Synaffix B.v.

Berry and Associates, Inc.

Sander Sebastiaan Van Berkel

Floris Louis Van Delft

Maria Antonia Wijdeven

Jorge Merijn Mathieu Verkade

Remon Van Geel

Petrus Josephus Jacobus Maria Van De Sande

Floris Petrus Johannes Theodorus Rutjes

David A Berry

Ryan Heesbeen

Jan Cornelis Maria Van Hest

Inge Catharina Josephina Hurkmans

Jorin Hoogenboom

Brian Maria Gerardus Janssen

Anna Agnieszka Wasiel

Remon Gvan Eel

Lianne Lelieveldt

Marjoke Froukje Debets

John Cooke Hodges

Jorge Verkade

The present invention concerns antibody-conjugates which are especially suitable for the targeting of PTK7-expressing cells, in particular tumour cells. The antibody-conjugates according to the invention have structure (1):-[()-{--]  1Herein, AB is an antibody capable of targeting PTK7-expressing tumours; L is a linker that links Z to D; Z is a connecting group; Lis -GlcNAc(Fuc)-(G)-S-(L)-, wherein G is a monosaccharide, j is an integer in the range of 0-10, S is a sugar or a sugar derivative, GlcNAc is N-acetylglucosamine and Fuc is fucose, w is 0 or 1, w' is 0, 1 or 2 and Lis —N(H)C(O)CH—, —N(H)C(O)CF— or —CH—; D is selected from the group consisting of anthracyclines, camptothecins, tubulysins, enediynes, amanitins, duocarmycins, maytansinoids, auristatins, eribulins, BCL-XL inhibitors, hemiasterlins, KSP inhibitors, TLR agonists, indolinobenzodiazepine dimers or pyrrolobenzodiazepine dimers (PBDs), and analogues or prodrugs thereof; b is 0 or 1; x is 1 or 2; and y is 1, 2, 3 or 4. The invention further concerns a method for preparing the antibody-conjugates of structure (1) and application of the antibody-conjugates of structure (1).

Antibody-conjugates for targeting of tumours expressing PTK7

The present invention concerns antibody-conjugate having general structure (2):AB-[(L)-{Z—(L)—(L)—(L)—(L)-D}]   (2)wherein AB is an antibody capable of targeting Trop-2-expressing tumours and D is selected from the group consisting of taxanes, anthracyclines, camptothecins, epothilones, mytomycins, combretastatins, vinca alkaloids, maytansinoids, enediynes such as calicheamicins, duocarmycins, tubulysins, amatoxins, bleomycins, dolastatins and auristatins, pyrrolobenzodiazepine dimers, indolinobenzodiazepine dimers, radioisotopes, therapeutic proteins and peptides (or fragments thereof), kinase inhibitors, MEK inhibitors, KSP inhibitors, and analogues or prodrugs thereof. These antibody-conjugates exhibit an improved therapeutic index. The invention further concerns a process for preparing the antibody-conjugate according to the invention, a method for targeting Trop-2-expressing cells, medical uses of the antibody-conjugates according to the invention.

Antibody-conjugates for targeting of tumours expressing Trop-2

The current invention concerns methods for the synthesis of 6-azido-6-deoxy-2-N-acetyl-monosaccharide-nucleoside diphosphate, in particular 6-azido-6-deoxy-2-N-acetyl-D-galactosamine-nucleoside diphosphate or 6-azido-6-deoxy-2-N-acetyl-D-glucosamine-nucleoside diphosphate. The synthesis method according to the invention is characterized by being highly efficient and high yielding. Also part of the present invention are key intermediates of this process.

Synthesis of 6-azido-6-deoxy-2-n-acetyl-hexosamine-nucleoside diphosphate

The present invention relates to a process for the enzymatic modification of a glycoprotein. The process comprises the step of contacting a glycoprotein comprising a glycan comprising a terminal GlcNAc-moiety, in the presence of glycosyltransferase that is, or is derived from, a β-(1,4)-N-acetylgalactosaminyltransferase, with a non-natural sugar-derivative nucleotide. The non-natural sugar-derivative nucleotide is according to formula (3): wherein A is selected from the group consisting of —N, —C(O)R, —(CH)C≡C—R, —SH, —SC(O)R, —SC(O)OR, —SC(S)OR, —F, —Cl, —Br —I, —OS(O)R, terminal C-Calkenyl groups, C-Ccycloalkenyl groups, C-Calkadienyl groups, terminal C-Callenyl groups and amino groups. The invention further relates to a glycoprotein obtainable by the process according to the invention, to a bioconjugate that can be obtained by conjugating the glycoprotein with a linker-conjugate, and to β-(1,4)-N-acetylgalactosaminyltransferases that can be used in preparing the glycoprotein according to the invention.

Process for the modification of a glycoprotein using a glycosyltransferase that is or is derived from a β(1,4)-n-acetylgalactosaminyltransferase

The invention relates to a novel linker for use in bioconjugates such as antibody-drug-conjugates. The linker according to the invention is represented by formula: wherein: The linker according to the invention is useful in the preparation of linker-conjugates and bioconjugates, and can be used for (a) improving conjugation efficiency in the preparation of the bioconjugate, (b) reducing aggregation during the preparation of the bioconjugate and/or of the bioconjugate, (c) increasing stability of the bioconjugate, and/or (d) increasing therapeutic index of the bioconjugate.

Sulfamide linkers for use in bioconjugates

The present invention is based on the surprising finding that the linker employed in a bioconjugate, such as an anti-body-drug conjugate, is not therapeutically inert but has an effect on the therapeutic index of the bioconjugate. The present invention thus concerns a method for increasing the therapeutic index of a bioconjugate and the bioconjugates for use in treatment, in particular cancer. The bioconjugates according to the invention have a sulfamide linker comprising a group according to formula (1).

Bioconjugates containing sulfamide linkers for use in treatment

The present invention relates to a compound comprising an alpha-end and an omega-end, the compound comprising on the alpha-end a reactive group Qcapable of reacting with a functional group Fpresent on a biomolecule and on the omega-end a target molecule, the compound further comprising a group according to formula (1) or a salt thereof: Said compound may also be referred to as a linker-conjugate. The invention also relates to a process for the preparation of a bioconjugate, the process comprising the step of reacting a reactive group Qof a linker-conjugate according to the invention with a functional group Fof a biomolecule. The invention further relates to a bioconjugate obtainable by the process according to the invention.

Sulfamide linker, conjugates thereof, and methods of preparation

The invention concerns fusion proteins, wherein two endoglycosidases are fused, possibly via a linker. The fusion enzymes according to the invention have structure (1): EndoX-(L)-EndoY (1), wherein EndoX is an endoglycosidase, EndoY is an endoglycosidase distinct from EndoX, L is a linker and p is 0 or 1. Such fusion enzymes capable of trimming glycoproteins comprising at least two distinct glycoforms in a single step. The invention further concerns the use of the fusion enzyme according to the invention for trimming glycoproteins. In another aspect, the invention relates to the process of production of the fusion enzyme. In a further aspect, the inventions concerns a process for trimming glycoproteins, comprising trimming the glycoprotein with a fusion enzyme according to the invention, to obtain a trimmed glycoprotein.

Enzymes for trimming of glycoproteins

The present invention concerns novel and improved antibody-conjugates for targeting CD30. The inventors found that when antibody-conjugates were prepared using a specific mode of conjugation, they exhibit an improved therapeutic index. The mode of conjugation comprises a first step (i) of contacting a glycoprotein comprising 1-4 core N-acetylglucosamine moieties with a compound of the formula S(F)-P in the presence of a catalyst, wherein S(F)is a sugar derivative comprising x functional groups Fcapable of reacting with a functional group Q, x is 1 or 2 and P is a nucleoside mono- or diphosphate, and wherein the catalyst is capable of transferring the S(F)moiety to the core-GlcNAc moiety, to obtain a modified antibody; and a second step (ii) of reacting the modified antibody with a linker-conjugate comprising a functional group Qcapable of reacting with functional group Fand a target molecule D connected to Qvia a linker Lto obtain the antibody-conjugate wherein linker L comprises S&#x2014;Z-Land wherein Zis a connecting group resulting from the reaction between Qand F. The invention also relates to a use for improving the therapeutic index of an antibody-conjugate and to a method for targeting CD30-expressing cells.

Antibody-conjugates with improved therapeutic index for targeting CD30 tumours and method for improving therapeutic index of antibody-conjugates

The present invention concerns novel and improved antibody-conjugates for targeting HER2. The inventors found N that when antibody-conjugates were prepared using a specific mode of conjugation, they exhibit an improved therapeutic index. The mode of conjugation comprises a first step (i) of contacting a glycoprotein comprising 1-4 core N-acetylglucosamine moieties with a compound of the formula S(F)-P in the presence of a catalyst, wherein S(F)is a sugar derivative comprising x functional groups F1 capable of reacting with a functional group Q, x is 1 or 2 and P is a nucleoside mono- or diphosphate, and wherein the catalyst is capable of transferring the S(F)moiety to the core-GlcNAc moiety, to obtain a modified antibody; and a second step (ii) of reacting the modified antibody with a linker-conjugate comprising a functional group Qcapable of reacting with functional group Fand a target molecule D connected to Qvia a linker Lto obtain the antibody-conjugate wherein linker L comprises S&#x2014;Z-Land wherein Zis a connecting group resulting from the reaction between Qand F. The invention also relates to a use for improving the therapeutic index of an antibody-conjugate and to a method for targeting HER2-expressing cells.

Antibody-conjugates with improved therapeutic index for targeting HER2 tumours and method for improving therapeutic index of antibody-conjugates

The present invention relates to a process for the enzymatic modification of a glycoprotein. The process comprises the step of contacting a glycoprotein comprising a glycan comprising a terminal GlcNAc-moiety, in the presence of glycosyltransferase that is, or is derived from, a &#x3b2;-(1,4)-N-acetylgalactosaminyltransferase, with a non-natural sugar-derivative nucleotide. The non-natural sugar-derivative nucleotide is according to formula (3): wherein A is selected from the group consisting of &#x2014;N, &#x2014;C(O)R, &#x2014;(CH)C&#x2261;C&#x2014;R, &#x2014;SH, &#x2014;SC(O)R, &#x2014;SC(O)OR, &#x2014;SC(S)OR, &#x2014;F, &#x2014;Cl, &#x2014;Br &#x2014;I, &#x2014;OS(O)R, terminal C-Calkenyl groups, C-Ccycloalkenyl groups, C-Calkadienyl groups, terminal C-Callenyl groups and amino groups. The invention further relates to a glycoprotein obtainable by the process according to the invention, to a bioconjugate that can be obtained by conjugating the glycoprotein with a linker-conjugate, and to &#x3b2;-(1,4)-N-acetylgalactosaminyltransferases that can be used in preparing the glycoprotein according to the invention.

Process for the modification of a glycoprotein using a glycosyltransferase that is or is derived from a &#x3b2;(1,4)-n-acetylgalactosaminyltransferase

The present invention relates to a process for the modification of a glycoprotein, using a &#x3b2;-(1,4)-N-acetylgalactosaminyltransferase or a mutant thereof. The process comprises the step of contacting a glycoprotein comprising a glycan comprising a terminal GlcNAc-moiety, in the presence of a &#x3b2;-(1,4)-N-acetylgalactosaminyltransferase or a mutant thereof, with anon-natural sugar-derivative nucleotide. The non-natural sugar-derivative nucleotideis according to formula (3), wherein A is selected from the group consisting of &#x2014;N; &#x2014;C(O)R; &#x2014;C&#x2550;C&#x2014;R; &#x2014;SH; &#x2014;SC(O)R; &#x2014;SC(V)OR, wherein V is O or S; &#x2014;X wherein X is selected from the group consisting of F, Cl, Br and I; &#x2014;OS(O)R; an optionally substituted C-Calkyl group; an optionally substituted terminal C-Calkenyl group; and an optionally substituted terminal C-Calkenyl group. 

Process for the modification of a glycoprotein using a &#x3b2;-(1,4)-N-acetylgalactosaminyl transferase or a mutant thereof

The invention relates to a novel linker for use in bioconjugates such as antibody-drug-conjugates. The linker according to the invention is represented by formula: (I) wherein: &#x2014;BM is a branching moiety; &#x2014;E is a capping group; &#x2014;SG is a sulfamide group; &#x2014;b, c, d, e, g, i, k, l are independently 0 or 1; &#x2014;f is an integer in the range of 1 to 10; &#x2014;Sp, Sp, Sp, Sp, Spand Spare a spacer moieties; &#x2014;Zand Zare connecting groups. The linker according to the invention is useful in the preparation of linker-conjugates and bioconjugates, and can be used for (a) improving conjugation efficiency in the preparation of the bioconjugate, (b) reducing aggregation during the preparation of the bioconjugate and/or of the bioconjugate, (c) increasing stability of the bioconjugate, and/or (d) increasing therapeutic index of the bioconjugate. 

The present invention relates to a compound comprising an alpha-end and an omega-end, the compound comprising on the alpha-end a reactive group Qcapable of reacting with a functional group Fpresent on a biomolecule and on the omega-end a target molecule, the compound further comprising a group according to formula (1) or a salt thereof:  Said compound may also be referred to as a linker-conjugate. The invention also relates to a process for the preparation of a bioconjugate, the process comprising the step of reacting a reactive group Qof a linker-conjugate according to the invention with a functional group Fof a biomolecule. The invention further relates to a bioconjugate obtainable by the process according to the invention.

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Lent, Netherlands

Sander Sebastiaan Van Berkel