Dense star polymer conjugates

• US 5,527,524 A
• Filed: 04/05/1993
• Issued: 06/18/1996
• Est. Priority Date: 08/18/1986
• Status: Expired due to Term

• Current Assignee: Donald A. Tomalia, AVC Holdings Inc. (Dow, Inc.), Dendritic NanoTechnologies, Inc. (Loveland Products, Inc.)
• Original Assignee: The Dow Chemical Company (Dow, Inc.)
• Inventors: Edwards, David S., Kruper, William J. Jr., Fazio, Michael J., Tomlinson, Ian A., Jung, Chu W., Tomalia, Donald A., Wilson, Larry R., Hedstrand, David M., Cheng, Roberta C., Kaplan, Donald A.
• Primary Examiner(s): Page, Thurman K.
• Application Number:
  US08/043,198
• Time in Patent Office: 1,170 Days
• Field of Search: 436/531, 436/532, 436/533, 436/535, 424/78.17, 424/78.18, 424/78.19, 424/78.22, 424/78.23, 424/78.26, 424/78.27, 424/78.34, 424/78.37, 424/84, 424/93.1, 424/94.2, 424/184.1 , 424/130.1, 424/401, 424/487, 424/486, 424/409, 424/407, 424/405, 424/452, 424/484, 424/84, 424/1.1, 424/1.21, 424/2, 424/4, 424/9, 424/417, 525/DIG. 16, 514/772.1, 514/772.3, 514/772.4, 514/772.5, 514/772.6, 514/772.7, 523/1.05
• US Class Current: 424/1.33
• CPC Class Codes: A01N 25/10 Macromolecular compounds
• CPC Class Codes: A61K 47/59 obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes
• CPC Class Codes: A61K 47/595 Polyamides, e.g. nylon polyamino acids A61K47/62
• CPC Class Codes: A61K 47/6885 the conjugate or the polymer being a starburst, a dendrimer, a cascade
• CPC Class Codes: A61K 47/6949 inclusion complexes, e.g. clathrates, cavitates or fullerenes
• CPC Class Codes: B82Y 5/00 Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
• CPC Class Codes: C07C 211/29 the carbon skeleton being further substituted by halogen atoms or by nitro or nitroso groups
• CPC Class Codes: C07C 233/11 with carbon atoms of carboxamide groups bound to carbon atoms of an unsaturated carbon skeleton containing six-membered aromatic rings
• CPC Class Codes: C07C 237/20 the carbon skeleton containing six-membered aromatic rings
• CPC Class Codes: C08G 73/02 Polyamines
• CPC Class Codes: C08G 83/003 Dendrimers
• CPC Class Codes: C08L 101/005 Dendritic macromolecules
• CPC Class Codes: C12N 15/87 Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
• CPC Class Codes: Y10S 424/16 Dendrimers and dendritic polymers

Abstract

Dense star polymer conjugates which are composed of at least one dendrimer in association with at least one unit of a carried agricultural, pharmaceutical, or other material have been prepared. These conjugates have particularly advantageous properties due to the unique characteristics of the dendrimer.

Claims

1. A dense star polymer conjugate which comprises at least one dense star polymer of at least generation 3.0 associated with at least one unit of at least one carried material, other than a dye or dye moiety, with the proviso that the carried material maintains its effectiveness in the conjugate.

2. The conjugate of claim 1 wherein the dense star polymer is a dendrimer.

3. The conjugate of claim 2 wherein the dendrimer is radially symmetrical.

4. A conjugate of claim 1 for use as a diagnostic agent.

5. A conjugate of claim 2 for use as a diagnostic agent.

6. The conjugate of claim 1 wherein at least one of the carried materials is a radionuclide, chelator, chelated metal, signal generator, signal reflector, signal absorber, or fragrance.

7. The conjugate of claim 6 wherein at least one of the carried materials is a signal generator, signal reflector or signal absorber.

8. The conjugate of claim 7 for use as a reagent in positron emmission tomography, computer aided tomography, or magnetic resonance imaging.

9. The conjugate of claim 7 wherein at least one of the carried materials is a metal ion, paramagnetic or magnetic entity, fluorescence entity, phosphorescence entity, UV absorber, or electron beam opacifier.

10. The conjugate of claim 7 wherein the signal absorber is a contrast agent.

11. The conjugate of claim 10 wherein the contrast agent is prepared by reaction of an α

,β

-unsaturated ester, an α

,β

-unsaturated ketone, an α

,β

-unsaturated nitrile, an α

,β

-unsaturated carboxyl, isothiocyanato or epoxide moiety.

12. The conjugate of claim 11 wherein the carried material is hydrophobic.

13. The conjugate of claim 11 wherein the carried material is hydrophilic.

14. The conjugate of claim 10 wherein the contrast agent is derived from 1,4,7,10-tetraazacyclododecane-N,N'"'"',N"-triacetic acid.

15. The conjugate of claim 14 wherein the dendrimer portion of the conjugate is a polyamidoamine.

16. The conjugate of claim 14 wherein the dendrimer portion of the conjugate is a polyethylenimine.

17. The conjugate of claim 10 wherein the contrast agent is derived from diethylenetriaminepentaacetic acid.

18. The conjugate of claim 10 wherein the contrast agent comprises a metal ion selected from Gd, Mn and Fe.

19. The conjugate of claim 18 wherein the dendrimer portion of the conjugate is a polyamidoamine.

20. The conjugate of claim 7 wherein the signal generator is a fluorescence agent and the dendrimer portion is a polyethyleneimine.

21. The conjugate of claim 7 wherein the signal generator is fluoresceine and the dendrimer portion is a polyamidoamine.

22. The conjugate of claim 7 wherein the signal generator is a radionuclide.

23. The conjugate of claim 22 wherein the radionuclide is ⁹⁰ Y and the dendrimer portion is a polyamidoamine.

24. The conjugate of claim 6 wherein the signal reflector is a metal ion selected from Fe, Rh, Pd, and Y.

25. The conjugate of claim 1 wherein the dendrimer contains discontinuities.

60. A process for preparing a conjugate of claim 1 wherein the dense star polymer is a dendrimer and the carried material is a lanthanide which comprises chelation of the lanthanide with a starburst polyethyleneimine acetate.

61. The conjugate of claim 2 wherein the dendrimer is of the formula ##EQU10## wherein:

the (Core) is an initiator molecule or atom;

Terminal Moiety is the atom or molecule which occupies the binding sites of the Repeat Unit;

G is the number of generations;

N_r is the multiplicity of the Repeat Unit;

N_c is the multiplicity of the Core molecule or core atom; and

the Repeat Unit has a valency or functionality of N_r +1 wherein N_r is as defined before.

62. The conjugate of claim 2 wherein the dendrimer is of the formula ##EQU11## wherein i is 1 to t-1;

(I) is the initiator core compound, molecule or atom;

(Z^c) represents the binding sites available on the core (I);

N_c represents the core functionality or number of binding sites;

Xⁱ Yⁱ (Zⁱ)_N.spsb.i wherein "i" represents the particular generation from the first to the t-1 generation;

each Z¹ of the first generation repeating unit is connected to an X² of a repeating unit of the second generation and so on through the generations such that each Zⁱ group for a repeating unit Xⁱ Yⁱ (Zⁱ)_N.spsb.i in generation number "i" is connected to the tail (Xⁱ⁺¹) of the repeating unit of the generation number "i+1"; and

X^t Y^t (Z^t)_N.spsb.t wherein t represents a terminal unit and X^t, Y^t, Z^t and N^t may be the same as or different from Xⁱ, Yⁱ, Zⁱ and Nⁱ except that there is no succeeding generation connected to the Z^t groups and N^t may be less than two;

the II function is the product of all the values between its defined limits, such as ##EQU12## which is the number of repeat units, Xⁱ Yⁱ (Zⁱ)_N.spsb.i, comprising the ith generation of one dendritic branch and when i is 1, then ##EQU13##

113. The conjugate of claim 1 wherein the dendrimer is solvent soluble.

26. A dense star polymer conjugate of the formula:

space="preserve" listing-type="equation">(P).sub.x *(M).sub.y (I)

wherein each P represents a dendrimer of at least generation 3.0;

x represents an integer of 1 or greater;

each M represents a unit of a carried material, other than a dye or dye moiety, said carried material can be the same carried material or a different carried material, with the proviso that the carried material maintains its effectiveness in the conjugate;

y represents an integer of 1 or greater; and

* indicates that the carried material is associated with the dendrimer.

27. The conjugate of claim 26 wherein M is a radionuclide, chelator, chelated metal, signal generator, signal reflector, signal absorber or fragrance.

28. The conjugate of claim 26 wherein x=1 and y=2 or more.

29. The conjugate of claim 26 wherein the molar ratio of any ionic M to P is 0.1-1,000:

1.

30. The conjugate of claim 26 wherein M is a signal generator, signal reflector or signal absorber.

31. The conjugate of claim 30 wherein the signal absorber is a contrast agent.

32. The conjugate of claim 31 wherein the contrast agent comprises a metal ion selected from Gd, Mn and Fe.

33. The conjugate of claim 26 wherein the dense star polymer is a polyamidoamine.

34. The conjugate of claim 33 wherein the polyamidoamine is ester terminated.

35. The conjugate of claim 26 wherein the dense star polymer is a polyethyleneimine.

36. The conjugate of claim 35 wherein the polyethyleneimine has a methylene carboxylate surface.

37. The conjugate of claim 35 wherein the polyethyleneimine has an acetate surface.

38. The conjugate of claim 35 wherein the polyethyleneimine has a surface of a polyamidoamine.

39. The conjugate of claim 26 wherein the dendrimer has its surface modified with a functional group.

40. A formulation which comprises a conjugate of claim 26 having at least one pharmaceutically acceptable diluent or carrier present.

41. The formulation of claim 40 in which y is at least 2, and at least two units of carried material M are different carried materials.

42. A formulation which comprises a conjugate of claim 26 having at least one agriculturally acceptable diluent or carrier present.

43. The formulation of claim 42 in which y is at least 2, and at least two units of carried material M are different carried materials.

56. A process for preparing a conjugate as defined in claim 26 which comprises the reaction of P, having reactive moieties, with an aniline moiety, which may have the NH₂ group protected by an N-phthalimide of the formula ##STR21##

71. A process for preparing a conjugate as defined in claim 26 which comprises the reaction of P, having reactive moieties, which may have the NH₂ group protected by any protecting group used for amines which is inert under the conditions used for dense star polymer synthesis.

114. The conjugate of claim 26 wherein the dendrimer is solvent soluble.

44. A process for preparing

View Dependent Claims (45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55)

45. The process of claim 44 wherein the temperature is from room temperature to reflux.

46. The process of claim 44 wherein the suitable solvent is water, methanol, ethanol, chloroform, acetonitrile, toluene, dimethylsulfoxide, N-methylpyrrolidinone or dimethylformamide.

47. The process of claim 44 wherein the starburst polymer is a polyamidoamine.

48. The process of claim 47 wherein the polyamidoamine is ester terminated.

49. The process of claim 47 wherein the starburst polymer is a polyethyleneimine.

50. The process of claim 49 wherein the polyethyleneimine has a methylene carboxylate surface.

51. The process of claim 49 wherein the polyethyleneimine has an acetate surface.

52. The process of claim 49 wherein the polyethyleneimine has a surface of a polyamidoamine.

53. The process of claim 44 wherein the starburst polymer is a polyether.

54. The process of claim 44 wherein the starburst polymer has been further reacted to have its surface modified with a functional group.

55. The process of claim 44 wherein at least one of the carried materials is associated with the dendrimer by the reaction of at least one amine on the dendrimer with an α

,β

-unsaturated ester, α

,β

-unsaturated ketone, α

,β

-unsaturated nitrile, α

,β

-unsaturated carboxyl, isothiocyanato, or epoxide moiety.

57. A process for preparing a polyethyleneimine which comprises reacting a polyethyleneiminemethane sulfonamide with hydrochloric acid.

58. A process for purifying a dendrimer of at least generation 3.0 having a solvent present which comprises removing the solvent by ultrafiltration using a membrane.

59. The process of claim 58 wherein the solvent is ethylenediamine.

63. A method for preparing an anhydrous dendrimer polyethylenimine of at least generation 3.0 which comprises the steps of:

(a) acid cleavage of dendrimer polyethylenimine methane sulfonamides;

(b) recovery of a product from step (a) by removal of the solvent and excess acid by distillation;

(c) neutralizing the product of step (b);

(d) dissolving the neutralized product obtained in step (c) by adding a solvent selected from the group toluene, benzene, xylene or mesitylene; and

(e) removal of the solvent by azeotropic distillation.

64. The method of claim 63 wherein the acid in step (a) is hydrochloric, trifluoroacetic, fluorosulfonic, or chlorosulfonic acid.

65. The method of claim 63 wherein the acid is hydrochloric acid.

66. The method of claim 63 wherein the solvent in step (d) is toluene.

67. A method for preparing an anhydrous dendrimer polyethylenimine of at least generation 3.0 which comprises dissolving the dendrimer polyethylenimine in a solvent selected from the group toluene, benzene, xylene or mesitylene and removing the solvent by azeotropic distillation.

68. A solvent soluble dense star polymer conjugate which comprises at least one dense star polymer of at least generation 3.0 and at least one carried material, other than a dye or dye moiety, which is attached to or linked to the surface of the dense star polymer or encapsulated within the interior of the dense star polymer by means of covalent bonding, hydrogen bonding, adsorption, absorption, metallic bonding, van der Waals forces, ionic bonding, coulombic forces, hydrophobic or hydrophilic forces, or any combination thereof,

with the proviso that the carried material maintains its effectiveness in the conjugate.

69. The conjugate of claim 68 wherein the dense star polymer is radially symmetrical.

70. The conjugate of claim 68 wherein the dense star polymer has at least one core branch emanating from a core, said branch having at least one terminal group provided that

(1) the ratio of terminal groups to the core branches is two or greater,

(2) the density of terminal groups per unit volume in the polymer is at least 1.5 times that of an extended conventional star polymer having similar core and monomeric moieties and a comparable molecular weight and number of core branches, each of such branches of the extended conventional star polymer bearing only one terminal group, and

(3) the dense star polymer has a molecular volume that is no more than about 80 percent of the molecular volume of said extended conventional star polymer as determined by dimensional studies using scaled Corey-Pauling molecular models, and has regular dendritic branching.

72. A dense star polymer conjugate which comprises at least one dense star polymer of at least generation 3.0 associated with at least one unit of at least one carried material selected from the group consisting of a signal generator, a signal reflector, a signal absorber, a fragrance, and chelator, with the proviso that the carried material maintains its effectiveness as a signal generator, signal reflector, signal absorber, fragrance or chelator, in the conjugate.

73. The conjugate of claim 72 wherein the dense star polymer is a dendrimer.

74. The conjugate of claim 73 wherein the dendrimer is radially symmetrical.

75. The conjugate of claim 73 for use as a diagnostic agent.

76. The conjugate of claim 73 wherein the dendrimer is solvent soluble.

77. A dense star polymer conjugate of the formula:

space="preserve" listing-type="equation">(P).sub.x *(M).sub.y (I)

wherein each P represents a dendrimer of at least generation 3.0;

x represents an integer of 1 or greater;

each M represents a unit of a carried material selected from the group consisting of a signal generator, a signal reflector, a signal absorber, a fragrance, and chelator, said carried material can be the same carried material or a different carried material, with the proviso that the carried material maintains its effectiveness as a signal generator, signal reflector, signal absorber, fragrance or chelator, in the conjugate;

y represents an integer of 1 or greater; and

* indicates that the carried material is associated with the dendrimer.

78. The conjugate of claim 77 wherein the dendrimer is radially symmetrical.

79. The conjugate of claim 77 wherein the dendrimer is solvent soluble.

80. The conjugate of claim 77 wherein M is a signal generator.

81. The conjugate of claim 80 wherein M is a signal generator selected from the group consisting of electromagnetic radiation emitters, phosphorescent entities, fluorescent entities, gamma emitters, neutron emitters, positron emitters, β

-particle emitters, and α

-particle emitters.

82. The conjugate of claim 81 wherein M is an electromagnetic radiation emitter selected from the group X-ray emitters, ultraviolet radiation emitters, and infrared radiation emitters.

83. The conjugate of claim 81 wherein M is a gamma emitter, neutron emitter, positron emitter, β

-particle emitter or α

-particle emitter selected from the group consisting of a radionuclide, a chelated radionuclide and boron.

84. The conjugate of claim 77 wherein M is a signal reflector.

85. The conjugate of claim 84 wherein M is a signal reflector selected from the group consisting of paramagnetic entities, magnetic entities, chelated metals, chelated paramagnetic entities, chelated magnetic entities, nitroxyl radicals, and NMR shift reagents.

86. The conjugate of claim 77 wherein M is a signal absorber.

87. The conjugate of claim 86 wherein M is a signal absorber selected from the group consisting of contrast agents, neutron absorber, electron beam opacifiers, diagnostic opacifiers, aromatic UV absorber, boron, neutron absorber, phosphorescent entities, and fluorescent entities.

88. The conjugate of claim 77 wherein M is a fragrance.

89. The conjugate of claim 88 wherein M is a fragrance selected from the group consisting of perfumes and flavors.

90. The conjugate of claim 77 wherein M is a chelator.

91. The conjugate of claim 77 wherein P is a bridged dendrimer.

92. The conjugate of claim 77 wherein P is radially symmetrical.

93. The conjugate of claim 77 for use as a diagnostic agent.

94. The conjugate of claim 77 for use as a reagent in positron emission tomography, computer aided tomography, or magnetic resonance imaging.

95. The conjugate of claim 84 wherein the signal reflector is a paramagnetic entity or chelated paramagnetic entity which is prepared by reaction of an α

,β

-unsaturated ester, an α

,β

-unsaturated ketone, an α

,β

-unsaturated nitrile, an α

,β

-unsaturated carboxyl, isothiocyanato or epoxide moiety.

96. The conjugate of claim 89 wherein the carried material is hydrophobic.

97. The conjugate of claim 89 wherein the carried material is hydrophilic.

98. The conjugate of claim 77 wherein x=1 and y=2 or more.

99. The conjugate of claim 77 wherein the molar ratio of any ionic M to P is 0.1-1,000:

1.

100. The conjugate of claim 84 wherein the signal reflector is a paramagnetic entity or chelated paramagnetic entity wherein the entity contains a metal ion selected from Gd, Mn and Fe.

101. The conjugate of claim 77 wherein P is a polyamidoamine.

102. The conjugate of claim 101 wherein the polyamidoamine is ester terminated.

103. The conjugate of claim 77 wherein P is a polyethyleneimine.

104. The conjugate of claim 103 wherein the polyethyleneimine has a methylene carboxylate surface.

105. The conjugate of claim 103 wherein the polyethyleneimine has an acetate surface.

106. The conjugate of claim 103 wherein the polyethyleneimine has a surface of a polyamidoamine.

107. The conjugate of claim 77 wherein P has its surface modified with a functional group.

108. A formulation which comprises a conjugate of claim 77 having at least one pharmaceutically acceptable diluent or carrier present.

109. The formulation of claim 108 in which y is at least 2, and at least two units of carried material M are different carried materials.

110. A formulation which comprises a conjugate of claim 77 having at least one agriculturally acceptable diluent or carrier present.

111. The formulation of claim 110 in which y is at least 2, and at least two units of carried material M are different carried materials.

112. A process for preparing