Claims for Patent: 4,636,380

Summary for Patent: 4,636,380

Title:	Novel physiologic chemical method of labeling protein substances with the radionuclides of indium
Abstract:	A novel chemical method of labeling plasma proteins, compounds and substances containing proteins with the radionuclides of indium at physiologic pH 6-8 condition producing an efficiently labeled radioactive tracer material suitable for biomedical applications. These radiolabeled protein substances are not denatured by the labeling process but retain their native physiobiological properties. This novel labeling technique provides a simple and rapid means of labeling plasma proteins such as human serum albumin, transferrin, lactoferrin, fibrinogen, and immunoglobulin or antibodies with .sup.111 In or .sup.113m In for scintigraphic imaging which may allow visualization of thrombi, emboli, myocardial infarcts, infectious lesions, tumors or any vascular abnormalities in the body.
Inventor(s):	Wong; Dennis W. (Torrance, CA)
Assignee:
Application Number:	06/602,923
Patent Claims:	1. A method of labeling mammalian plasma proteins with the radionuclides of Indium at physiologic pH 6-8 condition which comprises the sequential steps of: a. mixing a solution of stannous salt with an aqueous solution of sodium citrate at room temperature for 1-5 minutes; b. reacting the (Sn)citrate chemical species of step (a) with a radioactive solution of InCl.sub.3 at room temperature for 1-5 minutes; c. raising the pH of the radioactive mixture of step (b) to 7.4 with a sufficient amount of dilute alkaline solution; d. heating the neutralized radioactive admixture of step (c) at 120.degree. C. for 15 minutes and allow it to cool to room temperature for 5-10 minutes; e. binding of the radionuclide to the protein ligand by adding an aqueous protein solution desired to be labeled to the neutralized radioactive admixture of step (d) and incubating the radioactive protein admixture at 37.degree. for 30 minutes. 2. A method according to claim 1, wherein said radionuclides of Indium is selected from the group consisting of .sup.111 In, .sup.113m In and .sup.114 In. 3. A method according to claim 2, wherein said radionuclide of Indium is an aqueous solution of .sup.111 InCl.sub.3, .sup.113m InCl.sub.3 or .sup.114 InCl.sub.3 providing from 0.1 mCi to 100 Ci of radioactivity. 4. A method according to claim 1, wherein said stannous salt is selected from the group consisting of stannous chloride(SnCl.sub.2), stannous fluoride (SnF.sub.2) and stannous tartrate. 5. A method according to claim 4, wherein said stannous salt is present in the amount of 0.1-100 mg dissolved in 0.05-0.1N HCl solution. 6. A method according to claim 5, wherein said stannous salt is present in the amount of 0.2-5 mg per ml of 0.05N HCl solution. 7. A method according to claim 1 wherein said sodium citrate solution is an aqueous solution of 0.5-10% trisodium citrate. 8. A method according to claim 7 wherein said sodium citrate solution is an aqueous solution of 5% trisodium citrate. 9. A method according to claim 1 wherein said dilute alkaline solution is an aqueous solution of 0.1-1.0N NaOH. 10. A method according to claim 9, wherein said dilute alkaline solution is an aqueous solution of 0.1N NaOH. 11. A method according to claim 1, wherein said plasma protein is selected from the group consisting of human serum albumin, human transferrin, human lactoferrin, human fibrinogen, human immune gamma globulin and bovine thrombin. 12. A method according to claim 11, wherein said plasma protein is present in the amount of 0.1 mg to 10 grams dissolved in aqueous medium together with any pharmaceutically acceptable preservatives or stabilizing agents. 13. A method according to claim 12, wherein said plasma protein is present in the amount of 0.1-100 mg dissolved in 1-5 ml distilled water or normal saline. 14. A method of producing a protein binding radioactive bimetallic compound of Indium having a formulus of M(Sn)C.sub.6 H.sub.5 O.sub.7 where M=In-111, In-113m or In-114 at physiologic pH 6-8 condition which comprises the sequential steps of: a. mixing a solution of stannous salt with an aqueous solution of sodium citrate at room temperature for 1-5 minutes; b. reacting the (Sn)citrate chemical species of step (a) with a radioactive solution of InCl.sub.3 at room temperature for 1-5 minutes; c. raising the pH of the radioactive mixture of step (b) to 7.4 with a sufficient amount of dilute alkaline solution; d. heating the neutralized radioactive admixture of step (c) at 120.degree. C. for 15 minutes and allow it to cool to room temperature for 5-10 minutes. 15. A method according to claim 14, wherein said radionuclides of Indium is selected from the group consisting of .sup.111 In, .sup.113m In and .sup.114 In. 16. A method according to claim 15, wherein said radionuclide of Indium is an aqueous solution of .sup.111 InCl.sub.3, .sup.113m InCl.sub.3 or .sup.114 InCl.sub.3 providing from 0.1 mCi to 100 Ci of radioactivity. 17. A method according to claim 14, wherein said stannous salt is selected from the group consisting of stannous chloride, stannous fluoride and stannous tartrate. 18. A method according to claim 17, wherein said stannous salt is present in the amount of 0.1-100 mg dissolved in 0.05-0.1N HCl solution. 19. A method according to claim 18, wherein said stannous salt is present in the amount of 0.2-5 mg per ml 0.05N HCl solution. 20. A method according to claim 14, wherein said sodium citrate solution is an aqueous solution of 0.5-10% trisodium citrate. 21. A method according to claim 20, wherein said sodium citrate solution is an aqueous solution of 5% trisodium citrate. 22. A method according to claim 14, wherein said dilute alkaline solution is an aqueous solution of 0.1-1.0N NaOH. 23. A method according to claim 22, wherein said dilute alkaline solution is an aqueous solution of 0.1N NaOH. 24. A method of labeling mammalian plasma proteins with .sup.111 InCl.sub.3 at physiologic pH 6-8 condition comprising the sequential steps of: a. mixing a solution of 0.1 to 5.0 mg of SnCl.sub.2 dissolved in 0.05N HCl with 0.4 ml of a 5% sodium citrate solution at room temperature for 1-5 minutes; b. reacting the (Sn)citrate chemical species from step (a) with a solution of .sup.111 InCl.sub.3 providing 0.1-100 mCi of radioactivity at room temperature for 1-5 minutes; c. raising the pH of the radioactive mixture of step (b) to 7.4 with a sufficient amount of 0.1N NaOH solution; d. heating the neutralized radioactive admixture of step (c) at 120.degree. C. for 15 minutes and allow it to cool to room temperature for 5-10 minutes; e. adding from 0.1 mg to 100 mg of the desired protein to be labeled in 1-5 ml diluent to the radioactive admixture of step (d) and incubating said radioactive protein admixture at 37.degree. C. for 30 minutes. 25. A method according to claim 24, wherein said plasma protein is selected from the group consisting of human serum albumin, human transferrin, human lactoferrin, human fibrinogen, human immune gamma globulin and bovine thrombin. 26. A method according to claim 24, wherein said human serum albumin is labeled with .sup.111 In. 27. A method according to claim 24, wherein said human transferrin is labeled with .sup.111 In. 28. A method according to claim 24, wherein said human lactoferrin is labeled with .sup.111 In. 29. A method according to claim 24, wherein said human fibrinogen is labeled with .sup.111 In. 30. A method according to claim 24, wherein said human immune gamma globulin is labeled with .sup.111 In. 31. A method according to claim 24, wherein said bovine thrombin is labeled with .sup.111 In. 32. A method of detecting vascular abnormalities in the body of a mammal by scintigraphic imaging techniques comprising: a. administering intravenously to said mammal from 0.1 mCi to 5 mCi of .sup.111 In-serum albumin labeled according to the method of claim 24; b. scanning said mammal with a scintillation camera or a rectilinear scanner at various time intervals from 0.5 to 24 hours and daily for up to two weeks; c. observing areas of increased radioactivity at the sites of the abnormality as seen in the scintigrams. 33. A method of detecting bone marrow abnormality in man or in animal by scintigraphic imaging techniques comprising: a. administering intravenously to said mammal from 0.1 mCi to 5 mCi of .sup.111 In-transferrin labeled according to the method of claim 24; b. scanning said mammal with a scintillation camera or a rectilinear scanner at various time intervals from 0.5 to 24 hours and daily for up to two weeks; c. observing increasing or decreasing radioactivity in the reticuloendothelial system as seen in the scintigrams. 34. A method of localizing and detecting fibrin clot deposition in thromboembolic diseases, myocardial infarction or tumors in man or in animal by scintigraphic imaging procedures comprising: a. administering intravenously to said mammal from 0.1 mCi to 5 mCi of .sup.111 In-fibrinogen labeled according to the method of claim 24; b. scanning said mammal with a scintillation camera or a rectilinear scanner at various time intervals from 0.5-24 hours and daily for up to two weeks; c. observing increasing radioactivity accumulated at the sites of these lesions as seen in the scintigrams. 35. A method of localizing and detecting infectious foci in man or in animal by scintigraphic imaging procedures comprising: a. administering intravenously to said mammal from 0.1 mCi to 5 mCi of .sup.111 In-immunoglobulin labeled according to the method of claim 24 which contains the specific antibody against the antigen or microorganism; b. scanning said mammal with a scintillation camera or a rectilinear scanner at various time intervals from 0.5-24 hours and daily for up to two weeks; c. observing increasing radioactivity accumulated at the sites of the infection as seen in the scintigrams. 36. A method of localizing and detecting benign or malignant tumors in man or in animal by scintigraphic imaging techniques comprising: a. administering intravenously to said mammal from 0.1 mCi to 5 mCi of .sup.111 In-immunoglobulin labeled according to the method of claim 24 which contains the specific antibody against the specific tumor; b. scanning said mammal with a scintillation camera or a rectilinear scanner at various time intervals from 0.5-24 hours and daily for up to two weeks; c. observing increasing radioactivity accumulated at the sites of thes tumors as seen in the scintigrams.

Details for Patent 4,636,380

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Grifols Therapeutics Llc	ALBUKED, PLASBUMIN-20, PLASBUMIN-25, PLASBUMIN-5	albumin (human)	For Injection	101138	10/21/1942		2040-01-28
Baxalta Us Inc.	BUMINATE, FLEXBUMIN	albumin (human)	Injection	101452	03/03/1954		2040-01-28
Csl Behring Ag	ALBURX	albumin (human)	Injection	102366	07/23/1976		2040-01-28
Grifols Biologicals Llc	ALBUTEIN	albumin (human)	Injection	102478	08/15/1978		2040-01-28