Modified antibodies of induced anti-idiotype reaction enhancement

Abstract

The present invention relates to the modification of immunoglobulin molecules in which one or more cysteine ​​residues in the variable region which form intrachain disulfide bonds are replaced by amino acids which do not contain a sulfhydryl group and thus do not form disulfide bonds. The ability of these modified immunoglobulin molecules to induce an anti-idiotypic response is enhanced. The present invention further provides a method for treating or preventing tumors and / or infectious diseases using the modified immunoglobulin of the present invention.

Description

[0001] Cross-references to related applications:

[0002] This application claims the benefit of two applications, Provisional Application No. 60 / 065,716, filed November 14, 1997, and Provisional Application No. 60 / 081,403, filed April 10, 1998, which are incorporated herein in their entirety Incorporated by reference. 1. Field of invention:

[0003] The present invention relates to modified immunoglobulins and vaccine compositions thereof, wherein one or more cysteine ​​residues in the variable region that form an intrachain disulfide bond are replaced with an amino acid that does not contain a sulfhydryl group, thereby not forming disulfide bond. The invention also relates to the use of the vaccine compositions of the invention to treat or prevent certain diseases and disorders, especially cancer and infectious diseases. 2. Background of the invention

[0004] 2.1 Immunoglobulin structure

[0005] The basic unit of immunoglobulin structure is a complex of four polypepti...

Examples

preparation example Construction

[0116] Alternatively, disulfide-forming cysteines may be replaced with non-sulfhydryl-containing atypical amino acids or chemical amino acid analogs (such as, but not limited to, conventional protein synthesis methods). Atypical amino acids include: D-amino acids of typical amino acids, α-aminobutyric acid, 4-aminobutyric acid, Abu, 2-aminobutyric acid, γ-Abu, ε-Ahx, -aminocaproic acid, Aib, 2-aminoisobutyric acid Butyric acid, 3-aminopropionic acid, ornithine, norleucine, norvaline, hydroxyproline, sarcosine, citrulline, tert-butylglycine, tert-butylalanine, phenyl Glycine, cyclohexylalanine, β-alanine, fluoro-amino acids such as designed β-methyl amino acids, Cα-methyl amino acids, Nα-methyl amino acids, amino acid analogs. Also, amino acids can be right-handed or left-handed. In alternative embodiments, disulfide bond forming amino acid residues may be deleted.

[0117] In specific embodiments, disulfide bond-forming amino acid residues in the variable region of the heavy...

Embodiment 7

[0364] Example 7 Expression of Synthetic Modified Antibody Containing HMFG-1 Sequence

[0365]Anti-idiotypic antibodies that immunospecifically bind to HMFG-1 antibodies were constructed. HMFG-1 is an antibody known to bind polymorphic epidermal mucin (PEM) (Stewart et al., 1990, Journal of Clinical Oncology 8: 1941-50; Kosmas et al., 1994, Cancer 73: 3000-3010 ). The antigenic determinant sequence ProAspThrArgPro of PEM is inserted into the variable region of the antibody by the method of the present invention. This short sequence is a highly immunogenic region in human polymorphic epidermal mucin (Gendler et al., 198, J. Biol. Chem. 163: 12820-12823). The 27A-27E (SerLeuLeuTyrSer) residues of HMFG-1 (Table 6) were replaced with ProAspThrArgPro by the oligonucleotide synthesis method described in Section 6, see FIG. 10 . Using the known gene sequence of the light and heavy chain variable regions of the HMFG-1 antibody, an anti-idiotypic antibody that immunospecifically bin...

[0001] Cross-references to related applications:

[0002] This application claims the benefit of two applications, Provisional Application No. 60 / 065,716, filed November 14, 1997, and Provisional Application No. 60 / 081,403, filed April 10, 1998, which are incorporated herein in their entirety Incorporated by reference. 1. Field of invention:

[0003] The present invention relates to modified immunoglobulins and vaccine compositions thereof, wherein one or more cysteine ​​residues in the variable region that form an intrachain disulfide bond are replaced with an amino acid that does not contain a sulfhydryl group, thereby not forming disulfide bond. The invention also relates to the use of the vaccine compositions of the invention to treat or prevent certain diseases and disorders, especially cancer and infectious diseases. 2. Background of the invention

[0004] 2.1 Immunoglobulin structure

[0005] The basic unit of immunoglobulin structure is a complex of four polypepti...

Examples

preparation example Construction

[0116] Alternatively, disulfide-forming cysteines may be replaced with non-sulfhydryl-containing atypical amino acids or chemical amino acid analogs (such as, but not limited to, conventional protein synthesis methods). Atypical amino acids include: D-amino acids of typical amino acids, α-aminobutyric acid, 4-aminobutyric acid, Abu, 2-aminobutyric acid, γ-Abu, ε-Ahx, -aminocaproic acid, Aib, 2-aminoisobutyric acid Butyric acid, 3-aminopropionic acid, ornithine, norleucine, norvaline, hydroxyproline, sarcosine, citrulline, tert-butylglycine, tert-butylalanine, phenyl Glycine, cyclohexylalanine, β-alanine, fluoro-amino acids such as designed β-methyl amino acids, Cα-methyl amino acids, Nα-methyl amino acids, amino acid analogs. Also, amino acids can be right-handed or left-handed. In alternative embodiments, disulfide bond forming amino acid residues may be deleted.

[0117] In specific embodiments, disulfide bond-forming amino acid residues in the variable region of the heavy...

Embodiment 7

[0364] Example 7 Expression of Synthetic Modified Antibody Containing HMFG-1 Sequence

[0365]Anti-idiotypic antibodies that immunospecifically bind to HMFG-1 antibodies were constructed. HMFG-1 is an antibody known to bind polymorphic epidermal mucin (PEM) (Stewart et al., 1990, Journal of Clinical Oncology 8: 1941-50; Kosmas et al., 1994, Cancer 73: 3000-3010 ). The antigenic determinant sequence ProAspThrArgPro of PEM is inserted into the variable region of the antibody by the method of the present invention. This short sequence is a highly immunogenic region in human polymorphic epidermal mucin (Gendler et al., 198, J. Biol. Chem. 163: 12820-12823). The 27A-27E (SerLeuLeuTyrSer) residues of HMFG-1 (Table 6) were replaced with ProAspThrArgPro by the oligonucleotide synthesis method described in Section 6, see FIG. 10 . Using the known gene sequence of the light and heavy chain variable regions of the HMFG-1 antibody, an anti-idiotypic antibody that immunospecifically bin...

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