Thermoresponsive bioconjugates and their controlled delivery of cargo

• US 10,639,371 B2
• Filed: 07/29/2016
• Issued: 05/05/2020
• Est. Priority Date: 07/29/2015
• Status: Active Grant

• Current Assignee: University of Delaware
• Original Assignee: University of Delaware
• Inventors: Kiick, Kristi, Luo, Tianzhi
• Primary Examiner(s): Gudibande, Satyanarayana R
• Application Number:
  US15/747,552
• Publication Number:
  US 20180236074A1
• Time in Patent Office: 1,376 Days
• Field of Search: None
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 38/39 Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin, cold insoluble globulin [CIG]
• CPC Class Codes: A61K 41/0028 Disruption, e.g. by heat or ultrasounds, sonophysical or sonochemical activation, e.g. thermosensitive or heat-sensitive liposomes, disruption of calculi with a medicinal preparation and ultrasounds
• CPC Class Codes: A61K 47/62 the modifying agent being a protein, peptide or polyamino acid
• CPC Class Codes: A61K 47/6907 the form being a microemulsion, nanoemulsion or micelle
• CPC Class Codes: A61K 47/6931 the material constituting the nanoparticle being a polymer
• CPC Class Codes: A61K 47/6935 the polymer being obtained otherwise than by reactions involving carbon to carbon unsaturated bonds, e.g. polyesters, polyamides or polyglycerol
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 14/78 Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]

Abstract

The present invention provides a bioconjugate. The bioconjugate comprises a first molecule comprising an elastin-like peptide (ELP), a second molecule capable of self-associating into an oligomer, and a linker connecting the first molecule with the second molecule. The bioconjugate is self-assembled into particles, which are disassembled when the temperature is increased. Also provided is a method for delivering a bioconjugate to a target matrix, comprising (a) introducing a bioconjugate self-assembled into particles to a target matrix, wherein the bioconjugate comprises a first molecule comprising an elastin-like peptide (ELP), a second molecule capable of self-associating into an oligomer, and a linker connecting the first molecule with the second molecule, and (b) increasing the temperature of the target matrix to disassemble the particles at the target matrix.

Claims

1. A method for delivering a thermoresponsive bioconjugate to a target matrix, comprising

(a) introducing a thermoresponsive bioconjugate self-assembled into particles to a target matrix, wherein the bioconjugate comprises a first molecule comprising an elastin-like peptide (ELP) having no more than 125 amino acids, a second molecule capable of forming an oligomer, and a linker connecting the first molecule with the second molecule, and wherein the ELP has 3-10 repeats of VPGXG (SEQ ID NO:

22), XPGVG (SEQ ID NO;

     28) or XPAVG (SEQ ID NO;

     27), wherein X is an amino acid, and

(b) increasing or decreasing the temperature of the target matrix, whereby the particles are disassembled at the target matrix.

2. The method of claim 1, wherein the ELP is VPGXGVPGXGVPGXG (SEQ ID NO:

23), VPGXGVPGXGVPGXGVPGXG (SEQ ID NO;

     24), VPGXGVPGXGVPGXGVPGXGVPGXG (SEQ ID NO;

     25) or VPGXGVPGXGVPGXGVPGXGVPGXGVPGXG (SEQ ID NO;

     26), wherein X is an amino acid.

3. The method of claim 1, wherein the ELP is VPGFGVPGFGVPGFGVPGFGVPGFGVPGFG (SEQ ID NO:

21).

4. The method of claim 1, wherein the second molecule comprises a peptide.

5. The method of claim 4, wherein the peptide is a collagen-like peptide (CLP).

6. The method of claim 5, wherein the CLP has 5-15 repeats of GPO or GPP, wherein O is hydroxyproline.

7. The method of claim 5, wherein the CLP is selected from the group consisting of GPOGPOGPOGFOGERGPOGPOGPO (SEQ ID NO:

29), GPOGPOGPOGPOGFOGERGPOGPOGPOGPO (SEQ ID NO;

     8), GPOGPOGPOGPOGPOGFOGERGPOGPOGPOGPOGPO (SEQ ID NO;

     30), GPPGPPGPPGPPGFOGERGPPGPPGPPGPP (SEQ ID NO;

     31), GPPGPPGPPGPPGPPGFOGERGPPGPPGPPGPPGPP (SEQ ID NO;

     12), GPOGPOGPOGEKGERGPOGPOGPO (SEQ ID NO;

     6), GPOGPOGPOGPOGEKGERGPOGPOGPOGPO (SEQ ID NO;

     7), GPOGPOGPOGPOGPOGEKGERGPOGPOGPOGPOGPO (SEQ ID NO;

     9), GPPGPPGPPGEKGERGPPGPPGPP (SEQ ID NO;

     10) and GPPGPPGPPGPPGEKGERGPPGPPGPPGPP (SEQ ID NO;

     11), wherein 0 is hydroxyproline.

8. The method of claim 5, wherein the CLP is GPOGPOGPOGPOGFOGERGPOGPOGPOGPO (SEQ ID NO:

8), wherein O is hydroxyproline.

9. The method of claim 4, wherein the peptide is a coiled-coil peptide (CCP).

10. The method of claim 9, wherein the CCP is selected from the group consisting of RMKQIEDKLEEILSKLYHIENELARIKKLLGER (SEQ ID NO:

15), LKALEEKLKALEEKLKALEEK (SEQ ID NO;

     14), KLKELKSKLKELLKLELQAIKQYKELKAEKLEL (SEQ ID NO;

     13), TQEDLLKKIMKLLKKQIKLLKKQIKMLKRLEKQ (SEQ ID NO;

     17), SDLGPQMLRELQETNAALQDVRDWLRQQVREITFLKNTVMECDACG (SEQ ID NO;

     16), GEQTKALVTQLTLFNQILVELRDDIRDQVKEMSLIRNTIMECQVCG (SEQ ID NO;

     4), GDFNRQFLGQMTQLNQLLGEVKDLLRQQVKETSFLRNTIAECQACG (SEQ ID NO;

     3), ASTDTLQAETDQLEDEKYALQTEIANLLKEKEKLGAP (SEQ ID NO;

     2) and ASIARLEEKVKTLKAQNYELASTANMLREQVAQLGAP (SEQ ID NO;

     1).

11. The method of claim 1, wherein the bioconjugate has formula (I):

19. The method of claim 1, wherein the temperature of the target matrix is increased.

20. The method of claim 1, wherein the temperature of the target matrix is decreased.

12. A thermoresponsive bioconjugate comprising a first molecule comprising an elastin-like peptide (ELP) having no more than 125 amino acids, a second molecule capable of forming an oligomer, and a linker connecting the first molecule with the second molecule, wherein the ELP has 3-10 repeats of VPGXG (SEQ ID NO:

22), XPGVG (SEQ ID NO;

     28) or XPAVG (SEQ ID NO;

     27), wherein X is an amino acid, wherein the bioconjugate is self-assembled into particles, and wherein the particles are disassembled when the temperature is increased or decreased.

13. The bioconjugate of claim 12, wherein the ELP is VPGXGVPGXGVPGXG (SEQ ID NO:

23), VPGXGVPGXGVPGXGVPGXG (SEQ ID NO;

     24), VPGXGVPGXGVPGXGVPGXGVPGXG (SEQ ID NO;

     25) or VPGXGVPGXGVPGXGVPGXGVPGXGVPGXG (SEQ ID NO;

     26), wherein X is an amino acid.

14. The bioconjugate of claim 12, wherein the ELP is VPGFGVPGFGVPGFGVPGFGVPGFGVPGFG (SEQ ID NO:

21).

15. The bioconjugate of claim 12, wherein the second molecule comprises a peptide.

16. The bioconjugate of claim 15, wherein the peptide is a collagen-like peptide (CLP).

17. The bioconjugate of claim 16, wherein the CLP has 5-15 repeats of GPO or GPP, wherein O is hydroxyproline.

18. The bioconjugate of claim 16, wherein the CLP is selected from the group consisting of GPOGPOGPOGFOGERGPOGPOGPO (SEQ ID NO:

29), GPOGPOGPOGPOGFOGERGPOGPOGPOGPO (SEQ ID NO;

     8), GPOGPOGPOGPOGPOGFOGERGPOGPOGPOGPOGPO (SEQ ID NO;

     30), GPPGPPGPPGPPGFOGERGPPGPPGPPGPP (SEQ ID NO;

     31), GPPGPPGPPGPPGPPGFOGERGPPGPPGPPGPPGPP (SEQ ID NO;

     12), GPOGPOGPOGEKGERGPOGPOGPO (SEQ ID NO;

     6), GPOGPOGPOGPOGEKGERGPOGPOGPOGPO (SEQ ID NO;

     7), GPOGPOGPOGPOGPOGEKGERGPOGPOGPOGPOGPO (SEQ ID NO;

     9), GPPGPPGPPGEKGERGPPGPPGPP (SEQ ID NO;

     10) and GPPGPPGPPGPPGEKGERGPPGPPGPPGPP (SEQ ID NO;

     11).