EP0327769A2
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EP
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Novel neuronotrophic factor
Assignee: Fidia SpA
•
Filing: 1988-02-17
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Granted: —
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Expires: 2008-02-17
Abstract
Abstract A novel neuronotrophic factor having a molecular weight of about 14,000 to 17,000 daltons and an isoelectric point of about 10 is prepared by homogenization of mammalian brain tissue, particularly bovine brain tissue, acid precipitation of the homogenate thus produced, dialysis of the resulting supernatant with dialysis membranes having a molecular weight cut-off of between 5 and 10 kilodaltons and chromatographic fractionation upon molecular weight permeation of the dialyzed supernatant thus produced. The neuronotrophically active fractions may be further purified by cation exchange chromatography with a gradient of ammonium acetate buffer. The neuronotrophic factor of the invention is useful in the treatment of various neuropathological conditions.
Inventors
- 👤 Francesco Della Valle
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-08 03:23
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/435; C07K14/475; A; A61; A61K; A61K35/00; A61K35/12; A61K35/30; A61K38/00
Claims Analysis
Total Claims: 38
Claim 1
Independent Claim
82 words
A process for the preparation of a neuronotrophic factor which is a basic protein having a molecular weight of about 14,000 to 17,000 daltons characterized by the steps of (a) homogenization of mammalian brain tissue, (b) acid precipitation of the homogenate thus produced, (c) dialysis of the resulting supernatant with a dialysis membrane having a molecular weight restriction of between 5 and 10 kilodaltons and (d) chromatographic fractionation of the dialyzed supernatant to separate the supernatant components according to their molecular weight.
Claim 2
Dependent Claim
82 words
A process for the preparation of a neuronotrophic factor which is a basic protein having a molecular weight of about 14,000 to 17,000 daltons characterized by the steps of (a) homogenization of mammalian brain tissue, (b) acid precipitation of the homogenate thus produced, (c) dialysis of the resulting supernatant with a dialysis membrane having a molecular weight restriction of between 5 and 10 kilodaltons and (d) chromatographic fractionation of the dialyzed supernatant to separate the supernatant components according to their molecular weight.
Claim 3
Dependent Claim
31 words
A process according to claim 1, further comprising the step of (e) purification of the neuronotrophically active fractions thus obtained by cation exchange chromatography with a gradient of ammonium acetate buffer.
Claim 4
Dependent Claim
18 words
A process according to claim 1 or claim 2, wherein the mammalian brain tissue is bovine brain tissue.
Claim 5
Dependent Claim
21 words
A process according to claim 1 or claim 2, wherein caudate nuclei from bovine brains are subjected to said process steps.
Claim 6
Dependent Claim
19 words
A process according to claim 1, wherein step (b) is conducted at a pH of from 4 to 5.
Claim 7
Dependent Claim
32 words
A process according to claim 1, wherein the chromatographic fractionation step (d) is carried out on a molecular sieve with a diluted buffered eluent at a concentration of from 10nM to 30nM.
Claim 8
Dependent Claim
26 words
A process according to claim 6, wherein the fractionation is carried out using a stationary phase with a fractionation range of between 5,000 and 150,000 daltons.
Claim 9
Dependent Claim
42 words
A process according to claim 2, wherein the neuronotrophically active fractions are eluted from the cation exchange chromatography column with a gradient of ammonium acetate buffer having a concentration of from 0.1M to 1M and a pH of from 6 to 7.
Claim 10
Dependent Claim
23 words
A process according to claim 1 or claim 2, wherein the process steps are conducted at a temperature of from 0° to 6°C.
Claim 11
Dependent Claim
19 words
A process according to claim 1 or claim 2, wherein the neuronotrophically active fractions are pooled together and freeze-dried.
Claim 12
Dependent Claim
16 words
A neuronotrophic factor which is a basic protein having a molecular weight of about 14,400 daltons.
Claim 13
Dependent Claim
16 words
A neuronotrophic factor which is a basic protein having a molecular weight of about 14,400 daltons.
Claim 14
Dependent Claim
15 words
The neuronotrophic factor according to claim 11 which has an isoelectric point of about 10.
Claim 15
Dependent Claim
51 words
The neuronotrophic factor according to claim 11 which has a specific activity of at least about 0.01 mg/trophic units, in which the aforesaid trophic unit is the protein concentration (µg/ml) which favours survival of half the maximum number of neurons surviving in response to the saturating concentration of the active material.
Claim 16
Dependent Claim
28 words
A neuronotrophic factor which is a basic protein derived from the mammalian brain caudate nucleus, having neuronotrophic activity and a molecular weight of about 14,000 to 17,000 daltons.
Claim 17
Dependent Claim
28 words
A neuronotrophic factor which is a basic protein derived from the mammalian brain caudate nucleus, having neuronotrophic activity and a molecular weight of about 14,000 to 17,000 daltons.
Claim 18
Dependent Claim
15 words
The neuronotrophic factor according to claim 14 which has an isoelectric point of about 10.
Claim 19
Dependent Claim
11 words
A neuronotrophic factor prepared according to the process of claim 1.
Claim 20
Dependent Claim
33 words
A neuronotrophic factor according to claim 16 which is the fraction represented by the full horizontal bar in Figure 1 of the drawings when the said chromatographic fractionation is conducted with Sephadex G-150.
Claim 21
Dependent Claim
11 words
A neuronotrophic factor prepared according to the process of claim 2.
Claim 22
Dependent Claim
23 words
A neuronotrophic factor according to claim 18 which is the fraction represented by the full horizontal bar in Figure 2 of the drawings.
Claim 23
Dependent Claim
25 words
A neuronotrophic factor prepared according to the process of claim 1 or claim 2 which is capable of enhancing survival and differentiation of neuronal cells.
Claim 24
Dependent Claim
24 words
A pharmaceutical composition comprising a neuronotrophically active quantity of a neuronotrophic factor according to claim 11, and a pharmaceutically acceptable excipient or diluent vehicle.
Claim 25
Dependent Claim
27 words
A pharmaceutical composition comprising a neuronotrophically active quantity of a neuronotrophic factor according to claim 14 or claim 15, and a pharmaceutically acceptable excipient or diluent vehicle.
Claim 26
Dependent Claim
30 words
A pharmaceutical composition comprising a neuronotrophically active quantity of a neuronotrophic factor prepared according to the process of claim 1 or 2 and a pharmaceutically acceptable excipient or diluent vehicle.
Claim 27
Dependent Claim
22 words
A pharmaceutical composition according to any of claims 21 to 23, further including at least one ganglioside or at least one phospholipid.
Claim 28
Dependent Claim
28 words
A method for the treatment of neuropathological conditions which comprises administering to a patient an effective amount of a neuronotrophic factor according to claim 11 or claim 14.
Claim 29
Dependent Claim
43 words
The method according to claim 25, wherein the neuropathological condition is a traumatic lesion, a chemical lesion, a vasal lesion, a vasal deficit, a lesion induced by infection or a lesion induced by inflammation or induced by a tumor in the nervous system.
Claim 30
Dependent Claim
16 words
The method according to claim 25, wherein the neuropathological condition is aging of the nervous system.
Claim 31
Dependent Claim
24 words
The method according to claim 25, wherein the neuropathological condition is a chronic neurodegenerative disease or a chronic immunological disease affecting the nervous system.
Claim 32
Dependent Claim
22 words
The method according to claim 25, wherin said neuronotrophic factor is prepared according to the process of claim 1 or claim 2.
Claim 33
Dependent Claim
13 words
The method according to claim 27, wherein the neuropathological condition is Alzheimer's disease.
Claim 34
Dependent Claim
27 words
The method according to claim 26 or claim 27, wherein the neuropathological condition is a chronic neurodegenerative disease, or a chronic immunological disease, affecting the nervous system.
Claim 35
Dependent Claim
19 words
The method according to claim 28, wherein the neuropathological condition is Parkinson's disease, Huntington's chorea or amylotrophic lateral sclerosis.
Claim 36
Dependent Claim
13 words
The method according to claim 28, wherein the neuropathological condition is multiple sclerosis.
Claim 37
Dependent Claim
22 words
The method according to claim 27, wherein said neuronotrophic factor is prepared according to the process of claim 1 or claim 2.
Claim 38
Dependent Claim
24 words
The method according to claim 25, wherein said neuronotrophic factor is administered in association with at least one ganglioside or at least one phospholipid.
References Cited (Backward Citations)
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