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METHOD OF PREPARING ESTRADIOL DERIVATIVES BY SOLID-PHASE SYNTHESIS

20240116978 ยท 2024-04-11

    Inventors

    Cpc classification

    International classification

    Abstract

    The present invention relates to a method of preparing estradiol derivatives and/or estrone derivatives, which are suitable for radiolabeling. The present invention further relates to the estradiol derivatives and/or estrone derivatives, preferably obtained by the method of the present invention, as well as to the use of the estradiol derivatives and/or estrone derivatives for radiolabeling with diagnostic and/or therapeutic radionuclides. The present invention further relates to a method of imaging and/or diagnosis of breast cancer as well as to a method of treatment of breast cancer.

    Claims

    1. A method of preparing an estradiol derivative and/or estrone derivative for radiolabeling, comprising the steps of (1) preparing an acetic acid-estradiol precursor; (2) coupling the acetic acid-estradiol precursor obtained in step (1) to a solid phase synthesis resin, wherein said resin is suitable for Fmoc (9-fluorenylmethyl-oxycarbonyl)-solid phase synthesis and carries Fmoc-lysine with a protecting group at the lysine side chain; (3) removing the protecting group at the lysine side chain; (4) coupling a bifunctional group for radiolabeling to the lysine side chain; and (5) removal of the estradiol derivative and/or the estrone derivative from the resin.

    2. The method of claim 1, wherein step (1) comprises reacting estrone and tert-butyl bromoacetate.

    3. The method of claim 2, wherein in step (1) estrone and tert-butyl bromoacetate are reacted in the presence of potassium carbonate (K.sub.2CO.sub.3) and potassium iodide (KI).

    4. The method of claim 2, wherein at the end of step (1) the tert-butyl protecting group is removed from the acetic acid-estradiol precursor.

    5. The method of claim 1, wherein the solid phase synthesis resin is Wang resin.

    6. The method of claim 1, wherein the protecting group at the Lys side chain resin is ivDde (4,4-dimethyl-2,6-dioxocyclohex-1-ylidene)-3-methylbutyl-) and the resin is Fmoc-Lys(ivDde)-Wang resin.

    7. The method of claim 1, wherein before the coupling of step (2), the acetic acid-estradiol precursor is preactivated at its ?-carboxylic function by using an activating reagent.

    8. The method of claim 1, wherein said activating reagent is HBTU (O-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate) which is used in the presence of a base.

    9. The method of claim 8, wherein the base is DIEA (diisopropylethylamine).

    10. The method of claim 1, wherein the bifunctional group for radiolabeling is selected from DOTA (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid), NOTA (1,4,7-triazacyclononane-1,4,7-triacetic acid) and AoA (aminooxy acetic acid).

    11. The method of claim 1, wherein in step (4) for coupling the bifunctional group for radiolabeling to the Lys side chain DOTA-tris-(t-Bu ester), NOTA-bis-(t-Bu ester) or Boc-AoA is used, wherein said DOTA-tris-(t-Bu ester), NOTA-bis-(t-Bu ester) or Boc-AoA is preactivated.

    12. The method of claim 11, wherein the DOTA-tris-(t-Bu ester), NOTA-bis-(t-Bu ester) or Boc-AoA is preactivated with HBTU and DIEA.

    13. The method of claim 1, wherein at least one of steps (1) to (5) is carried out at a temperature in the range from about 18 to about 30? C.

    14. The method of claim 1, wherein all steps are carried out at a temperature in the range from about 18 to about 30? C.

    15. The method of claim 1, further comprising the step of (6) radiolabeling the estradiol derivative and/or estrone derivative obtained in step (5) with one or more diagnostic and/or therapeutic radionuclides, and/or with one or more diagnostic radiotracers.

    16. The method of claim 15, wherein the one or more diagnostic and/or therapeutic radionuclides are selected from .sup.68Ga, .sup.177Lu, .sup.64Cu, .sup.89Zr, .sup.18F, .sup.18FDG and .sup.18F-FBA (fluorobenzaldehyde).

    17. The method of claim 15, wherein the diagnostic radiotracer is .sup.18FDG (fluorodeoxyglucose).

    18. An estradiol derivative or an estrone derivative selected from DOTA-estradiol, NOTA-estradiol, AoA-estradiol, DOTA-estrone, NOTA-estrone and AoA-estrone.

    19. The estradiol derivative or the estrone derivative of claim 18, which is selected from DOTA-estradiol-lysine, NOTA-estradiol-lysine, AoA-estradiol-lysine, DOTA-estrone-lysine, NOTA-estrone-lysine and AoA-estrone-lysine.

    20. The estradiol derivative or the estrone derivative of claim 18, which further comprises one or more diagnostic and/or therapeutic radionuclides, and/or one or more diagnostic radiotracers.

    21. The estradiol derivative or the estrone derivative of claim 20, wherein the one or more diagnostic and/or therapeutic radionuclides are selected from .sup.68Ga, .sup.177Lu, .sup.64Cu, .sup.89Zr, .sup.18F, .sup.18FDG and .sup.18F-FBA (fluorobenzaldehyde).

    22. The estradiol derivative or the estrone derivative of claim 20, wherein the diagnostic radiotracer is .sup.18FDG (fluorodeoxyglucose).

    23. A method of imaging and/or diagnosis of breast cancer, comprising the step of: administering a diagnostic amount of an estradiol derivative or an estrone derivative of claim 18 to a subject.

    24. The method of claim 23, wherein the breast cancer is estrogen-receptor-positive breast cancer.

    25. A method of imaging and/or diagnosis of breast cancer, comprising the step of: administering a diagnostic amount of an estradiol derivative or an estrone derivative of claim 18 to a subject, wherein the estradiol derivative or the estrone derivative further comprises one or more diagnostic radionuclides or one or more diagnostic radiotracers.

    26. A method of treatment of breast cancer, comprising the step of: administering a therapeutic amount of an estradiol derivative or an estrone derivative of claim 18 to a subject in need thereof.

    27. The method of claim 26, wherein the breast cancer is estrogen-receptor-positive breast cancer.

    28. A method of treatment of breast cancer, comprising the step of: administering a therapeutic amount of an estradiol derivative or an estrone derivative of claim 18 to a subject in need thereof, wherein the estradiol derivative or the estrone derivative further comprises one or more therapeutic radionuclides.

    Description

    BRIEF DESCRIPTION OF THE DRAWINGS

    [0191] FIG. 1. .sup.18F-fluoroestradiol (.sup.18F-FES)

    [0192] FIG. 2. Scheme of the preparation of 3-acetic acid estradiol. [0193] (a) K.sub.2CO.sub.3, KI, DMF, 23? C., 24 h; [0194] (b) TFA, DCM, 23? C., 2 h.

    [0195] FIGS. 3A-3C. Scheme of the preparation of DOTA/NOTA/AoA-estrone as well as DOTA/NOTA/AoA-estradiol by solid phase synthesis. [0196] (3A) and (3B) Scheme of the preparation of DOTA/NOTA/AoA-estrone by solid phase synthesis. [0197] (3C) DOTA/NOTA/AoA-estradiol derivatives.

    [0198] FIGS. 4A-4D. Mass spectrometry analysis of the precursor and the estradiol derivatives. [0199] (4A) Mass spectrometry of 3-acetic acid estradiol. [0200] LC-MS: m/z calcd for C.sub.20H.sub.24O.sub.4 328.4; found, 328.8 [M+H].sup.+. [0201] (4B) Mass spectrometry of DOTA-estradiol. [0202] LC-MS: m/z calcd for C.sub.42H.sub.63N.sub.6O.sub.12 843.5; found, 844.5 [M+H].sup.+ [0203] (4C) Mass spectrometry of NOTA-estradiol. [0204] LC-MS: m/z calcd for C.sub.38H.sub.59N.sub.6O.sub.9 743.5; found, 744.6 [M+H].sup.+. [0205] (4D) Mass spectrometry of AoA-estradiol. [0206] LC-MS: m/z calcd for C.sub.28H.sub.40N.sub.3O.sub.7 530.3; found, 531.7 [M+H].sup.+.

    EXAMPLES

    1. Materials and Methods

    [0207] All chemicals, solvents and reagents for the synthesis of estradiol derivatives were procured from different commercial sources and used without further purification. .sup.177Lu radionuclide and .sup.68Ge/.sup.68Ga generator were bought from ITG Isotope Technologies Garching GmbH, Germany.

    [0208] .sup.18FDG, .sup.64Cu and .sup.89Zr radionuclides are produced at on-site facility of the Cyclotron and Radiopharmaceuticals Department.

    [0209] Solid-phase synthesis of the estradiol derivatives was started by utilizing the commercially available Fmoc-Lys(ivDde)-wang resin using the peptide synthesis glass reaction vessel (Peptides International, Louisville, USA).

    [0210] The structures of the newly synthesized DOTA/NOTA/AoA-estradiol derivatives were confirmed by mass spectrometry analyses performed on Agilent 6125 single quadrupole liquid chromatography/mass spectrometry system (LC/MS) (Agilent Technologies, Santa Clara, CA, USA) using an eluent of 0.1% formic acid/29.95% water/69.95% acetonitrile at a flow rate of 0.3 mL/min.

    [0211] The features disclosed in the foregoing description, in the claims and/or in the accompanying drawings may, both separately and in any combination thereof, be material for realizing the invention in diverse forms thereof.

    REFERENCES

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