Trifunctional antibody-like compounds as a combined diagnostic and therapeutic agent

• US 5,273,743 A
• Filed: 12/13/1991
• Issued: 12/28/1993
• Est. Priority Date: 03/09/1990
• Status: Expired due to Fees

• Current Assignee: Hybritech, Inc. (Danaher Corporation)
• Original Assignee: Hybritech, Inc. (Danaher Corporation)
• Inventors: Ahlem, Clarence, Huang, Ann E.
• Primary Examiner(s): KIM, KAY KWANG AE
• Application Number:
  US07/808,193
• Time in Patent Office: 746 Days
• Field of Search: 424/85.8, 424/85.91, 424/1.1, 424/9, 530/387.3, 530/388.2, 530/389.7, 530/391.1, 530/391.3, 530/391.5, 530/391.7, 530/391.9, 530/388.85, 530/388.9, 530/389.8, 436/819, 435/972
• US Class Current: 424/136.1
• CPC Class Codes: A61K 2123/00 Preparations for testing in vivo
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 47/6879 the immunoglobulin having two or more different antigen-binding sites, e.g. bispecific or multispecific immunoglobulin
• CPC Class Codes: A61K 49/16 Antibodies; Immunoglobulins; Fragments thereof
• CPC Class Codes: A61K 51/1093 conjugates with carriers being antibodies
• CPC Class Codes: C07K 16/468 Immunoglobulins having two or more different antigen binding sites, e.g. multifunctional antibodies
• CPC Class Codes: Y10S 435/972 Modified antibody, e.g. hybrid, bifunctional
• CPC Class Codes: Y10S 436/819 Multifunctional antigen or antibody

Abstract

The present invention has multiple aspects. In its first aspect the present invention is directed to a trifunctional antibody-like compound that has tissue, organ, cell or molecule specificity and which is capable of being bifunctional when immobilized, via binding, at the tissue, organ, cell or molecule for which it has specificity. In particular, the present invention is directed to a trifunctional antibody-like compound of Formula I:

F.sub.1 ab'"'"'--L--F.sub.2 ab'"'"'--L--F.sub.3 ab'"'"'                (1)

wherein L is the same or two different moieties for cross-linking F₁ ab'"'"', F₂ ab'"'"' and F₃ ab'"'"';

wherein F₁ ab'"'"' is an Fab'"'"'-like fragment of a polyclonal or monoclonal antibody having specificity for an antigen expressed by the organ, tissue, cell or molecule of interest;

wherein F₂ ab'"'"' is an Fab'"'"'-like fragment of a polyclonal or monoclonal antibody having the same specificity as F₁ ab'"'"', or having specificity either for a different antigen expressed by the organ, tissue, cell, or molecule of interest, or for a diagnostic or therapeutic agent; and

wherein F₃ ab'"'"' is an Fab'"'"'-like fragment of a polyclonal or monoclonal antibody having specificity for a diagnostic or therapeutic agent; or

wherein F₂ ab'"'"' and F₃ ab'"'"' have specificity for a receptor/receptor complex and accessory molecule on the surface of a T-cell.

The compound of the present invention has utility as a pharmaceutical agent. Various methods for treating and diagnosing a disease are also presented.

Claims

1. A pharmaceutical composition comprising:

(a) a compound of the formula;

F₁ ab'"'"'--L--F₂ ab'"'"'--L--F₃ ab'"'"' wherein F₁ ab'"'"', F₂ ab'"'"', and F₃ ab'"'"' are three different Fab'"'"'-like fragments and wherein L is a bis-succinimidyl and/or a tris-succinimidyl moiety that is the same or different, each L covalently cross-linking two of said three different Fab'"'"'-like fragments, said three different and cross-linked Fab'"'"'-like fragments substantially retaining their antigen binding activity; and

(b) one or more pharmaceutically acceptable carriers.

2. The pharmaceutical composition of claim 1 wherein any one of F₁ ab'"'"', F₂ ab'"'"' or F₃ ab'"'"' is an Fab'"'"'-like fragment of a monoclonal antibody that is capable of forming a binding pair in vivo with a physiologically compatible chelate complex comprising a chelating agent and a beta emitting metal ion.

3. The pharmaceutical composition of claim 2 wherein said physiologically compatible chelate complex has a dissociation constant of 10^-16 to 10^-20.

4. The pharmaceutical composition of claim 3 wherein the chelating agent is selected from the group consisting of ethylenediaminetetraacetic acid ("EDTA"), ethanolaminethioureabenzyl-EDTA ("EOTUBE"), diethylenetriaminepentaacetic acid ("DTPA"), methylthioureabenzyl DTPA ("MeTUBD"), 1,4,7,10-tetrazacyclododecane-N'"'"',N",N'"'"'",N""-tetraacetic acid ("DOTA"), L-aminobenzyl-EDTA, 1,5,9,13-tetraazacyclohexadecane-N, N'"'"', N", N'"'"'"-tetraacetic acid ("HETA"), 1,4,7,10-tetraazacyclotridecane-N, N'"'"', N", N'"'"'"-tetraacetic acid, and 1,4,8,11-tetraazacyclotetradecane-N, N'"'"', N", N'"'"'"-tetraacetic acid ("TETA").

5. The pharmaceutical composition of claim 4 wherein the beta emitting metal ion is selected from the group consisting of ⁶⁷ Cu, ¹⁸⁶ Rh, ¹⁸⁸ Rh, ¹⁸⁹ Rh, ¹⁵³ Sm and ⁹⁰ Y, in ionic form.

6. The pharmaceutical composition of claim 5 wherein the physiologically compatible chelate complex is ⁹⁰ Y-MeTUBD, ⁹⁰ Y-DTPA, or ⁹⁰ Y-DOTA.

7. The pharmaceutical composition of claim 6 wherein each L is a trissuccinimidyl moiety.

8. The pharmaceutical composition of claim 7 wherein a second one of F₁ ab'"'"', F₂ ab'"'"' or F₃ ab'"'"' is capable of forming a binding pair in vivo with a second physiologically compatible chelate complex, said second physiologically compatible complex comprising a second chelating agent and a gamma emitting metal ion.

9. The pharmaceutical composition of claim 8 wherein said second chelating agent is selected from the group consisting of EDTA, EOTUBE, DTPA, MeTUBD, DOTA, HETA, TRITA and TETA.

10. The pharmaceutical composition of claim 9 wherein the gamma emitting metal ion is selected from the group consisting of ¹¹¹ In, ^113m In, ⁶⁷ Ga, ⁶⁸ Ga, ^99m Tc, ⁵¹ Cr, ¹⁹⁷ Hg, ²⁰³ Hg, ¹⁶⁹ Yb, ⁸⁵ Sr., and ⁸⁷ Sr, in ionic form.

11. The pharmaceutical composition of claim 10 wherein a third one of F₁ ab'"'"', F₂ ab'"'"' or F₃ ab'"'"' has specificity for an antigen expressed by a cell.

12. The pharmaceutical composition of claim 11 in unit dosage form.

13. A composition comprising a compound of the formula:

space="preserve" listing-type="equation">F.sub.1 ab'"'"'--L--F.sub.2 ab'"'"'--L--F.sub.3 ab'"'"' (I)

wherein F₁ ab'"'"', F₂ ab'"'"' and F₃ ab'"'"' are three different Fab'"'"'-like fragments; and

wherein L is a bis-succinimidyl and/or a tris-succinimidyl moiety that is the same or different, each L covalently cross-linking two of said three different and cross-linked Fab'"'"'-like fragments, said three different Fab'"'"'-like fragments substantially retaining their antigen binding activity;

whereby the compound of Formula I is capable of forming binding pairs with three different epitopes.

14. The composition of claim 13 wherein each L is a tris-succinimidyl moiety.

15. The composition of claim 14 wherein one of said three different Fab'"'"'-like fragments is capable of forming a binding pair with a first physiologically compatible chelate complex comprising a first chelating agent and a first metal ion, said first physiologically compatible chelate complex being characterized by having a dissociation constant of 10^-16 to 10^-20.

16. The composition of claim 15 wherein the first chelating agent is selected from the group consisting of EDTA, DTPA, MeTUBD, DOTA, HETA, TRITA, and TETA.

17. The composition of claim 16 wherein the first metal ion is selected from the group consisting of ¹¹¹ In, ^113m In, ⁶⁷ Ga, ⁶⁸ Ga, ^99m Tc, ⁵¹ Cr, ¹⁹⁷ Hg, ²⁰³ Hg, ¹⁶⁹ Yb, ⁸⁵ Sr, and ⁸⁷ Sr, in ionic form.

18. The composition of claim 17 wherein the first metal ion is ¹¹¹ In(III).

19. The composition of claim 18, wherein a second one of said three different Fab'"'"'-like fragments is capable of forming a binding pair with a second physiological compatible chelate complex comprising, a second chelating agent and a second metal ion, said second physiologically compatible chelate complex being characterized by having a dissociation constant of 10^-16 to 10^-20.

20. The composition of claim 19 wherein the second chelating agent is selected from the group consisting of EDTA, EQTUBE, DTPA, MeTUBD, DOTA, HETA, TRITA and TETA.

21. The composition of claim 20 wherein the second metal ion is selected from the group consisting of ⁶⁷ Cu, ¹⁸⁶ Rh, ¹⁸⁸ Rh, ¹⁸⁹ Rh, ¹⁵³ Sm and ⁹⁰ Y, in ionic form.

22. The composition of claim 21 wherein the second metal ion is ⁹⁰ Y in ionic form and the second chelating agent is DTPA or MeTUBD.

23. The composition of claim 22 wherein a third one of said three different Fab'"'"'-like fragments is capable of binding to an antigen expressed by a cell.

24. The composition of claim 23 wherein each of F₁ ab'"'"', and F₂ ab'"'"', and F₃ ab'"'"' is an Fab'"'"'-like fragment from a monoclonal antibody.

25. The composition of claim 24 wherein the antigen expressed by a cell is selected from the group consisting of alphafetoprotein, c-erbB-2, cancer antigen 15-3 ("CA 15-3"), CA 19-9, CA 125, CA 195, CA 549, carcinoembryonic antigen ("CEA"), cathepsin D ("cath D"), cytokeratins, Du-PAN-2, Epidermal Growth Factor Receptor ("EGF-R"), estrogen receptor, C-myc, N-myc, prostate specific antigen (PSA), ras, tumor associated antigen-72 ("TAG-72"), tumor associated antigen-4 ("TA-4") and the KS 1/4 antigen.

26. The composition of claim 25 wherein the cellular antigen is CEA.