IP Library › Granted Patent US 12,656,341
Granted Patent B2
US 12,656,341 · App. 17/438,247 · Granted Jun 16, 2026

Process for the preparation of lipidated proteinaceous structures

Inventors: Steffen Woell (Mainz, DE); Stefan Schiller (Mainz, DE); Simon Geissler (Bad Homburg, DE)
Assignee: MERCK PATENT GMBH
G01N33/5432A61K47/65A61K47/6891
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Quick Facts
Patent No.
US 12,656,341
App. No.
17/438,247
Granted
Jun 16, 2026
Kind
B2
Abstract

The present invention relates to a process for the preparation of a conjugate, comprising a biological molecule, an enzymatic tag, a hydrophilic spacer, a linker and a lipophilic moiety using enzymatic coupling.

Claims (27)

1 . A process for the preparation of a conjugate, suitable to be inserted into hydrophobic environments selected from the group consisting of hydrophobic polymeric surfaces, lipid based drug delivery systems and membranes of living cells, wherein the conjugate comprises a biological molecule, an enzymatic tag, a hydrophilic spacer, a linker and a lipophilic moiety, wherein such process comprises enzymatic coupling of a component comprising the enzymatic tag, the hydrophilic spacer, the linker and the lipophilic moiety with the biological molecule in an aqueous medium and wherein such process comprises the following steps:

a) preparing an aqueous dispersion of a component which has the formula (II):

wherein

m is any integer number from 15 to 60;

n is any integer number from 3 to 27;

p is any integer number from 0 to 9;

b) adding a transpeptidase and the biological molecule which is a polypeptide selected from an antigen, a cell adhesion protein, a peptide hormone, a cytokine, or a receptor related to any of these molecules, an enzyme, or a natural or artificial antibody or fragment thereof;

c) incubating the mixture obtained in step (b) to produce the conjugate with the biological molecule;

d) purifying the conjugate obtained in step (c).

2 . The process according to claim 1 , wherein the antibody is a monoclonal antibody or a fragment thereof, such as a single-chain variable fragment (scFv), a variable fragment (Fv), or a fragment antigen binding (Fab, Fab′ or F (ab′) 2); a camelid or cartilaginous fish-derived heavy-chain only antibody or a fragment thereof, such as a VHH or a vNAR, or wherein the artificial antibody is a DARPin, a adnectine, an anticalin, or an affibody.

3 . The process according to claim 1 , wherein the biological molecule is a single-domain antibody derived from the variable domain of camelid heavy-chain only antibodies (VHH).

4 . The process according to claim 1 , wherein the biological molecule prior to its coupling with the component according to formula (II) carries a C-terminal motif for enzymatic conjugation by transpeptidases.

5 . The process according to claim 4 , wherein the C-terminal motif consists of the amino acid sequence “leucine-proline-X-threonine-glycine” (LPXTG), wherein “X” can be any proteinogenic amino acid.

6 . The process according to claim 5 , wherein the proteinogenic amino acid present in the LPXTG motif is glycine, alanine, valine, leucine, isoleucine, methionine, phenylalanine, proline, serine, threonine, tyrosine, asparagine, glutamine, aspartic acid, glutamic acid, lysine, arginine or histidine.

7 . The process according to claim 1 , wherein the component according to formula (II) is

8 . The process according to claim 1 , wherein the biological molecule is an integrin, a cadherin, a growth factor or an interleukin.

9 . The process according to claim 1 , wherein m is from 25 to 45.

10 . The process according to claim 9 , wherein m is from 30 to 40.

11 . The process according to claim 10 , wherein m is 36.

12 . The process according to claim 1 , wherein n is from 7 to 19.

13 . The process according to claim 12 , wherein n is from 11 to 15.

14 . The process according to claim 13 , wherein n is 11.

15 . The process according to claim 1 , wherein p is from 2 to 7.

16 . The process according to claim 15 , wherein p is from 3 to 5.

17 . The process according to claim 16 , wherein p is 4.

18 . The process according to claim 1 wherein the transpeptidase in step (b) is sortase.

19 . The process according to claim 18 wherein the sortase is sortase A.

Assignments (2)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Oct 7, 2022
From: WOELL, STEFFEN; SCHILLER, STEFAN; GEISSLER, SIMON
To: MERCK HEALTHCARE KGAA
Reel/Frame 061345/0929 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Oct 7, 2022
From: MERCK HEALTHCARE KGAA
To: MERCK PATENT GMBH
Reel/Frame 061346/0006 →
Priority Claims (1)
EP 19162511 · Mar 13, 2019 · regional
Continuity (1)
Related Publication 20220184223A1 · Jun 16, 2022
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