WO2001066155A2 - Complexes et conjugat d'actinium-225 pour radio-immunotherapie - Google Patents

Complexes et conjugat d'actinium-225 pour radio-immunotherapie Download PDF

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WO2001066155A2
WO2001066155A2 PCT/US2001/005927 US0105927W WO0166155A2 WO 2001066155 A2 WO2001066155 A2 WO 2001066155A2 US 0105927 W US0105927 W US 0105927W WO 0166155 A2 WO0166155 A2 WO 0166155A2
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hydrogen
independently
group
proviso
alkyl
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PCT/US2001/005927
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WO2001066155A8 (fr
WO2001066155A3 (fr
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Dangshe Ma
Michael R. Mcdevitt
David A. Scheinberg
Jaime Simon
Garry E. Kiefer
R. Keith Frank
Gyongyi Gulyas
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Dangshe Ma
Mcdevitt Michael R
Scheinberg David A
Jaime Simon
Kiefer Garry E
Frank R Keith
Gyongyi Gulyas
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Application filed by Dangshe Ma, Mcdevitt Michael R, Scheinberg David A, Jaime Simon, Kiefer Garry E, Frank R Keith, Gyongyi Gulyas filed Critical Dangshe Ma
Priority to HU0203743A priority Critical patent/HUP0203743A2/hu
Priority to AU49069/01A priority patent/AU4906901A/en
Priority to EP01922246A priority patent/EP1227849A2/fr
Priority to CA2381123A priority patent/CA2381123C/fr
Publication of WO2001066155A2 publication Critical patent/WO2001066155A2/fr
Publication of WO2001066155A8 publication Critical patent/WO2001066155A8/fr
Publication of WO2001066155A3 publication Critical patent/WO2001066155A3/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
    • A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
    • A61K51/04—Organic compounds
    • A61K51/08—Peptides, e.g. proteins, carriers being peptides, polyamino acids, proteins
    • A61K51/10—Antibodies or immunoglobulins; Fragments thereof, the carrier being an antibody, an immunoglobulin or a fragment thereof, e.g. a camelised human single domain antibody or the Fc fragment of an antibody
    • A61K51/1093—Antibodies or immunoglobulins; Fragments thereof, the carrier being an antibody, an immunoglobulin or a fragment thereof, e.g. a camelised human single domain antibody or the Fc fragment of an antibody conjugates with carriers being antibodies
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
    • A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
    • A61K51/04—Organic compounds
    • A61K51/0474—Organic compounds complexes or complex-forming compounds, i.e. wherein a radioactive metal (e.g. 111In3+) is complexed or chelated by, e.g. a N2S2, N3S, NS3, N4 chelating group
    • A61K51/0482—Organic compounds complexes or complex-forming compounds, i.e. wherein a radioactive metal (e.g. 111In3+) is complexed or chelated by, e.g. a N2S2, N3S, NS3, N4 chelating group chelates from cyclic ligands, e.g. DOTA
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
    • A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
    • A61K51/04—Organic compounds
    • A61K51/0497—Organic compounds conjugates with a carrier being an organic compounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
    • A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
    • A61K51/04—Organic compounds
    • A61K51/08—Peptides, e.g. proteins, carriers being peptides, polyamino acids, proteins
    • A61K51/10—Antibodies or immunoglobulins; Fragments thereof, the carrier being an antibody, an immunoglobulin or a fragment thereof, e.g. a camelised human single domain antibody or the Fc fragment of an antibody
    • A61K51/1027—Antibodies or immunoglobulins; Fragments thereof, the carrier being an antibody, an immunoglobulin or a fragment thereof, e.g. a camelised human single domain antibody or the Fc fragment of an antibody against receptors, cell-surface antigens or cell-surface determinants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • This invention relates to actinium-225 ( 25 Ac) complexes with fuctionalized chelants, their conjugates and their use for radioimmunotherapy.
  • radionuclides co plexed with suitable chelants as well as their conjugates (that is, such complexes covalently attached to a biologically active carrier, for example, protein) for diagnosis of cancer and/or therapeutic treatment of cancer in mammals is known.
  • biochemically engineered molecules provide the tumor specificity and the radioisotope provides potent cytotoxicity . See, for example, U.S. Patent Nos. 4,897,254; 5,342,925; 5,435,990; 5,652,361; 5,696,239; and 5,756,065.
  • alpha particles would allow extraordinary potent, single cell-specific killing with minimal toxicity to normal cells or the patient.
  • the use of alpha particles as an alternative to more traditional classes of radiation is derived from the particle's kinetic characteristics and the radioactive half-life of their source isotope, as well as from the properties of the target-selective carrier moiety for the source isotope.
  • alpha emitting radionuclides are highly desirable for the following reasons: (a) a single atom can kill a cell making them hundreds to thousands of times more potent than even the most potent toxins or drugs; (b) the range of alpha particles is only about 50 microns, so that adjacent tissues are not harmed; (c) the chelated atoms on humanized antibodies are unlikely to be immunogenic and can be repeatedly dosed;
  • mice inoculated intraperitoneally with the murine tumor line EL-4 were cured of their ascites after intraperitoneal injection of 150 ⁇ Ci of a 212 Bi-labeled antibody conjugate, see, for example, R. M. Macklis et al, Science, Vol. 240, pp. 1024-1026 (1988).
  • chelating agents such as, for example, 1, 4, 7, 10-tetra- azacyclododecane-1, 4, 7, 10-tetraacetic acid (DOTA) , diethylenetriaminepentaacetic acid (DTPA) , ethylene- diaminetetracetic acid (EDTA), 1, 4, 7 , 10, 13-pentaazacyclo- pentadecane-1, 4, 1 , 10, 13-pentaacetic acid (PEPA) , and 1,4,7,10,13, 16-hexaazacyclohexadecane-l, 4, 7, 10, 13, 16- hexaacetic acid (HEHA) have been complexed with 225 Ac and evaluated in vivo for toxicity and stability. However, the toxicity of these complexes has proved to be still substantial .
  • EDTMP ethylenedia inetetra- ethylenephosphonic acid
  • the study has found that EDTMP, depending on its concentration, reduces the liver uptake. However, the liver uptake of 225 Ac-EDTMP is still substantial and excretion of 225 Ac-EDTMP is poor.
  • the study has also suggested that greater efficacy in endoradionuclide therapy of bone metastasis can be expected with the use of 225 Ac-EDTMP due to the alpha-radiation.
  • conjugates of such 225 Ac complexes with a biological molecule would provide the ttuummoorr ssppeecciiffiicciittyy aarnd the 225 Ac isotope would provide potent cytotoxicity.
  • conjugates Another desirable property of these conjugates includes physiological compatibility which would permit the 225 Ac complex, if separated from its targeting, conjugated biological molecule in vivo, to be soluble in physiological fluids and thus be rapidly eliminated from the body.
  • the present invention is directed to 225 Ac complexes and their conjugates with a biological molecule.
  • the 225 Ac compelexes and conjugates of the present invention are useful for the treatment of cancer in mammals, especially humans .
  • the present invention is directed to 225 AAcc ccoommpplleexxeess ccoommpprriissiiing a functionalized chelant compound of the formula (I)
  • each Q is independently hydrogen or (CHR 5 ) p C0 2 R;
  • Q 1 is hydrogen or (CHR 5 ) w C0 2 R; each R independently is hydrogen, benzyl or C ⁇ -C alkyl; with the proviso that at least two of the sum of Q and Q 1 must be other than hydrogen; each R 5 independently is hydrogen; C3.-C 4 alkyl or (Ci-
  • X and Y are each independently hydrogen or may be taken with an adjacent X and Y to form an additional carbon-carbon bond; n is 0 or 1; m is an integer from 0 to 10 inclusive; p is 1 or 2; r is 0 or 1; w is 0 or 1; with the proviso that n is only 1 when X and/or Y form an additional carbon-carbon bond, and the sum of r and w is 0 or 1; L is a linker/spacer group covalently bonded to, and replaces one hydrogen atom of one of the carbon atoms to which it is joined, said linker/spacer group being represented by the formula
  • the present invention is also directed to a conjugate compriissiinngg tthhee aaffoorreemmeennttiioonneedd 222255 AAcc complex covalently attached to a biological molecule.
  • the present invention also includes formulations having the conjugates of this invention and a pharmaceutically acceptable carrier, especially formulations where the pharmaceutically acceptable carrier is a liquid.
  • the present invention is also directed to a method of therapeutic treatment of a mammal having cancer which comprises administering to said mammal a therapeutically effective amount of the formulation of this invention.
  • the 225 Ac complexes and conjugates of this invention are relatively stable (that is, do not easily dissociate) and some display rapid clearance from the whole body and some non-target organs, such as liver and kidney. Additionally, the alpha-particles emitting 25 Ac complexes and conjugates of this invention are expected to have several advantages over beta particle- emitting cytotoxic agents including higher energy and more potent emissions, less hazardous waste, expected lower effective dose, the potential for outpatient treatment, better retention at the target sites, and higher target to non-target radiation ratios.
  • 225 Ac complex refers to a functionalized chelant compound of formula I complexed with 225 Ac radionuclide .
  • 2 Ac conjugate refers to 225 Ac complex of the present invention that is covalently attached to a biological molecule.
  • the term “mammal” means animals that nurish their young with milk secreted by mammary glands, preferably humans.
  • biological molecule refers to any protein, antibody, antibody fragment, hormone, peptide, growth factor, antigen, hapten or any other carrier which functions in this invention to recognize a specific biological target site.
  • Antibody and antibody fragment refers to any polyclonal, monoclonal, chimeric, human, mammalian, single chains, dimeric and tetrameric antibody or antibody fragment.
  • Such biological molecule when attached to a functionalized complex, serves to carry the attached 225 Ac ion to specific targeted tissues.
  • antibody refers to any polyclonal, monoclonal, chimeric antibody or heteroantibody .
  • the antibodies used in the 225 Ac conjugates of the present invention are monoclonal antibodies having high specificity for the desired cancer cells .
  • Antibodies used in the present invention may be directed against, for example, cancer, tumors, leukemias, autoimune disorders involving cells of the immune system, normal cells that need to be ablated such as bone marrow and prostate tissue, virus infected cells including HIV, mycoplasma, differentiation and other cell membrane antigens, patogen surface antigens and any biologically active molecules .
  • HuM195 anti-CD33
  • CC-11, CC-46,CC-49, CC-49 F(ab') 2 , CC-83, CC-83 F(ab') 2 , and B72.3 Particularly preferred antibody for use in the practice of the present invention is HuM195.
  • Antibody fragment includes Fab fragments and F(ab') 2 fragments, and any portion of an antibody having specificity toward a desired epitope or epitopes .
  • the antibodies which may be used in the 225 Ac conjugates of the present invention can be prepared by techniques well known in the art.
  • pharmaceutically acceptable salt means any salt of a compound of formula (I) which is sufficiently non-toxic to be useful in therapy of mammals.
  • salts which are formed by standard reactions, from both organic and inorganic sources include, for example, sulfuric, hydrochloric, phosphoric, acetic, succinic, citric, lactic, maleic, fumaric, palmitic, cholic, palmoic, mucic, glutamic, d-camphoric, glutaric, glycolic, phthalic, tartaric, formic, lauric, steric, salicylic, methanesulfonic, bensenesulfonic, sorbic, picric, benzoic, cinnamic and other suitable acids.
  • salts formed by standard reactions from both organic and inorganic sources such as ammonium, alkali metal ions, alkaline earth metal ions, and other similar ions.
  • the 225 Ac conjugate may be administered per se or as a component of a pharmaceutically acceptable formulation.
  • the present invention may be practiced with the 225 Ac conjugate being provided in pharmaceutical formulation, both for veterinary and for human medical use.
  • Such pharmaceutical formulations comprise the active agent (the 225 Ac conjugate) together with a physiologically acceptable carrier, excipient or vehicle therefore.
  • the carrier (s) must be physiologically acceptable in the sense of being compatible with the other ingredient (s) in the formulation and not unsuitably deleterious to the recipient thereof.
  • the 225 Ac conjugate is provided in a therapeutically effective amount, as described above, and in a quantity appropriate to achieve the desired dose.
  • the formulations include those suitable for parenteral (including subcutaneous, intramuscular, intraperitoneal, and intravenous), oral, rectal, topical, nasal, or ophthalmic administration.
  • Formulations may be prepared by any methods well known in the art of pharmacy. Such methods include the step of bringing the 225 Ac conjugate into association with a carrier, excipient or vehicle therefore.
  • the formulation may be prepared by uniformly and intimately bringing the 25 Ac conjugate into association with a liquid carrier, a finely divided solid carrier, or both, and then, if necessary, shaping the product into desired formulation.
  • formulations of this invention may further include one or more accessory ingredient (s) selected from diluents, buffers, binders, disintegrants, surface active agents, thickeners, lubricants, preservatives, and the like.
  • a treatment regime might include pretreatment with non-radioactive carrier.
  • Injectable formulations of the present invention may be either in suspensions or solution form.
  • suitable formulations it will be recognized that, in general, the water solubility of the salt is greater than the acid form.
  • a physiologically acceptable carrier Such carriers comprise a suitable solvent, preservatives such as free radical quenching agents, for example, ascorbic acid, benzyl alcohol or any other suitable molecule, if needed, and buffers.
  • suitable solvents include, for example, water, aqueous alcohols, glycols, and phosphonate or carbonate esters. Such aqueous solutions contain no more* than 50 percent of the organic solvent by volume.
  • Injectable suspensions are compositions of the present invention that require a liquid suspending medium, with or without adjuvants, as a carier.
  • the suspending medium can be, for example, aqueous polyvinylpyrrolidone, inert oils such as vegetable oils or highly refined mineral oils, polyols, or aqueous carboxymethylcellulose .
  • Suitable physiologically acceptable adjuvants, if necessary to keep the complex in suspension may be chosen from among thickeners such as carboxymethylcellulose, polyvinylpyrrolidone, gelatin, and the alginates.
  • surfactants are also useful as suspending agents, for example, lecithin, alkylphenol, polyethyleneoxide adducts, naphthalenesulfonates, alkylbenzenesulfonates, and polyoxyethylene sorbitane esters .
  • the terms "functionalized chelant” and "bifunctional chelant” are used interchangeably and refer to compounds which have the dual functionality of sequestering metal ions plus the ability to covalently bind a biological molecule having specificity for tumor cell epitopes or antigens. Such compounds are of great utility for therapeutic and diagnostic applications when they are, for example, complexed with radioactive metal ions and covalently attached to a specific antibody. These types of complexes have been used to carry radioactive metals to tumor cells which are targeted by the specificity of the attached antibody [see, for example, Mears et al . , Anal . Biochem . 142, 68-74 (1984); Krejcarek et al . , Biochem. And Biophys , Res. Comm. 77, 581-585 (1977)].
  • R 2 is selected from the group consisting of hydrogen, nitro, amino, isothiocyanato, semicarbazido, thiosemicarbazido, carboxyl, bromoacetamido and maleimido;
  • R 3 is selected from the group consisting of C ⁇ C 4 alkoxy, -OCH 2 C0 2 H, hydroxy and hydrogen;
  • R 4 is selected from the group consisting of hydrogen, nitro, amino, isothiocyanato, semicarbazido, thiosemicarbazido, carboxyl, bromoacetamido and maleimido; with the proviso that R 2 and R 4 cannot both be hydrogen but one of R 2 and R 4 must be hydrogen; or a pharmaceutically acceptable salt thereof; are preferred functionalized chelants.
  • Preferred functionalized chelant compounds of formula I include also those compounds where Q 1 is hydrogen and L is represented by formula A as shown by formula II:
  • each Q independently is hydrogen or CHR 5 COOR; with the proviso that at least two of Q must be other than hydrogen each R independently is hydrogen benzyl or C 1 -C 4 alkyl; m is integer from 0 to 5 inclusive;
  • R 2 is selected from the group consisting of hydrogen, nitro, amino, isothiocyanato, semicarbazido, thiosemicarbazido, carboxyl, bromoacetamido and maleimido;
  • R 3 is selected from the group consisting of C 1 -C 4 alkoxy, -OCH 2 COOH, hydroxy and hydrogen;
  • R 4 is selected from the group consisting of hydrogen, nitro, amino, isothiocyanato, semicarbazido, thiosemicarbazido, carboxyl, bromoacetamido and maleimido; each R 5 independently is hydrogen or C ⁇ -C 4 alkyl; with the proviso that R 2 and R 4 cannot both be hydrogen but one of R 2 and R 4 must be hydrogen; or a pharmaceutically acceptable salt thereof.
  • Additional preferred functionalized chelant compounds of formula I include those compounds where at least one Q is hydrogen and are represented by formula III:
  • each Q independently is hydrogen or CHR 5 COOR
  • Q 1 is hydrogen or (CHR 5 ) w C0 2 R; with the proviso that at least two the sum of Q and Q 1 must be other than hydrogen and one Q is hydrogen; each R independently is hydrogen benzyl or C1-C 4 alkyl; m is integer from 0 to 5 inclusive; w is 0 or 1;
  • R 2 is selected from the group consisting of hydrogen, nitro, amino, isothiocyanato, semicarbazido, thiosemicarbazido, carboxyl, bromoacetamido and maleimido;
  • R 3 is selected from the group consisting of C 1 -C 4 alkoxy, -OCH 2 COOH, hydroxy and hydrogen;
  • R 4 is selected from the group consisting of hydrogen, nitro, amino, isothiocyanato, semicarbazido, thiosemicarbazido, carboxyl, bromoacetamido and maleimido; each R 5 independently is hydrogen or C1-C 4 alkyl; with the proviso that R 2 and R 4 cannot both be hydrogen but one of R 2 and R 4 must be hydrogen; a pharmaceutically acceptable salt thereof.
  • each Q independently is hydrogen or CHR s 5°Cr00R; with the proviso that at least one Q must be other than hydrogen; each R independently is hydrogen benzyl or C 1 -C 4 alkyl; m is integer from 0 to 5 inclusive;
  • R 2 is selected from the group consisting of hydrogen, nitro, amino, isothiocyanato, semicarbazido, thiosemicarbazido, carboxyl, bromoacetamido and maleimido;
  • R 3 is selected from the group consisting of C 1 -C 4 alkoxy, -OCH 2 COOH, hydroxy and hydrogen;
  • R 4 is selected from the group consisting of hydrogen, nitro, amino, isothiocyanato, semicarbazido, thiosemicarbazido, carboxyl, bromoacetamido and maleimido;
  • each R 5 independently is hydrogen or C 3. -C 4 alkyl; with the proviso that R 2 and R 4 cannot both be hydrogen but one of R 2 and R 4 must be hydrogen; or a pharmaceutically acceptable salt thereof,
  • the functionalized chelants of formula I useful in the practice of the present invention can be prepared by known methods .
  • General synthetic approach to a twelve- membered macrocyclic, bifunctional chelant of the present invention as represented by formula I involves monofuctionalization of a free-base macrocycle (for example, 1, 4, 7, 10-tetraazacyclododecane) at only one of the nitrogen atoms with an appropriate electrophile (for example, any appropriately substituted alpha- halocarboxylic acid ester) .
  • This electrophile must possess a suitable linker moiety which would allow covalent attachment of bifunctional ligand to a biological molecule.
  • Various synthetic routes to functionalized chelants of formula I have been described U.S. Patent Nos. 5,435,990 and 5,652,361, both incorporated herein by reference.
  • 225 Ac radionuclide is not critical to the present invention.
  • 225 Ac can be prepared in a cyclotron.
  • 225 Ac can be obtained in pure form from Department of Energy (DOE), U.S.A., and
  • the 225 Ac conjugates of the present invention can be prepared by first forming the complex and then binding the biological molecule.
  • the process involves preparing or obtaining the ligand, forming the complex with 225 Ac and then adding the biological molecule.
  • the process may involve first conjugation of the ligand to the biological molecule and then the formation of the complex with 225 Ac . Any suitable process that results in the formation of the 225 Ac conjugates of this invention is within the scope of the present invention.
  • BFC bifunctional chelant
  • DOTA 1,4,7,10 tetraazacyclododecane-1,4,7,10- tetraacetic acid
  • MeO-DOTA-NCS 1- [ (2-methoxy ⁇ 5-isothiocyanato- phenyl) -carboxymethyl] -4,7, 10-triscarboxy- methyl-1, 4,7, 10-tetraazacyclododecane;
  • TMAA tetramethyl ammonium acetate buffer
  • Sephadex C-25 resin is a cation exchange resin, sold by Pharmacia Inc.
  • EDTA ethylenediaminetetraacetic acid
  • DTPA diethylenetriaminepentaacetic acid
  • TETA 1, 4, 8, 11-tetraazacyclotetradecane-l, 4, 8, 11- tetraacetic acid
  • DOTPA 1, 4,7, 10-tetraazacyclododecane-l, 4,7,10- tetrapropionic acid
  • TETPA 1, 4, 8, 11-tetraazacyclotetradecane-l, 4, 8, 11- tetrapropionic acid
  • DOTMP 1, 4, 6, 10-tetraazacyclodecane-l, 4, 7, 10- tetramethylenephosphonic acid.
  • the preparation of 225 Ac conjugates involved two steps. First, the 225 Ac complex was prepared by mixing a solution of the functionalized chelant compound of formula I with the solution of 225 Ac at pH of about 5-6 in a suitable buffer. The complex formation was tested using cation exchange chromatography. Then, the conjugation of the 225 Ac complex to a biological molecule (suitably an antibody) was carried out at the pH of about 8.5 in the presence of a suitable buffer. The antibody and the antibody 225 Ac complex conjugates were then separated from the unconjugated low molecular weight materials using gel filtration chromatography. The fraction of radioactivity associated with the antibody was then determined.
  • a biological molecule suitably an antibody
  • the ⁇ emission counting was performed using a 3-inch x 3-inch Nal well crystal utilizing the ⁇ emission of 225 Ac decay product 221 Fr (half-life of 4.8 min.) at 218 KeV. Counting was carried out half an hour after sample preparation.
  • MeO-DOTA-NCS 35 ⁇ l; 0.31 mg/ml
  • 225 Ac chloride solution 35 ⁇ l; 1.65 ⁇ Ci/ ⁇ l,
  • the pH was adjusted to about 5 using the TMAA buffer (130 ⁇ l , 0.2 M, pH about 6). Reaction mixture was incubated at about 50°C for one hour. Complex formation was checked by cation exchange chromatography employing the Sephadex C-25 resin and it was determined that 99 percent of 225 Ac was complexed.
  • HuMl95 (or B4) monoclonal antibody solution was mixed with 0.05 M HEPES containing EDTA at about pH 8 and dialysed against 0.05 M HEPES buffer for 24 hours in an Amicon stirred cell dialysis unit to remove any metal ions associated with antibodies.
  • the immunoconjugate was recovered from the stirred cell and characterized using a size exclusion high pressure liquid chromatography (HPLC) . This was compared to the native HuMl95 or B4.
  • Antibody concentration was determined by UV-absorption at 280 nM.
  • the immunoconjugates could be labeled readily with 1:L1 In showing success of the conjugation reaction.
  • 1:L1 In For example, approximately 400 ⁇ Ci of m In in 200 ⁇ l of 0.2 M HC1 was mixed with 29 ⁇ l of ammonium acetate (3M) and 9 ⁇ l of J-ascorbic acid (150 mg/ml) to adjust the pH to 5.0 and then 0.5 mg of HuM195-MeO-DOTA-NCS immunoconjugate was added. The reaction was allowed to progress at 37°C for 60 minutes. An 87% reaction yield was obtained.
  • Example 4 Labeling of HuMl95-MeO-DOTA
  • the three 225 Ac-HuM195-MeO-DOTA solutions from Example 4 were combined and challenged with 20 ⁇ l of 10 mM DTPA to remove unbounded metals. 100 ⁇ l of 1-ascorbic acid (150 mg/ml) was added as a radioprotection agent. The solution was purified through a 10-DG desalting column (Bio-Rad company) to separate 225 Ac-HuMl95-MeO-DOTA from unreacted
  • the purified 225 Ac-HuM195-MeO-DOTA was assessed for stability in different media such as 1% and 25% albumin (human) and 1% and 25% human serum at 37°C. Overall stability half-lives of 225 Ac-HuMl95-MeO-DOTA in either albumin (human) at 4 °C or human serum at 37 °C exceeded 150 days.

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Abstract

Cette invention a trait à des complexes d'actinium-225 (225Ac) possédant des chelatants fonctionnalisés correspondant à la formule (I). Dans cette formule, chaque Q représente, de manière indépendante, un hydrogène ou (CHR5)pCO2R, Q1 représente un hydrogène ou (CHR5)wCO2R, chaque R représente, de manière indépendante, un hydrogène, un benzyle ou un alkyle porteur de 1 à 4 atomes de carbone, à la condition qu'au moins deux éléments formés par la somme de Q et de Q1 représentent un autre atome que celui de l'hydrogène, chaque R5 représente, de manière indépendante un hydrogène, un alkyle porteur de 1 à 4 atomes de carbone ou un (C¿1?-C2 alkyl)phényle, X et Y représentent, chacun de manière indépendante, un hydrogène ou peuvent, pris avec un X et un Y adjacents, former une liaison supplémentaire carbone-carbone, la valeur de n est 0 ou 1, m représente un nombre entier dont la valeur est comprise entre 0 et 10 inclus, la valeur de p est 1 ou 2, celle de r est 0 ou 1, celle de w est 0 ou 1, à la condition que la valeur de n ne soit que 1 lorsque X et/ou Y constituent une liaison supplémentaire carbone-carbone et que la somme de r et de w s'échelonne entre 0 et 1, L représente un groupe lieur/séparateur lié de façon covalente à un atome d'hydrogène de l'un des atomes de carbone auquel il est relié et remplace celui-ci, ce groupe lieur/séparateur correspondant à la formule (1), dans laquelle S représente un nombre entier d'une valeur s'échelonnant de 0 à 1, t représente un nombre entier d'une valeur s'échelonnant de 0 à 20 inclus, R?1¿ représente un fragment électrophile ou nucléophile permettant un rattachement covalent à un anticorps ou à un fragment de celui-ci ou bien un lieur de synthèse pouvant être rattaché à un anticorps ou à un fragment ou à un précurseur de celui-ci, Cyc représente un fragment aliphatique cyclique, un fragment aromatique, un fragment aliphatique hétérocyclique ou un fragment aromatique hétérocyclique, chacun desdits fragments étant éventuellement substitués par un ou plusieurs groupes n'interférant pas avec la liaison à un anticorps ou à un fragment d'anticorps, à la condition que, lorsque la valeur de s, t, m, r, et de n est 0, R1 représente autre chose qu'un carboxyle. Cette invention, qui porte également sur les sels acceptables du point de vue pharmaceutique, ainsi que sur les conjugats de ces complexes, concerne, de surcroît, l'utilisation qui en est faite en matière de radio-immunothérapie.
PCT/US2001/005927 2000-02-25 2001-02-23 Complexes et conjugat d'actinium-225 pour radio-immunotherapie WO2001066155A2 (fr)

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HU0203743A HUP0203743A2 (en) 2000-02-25 2001-02-23 Actinium-225 complexes and conjugates for radioimmunotherapy
AU49069/01A AU4906901A (en) 2000-02-25 2001-02-23 Actinium-225 complexes and conjugates for radioimmunotherapy
EP01922246A EP1227849A2 (fr) 2000-02-25 2001-02-23 Complexes et conjugat d'actinium-225 pour radio-immunotherapie
CA2381123A CA2381123C (fr) 2000-02-25 2001-02-23 Complexes et conjugat d'actinium-225 pour radio-immunotherapie

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US60/185,220 2000-02-25

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WO2002005859A2 (fr) * 2000-07-04 2002-01-24 Anticancer Therapeutic Inventions As Procede de radiotherapie
WO2002022000A3 (fr) * 2000-09-15 2002-05-16 Sloan Kettering Inst Cancer Therapie par particules alpha ciblees utilisant des conjugues d'actinium-225
WO2002067999A2 (fr) * 2001-02-28 2002-09-06 Dow Global Technologies Inc. Actinium-225 complexes and conjugates for targeted radiotherapy
WO2004091668A1 (fr) * 2003-04-15 2004-10-28 Algeta As Thorium 227 utilisable en radiotherapie pour traiter une maladie des tissus mous
JP2006523664A (ja) * 2003-04-15 2006-10-19 アルゲッタ エイエス 軟組織疾患の放射線治療におけるトリウム−２２７の使用
US9724436B2 (en) 2010-02-12 2017-08-08 Bayer As Alpha-emitting complexes
EP3142710A4 (fr) * 2014-05-16 2018-04-25 Memorial Sloan Kettering Cancer Center Marquage en une étape par actinium-225 d'anticorps pour obtenir une haute activité spécifique
CN111808161A (zh) * 2020-06-01 2020-10-23 北京大学 一种对生物化合物进行放射性标记的方法
WO2022006269A1 (fr) * 2020-06-30 2022-01-06 The Johns Hopkins University Combinaison non intuitive de supports d'administration médicamenteuse du même médicament pour retarder la croissance synergique de tumeurs solides
EP3886921A4 (fr) * 2018-11-30 2022-11-23 Actinium Pharmaceuticals, Inc. Anticorps conjugués à l'actinium 225 et l'actinium 227, compositions et procédés associés

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Cited By (21)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002005859A2 (fr) * 2000-07-04 2002-01-24 Anticancer Therapeutic Inventions As Procede de radiotherapie
WO2002005859A3 (fr) * 2000-07-04 2002-09-06 Anticancer Therapeutic Inv Sa Procede de radiotherapie
US7335154B2 (en) 2000-07-04 2008-02-26 Algeta As Radiotherapy
US7056275B2 (en) 2000-07-04 2006-06-06 Algeta A/S Radiotherapy
WO2002022000A3 (fr) * 2000-09-15 2002-05-16 Sloan Kettering Inst Cancer Therapie par particules alpha ciblees utilisant des conjugues d'actinium-225
EP1317268A2 (fr) * 2000-09-15 2003-06-11 Sloan Kettering Institute For Cancer Research Therapie par particules alpha ciblees utilisant des conjugues d'actinium-225
EP1317268A4 (fr) * 2000-09-15 2004-12-15 Sloan Kettering Institutefor C Therapie par particules alpha ciblees utilisant des conjugues d'actinium-225
WO2002067999A2 (fr) * 2001-02-28 2002-09-06 Dow Global Technologies Inc. Actinium-225 complexes and conjugates for targeted radiotherapy
WO2002067999A3 (fr) * 2001-02-28 2002-10-24 Dow Global Technologies Inc Actinium-225 complexes and conjugates for targeted radiotherapy
US6670456B2 (en) 2001-02-28 2003-12-30 Dow Global Technologies Inc. Actinium-225 complexes and conjugates for targeted radiotherapy
JP2006523664A (ja) * 2003-04-15 2006-10-19 アルゲッタ エイエス 軟組織疾患の放射線治療におけるトリウム−２２７の使用
EA008195B1 (ru) * 2003-04-15 2007-04-27 Алгета Ас Применение тория-227 в лучевой терапии заболеваний мягких тканей
WO2004091668A1 (fr) * 2003-04-15 2004-10-28 Algeta As Thorium 227 utilisable en radiotherapie pour traiter une maladie des tissus mous
JP2012051942A (ja) * 2003-04-15 2012-03-15 Algeta As 軟組織疾患の放射線治療におけるトリウム−２２７を用いた医薬組成物、複合体及びその調製方法、並びにキット
US9724436B2 (en) 2010-02-12 2017-08-08 Bayer As Alpha-emitting complexes
US10682430B2 (en) 2010-02-12 2020-06-16 Bayer As Alpha-emitting complexes
EP3142710A4 (fr) * 2014-05-16 2018-04-25 Memorial Sloan Kettering Cancer Center Marquage en une étape par actinium-225 d'anticorps pour obtenir une haute activité spécifique
AU2015258801B2 (en) * 2014-05-16 2020-10-08 Memorial Sloan Kettering Cancer Center One-step labeling of antibodies to high specific activity with actinium-225
EP3886921A4 (fr) * 2018-11-30 2022-11-23 Actinium Pharmaceuticals, Inc. Anticorps conjugués à l'actinium 225 et l'actinium 227, compositions et procédés associés
CN111808161A (zh) * 2020-06-01 2020-10-23 北京大学 一种对生物化合物进行放射性标记的方法
WO2022006269A1 (fr) * 2020-06-30 2022-01-06 The Johns Hopkins University Combinaison non intuitive de supports d'administration médicamenteuse du même médicament pour retarder la croissance synergique de tumeurs solides

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ZA200200659B (en) 2003-01-23
CA2381123A1 (fr) 2001-09-13
WO2001066155A8 (fr) 2002-02-07
EP1227849A2 (fr) 2002-08-07
AU4906901A (en) 2001-09-17
WO2001066155A3 (fr) 2002-06-06
HUP0203743A2 (en) 2003-02-28
CA2381123C (fr) 2011-10-04

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