Neorx Corporation

Daniel S Wilbur

Alan R Fritzberg

Ananthachari Srinivasan

Sudhakar Kasina

Stephen W Hadley

David S Jones

Mark D Hylarides

Dennis W Wester

Protein conjugated chelated metal radionuclides are provided for use in vivo. Intermediates are provided for preparing the polypeptide compositions efficiently.

Metal radionuclide labeled proteins for diagnosis and therapy

Haloaryl compounds are lithiated and thereafter metalated with one of the following organometallic groups: Sn(n-Bu).sub.3 or SnMe.sub.3. The resulting aryltin compound can be transmetalated in site-specific reaction with one of the following organometallic groups: HgX, Hg(OAc).sub.2, BX.sub.3, or BZ.sub.2, wherein X is Cl, Br, or I, and Z is alkyl or alkoxy. The metalated compounds are subsequently radiohalogenated via a demetalation reaction. A functional group suitable for conjugation to protein can be added subsequent or preferably prior to the radiohalogenation. Also compounds of the formula: R.sub.1 -Ar-R.sub.2, wherein R.sub.1 is either a radiohalogen or any one of the organometallic groups stated above, Ar is aromatic or heteroaromatic ring, and R.sub.2 is a short-chain substituent that does not activate the aromatic ring and that bears a functional group, or a precursor thereof, suitable for conjugation to protein under conditions that preserve the biological activity of the protein. The radiohalogenated small molecules are conjugated to proteins such as monoclonal antibodies for use in diagnosis and therapy.

Radiohalogenated small molecules for protein labeling

Vinyl radiohalogenated small molecules as shown in formulas I and II: ##STR1## wherein *X is radiohalogen, C.dbd.C is a double bonded set of sp.sup.2 hybridized carbon atoms, and substituents R.sub.1, and R.sub.2 are as defined in the specification. Y is a substituent containing any of the groups described for R.sub.1 and R.sub.2, except that Y cannot be hydrogen, and bearing a functional group suitable for binding to protein under conditions that preserve the biological activity of the protein. The compounds of formulas I and II can be coupled to proteins such as monoclonal antibodies to provide reagents for diagnostic and therapeutic applications. Also metalated precursors of compounds I and II, as well as radiopharmaceutical reagent kits containing any of the subject small molecules.

Vinyl substituted radiohalogen conjugates for protein labeling

Method of producing metal radionuclide labeled proteins for diagnosis

A process for recovering At-211 from a target and simultaneously producing an At-211-labeled radiopharmaceutical agent involves introducing volatilized At-211, produced by dry distillation of an irradiated Bi-209 target, into a solution containing a compound that binds the At-211 to form an At-211-labeled radiopharmaceutical agent. The radiopharmaceutical agent may have therapeutic use as is, or may be attached to a targeting protein such as an antibody prior to administation to a patient.

Process for isolation and radiolabeling of pharmaceuticals with isotopes

There is disclosed a process for minimally derivatizing a targeting protein with a radionuclide such that the predominant species of derivatized targeting protein contains one radionuclide group as a single radiolabeled ligand, and whereby the binding characteristics of the targeting protein are minimally affected. There is further disclosed ligands that can chelate a metal radionuclide or bind a halogen radionuclide while providing a means for separating radiolabeled ligand from unradiolabeled ligand.

Minimal derivatization of proteins

Modified cellular substrates are used as linking groups that are recognized as cellular substrates by intracellular metabolic enzymes. The linking groups function to attach a ligand, such as a drug or radionuclide, to a targeting protein, such as an antibody or antibody fragment. The false substrate structure of the linking groups causes increased cellular retention of the linking group/ligand conjugate because the enzymes used for catabolism of the substrate are inhibited by the false substrate structure of the linking group.

Modified cellular substrates used as linkers for increased cell

Haloaryl compounds are lithiated and thereafter metalated with one of the following organometallic groups: Sn(n-Bu).sub.3 or SnMe.sub.3. The resulting aryltin compound can be transmetalated in site-specific reaction with one of the following organometallic groups: HgX, Hg(OAc).sub.2, BX.sub.3, or BZ.sub.2, wherein X is Cl, Br, or I, and Z is alkyl or alkoxy. The metalated compounds are subsequently radiohalogenated via a demetalation reaction. A functional group suitable for conjugation to protein can be added subsequent of preferably prior to the radiohalogenation. Also compounds of the formula: R.sub.1 -Ar-R.sub.2, wherein R.sub.1 is either a radiohalogen or any one of the organometallic groups stated above, Ar is aromatic or heteroaromatic ring, and R.sub.2 is a short-chain substituent that does not activate the aromatic ring and that bears a functional group, or a precursor thereof, suitable for conjugation to protein under conditions that preserve the biological activity of the protein. The radiohalogenated small molecules are conjugated to proteins such as monoclonal antibodies for use in diagnosis and therapy.

Haloaryl compounds bearing a defined functional group or precursor are metalated with either Sn(n-Bu).sub.3 or SnMe.sub.3. The resulting aryltin compound may be transmetalated in site-specific reaction with one of the following organometallic groups: HgX.sub.2, Hg(OAc).sub.2, BX.sub.3, or BZ.sub.3, wherein X is Cl, Br, or I, and Z is alkyl or alkoxy. The stannylated or otherwise metalated compounds are subsequently radiohalogenated via a demetalation reaction. The Stannylated or otherwise metalated compounds are subsequently radiohalogenated via a demetalation reaction. The functional group is suitable for conjugation to protein and can be present or be added subsequent, but most preferably prior, to the radiohalogenation. Also compounds of the formula: R.sub.1 --Ar--R.sub.2, wherein R.sub.1 is either a radiohalogen or any one of the organometallic groups stated above, Ar is aromatic or heteroaromatic ring, and R.sub.2 is a short-chain substituent that does not highly activate the aromatic ring an that bears a functional group suitable for conjugation to protein under conditions that preserve the biological activity of the protein. The radiohalogenated small nolecules are conjugated to proteins such as monoclonal antibodies for use in diagnosis and therapy.

Radiohalogenated for proteins

Vinyl radiohalogenated small molecules as shown in formulas I and II: ##STR1## wherein *X is a radiohalogen, C.dbd.C is a double bonded set of sp.sup.2 hybridized carbon atoms, and substituents R.sub.1, R.sub.2, and Y are as defined below. R.sub.1 and R.sub.2 are substituents independently selected from among hydrogen; alkyl or substituted alkyl, provided that any sp.sup.2 or sp carbon atom substituted on the alkyl is separated from C.dbd.C by at least one fully substituted sp.sup.3 carbon atom; aryl or substituted aryl, provided that the aryl is bonded directly to C.dbd.C; heteroalkyl, provided, first, that no heteroatom of the heteroalkyl bonds directly to C.dbd.C and, second, that any sp.sup.2 or sp hybridized carbon bonded to a heteroatom of the heteroalkyl is not bonded directly to or otherwise conjugated with C.dbd.C and, third, that where a single sp.sup.3 carbon intervenes between C.dbd.C and an sp.sup.2 or sp carbon bonded to a heteroatom that intervening sp.sup.3 carbon must be fully substituted; heteroaryl, provided that a heteroatom of the heteroaryl is not bonded directly with C 50 C; mixed alkylaryl, provided first, that no heteroatom is bonded to C.dbd.C and, second that either an aryl moiety of the mixed alkylaryl is directly bonded to C.dbd.C or that any aryl moiety is separated from C.dbd.C by at least one sp.sup.3 carbon atom and, where only one sp.sup.3 hybridized carbon atom intervenes between C.dbd.C and an aryl moiety, that intervening sp.sup.3 carbon must be fully substituted. Y is substituent containing any of the groups described above for R.sub.1 and R.sub.2, except that Y cannot be hydrogen, and bearing a functional group suitable for binding to protein under conditions that preserve the biological activity of the protein. The compounds of formulas I and II can be coupled to proteins such as monoclonal antibodies ot provide reagents for diagnostic and therapeutic applications. Also metalated precursors of compounds I and II, as well as radiophormaceutical reagent kits containing any of the subject small molecules.

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Edmonds, WA, United States of America

Daniel S Wilbur