Covalently-modified steroid acid-peptides having enhanced stability and/or biological activity
Inventors
Assignees
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-12233130-B2
Publication Date
2025-02-25
Expiration Date
Abstract
Steroid acid-peptide conjugates covalently modified for improved stability and/or biological activity are described herein. Covalent modifications include the formation of multimeric compounds comprising at least two steroid acid-peptide monomers covalently bound to one another that behave as new chemical entities, as well as protecting one or more free thiol groups present in the steroid acid-peptide conjugates to improve their stability and/or biological activity.
Core Innovation
The invention relates to multimeric steroid acid-peptide conjugates formed from two steroid acid-peptide monomers covalently bound to one another via an intermolecular disulfide bond. Each monomer comprises a peptide domain that includes a nuclear localization signal (NLS), and the steroid acid, in particular a bile acid or bile acid analog, mediates endosomal escape of the multimeric compound.
The invention further addresses stability and intracellular activity by protecting free thiol groups with cleavable or non-cleavable protecting groups. This protection enables enhanced stability and/or intracellular activation, including intracellular cleavage of a protecting group. The multimeric architecture is presented in the context of thiol-containing monomers forming dimer impurities, and the multimeric form exhibits improved properties relative to the monomer form.
A functional rationale is provided in which steroid acids drive intracellular endocytosis and endosomal escape through acid sphingomyelinase-mediated ceramide accumulation on the inner leaflet of endosomes. The resulting endosomal escape is associated with cytotoxic/cytostatic effects and immunogenic cell death, including apoptosis and reactive oxygen species and involvement of immunogenic markers. The peptides are described as optionally including protein transduction domains and subcellular targeting domains to support intracellular trafficking and activity.
Claims Coverage
The partial content provides one independent claim. The inventive features focus on a specific multimeric bile acid-peptide conjugate architecture, an intermolecular disulfide bond, an NLS-containing peptide, and a bile acid-mediated endosomal escape function, with dependent refinements specifying bile acid and NLS selections and additional constructs and use contexts.
Disulfide-linked bile acid-peptide NLS multimer with endosomal escape
A multimeric compound comprising two monomers covalently bound to one another via an intermolecular disulfide bond, each monomer comprising a bile acid-peptide conjugate, wherein the peptide comprised in the monomers comprises a nuclear localization signal (NLS), and wherein the bile acid comprised in the monomers mediates endosomal escape of the multimeric compound.
Sequence-defined NLS and selected bile acids
The multimeric compound further comprising wherein the bile acid is CA or CDCA and the NLS is one of hnRNPA1 M9 NLS, SV40 NLS, or hnRNP D NLS.
Bile acid analogs for ceramide accumulation
The multimeric compound further comprising wherein the bile acid is a bile acid analog of CA, CDCA, DCA, LCA, GDCA, GCA, TCA, GCDCA, TDCA, GLCA, TLCA, THDCA, TCDCA, UCA1, TUDCA, UDCA, or GUDCA that triggers ceramide accumulation on the inner leaflet of endosomes.
Conjugation to a biocompatible carrier molecule
The multimeric compound further comprising wherein the multimeric compound is conjugated to a biocompatible carrier molecule.
Antigen admixed or covalently conjugated to improve immunogenicity
The multimeric compound further comprising an antigen admixed therewith or covalently conjugated thereto, where this improves the antigen’s immunogenicity.
Pharmaceutical composition at cytotoxic or cytostatic concentration
The multimeric compound further comprising wherein the multimeric compound is comprised in a pharmaceutical composition at a cytotoxic or cytostatic concentration, free or bound to a biocompatible carrier molecule.
The claim set centers on a disulfide-linked dimeric bile acid-peptide conjugate where each monomer contains an NLS and the bile acid mediates endosomal escape. Dependent features specify particular bile acids and sequence-defined NLS variants, alternative bile acid analogs linked to ceramide accumulation, attachment to a biocompatible carrier molecule, optional inclusion of an antigen to improve immunogenicity, and use in pharmaceutical compositions at cytotoxic or cytostatic concentrations.
Stated Advantages
Improves the antigen’s immunogenicity.
Enhanced stability of the multimeric compound relative to thiol-containing monomer forms, described as markedly higher stability.
Enhanced cytotoxicity, described as enhanced cytotoxicity with lower EC50 versus monomers.
Improves antigen cross-presentation.
Documented Applications
Cytotoxic/cytostatic therapy using dimeric steroid acid-peptide conjugates in pharmaceutical compositions at cytotoxic or cytostatic concentrations.
Immunogenic cell death with antigen presentation, including improved antigen cross-presentation.
Antigen presentation and vaccine use where an antigen is admixed or covalently conjugated to improve immunogenicity.
In vivo tumor model activity against EL4 lymphoma, B16F10 melanoma, and E0771 breast cancer.
Combination with immune-checkpoint inhibitors including anti-PD-1 and anti-CTLA4, and described interactions with immune-checkpoint targets such as anti-CD47 and anti-LAG3.
Interested in licensing this patent?