Loaded latex optical molecular imaging probes containing lipophilic large stokes shift dyes

• US 8,906,354 B2
• Filed: 03/08/2011
• Issued: 12/09/2014
• Est. Priority Date: 02/28/2007
• Status: Active Grant

• Current Assignee: Bruker Biospin Corporation (Bruker Corporation)
• Original Assignee: Bruker Biospin Corporation (Bruker Corporation)
• Inventors: Ji, Tao, Schmitthenner, Hans F., Yang, Yonghong, Harder, John W., Leon, Jeffrey W., Harrison, William J., Kelley, Brian J., Bennett, James R., Stegman, David A., Wang, Ruizheng
• Primary Examiner(s): DICKINSON, PAUL W
• Application Number:
  US13/043,057
• Publication Number:
  US 20120058050A1
• Time in Patent Office: 1,372 Days
• Field of Search: None
• US Class Current: 424/78.17
• CPC Class Codes: A61K 49/0032 Methine dyes, e.g. cyanine dyes
• CPC Class Codes: A61K 49/0054 Macromolecular compounds, i.e. oligomers, polymers, dendrimers
• CPC Class Codes: A61K 49/0093 Nanoparticle, nanocapsule, nanobubble, nanosphere, nanobead, i.e. having a size or diameter smaller than 1 micrometer, e.g. polymeric nanoparticle
• CPC Class Codes: B82Y 15/00 Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors

Abstract

The present invention relates to a loaded particle comprising at least one fluorescent dye, and in particular, a fluorescent dye with a large Stokes shift. The invention further relates to a method for producing an loaded latex particle, loaded with a fluorescent dye having a large stokes shift. In addition, the present invention relates to latex particles loaded with fluorescent dyes that are organic solvent soluble and insoluble in water. In a preferred embodiment, when the dyes are loaded into the water soluble latex particle, an increase is observed in quantum yield of fluorescence as compared to the quantum yield of the dye in aqueous solvent.

Claims

1. A nanolatex particle comprising a latex material made from a mixture represented by formula:

(X)m-(Y)n-(Z)o-(W)p

wherein Y is at least one monomer with at least two ethylenically unsaturated chemical functionalities;

Z is at least one polyethylene glycol macromonomer with an average molecular weight of between 300 and 10,000;

W is an ethylenic monomer different from X, Y, or Z; and

X is at least one water insoluble, alkoxethyl containing monomer represented by formula;

2. The nanolatex particle of claim 1, wherein the water dispersible, near-infrared tricarbocyanine, enamine-functionalized dye compound having a Stokes shift of greater than 50 nm is represented by the following general formula:

3. The nanolatex particle of claim 1, wherein the water dispersible, near-infrared tricarbocyanine, enamine-functionalized dye compound having a Stokes shift of greater than 50 nm is represented by the following general formula:

4. The nanolatex particle of claim 2, wherein the water dispersible, near-infrared tricarbocyanine, enamine-functionalized dye compound having a Stokes shift of greater than 50 nm is represented by the following formula:

5. The nanolatex particle of claim 2, wherein the water dispersible, near-infrared tricarbocyanine, enamine-functionalized dye compound having a Stokes shift of greater than 50 nm is represented by the following formula:

6. The nanolatex particle of claim 1, wherein said Z monomer is a polyethylene glycol backbone chain comprising at least one functional end group.

7. The nanolatex particle of claim 1, wherein Z is a polyethylene glycol macromonomer represented by a general formula:

8. The nanolatex particle of claim 1, wherein Z is a polyethylene glycol macromonomer represented by a general formula:

9. The nanolatex particle of claim 1, wherein said polyethylene glycol macromonomer is a polyethylene glycol derivative represented by formula:

10. The nanolatex particle of claim 1, further comprising a targeting agent.

11. The nanolatex particle of claim 1, further comprising a biomolecule.

12. The nanolatex particle of claim 1, further comprising a diagnostic agent.

13. The nanolatex particle of claim 1, further comprising a therapeutic agent.

14. The nanolatex particle of claim 1, wherein said nanolatex particle is biocompatible.

15. The nanolatex particle of claim 1, wherein said fluorescent dye compound is organic solvent soluble and insoluble in water.

16. The nanolatex particle of claim 1, wherein X is methoxyethyl methacrylate.

17. The nanolatex particle of claim 1, wherein m ranges between 45-60 percent by weight.

18. The nanolatex particle of claim 1, wherein;

n ranges between 2-6 percent by weight.

19. The nanolatex particle of claim 1, wherein o ranges between 40-50 percent by weight.

20. The nanolatex particle of claim 1, wherein Y is a monomer containing at least two ethylenically unsaturated chemical functionalities.

21. The nanolatex particle of claim 20, wherein said functionalities are selected from the group consisting of:

vinyl groups;

acrylates;

methacrylates;

acrylamides;

methacrylamides;

allyl groups;

vinyl ethers; and

vinyl esters.

22. The nanolatex particle of claim 20, wherein said Y monomer is selected from the group consisting of:

aromatic divinyl compounds, such as divinylbenzene, divinylnaphthalene, and derivatives thereof;

diethylene carboxylate esters and amides, such as ethylene glycol dimethacrylate, diethylene glycol diacrylate, 1,4 butanediol diacrylate, 1,4 butanediol dimethacrylate, 1,3 butylene glycol diacrylate, 1,3 butylene glycol dimethacrylate, cyclohexane dimethanol diacrylate, cyclohexane dimethanol dimethacrylate, diethylene glycol diacrylate, diethylene glycol dimethacrylate, dipropylene glycol diacrylate, dipropylene glycol dimethacrylate, ethylene glycol diacrylate, ethylene glycol dimethacrylate, 1,6 hexanediol diacrylate, 1,6 hexanediol dimethacrylate, neopentyl glycol diacrylate, neopentyl glycol dimethacrylate, tetraethylene glycol diacrylate, tetraethylene glycol dimethacrylate, triethylene glycol diacrylate, triethylene glycol dimethacrylate, tripropylene glycol diacrylate, tripropylene glycol dimethacrylate, pentaerythritol triacrylate, trimethylolpropane triacrylate, trimethylolpropane trimethacrylate, dipentaerythritol pentaacrylate, di-trimethylolpropane tetraacrylate, and pentaerythritol tetraacrylate;

divinyl esters, such as divinyl adipate;

divinyl compounds, such as divinyl sulfide, divinyl sulfone compounds of allyl methacrylate, allyl acrylate, cyclohexanedimethanol divinyl ether diallylphthalate, and diallyl maleate; and

dienes such as butadiene, isoprene, and mixtures thereof.

23. The nanolatex particle of claim 1, wherein said W monomer is a water-soluble monomer selected from the group consisting of:

2-phosphatoethyl acrylate potassium salt;

3-phosphatopropyl methacrylate ammonium salt;

vinylphosphonic acid, and salts thereof;

vinylcarbazole;

vinylimidazole;

vinylpyrrolidone;

vinylpyridines;

acrylamide;

methacrylamide;

maleic acid and salts thereof;

sulfopropyl acrylate and methacrylate;

acrylic acid and salts thereof;

methacrylic acid and salts thereof;

N-vinylpyrrolidone;

acrylic esters of alkylphosphonates, styrenics, acrylic, and methacrylic monomers containing amine or ammonium functionalities;

methacrylic esters of alkylphosphonates, styrenics, acrylic and methacrylic monomers containing amine or ammonium functionalities;

styrenesulfonic acid and salts thereof;

acrylic and methacrylic esters of alkylsulfonates, vinylsulfonic acid, and salts thereof;

vinylpyridines;

hydroxyethyl acrylate;

glycerol acrylate;

methacrylate esters;

(meth)acrylamide; and

N-vinylpyrrolidone.

24. The nanolatex particle of claim 1, wherein said W monomer is a water-insoluble monomer selected from the group consisting of methyl methacrylate;

ethyl methacrylate;

isobutyl methacrylate;

2-ethylhexyl methacrylate;

benzyl methacrylate;

cyclohexyl methacrylate;

glycidyl methacrylate;

acrylic/acrylate esters such as methyl acrylate, ethyl acrylate, isobutyl acrylate, 2-ethylhexyl acrylate, benzyl methacrylate, phenoxyethyl acrylate, cyclohexyl acrylate, and glycidyl acrylate;

styrenics such as styrene, a-methylstyrene, ethylstyrene, 3- and 4-chloromethylstyrene, halogen-substituted styrenes, and alkyl-substituted styrenes;

vinyl halides;

vinylidene halides;

N-alkylated acrylamides;

methacrylamides;

vinyl esters, such as vinyl acetate and vinyl benzoate; and

allyl alcohol and its ethers and esters.

25. The nanolatex particle of claim 1, wherein said Z monomer is a polyethylene glycol macromonomer having a molecular weight of between 500 and 5000.

26. The nanolatex particle of claim 1, wherein said Z monomer is a polyethylene glycol backbone chain with specific functional end groups at each end, which allows said polyethylene glycol to act as a linking group between two materials through said two functional end groups.

27. The nanolatex particle of claim 1, wherein said Z polyethylene glycol macromonomer contains a radical polymerizeable group at one end.

28. The nanolatex particle of claim 27, wherein said radical polymerizeable group is selected from the group consisting of:

methacrylate;

acrylate;

acrylamide;

methacrylamide;

styrenic;

allyl;

vinyl;

maleimide; and

maleate ester.

29. The nanolatex particle of claim 26, wherein said Z polyethylene glycol macromonomer contains a reactive chemical functionality at one end servable as an attachment point for other chemical units.

30. The nanolatex particle of claim 29, wherein said reactive chemical functionality is selected from the group consisting of:

alcohols;

thiols;

carboxylic acids;

primary amines;

secondary amines;

vinylsulfonyls;

aldehydes;

epoxides;

hydrazides;

succinimidyl esters;

maleimides;

a-halo carbonyl moieties;

iodoacetyls;

isocyanates;

isothiocyanates; and

aziridines.

31. The nanolatex particle of claim 29, wherein said reactive chemical functionality is servable as an attachment point for linkers designed to form covalent bonds between the reactive chemical functionality on the loaded latex and an attachment group on a bio molecule.

32. The nanolatex particle of claim 31, wherein said attachment group on a bio molecule is selected from the group consisting of:

amine;

alcohol;

carboxylic acid from amino acids;

thiol from amino acids;

peptides;

protein;

cells;

RNA; and

DNA.

33. The nanolatex particle of claim 31, wherein said linkers are selected from the group consisting of:

hetero-bifunctional linkers and homo-bifunctional linkers.

34. The nanolatex particle of claim 31, wherein said linkers are selected from the group consisting of:

bis-sulfosuccinylsuberate, 3-[2-(aminoethyl)dithio]propionic acid;

p-azidobenzoylhydrazide;

bis-maleimidohexane;

N-succinimidyl-5-acetylthioacetate;

N-Sulfosuccinimidyl-4-azidophenyl-1-3′

-dithiopropionatte;

Succinimidyl 4-[p-maleimidophenyl]butyrate;

N-Succinimidyl[4-iodoacetyl]aminobenzoate;

Sulfosuccinimidyl-[perfluoroazidobenzamido]ethyl-1,3′

-dithiopropionate;

Succinimidyl 3-[bromoacetamido]propionate;

Sulfosuccinimidyl 2-[7-amino-4-methylcoumarin-3-acetamido]ethyl-1,3′

dithiopropionate;

3-(2-Pyridyldithio)propionyl hydrazide);

N-e-Maleimidocaproyloxy]succinimide ester;

N-[4-(p-Azidosalicylamido) butyl]-3′

-(2′

-pyridyldithio)propionamide;

Succinimidyl 4-[N-maleimidomethyl]cyclohexane-1-carboxylate;

4-(4-N-Maleimidophenyl)butyric acid hydrazide hydrochloride;

Disuccinimidyl suberate;

Lomant'"'"'s Reagent;

Sulfosuccinimidyl[2-6-(biotinamido)-2-(p-azidobenzamido)-hexanoamido]-ethyl-1,3′

-dithiopropionate;

B-[Tris(hydroxymethyl)phosphino]propionic acid (betaine);

(Sulfosuccinimidyl 4-[p-maleimidophenyl]butyrate);

Bis-Maleimidoethane;

Bis-[b-(4-Azidosalicylamido)ethyl]disulfide;

Succinimidyl 4-[N-maleimidomethyl]cyclohexane-1-carboxy-[6-amidocaproate];

N-[p-Maleimidophenyl]isocyanate;

m-Maleimidobenzoyl-N-hydroxysulfosuccinimide ester;

Sulfosuccinimidyl 4-N-maleimidomethyl cyclohexane-1-carboxylate;

Bis[sulfosuccinimidyl]suberate;

N-[g-Maleimidobutyryloxy]sulfosuccinimide ester;

N-succinimidyl 4-pentynoate; and

N-succinimidyl 4-azidoylbutanoate.

35. The nanolatex particle of claim 1, wherein said particle has a volume-average hydrodynamic diameter of less than 100 nm.

36. The nanolatex particle of claim 1, wherein said particle has a volume-average hydrodynamic diameter of from 8 to 50 nm.

37. The nanolatex particle of claim 1, wherein said fluorescent dye compound is present in an amount of from 0.01 to 5 percent by weight.

38. The nanolatex particle of claim 1, wherein said fluorescent dye emits in the range of from 600 to 900 nm.

39. The nanolatex particle of claim 1, wherein Y is divinyl benzene.

40. The nanolatex particle of claim 39, wherein W is ethylstyrene.

41. The nanolatex particle of claim 40, wherein X is methoxyethyl methacrylate.

42. The nanolatex particle of claim 41, wherein Z is poly(ethylene glycol) monomethyl ether methacrylate.

43. The nanolatex particle of claim 42, wherein the water dispersible, near-infrared tricarbocyanine, enamine-functionalized dye compound having a Stokes shift of greater than 50 nm is represented by the following formula:

44. The nanolatex particle of claim 42, wherein the water dispersible, near-infrared tricarbocyanine, enamine-functionalized dye compound having a Stokes shift of greater than 50 nm is represented by the following formula: