Nanoparticle immunoconjugates

Patent metadata

Field	Value
Publication number	US-2018133342-A1
Application number	US-201615564315-A
Country	US
Kind code	A1
Filing date	Apr 7, 2016
Priority date	Apr 7, 2015
Publication date	May 17, 2018
Grant date	—

Abstract

Disclosed herein are nanoparticle immunoconjugates useful for therapeutics and/or diagnostics. The immunoconjugates have diameter (e.g., average diameter) no greater than 20 nanometers (e.g., as measured by dynamic light scattering (DLS) in aqueous solution, e.g., saline solution). In certain embodiments, the conjugates are silica-based nanoparticles with single chain antibody fragments attached thereto.

First claim

1 . An immunoconjugate comprising:
a nanoparticle; and an antibody fragment conjugated to the nanoparticle, wherein the nanoparticle has a diameter no greater than 20 nanometers.

2 . The immunoconjugate of claim 1 , wherein the antibody fragment is covalently or non-covalently bonded to the nanoparticle via a linker or covalently or non-covalently bonded directly to the nanoparticle, or associated with the nanoparticle or a composition surrounding the nanoparticle, e.g., via van der Waals forces.

3 . The immunoconjugate of claim 1 , wherein the nanoparticle is coated with an organic polymer.

4 . The immunoconjugate of claim 1 , comprising a targeting peptide (e.g., alphaMSH, any peptide known to be immunomodulatory and anti-inflammatory in nature).

5 . The immunoconjugate of claim 1 , wherein the antibody fragment is in a range from about 5 kDa to about 25 kDa (e.g., from about 10 kDa to about 20 kDa, e.g., about 15 kDa) (e.g., wherein the antibody fragment comprises a functional single domain antibody fragment).

6 . The immunoconjugate of claim 1 , wherein the antibody fragment is from about 20 kDa to about 45 kDa (e.g., wherein the antibody fragment comprises a functional single chain antibody fragment).

7 . The immunoconjugate of claim 1 , wherein the antibody fragment is from about 40 kDa to about 80 kDa (e.g., from about 50 kDa to about 70 kDa, e.g., about 60 kDa) (e.g., wherein the antibody fragment comprises a functional fab fragment).

8 . The immunoconjugate of claim 1 , wherein the nanoparticle comprises silica.

9 . The immunoconjugate of claim 1 , wherein the nanoparticle comprises a silica-based core and a silica shell surrounding at least a portion of the core.

10 . The immunoconjugate of claim 1 , wherein the nanoparticle comprises a fluorescent compound within the core.

11 . The immunoconjugate of claim 1 , wherein the antibody fragment is a member selected from the set consisting of a recombinant antibody fragment (fAbs), a single chain variable fragment (scFv), and a single domain antibody (sdAb) fragment.

12 . The immunoconjugate of claim 11 , wherein the antibody fragment is a single chain variable fragment (scFv).

13 . The immunoconjugate of claim 11 , wherein the antibody fragment is a single domain (sdAb) fragment.

14 . The immunoconjugate of claim 1 , wherein the nanoparticle (a single nanoparticle) has from one to ten antibody fragments attached thereto.

15 . The immunoconjugate of claim 1 , wherein the antibody fragment is conjugated to the nanoparticle via a PEG moiety and a chelator.

16 . The immunoconjugate of claim 1 , wherein the nanoparticle has a diameter no greater than 15 nanometers.

17 . The immunoconjugate of claim 1 , wherein the nanoparticle has a diameter in a range from 1 nm to 20 nm.

18 . The immunoconjugate of claim 1 , wherein the antibody fragment comprises a member selected from the set consisting of anti-CEA scFv, anti-GPM/Ma, anti-VEGF-A, and anti-TNF-α (e.g., PEGylated).

19 . The immunoconjugate of claim 1 , wherein the immunoconjugate comprises one or more imaging agents.

20 . The immunoconjugate of claim 19 , wherein the one or more imaging agents comprise a PET tracer.

21 . The immunoconjugate of claim 19 , wherein the one or more imaging agents comprise a fluorophore.

22 . The immunoconjugate of claim 1 , further comprising a therapeutic agent.

23 . The immunoconjugate of claim 22 , wherein the therapeutic agent comprises a chemotherapy drug.

24 . The immunoconjugate of claim 22 , wherein the therapeutic agent comprises a radioisotope.

25 . The immunoconjugate of claim 24 , wherein the radioisotope is a member selected from the group consisting of 99m Tc, 111 In, 64 Cu, 67 Ga, 186 Re, 188 Re, 153 Sm, 177 Lu, 67 Cu, 123 I, 124 I, 125 I, 11 C, 1 3N, 15 O, 18 F, 186 Re, 188 Re, 153 Sm, 166 Ho, 177 Lu, 149 Pm, 90 Y, 213 Bi, 103 Pd, 109 Pd, 159 Gd, 140 La, 198 Au, 199 Au, 169 Yb, 175 Yb, 165 Dy, 166 Dy, 67 Cu, 105 Rh, 111 Ag, 89 Zr, 225 AC, and 192 Ir. .

26 . A method of treating a disease or condition, the method comprising administering to a subject a pharmaceutical composition comprising the immunoconjugate of claim 1 .

27 . The method of claim 26 , comprising
administering a therapeutic radioisotope (e.g., wherein the therapeutic radioisotope is attached to a second nanoparticle having a diameter no greater than 20 nanometers (e.g., as measured by dynamic light scattering (DLS) in aqueous solution, e.g., saline solution) (e.g., wherein the radioisotope is attached to the second nanoparticle via a second chelator)).

28 . A method of treating a disease or condition, the method comprising administering to a subject a pharmaceutical composition comprising the immunoconjugate of claim 24 (e.g., for combined radiation therapy and immunotherapy).

29 . The method of claim 28 , wherein the pharmaceutical composition further comprises a carrier.

30 . A method of in vivo imaging, the method comprising:
administering to a subject a composition comprising the immunoconjugate of c 1 aim 1 (e.g., such that the immunoconjugate preferentially collects in a particular region, e.g., near or within a particular tissue type, e.g., cancer), wherein the immunoconjugate comprises an imaging agent; and detecting the imaging agent.

31 . A method of making the immunoconjugate of claim 1 , the method comprising:
contacting a nanoparticle-PEG-thiol with a protein-maleimide, thereby producing the immunoconjugate.

32 . The method of claim 31 , further comprising:
reacting the nanoparticle with one or more compounds, the one or more compounds comprising a thiol moiety and an amine moiety (e.g., cysteine methyl ester or cysteamine-HCl), thereby producing a nanoparticle-PEG-amine; and reacting the nanoparticle-PEG-amine with SPDP, then removing a pyridine 2 -thione from the product (e.g., using TCEP), thereby producing the nanoparticle-PEG-thiol.

33 . A method of making the immunoconjugate of claim 1 , the method comprising:
modifying the antibody fragment (protein) with a first click reactive group (e.g., methyltetrazine-PEG4-NHS ester); modifying a nanoparticle-PEG-amine with a click partner of the first click reactive group (e.g., TCO-PEG4-NHS ester); and reacting the modified antibody fragment with the modified nanoparticle-PEG, thereby producing the immunoconjugate.

34 . The method of claim 33 , further comprising reacting the nanoparticle with one or more compounds, the one or more compounds comprising a thiol moiety and an amine moiety (e.g., cysteine methyl ester or cysteamine-HCl), thereby producing the nanoparticle-PEG-amine.

35 . A method of treating a disease or condition, the method comprising administering to a subject a composition comprising:
a nanoparticle; and a therapeutic radioisotope conjugated to the nanoparticle, wherein the nanoparticle has a diameter no greater than 20 nanometers.

36 . The method of claim 35 , comprising
administering immunotherapy, wherein the immunotherapy comprises administering to a subject a pharmaceutical composition comprising the immunoconjugate of claim 1 .

37 - 44 . (canceled)

Assignees

• Memorial Sloan Kettering Cancer Center
• Univ Cornell
• Univ Missouri

Inventors

• Yoo Barney
• Quinn Thomas P
• Bradbury Michelle S
• Wiesner Ulrich
• Lewis Jason
• Ma Kai
• Chen Feng

Classifications

• A61K51/10

  Antibodies or immunoglobulins; Fragments thereof {, the carrier being an antibody, an immunoglobulin or a fragment thereof, e.g. a camelised human single domain antibody or the Fc fragment of an antibody} · CPC title

• A61K51/0478 Primary

  complexes from non-cyclic ligands, e.g. EDTA, MAG3 · CPC title

• A61P35/00

  Antineoplastic agents · CPC title

• A61P37/02

  Immunomodulators · CPC title

• C07K16/40

  against enzymes · CPC title