Nanoparticles complexed with functionalizable enhanced affinity ligands and use thereof

Abstract

Disclosed are functionalizable ligands, nanoparticles, preferably nanocrystals, complexed with ligands and their use for bio-imaging. A nano material includes a nanoparticle and at least one copolymer ligand. A ligand which is a copolymer of general formula (I): H-P[(A)x-co-(B)y]n-L-R.

1 . A nanomaterial comprising:
 a 2D nanocrystal; and   at least one ligand which is a copolymer of formula (I):
   H-Poly[(A) x -co-(B) y ] n -L-R 
 wherein 
 H represents a hydrogen atom; 
 co means copolymer; 
 Poly[(A) x -co-(B) y ]n designates a copolymer with a total of n monomers, a percentage x of said n monomers is monomer A and a percentage y of said n monomers is monomer B, wherein 
 monomer A represents an anchoring monomer having a side-chain comprising a first moiety M A  having affinity for the surface of a nanocrystal; 
 monomer B represents a hydrophilic monomer having a side-chain comprising a second moiety M B  being hydrophilic; 
 n represents a positive integer; 
 x and y are different from 0% of n and different from 100% of n; wherein x+y is equal to 100% of n; 
 R represents:
 a functional group selected from the group comprising —NH 2 , —COOH, —OH, —SH, —CHO, ketone, halide; activated ester such as for example N-hydroxysuccinimide ester, N-hydroxyglutarimide ester or maleimide ester; activated carboxylic acid such as for example acid anhydride or acid halide; isothiocyanate; isocyanate; alkyne; azide; glutaric anhydride, succinic anhydride, maleic anhydride; hydrazide; chloroformate, maleimide, alkene, silane, hydrazone, oxime and furan; or 
 a bioactive group selected from the group comprising avidin or streptavidin; antibody such as a monoclonal antibody or a single chain antibody; sugars; a protein or peptide sequence having a specific binding affinity for an affinity target, such as for example an avimer or an affibody (the affinity target may be for example a protein, a nucleic acid, a peptide, a metabolite or a small molecule), antigens, steroids, vitamins, drugs, haptens, metabolites, toxins, environmental pollutants, amino acids, peptides, proteins, aptamers, nucleic acids, nucleotides, peptide nucleic acid (PNA), folates, carbohydrates, lipids, phospholipid, lipoprotein, lipopolysaccharide, liposome, hormone, polysaccharide, polymers, polyhistidine tags, fluorophores; and 
 
 L represents a bound or a spacer selected from the group comprising alkylene, alkenylene, arylene or arylalkyl linking groups having 1 to 50 chain atoms, wherein the linking group can be optionally interrupted or terminated by —O—, —S—, —NR 7 —, wherein R 7  is H or alkyl, —CO—, —NHCO—, —CONH— or a combination thereof; or a spacer selected from the group comprising DNA, RNA, peptide nucleic acid (PNA), polysaccharide, peptide. 
   
     
     
         2 . The nanomaterial according to  claim 1 , wherein R represents an antibody. 
     
     
         3 . The nanomaterial according to  claim 1 , wherein said 2D nanocrystal is a nanoplatelet or a nanoplate. 
     
     
         4 . The nanomaterial according to  claim 1 , wherein said 2D nanocrystal is a core/shell semiconductor nanocrystal. 
     
     
         5 . The nanomaterial according to  claim 4 , wherein the core and the shell are selected from CdS, CdSe, CdTe, ZnS, ZnSe, ZnTe, HgS, HgSe, HgTe, PbS, PbSe, PbTe, CuInS 2 , CuInSe 2 , AgInS 2 , AgInSe 2 , CuS, Cu 2 S, Ag 2 S, Ag 2 Se, Ag 2 Te, InN, InP, InAs, InSb, In 2 S 3 , Cd 3 P 2 , Zn 3 P 2 , Cd 3 As 2 , Zn 3 As 2 , ZnO, AlN, AlP, AlAs, AlSb, GaN, GaP, GaAs, GaSb, FeS 2 , TiO 2 , Bi 2 S 3 , Bi 2 Se 3 , Bi 2 Te 3 , MoS 2 , WS 2 , VO 2 , and alloys and mixtures thereof. 
     
     
         6 . The nanomaterial according to  claim 5 , wherein the core and the shell are selected from CdS, CdSe, CdTe, ZnS, ZnSe, ZnTe, HgS, HgSe, HgTe, PbS, PbSe, PbTe, and alloys and mixtures thereof. 
     
     
         7 . The nanomaterial according to  claim 1 , wherein the ligand is of formula (II): 
       
         
           
           
               
               
           
         
         wherein 
         n, x, y, L and R are as defined in  claim 1 ; 
         R A  represents a group comprising the first moiety M A  having affinity for the surface of a nanocrystal; 
         R B  represents a group comprising the second moiety M B  being hydrophilic; 
         R 1 , R 2 , R 3 , R 4 , R 5  and R 6  represent each independently H or a group selected from the alkyl, alkenyl, aryl, hydroxyl, halogen, alkoxy and carboxylate, amide. 
       
     
     
         8 . The nanomaterial according to  claim 1 , wherein the ligand is of formula (I-e′): 
       
         
           
           
               
               
           
         
         wherein n, x, y, L and R are as defined in  claim 1 . 
       
     
     
         9 . The nanomaterial according to  claim 1 , wherein the ligand is of formula (I-f′): 
       
         
           
           
               
               
           
         
         wherein n, x, y and L are as defined in  claim 1 . 
       
     
     
         10 . A water-soluble composition comprising nanomaterials comprising:
 a 2D nanocrystal; and   at least one ligand which is a copolymer of formula (I):
   H-Poly[(A) x -co-(B) y ] n -L-R 
 wherein 
 H represents a hydrogen atom; 
 co means copolymer; 
 Poly[(A) x -co-(B) y ]n designates a copolymer with a total of n monomers, a percentage x of said n monomers is monomer A and a percentage y of said n monomers is monomer B, wherein 
 monomer A represents an anchoring monomer having a side-chain comprising a first moiety M A  having affinity for the surface of a nanocrystal; 
 monomer B represents a hydrophilic monomer having a side-chain comprising a second moiety M B  being hydrophilic; 
 n represents a positive integer; 
 x and y are different from 0% of n and different from 100% of n; wherein x+y is equal to 100% of n; 
 R represents:
 a functional group selected from the group comprising —NH 2 , —COOH, —OH, —SH, —CHO, ketone, halide; activated ester such as for example N-hydroxysuccinimide ester, N-hydroxyglutarimide ester or maleimide ester; activated carboxylic acid such as for example acid anhydride or acid halide; isothiocyanate; isocyanate; alkyne; azide; glutaric anhydride, succinic anhydride, maleic anhydride; hydrazide; chloroformate, maleimide, alkene, silane, hydrazone, oxime and furan; or 
 a bioactive group selected from the group comprising avidin or streptavidin; antibody such as a monoclonal antibody or a single chain antibody; sugars; a protein or peptide sequence having a specific binding affinity for an affinity target, such as for example an avimer or an affibody (the affinity target may be for example a protein, a nucleic acid, a peptide, a metabolite or a small molecule), antigens, steroids, vitamins, drugs, haptens, metabolites, toxins, environmental pollutants, amino acids, peptides, proteins, aptamers, nucleic acids, nucleotides, peptide nucleic acid (PNA), folates, carbohydrates, lipids, phospholipid, lipoprotein, lipopolysaccharide, liposome, hormone, polysaccharide, polymers, polyhistidine tags, fluorophores; and 
 
 L represents a bound or a spacer selected from the group comprising alkylene, alkenylene, arylene or arylalkyl linking groups having 1 to 50 chain atoms, wherein the linking group can be optionally interrupted or terminated by —O—, —S—, —NR 7 —, wherein R 7  is H or alkyl, —CO—, —NHCO—, —CONH— or a combination thereof; or a spacer selected from the group comprising DNA, RNA, peptide nucleic acid (PNA), polysaccharide, peptide. 
   
     
     
         11 . A method for bioimaging, biotargeting, medical imaging or biosensing comprising applying an effective amount of a nanomaterial according to  claim 1 . 
     
     
         12 . A method for manufacturing the nanomaterial according to  claim 1  comprising:
 a first step of complexation of 2D nanocrystals with an intermediate ligand being a weakly binding ligand or a small molecule ensuring the homogeneous dispersion of the nanocrystal into a solvent miscible in part with water; 
 a step of monophasic exchange at about 40° C. to about 100° C. in an aqueous medium overnight to remove the weak intermediate ligand and replace it by the ligand which is a copolymer of general formula (I):
   H-Poly[(A) x -co-(B) y ] n -L-R 
 wherein 
 H represents a hydrogen atom; 
 co means copolymer; 
 Poly[(A) x -co-(B) y ]n designates a copolymer with a total of n monomers, a percentage x of said n monomers is monomer A and a percentage y of said n monomers is monomer B, wherein 
 monomer A represents an anchoring monomer having a side-chain comprising a first moiety M A  having affinity for the surface of a nanocrystal; 
 monomer B represents a hydrophilic monomer having a side-chain comprising a second moiety M B  being hydrophilic; 
 n represents a positive integer; 
 x and y are different from 0% of n and different from 100% of n; wherein x+y is equal to 100% of n; 
 R represents:
 a functional group selected from the group comprising —NH 2 , —COOH, —OH, —SH, —CHO, ketone, halide; activated ester such as for example N-hydroxysuccinimide ester, N-hydroxyglutarimide ester or maleimide ester; activated carboxylic acid such as for example acid anhydride or acid halide; isothiocyanate; isocyanate; alkyne; azide; glutaric anhydride, succinic anhydride, maleic anhydride; hydrazide; chloroformate, maleimide, alkene, silane, hydrazone, oxime and furan; or 
 a bioactive group selected from the group comprising avidin or streptavidin; antibody such as a monoclonal antibody or a single chain antibody; sugars; a protein or peptide sequence having a specific binding affinity for an affinity target, such as for example an avimer or an affibody (the affinity target may be for example a protein, a nucleic acid, a peptide, a metabolite or a small molecule), antigens, steroids, vitamins, drugs, haptens, metabolites, toxins, environmental pollutants, amino acids, peptides, proteins, aptamers, nucleic acids, nucleotides, peptide nucleic acid (PNA), folates, carbohydrates, lipids, phospholipid, lipoprotein, lipopolysaccharide, liposome hormone, polysaccharide, polymers, polyhistidine tags, fluorophores; and 
 
 L represents a bound or a spacer selected from the group comprising alkylene, alkenylene, arylene or arylalkyl linking groups having 1 to 50 chain atoms, wherein the linking group can be optionally interrupted or terminated by —O—, —S—, —NR 7 —, wherein R 7  is H or alkyl, —CO—, —NHCO—, —CONH— or a combination thereof; or a spacer selected from the group comprising DNA, RNA, peptide nucleic acid (PNA), polysaccharide, peptide.