Enhancing the stability and purity and increasing the bioavailability of human milk oligosaccharides or precursors or blends thereof
The present application discloses a method for enhancing the stability of a human milk oligosaccharide (HMO) or a HMO precursor or a HMO blend when stored for extended periods at temperatures above 25° C. by spray-drying an aqueous solution of the HMO or HMO precursor to remove at least 90% of the water and providing a HMO or a HMO precursor or a HMO blend with a specific glass transition temperature. Also disclosed is a method for removing organic solvent residues from a human milk oligosaccharide (HMO) or a HMO precursor or a HMO blend by spray drying. Also disclosed is a method of enhancing the bioavailability of a human milk oligosaccharide (HMO) or a HMO precursor or a HMO blend by spray drying.
1. A method for preparing an amorphous form of a human milk oligosaccharide (HMO) or a HMO blend comprising: dissolving at least one synthetic HMO containing an organic solvent contaminant in water to form an aqueous solution; and spray-drying the aqueous solution to form an amorphous material, wherein the at least one synthetic HMO is selected from the group consisting of lacto-N-triose, lacto-N-tetraose (LNT), lacto-N-neotetraose (LNnT), lacto-N-hexaose (LNH), lacto-N-neohexaose (LNnH), para-lacto-N-hexaose (para-LNH), para-lacto-N-neohexaose (para-LNnH), 2-O-fucosyllactose (2′-FL), 3-O-fucosyllactose (3-FL), 2′,3-di-O-fucosyllactose (DFL), lacto-N-fucopentaose I (LNFP I), lacto-N-fucopentaose II (LNFP II), lacto-N-fucopentaose II (LNFP III), lacto-N-fucopentaose V (LNFP V), fucosyl-lacto-N-neohexaose (F-LNnH), difucosyl-lacto-N-hexaose I (DF-LNH I), difucosyl-lacto-N-hexaose II (DF-LNH II), difucosyl-para-lacto-N-hexaose (DF-para-LNH), difucosyl-para-lacto-N-neohexaose (DF-para-LNnH), 3′-O-sialyllactose (3′-SL), 6′-O-sialyllactose (6′-SL), 3-O-fucosyl-3′-O-sialyllactose (FSL), sialyllacto-N-tetraose a (LST a), sialyllacto-N-tetraose b (LST b), sialyllacto-N-tetraose c (LST c), fucosyl-sialyllacto-N-tetraose a (F-LST a), fucosyl-sialyllacto-N-tetraose b (F-LST b), fucosyl-sialyllacto-N-tetraose c (F-LST c), and disialyllacto-N-tetraose (DS-LNT).
2. The method of claim 1 , wherein the spray-drying results in the removal of at least about 90% of the water and the amorphous form of HMO or HMO blend has a glass transition temperature (Tg) that is at least 40° C.
3. The method of claim 2 , wherein the spray-drying results in the removal of about 90-95% of the water.
4. The method of claim 1 , wherein the at least one synthetic HMO is a crystalline HMO or HMO blend, wherein the stability of the amorphous form of the HMO or HMO blend has a greater stability than the crystalline HMO or HMO blend.
5. The method of claim 1 , wherein the at least one synthetic HMO is selected from the group consisting of lacto-N-tetraose (LNT), lacto-N-neotetraose (LNnT), 2′-O-fucosyllactose (2′-FL), 3-O-fucosyllactose (3-FL), 2′,3-di-O-fucosyllactose (DFL), lacto-N-fucopentaose I (LNFP I), 3′-O-sialyllactose (3′-SL), 6′-O-sialyllactose (6′-SL), 3-O-fucosyl-3′-O-sialyllactose (FSL), sialyllacto-N-tetraose a (LST a), and disialyllacto-N-tetraose (DS-LNT); and the amorphous form of HMO or HMO blend has a glass transition temperature (Tg) that is at least 80° C.
6. The method of claim 5 , wherein the at least one synthetic HMO is selected from the group consisting of lacto-N-tetraose (LNT), lacto-N-neotetraose (LNnT), 2′-O-fucosyllactose (2′-FL), 3-O-fucosyllactose (3-FL), 3′-O-sialyllactose (3′-SL) and 6′-O-sialyllactose (6′-SL).
7. The method of claim 6 , wherein the at least one synthetic HMO is selected from the group consisting of lacto-N-neotetraose (LNnT), 2′-O-fucosyllactose (2′-FL) and 6′-O-sialyllactose (6′-SL).
8. The method of claim 1 , wherein the at least one synthetic HMO comprises two or more HMOs; and the amorphous form of HMO or HMO blend has a glass transition temperature (Tg) that is at least 60° C.
9. The method of claim 8 , wherein the two or more HMOs are selected from the group consisting of lacto-N-tetraose (LNT), lacto-N-neotetraose (LNnT), 2′-O-fucosyllactose (2′-FL), 3-O-fucosyllactose (3-FL), 2′,3-di-O-fucosyllactose (DFL), lacto-N-fucopentaose I (LNFP I), 3′-O-sialyllactose (3′-SL), 6′-O-sialyllactose (6′-SL), 3-O-fucosyl-3′O-sialyllactose (FSL), sialyllacto-N-tetraose a (LST a), and disialyllacto-N-tetraose (DS-LNT).
10. The method of claim 1 , wherein the spray-drying results in the removal of at least 75% of the organic solvent contaminant.
11. The method of claim 1 , wherein the at least one synthetic HMO is obtained via catalytic hydrogenolysis of a protected derivative of HMO; and the organic solvent contaminant is the hydrogenolized protecting group.
12. The method of claim 1 , wherein the at least one synthetic HMO is obtained via catalytic hydrogenolysis of a protected derivative of HMO in a protic solvent; and the organic solvent contaminant is the protic solvent.
13. The method of claim 12 , wherein the protic solvent is a lower alkanol.
14. The method of claim 1 , wherein the at least one synthetic HMO comprises an HMO crystallized from an organic solvent.
15. The method of claim 12 , wherein the HMO crystallized from an organic solvent is 2′-FL crystallized from a lower alkanol.
16. The method of claim 1 , wherein the organic contaminant comprises methanol.
17. The method of claim 16 , wherein the at least one synthetic HMO comprises 2′-O-fucosyllactose (2′-FL).