Peptides for targeting apoptotic cells and uses thereof

Patent metadata

Field	Value
Publication number	US-8993721-B2
Application number	US-201213467689-A
Country	US
Kind code	B2
Filing date	May 9, 2012
Priority date	May 14, 2008
Publication date	Mar 31, 2015
Grant date	Mar 31, 2015

Abstract

Described herein is an isolated polypeptide which is capable of specifically targeting apoptotic cells undergoing apoptosis and consists of the sequence (I): Cys-X 1 -Val-Ala-Pro-X 2 (I), wherein X 1 is an amino acid with polar uncharged side chain and X 2 is an amino acid with positive charged side chain. The isolated polypeptide of the present invention may be useful for detecting apoptotic cells, as well as detecting and imaging apoptotic cells in tumor tissue, apoptotic myocardial cells in myocardial infarction tissue, apoptotic nerve cells in stroke tissue, and arteriosclerosis site; the polypeptide is useful for targeted drug delivery thereto.

First claim

The invention claimed is:

1. A composition for detecting apoptotic cells comprising:
(i) an isolated polypeptide consisting of the sequence (I) Cys-X1-Val-Ala-Pro-X2; and
(ii) a labeling agent bound to the polypeptide of (i) selected from the group consisting of a coloring enzyme, a radioactive isotope, and a fluorescer,
wherein X1 is an amino acid with polar uncharged side chain and X2 is an amino acid with positive charged side chain, and
wherein the coloring enzyme is peroxidase or alkaline phosphatase; the radioactive isotope is selected from the group consisting of 18F, 124I, 125I, 32P, and 35P; and the fluorescer is selected from the group consisting of Fluorescein isothiocyanate (FITC), Rhodamine isothiocyanate (RITC), Green Fluorescent Protein (GFP), Enhanced Green Fluorescent Protein (EGFP), Red Fluorescent Protein (RFP), Discosoma sp. Red Fluorescent Protein (DsRed), Cyan Fluorescent Protein (CFP), Cyan Green Fluorescent Protein (CGFP), YFP (Yellow Fluorescent Protein), Cy3, Cy5, and Cy7.5.

2. A method for detecting apoptotic cells comprising the steps of:
(a) mixing the polypeptide of claim 1 with a sample;
(b) removing unbound or unspecifically bound polypeptide; and
(c) detecting the binding and the location of the polypeptide.

3. A pharmaceutical composition for treating neoplastic disease comprising:
(i) an isolated polypeptide consisting of the sequence (I) Cys-X1-Val-Ala-Pro-X2; and
(ii) an antitumor agent bound to the polypeptide of (i) as an effective ingredient;
wherein X1 is an amino acid with polar uncharged side chain and X2 is an amino acid with positive charged side chain,
wherein the neoplastic disease is selected from the group consisting of colon cancer, lung cancer, stomach cancer, esophageal cancer, pancreatic cancer, gallbladder cancer, renal cancer, bladder cancer, prostate cancer, testicular cancer, cervical cancer, endometrial cancer, choriocarcinoma, ovarian cancer, breast cancer, thyroid cancer, brain cancer, head and neck cancer, malignant melanoma, skin cancer, liver cancer, leukemia, lymphoma, multiple myeloma, chronic myelogenous leukemia, neuroblastoma, and aplastic anemia, and
wherein the antitumor agent is selected from the group consisting of paclitaxel, doxorubicin, vincristine, daunorubicin, vinblastine, actinomycin-D, docetaxel, etoposide, teniposide, bisantrene, homoharringtonine, Gleevec (STI-571), cisplatin, 5-fluorouracil, adriamycin, methotrexate, busulfan, chlorambucil, cyclophosphamide, melphalan, nitrogen mustard, and nitrosourea.

4. A pharmaceutical composition for treating stroke comprising:
(i) an isolated polypeptide consisting of the sequence (I) Cys-X1-Val-Ala-Pro-X2; and
(ii) an anti-stroke agent bound to the polypeptide of (i) as an effective ingredient;
wherein X1 is an amino acid with polar uncharged side chain and X2 is an amino acid with positive charged side chain, and
wherein the anti-stroke agent is selected from the group consisting of streptokinase, urokinase, and alteplase.

5. A pharmaceutical composition for treating myocardial infarction comprising:
(i) an isolated polypeptide consisting of the sequence (I) Cys-X1-Val-Ala-Pro-X2; and
(ii) an anti-myocardial infarction agent bound to the polypeptide of (i) as an effective ingredient;
wherein X1 is an amino acid with polar uncharged side chain and X2 is an amino acid with positive charged side chain, and
wherein the anti-myocardial infarction agent is selected from the group consisting of streptokinase, urokinase, alteplase, angiotensin II inhibitor, aldosterone receptor inhibitor, erythropoietin, and NMDA (N-methyl-D-aspartate) receptor inhibitor.

6. A pharmaceutical composition for treating arteriosclerosis comprising:
(i) an isolated polypeptide consisting of the sequence (I) Cys-X1-Val-Ala-Pro-X2; and
(ii) an anti-arteriosclerosis agent bound to the polypeptide of (i) as an effective ingredient,
wherein X1 is an amino acid with polar uncharged side chain and X2 is an amino acid with positive charged side chain, and
wherein the anti-arteriosclerosis agent is selected from the group consisting of lovastatin, rapamycin, Celebrex, Ticlopin, Marimastat, and Trocade.

7. A method for drug delivery comprising administering the composition of claim 3 to a subject in need thereof at an effective dose.

8. A method for treating neoplastic disease comprising administering the pharmaceutical composition of claim 3 to a subject in need thereof at an effective dose.

9. A method for treating stroke comprising administering the pharmaceutical composition of claim 4 to a subject in need thereof at an effective dose.

10. A method for treating myocardial infarction comprising administering the pharmaceutical composition of claim 5 to a subject in need thereof at an effective dose.

11. A method for treating arteriosclerosis comprising administering the pharmaceutical composition of claim 6 to a subject in need thereof at an effective dose.

12. A method for imaging a disease site selected from the group consisting of neoplastic disease, stroke, myocardial infarction and arteriosclerosis comprising administering the composition of claim 1 to a subject in need thereof at an effective dose.

13. The composition of claim 1 , wherein the amino acid with polar uncharged side chain is Ser or Thr.

14. The composition of claim 1 , wherein the amino acid with positive charged side chain is Arg or Lys.

15. The composition of claim 1 , wherein the isolated polypeptide is selected from the group consisting of SEQ ID NO: 2 and SEQ ID NOs: 10 to 12.

Assignees

• Lee Byung Heon
• Kim In San
• Kyungpook Nat Univ Ind Acad

Inventors

• Lee Byung Heon
• Kim In San

Classifications

• A61P9/10

  for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis · CPC title

• A61P35/02

  specific for leukemia · CPC title

• A61P35/00

  Antineoplastic agents · CPC title

• A61K49/0032

  Methine dyes, e.g. cyanine dyes · CPC title

• C07K2319/33

  fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies · CPC title