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WO1997028181A2 - Stabilisation apres marquage de proteines et de peptides radiomarquees - Google Patents

Stabilisation apres marquage de proteines et de peptides radiomarquees Download PDF

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Publication number
WO1997028181A2
WO1997028181A2 PCT/US1997/001695 US9701695W WO9728181A2 WO 1997028181 A2 WO1997028181 A2 WO 1997028181A2 US 9701695 W US9701695 W US 9701695W WO 9728181 A2 WO9728181 A2 WO 9728181A2
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WO
WIPO (PCT)
Prior art keywords
radiolabeled
protein
peptide
ascorbic acid
stabilizer
Prior art date
Application number
PCT/US1997/001695
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English (en)
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WO1997028181A9 (fr
WO1997028181A3 (fr
Inventor
Paul O. Zamora
Michael J. Marek
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Rhomed Incorporated
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Application filed by Rhomed Incorporated filed Critical Rhomed Incorporated
Priority to CA002279349A priority Critical patent/CA2279349C/fr
Priority to AU25264/97A priority patent/AU2526497A/en
Publication of WO1997028181A2 publication Critical patent/WO1997028181A2/fr
Publication of WO1997028181A3 publication Critical patent/WO1997028181A3/fr
Publication of WO1997028181A9 publication Critical patent/WO1997028181A9/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
    • 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

Definitions

  • This invention relates to stabilizers for radiopharmaceutical compositions which are added after radiolabeling but prior to admimstration.
  • Ascorbic acid and/or a derivative thereof, alone or in combination with other stabilizers, is used to inhibit oxidation loss and autoradiolysis of radiolabeled peptides and proteins.
  • Protein- or peptide-based radiopharmaceuticals are primarily based upon use of monoclonal antibodies (or fragments thereof) as a targeting vehicle, but other peptides or proteins can also be used, including albumins and hormones. Both intact antibodies (monoclonal and polyclonal) and fragments, made by any method known to the art, as well as peptide mimics of fragments or antibody binding sites can be radiolabeled and used as imaging, diagnostic or therapeutic agents.
  • Radiolabeled peptide analogues of somatostatin used for diagnostic imaging include 123 I-labeled Tyr-3-octreotide and n, In-diethylene tetraaminepentaacetic acid (DTPA)-octreotide imaging agents.
  • DTPA In-diethylene tetraaminepentaacetic acid
  • Both protein- and peptide-based radiopharmaceuticals may be radiolabeled by a variety of means. Both peptides and proteins can be directly radioiodinated, through electrophilic substitution at reactive aromatic amino acids. Iodination may also be accomplished via prelabeled reagents, in which the reagent is iodinated and purified, and then linked to the peptide or protein.
  • the utility of DTPA and EDTA chelates covalently coupled to proteins, polypeptides and peptides is well known in the art.
  • DTPA has been used as a bifiinctional chelating agent for radiolabeling a variety of peptides with m In, including somatostatin analogues for cancer imaging, ⁇ -melanocyte-simulating hormone for imaging melanoma, chemotactic peptides for infection imaging, laminin fragments for targeting tumor-associated laminin receptors and atrial natriuretic peptides for imaging atrial natriuretic receptors in the kidney.
  • Tc is a preferred isotope for diagnostic imaging, due to its low cost, ready availability, excellent imaging properties and high specific activities.
  • Two approaches have been described for radiolabeling proteins and peptides with ""Tc: direct labeling and bifiinctional chelates.
  • Direct labeling methods are generally described in U.S. Patents 5,078,985; 5,102,990; 5,277,893; 5,443,816; and 5,460,785 referenced above, in which a variety of methods of direct labeling of peptides and proteins through sulfur-, oxygen- and nitrogen-containing amino acid sequences available for binding are disclosed.
  • a variety of high affinity chelates to bind Tc to specific sites on peptides have been developed.
  • the bifiinctional reagent is first labeled with ""Tc, and then conjugated to the peptide.
  • a chelating agent is covalently attached to the peptide prior to radiolabeling.
  • Chelates which have been employed include a variety of N2S2 and N3S ligands, DTPA, 6-hydrazinonicotinate groups, metallothionein and metallothionein fragments.
  • Isotopes of rhenium principally ,86 Re and 188 Re, have been used to radiolabel proteins and peptides for investigation as therapeutic agents.
  • the chemistry of 1M Re and I88 Re is similar to that of ""Tc, though not identical, and both direct and chelate labeling approaches have been used in radiolabeling proteins and peptides with rhenium.
  • Radiopharmaceutical compositions are known to degrade after radiolabeling, primarily by oxidation losses and by autoradiolysis. Some radiopharmaceuticals, such as Tc, and especially 186 Re and 188 Re labeled compounds, are particularly susceptible to oxidation losses if the isotope is not maintained in a suitable oxidation state. Both technetium and rhenium isotopes normally exist in their highest or +1 oxidation state, which is the stable state, until reduced with stannous or other reducing agents.
  • a technetium or rhenium radiolabeled compound can become unstable if the complexed reduced isotope is oxidized to a higher oxidation state, releasing the bound isotope as free or unbound pertechnetate +7 or free perrhenate +7.
  • Autoradiolysis includes chemical decomposition of a radiolabeled peptide or protein by the action of radiation emitted from the radioisotope coupled to the peptide or protein. Autoradiolysis may be caused by the formation of free radicals in the water or other medium due to the effect of radiation emitted from the radioisotope. Free radicals are molecules or atoms containing a single unpaired electron, which exhibit high chemical reactivity. Autoradiolysis is a significant problem with high energy Remitting isotopes, such as rhenium isotopes, and with ⁇ -emitting isotopes, but is typically somewhat less of a problem with ⁇ -emitting isotopes, such as ""Tc.
  • Patent 5,384,113 incorporated herein by reference, and use of ascorbic acid to stabilize some chemical-based radiolabeled compounds, but not protein- or peptide-based radiolabeled compounds, is described in Tofe, AJ. and Francis, M.D., J. Nucl. Med., 17, 820-825
  • a composition containing a radiolabeled protein or peptide having improved stability e.g., against oxidation, autoradiolysis and more generally degradation.
  • Another object is to provide novel methods for stabilizing a radiolabeled protein peptide or polypeptide.
  • the present invention provides a method for stabilizing radiopharmaceutical compositions, including compositions based on peptides and proteins, including antibodies.
  • the method comprises adding ascorbate or other stabilizing agents to the pre-radiolabeled composition in an amount effective to prevent degradation of the radiolabeled substance, such as that caused by autoradiolysis of the labeled composition and oxidation loss of the radiolabel.
  • a method of preparing a stabilized peptide or protein radiopharmaceutical composition comprising the ordered steps of labeling a peptide or protein with a radioisotope to form a radiolabeled pharmaceutical product, said radiolabeled pharmaceutical product being substantially free of any stabilizing agents, and then adding a stabilizing agent consisting of ascorbic acid, or one or more derivatives thereof (such as salts, esters, and mixtures thereof) to the radiolabeled pharmaceutical product.
  • the radioisotope may be an isotope of rhenium or technetium, and particularly ""Tc, 186 Re or 188 Re.
  • a method of preparing a stabilized rhenium-labeled peptide-based somatostatin analogue radiopharmaceutical composition comprising the ordered steps of labeling said peptide with an isotope of rhenium to form a radiolabeled pharmaceutical product, said radiolabeled pharmaceutical product being heretofore substantially free of any stabilizing agents, and then adding a stabilizing agent consisting of ascorbic acid, its salts, esters, derivatives and mixtures thereof to the radiolabeled pharmaceutical product.
  • the radioisotope may be 186 Re or 188 Re.
  • a method of preparing a stabilized technetium-labeled anti-SSEA-1 IgM antibody-based radiopharmaceutical composition comprising the ordered steps of labeling said anti-SSEA-1 antibody with an isotope of technetium to form a radiolabeled pharmaceutical product, said radiolabeled pharmaceutical product being heretofore substantially free of any stabilizing agents, and then adding a stabilizing agent consisting of ascorbic acid, its salts, esters, derivatives and mixtures thereof to the radiolabeled pharmaceutical product.
  • the radioisotope may be ""Tc.
  • the present invention provides stabilized compositions containing a radiolabeled protein or peptide, which compositions have been stabilized by the addition of a stabilizing agent containing ascorbic acid or a derivative thereof to a composition containing said protein or peptide already radiolabeled.
  • FIGURE 1 Elution profile of Re-188-RC-160 labeled with 65 mCi of Re-188 at 6 hours post-labeling from a C 18 -reverse-phase HPLC column.
  • the y-axis in arbitrary units.
  • the figure inset is an elution profile of Re-188-RC- 160 from a C, g -SepPak column using a step-gradient of acidified ethanol.
  • FIGURE 2 Comparative elutions profiles of Re-188-RC-160 radiolabeled at either 90°C or 37°C.
  • FIGURE 3 Displacement of Re-188 from Re-188-RC-160 using increasing concentrations of cysteine as a challenge agent.
  • FIGURE 4 Comparative biodistribution of Re-188-RC-160 radiolabeled at either 90°C or 37°C in normal female mice.
  • FIGURE 5 Comparison of Tc-99m- or Re-188- labeled RC-160 over time as analyzed by TLC. Saline was used as a mobile phase. Similar amounts (20 mCi) of each radionuclide were used, and the same formulation (optimized for Re-labeling) was used in the comparison. No post- labeling stabilizer was used.
  • FIGURE 6 Comparative TLC of Re-188-RC-160 with and without post-labeling stabilization with ascorbate.
  • HGURE 7 Elution profile of Re-188-RC-160 labeled with 65 mCi of Re-188 at 30 hours post-labeling from a C -reverse-phase HPLC column. The y-axis in arbitrary units.
  • HGURE 8 Comparative elution profiles of Re-188-RC-160 with ascorbate added after the radiolabeling (top) or before the radiolabeling (bottom). Both preparations were radiolabeled with 10 mCi (370 Mbq) of Re-188.
  • HGURE 9 Labeling of a RhoMed radiophamaceutical produ . known as LeuTec- MTM, a ""Tc-labeled anti-SSEA- 1 antibody. (This product is referred to as "Leuko- 1" in Figure 9, which is a summary of the test results from a lot of LeuTecMTM manufactured by RhoMed.) This data shows the difference in radiolabeling yields between unstabilized and stabilized (with ascorbate) product.
  • HGURE 10 HPLC profile on Tc-labeled IgM samples showing the difference in yield over time with labeling occurring in the presence of ascorbate or with postlabeling addition of ascorbate.
  • the present invention is directed to radiolabeled protein- or peptide-containing compositions with a stabilizing agent added subsequent to radiolabeling but prior to use to prevent oxidation and autoradiolysis.
  • a stabilizing agent which is effective at preventing oxidation and autoradiolysis is ascorbic acid (and derivatives thereof).
  • ascorbic acid and its derivatives interfere with radiolabeling if included in the composition prior to or during radiolabeling.
  • the inventors have found that, unexpectedly, when added subsequent to radiolabeling (and subsequent to any incubation period), ascorbic acid and its derivatives result in a radiolabeled substance of superior stability and body (volume).
  • radiolabeled proteins and peptides which are effectively stabilized according to the present invention include radiopharmaceutical drugs having diagnostic or therapeutic applications. While the methods of this invention are particularly applicable to stabilizing compositions including isotopes of technetium and rhenium, such as ""Tc,
  • compositions including a wide variety of isotopes including those found in the group consisting of elements 26-30 (Fe, Co, Ni, Cu, Zn), 33-34 (As, Se), 42-50 (Mo, Tc, Ru, Rh, Pd, Ag, Cd, In, Sn) and 75-85 (Re, Os, Ir, Pt, Au, Hg, Tl, Pb, Bi, Po, At) of the group consisting of elements 26-30 (Fe, Co, Ni, Cu, Zn), 33-34 (As, Se), 42-50 (Mo, Tc, Ru, Rh, Pd, Ag, Cd, In, Sn) and 75-85 (Re, Os, Ir, Pt, Au, Hg, Tl, Pb, Bi, Po, At) of the group consisting of elements 26-30 (Fe, Co, Ni, Cu, Zn), 33-34 (As, Se), 42-50 (Mo, Tc, Ru, Rh, Pd, Ag, Cd, In, Sn) and
  • Radioisotopes with diagnostic or therapeutic applications include 62Cu, 64Cu, 67Cu, 97Ru, 105Rh, 109Pd, ,86 Re, l88 Re, 198Au, 199Au, 203Pb, 211Pb and 212Bi.
  • radiolabeled peptide compositions stabilized by means of this invention include proteins, peptides and polypeptides which are naturally-occurring, as well as those produced by chemical synthesis, by recombinant DNA technology, by biochemical or enzymatic fragmentation of larger molecules, or by any other means for producing them.
  • peptides stabilized by the present method include peptide fragments, polypeptides and other structures derived therefrom, generally consisting of a sequence of amino acids.
  • peptides include those derived from laminin, fibronectin, cytokines, lymphokines, serum albumin, fibrinogen, enzymes, hormones, somatostatin, urokinase, tissue plasminogen activator, and protease inhibitors.
  • Peptides will generally comprise fewer than 100 amino acids, and preferably fewer than 60 amino acids, and most preferably comprise between about 4 and about 20 amino acids.
  • amino acids forming all or a part of the peptide may be naturally occurring amino acids, isomers and modifications of such amino acids, non-protein amino acids, post-translationally modified amino acids, enzymatically synthesized amino acids, isosteric amino acid analogues and the like, so that the term “peptide” includes pseudopeptides and peptidomimetics.
  • peptide also includes cyclic peptides, bridged peptides (e.g. , disulfide-bridged peptides), dimers or multimers of peptides.
  • Radiolabeled protein compositions stabilized by means of this invention include such proteins, whether natural or synthetic, as human serum albumin, fibrinogen, urokinase, gamma globulin, laminin, fibronectin, cytokines, lymphokines, enzymes, enzyme inhibitors, hormones, glycoproteins, and immunoglobulins.
  • the term "protein” as used throughout the specification and claims is intended to include all of the foregoing substances.
  • the protein is typically of mammalian origin, but also includes proteins of plant origin and proteins from prokaryotic cells. Methods of attaching or complexing proteins to other molecules, such as lipids and carbohydrates, including liposomes, are known to those skilled in the art.
  • Immunoglobulins include antibodies and antibody fragments (including fragments consisting essentially of an antigenic determinant or antigen binding site), of any species, and include both polyclonal and monoclonal antibodies made by any means, as well as chimeric and genetically engineered antibodies, hybrids, and fragments of all of the foregoing.
  • immunoglobulins of any class such as IgG, IgM, IgA, IgD or IgE, of any species origin, including human beings, chimeric antibodies or hybrid antibodies with dual or multiple antigen or epitope specificities, and fragments of all of the foregoing, including F(ab)2, F(ab)2, Fab, Fab and other fragments, including hybrid fragments, and further includes any immunoglobulin or any natural, synthetic or genetically engineered protein that functionally acts like an antibody by binding specifically to a given antigen to form a complex, including single chain antibodies.
  • antibody or “antibodies” as used throughout the specification and claims are intended to include all such antibodies and antibody fragments.
  • the products stabilized by means of the invention set forth herein can be used for both medical applications and veterinary applications. Typically, the product is used in humans, but may also be used in other mammals.
  • the term "patient” is intended to include a mammal, and is so used throughout the specification and in the claims.
  • the primary applications of the invention involve human patients, but the invention may be applied to laboratory, farm, zoo, wildlife, pet or sport animals.
  • the products stabilized by means of the invention set forth herein can be used as imaging agents, for example, to view tissues in order to monitor normal or abnormal metabolic events, to localize normal or abnormal tissues, to localize diseases, to diagnose or treat diseases, and to bind to blood constituents, including blood cells, such as lymphocytes, for subsequent localization of diseases, infections, and abnormal tissues.
  • imaging agents for example, to view tissues in order to monitor normal or abnormal metabolic events, to localize normal or abnormal tissues, to localize diseases, to diagnose or treat diseases, and to bind to blood constituents, including blood cells, such as lymphocytes, for subsequent localization of diseases, infections, and abnormal tissues.
  • the application and medical use of the product depends on the type of protein or peptide and the type of radioisotope. used.
  • the protein or peptide is first labeled with a radioisotope which can be accomplished using known techniques, and once labeling is complete, it is then stabilized according to the invention.
  • the radioisotope may generate gamma rays, beta particles, or positrons which are converted into gamma rays upon collision with electrons.
  • the radioisotope may be used in diagnostic imaging procedures including gamma scintigraphy, specific photon emission computerized tomography, or positron emission tomography, or may be used therapeutically.
  • the addition of ascorbic acid to achieve concentrations between about 0.001 wt% and 5 wt% to the composition post-labeling increases the stability of the radiolabeled composition, with apparent increased resistance to oxidation and/or radiolytic decomposition. If excess stannous or stannic ions are present in the composition of the labeled peptide or protein, the addition of ascorbic acid also prevents formation of Sn-colloids and other radiochemical impurities.
  • compositions in which ascorbic acid has been present prior to completion of radiolabeling
  • the stabilizing agent ascorbic acid or a derivative thereof
  • the stabilizing agent be added to the composition containing a radiolabeled peptide or protein only after radiolabeling is complete (i.e. , after the label has been attached to the peptide or protein and any incubation period has elapsed).
  • antioxidants notably ascorbic acid (and derivatives thereof)
  • antioxidants inhibit radiolysis and free radical damage to proteins or peptides labeled with isotopes emitting high energy photons or particles, such as the 2 MeV beta particle emitted upon decay of 188 Re.
  • Free radical degradation in particular can be significant, and generally increases with both time and incre sed amounts of radioactivity in the preparation.
  • ascorbic acid or derivative has the added advantage in a pharmaceutical or clinical setting for the purpose of "bulking" the preparation, that is, giving the preparation a certain required minimum volume for easy manipulation and use.
  • preparations of ascorbic acid (or derivative) for injection are commercially available, and exhibit little or no toxicity. If not available, they can be synthesized using known techniques.
  • a protein or peptide radiolabeling kit may be employed, which will contain one or more vials containing the protein or peptide to be radiolabeled, and may contain a transfer ligand, a reducing agent, depending on the radioisotope to be employed, other radiolabeling reagents and one or more excipients.
  • the contents of a vial may be lyophilized, frozen or in liquid formulation.
  • the radioisotope typically in an aqueous solution, is added to the vial containing the protein or peptide to be radiolabeled to initiate radiolabeling. It is key that the radiolabeling vial not include ascorbic acid or derivative since their presence may adversely affect radiolabeling.
  • ascorbic acid and other stabilizing agents are conventionally included in the radiolabeling kit in a separate vial, or may be obtained from other sources in a separate vial.
  • the radiolabeling vial is allowed to incubate for a period of time, generally ranging from 15 minutes to one hour, at a temperature ranging from room temperature to 100°C. After the incubation period is completed, unreacted label and/or excess reducing agent are optionally removed, the ascorbic acid or derivative thereof then added to the radiolabeled protein or peptide in the radiolabeling vial.
  • compositions containing ascorbic acid may be used, typically containing 500 mg/2 ml ascorbic acid, with the volume added ranging from 10 ⁇ l to 2 ml, and preferably from 250 ⁇ to 1 ml.
  • the derivative may be a physiologically acceptable water soluble salt of ascorbic acid, such as sodium ascorbate, potassium ascorbate, lithium ascorbate, etc. or ester of ascorbic acid.
  • an equivalent amount is used.
  • the composition may be prepared for administration.
  • the radiolabeled composition is administered parenterally, and most commonly intravenously, but other forms of administration are contemplated and possible.
  • the stabilizer of the invention is added as soon as radiolabeling is complete. If any purification of the radiolabeled protein is necessary, the stabilizer can be added after such purification. It is an advantage of the invention, however, that incorporation of the stabilizer in compositions containing excess stannous ion prevents formation of quantities of stannous colloids and other radiochemical impurities that would interfere with performance of the radiolabeled protein or peptide for its intended imaging diagnostic or therapeutic purpose.
  • RC-160 is a cyclic somatostatin analogue with the general structure:
  • Radiolabeling kits were prepared using aseptic techniques, with each kit prepared in a 10 ml serum vial using a 2 ml liquid fill.
  • the liquid fill contained 200 ⁇ g of RC-160 peptide in 45 mM sodium potassium tartrate, 10 mM potassium hydrogen phthalate buffer, pH 5.0, in 5 mM stannous tartrate with 1 % maltose added as a freeze-drying excipient.
  • Each kit contained a maximum of 1 ⁇ 9 ⁇ g of tin. After filling, the vials were lyophilized, the head space gas filled with nitrogen, and the vials stoppered and crimped. Lyophilized vials were then stored refrigerated at 2-8°C.
  • a kit 4-5 ml of 188 Re-perrhenate solution containing 10-100 mCi was added to the kits, and the kits then heated in a boiling water bath for 30-45 minutes. Following a brief cooling period, 2 ml of ascorbic acid for Injection, U.S.P., was added to the labeled kit through a 0.22 micron filter. Two types of parenteral ascorbate were used with similar results, Ascorbic Acid for Injection, U.S.P., 500 mg/2 ml, and AscorvitTM (a preparation of ascorbate) 100 mg (Jenapharm, Germany). An elution profile from an analytical HPLC at 6-hours post-labeling is shown in Fig. 1.
  • RC-160 was radiolabeled at pH 5.0 for 30 minutes at 90°C and ascorbate was added post- labeling.
  • the temperature effect is shown in Fig. 2: 19 minutes for 37°C and 21 minutes for 90°C.
  • Fig. 1 is an elution profile of Re-188-RC
  • RP-HPLC revealed an elution profile indicative of inefficient radiolabeling in the presence of this low amount of ascorbic acid.
  • the RP-HPLC results were confirmed by TLC.
  • the radiolabeling kit could be formulated with an excess of stannous ions and RC-160 to accommodate a variety of labeling situations. In the presence of .
  • 188 Re the RC-160 and stannous ions interact to result in what is believed to be metal-cyclized 188 Re-RC-160.
  • T he 18 *Re-RC-160 was demonstrated by RP-HPLC not to be identical with stannous-ion-reduced RC-160, or RC-160 reduced with dithiothreitol. Since 188 Re is produced essentially carrier-free from a 188W/ 188 Re generator, it is hypothesized that excess stannous ions will reduce the RC-160 not complexed to 188 Re.
  • each vial was radiolabeled by the addition of "Tc sodium pertechnetate in a volume of 1.0 ml saline, followed by incubation for 30 minutes at 37°C. The radiolabeled antibody was then evaluated.
  • each vial was radiolabeled by the addition of
  • the net immunoreactive fraction was 51 % when ascorbic acid stabilizing agent was added, and 46% when it was not added. In another lot of kits, the net immunoreactive fraction was 59% when ascorbic acid stabilizing agent was added, and 42% when it was not added.
  • a radiolabeling kit was formulated which included a 3 ml reaction vial containing the following lyophilized components:
  • the formulation pH is adjusted to 6.2 ⁇ 0.1 with NaOH and/or HCl prior to dispensing 0.50 ml per vial and lyophilizing.
  • the product contains no bacteriostatic preservative.
  • Minimum stannous tin is 10 micrograms and total tin is 24 micrograms.
  • One 2 ml ampule of commercially available Ascorbic Acid Inject ion, U.S. P. (Ascorbic Acid 250 mg/ml and Edetate Disodium 0.025 % in water for injection) is included in the radiolabeling kit.
  • Ascorbic Acid solution (0.75 ml) is added after ""Tc labeling as a stabilizer for the reconstituted product.
  • the 3 ml reaction vial with lyophilized components is removed from 2 -8°C storage and allowed to come to room temperature. Without the addition of air, the contents of the vial are aseptically reconstituted with up to 40 mCi (1500 MBq) of Sodium Pertechnetate ""Tc Injection (0.25 ml) from a fresh generator elution. The vial is gently swirled until the lyophilized product is completely dissolved. The reaction vial containing Sodium Pertechnetate ""Tc Injection is incubated for 30 minutes at 37°C.
  • kits were prepared as in Example 3, except that one kit contained a total of 18 g of stannous tartrate, another kit contained a total of 24 g of stannous tartrate, and a third kit contained a total 30 g of stannous tartrate.
  • Each kit was labeled with 20 mCi of ""Tc sodium pertechnetate in a volume of 250 ⁇ l, with incubation for 30 minutes at 37°C. Following incubation, 50 ⁇ l of each radiolabeled preparation was removed, to which was added 150 ⁇ l of an ascorbic acid solution. Each of the resulting six different preparations was tested for immunoreactivity as described in Example 2.
  • kits were formulated using polyclonal human gamma globulin as the protein to be radiolabeled.
  • a variety of reducing agents including dithionite, sulfite, and tetrathionate were employed, wit h stannous used to reduce the ""Tc sodium pertechnetate.
  • Each type of different kit was radiolabeled both with and without ascorbic acid in the radiolabeling solution, and in each case the presence of ascorbic acid during radiolabeling adversely affected labeling yields, blood levels obtained upon intravenous injection of the radiolabeled preparation into experimental animals, and the resistance of the labeled antibody to challenge with cysteine.
  • FIG 10 shows HPLC profiles of ""Tc- labeled IgM samples, showing the difference in yield over time with labeling occurring in the presence of ascorbate on the one hand, and addition of ascorbate post-labeling on the other.
  • the top panel illustrates results obtained with ascorbate (50 mg/ml, pH 6.G), sodium sulfite (1 mg/ml, pH 7.4) or sodium bisulfite (1 mg/ml, pH 5.5).
  • the bottom panel illustrates results obtained with Ascorbate for Injection, USP, or the Ascorvit formulation.

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Abstract

Utilisation d'acide ascorbique et de ses dérivés pour stabiliser des protéines et des peptides radiomarqués en évitant la perte du radio marquage par oxydation et l'autoradiolyse. L'acide ascorbique est ajouté après le radiomarquage, avec éventuellement la période d'incubation requise, mais avant l'administration au patient.
PCT/US1997/001695 1996-02-02 1997-02-03 Stabilisation apres marquage de proteines et de peptides radiomarquees WO1997028181A2 (fr)

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CA002279349A CA2279349C (fr) 1996-02-02 1997-02-03 Composition pharmaceutique stabilisee avec un ascorbate et methode pour son utilisation
AU25264/97A AU2526497A (en) 1996-02-02 1997-02-03 Post-labeling stabilization of radiolabeled proteins and peptides

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US1102796P 1996-02-02 1996-02-02
US60/011,027 1996-05-21

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Cited By (20)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002004030A3 (fr) * 2000-07-06 2003-02-27 Bristol Myers Squibb Pharma Co Compositions radiopharmaceutiques stables et procedes de fabrication de ces compositions
WO2003000271A3 (fr) * 2001-06-22 2003-04-17 Univ Alberta The University Of Systeme et procede d'etiquetage a grande echelle de composes a radiohalogenes
EP1365813A4 (fr) * 2001-02-26 2006-03-22 Bristol Myers Squibb Pharma Co Analogues a l'acide ascorbique pour produits metalloradiopharmaceutiques
US7258869B1 (en) 1999-02-08 2007-08-21 Alza Corporation Stable non-aqueous single phase viscous vehicles and formulations utilizing such vehicle
US7919109B2 (en) 1999-02-08 2011-04-05 Intarcia Therapeutics, Inc. Stable non-aqueous single phase viscous vehicles and formulations utilizing such vehicles
JP2012516328A (ja) * 2009-01-30 2012-07-19 ネオプローブ コーポレイション ジエチレントリアミン五酢酸（ｄｔｐａ）−デキストランを放射標識するための組成物
US9572889B2 (en) 2008-02-13 2017-02-21 Intarcia Therapeutics, Inc. Devices, formulations, and methods for delivery of multiple beneficial agents
US9682127B2 (en) 2005-02-03 2017-06-20 Intarcia Therapeutics, Inc. Osmotic delivery device comprising an insulinotropic peptide and uses thereof
US9724293B2 (en) 2003-11-17 2017-08-08 Intarcia Therapeutics, Inc. Methods of manufacturing viscous liquid pharmaceutical formulations
WO2017155937A1 (fr) 2016-03-07 2017-09-14 Actinium Pharmaceuticals, Inc. Compositions d'immunoglobulines anti-cd45 radio-marquées stabilisées
US9889085B1 (en) 2014-09-30 2018-02-13 Intarcia Therapeutics, Inc. Therapeutic methods for the treatment of diabetes and related conditions for patients with high baseline HbA1c
USD835783S1 (en) 2016-06-02 2018-12-11 Intarcia Therapeutics, Inc. Implant placement guide
US10159714B2 (en) 2011-02-16 2018-12-25 Intarcia Therapeutics, Inc. Compositions, devices and methods of use thereof for the treatment of cancers
US10231923B2 (en) 2009-09-28 2019-03-19 Intarcia Therapeutics, Inc. Rapid establishment and/or termination of substantial steady-state drug delivery
USD860451S1 (en) 2016-06-02 2019-09-17 Intarcia Therapeutics, Inc. Implant removal tool
US10501517B2 (en) 2016-05-16 2019-12-10 Intarcia Therapeutics, Inc. Glucagon-receptor selective polypeptides and methods of use thereof
US10527170B2 (en) 2006-08-09 2020-01-07 Intarcia Therapeutics, Inc. Osmotic delivery systems and piston assemblies for use therein
US10835580B2 (en) 2017-01-03 2020-11-17 Intarcia Therapeutics, Inc. Methods comprising continuous administration of a GLP-1 receptor agonist and co-administration of a drug
US10925639B2 (en) 2015-06-03 2021-02-23 Intarcia Therapeutics, Inc. Implant placement and removal systems
US11246913B2 (en) 2005-02-03 2022-02-15 Intarcia Therapeutics, Inc. Suspension formulation comprising an insulinotropic peptide

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ATE481963T1 (de) 2006-05-30 2010-10-15 Intarcia Therapeutics Inc Zweiteiliger flussmodulator mit einem internen kanal für ein osmotisches ausgabesystem

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4401646A (en) * 1981-05-08 1983-08-30 University Patents Inc. Method and apparatus for purifying materials radiolabeled with technetium-99m
US4988496A (en) * 1988-05-31 1991-01-29 Neorx Corporation Metal radionuclide chelating compounds for improved chelation kinetics
US5011676A (en) * 1990-03-27 1991-04-30 Thomas Jefferson University Method to directly radiolabel antibodies for diagnostic imaging and therapy
DK0600992T3 (da) * 1991-08-29 2000-10-09 Mallinckrodt Medical Inc Anvendelse af gentissyre eller gentisylalkohol til stabilisering af radiomærkede peptider og proteiner
US5225180A (en) * 1991-09-10 1993-07-06 Diatech, Inc. Technetium-99m labeled somatostatin-derived peptides for imaging

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US7258869B1 (en) 1999-02-08 2007-08-21 Alza Corporation Stable non-aqueous single phase viscous vehicles and formulations utilizing such vehicle
US7919109B2 (en) 1999-02-08 2011-04-05 Intarcia Therapeutics, Inc. Stable non-aqueous single phase viscous vehicles and formulations utilizing such vehicles
US8048438B2 (en) 1999-02-08 2011-11-01 Intarcia Therapeutics, Inc. Stable non- aqueous single phase viscous vehicles and formulations utilizing such vehicles
WO2002004030A3 (fr) * 2000-07-06 2003-02-27 Bristol Myers Squibb Pharma Co Compositions radiopharmaceutiques stables et procedes de fabrication de ces compositions
EP1365813A4 (fr) * 2001-02-26 2006-03-22 Bristol Myers Squibb Pharma Co Analogues a l'acide ascorbique pour produits metalloradiopharmaceutiques
WO2003000271A3 (fr) * 2001-06-22 2003-04-17 Univ Alberta The University Of Systeme et procede d'etiquetage a grande echelle de composes a radiohalogenes
US7238338B1 (en) 2001-06-22 2007-07-03 Triumf System and method for the large scale labeling of compounds with radiohalogens
US9724293B2 (en) 2003-11-17 2017-08-08 Intarcia Therapeutics, Inc. Methods of manufacturing viscous liquid pharmaceutical formulations
US9682127B2 (en) 2005-02-03 2017-06-20 Intarcia Therapeutics, Inc. Osmotic delivery device comprising an insulinotropic peptide and uses thereof
US11246913B2 (en) 2005-02-03 2022-02-15 Intarcia Therapeutics, Inc. Suspension formulation comprising an insulinotropic peptide
US10363287B2 (en) 2005-02-03 2019-07-30 Intarcia Therapeutics, Inc. Method of manufacturing an osmotic delivery device
US10527170B2 (en) 2006-08-09 2020-01-07 Intarcia Therapeutics, Inc. Osmotic delivery systems and piston assemblies for use therein
US9572889B2 (en) 2008-02-13 2017-02-21 Intarcia Therapeutics, Inc. Devices, formulations, and methods for delivery of multiple beneficial agents
US10441528B2 (en) 2008-02-13 2019-10-15 Intarcia Therapeutics, Inc. Devices, formulations, and methods for delivery of multiple beneficial agents
JP2015164933A (ja) * 2009-01-30 2015-09-17 ナビディア、バイオファーマスーティカルズ、インコーポレイテッドＮａｖｉｄｅａ Ｂｉｏｐｈａｒｍａｃｅｕｔｉｃａｌｓ， Ｉｎｃ． ジエチレントリアミン五酢酸（ｄｔｐａ）−デキストランを放射標識するための組成物
JP2017066148A (ja) * 2009-01-30 2017-04-06 ナビディア、バイオファーマスーティカルズ、インコーポレイテッドＮａｖｉｄｅａ Ｂｉｏｐｈａｒｍａｃｅｕｔｉｃａｌｓ， Ｉｎｃ． ジエチレントリアミン五酢酸（ｄｔｐａ）−デキストランを放射標識するための組成物
JP2012516328A (ja) * 2009-01-30 2012-07-19 ネオプローブ コーポレイション ジエチレントリアミン五酢酸（ｄｔｐａ）−デキストランを放射標識するための組成物
US10231923B2 (en) 2009-09-28 2019-03-19 Intarcia Therapeutics, Inc. Rapid establishment and/or termination of substantial steady-state drug delivery
US10869830B2 (en) 2009-09-28 2020-12-22 Intarcia Therapeutics, Inc. Rapid establishment and/or termination of substantial steady-state drug delivery
US12042557B2 (en) 2009-09-28 2024-07-23 I2O Therapeutics, Inc. Rapid establishment and/or termination of substantial steady-state drug delivery
US10159714B2 (en) 2011-02-16 2018-12-25 Intarcia Therapeutics, Inc. Compositions, devices and methods of use thereof for the treatment of cancers
US9889085B1 (en) 2014-09-30 2018-02-13 Intarcia Therapeutics, Inc. Therapeutic methods for the treatment of diabetes and related conditions for patients with high baseline HbA1c
US10583080B2 (en) 2014-09-30 2020-03-10 Intarcia Therapeutics, Inc. Therapeutic methods for the treatment of diabetes and related conditions for patients with high baseline HbA1c
US10925639B2 (en) 2015-06-03 2021-02-23 Intarcia Therapeutics, Inc. Implant placement and removal systems
WO2017155937A1 (fr) 2016-03-07 2017-09-14 Actinium Pharmaceuticals, Inc. Compositions d'immunoglobulines anti-cd45 radio-marquées stabilisées
EP3426305A4 (fr) * 2016-03-07 2020-01-08 Actinium Pharmaceuticals, Inc. Compositions d'immunoglobulines anti-cd45 radio-marquées stabilisées
US11214607B2 (en) 2016-05-16 2022-01-04 Intarcia Therapeutics Inc. Glucagon-receptor selective polypeptides and methods of use thereof
US10501517B2 (en) 2016-05-16 2019-12-10 Intarcia Therapeutics, Inc. Glucagon-receptor selective polypeptides and methods of use thereof
US11840559B2 (en) 2016-05-16 2023-12-12 I2O Therapeutics, Inc. Glucagon-receptor selective polypeptides and methods of use thereof
USD912249S1 (en) 2016-06-02 2021-03-02 Intarcia Therapeutics, Inc. Implant removal tool
USD840030S1 (en) 2016-06-02 2019-02-05 Intarcia Therapeutics, Inc. Implant placement guide
USD835783S1 (en) 2016-06-02 2018-12-11 Intarcia Therapeutics, Inc. Implant placement guide
USD962433S1 (en) 2016-06-02 2022-08-30 Intarcia Therapeutics, Inc. Implant placement guide
USD860451S1 (en) 2016-06-02 2019-09-17 Intarcia Therapeutics, Inc. Implant removal tool
US10835580B2 (en) 2017-01-03 2020-11-17 Intarcia Therapeutics, Inc. Methods comprising continuous administration of a GLP-1 receptor agonist and co-administration of a drug
US11654183B2 (en) 2017-01-03 2023-05-23 Intarcia Therapeutics, Inc. Methods comprising continuous administration of exenatide and co-administration of a drug

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CA2279349A1 (fr) 1997-08-07
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WO1997028181A3 (fr) 1997-10-09

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