Oligosaccharide compositions and methods of use thereof
Aspects of the disclosure relate to oligosaccharide compositions and methods of making the same. Also provided are methods of using oligosaccharide compositions as microbiome metabolic therapies for reducing ammonia levels and for the treatment of related diseases.
1. An oligosaccharide composition comprising a plurality of oligosaccharides that comprise Formula (I) and Formula (II):
wherein R in Formula (I) and Formula (II) is independently selected from hydrogen, and Formulae (Ia), (Ib), (Ic), (Id), (IIa), (IIb), (IIc), (IId):
wherein R in Formulae (Ia), (Ib), (Ic), (Id), (IIa), (IIb), (IIc), and (IId) is independently defined as above in Formula (I) and Formula (II);
wherein the oligosaccharide composition is produced by a process comprising:
(a) forming a reaction mixture comprising a dextrose preparation and a galactose preparation with a catalyst comprising positively charged hydrogen ions; and
(b) promoting acid catalyzed oligosaccharide formation in the reaction mixture by transferring sufficient heat to the reaction mixture to maintain the reaction mixture at its boiling point until the weight percent of dextrose monomer and galactose monomer in the oligosaccharide composition is in a range of 5%-20% w:w; and
wherein the plurality of oligosaccharides is characterized by a multiplicity-edited gradient-enhanced 1 H- 13 C heteronuclear single quantum correlation (HSQC) NMR spectrum comprising signals 1 to 11:
Center Position
(ppm)
Area under the curve (AUC)
Signal
1 H
13 C
(% of total areas of signals 1-11)
1
3.67
63.35
18.67-26.54
2
3.97
66.25
4.46-7.34
3
3.88
67.13
2.39-4.52
4
3.71
67.34
6.06-8.68
5
3.83
71.27
5.18-8.84
6
3.96
71.58
5.40-8.38
7
3.72
72.4
5.57-11.87
8
3.33
73.76
12.44-19.14
9
4.06
77.39
4.19-9.48
10
4.11
81.73
4.01-6.81
11
4.51
103.34
7.98-11.84.
2. The oligosaccharide composition of claim 1 , wherein step (b) further comprises removing water from the reaction mixture by evaporation.
3. The oligosaccharide composition of claim 1 , wherein the reaction mixture is maintained at its boiling point until the molar ratio of net water condensate produced by the reaction mixture relative to the total dextrose and galactose in the dextrose and galactose preparations prior to loading in (a) is in a range of 0.1-1.0.
4. The oligosaccharide composition of claim 1 , wherein, prior to step (b), the temperature of the reaction mixture is gradually increased from room temperature to the boiling point of the reaction mixture under suitable conditions to achieve homogeneity and uniform heat transfer.
5. The oligosaccharide composition of claim 1 , wherein the reaction mixture in step (a) comprises a molar ratio of dextrose to galactose of about 1:1.
6. The oligosaccharide composition of claim 1 , wherein the process further comprising:
(c) quenching the reaction mixture using water while bringing the temperature of the reaction mixture to below 100° C.
7. The oligosaccharide composition of claim 6 , wherein in step (c) the water has a temperature of about 60-100° C.
8. The oligosaccharide composition of claim 7 , wherein the process further comprises: (d) separating at least a portion of the oligosaccharides from the catalyst.
9. The oligosaccharide composition of claim 8 , wherein the process further comprises one or more of steps (e) to (g):
(e) passing the diluted composition through a cationic exchange resin;
(f) passing the diluted composition through an anionic exchange resin; and
(g) passing the diluted composition through a decolorizing polymer resin;
wherein each of (e), (f), and (g) can be performed one or more times in any order.
10. The oligosaccharide composition of claim 1 , wherein the catalyst is a soluble acid catalyst.
11. The oligosaccharide composition of claim 10 , wherein the soluble acid catalyst is citric acid.
12. The oligosaccharide composition of claim 1 , wherein the oligosaccharide composition has been subjected to a demonomerization process, optionally wherein the oligosaccharide composition comprises less than 2% monomer.
13. The oligosaccharide composition of claim 1 , wherein less than 10% of the oligosaccharide composition or the plurality of oligosaccharides comprise monomers or oligomer subunits that are different from Formula (I), Formula (II), or Formulae (Ia), (Ib), (Ic), (Id), (IIa), (IIb), (IIc), and (IId).
14. The oligosaccharide composition of claim 1 , wherein about 75 to 95 weight percent (dry basis) of the oligosaccharide composition comprises oligosaccharides having a degree of polymerization of two or more monomers (DP2+).
15. The oligosaccharide composition of claim 1 , wherein the oligosaccharide composition comprises less than 15% monomer (DP1).
16. The oligosaccharide composition of claim 1 , wherein the oligosaccharide composition comprises less than 3% w/w impurities.
17. The oligosaccharide composition of claim 16 , wherein the impurities comprise levoglucosan, lactic acid, and/or formic acid.
18. The oligosaccharide composition of claim 1 , wherein the plurality of oligosaccharides comprises 55% to 95% total dietary fiber (dry basis).
19. The oligosaccharide composition of claim 1 , wherein any or all of NMR signals 1-11 are defined as follows:
Center Position
(ppm)
Area under the curve (AUC)
Signal
1 H
13 C
(% of total areas of signals 1-11)
1
3.67
63.35
20.25-24.97
2
3.97
66.25
5.03-6.76
3
3.88
67.13
2.82-4.09
4
3.71
67.34
6.59-8.15
5
3.83
71.27
5.91-8.11
6
3.96
71.58
5.99-7.78
7
3.72
72.4
6.83-10.61
8
3.33
73.76
13.78-17.80
9
4.06
77.39
5.25-8.42
10
4.11
81.73
4.57-6.25
11
4.51
103.34
8.75-11.07.
20. The oligosaccharide composition of claim 1 , wherein signals 1-11 are each further characterized by an 1 H integral region and a 13 C integral region, defined as follows:
1 H Position (ppm)
13 C Position (ppm)
Center
1 H Integral Region
Center
13 C Integral Region
Signal
Position
from
To
Position
from
to
1
3.67
3.607
3.742
63.35
63.78
62.92
2
3.97
3.925
4.008
66.25
66.78
65.73
3
3.88
3.848
3.910
67.13
67.50
66.76
4
3.71
3.674
3.753
67.34
68.02
66.65
5
3.83
3.791
3.864
71.27
71.70
70.84
6
3.96
3.906
4.014
71.58
71.91
71.24
7
3.72
3.669
3.777
72.4
72.78
72.02
8
3.33
3.262
3.404
73.76
74.29
73.23
9
4.06
4.022
4.108
77.39
77.89
76.90
10
4.11
4.066
4.147
81.73
82.15
81.32
11
4.51
4.461
4.556
103.34
103.95
102.72
21. The oligosaccharide composition of claim 1 , wherein the NMR spectrum is obtained by subjecting a sample of the composition to a multiplicity-edited gradient-enhanced 1 H- 13 C heteronuclear single quantum coherence (HSQC) experiment using an echo-antiecho scheme for coherence selection using the pulse sequence diagram as set forth in FIG. 10 ; and the following acquisition parameters and processing parameters:
(i) 1 H Carrier Frequency=4 ppm;
(ii) 13 C Carrier Frequency=65 ppm;
(iii) Number of points in acquisition dimension=596;
(iv) Spectral range in acquisition dimension=6.00 ppm to 2.03 ppm;
(v) Number of points in indirect dimension=300 complex points;
(vi) Spectral range in indirect dimension=120 ppm to 10 ppm;
(vii) Recycle delay=1 second;
(viii) One-bond 1 H- 13 C coupling constant=J CH =146 Hz;
(ix) Number of scans=8;
(x) Temperature=298 K;
(xi) Solvent=D 2 O;
(xii) Window function in direct dimension=Gaussian broadening, 7.66 Hz;
(xiii) Window function in indirect dimension=Gaussian broadening 26.48 Hz; and
(xiv) Processing=512 complex points in direct dimension, 1024 complex points in indirect dimension.
22. The oligosaccharide composition of claim 1 , wherein the NMR spectrum is obtained by subjecting a sample of the composition to HSQC NMR, wherein the sample is dissolved in D 2 O.