DIAGNOSTIC AGENT

• US 20100284934A1
• Filed: 08/29/2008
• Published: 11/11/2010
• Est. Priority Date: 08/30/2007
• Status: Active Grant

• Current Assignee: United Arab Emirates University (Al Hikma Development Co. PJSC)
• Original Assignee: United Arab Emirates University (Al Hikma Development Co. PJSC)
• Inventors: El-Agnaf, Omar
• Granted Patent: US 8,613,905 B2
• Time in Patent Office: Days
• US Class Current: 424/9.34
• CPC Class Codes: A61K 49/085 conjugated systems
• CPC Class Codes: A61K 49/14 Peptides, e.g. proteins
• CPC Class Codes: A61K 49/146 the peptide being a polyamino acid, e.g. poly-lysine
• CPC Class Codes: A61P 25/16 Anti-Parkinson drugs
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: C07K 14/47 from mammals
• CPC Class Codes: G01N 2800/2814 Dementia; Cognitive disorders
• CPC Class Codes: G01N 2800/2821 Alzheimer
• CPC Class Codes: G01N 2800/2835 Movement disorders, e.g. Parkinson, Huntington, Tourette
• CPC Class Codes: G01N 2800/52 Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis
• CPC Class Codes: G01N 33/6896 Neurological disorders, e.g. Alzheimer's disease

Abstract

The invention provides an agent comprising an amino acid sequence for use in a method of diagnosis of a synucleinopathic disease.

Claims

1. An agent for use in a diagnostic method practised on the human or animal body wherein the agent comprises a peptide comprising the amino acid sequence of from two to twelve contiguous amino acid residues from the sequence corresponding to amino acid residues 61 to 95 of SEQ ID NO:

1, or comprises a derivative or analogue of said amino acid sequence, wherein the agent binds to α

-synuclein aggregates with a higher affinity than to free α

-synuclein.

2. An agent according to claim 1, wherein the amino acid sequence comprises a maximum of seven contiguous amino acid residues, a maximum of six contiguous amino acid residues, a maximum of five contiguous amino acid residues, a maximum of four contiguous amino acid residues, a maximum of three contiguous amino acids, or a maximum of contiguous two amino acid residues from amino acid residues 61 to 95 of SEQ ID NO:

1.

3. An agent according to claim 1, wherein the amino acid sequence comprises the sequence of SEQ ID NO:

3, SEQ ID NO;

4, SEQ ID NO;

5, SEQ ID NO;

6 or SEQ ID NO;

7.

4. An agent according to claim 1 comprising a derivative or analogue of the amino acid sequence as defined in claim 1.

5. An agent according to claim 4, wherein the derivative is a D-amino acid derivative, an N-methylated amino acid derivative, or a peptoid analogue.

6. An agent according to claim 4, wherein the derivative or analogue is more protease-resistant than the peptide from which it is derived.

7. An agent according to claim 4, wherein the derivative or analogue is adapted to facilitate its entry into biological cells, or across biological barriers.

8. An agent according to claim 7, wherein the derivative is adapted by the addition of a membrane-permeable carrier peptide selected from HIV-1 Tat-(48-60), flock house virus (FHV) coat-(35-49), Drosophila Antennapedia-(43-58), basic peptides or polyamines.

9. An agent according to claim 1 further comprising a detectable label or a substituent capable of complexing to a detectable label.

10. An agent according to claim 9, wherein the detectable label is a contrast agent.

11. An agent according to claim 10, wherein the contrast agent comprises gadolinium ions.

12. An agent according to claim 9, wherein the substituent capable of complexing to a detectable label is 1,4,7,10-tetraazacyclododecane-1,4,7-tris(acetic acid-t-butyl ester)-10-acetic acid (DOTA).

13. An agent according to claim 9 wherein the detectable label, or the substituent capable of complexing to a detectable label, is present at the C-terminus and/or at the N-terminus.

14. An agent as defined in claim 1 for use in the diagnosis of a synucleinopathic disease involving α

-synuclein and/or fragment(s) of α

-synuclein.

19. The agent of claim 14 wherein the synucleinopathic disease is Parkinson'"'"'s Disease (PD), Alzheimer'"'"'s Disease (AD), dementia with Lewy bodies (DLB) or multiple system atrophy (MSA).

20. Use of an agent as defined in claim 9 in a method of imaging α

-synuclein aggregates.

21. A method of imaging α

-synuclein aggregates, said method comprising detecting the binding of an agent as defined in claim 9 to α

-synuclein aggregates.

22. A method according to claim 21, wherein the α

-synuclein aggregates are present in a human or animal subject.

23. A method according to claim 22, further comprising administering an agent as defined in claim 9 to said subject.

24. A method according to claim 21, wherein said agent is detected by magnetic resonance imaging (MRI)

25. A method of diagnosing a synucleinopathic disease involving α

-synuclein and/or fragment(s) of α

-synuclein, said method comprising administering an agent as defined in claim 1 to a subject and thereby detecting the presence or absence of α

-synuclein aggregates, wherein the presence of α

-synuclein aggregates indicates that the subject has a synucleinopathic disease and the absence of α

-synuclein aggregates indicates that the subject does not have said synucleinopathic disease.

26. A method of monitoring a synucleinopathic disease involving α

-synuclein and/or fragment(s) of α

-synuclein, said method comprising administering an agent as defined in claim 1 to a subject and detecting the amount and/or size of any α

-synuclein aggregates that have been formed by α

-synuclein alone or by α

-synuclein complexed with any other protein(s).

27. A method according to claim 26, wherein said subject is undergoing therapy to treat said synucleinopathic disease and said method is for monitoring the effectiveness of said therapy.

28. The method of claim 25 wherein the synucleinopathic disease is Parkinson'"'"'s Disease (PD), Alzheimer'"'"'s Disease (AD), dementia with Lewy bodies (DLB) or multiple system atrophy (MSA).

29. A method according to claim 25, wherein the agent is administered by non-surgical means.

30. A kit for imaging α

-synuclein aggregates, said kit comprising an agent according to claim 1 and means for administering the agent to a subject.

31. An in vitro method of diagnosing a synucleinopathic disease involving α

-synuclein and/or fragment(s) of α

-synuclein in a patient comprising;

(a) combining a sample of tissue and/or biological fluid (e.g. blood, CSF, urine) from the patient with an agent according to claim 9 for a time and under conditions effective to allow binding of the agent to aggregates of α

-synuclein present in the tissue sample; and

(b) thereby detecting the presence or absence of aggregates of α

-synuclein in the sample, wherein the presence of α

-synuclein aggregates indicates that the subject has a synucleinopathic disease and the absence of α

-synuclein aggregates indicates that the subject does not have said synucleinopathic disease.

32. The method of claim 31 further comprising:

(c) quantifying the amount of agent bound to aggregates of α

-synuclein in the sample.

33. The method of claim 32 further comprising:

(d) combining a later-acquired sample of tissue from the patient with an amount of a agent for a time and under conditions effective to allow binding of the agent aggregates of α

-synuclein present in the later-acquired sample;

(e) quantifying the amount of agent bound to aggregates of α

-synuclein in the later-acquired sample.

(f) comparing the quantity of α

-synuclein in the sample of step (c) to the quantity of α

-synuclein in the sample of step (e); and

(g) thereby determining whether the condition of the patient has altered, wherein a higher quantity of α

-synuclein in the sample of step (e) than in the sample of step (c) indicates that the condition of the patient has worsened, and wherein a higher quantity of α

-synuclein in the sample of step (c) than in the sample of step (e) indicates that the condition of the patient has improved.

34. The method of claim 31, wherein the label of the agent comprises a radiolabel, an enzyme label, a fluorescent label, a chemiluminescent label, or an antigen label.

35. The method of claim 34 wherein the presence of the agent bound to soluble and/or aggregates of α

-synuclein in the sample is detected by autoradiography, positron emission tomography, nuclear magnetic resonance imaging, a gamma counter, or a scintillation counter.

15. A peptide comprising:

i) the amino acid sequence DThr-DVal-DVal-DAla or DVal-DVal-DAla;

ii) a poly-D-Arginine peptide linked to the N- or C-terminus of the sequence of (i) by a Glycine or N-methlyglycine residue and/or any other spacer; and

iii) the substituent DOTA linked to the N-terminus of the peptide.

16. A peptide according to claim 15 wherein the DOTA ligand is complexed with gadolinium ions.

17. A peptide according to claim 15 represented by any one of formulae (I) to (VI):

(I) DOTA-DArg-Nmethylglycine-DVal-DVal-DAla-N-methylglycine-poly-D-Arginine[r₆]

(II) DOTA-DArg-Nmethylglycine-DThr-DVal-DVal-DAla Nmethylglycine-poly-D-Arginine[r₆]

(III) DOTA-DArg-Glyeine-DVal-DVal-DAla-Glycine-poly-D-Arginine[r₆]

(IV) DOTA-DArg-Glycine-DThr-DVal-DVal-DAla-Glycine-poly-D-Arginine[r₆]

(V) DOTA-poly-D-Arginine[r₆]-Glyeine-DVal-DVal-DAla-Glycine-DArg

(VI) DOTA-poly-D-Arginine[r₆]-Glycine-DThr-DVal-DVal-DAla-Glycine-DArg

(VII) DOTA-DArg-Nmethylglycine-DVal-DVal-DAla-N-methylglycine-butadiamine

(VIII) DOTA-DArg-Nmethylglycine-DVal-DVal-DAla-N-methylglycine-pentadiamine

18. A peptide as defined in claim 15 for use in the diagnosis of a synucleinopathic disease involving α

-synuclein and/or fragment(s) of α

-synuclein.

39. The peptide of claim 18 wherein the synucleinopathic disease is Parkinson'"'"'s Disease (PD), Alzheimer'"'"'s Disease (AD), dementia with Lewy bodies (DLB) or multiple system atrophy (MSA).

36. A method of monitoring the effectiveness of a therapeutic agent that has been administered for the purpose of treating a synucleinopathic disease involving α

-synuclein and/or fragment(s) of α

-synuclein, the method comprising;

analysing in vitro a sample from an animal model for the presence and amount of aggregates of α

-synuclein;

or

imaging the brain of an animal model for the presence and amount of α

-synuclein aggregates.

37. (canceled)

38. (canceled)