[go: up one dir, main page]

WO2000059896A1 - 225ac-heha et composes, procedes de synthese et procedes d'utilisation correspondants - Google Patents

225ac-heha et composes, procedes de synthese et procedes d'utilisation correspondants Download PDF

Info

Publication number
WO2000059896A1
WO2000059896A1 PCT/US2000/007643 US0007643W WO0059896A1 WO 2000059896 A1 WO2000059896 A1 WO 2000059896A1 US 0007643 W US0007643 W US 0007643W WO 0059896 A1 WO0059896 A1 WO 0059896A1
Authority
WO
WIPO (PCT)
Prior art keywords
heha
bifunctional
targeting agent
compound
group
Prior art date
Application number
PCT/US2000/007643
Other languages
English (en)
Inventor
Martin W. Brechbiel
Kim Deal
Original Assignee
The United States Of America, Represented By The Secretary, Department Of Health And Human Services
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 The United States Of America, Represented By The Secretary, Department Of Health And Human Services filed Critical The United States Of America, Represented By The Secretary, Department Of Health And Human Services
Priority to AU44501/00A priority Critical patent/AU4450100A/en
Priority to US09/937,030 priority patent/US6696551B1/en
Publication of WO2000059896A1 publication Critical patent/WO2000059896A1/fr
Priority to US10/767,133 priority patent/US6995247B2/en

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B59/00—Introduction of isotopes of elements into organic compounds ; Labelled organic compounds per se
    • C07B59/002—Heterocyclic compounds
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D259/00—Heterocyclic compounds containing rings having more than four nitrogen atoms as the only ring hetero atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
    • C07F9/02—Phosphorus compounds
    • C07F9/547—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom
    • C07F9/6524—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having four or more nitrogen atoms as the only ring hetero atoms

Definitions

  • the present invention relates to a chelation complex comprising 225 Actinium ( 225 Ac) and 1,4,7,10,13,16- hexaazacyclohexadecane-N,N' , N", N'", N "", N' ' ' ' '-hexaacetic acid (HEHA) ( 225 Ac-HEHA) , bifunctional HEHA, bifunctional HEHA-targeting agent, bifunctional 225 Ac-HEHA, bifunctional 225 Ac-HEHA-targeting agent, and methods of synthesis and use, such as in the context of radioimmunotherapy, decontamination and detoxification.
  • HEHA 1,4,7,10,13,16- hexaazacyclohexadecane-N,N' , N", N'", N "", N' ' ' ' ' '-hexaacetic acid
  • HEHA 1,4,7,10,13,16- hexaazacyclohexadecane-N,N
  • the radioisotope chosen is determined, at least in part, by the type of disease to be treated. The reason for this is that the type of particles emitted by a given radioisotope are directly related to tissue penetration and the ability of the isotope to kill cells (Boll et al . , Radiochim. Acta 79: 87-91 (1997)). ⁇ -emitters, like 90 Y and 131 I, which have a tissue range of several millimeters, have been used successfully to treat solid tumors (Boll et al . (1997) , supra) .
  • tissue range of several millimeters is not optimal for the treatment of single cells, small clusters of cells, micrometastatic disease, leukemias and lymphomas (Jurcic et al . , In: Cancer Chemotherapy and Biological Response Modifiers Annual 27, Pinedo et al . , eds . , New York: Elsevier B.V. (1998), pp. 195-216; Falini et al . , Cancer Surveys 30: 295-309
  • ⁇ -emitters combine high cytotoxicity with a short tissue range, i.e., less than about 150 ⁇ (Boll et al . (1997), supra) .
  • 225 Ac decays through a chain of four ⁇ emissions and two ⁇ emissions to the stable isotope 209 Bi, thereby releasing a large amount of energy (28 MeV) (Davis et al . , Nucl . Med. Biol . , accepted; Alleluia et al . , In: Gmelin Handbook of Inorganic Chemistry, 8 th ed. , Kugler et al., eds., New York: Springer-Verlag (1981), pp. 181- 193) . Unfortunately, most of the 225 Ac administered in a dose is deposited in the liver and bone (Beyer et al . , Isotopenpraxis 26: 111-114 (1990)). Thus, numerous attempts have been made to reduce the toxicity of 225 Ac through chelation with, for example, citrate (Beyer et al. (1990), supra), EDTMP
  • the present invention provides an ⁇ -particle- emitting radioisotope chelation complex comprising 225 Actinium ( 225 Ac) and 1,4,7,10,13,16- hexaazacyclohexadecane- N,N' , N", N'", N"", N'""-hexaacetic acid (HEHA) ( 225 Ac-HEHA) .
  • HEHA 1,4,7,10,13,16- hexaazacyclohexadecane- N,N' , N", N'", N"", N'”"-hexaacetic acid
  • a bifunctional HEHA which can chelate a radiosotope, in particular 25 Ac, and can be attached to a targeting agent, such as a bifunctional HEHA having one of the following formulae:
  • R is C0 2 H, CONHR' , P(0)R'OH or P(O) (OR 1 ) OH
  • R' is H, a Ci-Cg alkyl, phenyl or benzyl, wherein said phenyl or benzyl is unsubstituted or substituted
  • n is 1-6
  • X is N0 2 , NH 2 , NCS, NHC(0)CH 2 Z (in which Z is Cl, Br or I), or
  • bifunctional HEHAs are set forth herein.
  • a compound comprising the bifunctional HEHA conjugated to a targeting agent is also provided.
  • a bifunctional 225 Ac-HEHA complex comprising 225 Ac complexed with the bifunctional HEHA described above as well as a compound comprising the bifunctional 25 Ac- HEHA complex conjugated to a targeting agent.
  • the present invention further provides a method of making HEHA.
  • the method comprises preparing the free base of 1,4,7,10,13,16- hexaazacyclohexadecane under anhydrous conditions, azeotropically removing trace water with benzene, N- alkylating the macrocycle to produce the hexaester, saponifying the hexaester, and purifying HEHA.
  • the hexaester is produced by reacting the free base with Na 2 C0 3 and tert-butyl bromoacetate in anhydrous CH 3 CN.
  • a method of making a bifunctional HEHA comprises the preparation of a tert-butyloxycarbonyl protected iminodiacetic acid that is condensed with an amino acid ester.
  • the resulting diester is then saponified with base, and after acidification, converted to a disuccinimidyl ester.
  • This active diester is then reacted with an N-2-aminoethyl amide of para-nitrophenylalanine that introduces the latent bifunctionality aspect that will be unmasked.
  • the protecting group is removed by treatment with acid, and the amide carbonyl functional groups are reduced via diborane.
  • the resulting macrocyclic polyamine is isolated as the protonated salt.
  • the free base is generated and then the free amines are alkylated to introduce protected R groups.
  • the protected R groups are then deprotected.
  • the nitro group is then hydrogenated to the aniline, which is then converted to an isothiocyanate, a haloacetamide or a maleimide for conjugation to a targeting agent.
  • the method can further comprise the conjugation of a bifunctional HEHA to a targeting agent.
  • the method comprises the preparation of a cyclic hexapeptide that comprises para- nitrophenylalanine or ⁇ -protected lysine and the subsequent reduction of amide carbonyl functional groups.
  • the resulting macrocyclic polyamine is isolated as the protonated salt .
  • the free base is then generated and the free amines are alkylated to introduce protected R groups, which are subsequently deprotected.
  • the nitro group is hydrogenated to the aniline and the aniline is converted to an isothiocyanate, a haloacetamide or a maleimide, any one of which can then be conjugated to a targeting agent.
  • a method of treating disease comprises administering to a patient having disease a disease-treatment effective amount of a 25 Ac-HEHA targeting agent as described above in which the targeting agent is specific for diseased cells.
  • a method of treating cancer comprises administering to a patient having cancer a cancer- treatment effective amount of a 22S Ac-HEHA-targeting agent as described above in which the targeting agent is specific for the cancer to be treated.
  • a method of treating a solid tumor is provided. The method comprises intratumorally administering to a patient having a tumor a tumor- treatment effective amount of 225 Ac-HEHA or 25 Ac-HEHA- targeting agent in which the targeting agent is specific for the tumor.
  • the method further comprises simultaneously or sequentially peritumorally administering to the patient HEHA in an amount effective to chelate any radioactive decay products from the compound.
  • a method of decontaminating a sample from 225 Ac is provided. The method comprises contacting the sample with a decontaminating-effective amount of HEHA.
  • a further embodiment is a method of decontaminating a person who has been externally contaminated with 22S Ac .
  • the method comprises contacting the person with a decontaminating-effective amount of HEHA.
  • a method of detoxifying a person who has internalized 225 Ac is provided.
  • the method comprises administering to the person a detoxifying-effective amount of HEHA.
  • the present invention is predicated on the surprising and unexpected discovery that HEHA chelates 225 Ac in such a manner as to provide sufficient in vivo stability to enable its use in the context of radioimmunotherapy and other contexts. Accordingly, in one embodiment, the present invention provides an ⁇ - particle-emitting radioisotope chelation complex comprising 225 Ac and HEHA.
  • 25 Ac-HEHA is highly desirable because it comprises 225 Ac, which is a metal radioisotope with excellent cytotoxicity that forms a suitable complex with HEHA having a half-life (i.e., approximately 10.5 days, wherein from about 30 min to about 3 wks is preferred and from about 30 min to about 11 days is more preferred) and an emission quality that are characteristic of radiopharmaceuticals and a toxic radioactive decay chain that results in nonradioactive material as a final product.
  • a half-life i.e., approximately 10.5 days, wherein from about 30 min to about 3 wks is preferred and from about 30 min to about 11 days is more preferred
  • an emission quality that are characteristic of radiopharmaceuticals and a toxic radioactive decay chain that results in nonradioactive material as a final product.
  • the present invention further provides a bifunctional HEHA which can chelate a radiosotope, which is preferably 225 Ac, and can attach to a targeting agent, such as described herein.
  • HEHA can be rendered bifunctional in any suitable manner in accordance with methods known in the art (see, e.g., Wang, Chemistry of Protein Conjugation and Crosslinking, CRC Press, Boca Raton, Florida (1991) ; Lundblad, Chemical Reagents for Protein Modification. CRC Press, Boca Raton, Florida (1991) ) .
  • Preferred bifunctional HEHAs include those of the following formulae:
  • R is selected from the group consisting of C0 2 H, CONHR', P(0)R'0H and P(0) (OR 1 )0H
  • R' is selected from the group consisting of
  • n 1-6 and, X is selected from the group consisting of N0 2 , NH 2 , NCS, NHC(0)CH 2 Z (in which Z is selected from the group consisting of Cl, Br and I) , and
  • R is C0 2 H and R' is H or CH 3
  • R' is H or CH 3
  • R * is phenyl or benzyl, phenyl or benzyl can be substituted with one or more substituents selected from the group consisting of a C.-C 6 alkyl, a halogen, a alkoxy, a C--C 6 hydroxyl, and a poly-hydroxyl .
  • bifunctional HEHAs include those of formulae:
  • R, R' , n and x are as defined above and R 3 is selected from the group consisting of H, a C--C 6 alkyl, and benzyl .
  • the present invention provides a compound comprising the above-described bifunctional HEHA conjugated to a targeting agent.
  • targeting agent is meant any means that enables specific interaction with a target.
  • the targeting agent can bind to a defined population of cells, for example, through a receptor, a substrate, an antigenic determinant or another binding site on the target cell population.
  • Cell-surface molecules that are cancer specific antigens (or disease-specific antigens) and can serve as targets are known in the art.
  • cancer-specific, cell-surface molecules include placental alkaline phosphatase (testicular and ovarian cancer) , pan carcinoma (small cell lung cancer) , polymorphic epithelial mucin (ovarian cancer) , prostate- specific membrane antigen, ⁇ -fetoprotein, B-lymphocyte surface antigen (B-cell lymphoma) , truncated EGFR (gliomas) , idiotypes (B-cell lymphoma) , gp95/gp97 (melanoma) , N-CAM (small cell lung carcinoma) , cluster w4 (small cell lung carcinoma) , cluster 5A (small cell carcinoma) , cluster 6 (small cell lung carcinoma) , PLAP (seminomas, ovarian cancer, and non-small cell lung cancer) , CA-125 (lung and ovarian cancers) , ESA (carcinoma), CD19, 22 or 37 (B-cell lymphoma), 250 kD proteoglycan (
  • cancer-specific, cell-surface receptors examples include erbB-2, erbB-3, erbB-4, IL-2 (lymphoma and leukemia) , IL-4 (lymphoma and leukemia) , IL-6 (lymphoma and leukemia) , MSH (melanoma) , transferrin (gliomas) , tumor vasculature integrins, and the like.
  • Preferred cancer-specific, cell-surface receptors include erbB-2 and tumor vasculature integrins, such as CDlla, CDllb, CDllc, CD18, CD29, CD51, CD61, CD66d, CD66e, CD106, and CDW145 .
  • the erbB-2 receptor has been found in breast, ovarian, gastric, salivary gland and adeno-carcinomas and in non-small cell carcinomas of the lung. Over-expression of the erbB-2 receptor on such cancers has been found to correlate with poor prognosis. In vi tro studies strongly suggest that over-expression of erbB-2 may play an important role in tumor progression.
  • a single-chain antibody scAb is that which binds c-erbB-2 (WO 93/16185) . See, also, WO 93/21232 and http: //www. antibody resource.com for antibody sequences that can be used to construct scAbs .
  • ScAbs can be developed, based on such antibodies, using techniques known in the art (see, for example, Bind et al . , Science 242: 423-426 (1988), and Whitlow et al . , Methods 2(2): 97-105 (1991)).
  • binding domains include the EGF domain of ⁇ -heregulin, ⁇ -integrin domain, tumor vasculature peptide motifs, and those described in the htt : //ampere .doe-mbi .ucla.edu: 880l/dat/dip.dat and http : //bones .biochem. ualberta . ca/pedro/rt-1.html . databases.
  • Alpha-heregulin is a ligand with affinity for breast cancer cells expressing the human epidermal growth factor receptors erbB-2, erbB-3 and erbB-4. Heregulin interacts indirectly with erbB-2 via heterodimerization with erbB-3 or erbB-4.
  • a targeting agent examples include an "immunological agent,” which is used herein to refer to an antibody, such as a polyclonal antibody or a monoclonal antibody, an immunologically reactive fragment of an antibody, an engineered immunoprotein and the like, a protein (target is receptor, as substrate, or regulatory site on DNA or RNA) , a peptide (target is receptor) , a nucleic acid (target is complementary nucleic acid) , a steroid (target is steroid receptor) , and the like.
  • an antibody such as a polyclonal antibody or a monoclonal antibody, an immunologically reactive fragment of an antibody, an engineered immunoprotein and the like
  • a protein target is receptor, as substrate, or regulatory site on DNA or RNA
  • a peptide target is receptor
  • nucleic acid target is complementary nucleic acid
  • steroid target is steroid receptor
  • Preferred targeting agents include an antibody or an iummunologically reactive fragment thereof, a peptide, e.g., bombesin, gastrin-releasing peptide, RGD peptide, substance P, neuromedin-B, neuromedin-C, somatostatin, octreotide analogues, and metenkephalin, and a hormone, e.g., estradiol, neurotensin, melanocyte stimulating hormone, follicle analogues stimulating hormone, leutenizing hormone, and human growth hormone.
  • Other suitable targeting agents include serum proteins, fibrinolytic enzymes, and biological response modifiers, such as interleukin, interferon, erythropoietin, and colony-stimulating factor.
  • Analogs of targeting agents that retain the ability to bind to a defined target also can be used.
  • synthetic targeting agents can be designed, such as to fit a particular epitope .
  • the targeting agent can include any linking group that can be used to join a targeting agent to, in the context of the present invention, a chelate. It will be evident to one skilled in the art that a variety of linking groups, including bifunctional reagents, can be used.
  • the present invention further provides a bifunctional 225 Ac-HEHA comprising 225 Ac complexed with a bifunctional HEHA as described above. Also, in this regard, the present invention provides a compound comprising a bifunctional 225 Ac-HEHA conjugated to a targeting agent as described above.
  • HEHA HEHA-protein compound
  • a method of making HEHA comprises preparing the free base of the macrocycle 1,4,7,10,13,16- hexaazacyclohexadecane under anhydrous conditions, azeotropically removing trace water with benzene, N- alkylating the macrocycle to produce the hexaester, saponifying the hexaester, and purifying HEHA.
  • the hexaester is produced by reacting the free base with Na 2 C0 3 and tert-butyl bromoacetate in anhydrous CH 3 CN.
  • a preferred method is set forth in Example 1.
  • 225 Ac can be chelated either before or after the chelator is conjugated to a targeting agent.
  • the order chosen can take into account stability and other factors and is well within the ordinary skill in the art.
  • Methods of complexing metal ions with chelants are known and are within the level of ordinary skill in the art .
  • a metal can be incorporated into a chelant moiety by one of three general methods, i.e., direct incorporation, template synthesis and/or transmetallation. Direct incorporation is preferred. Generally, the metal is titrated from substoichiometric levels up to full incorporation, thus eliminating the need for dialysis and extensive chromatographic purification. In this manner, significant losses as well as dilution are avoided.
  • a water-soluble form of the metal such as an inorganic salt
  • a water-soluble form of the metal is dissolved in an appropriate volume of distilled, deionized water, preferably in a dilute acid medium having a pH of from about 1 to about 7 and most preferably at a pH of from about 4 to about 6.
  • Ambient temperatures of about 20 °C to 27 °C or below (to just above freezing) can be readily employed with stirring for metal chelation.
  • Any appropriate metal salt, either in solid form or in solution can be contacted with the chelate, either free in solution or conjugated to a targeting agent, in order to form the chelated 225 Ac .
  • the pH of the mixture is raised slowly by addition of base, typically 0.1 M NaOH, until the donor groups of the polychelant are deprotonated, generally in the pH range of from about 5 to about 9, depending on the chelant moieties. Particular care must be taken to maintain the pH below 8 to avoid precipitation of the metal hydroxide .
  • Preferred methods include those set forth in Examples 2 and 6.
  • a wide variety of metal salts can be employed including, for example, nitrates, iodides, chlorides, citrates, acetates and the like. The choice of an appropriate metal salt as well as the choice of a particularly appropriate chelate for any given metal is within the ordinary skill in the art.
  • a method of making a bifunctional HEHA is provided.
  • Preferred methods include those set forth in Example 3.
  • the method comprises the preparation of a tert-butyloxycarbonyl protected iminodiacetic acid that is condensed with an amino acid ester.
  • the resulting diester is then saponified with base, and after acidification, converted to a disuccinimidyl ester.
  • This active diester is then reacted with an N-2-aminoethyl amide of para-nitrophenylalanine that introduces the latent bifunctionality aspect that will be unmasked.
  • the protecting group is removed by treatment with acid, and the amide carbonyl functional groups are reduced via diborane .
  • the resulting macrocyclic polyamine is isolated as the protonated salt.
  • the free base is generated and then the free amines are alkylated to introduce protected R groups .
  • the protected R groups are then deprotected.
  • the nitro group is then hydrogenated to the aniline, which is then converted to an isothiocyanate, a haloacetamide or a maleimide for conjugation to a targeting agent.
  • bifunctional HEHA reagents are via the preparation of the cyclic hexapeptide, wherein the amino acid components provide the latent bifunctionality aspect that will be unmasked, specifically an amino acid such as of para- nitrophenylalanine or an ⁇ -protected lysine.
  • the advantage to this method is the potential to introduce additional functional groups into the macrocyclic ring stereospecifically and thus tune the cavity size of the macrocycle.
  • the ring is formed from the linear hexapeptide by cyclization with an activating and/or dehydrating reagent, for example DPPA (diphenylphosphoryl azide) .
  • the carbonyl functional groups are reduced via diborane .
  • the free amines in the resulting macrocyclic polyamine are then alkylated to introduce protected R groups.
  • the protected R groups are then deprotected.
  • the nitro group is then hydrogenated to the aniline, which is then converted to an isothiocyanate, a haloacetamide or a maleimide for conjugation to a targeting agent.
  • Bodanszky (1993), supra Green et al . (1991), supra ; Wong (1991), supra ; Lundblad (1991) , supra ; Magerstadt (1991) , supra ; and Aston et al., Tetrahedron Lett . 35:3687-3690 (1994).
  • 225 Ac-HEHA wherein the HEHA is bifunctional, or the bifunctional HEHA is to be conjugated
  • 225 Ac-HEHA or the bifunctional HEHA is mixed in aqueous solution with the desired targeting agent, such as an antibody, at a pH of from about 6 to about 11, most preferably at a pH of from about 7 to about 9.5.
  • the pH is adjusted with a buffered solution, such as a bicarbonate buffered solution.
  • a buffered solution such as a bicarbonate buffered solution.
  • the temperature of the solution can range from just above freezing to the temperature at which the chelate becomes unstable or the protein denatures. Often temperatures above 37 °C tend to denature proteins.
  • chelate and targeting agent are mixed in a molar ratio of greater than 1:1 and less than 100:1 depending on protein concentration. Ratios of about 2:1 to about 4:1 are preferred, but the choice of reaction conditions is within the ordinary skill in the art.
  • a method of attaching a bifunctional HEHA to a targeting agent comprises the conjugation of a bifunctional HEHA to a targeting agent or vector.
  • Preferred methods include those set forth in Example 4.
  • the targeting agent was a monoclonal antibody.
  • Typical initial ratios used to achieve the desired final chelate/protein ratio of 1-2 were a 10-fold initial excess of ligand for whole antibodies and a 10-15-fold initial excess of ligand for antibody fragments.
  • the reaction mixture was purified by centrifugation filtration.
  • dialysis against 1 1 of 0.15 M NH 4 OAc for a minimum of 6 hr and changing buffer for a total of 4 times provided the conjugate ready for radiolabeling.
  • the conjugates can be used as such with appropriate pH adjustment, if needed.
  • the product can be purified.
  • standard purification techniques known in the art including column chromatography and high performance liquid chromatography (HPLC) .
  • 225 Ac-HEHA and conjugates thereof can be administered in vivo in the form of a composition, e.g., a pharmaceutical composition, comprising a carrier, e.g., pharmaceutically acceptable carrier.
  • a carrier e.g., pharmaceutically acceptable carrier.
  • a biologically acceptable, normal saline solution can be appropriately employed.
  • the carrier can include a minor amount of a carrier protein, such as human serum albumin, for example, to stabilize the targeting agent.
  • Stabilizers, antioxidants, osmolality adjusting agents, buffers, pH adjusting agents, etc. can be included in the composition.
  • the composition can be in the form of a solution, suspension or dispersion.
  • Suitable additives include, for example, physiologically biocompatible buffers, additions of chelants or calcium chelate complexes, or optionally, additions of calcium or sodium salts.
  • Parenterally administrable forms e.g., intravenous forms, should be sterile and free from physiologically unacceptable agents and should have low osmolality to minimize irritation or other adverse effects upon administration.
  • Suitable vehicles include aqueous vehicles customarily used for administering parenteral solutions, such as sodium chloride injection, Ringer's injection, dextrose injection, dextrose and sodium chloride injection. Lactated Ringer's injection and other solutions are as described in Remington' s Pharmaceutical Sciences. 15th ed. , Easton: Mack Publishing Co. (1975) .
  • the solutions can contain preservatives, antimicrobial agents, buffers and antioxidants conventionally used for parenteral solutions, excipients and other additives that are compatible with the chelates and will not interfere with the manufacture, storage or use of products.
  • concentration of 225 Ac-HEHA or conjugate thereof in a composition will be a matter of choice. Levels of 0.5 mg/ml are readily attainable but the concentration may vary considerably depending upon the specifics of any given application. Appropriate concentrations of biologically active materials in a carrier are routinely determined in the art .
  • the effective dose (referred to herein as "disease- treatment effective amount,” “cancer-treatment effective amount,” “tumor-treatment effective amount,” “decontaminating effective amount” and “detoxifying- effective amount”) of HEHA, 25 Ac-HEHA or conjugate of either of the foregoing to be utilized for any application will also depend upon the particulars of that application.
  • the dose will depend, inter alia, upon tumor burden, accessibility, route of administration, administration of other active agents, and the like.
  • a therapeutically effective dose is from about 20 mCi to about 300 mCi .
  • HEHA and 225 Ac-HEHA-targeting agent can be administered in accordance with the present inventive methods by any suitable route .
  • routes include intravenous, intraperitoneal, and the like, depending on the site of contamination with 224 Ac or the disease or cancer to be treated, respectively, the location of the contaminated/diseased/cancerous cells, the extent of contamination/disease/cancer, and other factors.
  • the determination of the appropriate route (s) of administration for a given application is within the ordinary skill in the art.
  • transurethral delivery to the prostate or periprostate space or transrectal injection can be used.
  • the conjugates of the present invention are introduced into the body and are allowed to concentrate in the target region.
  • the therapeutic effect occurs when the conjugates are near or in contact with and bind to the targeted cells.
  • Cell death can be a direct or indirect result of the radiation event of the radiometal that is positioned in close proximity to the cell.
  • the conjugate comprises a monoclonal antibody that is specific (e.g., for a cell-surface molecule) for a cell, such as a diseased cell, to be killed.
  • Cell death is caused by decay of the radiometal and can occur in one of two ways .
  • a single hit in the cell nucleus can be cytotoxic.
  • the isotope to which the radiometal decays after emitting the alpha particle is ejected from the chelate on a trajectory opposite that of the alpha particle.
  • the bound cell therefore, can still be hit even when the alpha particle is emitted on a trajectory away from the cell .
  • a single hit in the cell membrane by the decayed isotope can cause irreparable cell injury leading to cell death.
  • the relatively high effectiveness of the alpha particle means that less radioactive material can be employed. Selectivity of the targeting agent, e.g., monoclonal antibody, and the short range (a few cell diameters) of the alpha particles minimizes the destruction of healthy tissue on a cellular level.
  • the benefits that inure to this embodiment of the invention are numerous.
  • the high specificity of the conjugate minimizes total radiation dosage. Only enough radiation for the target cells need be employed.
  • 22S Ac-HEHA is cleared rapidly from the body should the targeting agent be disrupted.
  • the amount of radiometal employed is minimized, the radiation hazard to persons preparing and administering the conjugate is significantly reduced.
  • tissue damage or whole body dose during therapy is markedly reduced as compared to that from presently employed methods of radiation therapy, such as isotope implants, external radiation therapy, and immunoradiotherapy employing iodine-131 labeled polyclonal or autologous antibodies.
  • both biological and physical half-lives of the targeting radiobiological can now be controlled, minimizing whole body radiation effects.
  • a therapeutic dose is delivered specifically to malignant cells, either localized or metastasized.
  • the ability of conjugates to provide an effective dose of therapeutic radiation specifically to metastasized cells is also unique and singularly useful for cancer therapy.
  • 225 Ac-HEHA or 225 Ac-HEHA-targeting agent is administered intratumorally . Any leakage of decay products to the surrounding healthy tissue can be chelated by the subsequent or simultaneous administration of HEHA or the like.
  • a 225 Ac- HEHA-targeting agent that targets leukemic cells or prostate cancer cells is administered in the treatment of leukemia or prostate cancer, respectively.
  • the 225 Ac- HEHA-targeting agent desirably is internalized by the cell and all decay occurs within the cell, preferably with the daughter isotopes decaying thereafter within one hour or less, thereby minimizing any damage to normal cells and tissue.
  • the present invention provides a method of treating disease.
  • the method comprises administering to a patient having disease a disease- treatment effective amount of 225 Ac-HEHA-targeting agent in which the targeting agent is specific for diseased cells.
  • the targeting agent is an antibody.
  • a method of treating cancer The method comprises administering to a patient having cancer a cancer-treatment effective amount of 225 Ac-HEHA- targeting agent in which the targeting agent is specific for the cancer.
  • the targeting agent is an antibody.
  • the method preferably comprises intratumorally administering to a patient having a tumor a tumor-treatment effective amount of 225 Ac-HEHA or 225 Ac-HEHA-targeting agent, in which the targeting agent is specific for the tumor. Any leakage of decay products to the surrounding healthy tissue can be chelated by the subsequent or simultaneous administration of HEHA or the like.
  • a method of decontaminating a sample from 225 Ac comprises contacting the sample with a decontaminating-effective amount of HEHA.
  • a decontaminating-effective amount can be determined in accordance with methods known in the art.
  • the HEHA is attached to a solid support and the sample is a liquid. This method has application in the context of bioremediation .
  • a method of decontaminating a person who has been externally contaminated with 225 Ac comprises contacting the person with a decontaminating-effective amount of HEHA.
  • a decontaminating-effective amount can be determined in accordance with methods known in the art.
  • a method of detoxifying a person who has internalized 225 Ac is also provided.
  • the method comprises administering to the person a detoxifying-effective amount of HEHA.
  • a detoxifying-effective amount can be determined in accordance with methods known in the art .
  • 1, 4 , 7, 10 , 13 , 16-hexaazacyclohexadecane trisulfate (hexacyclen trisulfate) was purchased from Aldrich (Milwaukee, WI) and tert-butyl bromoacetate was purchased from Fluka (Ronkonkoma, NY) . All other reagents were purchased from Aldrich, Sigma (St. Louis, MO) or Fluka and were used without further purification. Chromatography was performed on silica gel 60, 220- 440 mesh ASTM (Fluka) . Thin-layer chromatography (TLC) was performed on silica gel 60 F-254 plates (EM Reagents) . All glassware used in the synthesis of the ligands after ester hydrolysis was acid-washed and rinsed with distilled deionized water.
  • X H and 13 C NMR were obtained using a Varian Gemini 300 instrument at ambient temperature. Chemical shifts were reported in ppm on the scale relative to tetramethylsilane (TMS) , trimethylsilyl propionic acid-D 5 - Na salt (TSP) or solvent. Proton chemical shifts are annotated as follows: ppm (multiplicity, integral, coupling constant (Hz) ) . Chemical ionization mass spectra (CI-MS) were obtained on a Finnegan 3000 instrument (Finnegan, San Jose, CA) . Fast atom bombardment mass spectra (FAB-MS) were obtained using an Extrel 400 instrument.
  • HR-FAB High resolution FAB mass spectra were obtained on a JEOL SX102 mass spectrometer operated at an accelerating voltage of 10 kV. Samples were desorbed from a glycerol matrix uing 6 keV xenon atoms. Mass measurements in HR-FAB were performed at 10,000 resolution. Analytical HPLC was performed on a Beckman gradient system equipped with Model 114M pumps controlled by System Gold software and a Model 165 dual wavelength detector set at 254 and 280 nm.
  • the size exclusion HPLC column (TSK-Gel® G3000SW, TosoHaas, Japan) eluted with PBS, pH 7.2, at 1 ml/min was used for these separations. Radioactive samples were counted in a Minaxi ⁇ -counter (Packard Instrument Company) .
  • HEHA N,N' ,N",N"',N"",N'""-hexaacetic acid
  • 1,4 , 7, 10, 13, 16-hexaazacyclohexadecane trisulfate was converted to the free base ([18]aneN 6 ) by neutralizing the trisulfate salt to pH > 13 with sodium hydroxide, removing water in vacuo, adding benzene, refluxing the resultant slurry with a Dean-Stark trap (Kontes Glassware Co., Vineland, NJ) for 4-6 hrs to remove residual water, filtering the benzene solution, and evaporating to dryness to yield the free base.
  • the hexaester
  • HEHA was prepared by heating the hexaester (1.1 g, 1.17 mmol) in 12 M HC1 (10 ml) at 105°C for 5 hr. The reaction mixture was cooled and the volume was reduced under vacuum until a precipitate formed. The mixture was chilled and the product was collected by filtration to yield 700 mg (73%) of HEHA as the hexahydrochloride salt. All spectra agreed with the literature values (Kimura et al., Chem Pharm Bull , 33: 655-661 (1985)). X H NMR (300 MHz, D 2 0) ⁇ 3.58 (s, 12H, N-CH 2 -CO) ; 3.34 (s, 24H, N-CH- 2 CH 2 -N) .
  • This example describes the radiolabeling of HEHA with 225 Ac.
  • Stock solutions of purified 25 Ac in 0.1 M HN0 3 were freshly prepared as needed.
  • 22S Ac was complexed with HEHA by mixing approximately 100 ⁇ l of 25 Ac solution (approx. 10 MBq, 0.1 M HN0 3 ) with 20 ⁇ l of ligand (approx. 1.0 x 10 "2 M in H 2 0) and adjusting the pH to near 5.0 by the addition of 5-10 ⁇ l of 1.0 M NH 4 OAc .
  • a Chelex column Bio-Rad Laboratories, Richmond, CA
  • This example describes the synthesis of bifunctional HEHA.
  • N- tert-butyloxycarbonyl-iminodiacetate (Boc-IDA) (20 g, 85.8 mmol, 1 eq)
  • glycine ethyl ester hydrochloride (24 g, 171.9 mmol, 2 eq)
  • triethylamine 24 ml, 172 mmol
  • DMF N, N-dimethylformamide
  • EEO l-ethyl-3- (3-dimethyl- aminopropyl) carbodiimide
  • the DMF was removed under vacuum and the residue was dissolved in ethyl acetate (600 ml) .
  • the ethyl acetate was extracted with water (200 ml x 1) ; 5% ⁇ aHC0 3 (200 ml x 3); brine (200 ml x 1) ; 1 M HCl (200 ml x 1) and brine (200 ml x 2) .
  • the organic layer was dried over Na 2 S0 4 , filtered and concentrated to yield an oil (23 g, 66%).
  • the organic layer was extracted with water (150 ml x 1) ; 5% NaHC0 3 (200 ml x 3) ; brine (200 ml x 1) ; and brine (200 ml x 2) .
  • the organic layer was dried with Na 2 S0 4 , filtered and concentrated to yield a white solid (15.86 g, 73%) .
  • the product was thoroughly dried under vacuum at 70°C prior to use in cyclization.
  • BOC-IDA-diglycine disuccinimidyl ester (5.42 g, 10 mmol) in DMF (50 ml) and N- (2-aminoethyl) -4-nitrophenylalaninamide (2.52 g, 10 mmol) in DMF (50 ml) were each loaded in 50 ml gas-tight syringes and added to the dioxane so that the addition was complete after 24 hr.
  • Second and third additions were 10 mmol and 5.5 mmol in each reactant, respectively. After the third addition, the reaction was heated for an additional 18 hr and then cooled to room temperature.
  • the reaction was concentrated to a thick brown oil under vacuum and the residue was dissolved in chloroform (500 ml) .
  • the chloroform was washed with water (1 x 200 ml) , 5 % NaHC0 3 , (2 x 200 ml), brine (1 x 200 ml), 1 M HCl ( 2 x 200 ml) , brine (1 x 200 ml) and water (1 x 100 ml) .
  • a sticky brown residue stuck to the inside of the separatory funnel was dissolved in methanol (MeOH) and reduced to dryness.
  • the CHC1 3 layer was kept separate and reduced to dryness.
  • the free base 2- (4-N0 2 -Bz- [18] ane N6) (59.3 mg, 1.51 mmol) was dissolved in anhydrous DMF (10 ml) and tert-butyl bromoacetate (1.78g, 9.1 mmol) was added. The reaction was allowed to stir in an ice-bath and then warmed to room temperature for over 1 hr. An aqueous solution of Na 2 C0 3 (965 mg in 20 ml H 2 0) was added and the mixture was stirred for 2.5 hr. Toluene (10 ml) was added and the reaction mixture was stirred overnight (17 hr) . The reaction mixture was poured into a separatory funnel and the aqueous layer was drained.
  • the toluene layer was saved.
  • the aqueous layer was extracted with CHC1 3 (2 x 100 ml) and the combined CHC1 3 and toluene layers were reduced to dryness.
  • the residue was determined to contain hexa- tert-butyl ester product by mass spectrometry .
  • the product was purified from the residue on two consecutive silica gel columns (2 x 10 cm) with a gradient from 5 - 10 % MeOH in CHC1 3 and finally 10 %
  • This example describes the attachment of a bifunctional HEHA to a targeting agent.
  • 5M NH 4 OAc buffer was passed through a column of Chelex-100 (1x7 in.) (Bio Rad, Na + form, 200-400 mesh), previously washed with water until neutral and equilibrated with the 30X buffer (200 ml) .
  • the reaction mixture is transferred to Centricon filtration units, with the appropriate MW cut-off, so that each unit contains no more than 3 mg of antibody.
  • This example describes the attachment of a bifunctional HEHA to a monoclonal antibody.
  • Antibody solutions were initially transferred to dialysis tubing (Spectra/Por CE DispoDialysers, MWCO 10K, 5 ml) and were dialyzed against conjugation buffer, (1 1, 0.05 M C0 3 "2 /HC0 3 "1 , 0.15 M NaCl, 5mM EDTA, pH 8.6) for 6 hr at 4°C.
  • conjugation buffer (1 1, 0.05 M C0 3 "2 /HC0 3 "1 , 0.15 M NaCl, 5mM EDTA, pH 8.6
  • the protein concentration was determined spectrophotometrically . Extinction coefficients of 1.4, 0.65 and 1.3 ml/mg cm were determined for mAb's BL-3, CC49 and T101, respectively, based on an mAb concentration from a protein determination using the Lowry method (Lowry et al . , J.
  • This example describes the radiolabeling of a HEHA- targeting agent with 225 Ac .
  • the pH of the radiometal solution is adjusted to 3.8-4.0 by addition of several microliters of 3 M NaOAc.
  • reaction is quenched by raising the pH to -6 with 3 M NaOAc and any free radiometal is scavenging with 5 ml of 0.5 M Na 2 EDTA solution.
  • product is purified by passage through a TSK-3000 HPLC column (Thompson Instruments) eluted with PBS at 1 ml/min.
  • This example describes the radiolabeling of the HEHA-monoclonal antibody of Example 5 with 225 AC.
  • 225 Ac (N0 3 ) 3 (2.2-4.0 mCi) was obtained from Oak Ridge National Laboratories (ORNL) as a solid. The solid was taken up in 0.1 M HCl (1 ml), such that the specific activity became 0.22-0.40 mCi/0.1 ml.
  • an aliquot of 225 Ac solution (0.05-0.2 mCi) was added to the vials containing 0.15 M NH 4 OAc (0.3- 0.4 ml) at pH 4.0, 5.0 or 7.0.
  • Preliminary labeling yields were assessed using 10 cm ITLC-SG strips (Gelman Sciences) spotted with 1 ⁇ l of the reaction and developed in 0.15 M NH 4 OAc buffer, pH 4.0. In this system the labeled proteins are retained at the origin while 225 Ac acetate moves with the solvent front. The strips were dried, cut into 1-cm segments, and counted in the ⁇ -counter. They were recounted 3 hr later in order to obtain corrected labeling yields for 2 S Ac after all of the 213 Bi that had been previously generated decayed.
  • the reaction was quenched with 0.1 M EDTA, pH 6.0 (4 ⁇ l) and 225 Ac-HEHA-mAb conjugate was purified on size exclusion HPLC column as described above. The protein fraction was collected and counted in a Capintec Radioisotope Calibrator and recounted 3 hr later to calculate the specific activity of the product.
  • the results of radiolabeling of HEHA-mAb conjugates are given in Table I. During preliminary experiments an incubation time of 30 min at 37 °C was found to produce the highest labeling yields while minimizing radiation damage to the antibody. The labeling yields for 225 AC were found to be dependent upon the pH of the reaction mixture with optimal results being obtained at pH 7.0.
  • This example describes the serum stability of the 225 Ac-radiolabeled HEHA-monoclonal antibody of Example 7.
  • Purified 25 Ac-HEHA-mAb conjugate 200 ⁇ l was incubated with fetal bovine serum (2 ml) for 3 days at 37 °C. Aliquots were withdrawn at times 0, 0.25, 1, 3, 5, 24, 48 and 72 hr and analyzed by HPLC on size exclusion column as described above. The radioactive fractions containing high and low molecular weight components were collected and counted on a ⁇ -counter immediately after elution and 3 hr thereafter in order to determine if they were due to free 225 Ac or 213 Bi .
  • the radiolabled HEHA-BL-3 construct preserved its integrity in serum for a period of at least 5 hr releasing less than 1 % of 225 Ac and 3 % of its daughter 213 Bi.
  • a peak began to grow in on the trailing slope of the 25 Ac-HEHA-BL-3 peak indicating the presence of species with molecular weight of "75-100 kDa.
  • this peak increased to be approximately one-third of the 225 Ac-HEHA-BL-3 peak.
  • This example describes the biodistribution of 225 Ac- HEHA.
  • mice Normal female BALB/c mice were injected with 2.5 ⁇ Ci in 200 ⁇ l MES buffer, pH 6.2 , in the tail vein. At 1 hr, 4 hr, 24 hr and 120 hr, 3-5 animals were sacrificed, samples of blood, liver, kidney, spleen, heart and bone were collected and weighed, and the 225 Ac content was determined. The percentage injected dose per gram (% ID/g) was calculated. The results are summarized in Table III.
  • 225 Ac-HEHA is very stable, as demonstrated by the low uptake of the radiocomplex in all tissues, including the liver.
  • 225 Ac-HEHA showed quick whole-body clearance, with essentially all of the activity excreted by 1 hour.
  • Example 10 This example describes the use of 225 Ac-HEHA- targeting agent, specifically 225 Ac-HEHA-NCS-J591 mAb, to treat cancer in vivo .
  • 225 Ac-HEHA-NCS was conjugated to the monoclonal antibody J591, which targets PSMA exc2 , an antigen that is expressed in prostate tumors and neovasculature .
  • the conjugate was administered to tumor-bearing mice.
  • conjugates of the present invention in an amount that delivered less than one atom of 225 Ac per tumor resulted in significant control of tumor growth.
  • this example demonstrates the in vivo utility of conjugates of the present invention, in the treatment of disease, such as cancer.
  • Example 11 This examples describes the attachment of HEHA, which is not bifunctional, to a protein via a carboxycarbonic anhydride.
  • a HEHA-carboxycarbonic anhydride was prepared by dissolving 50 mg HEHA (HC1) 6 in H 2 0 (1 ml) in an acid- washed 5 ml pear-shaped flask and adding triethylamine
  • a protein solution was prepared by dissolving 7.2 mg bovine albumin in 1 ml of phosphate-buffered saline solution (PBS) and switched to 50 mM bicarbonate/lOOmM NaCl with 5 mM EDTA metal-free conjugation buffer by dialysis at 4 °C.
  • PBS phosphate-buffered saline solution
  • the protein was transferred to a reaction vial (flat-bottomed plastic tube with flea bar) , the HEHA-carbonic anhydride was added, and the reaction was stirred at 4 °C for 2 hr.
  • the conjugate was initially purified by dialysis into 0.15 M ammonium acetate at 4 °C, followed by continued purification with Centricon-30 spin columns (5- 6 washes) .
  • concentration of protein conjugate was estimated from the absorbance at 280 nm (extinction coefficient of 1.4 (ml/ (cm x mg) ) .

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Molecular Biology (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

L'invention concerne un complexe chélateur à radio-isotopes émetteurs de particules α comprenant de l'actinium 225 (225Ac) et de l'acide 1,4,7,10,13,16-hexa-azacyclohexadécane-N,N',N'',N''',N'''',N'''''-hexaacétique (225Ac-HEHA). Elle concerne aussi un HEHA bifonctionnel et un 225Ac-HEHA bifonctionnel. L'acide HEHA bifonctionnel, et le 225Ac-HEHA bifonctionnel peuvent être conjugués à un agent de ciblage. Dans cette optique, l'invention a pour objet un procédé de fabrication de l'acide HEHA et des procédés de fabrication de l'acide HEHA bifonctionnel, y compris un conjugué de celui-ci. L'invention concerne aussi un procédé de traitement de maladies, un procédé de traitement du cancer, un procédé pour décontaminer un échantillon de 225Ac, un procédé pour décontaminer une personne contaminée de l'extérieur par 225Ac et un procédé de détoxication d'une personne contaminée de l'intérieur par 225Ac.
PCT/US2000/007643 1999-03-23 2000-03-23 225ac-heha et composes, procedes de synthese et procedes d'utilisation correspondants WO2000059896A1 (fr)

Priority Applications (3)

Application Number Priority Date Filing Date Title
AU44501/00A AU4450100A (en) 1999-03-23 2000-03-23 225Ac-heha and related compounds, methods of synthesis and methods of use
US09/937,030 US6696551B1 (en) 1999-03-23 2000-03-23 225Ac-HEHA and related compounds, methods of synthesis and methods of use
US10/767,133 US6995247B2 (en) 1999-03-23 2004-01-28 Ac-HEHA and related compounds, methods of synthesis and methods of use

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US12576499P 1999-03-23 1999-03-23
US60/125,764 1999-03-23

Related Child Applications (3)

Application Number Title Priority Date Filing Date
US09/937,030 A-371-Of-International US6696551B1 (en) 1999-03-23 2000-03-23 225Ac-HEHA and related compounds, methods of synthesis and methods of use
US09937030 A-371-Of-International 2000-03-23
US10/767,133 Division US6995247B2 (en) 1999-03-23 2004-01-28 Ac-HEHA and related compounds, methods of synthesis and methods of use

Publications (1)

Publication Number Publication Date
WO2000059896A1 true WO2000059896A1 (fr) 2000-10-12

Family

ID=22421309

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2000/007643 WO2000059896A1 (fr) 1999-03-23 2000-03-23 225ac-heha et composes, procedes de synthese et procedes d'utilisation correspondants

Country Status (2)

Country Link
AU (1) AU4450100A (fr)
WO (1) WO2000059896A1 (fr)

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001066155A2 (fr) * 2000-02-25 2001-09-13 Dangshe Ma Complexes et conjugat d'actinium-225 pour radio-immunotherapie
LU90544B1 (en) * 2000-03-14 2001-09-17 Europ Economic Community Bifunctional chelating agent
WO2002067999A2 (fr) * 2001-02-28 2002-09-06 Dow Global Technologies Inc. Actinium-225 complexes and conjugates for targeted radiotherapy
LU90834B1 (en) * 2001-09-17 2003-03-18 Europ Economic Community Bifunctional chelating agent for actinium
WO2020023092A3 (fr) * 2018-04-25 2020-04-23 The Johns Hopkins University Formulations de particules alpha pour le traitement de tumeurs solides
WO2022167052A1 (fr) 2021-02-08 2022-08-11 Y-Mabs Therapeutics, Inc. Utilisation d'acide ascorbique comme agent stabilisant pour des anticorps anti-b7-h3

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH03197468A (ja) * 1989-12-26 1991-08-28 Teijin Ltd 二官能性大環状キレート配位子およびその製造法
US5808003A (en) * 1989-05-26 1998-09-15 Perimmune Holdings, Inc. Polyaminocarboxylate chelators

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5808003A (en) * 1989-05-26 1998-09-15 Perimmune Holdings, Inc. Polyaminocarboxylate chelators
JPH03197468A (ja) * 1989-12-26 1991-08-28 Teijin Ltd 二官能性大環状キレート配位子およびその製造法

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
CHEMICAL ABSTRACTS, vol. 116, no. 13, 30 March 1992, Columbus, Ohio, US; abstract no. 128980a, page 898; column 1; *
G. J. BEYER ET AL: "Comparison of the biodistribution of 225-Ac and radio-lanthanides as citrate complexes", ISOTOPENPRAXIS, vol. 26, 1990, pages 111 - 114, XP000923251 *
G. J. BEYER ET AL: "The influence of EDTMP-concentration on the biodistribution of radio-lanthanides and 225-Ac in tumor-bearing mice", NUCLEAR MEDICINE & BIOLOGY, vol. 24, 1997, pages 367 - 372, XP000907686 *
K. A. DEAL ET AL: "Improved in vivo stability of actinium-225 macrocyclic complexes", JOURNAL OF MEDICINAL CHEMISTRY, vol. 42, no. 15, 29 July 1999 (1999-07-29), WASHINGTON US, pages 2988 - 2992, XP002145740 *

Cited By (14)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001066155A3 (fr) * 2000-02-25 2002-06-06 Dangshe Ma Complexes et conjugat d'actinium-225 pour radio-immunotherapie
WO2001066155A2 (fr) * 2000-02-25 2001-09-13 Dangshe Ma Complexes et conjugat d'actinium-225 pour radio-immunotherapie
LU90544B1 (en) * 2000-03-14 2001-09-17 Europ Economic Community Bifunctional chelating agent
WO2001068618A1 (fr) * 2000-03-14 2001-09-20 European Community Agent chelatant bifonctionnel
US7132513B2 (en) 2000-03-14 2006-11-07 European Community Bifunctional chelating agent
US6670456B2 (en) 2001-02-28 2003-12-30 Dow Global Technologies Inc. Actinium-225 complexes and conjugates for targeted radiotherapy
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
LU90834B1 (en) * 2001-09-17 2003-03-18 Europ Economic Community Bifunctional chelating agent for actinium
JP2005507885A (ja) * 2001-09-17 2005-03-24 ヨーロピアン コミュニティ アクチニウムのための二元機能キレート剤
US7045606B2 (en) 2001-09-17 2006-05-16 European Community Bifunctional chelating agent for actinium
WO2003024940A1 (fr) * 2001-09-17 2003-03-27 European Community Agent chelatant bifonctionnel pour l'actinium
WO2020023092A3 (fr) * 2018-04-25 2020-04-23 The Johns Hopkins University Formulations de particules alpha pour le traitement de tumeurs solides
WO2022167052A1 (fr) 2021-02-08 2022-08-11 Y-Mabs Therapeutics, Inc. Utilisation d'acide ascorbique comme agent stabilisant pour des anticorps anti-b7-h3

Also Published As

Publication number Publication date
AU4450100A (en) 2000-10-23

Similar Documents

Publication Publication Date Title
KR920003590B1 (ko) 금속 킬레이트-단백질 결합체를 형성하기 위한 다중치환된 골격을 갖는 킬레이트
KR100245941B1 (ko) 카복스아미드 변성된 폴리아민 킬레이트제 및 방사성 착화합물 및 결합체
US4994560A (en) Functionalized polyamine chelants and radioactive rhodium complexes thereof for conjugation to antibodies
US5489425A (en) Functionalized polyamine chelants
EP0587555B1 (fr) Ligand du type dtpa bifonctionnel
US5099069A (en) Backbone polysubstituted chelates for forming a metal chelate-protein conjugate
US5202451A (en) Anchimeric radiometal chelating compounds
WO1989012631A1 (fr) Agents de chelation bifonctionnels macrocycliques, leurs complexes et leurs conjugues anticorps
EP0315220A1 (fr) Méthode de formation d'un conjugué entre un complexe métallique et une protéine
EP0404605B1 (fr) Tri-aza-macrocycles et leurs procédés de préparation
IL89222A (en) Preparation of metal-radionuclide-labelled proteins, certain novel compounds used therein and radiopharmaceutical compositions containing the labelled proteins
US6696551B1 (en) 225Ac-HEHA and related compounds, methods of synthesis and methods of use
WO2000059896A1 (fr) 225ac-heha et composes, procedes de synthese et procedes d'utilisation correspondants
EP0630264A4 (fr) Ligands permettant d'ameliorer la cinetique de formation d'agents chelateurs.
EP0484989B1 (fr) Chélates de diéthylaminetriamine polysubstituée pour la formation d'une conjugaison métal-chélate-protein
WO1996011918A1 (fr) NOUVEAUX LIGANDS CHELATEURS DE TYPE N3S2 EVENTUELLEMENT RADIOMARQUES A L'AIDE DE Tc OU Re ET DESTINES A ETRE UTILISES DANS DES APPLICATIONS DIAGNOSTIQUES OU THERAPEUTIQUES
AU645258B2 (en) Radionuclide metal chelates for the radiolabeling of proteins
EP0575583A1 (fr) Complexes d'acides bicyclopolyazamacrocyclocarboxyliques, leurs conjugues, leurs procedes de preparation et leurs utilisations comme agents radiopharmaceutiques
WO1994026315A1 (fr) Complexes et conjugues de complexes d'acides bicyclopolyazamacrocyclocarboxyliques, procedes de preparation de ces derniers et d'utilisation en tant que produits radiopharmaceutiques
KR920005495B1 (ko) 다중치환된 골격을 갖는 금속 킬레이트-단백질 결합체
WO2005049096A2 (fr) Chelateurs a retention sanguine amelioree
JP2003527378A (ja) 二官能化キレート剤

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BY CA CH CN CR CU CZ DE DK DM DZ EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX NO NZ PL PT RO RU SD SE SG SI SK SL TJ TM TR TT TZ UA UG US UZ VN YU ZA ZW

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): GH GM KE LS MW SD SL SZ TZ UG ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE BF BJ CF CG CI CM GA GN GW ML MR NE SN TD TG

121 Ep: the epo has been informed by wipo that ep was designated in this application
DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
WWE Wipo information: entry into national phase

Ref document number: 09937030

Country of ref document: US

REG Reference to national code

Ref country code: DE

Ref legal event code: 8642

122 Ep: pct application non-entry in european phase
NENP Non-entry into the national phase

Ref country code: JP