Enzymatic conjugation of polypeptides

Patent metadata

Field	Value
Publication number	US-2018071402-A1
Application number	US-201715654585-A
Country	US
Kind code	A1
Filing date	Jul 19, 2017
Priority date	Dec 23, 2011
Publication date	Mar 15, 2018
Grant date	—

Abstract

The present application relates to methods for the functionalization of immunoglobulins, in particular with drugs. Also disclosed herein are linking reagents, functionalized antibodies, pharmaceutical compositions, and method of treating disease and/or conditions.

First claim

1 - 53 . (canceled)

54 . A composition comprising a population of antibodies or antibody fragments, wherein each member of the population has the same primary amino acid sequence, and wherein at least 90% of the antibodies or antibody fragments in the composition have (m) functionalized acceptor glutamine residues (Q) per antibody or fragment, wherein m is an integer selected from 2 or 4, and wherein each of the functionalized acceptor glutamine residues has the structure of Formula IVb,
(Q)-NH—(C)n-X-L-(V-(Y-(M)z)q)r   Formula IVb
or a pharmaceutically acceptable salt or solvate thereof, wherein: Q is a glutamine residue present within or substituted into the primary amino acid sequence of a constant region of the antibodies or antibody fragments; (C)n is a substituted alkyl chain, an unsubstituted alkyl chain, a substituted heteroalkyl chain, or an unsubstituted heteroalkyl chain; n is an integer selected from among the range of 2 to 20; X is NH, O, S, absent, or a bond; L is independently absent, a bond or a continuation of a bond, or a carbon comprising framework of 1 to 200 atoms substituted at one or more atoms; r is an integer selected from among 1, 2, 3 or 4; q is an integer selected from among 1, 2, 3 or 4; z is an integer selected from among 1, 2, 3 or 4; V is independently absent, a bond or a continuation of a bond, a non-cleavable moiety or a conditionally-cleavable moiety; Y is independently absent, a bond or a continuation of a bond, or a spacer system which is comprised of 1 or more spacers; M is (RR′)-L′-(V′-(Y′-(Z)z′)q′)r′, wherein R is a reactive moiety; (RR′) is an addition product between R and a complementary reactive moiety R′; L′ is independently a carbon comprising framework of 1 to 200 atoms substituted at one or more atoms; V′ is independently absent, a bond or a continuation of a bond, a non-cleavable moiety or a conditionally-cleavable moiety; Y′ is independently absent, a bond or a continuation of a bond, or a spacer system which is comprised of 1 or more spacers; Z is a compound having a molecular weight of at least 300 g/mol and comprising a polycyclic group, tricycle or one or more macrocycles; and z′, q′ and r′ are each independently an integer selected from among 1, 2, 3 or 4, wherein the antibodies or antibody fragments specifically bind to an antigen present on immune cells that are contributing to inflammatory or autoimmune disease.

55 . The composition of claim 54 , wherein RR′ is a thio-maleimide (or halo-acetamide) addition product, a Staudinger ligation product, a Huisgen 1,3-cycloaddition product, a Diels-Alder cycloaddition adduct, or any high yield selective amidation or imidization reaction product.

56 . The composition of claim 54 , wherein said acceptor glutamine residue is flanked at the +2 position by a non-aspartic acid residue.

57 . The composition of claim 54 , wherein L comprises a (CH 2 —CH 2 —O—)x group, wherein x is an integer from among the range of 1 to 24.

58 . The composition of claim 54 , wherein the groups —(C)n-X-L-collectively comprise a structure (CH 2 —CH 2 —O—)x, wherein x is an integer from among the range of 3 to 24.

59 . The composition of claim 54 , wherein Z is an anti-inflammatory agent.

60 . The composition of claim 54 , wherein m is 4.

61 . A composition comprising a population of antibodies or antibody fragments each comprising at least one acceptor glutamine on each heavy chain, wherein at least 90% of the antibodies or antibody fragments in the composition comprise on each heavy chain two functionalized acceptor glutamine residues (Q) having the structure of Formula IVb of claim 25 and wherein each member of the population has the same primary amino acid sequence.

62 . A composition comprising a population of antibodies or antibody fragments, wherein at least 90% of the antibodies or antibody fragments in the composition comprise on each heavy chain at least one functionalized acceptor glutamine residues having the structure of Formula IVb,
(Q)-NH—(C)n-X-L-(V-(Y-(M)z)q)r   Formula IVb
or a pharmaceutically acceptable salt or solvate thereof, wherein: Q is a glutamine residue present within or substituted into the primary amino acid sequence of a constant region of the antibodies or antibody fragments; (C)n is a substituted alkyl chain, an unsubstituted alkyl chain, a substituted heteroalkyl chain, or an unsubstituted heteroalkyl chain; n is an integer selected from among the range of 2 to 20; X is NH, O, S, absent, or a bond; L is independently absent, a bond or a continuation of a bond, or a carbon comprising framework of 1 to 200 atoms substituted at one or more atoms; r is an integer selected from among 1, 2, 3 or 4; q is an integer selected from among 1, 2, 3 or 4; z is an integer selected from among 1, 2, 3 or 4; V is independently absent, a bond or a continuation of a bond, a non-cleavable moiety or a conditionally-cleavable moiety; Y is independently absent, a bond or a continuation of a bond, or a spacer system which is comprised of 1 or more spacers; M is (RR′)-L′-(V′-(Y′-(Z)z′)q′)r′, wherein R is a reactive moiety; (RR′) is an addition product between R and a complementary reactive moiety R′; L′ is independently a carbon comprising framework of 1 to 200 atoms substituted at one or more atoms; V′ is independently absent, a bond or a continuation of a bond, a non-cleavable moiety or a conditionally-cleavable moiety; Y′ is independently absent, a bond or a continuation of a bond, or a spacer system which is comprised of 1 or more spacers; Z is a compound having a molecular weight of at least 300 g/mol and comprising a polycyclic group, tricycle or one or more macrocycles; and z′, q′ and r′ are each independently an integer selected from among 1, 2, 3 or 4wherein at least 90% of the antibodies or antibody fragments in the composition have (m) functionalized acceptor glutamine residues (Q) per antibody or fragment, wherein m is an integer selected from 2 or 4, wherein the antibodies or antibody fragments specifically bind to an antigen present on immune cells that are contributing to inflammatory or autoimmune disease, and wherein each member of the population has the same primary amino acid sequence.

63 . A pharmaceutical formulation comprising the composition of claim 54 , and a pharmaceutically acceptable carrier.

64 . A method of treating a disease comprising administering to a mammal a composition of claim 63 .

65 . The composition of claim 54 , wherein Z is a pyrrolobenzodiazepine.

66 . The composition of claim 54 , wherein R is a reagent capable of undergoing a Huisgen 1,3-cycloaddition.

67 . The composition of claim 66 , wherein RR′ is 1,4-disubstituted-1,2,3-triazole.

68 . The composition of claim 55 , wherein the thio-maleimide addition product is N,S-disubstituted-3-thio-pyrrolidine-2,5-dione.

69 . The composition of claim 55 , wherein the Staudinger ligation product is N,3-substituted-5-dipenylphosphinoxide-benzoic amide or N,4-substituted-5-dipenylphosphinoxide-benzoic amide.

70 . The composition of claim 55 , wherein the Huisgen 1,3-cycloaddition product is selected from the group consisting of N,S-disubstituted-3-thio-pyrrolidine-2,5-dione, 1,4-disubstituted-1,2,3-triazole, 3,5-disubstituted-isooxazole, and 3,5-disubstituted-tetrazol.

71 . The composition of claim 55 , wherein the Diels-Alder cycloaddition adduct is the 2,4-cycloaddition product between a 9-substituted anthracene or 3-substituted 1,2,4,5-tetrazine with a compound selected from the group

Assignees

• Innate Pharma
• Scherrer Inst Paul

Inventors

• Bregeon Delphine
• Dennler Patrick
• Belmant Christian
• Fischer Eliane
• Gauthier Laurent
• Romagné Francois
• Schibli Roger

Classifications

• A61P35/00

  Antineoplastic agents · CPC title

• C07C217/08

  the oxygen atom of the etherified hydroxy group being further bound to an acyclic carbon atom · CPC title

• A61K47/6817

  Toxins · CPC title

• A61K47/6813

  the drug being a peptidic cytokine, e.g. an interleukin or interferon · CPC title

• C07C233/62

  having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by amino groups · CPC title