Nanoplatforms for arginase, indoleamine 2,3-dioxygenase and tryptophan 2,3-dioxygenase detection by posttranslational modification

Patent metadata

Field	Value
Publication number	US-10376599-B2
Application number	US-201615559543-A
Country	US
Kind code	B2
Filing date	Mar 18, 2016
Priority date	Mar 19, 2015
Publication date	Aug 13, 2019
Grant date	Aug 13, 2019

Abstract

A nanoplatform assembly for detection of arginase, indoleamine 2,3-dioxygenase 1, and/or tryptophan 2,3-dioxygenase. The nanoplatform comprises an oligopeptide, which is used as a linker between two particles. More preferably, the linker is comprised of an oligopeptide containing a substrate for the target enzyme, where the substrate is chemically or physically modified by the target enzyme (but not cleaved). A central particle with a plurality of oligopeptide-tethered detectable particles and a plurality of directly attached detectable particles is described. Posttranslational modification of the oligopeptide leads to changes in the detectable signals from the first and/or second particles in the nanoplatform, which can be correlated to enzyme activity and concentration.

First claim

We claim:

1. A nanoplatform assembly for detecting the presence of an active enzyme in a sample, wherein said enzyme is arginase, said nanoplatform comprising:
a central carrier particle;
a first particle;
a second particle; and
an oligopeptide linkage connecting said central carrier particle and said second particle,
wherein said first particle is directly attached to said central carrier particle, and wherein said oligopeptide linkage comprises a substrate sequence for said enzyme, wherein said substrate sequence comprises a sequence according to (I) or (II):
(X) n (R) m (X) l (SEQ ID NO:1),  (I)

where:
each R: arginine;
each X: any amino acid;
m: 3-10;
n: 1-6; and
l: 1-6; or
(X) n (RJ) m (X) l (SEQ ID NO:2),  (II)

where:
each R: arginine;
each X: any amino acid;
each J: E, A, L, M, K, R, Q, or H;
m: 2-20;
n: 1-6; and
l: 1-6.

2. A nanoplatform assembly for detecting the presence of an active enzyme in a sample, wherein said enzyme is indoleamine 2,3-dioxygenase 1, said nanoplatform comprising:
a central carrier particle;
a first particle;
a second particle; and
an oligopeptide linkage connecting said central carrier particle and said second particle,
wherein said first particle is directly attached to said central carrier particle, and wherein said oligopeptide linkage comprises a substrate sequence for said enzyme, said substrate sequence comprising a sequence according to (III) or (IV):
(X) n (W) m (X) l (SEQ ID NO:3),  (III)

where:
each W: L- or D-tryptophan;
each X: any amino acid;
m: 3-10
n: 1-6; and
l: 1-6; or
(X) n (WU) m (X) l (SEQ ID NO:4),  (IV)

where:
each W: L- or D-tryptophan;
each X: any amino acid;
each U: G, N, D, P, or S;
m: 2-20;
n: 1-6; and
l: 1-6.

3. A nanoplatform assembly for detecting the presence of an active enzyme in a sample, wherein said enzyme is tryptophan 2,3-dioxygenase, said nanoplatform comprising:
a central carrier particle;
a first particle;
a second particle; and
an oligopeptide linkage connecting said central carrier particle and said second particle,
wherein said first particle is directly attached to said central carrier particle, and wherein said oligopeptide linkage comprises a substrate sequence for said enzyme, said substrate sequence comprises a sequence according to (III) and/or (IV):
(X) n (W) m (X) l (SEQ ID NO:3),  (III)

where:
each W: L-tryptophan;
each X: any amino acid;
m: 3-10
n: 1-6; and
l: 1-6; or
(X) n (WU) m (X) l (SEQ ID NO:4),  (IV)

where:
each W: L-tryptophan;
each X: any amino acid;
each U: G, N, D, P, or S;
m: 2-20;
n: 1-6; and
l: 1-6.

4. The nanoplatform assembly of claim 1 , comprising a plurality of said first particle and a plurality of said second particle, each of said first particles being directly attached to said central carrier particles, and each of said second particles being connected to said central carrier particle by respective oligopeptide linkages.

5. The nanoplatform assembly of claim 1 , wherein said first and second particles are each individually selected from the group consisting of nanoparticles, chromophores/luminophores, quantum dots, viologens, and combinations thereof.

6. The nanoplatform assembly of claim 1 , wherein said central carrier particle is a metal or non-metal nanoparticle.

7. The nanoplatform assembly of claim 6 , wherein said nanoparticle is a core/shell nanoparticle comprising a metal or metal-alloy core and a metal shell.

8. The nanoplatform assembly of claim 7 , wherein said core is selected from the group consisting of Au, Ag, Cu, Co, Fe, and Pt, and said shell is selected from the group consisting of Au, Ag, Cu, Co, Fe, Pt, the metal oxides thereof, and combinations thereof.

9. The nanoplatform assembly of claim 6 , wherein said nanoparticle is a stabilized nanoparticle comprising an organic monolayer coating, said first particle being directly attached to said coating.

10. The nanoplatform assembly of claim 2 , comprising a plurality of said first particle and a plurality of said second particle, each of said first particles being directly attached to said central carrier particles, and each of said second particles being connected to said central carrier particle by respective oligopeptide linkages.

11. The nanoplatform assembly of claim 2 , wherein said first and second particles are each individually selected from the group consisting of nanoparticles, chromophores/luminophores, quantum dots, viologens, and combinations thereof.

12. The nanoplatform assembly of claim 2 , wherein said central carrier particle is a metal or non-metal nanoparticle.

13. The nanoplatform assembly of claim 12 , wherein said nanoparticle is a core/shell nanoparticle comprising a metal or metal-alloy core and a metal shell.

14. The nanoplatform assembly of claim 13 , wherein said core is selected from the group consisting of Au, Ag, Cu, Co, Fe, and Pt, and said shell is selected from the group consisting of Au, Ag, Cu, Co, Fe, Pt, the metal oxides thereof, and combinations thereof.

15. The nanoplatform assembly of claim 12 , wherein said nanoparticle is a stabilized nanoparticle comprising an organic monolayer coating, said first particle being directly attached to said coating.

16. The nanoplatform assembly of claim 3 , comprising a plurality of said first particle and a plurality of said second particle, each of said first particles being directly attached to said central carrier particles, and each of said second particles being connected to said central carrier particle by respective oligopeptide linkages.

17. The nanoplatform assembly of claim 3 , wherein said first and second particles are each individually selected from the group consisting of nanoparticles, chromophores/luminophores, quantum dots, viologens, and combinations thereof.

18. The nanoplatform assembly of claim 3 , wherein said central carrier particle is a metal or non-metal nanoparticle.

19. The nanoplatform assembly of claim 18 , wherein said nanoparticle is a core/shell nanoparticle comprising a metal or metal-alloy core and a metal shell.

20. The nanoplatform assembly of claim 19 , wherein said core is selected from the group consisting of Au, Ag, Cu, Co, Fe, and Pt, and said shell is selected from the group consisting of Au, Ag, Cu, Co, Fe, Pt, the metal oxides thereof, and combinations thereof.

21. The nanoplatform assembly of claim 18 , wherein said nanoparticle is a stabilized nanoparticle comprising an organic monolayer coating, said first particle being directly attached to said coating.

Assignees

• Univ Kansas State

Inventors

• Bossmann Stefan H
• Troyer Deryl L
• Malalasekera Aruni P
• Wang Hongwang
• Wendel Sebastian O
• Zhu Gaohong

Classifications

• A61K49/0032

  Methine dyes, e.g. cyanine dyes · CPC title

• A61K49/0093 Primary

  Nanoparticle, nanocapsule, nanobubble, nanosphere, nanobead, i.e. having a size or diameter smaller than 1 micrometer, e.g. polymeric nanoparticle · CPC title

• A61K49/0056

  Peptides, proteins, polyamino acids · CPC title

• C12Y305/03001

  Arginase (3.5.3.1) · CPC title

• G01N33/573

  for enzymes or isoenzymes · CPC title