Metabolite analysis (plasma) of [ 18 F]FB-ML5 in a HT1080 xenograft mouse model

In document University of Groningen Design, (radio)synthesis and applications of radiolabelled matrix metalloproteinase inhibitors for PET Matusiak, Nathalie (Page 92-98)

A dual inhibitor of matrix metalloproteinases and a disintegrin and metalloproteinases, [ 18 F]FB-ML5, as a molecular probe for

Scheme 1: Synthesis of the building block 9 Figure 1: Structure and design of ML5

4. Materials and methods 1 General

4.20 Metabolite analysis (plasma) of [ 18 F]FB-ML5 in a HT1080 xenograft mouse model

Metabolite analysis was performed on plasma collected after the ex vivo biodistri-bution study. 750 µL of ACN was added to 250 µL of plasma, the mixture was then centrifuged for 3 min at 3000 rpm. The supernatant was passed through a Millex Filter (0.22 µm), diluted with 600 µL acetonitrile and 600 µL H2O, and analysed by semi-preparative HPLC. Fractions of the eluate were collected every minute and radioactivity in these fractions was then determined with a gammacounter (LKB Wallac, Turku, Finland).

5. Acknowledgements

The authors wish to thank the Dutch Technology Foundation (STW) for financial support (project 08008).

Supplementary information

Molecular modeling of ML5 and FB-ML5

The MolDock Scores for ML5 and FB-ML5 for MMP-2, MMP-9, MMP-12 and ADAM-17 are summarized in [Table 1]. MolDock score is described as the fitness of pose into the binding site by evaluating the intermolecular interaction energy between the ligand and the enzyme, and the intramolecular interaction energy of the enzyme. Docking results of ML5 and FB-ML5 into MMPs shows alignment to the molecule design concept. The substituent construct on ML5 and FB-ML5 which are P1’, P2’ and P3’, correspond with MMPs and ADAM-17 pockets S1’, S2’, and S3’. The hydroxamic acid group was in position to form binding coordination with Zn2+. The detail of the ML5 and FB-ML5 docking results on MMPs and ADAM-17 are described below.

ML5 with MMP-2

The distances between the hydroxamic acid group and Zn2+ are respectively 2.22 and 2.26 Å.

The isopropyl chain is positioned inside the S1’ pocket, whereas the phenyl ring is in the S2’ pocket (S1’ and S2’ are hydrophobic cavities which contain non-polar residues (valine and leucine at S1’; isoleucine, alanine, proline and leucine at S2’).

The lysine side chain of ML5 is solvent exposed.

Several hydrogen bonds are formed with ML5.

FB-ML5 with MMP-2

The distances between the hydroxamic acid group and Zn2+ are respectively 2.15 and 2.27 Å.

The isopropyl chain is positioned inside the S1’ pocket.

Both phenyl and para-chain are solvent exposed. The fluorobenzoyl-lysine cannot adopt the same pose as the fluorobenzoyl-lysine on ML5 since it has a longer chain and is bulky.

The phenyl side chain does not fit into the S2’ pocket, possibly due to the fluoroben-zoyl-lysine side chain which pulls the molecule to be solvent exposed. Moreover, the aromatic interaction between the tyrosine residue and the phenyl ring could contribute to the affinity of FB-ML5.

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ML5 with MMP-9

The distances between the hydroxamic acid group and Zn2+ are respectively 2.11 and 2.26 Å.

The isopropyl chain is positioned inside the S1’ pocket.

The phenyl ring is solvent exposed.

The lysine chain of ML5 fits into the S3’ pocket.

Several hydrogen bonds are formed, which suggests that ML5 can bind tightly to the active site of MMP-9.

FB-ML5 with MMP-9

The distances between the hydroxamic acid group and Zn2+ are respectively 2.10 and 2.18 Å.

The isopropyl chain is positioned inside the S1’ pocket.

The phenyl ring is solvent exposed.

The fluorobenzoyl-lysine side chain fits into the S3’ pocket and is solvent exposed.

ML5 with MMP-12

The lysine side chain of ML5 fits into the S3’ pocket, a little space is left.

FB-ML5 with MMP-12

The fluorobenzoyl-lysine side chain is too large for the S3’ pocket and as a result, the group bends to another direction.

ML5 with ADAM-17

The benzene group is solvent exposed and forms a hydrophobic interaction with isoleucine.

The lysine side chain of ML5 fits nicely into the S3’ pocket and forms hydrogen bonds with alanine.

FB-ML5 with ADAM-17

The fluorobenzoyl-group goes to the S3’ pocket but as it is a big structure, it cannot go deep into this pocket.

The lysine side chain pulls out to the solvent and as a result, less hydrogen bonds can be formed by the backbone of ML5.

References

1. Seals DF, Courtneidge SA. The ADAMs family of metalloproteases: multidomain proteins with mul-tiple functions. Genes Dev. 2003;17(1):7-30.

2. Nagase H, Woessner JF. Matrix metalloproteinases. J Biol Chem. 1999; 274(31):21491-4.

3. White JM. ADAMs: modulators of cell–cell and cell–matrix interactions. Curr Opin Cell Biol.

2003;15(5):598–606.

4. MacFadyen RJ. Can matrix metalloproteinase inhibitors provide a realistic therapy in cardiovascular medicine? Curr Opin Pharmacol. 2007;7(2):171-8.

5. Hu J, Van den Steen PE, Sang Q-XA, Opdenakker G. Matrix metalloproteinase inhibitors as therapy for inflammatory and vascular diseases. Nat Rev Drug Discov. 2007;6:480–98.

6. Hidalgo M, Eckhardt SG. Development of matrix metalloproteinase inhibitors in cancer therapy. J Natl Cancer Inst. 2001;93(3):178–93.

7. Wiley JP, Hughes KA, Kaiser RJ, Kesicki EA, Lund KP, Stolowitz ML. Phenylboronic acid-sali-cylhydroxamic acid bioconjugates. 2. Polyvalent immobilization of protein ligands for affinity chromatography. Bioconjug Chem. 2001;12(2):240–50.

8. Konstantinopoulos PA, Karamouzis MV, Papatsoris AG, Papavassiliou AG. Matrix metalloproteinase inhibitors as anticancer agents. Int J Biochem Cell Biol. 2008;40(6-7):1156-68.

9. Björklund M, Koivunen E. Gelatinase-mediated migration and invasion of cancer cells. Biochim Biophys Acta. 2005;1755(1):37–69.

10. Klein G, Vellenga E, Fraaije MW, Kamps WA, de Bont ESJM. The possible role of matrix metalloprotein-ase (MMP)-2 and MMP-9 in cancer, e.g. acute leukemia. Crit Rev Oncol Hematol. 2004;50(2):87–100.

11. Forsyth PA, Wong H, Laing TD, et al. Gelatinase-A (MMP-2), gelatinase-B (MMP-9) and membrane type matrix metalloproteinase-1 (MT1-MMP) are involved in different aspects of the pathophysiology of malignant gliomas. Br J Cancer. 1999;79:1828–35.

12. Somiari SB, Somiari RI, Heckman CM, et al. Circulating MMP2 and MMP9 in breast cancer - potential role in classification of patients into low risk, high risk, benign disease and breast cancer categories.

Int J Cancer. 2006;119(6):1403–11.

13. Nawrocki B, Polette M, Marchand V, et al. Expression of matrix metalloproteinases and their in-hibitors in human bronchopulmonary carcinomas: quantitative and morphological analyses. Int J Cancer. 1997;72:556–64.

14. Zhou M, Qin S, Chu Y, Wang F, Chen L, Lu Y. Immunolocalization of MMP-2 and MMP-9 in human rheumatoid synovium. Int J Clin Exp Pathol. 2014;7:3048-56.

15. Meijer MJ, Mieremet-Ooms MA, van der Zon AM. Increased mucosal matrix metalloproteinase-1, -2, -3 and -9 activity in patients with inflammatory bowel disease and the relation with Crohn’s disease phenotype. Dig Liver Dis. 2007;39:733-9.

16. Pyke C, Ralfkiær E, Huhtala P, et al. Localization of messenger RNA for Mr 72,000 and 92,000 type IV collagenases in human skin cancers by in situ hybridization. Cancer Res. 1992;52:1336–41.

17. Chandler S, Cossins J, Lury J, Wells G. Macrophage metalloelastase degrades matrix and myelin pro-teins and processes a tumour necrosis factor-alpha fusion protein. Biochem Biophys Res Commun.

1996;228(2):421–9.

18. Dong Z, Kumar R, Yang X, Fidler IJ. Macrophage-derived metalloelastase is responsible for the generation of angiostatin in Lewis lung carcinoma. Cell. 1997;88(6):801–10.

19. Houghton AM, Grisolano JL, Baumann ML, et al. Macrophage elastase (matrix metalloproteinase-12) suppresses growth of lung metastases. Cancer Res. 2006;66(12):6149–55.

CHAPTER

3

20. Black RA, Rauch CT, Kozlosky CJ et al. A metalloproteinase disintegrin that releases tumour-necrosis factor-α from cells. Nature. 1997;385,729-33.

21. Arribas J, Esselens C. ADAM17 as a therapeutic target in multiple diseases. Curr Pharm Des.

2009;15:2319-35.

22. Moss ML, White JM, Lambert MH, Andrews RC. TACE and other ADAM proteases as targets for drug discovery. Drug Discov Today. 2001;6(8):417–26.

23. Jones BA, Riegsecker S, Rahman A, et al. Role of ADAM-17, p38 MAPK, cathepsins, and the protea-some pathway in the synthesis and shedding of fractalkine/CX3CL1 in rheumatoid arthritis. Arthritis Rheum. 2013;65:2814-25.

24. Tursi A, Elisei W, Principi M et al. Mucosal expression of basic fibroblastic growth factor, syndecan 1 and tumour necrosis factor-α in Crohn’s disease in deep remission under treatment with anti-TNFα antibodies. J Gastrointestin Liver Dis. 2014;23:261-5.

25. Kirkegaard T, Pedersen G, Saermark T, Brynskov J. Tumour necrosis factor-alpha converting enzyme (TACE) activity in human colonic epithelial cells. Clin Exp Immunol. 2004;135:146-53.

26. Miller PW, Long NJ, Vilar R, Gee AD. Synthesis of 11C, 18F, 15O, and 13N radiolabels for positron emission tomography. Angew Chem Int Ed Engl. 2008;47(47):8998–9033.

27. Leeuwenburgh MA, Geurink PP, Klein T, et al. Solid-phase synthesis of succinylhydroxamate pep-tides: functionalized matrix metalloproteinase inhibitors. Org Lett. 2006;8:1705–8.

28. Geurink P, Klein T, Leeuwenburgh M, et al. A peptide hydroxamate library for enrichment of metal-loproteinases: towards an affinity-based metalloproteinase profiling protocol. Org Biomol Chem.

2008;6:1244–50.

29. Whittaker M, Floyd CD, Brown P, Gearing AJH. Design and therapeutic application of matrix metal-loproteinase inhibitors. Chem Rev. 1999;99(9):2735-76.

30. Johnström P, Clark JC, Pickard JD, Davenport AP. Automated synthesis of the generic peptide labelling agent N-succinimidyl 4-[18F]fluorobenzoate and application to [18F]-label the vasoactive transmit-ter urotensin-II as a ligand for positron emission tomography. Nucl Med Biol. 2008;35:725–31.

31. Wester HJ, Hamacher K, Stöcklin G. A comparative study of N.C.A. fluorine-18 labeling of proteins via acylation and photochemical conjugation. Nucl Med Biol. 1996;23:365–72.

32. Wüst F, Hultsch C, Bergmann R, Johannsen B, Henle T. Radiolabelling of isopeptide Nɛ-(g-glutamyl)-l-lysine by conjugation with N-succinimidyl-4-[18F]fluorobenzoate. Appl Radiat Isot. 2003;59:43–8.

33. Marik J, Sutcliffe JL. Fully automated preparation of n.c.a. 4-[18F]fluorobenzoic acid and N-succin-imidyl 4-[18F]fluorobenzoate using a Siemens/CTI chemistry process control unit (CPCU). Appl Radiat Isot. 2007;65:199–203.

34. Sounni NE, Devy L, Hajitou A, et al. MT1-MMP expression promotes tumor growth and angiogenesis through an up-regulation of vascular endothelial growth factor expression. FASEB J. 2002;16:555–

64.

35. Köhrmann A, Kammerer U, Kapp M, Dietl J, Anacker J. Expression of matrix metalloproteinases (MMPs) in primary human breast cancer and breast cancer cell lines: New findings and review of the literature. BMC Cancer. 2009;9:188-207.

36. Bremer C, Bredow S, Mahmood U. Optical Imaging of Matrix Metalloproteinase–2 Activity in Tu-mors : Feasibility Study in a Mouse Model. Nat Med. 2001;22:523–9.

37. Bellac CL, Li Y, Lou Y, et al. Novel MMP inhibitor [18F]-Marimastat-aryltrifluoroborate as a probe for in vivo PET imaging in cancer. Cancer Res. 2010;70(19):7562-9.

38. Temma T, Sano K, Kuge Y, et al. Development of a radiolabeled probe for detecting membrane type-1 matrix metalloproteinase on malignant tumors. Biol Pharm Bull. 2009;32(7):1272-7.

39. Shipley JM, Doyle GA, Fliszar CJ, et al. The structural basis for the elastolytic activity of the 92-kDa and 72-kDa gelatinases. J Biol Chem. 1996;271:4335-41.

40. Parkar AA, Stow MD, Smith K, et al. Large-scale expression, refolding, and purification of the catalytic domain of human macrophage metalloelastase (MMP-12) in Escherichia coli. Protein Expr Purif. 2000;20:152-61.

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MicroPET evaluation of a hydroxamate-based MMP inhibitor,

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