Hydrated and anhydrous polymorphs of 2′-O-fucosyllactose and their production methods
This invention describes new hydrated and anhydrous polymorphs of 2′-0-fucosyllactose (2′FL): Polymorph A 2′FL-3/2H 2 O, Polymorph B 2TL-5/2 H 2 O and anhydrous Polymorph C. There is also a description of the methods for obtaining them, and of a new method for preparing Polymorph I already known in the literature.
1. A crystalline 2′-O-Fucosyllactose (2′FL)hydrate with molecular formula C 18 H 32 O 15 ·nH 2 O wherein n is 3/2 in form of polymorph A, wherein said polymorph A is 2′FL·3/2H 2 O, having characteristic XRPD peaks at 18.86±0.20, 9.89±0.20, 17.05±0.20 2θ.
2. The crystalline 2′FL according to claim 1 , having further characteristic XRPD peaks at 21.65±0.2, and 14.20±0.20 2θ.
3. The crystalline 2′FL according to claim 1 , wherein a single crystal of said crystalline 2′FL has a P2 1 2 1 2 1 spatial group which is arranged within an orthorhombic crystalline system and said crystal comprises elementary cells having the following parameters: a=12.4098(8) Å, b=12.737(2) Å, c=13.756(2) Å and a volume of 2212.5 Å 3 .
4. The crystalline 2′FL according to claim 1 , having β anomeric conformation.
5. A pharmaceutical or nutritional or cosmetic composition comprising 2′FL hydrate according to claim 1 and at least one other pharmaceutically, cosmetic or nutritionally acceptable ingredient.
6. A method for obtaining the 2′-O-Fucosyllactose (2′FL) hydrate of claim 1 , comprising preparing an aqueous solution of 2′FL at a concentration by weight between 53% and 75%, and a temperature between 50° C. and 60° C.;
adding at least one warm solvent or a solvent at boiling temperature to obtain a 2′FL crystallization mixture, wherein said warm solvent or said solvent at boiling temperature is selected from the group consisting of methanol, ethanol, propanol, acetone acetonitrile and acetic acid; further comprising keeping said 2′FL crystalline mixture and said solvent at a temperature between 50° C. and 60° C.;
heating said 2′FL crystallization mixture to a temperature between 55° C. and 65° C. for 1-16 hours to obtain formation of crystallization nuclei and allowing the 2′FL crystallization mixture to cool at room temperature to obtain polymorph A via precipitation; and
isolating precipitated polymorph A by filtration, washing and drying.
7. The method according to claim 6 , wherein the warm solvent or solvent at boiling temperature is selected from the group consisting of methanol, ethanol, and propanol.
8. The method according to claim 6 , wherein said warm solvent or solvent at boiling temperature added to said aqueous solution of 2′FL contains between 10 and 40% (v/v) of water.
9. A crystalline 2′-O-Fucosyllactose (2′FL) hydrate with molecular formula C 18 H 32 O 15 .nH 2 O wherein n is 5/2, in form of polymorph B, wherein said polymorph B is 2′FL.5/2 H 2 O, having characteristic XRPD peaks at 9.96±0.20, 18.56±0.2, and 20.48+0.20 2θ.
10. The polymorph B according to claim 9 , having further characteristic XRPD peaks at 14.21±0.20, and 11.90±0.20 2θ.
11. The polymorph B according to claim 9 , wherein single crystal of said crystalline 2′FL has a P2 1 2 1 2 1 spatial group which is arranged within an orthorhombic crystalline system and said crystal comprises elementary cells having the following parameters: a=14.905(7) Å, b=12.663(6) Å, c=12.447(5) Å and a volume of 2349.4 Å 3 .
12. The polymorph B according to claim 9 , having β anomeric conformation.
13. A pharmaceutical or nutritional or cosmetic composition comprising 2′FL hydrate according to claim 9 and at least one other pharmaceutically, cosmetic or nutritionally acceptable ingredient.
14. A method for obtaining polymorph B according to claim 9 , said method comprising
(a) preparing a warm aqueous solution of 2′FL, said 2′FL present in said solution at a concentration of 50-80% by weight,
(b) keeping the aqueous solution at 30-60° C. for 1-12 hours,
(c) cooling the solution to a temperature between 20 and 30° C. for a period of time from 1 hours to 10 days to obtain polymorph B via precipitation, followed by;
(d) cooling the mixture to a temperature between 2 and 18° C. and;
(e) collecting precipitated polymorph B by filtration, washing and drying; said washing being carried out using water at 4-15° C.
15. The method of claim 14 , further comprising seeding the aqueous solution of (b) with 2′FL polymorph B to facilitate formation of polymorph B crystals.