WO1994025488A1 - Procede pour preparer une proteine marquee par un radionucleide de metal - Google Patents

Procede pour preparer une proteine marquee par un radionucleide de metal Download PDF

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Publication number
WO1994025488A1
WO1994025488A1 PCT/US1994/005022 US9405022W WO9425488A1 WO 1994025488 A1 WO1994025488 A1 WO 1994025488A1 US 9405022 W US9405022 W US 9405022W WO 9425488 A1 WO9425488 A1 WO 9425488A1
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Prior art keywords
protein
group
radionuclide
proteinaceous material
agent
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PCT/US1994/005022
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English (en)
Inventor
Alfons M. Verbruggen
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Mallinckrodt Medical, Inc.
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Priority to AU66708/94A priority Critical patent/AU6670894A/en
Priority to EP94915454A priority patent/EP0654043A4/fr
Publication of WO1994025488A1 publication Critical patent/WO1994025488A1/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
    • A61K51/081—Peptides, e.g. proteins, carriers being peptides, polyamino acids, proteins the protein being an albumin, e.g. human serum albumin [HSA], bovine serum albumin [BSA], ovalbumin
    • 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
    • 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
    • A61K51/088—Peptides, e.g. proteins, carriers being peptides, polyamino acids, proteins conjugates with carriers being peptides, polyamino acids or proteins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2121/00—Preparations for use in therapy
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2123/00—Preparations for testing in vivo

Definitions

  • the invention relates to a method of preparing a metal-radionuclide-labelled protein or proteinaceous material which is intended for diagnostic or therapeutic application.
  • Radionuclide-labelled compounds may be used for diagnostic examination, for example, into deviations in shape and function of internal organs and into the presence and location of pathological processes in the body.
  • a composition in which the radioactive compound is present is administered to the patient, for example, in the form of an injectable liquid.
  • suitable detection apparatus for example, a gamma camera
  • pictures of, e.g., the organ or the pathological process in which the radioactive compound has been incorporated can be obtained by recording the emitted radiation ("scanning").
  • Radioactive-labelled biological materials in particular proteins and proteinaceous materials, e.g., blood cells, serum albumin, immunoglobulins, glycopeptides, monoclonal antibodies like antimyosin and monoclonals against tumour antigens, peptides, aminofunctions-containing hormones like somatostatin and ACTH, and other proteins suitable for this purpose, such as plasmin and plasmin derivatives, e.g., miniplas in and tissue plasminogen activator, present interesting perspectives for diagnostic application.
  • proteins and proteinaceous materials e.g., blood cells, serum albumin, immunoglobulins, glycopeptides, monoclonal antibodies like antimyosin and monoclonals against tumour antigens, peptides, aminofunctions-containing hormones like somatostatin and ACTH, and other proteins suitable for this purpose, such as plasmin and plasmin derivatives, e.g., miniplas in and tissue plasminogen activator, present interesting perspectives for diagnostic application.
  • Certain proteins have a very large target organ specificity and, after having been introduced into the patient's body, can react very selectively with biological macro-molecules present therein; a good example thereof is the selective reaction of antibodies or antibody fragments with antigens present in the body.
  • Various metal-radionuclides provided they are bound to tumour-selective biological macromolecules, such as the glycopeptide bleomycin, can be used successfully for controlling tumours, and thus form a powerful tool in radiotherapy.
  • the macromolecules used thus serve as vehicles for the transportation of the desired radiation dose, i.e., the metal-radionuclide, to the tumour to be exposed to radiation.
  • the direct labelling of a protein or a proteinaceous material with a metal-radionuclide has two disadvantages.
  • the biologically active site of the protein necessary for a good target organ specificity or selectivity, may easily be blocked by this reaction, so that the normal behaviour of the biological macromolecule is disturbed.
  • the affinity between metal-radionuclide and macromolecule often is insufficient, as a result of which the formed bond is not sufficiently stable to remain intact under physiological conditions.
  • the administered material then is no longer useful, neither as diagnostic - the behaviour of the protein in the body can no longer be traced - nor as therapeutic - the radiation dose is no longer transported to the desired site but causes an undesired radiation burden elsewhere!
  • the biological behaviour of the original macromolecule must be maintained as well as possible by this modification.
  • the chelator or the bifunctional agent with which the metal-radionuclide is bound to the protein may not to be too bulky, certainly not when used for comparatively small protein molecules.
  • the usually extremely sensitive protein or proteinaceous material must be exposed as little as possible to damaging conditions during the coupling with chelator or bifunctional agent, which may adversely influence the properties of the macromolecule. Long- lasting incubations, treatments at elevated temperatures, the presence of organic solvents or conditions of acidity differing from the physiological pH, reactions in the presence of oxidizing or reducing agents, all these treatments should be avoided as much as possible.
  • the .selected bifunctional agent should ensure a strong bond between the protein or proteinaceous material on the one hand and the metal-radionuclide on the other.
  • the bond does not remain intact under physiological conditions, i.e., the radionuclide comes loose in the bloodstream and can be be transported to undesired sites in the body by other particles in the blood, the radioactive material may cause an undesired radiation burden for the tissue at those sites, and may even seriously damage the tissue there if present in therapeutically effective quantities.
  • chelators or bifunctional agents described in the above-mentioned patent publications are not satisfactory with regard to one or more of the requirements mentioned hereinbefore.
  • a comparatively bulky chelator is used in US 4479930, US 4511550, US 4652519, US 4678667 and EP 188256.
  • conditions which are damaging to the protein during the coupling reaction are applied in the methods described in US 4652440, EP 173629, WO 85/03231 and WO 86/03010.
  • the bond between protein and metal-radionuclide is not sufficiently strong in the proteins labelled according to US 4479930 and WO 85/03231.
  • SAMSA S-acetylmercapto succinic anhydride
  • This agent indeed has considerable advantages over the already known agents, because it can very readily be reacted with the protein, in which the formed protein conjugate can react with the radionuclide in one single reaction step. This latter reaction directly provides the desired labelled protein conjugate without disturbing by-products. It has been found, however, that upon labelling certain proteins according to the method described in WO 89/07456, sometimes polymerisation occurs, even under the for this reaction usual conditions which are poor in oxygen.
  • This objective can be achieved by performing the coupling reaction with a polyfunctional agent of the general formula
  • X is a halogenated or non-halogenated alkanoyl group having 2-5 carbon atoms or a substituted or non-substituted benzoyl group;
  • R is a multivalent saturated aliphatic hydrocarbyl radical, having 2-20 carbon atoms and wherein the main chain, if desired, may be interrupted by a nitrogen atom; Y is at least one terminal reactive group which is capable of reacting with a free amino group or mercapto group in the protein or the proteinaceous material; and ⁇ is 2-6; and by then, if needed after deprotection of the protected mercapto groups, reacting the formed protein conjugate with the radionuclide.
  • Metal-radionuclides suitable for use in the method according to the invention are Tc-99m, Re-186, Re-188, In-Ill, Ga-67, As-72 and As-77. Of these radionuclides Tc-99m, In-Ill, Ga-67 and As-72 may be used for diagnostic purposes, the other radionuclides are paricularly useful in therapeutically active compositions.
  • multivalent hydrocarbyl radical means a hydrocarbyl group having at least three free valencies, such as a trivalent, etc. radical.
  • the derivatisation of the protein or the proteinaceous material i.e. the reaction with the polyfunctional agent, may be carried out in a very simple manner in a neutral medium (pH between 6.5 and 8) and at room temperature.
  • the subsequent deprotection of the protected mercapto groups i.e. the cleavage of protecting groups X in the above formula I, can easily be carried out immediately following the coupling reaction, preferably by a simple treatment with hydroxylamine.
  • the desired radionuclide is presented to the protein conjugate in the form of a salt or preferably in the form of a chelate bound to comparatively weak chelators, for example, a pyrophosphate, a phosphonate or a polyphosphonate, an oxinate, a carboxylate, a hydroxycarboxylate, an aminocarboxylate, an enolate or a mixture thereof, likewise in a neutral medium.
  • the desired complex is formed via the principle of ligand exchange, in which the sulphur atom of the thio compound forms a strong chelate bond with the metal-radionuclide.
  • a mercapto-protecting group X is very important because for that reason polymerisation of the protein conjugate during the preparation is avoided.
  • suitable protecting groups X for the mercapto group are: acetyl, halogenated acetyl and substituted or non-substituted benzoyl, in which in particular electrons-withdrawing groups, such as nitro, halogen and sulpho are to be considered as suitable substituents.
  • a suitable protein to be labelled by using the method of the present invention is albumin, e.g. human serum albumin (HSA) . It has been found, that HSA labelled with Tc-99m via the method of the invention, i.e.
  • the present invention also relates to the protein conjugate complexed with the radionuclide, as mentioned above, and to the protein conjugate per se, prepared as described above by reacting a protein or proteinaceous material with the polyfunctional coupling agent of formula I.
  • Y' is an isocyanate group, a for yl group, a diazonium group, an isothiocyanato group, an epoxyethylene group, a trichloro-s-triazinyl group, an ethylene i ino group, a halocarbonyl group, a halosulphonyl group, a malei ido group, a sulphonated or non-sulphonated alkylcarbonyloxycarbonyl group, a sulphonated or non-sulphonated alkylcarbonyliminocarbonyl group, a 2,4-dinitrophenoxycarbonyl group, or a sulphonated or non-sulphonated nitrogen-containing heterocyclic five- or six-membered ring which is bound to R with the ring nitrogen via a carbonyl group or oxycarbonyl group and which is substituted in the ortho position with an oxo function or a thioxo function.
  • alkylcarbonyloxycarbonyl groups and of alkylcarbonyliminocarbonyl groups are radicals derived from succinic anhydride and succinimide, respectively.
  • suitable protecting groups X are: acetyl, halogenated acetyl such as trifluoroacetyl, and benzoyl whether or not substituted with nitro, (C 1 -C 4 )alkyl, (C 1 -C 4 ) alkoxy, halogen or sulpho.
  • Y' a 2,4-dinitro-phenoxycarbonyl group or a sulphonated or non-sulphonated nitrogen-containing heterocyclic five- or six-membered ring which is bound to R with the ring-nitrogen via a carbonyl group or oxycarbonyl group and which is substituted in the ortho position with an oxo function or a thioxo function.
  • Y is a sulphonated or non-sulphonated succinimido-oxy group or a 2-thioxo-thiazolidin-3-yl group.
  • a succinimido-oxy group is also called a
  • R- is hydrogen or C ⁇ -C. 4 alkyl
  • R 2 is C 0 -C 3 alkylene
  • R 3 is C ! -C 5 alkylene
  • R 4 , R 5 and R 6 are each individually straight or branched Cx-C ⁇ alkylene
  • the last-mentioned preferred agents are excellently suitable for use in the preparation of labelled proteins or proteinaceous materials according to the present invention. It has been found in addition that the last-mentioned polyfunctional agents can react with the amino functions of the protein selectively and without damage to the protein molecule.
  • Specific examples of trifunctional agents included in the last-mentioned general formula IV, sub (1), may be presented by the following formulas:
  • Z has the meaning given hereinbefore, and A 2 is hydrogen or an alkali- (or ammonium-)sulphonate group.
  • the invention also relates to a metal-radionuclide-labelled protein or proteinaceous material obtained by using the method as described hereinbefore, and to a radiopharmaceutical composition which comprises, in addition to a pharamaceutically acceptable liquid carrier material, a metal-radionuclide-labelled protein or proteinaceous material.
  • a radiopharmaceutical composition which comprises, in addition to a pharamaceutically acceptable liquid carrier material, a metal-radionuclide-labelled protein or proteinaceous material.
  • the resulting solution of the labelled protein or proteinaceous material may be used directly as a radiopharmaceutical composition. If necessary, the solution may be brought into a form which is better suitable for intravenous or subcutaneous administration, for example, by the addition of a pharmaceutically acceptable liquid carrier material, preferably a physiological saline solution.
  • a pharmaceutically acceptable liquid carrier material preferably a physiological saline solution.
  • the composition for carrying out a radiodiagnostic examination the composition, as described hereinbefore, optionally after dilution with a pharmaceutically acceptable liquid, preferably a physiological saline solution, may be administered to a warmblooded living being in a quantity from 100 ⁇ Ci to 30 mCi, preferably from 0.5 to 10 mCi, per 70 kg of body weight, after which the radioactive radiation emitted by the living being is recorded. If the composition is to be used for a radiotherapeutic treatment, a suitable metal-radionuclide should be selected for the labelling reaction, as indicated hereinbefore. Upon use, the composition, optionally after dilution with a pharmaceutically acceptable liquid, is administered to a warmblooded living being in a quantity effective for combating or controlling tumours.
  • a pharmaceutically acceptable liquid preferably a physiological saline solution
  • the invention therefore also relates to a so-called "kit", comprising (1) in an optionally dry condition a composition of a protein conjugate, which is formed by reaction of a protein or a proteinaceous material with a polyfunctional agent as defined hereinbefore, (2) a solution of a salt or chelate of a metal-radionuclide, and (3) instructions for use with a prescription for reacting the ingredients present in the kit.
  • a protein hereafter is to be understood to mean a protein including a proteineous material.
  • the desired radionuclide is preferably presented to the protein conjugate in the form of a chelate bound to comparatively weak chelators, for example, a pyrophosphate, and phosphonate or polyphosphonate, an oxinate, a carboxylate, a hydroxycarboxylate, an aminocarboxylate, an enolate or a mixture thereof, in which the reaction can take place in a neutral medium.
  • a chelate bound to comparatively weak chelators for example, a pyrophosphate, and phosphonate or polyphosphonate, an oxinate, a carboxylate, a hydroxycarboxylate, an aminocarboxylate, an enolate or a mixture thereof, in which the reaction can take place in a neutral medium.
  • Suitable chelators for the radionuclide are 8-hydroxyquinoline or derivatives thereof; dicarboxylic acids, polycarboxylic acids or hydroxycarboxylic acids, for example, oxalic acid, malonic acid, succinic acid, maleic acid, orthophthalic acid, malic acid, lactic acid, tartaric acid, citric acid, ascorbic acid, salicylic acid or derivatives of these acids; pyrophosphates; phosphonates or polyphosphonates, for example, methylene diphosphonate, hydroxyethylene diphosphonate or hydroxymethylene diphosphonate; or enolates, for example with a B-diketone such as acetyl acetone, furoyl acetone, thenoyl acetone, benzoyl acetone, dibenzoyl methane, tropolone or derivatives of these diketones.
  • a B-diketone such as acetyl acetone, furoyl acetone, theno
  • 8-Hydroxyquinoline, citric acid, tartaric acid, ascorbic acid, glucoheptonic acid or a derivative thereof, or acetyl acetone are to be considered as particularly suitable as chelators, because it has been found that a chelate of a radionuclide with one of these chelators in a suitable medium, preferably a buffered aqeuous solution, easily reacts at a physiological pH with a protein conjugate as defined hereinbefore, the desired radionuclide complex being formed by ligand exchange in a high yield and purity.
  • the supplied kit may also comprise the constituents mentioned sub (1) with instructions for use, whereas the solution of the metal- radionuclide defined sub (2), having a limited shelf life, may be supplied to the user separately.
  • the kit according to the invention is equipped so as to comprise the following ingredients: (1) in an optionally dry condition a composition of a protein conjugate, which is formed by reaction of a protein with a polyfunctional agent as defined hereinbefore; (2) a chelator as described hereinbefore and a reducing agent; and (3) instructions for use with a prescription for reacting the ingredients of the kit with technetium-9 m in the form of a pertechnetate solution.
  • the composition should comprise a reducing agent to reduce the pertechnetate, for example, a dithionite or stannous ions.
  • a kit is intended for the preparation of a Tc-99m-labelled pharmaceutical composition.
  • the pertechnetate solution can simply be obtained by the user from a molybdenum-technetium generator available to him.
  • a similar kit may be used for the preparation of a pharmaceutical composition labelled with Re-186 or Re-188, in which the perrhenate solution must also be reduced with a suitable reducing agent, for example, a dithionite or stannous ions.
  • a suitable reducing agent for example, a dithionite or stannous ions.
  • the ingredients defined above sub (1) and (2) may be combined, provided they are compatible.
  • Such a kit, in which the combined ingredients are preferably lyophilized, is extremely suitable for being reacted by the user with the radionuclide solution in a simple manner.
  • the kit according to the invention is equipped so as to comprise (1) in an optionally dry condition a polyfunctional agent as defined hereinbefore, as well as a chelator as described hereinbefore and a reducing agent, and (2) instructions for use with a prescription for reacting the ingredients mentioned sub (1), which are preferably accomodated in one vial, with a protein which is separately supplied to the user, and then with technetium-99m in the form of a pertechnetate solution or with rhenium-186 or rhenium-188 in the form of a perrhenate solution.
  • a polyfunctional agent as defined hereinbefore
  • a chelator as described hereinbefore and a reducing agent
  • the kit according to the invention comprises (1) a polyfunctional agent as defined hereinbefore, (2) a solution of a salt or chelate of a metal-radionuclide, and (3) instructions for use with a prescription for reacting the ingredient sub (1) with a protein and then with the ingredient mentioned sub 2.
  • a metallic reducing agent for example, Sn(II),
  • Fe(II), Cu(I), Ti(III) or Sb(III), is preferably used as a reducing agent for the kits mentioned hereinbefore; Sn(II) is excellently suitable.
  • the constituent of the above-mentioned kits stated sub (1) may be supplied as a solution, for example, in the form of a physiological saline solution, or in some buffer solution or other, but is preferably present in a dry condition, for example, in a lyophilized condition.
  • a component for an injection liquid it should be sterile, in which, if the constituent is present in a dry condition, the user should use a sterile physiological saline solution as a solvent.
  • the above-mentioned constituent may be stabilised in the usual manner with suitable stabilisers, or may comprise other auxiliary means like fillers, e.g, glucose, lactose, mannitol, and the like.
  • 2,3- (di-S-acetylmercapto)propionic acid is prepared by reaction of 2,3-dibromopropionic acid and ercaptoacetic acid according to Ondetti et al. (German pat. appln. 2,752,720). To a solution of 2,3-(di-S-acetylmercapto)propionic acid is prepared by reaction of 2,3-dibromopropionic acid and ercaptoacetic acid according to Ondetti et al. (German pat. appln. 2,752,720). To a solution of 2,3-(di-(di-(di-)
  • Example II Coupling of albumin with SATP
  • a solution containing varying amounts (3-30 umol) of SATP in DMSO is added 10 ⁇ l of a solution containing varying amounts (3-30 umol) of SATP in DMSO.
  • the unreacted ligand is removed by chromatography over a Sephadex G25M column; swelling in saline and elution with a 0.05 M phosphate buffer pH 7.5 containing ImM EDTA.
  • the fractions containing the albumin are combined.
  • Deacetylation of the S-acetyl protected mercapto groups is carried out by incubation for 2 h. after addition of 0.2 ml of a deacetylation mixture (50 mM sodium phosphate, 25 mM EDTA, 0.5 M hydroxylamine, pH 7.5).
  • a deacetylation mixture 50 mM sodium phosphate, 25 mM EDTA, 0.5 M hydroxylamine, pH 7.5.
  • An additional purification can be carried out by size exclusion HPLC to remove hydroxylamine, using a Biosil® SEC 250 column; elution with phosphate buffer pH 7. The peak with a similar retention time as native serum albumin is collected and used for labelling.
  • Example III Labelling of SATP-modified albumin with technetium-99m Direct labelling with 99 ⁇ Tc is performed by addition of 10 ⁇ g of SnCl 2 .2H 2 0 dissolved in 5 ⁇ l HCL 0.05 N and 1 ml eluate of a commercial Tc-generator containing 0.55 GBq 99m Tc in the form of sodium pertechnetate to the HPLC-purified fraction containing the deacylated modified albumin. After incubation for a few min, the labelling reaction mixture is analyzed and purified by the above SEC-HPLC. the column effluent is monitored for both UV absorbance at 280 nm and radioactivity. The peak with a retention time similar to that of albumin is isolated.
  • Exchange labelling is performed by mixing 2 ml of the deacylated modified albumin solution with 0.5 ml of a 99m Tc-gluconate solution and incubation for 20 min at 37°C.
  • 99m Tc-gluconate 20 mg of sodium gluconate is dissolved in 1 ml of 0.5 M phosphate buffer pH 7 and 100 ⁇ g SnCl 2 .2H 2 0 in 25 ⁇ l HC1 O.05N is added, followed by 1 ml of 99m Tc-pertechnetate solution containing 0.55 GBq 99m Tc. Analysis and purification are performed as for the direct labelling reaction mixture.
  • the product is indicated in the following Examples as 99mTc-DMP-HSA.
  • the radiolabelled preparations are diluted with saline to a concentration of 3.7 MBq/ml and 125 I-albumin is added to a concentration of 0.37 MBq/ml.
  • a male albino rabbit (2.5-3.1 kg) is sedated and 0.5 ml of the diluted tracer solution is injected via an ear vein.
  • a 2 ml blood sample is taken from the artery of the contralateral ear.
  • the 125 I- and 99m Tc-activity are measured as described above and compared with the activity of a standard solution of the injected preparation.
  • the percentage of injected activity in 100 g of blood is calculated at each time interval and also the ratio between the percentage of injected 99m Tc-activity remaining in the plasma and the corresponding percentage of injected 125 I-activity at the same time interval.
  • 99m Tc-DMP-albumin as well as a common 99m Tc-labelled HSA preparation (Technescan® HSA) are compared with a one week time interval in a healthy male volunteer.
  • the radiolabelled preparations are diluted with saline to a concentration of 3.7 MBq/ml and 0.5 ml is injected via an arm vein.
  • a 2-ml blood sample is taken from the contralateral arm and the 99m Tc-activity is measured as described above. Data are related to the activity in the blood at 5 min p.i., which allows to compare the plasma disappearance of the preparations tested.
  • the volunteer is asked to void at 30 min, 60 min and 120 min a.i., and the percentage of injected dose excreted at these time intervals is determined without correction for the postvoid residual urine.
  • the blood disappearance curves obtained are shown in figure 2.
  • the activity in the blood at 5 min p.i., i.e. after sufficient mixing of the injected substances with the blood, is equalled to 100% and the activities at the other time intervals are related to this value.
  • 99mTc-DMP-albumin albumin deivatized with SATP in a 1:25 ratio
  • the blood activity of 99m Tc-HSA decreases continuously during the 2 hours of the investigation. After 2 hours, only 60% of the initial activity is retained within the blood.

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Abstract

L'invention se rapporte à un procédé pour préparer une protéine ou une matière protéinée marquée par un radionucléide de métal, la protéine ou la matière protéinée étant destinée à des applications diagnostiques ou thérapeutiques. Ce procédé consiste à faire réagir une protéine ou une matière protéinée avec un agent bifonctionnel pour coupler le radionucléide à la protéine ou à la matière protéinée, un conjugué de protéine étant formé par réaction entre l'agent bifonctionnel et les groupes amino ou mercapto libres de la protéine ou de la matière protéinée, puis en complexant le radionucléide avec le conjugué ainsi formé pour obtenir un complexe de radionucléide. L'invention se rapporte en outre à la protéine ou matière protéinée marquée ainsi obtenue, ainsi qu'à une trousse pour préparer une composition radiopharmaceutique, cette trousse comprenant l'agent polyfonctionnel mentionné ci-dessus ou un conjugué de protéine formé par réaction d'une protéine ou matière protéinée avec ledit agent polyfonctionnel.
PCT/US1994/005022 1993-05-03 1994-05-02 Procede pour preparer une proteine marquee par un radionucleide de metal WO1994025488A1 (fr)

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Application Number Priority Date Filing Date Title
AU66708/94A AU6670894A (en) 1993-05-03 1994-05-02 Method for preparing a metal-radionuclide-labelled protein
EP94915454A EP0654043A4 (fr) 1993-05-03 1994-05-02 Procede pour preparer une proteine marquee par un radionucleide de metal.

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EP93201249 1993-05-03

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CA (1) CA2141739A1 (fr)
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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5746996A (en) * 1994-06-03 1998-05-05 Immunomedics, Inc. Thiolation of peptides for radionuclide-based radiodetection and radiotherapy
US5941913A (en) * 1996-10-10 1999-08-24 Chas. A Blatchford & Sons Limited Above-knee lower limb prosthesis and a shin component for the prosthesis

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4479930A (en) * 1982-07-26 1984-10-30 Trustees Of The University Of Massachusetts Amines coupled wth dicyclic dianhydrides capable of being radiolabeled product
EP0237150A2 (fr) * 1986-03-12 1987-09-16 Neorx Corporation Couplage de radionuclide et d'anticorps
US4732974A (en) * 1986-03-05 1988-03-22 Mallinckrodt, Inc. Metal ion labeling of carrier molecules
WO1989007456A1 (fr) * 1988-02-09 1989-08-24 Mallinckrodt, Inc. Procede de preparation d'une proteine marquee par un nucleide radio-metallique
WO1991007991A1 (fr) * 1989-11-30 1991-06-13 Mallinckrodt Medical, Inc. Procede de preparation d'une proteine marquee par un radionuclide de metal

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4116962A (en) * 1976-12-03 1978-09-26 E. R. Squibb & Sons, Inc. Pyrrolidine and piperidine-2-carboxylic acid derivatives
US5218128A (en) * 1988-06-15 1993-06-08 Centocor, Inc. Bifunctional coupling agents and radionuclide labeled compositions prepared therefrom
US5162505A (en) * 1989-09-19 1992-11-10 Centocor Proteins modified with positively charged carriers and compositions prepared therefrom
DE4033714C3 (de) * 1990-10-24 1995-09-07 Brahms Diagnostica Gmbh Neues Hapten-Analytikum basierend auf dem Prinzip einer Antigen-Antikörper-Reaktion

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4479930A (en) * 1982-07-26 1984-10-30 Trustees Of The University Of Massachusetts Amines coupled wth dicyclic dianhydrides capable of being radiolabeled product
US4732974A (en) * 1986-03-05 1988-03-22 Mallinckrodt, Inc. Metal ion labeling of carrier molecules
EP0237150A2 (fr) * 1986-03-12 1987-09-16 Neorx Corporation Couplage de radionuclide et d'anticorps
WO1989007456A1 (fr) * 1988-02-09 1989-08-24 Mallinckrodt, Inc. Procede de preparation d'une proteine marquee par un nucleide radio-metallique
WO1991007991A1 (fr) * 1989-11-30 1991-06-13 Mallinckrodt Medical, Inc. Procede de preparation d'une proteine marquee par un radionuclide de metal

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
European Journal of Nuclear Medicine, Volume 20, No. 6, issued June 1993, K.A. VERBEKE et al., "Technetium-99m Mercaptoalbumin as a Potential Substitute for Technetium-99m Labelled Red Blood Cells", pages 473-482, especially page 473. *
See also references of EP0654043A4 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5746996A (en) * 1994-06-03 1998-05-05 Immunomedics, Inc. Thiolation of peptides for radionuclide-based radiodetection and radiotherapy
US5941913A (en) * 1996-10-10 1999-08-24 Chas. A Blatchford & Sons Limited Above-knee lower limb prosthesis and a shin component for the prosthesis

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EP0654043A4 (fr) 1997-07-16
CA2141739A1 (fr) 1994-11-10
EP0654043A1 (fr) 1995-05-24
AU6670894A (en) 1994-11-21
MXPA94003278A (es) 2005-04-01

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