SYNTHESIS OF LIMITED COMPLEXING AGENTS

Claims

1. A method for preparing a compound of formula (I): Formula (I), its diastereomer, its salt, its solvate, or a combination of any of the foregoing, wherein the method includes: (a) step (1-1) comprising reacting a compound of formula (Ia): Formula (Ia) or its diastereomer with a compound of formula (Ib): Formula (Ib) in a solvent and in the presence of a reducing agent to form a compound of formula (Ib): Formula (Iв) or its diastereomer; (b) step (1-2), comprising reacting a compound of formula (Ib) or a salt thereof with a compound of formula (Id): Formula (Ig) in the presence of a base and in a solvent with the formation of a compound of formula (Id): Formula (Id) or its diastereomer; (c) step (1-3), comprising reacting the compound of formula (Id) with a base in a solvent to form a compound of formula (Ie): Formula (Ie) or its diastereomer; (g) step (1-4), comprising reacting the compound of formula (Ie) with the compound of formula (Izh): Formula (Iж) in a solvent and in the presence of a carboxyl activating reagent to form a compound of formula (Ii): Formula (Ii) or its diastereomer; and (d) step (1-5) comprising reacting a compound of formula (Ii) with an acid to form a compound of formula (I), a diastereomer thereof, a salt thereof, or a solvate thereof, or a combination of any of the foregoing, Where each of R 1 and R 2 independently represents C1-C6 alkyl; R 3 is a halogen; and each of R 4 , R 5 , R 6 and R 7 independently represents halogen or hydrogen.

2. The method according to claim 1, wherein the solvent in step (1-1) comprises alcohol.

3. The method according to claim 2, wherein the solvent in step (1-1) is ethanol.

4. The method according to any one of claims 1-3, wherein the reducing agent in step (1-1) is a hydride.

5. The method according to claim 4, wherein the reducing agent in step (1-1) is NaBH(OAc)3.

6. The method according to any one of claims 1-5, wherein step (1-1) further comprises purifying the compound of formula (Ib) using column chromatography.

7. The method according to any one of claims 1-6, wherein the solvent in step (1-2) is a polar organic solvent.

8. The method according to claim 7, wherein the solvent in step (1-2) is CH3CN.

9. The method according to any one of claims 1-8, wherein the base in step (1-2) is an organic base.

10. The method of claim 9, wherein the base in step (1-2) is an amine base.

11. The method of claim 10, wherein the base in step (1-2) is N,N-diisopropylethylamine (DIPEA).

12. The method according to any one of claims 1-11, wherein step (1-2) further comprises purifying the compound of formula (Id) using column chromatography.

13. The method according to any one of claims 1-12, wherein the solvent in step (1-3) is an aqueous solvent.

14. The method according to claim 13, wherein the solvent in step (1-3) further comprises tetrahydrofuran (THF).

15. The method according to any one of paragraphs 1-14, wherein the base in step (1-3) is KOH.

16. The method according to any one of claims 1-15, wherein the solvent in step (1-4) is a halogenated hydrocarbon.

17. The method of claim 16, wherein the solvent in step (1-4) is CH2Cl2.

18. The method according to any one of claims 1 to 17, wherein the carboxyl activating reagent is 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI).

19. The method according to any one of claims 1-18, wherein the acid in step (1-5) is trifluoroacetic acid.

20. The method according to any one of claims 1-19, wherein step (1-5) further comprises purifying the compound of formula (Ii) using preparative high performance liquid chromatography.

21. The method according to any one of paragraphs 1-20, in which R 1is methyl.

22. The method according to any one of paragraphs 1-21, in which R 2 is tert-butyl.

23. The method according to any one of paragraphs 1-22, in which R 3 represents Br.

24. The method according to any one of paragraphs 1-23, in which each of R 4 , R 5 , R 6 and R 7 represents F.

25. The method according to any one of claims 1 to 24, wherein the compound of formula (I) is compound 1: Connection 1.

26. A method for preparing a compound of formula (I): Formula (I), its diastereomer, its salt or its solvate, or a combination of any of the foregoing, wherein the method includes: (a) step (2-1), comprising reacting a compound of formula (Ia): Formula (Ia) with a compound of formula (Ib): Formula (Ib) in a solvent and in the presence of a reducing agent to form a compound of formula (Ib): Formula (Iв); (b) step (2-2), comprising reacting the compound of formula (Ib) with an acid to obtain a salt of formula (Ib); (c) step (2-3), which involves reacting a salt of formula (Ic) with a compound of formula (Id): Formula (Ig) in the presence of a base and in a solvent with the formation of a compound of formula (Id): Formula (Id), its diastereomer or its salt; (d) step (2-4), comprising reacting the compound of formula (Id) with a base in a solvent to form a compound of formula (2e): Formula (Ie), its diastereomer or its salt; and (d) step (2-5), comprising reacting a compound of formula (Ie) or a salt thereof with a compound of formula (Izh): Formula (Iж) in a solvent and in the presence of a carboxyl activating reagent to form a compound of formula (Ii): Formula (Ii), its diastereomer or its salt; and (e) step (2-6) comprising reacting the compound of formula (Ii) with an acid to form a compound of formula (I), a diastereomer thereof, a salt thereof, a solvate thereof, or a combination of any of the foregoing, Where each of R 1 and R 2 independently represents C1-C6 alkyl; R 3 is a halogen; and each of R 4 , R 5 , R 6and R 7 independently represents halogen or hydrogen.

27. The method according to claim 26, wherein the solvent in step (2-1) comprises alcohol.

28. The method according to claim 27, wherein the solvent in step (2-1) is ethanol.

29. The method according to any one of paragraphs 26-28, wherein the reducing agent in step (2-1) comprises a hydride.

30. The method of claim 29, wherein the reducing agent in step (2-1) is NaBH(OAc)3.

31. The method according to any one of claims 26-30, wherein the acid in step (2-2) is orotic acid.

32. The method according to any one of paragraphs 26-31, in which in step (2-2) a salt of formula (Ib) is obtained having the following structure: Orotate of formula (Iв).

33. The method according to claim 32, wherein step (2-2) comprises crystallization of the salt of formula (Ib) or its diastereomer in the form of an orotate salt.

34. The method according to any one of claims 26-30, wherein the acid in step (2-2) is fumaric acid.

35. The method according to any one of paragraphs 26-30 or 34, in which in step (2-2) a salt of formula (Ib) is obtained having the following structure: Fumarate of formula (Ib).

36. The method according to claim 34 or 35, wherein step (2-2) further comprises crystallization of the salt of formula (Ib) or its diastereomer in the form of a fumarate salt.

37. The method according to any one of paragraphs 26-36, wherein the base in step (2-3) is K2CO3.

38. The method according to any one of paragraphs 26-37, wherein the solvent in step (2-3) comprises toluene.

39. The method according to any one of paragraphs 26-38, wherein the solvent in step (2-3) comprises water.

40. The method according to any one of paragraphs 26-39, wherein the base in step (2-4) is KOH.

41. The method according to any one of paragraphs 26-40, wherein the solvent in step (2-4) comprises THF.

42. The method according to any one of paragraphs 26-41, wherein the solvent in step (2-4) comprises water.

43. The method according to any one of claims 26 to 42, wherein step (2-4) comprises obtaining a salt of formula (Ie) in the form of an HCl salt, and wherein step (2-4) further comprises crystallizing the salt of formula (Ie).

44. The method according to any one of paragraphs 26-43, wherein the solvent in step (2-5) is a halogenated hydrocarbon.

45. The method of claim 44, wherein the solvent in step (2-5) is CH2Cl2.

46. ​​The method according to any one of paragraphs 26-45, wherein the carboxyl activating reagent in step (2-5) is EDCI.

47. The method according to any one of paragraphs 26-46, wherein the solvent in step (2-6) is dioxane.

48. The method according to any one of paragraphs 26-47, wherein the acid in step (2-6) is HCl.

49. The method according to any one of paragraphs 26-48, wherein in step (2-6) a salt of formula (I) or a solvate thereof is obtained having the following structure: Formula (I-1) HCl.

50. The method according to claim 49, wherein step (2-6) further comprises crystallizing the salt of formula (I).

51. The method according to any one of paragraphs 26-50, in which R 1 is methyl.

52. The method according to any one of paragraphs 26-51, in which R 2 is tert-butyl.

53. The method according to any one of paragraphs 26-52, in which R 3 represents Br.

54. The method according to any one of paragraphs 26-50, in which R 4 , R 5 , R 6 and R 7 represent F.

55. The method according to any one of claims 26-54, wherein the salt of the solvate of formula (I) has the structure: Connection 2.

56. A composition comprising one or more diastereomers of a compound of formula (I), its salts or solvates thereof: Formula (I), obtained using the method according to any of paragraphs 1-25.

57. The composition of claim 56, wherein the composition comprises at least about 90% (w / w) of the trans-diastereomer of the compound of formula (I).

58. The composition of claim 57, wherein the trans-diastereomer has the structure: Formula (I-1).

59. The composition according to any one of paragraphs 56-58, in which R 4 , R 5 , R 6 and R 7 represent F.

60. A salt of formula (I) or a solvate thereof, wherein the salt of formula (I) has the structure: Formula (I-1) HCl, obtained using the method according to any of paragraphs 26-55.

61. Salt according to claim 60, in which R4 , R 5 , R 6 and R 7 represent F.

62. Compound of formula (Id): Formula (Id), its diastereomer or its salt, where each of R 1 and R 2 independently represents C1-C6 alkyl.

63. Compound of formula (Ib): Formula (Iв), its diastereomer or its salt, where R 1 is C1-C6 alkyl.

64. A composition comprising a complex comprising a compound obtained by the method according to any one of claims 1-55 and a metal halide.

65. The composition of claim 64, wherein the metal halide is {Al 18 F} 2+ .

66. The composition of claim 65, wherein at least 90% (w / w) of the compound in the composition has the following structure: Formula (I-1).

67. The composition according to claim 66, in which R 4 , R 5 , R 6 and R 7 represent F.

68. The composition according to any one of claims 64-67, further comprising a polypeptide, wherein the complex is conjugated to the polypeptide.

69. The composition of claim 68, wherein the polypeptide is an antibody or antibody fragment.

70. The composition of claim 69, wherein the antibody fragment is selected from the group consisting of VHH, scFv, minibody, Fab, Fab' and Fv fragments.

71. The composition of claim 68, wherein the polypeptide is a framework protein other than an antibody.

72. The composition of claim 71, wherein the non-antibody scaffold protein is selected from the group consisting of affibody, affilin, knottin, fibronectin, anticalin, atrimer, avimer, FN3 scaffold, finomer, Kunitz domain, pronectin, OBody, and DARPin.

73. The composition of any one of claims 64-72, wherein the composition is a pharmaceutical composition suitable for administration to a subject.

74. The composition of claim 73, wherein the pharmaceutical composition is suitable for intravenous administration.

75. The composition according to claim 73 or 74, wherein the subject is a human being.

76. A method for obtaining a labeled agent containing a polypeptide and 18 F radionuclide, including: (a) contacting the polypeptide with a compound obtained using the method of any one of claims 1 to 55 under conditions that allow conjugation of the complex with the polypeptide to form a conjugate; and (b) contacting the conjugate with an aluminum fluoride complex containing 18 F, with the production of a labeled agent.

77. The method of claim 76, wherein the polypeptide is an antibody or antibody fragment.

78. The method of claim 77, wherein the antibody fragment is selected from the group consisting of VHH, scFv, minibody, Fab, Fab' and Fv fragments.

79. The method of claim 76, wherein the polypeptide is a framework protein other than an antibody.

80. The method of claim 79, wherein the non-antibody scaffold protein is selected from the group consisting of affibody, affilin, knottin, fibronectin, anticalin, atrimer, avimer, FN3 scaffold, finomer, Kunitz domain, pronectin, OBody, and DARPin.

81. Crystalline salt of a compound with the structure: Orotate of compound 1b, wherein the salt has at least one of the following properties: (a) a powder X-ray diffraction pattern showing characteristic peaks at approximately 7.5° 2-theta, approximately 13.9° 2-theta, approximately 16.6° 2-theta, approximately 23.4° 2-theta, and approximately 25.6° 2-theta; (b) essentially the same powder X-ray diffraction pattern as the "orotate type A" in Fig. 1; (c) thermogravimetric analysis thermogram showing at 20-150°C a mass loss of approximately 3.3%; (d) essentially the same thermogravimetric analysis thermogram as shown in Fig. 2; (d) differential scanning calorimetry thermogram showing an endothermic peak at approximately 172.8°C; (e) essentially the same differential scanning calorimetry thermogram as shown in Fig. 2; (g) spectrum 1 H nuclear magnetic resonance with a characteristic peak at approximately 5.68 ppm; or (and) essentially the same spectrum 1 H nuclear magnetic resonance as shown in Fig.

3.

82. Crystalline salt of a compound with the structure: Orotate of compound 1b, wherein the salt has at least one of the following properties: (a) a powder X-ray diffraction pattern showing characteristic peaks at approximately 7.2° 2-theta, approximately 14.0° 2-theta, approximately 15.2° 2-theta, approximately 22.7° 2-theta, and approximately 25.0° 2-theta; (b) essentially the same powder X-ray diffraction pattern as the "orotate type B" in Fig. 1; (c) thermogravimetric analysis thermogram showing a mass loss of approximately 2.6% at 20-150°C; (d) essentially the same thermogravimetric analysis thermogram as shown in Fig. 4; (d) differential scanning calorimetry thermogram showing an exothermic peak at approximately 161.7 °C and an endothermic peak at approximately 182.9 °C; (e) essentially the same differential scanning calorimetry thermogram as shown in Fig. 4; (g) spectrum 1H nuclear magnetic resonance with a characteristic peak at approximately 5.69 ppm; and (and) essentially the same spectrum 1 H nuclear magnetic resonance as shown in Fig.

5.

83. Crystalline salt of a compound with the structure: Orotate of compound 1b, wherein the salt has at least one of the following properties: (a) a powder X-ray diffraction pattern with characteristic peaks at approximately 3° 2-theta, approximately 7.8° 2-theta, approximately 9.7° 2-theta, approximately 13.9° 2-theta, approximately 16.7° 2-theta, approximately 21.4° 2-theta, approximately 23.4° 2-theta; (b) essentially the same powder X-ray diffraction pattern as the "wet sample" or "dry sample" as shown in Fig. 6; (c) thermogravimetric analysis thermogram showing a mass loss of approximately 4.8% at 20-150°C; (d) essentially the same thermogravimetric analysis thermogram as shown in Fig. 7; (d) a differential scanning calorimetry thermogram showing an exothermic peak at approximately 178.1 °C and an endothermic peak at approximately 185.6 °C; (e) essentially the same differential scanning calorimetry thermogram as shown in Fig. 7; (g) spectrum 1 H nuclear magnetic resonance with a characteristic peak at approximately 5.68 ppm; and (and) essentially the same spectrum 1 H nuclear magnetic resonance as shown in Fig.

8.

84. Crystallized salt of a compound with the structure: Fumarate of compound 1b, wherein the salt has at least one of the following properties: (a) a powder X-ray diffraction pattern showing characteristic peaks at approximately 7.9° 2-theta, approximately 15.8° 2-theta, approximately 18.6° 2-theta, approximately 19.3° 2-theta, approximately 20.1° 2-theta, approximately 21.0° 2-theta, and approximately 24.7° 2-theta; (b) essentially the same powder X-ray diffraction pattern as the "wet sample" or "dry sample" as shown in Fig. 9; (c) thermogravimetric analysis thermogram showing a mass loss of approximately 2.4% at 20-140°C; (d) essentially the same thermogravimetric analysis thermogram as shown in Fig. 10; (d) differential scanning calorimetry thermogram showing an endothermic peak at approximately 154.7°C; (e) essentially the same differential scanning calorimetry thermogram as shown in Fig. 10; (g) spectrum 1H nuclear magnetic resonance with a characteristic peak at approximately 6.39 ppm; and (and) essentially the same spectrum 1 H nuclear magnetic resonance as shown in Fig.

11.

85. Crystallized solvate of a salt of a compound with the structure: Compound 2, wherein the solvate of the salt has at least one of the following properties: (a) a powder X-ray diffraction pattern with characteristic peaks at approximately 6.8° 2-theta, approximately 17.6° 2-theta, approximately 20.3° 2-theta, approximately 22.2° 2-theta, and approximately 23.3° 2-theta; and (b) essentially the same powder X-ray diffraction pattern as shown in Fig.

15.

86. A limited complexing agent containing: (i) a compound obtained using the method of any one of paragraphs 1-55; or (ii) a crystalline salt or solvate according to any one of claims 81 to 85.

87. A label fragment comprising a detectable label comprising a metal halide conjugated or chelated to a linker, comprising: (i) a compound obtained using the method of any one of paragraphs 1-55; or (ii) a crystalline salt or solvate according to any one of claims 81 to 85.

88. A label-conjugated marker-binding protein comprising a detectable label comprising a metal halide linked to said marker-binding protein via a linker, comprising: (i) a compound obtained using the method of any one of paragraphs 1-55; or (ii) a crystalline salt or solvate according to any one of claims 81 to 85.

89. A method for visualizing a molecule, said method comprising exposing said molecule to a label-conjugated marker-binding protein containing a detectable label containing a metal halide linked to said marker-binding protein via a linker containing (i) a compound obtained using the method of any one of paragraphs 1-55; or (ii) a crystalline salt or solvate according to any one of paragraphs 81 to 85, wherein said marker-binding protein binds said molecule; and tag detection.