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WO2006004386A1 - Use of ginkgo biloba extract for the preparation of a medicine that is used to treat parkinson's disease - Google Patents

Use of ginkgo biloba extract for the preparation of a medicine that is used to treat parkinson's disease Download PDF

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Publication number
WO2006004386A1
WO2006004386A1 PCT/MX2004/000042 MX2004000042W WO2006004386A1 WO 2006004386 A1 WO2006004386 A1 WO 2006004386A1 MX 2004000042 W MX2004000042 W MX 2004000042W WO 2006004386 A1 WO2006004386 A1 WO 2006004386A1
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extract
pharmaceutical composition
disease
ginkgo biloba
mpp
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WO2006004386A8 (en
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Patricia ROJAS CASTAÑEDA
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/16—Ginkgophyta, e.g. Ginkgoaceae (Ginkgo family)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/14—Drugs for disorders of the nervous system for treating abnormal movements, e.g. chorea, dyskinesia
    • A61P25/16—Anti-Parkinson drugs

Definitions

  • the present invention is directed to the use of an extract of Ginkgo biloba, to prepare a medicine intended to treat Parkinson's disease. This represents a potential and important alternative for the treatment of Parkinson's disease.
  • the applicant was able to notice the beneficial effects of an extract of Ginkgo biloba in a pharmacological model of the disease, using the neurotoxin 1-methyl-4-phenyl-1, 2,3,6-tetrahydropyridine named hereinafter like MPTP.
  • Parkinson's disease has an incidence of 168 / 100,000 and is a well-known disease especially in the population over the age of 60 years. Symptoms of this disease include uncontrolled movements, tremor at rest, gait disturbances, postural abnormalities, and muscle stiffness.
  • the primary neuropathology associated with this disorder is the progressive and persistent loss of dopaminergic neurons originating from the nigra pars compact substance that project to the striatum. This leads to a substantial decrease in the neurotransmitter dopamine in the nigrostriatal pathway (Jenner, P. 1989. Clues to the mechanism underlying dopamine cell death in Parkinson's disease. J. Neurol. Neurosurg. Psychiatry. (Special suppl): 22-28). The subsequent decrease in the synthesis of dopamine correlates with the onset and severity of the mentioned symptoms. It is important to emphasize that the synthesis of dopamine is determined by the disposition of the activity of the tyrosine hydroxylase to carry out the synthesis of catecholamines including dopamine.
  • Free radicals are highly reactive molecules produced in the body as normal products of metabolism and in large quantities by Ia inflammation, alcoholism, smoking, radiation, etc. Those highly reactive molecules can oxidize other molecules in the body, making them unstable and potentially very harmful. Free radicals are highly unstable molecules that can cause damage to deoxyribonucleic acid, carbohydrates, lipids, and proteins in the body (Halliwell, B. and Gutteridge, JMC Free you are in biology and medicine. Oxford-Claredon, 1985). Of these effects, one of the most important is lipid peroxidation, the index most commonly used to measure the biological effect of free radicals. This damage known as oxidative stress, is implicated in the pathogenesis of a large number of chronic diseases, including Parkinson's disease.
  • Organisms have developed various defense mechanisms to protect themselves against free radical damage. Those defense mechanisms include antioxidant enzymes, free radical scavengers, and metal chelating agents (Reiter, RT. 1995. Oxidative processes and antioxidative defense mechanisms in the aging brain. FASEB J. 9: 526-533).
  • the antioxidant enzymes are catalase, glutathione peroxidase, and superoxide dismutase. Normally, there is a balance between the generation of free radicals and antioxidant defense systems in vivo. When this balance is altered in favor of the production of free radicals due to the decrease in antioxidant systems or an increase in the generation of free radicals, oxidative stress occurs.
  • Oxidative stress leads to damage to polyunsaturated fatty acids by lipid peroxidation, a chain reaction that results in numerous degradation products (Niké, E., Noguchi, N. and Gotoh, N. 1993. Dynamics of lipid peroxidation and its inhibition by antioxidants. Biochem. Soc. Trans. 21: 313-317).
  • MPTP is referred to as the best experimental model of Parkinson's disease (Gerlach, M., Riederer, P., Przuntek, H., and Youdim, MBH 1991. MPTP mechanisms of neurotoxicity and their implications for Parkinson's disease. Eur J. Pharmacol. Mol. Pharmacol. 208: 273-286).
  • MPTP When MPTP is administered to primates that are not human and mice, it produces hypokinesia and neuronal damage similar to that observed in Parkinson's disease (Heikkila RE, Cabbat FS, Manzino L, Duvoisin RC 1984. Effects of MPTP on nigrostriatal dopamine in mice. Neuropharmacology 23: 711-713).
  • MPP + The 1-methyl-4-phenylpyridine ion that we will hereinafter refer to as MPP + is the largest neurotoxic metabolite of MPTP and is responsible for its neurotoxic effects.
  • the biotransformation of the MPTP to its toxic metabolite MPP + in the brain is initially carried out by the monoamine oxidase B enzyme that we will hereinafter refer to as MAO-B and this generates free radicals (Zang, LY and Misra, HP 1993. Generation of reactive oxygen species during the monoamine oxidase-catalyzed oxidation of the neurotoxicant, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine. J. Biol. Chem. 268: 16504-16512).
  • the monoamine oxidase that we will hereinafter refer to as
  • MAO exists in 2 isoforms A and B.
  • MAO is the largest intracellular enzyme that metabolizes dopamine in the central nervous system.
  • the high oxidation of dopamine by MAO-B in Parkinson's disease may involve Ia production of hydrogen peroxide, a toxic molecule derived from oxygen capable of inducing the formation of free radicals, which could potentially harm dopaminergic neurons of the nigrostriatal pathway (Cohén, G. 1986. Monoamine oxidase, hydrogen peroxide, and Parkinson's disease . Pages 119- 125, in Yahr, MD, and Bergmann KJ (eds.), Advances in Neurplogy. Parkinson 's disease, vol 45, Raven Press, New York)..
  • Parkinson's disease As mentioned, currently the therapy of choice for patients who have Parkinson's disease is through the administration of L-DOPA, which is converted to dopamine in the brain. This pharmacological treatment is initially effective, but it often becomes less effective over time and in many cases the symptoms of the patients get worse. So far there is no clear and effective cure for Parkinson's disease.
  • the present invention represents a potential and important alternative for the therapy used for the treatment of Parkinson's disease. This is anticipated that treatment with an extract of Ginkgo biloba to patients with Parkinson's disease will inhibit or stop the progress of the disease by reducing the degeneration and dysfunction of dopaminergic neurons of the nigrostriatal pathway. Likewise, act as an antioxidant and / or antilipoperoxidant agent, regulate and / or inhibit the activity of the MAO that is generating free radicals; and increase the activity of tyrosine hydroxylase, enzyme required for the synthesis of dopamine.
  • Ginkgo biloba extract is meant that it is at least an extract of the individual compounds that can be obtained by extraction from the Ginkgo biloba tree and in particular a flavonoid compound or a terpene such as a ginkgolide or a bilobalide, or also a mixture.
  • an extract of the EGb 761 type can for example be chosen.
  • EGb 761 extract is a well-defined mixture of active compounds extracted from the leaves of Ginkgo biloba. This is an extract from the leaves of Ginkgo biloba which is substantially free of alkyphenol compounds, has a high content of flavonoid glycosides, and which contains substantially all of the ginkgolides and bilobalides originally present in the leaves.
  • extract of the EGb 761 type is meant an extract of a composition substantially identical to that of the standardized extract EGb 761 defined in the following article: K. Drieu, La presse med ⁇ cale, 31, Sep. 25,
  • extract of type EGb 761 is by Io understood in particular of extracts of Ginkgo biloba comprising 20 to 30% glycosides of flavones, 2.5 to 4.5% of ginkgolides A, B, C, and J, 2 to 4% of bilobalides, less 10% proanthocyanidins and less than 10 ppm, and preferably less than 5 ppm, of compounds of the alkylphenol type, and in particular extracts of Ginkgo biloba comprising approximately 24% flavone glycosides, 3.1% ginkgolides A, B, C and J , 2.9% of bilobalides, 6.5% of proanthocyanidins and less than 1 ppm of compounds of the alkylphenol type.
  • EGb 761 has been used in the clinic for the treatment of cerebrovascular insufficiency, degenerative dementia, peripheral vascular disorders and sensorineural disorders (DeFeudis, FV1998. Gingko biloba extract (EGb761): from Chemistry to the clinic. Pages 401. Ullstein Medical ,
  • a multipurpose action of EGb 761 is responsible for its effectiveness in treat clinical disorders of multifactorial origin.
  • the additive, antagonistic and synergistic effects of the active constituents of EGb 761 probably occur with respect to various molecular target sites in different tissues and organs.
  • the object of this invention is to provide drugs that contain this extract of Ginkgo biloba with a high content of flavonoid glycosides, ginkgolides and bilobalides originally present in the leaves and where there is no substantial danger of allergic reactions, precisely due to the removal of the compounds alkyphenols.
  • the extract in the present invention should contain: - from 20 to 30 percent by weight, in particular 22 to 26 percent by weight of flavonoid glycosides.
  • the Ginkgo biloba extract of the invention can be processed in Ia usual route of pharmaceutical preparations, for example for solutions, coated tablets, tablet preparations or injections.
  • the object of the present invention is the use of an extract of Ginkgo biloba, to prepare a medicine intended to treat Parkinson's disease.
  • the compounds can be administered for several months for treatment.
  • the pharmaceutical compositions comprise a compound of the invention that can be in the form of solids, for example powders, tablets, capsules, liposomes or suppositories.
  • the excipients can be for example calcium phosphate, magnesium stearate, talc, sugars, lactose, dextrin, gelatin, starch, cellulose, methyl cellulose, sodium carboxymethyl cellulose, polyvinylpyrrolidine and wax.
  • compositions containing a compound of the invention may also be present in the form of a liquid, for example, solutions, emulsions, suspensions or syrup.
  • Suitable excipients for liquids can be, for example, water, organic solvents such as glycerol or glycols, as well as their mixtures, in various proportions.
  • the administration of the medicine according to the invention can be topical, oral, parenteral route, by injection, such as intramuscular, subcutaneous, intravenous administration, etc.
  • the daily administration dose of the active ingredients that can be used according to the invention is between 0.1 mg to 1 g depending on the severity of the Parkinson's disease of the patient to be treated.
  • a dose ranging from approximately 5 mg to 600 mg per day may be chosen.
  • FIG. 1 This graph shows that treatment with an extract of Ginkgo biloba type EGb 761 regulates the activity of MAO B in the striatum in parkinsonism induced with MPP + .
  • the results are expressed in ⁇ moles of 4 HOQV / g / h.
  • Each bar represents the average +/- standard error of 4-6 experiments per group.
  • + Statistically different from the MPP + group represented as "saline + MPP + ", p ⁇ 0.05, Tukey test.
  • Figure 2 It is a graph that shows the restoration that produces the extract of Ginkgo biloba type EGb 761 in the levels of dopamine in the striatum in animals that were previously parkinsonized with MPTP.
  • A represents the "saline + saline” group
  • B represents the “saline + EGb 761 group at a dose of 40 mg / kg”
  • C represents the "saline + EGb 761 group at a dose of 120 mg / kg
  • D represents the” MPTP + saline "group
  • E represents the” MPTP + EGb 761 group at a dose of 40 mg / kg
  • F represents the” MPTP + EGb 761 group at a dose of 120 mg / kg ".
  • Examples 3 to 7 were carried out using male mice of the C-57 black strain of 25-30 g in weight. The animals were kept in standard conditions with a cycle of 12: 12h light / dark, 21-22 ° C, relative humidity 40%, with food and water ad libitum.
  • Ginkgo biloba extract type EGb 761 was generously donated by Dr. Willmar Schwabe GmbH & Co. KG in Düsseldorf, Germany. EGb 761 was dissolved in saline and adjusted to a pH of 7.4.
  • Solution for oral administration where 10O ml of solution contain:
  • group I saline intraperitoneally, hereinafter referred to as ip + saline intracerebroventricular, hereinafter referred to as icv
  • group II EGb 761 by ip route + saline solution by icv route
  • group III saline by ip route + MPP + by icv route
  • group IV EGb 761 by ip route + MPP + by icv route.
  • the tissue was homogenized in 2.5 ml of phosphate buffer with a pH of 7.0. 1 ml aliquots of the homogenate were taken and placed in glass tubes with lids. Subsequently I added 3 ml of a chloroform-methanol mixture at a 2: 1 volume / volume dilution. The tubes were capped and the mixture was gently shaken and placed on ice for 30 minutes. The aqueous phase was discarded and 1 ml of the chloroform phase was transferred in quartz cells, 0.1 ml of methanol were subsequently added.
  • Table 1 shows that the administration of extract type EGb 761 did not produce significant increases in the formation of PFL at the different doses tested, which were 2.5, 5 and 10 mg / kg, at 2 h after the administration of MPP + compared to control animals.
  • the lipid peroxidation at 2 h after the administration of MPP + without pretreatment with EGb 761 type extract produced an increase in lipid peroxidation of 72%.
  • the pretreatment with extract type EGb 761 at 10 mg / kg blocked 100% the lipid peroxidation induced by MPP + , that is, the levels of lipid peroxidation are not statistically different from controls treated with saline.
  • Table 1 shows the dose response study of the EGb type extract
  • the degree of protection increased with increasing dose of the EGb 761 type extract.
  • the degree of protection was 65%, 93% and 100% for 2.5, 5 and 10 mg / kg of EGb 761, respectively, when compared to the "saline + MPP + " group.
  • MAO inhibitors deprenil for MAO-B and clorgiline for MAO-A, at a concentration of 500 nM in the final volume and preincubated for 10 min. These were used to measure these MAO isoforms only in the 6h group.
  • the administration of the extract of Ginkgo biloba type EGb 761 to the mice did not produce significant increases in the activity of the total MAO at different times tested of 2, 6, 12 and 24 h, when compared with the group treated with saline.
  • the total activity of the MAO was significantly elevated at 2 and 6 h after its administration, that is, 29% and 30% respectively compared to the control group, p ⁇ 0.05, without pretreatment.
  • Ginkgo biloba extract type EGb 761 no changes in the total activity of the MAO were observed at 12 and 24 h after the administration of MPP + compared to the group treated with saline solution.
  • pretreatment with Ginkgo biloba extract type EGb 761 in the MPP + group prevented the increase in MAO activity in the MPP + group at 6 h, observed without pretreatment. This means that the levels of the activity of the MAO were not statistically different from those of the controls treated with saline solution. Although the pretreatment with the EGb 761 type extract seemed to prevent the increase in the activity of the MAO produced by the MPP + at 2 h, but it was not statistically significant.
  • This table shows the beneficial effects of the regulation and / or inhibition of the enzymatic activity of the total MAO in the striatum by pretreatment with Ginkgo biloba extract type EGb 761 in parkinsonism experimentally induced with MPP + .
  • the results are expressed in activity of the total MAO in ⁇ moles 4-OHQ / g / h.
  • Each group represents the average +/- standard error of the average of 6-8 experiments per group.
  • mice The brains of the mice were immediately removed and the striatum was dissected. A 500 ⁇ l aliquot of 0.1% w / v sodium perchloric-metabisulfite solution was added to the tissue weight and sonicated with a Labsonic Lab-line instruments model system, Melrose Park.IL. Subsequently the samples were centrifuged at 4,000 g for 10 min and the supernatants were kept at -70 0 C until analysis.
  • the activity of the tyrosine hydroxylase was assayed in the striatum using a Perkin-Elmer model series 200 LC high-performance liquid chromatography system, connected to an ESA Coulochem 5100A detector with the following reference electrode potentials of +0.4 V and oxidation of Ia CeWa 1 ZCeWa 2 + 0.03 / + 0.35 V.
  • ESA Coulochem 5100A detector with the following reference electrode potentials of +0.4 V and oxidation of Ia CeWa 1 ZCeWa 2 + 0.03 / + 0.35 V.
  • the calibration curves were built for dopamine, L-DOPA and the concentration was obtained by interpolation of the respective standard curve.
  • the mobile phase consisted of an aqueous phosphate buffer at a pH of 3.1, which contained sodium octyl sufate at 0.2 mM, EDTA at 0.1 mM, methanol 15% volume / volume, at a pH of 2.6.
  • the EGb 761 type extract only does not influence the activity of the tyrosine hydroxylase at all the times tested when it is only compared with the "saline + saline” group.
  • Dopamine concentrations were analyzed according to the previously described method (Larsson LG, Rényi L, Ross SB, Svensson B, and ⁇ ngeby-Móller K. 1990. Different effects on the responses of functional pre- and postsynaptic 5-HT 1A receptors by repeated treatment of rats with the 5-HT-IA receptor agonist 8-OH-DPAT. Neuropharmacology. 29: 85-91). A 500 ⁇ l aliquot of 0.1% w / v sodium perchloric-metabisulfite solution was added to the tissue weight and sonicated with a Labson line brand Labsonic system, Melrose Park.IL.
  • This table shows the increase in tyrosine hydroxylase activity in the striatum by pretreatment with EGb 761-type extract as a protective response in parkinsonism experimentally induced with MPP + .
  • the results are expressed as tyrosine hydroxylase activity in ⁇ g / g of tissue.
  • Each group represents the average +/- standard error of the average of 6-8 experiments per group. * Statistically different from the control group represented as "saline + saline", p ⁇ 0.05, Tukey's test. + Statistically different from the group treated with MPP + represented as "saline + MPP + ", p ⁇ 0.05, Tukey's test.
  • group I saline solution via i.p + saline solution via i.p route
  • group II saline by the i.p + EGb 761 route by the i.p route
  • group III MPTP via ip + saline via i.p .
  • Group IV MPTP via i.p. + EGb 761 via i.p.
  • mice in groups I and Il received saline ip and groups III and IV received MPTP at a dose of 30 mg / kg, ip daily for 5 days.
  • animals in groups Il and IV were administered with EGb 761 type extract at a dose of 40 or 120 mg / kg for 18 days or saline was administered in groups I and III using them as a control group.
  • the animals were sacrificed by cervical dislocation after the last administration of the type extract.
  • EGb 761 The brains were immediately removed and the striatum was dissected, and the dopamine content in the striatum was analyzed by high-performance liquid chromatography as described in Example 6. TABLE 4.
  • This table shows the protection produced by pretreatment with extract type EGb 761 in the levels of dopamine in the striatum in parkinsonism experimentally induced with MPP + .
  • the results are expressed in dopamine content in ⁇ g / g of tissue.
  • Each group represents the average of +/- standard error of the average of 6-8 experiments per group.

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Abstract

The invention relates to the use of Ginkgo biloba extracts and, in particular, Ginkgo biloba extracts that contain 20 - 30 % flavonoid glycosides, 2.5 - 4.5 % ginkgolides A, B, C and J, 2 - 4 % bilobalides, less than 10 % proanthocyanidins and less than 10 ppm alkyphenol-type compounds, in order to prepare a medicament for the treatment of Parkinson's disease. The invention also relates to the treatment of said disease with the aforementioned Ginkgo biloba extract which acts as an antioxidant and/or antilipoperoxidant agent, regulates MAO activity, increases tyrosine hydroxylase activity and increases and restores the dopamine content in MPTP-induced parkinsonism.

Description

USO DE UN EXTRACTO DEL GINKGO BILOBA PARA PREPARAR UNA MEDICINA PARA TRATAR LA ENFERMEDAD DE PARKINSON USE OF A GINKGO BILOBA EXTRACT TO PREPARE A MEDICINE TO TREAT PARKINSON'S DISEASE

Descripción de Ia invenciónDescription of the invention

Campo de Ia invenciónField of the invention

La presente invención esta dirigida al uso de un extracto del Ginkgo biloba, para preparar un medicamento destinado a tratar Ia enfermedad de Parkinson. Esta representa una alternativa potencial e importante para el tratamiento de Ia enfermedad de Parkinson.The present invention is directed to the use of an extract of Ginkgo biloba, to prepare a medicine intended to treat Parkinson's disease. This represents a potential and important alternative for the treatment of Parkinson's disease.

AntecedentesBackground

Es conocido que los extractos de Ginkgo biloba tienen una actividad en el campo cardiovascular y en particular Ia reducción de Ia adhesión plaquetaria; en el sistema nervioso central, en particular una actividad neuroprotectora ó en el sistema neurosensorial, en particular en Ia protección de Ia retina (DeFeudis et al., Ginkgo biloba Extract (EGb 761), Pharmaceutical Activities and Clinical Applications (Elsevier, Paris, 1991). Sus preparaciones han sido objeto de un cierto número de patentes, de las cuales se pueden mencionar las patentes Europeas EP 431 535 y EP 431 536, y Ia patente Americana U.S. Pat. No. 5, 389, 370.It is known that the extracts of Ginkgo biloba have an activity in the cardiovascular field and in particular the reduction of platelet adhesion; in the central nervous system, in particular a neuroprotective activity or in the sensorineural system, in particular in the protection of the retina (DeFeudis et al., Ginkgo biloba Extract (EGb 761), Pharmaceutical Activities and Clinical Applications (Elsevier, Paris, 1991 Its preparations have been the subject of a number of patents, of which the European patents EP 431 535 and EP 431 536, and the American patent US Pat. No. 5, 389, 370 can be mentioned.

Ciertos productos pueden ser usados en el tratamiento de Ia enfermedad de Parkinson. Sin embargo, en Ia actualidad Ia terapia de elección para pacientes que tienen esa enfermedad es a través de Ia administración de β-3, 4- dihydroxifenil- α-alanina conocida como L-DOPA y que así denominaré de aquí en adelante, esta pasa Ia barrera hematoencefálica y es convertida a dopamina en el cerebro (Quinn, N. P. 1990. Levodopa-based therapy. Pages 169-184, in Therapy of Parkinson's disease. Marcel Dekker, New York). Este tratamiento farmacológico es inicialmente efectivo, pero el tratamiento con L-DOPA tiene graves efectos secundarios, como Ia producción de movimientos incontrolados y distonia (Kanazawa, I. 1986. Clinical pathophysilogy of basal ganglia diseases. VoI. 49, pages 65-92, in Vinken, P.T., Bruyn, G.W., Klawans, H. L. (eds): Handbook of Clinical Neurology. Elsevier Science Publishers, New York). Hasta el momento no hay clara y efectiva cura para Ia enfermedad de Parkinson. Ahora el aplicante ha encontrado que un extracto del Ginkgo biloba, también tiene nuevas propiedades farmacológicas, retardando y atenuando el daño dopaminérgico producido en Ia enfermedad de Parkinson.Certain products can be used in the treatment of Parkinson's disease. However, currently the therapy of choice for patients who have this disease is through the administration of β-3, 4- dihydroxyphenyl-α-alanine known as L-DOPA and which I will call hereinafter, this happens The blood-brain barrier and is converted to dopamine in the brain (Quinn, NP 1990. Levodopa-based therapy. Pages 169-184, in Therapy of Parkinson's disease. Marcel Dekker, New York). This pharmacological treatment is initially effective, but treatment with L-DOPA has serious side effects, such as the production of uncontrolled movements and dystonia (Kanazawa, I. 1986. Clinical pathophysilogy of basal ganglia diseases. VoI. 49, pages 65-92, in Vinken, PT, Bruyn, GW, Klawans, HL (eds): Handbook of Clinical Neurology. Elsevier Science Publishers, New York). So far there is no clear and effective cure for Parkinson's disease. Now the applicant has found that an extract from Ginkgo biloba also has new pharmacological properties, slowing and attenuating the dopaminergic damage produced in Parkinson's disease.

En particular el aplicante fue capaz de notar los efectos benéficos de un extracto de Ginkgo biloba en un modelo farmacológico de Ia enfermedad, utilizando Ia neurotoxina 1-metil-4-fenil-1 ,2,3,6-tetrahidropiridina denominada de aquí en adelante como MPTP.In particular, the applicant was able to notice the beneficial effects of an extract of Ginkgo biloba in a pharmacological model of the disease, using the neurotoxin 1-methyl-4-phenyl-1, 2,3,6-tetrahydropyridine named hereinafter like MPTP.

La enfermedad de Parkinson tiene una incidencia de 168/100,000 y es una enfermedad bien conocida especialmente en Ia población de una edad mayor de 60 años. Los síntomas de esta enfermedad incluyen los movimientos incontrolados, temblor en reposo, disturbios de Ia marcha, anormalidades posturales y rigidez muscular.Parkinson's disease has an incidence of 168 / 100,000 and is a well-known disease especially in the population over the age of 60 years. Symptoms of this disease include uncontrolled movements, tremor at rest, gait disturbances, postural abnormalities, and muscle stiffness.

La neuropatología primaria asociada con éste desorden es Ia pérdida progresiva y persistente de las neuronas dopaminérgicas originadas en Ia sustancia nigra pars compacta que proyectan al cuerpo estriado. Esto conduce a un decremento sustancial del neurotransmisor dopamina en Ia vía nigroestriatal (Jenner, P. 1989. Clues to the mechanism underlying dopamine cell death in Parkinson's disease. J. Neurol. Neurosurg. Psychiatry. (special suppl):22-28). El subsecuente decremento en Ia síntesis de dopamina correlaciona con el inicio y severidad de los síntomas mencionados. Es importante enfatizar que Ia síntesis de dopamina esta determinada por Ia disposición de Ia actividad de Ia tirosina hidroxilasa para llevar a cabo Ia síntesis de catecolaminas incluyendo Ia dopamina.The primary neuropathology associated with this disorder is the progressive and persistent loss of dopaminergic neurons originating from the nigra pars compact substance that project to the striatum. This leads to a substantial decrease in the neurotransmitter dopamine in the nigrostriatal pathway (Jenner, P. 1989. Clues to the mechanism underlying dopamine cell death in Parkinson's disease. J. Neurol. Neurosurg. Psychiatry. (Special suppl): 22-28). The subsequent decrease in the synthesis of dopamine correlates with the onset and severity of the mentioned symptoms. It is important to emphasize that the synthesis of dopamine is determined by the disposition of the activity of the tyrosine hydroxylase to carry out the synthesis of catecholamines including dopamine.

Aún cuando estos cambios han sido extensamente documentados, no ha sido identificada Ia causa que origina el proceso de degeneración neuronal. Sin embargo, hay evidencia creciente del papel que desempeña el proceso de estrés oxidativo producido por radicales libres en esta enfermedad. Los radicales libres son moléculas altamente reactivas producidas en el cuerpo como productos normales del metabolismo y en grandes cantidades por Ia inflamación, alcoholismo, fumar, radiación, etc. Esas moléculas altamente reactivas pueden oxidar otras moléculas en el cuerpo, haciéndolas inestables y potencialmente muy dañinas. Los radicales libres son moléculas altamente inestables que pueden causar daño al ácido desoxiribonucléico, carbohidratos, lípidos y proteínas en el cuerpo (Halliwell, B. and Gutteridge, J. M. C. Free radicáis in biology and medicine. Oxford-Claredon, 1985). De estos efectos, uno de los más importantes es Ia peroxidación de lípidos, el índice más comúnmente usado para medir el efecto biológico de los radicales libres. Este daño conocido como estrés oxidativo, está implicado en Ia patogénesis de un gran número de enfermedades crónicas incluyendo Ia enfermedad de Parkinson.Even though these changes have been extensively documented, the cause that originates the process of neuronal degeneration has not been identified. However, there is increasing evidence of the role that the oxidative stress process produced by free radicals plays in this disease. Free radicals are highly reactive molecules produced in the body as normal products of metabolism and in large quantities by Ia inflammation, alcoholism, smoking, radiation, etc. Those highly reactive molecules can oxidize other molecules in the body, making them unstable and potentially very harmful. Free radicals are highly unstable molecules that can cause damage to deoxyribonucleic acid, carbohydrates, lipids, and proteins in the body (Halliwell, B. and Gutteridge, JMC Free you are in biology and medicine. Oxford-Claredon, 1985). Of these effects, one of the most important is lipid peroxidation, the index most commonly used to measure the biological effect of free radicals. This damage known as oxidative stress, is implicated in the pathogenesis of a large number of chronic diseases, including Parkinson's disease.

Los organismos han desarrollado varios mecanismos de defensa para protegerse contra el daño de los radicales libres. Esos mecanismos de defensa incluyen las enzimas antioxidantes, atrapadores de los radicales libres, y agentes quelantes de metales (Reiter, RT. 1995. Oxidative processes and antioxidative defense mechanisms in the aging brain. FASEB J. 9: 526-533). Las enzimas antioxidantes son Ia catalasa, glutatión peroxidasa, y superóxido dismutasa. Normalmente, hay un balance entre Ia generación de los radicales libres y sistemas de defensa antioxidante in vivo. Cuando este balance es alterado a favor de Ia producción de radicales libres debido a Ia disminución de los sistemas antioxidantes o incremento en Ia generación de radicales libres, ocurre el estrés oxidativo.Organisms have developed various defense mechanisms to protect themselves against free radical damage. Those defense mechanisms include antioxidant enzymes, free radical scavengers, and metal chelating agents (Reiter, RT. 1995. Oxidative processes and antioxidative defense mechanisms in the aging brain. FASEB J. 9: 526-533). The antioxidant enzymes are catalase, glutathione peroxidase, and superoxide dismutase. Normally, there is a balance between the generation of free radicals and antioxidant defense systems in vivo. When this balance is altered in favor of the production of free radicals due to the decrease in antioxidant systems or an increase in the generation of free radicals, oxidative stress occurs.

El estrés oxidativo conduce al daño de los ácidos grasos poliinsaturados por peroxidación de lípidos, una reacción en cadena que resulta en numerosos productos de degradación (Niké, E., Noguchi, N. and Gotoh, N. 1993. Dynamics of lipid peroxidation and its inhibition by antioxidants. Biochem. Soc. Trans. 21: 313-317).Oxidative stress leads to damage to polyunsaturated fatty acids by lipid peroxidation, a chain reaction that results in numerous degradation products (Niké, E., Noguchi, N. and Gotoh, N. 1993. Dynamics of lipid peroxidation and its inhibition by antioxidants. Biochem. Soc. Trans. 21: 313-317).

Los estudios postmortem en pacientes con Ia enfermedad de Parkinson han reportado alteraciones relacionadas al estrés oxidativo en Ia sustancia nigra: el nivel del fierro está elevado (Youdim, M. B. H., Schachar, D. Ben., and Riederer, P. 1989. Is Parkinson's disease a progressive siderosis of substantia nigra resulting in increased ¡ron and melanin induced neurodegeneration? Acta Neurol. Scand. 126: 47-54), hay una pérdida del glutatión (Sian, J., Dexter, D.T., Lees, AJ. , Daniel, S., Agid, Y., Javoy-Agid, F., Jenner, P.P. and Marsden, CD. 1994. Alteration in glutathione levéis in Parkinson's disease and other neurodegenerative disorders affecting the basal ganglia. Ann. Neurol. 36: 348- 355), e incremento de Ia peroxidación de lípidos (Dexter, D.T., Cárter, CJ. , Wells, F.R., Javoy-Agid, F., Agid, Y., Lees, A., Jenner, P. and Marsden, CD. 1989. Basal lipid peroxidation in substantia nigra is increased in Parkinson's disease. J. Neurochem. 52: 381-389). De esta forma es de considerable interés Ia investigación de nuevas modalidades terapéuticas de Ia enfermedad.Postmortem studies in patients with Parkinson's disease have reported alterations related to oxidative stress in the substantia nigra: the level of iron is elevated (Youdim, MBH, Schachar, D. Ben., And Riederer, P. 1989. Is Parkinson's disease a progressive siderosis of substantia nigra resulting in increased ron and melanin induced neurodegeneration? Neurol. Scand. 126: 47-54), there is a loss of glutathione (Sian, J., Dexter, DT, Lees, AJ., Daniel, S., Agid, Y., Javoy-Agid, F., Jenner, PP and Marsden, CD. 1994. Alteration in glutathione levéis in Parkinson's disease and other neurodegenerative disorders affecting the basal ganglia. Ann. Neurol. 36: 348-355), and increased lipid peroxidation (Dexter, DT, Carter, CJ., Wells, FR, Javoy-Agid, F., Agid, Y., Lees, A., Jenner, P. and Marsden, CD. 1989. Basal lipid peroxidation in substantia nigra is increased in Parkinson's disease. J. Neurochem. 52: 381- 389). Thus, the investigation of new therapeutic modalities of the disease is of considerable interest.

La eficacia terapéutica de nuevos fármacos antiparkinsónicos es generalmente probado en modelos experimentales de Ia enfermedad. Así, Ia MPTP es referido como el mejor modelo experimental de Ia enfermedad de Parkinson (Gerlach, M., Riederer, P., Przuntek, H., and Youdim, M.B.H. 1991. MPTP mechanisms of neurotoxicity and their implications for Parkinson's disease. Eur. J. Pharmacol. Mol. Pharmacol. 208:273-286). Cuando Ia MPTP es administrada a primates que no son humanos y ratones produce hipokinesia y daño neuronal similar al observado en Ia enfermedad de Parkinson (Heikkila RE, Cabbat FS, Manzino L, Duvoisin RC 1984. Effects of MPTP on nigrostriatal dopamine in mice. Neuropharmacology 23: 711-713).The therapeutic efficacy of new antiparkinsonic drugs is generally tested in experimental models of the disease. Thus, MPTP is referred to as the best experimental model of Parkinson's disease (Gerlach, M., Riederer, P., Przuntek, H., and Youdim, MBH 1991. MPTP mechanisms of neurotoxicity and their implications for Parkinson's disease. Eur J. Pharmacol. Mol. Pharmacol. 208: 273-286). When MPTP is administered to primates that are not human and mice, it produces hypokinesia and neuronal damage similar to that observed in Parkinson's disease (Heikkila RE, Cabbat FS, Manzino L, Duvoisin RC 1984. Effects of MPTP on nigrostriatal dopamine in mice. Neuropharmacology 23: 711-713).

El ion 1 -metil-4-fenilpiridino que denominaremos de aquí en adelante como MPP+ es el mayor metabolito neurotóxico de Ia MPTP y es el responsable de sus efectos neurotóxicos. La biotransformación de Ia MPTP a su metabolito tóxico MPP+ en el cerebro es inicialmente realizada por Ia enzima monoamino- oxidasa B que denominaremos de aquí en adelante como MAO-B y esta genera radicales libres (Zang, L.Y. and Misra, H. P. 1993. Generation of reactive oxygen species during the monoamine oxidase-catalyzed oxidation of the neurotoxicant, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine. J. Biol. Chem. 268:16504-16512). La monoamino-oxidasa que denominaremos de aquí en adelante comoThe 1-methyl-4-phenylpyridine ion that we will hereinafter refer to as MPP + is the largest neurotoxic metabolite of MPTP and is responsible for its neurotoxic effects. The biotransformation of the MPTP to its toxic metabolite MPP + in the brain is initially carried out by the monoamine oxidase B enzyme that we will hereinafter refer to as MAO-B and this generates free radicals (Zang, LY and Misra, HP 1993. Generation of reactive oxygen species during the monoamine oxidase-catalyzed oxidation of the neurotoxicant, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine. J. Biol. Chem. 268: 16504-16512). The monoamine oxidase that we will hereinafter refer to as

MAO existe en 2 isoformas A y B. La MAO es Ia mayor enzima intracelular que metaboliza Ia dopamina en el sistema nervioso central. La oxidación elevada de Ia dopamina por Ia MAO-B en Ia enfermedad de Parkinson puede involucrar Ia producción de peróxido de hidrógeno, una molécula tóxica derivada del oxígeno capaz de inducir Ia formación de radicales libres, Ia cual podría potencialmente dañar a las neuronas dopaminérgicas de Ia vía nigroestriatal (Cohén, G. 1986. Monoamine oxidase, hydrogen peroxide, and Parkinson's disease. Pages 119- 125, in Yahr, M.D., and Bergmann, KJ. , (eds.), Advances in Neurplogy: Parkinson's disease, vol. 45, Raven Press, New York).MAO exists in 2 isoforms A and B. MAO is the largest intracellular enzyme that metabolizes dopamine in the central nervous system. The high oxidation of dopamine by MAO-B in Parkinson's disease may involve Ia production of hydrogen peroxide, a toxic molecule derived from oxygen capable of inducing the formation of free radicals, which could potentially harm dopaminergic neurons of the nigrostriatal pathway (Cohén, G. 1986. Monoamine oxidase, hydrogen peroxide, and Parkinson's disease . Pages 119- 125, in Yahr, MD, and Bergmann KJ (eds.), Advances in Neurplogy. Parkinson 's disease, vol 45, Raven Press, New York)..

Los pacientes con Ia enfermedad de Parkinson tienen una elevada actividad de Ia enzima MAO-B en Ia sustancia nigra e inhibidores de Ia MAO-B tienen efecto potencial neuroprotector en esta enfermedad (Knoll, J. 1995. Rationale for (-) deprenyl (selegiline) medication in Parkinson's disease and in prevention of age-related nigral changes. Biomed. Pharmacother. 49:187-195) y en Ia neurotoxicidad de Ia MPTP/MPP+ (Knoll, J. 1995. Rationale for (-) deprenyl (selegiline) medication in Parkinson's disease and in prevention of age-related nigral changes. Biomed. Pharmacother. 49:187-195; Wu, R. M., Mohannakumar, K.P., Murphy, D. L., and Chiueh, C. C. 1994. Antioxidant mechanism and protection of nigral neurons against MPP+ toxicity by deprenyl (selegiline). Ann. N.Y. Acad. Sci. 738:214-221).Patients with Parkinson's disease have a high activity of the MAO-B enzyme in the nigra and MAO-B inhibitors have a potential neuroprotective effect in this disease (Knoll, J. 1995. Rationale for (-) deprenyl (selegiline ) medication in Parkinson's disease and in prevention of age-related nigral changes. Biomed. Pharmacother. 49: 187-195) and in the neurotoxicity of MPTP / MPP + (Knoll, J. 1995. Rationale for (-) deprenyl (selegiline) medication in Parkinson 's disease and in prevention of age-related changes Biomed Pharmacother nigral 49: 187-195; Wu, RM, Mohannakumar, KP, Murphy, DL, and Chiueh, CC 1994. Antioxidant protection of nigral mechanism and... neurons against MPP + toxicity by deprenyl (selegiline). Ann. NY Acad. Sci. 738: 214-221).

Como se mencionó, en Ia actualidad Ia terapia de elección para pacientes que tienen enfermedad de Parkinson es a través de Ia administración de Ia L-DOPA, que es convertida a dopamina en el cerebro. Este tratamiento farmacológico es inicialmente efectivo, pero con frecuencia llega a ser menos efectivo sobre el tiempo y en muchos casos los síntomas de los pacientes llegan a empeorar. Hasta el momento no hay clara y efectiva cura para Ia enfermedad de Parkinson.As mentioned, currently the therapy of choice for patients who have Parkinson's disease is through the administration of L-DOPA, which is converted to dopamine in the brain. This pharmacological treatment is initially effective, but it often becomes less effective over time and in many cases the symptoms of the patients get worse. So far there is no clear and effective cure for Parkinson's disease.

La presente invención representa una alternativa potencial e importante para Ia terapia usada para el tratamiento de Ia enfermedad de Parkinson. Esto es anticipado que el tratamiento con un extracto del Ginkgo biloba a pacientes con Ia enfermedad de Parkinson inhibirá ó detendrá el progreso de Ia enfermedad por reducir Ia degeneración y disfunción de las neuronas dopaminérgicas de Ia vía nigroestriatal. Así mismo, actuar como un agente antioxidante y/o antilipoperoxidante, regular y/o inhibir Ia actividad de Ia MAO que es generadora de radicales libres; e incrementar Ia actividad de Ia tirosina hidroxilasa, enzima requerida para Ia síntesis de dopamina.The present invention represents a potential and important alternative for the therapy used for the treatment of Parkinson's disease. This is anticipated that treatment with an extract of Ginkgo biloba to patients with Parkinson's disease will inhibit or stop the progress of the disease by reducing the degeneration and dysfunction of dopaminergic neurons of the nigrostriatal pathway. Likewise, act as an antioxidant and / or antilipoperoxidant agent, regulate and / or inhibit the activity of the MAO that is generating free radicals; and increase the activity of tyrosine hydroxylase, enzyme required for the synthesis of dopamine.

Por extracto de Ginkgo biloba se entiende que es al menos un extracto de los compuestos individuales que pueden ser obtenidos por extracción del árbol Ginkgo biloba y en particular un compuesto flavonoide o un terpeno como un ginkgólido o un bilobálido, o también una mezcla. Para los usos de acuerdo a Ia invención, un extracto de tipo EGb 761 puede por ejemplo ser escogido. El extracto del EGb 761 es una mezcla bien definida de compuestos activos extraídos de las hojas del Ginkgo biloba. Este es un extracto de las hojas de Ginkgo biloba el cual es sustancialmente libre de compuestos alquifenoles, tiene un alto contenido de glicósidos de flavonoides, y el cual contiene substancialmente todos los ginkgólidos y bilobálidos originalmente presentes en las hojas.By Ginkgo biloba extract is meant that it is at least an extract of the individual compounds that can be obtained by extraction from the Ginkgo biloba tree and in particular a flavonoid compound or a terpene such as a ginkgolide or a bilobalide, or also a mixture. For the uses according to the invention, an extract of the EGb 761 type can for example be chosen. EGb 761 extract is a well-defined mixture of active compounds extracted from the leaves of Ginkgo biloba. This is an extract from the leaves of Ginkgo biloba which is substantially free of alkyphenol compounds, has a high content of flavonoid glycosides, and which contains substantially all of the ginkgolides and bilobalides originally present in the leaves.

Por extracto de tipo EGb 761 se entiende a un extracto de una composición substancialmente idéntica al del extracto estandarizado EGb 761 definido en el siguiente artículo: K. Drieu, La presse medícale, 31 , Sep. 25,By extract of the EGb 761 type is meant an extract of a composition substantially identical to that of the standardized extract EGb 761 defined in the following article: K. Drieu, La presse medícale, 31, Sep. 25,

1986, supplement devote to the extract of Ginkgo biloba (EGb 761), 1455-1457; o en las patentes Europeas EP 431 535 y EP 431 536; y Ia patente americana1986, supplement devote to the extract of Ginkgo biloba (EGb 761), 1455-1457; or in the European patents EP 431 535 and EP 431 536; and the American patent

US Pat. 5,399,348. Por extracto de tipo EGb 761 es por Io tanto entendido en particular de extractos de Ginkgo biloba comprendiendo 20 a 30% glicósidos de flavonas, 2.5 a 4.5% de ginkgólidos A, B, C, y J, 2 a 4% de bilobálidos, menos del 10% de proantocianidinas y menos de 10 ppm, y preferiblemente menos de 5 ppm, de compuestos de tipo alquifenol, y en particular extractos de Ginkgo biloba comprendiendo aproximadamente 24% glicósidos de flavonas, 3.1% de ginkgólidos A, B, C y J, 2.9% de bilobálidos, 6.5% de proantocianidinas y menos de 1 ppm de compuestos de tipo alquifenol.US Pat. 5,399,348. By extract of type EGb 761 is by Io understood in particular of extracts of Ginkgo biloba comprising 20 to 30% glycosides of flavones, 2.5 to 4.5% of ginkgolides A, B, C, and J, 2 to 4% of bilobalides, less 10% proanthocyanidins and less than 10 ppm, and preferably less than 5 ppm, of compounds of the alkylphenol type, and in particular extracts of Ginkgo biloba comprising approximately 24% flavone glycosides, 3.1% ginkgolides A, B, C and J , 2.9% of bilobalides, 6.5% of proanthocyanidins and less than 1 ppm of compounds of the alkylphenol type.

El EGb 761 ha sido usado en Ia clínica para el tratamiento de Ia insuficiencia cerebrovascular, demencia degenerativa, alteraciones vasculares periféricas y desórdenes neurosensoriales (DeFeudis, F.V.1998. Gingko biloba extract (EGb761): from Chemistry to the clinic. Pages 401. Ullstein Medical,EGb 761 has been used in the clinic for the treatment of cerebrovascular insufficiency, degenerative dementia, peripheral vascular disorders and sensorineural disorders (DeFeudis, FV1998. Gingko biloba extract (EGb761): from Chemistry to the clinic. Pages 401. Ullstein Medical ,

Wiesbaden).Wiesbaden).

Una acción polivalente del EGb 761 es responsable de su eficacia en tratar desórdenes clínicos de origen multifactorial. Las subfracciones de varios constituyentes químicos del EGb 761, aunque activos en modelos farmacológicos, no generalmente reproduce las acciones del extracto total. Así los efectos aditivos, antagonistas y sinergistas de los constituyentes activos del EGb 761 probablemente ocurre con respecto a diversos sitios blanco moleculares en diferentes tejidos y órganos.A multipurpose action of EGb 761 is responsible for its effectiveness in treat clinical disorders of multifactorial origin. The subfractions of various chemical constituents of EGb 761, although active in pharmacological models, do not generally reproduce the actions of the total extract. Thus the additive, antagonistic and synergistic effects of the active constituents of EGb 761 probably occur with respect to various molecular target sites in different tissues and organs.

Ha sido propuesto que los efectos benéficos del EGb 761 en el sistema nervioso central, son probablemente por su acción antioxidante, por atrapar los radicales superóxido, hidroxilo, y peroxil (Marcocci, L., Packer, L., Droy-Lefaix, M., Sekaki, A., and Gardes-Albert, M. 1994. Antioxidant action of Ginkgo biloba extract EGb 761. Methods Enzymol.. 234: 462-475), interactuar con el óxido nítrico (Marcocci, L., Maguire, J. J., Droy-Lefaix, M.T., Packer, L. 1994. The nitric oxide-scavenging properties of Ginkgo biloba extract EGb 761. Biochem. Biophys. Res. Común. 201: 748-755) inhibir y/o regular Ia actividad de Ia MAO que produce radicales libres (DeFeudis, F. V. 1998. Ginkgo biloba extract (EGb761): from Chemistry to the clinic. Pages 401. Ullstein Medical, Wiesbaden).It has been proposed that the beneficial effects of EGb 761 on the central nervous system are probably due to its antioxidant action, because it traps superoxide, hydroxyl, and peroxyl radicals (Marcocci, L., Packer, L., Droy-Lefaix, M. , Sekaki, A., and Gardes-Albert, M. 1994. Antioxidant action of Ginkgo biloba extract EGb 761. Methods Enzymol .. 234: 462-475), interact with nitric oxide (Marcocci, L., Maguire, JJ, Droy-Lefaix, MT, Packer, L. 1994. The nitric oxide-scavenging properties of Ginkgo biloba extract EGb 761. Biochem. Biophys. Common Res. 201: 748-755) inhibit and / or regulate the activity of MAO that produces free radicals (DeFeudis, FV 1998. Ginkgo biloba extract (EGb761): from Chemistry to the clinic. Pages 401. Ullstein Medical, Wiesbaden).

Finalmente el objeto de esta invención es proveer fármacos que contienen este extracto de Ginkgo biloba con un alto contenido de glicósidos de flavonoides, ginkgólidos y bilobálidos originalmente presentes en las hojas y donde no hay sustancialmente peligro de reacciones alérgicas, precisamente por Ia remoción de los compuestos alquifenoles. Preferiblemente el extracto en Ia presente invención debe contener: - de 20 a 30 porcentaje en peso, en particular 22 a 26 de porcentaje en peso de glicósidos de flavonoides.Finally, the object of this invention is to provide drugs that contain this extract of Ginkgo biloba with a high content of flavonoid glycosides, ginkgolides and bilobalides originally present in the leaves and where there is no substantial danger of allergic reactions, precisely due to the removal of the compounds alkyphenols. Preferably the extract in the present invention should contain: - from 20 to 30 percent by weight, in particular 22 to 26 percent by weight of flavonoid glycosides.

-2.5 a 4.5 de porcentaje en peso de ginkgólidos A, B, C y J en total. -2.0 a 4.0 de porcentaje en peso de bilobálidos,-2.5 to 4.5% by weight of ginkgolides A, B, C and J in total. -2.0 to 4.0 of percentage by weight of bilobalides,

-Menos de 10 ppm, en particular menos de 1 ppm de compuestos alquifenoles y-Less than 10 ppm, in particular less than 1 ppm of alkyphenol compounds and

-Menos de 10 de porcentaje en peso de proantocianidinas.-Less than 10 weight percent of proanthocyanidins.

El extracto de Ginkgo biloba de Ia invención puede ser procesado en Ia vía usual de preparaciones farmacéuticas, por ejemplo para soluciones, tabletas cubiertas, preparaciones de tabletas o inyecciones.The Ginkgo biloba extract of the invention can be processed in Ia usual route of pharmaceutical preparations, for example for solutions, coated tablets, tablet preparations or injections.

Ahora el aplicante ha encontrado que cierto extracto de Ginkgo biloba tiene un uso diferente debido a sus nuevas propiedades, como es retardar y atenuar el daño dopaminérgico producido durante Ia enfermedad de Parkinson.Now the applicant has found that a certain extract of Ginkgo biloba has a different use due to its new properties, such as retarding and attenuating the dopaminergic damage produced during Parkinson's disease.

Así mismo, actuar como un agente antioxidante y/o antilipoperoxidante, regular y/o inhibir Ia actividad de Ia MAO que es generadora de radicales libres; incrementar Ia actividad de Ia tirosina hidroxilasa enzima requerida para Ia síntesis de dopamina.Likewise, act as an antioxidant and / or antilipoperoxidant agent, regulate and / or inhibit the activity of the MAO that is generating free radicals; increase the activity of the tyrosine hydroxylase enzyme required for the synthesis of dopamine.

Por Io tanto el objeto de Ia presente invención es el uso de un extracto de Ginkgo biloba, para preparar un medicamento destinado para tratar Ia enfermedad de Parkinson.Therefore, the object of the present invention is the use of an extract of Ginkgo biloba, to prepare a medicine intended to treat Parkinson's disease.

Los compuestos pueden ser administrados varios meses para el tratamiento. Las composiciones farmacéuticas comprenden un compuesto de Ia invención que puede estar en forma de sólidos, por ejemplo polvos, tabletas, cápsulas, liposomas o supositorios. Los excipientes pueden ser por ejemplo fosfato de calcio, estearato de magnesio, talco, azúcares, lactosa, dextrina, gelatina, almidón, celulosa, metil celulosa, celulosa carboximetil de sodio, polivinilpirrolidina y cera.The compounds can be administered for several months for treatment. The pharmaceutical compositions comprise a compound of the invention that can be in the form of solids, for example powders, tablets, capsules, liposomes or suppositories. The excipients can be for example calcium phosphate, magnesium stearate, talc, sugars, lactose, dextrin, gelatin, starch, cellulose, methyl cellulose, sodium carboxymethyl cellulose, polyvinylpyrrolidine and wax.

Las composiciones farmacéuticas conteniendo un compuesto de Ia invención puede también estar presente en Ia forma de un líquido, por ejemplo, soluciones, emulsiones, suspensiones o jarabe. Los excipientes apropiados para los líquidos pueden ser por ejemplo, agua, solventes orgánicos como el glicerol ó los glicoles, así como sus mezclas, en varias proporciones.The pharmaceutical compositions containing a compound of the invention may also be present in the form of a liquid, for example, solutions, emulsions, suspensions or syrup. Suitable excipients for liquids can be, for example, water, organic solvents such as glycerol or glycols, as well as their mixtures, in various proportions.

La administración del medicamento de acuerdo a Ia invención puede ser tópico, oral, ruta parenteral, por inyección, como Ia administración intramuscular, subcutánea, intravenosa, etc.The administration of the medicine according to the invention can be topical, oral, parenteral route, by injection, such as intramuscular, subcutaneous, intravenous administration, etc.

La dosis de administración diaria de los ingredientes activos que pueden ser usados de acuerdo a Ia invención esta comprendido entre 0.1 mg a 1 g dependiendo de Ia gravedad de Ia enfermedad de Parkinson del paciente que será tratado. Preferiblemente, para los extractos de Ginkgo biloba, una dosis que comprenda entre 5 mg a 600 mg por día aproximadamente puede ser escogida.The daily administration dose of the active ingredients that can be used according to the invention is between 0.1 mg to 1 g depending on the severity of the Parkinson's disease of the patient to be treated. Preferably, for Ginkgo biloba extracts, a dose ranging from approximately 5 mg to 600 mg per day may be chosen.

Los siguientes ejemplos son sólo ilustrativos y no pretenden limitar Ia invención en ninguna forma. Utilize un modelo experimental de Ia enfermedad de Parkinson involucrado en Ia generación de los radicales hidroxilo como es Ia MPTP, antes mencionada (Chiueh CC1 Huang SJ, and Murphy DL. 1992. Enhanced hydroxyl radical generation by 2'-methyl analog of MPTP: suppression by clorgyline and deprenyl. Synapse. 11(4):346-8). Esto fue con el propósito de evaluar el efecto protector del EGb 761 en el daño inducido por radicales libres, así como su efecto restaurador en los niveles de dopamina. La neurotoxina MPTP ha mostrado conducir Ia degeneración de las neuronas dopaminérgicas en el cerebro, así proveyendo un buen modelo de Ia enfermedad de Parkinson inducida experimentalmente. Este modelo es ahora ampliamente aceptado y es usado para evaluar agentes potenciales en Ia terapia para esta enfermedad.The following examples are only illustrative and are not intended to limit the invention in any way. Use an experimental model of Parkinson's disease involved in the generation of hydroxyl radicals such as MPTP, mentioned above (Chiueh CC 1 Huang SJ, and Murphy DL. 1992. Enhanced hydroxyl radical generation by 2'-methyl analog of MPTP: suppression by clorgyline and deprenyl. Synapse. 11 (4): 346-8). This was for the purpose of evaluating the protective effect of EGb 761 on free radical-induced damage, as well as its restorative effect on dopamine levels. The MPTP neurotoxin has been shown to drive the degeneration of dopaminergic neurons in the brain, thus providing a good model of experimentally induced Parkinson's disease. This model is now widely accepted and is used to evaluate potential agents in therapy for this disease.

Esos datos demuestran que este particular extracto del Ginkgo biloha desplaza alta eficacia terapéutica in vivo en un modelo de roedores de Ia enfermedad en el humano, y también indica que este extracto cruza Ia barrera hematoencefálica para prevenir el daño inducido por los radicales libres por MPTP.These data demonstrate that this particular Ginkgo biloha extract displaces high therapeutic efficacy in vivo in a rodent model of the disease in humans, and also indicates that this extract crosses the blood-brain barrier to prevent damage caused by free radicals by MPTP.

Breve descripción de los dibujosBrief description of the drawings

Para un mejor entendimiento de Ia invención se presenta ésta con Ia asistencia de 2 dibujos:For a better understanding of the invention, it is presented with the assistance of 2 drawings:

Figura 1. Esta gráfica muestra que el tratamiento con un extracto de Ginkgo biloba tipo EGb 761 regula Ia actividad de Ia MAO B en el cuerpo estriado en el parkinsonismo inducido con el MPP+. Los resultados son expresados en μmoles de 4 HOQV/g/h. Cada barra representa el promedio +/- el error estándar de 4-6 experimentos por grupo. *Estadisticamente diferente del grupo control, representado como "salina + salina", p<0.05, prueba de Tukey. +Estadisticamente diferente del grupo de MPP+, representado como "salina + MPP+", p<0.05, prueba de Tukey. EGb 761= extracto de Ginkgo biloba tipo EGb 761; MPP+ = ion 1-met¡l-4-fenilpiridino; 4-HOQ = 4-h¡droxiquinolina.Figure 1. This graph shows that treatment with an extract of Ginkgo biloba type EGb 761 regulates the activity of MAO B in the striatum in parkinsonism induced with MPP + . The results are expressed in µmoles of 4 HOQV / g / h. Each bar represents the average +/- standard error of 4-6 experiments per group. * Statistically different from the control group, represented as "saline + saline", p <0.05, Tukey's test. + Statistically different from the MPP + group , represented as "saline + MPP + ", p <0.05, Tukey test. EGb 761 = Ginkgo biloba extract type EGb 761; MPP + = 1-methyl-4-phenylpyridine ion; 4-HOQ = 4-hydroxyquinoline.

Figura 2. Es una gráfica que muestra Ia restauración que produce el extracto de Ginkgo biloba tipo EGb 761 en los niveles de dopamina en el cuerpo estriado en animales que fueron parkinsonizados previamente con MPTP. Donde A representa el grupo de " salina + salina ", B representa el grupo de " salina + EGb 761 a una dosis de 40 mg/kg ", C representa el grupo de " salina + EGb 761 a una dosis de 120 mg/kg ", D representa el grupo de "MPTP + salina", E representa el grupo de "MPTP + EGb 761 a una dosis de 40 mg/kg", F representa el grupo de "MPTP + EGb 761 a una dosis de 120 mg/kg". Los resultados son expresados en contenido de dopamina en μg/g de tejido. Cada barra representa el promedio ± error estándar de 6-8 experimentos por grupo. * Estadísticamente diferente del grupo tratado con MPTP representado como " salina + MPTP ", p<0.05, prueba de Tukey. **Estadísticamente diferente del grupo tratado con MPTP representado por "salina + MPTP", p<0.01 , prueba de Tukey. +Estadíst¡camente diferente del grupo control representado por "salina + salina", p<0.05, prueba de Tukey. EGb 761= extracto de Ginkgo biloba tipo EGb 761 ; MPTP = 1-metil-4-fenil-1 , 2, 3, 6-tetrahidropiridina.Figure 2. It is a graph that shows the restoration that produces the extract of Ginkgo biloba type EGb 761 in the levels of dopamine in the striatum in animals that were previously parkinsonized with MPTP. Where A represents the "saline + saline" group, B represents the "saline + EGb 761 group at a dose of 40 mg / kg", C represents the "saline + EGb 761 group at a dose of 120 mg / kg ", D represents the" MPTP + saline "group, E represents the" MPTP + EGb 761 group at a dose of 40 mg / kg ", F represents the" MPTP + EGb 761 group at a dose of 120 mg / kg ". The results are expressed in dopamine content in µg / g of tissue. Each bar represents the mean ± standard error of 6-8 experiments per group. * Statistically different from the group treated with MPTP represented as "saline + MPTP", p <0.05, Tukey's test. ** Statistically different from the group treated with MPTP represented by "salina + MPTP", p <0.01, Tukey's test. + Statistically different from the control group represented by "saline + saline", p <0.05, Tukey's test. EGb 761 = Ginkgo biloba extract type EGb 761; MPTP = 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine.

Los siguientes ejemplos son sólo ilustrativos y no para limitar Ia invención en ninguna fase. Los equipos utilizados no es una limitante, porque equipo equivalente, alternativo o similar se puede usar para desarrollar Ia presente invención. Para mostrar Ia importancia del uso de un extracto de Ginkgo biloba tipo EGb 761 en el tratamiento de Ia enfermedad de Parkinson, se llevaron a cabo las siguientes pruebas:The following examples are only illustrative and not to limit the invention in any phase. The equipment used is not a limitation, because equivalent, alternative or similar equipment can be used to develop the present invention. To show the importance of the use of an extract of Ginkgo biloba type EGb 761 in the treatment of Parkinson's disease, the following tests were carried out:

Los ejemplos del 3 al 7 fueron llevados a cabo utilizando ratones macho de Ia cepa C-57 black de 25-30 g de peso. Los animales fueron mantenidos en condiciones estándar con un ciclo de 12:12h luz /oscuridad, 21-22 °C, humedad relativa 40%, con alimento y agua ad libitum. El extracto de Ginkgo biloba tipo EGb 761 fue donado generosamente por Dr. Willmar Schwabe GmbH & Co. KG en Karlsruhe, Alemania. El EGb 761 fue disuelto en solución salina y ajustado a un pH de 7.4.Examples 3 to 7 were carried out using male mice of the C-57 black strain of 25-30 g in weight. The animals were kept in standard conditions with a cycle of 12: 12h light / dark, 21-22 ° C, relative humidity 40%, with food and water ad libitum. Ginkgo biloba extract type EGb 761 was generously donated by Dr. Willmar Schwabe GmbH & Co. KG in Karlsruhe, Germany. EGb 761 was dissolved in saline and adjusted to a pH of 7.4.

Ejemplo 1Example 1

Solución para administración oral donde 10O mI de solución contienen:Solution for oral administration where 10O ml of solution contain:

extracto de Ginkgo biloba 4.0 gGinkgo biloba extract 4.0 g

Etanol 50.0 g Agua desmineralizada a 100.0 gEthanol 50.0 g Demineralized water at 100.0 g

Ejemplo 2Example 2

Tabletas cubiertas, donde una tableta contieneCovered tablets, where one tablet contains

extracto de Ginkgo biloba 40.00 mgGinkgo biloba extract 40.00 mg

Celulosa microcristalina 100.00 mgMicrocrystalline cellulose 100.00 mg

Lactosa 80.00 mg Ácido silícico coloidal 25.00 mgLactose 80.00 mg Colloidal silicic acid 25.00 mg

Talco en esencia 4.50 mgTalc in essence 4.50 mg

Estearato de magnesio 0.50 mgMagnesium stearate 0.50 mg

Metilcelulosa hidroxipropil 12.00 mgHydroxypropyl methylcellulose 12.00 mg

Pigmento de óxido férrico 0.10 mg Talco en recubrimiento 0.50 mgFerric oxide pigment 0.10 mg Talc in coating 0.50 mg

Peso de Ia tableta cubierta aproximadamente 262.60 mgWeight of the covered tablet approximately 262.60 mg

Ejemplo 3Example 3

En esta parte de Ia investigación encontré que el tratamiento con el extracto tipo EGb 761 actúa como un agente antioxidante/antiperoxidante en el parkinsonismo inducido con MPP+. Se diseñaron 4 grupos experimentales por grupo: grupo I: solución salina por vía intraperitoneal denominado de aquí en adelante como i.p + solución salina por vía intracerebroventricular denominado de aquí en adelante como icv; grupo II: EGb 761 por vía i.p + solución salina por vía icv; grupo III: solución salina por vía ip + MPP+ por vía icv; grupo IV: EGb 761 por vía i.p + MPP+ por vía icv.In this part of the investigation I found that the treatment with the EGb 761 type extract acts as an antioxidant / anti-oxidant agent in MPP + -induced parkinsonism. 4 experimental groups were designed for group: group I: saline intraperitoneally, hereinafter referred to as ip + saline intracerebroventricular, hereinafter referred to as icv; group II: EGb 761 by ip route + saline solution by icv route; group III: saline by ip route + MPP + by icv route; group IV: EGb 761 by ip route + MPP + by icv route.

Los animales de los grupos I y III recibieron solución salina por vía i.p y los grupos Il y IV recibieron extracto tipo EGb 761 a diferentes dosis que fueron 2.5, 5 o 10 mg/kg, i.p. durante 17 días. Después del tratamiento, los animales de los grupos III y IV fueron anestesiados con éter y administrados con 3 μl de una solución conteniendo 18 μg de MPP+ que corresponde a una dosis de 0.72 mg/kg, inyectados por vía icv como se describió previamente (Haley, TJ. and McCormick, W.G. 1957. Pharmacological effects produced by intracerebroventricular injection of drugs in the conscious mouse. Br. J. Pharmacol. 12(1): 12-15).Animals in groups I and III received saline ip and groups Il and IV received EGb 761 type extract at different doses that were 2.5, 5 or 10 mg / kg, ip for 17 days. After treatment, animals in groups III and IV were anesthetized with ether and administered with 3 µl of a solution containing 18 µg of MPP + corresponding to a dose of 0.72 mg / kg, injected via the icv route as previously described ( Haley, TJ. And McCormick, WG 1957. Pharmacological effects produced by intracerebroventricular injection of drugs in the conscious mouse. Br. J. Pharmacol. 12 (1): 12-15).

Los ratones de los grupos I y Il recibieron solución salina por vía icv y fueron el grupo control. Los ratones fueron sacrificados por dislocación cervical a 2 h después de Ia administración del MPP+. Los cerebros fueron inmediatamente removidos, el cuerpo estriado fue disectado y se analizó Ia actividad antioxidante/antiperoxidante evaluando Ia peroxidación de lípidos por medir los niveles de formación de productos fluorescentes lipidíeos que denominaremos de aquí en adelante como PFL usando una técnica previamente descrita (Triggs, WJ. and Willmore, LJ. 1984. In vivo lipid peroxidation in rat brain following intracortical Fe 2* injection. J. Neurochem. 42 (4): 976-980).Mice in groups I and Il received saline via the icv route and were the control group. Mice were sacrificed by cervical dislocation at 2 h after administration of MPP + . The brains were immediately removed, the striatum was dissected and the antioxidant / anti-oxidant activity was analyzed evaluating the lipid peroxidation by measuring the levels of formation of lipid fluorescent products that we will hereinafter refer to as PFL using a previously described technique (Triggs, WJ. And Willmore, LJ. 1984. In vivo lipid peroxidation in rat brain following intracortical Fe 2 * injection. J. Neurochem. 42 (4): 976-980).

Después de obtener el cuerpo estriado y pesar el tejido se homogenizó en 2.5 mi de buffer de fosfatos con un pH de 7.0. Se tomaron alícuotas de 1 mi del homogenado y se colocaron en tubos de vidrio con tapa. Posteriormente adicioné 3 mi de una mezcla de cloroformo-metanol a una dilución 2:1 volumen/volumen. Los tubos se taparon y Ia mezcla se agitó suavemente y se colocó en hielo durante 30 minutos. La fase acuosa fue descartada y 1 mi de Ia fase clorofórmica fue transferida en celdas de quarzo, posteriormente se adicionaron 0.1 mi de metanol. La fluorescencia fue medida en un espectrofotómetro para luminiscencia Perkin-Elmer LS 55 con una longitud de excitación de 370 nm y 430 nm de emisión. Previo a Ia medición de las muestras, Ia sensibilidad del espectrofotómetro de fluorescencia se ajustó a 140 unidades de fluorescencia con un estándar de quinina a una concentración de 0.1 μg/ml preparado en una solución de ácido sulfúrico a una concentración de 0.05 M. Los resultados se expresaron como unidades de fluorescencia/gramo de tejido húmedo/ml de extracto leído. Todas las muestras fueron analizadas en duplicado. En Ia tabla 1 se muestra que Ia administración de extracto tipo EGb 761 no produjo incrementos significantes en Ia formación de PFL a las diferentes dosis probadas que fueron 2.5, 5 y 10 mg/kg, a 2 h después de Ia administración de MPP+ comparado a los animales control. La peroxidación de lípidos a 2 h después de Ia administración de MPP+ sin pretratamiento con extracto tipo EGb 761 produjo un incremento en Ia peroxidación de lípidos de 72%. En contraste, como observamos en Ia tabla 1 el pretratamiento con extracto tipo EGb 761 a 10 mg/kg bloqueó en un 100% Ia peroxidación de lípidos inducida por el MPP+, es decir los niveles de Ia peroxidación de lípidos no son estadisticamente diferentes de los controles tratados con salina. En Ia tabla 1 se muestra el estudio dosis respuesta del extracto tipo EGbAfter obtaining the striatum and weighing, the tissue was homogenized in 2.5 ml of phosphate buffer with a pH of 7.0. 1 ml aliquots of the homogenate were taken and placed in glass tubes with lids. Subsequently I added 3 ml of a chloroform-methanol mixture at a 2: 1 volume / volume dilution. The tubes were capped and the mixture was gently shaken and placed on ice for 30 minutes. The aqueous phase was discarded and 1 ml of the chloroform phase was transferred in quartz cells, 0.1 ml of methanol were subsequently added. Fluorescence was measured on a Perkin-Elmer LS 55 luminescence spectrophotometer with an excitation length of 370nm and 430nm emission. Before measuring the samples, the sensitivity of the fluorescence spectrophotometer was adjusted to 140 fluorescence units with a quinine standard at a concentration of 0.1 μg / ml prepared in a solution of sulfuric acid at a concentration of 0.05 M. The results they were expressed as fluorescence units / gram of wet tissue / ml of read extract. All samples were analyzed in duplicate. Table 1 shows that the administration of extract type EGb 761 did not produce significant increases in the formation of PFL at the different doses tested, which were 2.5, 5 and 10 mg / kg, at 2 h after the administration of MPP + compared to control animals. The lipid peroxidation at 2 h after the administration of MPP + without pretreatment with EGb 761 type extract produced an increase in lipid peroxidation of 72%. In contrast, as we observed in Table 1, the pretreatment with extract type EGb 761 at 10 mg / kg blocked 100% the lipid peroxidation induced by MPP + , that is, the levels of lipid peroxidation are not statistically different from controls treated with saline. Table 1 shows the dose response study of the EGb type extract

761. El grado de protección incrementó con el incremento de Ia dosis del extracto tipo EGb 761. Por ejemplo, el grado de protección fue 65%, 93% y 100% para 2.5, 5 y 10 mg/kg del EGb 761 , respectivamente, cuando se compara con el grupo de "salina + MPP+".761. The degree of protection increased with increasing dose of the EGb 761 type extract. For example, the degree of protection was 65%, 93% and 100% for 2.5, 5 and 10 mg / kg of EGb 761, respectively, when compared to the "saline + MPP + " group.

Ejemplo 4Example 4

En esta invención encontré que el tratamiento con el extracto tipo EGb 761 regula Ia actividad de Ia enzima MAO en el parkinsonismo inducido con el MPP+. Se diseñaron 4 grupos experimentales y fueron tratados experimentalmente exactamente como se describió en el ejemplo 3. Posteriormente se analizó Ia actividad de Ia MAO. Después de Ia manipulación experimental los animales fueron sacrificados por dislocación cervical a 2, 6, 12 y 24 h después de Ia administración con MPP+. Los cerebros fueron TABLA 1. Peroxidación de lípidos en cuerpo estriadoIn this invention I found that the treatment with the EGb 761 type extract regulates the activity of the MAO enzyme in parkinsonism induced with MPP + . 4 experimental groups were designed and were experimentally treated exactly as described in Example 3. Subsequently, the activity of the MAO was analyzed. After the experimental manipulation the animals were sacrificed by cervical dislocation at 2, 6, 12 and 24 h after the administration with MPP + . The brains were TABLE 1. Lipid peroxidation in the striatum

Grupo experimental Peroxidación de lípidosExperimental group Lipid peroxidation

Salina + Salina 134.07 ± 5.873Salina + Salina 134.07 ± 5,873

EGb 761 dosis 2.5 mg/kg + Salina 153.85 ± 11.620EGb 761 dose 2.5 mg / kg + Saline 153.85 ± 11.620

EGb 761 dosis 5.0 mg/kg + Salina 151.17 + 5.462EGb 761 dose 5.0 mg / kg + Saline 151.17 + 5,462

EGb 761 dosis 10 mg/kg + Salina 115.93 + 5.682EGb 761 dose 10 mg / kg + Saline 115.93 + 5.682

Salina + MPP+ 231.16 + 9.304 ** EGb 761 dosis 2.5 mg/kg + MPP+ 181.11 + 7.837 +Salina + MPP + 231.16 + 9.304 ** EGb 761 dose 2.5 mg / kg + MPP + 181.11 + 7.837 +

EGb 761 dosis 5.0 mg/kg + MPP+ 143.72 + 12.700 ++EGb 761 dose 5.0 mg / kg + MPP + 143.72 + 12,700 ++

EGb 761 dosis 10 mg/kg + MPP+ 129.84 + 14.920 ++EGb 761 dose 10 mg / kg + MPP + 129.84 + 14.920 ++

Es una gráfica que muestra el efecto benéfico en Ia acción antioxidante y/o antiperoxidante en el cuerpo estriado por Ia administración previa de diferentes dosis de extracto tipo EGb 761 en el parkinsonismo inducido experimentalmente con MPP+. Los resultados son expresados en unidades de fluorescencia/g de tejido húmedo/ml de extracto. Cada grupo representa el promedio + error estándar de n=6-8 experimentos por grupo. ** Estadísticamente diferente del grupo control que es " salina + salina ", p<0.01 , prueba de Tukey. + Estadísticamente diferente del grupo tratado con MPP+ representado por el grupo " salina + MPP+ ", p<0.05, prueba de Tukey. ++ Estadísticamente diferente del grupo tratado con MPP+ representado por el grupo de "salina + MPP+", p<0.01 , prueba de Tukey. EGb 761= extracto de Ginkgo biloba tipo EGb 761; MPP+ = ion 1-metil-4-fenilpiridino. inmediatamente removidos y el cuerpo estriado fue disectado y ensayado para Ia actividad de Ia enzima MAO. La actividad de Ia MAO en el cuerpo estriado se evaluó usando un método previamente descrito (Morinan, A. and Garrat, H. M. 1985. An improved fluorometric assay for brain monoamine oxidase. J. Pharmacol. Methods.13 (3):213-223).It is a graph that shows the beneficial effect on the antioxidant and / or anti-oxidant action in the striatum by the previous administration of different doses of extract type EGb 761 in parkinsonism experimentally induced with MPP + . The results are expressed in fluorescence units / g of wet tissue / ml of extract. Each group represents the average + standard error of n = 6-8 experiments per group. * * Statistically different from the control group that is "saline + saline", p <0.01, Tukey's test. + Statistically different from the group treated with MPP + represented by the group "salina + MPP + ", p <0.05, Tukey's test. ++ Statistically different from the group treated with MPP + represented by the group of "saline + MPP + ", p <0.01, Tukey's test. EGb 761 = Ginkgo biloba extract type EGb 761; MPP + = 1-methyl-4-phenylpyridine ion. immediately removed and the striatum was dissected and tested for the activity of the MAO enzyme. MAO activity in the striatum was evaluated using a previously described method (Morinan, A. and Garrat, HM 1985. An improved fluorometric assay for brain monoamine oxidase. J. Pharmacol. Methods.13 (3): 213-223 ).

El tejido cerebral fue homogenizado en 2.5 mi de agua desionizada a 4°C. Se tomaron alícuotas de 1 mi del homogenado diluido en una relación 1:10 y se les adicionó 0.5 mi de bufer de fosfatos a 0.5 M y un pH de 7.4 conteniendo 100 μg dihidrobromuro de kynuramina como sustrato. Las muestras fueron incubadas a 37°C durante 30 minutos y Ia reacción fue parada por Ia adición de 2 mi de 10% peso/volumen de tricloroacético. Después del enfriamiento y centrifugación a 3,000 x g durante 15 min, una alícuota de 1 mi del sobrenadante se adicionó a 2 mi de una solución de NaOH a una concentración de 1 N. La fluorescencia fue posteriormente medida a 315 nm de excitación y 380 nm de emisión usando un espectrofotómetro para luminiscencia Perkin- Elmer LS 55. En cada experimento, las curvas de calibración fueron construidas, midiendo Ia intensidad de Ia fluorescencia de estándares de Ia 4- hidroxiquinolina que de aquí en adelante denominaremos como 4-HOQ. 4-HOQ es el producto de Ia actividad de Ia MAO en Ia kynuramina. La actividad de Ia MAO fue expresada como μmol de 4-HOQ formada/1 h incubación/g de tejido húmedo.Brain tissue was homogenized in 2.5 ml of deionized water at 4 ° C. Aliquots of 1 ml of the homogenate diluted in a 1:10 ratio were taken and 0.5 ml of phosphate buffer at 0.5 M and a pH of 7.4 were added, containing 100 µg of kynuramine dihydrobromide as substrate. The samples were incubated at 37 ° C for 30 minutes and the reaction was stopped by the addition of 2 ml of 10% w / volume of trichloroacetic. After cooling and centrifugation at 3,000 xg for 15 min, a 1 ml aliquot of the supernatant was added to 2 ml of a NaOH solution at a concentration of 1 N. Fluorescence was subsequently measured at 315nm excitation and 380nm from emission using a Perkin-Elmer LS 55 luminescence spectrophotometer. In each experiment, the calibration curves were constructed, measuring the intensity of the fluorescence of the 4-hydroxyquinoline standards that from now on we will call 4-HOQ. 4-HOQ is the product of the MAO activity in kynuramine. MAO activity was expressed as μmol of 4-HOQ formed / 1 h incubation / g of wet tissue.

Inhibidores específicos de Ia MAO, deprenil para Ia MAO-B y clorgilina para Ia MAO-A, a una concentración de 500 nM en el volumen final y preincubado durante 10 min. Estos fueron usados para medir esas isoformas de Ia MAO solo en el grupo de 6h.Specific MAO inhibitors, deprenil for MAO-B and clorgiline for MAO-A, at a concentration of 500 nM in the final volume and preincubated for 10 min. These were used to measure these MAO isoforms only in the 6h group.

Como se muestra en Ia tabla 2, Ia administración del extracto de Ginkgo biloba tipo EGb 761 a los ratones no produjo incrementos significantes en Ia actividad de Ia MAO total a diferentes tiempos probados de 2, 6, 12 y 24 h, cuando se compara con el grupo tratado con solución salina. En el grupo de ratones que recibieron solo MPP+, Ia actividad total de Ia MAO fue elevada significativamente a 2 y 6 h después de su administración esto es 29% y 30% respectivamente comparado con el grupo control, p<0.05, sin pretratamiento con extracto de Ginkgo biloba tipo EGb 761. Sin embargo, no se observaron cambios en Ia actividad total de Ia MAO a 12 y 24 h después de Ia administración de MPP+ comparado al grupo tratado con solución salina. En contraste, el pretratamiento con extracto de Ginkgo biloba tipo EGb 761 en el grupo con MPP+ previno el incremento de Ia actividad de Ia MAO en el grupo de MPP+ a 6 h, observado sin el pretratamiento. Esto significa que los niveles de Ia actividad de Ia MAO no fueron estadísticamente diferentes de esos de los controles tratados con solución salina. Aunque el pretratamiento con el extracto tipo EGb 761 pareció prevenir el incremento de Ia actividad de Ia MAO producida por el MPP+ a 2 h, pero no fue estadísticamente significativo.As shown in Table 2, the administration of the extract of Ginkgo biloba type EGb 761 to the mice did not produce significant increases in the activity of the total MAO at different times tested of 2, 6, 12 and 24 h, when compared with the group treated with saline. In the group of mice that received only MPP + , the total activity of the MAO was significantly elevated at 2 and 6 h after its administration, that is, 29% and 30% respectively compared to the control group, p <0.05, without pretreatment. with Ginkgo biloba extract type EGb 761. However, no changes in the total activity of the MAO were observed at 12 and 24 h after the administration of MPP + compared to the group treated with saline solution. In contrast, pretreatment with Ginkgo biloba extract type EGb 761 in the MPP + group prevented the increase in MAO activity in the MPP + group at 6 h, observed without pretreatment. This means that the levels of the activity of the MAO were not statistically different from those of the controls treated with saline solution. Although the pretreatment with the EGb 761 type extract seemed to prevent the increase in the activity of the MAO produced by the MPP + at 2 h, but it was not statistically significant.

Como Ia actividad de Ia MAO fue más alta 6 h después de Ia administración de MPP+, nosotros escogimos este punto para el análisis de Ia actividad de Ia MAO-A y MAO-B. Como se observa en Ia figura 1 , el extracto tipo EGb 761 no tuvo efectos estadísticamente significativos en Ia actividad de Ia MAO-B comparado al grupo tratado con salina. El MPP+ incremento Ia actividad de Ia MAO-B por 70% sobre el grupo de salina tratado con MPP+, p<0.05. El pretratamiento con extracto tipo EGb 761 produjo significante protección contra Ia neurotoxicidad con MPP+, por reducir parcialmente el incremento en Ia actividad de Ia MAO-B a casi los niveles normales cuando se compara al grupo tratado con solución salina. En Ia figura 1 se puede observar que el análisis de Ia MAO-A en grupos adicionales no mostró cambios en su actividad para todos los grupos tratados.As the activity of the MAO was higher 6 h after the administration of MPP + , we chose this point for the analysis of the activity of the MAO-A and MAO-B. As observed in Figure 1, the EGb 761 type extract had no statistically significant effects on the activity of MAO-B compared to the group treated with saline. The MPP + increased the MAO-B activity by 70% over the saline group treated with MPP + , p <0.05. Pretreatment with extract type EGb 761 produced significant protection against neurotoxicity with MPP + , by partially reducing the increase in MAO-B activity to almost normal levels when compared to the group treated with saline solution. In Figure 1 it can be seen that the analysis of MAO-A in additional groups did not show changes in its activity for all the treated groups.

Ejemplo 5Example 5

Esta invención muestra claramente que el tratamiento con un extracto de Ginkgo biloba tipo EGb 761 incrementa Ia actividad de la tirosina hidroxilasa. enzima determinante en Ia síntesis de dopamina como respuesta protectora en el parkinsonismo inducido con MPP+. Se diseñaron 4 grupos experimentales y fueron tratados experimentalmente exactamente como se describió en el ejemplo 3. Posteriormente analizé Ia actividad de Ia tirosina hidroxilasa. Después de Ia manipulación experimental los ratones fueron sacrificados por dislocación cervical a 2, 6, 12 y 24 h después de Ia administración de MPP+. TABLA 2. Actividad de Ia MAO total en el cuerpo estriadoThis invention clearly shows that treatment with an extract of Ginkgo biloba type EGb 761 increases the activity of tyrosine hydroxylase. determining enzyme in the synthesis of dopamine as a protective response in parkinsonism induced with MPP + . 4 experimental groups were designed and were experimentally treated exactly as described in Example 3. Later, I analyzed the activity of the tyrosine hydroxylase. After the experimental manipulation, the mice were sacrificed by cervical dislocation at 2, 6, 12 and 24 h after the administration of MPP + . TABLE 2. Activity of the total MAO in the striatum

Grupo experimental Actividad de Ia MAO totalExperimental group Activity of the total MAO

2 horas2 hours

Salina + Salina 3.602 ± 0.202 EGb761 + Salina 3.635 + 0.333 Salina + MPP+ 4.627 ± 0.112 * EGb761 + MPP+ 4.001 ± 0.325Salina + Salina 3,602 ± 0.202 EGb761 + Salina 3,635 + 0.333 Salina + MPP + 4,627 ± 0.112 * EGb761 + MPP + 4,001 ± 0.325

6 horas6 hours

Salina + Salina 4.247 ± 0.147 EGb761+Salina 4.339 ± 0.097 Salina + MPP+ 5.490 ± 0.418 * EGb761 + MPP+ 4.179 ± 0.406 +Salina + Salina 4,247 ± 0.147 EGb761 + Salina 4,339 ± 0.097 Salina + MPP + 5,490 ± 0.418 * EGb761 + MPP + 4,179 ± 0.406 +

12 horas12 hours

Salina+Salina 3.627 ± 0.371 EGb761+Salina 3.775 ± 0.246 Salina + MPP+ 3.467 ± 0.214 EGb761 + MPP+ 3.415 ± 0.487Salina + Salina 3,627 ± 0.371 EGb761 + Salina 3,775 ± 0.246 Salina + MPP + 3,467 ± 0.214 EGb761 + MPP + 3,415 ± 0.487

24 horas24 hours

Salina + Salina 3.972 ± 0.155 EGb761 + Salina 4.197 ± 0.190 Salina + MPP+ 4.171 ± 0.187 EGb761 + MPP+ 4.052 + 0.165Salina + Salina 3,972 ± 0.155 EGb761 + Salina 4,197 ± 0.190 Salina + MPP + 4,171 ± 0.187 EGb761 + MPP + 4,052 + 0.165

Esta tabla muestra los efectos benéficos de Ia regulación y/o inhibición de Ia actividad enzimática de Ia MAO total en el cuerpo estriado por pretratamiento con extracto de Ginkgo biloba tipo EGb 761 en el parkinsonismo inducido experimentalmente con MPP+. Los resultados son expresados en actividad de Ia MAO total en μmoles 4-OHQ/g/h. Cada grupo representa el promedio +/- error estándar de Ia media de 6-8 experimentos por grupo. * Estadísticamente diferente del grupo control representado por el grupo de "salina + salina", prueba de Tukey. +Estadísticamente diferente del grupo tratado con MPP+ representado por el grupo de "salina + MPP+", p<0.05, prueba de Tukey. Los cerebros de los ratones fueron inmediatamente removidos y los cuerpos estriados se disectaron. Una alícuota de 500 μl de solución de perclórico-metabisulfito de sodio a 0.1% peso/volumen fue adicionada al peso del tejido y sonicada con un sistema labsonic modelo Lab-line instruments, Melrose Park.IL. Posteriormente las muestras fueron centrifugadas a 4, 000 g durante 10 min y los sobrenadantes fueron mantenidos a -70 0C hasta su análisis.This table shows the beneficial effects of the regulation and / or inhibition of the enzymatic activity of the total MAO in the striatum by pretreatment with Ginkgo biloba extract type EGb 761 in parkinsonism experimentally induced with MPP + . The results are expressed in activity of the total MAO in μmoles 4-OHQ / g / h. Each group represents the average +/- standard error of the average of 6-8 experiments per group. * Statistically different from the control group represented by the "saline + saline" group, Tukey's test. + Statistically different from the group treated with MPP + represented by the group of "saline + MPP + ", p <0.05, Tukey's test. The brains of the mice were immediately removed and the striatum was dissected. A 500 µl aliquot of 0.1% w / v sodium perchloric-metabisulfite solution was added to the tissue weight and sonicated with a Labsonic Lab-line instruments model system, Melrose Park.IL. Subsequently the samples were centrifuged at 4,000 g for 10 min and the supernatants were kept at -70 0 C until analysis.

La actividad de Ia tirosina hidroxilasa fue ensayada en el cuerpo estriado usando un sistema de cromatografía de líquidos de alta resolución modelo series 200 LC de Perkin-Elmer, conectado a un detector ESA Coulochem 5100A con los siguientes potenciales del electrodo de referencia de +0.4 V y oxidación de Ia CeWa1ZCeWa2 +0.03/+0.35 V. (Larsson L-G, Rényi L, Ross SB, Svensson B, and Λngeby-Moller K.1990. Different effects on the responses of functional pre- and postsynaptic 5-HTIA receptors by repeated treatment of rats with the 5-HTIA receptor agonist 8-OH-DPAT. Neuropharmacology. 29: 85-91). Para medir esta actividad medí Ia acumulación de Ia L-DOPA 30 minutos después de Ia administración de un inhibidor de Ia L-DOPA descarboxilasa que actúa centralmente. El inhibidor usado fue el NSD 1015 a una dosis de 100 mg/kg (Kehr, W., Carlsson, A., Lindquist, M., Magnusson, T., and Atack, C. 1972. Evidence for a receptor mediated feedback control of striatal tyrosine hydroxylase activity. J. Pharm. Pharmacol. 24: 744-747).The activity of the tyrosine hydroxylase was assayed in the striatum using a Perkin-Elmer model series 200 LC high-performance liquid chromatography system, connected to an ESA Coulochem 5100A detector with the following reference electrode potentials of +0.4 V and oxidation of Ia CeWa 1 ZCeWa 2 + 0.03 / + 0.35 V. (Larsson LG, Rényi L, Ross SB, Svensson B, and Λngeby-Moller K. 1990. Different effects on the responses of functional pre- and postsynaptic 5-HTIA receptors by repeated treatment of rats with the 5-HTIA receptor agonist 8-OH-DPAT. Neuropharmacology. 29: 85-91). To measure this activity, I measured the accumulation of L-DOPA 30 minutes after the administration of a centrally acting L-DOPA decarboxylase inhibitor. The inhibitor used was NSD 1015 at a dose of 100 mg / kg (Kehr, W., Carlsson, A., Lindquist, M., Magnusson, T., and Atack, C. 1972. Evidence for a receptor mediated feedback control of striatal tyrosine hydroxylase activity. J. Pharm. Pharmacol. 24: 744-747).

Las curvas de calibración fueron construidas para Ia dopamina, L-DOPA y Ia concentración fue obtenida por interpolación de Ia respectiva curva estándar. Se empleó una columna analítica de catecolaminas de 100 x 4.8 mm con un tamaño de partícula de 3 μm marca Alltech Associates, Inc. La fase móvil consisitió de un bufer de fosfatos acuoso a un pH de 3.1 , el cual contenía octil sufato de sodio a 0.2 mM, EDTA a 0.1 mM, metanol 15% volumen/volumen, a un pH de 2.6. Como se muestra en Ia tabla 3, el extracto tipo EGb 761 sólo no influye en Ia actividad de Ia tirosina hidroxilasa a todos los tiempos probados cuando sólo se compara con el grupo de " salina + salina ". La administración de MPP+ a 12 y 24 h mostró una pérdida de actividad de Ia tirosina hidroxilasa en el estriado de 34% y 33% respectivamente, versus el grupo de "salina + salina", pero este extracto bloqueó esta pérdida en el grupo de " EGb 761 + MPP+ ".The calibration curves were built for dopamine, L-DOPA and the concentration was obtained by interpolation of the respective standard curve. An analytical column of catecholamines of 100 x 4.8 mm with a particle size of 3 μm, used by Alltech Associates, Inc. was used. The mobile phase consisted of an aqueous phosphate buffer at a pH of 3.1, which contained sodium octyl sufate at 0.2 mM, EDTA at 0.1 mM, methanol 15% volume / volume, at a pH of 2.6. As shown in Table 3, the EGb 761 type extract only does not influence the activity of the tyrosine hydroxylase at all the times tested when it is only compared with the "saline + saline" group. The administration of MPP + at 12 and 24 h showed a loss of activity of the tyrosine hydroxylase in the Striated 34% and 33% respectively, versus the "saline + saline" group, but this extract blocked this loss in the "EGb 761 + MPP + " group.

Ejemplo 6Example 6

En esta invención se observa que el tratamiento con extracto de Ginkαo biloba tipo EGb 761 incrementa el contenido de dopamina en el cuerpo estriado como respuesta protectora en el parkinsonismo inducido con MPP+. Analizé el contenido de dopamina sólo en los grupos de ratones tratados con EGb 761 a una dosis de 10 mg/kg, vía i. p. durante 17 días y a una dosis de MPP+ de 0.72 mg/kg, con los respectivos grupo control. Los ratones fueron sacrificados por dislocación cervical a 2, 6, 12 y 24 h después de Ia administración de MPP+ como se mencionó en el ejemplo 3. Los cerebros de los ratones fueron inmediatamente removidos y los cuerpos estriados se disectaron. Las concentraciones de Ia dopamina fueron analizadas de acuerdo al método previamente descrito (Larsson L-G, Rényi L, Ross SB, Svensson B, and Ángeby-Móller K.1990. Different effects on the responses of functional pre- and postsynaptic 5-HT1A receptors by repeated treatment of rats with the 5-HT-IA receptor agonist 8-OH-DPAT. Neuropharmacology. 29: 85-91). Una alícuota de 500 μl de solución de perclórico-metabisulfito de sodio a 0.1% peso/volumen fue adicionada al peso del tejido y sonicada con un sistema labsonic marca Lab-line Instruments, Melrose Park.IL. Posteriormente las muestras fueron centrifugadas a 4, 000 g durante 10 min y los sobrenadantes fueron mantenidos a -70 0C hasta su análisis. El contenido de Ia dopamina en el cuerpo estriado fue analizado usando un sistema de cromatografía de alta resolución con modelo series LC 200 de Perkin-Elmer, conectado a un detector ESA Coulochem 5100A, como se describió en el ejemplo 5. El contenido de dopamina en el cuerpo estriado después de Ia administración de MPP+ se muestra en tabla 4. La administración de extracto de Ginkgo biloba tipo EGb 761 a los ratones no produjo alteración significativa en el contenido de dopamina a todos los tiempos probados cuando son comparados a los animales control representados por el grupo " salina + salina ". Los ratones en el grupo de " salina + MPP+ " a 12 y 24 h presentaron TABLA 3. Actividad de Ia tirosina hidroxilasa en cuerpo estriadoIn this invention, it is observed that the treatment with extract of Ginkαo biloba type EGb 761 increases the dopamine content in the striatum as a protective response in parkinsonism induced with MPP + . I analyzed the dopamine content only in the groups of mice treated with EGb 761 at a dose of 10 mg / kg, ip for 17 days and at an MPP + dose of 0.72 mg / kg, with the respective control group. The mice were sacrificed by cervical dislocation at 2, 6, 12 and 24 h after the administration of MPP + as mentioned in Example 3. The brains of the mice were immediately removed and the striatum was dissected. Dopamine concentrations were analyzed according to the previously described method (Larsson LG, Rényi L, Ross SB, Svensson B, and Ángeby-Móller K. 1990. Different effects on the responses of functional pre- and postsynaptic 5-HT 1A receptors by repeated treatment of rats with the 5-HT-IA receptor agonist 8-OH-DPAT. Neuropharmacology. 29: 85-91). A 500 µl aliquot of 0.1% w / v sodium perchloric-metabisulfite solution was added to the tissue weight and sonicated with a Labson line brand Labsonic system, Melrose Park.IL. Subsequently the samples were centrifuged at 4,000 g for 10 min and the supernatants were kept at -70 0 C until analysis. The content of dopamine in the striatum was analyzed using a Perkin-Elmer LC 200 series model high-resolution chromatography system, connected to an ESA Coulochem 5100A detector, as described in Example 5. The content of dopamine in the striatum after the administration of MPP + is shown in table 4. The administration of Ginkgo biloba extract type EGb 761 to the mice did not produce a significant alteration in the content of dopamine at all the times tested when compared to the control animals. represented by the group "salina + salina". Mice in the "saline + MPP + " group at 12 and 24 h had TABLE 3. Activity of Tyrosine Hydroxylase in Striatum

Grupo experimental Actividad de Ia tirosina hidroxilasaExperimental group Activity of tyrosine hydroxylase

2 horas2 hours

Salina+Salina 0.787 ± 0.043 EGb761 + Salina 0.864 ± 0.027 Salina + MPP+ 0.882 ± 0.073 EGb761 + MPP+ 0.924 ± 0.071Salina + Salina 0.787 ± 0.043 EGb761 + Salina 0.864 ± 0.027 Salina + MPP + 0.882 ± 0.073 EGb761 + MPP + 0.924 ± 0.071

6 horas6 hours

Salina + Salina 0.731 ± 0.050 EGb761+Salina 0.739 ± 0.131 Salina + MPP+ 0.635 ± 0.076 EGb761 + MPP+ 0.642 ± 0.020Salina + Salina 0.731 ± 0.050 EGb761 + Salina 0.739 ± 0.131 Salina + MPP + 0.635 ± 0.076 EGb761 + MPP + 0.642 ± 0.020

12 horas12 hours

Salina+Salina 0.658 ± 0.041 EGb761+Salina 0.650 ± 0.057 Salina + MPP+ 0.432 ± 0.065 * EGb761 + MPP+ 0.717 ± 0.064 +Salina + Salina 0.658 ± 0.041 EGb761 + Salina 0.650 ± 0.057 Salina + MPP + 0.432 ± 0.065 * EGb761 + MPP + 0.717 ± 0.064 +

24 horas24 hours

Salina + Salina 0.756 ± 0.065 EGb761 + Salina 0.729 ± 0.042 Salina + MPP+ 0.509 ± 0.051 * EGb761 + MPP+ 0.951 ± 0.123 +Salina + Salina 0.756 ± 0.065 EGb761 + Salina 0.729 ± 0.042 Salina + MPP + 0.509 ± 0.051 * EGb761 + MPP + 0.951 ± 0.123 +

Esta tabla muestra el incremento de Ia actividad de Ia tirosina hidroxilasa en el cuerpo estriado por el pretratamiento con extracto tipo EGb 761 como respuesta protectora en el parkinsonismo inducido experimentalmente con MPP+. Los resultados son expresados como actividad de Ia tirosina hidroxilasa en μg/g de tejido. Cada grupo representa el promedio +/- error estándar de Ia media de 6-8 experimentos por grupo. * Estadísticamente diferente del grupo control representado como " salina + salina ", p<0.05, prueba de Tukey. + Estadísticamente diferente del grupo tratado con MPP+ representado como " salina + MPP+", p<0.05, prueba de Tukey. una reducción de Ia dopamina de 30% y 36%, respectivamente como resultado de Ia acción tóxica del MPP+. No se encontraron alteraciones a 2 h después de Ia administración de MPP+. Pero una tendencia a Ia reducción es clara en el grupo de MPP+ a las 6 h después de su administración pero no estadísticamente significativa.This table shows the increase in tyrosine hydroxylase activity in the striatum by pretreatment with EGb 761-type extract as a protective response in parkinsonism experimentally induced with MPP + . The results are expressed as tyrosine hydroxylase activity in µg / g of tissue. Each group represents the average +/- standard error of the average of 6-8 experiments per group. * Statistically different from the control group represented as "saline + saline", p <0.05, Tukey's test. + Statistically different from the group treated with MPP + represented as "saline + MPP + ", p <0.05, Tukey's test. a reduction of dopamine of 30% and 36%, respectively, as a result of the toxic action of MPP + . No alterations were found at 2 h after the administration of MPP + . But a trend towards reduction is clear in the MPP + group at 6 h after administration, but not statistically significant.

Esos decrementos significantes en Ia concentración de dopamina a 12 h y 24 h pero no a 2 h y 6 h, sugiere que el incremento de Ia actividad de Ia MAO precede al efecto disminuidor de Ia dopamina producida por el MPP+. El p retratamiento con extracto tipo EGb 761 seguido por Ia administración del MPP+ parcialmente previno el efecto disminuidor de Ia dopamina producido por el MPP+ a 12 h en 31% y a 24 h con 32% de protección comparado al grupo del MPP+.These significant decreases in the concentration of dopamine at 12 h and 24 h but not at 2 h and 6 h, suggests that the increase in the activity of MAO precedes the decreasing effect of dopamine produced by MPP + . The retreatment with extract type EGb 761 followed by the administration of MPP + partially prevented the diminishing effect of dopamine produced by MPP + at 12 h in 31% and at 24 h with 32% protection compared to the MPP + group .

Ejemplo 7Example 7

En esta invención encontré que el extacto de Ginkαo biloba tipo EGb 761 restaura a las neuronas dopaminérgicas del cuerpo estriado después de que se induce el parkinsonismo con MPTP. Se diseñaron 4 grupos experimentales: grupo I: solución salina por vía i.p + solución salina por vía i.p; grupo II: solución salina por vía i.p + EGb 761 por vía i.p; grupo III: MPTP por vía ip + solución salina por vía i.p.; grupo IV: MPTP por vía i.p. + EGb 761 por vía i.p.In this invention I found that the Ginkαo biloba type EGb 761 extract restores striatal dopaminergic neurons after parkinsonism is induced with MPTP. Four experimental groups were designed: group I: saline solution via i.p + saline solution via i.p route; group II: saline by the i.p + EGb 761 route by the i.p route; group III: MPTP via ip + saline via i.p .; Group IV: MPTP via i.p. + EGb 761 via i.p.

Los animales de los grupos I y Il recibieron solución salina por vía i.p y los grupos III y IV recibieron MPTP a una dosis de 30 mg/kg, por vía i.p. diariamente durante 5 días. Después del parkinsonismo inducido por Ia MPTP los animales de los grupos Il y IV fueron administrados con extracto tipo EGb 761 a una dosis de 40 ó 120 mg/kg durante 18 días ó se administró solución salina en los grupos I y III utilizándolos como grupo control. Posteriormente los animales fueron sacrificados por dislocación cervical después de Ia última administración de extracto tipo. EGb 761. Los cerebros fueron inmediatamente removidos y el cuerpo estriado fue disectado y se analizó el contenido de dopamina en el cuerpo estriado por cromatografía de líquidos de alta resolución como se describió en el ejemplo 6. TABLA 4. Contenido de dopamina en cuerpo estriadoAnimals in groups I and Il received saline ip and groups III and IV received MPTP at a dose of 30 mg / kg, ip daily for 5 days. After MPTP-induced parkinsonism, animals in groups Il and IV were administered with EGb 761 type extract at a dose of 40 or 120 mg / kg for 18 days or saline was administered in groups I and III using them as a control group. . Subsequently, the animals were sacrificed by cervical dislocation after the last administration of the type extract. EGb 761. The brains were immediately removed and the striatum was dissected, and the dopamine content in the striatum was analyzed by high-performance liquid chromatography as described in Example 6. TABLE 4. Dopamine content in striatum

Grupo experimental Contenido de dopaminaExperimental group Dopamine content

2 horas2 hours

Salina+Salina 10.77 ± 0.436Salina + Salina 10.77 ± 0.436

EGb761 + Salina 11.00 ± 0.666EGb761 + Salina 11.00 ± 0.666

Salina + MPP+ 11.97 ± 0.389 EGb761 + MPP+ 10.60 ± 0.468Salina + MPP + 11.97 ± 0.389 EGb761 + MPP + 10.60 ± 0.468

6 horas6 hours

Salina + Salina 9.537 ± 0.802Salina + Salina 9,537 ± 0.802

EGb761+Salina 9.655 ± 1.035EGb761 + Salina 9,655 ± 1,035

Salina + MPP+ 7.640 ± 0.254 EGb761 + MPP+ 10.40 ± 1.208Salina + MPP + 7,640 ± 0.254 EGb761 + MPP + 10.40 ± 1,208

12 horas12 hours

Salina+Salina 9.732 ± 0.543Salina + Salina 9,732 ± 0.543

EGb761+Salina 9.961 ± 1.070EGb761 + Salina 9,961 ± 1,070

Salina + MPP+ 6.778 ± 0.497 * EGb761 + MPP+ 8.885 ± 0.675 +Salina + MPP + 6,778 ± 0.497 * EGb761 + MPP + 8,885 ± 0.675 +

24 horas24 hours

Salina + Salina 10.44 ± 0.330Salina + Salina 10.44 ± 0.330

EGb761 + Salina 11.05 ± 0.156EGb761 + Salina 11.05 ± 0.156

Salina + MPP+ 6.68 ± 0.429 ** EGb761 + MPP+ 8.81 ± 0.639 +Salina + MPP + 6.68 ± 0.429 ** EGb761 + MPP + 8.81 ± 0.639 +

Esta tabla muestra Ia protección que produce el pretratamiento con extracto tipo EGb 761 en los niveles de dopamina en el cuerpo estriado en el parkinsonismo inducido experimentalmente con MPP+. Los resultados son expresados en contenido de dopamina en μg/g de tejido. Cada grupo representa el promedio de +/- error estándar de Ia media de 6-8 experimentos por grupo. * Estadísticamente diferente del grupo control representado como " salina + salina ", p<0.05, prueba de Tukey. ** Estadísticamente diferente de control representado como "salina + salina", p<0.01 , prueba de Tukey. + Estadísticamente diferente del grupo tratado con MPP+ representado como " salina + MPP+ ", p<0.05, prueba de Tukey. MPP+=I -metil-4-fenilpiridino. El contenido de dopamina en el cuerpo estriado después de Ia administración de extracto tipo EGb 761 en animales con parkinsonismo inducido con MPTP se muestran en Ia figura 2. Como se muestra en Ia figura 2 Ia administración del extracto tipo EGb 761 a dosis de 40 ó 120 mg/kg a los ratones, no produjo alteración significante en el contenido de dopamina cuando son comparados a los animales control denominado como "salina + salina". Los ratones en el grupo de "salina + MPTP" presentaron una reducción de Ia dopamina de 48% como resultado de Ia acción tóxica de Ia MPTP, ver Ia figura 2. El efecto neurorestaurador del extracto tipo EGb 761 en animales con parkinsonismo inducido por MPTP previno parcialmente el efecto disminuidor de Ia dopamina producido por Ia MPTP, 70% y 40% a una dosis del extracto tipo EGb 761 de 40 y 120 mg/kg respectivamente.This table shows the protection produced by pretreatment with extract type EGb 761 in the levels of dopamine in the striatum in parkinsonism experimentally induced with MPP + . The results are expressed in dopamine content in µg / g of tissue. Each group represents the average of +/- standard error of the average of 6-8 experiments per group. * Statistically different from the control group represented as "saline + saline", p <0.05, Tukey's test. ** Statistically different from control represented as "saline + saline", p <0.01, Tukey test. + Statistically different from the group treated with MPP + represented as "saline + MPP + ", p <0.05, Tukey's test. MPP + = I -methyl-4-phenylpyridine. The dopamine content in the striatum after the administration of extract type EGb 761 in animals with parkinsonism induced with MPTP is shown in Figure 2. As shown in Figure 2, the administration of extract type EGb 761 at doses of 40 or 120 mg / kg to the mice, did not produce significant alteration in the dopamine content when compared to the control animals named as "saline + saline". Mice in the "saline + MPTP" group presented a 48% reduction in dopamine as a result of the toxic action of MPTP, see Figure 2. The neuro-restorative effect of EGb 761-type extract in animals with parkinsonism induced by MPTP It partially prevented the decreasing effect of dopamine produced by MPTP, 70% and 40% at a dose of EGb 761 type extract of 40 and 120 mg / kg respectively.

Por Io dicho anteriormente, se puede afirmar que estas características del extracto de Ginkgo biloba descrito no han sido logradas por ningún otro fármaco y reúne en si las características de prevenir y de revertir el daño celular producido durante Ia enfermedad de Parkinson. Based on what was said previously, it can be affirmed that these characteristics of the Ginkgo biloba extract described have not been achieved by any other drug and it brings together the characteristics of preventing and reversing the cellular damage produced during Parkinson's disease.

Claims

ReivindicacionesHabiendo descrito suficientemente mi invención considero como una novedad y por Io tanto reclamo como de mi exclusiva propiedad, Io contenido en las siguientes cláusulas: Claims Having sufficiently described my invention, I consider it a novelty and therefore I claim as well as my exclusive property, contained in the following clauses: 1. El uso de una composición farmacéutica para el tratamiento de Ia enfermedad de Parkinson que es obtenido de un extracto de Ginkgo hiloba que comprende 20 a 30% de glicósidos de flavonoides, 2.5 a 4.5% de ginkgólidos A, B, C y J, de 2.0 a 4.0% de bilobálidos, menos de 10% de proantocianidinas y menos de 10 ppm de compuestos de tipo alquifenol, todos los porcentajes son por peso.1. The use of a pharmaceutical composition for the treatment of Parkinson's disease that is obtained from an extract of Ginkgo filaba which comprises 20 to 30% of flavonoid glycosides, 2.5 to 4.5% of ginkgolides A, B, C and J, from 2.0 to 4.0% of bilobalides, less than 10% of proanthocyanidins and less than 10 ppm of compounds of the alkylphenol type, all the percentages are by weight. 2. El extracto de Ia reivindicación 1 contiene de 22 a 26% de glicósidos de flavonoides de porcentaje en peso.2. The extract of claim 1 contains from 22 to 26% of flavonoid glycosides of percentage by weight. 3. El extracto de Ia reivindicación 1 contiene menos de 1 ppm de compuestos alquifenoles.3. The extract of claim 1 contains less than 1 ppm of alkyphenol compounds. 4. Un extracto de las hojas de Ginkgo biloba el cual es sustancialmente libre de compuestos alquifenoles y que tiene un alto contenido de glicósidos de flavonoides y comprende sustancialmente todo tipo de ginkgólidos y bilobálidos originalmente presentes en las hojas.4. An extract from the leaves of Ginkgo biloba which is substantially free of alkyphenol compounds and which is high in flavonoid glycosides and comprises substantially all kinds of ginkgolides and bilobalides originally present in the leaves. 5. Una composición farmacéutica de un extracto de las hojas Ginkgo biloba conteniendo 20 a 30%% de glicósidos de flavonoides, 2.5 a 4.5% de ginkgólidos seleccionados del ginkgólido A, B, C y J o mezcla de ellos, 2.0 a 4.0% de bilobálidos, menos de 10 ppm de compuestos alquifenoles y menos de 10% de proantocianidinas.5. A pharmaceutical composition of an extract of the Ginkgo biloba leaves containing 20 to 30 %% of flavonoid glycosides, 2.5 to 4.5% of ginkgolides selected from ginkgolide A, B, C and J, 2.0 to 4.0% of bilobalides, less than 10 ppm of alkyphenols and less than 10% of proanthocyanidins. 6. Una composición farmacéutica de Ia reivindicación 5 donde el extracto contiene de 22 a 26% de glicósidos de flavonoides. 6. A pharmaceutical composition of claim 5 wherein the extract contains from 22 to 26% of flavonoid glycosides. 7. Una composición farmacéutica de Ia reivindicación 5 donde el extracto preparado contiene menos de 1 ppm de compuestos alquifenoles.7. A pharmaceutical composition of claim 5 wherein the prepared extract contains less than 1 ppm of alkyphenol compounds. 8. Una composición farmacéutica para el tratamiento de Ia enfermedad de Parkinson que comprende un extracto de Ginkgo biloba de Ia reivindicación 1 , 2, 3, 5, 6 y 7 que contiene un acarreador apropiado donde Ia concentración del extracto es suficiente para aliviar Ia enfermedad de Parkinson.8. A pharmaceutical composition for the treatment of Parkinson's disease comprising an extract of Ginkgo biloba of claim 1, 2, 3, 5, 6 and 7 that contains an appropriate carrier where the concentration of the extract is sufficient to alleviate the disease. Parkinson's disease. 9. Una composición farmacéutica de Ia reivindicación 8 donde el extracto de Ginkgo hiloba es administrado en una cantidad de 0.1 mg a 1 g por día dependiendo de Ia gravedad de Ia enfermedad.9. A pharmaceutical composition of claim 8 where the Ginkgo filaba extract is administered in an amount of 0.1 mg to 1 g per day depending on the severity of the disease. 10. Una composición farmacéutica de Ia reivindicación 8 donde el extracto de Ginkgo biloba es administrado en una cantidad de 5 mg a 600 mg por día.10. A pharmaceutical composition of claim 8 where the Ginkgo biloba extract is administered in an amount of 5 mg to 600 mg per day. 11. Una composición farmacéutica de Ia reivindicación 8 donde el extracto de Ginkgo biloba es administrado en una cantidad de 10 mg a 100 mg por día.11. A pharmaceutical composition of claim 8 where the Ginkgo biloba extract is administered in an amount of 10 mg to 100 mg per day. 12. Una composición farmacéutica de Ia reivindicación 8 donde el extracto de Ginkgo biloba es administrado en una cantidad de 10 mg a 300 mg por día.12. A pharmaceutical composition of claim 8 where the Ginkgo biloba extract is administered in an amount of 10 mg to 300 mg per day. 13. Una composición farmacéutica de Ia reivindicación 8 donde Ia donde Ia preparación farmacéutica es administrada por vía tópica, oral, ruta parenteral, por inyección como intramuscular, subcutánea, intravenosa, etc.13. A pharmaceutical composition of claim 8 where the pharmaceutical preparation is administered topically, orally, parenterally, by injection such as intramuscular, subcutaneous, intravenous, etc. 14. Una composición farmacéutica de Ia reivindicación 8 donde Ia donde Ia preparación farmacéutica puede estar en forma de sólidos, por ejemplo polvos, tabletas, cápsulas, liposomas o supositorios.14. A pharmaceutical composition of claim 8 wherein the pharmaceutical preparation may be in the form of solids, for example powders, tablets, capsules, liposomes or suppositories. 15. Una composición farmacéutica de Ia reivindicación 8 donde Ia preparación farmacéutica puede estar en forma de un líquido, por ejemplo, soluciones, emulsiones, suspensiones o jarabe. 15. A pharmaceutical composition of claim 8 where the pharmaceutical preparation may be in the form of a liquid, for example, solutions, emulsions, suspensions or syrup. 16. Una composición farmacéutica de Ia reivindicación 8 que es capaz de incrementar Ia actividad de Ia tirosina hidroxilasa en el cerebro.16. A pharmaceutical composition of claim 8 that is capable of increasing the activity of tyrosine hydroxylase in the brain. 17. Una composición farmacéutica de Ia reivindicación 8 que es capaz de actuar como agente antiperoxidante y/o antioxidante en el cerebro.17. A pharmaceutical composition of claim 8 that is capable of acting as an anti-oxidant and / or antioxidant agent in the brain. 18. Una composición farmacéutica de Ia reivindicación 8 que incrementa los niveles de dopamina en el cerebro.18. A pharmaceutical composition of claim 8 that increases the levels of dopamine in the brain. 19. Una composición farmacéutica de Ia reivindicación 8 que actúa como un inhibidor y/o regulador de Ia enzima monoamino oxidasa tipo A y B.19. A pharmaceutical composition of claim 8 that acts as an inhibitor and / or regulator of the monoamine oxidase enzyme type A and B. 20. Una composición farmacéutica de Ia reivindicación 8 que actúa como un inhibidor y/o regulador de Ia enzima monoamino oxidasa total. 20. A pharmaceutical composition of claim 8 that acts as an inhibitor and / or regulator of the total monoamine oxidase enzyme.
PCT/MX2004/000042 2004-07-02 2004-07-02 Use of ginkgo biloba extract for the preparation of a medicine that is used to treat parkinson's disease Ceased WO2006004386A1 (en)

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Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0431536A1 (en) * 1989-12-04 1991-06-12 Dr. Willmar Schwabe GmbH &amp; Co. A method of preparation of an extract from Gingko biloba leaves.
EP0431535A1 (en) * 1989-12-04 1991-06-12 Dr. Willmar Schwabe GmbH &amp; Co. Extract from Ginkgo biloba leaves, its method of preparation and pharmaceuticals containing the extract
WO2000007592A1 (en) * 1998-08-07 2000-02-17 Societe De Conseils De Recherches Et D'applications Scientifiques (S.C.R.A.S.) Use of ginkgo biloba extracts for preparing a medicine for treating amyotrophic lateral sclerosis
WO2000064462A1 (en) * 1999-04-21 2000-11-02 Yuyu Industrial Co., Ltd. Pharmaceutical compositions containing selegiline and ginkgo biloba extract useful for dementia
WO2002083158A1 (en) * 2001-04-10 2002-10-24 Societe De Conseils De Recherches Et D'applications Scientifiques (S.C.R.A.S.) Method for preparing an extract of ginkgo biloba leaves highly enriched in active principles
WO2004014405A1 (en) * 2002-07-16 2004-02-19 Societe De Conseils De Recherches Et D'applications Scientifiques (S.C.R.A.S.) Use of ginkgo biloba extracts in order to promote muscle mass to the detriment of fatty mass

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0431536A1 (en) * 1989-12-04 1991-06-12 Dr. Willmar Schwabe GmbH &amp; Co. A method of preparation of an extract from Gingko biloba leaves.
EP0431535A1 (en) * 1989-12-04 1991-06-12 Dr. Willmar Schwabe GmbH &amp; Co. Extract from Ginkgo biloba leaves, its method of preparation and pharmaceuticals containing the extract
WO2000007592A1 (en) * 1998-08-07 2000-02-17 Societe De Conseils De Recherches Et D'applications Scientifiques (S.C.R.A.S.) Use of ginkgo biloba extracts for preparing a medicine for treating amyotrophic lateral sclerosis
WO2000064462A1 (en) * 1999-04-21 2000-11-02 Yuyu Industrial Co., Ltd. Pharmaceutical compositions containing selegiline and ginkgo biloba extract useful for dementia
WO2002083158A1 (en) * 2001-04-10 2002-10-24 Societe De Conseils De Recherches Et D'applications Scientifiques (S.C.R.A.S.) Method for preparing an extract of ginkgo biloba leaves highly enriched in active principles
WO2004014405A1 (en) * 2002-07-16 2004-02-19 Societe De Conseils De Recherches Et D'applications Scientifiques (S.C.R.A.S.) Use of ginkgo biloba extracts in order to promote muscle mass to the detriment of fatty mass

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
CAO F ET AL: "[Pattern recognition applied to the fingerprint of traditional Chinese medicine characterized by two-dimension information data]", JOURNAL OF HUAZHONG UNIVERSITY OF SCIENCE AND TECHNOLOGY MEDICAL SCIENCES, vol. 23, no. 2, 2003, pages 151 - 153 *
HEMMER R ET AL: "Zur cerebralen wirksamkeit eines pflanzenpraoarates aus ginkgo biloba", ARZNEIMITTEL FORSHUNG, vol. 17, no. 4, 1967, pages 491 - 494 *
ROJAS P ET AL: "EGb761 blocks MPP+-induced lipid peroxidation in mouse corpus striatum", NEUROCHEMICAL RESEARCH, vol. 26, no. 11, 2001, pages 1245 - 1251 *
WU WEU-RAN ET AL: "Involvement of monoamine oxidase inhibition in neuroprotective and neurorestorative effects of Ginkgo biloba extract against MPTP-induced nigrostriatal dopaminergic toxicity in C57 mice.", LIFE SCIENCES, vol. 65, no. 2, 1999, pages 157 - 164 *
YANG SU FEN ET AL: "Protective effect and mechanism of ginkgo biloba leaf extracts for parkinson didease induced by 1-methyl-4phenyl-1,2,3,6-tetrahydrpyridine", ACTA PHARMACOLOGICA CINICA, vol. 22, no. 12, 2001, pages 1089 - 1093 *

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