Treatment of alpha-galactosidase A deficiency

Abstract

The invention provides methods of treating α-galactosidase A deficiency. Dosage forms, methods of administration, and methods of analyzing human α-galactosidase A are also included.

We claim:  
     
         1 . A pharmaceutical composition comprising a human α-galactosidase A (α-Gal A) preparation, wherein the serum clearance of the α-Gal A preparation from human circulation is less than 4 ml/min/kg on the linear portion of the AUC vs. dose curve.  
     
     
         2 . The pharmaceutical composition of  claim 1 , wherein serum clearance of the α-Gal A preparation from human circulation is 3.5 mL/min/kg or less on the linear portion of the AUC vs. dose curve.  
     
     
         3 . The pharmaceutical composition of  claim 1 , wherein serum clearance of the α-Gal A preparation from human circulation is 3.0 mL/min/kg or less on the linear portion of the AUC vs. dose curve.  
     
     
         4 . The pharmaceutical composition of  claim 1 , wherein serum clearance of the α-Gal A preparation from human circulation is 2.5 mL/min/kg or less on the linear portion of the AUC vs. dose curve.  
     
     
         5 . A kit for the treatment of α-Gal A deficiency, comprising: (a) a human α-Gal A glycoprotein preparation; and (b) instructions to administer the preparation to a subject in need thereof, wherein the serum clearance of the α-Gal A preparation from subject's circulation is less than 4 mL/min/kg on the linear portion of the AUC vs. dose curve.  
     
     
         6 . The kit of  claim 5 , wherein the serum clearance of the α-Gal A preparation from subject's circulation is 3.5 mL/min/kg or less on the linear portion of the AUC vs. dose curve.  
     
     
         7 . The kit of  claim 5 , wherein the serum clearance of the α-Gal A preparation from subject's circulation is 3.0 mL/min/kg or less on the linear portion of the AUC vs. dose curve.  
     
     
         8 . The kit of  claim 5 , wherein the serum clearance of the α-Gal A preparation from subject's circulation is 2.5 mL/min/kg or less on the linear portion of the AUC vs. dose curve.  
     
     
         9 . The kit of  claim 5 ,  6 ,  7  or  8 , further comprising instructions to administer a unit dose of the α-Gal A preparation of between 0.05 mg and 2.0 mg per kilogram of body weight.  
     
     
         10 . The kit of  claim 9 , wherein the unit dose of the α-Gal A preparation is between 0.05 mg and 1.0 mg per kilogram of body weight.  
     
     
         11 . The kit of  claim 9 , wherein the unit dose of the α-Gal A preparation is between 0.05 mg and 0.5 mg per kilogram of body weight.  
     
     
         12 . The kit of  claim 9 , wherein the unit dose of the α-Gal A preparation is between 0.05 mg and less than 0.3 mg per kilogram of body weight.  
     
     
         13 . The kit of  claim 9 , further comprising instructions to administer the unit dose no more than once every 8 weeks.  
     
     
         14 . The kit of  claim 9 , further comprising instructions to administer the unit dose no more than once every 6 weeks.  
     
     
         15 . The kit of  claim 9 , further comprising instructions to administer the unit dose no more than once every 4 weeks.  
     
     
         16 . The kit of  claim 9 , further comprising instructions to administer the unit dose no more than once every 21 days.  
     
     
         17 . The kit of  claim 9 , further comprising instructions to administer the unit dose no more than once every 14 days.  
     
     
         18 . The kit of  claim 9 , further comprising instructions to administer the unit close no more than once every 10 days.  
     
     
         19 . A kit for the treatment of α-Gal A deficiency, comprising: a human α-Gal A glycoprotein preparation and instructions to administer the preparation to a subject in need thereof at a unit dose of less than 1.0 mg per kilogram of body weight of the subject.  
     
     
         20 . The kit of  claim 19 , wherein the unit dose is less than 0.5 mg per kilogram of body weight of the subject.  
     
     
         21 . The kit of  claim 19 , wherein the unit dose is less than 0.3 mg per kilogram of body weight of the subject.  
     
     
         22 . The kit of  claim 19 ,  20  or  21 , further comprising instructions to administer the unit dose no more than once every 8 weeks.  
     
     
         23 . The kit of  claim 22 , wherein the kit comprises instructions to administer the unit dose no more than once every 6 weeks.  
     
     
         24 . The kit of  claim 22 , wherein the kit comprises instructions to administer the unit dose no more than once every 4 weeks.  
     
     
         25 . The kit of  claim 22 , wherein the kit comprises instructions to administer the unit dose no more than once every 21 days.  
     
     
         26 . The kit of  claim 22 , wherein the kit comprises instructions to administer the unit dose no more than once every 14 days.  
     
     
         27 . The kit of  claim 22 , wherein the kit comprises instructions to administer the unit dose no more than once every 10 days.  
     
     
         28 . A kit for the treatment of α-Gal A deficiency, comprising a human α-Gal A glycoprotein preparation and instructions to administer the preparation to a subject in need thereof no more than once every 8 weeks.  
     
     
         29 . The kit of  claim 28 , wherein the instructions comprise instructions to administer the preparation to a subject in need thereof no more than once every 6 weeks.  
     
     
         30 . The kit of  claim 28 , wherein the instructions comprise instructions to administer the preparation to a subject in need thereof no more than once every 4 weeks.  
     
     
         31 . The kit of  claim 28 , wherein the instructions comprise instructions to administer the preparation to a subject in need thereof no more than once every 21 days.  
     
     
         32 . The kit of  claim 28 , wherein the instructions comprise instructions to administer The preparation to a subject in need thereof no more than once every 14 days.  
     
     
         33 . The kit of  claim 28 , wherein the instructions comprise instructions to administer the preparation to a subject in need thereof no more than once every 10 days.  
     
     
         34 . The kit of any one of claims  28 - 33 , further comprising instructions to administer a unit dose of the α-Gal A preparation of between 0.05 mg and 2.0 mg per kilogram of body weight.  
     
     
         35 . The kit of  claim 34 , wherein the unit dose is between 0.05 and 1.0 mg per kilogram of body weight.  
     
     
         36 . The kit of  claim 34 , wherein the unit dose is between 0.05 and 0.5 mg per kilogram of body weight.  
     
     
         37 . The kit of  claim 34 , wherein the unit dose is between 0.05 and less than 3 mg per kilogram of body weight.  
     
     
         38 . A method of selecting a unit dose range of α-Gal A for treatment of a subject having an α-Gal A deficiency, the method comprising: providing the body weight of the subject, and determining the range between 0.05 mg and 1 mg of α-Gal A per kilogram of body weight of the subject, thereby selecting a unit dose range.  
     
     
         39 . The method of  claim 38 , the range determined is between 0.05 mg and 0.5 mg of α-Gal A per kilogram of body weight of the subject.  
     
     
         40 . The method of  claim 38 , the range determined is between 0.05 mg and less than 0.3 mg of α-Gal A per kilogram of body weight of the subject.  
     
     
         41 . A method of treating a subject, comprising administering to a subject in need thereof a human α-gal A glycoprotein preparation, wherein the serum clearance of the α-Gal A preparation from the subject's circulation is less than 4 mL/min/kg on the lineal portion of the AUC vs. dose curve.  
     
     
         42 . The method of  claim 41 , wherein serum clearance of the α-Gal A preparation from subject's circulation is 3.5 mL/min/kg or less on the linear portion of the AUC vs. dose curve.  
     
     
         43 . The method of  claim 41 , wherein serum clearance of the α-Gal A preparation from human circulation is 3.0 mL/min/kg or less on the linear portion of the AUC vs. dose curve.  
     
     
         44 . The method of  claim 1 , wherein serum clearance of the α-Gal A preparation from human circulation is 2.5 mL/min/kg or less on the linear portion of the AUC vs. dose curve.  
     
     
         45 . A method of treating a subject, comprising administering to a subject in need thereof a human α-gal A glycoprotein preparation at a unit dose of less than 1.0 mg per kilogram of body weight of the subject.  
     
     
         46 . The method of  claim 45 , wherein the unit dose is less than 0.5 mg per kilogram of body weight of the subject.  
     
     
         47 . The method of  claim 45 , wherein the unit dose is less than 0.3 mg per kilogram of body weight of the subject.  
     
     
         48 . The method of  claim 45 , wherein the unit dose is less than 0.25 mg per kilogram of body weight of the subject.  
     
     
         49 . The method of  claim 45 , wherein the unit dose is less than 0.2 mg per kilogram of body weight of the subject.  
     
     
         50 . The method of any one claims  45 - 49 , wherein the unit dose is administered no more than once every 8 weeks.  
     
     
         51 . The method of  claim 50 , wherein the unit dose is administered no more than once every 6 weeks.  
     
     
         52 . The method of  claim 50 , wherein the unit dose is administered no more than once every 4 weeks.  
     
     
         53 . The method of  claim 50 , wherein the unit dose is administered no more than once every 21 days.  
     
     
         54 . The method of  claim 50 , wherein the unit dose is administered no more than once every 14 days.  
     
     
         55 . The method of  claim 50 , wherein the preparation is administered for at least 1 year.  
     
     
         56 . The method of  claim 51 , wherein the preparation is administered for at least 1 year.  
     
     
         57 . The method of  claim 52 , wherein the preparation is administered for at least 1 year.  
     
     
         58 . The method of  claim 53 , wherein the preparation is administered for at least 1 year.  
     
     
         59 . A method of treating a subject, comprising administering to a subject in need thereof a human (α-Gal A glycoprotein preparation at least twice but no more than once every 14 days.  
     
     
         60 . The method of  claim 59 , wherein the preparation is administered no more than once every 4 weeks.  
     
     
         61 . The method of  claim 59 , wherein the preparation is administered no more than once every 6 weeks.  
     
     
         62 . The method of  claim 59 , wherein the preparation is administered no more than once every 8 weeks.  
     
     
         63 . The method of  claim 59 ,  60 ,  61  or  62 , wherein the unit close of the administration is less than 0.5 mg per kilogram of body weight.  
     
     
         64 . The method of  claim 63 , wherein the unit dose of the administration is less than 0.3 mg per kilogram of body weight.  
     
     
         65 . The method of  claim 45 ,  46 ,  47  or  48 , wherein the unit dose saturates liver uptake of the α-Gal A.  
     
     
         66 . A method for analyzing an α-Gal A preparation, the method comprising: 
 obtaining or providing a first test α-Gal A preparation; and  
 determining if the first test α-Gal A preparation has one or more of the characteristics (1)-(7): 
 (1) has at least about 75% neutral, mono- and di-sialylated glycans combined;  
 (2) has less than about 35% tri- and tetra-sialylated glycans combined;  
 (3) has greater than 50% complex glycans;  
 (4) has less than about 45% phosphorylated glycans;  
 (5) has greater than about 45% sialylated glycans;  
 (6) has a ratio of sialic acid to mannose-6-phosphate on a mole per mole basis greater than 1.5 to 1; and  
 (7) has a ratio of sialylated glycans to phosphorylated glycans greater than 1, thereby analyzing an α-Gal A preparation.  
 
 
     
     
         67 . The method of  claim 66 , further comprising the step of selecting the test α-Gal A preparation if it has one or more of the characteristics (1)-(7).  
     
     
         68 . The method of  claim 66 , wherein the determining step comprises determining if the test α-Gal A preparation has at least about 75% neutral, mono- and di-sialylated glycans combined.  
     
     
         69 . The method of  claim 66 , wherein the determining step comprises determining if the test α-Gal A preparation has less than about 35% tri- and tetra-sialylated glycans combined.  
     
     
         70 . The method of  claim 66 , wherein the determining step comprises determining if the test α-Gal A preparation has greater than 50% complex glycans.  
     
     
         71 . The method of  claim 66 , wherein the determining step comprises determining if the test α-Gal A preparation has less than about 45% phosphorylated glycans.  
     
     
         72 . The method of  claim 66 , wherein the determining step comprises determining if the test α-Gal A preparation has greater than about 45% sialylated glycans.  
     
     
         73 . The method of  claim 66 , wherein the determining step comprises determining if the test α-Gal A preparation has a ratio of sialic acid to mannose-6-phosphate on a mole per mole basis greater than 1.5 to 1.  
     
     
         74 . The method of  claim 66 , wherein the determining step comprises determining if the test α-Gal A preparation has a ratio of sialylated glycans to phosphorylated glycans greater than 1.  
     
     
         75 . The method of  claim 66 , wherein the method further comprises the step of entering the result of the determination into a record.  
     
     
         76 . The method of  claim 66 , wherein the method comprises determining if the test α-Gal A preparation has two or more of the characteristics (1)-(7).  
     
     
         77 . The method of  claim 66 , wherein the determination is performed by one or more methods chosen from the group consisting: of ion exchange chromatography, high performance anion exchange (HPAE) chromatography, high performance liquid chromatography (HPLC), mass spectroscopy.  
     
     
         78 . The method of  claim 66 , wherein the α-Gal A sample is harvested from a mammalian cell.  
     
     
         79 . The method of  claim 78 , wherein the mammalian cell is a human cell.  
     
     
         80 . The method of  claim 78 , wherein the mammalian cell is a non-human cell.  
     
     
         81 . The method of  claim 78 , wherein the mammalian cell is a CHO cell.  
     
     
         82 . The method of  claim 66 , wherein a carbohydrate signature of the test preparation has been modified before the determining step is performed.  
     
     
         83 . The method of  claim 82 , wherein a carbohydrate signature of the test preparation has been modified by treatment with an enzyme.  
     
     
         84 . The method of  claim 83 , wherein the enzyme is a glycosidase, glycosyl transferase, phosphoryl transferase, kinase or sialyl transferase.  
     
     
         85 . The method of  claim 82 , wherein the carbohydrate signature of the test preparation has been modified by treatment with a phosphatase inhibitor.  
     
     
         86 . The method of  claim 82 , wherein the carbohydrate signature of the test preparation has been modified by glyco-engineering.  
     
     
         87 . The method of  claim 82 , wherein the carbohydrate signature of the test preparation has been modified by treatment with an inhibitor of glycosylation.  
     
     
         88 . The method of  claim 66 , further comprising the step of comparing the test human α-Gal A preparation to a reference α-Gal A preparation.  
     
     
         89 . The method of  claim 88 , wherein the reference α-Gal A preparation is a human α-Gal A preparation made in human cells.  
     
     
         90 . The method of  claim 66 , further comprising the steps of: 
 obtaining or providing a second test α-Gal A preparation;    determining if the second preparation has one or more of the characteristics (1)-(7); and    entering the result of each determination into a record,    wherein the first and second preparations are first and second batches of a pharmaceutical α-Gal A preparation.    
     
     
         91 . The method of  claim 66 , further comprising the step of: 
 predicting a pharmacokinetic parameter or biological activity of the test α-Gal A preparation, wherein the pharmacokinetic parameter or biological activity is predicted to be desirable if the test α-Gal A preparation has one or more of the characteristics (1)-(7).    
     
     
         92 . The method of  claim 66  further comprising the steps of: 
 evaluating a pharmacokinetic parameter or biological activity of the test α-Gal A preparation.  
 
     
     
         93 . The method of  claim 91  or  92 , wherein the pharmacokinetic parameter or biological activity is selected from the group consisting of: enzymatic activity, serum clearance and tissue uptake.  
     
     
         94 . The method of  claim 91  or  92 , wherein the pharmacokinetic parameter or biological activity is selected from the group consisting of: liver uptake, renal uptake and cardiovascular uptake.  
     
     
         95 . The method of  claim 91  or  92 , wherein the pharmacokinetic parameter or biological activity is tissue targeting to at least one of: liver endothelial cells, liver sinusoidal cells, capillary/vascular endothelial cells, renal glomerular epithelial cells (podocytes), glomerular mesangial cells, renal endothelial cells, pulmonary cells, renal cells, neural cells, or cardiac myocytes.  
     
     
         96 . The method of  claim 91 , further comprising the step of using the prediction to design an α-Gal A therapeutic preparation for a specific patient or a specific variant of Fabry disease.  
     
     
         97 . The method of  claim 96 , wherein the specific variant of Fabry disease is renal variant Fabry disease or cardiac variant Fabry disease.  
     
     
         98 . A method for analyzing an α-Gal A preparation, the method comprising: 
 obtaining or providing a test α-Gal A preparation; and determining one or more of: (1) if the serum clearance from human circulation is less than 4 mL/min/kg on the linear portion of the AUC vs. dose curve; (2) if the preparation is preferentially targeted to capillary/vascular endothelial cells, renal glomerular epithelial cells (podocytes), glomerular mesangial cells, renal endothelial cells, pulmonary cells, renal cells, neural cells, or cardiac myocytesin a subject, and (3) is not taken up by liver hepatocytes, thereby analyzing an α-Gal A preparation.  
 
     
     
         99 . The method of  claim 98 , wherein the method further comprises the step of entering the result of the determination into a record.  
     
     
         100 . A method of producing an improved human α-Gal A preparation, the method comprising the steps of: 
 a. providing a human (α-Gal A preparation harvested from a cell; and  
 b. modifying the carbohydrate signature of the α-Gal A preparation to match one or more of the following parameters: 
 (1) has at least about 75% neutral, mono- and di-sialylated glycans combined;  
 (2) has less than about 35% tri- and tetra-sialylated glycans combined;  
 (3) has greater than 50% complex glycans;  
 (4) has less than about 45% phosphorylated glycans;  
 (5) has greater than about 45% sialylated glycans;  
 (6) has a ratio of sialic acid to mannose-6-phosphate on a mole per mole basis greater than 1.5 to 1; and  
 (7) has a ratio of sialylated glycans to phosphorylated glycans greater than 1, thereby providing an improved α-Gal A preparation.  
 
 
     
     
         101 . The method of  claim 100 , wherein the carbohydrate signature of the α-Gal A preparation is modified by glycoengineering.  
     
     
         102 . The method of  claim 101 , wherein the carbohydrate signature of the α-Gal A preparation is modified by one or both of: genetically engineering the cell to produce a human α-Gal A having a non-naturally occurring glycosylation site; and genetically engineering the cell to produce a glucosidase, glycosyl transferase, phosphoryl transferase, phosphatase, or sialyl transferase.  
     
     
         103 . The method of  claim 100 , wherein the carbohydrate signature of the α-Gal A preparation is modified by one or more of: selective isolation of glycoforms during the α-Gal A purification process; treatment of the cell or preparation with a carbohydrate modifying enzyme; and treatment of the cell or preparation with an inhibitor of glycosylation.  
     
     
         104 . The method of  claim 100 , further comprising the step of analyzing the carbohydrate signature of the α-Gal A preparation after modification.  
     
     
         105 . A method of treating a subject, comprising: 
 providing or obtaining a panel of two or more α-Gal A preparations having different carbohydrate signatures;    selecting an α-Gal A preparation having a carbohydrate signature that matches one or more of he following parameters: 
 (1) has at least about 75% neutral, mono- and di-sialylated glycans combined;  
 (2) has less than about 35% tri- and tetra-sialylated glycans combined;  
 (3) has greater than 50% complex glycans;  
 (4) has less than about 45% phosphorylated glycans;  
 (5) has greater than about 45% sialylated glycans;  
 (6) has a ratio of sialic acid to mannose-6-phosphate on a mole per mole basis greater than 1.5 to 1; and  
 (7) has a ratio of sialylated glycans to phosphorylated glycans greater than 1; and  
   administering one or more doses of a therapeutically effective amount of the selected preparation to the subject.    
     
     
         106 . The method of  claim 105 , further comprising evaluating the tissue distribution or serum clearance of the α-Gal A preparation in the subject.  
     
     
         107 . The method of  claim 106 , wherein the evaluating step is performed repeatedly over time.  
     
     
         108 . The method of  claim 106 , further comprising adjusting the dose of the α-Gal A preparation after the evaluation step.  
     
     
         109 . The method of  claim 105 , wherein the method further comprises monitoring the status of the subject in response to the administration of the α-Gal A preparation.  
     
     
         110 . A method of selecting a batch of an α-Gal A preparation, the method comprising: 
 providing a plurality of batches of α-Gal A, each of the plurality having a batch-to-batch variation in carbohydrate signature; and  
 selecting a batch with less than a preselected range of variation from one or more of the following parameters in carbohydrate signature: 
 (1) has at least about 75% neutral, mono- and di-sialylated glycans combined;  
 (2) has less than about 35% tri- and tetra-sialylated glycans combined;  
 (3) has greater than 50% complex glycans;  
 (4) has less than about 45% phosphorylated glycans;  
 (5) has greater than about 45% sialylated glycans;  
 (6) has a ratio of sialic acid to mannose-6-phosphate on a mole per mole basis greater than 1.5 to 1; and  
 (7) has a ratio of sialylated glycans to phosphorylated glycans greater than 1.  
 
 
     
     
         111 . The method of  claim 110 , wherein the preselected variation is less than 5%.  
     
     
         112 . The method of  claim 110 , wherein the preselected variation is less than 2.5%.