Claims for Patent: 5,482,698

Summary for Patent: 5,482,698

Title:	Detection and therapy of lesions with biotin/avidin polymer conjugates
Abstract:	Methods of detecting and/or treating lesions in a patient are provided. The methods are an improvement over known methods comprising the steps of (a) parenterally injecting a subject with a targeting composition comprised of a biotin-protein conjugate or an avidin-protein conjugate, wherein the protein preferentially binds to a marker substance produced or associated with the targeted lesion, and allowing the protein conjugate to preferentially accrete at the targeted lesion; (b) then parenterally injecting a clearing composition comprised of (i) avidin, when the targeting composition is a biotin-protein conjugate, or (ii) biotin, when the targeting composition is a avidin-protein conjugate, and allowing the clearing composition to substantially clear the targeting composition from non-targeted sites and to bind to the targeting composition accreted at the targeted lesion; and (c) parenterally injecting a detection or therapeutic composition comprised of a conjugate of (i) avidin and detection or therapeutic agent when the clearing composition is biotin, or (ii) biotin and detection or therapeutic agent when the clearing agent is avidin, and allowing the composition to accrete at the targeted lesion. The improvement is having at least one of the compositions of step (a) or (b) further comprise a polymer to which multiple moieties of avidin or biotin can conjugate, thereby providing an increased number of binding sites to which a subsequently administrated composition can bind thereby amplifying the amount of detection or therapeutic agent at the targeted site.
Inventor(s):	Griffiths; Gary L. (Morristown, NJ)
Assignee:	Immunomedics, Inc. (Morris Plains, NJ)
Application Number:	08/051,144
Patent Claims:	1. An improvement in a method of detecting or treating lesions in a subject, the method comprising the steps of (a) parenterally injecting a subject with a targeting composition comprised of (i) a biotin-protein conjugate or (ii) an avidin-protein conjugate, wherein the protein of said conjugate binds to a marker substance produced by or associated with the targeted lesion, and allowing said conjugate to accrete at the targeted lesion; (b) then parenterally injecting a clearing composition comprised of (i) avidin, when said targeting composition is a biotin-protein conjugate, or (ii) biotin, when said targeting composition is an avidin-protein conjugate, and allowing said clearing composition to substantially clear said targeting composition from non-targeted sites and to bind to said targeting composition accreted at the targeted lesion; and (c) parenterally injecting a detection or therapeutic composition comprised of a conjugate of (i) avidin and a detection or therapeutic agent when said clearing composition is biotin, or (ii) biotin and a detection or therapeutic agent when said clearing agent is avidin, and allowing said detection or therapeutic composition to accrete at the targeted lesion thereby treating the lesion when said agent is a therapeutic agent, or if said agent is a detection agent, (d) detecting said agent accreted at the lesion, thereby detecting the lesion; wherein the improvement is that at least one of the compositions of step (a) or (b) further comprises a polymer comprising multiple avidin or biotin binding sites, thereby providing an increased number of binding sites to which a subsequently administered composition of step (b) or (c) binds, thereby amplifying the amount of said detection or therapeutic agent at the targeted lesion. 2. A method of detecting or treating lesions in a subject, the method comprising the steps of (a) injecting the subject with a first composition comprised of (1) a multibiotin-polymer-protein conjugate, or (2) a biotin-protein conjugate, wherein the protein of said conjugate binds to a marker substance produced by or associated with the target lesion, and allowing said conjugate to accumulate at the target lesion to provide biotin molecules at the lesion; (b) injecting a clearing agent comprised of (1) avidin, or (2) a multiavidin-polymer conjugate, or (3) a multiavidin-multitherapeutic agent-polymer conjugate or a multiavidin-multidetection agent-polymer conjugate, or (4) an avidin-therapeutic agent conjugate or an avidin-detection agent conjugate, or (5) a multiavidin-therapeutic agent-polymer conjugate or a multiavidin-detection agent-polymer conjugate, and permitting said clearing agent to remove circulating biotin conjugate; (c) injecting a localizing agent comprised of (1) avidin, or (2) a multiavidin-polymer conjugate, or (3) a multiavidin-multitherapeutic agent-polymer conjugate or a multiavidin-multidetection agent-polymer conjugate, or (4) an avidin-therapeutic agent conjugate or an avidin-detection agent conjugate, or (5) a multiavidin-therapeutic agent-polymer conjugate or a multiavidin-detection agent-polymer conjugate, wherein said localizing agent is the same as or different from said clearing agent and is injected concurrently with or subsequent to said clearing agent, and permitting said localizing agent to localize at the target lesion by binding to a biotin molecule; (d) injecting a composition comprising (1) a biotin-therapeutic agent conjugate or a biotin-detection agent conjugate, or (2) a polymer-multibiotin conjugate or, (3) a polymer-multibiotin-multi-therapeutic agent conjugate or a polymer-multibiotin-multi-detection agent conjugate, and permitting biotin molecules of said composition of step (d) to bind to avidin molecules of said localizing agent of step (c); (e) if a conjugate containing polymer is used in step (d), then injecting a composition comprising (1) an avidin-therapeutic agent conjugate or an avidin-detection agent conjugate, or (2) a multiavidin-polymer conjugate, or (3) avidin, or (4) a multiavidin-multitherapeutic agent-polymer conjugate or a multiavidin-multidetection agent-polymer conjugate, and permitting an avidin molecule of said composition of step (e) to bind to a biotin molecule of said localizing composition of step (d); (f) if other than said avidin-therapeutic agent conjugate or said avidin-detection agent conjugate has been injected in step (e), injecting a biotin-therapeutic agent conjugate or a biotin-detection agent conjugate and permitting a biotin molecule of said conjugate of step (f) to bind to an avidin molecule at the target lesion thereby treating the lesion when said agent is a therapeutic agent, or if said agent is a detection agent, (g) detecting said agent bound at the lesion, thereby detecting the targeted lesion; wherein a polymer conjugate is used in at least one step and the polymer has a sufficiently rigid stereochemical structure so that only one avidin or biotin molecule of each conjugate binds to a molecule already present at the target lesion. 3. A method of detecting or treating lesions in a subject comprising the steps of (a) injecting a subject with a first composition comprised of a multiavidin-polymer-protein conjugate or an avidin-protein conjugate, wherein the protein of said conjugate binds to a marker substance produced by or associated with the target lesion, and allowing said conjugate to accrete at the target lesion; (b) injecting a clearing agent comprised of (1) biotin, or (2) a multibiotin-polymer conjugate, or (3) a multibiotin-multitherapeutic agent-polymer conjugate or a multibiotin-multidetection agent-polymer conjugate, or (4) a biotin-therapeutic agent conjugate or a biotin-detection agent conjugate, and permitting said clearing agent to remove circulating conjugate of step (a); (c) injecting a localizing agent comprised of (1) a multibiotin-polymer conjugate, or (2) a multibiotin-multitherapeutic agent-polymer conjugate or a multibiotin-multidetection agent-polymer conjugate, or (3) a biotin-therapeutic agent conjugate or a biotin-detection agent conjugate, wherein said localizing agent is the same as or different from said clearing agent and is injected concurrently with or subsequent to said clearing agent and permitting said localizing agent to localize at the target lesion by binding to an avidin molecule; (d) injecting a composition comprised of (1) an avidin-therapeutic agent conjugate or an avidin-detection agent conjugate, or (2) a polymer-multiavidin conjugate or, (3) a polymer-multiavidin-multitherapeutic agent conjugate or a polymer-multiavidin-multidetection agent conjugate, and permitting avidin molecules of said conjugate of step (d) to bind to biotin molecules of said localizing agent; (e) if a conjugate containing polymer is used in step (d), then injecting a composition consisting essentially of (1) a biotin-therapeutic agent conjugate or a biotin-detection agent conjugate, or (2) a multibiotin-polymer conjugate, or (3) a multibiotin-multitherapeutic agent-polymer conjugate or a multibiotin-multidetection agent-polymer conjugate, and permitting the biotin molecules of the conjugate of step (e) to bind to avidin molecules provided by said composition of step (d); (f) if other than a biotin-therapeutic agent or a biotin-detection agent has been injected in step (e), injecting an avidin-therapeutic agent conjugate or an avidin-detection agent conjugate and permitting the avidin molecules of the conjugate of step (f) to bind to biotin molecules at the target lesion thereby treating the lesion when said agent is a therapeutic agent, or if said agent is a detection agent; (g) detecting said agent bound at the lesion, thereby detecting the targeted lesion; wherein a polymer conjugate is used in at least one step of the method and the polymer has a sufficiently rigid stereochemical structure so that only one biotin or avidin molecule of each conjugate binds to a molecule already present at the target lesion. 4. The method of claim 2 wherein: said composition of step (a) comprises a multibiotin-polymer-protein conjugate; and provides multiple biotin molecules at the targeted lesion; said clearing agent of step (b) and said localizing agent of step (c) are comprised of avidin; and said composition of step (d) comprises a biotin-detection agent conjugate or a biotin-therapeutic agent conjugate. 5. The method of claim 2 wherein: said composition of step (a) is comprised of a biotin-protein conjugate, and provides biotin molecules at the target lesion; said clearing agent of step (b) and said localizing agent of step (c) are comprised of a multiavidin-polymer conjugate, wherein only one avidin molecule of each multiavidin-polymer conjugate binds to a biotin molecule at the target lesion; and said composition of step (d) comprises a biotin-therapeutic agent conjugate or a biotin-detection agent conjugate. 6. The method of claim 2 wherein: said composition of step (a) is comprised of a biotin-protein conjugate and provides biotin molecules at the target lesion; said clearing agent of step (b) and said localizing agent of step (c) are comprised of a multiavidin-polymer-therapeutic agent conjugate or a multiavidin-polymer-detection agent conjugate, wherein only one avidin molecule of each conjugate binds to a biotin molecule present at the target lesion, and said composition of step (d) comprises a biotin-therapeutic agent conjugate or a biotin-detection agent conjugate. 7. The method of claim 3 wherein: said composition of step (a) is comprised of an avidin-protein conjugate and provides avidin molecules at the target lesion; said clearing agent of step (b) and said localizing agent of step (c) are comprised of a multibiotin-polymer conjugate, wherein only one biotin molecule of each multibiotin-polymer conjugate binds to an avidin molecule provided by each avidin-protein conjugate accreted at the target lesion, whereby each multibiotin-polymer conjugate provides multiple biotin molecules at the target lesion; and said composition of step (d) comprises an avidin-therapeutic agent conjugate or an avidin-detection agent conjugate. 8. The method of claim 3 wherein: said composition of step (a) is comprised of an avidin-protein conjugate and provides avidin molecules at the target lesion; wherein said clearing agent of step (b) and said localizing agent of step (c) are comprised of a multibiotin-polymer-therapeutic agent conjugate or a multibiotin-polymer-detection agent conjugate, wherein only one biotin molecule of each multibiotin-polymer-therapeutic agent conjugate or multibiotin-polymer-detection agent conjugate binds to an avidin molecule provided by each avidin-protein conjugate of step (a) accreted at the target lesion, whereby each multibiotin-polymer-therapeutic agent conjugate or multibiotin-polymer-detection agent conjugate provides multiple biotin molecules and provides a therapeutic agent or detection agent to the target lesion; and said composition of step (d) comprises an avidin-therapeutic agent conjugate or an avidin-detection agent conjugate. 9. The method of claim 1, wherein the polymer is selected from the group consisting of polyamino-acids, oligonucleotides, polyimines, polysaccharides, polyamines, polycarboxylic acids and polyalcohols. 10. The method of claim 9 wherein the polymer has restricted stereochemistry such that each of the multiple biotin or avidin moieties conjugated to said polymer binds a complementary avidin or biotin moiety subsequently administered, and only one of the said multiple biotin or avidin moieties conjugated to said polymer binds to a complementary avidin or biotin moiety already present at the target site. 11. The method of claim 9, wherein the polymer has a molecular weight of 5,000 to 200,000 kD. 12. The method of claim 1 wherein the polymer is a dextran or a starburst dendrimer. 13. The method of claim 12 wherein the polymer is an aminodextran. 14. The method of claim 1, wherein the lesion is cancerous, cardiovascular, infectious or inflammatory. 15. The method of claim 14, wherein the cardiovascular lesion is a thrombus, embolus, infarct or atherosclerotic plaque. 16. The method of claim 14, wherein the cancerous lesion is a carcinoma, melanoma, sarcoma, neuroblastoma, leukemia, lymphoma, glioma or myeloma. 17. The method of claim 14, wherein the lesion is infectious or inflammatory. 18. The method of claim 1, wherein the protein is selected from the group consisting of a peptide, polypeptide, hormone, lymphokine, growth factor, albumin, cytokine, enzyme, immune modulator, receptor protein, antibody and antibody fragment. 19. The method of claim 18, wherein the protein is one of monoclonal antibody, or a specific binding fragment thereof. 20. The method of claim 19, wherein the fragment is one of a Fv, single chain antibody, Fab, Fab', F(ab).sub.2 or F(ab').sub.2. 21. The method of claim 20, wherein the fragment is one of Fab, Fab', F(ab).sub.2 or F(ab').sub.2. 22. The method of claim 19, wherein the antibody is multispecific. 23. The method of claim 22, wherein the antibody is multispecific to differing epitopes or molecules of a marker substance. 24. The method of claim 18, wherein the protein has a specific immunoreactivity to a marker substance of at least 60% and a cross-reactivity to other antigens or non-target substances of less than 35%. 25. The method of claim 1, wherein the method is for detection of a lesion. 26. The method of claim 25, wherein the method comprises one of external imaging of said patient or internal detection of said patient. 27. The method of claim 26, wherein said internal detection is effected during an operative, intravascular or endoscopic procedure. 28. The method of claim 25, wherein the detection agent is one of a radionuclide, mri enhancing agent, photoactivated dye or differentiation agent. 29. The method of claim 25, wherein the detection agent is one of a gamma-emitter, positron-emitter, x-ray-emitter, beta-emitter or fluorescence-emitter. 30. The method of claim 28, wherein the detection agent is a radionuclide with an energy between 10 and 7000 keV. 31. The method of claim 29, wherein the radionuclide used for external imaging is one of Iodine-123, Iodine-131, Indium-111, Gallium-67, Ruthenium-97, Technetium-99m, Cobalt-57, Cobalt-58, Chromium-51, Iron-59, Selenium-75, Thallium-201, or Ytterbium-169. 32. The method of claim 29, wherein the radionuclide used for internal detection is Iodine-125, Iodine-123, Iodine-131, Indium-111, Technetium-99m or Gallium-67. 33. The method of claim 28, wherein the mri enhancing agent is a species of one of Gadolinium, Iron, Manganese, Rhenium, Europium, Lanthanium, Holmium, or Terbium. 34. The method of claim 1, wherein the method is for treating a lesion. 35. The method of claim 34, wherein the therapeutic agent is one of an isotope, drug, toxin, fluorescent dye activated by nonionizing radiation, hormone, hormone antagonist, receptor antagonist, enzyme or proenzyme. 36. The method of claim 35, wherein the therapeutic agent is an electron- or neutron-capturing agent. 37. The method of claim 35, wherein the therapeutic agent is an isotope which is one of Iodine-125, Iodine-131, Actinium-225, Bismuth-212, Bismuth-213, Lead-212, Rhenium-186, Rhenium-188, Silver-111, Platinum-197, Palladium-109, Copper-67, Phosphorus-32, Phosphorus-33, Yttrium-90, Scandium-47, Samarium-153, Lutetium-177, Rhodium-105, Praseodymium-142, Praseodymium-143, Terbium-161, Holmium-166, or Gold-199. 38. The method of claim 34, wherein the therapeutic agent is an anti-DNA, anti-RNA, radiolabeled oligonucleotide, anti-protein or anti-chromatin cytoxic or antimicrobial agent. 39. The method of claim 36, wherein the therapeutic agent is a drug which is one of taxol, mechlorethamine, cyclophosphamide, melphalan, uracil mustard, chlorambucil, thiotepa, busulfan, carmustine, lomustine, semustine, streptozocin, dacarbazine, methotrexate, fluorouracil, cytarabine, azaribine, mercaptopurine, thioguanine, vinblastine, vincristine, dactinomycin, daunorubicin, doxorubicin, bleomycin, mithramycin, mitomycin, L-asparaginase, cisplatin, hydroxyurea, procarbazine, mitotane, prednisone, hydroxyprogesterone caproate, medroprogesterone acetate, diethylstilbestrol, ethinyl estradiol, tamoxifen, and testosterone propionate or fluoxymesterone. 40. The method of claim 35, wherein the therapeutic agent is a toxin which is one of abrin, alpha toxin, diphtheria toxin, exotoxin, gelonin, pokeweed antiviral protein, ricin, or saporin. 41. A sterile injectable composition for human use comprising a detection or therapeutic composition comprised of (i) a multiavidin-polymer-protein conjugate, (ii) a multiavidin-polymer conjugate, (iii) a multiavidin-polymer-detection agent conjugate or multiavidin-polymer-therapeutic agent conjugate, (iv) a multibiotin-polymer conjugate (v) a multibiotin-polymer-protein conjugate or (vi) a multibiotin-polymer-therapeutic agent conjugate or a multibiotin-polymer detection agent conjugate, wherein the protein of said conjugate preferentially binds to a marker substance produced by or associated with a targeted lesion and wherein the polymer of said conjugate has a sufficiently rigid stereochemical structure so that one avidin or biotin molecule of the conjugate binds to one biotin or avidin molecule already present at the target lesion. 42. The method of claim 2, wherein the clearing agent of step (b) and the localizing agent of step (c) are the same, and wherein steps (b) and (c) are effected together by one injection. 43. The method of claim 3, wherein the clearing agent of step (b) and the localizing agent of step (c) are the same, and wherein steps (b) and (c) are effected together by one injection.

Details for Patent 5,482,698

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Recordati Rare Diseases, Inc.	ELSPAR	asparaginase	For Injection	101063	01/10/1978		2039-02-26