로드 접합체 및 이를 포함하는 신규한 항체-약물 중합체
US12134635B1 (en) 2023-12-29 2024-11-05 Sonnet BioTherapeutics, Inc. Interleukin 18 (IL-18) variants and fusion proteins comprising same
WO2024255873A1 (zh) * 2023-06-14 2024-12-19 江苏迈威康新药研发有限公司 一种制备抗体药物偶联物的方法
WO2025002310A1 (zh) * 2023-06-29 2025-01-02 石药集团巨石生物制药有限公司 抗体-药物偶联物及其用途
US12202857B2 (en) 2020-05-08 2025-01-21 Waters Technologies Corporation Methods for heat-assisted enzyme digestion
JP2025513310A (ja) * 2022-04-29 2025-04-24 シャンハイ ミクウルク ファーマシューティカル シーオー．エルティーディー カンプトテシン類似体のリガンド－薬物複合体、中間体、その製造方法、医薬組成物及びその用途
WO2025119322A1 (en) * 2023-12-07 2025-06-12 Wuxi Biologics (Shanghai) Co., Ltd. Gpc3 antibody-drug conjugates and use thereof
US12331130B2 (en) 2019-12-17 2025-06-17 The Board Of Regents Of The University Of Texas System DDR1 antibodies and uses thereof
WO2025151423A1 (en) * 2024-01-08 2025-07-17 OncoC4, Inc. Anti-cd24 antibody drug conjugates and uses thereof
JP2025524170A (ja) * 2022-07-29 2025-07-25 杭州愛科瑞思生物医薬有限公司 カンプトテシン誘導体及びその調製方法ならびに使用
US12411064B2 (en) 2020-05-08 2025-09-09 Waters Technologies Corporation Methods for heat-assisted enzyme digestion
WO2025207442A1 (en) * 2024-03-23 2025-10-02 Ranok Therapeutics (Hangzhou) Co. Ltd. Methods and compositions for targeted protein degradation
US12433952B2 (en) 2022-03-25 2025-10-07 Zymeworks Bc Inc. Antibody-drug conjugates targeting folate receptor alpha and methods of use
WO2025241603A1 (en) * 2023-05-24 2025-11-27 Hangzhou Seehe Biotechnology Co., Ltd An antibody drug conjugate with opened rings of thiosuccinimides, its preparation and application thereof
WO2026074296A1 (en) * 2024-10-03 2026-04-09 Iksuda Therapeutics Limited Linkers and protein and peptide conjugates incorporating the same

Families Citing this family (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
BR112023018676A2 (pt) 2021-03-18 2023-10-10 Seagen Inc Conjugado anticorpo-fármaco, composição farmacêutica, métodos de tratamento de uma doença ou condição e de um câncer, e, composição de conjugado ligante-fármaco
US11806405B1 (en) 2021-07-19 2023-11-07 Zeno Management, Inc. Immunoconjugates and methods
TW202320858A (zh) 2021-07-19 2023-06-01 美商薩諾管理公司 免疫接合物及方法
WO2023178289A2 (en) * 2022-03-17 2023-09-21 Seagen Inc. Camptothecin conjugates
EP4562051A1 (en) * 2022-07-26 2025-06-04 Immunome, Inc. Immunoconjugates and methods
CN115991750A (zh) * 2022-08-10 2023-04-21 北京大学人民医院 唾液素salivaricin在制备IL-21受体抑制剂中的应用
EP4735047A1 (en) * 2023-06-28 2026-05-06 Université de Tours Pegylated linkers and uses thereof
CN117069793A (zh) * 2023-08-08 2023-11-17 杭州爱科瑞思生物医药有限公司 双功能接头化合物、抗体药物偶联物及其制备方法和应用
KR20250043317A (ko) * 2023-09-21 2025-03-28 주식회사 피노바이오 운반체-약물 접합체 및 dnmt-1 저해제의 병용 요법
WO2025235686A1 (en) * 2024-05-07 2025-11-13 Aarvik Therapeutics, Inc. Fluorocamptothecin compounds, conjugates, and methods thereof
WO2025247261A1 (zh) * 2024-05-30 2025-12-04 爱尔利希生物科技有限公司 抗egfr抗体药物偶联物及其用途
CN118290514B (zh) * 2024-06-05 2025-04-04 东曜药业有限公司 基于阴离子膜的抗体偶联药物的纯化方法及抗体偶联药物
WO2026061476A1 (zh) * 2024-09-19 2026-03-26 长春金赛药业有限责任公司 一种多簇连接子、其制备方法及应用
CN120741749B (zh) * 2025-09-02 2025-11-21 凯莱英生命科学技术(天津)有限公司 抗体偶联药物中的抗体蛋白含量的检测方法
CN120837611B (zh) * 2025-09-22 2025-12-16 吉林大学 一种经鼻给药的司美格鲁肽制剂及其制备方法和应用

Family Cites Families (680)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US2292506A (en) 1940-03-29 1942-08-11 Gen Chemical Corp Manufacture of titanium salt solutions
FR901464A (fr) 1943-01-23 1945-07-27 Boehringer & Soehne Gmbh Procédé d'obtention de vanilline
CU22545A1 (es) 1994-11-18 1999-03-31 Centro Inmunologia Molecular Obtención de un anticuerpo quimérico y humanizado contra el receptor del factor de crecimiento epidérmico para uso diagnóstico y terapéutico
US4179337A (en) 1973-07-20 1979-12-18 Davis Frank F Non-immunogenic polypeptides
US4000304A (en) 1974-05-06 1976-12-28 Mead Johnson & Company Diuretic antiturombogenic and antiarrhythmic processes using N-substituted indole dimers and pyrrolobenzodia-zepine rearrangement products thereof
US4169888A (en) 1977-10-17 1979-10-02 The Upjohn Company Composition of matter and process
US4307016A (en) 1978-03-24 1981-12-22 Takeda Chemical Industries, Ltd. Demethyl maytansinoids
US4256746A (en) 1978-11-14 1981-03-17 Takeda Chemical Industries Dechloromaytansinoids, their pharmaceutical compositions and method of use
JPS55102583A (en) 1979-01-31 1980-08-05 Takeda Chem Ind Ltd 20-acyloxy-20-demethylmaytansinoid compound
JPS55162791A (en) 1979-06-05 1980-12-18 Takeda Chem Ind Ltd Antibiotic c-15003pnd and its preparation
JPS5645483A (en) 1979-09-19 1981-04-25 Takeda Chem Ind Ltd C-15003phm and its preparation
JPS5645485A (en) 1979-09-21 1981-04-25 Takeda Chem Ind Ltd Production of c-15003pnd
EP0028683A1 (en) 1979-09-21 1981-05-20 Takeda Chemical Industries, Ltd. Antibiotic C-15003 PHO and production thereof
US4491632A (en) 1979-10-22 1985-01-01 The Massachusetts General Hospital Process for producing antibodies to hepatitis virus and cell lines therefor
US4444887A (en) 1979-12-10 1984-04-24 Sloan-Kettering Institute Process for making human antibody producing B-lymphocytes
US4391904A (en) 1979-12-26 1983-07-05 Syva Company Test strip kits in immunoassays and compositions therein
EP0043718B1 (en) 1980-07-07 1984-11-28 National Research Development Corporation Improvements in or relating to cell lines
US4341761A (en) 1980-07-25 1982-07-27 E. I. Du Pont De Nemours And Company Antibodies to immunogenic peptides and their use to purify human fibroblast interferon
WO1982001188A1 (en) 1980-10-08 1982-04-15 Takeda Chemical Industries Ltd 4,5-deoxymaytansinoide compounds and process for preparing same
US4450254A (en) 1980-11-03 1984-05-22 Standard Oil Company Impact improvement of high nitrile resins
US4315929A (en) 1981-01-27 1982-02-16 The United States Of America As Represented By The Secretary Of Agriculture Method of controlling the European corn borer with trewiasine
US4313946A (en) 1981-01-27 1982-02-02 The United States Of America As Represented By The Secretary Of Agriculture Chemotherapeutically active maytansinoids from Trewia nudiflora
US4466917A (en) 1981-02-12 1984-08-21 New York University Malaria vaccine
US4493890A (en) 1981-03-23 1985-01-15 Miles Laboratories, Inc. Activated apoglucose oxidase and its use in specific binding assays
US4451570A (en) 1981-03-26 1984-05-29 The Regents Of The University Of California Immunoglobulin-secreting human hybridomas from a cultured human lymphoblastoid cell line
JPS57192389A (en) 1981-05-20 1982-11-26 Takeda Chem Ind Ltd Novel maytansinoid
US4414205A (en) 1981-08-28 1983-11-08 University Patents, Inc. Cell growth inhibitory substances
US4399121A (en) 1981-11-04 1983-08-16 Miles Laboratories, Inc. Iodothyronine immunogens and antibodies
US4427783A (en) 1981-12-14 1984-01-24 Hoffmann-La Roche Inc. Immunoassay of thymosin α1
US4671958A (en) 1982-03-09 1987-06-09 Cytogen Corporation Antibody conjugates for the delivery of compounds to target sites
US4464467A (en) 1982-07-26 1984-08-07 Bristol-Myers Company Antitumor antibiotics produced by new streptomyces
US4508647A (en) 1982-07-26 1985-04-02 Bristol-Myers Company Antitumor antibiotics BBM-2040A and BBM-2040B
US4427587A (en) 1982-11-10 1984-01-24 Bristol-Myers Company Total synthesis of antitumor antibiotics BBM-2040A and BBM-2040B
US4816567A (en) 1983-04-08 1989-03-28 Genentech, Inc. Recombinant immunoglobin preparations
US4663453A (en) 1983-05-18 1987-05-05 Hoechst-Roussel Pharmaceuticals Inc. Benzo[b]pyrrolo[3,2,1-jk][1,4]benzodiazepines having dopamine receptor activity
US4723003A (en) 1983-05-18 1988-02-02 Hoechst-Roussel Pharmaceuticals Inc. Benzopyrrolobenzodiazepines and quinobenzodiazepines
US4764616A (en) 1983-05-18 1988-08-16 Hoechst-Roussel Pharmaceuticals Inc. Benzopyrrolobenzodiazepines and quinobenzodiazepines
US4723007A (en) 1983-05-18 1988-02-02 Hoechst-Roussel Pharmaceuticals Inc. Benzopyrrolobenzodiazepines and quinobenzodiazepines
US4761412A (en) 1983-05-18 1988-08-02 Hoechst-Roussel Pharmaceuticals Inc. Benzopyrrolobenzodiazepines and quinobenzodiazepines useful for the treatment of psychoses
US4493795A (en) 1983-10-17 1985-01-15 Syntex (U.S.A.) Inc. Synthetic peptide sequences useful in biological and pharmaceutical applications and methods of manufacture
JPS6098584A (ja) 1983-11-02 1985-06-01 Canon Inc カメラ―体形ｖｔｒ
US6054561A (en) 1984-02-08 2000-04-25 Chiron Corporation Antigen-binding sites of antibody molecules specific for cancer antigens
US5169774A (en) 1984-02-08 1992-12-08 Cetus Oncology Corporation Monoclonal anti-human breast cancer antibodies
US4753894A (en) 1984-02-08 1988-06-28 Cetus Corporation Monoclonal anti-human breast cancer antibodies
US4912227A (en) 1984-02-21 1990-03-27 The Upjohn Company 1,2,8,8A-tetrahydrocyclopropa(c)pyrrolo(3,2-e)-indol-4-(5H)-ones and related compounds
US4978757A (en) 1984-02-21 1990-12-18 The Upjohn Company 1,2,8,8a-tetrahydrocyclopropa (C) pyrrolo [3,2-e)]-indol-4(5H)-ones and related compounds
US4764368A (en) 1984-08-29 1988-08-16 Dana-Farber Cancer Institute, Inc. Acid-cleavable compound
US4970198A (en) 1985-10-17 1990-11-13 American Cyanamid Company Antitumor antibiotics (LL-E33288 complex)
US4683230A (en) 1985-02-25 1987-07-28 Bristol-Myers Company BMY-28121, a new antitumor antibiotic
US4935362A (en) 1985-02-25 1990-06-19 Bristol-Myers Company BMY-28121, a new antitumor antibiotic
US5225539A (en) 1986-03-27 1993-07-06 Medical Research Council Recombinant altered antibodies and methods of making altered antibodies
JPH0684377B2 (ja) 1986-04-17 1994-10-26 協和醗酵工業株式会社 新規化合物ｄｃ―８８ａ及びｄｃ―８９ａ１
US6022958A (en) 1986-08-13 2000-02-08 Bayer Corporation cDNAs coding for members of the carcinoembryonic antigen family
US5332837A (en) 1986-12-19 1994-07-26 The Upjohn Company CC-1065 analogs
US5108912A (en) 1987-01-30 1992-04-28 American Cyanamid Company Antitumor antibiotics (LL-E33288 complex)
US4816444A (en) 1987-07-10 1989-03-28 Arizona Board Of Regents, Arizona State University Cell growth inhibitory substance
US4975278A (en) 1988-02-26 1990-12-04 Bristol-Myers Company Antibody-enzyme conjugates in combination with prodrugs for the delivery of cytotoxic agents to tumor cells
US5773435A (en) 1987-08-04 1998-06-30 Bristol-Myers Squibb Company Prodrugs for β-lactamase and uses thereof
US5770701A (en) 1987-10-30 1998-06-23 American Cyanamid Company Process for preparing targeted forms of methyltrithio antitumor agents
US5606040A (en) 1987-10-30 1997-02-25 American Cyanamid Company Antitumor and antibacterial substituted disulfide derivatives prepared from compounds possessing a methyl-trithio group
US5053394A (en) 1988-09-21 1991-10-01 American Cyanamid Company Targeted forms of methyltrithio antitumor agents
US4952394A (en) 1987-11-23 1990-08-28 Bristol-Myers Company Drug-monoclonal antibody conjugates
US4994578A (en) 1987-11-27 1991-02-19 Meiji Seika Kaisha, Ltd. Certain anti-tumor duocarmycin antibiotics from streptomyces
GB8800078D0 (en) 1988-01-05 1988-02-10 Ciba Geigy Ag Novel antibodies
IL106992A (en) 1988-02-11 1994-06-24 Bristol Myers Squibb Co Acylhydrazone derivatives of anthracycline and methods for their preparation
US5147786A (en) 1988-04-22 1992-09-15 Monsanto Company Immunoassay for the detection of α-haloacetamides
US5162504A (en) 1988-06-03 1992-11-10 Cytogen Corporation Monoclonal antibodies to a new antigenic marker in epithelial prostatic cells and serum of prostatic cancer patients
US4943628A (en) 1988-06-13 1990-07-24 Ortho Pharmaceutical Corporation HIV peptide-inducted T cell stimulation
JP2642165B2 (ja) 1988-07-22 1997-08-20 協和醗酵工業株式会社 新規ｄｃ−８９化合物およびその製造法
US5084468A (en) 1988-08-11 1992-01-28 Kyowa Hakko Kogyo Co., Ltd. Dc-88a derivatives
US5223409A (en) 1988-09-02 1993-06-29 Protein Engineering Corp. Directed evolution of novel binding proteins
AU4128089A (en) 1988-09-15 1990-03-22 Rorer International (Overseas) Inc. Monoclonal antibodies specific to human epidermal growth factor receptor and therapeutic methods employing same
GB8823869D0 (en) 1988-10-12 1988-11-16 Medical Res Council Production of antibodies
US5530101A (en) 1988-12-28 1996-06-25 Protein Design Labs, Inc. Humanized immunoglobulins
JP2598116B2 (ja) 1988-12-28 1997-04-09 協和醗酵工業株式会社 新規物質ｄｃ１１３
US4978744A (en) 1989-01-27 1990-12-18 Arizona Board Of Regents Synthesis of dolastatin 10
AU5186090A (en) 1989-02-23 1990-09-26 Colorado State University Research Foundation Gnrh analogs for destroying gonadotrophs
US4879278A (en) 1989-05-16 1989-11-07 Arizona Board Of Regents Isolation and structural elucidation of the cytostatic linear depsipeptide dolastatin 15
JP2510335B2 (ja) 1989-07-03 1996-06-26 協和醗酵工業株式会社 Ｄｃ―８８ａ誘導体
US5187186A (en) 1989-07-03 1993-02-16 Kyowa Hakko Kogyo Co., Ltd. Pyrroloindole derivatives
US5286637A (en) 1989-08-07 1994-02-15 Debiopharm, S.A. Biologically active drug polymer derivatives and method for preparing same
AU6355090A (en) 1989-08-23 1991-04-03 Scripps Clinic And Research Foundation Compositions and methods for detection and treatment of epstein-barr virus infection and immune disorders
US5495009A (en) 1989-10-24 1996-02-27 Gilead Sciences, Inc. Oligonucleotide analogs containing thioformacetal linkages
US5208020A (en) 1989-10-25 1993-05-04 Immunogen Inc. Cytotoxic agents comprising maytansinoids and their therapeutic use
US5334528A (en) 1989-10-30 1994-08-02 The Regents Of The University Of California Monoclonal antibodies to cyclodiene insecticides and method for detecting the same
US5165923A (en) 1989-11-20 1992-11-24 Imperial Cancer Research Technology Methods and compositions for the treatment of hodgkin's disease
US5859205A (en) 1989-12-21 1999-01-12 Celltech Limited Humanised antibodies
US6075181A (en) 1990-01-12 2000-06-13 Abgenix, Inc. Human antibodies derived from immunized xenomice
DE69120146T2 (de) 1990-01-12 1996-12-12 Cell Genesys Inc Erzeugung xenogener antikörper
US6673986B1 (en) 1990-01-12 2004-01-06 Abgenix, Inc. Generation of xenogeneic antibodies
US6150584A (en) 1990-01-12 2000-11-21 Abgenix, Inc. Human antibodies derived from immunized xenomice
HUT60768A (en) 1990-03-16 1992-10-28 Sandoz Ag Process for producing cd25 fixing molecules
CA2078118C (en) 1990-04-25 1999-11-16 Paul A. Aristoff Cc-1065 analogs
US5427908A (en) 1990-05-01 1995-06-27 Affymax Technologies N.V. Recombinant library screening methods
AU652936B2 (en) 1990-05-07 1994-09-15 Scripps Clinic And Research Foundation Intermediates in the formation of the calicheamicin and esperamicin oligosaccharides
US5767236A (en) 1990-05-09 1998-06-16 Biomeasure, Inc. Linear therapeutic peptides
US5137877B1 (en) 1990-05-14 1996-01-30 Bristol Myers Squibb Co Bifunctional linking compounds conjugates and methods for their production
US5256643A (en) 1990-05-29 1993-10-26 The Government Of The United States Human cripto protein
GB9014932D0 (en) 1990-07-05 1990-08-22 Celltech Ltd Recombinant dna product and method
US6172197B1 (en) 1991-07-10 2001-01-09 Medical Research Council Methods for producing members of specific binding pairs
GB9015198D0 (en) 1990-07-10 1990-08-29 Brien Caroline J O Binding substance
EP0468400B1 (en) 1990-07-26 1996-08-14 Kyowa Hakko Kogyo Co., Ltd. DC-89 derivatives as antitumoral agents
GB9017024D0 (en) 1990-08-03 1990-09-19 Erba Carlo Spa New linker for bioactive agents
US5633425A (en) 1990-08-29 1997-05-27 Genpharm International, Inc. Transgenic non-human animals capable of producing heterologous antibodies
US5770429A (en) 1990-08-29 1998-06-23 Genpharm International, Inc. Transgenic non-human animals capable of producing heterologous antibodies
US5661016A (en) 1990-08-29 1997-08-26 Genpharm International Inc. Transgenic non-human animals capable of producing heterologous antibodies of various isotypes
US5545806A (en) 1990-08-29 1996-08-13 Genpharm International, Inc. Ransgenic non-human animals for producing heterologous antibodies
ATE158021T1 (de) 1990-08-29 1997-09-15 Genpharm Int Produktion und nützung nicht-menschliche transgentiere zur produktion heterologe antikörper
US5625126A (en) 1990-08-29 1997-04-29 Genpharm International, Inc. Transgenic non-human animals for producing heterologous antibodies
US5789650A (en) 1990-08-29 1998-08-04 Genpharm International, Inc. Transgenic non-human animals for producing heterologous antibodies
US5877397A (en) 1990-08-29 1999-03-02 Genpharm International Inc. Transgenic non-human animals capable of producing heterologous antibodies of various isotypes
US5874299A (en) 1990-08-29 1999-02-23 Genpharm International, Inc. Transgenic non-human animals capable of producing heterologous antibodies
US5814318A (en) 1990-08-29 1998-09-29 Genpharm International Inc. Transgenic non-human animals for producing heterologous antibodies
AU9016591A (en) 1990-10-25 1992-05-26 Tanox Biosystems, Inc. Glycoproteins associated with membrane-bound immunoglobulins as antibody targets on B cells
CA2095633C (en) 1990-12-03 2003-02-04 Lisa J. Garrard Enrichment method for variant proteins with altered binding properties
ATE194385T1 (de) 1991-03-29 2000-07-15 Genentech Inc Menschliche pf4a rezeptoren und ihre verwendung
US5440021A (en) 1991-03-29 1995-08-08 Chuntharapai; Anan Antibodies to human IL-8 type B receptor
US5543503A (en) 1991-03-29 1996-08-06 Genentech Inc. Antibodies to human IL-8 type A receptor
US6797492B2 (en) 1991-05-17 2004-09-28 Merck & Co., Inc. Method for reducing the immunogenicity of antibody variable domains
EP1400536A1 (en) 1991-06-14 2004-03-24 Genentech Inc. Method for making humanized antibodies
WO1994004679A1 (en) 1991-06-14 1994-03-03 Genentech, Inc. Method for making humanized antibodies
JP3050424B2 (ja) 1991-07-12 2000-06-12 塩野義製薬株式会社 ヒトエンドセリンリセプター
US5264586A (en) 1991-07-17 1993-11-23 The Scripps Research Institute Analogs of calicheamicin gamma1I, method of making and using the same
WO1993003054A1 (fr) 1991-08-09 1993-02-18 Teikoku Hormone Mfg. Co., Ltd. Nouveau derive de tetrapeptide
US5264557A (en) 1991-08-23 1993-11-23 The United States Of America As Represented By The Department Of Health And Human Services Polypeptide of a human cripto-related gene, CR-3
US6153408A (en) 1991-11-15 2000-11-28 Institut Pasteur And Institut National De La Sante Et De La Recherche Medicale Altered major histocompatibility complex (MHC) determinant and methods of using the determinant
US6011146A (en) 1991-11-15 2000-01-04 Institut Pasteur Altered major histocompatibility complex (MHC) determinant and methods of using the determinant
PT1024191E (pt) 1991-12-02 2008-12-22 Medical Res Council Produção de auto-anticorpos a partir de reportórios de segmentos de anticorpo e exibidos em fagos
US5622929A (en) 1992-01-23 1997-04-22 Bristol-Myers Squibb Company Thioether conjugates
US5955075A (en) 1992-03-11 1999-09-21 Institute Of Virology, Slovak Academy Of Sciences Method of inhibiting tumor growth using antibodies to MN protein
EP0563475B1 (en) 1992-03-25 2000-05-31 Immunogen Inc Cell binding agent conjugates of derivatives of CC-1065
GB9314960D0 (en) 1992-07-23 1993-09-01 Zeneca Ltd Chemical compounds
JP3514490B2 (ja) 1992-08-21 2004-03-31 杏林製薬株式会社 トリフルオロメチルピロロインドールカルボン酸エステル誘導体及びその製造方法
US5288514A (en) 1992-09-14 1994-02-22 The Regents Of The University Of California Solid phase and combinatorial synthesis of benzodiazepine compounds on a solid support
US6015562A (en) 1992-09-22 2000-01-18 American Cyanamid Company Targeted forms of methyltrithio antitumor agents
US5324483B1 (en) 1992-10-08 1996-09-24 Warner Lambert Co Apparatus for multiple simultaneous synthesis
IL107366A (en) 1992-10-23 2003-03-12 Chugai Pharmaceutical Co Ltd Genes coding for megakaryocyte potentiator
US5736137A (en) 1992-11-13 1998-04-07 Idec Pharmaceuticals Corporation Therapeutic application of chimeric and radiolabeled antibodies to human B lymphocyte restricted differentiation antigen for treatment of B cell lymphoma
US5635483A (en) 1992-12-03 1997-06-03 Arizona Board Of Regents Acting On Behalf Of Arizona State University Tumor inhibiting tetrapeptide bearing modified phenethyl amides
US6034065A (en) 1992-12-03 2000-03-07 Arizona Board Of Regents Elucidation and synthesis of antineoplastic tetrapeptide phenethylamides of dolastatin 10
US6569834B1 (en) 1992-12-03 2003-05-27 George R. Pettit Elucidation and synthesis of antineoplastic tetrapeptide w-aminoalkyl-amides
US5644033A (en) 1992-12-22 1997-07-01 Health Research, Inc. Monoclonal antibodies that define a unique antigen of human B cell antigen receptor complex and methods of using same for diagnosis and treatment
US5410024A (en) 1993-01-21 1995-04-25 Arizona Board Of Regents Acting On Behalf Of Arizona State University Human cancer inhibitory pentapeptide amides
US5780588A (en) 1993-01-26 1998-07-14 Arizona Board Of Regents Elucidation and synthesis of selected pentapeptides
US5472693A (en) 1993-02-16 1995-12-05 The Dow Chemical Company Family of anti-carcinoembryonic antigen chimeric antibodies
US5436149A (en) 1993-02-19 1995-07-25 Barnes; Wayne M. Thermostable DNA polymerase with enhanced thermostability and enhanced length and efficiency of primer extension
US5801005A (en) 1993-03-17 1998-09-01 University Of Washington Immune reactivity to HER-2/neu protein for diagnosis of malignancies in which the HER-2/neu oncogene is associated
US5869445A (en) 1993-03-17 1999-02-09 University Of Washington Methods for eliciting or enhancing reactivity to HER-2/neu protein
US5475011A (en) 1993-03-26 1995-12-12 The Research Foundation Of State University Of New York Anti-tumor compounds, pharmaceutical compositions, methods for preparation thereof and for treatment
DE4314091A1 (de) 1993-04-29 1994-11-03 Boehringer Mannheim Gmbh Immunologisches Nachweisverfahren für Triazine
US6214345B1 (en) 1993-05-14 2001-04-10 Bristol-Myers Squibb Co. Lysosomal enzyme-cleavable antitumor drug conjugates
CA2163232A1 (en) 1993-05-20 1994-12-08 Robert C. Brasch Macromolecular contrast media for mr imaging
US5621039A (en) 1993-06-08 1997-04-15 Hallahan; Terrence W. Factor IX- polymeric conjugates
AU7113594A (en) 1993-06-21 1995-01-17 Enzon, Inc. Site specific synthesis of conjugated peptides
US5773223A (en) 1993-09-02 1998-06-30 Chiron Corporation Endothelin B1, (ETB1) receptor polypeptide and its encoding nucleic acid methods, and uses thereof
EP0731106B1 (en) 1993-10-01 2004-11-17 Teikoku Hormone Mfg. Co., Ltd. Dolastatin derivatives
GB9320575D0 (en) 1993-10-06 1993-11-24 Amp Gmbh Coaxial connector having improved locking mechanism
US5824701A (en) 1993-10-20 1998-10-20 Enzon, Inc. Taxane-based prodrugs
US5840900A (en) 1993-10-20 1998-11-24 Enzon, Inc. High molecular weight polymer-based prodrugs
AU8052194A (en) 1993-10-20 1995-05-08 Enzon, Inc. 2'- and/or 7- substituted taxoids
US5965566A (en) 1993-10-20 1999-10-12 Enzon, Inc. High molecular weight polymer-based prodrugs
US5880131A (en) 1993-10-20 1999-03-09 Enzon, Inc. High molecular weight polymer-based prodrugs
US5919455A (en) 1993-10-27 1999-07-06 Enzon, Inc. Non-antigenic branched polymer conjugates
US5643575A (en) 1993-10-27 1997-07-01 Enzon, Inc. Non-antigenic branched polymer conjugates
US5922545A (en) 1993-10-29 1999-07-13 Affymax Technologies N.V. In vitro peptide and antibody display libraries
US5951974A (en) 1993-11-10 1999-09-14 Enzon, Inc. Interferon polymer conjugates
US5786377A (en) 1993-11-19 1998-07-28 Universidad De Santiago De Compostela Pyrrolo 3,2-E!indol derivatives, process for the preparation thereof and applications
US5641780A (en) 1994-04-22 1997-06-24 Kyowa Hakko Kogyo Co., Ltd. Pyrrolo-indole derivatives
JPH07309761A (ja) 1994-05-20 1995-11-28 Kyowa Hakko Kogyo Co Ltd デュオカルマイシン誘導体の安定化法
US5773001A (en) 1994-06-03 1998-06-30 American Cyanamid Company Conjugates of methyltrithio antitumor agents and intermediates for their synthesis
US5730990A (en) 1994-06-24 1998-03-24 Enzon, Inc. Non-antigenic amine derived polymers and polymer conjugates
US5530097A (en) 1994-08-01 1996-06-25 Arizona Board Of Regents Acting On Behalf Of Arizona State University Human cancer inhibitory peptide amides
US5521284A (en) 1994-08-01 1996-05-28 Arizona Board Of Regents Acting On Behalf Of Arizona State University Human cancer inhibitory pentapeptide amides and esters
WO1996005863A1 (fr) 1994-08-19 1996-02-29 La Region Wallonne Composes, composition pharmaceutique et dispositif de diagnostic les comprenant et leur utilisation
US5554725A (en) 1994-09-14 1996-09-10 Arizona Board Of Regents Acting On Behalf Of Arizona State University Synthesis of dolastatin 15
US6111166A (en) 1994-09-19 2000-08-29 Medarex, Incorporated Transgenic mice expressing human Fcα and β receptors
US5750370A (en) 1995-06-06 1998-05-12 Human Genome Sciences, Inc. Nucleic acid encoding human endothlein-bombesin receptor and method of producing the receptor
US5738846A (en) 1994-11-10 1998-04-14 Enzon, Inc. Interferon polymer conjugates and process for preparing the same
US5599902A (en) 1994-11-10 1997-02-04 Arizona Board Of Regents Acting On Behalf Of Arizona State University Cancer inhibitory peptides
CN1051552C (zh) 1994-11-29 2000-04-19 杏林制药株式会社 丙烯酰胺衍生物及其制造方法
US5663149A (en) 1994-12-13 1997-09-02 Arizona Board Of Regents Acting On Behalf Of Arizona State University Human cancer inhibitory pentapeptide heterocyclic and halophenyl amides
ATE186745T1 (de) 1995-01-18 1999-12-15 Roche Diagnostics Gmbh Antikörper gegen cd30, die proteolytische spaltung und abgabe des membrangebundenen cd30 antigens verhindern
CN1113066C (zh) 1995-04-21 2003-07-02 帝国脏器制药株式会社 新型肽衍生物
EP0867190B1 (en) 1995-05-10 2007-12-26 Kyowa Hakko Kogyo Co., Ltd. Cytotoxin complexes comprising dipeptides
US5686292A (en) 1995-06-02 1997-11-11 Genentech, Inc. Hepatocyte growth factor receptor antagonist antibodies and uses thereof
US5686237A (en) 1995-06-05 1997-11-11 Al-Bayati; Mohammed A. S. Use of biomarkers in saliva to evaluate the toxicity of agents and the function of tissues in both biomedical and environmental applications
US6824993B2 (en) 1995-06-06 2004-11-30 Human Genome Sciences, Inc. Antibodies that bind human prostate specific G-protein receptor HPRAJ70
US5712374A (en) 1995-06-07 1998-01-27 American Cyanamid Company Method for the preparation of substantiallly monomeric calicheamicin derivative/carrier conjugates
US5714586A (en) 1995-06-07 1998-02-03 American Cyanamid Company Methods for the preparation of monomeric calicheamicin derivative/carrier conjugates
US5707829A (en) 1995-08-11 1998-01-13 Genetics Institute, Inc. DNA sequences and secreted proteins encoded thereby
US20020193567A1 (en) 1995-08-11 2002-12-19 Genetics Institute, Inc. Secreted proteins and polynucleotides encoding them
GB9517001D0 (en) 1995-08-18 1995-10-18 Denny William Enediyne compounds
SE9503380D0 (sv) 1995-09-29 1995-09-29 Pharmacia Ab Protein derivatives
AU727608B2 (en) 1995-10-03 2000-12-14 Scripps Research Institute, The CBI analogs of CC-1065 and the duocarmycins
EP0774464B1 (en) 1995-10-17 2004-07-28 Combichem, Inc. A template for solution phase synthesis of combinatorial libraries
DK0871490T3 (da) 1995-12-22 2003-07-07 Bristol Myers Squibb Co Forgrenede hydrazonlinkere
FR2742751B1 (fr) 1995-12-22 1998-01-30 Rhone Poulenc Rorer Sa Nouveaux taxoides, leur preparation et les compositions pharmaceutiques qui les contiennent
US6143901A (en) 1996-07-31 2000-11-07 Genesoft, Inc. Complex formation between dsDNA and pyrrole imidazole polyamides
EP0888301B1 (en) 1996-03-08 2005-08-10 The Scripps Research Institute Mcbi analogs of cc-1065 and the duocarmycins
JP3646191B2 (ja) 1996-03-19 2005-05-11 大塚製薬株式会社 ヒト遺伝子
WO1997034636A1 (en) 1996-03-20 1997-09-25 Immunomedics, Inc. Humanization of an anti-carcinoembryonic antigen anti-idiotype antibody and use as a tumor vaccine and for targeting applications
US6150508A (en) 1996-03-25 2000-11-21 Northwest Biotherapeutics, Inc. Monoclonal antibodies specific for the extracellular domain of prostate-specific membrane antigen
US7011812B1 (en) 1996-05-03 2006-03-14 Immunomedics, Inc. Targeted combination immunotherapy of cancer and infectious diseases
AU719513B2 (en) 1996-05-04 2000-05-11 Astrazeneca Ab Monoclonal antibody to CEA, conjugates comprising said antibody, and their therapeutic use in an adept system
US6107090A (en) 1996-05-06 2000-08-22 Cornell Research Foundation, Inc. Treatment and diagnosis of prostate cancer with antibodies to extracellur PSMA domains
EP0910636A1 (en) 1996-05-17 1999-04-28 Schering Corporation Human b-cell antigens, related reagents
EP0934269A4 (en) 1996-05-31 2000-01-12 Scripps Research Inst ANALOGS OF CC-1065 AND DUOCARMYCINE
US5728560A (en) 1996-06-14 1998-03-17 Enzon, Inc. Method of treating CD4+ T cell lymphopenia in immuno-compromised patients
US6590088B1 (en) 1996-07-19 2003-07-08 Human Genome Sciences, Inc. CD33-like protein
US5741892A (en) 1996-07-30 1998-04-21 Basf Aktiengesellschaft Pentapeptides as antitumor agents
US6130237A (en) 1996-09-12 2000-10-10 Cancer Research Campaign Technology Limited Condensed N-aclyindoles as antitumor agents
JPH1087666A (ja) 1996-09-18 1998-04-07 Kyorin Pharmaceut Co Ltd デュオカルマイシンｓａ及びその誘導体の製造中間体と製造方法
DE19638870B4 (de) 1996-09-23 2009-05-14 Helmholtz-Zentrum für Infektionsforschung GmbH Tubulysine, Verfahren zu ihrer Gewinnung und sie enthaltende Mittel
US5916771A (en) 1996-10-11 1999-06-29 Abgenix, Inc. Production of a multimeric protein by cell fusion method
US6653104B2 (en) 1996-10-17 2003-11-25 Immunomedics, Inc. Immunotoxins, comprising an internalizing antibody, directed against malignant and normal cells
US6417337B1 (en) 1996-10-31 2002-07-09 The Dow Chemical Company High affinity humanized anti-CEA monoclonal antibodies
US5880122A (en) 1996-11-01 1999-03-09 American Home Products Corporation 3-Carboxamide derivatives of 5H-pyrrolo 2,1-c! 1,4!-benzodiazepines
US6759509B1 (en) 1996-11-05 2004-07-06 Bristol-Myers Squibb Company Branched peptide linkers
US5989604A (en) 1996-12-19 1999-11-23 Adore-A-Pet, Ltd. Xylitol-containing non-human foodstuff and method
US5811452A (en) 1997-01-08 1998-09-22 The Research Foundation Of State University Of New York Taxoid reversal agents for drug-resistance in cancer chemotherapy and pharmaceutical compositions thereof
US5804183A (en) 1997-01-31 1998-09-08 Enzon, Inc. Arginine deminase derived from mycoplasma arthritidis and polymer conjugates containing the same
US5942602A (en) 1997-02-13 1999-08-24 Schering Aktiengessellschaft Growth factor receptor antibodies
US5945511A (en) 1997-02-20 1999-08-31 Zymogenetics, Inc. Class II cytokine receptor
US20030185830A1 (en) 1997-02-25 2003-10-02 Corixa Corporation Compositions and methods for the therapy and diagnosis of prostate cancer
WO1998036765A1 (en) 1997-02-25 1998-08-27 Arizona Board Of Regents Isolation and structural elucidation of the cytostatic linear and cyclo-depsipeptides dolastatin 16, dolastatin 17, and dolastatin 18
US7033827B2 (en) 1997-02-25 2006-04-25 Corixa Corporation Prostate-specific polynucleotide compositions
US6261791B1 (en) 1997-03-10 2001-07-17 The Regents Of The University Of California Method for diagnosing cancer using specific PSCA antibodies
US5965537A (en) 1997-03-10 1999-10-12 Basf Aktiengesellschaft Dolastatin 15 derivatives with carbonyl and heterocyclic functionalities at the C-terminus
EP0981541B1 (en) 1997-03-10 2004-06-02 The Regents Of The University Of California Psca: prostate stem cell antigen
US6541212B2 (en) 1997-03-10 2003-04-01 The Regents Of The University Of California Methods for detecting prostate stem cell antigen protein
US6306393B1 (en) 1997-03-24 2001-10-23 Immunomedics, Inc. Immunotherapy of B-cell malignancies using anti-CD22 antibodies
CA2285259A1 (en) 1997-03-28 1998-10-08 The Scripps Research Institute Sandramycin analogs
US6555339B1 (en) 1997-04-14 2003-04-29 Arena Pharmaceuticals, Inc. Non-endogenous, constitutively activated human protein-coupled receptors
US6319688B1 (en) 1997-04-28 2001-11-20 Smithkline Beecham Corporation Polynucleotide encoding human sodium dependent phosphate transporter (IPT-1)
US6235883B1 (en) 1997-05-05 2001-05-22 Abgenix, Inc. Human monoclonal antibodies to epidermal growth factor receptor
JP2002510968A (ja) 1997-05-07 2002-04-09 ブリストル−マイヤーズ スクイブ カンパニー 組換え抗体−酵素融合タンパク質
WO1998051805A1 (en) 1997-05-15 1998-11-19 Abbott Laboratories Reagents and methods useful for detecting diseases of the prostate
WO1998051824A1 (en) 1997-05-15 1998-11-19 Abbott Laboratories Reagents and methods useful for detecting disease of the urinary tract
NZ500789A (en) 1997-05-22 2002-05-31 Scripps Research Inst Analogs of duocarmycin and CC-1065
US5900402A (en) 1997-05-29 1999-05-04 Enzon, Inc. Method of reducing side effects associated with administration of oxygen-carrying proteins
US6143721A (en) 1997-07-18 2000-11-07 Basf Aktiengesellschaft Dolastatin 15 derivatives
US6011042A (en) 1997-10-10 2000-01-04 Enzon, Inc. Acyl polymeric derivatives of aromatic hydroxyl-containing compounds
WO1999019298A1 (en) 1997-10-14 1999-04-22 The Scripps Research Institute iso-CBI AND iso-CI ANALOGS OF CC-1065 AND THE DUOCARMYCINS
US20030060612A1 (en) 1997-10-28 2003-03-27 Genentech, Inc. Compositions and methods for the diagnosis and treatment of tumor
JPH11137291A (ja) 1997-11-12 1999-05-25 Mori Sangyo Kk アマニチン類の製造法
US20020034749A1 (en) 1997-11-18 2002-03-21 Billing-Medel Patricia A. Reagents and methods useful for detecting diseases of the breast
US6110695A (en) 1997-12-02 2000-08-29 The Regents Of The University Of California Modulating the interaction of the chemokine, B Lymphocyte Hemoattractant, and its Receptor, BLR1
NZ504884A (en) 1997-12-08 2002-10-25 Scripps Research Inst Synthesis of the dihydroindole C-ring of CC-1065/duocarmycin analogs and certain intermediates
US6180095B1 (en) 1997-12-17 2001-01-30 Enzon, Inc. Polymeric prodrugs of amino- and hydroxyl-containing bioactive agents
US5981709A (en) 1997-12-19 1999-11-09 Enzon, Inc. α-interferon-polymer-conjugates having enhanced biological activity and methods of preparing the same
US5965119A (en) 1997-12-30 1999-10-12 Enzon, Inc. Trialkyl-lock-facilitated polymeric prodrugs of amino-containing bioactive agents
WO1999035164A1 (en) 1998-01-09 1999-07-15 Arizona Board Of Regents, A Body Corporate, Acting On Behalf Of Arizona State University Anti-cryptococcal peptides
US6162930A (en) 1998-03-06 2000-12-19 Baylor University Anti-mitotic agents which inhibit tubulin polymerization
ES2313779T3 (es) 1998-03-13 2009-03-01 The Burnham Institute Moleculas que migran a diversos organos o tejidos seleccionados.
GB9809951D0 (en) 1998-05-08 1998-07-08 Univ Cambridge Tech Binding molecules
US6534482B1 (en) 1998-05-13 2003-03-18 Epimmune, Inc. Expression vectors for stimulating an immune response and methods of using the same
ATE473242T1 (de) 1998-05-20 2010-07-15 Immunomedics Inc Therapeutische mittel welche einen bispezifischen anti-hla class ii invariant chain x anti-pathogen antikörper enthalten
US20020187472A1 (en) 2001-03-09 2002-12-12 Preeti Lal Steap-related protein
US20030064397A1 (en) 1998-05-22 2003-04-03 Incyte Genomics, Inc. Transmembrane protein differentially expressed in prostate and lung tumors
US5981564A (en) 1998-07-01 1999-11-09 Universite Laval Water-soluble derivatives of paclitaxel, method for producing same and uses thereof
GB9818730D0 (en) 1998-08-27 1998-10-21 Univ Portsmouth Collections of compounds
GB9818732D0 (en) 1998-08-27 1998-10-21 Univ Portsmouth Collection of compounds
WO2000012130A1 (en) 1998-08-27 2000-03-09 Smithkline Beecham Corporation Rp105 agonists and antagonists
DE69925133T2 (de) 1998-08-27 2006-01-19 Spirogen Ltd., Ryde Pyrrolobenzodiazepine
GB9818731D0 (en) 1998-08-27 1998-10-21 Univ Portsmouth Compounds
JP4689781B2 (ja) 1998-09-03 2011-05-25 独立行政法人科学技術振興機構 アミノ酸輸送蛋白及びその遺伝子
JP3045706B1 (ja) 1998-09-14 2000-05-29 科学技術振興事業団 Ｄｎａの特定塩基配列をアルキル化する化合物及びその合成法
AU5963699A (en) 1998-10-02 2000-04-26 Mcmaster University Spliced form of (erb)b-2/neu oncogene
US6858710B2 (en) 1998-12-17 2005-02-22 Corixa Corporation Compositions and methods for the therapy and diagnosis of ovarian cancer
US20020119158A1 (en) 1998-12-17 2002-08-29 Corixa Corporation Compositions and methods for the therapy and diagnosis of ovarian cancer
US6468546B1 (en) 1998-12-17 2002-10-22 Corixa Corporation Compositions and methods for therapy and diagnosis of ovarian cancer
US20030091580A1 (en) 2001-06-18 2003-05-15 Mitcham Jennifer L. Compositions and methods for the therapy and diagnosis of ovarian cancer
US6962980B2 (en) 1999-09-24 2005-11-08 Corixa Corporation Compositions and methods for the therapy and diagnosis of ovarian cancer
DE69939553D1 (de) 1998-12-30 2008-10-23 Beth Israel Hospital Charakterisierung der proteinfamilie von soc/crac kalziumkanälen
US20030009013A1 (en) 1998-12-30 2003-01-09 Genentech, Inc. Secreted and transmembrane polypeptides and nucleic acids encoding the same
CZ20012587A3 (cs) 1999-01-29 2002-05-15 Corixa Corporation Izolovaný protein, nukleová kyselina, virový vektor, farmaceutický prostředek, izolovaná populace T buněk, způsob posílení imunitní odpovědi, způsob odstranění nádorových buněk, způsob stimulace a/nebo namnoľení T buněk a způsob přípravy fúzního proteinu
US6344451B1 (en) 1999-02-04 2002-02-05 American Home Products Pyrrolobenzodiazepine carboxyamide vasopressin agonists
GB9905124D0 (en) 1999-03-05 1999-04-28 Smithkline Beecham Biolog Novel compounds
AU3395900A (en) 1999-03-12 2000-10-04 Human Genome Sciences, Inc. Human lung cancer associated gene sequences and polypeptides
US6342219B1 (en) 1999-04-28 2002-01-29 Board Of Regents, The University Of Texas System Antibody compositions for selectively inhibiting VEGF
US7312318B2 (en) 2002-03-01 2007-12-25 Immunomedics, Inc. Internalizing anti-CD74 antibodies and methods of use
US7304126B2 (en) 1999-05-11 2007-12-04 Genentech, Inc. Secreted and transmembrane polypeptides and nucleic acids encoding the same
AU4952600A (en) 1999-06-03 2000-12-28 Takeda Chemical Industries Ltd. Screening method with the use of cd100
US7589172B2 (en) 1999-07-20 2009-09-15 Genentech, Inc. PRO256 polypeptides
NZ517331A (en) 1999-07-29 2004-02-27 Medarex Inc Human monoclonal antibodies to prostate specific membrane antigen and a transgenic mouse to produce the antibody
US7297770B2 (en) 1999-08-10 2007-11-20 Genentech, Inc. PRO6496 polypeptides
US7294696B2 (en) 1999-08-17 2007-11-13 Genentech Inc. PRO7168 polypeptides
US6716966B1 (en) 1999-08-18 2004-04-06 Altarex Corp. Therapeutic binding agents against MUC-1 antigen and methods for their use
US7147850B2 (en) 1999-08-18 2006-12-12 Altarex Medical Corp. Therapeutic binding agents against MUC-1 antigen and methods for their use
US6909006B1 (en) 1999-08-27 2005-06-21 Spirogen Limited Cyclopropylindole derivatives
CA2380355A1 (en) 1999-09-01 2001-03-08 Genentech, Inc. Secreted and transmembrane polypeptides and nucleic acids encoding the same
US20030206918A1 (en) 1999-09-10 2003-11-06 Corixa Corporation Compositions and methods for the therapy and diagnosis of ovarian cancer
US6323315B1 (en) 1999-09-10 2001-11-27 Basf Aktiengesellschaft Dolastatin peptides
US20030232056A1 (en) 1999-09-10 2003-12-18 Corixa Corporation Compositions and methods for the therapy and diagnosis of ovarian cancer
US20030129192A1 (en) 1999-09-10 2003-07-10 Corixa Corporation Compositions and methods for the therapy and diagnosis of ovarian cancer
EP1089473A1 (en) 1999-09-28 2001-04-04 TELEFONAKTIEBOLAGET L M ERICSSON (publ) Apparatus and method for time-aligning data frames of a plurality of channels in a telecommunication system
US7303749B1 (en) 1999-10-01 2007-12-04 Immunogen Inc. Compositions and methods for treating cancer using immunoconjugates and chemotherapeutic agents
ES2309012T3 (es) 1999-10-29 2008-12-16 Genentech, Inc. Composiciones del anticuerpo anti-psca y a sus procedimientos contra celulas cancerigenas que expresen psca.
CA2388063C (en) 1999-11-24 2010-06-08 Immunogen, Inc. Cytotoxic agents comprising taxanes and their therapeutic use
AU2048901A (en) 1999-11-29 2001-06-04 Trustees Of Columbia University In The City Of New York, The Isolation of five novel genes coding for new Fc receptors-type melanoma involved in the pathogenesis of lymphoma/melanoma
EP1248800A2 (en) 1999-11-30 2002-10-16 Corixa Corporation Compositions and methods for therapy and diagnosis of breast cancer
EP1239866A4 (en) 1999-12-10 2005-02-09 Epimmune Inc INDUCTION OF HER2 / NEU CELLULAR IMMUNE RESPONSES USING PEPTIDE AND NUCLEIC ACID-CONTAINING COMPOSITIONS
AU783473B2 (en) 1999-12-23 2005-10-27 Zymogenetics Inc. Soluble interleukin-20 receptor
NZ502058A (en) 1999-12-23 2003-11-28 Ovita Ltd Isolated mutated nucleic acid molecule for regulation of ovulation rate
EP1244708B8 (en) 1999-12-23 2007-03-07 ZymoGenetics, Inc. Method for treating inflammation
US6610286B2 (en) 1999-12-23 2003-08-26 Zymogenetics, Inc. Method for treating inflammation using soluble receptors to interleukin-20
AU767394C (en) 1999-12-29 2005-04-21 Immunogen, Inc. Cytotoxic agents comprising modified doxorubicins and daunorubicins and their therapeutic use
IT1317731B1 (it) 2000-01-18 2003-07-15 Indena Spa Tassani semisintetici con attivita' antitumorale ed antiangiogenetica.
US7294695B2 (en) 2000-01-20 2007-11-13 Genentech, Inc. PRO10268 polypeptides
CA2398102A1 (en) 2000-01-21 2001-07-26 Corixa Corporation Compounds and methods for prevention and treatment of her-2/neu associated malignancies
US20030224379A1 (en) 2000-01-21 2003-12-04 Tang Y. Tom Novel nucleic acids and polypeptides
AU2001243142A1 (en) 2000-02-03 2001-08-14 Hyseq, Inc. Novel nucleic acids and polypeptides
US20020142377A1 (en) 2000-02-22 2002-10-03 Glucksmann Maria Alexandra 18607, a novel human calcium channel
US20030219806A1 (en) 2000-02-22 2003-11-27 Millennium Pharmaceuticals, Inc. Novel 18607, 15603, 69318, 12303, 48000, 52920, 5433, 38554, 57301, 58324, 55063, 52991, 59914, 59921 and 33751 molecules and uses therefor
DE10008089A1 (de) 2000-02-22 2001-10-31 Biotechnolog Forschung Gmbh Syntheseverfahren zur Herstellung von Tubulysinen
ATE419354T1 (de) 2000-02-25 2009-01-15 Us Gov Health & Human Serv Scfv-moleküle gegen egfrviii mit verbesserter zytotoxizität und ausbeute, darauf basierte immuntoxine, und verfahren zur deren verwendung
US20040002068A1 (en) 2000-03-01 2004-01-01 Corixa Corporation Compositions and methods for the detection, diagnosis and therapy of hematological malignancies
US20040005561A1 (en) 2000-03-01 2004-01-08 Corixa Corporation Compositions and methods for the detection, diagnosis and therapy of hematological malignancies
EP1261743A2 (en) 2000-03-07 2002-12-04 Hyseq, Inc. Novel nucleic acids and polypeptides
US7097840B2 (en) 2000-03-16 2006-08-29 Genentech, Inc. Methods of treatment using anti-ErbB antibody-maytansinoid conjugates
CA2403537A1 (en) 2000-03-16 2001-10-11 Genesoft, Inc. Charged compounds comprising a nucleic acid binding moiety and uses therefor
AU2001249411B2 (en) 2000-03-24 2007-02-15 Fahri Saatcioglu Novel prostate-specific or testis-specific nucleic acid molecules, polypeptides,and diagnostic and therapeutic methods
WO2004043361A2 (en) 2002-11-08 2004-05-27 Genentech, Inc. Compositions and methods for the treatment of natural killer cell related diseases
AU2001250412A1 (en) 2000-03-31 2001-10-08 Ipf Pharmaceuticals Gmbh Diagnostic and medicament for analysing the cell surface proteome of tumour and inflammatory cells and for treating tumorous and inflammatory diseases, preferably using specific chemokine receptor analysis and the chemokine receptor-ligand interaction
US7279294B2 (en) 2000-04-03 2007-10-09 The United States Of America As Represented By The Secretary, Dept. Of Health And Human Services, Nih Tumor markers in ovarian cancer
CN1420929A (zh) 2000-04-07 2003-05-28 阿瑞那制药公司 非内源性组成型活化的已知g蛋白偶联受体
JP4061819B2 (ja) 2000-05-12 2008-03-19 独立行政法人科学技術振興機構 インターストランドクロスリンク剤の合成方法
US20030119115A1 (en) 2000-05-17 2003-06-26 Genentech, Inc. Secreted and transmembrane polypeptides and nucleic acids encoding the same
WO2001088133A2 (en) 2000-05-18 2001-11-22 Lexicon Genetics Incorporated Human semaphorin homologs and polynucleotides encoding the same
AU2001274888A1 (en) 2000-05-19 2001-12-03 Human Genome Sciences, Inc. Nucleic acids, proteins, and antibodies
AU2001275437A1 (en) 2000-06-09 2001-12-17 Idec Pharmaceuticals Corporation Gene targets and ligands that bind thereto for treatment and diagnosis of ovarian carcinomas
AU2001266853B2 (en) 2000-06-14 2005-02-17 Medarex, Inc. Prodrug compounds with an oligopeptide having an isoleucine residue
JP2004501875A (ja) 2000-06-14 2004-01-22 メダレックス，インコーポレイティド トリペプチドプロドラッグ化合物
WO2001098351A2 (en) 2000-06-16 2001-12-27 Incyte Genomics, Inc. G-protein coupled receptors
EP1383892A2 (en) 2000-06-30 2004-01-28 Incyte Genomics, Inc. Human extracellular matrix and cell adhesion polypeptides
WO2002002624A2 (en) 2000-06-30 2002-01-10 Amgen, Inc. B7-like molecules and uses thereof
EP1301524A1 (en) 2000-06-30 2003-04-16 Human Genome Sciences, Inc. B7-like polynucleotides, polypeptides, and antibodies
WO2002006339A2 (en) 2000-07-03 2002-01-24 Curagen Corporation Proteins and nucleic acids encoding same
US20040044179A1 (en) 2000-07-25 2004-03-04 Genentech, Inc. Secreted and transmembrane polypeptides and nucleic acids encoding the same
US6891030B2 (en) 2000-07-27 2005-05-10 Mayo Foundation For Medical Education And Research T-cell immunoregulatory molecule
DE60134178D1 (de) 2000-07-28 2008-07-03 Ulrich Wissenbach Trp8 krebsmarker
US7229623B1 (en) 2000-08-03 2007-06-12 Corixa Corporation Her-2/neu fusion proteins
US7163681B2 (en) 2000-08-07 2007-01-16 Centocor, Inc. Anti-integrin antibodies, compositions, methods and uses
CA2418014C (en) 2000-08-08 2014-01-21 Immunomedics, Inc. Immunotherapy for chronic myelocytic leukemia
EP1366153A2 (en) 2000-08-14 2003-12-03 Corixa Corporation Compositions and methods for the therapy and diagnosis of her-2/neu-associated malignancies
WO2002013847A2 (en) 2000-08-14 2002-02-21 Corixa Corporation Methods for diagnosis and therapy of hematological and virus-associated malignancies
US6333410B1 (en) 2000-08-18 2001-12-25 Immunogen, Inc. Process for the preparation and purification of thiol-containing maytansinoids
GB0020953D0 (en) 2000-08-24 2000-10-11 Smithkline Beecham Biolog Vaccine
CA2420140A1 (en) 2000-08-24 2002-02-28 Genentech, Inc. Compositions and methods for the diagnosis and treatment of tumor
EP1346040A2 (en) 2000-09-11 2003-09-24 Nuvelo, Inc. Novel nucleic acids and polypeptides
US6613567B1 (en) 2000-09-15 2003-09-02 Isis Pharmaceuticals, Inc. Antisense inhibition of Her-2 expression
ATE333888T1 (de) 2000-09-15 2006-08-15 Zymogenetics Inc Verwendung eines polypeptids, welches die extrazelluläre domäne von il-20ra und il-20rb enthält, zur behandlung von entzündungen
US7582293B2 (en) 2000-09-15 2009-09-01 Genentech, Inc. Anti-PRO6308 antibodies
NZ525380A (en) 2000-09-18 2008-06-30 Biogen Idec Inc Non-fucosylated forms of Cripto and their use as tumor blocking agents
UA83458C2 (uk) 2000-09-18 2008-07-25 Байоджен Айдек Ма Інк. Виділений поліпептид baff-r (рецептор фактора активації в-клітин сімейства tnf)
EP1474528A4 (en) 2000-10-13 2006-06-14 Protein Design Labs Inc METHODS OF DIAGNOSIS OF PROSTATE CANCER, COMPOSITIONS AND METHODS OF EXAMINING PROSTATE CANCER MODULATORS
US6596541B2 (en) 2000-10-31 2003-07-22 Regeneron Pharmaceuticals, Inc. Methods of modifying eukaryotic cells
ES2329012T3 (es) 2000-11-07 2009-11-20 Zymogenetics, Inc. Receptor del factor de necrosis tumoral humano.
US20020150573A1 (en) 2000-11-10 2002-10-17 The Rockefeller University Anti-Igalpha-Igbeta antibody for lymphoma therapy
US7090843B1 (en) 2000-11-28 2006-08-15 Seattle Genetics, Inc. Recombinant anti-CD30 antibodies and uses thereof
WO2002061087A2 (en) 2000-12-19 2002-08-08 Lifespan Biosciences, Inc. Antigenic peptides, such as for g protein-coupled receptors (gpcrs), antibodies thereto, and systems for identifying such antigenic peptides
WO2002054940A2 (en) 2001-01-12 2002-07-18 University Of Medicine & Dentistry Of New Jersey Bone morphogenetic protein-2 in the treatment and diagnosis of cancer
US20030119125A1 (en) 2001-01-16 2003-06-26 Genentech, Inc. Secreted and transmembrane polypeptides and nucleic acids encoding the same
US20030119126A1 (en) 2001-01-16 2003-06-26 Genentech, Inc. Secreted and transmembrane polypeptides and nucleic acids encoding the same
US7754208B2 (en) 2001-01-17 2010-07-13 Trubion Pharmaceuticals, Inc. Binding domain-immunoglobulin fusion proteins
JP2005503760A (ja) 2001-01-24 2005-02-10 プロテイン デザイン ラブス， インコーポレイテッド 乳癌の診断方法、組成物および乳癌のモジュレーターのスクリーニング方法
WO2002060317A2 (en) 2001-01-30 2002-08-08 Corixa Corporation Compositions and methods for the therapy and diagnosis of pancreatic cancer
WO2002064780A1 (en) 2001-02-12 2002-08-22 Bionomics Limited Dna sequences for human tumour suppressor genes
AU2002258518A1 (en) 2001-03-14 2002-09-24 Millennium Pharmaceuticals, Inc. Nucleic acid molecules and proteins for the identification, assessment, prevention, and therapy of ovarian cancer
US20020169125A1 (en) 2001-03-21 2002-11-14 Cell Therapeutics, Inc. Recombinant production of polyanionic polymers and uses thereof
EP1243276A1 (en) 2001-03-23 2002-09-25 Franciscus Marinus Hendrikus De Groot Elongated and multiple spacers containing activatible prodrugs
US20040236091A1 (en) 2001-03-28 2004-11-25 Chicz Roman M. Translational profiling
JP2005508839A (ja) 2001-03-29 2005-04-07 ラモット・アット・テル・アビブ・ユニバーシテイ・リミテッド ペプチドおよびｍｕｃ１タンパク質に対する抗体
JP2004536578A (ja) 2001-03-30 2004-12-09 ザ リージェンツ オブ ザ ユニバーシティ オブ カリフォルニア 腫瘍ターゲティング用の抗−ｍｕｃ−１単鎖抗体
US7183388B2 (en) 2001-03-30 2007-02-27 The Regents Of The University Of California Anti-MUC-1 single chain antibodies for tumor targeting
WO2003008537A2 (en) 2001-04-06 2003-01-30 Mannkind Corporation Epitope sequences
US6820011B2 (en) 2001-04-11 2004-11-16 The Regents Of The University Of Colorado Three-dimensional structure of complement receptor type 2 and uses thereof
US7202239B2 (en) 2001-04-12 2007-04-10 Wyeth Cyclohexylphenyl carboxamides tocolytic oxytocin receptor antagonists
US7064120B2 (en) 2001-04-12 2006-06-20 Wyeth Tricyclic pyridyl carboxamides and derivatives thereof tocolytic oxytocin receptor antagonists
US7326700B2 (en) 2001-04-12 2008-02-05 Wyeth Cyclohexenyl phenyl carboxamides tocolytic oxytocin receptor antagonists
US7022699B2 (en) 2001-04-12 2006-04-04 Wyeth Cyclohexenyl phenyl diazepines vasopressin and oxytocin receptor modulators
US7109193B2 (en) 2001-04-12 2006-09-19 Wyeth Tricyclic diazepines tocolytic oxytocin receptor antagonists
US6977254B2 (en) 2001-04-12 2005-12-20 Wyeth Hydroxy cyclohexenyl phenyl carboxamides tocolytic oxytocin receptor antagonists
CA2443617C (en) 2001-04-17 2013-12-10 The Board Of Trustees Of The University Of Arkansas Repeat sequences of the ca125 gene and their use for diagnostic and therapeutic interventions
AU2002309583A1 (en) 2001-04-18 2002-11-05 Protein Desing Labs, Inc. Methods of diagnosis of lung cancer, compositions and methods of screening for modulators of lung cancer
PL207087B1 (pl) 2001-04-26 2010-10-29 Biogen Przeciwciało, które wiąże się swoiście z białkiem Cripto, zawierająca je kompozycja farmaceutyczna i zastosowanie przeciwciała
US6884869B2 (en) 2001-04-30 2005-04-26 Seattle Genetics, Inc. Pentapeptide compounds and uses related thereto
JP2005504513A (ja) 2001-05-09 2005-02-17 コリクサ コーポレイション 前立腺癌の治療及び診断のための組成物及び方法
US20030078399A1 (en) 2001-05-11 2003-04-24 Sloan-Kettering Institute For Cancer Research Nucleic acid sequence encoding ovarian antigen, CA125, and uses thereof
BRPI0209933B8 (pt) 2001-05-24 2021-05-25 Zymogenetics Inc proteína de fusão, e, molécula de ácido nucleico
US6989452B2 (en) 2001-05-31 2006-01-24 Medarex, Inc. Disulfide prodrugs and linkers and stabilizers useful therefor
US6441163B1 (en) 2001-05-31 2002-08-27 Immunogen, Inc. Methods for preparation of cytotoxic conjugates of maytansinoids and cell binding agents
MXPA03010961A (es) 2001-05-31 2004-02-27 Vertex Pharma Compuestos de tiazol utiles como inhibidores de proteinas cinasas.
WO2002098897A2 (en) 2001-06-01 2002-12-12 Cornell Research Foundation, Inc. Modified antibodies to prostate-specific membrane antigen and uses thereof
US7157558B2 (en) 2001-06-01 2007-01-02 Genentech, Inc. Polypeptide encoded by a polynucleotide overexpresses in tumors
AU2002314901A1 (en) 2001-06-04 2002-12-16 Eos Biotechnology, Inc. Methods of diagnosis and treatment of androgen-dependent prostate cancer, prostate cancer undergoing androgen-withdrawal, and androgen-independent prostate cancer
JP2005518185A (ja) 2001-06-04 2005-06-23 キュラジェン コーポレイション 新規タンパク質およびそれをコード化する核酸
DE60237917D1 (de) 2001-06-05 2010-11-18 Exelixis Inc Gfats als modifikatoren des p53-wegs und verwendungsverfahren
EP1402053A4 (en) 2001-06-05 2005-05-11 Exelixis Inc CHDS AS MODULATORS OF THE P53 PATH AND METHOD OF USE
US7235358B2 (en) 2001-06-08 2007-06-26 Expression Diagnostics, Inc. Methods and compositions for diagnosing and monitoring transplant rejection
WO2002100353A2 (en) 2001-06-11 2002-12-19 Medarex, Inc. Cd10-activated prodrug compounds
WO2002101075A2 (en) 2001-06-13 2002-12-19 Millennium Pharmaceuticals, Inc. Novel genes, compositions, kits, and methods for identification, assessment, prevention, and therapy of cervical cancer
US7189507B2 (en) 2001-06-18 2007-03-13 Pdl Biopharma, Inc. Methods of diagnosis of ovarian cancer, compositions and methods of screening for modulators of ovarian cancer
WO2002102235A2 (en) 2001-06-18 2002-12-27 Eos Biotechnology Inc. Methods of diagnosis of ovarian cancer, compositions and methods of screening for modulators of ovarian cancer
WO2003004989A2 (en) 2001-06-21 2003-01-16 Millennium Pharmaceuticals, Inc. Compositions, kits, and methods for identification, assessment, prevention, and therapy of breast cancer
AU2002315411A1 (en) 2001-06-21 2003-01-08 Glycotech Corporation Detection and treatment of prostate cancer
US20030108958A1 (en) 2001-06-28 2003-06-12 Rene De Waal Malefyt Biological activity of AK155
WO2003004529A2 (en) 2001-07-02 2003-01-16 Licentia Ltd. Ephrin-tie receptor materials and methods
US20040076955A1 (en) 2001-07-03 2004-04-22 Eos Biotechnology, Inc. Methods of diagnosis of bladder cancer, compositions and methods of screening for modulators of bladder cancer
WO2003003984A2 (en) 2001-07-05 2003-01-16 Curagen Corporation Novel proteins and nucleic acids encoding same
WO2003055439A2 (en) 2001-07-18 2003-07-10 The Regents Of The University Of California Her2/neu target antigen and use of same to stimulate an immune response
WO2003009814A2 (en) 2001-07-25 2003-02-06 Millennium Pharmaceuticals, Inc. Novel genes, compositions, kits, and methods for identification, assessment, prevention, and therapy of prostate cancer
NZ571596A (en) 2001-08-03 2010-11-26 Glycart Biotechnology Ag Antibody glycosylation variants having increased antibody-dependent cellular cytotoxicity
BR0211614A (pt) 2001-08-03 2006-10-31 Genentech Inc polipeptìdeo tacis e br3 e empregos dos mesmos
AU2002324700A1 (en) 2001-08-14 2003-03-03 Bayer Ag Nucleic acid and amino acid sequences involved in pain
US20030092013A1 (en) 2001-08-16 2003-05-15 Vitivity, Inc. Diagnosis and treatment of vascular disease
WO2003018621A2 (en) 2001-08-23 2003-03-06 Oxford Biomedica (Uk) Limited Genes
US6902930B2 (en) 2001-08-29 2005-06-07 Vanderbilt University Human Mob-5 (IL-24) receptors and uses thereof
US20030124579A1 (en) 2001-09-05 2003-07-03 Eos Biotechnology, Inc. Methods of diagnosis of ovarian cancer, compositions and methods of screening for modulators of ovarian cancer
EP2287186B1 (en) 2001-09-06 2014-12-31 Agensys, Inc. Nucleic acid and corresponding protein entitled STEAP-1 useful in treatment and detection of cancer
US7098305B2 (en) 2001-09-06 2006-08-29 Ardana Bioscience Limited Sustained release of microcrystalline peptide suspensions
AU2002330039A1 (en) 2001-09-17 2003-04-01 Eos Biotechnology, Inc. Methods of diagnosis of cancer compositions and methods of screening for modulators of cancer
US20050004017A1 (en) 2001-09-18 2005-01-06 Yuval Reiss Methods and compositions for treating hcap associated diseases
DE60238143D1 (de) 2001-09-18 2010-12-09 Genentech Inc Zusammensetzungen und verfahren für die diagnose von tumoren
CA2460621A1 (en) 2001-09-19 2003-03-27 Nuvelo, Inc. Novel nucleic acids and polypeptides
US7091186B2 (en) 2001-09-24 2006-08-15 Seattle Genetics, Inc. p-Amidobenzylethers in drug delivery agents
US20030077644A1 (en) 2001-09-28 2003-04-24 Bing Yang Diagnosis and treatment of diseases caused by mutations in CD72
US20050130117A1 (en) 2001-10-03 2005-06-16 Davis Cong L. Modulators of lymphocyte activation and migration
AU2002362454A1 (en) 2001-10-03 2003-04-14 Origene Technologies, Inc. Regulated breast cancer genes
AR036833A1 (es) 2001-10-18 2004-10-06 Bayer Corp Anticuerpos humanos que se unen a mn y tienen actividad neutralizante de la adhesion celular.
US20050123925A1 (en) 2002-11-15 2005-06-09 Genentech, Inc. Compositions and methods for the diagnosis and treatment of tumor
WO2003034984A2 (en) 2001-10-19 2003-05-01 Genentech, Inc. Compositions and methods for the diagnosis and treatment of inflammatory bowel disorders
JP5355839B2 (ja) 2001-10-23 2013-11-27 ピーエスエムエー デベロプメント カンパニー， エル．エル．シー． Ｐｓｍａ抗体およびタンパク質マルチマー
CA2465268A1 (en) 2001-10-24 2003-05-01 National Jewish Medical And Research Center Three-dimensional structures of tall-1 and its cognate receptors and modified proteins and methods related thereto
KR100996226B1 (ko) 2001-10-31 2010-11-24 유니버시티 오브 노스 텍사스 헬스 사이언스 센터 뼈 형태발생 단백질(ｂｍｐ), ｂｍｐ 수용체 및 ｂｍｐ결합 단백질 및 녹내장 진단 및 치료에서 그의 용도
WO2003042661A2 (en) 2001-11-13 2003-05-22 Protein Design Labs, Inc. Methods of diagnosis of cancer, compositions and methods of screening for modulators of cancer
US20030232350A1 (en) 2001-11-13 2003-12-18 Eos Biotechnology, Inc. Methods of diagnosis of cancer, compositions and methods of screening for modulators of cancer
AU2002339691A1 (en) 2001-11-29 2003-06-10 Genset Agonists and antagonists of prolixin for the treatment of metabolic disorders
AU2002351208A1 (en) 2001-12-03 2003-06-17 Abgenix, Inc. Antibodies against carbonic anhydrase IX (CA IX) tumor antigen
AU2002349784A1 (en) 2001-12-03 2003-06-17 Asahi Kasei Pharma Corporation Nf-kappab activating genes
AU2002366951A1 (en) 2001-12-10 2003-07-09 Nuvelo,Inc. Novel nucleic acids and polypeptides
US6716821B2 (en) 2001-12-21 2004-04-06 Immunogen Inc. Cytotoxic agents bearing a reactive polyethylene glycol moiety, cytotoxic conjugates comprising polyethylene glycol linking groups, and methods of making and using the same
US20030134790A1 (en) 2002-01-11 2003-07-17 University Of Medicine And Dentistry Of New Jersey Bone Morphogenetic Protein-2 And Bone Morphogenetic Protein-4 In The Treatment And Diagnosis Of Cancer
US7452675B2 (en) 2002-01-25 2008-11-18 The Queen's Medical Center Methods of screening for TRPM4b modulators
CA2474616A1 (en) 2002-01-28 2003-08-07 Medarex, Inc. Human monoclonal antibodies to prostate specific membrane antigen (psma)
WO2003072736A2 (en) 2002-02-21 2003-09-04 Duke University Reagents and treatment methods for autoimmune diseases
HUE026218T2 (en) 2002-02-21 2016-05-30 Inst Virology MN / CA IX-Specific Monoclonal Antibodies and Methods for MN / CA IX Deficient Mice
EP1575480A4 (en) 2002-02-22 2008-08-06 Genentech Inc COMPOSITIONS AND METHODS OF TREATING IMMUNE-INFLAMMATORY DISEASES
US7591994B2 (en) 2002-12-13 2009-09-22 Immunomedics, Inc. Camptothecin-binding moiety conjugates
AU2003213606A1 (en) 2002-03-01 2003-10-13 Exelixis, Inc. MP53s AS MODIFIERS OF THE p53 PATHWAY AND METHODS OF USE
US20090162382A1 (en) 2002-03-01 2009-06-25 Bernett Matthew J Optimized ca9 antibodies and methods of using the same
US20050287147A1 (en) 2002-05-15 2005-12-29 Reinhard Ebner Cancer-linked gene as target for chemotherapy
US6660856B2 (en) 2002-03-08 2003-12-09 Kaohsiung Medical University Synthesis of pyrrolo[2,1-c][1,4]benzodiazepine analogues
NZ535425A (en) 2002-03-13 2008-05-30 Biogen Idec Inc Anti-alphavbeta6 antibodies
EP2258712A3 (en) 2002-03-15 2011-05-04 Multicell Immunotherapeutics, Inc. Compositions and Methods to Initiate or Enhance Antibody and Major-histocompatibility Class I or Class II-restricted T Cell Responses by Using Immunomodulatory, Non-coding RNA Motifs
WO2004000997A2 (en) 2002-03-19 2003-12-31 Curagen Corporation Therapeutic polypeptides, nucleic acids encoding same, and methods of use
WO2003081210A2 (en) 2002-03-21 2003-10-02 Sunesis Pharmaceuticals, Inc. Identification of kinase inhibitors
US7193069B2 (en) 2002-03-22 2007-03-20 Research Association For Biotechnology Full-length cDNA
JP2005520566A (ja) 2002-03-22 2005-07-14 バイオジェン・アイデック・エムエイ・インコーポレイテッド Ｃｒｉｐｔｏ特異的抗体
JP2005521429A (ja) 2002-03-25 2005-07-21 ユーエービー リサーチ ファウンデーション Ｆｃレセプターホモログ、試薬およびこれらの使用
AU2003222103A1 (en) 2002-03-28 2003-10-13 Idec Pharmaceuticals Corporation Novel gene targets and ligands that bind thereto for treatment and diagnosis of colon carcinomas
US20030194704A1 (en) 2002-04-03 2003-10-16 Penn Sharron Gaynor Human genome-derived single exon nucleic acid probes useful for gene expression analysis two
US6534660B1 (en) 2002-04-05 2003-03-18 Immunogen, Inc. CC-1065 analog synthesis
US6756397B2 (en) 2002-04-05 2004-06-29 Immunogen, Inc. Prodrugs of CC-1065 analogs
EP1572916A4 (en) 2002-04-05 2007-05-30 Agensys Inc NUCLEIC ACID AND CORRESPONDING PROTEIN WITH THE NAME 98P4B6, SUITABLE FOR THE TREATMENT AND DETECTION OF CANCER
EP2289942B1 (en) 2002-04-10 2013-07-31 Genentech, Inc. Anti-HER2 antibody variants
US20040101874A1 (en) 2002-04-12 2004-05-27 Mitokor Inc. Targets for therapeutic intervention identified in the mitochondrial proteome
JP2005536190A (ja) 2002-04-16 2005-12-02 ジェネンテック・インコーポレーテッド 腫瘍の診断と治療のための組成物と方法
WO2003089904A2 (en) 2002-04-17 2003-10-30 Baylor College Of Medicine Aib1 as a prognostic marker and predictor of resistance to encocrine therapy
SG187991A1 (en) 2002-05-02 2013-03-28 Wyeth Corp Calicheamicin derivative-carrier conjugates
AU2003228869A1 (en) 2002-05-03 2003-11-17 Incyte Corporation Transporters and ion channels
US6596757B1 (en) 2002-05-14 2003-07-22 Immunogen Inc. Cytotoxic agents comprising polyethylene glycol-containing taxanes and their therapeutic use
WO2003101388A2 (en) 2002-05-30 2003-12-11 Bristol-Myers Squibb Company Human solute carrier family 7 member 11 (hslc7a11)
AU2003239969A1 (en) 2002-06-04 2003-12-19 Avalon Pharmaceuticals, Inc. Cancer-linked gene as target for chemotherapy
WO2003101283A2 (en) 2002-06-04 2003-12-11 Incyte Corporation Diagnostics markers for lung cancer
AU2003242633A1 (en) 2002-06-06 2003-12-22 Molecular Engines Laboratories Dudulin genes, non-human animal model: uses in human hematological disease
EP1513934B1 (en) 2002-06-06 2011-03-02 Oncotherapy Science, Inc. Genes and polypeptides relating to human colon cancers
CA2488629A1 (en) 2002-06-07 2003-12-18 Avalon Pharmaceuticals, Inc Cancer-linked gene as target for chemotherapy
US7767803B2 (en) 2002-06-18 2010-08-03 Archemix Corp. Stabilized aptamers to PSMA and their use as prostate cancer therapeutics
US20040022727A1 (en) 2002-06-18 2004-02-05 Martin Stanton Aptamer-toxin molecules and methods for using same
US20040249130A1 (en) 2002-06-18 2004-12-09 Martin Stanton Aptamer-toxin molecules and methods for using same
EP1539218A4 (en) 2002-06-20 2007-08-22 Univ California COMPOSITIONS AND METHODS FOR MODULATING LYMPHOCYTE ACTIVITY
EP2365004B1 (en) 2002-06-21 2016-01-06 Johns Hopkins University School of Medicine Membrane associated tumor endothelium markers
CA2491017A1 (en) 2002-07-03 2004-01-15 Immunogen, Inc. Antibodies to non-shed muc1 and muc16, and uses thereof
US7776814B2 (en) 2002-07-09 2010-08-17 R&D-Biopharmaceuticals Gmbh Tubulysin conjugates
DK1523493T3 (da) 2002-07-09 2013-12-02 Alexander Doemling Nye tubulysinanaloge
DE10230872A1 (de) 2002-07-09 2004-01-22 Morphochem AG Aktiengesellschaft für kombinatorische Chemie Verfahren zur Herstellung von substituierten Thiazol-2-ylmethylestern
WO2004009622A2 (en) 2002-07-19 2004-01-29 Cellzome Ag Protein complexes of cellular networks underlying the development of cancer and other diseases
WO2004011611A2 (en) 2002-07-25 2004-02-05 Genentech, Inc. Taci antibodies and uses thereof
DK1545613T3 (da) 2002-07-31 2011-11-14 Seattle Genetics Inc Auristatinkonjugater og deres anvendelse til behandling af cancer, en autoimmun sygdom eller en infektiøs sygdom
WO2004015426A1 (en) 2002-08-06 2004-02-19 Bayer Healthcare Ag Diagnostics and therapeutics for diseases associated with human cxc chemokine receptor 5(cxcr5)
JP2004121218A (ja) 2002-08-06 2004-04-22 Jenokkusu Soyaku Kenkyusho:Kk 気管支喘息または慢性閉塞性肺疾患の検査方法
DE60336149D1 (de) 2002-08-16 2011-04-07 Immunogen Inc Vernetzer mit hoher reaktivität und löslichkeit und ihre verwendung bei der herstellung von konjugaten für die gezielte abgabe von kleinmolekularen arzneimitteln
MXPA05001933A (es) 2002-08-19 2005-04-28 Genentech Inc Composiciones y metodos para el diagnostico y tratamiento de tumores.
US20090252738A1 (en) 2002-08-23 2009-10-08 Novartis Vaccines And Diagnostics, Inc. Compositions and methods of therapy for cancers characterized by expression of the tumor-associated antigen mn/ca ix
AU2003278725A1 (en) 2002-08-27 2004-03-19 Bristol-Myers Squibb Company Polynucleotide predictor set for identifying protein tyrosine kinase modulators
WO2004020595A2 (en) 2002-08-29 2004-03-11 Five Prime Therapeutics, Inc. Novel human polypeptides encoded by polynucleotides
AU2002951346A0 (en) 2002-09-05 2002-09-26 Garvan Institute Of Medical Research Diagnosis of ovarian cancer
DE10241152A1 (de) 2002-09-05 2004-03-18 GESELLSCHAFT FüR BIOTECHNOLOGISCHE FORSCHUNG MBH (GBF) Tubulysin-Biosynthese-Gene
AU2003270311A1 (en) 2002-09-06 2004-03-29 Mannkind Corporation Epitope sequences
EP1581648A2 (en) 2002-09-09 2005-10-05 Nura, Inc. G protein coupled receptors and uses thereof
JP2004113151A (ja) 2002-09-27 2004-04-15 Sankyo Co Ltd 癌遺伝子及びその用途
US7541440B2 (en) 2002-09-30 2009-06-02 Immunomedics, Inc. Chimeric, human and humanized anti-granulocyte antibodies and methods of use
CA2500978A1 (en) 2002-10-03 2004-04-15 Mcgill University Antibodies and cyclic peptides which bind cea (carcinoembryonic antigen) and their use as cancer therapeutics
AU2003288918A1 (en) 2002-10-04 2004-05-04 Van Andel Research Institute Molecular sub-classification of kidney tumors and the discovery of new diagnostic markers
CN101972478B (zh) 2002-10-08 2012-10-31 免疫医疗公司 抗体治疗
ZA200503075B (en) 2002-11-07 2006-09-27 Immunogen Inc Anti-CD33 antibodies and method for treatment of acute myeloid leukemia using the same
EP1578940A4 (en) 2002-11-13 2007-12-12 Genentech Inc METHOD AND COMPOSITIONS FOR DYSPLASED DIAGNOSIS
GB0226593D0 (en) 2002-11-14 2002-12-24 Consultants Ltd Compounds
AU2003298650B2 (en) 2002-11-15 2010-03-11 Musc Foundation For Research Development Complement receptor 2 targeted complement modulators
WO2004045553A2 (en) 2002-11-15 2004-06-03 The Board Of Trustees Of The University Of Arkansas Ca125 gene and its use for diagnostic and therapeutic interventions
WO2004046342A2 (en) 2002-11-20 2004-06-03 Biogen Idec Inc. Novel gene targets and ligands that bind thereto for treatment and diagnosis of carcinomas
DE10254439A1 (de) 2002-11-21 2004-06-03 GESELLSCHAFT FüR BIOTECHNOLOGISCHE FORSCHUNG MBH (GBF) Tubulysine, Herstellungsverfahren und Tubulysin-Mittel
EP2410332A1 (en) 2002-11-21 2012-01-25 The University Of Utah Method for identifying purinergic modulators of the olfactory system
US20040253606A1 (en) 2002-11-26 2004-12-16 Protein Design Labs, Inc. Methods of detecting soft tissue sarcoma, compositions and methods of screening for soft tissue sarcoma modulators
EP1581641A4 (en) 2002-12-06 2006-11-15 Diadexus Inc COMPOSITIONS, SPREADING VARIATIONS AND METHODS RELATED TO OVARAL SPECIFIC GENES AND PROTEINS
JP2004198419A (ja) 2002-12-13 2004-07-15 Bayer Healthcare Llc Ｔｉｍｐ１を用いた検出方法
WO2004058171A2 (en) 2002-12-20 2004-07-15 Protein Design Labs, Inc. Antibodies against gpr64 and uses thereof
AU2003299819A1 (en) 2002-12-23 2004-07-22 Human Genome Sciences, Inc. Neutrokine-alpha conjugate, neutrokine-alpha complex, and uses thereof
CA2512536A1 (en) 2003-01-08 2004-07-29 Bristol-Myers Squibb Company Biomarkers and methods for determining sensitivity to epidermal growth factor receptor modulators
US20050227301A1 (en) 2003-01-10 2005-10-13 Polgen Cell cycle progression proteins
WO2004063355A2 (en) 2003-01-10 2004-07-29 Protein Design Labs, Inc. Novel methods of diagnosis of metastatic cancer, compositions and methods of screening for modulators of matastatic cancer
WO2004065577A2 (en) 2003-01-14 2004-08-05 Bristol-Myers Squibb Company Polynucleotides and polypeptides associated with the nf-kb pathway
EP1583821A4 (en) 2003-01-15 2007-07-18 Millennium Pharm Inc METHODS AND COMPOSITIONS FOR TREATING UROLOGICAL DISORDERS USING GENES 44390, 54181, 211, 5687, 884, 1405, 636, 4421, 5410, 30905, 2045, 16405, 18560, 2047, 33751, 52872, 14063, 20739, 32544, 43239, 44373, 51164, 53010, 16852, 1587, 2207, 22245, 2387, 52908, 69112, 14990, 18547, 115, 579
EP2336189A1 (en) 2003-01-28 2011-06-22 Proscan RX Pharma Prostate cancer diagnosis and treatment
MXPA05008222A (es) 2003-01-31 2005-10-05 Immunomedics Inc Metodos y composiciones para administrar agentes terapeuticos y de diagnostico.
BRPI0407446A (pt) 2003-02-13 2006-01-31 Pharmacia Corp Anticorpos para c-met para o tratamento de cânceres
AU2004213432A1 (en) 2003-02-14 2004-09-02 Sagres Discovery, Inc. Novel therapeutic targets in cancer
JP5356648B2 (ja) 2003-02-20 2013-12-04 シアトル ジェネティックス， インコーポレイテッド 抗ｃｄ７０抗体−医薬結合体、ならびに癌および免疫障害の処置のためのそれらの使用
US20030224411A1 (en) 2003-03-13 2003-12-04 Stanton Lawrence W. Genes that are up- or down-regulated during differentiation of human embryonic stem cells
US6800622B1 (en) 2003-03-25 2004-10-05 Council Of Scientific And Industrial Research Pyrene-linked pyrrolo[2,1-c][1,4]benzodiazepine hybrids useful as anti-cancer agents
ATE421967T1 (de) 2003-03-31 2009-02-15 Council Scient Ind Res Nichtvernetzende pyrroloä2,1-cüä1, 4übenzodiazepine als potentielle antitumor- agentien und ihre herstellung
US7015215B2 (en) 2003-03-31 2006-03-21 Council Of Scientific And Industrial Research Pyrrolo[2,1-c][1,4] benzodiazepines compounds and process thereof
US8088387B2 (en) 2003-10-10 2012-01-03 Immunogen Inc. Method of targeting specific cell populations using cell-binding agent maytansinoid conjugates linked via a non-cleavable linker, said conjugates, and methods of making said conjugates
US7276497B2 (en) 2003-05-20 2007-10-02 Immunogen Inc. Cytotoxic agents comprising new maytansinoids
WO2005001486A1 (en) 2003-06-06 2005-01-06 Genentech, Inc. Modulating the interaction between hgf beta chain and c-met
KR20130065723A (ko) 2003-06-27 2013-06-19 암젠 프레몬트 인코포레이티드 상피 성장 인자 수용체의 결실 돌연변이체 지향 항체 및 그 용도
CA2534639C (en) 2003-07-31 2013-07-30 Immunomedics, Inc. Anti-cd19 antibodies
US7902338B2 (en) 2003-07-31 2011-03-08 Immunomedics, Inc. Anti-CD19 antibodies
HN2004000285A (es) 2003-08-04 2006-04-27 Pfizer Prod Inc ANTICUERPOS DIRIGIDOS A c-MET
GB0321295D0 (en) 2003-09-11 2003-10-15 Spirogen Ltd Synthesis of protected pyrrolobenzodiazepines
WO2005040170A2 (en) 2003-10-22 2005-05-06 Government Of The United States Of America, Represented By The Secretary, Department Of Health And Human Services Pyrrolobenzodiazepine derivatives, compositions comprising the same and methods related thereto
EP2478912B1 (en) 2003-11-06 2016-08-31 Seattle Genetics, Inc. Auristatin conjugates with anti-HER2 or anti-CD22 antibodies and their use in therapy
CA2548282A1 (en) 2003-12-11 2005-06-30 Genentech, Inc. Methods and compositions for inhibiting c-met dimerization and activation
CA2556752C (en) 2004-02-23 2016-02-02 Genentech, Inc. Heterocyclic self-immolative linkers and conjugates
CA2556025A1 (en) 2004-02-23 2005-09-09 Sugen, Inc. Method of treating abnormal cell growth using c-met and m-tor inhibitors
GB0404577D0 (en) 2004-03-01 2004-04-07 Spirogen Ltd Pyrrolobenzodiazepines
WO2005085251A1 (en) 2004-03-01 2005-09-15 Spirogen Limited 11-hydroxy-5h-pyrrolo[2,1-c][1,4]benzodiazepin-5-one derivatives as key intermediates for the preparation of c2 substituted pyrrolobenzodiazepines
JP4942643B2 (ja) 2004-03-02 2012-05-30 シアトル ジェネティックス， インコーポレイテッド 部分的に付加された抗体およびそれらの結合体化方法
WO2005085260A1 (en) 2004-03-09 2005-09-15 Spirogen Limited Pyrrolobenzodiazepines
US7056913B2 (en) 2004-03-30 2006-06-06 Council Of Scientific And Industrial Research C8—linked pyrrolo[2,1-c][1,4]benzodiazepine-acridone/acridine hybrids
US7189710B2 (en) 2004-03-30 2007-03-13 Council Of Scientific And Industrial Research C2-fluoro pyrrolo [2,1−c][1,4]benzodiazepine dimers
US6951853B1 (en) 2004-03-30 2005-10-04 Council Of Scientific And Industrial Research Process for preparing pyrrolo[2, 1-c] [1,4] benzodiazepine hybrids
FR2869231B1 (fr) 2004-04-27 2008-03-14 Sod Conseils Rech Applic Composition therapeutique contenant au moins un derive de la pyrrolobenzodiazepine et la fludarabine
CN1942459A (zh) 2004-05-14 2007-04-04 免疫基因公司 浆果赤霉素ⅲ化合物的简易合成方法
DE102004030227A1 (de) 2004-06-23 2006-01-26 Dömling, Alexander, Dr. Wirkstoffe mit antiangiogenetischen Eigenschaften
US6979684B1 (en) 2004-06-30 2005-12-27 Council Of Scientific And Industrial Research Pyrrolo[2,1-c][1,4]benzodiazepine-napthalimide conjugates linked through piperazine moiety and process for preparation thereof
KR101205289B1 (ko) 2004-07-02 2012-11-27 빌렉스 아게 Ｇ-250 발현 종양의 개선된 보조제 요법
WO2006091231A2 (en) 2004-07-21 2006-08-31 Ambrx, Inc. Biosynthetic polypeptides utilizing non-naturally encoded amino acids
DK1771482T3 (da) 2004-07-22 2014-10-20 Genentech Inc HER2-antistofsammensætning
CA2575402A1 (en) 2004-08-05 2006-02-09 Genentech, Inc. Humanized anti-cmet antagonists
WO2006033913A2 (en) 2004-09-16 2006-03-30 Synta Pharmaceuticals Corp. Bis (thio-hydrazide amides) for treament of hyperplasia
DK1791565T3 (en) 2004-09-23 2016-08-01 Genentech Inc Cysteingensplejsede antibodies and conjugates
WO2006044643A2 (en) 2004-10-15 2006-04-27 Seattle Genetics, Inc. Anti-cd70 antibody and its use for the treatment and prevention of cancer and immune disorders
JO3000B1 (ar) 2004-10-20 2016-09-05 Genentech Inc مركبات أجسام مضادة .
AU2005304031B2 (en) 2004-11-09 2009-04-23 Philogen Spa Antibodies against tenascin-C
US7892827B2 (en) 2004-11-26 2011-02-22 Pieris Ag Compound with affinity for the cytotoxic T lymphocyte-associated antigen (CTLA-4)
EP1661584A1 (en) 2004-11-26 2006-05-31 Heinz Dr. Faulstich Use of conjugates of amatoxins and phallotoxins with macromolecules for cancer and inflammation therapy
DE602005022928D1 (de) 2004-11-30 2010-09-23 Abgenix Inc Antikörper gegen gpnmb und ihre verwendungen
EP1688415A1 (en) 2004-12-07 2006-08-09 Aventis Pharma S.A. Cytotoxic agents comprising new C-2 modified taxanes
EP1669358A1 (en) 2004-12-07 2006-06-14 Aventis Pharma S.A. Cytotoxic agents comprising new taxanes
EP2399893B1 (en) 2004-12-22 2018-08-15 Ambrx, Inc. Compositions containing, methods involving, and uses of non-natural amino acids and polypeptides
US7173026B2 (en) 2004-12-27 2007-02-06 Council Of Scientific And Industrial Research Pyrrolo [2,1-c][1,4]benzodiazepine-anthraquinone conjugates useful as antitumour agents
US7301019B2 (en) 2005-01-21 2007-11-27 Immunogen, Inc. Method for the preparation of maytansinoid esters
CN101160324B (zh) 2005-02-18 2013-04-24 米德列斯公司 缺乏岩藻糖残基的抗前列腺特异性膜抗原(psma)单克隆抗体
US7691848B2 (en) 2005-03-02 2010-04-06 Wyeth Pyrrolobenzodiazepine arylcarboxamides and derivatives thereof as follicle-stimulating hormone receptor antagonists
EP1863828A4 (en) 2005-03-07 2010-10-13 Archemix Corp STABILIZED APTAMERS FOR PSMA AND THEIR USE AS PROSTATE ACID THERAPEUTICS
NZ561211A (en) 2005-03-25 2011-03-31 Genentech Inc Methods and compositions for modulating hyperstabilized C-met
US7612062B2 (en) 2005-04-21 2009-11-03 Spirogen Limited Pyrrolobenzodiazepines
EP1893615A1 (en) 2005-06-09 2008-03-05 Wyeth a Corporation of the State of Delaware Pyrrolobenzodiazepine pyridine carboxamides and derivatives as follicle-stimulating hormone receptor antagonists
CN104072614B (zh) 2005-07-08 2017-04-26 生物基因Ma公司 抗-αvβ6 抗体及其用途
EP1928912A4 (en) 2005-09-07 2010-02-24 Medimmune Inc EPH ANTI-RECEPTOR ANTIBODIES CONJUGATED TO TOXINS
EP1945647B9 (en) 2005-11-08 2012-11-14 Immunogen, Inc. Processes for preparation of maytansinol
DE112005003752T5 (de) 2005-11-10 2008-11-13 Council Of Scientific & Industrial Research Neues Pyrrolo[2,1-c][1,4]benzodiapezinhybrid und ein Verfahren zu dessen Herstellung
DK2339014T3 (en) 2005-11-16 2015-07-20 Ambrx Inc Methods and compositions comprising non-natural amino acids
RS52060B (sr) 2006-01-25 2012-04-30 Sanofi Citotoksični agensi koji sadrže nove derivate tomaimicina
EP1832577A1 (en) 2006-03-07 2007-09-12 Sanofi-Aventis Improved prodrugs of CC-1065 analogs
US8101727B2 (en) 2006-03-30 2012-01-24 Novartis Ag Compositions and methods of use for antibodies of c-Met
RU2429014C2 (ru) 2006-03-31 2011-09-20 Массачусетс Инститьют Оф Текнолоджи Лечение опухолей, экспрессирующих мутантные рецепторы egf
US20080096819A1 (en) 2006-05-02 2008-04-24 Allozyne, Inc. Amino acid substituted molecules
JP5313129B2 (ja) 2006-05-02 2013-10-09 アロザイン， インコーポレイテッド 非天然アミノ酸置換ポリペプチド
CA2652703C (en) 2006-06-07 2018-08-28 Bioalliance C.V. Antibodies recognizing a carbohydrate containing epitope on cd-43 and cea expressed on cancer cells and methods using same
EP2064244B1 (en) 2006-08-03 2019-06-05 MedImmune Limited ANTIBODIES DIRECTED TO alphaVbeta6 AND USES THEREOF
ITMI20061624A1 (it) 2006-08-11 2008-02-12 Bioker Srl Mono-coniugati sito-specifici di g-csf
PL2383297T3 (pl) 2006-08-14 2013-06-28 Xencor Inc Zoptymalizowane przeciwciała ukierunkowane na CD19
WO2008020456A2 (en) 2006-08-14 2008-02-21 Council Of Scientific & Industrial Research Pyrrolo[2,1-c][1,4]benzodiazepine hybrids and a process for the preparation thereof
CA2660799C (en) 2006-08-14 2013-03-19 Council Of Scientific & Industrial Research Pyrrolo[2,1-c][1,4]benzodiazepine hybrids and a process for the preparation thereof
US7985783B2 (en) 2006-09-21 2011-07-26 The Regents Of The University Of California Aldehyde tags, uses thereof in site-specific protein modification
CN101535332A (zh) 2006-10-31 2009-09-16 免疫基因公司 用于提高抗体产量的方法
TWI528976B (zh) 2006-12-13 2016-04-11 斯茹林製藥公司 用於醫物傳遞之以環糊精為基之聚合物
AU2008224988A1 (en) 2007-03-14 2008-09-18 Endocyte, Inc. Binding ligand linked drug delivery conjugates of tubulysins
WO2008125116A2 (en) 2007-03-31 2008-10-23 Aarhus Universitet Upar inhibition
WO2008122039A2 (en) 2007-04-02 2008-10-09 The Government Of The United States Of America, As Represented By The Secretary, Department Of Health And Human Services Selenocysteine mediated hybrid antibody molecules
CL2008001334A1 (es) 2007-05-08 2008-09-22 Genentech Inc Anticuerpo anti-muc16 disenado con cisteina; conjugado que lo comprende; metodo de produccion; formulacion farmaceutica que lo comprende; y su uso para tratar el cancer.
WO2008138561A1 (en) 2007-05-10 2008-11-20 R & D Biopharmaceuticals Gmbh Tubulysine derivatives
CN101784565B (zh) 2007-06-25 2014-12-10 恩多塞特公司 含有亲水性间隔区接头的共轭物
EP2014681A1 (en) 2007-07-12 2009-01-14 Pierre Fabre Medicament Novel antibodies inhibiting c-met dimerization, and uses thereof
SI2019104T1 (sl) 2007-07-19 2013-12-31 Sanofi Citotoksična sredstva, ki obsegajo nove tomaimicinske derivate, in njihova terapevtska uporaba
US8476451B2 (en) 2007-07-20 2013-07-02 The Regents Of The University Of California Tubulysin D analogues
EP2181101A2 (en) 2007-07-20 2010-05-05 Helmholtz-Zentrum für Infektionsforschung GmbH Tubulysin d analogues
PL2187965T3 (pl) 2007-08-17 2020-05-18 Purdue Research Foundation Koniugaty wiążący psma ligand-łącznik i sposoby ich zastosowania
DK2211904T3 (en) 2007-10-19 2016-10-24 Seattle Genetics Inc Cd19-binding agents and uses thereof
CN101909441B (zh) 2007-10-25 2015-05-13 恩多塞特公司 微管蛋白抑制剂及其制备方法
WO2009056342A1 (en) 2007-11-02 2009-05-07 Wilex Ag Binding epitopes for g250 antibody
HUE026846T2 (en) 2007-12-18 2016-08-29 Bioalliance Cv Antibodies to Recognize Carbohydrate-Containing Epitope on CD43 and CEA Expressed on Cancer Cells and Procedures for their Use
WO2009086072A2 (en) 2007-12-21 2009-07-09 Genentech, Inc. Therapy of rituximab-refractory rheumatoid arthritis patients
US9221914B2 (en) 2007-12-26 2015-12-29 Biotest Ag Agents targeting CD138 and uses thereof
CN101965366B (zh) 2007-12-26 2016-04-27 生物测试股份公司 靶向cd138的免疫缀合物及其应用
TWI472339B (zh) 2008-01-30 2015-02-11 Genentech Inc 包含結合至ｈｅｒ２結構域ii之抗體及其酸性變異體的組合物
CN101932598B (zh) 2008-01-30 2016-12-21 皮里斯股份公司 具有对人c-met受体酪氨酸激酶的亲和性的泪脂质运载蛋白的突变蛋白及其获得方法
EP2265613B1 (en) 2008-03-05 2013-07-24 Council Of Scientific & Industrial Research Quinazoline linked pyrrolo[2,1-c][1, 4]benzodiazepine hybrids as potential anticancer agents and process for the preparation thereof
RU2531754C2 (ru) 2008-04-11 2014-10-27 ЭМЕРДЖЕНТ ПРОДАКТ ДИВЕЛОПМЕНТ СИЭТЛ,ЭлЭлСи,US Связывающееся с cd37 иммунотерапевтическое средство и его комбинация с бифункциональным химиотерапевтическим средством
CN102159243B (zh) 2008-07-21 2015-08-19 免疫医疗公司 用于改良的治疗特征的抗体的结构变体
ES3029483T3 (en) 2008-09-17 2025-06-24 Endocyte Inc Folate receptor binding conjugates of antifolates
EP2174947A1 (en) 2008-09-25 2010-04-14 Universität des Saarlandes Bioactive pre-tubulysins and use thereof
EP2361263A1 (en) 2008-10-31 2011-08-31 Abbott Biotherapeutics Corp. Use of anti-cs1 antibodies for treatment of rare lymphomas
WO2010051502A2 (en) 2008-10-31 2010-05-06 Biogen Idec Ma Inc. Light targeting molecules and uses thereof
PA8849001A1 (es) 2008-11-21 2010-06-28 Lilly Co Eli Anticuerpos de c-met
US8545839B2 (en) 2008-12-02 2013-10-01 Pierre Fabre Medicament Anti-c-Met antibody
AR074439A1 (es) 2008-12-02 2011-01-19 Pf Medicament Anticuerpo anti-cmet (receptor c-met)
JP5798489B2 (ja) 2009-01-12 2015-10-21 ストロ バイオファーマ， インコーポレイテッド インビトロタンパク質合成システムを用いて非天然アミノ酸をタンパク質に選択的に導入するためのモノチャージシステム
RU2683325C2 (ru) 2009-02-05 2019-03-28 Иммьюноджен, Инк. Новые производные бензодиазепина
ES3000111T3 (en) 2009-02-13 2025-02-27 Immunomedics Inc Intermediates for preparing conjugates with an intracellularly-cleavable linkage
WO2010096394A2 (en) 2009-02-17 2010-08-26 Redwood Biosciences, Inc. Aldehyde-tagged protein-based drug carriers and methods of use
CA2761120A1 (en) 2009-05-06 2010-11-11 Biotest Ag Uses of immunoconjugates targeting cd138
EP2427490A1 (en) 2009-05-07 2012-03-14 Novartis AG Compositions and methods of use for binding molecules to dickkopf-1 or dickkopf-4 or both
JP5310337B2 (ja) 2009-07-15 2013-10-09 セイコーエプソン株式会社 流体噴射装置、流体噴射装置における流体噴射ヘッド制御方法及び流体噴射ヘッド用の駆動波形生成装置
IT1394860B1 (it) 2009-07-22 2012-07-20 Kemotech S R L Composti farmaceutici
US8394922B2 (en) 2009-08-03 2013-03-12 Medarex, Inc. Antiproliferative compounds, conjugates thereof, methods therefor, and uses thereof
EP2478110B1 (en) 2009-09-16 2016-01-06 Immunomedics, Inc. Class i anti-cea antibodies and uses thereof
US20110028129A1 (en) 2009-10-13 2011-02-03 Hutchison James W Proximity Triggered Profile-Based Wireless Matching
KR101671378B1 (ko) 2009-10-30 2016-11-01 삼성전자 주식회사 ｃ-Ｍｅｔ에 특이적으로 결합하는 항체 및 그의 용도
EP2513148B1 (en) 2009-12-16 2016-08-31 AbbVie Biotherapeutics Inc. Anti-her2 antibodies and their uses
US8609095B2 (en) 2010-03-04 2013-12-17 Symphogen A/S Anti-HER2 antibodies and compositions
JP5532333B2 (ja) 2010-10-21 2014-06-25 株式会社パトライト 壁に取り付け可能な機器
EP2697218B1 (en) * 2011-04-13 2016-05-25 Janssen Pharmaceutica NV Process for the preparation of compounds useful as inhibitors of sglt2
TWI495644B (zh) 2011-11-11 2015-08-11 Rinat Neuroscience Corp 營養層細胞表面抗原（Ｔｒｏｐ－２）之專一性抗體類及彼等之用途
EP2793947B1 (en) 2011-12-23 2021-02-03 Innate Pharma Enzymatic conjugation of polypeptides
GB201210770D0 (en) 2012-06-18 2012-08-01 Polytherics Ltd Novel conjugation reagents
ES2781523T3 (es) 2012-07-12 2020-09-02 Hangzhou Dac Biotech Co Ltd Conjugados de moléculas de unión celular con agentes citotóxicos
US20150218220A1 (en) 2012-09-12 2015-08-06 Brian Alan MENDELSOHN Amatoxin derivatives and cell-permeable conjugates thereof as inhibitors of rna polymerase
ES2773710T3 (es) 2012-10-11 2020-07-14 Daiichi Sankyo Co Ltd Enlazadores para conjugados de anticuerpo - fármaco
NO2789793T3 (https=) 2012-10-24 2018-01-27
US9310374B2 (en) 2012-11-16 2016-04-12 Redwood Bioscience, Inc. Hydrazinyl-indole compounds and methods for producing a conjugate
US9243058B2 (en) * 2012-12-07 2016-01-26 Amgen, Inc. BCMA antigen binding proteins
US9701753B2 (en) 2013-03-11 2017-07-11 Genzyme Corporation Hyperglycosylated binding polypeptides
JP6114176B2 (ja) * 2013-03-15 2017-04-12 信越化学工業株式会社 対称ハイパーブランチ型シリコーン変性重合性化合物、及びそのモジュール化した製造方法
WO2015155345A1 (en) 2014-04-11 2015-10-15 Medimmune Limited Antibodies and antibody-drug conjugates
GB201406767D0 (en) 2014-04-15 2014-05-28 Cancer Rec Tech Ltd Humanized anti-Tn-MUC1 antibodies anf their conjugates
CN116059395A (zh) * 2015-06-29 2023-05-05 第一三共株式会社 用于选择性制造抗体-药物缀合物的方法
EP3317246A4 (en) 2015-07-04 2019-02-27 Suzhou M-conj Biotech Co., Ltd. SPECIFIC CONJUGATION OF A CELL BINDING MOLECULE
NZ739830A (en) * 2015-07-12 2021-12-24 Hangzhou Dac Biotech Co Ltd Bridge linkers for conjugation of cell-binding molecules
JP6817288B2 (ja) * 2015-08-10 2021-01-20 ハンジョウ ディーエーシー バイオテック シーオー．，エルティディ．Ｈａｎｇｚｈｏｕ Ｄａｃ Ｂｉｏｔｅｃｈ Ｃｏ．，Ｌｔｄ． 新規な連結体及び生体分子と薬物との特異的共役におけるその使用
JP2019505476A (ja) * 2015-12-01 2019-02-28 グラクソスミスクライン、インテレクチュアル、プロパティー、ディベロップメント、リミテッドＧｌａｘｏｓｍｉｔｈｋｌｉｎｅ Ｉｎｔｅｌｌｅｃｔｕａｌ Ｐｒｏｐｅｒｔｙ Ｄｅｖｅｌｏｐｍｅｎｔ Ｌｉｍｉｔｅｄ 組合せ処置およびその方法
US11129910B2 (en) * 2016-02-04 2021-09-28 Hangzhou Dac Biotech Co., Ltd. Specific conjugation linkers, specific immunoconjugates thereof, methods of making and uses such conjugates thereof
PE20231050A1 (es) 2016-03-02 2023-07-11 Eisai Randd Man Co Ltd Conjugados de anticuerpo y farmaco basados en eribulina y metodos para su uso
KR20220150408A (ko) * 2016-11-14 2022-11-10 항저우 디에이씨 바이오테크 씨오, 엘티디 결합 링커, 그러한 결합 링커를 함유하는 세포 결합 분자-약물 결합체, 링커를 갖는 그러한 결합체의 제조 및 사용
KR20210122319A (ko) 2017-04-06 2021-10-08 항저우 디에이씨 바이오테크 씨오, 엘티디 비스-링키지를 사용한 세포독성 약물의 접합
MA51447A (fr) * 2017-08-01 2020-06-10 Medimmune Llc Conjugué anticorps monoclonal-médicament dirigé contre bcma
EP3621652A4 (en) * 2017-12-31 2021-01-13 Hangzhou DAC Biotech Co, Ltd CONJUGATE OF ANALOGUE OF TUBULYSIN WITH BRANCHED LINKS
KR102870868B1 (ko) * 2018-06-01 2025-10-15 노파르티스 아게 Bcma에 대한 결합 분자 및 이의 용도
IL282182B2 (en) 2018-10-12 2025-09-01 Hangzhou Dac Biotech Co Ltd Cytotoxic drug conjugates with a bis linkage
US11478553B2 (en) * 2019-02-15 2022-10-25 Wuxi Biologies Ireland Limited Process for preparing antibody-drug conjugates with improved homogeneity
JP2022531001A (ja) * 2019-05-03 2022-07-05 セルジーン コーポレーション 抗ｂｃｍａ抗体コンジュゲート、そのコンジュゲートを含む組成物ならびにそのコンジュゲートの製造および使用方法
WO2020227105A1 (en) * 2019-05-03 2020-11-12 Sutro Biopharma, Inc. Anti-bcma antibody conjugates
CN110229232B (zh) * 2019-06-19 2020-05-19 北京智仁美博生物科技有限公司 双特异性抗体及其用途
CN112168978B (zh) * 2019-07-03 2022-01-11 北京大学 一种抗体偶联药物及其药物组合物与应用
CN112409482B (zh) * 2019-08-20 2022-08-26 杭州尚健生物技术有限公司 Bcma抗体
EP3862023A1 (en) * 2020-02-05 2021-08-11 Hangzhou DAC Biotech Co, Ltd Conjugates of cell-binding molecules with cytotoxic agents
EP4426727A2 (en) 2021-11-03 2024-09-11 Hangzhou Dac Biotech Co., Ltd. Specific conjugation of an antibody

Cited By (46)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US12122826B2 (en) 2016-04-27 2024-10-22 Abbvie Inc. Methods of treatment of diseases in which IL-13 activity is detrimental using anti-IL-13 antibodies
US12129294B2 (en) 2016-04-27 2024-10-29 Abbvie Inc. Methods of treatment of diseases in which IL-13 activity is detrimental using anti-IL-13 antibodies
US12331130B2 (en) 2019-12-17 2025-06-17 The Board Of Regents Of The University Of Texas System DDR1 antibodies and uses thereof
US12202857B2 (en) 2020-05-08 2025-01-21 Waters Technologies Corporation Methods for heat-assisted enzyme digestion
US12411064B2 (en) 2020-05-08 2025-09-09 Waters Technologies Corporation Methods for heat-assisted enzyme digestion
JP2024519140A (ja) * 2021-05-27 2024-05-08 サイムワークス・ビーシー・インコーポレイテッド カンプトテシン類似体、複合体及び使用の方法
EP4347601A4 (en) * 2021-05-27 2025-05-21 Zymeworks BC Inc. CAMPTOTHECIN ANALOGUES, CONJUGATES AND METHODS OF USE
WO2023078273A1 (en) 2021-11-03 2023-05-11 Hangzhou Dac Biotech Co., Ltd. Specific conjugation for an antibody-drug conjugate
US12433952B2 (en) 2022-03-25 2025-10-07 Zymeworks Bc Inc. Antibody-drug conjugates targeting folate receptor alpha and methods of use
WO2023201268A1 (en) 2022-04-13 2023-10-19 Gilead Sciences, Inc. Combination therapy for treating tumor antigen expressing cancers
WO2023201267A1 (en) 2022-04-13 2023-10-19 Gilead Sciences, Inc. Combination therapy for treating trop-2 expressing cancers
JP2025513310A (ja) * 2022-04-29 2025-04-24 シャンハイ ミクウルク ファーマシューティカル シーオー．エルティーディー カンプトテシン類似体のリガンド－薬物複合体、中間体、その製造方法、医薬組成物及びその用途
WO2023222108A1 (zh) * 2022-05-20 2023-11-23 上海迈晋生物医药科技有限公司 抗体-药物偶联物的制备方法
WO2023230620A1 (en) * 2022-05-27 2023-11-30 Sonnet BioTherapeutics, Inc. Il-12-albumin-binding domain fusion protein formulations and methods of use thereof
CN115181076A (zh) * 2022-06-24 2022-10-14 北京丹大生物技术有限公司 一种用于检测阿立哌唑和脱氢阿立哌唑浓度的半抗原、抗原、细胞株、抗体、试剂和试剂盒
CN115181076B (zh) * 2022-06-24 2023-03-24 北京丹大生物技术有限公司 一种用于检测阿立哌唑和脱氢阿立哌唑浓度的半抗原、抗原、细胞株、抗体、试剂和试剂盒
WO2024020379A3 (en) * 2022-07-19 2024-03-14 Praesidia Biotherapeutics Inc. Prodrugs, prodrug compositions and related methods
JP2025524170A (ja) * 2022-07-29 2025-07-25 杭州愛科瑞思生物医薬有限公司 カンプトテシン誘導体及びその調製方法ならびに使用
EP4541797A4 (en) * 2022-07-29 2026-03-18 Hangzhou Adcoris Biopharma Co Ltd CAMPTOTHECINE DERIVATIVE, ITS PREPARATION PROCESS AND ITS USE
WO2024041545A1 (en) * 2022-08-22 2024-02-29 Suzhou Bioreinno Biotechnology Limited Company A novel thiol reductant, preparation method and use thereof
WO2024041543A1 (en) * 2022-08-22 2024-02-29 Suzhou Bioreinno Biotechnology Limited Company A method of preparing an antibody with thiol group site-specific modifications and use of tcep
JP2025527006A (ja) * 2022-08-22 2025-08-15 スージョウ バイオレイノ バイオテクノロジー リミテッド カンパニー 部位特異的修飾を有する抗体の調製方法
WO2024041544A1 (en) * 2022-08-22 2024-02-29 Suzhou Bioreinno Biotechnology Limited Company A method of preparing an antibody with site-specific modifications
EP4577552A4 (en) * 2022-08-22 2026-02-25 Suzhou Bioreinno Biotechnology Ltd Company NEW THIOL REDUCER, PROCESS AND ITS USE
JP2025527005A (ja) * 2022-08-22 2025-08-15 スージョウ バイオレイノ バイオテクノロジー リミテッド カンパニー 新規なチオール還元剤、その製造方法およびその用途
WO2024041541A1 (en) * 2022-08-22 2024-02-29 Suzhou Bioreinno Biotechnology Limited Company A novel thiol reductant, method and use thereof
WO2024054673A1 (en) * 2022-09-09 2024-03-14 Texas Tech University System Listeria monocytogenes as a vector for tumor-specific delivery of chemotherapeutic agents
WO2024051787A1 (zh) * 2022-09-09 2024-03-14 北京惠之衡生物科技有限公司 一种长效酰化胰岛素衍生物及其应用
WO2024097812A1 (en) 2022-11-04 2024-05-10 Gilead Sciences, Inc. Therapy for treating bladder cancer
WO2024225837A1 (ko) * 2023-04-28 2024-10-31 주식회사 에이비켐바이오 말레이미드 함유 링커-페이로드 접합체 및 이를 포함하는 신규한 항체-약물 중합체
WO2025241452A1 (en) * 2023-05-24 2025-11-27 Hangzhou Seehe Biotechnology Co., Ltd An antibody-drug conjugate containing an extra functional compound for enhanced treatment of refractory diseases
WO2025241603A1 (en) * 2023-05-24 2025-11-27 Hangzhou Seehe Biotechnology Co., Ltd An antibody drug conjugate with opened rings of thiosuccinimides, its preparation and application thereof
WO2024255873A1 (zh) * 2023-06-14 2024-12-19 江苏迈威康新药研发有限公司 一种制备抗体药物偶联物的方法
CN116879425A (zh) * 2023-06-26 2023-10-13 盛禾(中国)生物制药有限公司 一种双特异性抗体蛋白纯度的检测方法
WO2025002310A1 (zh) * 2023-06-29 2025-01-02 石药集团巨石生物制药有限公司 抗体-药物偶联物及其用途
CN116773826A (zh) * 2023-08-21 2023-09-19 迪亚莱博（张家港）生物科技有限公司 一种用于检测抗蛋白酶3抗体的胶乳比浊生化试剂盒
CN116773826B (zh) * 2023-08-21 2023-11-17 迪亚莱博（张家港）生物科技有限公司 一种用于检测抗蛋白酶3抗体的胶乳比浊生化试剂盒
CN117257977A (zh) * 2023-11-07 2023-12-22 正大天晴药业集团南京顺欣制药有限公司 用于制备抗体药物偶联物的方法
CN117304790A (zh) * 2023-11-27 2023-12-29 石狮佳南热熔胶有限公司 水性环保涂料及水性皮革
CN117304790B (zh) * 2023-11-27 2024-02-09 石狮佳南热熔胶有限公司 水性环保涂料及水性皮革
WO2025119322A1 (en) * 2023-12-07 2025-06-12 Wuxi Biologics (Shanghai) Co., Ltd. Gpc3 antibody-drug conjugates and use thereof
US12286463B1 (en) 2023-12-29 2025-04-29 Sonnet BioTherapeutics, Inc. Interleukin 18 (IL-18) variants and fusion proteins comprising same
US12134635B1 (en) 2023-12-29 2024-11-05 Sonnet BioTherapeutics, Inc. Interleukin 18 (IL-18) variants and fusion proteins comprising same
WO2025151423A1 (en) * 2024-01-08 2025-07-17 OncoC4, Inc. Anti-cd24 antibody drug conjugates and uses thereof
WO2025207442A1 (en) * 2024-03-23 2025-10-02 Ranok Therapeutics (Hangzhou) Co. Ltd. Methods and compositions for targeted protein degradation
WO2026074296A1 (en) * 2024-10-03 2026-04-09 Iksuda Therapeutics Limited Linkers and protein and peptide conjugates incorporating the same

Also Published As

Publication number Publication date
KR20240095316A (ko) 2024-06-25
ZA202403371B (en) 2025-06-25
AU2022381163A1 (en) 2024-06-13
WO2023078021A1 (en) 2023-05-11
CA3236754A1 (en) 2023-05-11
CN117980327A (zh) 2024-05-03
CN118434765A (zh) 2024-08-02
JP2024541059A (ja) 2024-11-06
TW202334217A (zh) 2023-09-01
EP4426741A1 (en) 2024-09-11
TW202506738A (zh) 2025-02-16
EP4426741A4 (en) 2026-03-18
EP4426727A2 (en) 2024-09-11
AU2022383265A1 (en) 2024-06-13
JP2024542073A (ja) 2024-11-13
KR20240095442A (ko) 2024-06-25
WO2022078524A3 (en) 2022-08-25
CA3236930A1 (en) 2022-04-21
WO2023078273A1 (en) 2023-05-11
JP2024541058A (ja) 2024-11-06
CN118215676A (zh) 2024-06-18
TWI864472B (zh) 2024-12-01
WO2022078524A4 (en) 2022-12-08
EP4426729A1 (en) 2024-09-11
AU2021362997A1 (en) 2024-05-16
CA3236852A1 (en) 2023-05-11
TW202430562A (zh) 2024-08-01

Similar Documents

Publication Publication Date Title
WO2022078524A2 (en) Specific conjugation of an antibody
US20230001001A1 (en) Conjugation of a cytotoxic drug with bis-linkage
WO2020257998A1 (en) A conjugate of a cytotoxic agent to a cell binding molecule with branched linkers
US11998584B2 (en) Conjugate of a tubulysin analog with branched linkers
WO2021212638A1 (en) Conjugates of a cell-binding molecule with camptothecin analogs
WO2020073345A1 (en) Conjugation linkers containing 2,3-diaminosuccinyl group
WO2020006722A1 (en) Cross-linked pyrrolobenzodiazepine dimer (pbd) derivative and its conjugates
WO2020155017A1 (en) A conjugate of an amanita toxin with branched linkers
WO2025241603A1 (en) An antibody drug conjugate with opened rings of thiosuccinimides, its preparation and application thereof

Legal Events

Date Code Title Description
WWE Wipo information: entry into national phase

Ref document number: 202180099689.6

Country of ref document: CN

WWE Wipo information: entry into national phase

Ref document number: 810403

Country of ref document: NZ

Ref document number: AU2021362997

Country of ref document: AU

ENP Entry into the national phase

Ref document number: 3236930

Country of ref document: CA

ENP Entry into the national phase

Ref document number: 2024526535

Country of ref document: JP

Kind code of ref document: A

ENP Entry into the national phase

Ref document number: 2021362997

Country of ref document: AU

Date of ref document: 20211103

Kind code of ref document: A

WWE Wipo information: entry into national phase

Ref document number: 11202402716T

Country of ref document: SG

WWE Wipo information: entry into national phase

Ref document number: 2021879565

Country of ref document: EP

NENP Non-entry into the national phase

Ref country code: DE

121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 21879565

Country of ref document: EP

Kind code of ref document: A2

ENP Entry into the national phase

Ref document number: 2021879565

Country of ref document: EP

Effective date: 20240603