AU621585B2 – Improvements in or relating to modified haptens useful as imaging and therapeutic agents
– Google Patents
AU621585B2 – Improvements in or relating to modified haptens useful as imaging and therapeutic agents
– Google Patents
Improvements in or relating to modified haptens useful as imaging and therapeutic agents
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Publication number
AU621585B2
AU621585B2
AU29563/89A
AU2956389A
AU621585B2
AU 621585 B2
AU621585 B2
AU 621585B2
AU 29563/89 A
AU29563/89 A
AU 29563/89A
AU 2956389 A
AU2956389 A
AU 2956389A
AU 621585 B2
AU621585 B2
AU 621585B2
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AU
Australia
Prior art keywords
antibody
hapten
edta
dtpa
branched
Prior art date
1988-02-03
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Application number
AU29563/89A
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AU2956389A
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Inventor
Leslie D Anderson
Thomas Robert Battersby
James M. Frincke
Edward Kusnierz
Damon L. Meyer
Brian William Sullivan
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Hybritech Inc
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Hybritech Inc
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1988-02-03
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1989-02-02
Publication date
1992-03-19
1989-02-02
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Hybritech Inc
1989-08-03
Publication of AU2956389A
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patent/AU2956389A/en
1992-03-19
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1992-03-19
Publication of AU621585B2
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Critical
patent/AU621585B2/en
2009-02-02
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Classifications
C—CHEMISTRY; METALLURGY
C07—ORGANIC CHEMISTRY
C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
C07C335/00—Thioureas, i.e. compounds containing any of the groups, the nitrogen atoms not being part of nitro or nitroso groups
C07C335/04—Derivatives of thiourea
C07C335/16—Derivatives of thiourea having nitrogen atoms of thiourea groups bound to carbon atoms of six-membered aromatic rings of a carbon skeleton
C07C335/22—Derivatives of thiourea having nitrogen atoms of thiourea groups bound to carbon atoms of six-membered aromatic rings of a carbon skeleton being further substituted by carboxyl groups
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
A61K51/0474—Organic compounds complexes or complex-forming compounds, i.e. wherein a radioactive metal (e.g. 111In3+) is complexed or chelated by, e.g. a N2S2, N3S, NS3, N4 chelating group
A61K51/0478—Organic compounds complexes or complex-forming compounds, i.e. wherein a radioactive metal (e.g. 111In3+) is complexed or chelated by, e.g. a N2S2, N3S, NS3, N4 chelating group complexes from non-cyclic ligands, e.g. EDTA, MAG3
A61K51/048—DTPA (diethylenetriamine tetraacetic acid)
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
A61K51/1093—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 conjugates with carriers being antibodies
A—HUMAN NECESSITIES
A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
A61P35/00—Antineoplastic agents
C—CHEMISTRY; METALLURGY
C07—ORGANIC CHEMISTRY
C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
C07C237/00—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by amino groups
C07C237/02—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by amino groups having the carbon atoms of the carboxamide groups bound to acyclic carbon atoms of the carbon skeleton
C07C237/04—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by amino groups having the carbon atoms of the carboxamide groups bound to acyclic carbon atoms of the carbon skeleton the carbon skeleton being acyclic and saturated
C07C237/06—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by amino groups having the carbon atoms of the carboxamide groups bound to acyclic carbon atoms of the carbon skeleton the carbon skeleton being acyclic and saturated having the nitrogen atoms of the carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms
C—CHEMISTRY; METALLURGY
C07—ORGANIC CHEMISTRY
C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
C07C323/00—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups
C07C323/50—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and carboxyl groups bound to the same carbon skeleton
C07C323/51—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and carboxyl groups bound to the same carbon skeleton having the sulfur atoms of the thio groups bound to acyclic carbon atoms of the carbon skeleton
C07C323/60—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and carboxyl groups bound to the same carbon skeleton having the sulfur atoms of the thio groups bound to acyclic carbon atoms of the carbon skeleton with the carbon atom of at least one of the carboxyl groups bound to nitrogen atoms
C—CHEMISTRY; METALLURGY
C07—ORGANIC CHEMISTRY
C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
C07C335/00—Thioureas, i.e. compounds containing any of the groups, the nitrogen atoms not being part of nitro or nitroso groups
C07C335/04—Derivatives of thiourea
C07C335/16—Derivatives of thiourea having nitrogen atoms of thiourea groups bound to carbon atoms of six-membered aromatic rings of a carbon skeleton
C07C335/20—Derivatives of thiourea having nitrogen atoms of thiourea groups bound to carbon atoms of six-membered aromatic rings of a carbon skeleton being further substituted by nitrogen atoms, not being part of nitro or nitroso groups
A—HUMAN NECESSITIES
A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
A61K2121/00—Preparations for use in therapy
A—HUMAN NECESSITIES
A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
A61K2123/00—Preparations for testing in vivo
Abstract
The present invention discloses a method for modifying the pharmacokinetics of a hapten which is useful as an in vivo radioimaging or radiotherapeutic agent. Compositions comprising a derivatized hapten of the invention are provided for in vivo imaging and therapy. A compound of Formula (I): p-R1-C6H4-CH2-EDTA-M (I) wherein R1is -NH- @-NH-R2, – @-NH-R2, -NH- @-CH2-R2, -NH- @-CH2-NH-R2, -NH-R2, -NH- @-NH-R2 or -NH- @-CH2-S-R2; R2 is hydrogen, an amino group, -OC @Y), -p-phenyl-CH2-Y, an unsubstituted C1 to C30 branched or straight chain alkyl group; a substituted C1 to C30 branched or straight chain alkyl, cycloalkyl, aryl or arylalkyl group, in which the substituents are one or more of any of: hydroxy, carboxy, =O, =S, – @=O, – @=S -S-, -SR4, fluoro, chloro, bromo, iodo, amino, nitro, -SO3H, -NHR3, -NHR4, -N(R3)2, -CONHR3, -COOR3, -SO4, -PO4, phenyl, benzyl, imidazolo, or a group of the formulae: -NH- @@NH-R3, -NH- @-NH-R3 or NH- @-NH-R3; wherein R3 is hydrogen, a C1 to C30 straight or branched chain alkyl group, -p-phenyl-CH2-Y, – @-CH3, or a non-reactive functional group; Y is EDTA, DTPA, DOTA, HETA, TRITA or TETA; R4 is H or -CH2- @-NHR3; and, M is a metal ion, or a pharmaceutically-acceptable salt thereof.
Description
L ;-rut G< I Colin W. Ewing (Signature of Declarant) General Counsel and Secretary TO: THE COMMISSIONER OF PATENTS
AUSTRALIA
S&F REF: 83244 eah:9472D
U::
L4 )I ii I~ 1 'LL- 1 -u IIJU C- ~L P~ 9 i 621585 S F Ref: 83244 FORM COMMONWEALTH OF AUSTRALIA PATENTS ACT 1952 .4 COMPLETE SPECIFICATION
(ORIGINAL)
FOG OFFICE USE: Class Int Class Complete Specification Lodged: Accepted: Published: Priority: Related Art: Name and Address of Applicant: Hybritech Incorporated 11095 Torreyana Road San Diego California 92121 UNITED STATES OF AMERICA Address for Service: L I Spruson Ferguson, Patent Attorneys Level 33 St Martins Tower, 31 Market Street Sydney, New South Wales, 2000, Australia Complete Specification for the invention entitled: Improvements in or Relating to Modified Haptens Useful as Imaging and Therapeutic Agents The following statement is a full description of this invention, best method of performing it known to me!us including the 5845/3 L i 1 H-7586 -1- IMPROVEMENTS IN OR RELATING TO MODIFIED HAPTENS USEFUL AS IMAGING AND THERAPEUTIC AGENTS .9 0 S S 0* .9
S
9
S
9. 9 09 05 9*9 0 9. 9 6 59 S 9 9. S Monoclonal antibodies are becoming increasingly important for in vivo use. Of substantial interest are their potential applications for the imaging and treatment of disease, particularly cancer.
Monoclonal antibodies which are labeled, for example, with a chelate complex of a metal ion and a chelating 10 agent are particularly useful for the imaging and treatment of certain disease states. Generally, such monoclonal antibodies are labeled by chemical conjugation with the hapten or, in the case of anti-hapten antibodies, by their ability to specifically bind with 15 the metal chelate to form non-covalent hapten-antibody complexes.
Recent studies relating to the use of monoclonal antibodies for tumor imaging and therapy have shown that bifunctional antibodies having specificity for a hapten and, for example, a tumor target, provide a mechanism for delivery of the hapten to the tumor target. Delivery of a hapten to a disease site with a bifunctional antibody system involves time dependent binding and release of the hapten from the antibody.
Consequently, the usefulness of a particular hapten as an imaging or therapeutic agent is dependent upon the interaction of the hapten with the antibody utilized -s well as with serum components, proteins, lipids, and other tissue compartments. For example, the inherent pharmacokinetic properties of a particular H-7586 -2hapten and the affinity of an antibody for the hapten may result in the hapten binding so tightly to the anti-hapten antibody so as to be indistinguishable from covalently bound antibody conjugates, or in the hapten binding so loosely to the anti-hapten antibody so as to be incapable of localization with the antibody at the disease site. Additionally, the inherent pharmacokinetic properties of a particular hapten may result in 16 .undesirable serum and tissue interactions such that the 10 accumulation of the hapten in the liver, kidneys or intestines reaches intolerable levels. In such cases, the biodistribution and localization of the hapten substantially limits the potential clinical value of the bifunctional antibody delivery system.
S 15 Accordingly, there exists a need to modify pharmacokinetic properties of various haptens for in vivo imaging and therapy. Further, there exists a need i for novel hapten derivatives that impart desired proper- 9 ties to antibody delivery systems for particular appli- 20 cations. The present invention meets these needs by providing-novel hapten derivatives with modified pharmacokinetics, as well as methods for modifying the pharmacokinetics of a given hapten.
Surprisingly, haptens which are useful as in vivo imaging and therapeutic agents can be derivatized to modify the inherent pharmacokinetic properties of the hapten. Accordingly, besides providing4novel hapten derivatives, the present invention provides a method for modifying the pharmacokinetics of a hapten as a radioimaging or radiotherapeutic agent which comprises ;l I. t }J 1 H-7586 -3selecting a suitable hapten and an antibody capable of complexing with the hapten, and derivatizing the hapten to modify the dissociation rate of the hapten-antibody complex and the interaction of the hapten with serum components and tissue compartments, thereby increasing or decreasing serum half-life and radiation doses to tissues. A predetermined effective dosage of the derivatized hapten and the antibody are thereafter administered to a patient for purposes of imaging or therapy.
Haptens which can be derivatized, in accor- 1 dance with the present invention, are generally selected *from complexes of a chelating agent and a metal ion.
Preferably, the hapten selected for use in the invention is a metal ion complex of ethylenediaminetetracetic acid diothylenetriaminepentaacetic acid ("DTPA"), F 1,4,7,10-tetraazacyclododecane-N,N',N'',N ''-tetraacetic acid 1,5,9,13-tetraazacyclohexadecane- II'-tetraacetic acid ("HETA"I), 1,4,7,10,tetraazacyclotridecane-N,N',N'",N'' -tetraacetic acid ("TRITA") or 1,4,8,11-tetraazacyclotetradecane- N,N' -tetraacetic acid ("TETA") with the derivatized hapten being represented generally by Formula p-R 1
-C
6 H4-CH 2 -EDTA-M (I) S 0 wherein R 1 is -NH-C-NH-R 2 -C-Nd-R 2
-NH-C-CH
2
-R
2
-NH-C-CH
2
-NH-R
2 II II o 0 -4- NHCNH-R2, or -NH-C-CH2-S-R2; 0 0 R2 is hydrogen, an amino group, -0C44-) -p-phenyl-CH 2 an unsubstituted C 1 I to C 30 branched or straight chain alkyl group; a substituted C 1 to C 30 branched or straight chain alkyl, cycloalkyl, aryl or arylalkyl group, in which the substituents are one or more of any of: hydroxy, carboxy, -C=0
R
3 -S fluoro, chioro, bromo, iodo, amino, nitro, -SO 3 H, -NHRS, -NHR, -N(R CONHR 3
-COOR,
-so 4 P phenyl, benzyl, imidazolo, or a group of the formulae:
-NH-C-NHR
3 -NH-C-NH-R, -NH-C- NH-R; 11 3 f 11 3t 11 3 NH 0 S wherein R is hydrogen, a C 1 to C 0 straight or branched canalkyl group, -p-phenyl -Y,
-C-CH;
3.II Y is EDTA, DTPA, DOTA, HETA, TRITA or TETA /jjA
R
4 is H or -CH 2
-C-NHR
3 and, M is a metal ion 0 In a preferred embodiment, diagnostic or therapeutic formulations wherein the active ingredients are compounds of Formula (Ia): p-R 1
-C
6
H
4
CH
2 EDTA-M (Ia) S 0 0 II II Il wherein R 1 is -NH-C-NHR 2
-C-NH-R
2 or -NH-C-CH 2
-R
2 in which R 2 is hydrogen; NH 2 an unsubstltuted C 1
-C
30 S branched or straight chain alkyl group; or a C1-C30 branched or straight chain alkyl, cycloalkyl, aryl or arylalkyl group substituted by one or more substituents independently selected from the group consisting of: -OH; -COOH; -Cl; -Br; -NH 2
-NO
2 -S0 3 H; -NH-R 3 -CONHR3; -COOR 3 -S04; -P0 4 phenyl; and benzyl; R is H; or an alkyl group; and M is a metal ion, are useful as imaging or therapeutic agents.
Especially preferred compounds useful in the invention are S" those represented by Formula (Ib): p-R 1
-C
6 H4-CH 2 -EDTA-M (Ib) wherein R1 is -NH- -NH-R 2 -NH- -CH 2
-S-R
2
-NH-C-CH
2
-R
2 0 0 0
-NH-R
2
-NH-C-CH
2
-NH-R
2 or -NH- -NH-R 2 0 S .08 I 't R I ,j R2 is -p-phenyl-CH 2 -OC a substituted C 1 to C 30 branched or straight chain alkyl,.
cycloalkyl, aryl or arylalkyl group, in which the substituents are one or more of any of:
R
3
R
3 hydroxy,
-SR
4 -NHR 3 -NHR 4
N(R
3 2 imidazolo, or a group of the formulae: -NH- NH-R 3 NH-C-NH-R 3 or -NH-C--NH-R 3 NH 0 S wherein R 3 is p-phenyl-CH 2
-Y,
-C-CH;
0 Y is EDTA, DTPA, DOTA, HETA, TRITA or TETA; Ri3~ H or -CH 2 -C-NHR and, 0 M is a metal ion, Any one of the above formulations of the invention may further comprise, separate from or in combination with a hapten of the invention, an antibody capable of complexing with the hapten.
4308a/jj H-7586 -7- Further, the present invention provides methods for in vivo imaging and therapy which comprises administering to a patient, sequentially, simultaneously, or in combination, a predetermined effective dosage of a hapten of the invention, and a suitable anti-hapten antibody.
The invention has been summarized in order that the detailed description that follows may be better understood and the contribution to the art may be better appreciated.
As indicated above, the present invention, in one aspect, provides a method for modifying the pharmacokinetics o;f a hapten as an in vivo radioimaging or radiotherapeutic agent. The method comprises the selection of a hapten suitable for imaging or therapy and an antibody capable of complexing with the hapten, and the derivatization of the hapten to modify the dissociation rate of the hapten-antibody complex and the e* interaction of the hapten with serum and tissue .o 20 components, thereby increasing or decreasing serum half-life, as desired. The derivatized hapten and the antibody are administered to a patient in a predetermined effective doAge for imaging or therapy.
Preferably, the biodistribution and localization of the derivatized hapten, and complexes of the derivatized hapten and the antibody, are enhanced depending upon the particular diagnostic or therapeutic purpose.
"In the context of the present invention, the term "hapten" refers to a molecule capable of specific reactivity with an antibody, but incapable of H-7586 -8stimulating an immune response by antibody production, except in combination with or as part of a carrier protein molecule. As used herein, the term "pharmacokinetics" refers to the change in the biodistribution of a molecule over a period of time.
The terms "biodistribution" and "localization," as used herein, refer to the distribution of a molecule within individual organs of an animal at given time-points, and the amount or concentration of a molecule present in an S 10 organ, respectively. Further, the term "localization" implies the presence, in excess of the concentration due to blood levels, of a molecule at a particular organ for a period of time as a result of the interaction between the molecule and tissue compartments.
For purposes of the present invention, the hapten selected for use in the invention will generally depend upon the chemical, biological and physiological properties of the hapten and the desired application of the invention. For example, the selection of a S: 20 particular hapten will depend upon whether the hapten will be used as an imaging or therapeutic agent, and generally involves consideration of chemical properties, such as water solubility, biological properties such as the rate of clearance from particular tissues, and physiological properties, such as the capability of binding to a receptor at a disease site.
Haptens comprising chelate complexes of a metal ion and a chelating agent are generally selected for use in the invention. Preferred chelating agents of the invention are ethylenediaminetetracetic acid i 1- H-7586 -9diethylenetriaminepentaacetic acid ("DTPA"), DOTA, HETA, TRITA, TETA, and analogs thereof. EDTA, DTPA, and methods for preparing analogs and the starting materials for the compounds of the present invention are Cdscribed in U.S. Patent No. 4,622,420. The methods for preparing DOTA, HETA, TRITA, TETA, and starting materials useful in the present invention are described in detail in U.S. Patent No. 4,678,667 (Meares, et al.) and in Moi, et al., J. Am. Chem. Soc., 110, 6266 (1988), 10 the teachings of both of which are incorporated herein S. by reference. The EDTA and DTPA analogs disclosed in U.S. Pat. No. 4,622,420 are capable of forming physiologically stable chelates with a variety of metal ions.
Likewise, the DOTA, HETA, TRITA, and TETA analogs
I
15 descriDed in U.S. Pat. No. 4,678,667 and Moi, et al.
are capable of forming physiologically stable chelates with various metal ions.
As used in Formulae and when referring to a metal ion means a metal atom in 20 ionic form without particular reference to possible valences. The particular valence of the ion, as one skilled in the are will recognize, depends on the particular metal in question. Thus, the term is not St meant to limit the metal ion to any particular state of valency. The preferred metal ions, as indicated below, however, are 90 y3+ and 1 llIn 3 Preferred for use in the present invention are haptens that form chelate complexes with metal ions generally selected from indium-lll ("11 1 technetium-99m 9 9 mTc"), copper-67 6 7 Cu") gallium-67 H-7586 preferred for diagnostic application, i.e. imaging, while is generally preferred for therapeutic application. Additionally, the selection of a suitable metal ion may involve consideration of the affinity the metal complex has for the anti-hapten antibody utilized in the invention as different metal complexes exhibit different affinities for certain anti-hapten antibodies.
SThe compoundsA;; the invention having designations DB-I to DB-VIII are particularly useful in overcoming these latter difficulties associated with an anti-hapten antibody showing different affinities for different metal complexes. These particular haptens of the invention possess a unique "dumbbell" shape (thus, the designation with two separate -10-1 moieties capable of chelating metal ions used in the invention. For example, the DTPA moiety ccldr~ complex, for example, 1 llIn, the prpferred S metal ion for diagnostic purposes. Once the diagnostic procedure is complete, the same hapten can be used to complex, at the DTPA moiety, a metal Ion, for example, 90 useful for therapeutic applications. Thus, the problem of different anti-hapten antibody affinities for different hapten-metal ion complexes could be alleviated or overcome. A single hapten could be used for both d agnostic and therapeutic applications, The preferred haptensf the invention comprise benzyl EDTA derivatives.
Such haptens contain structural moieties that can conveniently complex a metl cmplxes Thse prtiula haten ofthe nvetio poses a
-J
varet ofmealions, including ill n and 90Y or the desired diagnostic or therapeutic application. Benzyl EDTA, analogs thereof, and methods for their preparation are described in U.S. Pat. No. 4,622,420. In accordance with the invention, such haptens are derivatized to have a structure represented by Formula (ID: p-R 1
-C
6
H
4
CH
2 -EDTA-M (I) S 0 0 11 11 11 wherein RIis -NfI-C-NH-R 2 1 -NH-C-NH-R 2
-C-NH-R,
**-NH-c-CH 2 R 2
NH-G-CH
2
NH-R,
2 2f *0 0
~-NH-R
2 or -NH-C-CH 2 2 R2 is hydrogen, an amino group, ~-p-phenyl-CH 2 an unsubstituted C 1 to C 30 branched or straight chain alkyl group; a substituted C 1 I to C 30 branched or straight chain S alkyl, cycloalkyl, aryl or arylalkyl group, in which the substituents axe one or more of any of: hydroxy, carboxy, -C=S, I I
K
3 KR 3
-SR
4 fluoro, chloro, bromo, iodo, amino, nitro, -So 3 H, -NHR 3 1 NHR 4
-N(R
3 2
-CONHR
3 COOR 3 1 -SO 4 -P0 4 phenyl benzyl imidazolo, or a group of the formulae: -NH-C-NH-R3, NH-C-NH-R -NHI-C-NH-R; NH 0 S 8adj nyaroxy, carboxy,
-C=Q,
-SR 4 fluoro, chloro, bromo, lodo,
"I
11 R 3 R3 2 r A 9 9 9 9* 9 9.
*99* #999*9 9 13- Additionally, R 2 may Further comprise non-reactive functie-Kial groups which may include substituents that exhibit specific tissue interactions, ,l acids, polypeptides, nucleotides, polynucleotides, carbohydrates or lipids. As used for R2and R 3 the term "non-reactive functional group" refers to a functional group incapable of forming a covalent bond with tissue or serum components, and the anti-hapten antibody utilized, under physiological conditions.
Especially preferred compounds useful in the invention are those repre-sented by Formula (Ib): p-R I- C 6 6 4 CH 2 "EDTA-M (1b) wherein Rl s -H-CNHR 2 -NH-C-CH 2 S-R 2 NH-C-CH 2
-R
2
-NIH-R
2 -NH-G-CH -NH-R, or -NH-C-NH-R; 0 S RZ is -p-phenyl-CH 2 -OC a substitote C 1 to C 30 branched or straight chain alkyl, cycloalkyl, aryl or arylalkyl group, in which the substituents are one or more of any of:
R
3
R
3 I I hydroxy, -SR 4
NHR
3
-NHR
4 -NR )21or a group of the formulae: NH -C-NH-R N-5N- H N- 11 R 3 7 -NHc-N- 31 -NH.?1H 3 NH 0 S 4 a. t, a a a.
d 0 0* a. a S. S as a.
OJ 0 *4 a I .d S 9, *1 0* .J S.
H- 7586 -14wherein
R
3 is -p-pheny1-CH 2
-Y,
or
-C-CH
3 11 0 Y is EDTA, DTPA, DOTA, HETA, TRITA or TETA;
-R
4 is H or -CH 2
-C-NHR
3 and, M is a metal ion.
11 0
\AS,
In one embodiment, preferred compounds\.f the invention are p-thioureidobenzyl EDTA derivatives or 15 analogs thereof, which are derived from isothiocyanatobenzyl EDTA. Isothiocyanatobenzyl RDTA and isothiocyanatobenzyl DTPA, analogs thereof, and methods for their preparation are described in U.S. Patent No. 4,622,420.
Such preferred haptens of the invention, have a struc- 20 ture represented by the following formula:
S
11 p-R 2
-NH-C-NH-C
6
H
4
-CH
2
-EDTA-M
25 whierein R 2 is hydrogen; NH 2 an unsubstituted
C
1 branched or straight chain alkyl group; or a
CI-C
18 branched or straight chain alkyl, cycloalkyl, aryl or arylalkyl group, substituted by one or more substituents independently selected from the group consisting of: -OH; -NH 2 -COOH; 2 phenyl; benzyl; -S0 3 H; and -NO 2 and M is a metal- ion.
q "'0Vi (StC($ 5845/3
I;
;i j bl i 64
J
H-7586 In addition, '2 may further comprise non-reactive functional groups including substituents that may exhibit specific tissue interactions, amino acids, polypeptides, nucleotides, polynucleotides, carbohydrates, or lipids.
In Formulae and the designation is meant to relate to one skilled in the art that the moiety as defined in the formulae, is linked to the adjacent portion of the molecule through 10 one of the carboxy groups of the Y moiety. On the other hand, the designation alone, is meant to indicate that the Y moiety is linked to the adjacent portion of the molecule through one of the methylene (-CH 2 groups present on Y moiety.
15 The structures of a series of novel derivatized haptens which are preferred for use in the invention are shown in Table I. The abbreviations set forth in Table I will be used later to refer to these novel haptens.
a. a 6* 0
S
*I 00 4 LL -I 1
I
I
V--
H- 7586 -16- TABLE I DERIVATIZED HAPTENS OF THE INVENTION p-R2 -NH-C-NH-C 6
H
4
-CH
2
-EDTA-M
11
S
S.
U.
S S
S.
S
S.
*0 0 *0 S
S
S*
S
S S @0 0 55 5 0 0@
-H
-CH
2
CH
3
-(CH
2 )aCH 3
-(CH
2 7
CH
3
-CH
2
'OH
2
OH
-CH
2
OH
2
NH
2
-C
6
H
5
-CH
2
C
6
H
5 -4-(C 6
H
4
)CH
2 C0 2
H
(CH
2 2 00 2
H
-(CH
2 3 C0 2
H
-(CH
2 4
CH(NHCOCH
3
)CONH
2
(CH
2 2 NHCO (CH 2 3 C0 2
H
-C
6
H
4
-CH
2
-DTPA
-CH
2
CH
2 -NHC(=S )NH-C 6
H
4
-CH
2
DTPA
-(CH
2 4
-NHC(=S)NH-C
6
H
4
-CH
2
DTPA
HAPTEN
ABBREVIATION
TUBE
ETUBE
BUTUBE
OTUBE
EOTUBE
NUBE
PHTUBE
BETUBE
PHATUBE
PATUBE
13ATUBE
BALTUBE
GNUBEI
DB-III
DB-IV
DB-V
00 0 9 0* 5* Of* S 9 05 5* Additionally preferred compounds\c~ the invention are those provided in Table II: 6 H-7586 -17- Table II
X-CH
2
-(C=O)-NH-C
6
H
4
-CH
2
-EDTA-M
X
ABBREVIATION
-S(CH
2 4
SCH
2
(C=O)NHC
6
H
4
CH
2 DTPA DB-I
-SCH
2
CH(OH)CH(OH)CH
2
SCH
2
(C=O)NHC
6
H
4
CH
2 DTPA DB-II
-NH-C
6
H
4
-CH
2 -DTPA
DB-VI
-NHCH
2
CH(OH)CH
2
NHCH
2
(C=O)NHC
6
H
4
CH
2 DTPA DB-VIII *6 99 0 9.
9 0e 9 699 6 6690 99 9. 0 90 S 9.
II as 9999 9* 9. 1, *9 6 9 .9 *9 S 39 a 6S 9 ,9 9.
Those skilled in the art will recognize that because the pharmacokinetic criteria for imaging and therapeutic agents may differ, the series of novel u se"\ N derivatized haptensA4peifd~be the invention permits the selection of haptens having desirable pharmacokinetics for a particular application of the invention.
For example, a particular derivatized hapten may be desirable as a therapeutic agent because of its ability 20 to accumulate at a disease site but may be undesirable as an imaging agent because of its inability to clear rapidly from circulation. Additionally, those skilled in the art will appreciate that the present invention is not limited to the'specific derivatized haptens set forth in Tables I or II. The present invention contemplates, therefore, modifications to the structures of the novel derivatized haptens specifically disclosed, provided such structural modifications are intended to modify the inherent pharmacokinetic properties of these haptens for purposes of imaging or therapy.
jXjii Y
K
I
H-7586 -18- In accordance with the invention, the derivatization of a hapten to modify, the dissociation rate of the hapten-antibody complex, and the interaction of the hapten with serum and tissue components, can be accomplished by standard procedures well-known in the art.
For example, hapten structure can be modified by derivatization of the aromatic amine in (S)-4-aminobenzyl EDTA by acylation, alkylation, Schiff's base formation (which may or may not be followed by reduction to the amine), conversion to a reactive isocyanate and subsequent conversion to a substituted urea conversion to a sulfonamide or diazotization and subsequent reaction with a nucleophile. These reactions can be acdomplished with compounds containing simple aliphatic chains, or branched chains, or aromatic or heterocyclic compounds which may or may not contain functional groups. (,ee, March, J. Advanced Organic Chemistry, John Wiley Sons, New York, (1985)). Additionally, in accordance 20 with the invention, the structure of a hapten is preferably modified to enhance the biodistribution and localization of the hapten for a particular application Sse, of the invention. Further, it will be understood that haptens derivatized in accordance with the present invention retain the ability to bind the anti-hapten antibody selected for use. s i The preferred haptens~of the invention represented by Formulae and (Ib) are prepared using standard synthetic methods familiar to one skilled in the art. For example, the preferred thiourea-type i' if 1 1 p 1 H-7586 -19compounds of the invention are prepared by reacting a precursor isothiocyanate compound with an amine. The most useful isothiocyanates for purposes of the present invention are isothiocyanatobenzyl EDTA ("ITCBE") and isothiocyanatobenzyl DTPA ("ITCBD"), both of which are prepared according to the teachings of U.S. Patent No.
4,622,420, incorporated herein by reference. Thus, one skilled in the art determines which compound of the *,00 invention to prepare and then reacts one, or possibly 10 both, of ITCBE or ITCBD with the necessary amine compound(s) to obtain the desired structure. Likewise, to prepare the corresponding macrocyclic polyamine derivatives of the invention, the corresponding isothiocyanate derivatives of DOTA, HETA, TRITA, and TETA are described in U.S. Pat. NO. 4,678,667, the teachings of which are incorporated herein by reference. The prepar- S' ,ation of HETA, DOTA, TRITA and TETA also are disclosed in U.S. Pat. No. 4,678,667. An improved synthesis of DOTA is described in Moi, et al., J. Am. Chem. Soc., 20 110, 6266 (1988). The corresponding isothiocyanate compounds useful as starting materials in the present invention are prepared in a manner completely analogous !to the preparation of ITCBD and ITCBE.
To prepare the amide-type compounds of the invention, two starting materials are particularly useful: -bromoacetamidobenzyl EDTA ("BABE") and S-bromoacetamidobenzyl DTPA These compounds are prepared according to the teachings of DeReimer, et al., J. Lab. Comp. and Radiopharm., 18, 1517 (1981) and U.S. Patent No. 4,622,420. The corresponding LL_' _1 H-7586 bromoacetamido-derivatives for HETA, DOTA, TRITA, and TETA are prepared in a manner completely analogous to the preparation of BABE and BABD. The preparation of these starting materials is described in U.S. Pat. No.
4,678,667, incorporated herein by reference. The alpha-bromo moiety of these compounds renders the adjacent methylene moiety particularly susceptible to nucleophilic attack from groups such as free sulfhydryl or amino groups. Thus, to prepare the desired compound of the invention, one needs only to determine the appropriate intermediate R 2 moiety of Formulae or (Ib) and react it with the required starting material(s).
Typically, the desired compounds of the invention are prepared in aqueous buffer solutions at pH levels from about 6 to about 12. Preferred S* pH ranges are from about 8 to about 11. The progress of the reaction can be monitored easily using HPLC. The temperature is not crucial but prefer- 20 ably the reaction is run at temperatures from about 00 C. to about 600 C, preferably from about 200 C. to about 400 C.
The product obtained from the reaction can be purified using standard chromatography techniques. The preferred separation method involves anion-exchange chromatography using, for example, a DEAE Sephadex resin (Pharmacia Fine Chemicals) or an AG-1X8, formate column (BioRad Laboratories, Richmond California). Of course, those skilled in the art will recognize other '-i 4308a/jj -21equally useful means for purification of the final products.
In addition, It will be recognized that many compounds useful in the present invention contain basic and acidic groups which are capable of forming pharmaceutically-acceptable acid or base addition salts. For example, those compounds which contain a free carboxyl group are able to react with alkaline earth metal bases, for example, sodium bicarbonate, sodium hydroxide, potassium carbonate, potassium bicarbonate, potassium hydroxide, calcium hydroxide, etc., to form pharmaceutically-acceptable salts. Likewise, free amino groups present in the molecule are capable of reacting with acidic reagents to form the Scorresponding acid addition salts. For example, an aminocontaining compound of the invention could be reacted with acids such as hydrochloric, sulfuric, tartaric, lactic, nitric, etc., S: c to prepare the correspondi.ng salt. "Pharmaceuticallyacceptable" in this context refers to those salt forms which are useful in the treatment or diagnosis of a warm-blooded animal.
Although the methods described here and further elaborated In the accompanying non-limiting examples are the presently preferred method for preparing the compounds of the invention, *one skilled in the art will recognize other means which may work equally as well. Thus, the methods provided for preparing com- 1 1.
t -4, 4308a/i -22pourds of Formulae or JIb) are not to be construed as limiting on the means for prepariog the desired compound. Other equal ly- ceptable means for preparing the %.ompounds may be available and are meant to be encompassed by the present invention.
Thus, in another aspect of the invention there is provided a process for preparing a compound of Formula p-R 1 -C 6 H 4 -CR 2 EDTA-M (ID .5S 0 wherein R is -NH-C-NH-R 2
-NH-C-NH-R
2 H-C-C-R NHCCHNH 0 0 OR -NH-C-CH 2
-S-R
2 2 2- 00:0 unsubstituted C 1 to C 30 branched or straight *0 S chain alkyl group; a substituted C 1 to C 30 .0 0 0branched or straight chain alkyl cycloalkyl aryl or arylalkyl group, in which the substituents are one or more of any of: hydroxy, carboxy, -C=O,
SR
fluoro, chloro, bromo, iodo, amino, nitro, -SO 3 H, -NHR 3 -NHR 4
-N(R
3 2
-CONHR
3 -COOR 3 so 4 -PO0 4 phenyl, benzyl, imidazolo, or a group of the formulae: W '308a/jj H- 7586 -23-
-NH-C-NH-R
3
-NH-C-NH-R
3 or -NH-C-NHi-R 3 NH 0 S wherein
R
3 is hydrogen, a C 1 to C 3 0 straight or branched chain alkyl group, -p-phenyl-CH 2
-Y,
-C-CH
3 B e 9a I IG 0 or isoynbzl EDTA, DTPA T, EATapopriteA r 4 iHor-H--R;anMi a metal ionethrof it whic comprisesS R2NCS sarpito (A)inthe cease inwhof w 1 ih aNHC=SnH-R 2 o th. forula
