Enzyme-responsive peptide nanofiber compositions and uses thereof

• US 10,449,259 B2
• Filed: 09/30/2016
• Issued: 10/22/2019
• Est. Priority Date: 10/02/2015
• Status: Active Grant

• Current Assignee: Cornell University
• Original Assignee: Cornell University
• Inventors: Law, Shek Hang Benedict, Tung, Ching-Hsuan, Bellat, Vanessa
• Primary Examiner(s): Cordero Garcia, Marcela M
• Application Number:
  US15/764,188
• Publication Number:
  US 20180296697A1
• Time in Patent Office: 1,117 Days
• Field of Search: None
• CPC Class Codes: A61K 31/167 having the nitrogen of a carboxamide group directly attached to the aromatic ring, e.g. lidocaine, paracetamol
• CPC Class Codes: A61K 31/5365 ortho- or peri-condensed with heterocyclic ring systems
• CPC Class Codes: A61K 31/704 attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin digitoxin A61K31/7048
• CPC Class Codes: A61K 38/063 Glutathione
• CPC Class Codes: A61K 38/2013 IL-2
• CPC Class Codes: A61K 38/2073 IL-11
• CPC Class Codes: A61K 38/212 IFN-alpha
• CPC Class Codes: A61K 38/50 acting on carbon-nitrogen bonds, other than peptide bonds (3.5), e.g. asparaginase
• CPC Class Codes: A61K 47/42 Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein oligopeptides having up to five amino acids A61K47/183; polyamino acids A61K47/34
• CPC Class Codes: A61K 47/60 the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
• CPC Class Codes: A61K 47/65 Peptidic linkers, binders or spacers, e.g. peptidic enzyme-labile linkers
• CPC Class Codes: A61K 47/6953 the form being a fibre, a textile, a slab or a sheet
• CPC Class Codes: A61K 9/0024 Solid, semi-solid or solidifying implants, which are implanted or injected in body tissue compositions for intravenous administration, normal injectable solutions or dispersions for, e.g. subcutaneous administration A61K9/0019; brain implants A61K9/0085; (coated) prostheses, catheters or stents A61L
• CPC Class Codes: A61K 9/70 Web, sheet or filament bases ; Films; Fibres of the matrix type containing drug hollow drug-filled fibres A61K9/0092
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C12Y 305/01001 Asparaginase (3.5.1.1)

Abstract

The disclosure provides compositions comprising peptide-based nanofiber precursors and methods of using the same to inhibit cancerous cell growth and/or to deliver therapeutic or diagnostic agents to cells, e.g., cancerous cells. The compositions of the present technology include peptide-based nanofiber precursors as well as carrier complexes comprising a therapeutic or diagnostic agent, and a peptide-based nanofiber precursor. Also provided herein are methods for delivering a therapeutic or diagnostic agent to a cell comprising contacting the cell with a carrier complex including a therapeutic or diagnostic agent, and a peptide-based nanofiber precursor.

Claims

1. A peptide-based nanofiber comprising a plurality of peptides, wherein each peptide comprises a methoxypolyethylene glycol (mPEG) polymer, GSH, and a self-assembling domain comprising the sequence KLDLKLDLKLDLK (SEQ ID NO:

1), wherein the self-assembling domain is interspersed between GSH and the mPEG polymer, wherein each amino acid of SEQ ID NO;

1 is an L-amino acid.

2. The peptide nanofiber of claim 1, wherein the mPEG polymer is located at the N-terminus of each peptide.

3. The peptide nanofiber of claim 1, wherein the mPEG polymer has a molecular weight from 1 kDa to 5 kDa.

4. A peptide-based nanofiber comprising a mixture of a first plurality of peptides and a second plurality of peptides,

wherein each peptide of the first plurality of peptides comprises a first methoxypolyethylene glycol (mPEG) polymer, a first fluorophore, and a first self-assembling domain comprising the sequence KLDLKLDLKLDLK (SEQ ID NO:

1), wherein the first self-assembling domain is interspersed between the first fluorophore and the first mPEG polymer, wherein each amino acid of SEQ ID NO;

1 is an L-amino acid; and

wherein each peptide of the second plurality of peptides comprises a second methoxypolyethylene glycol (mPEG) polymer, GSH and a second self-assembling domain comprising the sequence KLDLKLDLKLDLK (SEQ ID NO;

     1).

5. The peptide-based nanofiber of claim 4, wherein the molar ratio of the first plurality of peptides to the second plurality of peptides is 4:

1, 5;

1, 6;

1, 7;

1, 8;

1, 9;

1, 10;

1, 11;

1, 12;

1, 13;

1, 14;

1, 15;

1, 16;

1, 17;

1, 18;

1, 19;

1, 20;

1, 25;

1, 30;

1, 40;

1, or 50;

1.

6. A peptide-based nanofiber comprising a mixture of a first plurality of peptides, a second plurality of peptides, and a third plurality of peptides,

wherein each peptide of the first plurality of peptides comprises a first methoxypolyethylene glycol (mPEG) polymer, a first fluorophore, and a first self-assembling domain comprising the sequence KLDLKLDLKLDLK (SEQ ID NO:

1), wherein the first self-assembling domain is interspersed between the first fluorophore and the first mPEG polymer, wherein each amino acid of SEQ ID NO;

1 is an L-amino acid;

wherein each peptide of the second plurality of peptides comprises a second methoxypolyethylene glycol (mPEG) polymer, and a second self-assembling domain comprising the sequence KLDLKLDLKLDLC (SEQ ID NO;

     11); and

wherein each peptide of the third plurality of peptides comprises a third methoxypolyethylene glycol (mPEG) polymer, GSH and a third self-assembling domain comprising the sequence KLDLKLDLKLDLK (SEQ ID NO;

     1).

7. The peptide-based nanofiber of claim 6, wherein the molar ratio of the third plurality of peptides to the first and second plurality of peptides is 1:

49, 2;

48, 3;

47, 4;

46, 5;

45, 6;

44, 7;

43, 8;

42, 9;

41, or 10;

40.

8. The peptide-based nanofiber of claim 6, wherein the molar ratio of the first plurality of peptides to the second plurality of peptides to the third plurality of peptides is 1:

31;

8, 1;

32;

7, 1;

33;

6, 1;

34;

5, 1;

35;

4, 1;

36;

3, 1;

37;

2, 1;

38;

1, 1;

38.1;

0.9, or 1;

31.2;

0.8.

9. The peptide-based nanofiber of claim 6, wherein the peptide-based nanofiber is surface functionalized with one or more targeting ligands selected from the group consisting of antibodies, polymers, and peptide moieties.

10. The peptide-based nanofiber of claim 9, wherein the peptide moieties are selected from the group consisting of Bombesin (BBN), Somatostatin, Allatostatin 1, Follicle-stimulating hormone analog FSH-33, LyP-1 peptide, Fibroblast growth factor analogs, Hepatocarcinoma targeting peptide, Peptide GFE, Epidermal Growth Factor, cetuximab, RGD tripeptide, CendR peptide, peptide F3, α

-MSH peptide, Enterotoxin (STh), cyclic decapeptide CGLIIQKNEC (SEQ ID NO;

     6), WIFPWIQL (SEQ ID NO;

     7) peptide, CREKA (SEQ ID NO;

     8) peptide, IPLVVPL (SEQ ID NO;

     9) peptide, KTLLPTP (SEQ ID NO;

     10) peptide, and antitumor-antibody-derived peptides.

11. A carrier complex comprising a therapeutic or diagnostic agent conjugated to a peptide-based nanofiber of claim 6.

12. The carrier complex of claim 11, wherein the therapeutic agent is selected from the group consisting of 13-cis-Retinoic Acid, 2-Chlorodeoxyadenosine, 5-Azacitidine, 5-Fluorouracil, 6-Mercaptopurine, 6-Thioguanine, actinomycin-D, adriamycin, aldesleukin, alemtuzumab, alitretinoin, all-transretinoic acid, alpha interferon, altretamine, amethopterin, amifostine, anagrelide, anastrozole, arabinosylcytosine, arsenic trioxide, amsacrine, aminocamptothecin, aminoglutethimide, asparaginase, azacytidine, Bacillus calmette-guerin (BCG), bendamustine, bevacizumab, bexarotene, bicalutamide, bortezomib, bleomycin, busulfan, calcium leucovorin, citrovorum factor, capecitabine, canertinib, carboplatin, carmustine, cetuximab, chlorambucil, cisplatin, cladribine, cortisone, cyclophosphamide, cytarabine, darbepoetin alfa, dasatinib, daunomycin, decitabine, denileukin diftitox, dexamethasone, dexasone, dexrazoxane, dactinomycin, daunorubicin, decarbazine, docetaxel, doxorubicin, doxil, aldoxorubicin, doxifluridine, edrecolomab, eniluracil, epirubicin, epoetin alfa, erlotinib, everolimus, exemestane, estramustine, etoposide, filgrastim, fluoxymesterone, fulvestrant, flavopiridol, floxuridine, fludarabine, fluorouracil, flutamide, gefitinib, gemcitabine, gemtuzumab ozogamicin, goserelin, granulocyte-colony stimulating factor, granulocyte macrophage-colony stimulating factor, hexamethylmelamine, hydrocortisone hydroxyurea, ibritumomab, ibritumomab tiuxetan, interferon alpha, interleukin-2, interleukin-11, isotretinoin, ixabepilone, idarubicin, imatinib mesylate, ifosfamide, irinotecan, lapatinib, lenalidomide, letrozole, leucovorin, leuprolide, liposomal Ara-C, lomustine, mechlorethamine, megestrol, melphalan, mercaptopurine, mertansine, mesna, methotrexate, methylprednisolone, mitomycin C, mitotane, mitoxantrone, nelarabine, nilutamide, octreotide, oprelvekin, oxaliplatin, paclitaxel, pamidronate, pemetrexed, panitumumab, PEG Interferon, pegaspargase, pegfilgrastim, PEG-L-asparaginase, pentostatin, plicamycin, prednisolone, prednisone, procarbazine, raloxifene, rituximab, romiplostim, ralitrexed, sapacitabine, sargramostim, satraplatin, sorafenib, sunitinib, semustine, streptozocin, tamoxifen, tegafur, tegafur-uracil, temsirolimus, temozolamide, teniposide, thalidomide, thioguanine, thiotepa, topotecan, toremifene, tositumomab, trastuzumab, trastuzumab emtansine, tretinoin, trimitrexate, alrubicin, vincristine, vinblastine, vindestine, vinorelbine, vorinostat, and zoledronic acid.

13. The carrier complex of claim 11, wherein the diagnostic agent is a nanoparticle, a radioactive substance, a dye, a fluorescent compound, a contrast agent, a bioluminescent compound, an enzyme, or an enhancing agent.

14. A method for delivering a therapeutic or diagnostic agent to a cancerous cell, the method comprising contacting the cell with the carrier complex of claim 11.

15. A method for delivering a therapeutic or diagnostic agent to a cancerous cell in a subject in need thereof comprising administering to the subject an effective amount of the carrier complex of claim 11.

16. The method of claim 15, wherein the subject is diagnosed with breast cancer, brain cancer, ovarian cancer, or prostate cancer.

17. The method of claim 15, wherein the subject is human.

18. The method of claim 15, wherein the peptide-based nanofiber is administered orally, topically, intranasally, systemically, locally intramuscularly, intravenously, subcutaneously, intracerebroventricularly, intrathecally, intratumorally, transdermally or with iontophoresis.

19. The peptide-based nanofiber of claim 6, wherein the first fluorophore is FITC, TAMRA, or