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US 7,968,082 B1Granted
Evaluating mixtures of low molecular weight heparins by NMR
AssigneeMomenta Pharmaceuticals, Inc.
InventorsZachary Shriver, Daniela Beccati, Ishan Capila, Jonathan Lansing
Life of this patent
Estimating the term…
Abstract
Methods for analyzing mixtures of polysaccharides, for example heparin such as enoxaparin are described. In some instances, the mixtures are analyzed using 1D NMR and/or 2D NMR.
Claims21
A method of processing an enoxaparin preparation comprising:
- performing 2D-NMR to determine an amount of an epoxide, a reducing end 2-O sulfated uronic acid and a 1,6 anhydro mannosamine present in an enoxaparin preparation, and
- if the amount of the epoxide, the reducing end 2-O sulfated uronic acid and the 1,6 anhydro mannosamine is present in an amount within a preselected range, processing said preparation, wherein said processing includes one or more of selecting, accepting, processing into drug product, shipping, formulating, labeling, packaging or selling said preparation, and
- if the amount of the epoxide, the reducing end 2-O sulfated uronic acid or the 1,6 anhydro mannosamine is not present in an amount within the preselected range, discarding or withholding the preparation,
- wherein the preselected range of the epoxide corresponds to between 0.6 and 1.0 percent composition, wherein the preselected range of the reducing end 2-O sulfated uronic acid corresponds to between 0.8 and 1.4 percent composition, wherein the preselected range of the mannosamine corresponds to between 2.1 and 2.9 percent composition, and
- wherein the percent composition is calculated using the specific cross peak volumes for which all uronic acid residues have similar 1JCH couplings as do all glucosamine residues.
2 more elements
A method of processing an enoxaparin preparation comprising:
- performing 2D-NMR to determine a total amount of N-sulfated hexosamine (HNS), a total amount of 1,6 anhydrohexosamine, a total amount of 2-O-sulfated uronic acid, a total amount of 3-O-sulfated glucosamine (HNS,3S,6S), a total amount of N-acetyl glucosamine (NNAc), a total amount of 6-O-sulfated glucosamine (H6S), a total amount of non-sulfated uronic acid and a total amount of epoxide in an enoxaparin preparation,
- comparing the total amount of N-sulfated hexosamine (HNS), 1,6 anhydrohexosamine, 2-O-sulfated uronic acid, 3-O-sulfated glucosamine (HNS,3S,6S), N-acetyl glucosamine (HNAc), 6-O-sulfated glucosamine (H6S), non-sulfated uronic acid and epoxide to a preselected range, wherein the preselected range corresponds to a range A, range B or range C provided in Table 3,
- TABLE 3
- Percentage
- Percentage
- Percentage
- Composition
- Composition
- Composition
- Monosaccharide
- Range A*
- Range B
- Range C
- Hexosamine Residues
- Total N-sulfated
- 84-88
- 80-90
- 77-95
- hesoxamine (HNS)
- HNS-G
- 10.7-11.2
- 10.5-11.5
- 9.5-12.0
- HNS-I
- 7.0-8.0
- 6.7-8.0
- 6.3-8.2
- 1,6 An Man
- 2.3-2.7
- 2.2-2.8
- 2.1-2.9
- 1,6 An Glc
- 2.7-2.8
- 2.6-3.0
- 2.5-3.1
- Total 1,6 anhydro
- 5.2-5.4
- 5.0-5.6
- 4.6-5.8
- 3-O-sulfated
- 3.9-4.4
- 3.7-4.5
- 3.5-4.6
- glucosamine
- (HNS,3S,6S)
- N-acetyl
- 9.0-10.0
- 8.9-10.2
- 8.8-10.8
- glucosamine (HNAc)
- 6-O-sulfated
- 83-88
- 78-90
- 75-92
- glucosamine (H6S)
- Uronic acid residues
- 2-O-sulfated
- 74-76
- 71-80
- 67-83
- uronic acid
- I2S,red
- 0.9-1.2
- 0.9-1.3
- 0.8-1.4
- G2S
- 2.6-3.0
- 2.5-3.1
- 2.5-3.3
- Non-sulfated
- 24-26
- 23-27
- 22-28
- uronic acid
- 1-HNS/Ac,6S
- 5.3-6.4
- 5.0-6.5
- 4.7-6.5
- I-HNS,Ac
- 0.90-1.40
- 0.8-1.50
- 0.8-1.55
- G-HNS
- 12.5-13.5
- 11.8-13.7
- 11.3-13.9
- G-HNAc
- 3.10-4.40
- 3.00-4.60
- 3.00-4.85
- Other residues
- Epoxide
- 0.7-0.9
- 0.65-0.95
- 0.6-1.0
- wherein the percent composition is calculated using the anomeric cross peak volumes for which all uronic acid residues have similar 1JCH couplings as do all glucosamine residues;
- and,
- if the amounts are within the preselected range, processing said preparation, wherein processing includes one or more of selecting, accepting, processing into drug product, shipping, formulating, labeling, packaging or selling said preparation, and
- if the amounts are not within the preselected range, discarding or withholding the preparation.
96 more elements
Dependencies are inferred from the claim text.