EP-4739337-A1 - FUNCTIONALIZED BIOCATALYTICAL COMPOSITIONS COMPRISING PANCREATIC ENZYMES

Abstract

The present invention relates to a composition comprising a solid carrier, a lipase or a fragment thereof immobilized on the surface of the solid carrier wherein the lipase or a fragment thereof is in the open conformation, a protease or a fragment thereof immobilized on the surface of the solid carrier, an amylase or a fragment thereof immobilized on the surface of the solid carrier, an agent which interacts with the lid domain of the lipase or a fragment thereof, a protective layer to protect the lipase or a fragment thereof, the protease or a fragment thereof and the amylase or a fragment thereof by embedding the lipase or a fragment thereof, the protease or a fragment thereof and the amylase or a fragment thereof, and a functional constituent immobilized on the surface of the protective layer, wherein the functional constituent immobilized on the surface of the protective layer is a polymer comprising repeat units wherein each repeat unit comprises at least one amino group and/or at least one thiol group.

Inventors

• LAPRÉVOTTE, Emilie
• BRIAND, Manon
• DUDAL, Yves Victor René
• SPECIOSO, Gabriele
• SHAHGALDIAN, PATRICK
• WATSON, John Ashleigh

Assignees

• Perseo Pharma AG
• Société des Produits Nestlé S.A.

Dates

Publication Date

20260513

Application Date

20240704

Claims (20)

1. 1. A composition comprising a solid carrier, a lipase or a fragment thereof immobilized on the surface of the solid carrier wherein the lipase or a fragment thereof is in the open conformation, a protease or a fragment thereof immobilized on the surface of the solid carrier, an amylase or a fragment thereof immobilized on the surface of the solid carrier, an agent which interacts with the lid domain of the lipase or a fragment thereof, a protective layer to protect the lipase or a fragment thereof, the protease or a fragment thereof and the amylase or a fragment thereof by embedding the lipase or a fragment thereof, the protease or a fragment thereof and the amylase or a fragment thereof, and a functional constituent immobilized on the surface of the protective layer, wherein the functional constituent immobilized on the surface of the protective layer is a polymer comprising repeat units wherein each repeat unit comprises at least one amino group and/or at least one thiol group.
2. 2. The composition according to claim 1, wherein the agent which interacts with the lid domain of the lipase or a fragment thereof is selected from the group consisting of a colipase or a fragment thereof, a colipase-mimicking peptide, and an amphipathic molecule.
3. 3. The composition according to claim 1 or 2, wherein the agent interacts specifically with the lid domain of the lipase or a fragment thereof so that the lipase or a fragment thereof shifts to and/or maintains the open conformation.
4. 4. The composition according to any one of claims 1-3, wherein about 100% of the lipase or a fragment thereof immobilized on the surface of the solid carrier is in the open conformation.
5. 5. The composition according to any one of claims 1-4, wherein the polymer comprising repeat units wherein each repeat unit comprises at least one amino group and/or at least one thiol group is a polyglucosamin selected from the group consisting of chitin, chitosan, polyglucosaminoglycans, chondroitin, heparin, keratan and dermatan or a derivative thereof.
6. 6. The composition according to any one of claims 1-4, wherein the polymer comprising repeat units wherein each repeat unit comprises at least one amino group and/or at least one thiol group is chitosan or a derivative thereof.
7. 7. The composition according to any one of claims 1-6, wherein the functional constituent is immobilized on the surface of the protective layer by non-covalent binding or by covalent binding.
8. 8. The composition according to any one of claims 1-7, wherein the composition comprises a solid carrier; pancreatin, wherein the pancreatin comprises a lipase or a fragment thereof wherein the lipase or a fragment thereof is in the open conformation, a protease or a fragment thereof and an amylase or a fragment thereof, wherein the lipase or a fragment thereof, the protease or a fragment thereof and the amylase or a fragment thereof are immobilized on the surface of the solid carrier; an agent which interacts with the lid domain of the lipase or a fragment thereof; a protective layer to protect the lipase or a fragment thereof, the protease or a fragment thereof and the amylase or a fragment thereof by embedding the lipase or a fragment thereof, the protease or a fragment thereof and the amylase or a fragment thereof; and a functional constituent immobilized on the surface of the protective layer, wherein the functional constituent immobilized on the surface of the protective layer is a polymer comprising repeat units wherein each repeat unit comprises at least one amino group and/or at least one thiol group.
9. 9. The composition of any one of claims 1-8, wherein the protective layer embeds the solid carrier, embeds the lipase or a fragment thereof immobilized on the surface of the solid carrier, embeds the protease or a fragment thereof immobilized on the surface of the solid carrier and embeds the amylase or a fragment thereofimmobilized on the surface of the solid carrier.
10. 10. The composition of any one of claims 1-9, wherein the functional constituent immobilized on the surface of the protective layer is not embedded by the protective layer.
11. 11. The composition of any one of claims 1-10, for use as a medicament.
12. 12. The composition of any one of claims 1-10, for use in a method of enzyme replacement therapy (ERT) preferably gastrointestinal enzyme replacement therapy or for use in a method for the prevention, delay of progression or treatment of exocrine pancreatic insufficiency (EPI).
13. 13. A method of producing a composition, the composition comprising a solid carrier, a lipase or a fragment thereof immobilized on the surface of the solid carrier wherein the lipase or a fragment thereof is in the open conformation, a protease or a fragment thereof immobilized on the surface of the solid carrier, an amylase or a fragment thereof immobilized on the surface of the solid carrier, an agent which interacts with the lid domain of the lipase or a fragment thereof, a protective layer to protect the lipase or a fragment thereof, the protease or a fragment thereof and the amylase or a fragment thereof by embedding the lipase or a fragment thereof, the protease or a fragment thereof and the amylase or a fragment thereof, and a functional constituent immobilized on the surface of the protective layer, wherein the functional constituent immobilized on the surface of the protective layer is a polymer comprising repeat units wherein each repeat unit comprises at least one amino group and/or at least one thiol group, the method comprising the following steps: (a) providing a solid carrier; (b) providing a lipase or a fragment thereof, a protease or a fragment thereof and an amylase or a fragment thereof; (c)providing an agent which interacts with the lid domain of a lipase or a fragment thereof; (d) allowing the lipase or a fragment thereof of (b) to interact with the agent of (c); (e) immobilizing the lipase or a fragment thereof, the protease or a fragment thereof and the amylase or a fragment thereof on the solid carrier; (f) forming a protective layer on the surface of the solid carrier to protect the lipase or a fragment thereof, the protease or a fragment thereof and the amylase or a fragment thereof immobilized on the solid carrier; (g) immobilizing a functional constituent on the surface of the protective layer, wherein the functional constituent immobilized on the surface of the protective layer is a polymer comprising repeat units, wherein each repeat unit comprises at least one amino group and/or at least one thiol group.
14. 14. A method of producing a composition, the composition comprising a solid carrier, a lipase or a fragment thereof immobilized on the surface of the solid carrier wherein the lipase or a fragment thereof is in the open conformation, a protease or a fragment thereof immobilized on the surface of the solid carrier, an amylase or a fragment thereof immobilized on the surface of the solid carrier, an agent which interacts with the lid domain of the lipase or a fragment thereof, a protective layer to protect the lipase or a fragment thereof, the protease or a fragment thereof and the amylase or a fragment thereof by embedding the lipase or a fragment thereof, the protease or a fragment thereof and the amylase or a fragment thereof , and a functional constituent immobilized on the surface of the protective layer, wherein the functional constituent immobilized on the surface of the protective layer is a polymer comprising repeat units wherein each repeat unit comprises at least one amino group and/or at least one thiol group, the method comprising the following steps: (a) providing a solid carrier; (b) providing a lipase or a fragment thereof, a protease or a fragment thereof and an amylase or a fragment thereof; (c) immobilizing the lipase or a fragment thereof, the protease or a fragment thereof and the amylase or a fragment thereof on the solid carrier; (d) providing an agent which interacts with the lid domain of a lipase or a fragment thereof; (e) allowing the lipase or a fragment thereof of (b) to interact with the agent of (d); (f) forming a protective layer on the surface of the solid carrier to protect the lipase or a fragment thereof, the protease or a fragment thereof and the amylase or a fragment thereof immobilized on the solid carrier; (g) immobilizing a functional constituent on the surface of the protective layer, wherein the functional constituent immobilized on the surface of the protective layer is a polymer comprising repeat units, wherein each repeat unit comprises at least one amino group and/or at least one thiol group.
15. 15. The method of claim 14 or 15, wherein in step b) pancreatin comprising a lipase or a fragment thereof, a protease or a fragment thereof and an amylase or a fragment thereof is provided.
16. 16. The method of claim 13, wherein i) a linker is added to the solid carrier provided in step (a), and ii) the lipase or a fragment thereof, the protease or a fragment thereof and the amylase or a fragment thereof provided in step (b) is added to the solid carrier and the linker, wherein the linker connects the solid carrier with the lipase or a fragment thereof, the protease or a fragment thereof and the amylase or a fragment thereof in step (e).
17. 17. The method of claim 16, wherein the linker which has not connected the solid carrier with the lipase or a fragment thereof, the protease or a fragment thereof and the amylase or a fragment thereof in step (e), is present during formation of a protective layer on the surface of the solid carrier in step (f).
18. 18. The method of claim 16, wherein there is no washing step between adding the linker to the solid carrier provided in step (a) in (i) and adding the lipase or a fragment thereof, the protease or a fragment thereof and the amylase or a fragment thereof to the solid carrier and the linker in ii).
19. 19. The method of any one of claims 13-18, wherein there is no washing step between any of steps (a) to (f).
20. 20. The method of any one of claims 13-19, wherein the linker which has not connected the solid carrier with the lipase or a fragment thereof, the protease or a fragment thereof and the amylase or a fragment thereof in step (e), or a part thereof, covalently binds the protective layer to the lipase or a fragment thereof, the protease or a fragment thereof and the amylase or a fragment thereof in step (f).

Description

Functionalized biocatalytical compositions comprising pancreatic enzymes The field of the invention The present invention relates to a composition comprising a solid carrier, a lipase or a fragment thereof immobilized on the surface of the solid carrier wherein the lipase or a fragment thereof is in the open conformation, a protease or a fragment thereof immobilized on the surface of the solid carrier, an amylase or a fragment thereof immobilized on the surface of the solid carrier, an agent which interacts with the lid domain of the lipase or a fragment thereof, a protective layer to protect the lipase or a fragment thereof, the protease or a fragment thereof and the amylase or a fragment thereof by embedding the lipase or a fragment thereof, the protease or a fragment thereof and the amylase or a fragment thereof, and a functional constituent immobilized on the surface of the protective layer, wherein the functional constituent immobilized on the surface of the protective layer is a polymer comprising repeat units wherein each repeat unit comprises at least one amino group and/or at least one thiol group. The present invention also relates to methods of producing said composition and uses thereof. Background of the invention Exocrine Pancreatic Insufficiency (EPI) is a condition in which the exocrine functions of the pancreas are impaired and its ability to effectively deliver digestive enzymes to the duodenum is defective. It is a serious health condition that leads to malabsorption of lipids, fat-soluble vitamins, proteins, and, to a lesser extent, carbohydrates from the digestive tract. The standard medical therapy to treat clinical symptoms and malabsorption is oral pancreatic enzyme replacement therapy (PERT). Current approved therapies consist in pancreatic enzyme product (lipase, amylase, protease) from porcine origin, also called pancreatin. A minimum dose of 40 000 - 50 000 Units of PERT -lipase is recommended at each main meals and half that dose with snacks. Such doses lead to a heavy pill burden for the patients. However, the current treatment is not efficient and exhibits several limitations: Inefficiency of the treatment, persistence of the symptoms, low intraluminal survival time of the lipase that is susceptible to proteases degradation (default of fat digestion), possible intolerance to massive loads of enzymes, and/or gastrointestinal troubles due to the high amount of proteases ingested. Thus, there is a need to provide compatible and effective compositions for treatment of Exocrine Pancreatic Insufficiency (EPI). Summary of the invention The present invention provides a composition comprising a solid carrier, a lipase or a fragment thereof immobilized on the surface of the solid carrier wherein the lipase or a fragment thereof is in the open conformation, a protease or a fragment thereof immobilized on the surface of the solid carrier, an amylase or a fragment thereof immobilized on the surface of the solid carrier, an agent which interacts with the lid domain of the lipase or a fragment thereof, a protective layer to protect the lipase or a fragment thereof, the protease or a fragment thereof and the amylase or a fragment thereof by embedding the lipase or a fragment thereof, the protease or a fragment thereof and the amylase or a fragment thereof , and a functional constituent immobilized on the surface of the protective layer, wherein the functional constituent immobilized on the surface of the protective layer is a polymer comprising repeat units wherein each repeat unit comprises at least one amino group and/or at least one thiol group. The present invention also provides a method of producing a composition, the composition comprising a solid carrier, a lipase or a fragment thereof immobilized on the surface of the solid carrier wherein the lipase or a fragment thereof is in the open conformation, a protease or a fragment thereof immobilized on the surface of the solid carrier, an amylase or a fragment thereof immobilized on the surface of the solid carrier, an agent which interacts with the lid domain of the lipase or a fragment thereof, a protective layer to protect the lipase or a fragment thereof, the protease or a fragment thereof, and the amylase or a fragment thereof by embedding the lipase or a fragment thereof, the protease or a fragment thereof and the amylase or a fragment thereof, and a functional constituent immobilized on the surface of the protective layer, wherein the functional constituent immobilized on the surface of the protective layer is a polymer comprising repeat units wherein each repeat unit comprises at least one amino group and/or at least one thiol group, the method comprising the following steps: (a) providing a solid carrier; (b) providing a lipase or a fragment thereof, a protease or a fragment thereof and an amylase or a fragment thereof ; (c) providing an agent which interacts with the lid domain of a lipase or a fragment thereof; (d) allowing t