# US 8,772,226 B2 · Methods and compositions for the detection of cancer

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- Number: US 8,772,226 B2
- Document ID: US8772226B2
- Status: Granted
- Kind: B2
- Application number: 13/257,131
- Filed: 2010-03-17
- Published: 2012-04-19
- Granted: 2014-07-08
- Earliest claimed: 2009-03-17
- Assignees: The Johns Hopkins University
- Applicants: Samuel Ray Denmeade; John Tod Isaacs; Soeren Brogger Christensen
- Inventors: Samuel Ray Denmeade; John Tod Isaacs; Soeren Brogger Christensen
- Web page: https://patentgrep.com/patent/US8772226B2

## CPC

- A61K 31/343: condensed with a carbocyclic ring, e.g. coumaran, bufuralol, befunolol, clobenfurol, amiodarone
- A61P 35/00: Antineoplastic agents
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS

## Abstract

Various embodiments of this invention relate generally to targeted activation and delivery of therapeutic drugs to cells that produce prostate specific antigen (PSA), prostate specific membrane antigen (PSMA) or human glandular kallikrein (hK2). Various further embodiments relate more specifically to PSMA-specific peptide prodrugs that become activated to yield therapeutic drugs. Further aspects of various embodiments of the present invention also relate to methods and compositions for treating or preventing cancers and methods and compositions for detecting and/or imaging cancers.

## Claims

### Claim 1

A composition comprising:
a) thapsigargin (TG) or a thapsigargin analog;
b) a phenolic linker; and
c) a peptide cleavable by a protein selected from:
i) prostate specific membrane antigen (PSMA);
ii) prostate specific antigen (PSA); and
iii) human glandular kallikrein 2 (hK2)
wherein said phenolic linker is conjugated to said thapsigargin or thapsigargin analogue.

### Claim 2 (depends on 1)

The composition of claim 1, wherein the phenolic linker further comprises a radiolabel.

### Claim 3 (depends on 2)

The composition of claim 2, wherein the radiolabel is at least one of 125I, 124I, 131I, or 3H.

### Claim 4 (depends on 1)

The composition of claim 1, wherein the peptide is cleavable by PSMA.

### Claim 5 (depends on 4)

The composition of claim 4, wherein the composition has the following chemical structure:
wherein X1 can be present or absent and when present is selected from the group consisting of 124I, 125I, 131I, and 3H.

### Claim 6 (depends on 4)

The composition of claim 4, wherein the peptide comprises the sequence Asp-Glu*Glu*Glu*Glu (SEQ ID NO:57).

### Claim 7 (depends on 4)

The composition of claim 4, wherein the peptide consists of Asp-Glu*Glu*Glu*Glu (SEQ ID NO:57).

### Claim 8 (depends on 4)

The composition of claim 4, wherein the peptide comprises Asp-Glu.

### Claim 9 (depends on 4)

The composition of claim 4, wherein the peptide consists of Asp-Glu.

### Claim 10 (depends on 1)

The composition of claim 1, wherein the peptide is cleavable by PSA.

### Claim 11 (depends on 10)

The composition of claim 10, wherein the composition has the following chemical structure:
wherein X1 can be present or absent and when present is selected from the group consisting of 124I, 125I, 131I, and 3H.

### Claim 12 (depends on 10)

The composition of claim 10, wherein the peptide is selected from the group consisting of Ser-Lys-Leu-Gln-Leu (SEQ ID NO:42), Ile-Ser-Tyr-Gln-Leu (SEQ ID NO:43), Lys-Ser-Lys-Gln-Leu (SEQ ID NO:44), Ser-Ser-Lys-Leu-Gln-Leu (SEQ ID NO:45), Lys-Ile-Ser-Tyr-Gln-Leu (SEQ ID NO:46), Thr-Lys-Ser-Lys-Gln-Leu (SEQ ID NO:47), His-Ser-Ser-Lys-Leu-Gln-Leu (SEQ ID NO:48), Asn-Lys-Ile-Ser-Tyr-Gln-Leu (SEQ ID NO:49), Ala-Thr-Lys-Ser-Lys-Gln-Leu (SEQ ID NO:50), Glu-His-Ser-Ser-Lys-Leu-Gln-Leu (SEQ ID NO:51), Gln-Asn-Lys-Ile-Ser-Tyr-Gln-Leu (SEQ ID NO:52), Glu-Asn-Lys-Ile-Ser-Tyr-Gln-Leu (SEQ ID NO:53), Ala-Thr-Lys-Ser-Lys-Gln-His-Leu (SEQ ID NO: 55), and His-Ser-Ser-Lys-Leu-Gln-Leu (SEQ ID NO:56).

### Claim 13 (depends on 12)

The composition of claim 12, wherein the peptide consists of His-Ser-Ser-Lys-Leu-Gln-Leu (SEQ ID NO:56).

### Claim 14 (depends on 12)

The composition of claim 12, wherein the peptide further comprises a capping group attached to the N-terminus of the peptide, the group inhibiting endopeptidase activity.

### Claim 15 (depends on 14)

The composition of claim 14, wherein the capping group is selected from the group consisting of acetyl, morpholinocarbonyl, benzyloxycarbonyl, glutaryl, and succinyl substituents.

### Claim 16 (depends on 1)

The composition of claim 1, wherein the peptide is cleavable by hK2.

### Claim 17 (depends on 16)

The composition of claim 16, wherein the peptide is selected from the group consisting of Lys-Arg-Arg (SEQ ID NO:1), Ser-Arg-Arg (SEQ ID NO:2), Ala-Arg-Arg (SEQ ID NO:3), His-Arg-Arg (SEQ ID NO:4), Gln-Arg-Arg (SEQ ID NO:5), Ala-Phe-Arg (SEQ ID NO:6), Ala-Gln-Arg (SEQ ID NO:7), Ala-Lys-Arg (SEQ ID NO:8), Ala-Arg-Lys (SEQ ID NO:9), Ala-His-Arg (SEQ ID NO:10), Gln-Lys-Arg-Arg (SEQ ID NO:11), Lys-Ser-Arg-Arg (SEQ ID NO:12), Ala-Lys-Arg-Arg (SEQ ID NO:13), Lys-Lys-Arg-Arg (SEQ ID NO:14), His-Lys-Arg-Arg (SEQ ID NO:15), Lys-Ala-Phe-Arg (SEQ ID NO:16), Lys-Ala-Gln-Arg (SEQ ID NO:17), Lys-Ala-Lys-Arg (SEQ ID NO:18), Lys-Ala-Arg-Lys (SEQ ID NO:19), Lys-Ala-His-Arg (SEQ ID NO:20), Lys-Arg-Arg-Leu (SEQ ID NO:21), Ser-Arg-Arg-Leu (SEQ ID NO:22), Ala-Arg-Arg-Leu (SEQ ID NO:23), Ala-Arg-Arg-Ser (SEQ ID NO:24), His-Arg-Arg-Ala (SEQ ID NO:25), Gln-Arg-Arg-Leu (SEQ ID NO:26), Ala-Phe-Arg-Leu (SEQ ID NO:27), Ala-Gln-Arg-Leu (SEQ ID NO:28), Ala-Lys-Arg-Leu (SEQ ID NO:29), Ala-Arg-Lys-Leu (SEQ ID NO:30), Ala-His-Arg-Leu (SEQ ID NO:31), His-Ala-Gln-Lys-Arg-Arg-Leu (SEQ ID NO:32), Gly-Gly-Lys-Ser-Arg-Arg-Leu (SEQ ID NO:33), His-Glu-Gln-Lys-Arg-Arg-Leu (SEQ ID NO:34), His-Glu-Ala-Lys-Arg-Arg-Leu (SEQ ID NO:35), Gly-Gly-Gln-Lys-Arg-Arg-Leu (SEQ ID NO:36), His-Glu-Gln-Lys-Arg-Arg-Ala (SEQ ID NO:37), Gly-Gly-Ala-Lys-Arg-Arg-Leu (SEQ ID NO:38), His-Glu-Gln-Lys-Arg-Arg-Ser (SEQ ID NO:39), Gly-Gly-Lys-Lys-Arg-Arg-Leu (SEQ ID NO:40), Gly-Gly-His-Lys-Arg-Arg-Leu (SEQ ID NO:41) and Gly-Gly-Lys-Ala-Arg-Arg-Leu (SEQ ID NO:54).

### Claim 18 (depends on 17)

The composition of claim 17, wherein the peptide consists of Gly-Gly-Lys-Ala-Arg-Arg-Leu (SEQ ID NO:54).

### Claim 19 (depends on 17)

The composition of claim 17, wherein the peptide further comprises a capping group attached to the N-terminus of the peptide, the group inhibiting endopeptidase activity.

### Claim 20 (depends on 19)

The composition of claim 19, wherein the capping group is selected from the group consisting of acetyl, morpholinocarbonyl, benzyloxycarbonyl, glutaryl, and succinyl substituents.

### Claim 21 (depends on 13)

The composition of claim 13, wherein the peptide further comprises a capping group attached to the N-terminus of the peptide, the group inhibiting endopeptidase activity.

### Claim 22 (depends on 18)

The composition of claim 18, wherein the peptide further comprises a capping group attached to the N-terminus of the peptide, the group inhibiting endopeptidase activity.

### Claim 23

A composition for detecting a prostate cancer comprising:
a) thapsigargin (TG) or a thapsigargin analog;
b) a phenolic linker comprising a radiolabel; and
c) a peptide cleavable by a protein selected from:
i) prostate specific membrane antigen (PSMA);
ii) prostate specific antigen (PSA); and
iii) human glandular kallikrein 2 (hK2)
wherein said phenolic linker is conjugated to said thapsigargin or thapsigargin analogue.

### Claim 24 (depends on 23)

The composition of claim 23, wherein said radiolabel is at least one of 125I, 124I, 131I, or 3H.

### Claim 25

A composition for detecting a cancer comprising:
a) thapsigargin (TG) or a thapsigargin analog;
b) a phenolic linker comprising a radiolabel; and
c) a peptide cleavable by prostate specific membrane antigen (PSMA),
wherein said phenolic linker is conjugated to said thapsigargin or thapsigargin analogue.

### Claim 26 (depends on 25)

The composition of claim 25, wherein said radiolabel is at least one of 125I, 124I, 131I, or 3H.

### Claim 27

A method of imaging or treating a subject having cancer or suspected of having cancer comprising administering to the subject a composition comprising:
a) thapsigargin (TG) or a thapsigargin analog;
b) a phenolic linker comprising a radiolabel; and
c) a peptide cleavable by prostate specific membrane antigen (PSMA);
wherein said phenolic linker is conjugated to said thapsigargin or thapsigargin analogue.

### Claim 28 (depends on 27)

The method of claim 27, wherein the imaging the subject further comprises single photon emission computed tomography (SPECT).

### Claim 29 (depends on 28)

The method of claim 28, wherein the radiolabel is 125I.

### Claim 30 (depends on 27)

The method of claim 27, wherein the imaging the subject further comprises positron emission tomography (PET).

### Claim 31 (depends on 30)

The method of claim 30, wherein the radiolabel is 124I.

### Claim 32 (depends on 27)

The method of claim 27, wherein the treating the subject further comprises combination drug and radiation therapy.

### Claim 33 (depends on 32)

The method of claim 32, wherein the radiolabel is 131I.

### Claim 34 (depends on 27)

The method of claim 27, wherein the cancer is at least one of prostate cancer, breast cancer, renal cancer, colon cancer or transitional cell carcinomas.

### Claim 35 (depends on 27)

The method of claim 27, wherein the radiolabel is 3H.

### Claim 36 (depends on 27)

The method of claim 27, wherein the composition has a chemical structure selected from the group consisting of:
wherein X1 is selected from the group consisting of 124I, 125I, 131I, and 3H.

### Claim 37

A method of imaging or treating a subject having prostate cancer or suspected of having prostate cancer comprising administering to the subject a composition comprising:
a) thapsigargin (TG) or a thapsigargin analog;
b) a phenolic linker comprising a radiolabel; and
c) a peptide cleavable by a protein selected from:
i) prostate specific membrane antigen (PSMA);
ii) prostate specific antigen (PSA); and
iii) human glandular kallikrein 2 (hK2)
wherein said phenolic linker is conjugated to said thapsigargin or thapsigargin analogue.

### Claim 38 (depends on 37)

The method of claim 37, wherein the imaging the subject further comprises single photon emission computed tomography (SPECT).

### Claim 39 (depends on 38)

The method of claim 38, wherein the radiolabel is 125I.

### Claim 40 (depends on 37)

The method of claim 37, wherein the imaging the subject further comprises positron emission tomography (PET).

### Claim 41 (depends on 40)

The method of claim 40, wherein the radiolabel is 124I.

### Claim 42 (depends on 37)

The method of claim 37, wherein the treating the subject further comprises combination drug and radiation therapy.

### Claim 43 (depends on 40)

The method of claim 40, wherein the radiolabel is 131I.

### Claim 44 (depends on 37)

The method of claim 37, wherein the radiolabel is 3H.

### Claim 45 (depends on 37)

The method of claim 37, wherein the composition has a chemical structure selected from the group consisting of:
wherein X1 is selected from the group consisting of 124I, 125I, 131I, and 3H.