Claims
- 1. A method to prepare a neurotrophic composition, which method comprises:
- (a) extracting factors from caudate-putamen tissue of a normal mammal;
- (b) assaying the caudate-putamen extract for trophic effects on dopaminergic neurons obtained from substantia nigra as measured by the ability to stimulate dopamine uptake by said neurons; and
- (c) isolating factors having said trophic effects.
- 2. The method of claim 1 wherein said normal mammal is of bovine source.
- 3. The method of claim 1 wherein said extraction comprises homogenizing the tissue extract in aqueous solution, centrifuging the homogenate, and recovering the supernatant.
- 4. The method of claim 1 which further comprises purifying the isolated factors by fractionation.
- 5. The method of claim 4 wherein said purification yields dopaminergic neurotrophic factors having molecular weights less than 3,000 daltons.
- 6. The method of claim 5 wherein a dopaminergic neurotrophic factor has a molecular weight range of about 1,000 to about 1,600 daltons.
- 7. The method of claim 5 wherein a dopaminergic neurotrophic factor has a molecular weight range of about 1,500 to about 1,800 daltons.
- 8. The method of claim 5 which further comprises purifying the dopaminergic neurotrophic factor by cation-exchange HPLC chromatography.
- 9. The method of claim 3 wherein said tissue extract is prepared from fresh tissue and said aqueous solution comprises a buffer supplemented with one or more protease inhibitors.
- 10. The method of claim 9 which further comprises purifying said isolated factors by gel filtration chromatography.
- 11. The method of claim 10 wherein said purification yields dopaminergic neurotrophic factors having molecular weights less than 3,000 daltons.
- 12. The method of claim 5 wherein a dopaminergic neurotrophic factor has a molecular weight range of about 1,800 to about 2,600 daltons.
- 13. The method of claim 11 wherein a dopaminergic neurotrophic factor has a molecular weight range of about 1,500 to about 1,800 daltons.
- 14. The method of claim 11 wherein a dopaminergic neurotrophic factor has a molecular weight range of about 1,000 to about 1,600 daltons.
- 15. The method of claim 11 wherein said purification further comprises purifying the dopaminergic neurotrophic factors by anion-exchange HPLC chromatography.
- 16. A factor prepared by the method of claim 7.
- 17. A factor prepared by the method of claim 12.
CROSS-REFERENCE TO RELATED APPLICATIONS
The present application is a continuation-in-part of U.S. Ser. No. 768,887, filed 23 Aug. 1985, now abandoned, which is a divisional of U.S. Ser. No. 444,293, filed 24 Nov. 1982, now abandoned.
US Referenced Citations (1)
Number |
Name |
Date |
Kind |
3989819 |
Brockman |
Nov 1976 |
|
Foreign Referenced Citations (3)
Number |
Date |
Country |
0082612 |
Nov 1982 |
EPX |
2420544 |
Nov 1979 |
FRX |
73526 |
Apr 1984 |
JPX |
Non-Patent Literature Citations (4)
Entry |
Appel, Ann Neurol, 1981, 10:499-505. |
Tomozawa and Appel, Society for Neuroscience Abstracts, No. 313.10, vol. II, Part 1; 15th Annual Meeting; Dallas, Texas 10/20/85. |
Appel et al; Tropic Factors and Degenerative Neuroligic Disease, May 19-20, 1986, International Conference on Neuroplasticity, Madrid, Spain. |
Tomozawa and Appel, Brain Research, 1986, 399:111-124. |
Divisions (1)
|
Number |
Date |
Country |
Parent |
444293 |
Nov 1982 |
|
Continuation in Parts (1)
|
Number |
Date |
Country |
Parent |
768887 |
Aug 1985 |
|