Extracellular Matrix Binding Compounds for the Localized Loading of Therapeutic or Diagnostic Agents

The fusion protein or conjugated protein according to any one of the preceding claims, wherein the fusion protein or conjugated protein comprises or consists of an IgG Fc-homodimer, wherein each of the monomer of the IgG Fc-homodimer is C-terminally linked to at least one polypeptide (P1) (i) consisting of at least one FSD1 domain comprising or consisting of a polypeptide having at least 70% such as at least 80%, for instance at least 90%, such as at least 95%, for instance at least 99% sequence identity to SEQ ID NO:

• 1,
• and wherein each of the monomer of the IgG Fc-homodimer is linked N-terminally to a therapeutic or diagnostic moiety identical or different.

The fusion protein or conjugated protein according to any one of the preceding claims, wherein the fusion protein or conjugated protein comprises or consists of an IgG Fc-heterodimer, wherein each of the monomer of the IgG Fc-heterodimer is linked C-terminally to at least one polypeptide (P1) (i) consisting of at least one FSD1 domain comprising or consisting of a polypeptide having at least 70% such as at least 80%, for instance at least 90%, such as at least 95%, for instance at least 99% sequence identity to SEQ ID NO:

• 1,
• and wherein each of the monomer of the IgG Fc-heterodimer is N-terminally linked to a therapeutic or diagnostic moiety identical or different.

The fusion protein or conjugated protein according to any one of the preceding claims, wherein at least one of the monomers of the IgG Fc-heterodimer is linked N-terminally to at least one of the one or more polypeptide(s) (P1) (i), for example at least two polypeptides (P1), such as at least three polypeptides (P1), for example at least four polypeptides, such as at least five polypeptides (P1) consisting of at least one FSD1 domain comprising or consisting of a polypeptide having at least 70% such as at least 80%, for instance at least 90%, such as at least 95%, for instance at least 99% sequence identity to SEQ ID NO:

• 1.

The fusion protein or conjugated protein according to any one of the preceding claims wherein the therapeutic or diagnostic agent of any of the preceding claims, or the at least one the therapeutic or diagnostic moiety identical or different of any one of the preceding claims is one or more cytokine receptor.

The fusion protein or conjugated protein according to any one of the preceding claims wherein the therapeutic or diagnostic agent of any of the preceding claims, or the at least one the therapeutic or diagnostic moiety identical or different of any of the preceding claims comprises or consists of the ligand binding domain of TNFR2.

The fusion protein or conjugated protein according to any one of the preceding claims, wherein the one or more polypeptide(s) (P1) consisting of at least one FSD1 domain comprising or consisting of a polypeptide having at least 70% sequence identity, such as at least 80% identity, for instance at least 90% identity, such as at least 95% identity, for instance 99% identity to SEQ. ID. NO:

• 26.

The fusion protein or conjugated protein according to any one of the preceding claims, wherein the one or more polypeptide(s) (P1) (i) consist of a sequence having at least 70% similarity, such as at least 80% similarity, for instance at least 90% similarity, such as at least 95% similarity, for instance at least 99% similarity with a sequence selected from the group consisting of:

• SEQ ID NO:
• 23 and SEQ ID NO:
• 24.

The fusion protein or conjugated protein according to any one of the preceding claims, wherein the one or more polypeptide(s) (P1) are encoded by a sequence comprising or consisting of a sequence having at least 70% similarity, such as at least 80% similarity, for instance at least 90% similarity, such as at least 95% similarity, for instance at least 99% similarity with SEQ ID NO:

• 25.

The fusion protein or conjugated protein according to any one of the preceding claims, wherein the one or more polypeptide(s) (P1) (i), do not neutralize the activity of myostatin (SEQ ID NO:

• 45), GDF11 (SEQ ID NO:
• 46) and/or activin A (SEQ ID NO:
• 44).

The fusion protein or conjugated protein according to any one of the preceding claims, wherein said fusion protein or conjugated protein do not neutralize the activity of myostatin (SEQ ID NO:

• 45), GDF11 (SEQ ID NO:
• 46) and/or activin A (SEQ ID NO:
• 44) in a concentration range up to at least 5 nM, such as at least 7.5 nM, for example at least 8 nM, such as at least 8.2 nM, for example at least 8.4 nM, such as at least 8.6 nM, for example at least 8.8 nM, such as at least 9 nM, for example at least 10 nM, such as at least 12 nM, for example at least 15 nM, such as at least 20 nM, for example at least 30 nM, such as at least 40 nM, for example at least 50 nM, such as at least 60 nM, for example at least 61 nM, such as at least 62 nM, for example at least 63 nM, such as at least 64 nM, for example at least 64 nM, such as at least 65 nM, for example at least 70 nM, such as at least 80 nM, for example at least 100 nM, such as at least 150 nM, for example at least 200 nM, such as at least 500 nM, for example at least 600 nM, such as at least 700 nM, for example at least 750 nM, such as at least 800 nM, for example at least 820 nM, such as at least 821 nM, for example at least 822 nM, such as at least 830 nM, for example at least 1 μM, such as at least 2 μM, wherein the neutralization, or absence thereof, is determined by reporter gene bioassay of phosphorylated Smad2/3 signaling

The fusion protein or conjugated protein according to any one of the preceding claims where said fusion protein or conjugated protein do not neutralize the activity of myostatin, GDF11 and/or activin A in a concentration range up to at least 5 nM, such as at least 7.5 nM, for example at least 8 nM, such as at least 8.2 nM, for example at least 8.4 nM, such as at least 8.6 nM, for example at least 8.8 nM, such as at least 9 nM, for example at least 10 nM, such as at least 12 nM, for example at least 15 nM, such as at least 20 nM, for example at least 30 nM, such as at least 40 nM, for example at least 50 nM, such as at least 80 nM, for example at least 100 nM, such as at least 150 nM, for example at least 200 nM.

The fusion protein or conjugated protein according to any one of the preceding claims, wherein the one or more polypeptide(s) (P1) (i) do not neutralize the activity of myostatin (SEQ ID NO:

• 45), GDF11 (SEQ ID NO:
• 46) and/or activin A (SEQ ID NO:
• 44) in a concentration range up to at least 5 nM, such as at least 7.5 nM, for example at least 8 nM, such as at least 8.2 nM, for example at least 8.4 nM, such as at least 8.6 nM, for example at least 8.8 nM, such as at least 9 nM, for example at least 10 nM, such as at least 12 nM, for example at least 15 nM, such as at least 20 nM, for example at least 30 nM, such as at least 40 nM, for example at least 50 nM, such as at least 60 nM, for example at least 61 nM, such as at least 62 nM, for example at least 63 nM, such as at least 64 nM, for example at least 64 nM, such as at least 65 nM, for example at least 70 nM, such as at least 80 nM, for example at least 100 nM, such as at least 150 nM, for example at least 200 nM, such as at least 500 nM, for example at least 600 nM, such as at least 700 nM, for example at least 750 nM, such as at least 800 nM, for example at least 820 nM, such as at least 821 nM, for example at least 822 nM, such as at least 830 nM, for example at least 1 μM, such as at least 2 μM, wherein the neutralization, or absence thereof, is determined by reporter gene bioassay of phosphorylated Smad2/3 signaling

The fusion protein or conjugated protein according to any one of the preceding claims where in the one or more polypeptide(s) (P1) (i) do not neutralize the activity of myostatin, GDF11 and/or activin A in a concentration range up to at least 5 nM, such as at least 7.5 nM, for example at least 8 nM, such as at least 8.2 nM, for example at least 8.4 nM, such as at least 8.6 nM, for example at least 8.8 nM, such as at least 9 nM, for example at least 10 nM, such as at least 12 nM, for example at least 15 nM, such as at least 20 nM, for example at least 30 nM, such as at least 40 nM, for example at least 50 nM, such as at least 80 nM, for example at least 100 nM, such as at least 150 nM, for example at least 200 nM.

The fusion protein or conjugated protein according to any one of the preceding claims, wherein the one or more polypeptide(s) (P1) (i) bind heparan sulfate.

The fusion protein or conjugated protein according to any one of the preceding claims, wherein the one or more polypeptide(s) (P1) (i) comprising at least one FSD1 domain are encoded by a sequence comprising or consisting of a sequence having at least 70% identity, such as at least 80% identity, for instance at least 90% identity, such as at least 95% identity, for example at least 99% identity with SEQ. ID. NO:

• 33.

The fusion protein or conjugated protein according to any one of the preceding claims, wherein the one or more polypeptide(s) (P1) (i) comprising at least one FSD1 domain comprise or consist of a sequence having 100% identity with SEQ. ID. NO:

• 1.

The fusion protein or conjugated protein according to any one of the preceding claims, further comprising a linker between the one or more polypeptide(s) (P1) (i) consisting of at least one FSD1 domain and the therapeutic or diagnostic agent (ii).

The fusion protein or conjugated protein according to any one of the preceding claims, wherein the therapeutic or diagnostic agent (ii), or the therapeutic or diagnostic moiety identical or different of any one of the preceding claims, is selected from the group consisting of:

• a peptide, a protein, such as an antibody or fragment thereof, a nanobody, a glycoprotein, a streptavidin or an interleukin, a nucleic acid, and a small molecule.

The fusion protein or conjugated protein according to any one of the preceding claims, wherein the fusion protein or conjugated protein further comprises a detectable moiety.

The fusion protein or conjugated protein according to any one of the preceding claims, wherein the therapeutic or diagnostic agent (ii), or the therapeutic or diagnostic moiety identical or different of any one of the preceding claims binds a target selected from the group consisting of:

• cluster of differentiation (CD) proteins, cytokines, such as interleukins, growth factors, such as colony-stimulating factors, immune checkpoint proteins, angiogenic factors, hemostatic factors, chemotaxic factors, neurotrophic factors, inflammatory proteins, tumor antigens, bacterial proteins, and viral proteins.

The fusion protein or conjugated protein according to any one of the preceding claims wherein the fusion protein or conjugated protein is ubiquitinylated.

The fusion protein or conjugated protein according to any one of the preceding claims, wherein the therapeutic or diagnostic agent (ii), or the therapeutic or diagnostic moiety identical or different of any one of the preceding claims, bind to a target and wherein the fusion protein or conjugated protein according to any one of the preceding claims is able to bind heparan sulfate and said target at the same time.

The fusion protein or conjugated protein according to any one of the preceding claims, wherein the fusion protein or conjugated protein has a longer local half-life at the site of administration than the therapeutic or diagnostic agent (ii), or the therapeutic or diagnostic moiety identical or different of any one of the preceding, alone.

The fusion protein or conjugated protein according to any one of the preceding claims, wherein the fusion protein or conjugated protein increases the binding of the therapeutic or diagnostic agent (ii), or of the therapeutic or diagnostic moiety identical or different of any one of the preceding claims, to the extracellular matrix compartment compared to the therapeutic or diagnostic agent (ii), or the therapeutic or diagnostic moiety identical or different of any one of the preceding claims alone.

The fusion protein or conjugated protein according to any one of the preceding claims, wherein the fusion protein or conjugated protein increases the binding of the therapeutic or diagnostic agent (ii), or of the therapeutic or diagnostic moiety identical or different of any one of the preceding claims, to a predefined organ or a tumor compared to the therapeutic or diagnostic agent (ii), or the therapeutic or diagnostic moiety identical or different of any one of the preceding claims alone.

The fusion protein or conjugated protein according to any one of the preceding claims, wherein the EC50 of the therapeutic or diagnostic agent (ii) or the therapeutic or diagnostic moiety identical or different, to a predefined organ or a tumor, compared to the therapeutic or diagnostic agent (ii) or the therapeutic or diagnostic moiety identical or different alone is decreased by at least a factor 2, such at least a factor 5, for example at least a factor 10, such as at least a factor 20, for example at least a factor 50, such as at least a factor 80, for example at least a factor 100, such as at least a factor 250, for example at least a factor 500, such at least a factor 1000, for example at least a factor 5000, such as at least a factor 10000, for example at least a factor 100000.

The fusion protein or conjugated protein according to any of the preceding claims, wherein the fusion protein or the conjugated protein increases the intracellular uptake of the therapeutic or diagnostic agent (ii), or of the therapeutic or diagnostic moiety identical or different of any one of the preceding claims compared to the therapeutic or diagnostic agent (ii) of any one of the preceding claims, or the therapeutic or diagnostic moiety identical or different of any one of the preceding claims alone.

The fusion protein or conjugated protein according to any one of the preceding claims, wherein the therapeutic or diagnostic agent (ii) of any of the preceding claims, or the therapeutic or diagnostic moiety identical or different of any one of the preceding claims is linked to at least one polypeptide (P1) (i), for example at least two polypeptides (P1) (i), such as at least three polypeptides (P1) (i), for example at least four polypeptides, such as at least five polypeptides (P1) (i) consisting of at least one FSD1 domain comprising or consisting of a polypeptide having at least 70% such as at least 80%, for instance at least 90%, such as at least 95%, for instance at least 99% sequence identity to SEQ ID NO:

• 1.

The fusion protein or conjugated protein according to any one of the preceding claims, wherein at least one of the at least one polypeptide (P1) (i) comprises or consists of a multimer of FSD1 domains, such as at least two FSD1 domains, for example at least three FSD1 domains, for example at least four FSD1 domains, such as at least five FSD1 domains consisting of a polypeptide having at least 70% such as at least 80%, for instance at least 90%, such as at least 95%, for instance at least 99% sequence identity to SEQ ID NO:

• 1.

The fusion protein or conjugated protein according to any one of the preceding claims, wherein the fusion protein or conjugated protein does not comprise any other of the follistatin domains FSD2, FSD3, and/or a N-terminal follistatin domain, preferably wherein the fusion protein or conjugated protein does not comprise any of the human FSD2 of SEQ ID NO:

• 41, human FSD3 of SEQ ID NO:
• 42 and human follistatin N-terminal domain of SEQ ID NO:
• 43.

The fusion protein or conjugated protein according to any one of the preceding claims, wherein the one or more polypeptides (P1) (i) do not comprise any other of the follistatin domains FSD2, FSD3, and/or a N-terminal follistatin domain, preferably wherein the fusion protein or conjugated protein does not comprise any of the human FSD2 of SEQ ID NO:

• 41, human FSD3 of SEQ ID NO:
• 42 and human follistatin N-terminal domain of SEQ ID NO:
• 43.

The fusion protein or conjugated protein according to any one of the preceding claims, wherein the fusion protein or conjugated protein further comprises a PROTAC linker, preferably wherein the PROTAC linker is selected from the group consisting of:

• flexible aliphatic linkers, flexible PEGylated linkers, flexible heterochains, rigid linkers, triazole-based linkers, Bio-orthogonal clickable linkers and photo-switchable linkers.

One or more polynucleotides which upon expression encode the fusion protein or conjugated protein according to any one of the preceding claims.

One or more constructs or vectors encoding the fusion protein according to any of the preceding claims.

The method according to any one of the preceding claims, further comprising a step of administering the fusion protein or conjugated protein to a subject suffering from an ophthalmologic condition, and wherein the fusion protein or conjugated protein, or the composition is administered intravitreally, subretinally or suprachoroidally.

A method of degrading an intracellular protein, said method comprising the step of coupling the fusion protein or conjugated protein according to any one of the preceding claims to a ligand of E3 ubiquitin ligase, wherein the therapeutic or diagnostic agent (ii) of said fusion protein or conjugated protein binds to said intracellular protein.

One or more constructs or vectors encoding the one or more polypeptide(s) (P1) (i) of the fusion protein or conjugated protein according to any one of the preceding claims.