Oligosaccharide compositions and methods of use thereof for reducing ammonia levels
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 hyperammonemia-related diseases (e.g., urea cycle disorders and hepatic encephalopathy).
1. An oligosaccharide composition comprising a plurality of oligosaccharides, the plurality of oligosaccharides being characterized by a multiplicity-edited gradient-enhanced 1 H- 13 C heteronuclear single quantum coherence (HSQC) NMR spectrum comprising signals 1, 3, and 4, each signal having properties as follows:
Area under the curve (AUC)
Center Position
(% of total areas of signals observed
(ppm)
and assigned to the oligosaccharide
Glycosidic
Signal
1 H
13 C
composition)
Bond Type
1
5.39
100.31
3.37-4.78
alpha-1,4
3
5.35
99.77
6.64-8.71
alpha-1,3
4
5.22
102.55
0.00-0.95
—.
2. The composition of claim 1 , wherein the spectrum further comprises 1-2 signals selected from signals 8 and 9, each signal having properties as follows:
Area under the curve (AUC)
Center Position
(% of total areas of signals observed
(ppm)
and assigned to the oligosaccharide
Glycosidic
Signal
1 H
13 C
composition)
Bond Type
8
5.03
93.95
0.00-0.89
—
9
4.97
98.49
32.69-40.14
alpha-1,6.
3. The composition of claim 1 , wherein the spectrum further comprises 1-6 signals selected from signals 6, 7, 11, 12, 13, and 14, each signal having properties as follows:
Area under
the curve (AUC)
(% of total areas of
signals observed
Center Position
and assigned
(ppm)
to the oligosaccharide
Glycosidic
Signal
1 H
13 C
composition)
Bond Type
6
5.17
96.06
0.26-1.55
—
7
5.1
96.77
0.65-2.30
alpha-1,2-alpha
11
4.71
103.64
6.78-8.16
beta-1,3
12
4.64
103.54
1.23-3.20
beta-1-alpha-1′
13
4.63
104.53
1.80-3.09
beta-1,2-alpha
14
4.52
103.34
28.46-32.06
beta-1,4/beta-1,6 co-
peak.
4. The composition of claim 1 , wherein the spectrum further comprises 1-3 signals selected from signals 2, 5, and 10, each signal having properties as follows:
Area under
the curve (AUC)
(% of total areas of
signals observed
Center Position
and assigned
Glycosidic
(ppm)
to the oligosaccharide
Bond
Signal
1 H
13 C
composition)
Type
2
5.36
98.48
0.69-1.78
alpha-1,2-beta
5
5.19
93.95
1.56-4.45
alpha-1-alpha-1′
10
4.8
96.77
0.26-1.35
beta-1,2-beta,
reducing.
5. The composition of claim 1 , wherein the composition comprises signals 1-14, wherein each is 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
5.39
5.4229
5.3620
100.31
99.9518
100.6747
2
5.36
5.3877
5.3310
98.48
98.2048
98.7590
3
5.35
5.3971
5.2958
99.77
99.3614
100.1807
4
5.22
5.2357
5.1994
102.55
102.1205
102.9759
5
5.19
5.2047
5.1684
93.95
93.6506
94.2530
6
5.17
5.1933
5.1540
96.06
95.7590
96.3614
7
5.1
5.1207
5.0772
96.77
96.5181
97.0120
8
5.03
5.0545
5.0009
109.19
108.7831
109.6024
9
4.97
5.0194
4.9173
98.49
97.8795
99.0964
10
4.8
4.8197
4.7827
96.77
96.5904
96.9518
11
4.71
4.7641
4.6658
103.64
103.2169
104.0723
12
4.64
4.6670
4.6193
103.54
103.1807
103.9036
13
4.63
4.6514
4.6008
104.53
104.2289
104.8313
14
4.52
4.5841
4.4476
103.34
102.8193
103.8675.
6. The 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. 6 , acquisition parameters and processing parameters:
Acquisition parameters
1 H Carrier Frequency=4 ppm
13 C Carrier Frequency=65 ppm
Number of points in acquisition dimension=596
Spectral range in acquisition dimension=6.00 ppm to 2.03 ppm
Number of points in indirect dimension=300 complex points
Spectral range in indirect dimension=120 ppm to 10 ppm
Recycle delay=1 second
One-bond 1 H- 13 C coupling constant=J CH =146 Hz
Number of scans=8
Temperature=298 K
Solvent=D2O
Processing parameters
Window function in direct dimension=Gaussian broadening, 7.66 Hz
Window function in indirect dimension=Gaussian broadening 26.48 Hz
Processing=512 complex points in direct dimension, 1024 complex points in indirect dimension.
7. The composition of claim 1 , wherein the mean degree of polymerization (DP) of the oligosaccharide composition is from about DP5 to about DP11.
8. A method of reducing ammonia levels in a human subject, comprising administering to the human subject an effective amount of an oligosaccharide composition according to claim 1 .
9. A method of reducing the rate of hyperammonemic crises in a human subject having or having been diagnosed with a disease or condition associated with hyperammonemia, comprising administering to the human subject an effective amount of an oligosaccharide composition according to claim 1 .
10. The method of claim 9 , wherein the subject has or has been diagnosed as having a urea cycle disorder or hepatic encephalopathy (HE).
11. A method for treating encephalopathy in a human subject, comprising administering to the human subject an effective amount of an oligosaccharide composition according to claim 1 .
12. The composition of claim 1 , wherein the composition comprises 16% to 24% dextrose equivalent (dry basis).
13. An oligosaccharide composition comprising a plurality of oligosaccharides, each oligosaccharide comprising a plurality of monomer radicals;
the plurality of oligosaccharides comprising two or more types of monomer radicals selected from radicals (1)-(7), wherein the selected radicals are present in the plurality of oligosaccharides in a defined molar percentage (mol %) range:
(1) 2-glucopyranose diradicals, representing 6.15-9.54 mol % of monomer radicals in the plurality of oligosaccharides;
(2) 6-glucopyranose diradicals, representing at least 18.94 mol % of monomer radicals in the plurality of oligosaccharides;
(3) 4-glucopyranose diradicals, representing 6.63-8.74 mol % of monomer radicals in the plurality of oligosaccharides;
(4) 3,4-glucopyranose triradicals, representing less than 1.42 mol % of monomer radicals in the plurality of oligosaccharides;
(5) 2,3-glucopyranose triradicals, representing 1.01-2.59 mol % of monomer radicals in the plurality of oligosaccharides;
(6) 3,4,6-glucopyranose tetraradicals, representing less than 1.00 mol % of monomer radicals in the plurality of oligosaccharides; and
(7) 2,4,6-glucopyranose tetraradicals, representing less than 0.72 mol % of monomer radicals in the plurality of oligosaccharides;
wherein at least one of the two or more types of monomer radicals is (4) or (7).
14. The composition of claim 13 , further comprising one or more types of monomer radicals selected from radicals (8)-(12):
(8) t-glucopyranose monoradicals, representing 26.87-46.98 mol % of monomer radicals in the plurality of oligosaccharides;
(9) 3-glucopyranose diradicals, representing at least 6.94 mol % of monomer radicals in the plurality of oligosaccharides;
(10) 2,4-glucopyranose triradicals, representing less than 1.43 mol % of monomer radicals in the plurality of oligosaccharides;
(11) 2,6-glucopyranose/4,6-glucopyranose triradicals, representing less than 9.66 mol % of monomer radicals in the plurality of oligosaccharides; and
(12) 2,3,4-glucopyranose radicals, representing 0.07-0.85 mol % of monomer radicals in the plurality of oligosaccharides.
15. The composition of claim 13 , further comprising one or more types of monomer radicals selected from radicals (13)-(15):
(13) 3,6-glucopyranose triradicals, representing 2.35-6.01 mol % of monomer radicals in the plurality of oligosaccharides;
(14) 2,3,6-glucopyranose tetraradicals, representing 0.06-0.86 mol % of monomer radicals in the plurality of oligosaccharides; and
(15) 2,3,4,6-glucopyranose pentaradicals, representing less than 0.61 mol % of monomer radicals in the plurality of oligosaccharides.