Therapeutic compounds for red blood cell-mediated delivery of an active pharmaceutical ingredient to a target cell

A therapeutic compound for RBC-mediated delivery in a mammalian subject to a fibrotic cell expressing CD47, the therapeutic compound comprising:

• a CD47-binding protein conjugated to an API so as to form a conjugate;
• wherein:
• the CD47-binding protein is selected from the group consisting of wild type thrombospondin-1 (TSP-1) (SEQ ID NO:
• 7), wild type SIRPγ (SEQ ID NO:
• 4), vSIRPγ-1 (SEQ ID NO:
• 5), vSIRPγ-2 (SEQ ID NO:
• 6), ALX148 (SEQ ID NO:
• 962), TTI-661 (SEQ ID NO:
• 963), TTI-662 (SEQ ID NO:
• 964), a homolog of any of the foregoing, and combinations thereof, and is configured to bind the conjugate to CD47 of a red blood cell of the subject so as to enable transport of the conjugate, through the subject's circulatory system, to the fibrotic cell, so that (i) the CD47-binding protein, being configured to bind the conjugate to the CD47 of the red blood cell, binds the CD47 of the fibrotic cell, thus transferring the conjugate from the red blood cell to the fibrotic cell so as to form a conjugate-CD47 complex on the fibrotic cell, thereby blocking CD47 and inhibiting CD47 activity as an immune escape mechanism of the fibrotic cell, and (ii) the conjugate is taken up by the fibrotic cell via endocytosis of the conjugate-CD47 complex, thereby further inhibiting the immune escape mechanism of the fibrotic cell and delivering the API into the fibrotic cell;
• and
• the API is shRNA, the shRNA being a single-stranded RNA molecule of 44-71 nucleotides in length, and having, in a 5′ to 3′ direction:
• a first region of 19-29 nucleotides at the 5′ end of the single-stranded RNA molecule, the first region having a first sequence;
• a second region of 4-11 nucleotides directly adjacent to the first region, the second region having a second sequence;
• a third region of 19-29 nucleotides directly adjacent to the second region, the third region having a third sequence;
• and
• a fourth region of 2 nucleotides at the 3′ end of the single-stranded RNA molecule, directly adjacent to the third region, the fourth region having a fourth sequence,
• wherein:
• (a) the first region has the same number of nucleotides as the third region,
• (b) the third sequence is the reverse-complement of the first sequence,
• (c) the third region is complementary to contiguous nucleotides in a target mammalian mRNA sequence, the mRNA sequence being selected from the group consisting of SEQ ID NO:
• 8-21, 40-43, and 766-770, and
• (d) the single-stranded RNA molecule is configured to form a stem loop structure, the first region base pairing with the third region to form a stem, the second region forming a loop, and the fourth region forming a 3′ overhang.