Methods of determining efficacy of cyclodextrin therapy

1. A method for determining efficacy of a cyclodextrin therapy in a subject afflicted with a disorder involving the accumulation of one or more oxysterols, comprising:

(a) obtaining at least one first body fluid sample from the subject prior to administering a cyclodextrin to the subject;

(b) administering the cyclodextrin to the subject;

(c) obtaining at least one second body fluid sample from the subject after the administering the cyclodextrin;

(d) subjecting the at least one first body fluid sample to a first chromatography-mass spectroscopy analysis to determine concentration of 24-hydroxycholesterol;

(e) subjecting the at least one second body fluid sample to a second chromatography-mass spectroscopy analysis to determine concentration of 24-hydroxycholesterol; and

(f) determining magnitude of difference between the 24-hydroxycholesterol concentration of the at least one first body fluid sample and the at least one second body fluid sample, whereby an increase or stabilization of 24-hydroxycholesterol concentration in the second sample compared to the first sample indicates efficacy of the cyclodextrin therapy.

2. A method in accordance with claim 1 , wherein each of the first chromatography-mass spectroscopy analysis and the second chromatography-mass spectroscopy analysis comprises:

(a) adding to the body fluid sample a known amount of an internal standard comprising an oxysterol other than 24(S)-hydroxycholesterol;

(b) isolating 24(S)-hydroxycholesterol from the biological sample; and

(c) quantifying the isolated 24(S)-hydroxycholesterol.

3. A method in accordance with claim 2 , wherein the quantifying 24(S)-hydroxycholesterol comprises determining the relative concentration of 24(S)-hydroxycholesterol and the known amount of internal standard in the body fluid sample by correlating the area under the curve obtained for the 24(S)-hydroxycholesterol with the area under the curve obtained for the internal standard.

4. A method in accordance with claim 1 , wherein the first biological sample and the second biological are each selected from the group consisting of plasma, serum, blood, sputum and amniotic fluid.

5. A method in accordance with claim 1 , wherein the body fluid sample is selected from the group consisting of plasma, serum and blood.

6. A method in accordance with claim 1 , wherein each of the first chromatography-mass spectroscopy analysis and the second chromatography-mass spectroscopy analysis is a liquid chromatography-mass spectroscopy analysis.

7. A method in accordance with claim 1 , wherein the disorder involving the accumulation of an oxysterol is selected from the group consisting of a lysosomal storage disease, a cholesterol trafficking disease, a neurodegenerative disease and a combination thereof.

8. A method in accordance with claim 1 , wherein the disorder involving the accumulation of an oxysterol is selected from the group consisting of Niemann-Pick C (NPC) disease, infantile neuronal ceroid lipofuscinosis, GM1 gangliosidosis (“GM-1”), GM-2 gangliosidosis (“GM-2”), Gaucher's disease and hepatosplenomegaly.

9. A method in accordance with claim 8 , wherein the GM-2 gangliosidosis (“GM-2”) is Tay-Sachs disease.

10. A method in accordance with claim 1 , wherein the disorder involving the accumulation of an oxysterol is Niemann-Pick C (NPC) disease.

11. A method for determining efficacy of a cyclodextrin therapy in a subject afflicted with a disorder involving the accumulation of one or more oxysterols, comprising:

(a) obtaining at least one first body fluid sample from the subject prior to administering a cyclodextrin to the subject;

(b) administering the cyclodextrin to the subject;

(c) obtaining at least one second body fluid sample from the subject after the administering the cyclodextrin;

(d) subjecting the at least one first body fluid sample to a first chromatography-mass spectroscopy analysis to determine concentration of cholestane-3β,5α,6β-triol;

(e) subjecting the at least one second body fluid sample to a second chromatography-mass spectroscopy analysis to determine concentration of cholestane-3β,5α,6β-triol; and

(f) determining magnitude of difference between the cholestane-3β,5α,6β-triol concentration of the at least one first body fluid sample and the at least one second body fluid sample, whereby a reduction in cholestane-3β,5α,6β-triol concentration in the second sample compared to the first sample indicates efficacy of the cyclodextrin therapy.

12. A method in accordance with claim 11 , wherein each of the first chromatography-mass spectroscopy analysis and the second chromatography-mass spectroscopy analysis comprises:

(a) adding to the body fluid sample a known amount of an internal standard comprising an oxysterol other than cholestane-3β,5α,6β-triol;

(b) isolating cholestane-3β,5α,6β-triol from the biological sample; and

(c) quantifying the isolated cholestane-3β,5α,6β-triol.

13. A method in accordance with claim 12 , wherein the quantifying cholestane-3β,5α,6β-triol comprises determining the relative concentration of cholestane-3β,5α,6β-triol and the known amount of internal standard in the body fluid sample by correlating the area under the curve obtained for the cholestane-3β,5α,6β-triol with the area under the curve obtained for the internal standard.

14. A method in accordance with claim 11 , wherein the first biological sample and the second biological are each selected from the group consisting of plasma, serum, blood, sputum and amniotic fluid.

15. A method in accordance with claim 11 , wherein the body fluid sample is selected from the group consisting of plasma, serum and blood.

16. A method in accordance with claim 11 , wherein each of the first chromatography-mass spectroscopy analysis and the second chromatography-mass spectroscopy analysis is a liquid chromatography-mass spectroscopy analysis.

17. A method in accordance with claim 11 , wherein the disorder involving the accumulation of an oxysterol is selected from the group consisting of a lysosomal storage disease, a cholesterol trafficking disease, a neurodegenerative disease and a combination thereof.

18. A method in accordance with claim 11 , wherein the disorder involving the accumulation of an oxysterol is selected from the group consisting of Niemann-Pick C (NPC) disease, infantile neuronal ceroid lipofuscinosis, GM1 gangliosidosis (“GM-1”), GM-2 gangliosidosis (“GM-2”), Gaucher's disease and hepatosplenomegaly.

19. A method in accordance with claim 18 , wherein the GM-2 gangliosidosis (“GM-2”) is Tay-Sachs disease.

20. A method in accordance with claim 11 , wherein the disorder involving the accumulation of an oxysterol is Niemann-Pick C (NPC) disease.