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Title:
USE OF A WITHANIA EXTRACT FOR THE TREATMENT OF DEMYELINATING DESEASES
Document Type and Number:
WIPO Patent Application WO/2017/060750
Kind Code:
A1
Abstract:
The invention relates to the use of a composition from a plant extract of Withania somnifera, to treat or limit development of pathologies of the myelin, including multiple sclerosis disease.

Inventors:
RABHI, Chérif (4 impasse Jean Perrin, Bretigny Sur Orge, F-91220, FR)
CARIEL, Léon (19 rue Linné, Paris, F-75005, FR)
OUAZZANI, Jamal (18 avenue Nationale, Massy, F-91300, FR)
ARCILE, Guillaume (13 résidence Le Blois du Roi, Les Ulis, F-91940, FR)
Application Number:
IB2015/001979
Publication Date:
April 13, 2017
Filing Date:
October 05, 2015
Export Citation:
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Assignee:
ETHNODYNE (151 Boulevard Haussmann, Paris, Paris, F-75008, FR)
CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUE (CNRS) (3 rue Michel Ange, Paris, F-75016, FR)
International Classes:
A61K36/81; A61K36/062; A61K36/47; A61K36/80; A61P21/00; A61P25/28
Domestic Patent References:
WO2014202469A12014-12-24
WO2014202469A12014-12-24
Foreign References:
US20110230551A12011-09-22
Other References:
GEETA MAKER-CLARK ET AL: "Integrative therapies for multiple sclerosis", DISEASE-A-MONTH., vol. 59, no. 8, 1 August 2013 (2013-08-01), US, pages 290 - 301, XP055250173, ISSN: 0011-5029, DOI: 10.1016/j.disamonth.2013.03.017
CHIHIRO TOHDA: "ChemInform Abstract: Overcoming Several Neurodegenerative Diseases by Traditional Medicines - The Development of Therapeutic Medicines and Unraveling Pathophysiological Mechanisms", CHEMINFORM, vol. 39, no. 52, 23 December 2008 (2008-12-23), DE, pages 1159 - 1167, XP055250168, ISSN: 0931-7597, DOI: 10.1002/chin.200852253
NAKAYAMA ET AL: "Withanoside IV improves hindlimb function by facilitating axonal growth and increase in peripheral nervous system myelin level after spinal cord injury", NEUROSCIENCE RESEARCH, ELSEVIER, SHANNON, IR, vol. 58, no. 2, 1 June 2007 (2007-06-01), pages 176 - 182, XP022100926, ISSN: 0168-0102, DOI: 10.1016/J.NEURES.2007.02.014
CHÉRIF RABHI ET AL: "Antiangiogenic-Like Properties of Fermented Extracts of Ayurvedic Medicinal Plants", JOURNAL OF MEDICINAL FOOD, vol. 18, no. 9, 1 September 2015 (2015-09-01), US, pages 1065 - 1072, XP055250185, ISSN: 1096-620X, DOI: 10.1089/jmf.2014.0128
CHARLES ET AL., PNAS, vol. 97, 2000, pages 7585 - 7590
BRADL. M.; H. LASSMANN., ACTA NEUROPATHOL, vol. 19, 2010, pages 37 - 53
Attorney, Agent or Firm:
CASALONGA (Casalonga & Associes, 8 avenue Percier, Paris, F-75008, FR)
Download PDF:
Claims:
CLAIMS

1 . Composition containing a Withania somnifera extract for its use to treat or limit devel opm ent of demyelinating di seases in a mammal, preferably a human.

2. Composition according to claim 1 , in which the Withania somnifera extract has been fermented by its incubation with a filamentous fungus in a suitable environment.

3 . Composition according to claim 1 or 2, in which the fermentation is carried out with a filamentous fungus of the family Cordycipitaceae, preferably from the genus B eauveria and, more particularly, Beauveria bassiana.

4. Composition according to any of claims 1 to 3 , further containing at least one extract from the following plants : Emblica officinalis, Bacopa monnieri, Punica granatum, Curcuma longa, Piper longum, or Calendula officinalis .

5. Composition according to any of claims 1 to 3 , further containing an extract of Emblica officinalis and an extract of Bacopa monnieri.

6. Composition according to any of claims 1 to 5 , compri sing a quantity by weight of Withania somnifera of between 5 and 100 g/L of Withania somnifera, and preferably 20 g/L .

7. Composition according to any of claims 1 to 6, compri sing a quantity by weight of Withania somnifera at a concentration of 20 g/L, of Emblica officinalis at a concentration of 1 5 g/L and of Bacopa monnieri at a concentration of 1 5 g/L.

8. Composition according to any of claims 1 to 7, for its use to treat or limit development of multiple sclerosi s, acute di ssemi n ated encephalomyelitis, adrenol eukody strophy, adrenomyeloneuropathy, Leber' s Hereditary Optic Atrophy and related mitochondrial di sorders, HTLV-associated Myelopathy and di seases linked to demyelination of PNS nerves .

9. Composition according to any of claims 1 to 8, for its use to treat or limit development of multiple sclerosi s, wherein the treatment results in one or more clinical outcomes, compared to MS subjects no treated with the composition, selected from the group consisting of:

(a) decrease in MS disease progression,

(b) decrease in M.S disease severity;

(c) decrease in nerve cell demyelination:

(d) decrease in frequency or severity of relapsing MS attacks;

(e) decrease in MS clinical symptoms;

(f) healing of damaged nerve tissue;

(g) increase in remyelination of demyelinated nerves in the central nervous system

(h) protection of damaged nerve tissue from further disease activity

(i) promoting neuronal outgrowth in the central nervous system; and

(j) decrease in disability caused by MS.

10. A method of treating or limiting development of a demyelinating disease in a subject, comprising administering to a subject a therapeutic amount of a plant extract composition, such that said demyelinating diseasee in a subject is treated or limited, wherein said composition contains a plant extract of Withania somnifera according to claims 1 to 8.

11. The method of claim 10, wherein said disease is of multiple sclerosis, acute disseminated encephalomyelitis, adrenoleukodystrophy, adrenomyeloneuropathy, Leber's Hereditary Optic Atrophy and related mitochondrial disorders, HTLV-associated myelopathy and diseases linked to demyelination of PNS nerves.

12. The method according to any of claims 10 to 11, wherein said subject is a human.

13. The method of any of claims 10 to 12, wherein the treatment results in one or more clinical outcomes, compared to MS subjects no treated with the composition, selected from the group consisting of:

(a) decrease in MS disease progression; (b) decrease in MS disease severity;

(c) decrease in nerve cell demyelination:

(d) decrease in frequency or severity of relapsing MS attacks;

(e) decrease in MS clinical symptoms,

(f) healing of damaged nerve tissue,

(g) increase in remyelination of demyelinated nerves in the central nervous system

(h) protection of damaged nerve tissue from further disease activity

(i) promoting neuronal outgrowth in the central nervous svstem; and

(j) decrease in disability caused by MS.

14. The method according to any one of claims 10 to 13, wherein the therapeutic compound is administered orally or parenterally.

15. The method according to any one of claims 10 to 14, wherein said therapeutic compound is administered in a pharmaceutically ptable vehicle.

AMENDED CLAIMS

received by the International Bureau on 25 April 2016 (25.04.2016)

1. Composition containing a Withania somnifera extract fermented by its incubation with a filamentous fungus in a suitable environment for its use to treat or limit development of demyelinating diseases in a mammal, preferably a human.

2. Composition according to claim 1 , in which the fermentation is carried out with a filamentous fungus of the family Cordycipitaceaei preferably from the genus Beauveria and, more particularly, Beauveria bassiana.

3. Composition according to any of claims 1 to 2, further containing at least one extract from the following plants: Emblica officinalis, Bacopa monnieri, Punica granatum, Curcuma longa, Piper longum, or Calendula officinalis .

4. Composition according to any of claims 1 to 2, further containing an extract of Emblica officinalis and an extract of Bacopa monnieri.

5. Composition according to any of claims 1 to 4, comprising a quantity by weight of Withania somnifera of between 5 and 100 g/L of Withania somnifera, and preferably 20 g/L.

6. Composition according to any of claims 1 to 5, comprising a quantity by weight of Withania somnifera at a concentration of 20 g/L, of Emblica officinalis at a concentration of 15 g/L and of Bacopa monnieri at a concentration of 15 g/L.

7. Composition according to any of claims 1 to 6, for its use to treat or limit development of multiple sclerosis, acute disseminated encephalomyelitis, adrenoleukodystrophy, adrenomyeloneuropathy, Leber's Hereditary Optic Atrophy and related mitochondrial disorders, HTLV-associated Myelopathy and diseases linked to demyelination of PNS nerves.

8. Composition according to any of claims 1 to 7, for its use to treat or limit development of multiple sclerosis, wherein the treatment results in one or more clinical outcomes, compared to MS subjects no treated with the composition, selected from the group consisting of:

(a) decrease in MS disease progression;

(b) decrease in MS disease severity;

(c) decrease in nerve cell demyelination:

(d) decrease in frequency or severity of relapsing MS attacks;

(e) decrease in MS clinical symptoms;

(f) healing of damaged nerve tissue;

(g) increase in remyelination of demyelinated nerves in the central nervous system

(h) protection of damaged nerve tissue from further disease activity

(i) promoting neuronal outgrowth in the central nervous system; and

(j) decrease in disability caused by MS.

9. A method of treating or limiting development of a demyelinating disease in a subject, comprising administering to a subject a therapeutic amount of a plant extract composition, such that said demyelinating diseasee in a subj ect is treated or limited, wherein said composition contains a plant extract of Withania somnifera according to claims 1 to 7.

10. The method of claim 9, wherein said disease is of multiple sclerosis, acute disseminated encephalomyelitis, adrenol eukodystrophy, adrenomyeloneuropathy, Leber's Hereditary Optic Atrophy and related mitochondrial disorders, HTLV-associated myelopathy and diseases linked to demyelination of PNS nerves.

1 1 . The method according to any of claims 9 to 1 0, wherein said subject is a human.

12. The method of any of claims 9 to 1 1 , wherein the treatment results in one or more clinical outcomes, compared to MS subjects no treated with the composition, selected from the group consisting of:

(a) decrease in MS disease progression;

(b) decrease in MS disease severity; (c) decrease in nerve cell demyelination:

(d) decrease in frequency or severity of relapsing MS attacks;

(e) decrease in MS clinical symptoms;

(f) healing of damaged nerve tissue;

(g) increase in remyelination of demyelinated nerves in the central nervous system

(h) protection of damaged nerve tissue from further disease activity

(i) promoting neuronal outgrowth in the central nervous system; and

(j) decrease in disability caused by MS.

13. The method according to any one of claims 9 to 12, wherein the therapeutic compound is administered orally or parenterally.

14. The method according to any one of claims 9 to 13 , wherein said therapeutic compound is administered in a pharmaceutically acceptable vehicle.

Description:
Use of a withania extract for the treatment of demyelinating diseases The invention relates to the use of a composition from a plant extract of Withania somnifera, to treat or limit development of pathologies of the myelin, including multiple sclerosi s di sease.

Multiple sclerosi s (MS), al so known as di sseminated scl erosi s or encephalomyelitis di sseminata, i s an inflammatory di sease in whi ch the insulating covers of nerve cells in the brain and spinal cord are damaged. Thi s damage di srupts the ability of parts of the nervous sy stem to communicate, resulting in a wide range of signs and symptoms, including physical, mental, and sometimes psychiatri c problems. The name multiple sclerosis refers to scars (sclerae— better known as plaques or lesions) in particular in the white matter of the brain and spinal cord.

MS takes several form s, with new symptoms either occurring in isolated attacks (relapsing forms) or building up over time (progressive forms) . Between attacks, symptoms may di sappear completely. However, permanent neurological problems often occur, especially as the di sease advances.

Multiple sclerosi s i s the mo st common autoimmune di sorder affecting the central nervous system. As of 2008, between 2 and 2.5 million people are affected globally with rates varying widely in different regions of the world and among different populations . In 201 3 20, 000 people died from MS . The di sease usually begins between the ages of 20 and 50 and i s twice as common in women as in men.

The cause of MS i s not clear, but its pathology consi sts of immune infiltration into the central nervous system (CNS), inflammation, demyelination and finally axonal degeneration. Proposed causes for thi s include genetics and environmental factors such as viral infections. MS i s usually diagnosed based on the presenting signs and symptoms and the results of supporting medi cal tests . Myelination in the CNS involves sequential developmental processes in which precursors of oligodendrocytes (OPCs) migrate, proliferate, and differentiate into newly formed oligodendrocytes (OL), after which those oligodendrocytes selected by target-dependent survival mechanisms wrap myelin membrane around the axons to form the sheath. Each oligodendrocyte can myelinate many axons, with the number of wraps proportional to the axon diameter and regulated tightly by reciprocal signaling between oligodendrocyte and axons.

The development of therapies to promote remyelination in MS is a key research aim, to both aid restoration of electrical impulse conduction in nerves and provide neuroprotection, reducing disability in patients.

So far, there is no known cure for multiple sclerosis. Treatments attempt to improve function after an attack and prevent new attacks.

There are currently a number of disease-modifying medications approved for use in relapsing MS, which includes relapsing-remitting MS and secondary progressive MS. These include interferon beta 1-a, interferon beta 1-b, glatiramer acetate, mitoxantrone and Fingolimod.

Immunosuppressants or cytotoxic agents are used in some subjects after failure of conventional therapies. Other therapeutic approaches include symptomatic treatment which refers to all therapies applied to improve the symptoms caused by the disease and treatment of acute relapses with corticosteroids. While steroids do not affect the course of MS over time, they can reduce the duration and severity of attacks in some subjects.

Other demylinating diseases comprise acute disseminated encephalomyelitis. This disease is characterized by perivascular CNS demyelination, which can occur spontaneously but usually follows a viral infection or very rarely, bacterial vaccination, suggesting an immunologic cause.

The demylinating diseases comprise also the adrenoleukodystrophy/adrenomyeloneuropathy, which are X-linked recessive metabolic disorders characterized by adrenal gland dysfunction and widespread demyelination of the nervous system. Adrenoieukodystrophy occurs in young boys, and adrenomyeloneuropathy in adolescents. Mental deterioration, spasticity, and blindness may occur.

Other demyiinating diseases, like Leber's Hereditary Optic

Atrophy and related mitochondrial disorders are characterized primarily by bilateral loss of central vision, usually affecting males in their late teens or early twenties. Leber's hereditary optic atrophy- resembles the optic neuritis in MS.

HTLV-associated Myelopathy is a slowly progressive spinal cord disease associated with infection by the human T-cell lymphotrophic virus, and is characterized by spastic weakness of both legs.

For the CNS demyelinating diseases:

* Tabes Dorsaiis

• Optic Neuritis

8 Vitamin B12 deficiency

¾ Central Pontine Myelinoiy sis

s Devic's disease

· Transverse Myelitis

• Leukodystrophies

" Progressive Multifocal Leukoencephalopathy

For Peripheral Nervous System

Following are the diseases that occur due to demyelination of peripheral nervous system (PNS) nerves:

• Guillain-Barre syndrome, along with Chronic Inflammatory Demyelinating Polyneuropathy

8 Charcot-Marie-Tooth Disease

• Anti-Myelin Associated Glycoprotein (MAG) peripheral neuropathy

• Copper deficiency- It has been reported that extracts of Withania somnifera,

Emblica officinalis and Bacopa monnieri show anti-angiogenic activity. However, the extracts of these plants were not used because of the high toxicity related to the extracts obtained, and, in particular, the extract from Withania somnifera.

Surpri singly, the applicant has found that, by combining an extraction step and a fermentation step using filamentous fungi on the extracts of the plant Withania somnifera, it is possible to use the detoxified extract to treat MS and other pathologies of the myelin.

The purpose of the invention i s therefore to use a non-toxic composition based on extracts of Withania somnifera, having a protective effect against demyelinating di seases to treat or limit development of MS and related di sorders of the myelin.

Other obj ects, features, aspects and advantages of the invention will appear more clearly on reading the description and examples that follow:

Figure 1 : Effect of Plant extract WEB-2 (5 μg/mL) on OPC number after 12 days of treatment. Data were expressed as percentage of control as mean ± SEM ( 1 00% = no plant extract).

Figure 2 : Effect of Plant extract WEB-2 (5 μg/mL) on OL number and MAG expression after 1 8 days of treatment. Data were expressed as percentage of control as mean ± SEM ( 100% = no plant extract) .

The invention is directed to the use of a composition containing a Withania somnifera extract for its use to treat or limit development of demylinating di seases in a mammal . Preferably, the mammal i s a human.

Preferably, the Withania somnifera extract has been fermented by its incub ation with a filamentous fungus in a suitable environment.

The Withania somnifera plant i s obtained from India. The root of thi s plant i s marketed by Alp Erbo (Marseille) .

The process of production of extracts according to the invention can be found in WO 20 14/202469. Briefly, the plants are fermented in presence of a filamentous fungus of the family Cordycipitaceae, preferably the genus B eauveria. More preferably, the filamentous fungus is derived from the strain Beauveria bassiana, more particularly the strain having reference ATCC 71 59. Preferably, the roots of the plant are used.

The controlled fermentation detoxifies the Withania Somnifera extract by a series of biocatalysi s of various molecules contained in this extract and, more particularly, the chemical family of withanolide aglycones, the sub stances mainly responsible for the toxi city of the extract.

The term " detoxification" i s used to mean elimination by the microorgani sm of potentially toxic molecul es in the medium.

Preferably, after the fermentation, filtration, the medium i s then subj ected to sterili sation steps, preferably by ultrafiltration, in order to obtain the solution which constitutes the plant extract.

The plant extract of the invention contains Withania somnifera but may al so contain at least one of the following extracts : Emblica officinalis, originating from India and marketed by Infrag, B engalore), Bacopa monnieri (India) marketed by Alp Erb o (Marseille), Punica granatum (China) (Shanghai Brightol International Co, Ltd (Shanghai), Curcuma longa (India) (Omnipharm, Chambery), Piper longum (Thailand) (Omnipharm, Chamb ery), or Calendula officinalis (China) (Shanghai Brightol International Co, Ltd (Shanghai), using the same procedure), by independent extraction steps for each plant extract used in the reali sation of the said preparation.

Advantageously, the composition used in thi s invention includes, by weight, between 5 and 1 00 g/L oiWithania somnifera, preferably 20 g/L . Preferentially, thi s composition al so includes one of the following extracts, expressed by weight:

- between 5 and 100 g/L of Emblica officinalis, preferably 1 5 g/L,

- between 5 and 100 g/L of Bacopa monnieri, preferably 1 5 g/L,

- between 5 and 50 g/L of Punica granatum, preferably 1 0 g/L,

- between 5 and 250 g/L of Curcuma longa, preferably 20 g/L,

- between 20 and 50 mg/L of Piper longum, preferably 30 mg/L, - between 5 and 50 g/L of Calendula officinalis, preferably 1 0 g/L,

Preferably, the composition used in thi s invention compri ses an extract of the plants Withania somnifera, Emblica officinalis and Bacopa monnieri. More preferably, the composition according to the invention comprises a quantity by weight of Withania somnifera at a concentration of 20 g/L, of Emblica officinalis at a concentration of 1 5 g/L and of Bacopa monnieri at a concentration of 1 5 g/L .

The compositions according to the invention are used to treat or limit development of multiple sclerosi s, acute di sseminated encephalomyel iti s, adrenoleukodystrophy, adrenomyel oneuropathy, Leber' s Hereditary Opti c Atrophy and rel ated mitochondri al di sorders, HTLV-associated Myelopathy, and di seases linked to demyelination of peripheral nervous sy stem (PNS) nerves.

In certain embodiments of the invention, the methods and compositions reduce the progression of MS in particular.

As used herein, "treating" MS means providing any cli ni cal benefit to a subj ect with MS . The clini cal benefit may be temporary or long-lasting. In various non-limiting embodiments, the treatment results in one or more clinical outcome sel ected from the group consisting of:

(a) decrea se in MS di sease progression;

(b) decrease in MS di sease severity;

(c) decrease in n erve cell demyelination:

(d) decrease in frequency or severity of relapsing MS attacks;

(e) decrease in MS clinical symptoms;

(f) heal ing of damaged nerve tissue (neuro-restoration);

(g) increase in remyelination of demyel inated nerves i n the central nervous system (neuro-restoration) ;

(h) protection of damaged nerve tissue from further di sease activity (neuroprotection) :

(i) promoting neuronal outgrowth (neuro-regeneration) in the central nervous sy stem; and

(j) decrease in di sability caused by MS . As used herein, "limiting development" of MS means providing a limitation in development of symptoms or disease in a subject that is at risk of developing MS. Exemplary subjects at risk of MS include, but are not limited to subjects with a relative (identical twin, non- identical twin, sibling, parent, etc.) that has MS and subjects that, have suffered a clinically isolated syndrome (CIS), which is a subject's first neurological episode, caused by inflammation or demyelinisation of nerve tissue.

In some embodiments, there is a method of treating a demyelinating disease in an individual, comprising the step of delivering to the individual a therapeutic amount of a plant extract composition, such that said demyelinating disease in a subject is treated or its development limited, wherein said composition contains a non toxic plant extract of withania somnifera.

The demyelinating diseases diseases comprise multiple sclerosis, acute disseminated encephalomyelitis, adrenoleukodystrophy, adrenomyeloneuropathy, Leber's Hereditary Optic Atrophy and related mitochondrial disorders, HTLV-associated Myelopathy and diseases linked to demyelination of PNS nerves.

Preferably, the subject is a human.

The composition according to the invention is formulated for oral or parenteral administration.

A person skilled in the art of pharmaceutical formulation will implement the various useful forms for administration of the compositions and/or supplements of the invention. The compositions may be in liquid, gel, emulsion, solid or injectable form.

The composition used may additionally include suspensions, emulsions, syrups containing conventionally used inert diluents, and possibly other substances such as wetting agents, sweeteners, preservatives, thickeners, colourings or any other substance known to a person skilled in the art suitable for oral administration, in particular ((sodium sorbate (E201) (Sigma- Aldrich), anthocyanin (E163) (FBC Industries, USA), sodium metabisulphite (E223) (Sigma- Aldrich), alpha-tocopherol (E307) (FBC Industries, USA). The composition used may al so compri se solvents or other excipients such as water, propylene glycol, vegetable oil s or other suitabl e organic solvents.

The term " excipient" i s used to mean any compound which does not interfere with the effectiveness of the biological activity of the composition according to the invention, and which i s not toxic to the host to which it is admini stered .

The composition used may al so contain adj uvants, such as wetting agents, isotoning agents, emul sifiers, salts or any other sub stances known to a person skilled in the art that can be used as adj uvants (Polydimethyl siloxane, polyvinyl alcohol (PVA), hydrogel s (Carbopol), polyvinylpyrrolidone, hydroxypropyl cellulo se (HPC), poloxamer 1 88, EDTA, chlorobutanol) (Lubrizol, France, Dow Corning, USA) .

Advantageously, the composition may compri se other sub stances such as vitamins, mineral salts, a pharmaceutically acceptable vector, stabili sers, antioxi dants, or any other sub stance known to a person skilled in the art and intended to be integrated into a drug.

Preferably, the composition i s liquid, orally admini strable and contains at least a non-toxic extract of Whitania somnifera, some preservatives, vitamins, water and salt.

More preferably, the preservatives are potassium sorbate or benzoate. The vitamin is riboflavin (vitamin B2) .

The therapeutic composition used in the method of the invention is admini stered in a pharmaceutical ly acceptable vehicl e.

The terms "pharmaceutically acceptable vehicle" is used to mean any vehicle which does not interfere with the effectiveness of the biological activity of the composition according to the invention and which i s not toxic to the host to which it is admini stered.

The composition obtained i s usable as a medicinal product for a mammal, and more particularly for humans, to assi st in the treatment or limitation of development of demyelinating di seases and in particular MS . The term " medicinal product" i s used to mean a product containing an accurate dose of said preparation according to European directive 65/65/EC, namely any sub stance or composition describ ed as possessing curative or preventive properties with respect of human or animal di sease. For example, the medicinal product containing sai d preparation at therapeutic doses can be admini stered orally as a cap sule or a tablet, or inj ected via any other route to confer the beneficial effects .

An appropriate dosage of the therapeutic composition can be determined by one of skill in the art, taking into consideration the findings described herein together with typical factors such as the body mass of the patient, the physical condition of the patient, and so on. The dosage should contain the therapeutic composition in an amount that is effective for treating or limiting development of demyelinating di seases and in particular MS .

The drug can be admini stered daily, weekly, or on an intermittent basis. For exampl e, the drug can be admini stered for three weeks on, followed by one week off, or for two weeks on, followed by one week off, or under other dosing schedul es as can be determined by one skilled in the field.

The particular dose selected will depend upon the mode of admini stration and dosing regimen selected . One preferred schedule i s a once daily oral dosing schedule. When longer periods of time are prescribed between each application (typically the case for i. v. admini stration), each unit dose may be larger than when daily dosages are provided.

The daily dose of the compositions used may vary according to the needs and severity of symptoms of the patient and according to the route . Typically, the daily dose i s between 10 mg/raL and 300 mg/mL of the composition after fermentation.

Preferably, the daily dose for an adult human i s between 30 and 100 mg/mL of the composition after fermentation.

The present invention will be expl ained in further detail by way of non-limiting examples below, which make reference to the appended drawings . The following methods were used in the experiments described in the examples that follow the description of the methods .

Example 1 : Composition WEB- 1 b efore fermentation

The composition WEB- 1 contains a commercial extract of

Withania Somnifera at a concentration of 20 g/L, of Emblica officinalis at a concentration of 1 5 g/L, of Bacopa monnieri at a concentration of 1 5 g/L.

A solution of 100 mL i s made in water. After lyophilization,

3 .8 g of a beige powder i s obtained.

Example 2 : Strain of filamentous fungus Beauveria bassiana The strain Beauvaria Bassiana (reference ATCC 71 59) has been cultivated in a medium containing 0, 5 g/L KH 2 P0 4 ; 1 g/L

KH 2 P0 4 ; 1 g/L MgS 0 4 ; 2 g/L NaN0 3 ; 0, 5 g/L KC1 ; 0, 02 g/L FeS0 4 ;

30 g/L glucose (all reagents from Sigma- Aldrich, France) and 1 0 g/L of corn steep liquor (Roquette, France) .

The culture was then agitated at 200 rotations per minute, for

72 hours at 27 °C . It was then filtered by non-sterile methods on a filter paper to separate the fungal biomass from the culture medium .

The fungal biomass was then washed thoroughly with water.

Example 3 : Composition WEB-2 used in the invention

The composition WEB- 1 as in example 1 i s added to the fresh fungal biomass of example 2 using 60 g of biomass per liter of composition WEB- 1 containing 50g of glucose.

After incubation, thi s seeded composition was agitated at 200 rpm for 5 days at a temperature of 27 ° C .

After 5 days, the incubation medium was filtered on a filter paper, the samples for HPLC assay were al so filtered using a 0.45 micron filter (Ait-France, ref: SFNY 013045N) .

The browni sh solution obtained which was then lyophilized during 5 days to produce dried beige powder. Example 4 : Composition WE-2 used in the invention

The composition WE- 1 contains commercial extracts of Withania Somnifera at a concentration of 20 g/L, and of Emblica officinalis at a concentration of 1 5 g/L.

To 100 mL of such a solution, are added 5 g of glucose and 6 g of biomass of example 2.

After having treated and lyophilized the solution like in example 3 , 4. 1 3 g of a beige powder i s obtained.

The markers identified in the composition WE-2 were

Withanoside IV, Withanoside VI and gallic acid.

Example 5 : Composition WB-2 used in the invention

The composition WB- 1 contains an extract of Withania Somnifera at a concentration of 20 g/L, and of Bacopa Monnieri at a concentration of 1 5 g/L.

To 100 mL of such a solution, are added 5 g of glucose and 6 g of biomass of example 2.

After having treated and lyophilized the solution like in example 3 , 2, 62 g of a beige powder i s obtained.

The markers identified in the composition WB-2 were Withanoside IV, Withanoside VI, B acoside A3 , B acopaside X and Bacopa, saponin C . Example 6 : Composition BE-2 used in the invention

The composition BE-2 contains an extract of Bacopa Monnieri at a concentration of 1 5 g/L, and of Emblica officinalis at a concentration of 1 5 g/L.

To 100 mL of such a solution, are added 5 g of glucose and 6 g of biomass of example 2.

After having treated and lyophilized the solution like in example 3 , 2, 62 g of a beige powder i s obtained.

The markers identified in the composition BE-2 were Bacopaside X, B acopa, saponin C and gallic acid. Example 7 : Composition WEB-4 according to the invention The composition WBE-4 contains an extract of Withania

Somnifera at a concentration of 40 g/L, of Bacopa Monnieri at a concentration of 1 5 g/L, and of Emblica officinalis at a concentration of 1 5 g/L .

To 100 mL of such a solution, are added 5 g of glucose and 6 g of biomass of example 2.

After having treated and lyophilized the solution like in example 3 , 4.23 g of a beige powder i s obtained.

Example 8 : Composition WEB-6 used in the invention

The composition WEB-6 contains an extract of Withania

Somnifera at a concentration of 20 g/L, of Bacopa Monnieri at a concentration of 1 5 g/L, and of Emblica officinalis at a concentration of 30 g/L .

To 100 mL of such a solution, are added 5 g of glucose and 6 g of biomass of example 2.

After having treated and lyophilized the solution like in example 3 , 4.22 g of a beige powder i s obtained.

Example 9 : Composition WEB-8 used in the invention

The composition WEB-8 contains an extract of Withania

Somnifera at a concentration of 20 g/L, of Bacopa Monnieri at a concentration of 30 g/L, and of Emblica officinalis at a concentration of 1 5 g/L .

To 100 mL of such a solution, are added 5 g of glucose and 6 g of biomass of example 2.

After having treated and lyophilized the solution like in example 3 , 3 .76 g of a beige powder i s obtained.

Example 10 : in vitro MS model A reproducible in vitro myelination model based on primary cocultures of central neurons and oligodendrocytes culturing in 96- well plate is used and adapted to high throughput screening. a) Culture of neurons/oligodendrocytes

Neurons/oligodendrocytes were cultured as previously described by Charles et al., 2000. PNAS 977585-7590.

Briefly, pregnant Wistar female rats of 17 days gestation were killed by cervical dislocation (Janvier Labs, France) and the foetuses removed from the uterus. The forebrains were removed and placed in ice-cold medium of Leibovitz (L15; Pan Biotech, Germany) containing 2% of Penicillin-Streptomycin (Batch 1451013, PanBiotech) and 1% of bovine serum albumin (BSA) (Batch K180713, Pan Biotech). Cortexes were dissociated by trypsinisation for 20 min at 37°C, with Trypsin EDTA IX (Batch 7310713, PanBiotech). The reaction was stopped by the addition of Dulbecco's modified Eagle's medium (DMEM) (Batch 9710913, PanBiotech) containing DNAase I grade II at 0.1 rag/mL (Batch H131108, PanBiotech) and 10% of foetal calf serum (Batch 41Q7218K, Invitrogen, France). Cells were then mechanically dissociated by 3 passages through a 10 mL pipette. Cells were centrifuged at 180 x g for 10 min at 4°C temperature on a layer of BSA (3.5%)) in L15 medium. The supernatant was discarded and cells of the pellet were re-suspended in DMEM containing 10%> of FCS. Cells were then centrifuged at 515 x g for 10 min at 4°C. The supernatant was discarded and cells of pellet were re-suspended in a culture medium consisting of Neurobasal (Batch 1625353, Invitrogen) supplemented with 2% of B27 (Batch 1618508, Invitrogen), 2 mM of L-glutamine (Batch 6620314, PanBiotech), 2% of PS solution, 1 % of foetal calf serum (FCS) and 10 ng/mL of platelet-derived growth factor (PDGF- AA) (Batch H131205, PanBiotech). Viable cells were counted in a Neubauer cytometer using the trypan blue exclusion test. The cells were seeded at a density of 20000 cells/well in 96 well-plates pre- coated with poly-L-lysine (Batch 3102256, Beckton-Dickinson, France) and laminin (Batch 083M4034V, Sigma- Aldrich France). The plant extract (500 μ§/ιηΙ., 50 μ§/ιηΙ., or 5 μ§/ιηΙ.), was solved and diluted in culture medium and then pre-incubated with neurons lh after the cell plating. The plates were maintained at 37°C in a humidified incubator, in an atmosphere of air (95%)-C02 (5%). Half of the medium was changed every 2 days with fresh medium in presence or absence of plant extract.

The following conditions were assessed:

Plate 1 for Day 12 (OPC evaluation)

- Control (vehicle)

- Plant extract WEB-2 (500 μg/mL)

- Plant extract WEB-2 (50 μg/mL)

- Plant extract WEB-2 (5 μg/mL)

Plate 2 for Day 18 (OL evaluation)

- Control (vehicle)

- Plant extract WEB-2 (500 μg/mL)

- Plant extract WEB-2 (50 μg/mL)

- Plant extract WEB-2 (5 μg/mL) b) immunostaining of cells

On days 12 and 18 of culture, cells were fixed by a cold mixture of absolute ethanol 95% (Batch SZBD1470V, Sigma) and acetic acid 5% (Batch SZBD1760V, Sigma) for 5 min. The cells were then permeabilized and non-specific sites were blocked with a solution of phosphate buffered saline (Batch 3010914, PanBiotech) containing 0.1% of saponin (Batch BCBJ8417V, Sigma) and 1% FCS for 15 min at room temperature.

On day 12, cells were incubated with Monoclonal Anti-A2B5 conjugated alexa fluor® 488 produced in mouse (Batch 2281669, Millipore, France) at dilution of 1/200 in PBS containing 1% FCS, 0.1 % saponin, and with anti-MAP-2 antibody produced in chicken (Batch GR180541-3, AbCam; United Kingdom) at dilution of 1/1000 in PBS containing 1% FCS, 0.1 % saponin for 2 h at room temperature. This antibody was revealed with Alexa Fluor 568 goat anti-chicken antibody (Batch 1383072, Molecular probe; France) at the dilution of 1/400 in PBS with 1% FCS, 0.1 % saponin, for 1 h at room temperature.

On day 18, cells were incubated with Monoclonal Anti-MAG antibody produced in mouse (Batch 2301638, Millipore) at dilution of 1/400 in PBS containing 1% FCS, 0.1 % saponin, and with anti-MAP-2 antibody produced in chicken at dilution of 1/1000 in PBS containing 1% FCS, 0.1 % saponin for 2 h at room temperature. These antibodies were revealed with Alexa Fluor 488 goat anti-mouse antibody (Batch 1397999, Molecular probe) at the dilution of 1/400 in PBS with 1% FCS, 0.1 % saponin and Alexa Fluor 568 goat anti-chicken antibody at the dilution of 1/400 in PBS with 1% FCS, 0.1 % saponin, for 1 h at room temperature. c) Results

The immunolabeled cultures were automatically examined with

ImageXpress (Molecular Devices) equipped with a LED at x20 magnification. For each condition, 30 pictures (representing -80 % of the total surface of the well) per well were taken. All images were taken with the same conditions. Number of A2B5 positive cells and number of MAG positive cells were automatically analyzed by using Custom module editor (Molecular Devices). Data were expressed in percentage of control conditions (no plant extract = 100 %). All values were expressed as mean +/- SEM (s.e.mean) of the 6 wells. Graphs and statistical analyses are made on the different conditions (ANOVA followed by Dunnett's test when allowed, using GraphPad Prism software).

Treatment with Plant extract WEB-2 (5 μg/mL) did not show any significant increase of the OPC number after 12 days treatment (figure 1). The slight increase of cell number seemed to be the consequence of a protective effect against the spontaneous apoptosis existing in all cultures.

Treatment with Plant extract WEB-2 (5 μg/mL) showed a significant effect on the number of MAG positive cells as well as in the expression of MAG (area parameter). Myelin Associated Glycoprotein (MAG) is a specific protein of differentiated oligodendrocytes that is heavily expressed in myelinating oligodendrocytes (Bradl. M., & H. Lassmann., 2010. Acta Neuropathol (2010) 119:37-53).

After 18 days of application, WEB-2 showed a large increase in the number of myelinating OL (20 %). More important, these OL showed large amount of MAG protein (60 % more than control condition) (Figures 2a and 2b)

The 2 highest doses of the plant extract showed toxicity after 2-3 days (for 500 μg/mL) or 4-5 days (for 50 μg/mL) of culture.