Methods of administering an N-terminal modified PEG-TRAIL
Inventors
Lee, Kang Choon • Lee, Seulki • PARK, Eun Ji
Assignees
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Publication Number
US-10046059-B2
Publication Date
2018-08-14
Expiration Date
Abstract
Disclosed herein are an N-terminal modified PEG-TRAIL conjugates and methods of making and using thereof. The PEG-TAIL conjugates have bioactivity that is substantially similar to that of native TRAIL coupled with an extended in vivo half-life and enhanced stability. The disclosed PEG-TRAIL conjugates exhibit significantly reduced hepatotoxicity when compared to that of non-PEGylated trimeric TRAIL. The disclosed methods of making the PEG-TRAIL conjugates provide a homogeneous, highly pure, form of N-terminal modified PEG-TRAIL. Compared to native TRAIL, the PEG-TRAIL conjugates exhibits high solubility and solution stability. The PEG-TRAIL conjugates are useful in preventing and treating proliferative or autoimmune diseases.
Core Innovation
The invention provides N-terminal modified PEG-TRAIL conjugates in which a trimeric TRAIL comprises three TRAIL monomers. The monomers include either full length human TRAIL or truncated human TRAIL comprising amino acids 114-281 of full length human TRAIL, and each monomer has an N-terminus comprising a zipper amino acid motif. PEG or a PEG derivative is bound to the N-terminus of one of the monomers, promoting N-terminal trimerization through the zipper motif.
The N-terminal modified PEG-TRAIL is purified by ion exchange chromatography to remove any TRAIL monomers or PEG that is not bound to the trimeric PEG-TRAIL. The disclosure includes support for purification to a homogeneous mono-PEGylated trimeric PEG-TRAIL, including removal of unbound components. The PEGylation concept is presented as controlled site-specific attachment at the N-terminus via zipper motifs rather than random PEGylation.
The disclosure states that PEGylation extends in vivo half-life and improves solubility and solution stability. It also states that PEGylation reduces hepatotoxicity while preserving or expanding anti-tumor activity versus native TRAIL and versus randomly PEGylated TRAIL. Documented examples include enhanced potency and improved pharmacokinetics, together with reduced hepatocyte toxicity.
Claims Coverage
The independent claim covers a disease-treatment method using a purified N-terminally PEGylated trimeric PEG-TRAIL, with inventive features including trimeric TRAIL monomer definition, N-terminal zipper-mediated PEG attachment, and ion-exchange purification to remove unbound components. Dependent claims refine PEG chemistry, specify particular TRAIL sequences, narrow cancer indications, constrain dosing frequency, and add comparative functional properties related to protease-mediated degradation.
Purified N-terminally PEGylated trimeric PEG-TRAIL for treating disease
Administering a pharmaceutically effective amount of a PEG-TRAIL consisting of a trimeric TRAIL comprising three TRAIL monomers each having the sequence of full length human TRAIL or truncated human TRAIL comprising amino acids 114-281, wherein the N-terminus of each monomer comprises a zipper amino acid motif and having a PEG or PEG derivative bound to the N-terminus of one of the monomers.
Ion exchange purification to remove unbound components
Purifying the N-terminal modified PEG-TRAIL by ion exchange chromatography to remove any TRAIL monomers or PEG that is not bound to the trimeric TRAIL.
Disease treatment for cancer, immune or inflammatory disorder, or diabetes
Treating a subject in need thereof having cancer, an immune or inflammatory disorder, or diabetes.
Defined PEG derivative chemistries for N-terminal PEG attachment
Wherein the PEG derivative is selected from methoxypolyethylene glycol succinimidyl propionate, methoxypolyethylene glycol N-hydroxysuccinimide, methoxypolyethylene glycol aldehyde, methoxypolyethylene glycol maleimide, or multiple-branched polyethylene glycol.
Specific TRAIL monomer sequences (full-length or truncated) identified by SEQ IDs
Using TRAIL monomers having an amino acid sequence of SEQ ID NO:1 or SEQ ID NO:2.
Specific cancer indications
Treating cancer selected from colon carcinoma, glioma, lung carcinoma, prostate carcinoma, brain tumor, or multiple myeloma.
Dosing frequency constraint
Administering the effective amount once every one to two weeks.
Reduced protein degradation by proteases compared with non-PEGylated TRAIL
Using an N-terminal modified PEG-TRAIL having decreased protein degradation by proteases compared with non-PEGylated TRAIL.
Overall, the claims focus on administering a purified N-terminally PEGylated trimeric PEG-TRAIL in which PEG is attached at the monomer N-terminus via a zipper amino acid motif, with ion exchange purification removing unbound TRAIL monomers or PEG. Dependent claims further specify PEG derivative chemistries, constrain TRAIL to particular sequence identifiers, narrow therapeutic indications, constrain dosing frequency, and add comparative protease-degradation benefits.
Stated Advantages
Extended in vivo half-life.
Increased solubility and solution stability.
Reduced hepatotoxicity.
Preserved or expanded anti-tumor activity versus native TRAIL and versus randomly PEGylated TRAIL.
Enhanced potency and improved pharmacokinetics.
Reduced renal excretion.
Decreased protein degradation by proteases compared with non-PEGylated TRAIL.
Documented Applications
Treating cancer, including colon carcinoma, glioma, lung carcinoma, prostate carcinoma, brain tumor, and multiple myeloma.
Treating immune or inflammatory disorders, including lupus (experimental autoimmune encephalomyelitis) and rheumatoid arthritis.
Treating diabetes, including type I diabetes (insulin-dependent diabetes mellitus).
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