Conjugates of an IL-2 moiety and a polymer
Inventors
Bossard, Mary J. • Ali, Cherie F. • Liu, Xiaofeng • Charych, Deborah H. • Wang, Yujun
Assignees
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Publication Number
US-10960079-B2
Publication Date
2021-03-30
Expiration Date
Abstract
Conjugates of an interleukin-2 (“IL-2”) moiety and one or more nonpeptidic, water-soluble polymers are provided. Typically, the non-peptidic, water-soluble polymer is poly(ethylene glycol) or a derivative thereof. Also provided, among other things, are compositions comprising conjugates, methods of making conjugates, methods of administering compositions to an individual, nucleic acid sequences, expression systems, host cells, and methods for preparing IL-moieties.
Core Innovation
The invention relates to interleukin-2 moiety–polymer conjugates in which a polymer moiety is covalently attached to an IL-2 residue. The polymer moiety is a water-soluble nonpeptidic polymer, including poly(ethylene glycol) (PEG), arranged as linear, branched, forked, or multifunctional PEG architectures, and the PEG is covalently attached to an amino group of an interleukin-2 moiety.
The IL-2 moiety is described within a framework including interleukin-2 moieties having amino acid sequences referenced by SEQ ID NO: 1 through SEQ ID NO: 4, as well as precursor versus non-precursor forms, and glycosylated versus unglycosylated variants. The conjugates include releasable linkages and hydrolytically stable linkages, including ester, carbamate, phosphate, imines, acetal, and hydrazone linkage types, as well as amide, carbamate, urea, and urethane linkage types, with spacer/linker moieties and attachment-point considerations such as lysines, the N-terminus, and carbohydrates.
The disclosed subject matter centers on branched PEG attachment to IL-2 through releasable linkage functionality, including branched poly(ethylene glycol) polymers covalently attached via a releasable linkage to the amino group of the IL-2 moiety. The disclosed architectures also include optional PEG end-capping moieties and thiol-targeting mPEG/IL-2 conjugation chemistry, including thiol-selective activated PEG reagents, thiol-reactive activated PEG reagents, N-maleimidyl polymer reagents, and illustrative conjugate structures with PEG branching and covalent attachment to IL-2.
Claims Coverage
The consolidated independent claim coverage centers on administering to an individual an interleukin-2 conjugate having SEQ ID NO:1, with one to seven branched poly(ethylene glycol) polymers covalently attached via a releasable linkage to an amino group of the interleukin-2 moiety. The claims also specify that each branched PEG has a weight average molecular weight of about 20,000 daltons to about 85,000 daltons. Dependent refinements further specify releasable carbamate linkage, absence of interleukin-2 activity prior to in vivo release, narrower PEG arm-count limits or mixtures with an average arm count, and selected cancer indications.
Branched PEG conjugate of SEQ ID NO:1 IL-2 via releasable linkage
A conjugate of an interleukin-2 moiety having the amino acid sequence of SEQ ID NO:1, wherein one to seven branched poly(ethylene glycol) polymers are covalently attached via a releasable linkage to an amino group of the interleukin-2 moiety.
Specified branched PEG molecular weight range
Each branched poly(ethylene glycol) molecule has a weight average molecular weight from about 20,000 daltons to about 85,000 daltons.
Releasable carbamate linkage
The releasable linkage is a releasable carbamate linkage.
No IL-2 activity prior to in vivo release
The conjugate has no interleukin-2 activity prior to release of the branched poly(ethylene glycol) polymers in vivo following administering.
Limited number of PEG arms per IL-2 moiety
The conjugate includes four, five, six, or seven branched poly(ethylene glycol) polymers covalently attached through a releasable linkage to an amino group of the interleukin-2 moiety.
Mixture with an average of six PEG polymers per IL-2 moiety
The plurality of conjugates is a mixture of conjugates with an average of six branched poly(ethylene glycol) polymers per interleukin-2 moiety.
Specified cancer indications
The cancer is selected from renal cell carcinoma, metastatic melanoma, acute myeloid leukemia, non-Hodgkin's lymphoma, cutaneous T-cell lymphoma, breast cancer and bladder cancer.
The claim coverage centers on administering an interleukin-2 (SEQ ID NO:1) conjugate containing one to seven branched PEG polymers attached to an IL-2 amino group through a releasable linkage, with branched PEG molecular weight constrained to about 20,000 to about 85,000 daltons. The dependent claims further focus on releasable carbamate linkage, no IL-2 activity prior to in vivo release, narrower PEG arm-count or mixture averages, and specified cancer indications.
Stated Advantages
Reduced undesired toxicity compared with aldesleukin (PROLEUKIN®), including reduced capillary leak syndrome and impaired neutrophil function.
Conjugates show bioactivity relative to unmodified IL-2, described by a bioactivity range of 0.1% to 100% and >105%.
Improved tumor efficacy outcomes (tumor growth delay) with rIL-2 PEG conjugates versus aldesleukin.
Pharmacokinetic improvement indicated by extended detection half-life for conjugates versus aldesleukin.
Functional activity/potency differences are reported between releasable versus unreleased conjugates and across conjugate species.
Documented Applications
Administration to treat cancer types selected from renal cell carcinoma, metastatic melanoma, acute myeloid leukemia, non-Hodgkin's lymphoma, cutaneous T-cell lymphoma, breast cancer and bladder cancer.
Tumor efficacy in lung metastatic melanoma (B16F10) model and subcutaneous melanoma model using rIL-2 PEG conjugates versus aldesleukin.
Mouse pharmacokinetics for PEG conjugates using ELISA detection, compared with aldesleukin.
In vitro cell proliferation assays using CTLL-2 cells to compare releasable versus unreleased conjugates and report EC50/potency differences across conjugate species.
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