Macropinocytosis selective monobody-drug conjugates
Inventors
RAMIREZ, Craig • HAUSER, Andrew • BEALS, Nathan • Bar-Sagi, Dafna • Koide, Akiko • Koide, Shohei
Assignees
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Publication Number
US-12642859-B2
Publication Date
2026-06-02
Expiration Date
Abstract
The present disclosure is directed to pharmaceutical and diagnostic compositions comprising macropinocytosis selective non-binding protein-drug conjugates. These non-binding protein-drug conjugates comprise a non-binding fibronectin type III (FN3) domain coupled to a pharmaceutically active moiety or a diagnostic moiety. The disclosure is also directed to methods of treatment and diagnosis that involve administering the pharmaceutical compositions described herein to a subject in need.
Core Innovation
The disclosed invention provides macropinocytosis-selective non-binding protein-drug conjugates and related pharmaceutical and diagnostic compositions. Each conjugate includes a non-binding human fibronectin type III (FN3) domain paired with an amino acid linker and a second portion selected as either a pharmaceutically active moiety or a diagnostic moiety.
The non-binding FN3 domain is defined using amino acid sequence variants of SEQ ID NO: 2 or SEQ ID NO: 3, with SEQ ID NO: 2 characterized by not comprising SEQ ID NO: 1. The disclosure also includes non-binding FN3 variants, including RGD-eliminating variants, and describes optional substitutions within FN3 regions and optional tandem FN3 configurations.
The conjugate second portion is selected to support therapeutic or diagnostic use. Therapeutic payloads include MMAE, SN-38, PROTACs, and immunomodulatory agents, while diagnostic moieties include fluorophores and MRI chelates such as DOTA-Gd3+ and radiolabels. The disclosure ties the conjugate design to enhanced macropinocytosis targeting, including oncogenic Ras-mutant cancers, and to diagnostic imaging use with FN-Cy5.5 and FN-DOTA-Gd3+.
The problem being solved is selective delivery that depends on macropinocytosis, while reducing non-specific toxicity relative to free MMAE and enabling tumor-selective cellular uptake and biodistribution. The disclosure further addresses selective tumor/lymph node biodistribution in relevant cancer models, including PDAC models.
Claims Coverage
The independent claim provides a pharmaceutical composition comprising a non-binding protein-drug conjugate and a pharmaceutically acceptable carrier, with the conjugate defined by a non-binding human FN3 domain using SEQ ID NO: 2 or SEQ ID NO: 3, an amino acid linker, and a second portion selected from a pharmaceutically active moiety or a diagnostic moiety. The claims further narrow FN3 variant specificity, linker properties, and the second portion between therapeutic and diagnostic categories.
Non-binding human FN3 domain with defined sequences
A non-binding protein-drug conjugate includes a non-binding human fibronectin type III (FN3) domain, wherein the non-binding FN3 domain comprises the amino acid sequence of SEQ ID NO: 2 or SEQ ID NO: 3, and wherein the amino acid sequence of SEQ ID NO: 2 does not comprise SEQ ID NO: 1.
Amino acid linker within or coupled to the FN3 portion
An amino acid linker within or coupled to the first portion.
Second portion as pharmaceutically active moiety or diagnostic moiety
A second portion coupled to said first portion via the amino acid linker, said second portion selected from a pharmaceutically active moiety or a diagnostic moiety.
Defined FN3 substitutions in RGD-related regions
The FN3 domain includes amino acid substitutions in specified regions of its RGD sequence, including the FG loop and the BC loop sequence.
Cysteine-based linker incorporation
The amino acid linker includes a cysteine residue substitution within the FN3 domain of the first portion or includes a cysteine residue addition at the FN3 domain C-terminus.
Cleavable linker
The amino acid linker is a cleavable linker.
Cancer therapeutic-class pharmaceutically active moiety
The pharmaceutically active moiety is a cancer therapeutic selected from a specified group of therapeutic classes, including antimetabolite, alkaloid, alkylating agent, anti-mitotic agent, antitumor antibiotic, DNA binding drug, toxin, antiproliferative drug, DNA antagonist, radionuclide, thermoablative agent, PROTAC, nucleic acid inhibitor, and immune-modulatory agent.
Diagnostic moiety as second portion
The second portion of the non-binding protein-drug conjugate is a diagnostic moiety.
Across the claims, the coverage centers on a pharmaceutical composition having a non-binding protein-drug conjugate built from a non-binding human FN3 domain defined by SEQ ID NO: 2 or SEQ ID NO: 3, an amino acid linker, and a second portion chosen as either a pharmaceutically active moiety or a diagnostic moiety, with dependent claims further narrowing FN3 substitution patterns, cysteine-based linker features, cleavable linker selection, and the therapeutic-versus-diagnostic selection of the second portion.
Stated Advantages
Reduced non-specific toxicity versus free MMAE.
Enhanced macropinocytosis-dependent cellular uptake for tumor targeting.
Ras-status-dependent cytotoxicity, including reduced IC50 in Ras-mutant cells.
Selective tumor/lymph node biodistribution, including in PDAC models.
Imaging support using FN-Cy5.5 and FN-DOTA-Gd3+.
Documented Applications
Pharmaceutical composition use for tumor targeting with macropinocytosis-selective non-binding protein-drug conjugates, including oncogenic Ras-mutant cancers.
Diagnostic imaging using conjugates such as FN-Cy5.5 and FN-DOTA-Gd3+ (MRI chelates like DOTA-Gd3+).
Use with selective tumor/lymph node biodistribution in cancer models, including PDAC models.
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