US4331647A — Tumor localization and therapy with labeled antib…

Claim 1

Independent Claim 82 words

A method for detecting and localizing a tumor which either produces or is associated with a cytoplasmic, intracellular or cell-surface marker substance which comprises injecting a human subject parenterally with a marker-specific fragment obtained by cleavage of an antibody specific to said marker substance and radiolabeled with a pharmacologically inert radioisotope capable of detection using a photoscanning device, and subsequently scanning with said device to detect and locate the site or sites of uptake of said labeled antibody fragment by said tumor.

Claim 2

Independent Claim 82 words

A method for detecting and localizing a tumor which either produces or is associated with a cytoplasmic, intracellular or cell-surface marker substance which comprises injecting a human subject parenterally with a marker-specific fragment obtained by cleavage of an antibody specific to said marker substance and radiolabeled with a pharmacologically inert radioisotope capable of detection using a photoscanning device, and subsequently scanning with said device to detect and locate the site or sites of uptake of said labeled antibody fragment by said tumor.

Claim 3

Dependent Claim 13 words

The method of claim 1, wherein said market substance is an oncofetal antigen.

Claim 4

Dependent Claim 13 words

The method of claim 1, wherein said market substance is a placental antigen.

Claim 5

Dependent Claim 18 words

The method of claim 1, wherein said marker substance is an oncogenic or tumor or tumor virus-associated antigen.

Claim 6

Dependent Claim 25 words

The method of claim 1, wherein said marker substance is a tissue- or organ-associated antigen, an ectopic hormone or a normal antigen or variant thereof.

Claim 7

Dependent Claim 13 words

The method of claim 1, wherein said marker substance is carcinoembryonic antigen (CEA).

Claim 8

Dependent Claim 12 words

The method of claim 1, wherein said marker substance is alphafetoprotein (AFP).

Claim 9

Dependent Claim 18 words

The method of claim 1, wherein said marker substance is human chorionic gonadotropin (HCG) or the beta-subunit thereof.

Claim 10

Dependent Claim 13 words

The method of claim 1, wherein said marker substance is colon-specific antigen-p (CSAp).

Claim 11

Dependent Claim 86 words

The method of claim 1, wherein said market substance is one of prostatic acid phosphatase, pancreatic oncofetal antigen, placental alkaline phosphatase, pregnancy beta 1 -globulin, parathormone, calcitonin, tissue polypeptide antigen, T-antigen, beta 2 -microglobulin, mammary tumor-associated glycoproteins (MTGP), galactyosyl transferase-II (GT-II), gp-52 viral-associated antigen, ovarian cystadenocarcinoma-associated antigen (CA-58, CCA, TA-4), basic fetoprotein (BFP), terminal deoxynucleotidyl transferase (TdT), cytoplasmic melanoma-associated antigens, human astrocytoma-associated antigen (HAAA), common glioma antigen (CGA), glioembryonic antigen (GEA), glial fibrillary acidic protein (GFA), common meningioma antigen (CMA), ferritin, and tumor angiogenesis factor (TAF).

Claim 12

Dependent Claim 24 words

The method of claim 1, wherein said marker-specific fragment is an Fab, Fab', and F(ab') 2 fragment obtained by cleavage of said specific antibody.

Claim 13

Dependent Claim 39 words

The method of claim 1, wherein said radioisotope is one of Iodine-131, Iodine-123, Iodine-126, Iodine-133, Bromine-77, Indium-111, Indium-113m, Gallium-67, Gallium-68, Ruthenium 95, Ruthenium-97, Ruthenium-103, Ruthenium-105, Mercury-107, Mercury-203, Rhenium-99m, Rhenium-105, Rhenium-101, Tellurium-121m, Tellurium-122m, Tellurium-125m, Thulium-165, Thulium-167, Thulium-168, Technetium-99m and Fluorine-18.

Claim 14

Dependent Claim 12 words

The method of claim 1, wherein said radioisotope is Iodine-131 or Iodine-123.

Claim 15

Dependent Claim 34 words

The method of claim 1, wherein the amount of radiolabel introduced by direct chemical substitution into said marker-specific antibody fragment is sufficient to reduce its specific market substance immunoreactivity by from 5 to 33%.

Claim 16

Dependent Claim 38 words

The method of claim 1, wherein said marker-specific antibody fragment is substantially monospecific for the marker substance, having a crossreactivity with other antigens of less than 15% and an immunoreactivity for the marker substance of more than 50%.

Claim 17

Dependent Claim 28 words

The method of claim 1, wherein the specific antibody cleaved to produce said marker-specific fragment is a monoclonal antibody having a marker substance specificity of at least 90%.

Claim 18

Dependent Claim 42 words

The method of claim 1, wherein said fragment is labeled with a radioisotope of iodine, an average of at least 2.5 atoms of iodine per fragment molecule being introduced and no more than will reduce the antibody specificity by more than 33%.

Claim 19

Dependent Claim 22 words

The method of claim 1, wherein said tumor is located in the brain and said fragment is injected intravenously, intraarterially or intrathecally.

Claim 20

Dependent Claim 11 words

The method of claim 18, wherein the fragment is injected intravenously.

Claim 21

Dependent Claim 92 words

The method of claim 1, wherein prior to photoscanning, a second substance is injected into the subject, said second substance being radiolabeled with a radioisotope emitting at a different energy from the antibody fragment label and capable of independent detection by said photoscanning device, the level of activity of said second substance being used to determine the background activity due to non-targeted specific antibody fragment, said background activity being subtracted from the total activity of the specific antibody fragment, whereby the activity of substantially only the targeted, tumor-associated antibody fragment is determined.

Claim 22

Dependent Claim 24 words

The method of claim 20, wherein said second substance is technetium-99m-labeled normal human immunoglobulin, technetium-99m-labeled human serum, technetium-99m-sulfur colloid, technetium-99m-pertechnetate, or a combination thereof.

Claim 23

Dependent Claim 34 words

The method of claim 20, wherein said second substance is the corresponding fragment obtained by analogous cleavage of normal immunoglobulin from the same or different species as that used to prepare said specific antibody.

Claim 24

Dependent Claim 28 words

The method of claim 22, wherein said corresponding fragment is labeled with a different isotope of the same element used to label the fragment of said specific antibody.

Claim 25

Dependent Claim 52 words

The method of claim 23, wherein the specific antibody fragment is labeled with one of a pair of radioisotopes, and the corresponding normal immunoglobulin fragment is labeled with the other of the pair, said pair being Iodine-131 and Iodine-123; Indium-111 and Indium-113m; Gallium-67 and Gallium-68; Ruthenium-97 and Ruthenium-103; or Mercury-197 and Mercury-203.

Claim 26

Independent Claim 120 words

A method for detecting and localizing a tumor which either produces or is associated with a cytoplasmic, intracellular or cell-surface marker substance, which comprises injecting a human subject parenterally with two marker-specific fragments, one fragment being a marker-specific fragment obtained by cleavage of a first antibody specific to a first tumor associated marker and the second fragment being a marker-specific fragment obtained by cleavage of a second antibody specific to a second, different tumor-associated marker, each of said fragments being radiolabeled with a pharmacologically inert radioisotope capable of detection using a photoscanning device, and subsequently scanning with said device to detect and locate the site or sites of uptake of one or both of said labeled fragments by said tumor.

Claim 27

Independent Claim 120 words

A method for detecting and localizing a tumor which either produces or is associated with a cytoplasmic, intracellular or cell-surface marker substance, which comprises injecting a human subject parenterally with two marker-specific fragments, one fragment being a marker-specific fragment obtained by cleavage of a first antibody specific to a first tumor associated marker and the second fragment being a marker-specific fragment obtained by cleavage of a second antibody specific to a second, different tumor-associated marker, each of said fragments being radiolabeled with a pharmacologically inert radioisotope capable of detection using a photoscanning device, and subsequently scanning with said device to detect and locate the site or sites of uptake of one or both of said labeled fragments by said tumor.

Claim 28

Dependent Claim 26 words

The method of claim 25, wherein said two marker-specific fragments are obtained by cleavage of two of anti-CEA antibody, anti-AFP antibody, anti-HCG antibody and anti-CSAp antibody.

Claim 29

Dependent Claim 14 words

The method of claim 26, wherein said two fragments are labeled with different radioisotopes.

Claim 30

Dependent Claim 91 words

The method of claim 25, wherein prior to photoscanning, a reference substance is injected into the subject, said reference substance being radiolabeled with a radioisotope emitting at a different energy from either antibody fragment label and capable of independent detection by said photoscanning device, the level of activity of said reference substance being used to determine the background activity due to non-targeted specific antibody fragment, said background activity being subtracted from the total activity of each specific antibody fragment, whereby the activity of substantially only targeted, tumor-associated antibody fragments is determined.

Claim 31

Dependent Claim 24 words

The method of claim 28, wherein said reference substance is technetium-99m-labeled normal human immunoglobulin, technetium-99m-labeled human serum, technetium-99m-sulfur colloid, technetium-99m-pertechnetate, or a combination thereof.

Claim 32

Dependent Claim 112 words

The method of claim 28, wherein each of said two fragments is labeled with a different radioisotope; a fragment obtained by analogous cleavage of normal immunoglobulin from the same or different species used to prepare each said specific antibody and corresponding to each of said two fragments is used as a reference substance; each said reference fragment is labeled with a different isotope of the same element used to label the corresponding fragment of said specific antibody; and the activity of the reference fragment corresponding to each of said two fragments is used to determine the background activity and the targeted fragment activity for the specific antibody fragment to which it corresponds.

Claim 33

Independent Claim 122 words

A method for detecting and localizing a tumor which either produces or is associated with a cytoplasmic, intracellular or cell-surface marker substance, which comprises injecting a human subject parenterally with an F(ab') 2 hybrid formed from two chemically joined Fab' fragments, said fragments being a first marker-specific Fab' fragment obtained by cleavage of an antibody specific to a first tumor-associated marker and a second, different marker-specific Fab' fragment obtained by cleavage of a second antibody specific to the same or different tumor-associated marker, said hybrid being radiolabeled with a pharmacologically inert radioisotope capable of detection using a photoscanning device, and subsequently scanning with said device to detect and locate the site or sites of uptake of said labeled hybrid by said tumor.

Claim 34

Independent Claim 122 words

A method for detecting and localizing a tumor which either produces or is associated with a cytoplasmic, intracellular or cell-surface marker substance, which comprises injecting a human subject parenterally with an F(ab') 2 hybrid formed from two chemically joined Fab' fragments, said fragments being a first marker-specific Fab' fragment obtained by cleavage of an antibody specific to a first tumor-associated marker and a second, different marker-specific Fab' fragment obtained by cleavage of a second antibody specific to the same or different tumor-associated marker, said hybrid being radiolabeled with a pharmacologically inert radioisotope capable of detection using a photoscanning device, and subsequently scanning with said device to detect and locate the site or sites of uptake of said labeled hybrid by said tumor.

Claim 35

Dependent Claim 27 words

The method of claim 31, wherein said hybrid is obtained by cleavage and recombination of a mixture of F(ab') 2 fragments specific to each said specific antibody.

Claim 36

Dependent Claim 27 words

The method of claim 31, wherein said hybrid contains a marker-specific Fab' arm obtained by cleavage of anti-CEA antibody, anti-AFP antibody, anti-HCG antibody, or and anti-CSAp antibody.

Claim 37

Dependent Claim 30 words

The method of claim 31, wherein said hybrid contains marker-specific Fab' arms obtained by cleavage of two different antibodies selected from anti-CEA antibody, anti-AFP antibody, anti-HCG antibody and anti-CSAp antibody.

Claim 38

Dependent Claim 86 words

The method of claim 31, wherein prior to photoscanning, a second substance is injected into the subject, said second substance being radiolabeled with a radioisotope emitting at a different energy from the hybrid label and capable of independent detection by said photoscanning device, the level of activity of said second substance being used to determine the background activity due to non-targeted hybrid, said background activity being subtracted from the total activity of the hybrid, whereby the activity of substantially only the targeted, tumor-associated hybrid is determined.

Claim 39

Dependent Claim 24 words

The method of claim 35, wherein said second substance is technetium-99m-labeled normal human immunoglobulin, technetium-99m-labeled human serum, technetium-99m-sulfur colloid, technetium-99m-pertechnetate, or a combination thereof.

Claim 40

Dependent Claim 53 words

The method of claim 35, wherein said second substance is an F(ab') 2 fragment obtained by analogous cleavage of normal immunoglobulin from the same or different species as that used to prepare either said antibody specific to a first tumor-associated marker or said second antibody specific to the same or different tumor-associated marker.

Claim 41

Dependent Claim 26 words

The method of claim 37, wherein said second substance is labeled with a different isotope of the same element used to label the F(ab') 2 hybrid.

Claim 42

Dependent Claim 29 words

The method of claim 38, wherein the F(ab') 2 hybrid is labeled with Iodine-131 or Iodine-123 and the second substance is labeled with the other of Iodine-131 or Iodine-123.

Claim 43

Independent Claim 58 words

A method of tumor radiotherapy, which comprises parenterally injecting into a human subject having a tumor which produces or is associated with a cytoplasmic, intracellular or cell-surface marker substance an effective tumor reducing amount of a marker-specific fragment obtained by cleavage of an antibody specific to said marker substance and radiolabeled with a pharmaceutically inert, radiotherapeutically effective radioisotope.

Claim 44

Independent Claim 58 words

A method of tumor radiotherapy, which comprises parenterally injecting into a human subject having a tumor which produces or is associated with a cytoplasmic, intracellular or cell-surface marker substance an effective tumor reducing amount of a marker-specific fragment obtained by cleavage of an antibody specific to said marker substance and radiolabeled with a pharmaceutically inert, radiotherapeutically effective radioisotope.

Claim 45

Independent Claim 102 words

A method of tumor radiotherapy, which comprises parenterally injecting into a human subject having a tumor which either produces or is associated with at least one cytoplasmic, intracellular or cell-surface marker substance an effective tumor reducing amount of an F(ab') 2 hybrid formed from two chemically joined Fab' fragments, said fragments being a first marker-specific Fab' fragment obtained by cleavage of an antibody specific to a first tumor-associated marker and a second, different marker-specific Fab' fragment obtained by cleavage of a second antibody specific to the same or different tumor-associated marker, said hybrid being radiolabeled with a pharmacologically inert, radiotherapeutically effective radioisotope.

Claim 46

Independent Claim 102 words

A method of tumor radiotherapy, which comprises parenterally injecting into a human subject having a tumor which either produces or is associated with at least one cytoplasmic, intracellular or cell-surface marker substance an effective tumor reducing amount of an F(ab') 2 hybrid formed from two chemically joined Fab' fragments, said fragments being a first marker-specific Fab' fragment obtained by cleavage of an antibody specific to a first tumor-associated marker and a second, different marker-specific Fab' fragment obtained by cleavage of a second antibody specific to the same or different tumor-associated marker, said hybrid being radiolabeled with a pharmacologically inert, radiotherapeutically effective radioisotope.

Claim 47

Dependent Claim 93 words

The method of claim 41, wherein said F(ab') 2 hybrid is prepared by the steps of pepsin-catalyzed cleavage of anti-CEA antibody to produce an F(ab') 2 fragment and reductive cleavage thereof to form CEA-specific Fab' fragments, pepsin-catalyzed cleavage of anti-CSAp antibody to produce an F(ab') 2 fragment and reductive cleavage thereof to form CSAp-specific Fab' fragments, and oxidative coupling of a mixture of said CEA-specific Fab' fragments and sad CSAp-specific Fab' fragments to form F(ab') 2 fragments, at least a portion of which are F(ab') 2 hybrids specific to both CEA and CSAp.

Claim 48

Dependent Claim 28 words

The method of claim 42, wherein said F(ab') 2 hybrid is purified by sequential affinity chromatography through a column containing bound CEA and a column containing bound CSAp.

Claim 49

Dependent Claim 55 words

An injectable composition, which comprises: (a) a marker-specific fragment obtained by cleavage of an antibody specific to a cytoplasmic, intracellular or cell-surface marker substance which is provided or associated with a tumor, said fragment being radiolabeled with a pharmacologically inert radioisotope capable of detection using a photoscanning device; and (b) a pharmaceutically acceptable injection vehicle.

Claim 50

Dependent Claim 55 words

An injectable composition, which comprises: (a) a marker-specific fragment obtained by cleavage of an antibody specific to a cytoplasmic, intracellular or cell-surface marker substance which is provided or associated with a tumor, said fragment being radiolabeled with a pharmacologically inert radioisotope capable of detection using a photoscanning device; and (b) a pharmaceutically acceptable injection vehicle.

Claim 51

Dependent Claim 41 words

The composition of claim 44, which further comprises a second marker-specific fragment obtained by cleavage of a second antibody specific to a second tumor-associated marker, said fragment being radiolabeled with a pharmacologically inert radioisotope capable of detection using a photoscanning device.

Claim 52

Dependent Claim 70 words

The injectable composition of claim 44, which further comprises a reference fragment which is the corresponding fragment obtained by analogous cleavage of normal immunoglobulin from the same or different species as that used to prepare said specific antibody, said reference fragment being radiolabeled with a different isotope of the same element used to label said marker-specific fragment and emitting at an energy capable of independent detection using said photoscanning device.

Claim 53

Dependent Claim 83 words

An injectable composition which comprises: (a) an F(ab') 2 hybrid formed from two chemically joined Fab' fragments, said fragments being a first marker-specific Fab' fragment obtained by cleavage of an antibody specific to a first tumor-associated marker and a second different marker-specific Fab' fragment obtained by cleavage of a second antibody specific to the same or different tumor associated marker, said hybrid being radiolabeled with a pharmacologically inert radioisotope capable of detection using a photoscanning device; and (b) a pharmaceutically acceptable injection vehicle.

Claim 54

Dependent Claim 83 words

An injectable composition which comprises: (a) an F(ab') 2 hybrid formed from two chemically joined Fab' fragments, said fragments being a first marker-specific Fab' fragment obtained by cleavage of an antibody specific to a first tumor-associated marker and a second different marker-specific Fab' fragment obtained by cleavage of a second antibody specific to the same or different tumor associated marker, said hybrid being radiolabeled with a pharmacologically inert radioisotope capable of detection using a photoscanning device; and (b) a pharmaceutically acceptable injection vehicle.

Claim 55

Dependent Claim 63 words

The injectable composition of claim 47, which further comprises a reference F(ab') 2 fragment from normal immunoglobulin from the same or different species as that used to prepare said hybrid, said reference fragment being radiolabeled with a different isotope of the same element used to label said F(ab') 2 hybrid and emitting at an energy capable of independent detection using said photoscanning device.

Claim 56

Independent Claim 114 words

A method of tumor radiotherapy, which comprises parenterally injecting into a human subject having a tumor which produces or is associated with a cytoplasmic, intracellular or cell-surface marker substance an effective tumor reducing amount of a marker-specific fragment obtained by cleavage of an antibody specific to said marker substance and radiolabeled with a pharmacologically inert radioisotope capable of detection with a photoscanning device, said labeled fragment further containing in chemical combination an addend containing at least five atoms of boron with at least a natural abundance of Boron-10 isotope; wherein the location of said tumor is determined using said photoscanning device, and a beam of thermal neutrons is then directed at said tumor location.

Claim 57

Independent Claim 114 words

A method of tumor radiotherapy, which comprises parenterally injecting into a human subject having a tumor which produces or is associated with a cytoplasmic, intracellular or cell-surface marker substance an effective tumor reducing amount of a marker-specific fragment obtained by cleavage of an antibody specific to said marker substance and radiolabeled with a pharmacologically inert radioisotope capable of detection with a photoscanning device, said labeled fragment further containing in chemical combination an addend containing at least five atoms of boron with at least a natural abundance of Boron-10 isotope; wherein the location of said tumor is determined using said photoscanning device, and a beam of thermal neutrons is then directed at said tumor location.

Claim 58

Dependent Claim 33 words

The injectable composition of claim 44, wherein said marker-specific fragment further contains in chemical combination an addend containing at least five atoms of boron with at least the natural abundance of Boron-10 isotope.

Annuity Stage	Window Opens	Payment Deadline	Surcharge Period Until	Estimated Fee	Status
3.5 Years (Year 4)	1985-05-24	1985-11-23	1986-05-25	$2000.00	Window Closed / Lapsed
7.5 Years (Year 8)	1989-05-24	1989-11-23	1990-05-25	$3760.00	Window Closed / Lapsed
11.5 Years (Year 12)	1993-05-24	1993-11-23	1994-05-25	$7700.00	Window Closed / Lapsed

Tumor localization and therapy with labeled antibody fragments specific to tumor-associated markers

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