Top

European Journal of Nuclear Medicine and Molecular Imaging

Published in:

01-06-2021 | Alzheimer's Disease | Original Article

[¹¹C]MODAG-001—towards a PET tracer targeting α-synuclein aggregates

Authors: Laura Kuebler, Sabrina Buss, Andrei Leonov, Sergey Ryazanov, Felix Schmidt, Andreas Maurer, Daniel Weckbecker, Anne M. Landau, Thea P. Lillethorup, Daniel Bleher, Ran Sing Saw, Bernd J. Pichler, Christian Griesinger, Armin Giese, Kristina Herfert

Published in: European Journal of Nuclear Medicine and Molecular Imaging | Issue 6/2021

Abstract

Purpose

Deposition of misfolded alpha-synuclein (αSYN) aggregates in the human brain is one of the major hallmarks of synucleinopathies. However, a target-specific tracer to detect pathological aggregates of αSYN remains lacking. Here, we report the development of a positron emission tomography (PET) tracer based on anle138b, a compound shown to have therapeutic activity in animal models of neurodegenerative diseases.

Methods

Specificity and selectivity of [³H]MODAG-001 were tested in in vitro binding assays using recombinant fibrils. After carbon-11 radiolabeling, the pharmacokinetic and metabolic profile was determined in mice. Specific binding was quantified in rats, inoculated with αSYN fibrils and using in vitro autoradiography in human brain sections of Lewy body dementia (LBD) cases provided by the Neurobiobank Munich (NBM).

Results

[³H]MODAG-001 revealed a very high affinity towards pure αSYN fibrils (K_d = 0.6 ± 0.1 nM) and only a moderate affinity to hTau46 fibrils (K_d = 19 ± 6.4 nM) as well as amyloid-β_1–42 fibrils (K_d = 20 ± 10 nM). [¹¹C]MODAG-001 showed an excellent ability to penetrate the mouse brain. Metabolic degradation was present, but the stability of the parent compound improved after selective deuteration of the precursor. (d₃)-[¹¹C]MODAG-001 binding was confirmed in fibril-inoculated rat striata using in vivo PET imaging. In vitro autoradiography showed no detectable binding to aggregated αSYN in human brain sections of LBD cases, most likely, because of the low abundance of aggregated αSYN against background protein.

Conclusion

MODAG-001 provides a promising lead structure for future compound development as it combines a high affinity and good selectivity in fibril-binding assays with suitable pharmacokinetics and biodistribution properties.

Available only for authorised users

Spillantini MG, Schmidt ML, Lee VM, Trojanowski JQ, Jakes R, Goedert M. Alpha-synuclein in Lewy bodies. Nature. 1997;388(6645):839–40. https://doi.org/10.1038/42166.CrossRefPubMed

Fanciulli A, Wenning GK. Multiple-system atrophy. N Engl J Med. 2015;372(14):1375–6. https://doi.org/10.1056/NEJMc1501657.CrossRefPubMed

Graham JG, Oppenheimer DR. Orthostatic hypotension and nicotine sensitivity in a case of multiple system atrophy. J Neurol Neurosurg Psychiatry. 1969;32(1):28–34. https://doi.org/10.1136/jnnp.32.1.28.CrossRefPubMedPubMedCentral

Gibb WR, Esiri MM, Lees AJ. Clinical and pathological features of diffuse cortical Lewy body disease (Lewy body dementia). Brain. 1987;110(Pt 5):1131–53. https://doi.org/10.1093/brain/110.5.1131.CrossRefPubMed

Gomez-Tortosa E, Newell K, Irizarry MC, Sanders JL. Hyman BT. alpha-Synuclein immunoreactivity in dementia with Lewy bodies: morphological staging and comparison with ubiquitin immunostaining. Acta Neuropathol. 2000;99(4):352–7. https://doi.org/10.1007/s004010051135.CrossRefPubMed

Lewandowsky MAG, Bumke O. Handbuch der Neurologie. Springer-Verlag Berlin Heidelberg; 1910.

Clark CM, Schneider JA, Bedell BJ, Beach TG, Bilker WB, Mintun MA, et al. Use of florbetapir-PET for imaging beta-amyloid pathology. JAMA. 2011;305(3):275–83. https://doi.org/10.1001/jama.2010.2008.CrossRefPubMedPubMedCentral

Klunk WE, Engler H, Nordberg A, Wang Y, Blomqvist G, Holt DP, et al. Imaging brain amyloid in Alzheimer’s disease with Pittsburgh Compound-B. Ann Neurol. 2004;55(3):306–19. https://doi.org/10.1002/ana.20009.CrossRefPubMed

Villemagne VL, Ong K, Mulligan RS, Holl G, Pejoska S, Jones G, et al. Amyloid imaging with (18)F-florbetaben in Alzheimer disease and other dementias. J Nucl Med. 2011;52(8):1210–7. https://doi.org/10.2967/jnumed.111.089730.CrossRefPubMed

10.

Shahmoradian SH, Lewis AJ, Genoud C, Hench J, Moors TE, Navarro PP, et al. Lewy pathology in Parkinson’s disease consists of crowded organelles and lipid membranes. Nat Neurosci. 2019;22(7):1099–109. https://doi.org/10.1038/s41593-019-0423-2.CrossRefPubMed

11.

Kotzbauer PT, Tu Z, Mach RH. Current status of the development of PET radiotracers for imaging alpha synuclein aggregates in Lewy bodies and Lewy neurites. Clin Transl Imaging. 2017;5(1):3–14. https://doi.org/10.1007/s40336-016-0217-4.CrossRef

12.

Mathis CA, Lopresti BJ, Ikonomovic MD, Klunk WE. Small-molecule PET tracers for imaging proteinopathies. Semin Nucl Med. 2017;47(5):553–75. https://doi.org/10.1053/j.semnuclmed.2017.06.003.CrossRefPubMedPubMedCentral

13.

Pike VW. Considerations in the development of reversibly binding PET radioligands for brain imaging. Curr Med Chem. 2016;23(18):1818–69. https://doi.org/10.2174/0929867323666160418114826.CrossRefPubMedPubMedCentral

14.

Maurer A, Leonov A, Ryazanov S, Herfert K, Kuebler L, Buss S, et al. (11) C Radiolabeling of anle253b: a putative PET tracer for Parkinson’s disease that binds to alpha-synuclein fibrils in vitro and crosses the blood-brain barrier. ChemMedChem. 2019. https://doi.org/10.1002/cmdc.201900689.

15.

Wagner J, Ryazanov S, Leonov A, Levin J, Shi S, Schmidt F, et al. Anle138b: a novel oligomer modulator for disease-modifying therapy of neurodegenerative diseases such as prion and Parkinson’s disease. Acta Neuropathol. 2013;125(6):795–813. https://doi.org/10.1007/s00401-013-1114-9.CrossRefPubMedPubMedCentral

16.

Wegrzynowicz M, Bar-On D, Calo L, Anichtchik O, Iovino M, Xia J, et al. Depopulation of dense alpha-synuclein aggregates is associated with rescue of dopamine neuron dysfunction and death in a new Parkinson’s disease model. Acta Neuropathol. 2019;138(4):575–95. https://doi.org/10.1007/s00401-019-02023-x.CrossRefPubMedPubMedCentral

17.

Heras-Garvin A, Weckbecker D, Ryazanov S, Leonov A, Griesinger C, Giese A, et al. Anle138b modulates alpha-synuclein oligomerization and prevents motor decline and neurodegeneration in a mouse model of multiple system atrophy. Mov Disord. 2019;34(2):255–63. https://doi.org/10.1002/mds.27562.CrossRefPubMed

18.

Reiner AM, Schmidt F, Ryazanov S, Leonov A, Weckbecker D, Deeg AA, et al. Photophysics of diphenyl-pyrazole compounds in solutions and alpha-synuclein aggregates. Biochim Biophys Acta Gen Subj. 2018;1862(4):800–7. https://doi.org/10.1016/j.bbagen.2017.12.007.CrossRefPubMed

19.

Neurobiobank Munich, Germany (NBM). Ludwig Maximilians University Munich [BIORESOURCE]. https://www.neuropathologie.med.uni-muenchen.de/neurobiobank_muenchen/index.html

20.

Auld DS, Farmen MW, Kahl SD, Kriauciunas A, McKnight KL, Montrose C, et al. Receptor Binding Assays for HTS and Drug Discovery. In: Sittampalam GS, Grossman A, Brimacombe K, Arkin M, Auld D, Austin CP, et al., editors. Assay Guidance Manual. Bethesda; 2004.

21.

Zhang X, Jin H, Padakanti PK, Li J, Yang H, Fan J, et al. Radiosynthesis and in vivo evaluation of two PET radioligands for imaging alpha-synuclein. Appl Sci (Basel). 2014;4(1):66–78. https://doi.org/10.3390/app4010066.CrossRef

22.

Pirali T, Serafini M, Cargnin S, Genazzani AA. Applications of deuterium in medicinal chemistry. J Med Chem. 2019;62(11):5276–97. https://doi.org/10.1021/acs.jmedchem.8b01808.CrossRefPubMed

23.

Ishikawa M, Hashimoto Y. Improvement in aqueous solubility in small molecule drug discovery programs by disruption of molecular planarity and symmetry. J Med Chem. 2011;54(6):1539–54. https://doi.org/10.1021/jm101356p.CrossRefPubMed

24.

Bagchi DP, Yu L, Perlmutter JS, Xu J, Mach RH, Tu Z, et al. Binding of the radioligand SIL23 to alpha-synuclein fibrils in Parkinson disease brain tissue establishes feasibility and screening approaches for developing a Parkinson disease imaging agent. PLoS One. 2013;8(2):e55031. https://doi.org/10.1371/journal.pone.0055031.CrossRefPubMedPubMedCentral

25.

Chu W, Zhou D, Gaba V, Liu J, Li S, Peng X, et al. Design, synthesis, and characterization of 3-(Benzylidene)indolin-2-one derivatives as ligands for alpha-synuclein fibrils. J Med Chem. 2015;58(15):6002–17. https://doi.org/10.1021/acs.jmedchem.5b00571.CrossRefPubMedPubMedCentral

26.

Fodero-Tavoletti MT, Mulligan RS, Okamura N, Furumoto S, Rowe CC, Kudo Y, et al. In vitro characterisation of BF227 binding to alpha-synuclein/Lewy bodies. Eur J Pharmacol. 2009;617(1–3):54–8. https://doi.org/10.1016/j.ejphar.2009.06.042.CrossRefPubMed

27.

Hsieh CJ, Xu K, Lee I, Graham TJA, Tu Z, Dhavale D, et al. Chalcones and five-membered heterocyclic isosteres bind to alpha synuclein fibrils in vitro. ACS Omega. 2018;3(4):4486–93. https://doi.org/10.1021/acsomega.7b01897.CrossRefPubMedPubMedCentral

28.

Kikuchi A, Takeda A, Okamura N, Tashiro M, Hasegawa T, Furumoto S, et al. In vivo visualization of alpha-synuclein deposition by carbon-11-labelled 2-[2-(2-dimethylaminothiazol-5-yl)ethenyl]-6-[2-(fluoro)ethoxy]benzoxazole positron emission tomography in multiple system atrophy. Brain. 2010;133(Pt 6):1772–8. https://doi.org/10.1093/brain/awq091.CrossRefPubMed

29.

Verdurand M, Levigoureux E, Lancelot S, Zeinyeh W, Billard T, Quadrio I, et al. Amyloid-beta radiotracer [(18)F]BF-227 does not bind to cytoplasmic glial inclusions of postmortem multiple system atrophy brain tissue. Contrast Media Mol Imaging. 2018;2018:9165458. https://doi.org/10.1155/2018/9165458.CrossRefPubMedPubMedCentral

30.

Verdurand M, Levigoureux E, Zeinyeh W, Berthier L, Mendjel-Herda M, Cadarossanesaib F, et al. In silico, in vitro, and in vivo evaluation of new candidates for alpha-synuclein PET imaging. Mol Pharm. 2018;15(8):3153–66. https://doi.org/10.1021/acs.molpharmaceut.8b00229.CrossRefPubMed

31.

Yue X, Dhavale DD, Li J, Luo Z, Liu J, Yang H, et al. Design, synthesis, and in vitro evaluation of quinolinyl analogues for alpha-synuclein aggregation. Bioorg Med Chem Lett. 2018;28(6):1011–9. https://doi.org/10.1016/j.bmcl.2018.02.031.CrossRefPubMedPubMedCentral

32.

Celej MS, Jares-Erijman EA, Jovin TM. Fluorescent N-arylaminonaphthalene sulfonate probes for amyloid aggregation of alpha-synuclein. Biophys J. 2008;94(12):4867–79. https://doi.org/10.1529/biophysj.107.125211.CrossRefPubMedPubMedCentral

33.

Volkova KD, Kovalska VB, Balanda AO, Losytskyy MY, Golub AG, Vermeij RJ, et al. Specific fluorescent detection of fibrillar alpha-synuclein using mono- and trimethine cyanine dyes. Bioorg Med Chem. 2008;16(3):1452–9. https://doi.org/10.1016/j.bmc.2007.10.051.CrossRefPubMed

34.

Zhang L, Villalobos A, Beck EM, Bocan T, Chappie TA, Chen L, et al. Design and selection parameters to accelerate the discovery of novel central nervous system positron emission tomography (PET) ligands and their application in the development of a novel phosphodiesterase 2A PET ligand. J Med Chem. 2013;56(11):4568–79. https://doi.org/10.1021/jm400312y.CrossRefPubMed

35.

Strome EM, Jivan S, Doudet DJ. Quantitative in vitro phosphor imaging using [3H] and [18F] radioligands: the effects of chronic desipramine treatment on serotonin 5-HT2 receptors. J Neurosci Methods. 2005;141(1):143–54. https://doi.org/10.1016/j.jneumeth.2004.06.008.CrossRefPubMed

36.

Sihver W, Sihver S, Bergstrom M, Murata T, Matsumura K, Onoe H, et al. Methodological aspects for in vitro characterization of receptor binding using 11C-labeled receptor ligands: a detailed study with the benzodiazepine receptor antagonist [11C]Ro 15–1788. Nucl Med Biol. 1997;24(8):723–31. https://doi.org/10.1016/s0969-8051(97)00113-3.CrossRefPubMed

37.

Iwai A, Masliah E, Yoshimoto M, Ge N, Flanagan L, de Silva HA, et al. The precursor protein of non-A beta component of Alzheimer’s disease amyloid is a presynaptic protein of the central nervous system. Neuron. 1995;14(2):467–75. https://doi.org/10.1016/0896-6273(95)90302-x.CrossRefPubMed

38.

Masliah E, Iwai A, Mallory M, Ueda K, Saitoh T. Altered presynaptic protein NACP is associated with plaque formation and neurodegeneration in Alzheimer’s disease. Am J Pathol. 1996;148(1):201–10.PubMedPubMedCentral

39.

Twohig D, Nielsen HM. Alpha-synuclein in the pathophysiology of Alzheimer’s disease. Mol Neurodegener. 2019;14(1):23. https://doi.org/10.1186/s13024-019-0320-x.CrossRefPubMedPubMedCentral

40.

Ueda K, Fukushima H, Masliah E, Xia Y, Iwai A, Yoshimoto M, et al. Molecular cloning of cDNA encoding an unrecognized component of amyloid in Alzheimer disease. Proc Natl Acad Sci U S A. 1993;90(23):11282–6. https://doi.org/10.1073/pnas.90.23.11282.CrossRefPubMedPubMedCentral

41.

Strohaker T, Jung BC, Liou SH, Fernandez CO, Riedel D, Becker S, et al. Structural heterogeneity of alpha-synuclein fibrils amplified from patient brain extracts. Nat Commun. 2019;10(1):5535. https://doi.org/10.1038/s41467-019-13,564-w.CrossRefPubMedPubMedCentral

42.

Falcon B, Zhang W, Murzin AG, Murshudov G, Garringer HJ, Vidal R, et al. Structures of filaments from Pick’s disease reveal a novel tau protein fold. Nature. 2018;561(7721):137–40. https://doi.org/10.1038/s41586-018-0454-y.CrossRefPubMedPubMedCentral

43.

Falcon B, Zivanov J, Zhang W, Murzin AG, Garringer HJ, Vidal R, et al. Novel tau filament fold in chronic traumatic encephalopathy encloses hydrophobic molecules. Nature. 2019;568(7752):420–3. https://doi.org/10.1038/s41586-019-1026-5.CrossRefPubMedPubMedCentral

44.

Fitzpatrick AWP, Falcon B, He S, Murzin AG, Murshudov G, Garringer HJ, et al. Cryo-EM structures of tau filaments from Alzheimer’s disease. Nature. 2017;547(7662):185–90. https://doi.org/10.1038/nature23002.CrossRefPubMedPubMedCentral

45.

Zhang W, Falcon B, Murzin AG, Fan J, Crowther RA, Goedert M, et al. Heparin-induced tau filaments are polymorphic and differ from those in Alzheimer’s and Pick’s diseases. Elife. 2019;8. https://doi.org/10.7554/eLife.43584.

46.

Bergeron C, Pollanen M. Lewy bodies in Alzheimer disease—one or two diseases? Alzheimer Dis Assoc Disord. 1989;3(4):197–204.CrossRefPubMed

47.

Hamilton RL. Lewy bodies in Alzheimer’s disease: a neuropathological review of 145 cases using alpha-synuclein immunohistochemistry. Brain Pathol. 2000;10(3):378–84.CrossRefPubMed

48.

Colom-Cadena M, Gelpi E, Charif S, Belbin O, Blesa R, Marti MJ, et al. Confluence of alpha-synuclein, tau, and beta-amyloid pathologies in dementia with Lewy bodies. J Neuropathol Exp Neurol. 2013;72(12):1203–12. https://doi.org/10.1097/Nen.0000000000000018.CrossRefPubMed

49.

Irwin DJ, Lee VM, Trojanowski JQ. Parkinson’s disease dementia: convergence of alpha-synuclein, tau and amyloid-beta pathologies. Nat Rev Neurosci. 2013;14(9):626–36. https://doi.org/10.1038/nrn3549.CrossRefPubMedPubMedCentral

50.

Robinson JL, Lee EB, Xie SX, Rennert L, Suh E, Bredenberg C, et al. Neurodegenerative disease concomitant proteinopathies are prevalent, age-related and APOE4-associated. Brain. 2018;141:2181–93. https://doi.org/10.1093/brain/awy146.CrossRefPubMedPubMedCentral

51.

Flores-Fernandez JM, Rathod V, Wille H. Comparing the folds of prions and other pathogenic amyloids. Pathogens. 2018;7(2). https://doi.org/10.3390/pathogens7020050.

52.

Harada R, Okamura N, Furumoto S, Yanai K. Imaging protein misfolding in the brain using beta-sheet ligands. Front Neurosci. 2018;12:585. https://doi.org/10.3389/fnins.2018.00585.CrossRefPubMedPubMedCentral

53.

Serpell LC, Berriman J, Jakes R, Goedert M, Crowther RA. Fiber diffraction of synthetic alpha-synuclein filaments shows amyloid-like cross-beta conformation. Proc Natl Acad Sci U S A. 2000;97(9):4897–902. https://doi.org/10.1073/pnas.97.9.4897.CrossRefPubMedPubMedCentral

54.

Hsieh CJ, Ferrie JJ, Xu K, Lee I, Graham TJA, Tu Z, et al. Alpha synuclein fibrils contain multiple binding sites for small molecules. ACS Chem Neurosci. 2018;9(11):2521–7. https://doi.org/10.1021/acschemneuro.8b00177.CrossRefPubMed

55.

Breydo L, Uversky VN. Structural, morphological, and functional diversity of amyloid oligomers. FEBS Lett. 2015;589(19 Pt A):2640–8. https://doi.org/10.1016/j.febslet.2015.07.013.CrossRefPubMed

Title: [11C]MODAG-001—towards a PET tracer targeting α-synuclein aggregates
Authors: Laura Kuebler
Sabrina Buss
Andrei Leonov
Sergey Ryazanov
Felix Schmidt
Andreas Maurer
Daniel Weckbecker
Anne M. Landau
Thea P. Lillethorup
Daniel Bleher
Ran Sing Saw
Bernd J. Pichler
Christian Griesinger
Armin Giese
Kristina Herfert
Publication date: 01-06-2021
Publisher: Springer Berlin Heidelberg
Keywords: Alzheimer's Disease
Positron Emission Tomography
Alzheimer's Disease
Parkinson's Disease
Published in: European Journal of Nuclear Medicine and Molecular Imaging / Issue 6/2021
Print ISSN: 1619-7070
Electronic ISSN: 1619-7089
DOI: https://doi.org/10.1007/s00259-020-05133-x

Keynote webinar | Spotlight on medication adherence

Springer Medicine

[¹¹C]MODAG-001—towards a PET tracer targeting α-synuclein aggregates

Abstract

Purpose

Methods

Results

Conclusion

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Purpose

Methods

Results

Conclusion

Please log in to get access to this content

Other articles of this Issue 6/2021

Interim [18F]FDG PET/CT can predict response to anti-PD-1 treatment in metastatic melanoma

PSMA-PET: is the time to say goodbye to metabolic radiopharmaceuticals in prostate cancer?

Correction to: [18F]FDG -PET/CT uptake reveals a single metastasis inside vertebral hemangioma, catch me if you scan

Biodistribution, pharmacokinetics, dosimetry of [68Ga]Ga-DOTA.SA.FAPi, and the head-to-head comparison with [18F]F-FDG PET/CT in patients with various cancers

Diagnostic value of [18F]FDG-PET/CT in polymyalgia rheumatica: a systematic review and meta-analysis

Correction to: 68Ga-FAPI outperforms 18F-FDG PET/CT in identifying solitary fibrous tumor