EP-4739338-A1 - FUNCTIONALIZED DISACCHARIDASE COMPOSITIONS

Abstract

The present invention relates to a composition comprising a solid carrier, a protein disaccharidase or a fragment thereof immobilized on the surface of the solid carrier, a protective layer to protect the protein disaccharidase or a fragment thereof by embedding the protein disaccharidase 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.

Inventors

• LAPRÉVOTTE, Emilie
• BRIAND, Manon
• KRONENBERG, Vanessa
• SPECIOSO, Gabriele
• SHAHGALDIAN, PATRICK
• DUDAL, Yves Victor René

Assignees

• Perseo Pharma AG

Dates

Publication Date

20260513

Application Date

20240704

Claims (20)

1. 1. A composition comprising a solid carrier, a disaccharidase or a fragment thereof immobilized on the surface of the solid carrier, a protective layer to protect the disaccharidase or the fragment thereof by embedding the disaccharidase or the 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 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.
3. 3. The composition according to anyone of claims 1-2, 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.
4. 4. The composition according to anyone of claims 1-3, wherein the functional constituent is immobilized on the surface of the protective layer by non-covalent binding or by covalent binding.
5. 5. The composition according to anyone of claims 1-4, wherein the disaccharidase or a fragment thereof is selected from the goup consisting of lactase or a fragment thereof, maltase or a fragment thereof, isomaltase or a fragment thereof, trehalase or a fragment thereof and invertase or a fragment thereof or mixtures thereof.
6. 6. The composition according to anyone of claims 1-4, wherein the disaccharidase or a fragment thereof is selected from the group consisting of lactase or a fragment thereof and invertase or a fragment thereof or mixtures thereof.
7. 7. The composition according to anyone of claims 1-4, wherein the disaccharidase or a fragment thereof is invertase or a fragment thereof
8. 8. The composition of anyone of claims 1-7, wherein the protective layer embeds the solid carrier and embeds the disaccharidase or a fragment thereof immobilized on the surface of the solid carrier.
9. 9. The composition of anyone of claims 1-8, wherein the functional constituent immobilized on the surface of the protective layer is not embedded by the protective layer.
10. 10. The composition of anyone of claims 1-9, for use as a medicament.
11. 11. The composition of anyone of claims 1-9, for use in a method of treatment of lactase deficiency, sucrase-isomaltase deficiency and disaccharidoses intolerances.
12. 12. A method of producing a composition, the composition comprising a solid carrier, disaccharidase or a fragment thereof immobilized on the surface of the solid carrier, a protective layer to protect the disaccharidase or the fragment thereof by embedding the disaccharidase or the 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) immobilizing a disaccharidase or a fragment thereof on the solid carrier; (c) forming a protective layer on the surface of the solid carrier to protect the disaccharidase or the fragment thereof immobilized on the solid carrier; (d) 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.
13. 13. The method of claim 12, wherein in step (b), i) a linker is added to the solid carrier provided in step (a), and ii) the disaccharidase or a fragment thereof is added to the solid carrier and the linker, wherein the linker connects the solid carrier with the disaccharidase or a fragment thereof.
14. 14. The method of claim 13, wherein the linker which has not connected the solid carrier with the disaccharidase or a fragment thereof in step (b), is present during formation of a protective layer on the surface of the solid carrier in step (c).
15. 15. The method of claim 13, wherein there is no washing step between adding the linker to the solid carrier provided in step (a) in (i) and adding the disaccharidase or a fragment thereof to the solid carrier and the linker in ii).
16. 16. The method of anyone of claims 12-15, wherein there is no washing step between any of steps (a) to (c).
17. 17. The method of anyone of claims 13-16, wherein the linker which has not connected the solid carrier with the disaccharidase or a fragment thereof in step (b), or a part thereof, covalently binds the protective layer to the disaccharidase or the fragment thereof in step (c).
18. 18. The method of any one of claims 13-17, wherein the linker is selected from the group consisting of glutaraldehyde, disuccinimidyl tartrate, bis[sulfosuccinimidyl]suberate, ethylene glycolbis(sulfosuccinimidylsuccinate), dimethyl adipimidate, dimethyl pimelimidate, sulfosuccinimidyl (4-iodoacetyl) aminobenzoate, l,5-difluoro-2,4-dinitrobenzene, BSOCOES (Bis[2- (succinimidooxycarbonyloxy)ethyl]sulfone), DSP (Dithiobis[succinimidyl]propionate]), DTSSP (3,3 '- Dithiobis[sulfosuccinimidyl]propionate]), DTBP (Dimethyl 3,3 '- dithiobispropionimidate-2 HC1), DST (Disuccinimidyl tartarate), BMDB (1,4 bismaleimidyl-2,3-dihydroxybutane).
19. 19. The method of any one of claims 13-17, wherein the linker is glutaraldehyde.
20. 20. A composition comprising a solid carrier, an disaccharidase or a fragment thereof immobilized on the surface of the solid carrier, a protective layer to protect the disaccharidase or the fragment thereof by embedding the disaccharidase or the 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, wherein the composition is obtainable by the method of any one of claims 14-19.

Description

Functionalized disaccharidase compositions The field of the invention The present invention relates to a composition comprising a solid carrier, a disaccharidase or a fragment thereof immobilized on the surface of the solid carrier, a protective layer to protect the disaccharidase or a fragment thereof by embedding the disaccharidase 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. Background of the invention Disaccharides are normally split into monosaccharides by disaccharidases located in the brush border of small-bowel enterocytes. Undigested disaccharides cause an osmotic load that attracts water and electrolytes into the bowel, causing watery diarrhea. Bacterial fermentation of carbohydrates in the colon produces gases (hydrogen, carbon dioxide, and methane), resulting in excessive flatus, bloating and distention, and abdominal pain. Diseases which are related to the defective digestion of disaccharides are e.g. lactase deficiency and congenital sucrase-isomaltase deficiency (CSID). In case of lactase deficiency, lactose malabsorption is attributable to an imbalance between the amount of ingested lactose and the capacity for lactase to hydrolyze the disaccharide. It manifests as lactose intolerance, a clinical syndrome of one or more of the following symptoms: abdominal pain, diarrhea, nausea, flatulence, and/or bloating after the ingestion of lactose or lactose-containing food substances. The most common cause is primary (hereditary) lactase deficiency attributable to relative or absolute absence of lactase that develops in childhood at various ages in different racial groups. Secondary (acquired) lactase deficiency results from small bowel injury, such as acute gastroenteritis, persistent diarrhea, small bowel overgrowth, cancer chemotherapy, or other causes of injury to the small intestinal mucosa. Congenital lactase deficiency is an extremely rare, autosomal recessive enzyme defect that prevents lactase expression from birth, requiring complete lactose avoidance. Besides lactose avoidance, which could cause significant health issues , enzyme-replacement therapy with microbial exogenous lactase (P-galactosidase obtained from Aspergillus oryzae) prior to or added to dairy meals represents a possible strategy for primary and secondary lactase deficiency. Results about the exact rate of efficacy are discordant. CSID is a multifaceted malabsorption disorder of the small intestine with an autosomal recessive mutation in the sucrase-isomaltase gene, characterized by complete, or almost complete lack of sucrose activity and varying degrees of reduction in isomaltase activity. When this enzyme complex is deficient, nutrients from ingested starch and sucrose cannot be absorbed sufficiently and clinically manifests with diarrhea, abdominal pain and bloating, leading to malnutrition, especially in children. Besides life-long dietary measures, the only effective treatment option is the enzyme replacement with sacrosidase, currently approved in the U.S. by FDA (Sucraid®), but not in Europe. Thus there is a need to provide effective treatments for diseases which are related to the defective digestion of disaccharides. Summary of the invention The present invention provides a composition comprising a solid carrier, a disaccharidase or a fragment thereof immobilized on the surface of the solid carrier, a protective layer to protect the disaccharidase or a fragment thereof by embedding the disaccharidase 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 provides also a method of producing said composition comprising a solid carrier, a disaccharidase or a fragment thereof immobilized on the surface of the solid carrier, a protective layer to protect the disaccharidase or a fragment thereof by embedding the disaccharidase 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) immobilizing a disaccharidase or a fragment thereof on the solid carrier; (c) forming a protective layer on the surface of the solid carrier to protect the disaccharidase or a fr