US 2024/0197846 A1 · Antigen-Presenting Polypeptides with Chemical Conjugation Sites and Methods of Use Thereof

Abstract

The present disclosure provides antigen presenting polypeptide comprising a TGF-β MOD that is reversibly masked and acts as a TGF-β receptor agonist. The antigen presenting polypeptides comprising one or more chemical conjugation sites for incorporation of, for example, Type 1 Diabetes (T1D) associated epitope containing polypeptides. The antigen-presenting polypeptides and their T1D-associated epitope conjugates are useful for modulating the activity of a T-cell, and accordingly, the present disclosure provides methods of modulating activity of a T-cell in vitro and in vivo as a method of treatment of T1D.

Claims40

Dependent claims

A TMAPP-epitope conjugate comprising:

(i) a presenting sequence or a presenting complex,
(ii) optionally at least one scaffold polypeptide sequence, and
(iii) a TGF-β sequence, a masking sequence that binds to a TGF-β sequence reversibly masking it, or at least one masked TGF-β MOD;
wherein
(a) each presenting sequence comprises MHC Class II α1, α2, β1, and β2 domain polypeptide sequences;
(b) each presenting complex comprises a presenting complex first sequence and a presenting complex second sequence, wherein
the presenting complex first sequence and presenting complex second sequence comprises at least one of the α1, α2, β1, and β2 polypeptide sequences, and
the presenting complex first sequence and presenting complex second sequence together comprise the MHC Class II α1, α2, β1, and β2 domain polypeptide sequences, and
(c) each masked TGF-β MOD comprises a masking sequence and TGF-β sequence;
wherein the TMAPP-epitope conjugate optionally comprises one or more independently selected additional MODs (wt. and/or variant) or pairs of additional MODs (e.g., in tandem, both wt, both variant, or one wt. and one variant) (e.g., located at the N-terminus of a presenting sequence, presenting complex first and/or second sequences);
wherein a the TMAPP-epitope conjugate comprises a chemical conjugation site for conjugation of an epitope presenting molecule, and optionally comprises an additional chemical conjugation site for the conjugation of a payload, and wherein a T1D-associated epitope has been conjugated to the chemical conjugation site;
and
wherein the TMAPP-epitope conjugate optionally comprises one or more linker sequences that are selected independently. (see, e.g., FIGS. 1A to 1H and FIGS. 10A to 10D).
It is understood that the amino acid sequence of any component (e.g., MHC-Class II polypeptide sequences) does not include an amino acid sequence that will anchor the TMAPP in a mammalian cell (e.g., a COS cell) membrane (e.g., the TMAPP does not comprise an MHC transmembrane domain, or a portion thereof, that will anchor the TMAPP in a cell membrane. The above components may or may not be arranged in the stated order from N-terminus to C-terminus in the TMAPP.

11 more elements

The TMAPP-epitope conjugate of any preceding claim, wherein:

(i). the MHC class II α1 and α2 domain polypeptide sequences comprise human class II α1 and α2 domain polypeptide having at least 90% or at least 95% (e.g., at least 98% or 100%) sequence identity to an HLA DR alpha (DRA), DQ alpha 1 (DQA1), or DQ alpha 2 (DQA2) α1 and α2 domain polypeptide sequences;
and
(ii). the MHC class II β1 and β2 domain polypeptide sequences comprises human class β1 and β2 domain polypeptide sequences having at least 90% or at least 95% (e.g., at least 98% or 100%) sequence identity to an HLA DR beta 1 (DRB1), DR beta 3 (DRB3), DR beta 4 (DRB4), DR beta 5 (DRB5), DQ beta 1 (DQB1), or DQ beta 2 (DQB2) β1 and β2 domain polypeptide sequences.

The TMAPP-epitope conjugate of any preceding claim, wherein:

(i) the α1 and α2 domain polypeptide sequences each have at least 90% or at least 95% (e.g., at least 98% or 100%) sequence identity to all or at least about 50 (e.g., at least about 60, 70, 80, 85, or 90) contiguous aas of the HLA DR alpha (DRA) α1 or α2 domain polypeptide sequence provided in FIG. 4;
and
(ii) the β1 and β2 domain polypeptide sequences each have at least 90% or at least 95% (e.g., at least 98% or 100%) sequence identity to all or at least about 50 (e.g., at least about 60, 70, 80, 85, or 90) contiguous aas of an HLA DR beta 1 (DRB1), DR beta 3 (DRB3), DR beta 4 (DRB4), or DR beta 5 (DRB5) 01 or β2 domain polypeptide sequence provided in any one of FIG. 5, 6, 7, or 8.

The TMAPP-epitope conjugate of any preceding claim, wherein:

(i) the α1 and α2 domain polypeptide sequences each have at least 90% or at least 95% (e.g., at least 98% or 100%) sequence identity to all or at least about 50 contiguous aas of a HLA DRA*0101 or HLA DRA*0102 polypeptide sequence;
and
(ii) the β1 and β2 domain polypeptide sequences each have at least 90% or at least 95% (e.g., at least 98% or 100%) sequence identity to at least about 60 contiguous aas of an DRB1*0301, DRB1*0401, DRB1*0402, DRB1*0405, DRB1*0801, or DRB1*0901 polypeptide sequence.

The TMAPP-epitope conjugate of any preceding claim, comprising a scaffold polypeptide sequence that is a non-interspecific sequence or interspecific sequence.

The TMAPP-epitope conjugate of any preceding claim, wherein the TGF-β sequence is:

(i) a TGF-β 1 polypeptide optionally comprising a substitution of C77;
(ii) a TGF-β2 polypeptide optionally comprising a substitution of C77;
or
(iii) is a TGF-β3 polypeptide optionally comprising a substitution of C77.

1 more element

The TMAPP-epitope conjugate or duplex TMAPP-epitope conjugate of any preceding claim, wherein the TGF-β sequence is a TGF-β3 polypeptide optionally comprising a substitution of C77.

The TMAPP-epitope conjugate or duplex TMAPP-epitope conjugate of any preceding claim, wherein the chemical conjugation site or the additional chemical conjugation site is selected from the group consisting of:

a) an amino acid chemical conjugation site;
b) non-natural amino acids and/or selenocysteines;
c) a peptide sequence that acts as an enzyme modification sequence (e.g., sulfatase, transglutaminase or sortase sites);
d) carbohydrate or oligosaccharide;
and
e) IgG nucleotide binding sites.

3 more elements

The TMAPP-epitope conjugate or duplex TMAPP-epitope conjugate of any preceding claim, wherein the additional MOD (wt. or variant) or the additional pair of MODs (wt. or variant) are selected independently from the group consisting of IL-2, PD-L1, 4-1BBL polypeptide sequences and variants of any thereof. For example, the unconjugated TMAPP or unconjugated duplex TMAPP may comprise at least one IL-2 MOD (wt. or variant) and/or at least one PD-L1 (wt. or variant) polypeptide sequence(s).

The TMAPP-epitope conjugate or duplex TMAPP-epitope conjugate of any preceding claim, wherein the additional MOD (wt. or variant) or the additional pair of MODs (wt. or variant) comprise at least one IL-2 MOD (wt. or variant) polypeptide sequence, or at least one pair of IL-2 MOD (wt. or variant) polypeptide sequences in tandem.

The TMAPP-epitope conjugate or duplex TMAPP-epitope conjugate of any preceding claim wherein the additional MOD (wt. or variant) or the additional pair of MODs (wt. or variant) comprise at least one 4-1BBL MOD (wt. or variant) polypeptide sequence.

Dependencies are inferred from the claim text.

Antigen-Presenting Polypeptides with Chemical Conjugation Sites and Methods of Use Thereof

Life of this patent

Antigen-Presenting Polypeptides with Chemical Conjugation Sites and Methods of Use Thereof

Life of this patent

Abstract

Claims40