Title:  Methods of treating autoimmune diseases with a CD86-specific immunotoxin

United States Patent:  6,346,248

Assignee:  Innogenetics N.V. (NL)

Filed:  August 12, 1999

Foreign Application Priority Data:  Jun 07, 1995[EP] (95870066)

A method for treating autoimmune diseases comprising administering to a patient in need of such treatment a therapeutically effective amount of an immunotoxin comprising an anti-human CD86 monoclonal antibody IG10H6D10 as deposited in the ECACC collection under No. 95060210 or a humanized antibody, a single-chain antibody or fragments and specificity of said monoclonal antibody, coupled to a toxin or active fragments thereof wherein the binding of the immunotoxin to CD86 results in the killing of the CD86 expressing cell.

SUMMARY OF THE INVENTION

The current invention is based on the discovery that conjugates of antibodies to human CD80 or human CD86 and a toxin can effectively kill cells expressing these molecules. Accordingly, these antibody-toxin conjugates (which are referred to as "immunotoxins" throughout the remainder of the invention) can be used to prevent or treat (terms used interchangedly) diseases or conditions that are directly or indirectly mediated by the target molecules of the immunotoxin or by the cells carrying the target molecules.

More particularly, the present invention relates to an immunotoxin molecule comprising an antibody specific for human CD80 or CD86 antigen located on the surface of a human cell, coupled to a toxin molecule or active fragment thereof, wherein the binding of the immunotoxin to the CD80 or CD86 antigen results in the killing of the CD80 or CD86 expressing cell.

More particularly, the present invention relates to an immunotoxin molecule comprising an antibody specific for the human CD80 antigen located on the surface of a human cell, coupled to a toxin molecule or active fragment thereof, wherein the binding of the immunotoxin to the CD80 antigen results in the killing of the CD80 expressing cell.

More particularly, the present invention relates to an immunotoxin molecule comprising an antibody specific for the human CD86 antigen located on the surface of a human cell, coupled to a toxin molecule or active fragment thereof, wherein the binding of the immunotoxin to the CD86 antigen results in the killing of the CD86 expressing cell.

According to a possible embodiment, the present invention relates to an immunotoxin comprising an antibody specific for human CD80 and human CD86 both located on the surface of a human cell, coupled to a toxin molecule or active fragment thereof, wherein the binding of the immunotoxin to the CD86 and CD80 antigen results in the killing of the CD80 and CD86 expressing cell.

According to a preferred embodiment, the invention provides an immunotoxin molecule as defined above, wherein said toxin is saporin, gelonin, Pseudomonas exotoxin or Pokeweed antiviral protein or an active fragment thereof.

According to a most preferred embodiment, the invention provides an immunotoxin molecule as defined above, wherein said toxin is gelonin or an active fragment thereof.

According to a preferred embodiment, the invention provides an immunotoxin as defined above, wherein said antibody is a monoclonal antibody or preferably a humanized antibody or a single-chain antibody or a fragment derived from said monoclonal antibody which all have largely retained the specificity of said monoclonal antibody.

According to a preferred embodiment, the present invention relates to immunotoxin molecule as defined above, wherein said monoclonal antibody is the anti-human CD80 monoclonal antibody 5B5D1 as deposited in the ECACC (European Collection of Cell Cultures) collection under No. 95060211 on Jun. 2, 1995 or a humanized antibody, a single-chain antibody or fragments thereof which have largely retained the specificity of said monoclonal antibody.

According to an even preferred embodiment, the present invention provides an immunotoxin molecule as defined above, wherein said monoclonal antibody is the anti-human CD86 monoclonal antibody 1G10H6D10 as deposited in the ECACC collection under No. 95060210 on Jun. 2, 1995 or a humanized antibody, a single-chain antibody or fragments thereof which have largely retained the specificity of said monoclonal antibody.

According to an even more preferred embodiment, the present invention relates to an immunotoxin as defined above, wherein said single chain antibody and said toxin are encoded by a single recombinant DNA (nucleic acid) molecule. This type of recombinant DNA molecule will be named a "recombinant immunotoxin of the invention".

The present invention particularly contemplates recombinant immunotoxin fusion proteins (containing a nucleic acid encoding part of the variable region of the monoclonal antibody fused in frame to a nucleic acid encoding part of a toxin molecule) inspired on the anti-human CD80 monoclonal antibody 5B5D1 as deposited in the ECACC (European Collection of Cell Cultures) collection under No. 95060211 on Jun. 2, 1995 and of the anti-human CD86 monoclonal antibody 1G10H6D10 as deposited in the ECACC collection under No. 95060210 on Jun. 2, 1995.

The present invention also contemplates said recombinant immunotoxin DNA molecule.

The present invention also contemplates recombinant vectors comprising a nucleic acid sequence encoding the amino acid sequence of such a single chain antibody-toxin fusion product (recombinant immunotoxin).

The present invention also contemplates hosts cells transformed by such a vector (and expressing said recombinant immunotoxin).

According to yet another embodiment, the present invention relates to a method for producing an immunotoxin as defined above comprising the steps of chemically or enzymatically coupling a ligand portion consisting of an anti-human CD80 or anti-human CD86 antibody or a derivative thereof as defined above to a toxin portion or a fragment thereof as defined above and isolating the conjugates from the non-conjugated material by any method known in the art.

According to yet another embodiment, the present invention relates to a method for producing a recombinant immunotoxin as defined above comprising the steps of culturing a host cell as defined above which is transformed with a vector comprising a nucleic acid sequence encoding said recombinant immunotoxin under suitable conditions and purifying said immunotoxin by any method known in the art.

According to another embodiment, the present invention provides a composition comprising an immunotoxin molecule as defined above in a pharmaceutically acceptable excipient for use in therapy (this type of compositions is referred to as "composition comprising the same" throughout the remainder of the invention).

According to yet another embodiment, the present invention provides a medicament comprising an immunotoxin molecule as defined above.

According to another embodiment, the present invention provides an immunotoxin molecule or a composition comprising the same as defined above for use as a medicament.

According to still another embodiment, the present invention relates to the use of an immunotoxin molecule or a composition comprising the same for the preparation of a medicament for treating diseases of the immune system, in particular for preventing allograft rejection, treating autoimmune diseases and treating various malignancies of lymphoid origin such as Hodgkin's Disease (the term "treating" is to be understood as referring to the treatment of a patient suffering from a disease as well as to the prophylactic or preventive treatment of an individual liable to suffer from a disease).

According to a particularly preferred embodiment, the present invention relates to a method for preventing allograft transplant rejection, the method comprising administering to a patient in need of such treatment a therapeutically effective amount of an immunotoxin or a composition comprising the same as defined above, wherein the binding of the immunotoxin to the CD80 or CD86 antigen prevents the activation and differentiation of host T-cells against the MHC on the allograft, in a pharmaceutically acceptable excipient.

According to a particularly preferred embodiment, the present invention provides an immunotoxin or a composition comprising the same as defined above for use in a method for preventing allograft transplant rejection, the method comprising administering to a patient in need of such treatment a therapeutically effective amount of such an immunotoxin or such a composition, wherein the binding of the immunotoxin to the CD80 or CD86 antigen prevents the activation and differentiation of host T-cells against the MHC on the allograft, in a pharmaceutically acceptable excipient.

According to a preferred embodiment, the present invention provides for the use of an immunotoxin or a composition comprising the same as defined above for the preparation of a medicament for preventing allograft transplant rejection.

According to a further embodiment, the present invention provides a method for preventing allograft transplant rejection, the method comprising administering to a patient in need of such treatment a therapeutically effective amount of an immunotoxin or a composition comprising the same, wherein the binding of the immunotoxin to the CD80 antigen prevents the activation and differentiation of host T cells against the MHC on the allograft, in a pharmaceutically acceptable excipient.

According to yet another embodiment, the present invention provides a method for preventing allograft transplant rejection, the method comprising administering to a patient in need of such treatment a therapeutically effective amount of an immunotoxin or a composition comprising the same, wherein the binding of the immunotoxin to the CD86 antigen prevents the activation and differentiation of host T cells against the MHC on the allograft, in a pharmaceutically acceptable excipient.

According to another embodiment, the present invention provides a method for preventing allograft transplant rejection, the method comprising ex vivo perfusion of a donor organ with a therapeutically effective amount of an immunotoxin or a composition comprising the same as defined above, wherein the binding of the immunotoxin to the CD80 or CD86 antigen results in the killing of said CD80 or CD86 expressing cells.

According to another embodiment, the present invention provides an immunotoxin or composition comprising the same as defined above for use in a method for preventing allograft transplant rejection, the method comprising ex vivo perfusion of a donor organ with a therapeutically effective amount of an immunotoxin or such a composition, wherein the binding of the immunotoxin to the CD80 or CD86 antigen results in the killing of said CD80 or CD86 expressing cells.

According to another embodiment, the present invention relates to the use of an immunotoxin or a composition comprising the same as defined above for the preparation of medicament for preventing allograft transplant rejection involving ex vivo perfusion of a donor organ with a therapeutically effective amount of said immunotoxin or said composition.

According to yet another embodiment, the present invention relates to a method for preventing allograft transplant rejection, the method comprising ex vivo perfusion of a donor organ with a therapeutically effective amount of an immunotoxin or a composition comprising the same, wherein the binding of the immunotoxin to the CD80 antigen results in killing of said CD80 expressing cells.

According to an alternative embodiment, the present invention relates to a method for preventing allograft transplant rejection, the method comprising ex vivo perfusion of a donor organ with a therapeutically effective amount of an immunotoxin or a composition comprising the same, wherein the binding of the immunotoxin to the CD86 antigen results in the killing of said CD86 expressing cells.

According to another embodiment, the present invention relates to a donor organ which has been ex vivo perfused with a therapeutically effective amount of an immunotoxin or a composition comprising the same.

According to an alternative embodiment, the present invention relates to a method for producing a donor organ as defined above, comprising the step of ex vivo perfusion of said donor organ with a therapeutically effective amount of an immunotoxin or a composition comprising the same.

According to another embodiment, the present invention relates to isolated lymphocytes which have been incubated ex vivo with a therapeutically effective amount of an immunotoxin or a composition comprising the same.

According to yet another embodiment, the present invention relates to a method for producing isolated lymphocytes as defined above, comprising the step of ex vivo incubating said isolated lymphocytes with a therapeutically effective amount of an immunotoxin or a composition comprising the same as defined above.

According to yet another embodiment, the present invention provides a method for induction of alloantigen-specific tolerance, the method comprising administering to a patient in need of such treatment a therapeutically effective amount of lymphocytes from an organ donor that have been incubated ex vivo with a therapeutically effective amount of an immunotoxin or such a composition, wherein the binding of the immunotoxin to the CD80 or CD86 antigen results in the killing of said CD80 or CD86 expressing cells, in a pharmaceutically acceptable excipient.

According to another embodiment, the present invention relates to an immunotoxin or a composition comprising the same as defined above for use in a method for induction of alloantigen-specific tolerance, the method comprising administering to a patient in need of such treatment a therapeutically effective amount of lymphocytes from an organ donor that have been incubated ex vivo with a therapeutically effective amount of an immunotoxin or a composition comprising the same as defined above, wherein the binding of the immunotoxin to the CD80 or CD86 antigen results in the killing of said CD80 or CD86 expressing cells, in a pharmaceutically acceptable excipient.

According to another embodiment, the present invention relates to the use of an immunotoxin or a composition comprising the same for the preparation of a medicament for induction of allogen-specific tolerance.

According to another alternative embodiment, the present invention relates to a method for induction of alloantigen-specific tolerance, the method comprising administering to a patient in need of such treatment a therapeutically effective amount of lymphocytes from an organ donor that have been incubated ex vivo with a therapeutically effective amount of an immunotoxin or a composition comprising the same, wherein the binding of the immunotoxin to the CD80 antigen results in killing of said CD80 expressing cells, in a pharmaceutically acceptable excipient.

It is another objective of this invention to provide a method for induction of alloantigen-specific tolerance, the method comprising administering to a patient in need of such treatment a therapeutically effective amount of lymphocytes from an organ donor that have been incubated ex vivo with a therapeutically effective amount of an immunotoxin or a composition comprising the same, wherein the binding of the immunotoxin to the CD86 antigen results in killing of said CD86 expressing cells, in a pharmaceutically acceptable excipient.

According to yet another embodiment, the present invention relates to a method for preventing or treating autoimmune diseases such as rheumatoid arthritis and systematic lupus erythematosus in a patient, the method comprising administering to a patient in need of such treatment a therapeutically effective amount of an immunotoxin or a composition comprising the same, wherein the binding of the immunotoxin to the CD80 or CD86 antigen results in elimination of the CD80 or CD86 expressing cells and inhibition of the local inflammatory response, in a pharmaceutically acceptable excipient.

According to yet another embodiment, the present invention relates to an immunotoxin or a composition comprising the same as defined above for use in a method for preventing or treating autoimmune diseases such as rheumatoid arthritis and systematic lupus erythematosus in a patient, the method comprising administering to a patient in need of such treatment a therapeutically effective amount of an immunotoxin or a composition comprising the same, wherein the binding of the immunotoxin to the CD80 or CD86 antigen results in elimination of the CD80 or CD86 expressing cells and inhibition of the local inflammatory response, in a pharmaceutically acceptable excipient.

According to yet another embodiment, the present invention relates to the use of an immunotoxin or a composition comprising the same as defined above for the preparation of a medicament for preventing or treating autoimmune diseases such as rheumatoid arthritis and systematic lupus erythematosus in a patient.

According to yet another embodiment, the present invention relates to a method for preventing or treating autoimmune diseases such as rheumatoid arthritis and systematic lupus erythematosus in a patient, the method comprising administering to a patient in need of such treatment a therapeutically effective amount of an immunotoxin or a composition comprising the same, wherein the binding of the immunotoxin to the CD80 antigen results in elimination of the CD80 expressing cells and, inhibition of the local inflammatory response, in a pharmaceutically acceptable excipient.

According to yet another embodiment, the present invention relates to a method for preventing or treating autoimmune diseases such as rheumatoid arthritis and systematic lupus erythematosus in a patient, the method comprising administering to a patient in need of such treatment a therapeutically effective amount of an immunotoxin or a composition comprising the same, wherein the binding of the immunotoxin to the CD86 antigen results in elimination of the CD86 expressing cells and inhibition of the local inflammatory response, in a pharmaceutically acceptable excipient.

According to another embodiment, the present invention relates to a method for treating malignancies of lymphoid origin such as Hodgkin's disease in a patient, the method comprising administering to a patient in need of such treatment a therapeutically effective amount of lymphocytes from an organ donor that have been incubated ex vivo with a therapeutically effective amount of an immunotoxin or a composition comprising the same, wherein the binding of the immunotoxin to the CD80 or CD86 antigen results in elimination of the CD80 or CD86 expressing tumor cells, in a pharmaceutically acceptable excipient.

According to yet another embodiment, the present invention relates to an immunotoxin or a composition comprising the same for use in a method for treating malignancies of lymphoid origin such as Hodgkin's disease in a patient, the method comprising administering to a patient in need of such treatment a therapeutically effective amount of an immunotoxin or such a composition, wherein the binding of the immunotoxin to the CD80 or CD86 antigen results in elimination of the CD80 or CD86 expressing tumor cells, in a pharmaceutically acceptable excipient.

According to yet another embodiment, the present invention relates to the use of an immunotoxin or a composition comprising the same for the preparation of a medicament for treating malignancies of lymphoid origin such as Hodgkin's disease in a patient.

According to yet another embodiment, the present invention relates to a method for treating malignancies of lymphoid origin in a patient, the method comprising administering to a patient in need of such treatment a therapeutically effective amount of an immunotoxin or a composition comprising the same, wherein the binding of the immunotoxin to the CD80 antigen results in elimination of the CD80 expressing tumor cells, in a pharmaceutically acceptable excipient.

According to yet another embodiment, the present invention relates to a method for treating malignancies of lymphoid origin in a patient, the method comprising administering to a patient in need of such treatment a therapeutically effective amount of an immunotoxin or a composition comprising the same, wherein the binding of the immunotoxin to the CD86 antigen results in elimination of the CD86 expressing tumor cells, in a pharmaceutically acceptable excipient.

According to a particularly preferred embodiment, the present invention relates to a method for treating Hodgkin's disease in a patient, the method comprising administering to a patient in need of such treatment a therapeutically effective amount of an immunotoxin or a composition comprising the same, wherein the binding of the immunotoxin to the CD80 antigen results in elimination of the CD80 expressing tumor cells, in a pharmaceutically acceptable excipient.

According to a particularly preferred embodiment, the present invention relates to a method for treating Hodgkin's disease in a patient, the method comprising administering to a patient in need of such treatment a therapeutically effective amount of an immunotoxin or a composition comprising the same, wherein the binding of the immunotoxin to the CD86 antigen results in elimination of the CD86 expressing tumor cells, in a pharmaceutically acceptable excipient.

Claim 1 of 4 Claims

What is claimed is:

1. A method for treating autoimmune diseases comprising administering to a patient in need of such treatment a therapeutically effective amount of an immunotoxin comprising an anti-human CD86 monoclonal antibody IG10H6D10 as deposited in the ECACC collection under No. 95060210 or a humanized antibody, a single-chain antibody or fragments and specificity of said monoclonal antibody, coupled to a toxin or active fragments thereof wherein the binding of the immunotoxin to CD86 results in the killing of the CD86 expressing cell.

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