WO2009064151A1

WO2009064151A1 - Method of obtaining a dry extract of galphimia glauca, pharmaceutical compositions comprising said extract and use of the compositions to treat anxiety

Info

Publication number: WO2009064151A1
Application number: PCT/MX2007/000135
Authority: WO
Inventors: Jaime Tortoriello Garcia; Armando Herrera Arellano; Alejandro ZAMILPA ÁLVAREZ; Jesús Enrique JIMÉNEZ FERRER; Maribel Lucila Herrera Ruiz; Maira Estrella Huerta Reyes; Manasés GONZÁLEZ CORTAZAR
Original assignee: Jaime Tortoriello Garcia
Priority date: 2007-11-12
Filing date: 2007-11-12
Publication date: 2009-05-22

Abstract

The invention relates to pharmaceutical formulations comprising a dry extract of Galphimia glauca for treating anxiety, in which the active agents are compounds known as galphimines A, B and E. The method of obtaining said extract significantly increases the concentration of the active principles (concentration of up to 50 times greater than with previous methods), resulting in a greater anxiolytic effect and enabling pharmaceutical presentation forms which are smaller than those proposed in the prior art, by capsule or coated tablet, which are more effective and user-friendly for patients. The pharmaceutical formulation to which the invention relates has a better anxiolytic effect than extracts disclosed in the prior art, without the side effects of the drugs currently used in anxiety treatments.

Description

PHARMACEUTICAL FORMULATION THAT INCLUDES AN EXTRACT OF Galphimia glauca FOR THE TREATMENT OF ANXIETY

FIELD OF THE INVENTION

"The present invention relates to a pharmaceutical formulation comprising a dry extract of the plant species Galphimia glauca (Malpighiaceae) where said extract is obtained with a process that includes solvents of different polarity. Said formulation is used for the treatment, cure and prevention of anxiety in all its pathological forms.The pharmaceutical formulation, object of the present invention, provides a solution for the treatment of anxiety since said formulation comprises a dry extract with a higher concentration of active compounds that produces a greater anxiolytic effect, which allows pharmaceutical presentations smaller than those proposed in the state of the art by capsule or dragee, which are more effective and accessible for use by patients.These pharmaceutical presentations include capsules, dragees, tablets, which can be made with techniques conventional manufacturing Pharmaceutical

BACKGROUND OF THE INVENTION

Anxiety disorders in their different varieties affect approximately 5% of the general population, two thirds of the total cases are women and usually tends to start between the 3rd. and the 4th. decades of life (30-40 years) (Ayuso JL. 2002, The Primary Care Physician in front of the patient with Generalized Anxiety. Global Health Year II # 6 2002: 2-7). In addition, reports indicate that the prevalence of anxiety worldwide is increasing (from 0.1 to 16.9%) (Demyttenaere K, Bruffaerts R, Posada-Villa J, et al.; WHO World Mental Health Survey Consortium. Prevalence, Severity, and Unmet Need for treatment of Mental Disorders in the World Health Organization World Mental Health Surveys. JAMA 2004; 291: 2581-2590).

It has been reported that extracts of the plant species Galphimia glauca (Malpighiaceae) have anxiolytic properties (Tortoriello, J. and Lozoya, X. Effect of Galphimia glauca methanolic extract on neuropharmacological tests. Plant Med. 1992; 58: 234-236) and they have isolated and identified the active principles referred to collectively as galphiminas (Toscano, RA, Ortega, A., Maldonado, E., Gaviño, R., Lozoya, X., Tortoriello, J. Structure of Galphimine B, Acta Cryst, 1993 ; C49: 774-776; González-Cortázar M., Tortoriello J., Alvarez L. Norofordellanes as spasmolytics, Advances of Structure-Activity Relationships. Planta Med. 2005; 71: 711-716). Galphimins A, B, C, D, E, F, G, H, I and J have been isolated as natural products of the methanolic extract of G. glauca mainly (González-Cortázar M, Tortoriello J, Alvarez L. Nor-secofriedelanes as spasmolytics, advances os structure-activity relationships, Planta Med. 2005; 71: 711-716; Taketa AT, Lozada-Lettuce J., Fragoso-Serrano M., Villareal ML, Pereda-Miranda RJ Nat. Prod. 2004; 67 : 644-649) and, in addition, 7 derivatives of Galphimina-E (González-Cortázar M, Tortoriello J, Alvarez L. Norofriedelanes as spasmolytics, advances os structure-activity relationships. Plant Med. 2005; 71: 711- 716). The chemical structure of said A-J galphimines is shown in Table 1.

The anxiolytic evaluation of these galphimins has been carried out in accordance with high cross-shaped maze behavioral models {plus maze) and, in addition, the effect on spontaneous motor activity has been determined using the open field model (open fleld ) (Herrera-Ruiz M, Jiménez-Ferrer JE, DE Lima TCM, Aviles-Montes D, Pérez-Garcia D, González-Cortázar M, Tortoriello J. Anxiolytic and antidepressant-like activity of a standardized extract from Galphimia glauca. Phytomedicine 2006 ; 13: 23-28; Herrera-Ruiz M, González M, Jiménez E, Zamilpa A, Alvarez L, Ramírez G, Tortoriello J. Anxiolytic effect of natural galphimines from Galphimia glauca and their chemical derivatives. Journal of Natural Products 2006; 69 : 59-61). Table 1. Galphiminas AJ isolated as natural products of the methanolic extract of G. glauca.

Galphimina J R = Ac

The greatest anxiolytic activity has been observed in the compounds Galphimina-B and Galphimina-A (Tortoriello, J. and Ortega, A., Sedative effect of galphimine-B, a nor-dry-triterpenoid from Galphimia glauca. Plant Med. 1993; 59: 398-400; González-Cortazar M., Tortoriello J., Alvarez L. Nor-secofriedelanes as spasmolytics, Advances of Structure-Activity Relationships. Planta Med. 2005; 71: 711-716), however, the abundance of said galphiminas in the extracts of G. glauca is Galphimina-E greater than or equal to Galphimina-B, and Galphimina-B much greater than Galphimina-A (Herrera-Ruiz M, Jiménez-Ferrer JE, DE Lima TCM, Aviles-Montes D , Pérez- Garcia D, González-Cortázar M, Tortoriello J. Anxiolytic and antidepressant-like activity of a standardized extract from Galphimia glauca. Phytomedicine 2006; 13: 23-28; Herrera-Ruiz M, González M, Jiménez E, Zamilpa A , Alvarez L, Ramírez G, Tortoriello J. Anxiolytic effect of natural galphimines from Galphimia glauca and their chemical derivatives. Journal of Natural

Products 2006; 69: 59-61). It has been proposed that extracts of G. glauca of different polar nature such as ethanolic, methanolic and aqueous could be useful in anxiety treatments due to the presence of galphimins A and B in their composition and the absence of demonstrated hepatotoxic, toxicological and genotoxic effects in mouse trials (Herrera-Ruiz M, González M, Jiménez E, Zamilpa A, Alvarez L, Ramírez G, Tortoriello J. Anxiolytic effect of natural galphimines from Galphimia glauca and their chemical derivatives. Journal of Natural Products 2006; 69:59 -61; Aguilar-Santamaría L., Ramírez G., Herrera-Arellano A., Zamilpa A., Jiménez JE, Alonso-Cortés D., Cortes-Gutiérrez EI, Ledesma N, Tortoriello J. Toxicological and cytotoxic evaluation of standardized extracts of Galphimia glauca J. Ethnopharmacology 2007; 109: 35-40). For this reason, a double-blind, randomized clinical trial compared to lorazepam was conducted in patients with a diagnosis of generalized anxiety disorder to assess its efficacy, safety and therapeutic tolerability. The treatment to be evaluated consisted of capsules with dry aqueous extract of G. glauca standardized exclusively in its Galphimina-B content (Herrera-Arellano A, Jiménez-Ferrer E, Zamilpa A, Morales-Valdéz M, García-Valencia Claudia E, Tortoriello J. Efficacy and Tolerability of a standardized herbal product from Galphimia glauca on generalized anxiety disorder A randomized, double-blind clinical trial controlled with Lorazepam. Planta Medica 2007; 73: 713-717). It was shown that after 4 weeks of treatment with the aqueous dry extract of G. glauca, anxiety was reduced in patients by a similar magnitude to that achieved by the lorazepam control, with the obvious advantage of having significantly decreased the side effects caused by control. In the administration of the dry extract of G. glauca, excessive sedation was the most common effect and only occurred in 4 of the 72 patients in the experimental group considered in the study, while in patients treated with lorazepam this side effect was presented. in 16 of them (Herrera-Arellano A, Jiménez-Ferrer E, Zamilpa A, Morales-Valdéz M, García-Valencia Claudia E, Tortoriello J. Efficacy and Tolerability of a standardized herbal product from Galphimia glauca on generalized anxiety disorder. randomized, double-blind clinical trial controlled with Lorazepam. 2007 Medica Plant; 73: 713-717).

The preparation of a medicament comprising an extract of G. glauca has been previously proposed. Patent application MX19990004189 (Tortoriello J, et al) proposes a medicament comprising an extract of G. glauca for the treatment of conditions that involve glutamatergic and / or dopaminergic hyperactivity. However, the medicine proposed in said application has disadvantages, since said extract is only characterized in its chemical composition by the presence of the compound called Galphimine-B. Patent application MX20060008508 (Tortoriello J., et al) claims a pharmaceutical composition comprising at least one of the compounds known as Galphimina-F and Galphimina-B for the treatment of a disease with a spasmogenic dysfunctional condition, as well as for The treatment of an anxiogenic condition. However, said pharmaceutical composition differs substantially from the present invention, because the pharmaceutical composition uses only pure compounds and therefore involves a chemical-pharmaceutical type drug. Meanwhile, the present invention relates to a pharmaceutical composition made from a dry extract that involves an herbal medicine, and therefore, are drugs of a completely different nature. Moreover, in the economic aspect, the pharmaceutical composition made from pure compounds presents another disadvantage, since it involves exhaustive purification processes with a high economic cost due to the use of a more sophisticated infrastructure.

The present invention relates to a pharmaceutical formulation comprising a dry extract of the plant species Galphimia glauca (Malpighiaceae), where the extract has a controlled concentration and much higher than previously reported of the active ingredients Galphimina-B, Galphimina-A and Galphimina-E (Herrera-Ruiz M, Jiménez-Ferrer JE, DE Lima TCM, Aviles-Montes D, Pérez-Garcia D, González-Cortázar M, Tortoriello J. Anxiolytic and antidepressant-like activity of a standardized extract frora Galphimia glauca Phytomedicine

2006; 13: 23-28; Herrera-Ruiz M, González M, Jiménez E, Zamilpa A, Alvarez L, Ramírez G, Tortoriello J. Anxiolytic effect of natural galphimines from Galphimia glauca and their chemical derivatives. Journal of Natural Products 2006; 69: 59-61; Aguilar-Santamaría L., Ramírez G., Herrera-Arellano A., Zamilpa A., Jiménez JE, Alonso-Cortés D., Cortes-Gutiérrez EI, Ledesma N, Tortoriello J. Toxicological and cytotoxic evaluation of standardized extracts of Galphimia glauca . Journal of Ethnopharmacology 2007; 109: 35-40; Herrera-Arellano A, Jiménez-Ferrer E, Zamilpa A, Morales-Valdéz M, García-Valencia Claudia E, Tortoriello J. Effícacy and Tolerability of a standardized herbal product from Galphimia glauca on generalized anxiety disorder. A randomized, double-blind clinical trial controlled with Lorazepam. 2007 Medica Plant; 73: 713-717). Said formulation proposes a notable improvement with respect to the anxiolytic effect shown by other extracts previously disclosed in the state of the art as illustrated in the examples of the present application, and further proposes, oral administration of a lower dose of dry extract of G Glauca, which may produce it in different presentations more accessible and comfortable for use by patients and also achieve a better anxiolytic effect. Additionally, and based on the same clinical study of the aqueous extract of G. glauca, prior to the present application, the registered side effects are much lower than those of the drugs currently used in anti-anxiety therapy, since only excessive sedation It was the most common effect. Furthermore, the aqueous extract of G. glauca begins its therapeutic effect from the first week of administration, unlike the more recent anxiolytics, such as azapirones and serotonin reuptake inhibitors, whose effect is observed from the fourth week onwards. of treatment.

The invention also relates to a method for the preparation of said dry extract, which is obtained by a method of extraction other than extracts already known in the state of the art (aqueous, hydroalcoholic and methanolic) because the method of extraction of the present invention, it is modified in its number of passages and solvents used, since it comprises successive extractions with different organic solvents and mixtures thereof, as well as bipartitions (liquid-liquid separations). With this method Extraction is achieved a significant increase in the concentration of active ingredients up to

50 times higher compared to previous methods.

Additionally, the invention relates to the use of said extract for the manufacture of pharmaceutical formulations in different dosage forms suitable for oral administration which are indicated for the treatment of anxiety.

As a preferred embodiment of the invention, capsules comprising the pharmaceutical formulation of G. glauca are proposed, which administered in patients reduces anxiety from the first week of treatment, and after 4 weeks of treatment significantly reduces anxiety. Additionally, as another preferred embodiment of the present invention, a dragee is proposed, which is coated and comprises the formulation of G. glauca, which administered in patients reduces anxiety from the first week of treatment, and after 4 weeks of Treatment significantly reduces anxiety.

Additionally, as another preferred embodiment of the present invention, a tablet is proposed, which comprises the formulation of G. glauca, which administered to patients reduces anxiety from the first week of treatment, and after 4 weeks of treatment significantly reduces the anxiety.

DESCRIPTION OF THE FIGURES

Figure 1 shows the chromatographic profile of the dry extract of G. glauca object of the invention and shows the retention times of the active ingredients called Galphiminas A, B and E (GA, GB and GE, respectively), as well as of the quercetin used as a reference.

Figure 2 shows the anxiolytic activity of the aqueous (Aq), hydroalcoholic (HAq) extracts and the extract object of the invention (Ext inv) obtained from G. glauca in the model Behavioral cross-shaped maze (plus mazé). Data are shown as percentage of entries to open arms (α) and percentage of time over open arms (a). Diazepam (DZP) is used as a positive control and 1% carboxymethyl cellulose (eme) as a vehicle. Figure 3 compares the spontaneous motor activity of the aqueous (Aq), hydroalcoholic (HAq) extracts and the extract object of the invention (Ext inv) of G. glauca in the open-field behavioral model. The data are shown as total number of crosses (π) and number of times of stretching (m). Diazepam (DZP) is used as a positive control and 1% carboxymethyl cellulose (eme) as a vehicle.

DETAILED DESCRIPTION OF THE INVENTION

The characteristic details of this pharmaceutical formulation indicated for the treatment of anxiety are shown in the following description and in the accompanying examples as well as the preparation of the extracts of the vegetable species used, in the pharmaceutical dosage form, following the same reference signs to indicate the procedures and examples shown.

The pharmaceutical form according to the pharmaceutical formulation object of the present invention is particularly available for oral administration. The dosage forms according to the pharmaceutical formulation object of the present invention can be prepared by conventional manufacturing techniques and methods, among which we can mention:

CAPSULES: The extract of G. glauca, concerning the pharmaceutical formulation object of the present invention, can be mixed with the excipients that allow the proper drying process of the extract either by a spray drying process with hot air or lyophilization. . The dried extract thus obtained is pulverized. The powder has the characteristics of fluidity to be encapsulated in the required dose and hygroscopicity that keeps the pharmaceutical form.

TABLETS: The G. glauca extract, concerning the pharmaceutical formulation object of the present invention, can be mixed with the excipients that allow the proper drying process of the extract either by a spray drying process with hot air or by lyophilization. . The dried extract thus obtained is pulverized and mixed with excipients that allow compaction according to an established dosage.

GRAGEAS: The extract of G. glauca, concerning the pharmaceutical formulation object of the present invention, can be mixed with the excipients that allow the proper drying process of the extract either by a spray drying process with hot air or by lyophilization. . The dry extract thus obtained is pulverized and mixed with excipients that require compaction according to the established dosage. Dragees can have an enteric layer.

In the following examples the method of obtaining the plant material and the preparation of the extract of G. glauca used in the pharmaceutical formulation object of the present invention is presented.

EXAMPLE No. 1

Obtaining the plant material of G. glauca

The plant material referred to in this invention was obtained from seeds of G. glauca. The seeds germinate at room temperature and humidity. The collection of aerial parts of the plant material that may or may not be in bloom (leaves, stems and / or flowers), preferably leaves, takes place between 6 and 12 months, preferably at 8 months of development.

EXAMPLE No. 2

Preparation of the dry extract of G. glauca

The preparation of the dry extract of G. glauca, object of the invention, comprises the steps: a) Drying of the plant material, b) Grinding, c) Deceration, d) Hydroalcoholic extraction, e) Partition in ethyl acetate: water and, f) Evaporation of solvents until the product is dry.

Each of the steps of the preparation method are described below. a) Drying of the plant material The leaves, stems and / or flowers of G. glauca, preferably the leaves, dry in the dark at a temperature of 20 ° C to 35 ° C, maintaining a relative humidity of 15% to 20%, the Drying time is 15 to 20 days in a ventilated area. b) Grinding

The plant material of G. glauca is crushed in a blade mill with 4 to 12 mm mesh, preferably using the 8 mm mesh. c) Deceration i) 10 to 30 kg of crushed plant material of G. glauca (particle size 4 to 12 mm mesh, preferably 8 mm mesh) are introduced into a 300 liter tank, and the plant material is covered with 7 at 10 liters per kg of plant material, preferably 7.5 liters per kg with a mixture of hexane: ethyl acetate solvents in a proportion ranging from 100 to 60%: 0 to 40%, preferably 70% hexane and 30% of ethyl acetate, at a temperature between 20 and 40 ° C, preferably 25 ° C for 24 hours. ii) The solvent mixture is recovered in a rotary evaporator, with a 20-liter ball flask, maintaining the bath temperature in a range of 35 to 55 ⁰ C preferably 45 ⁰ C, with the application of vacuum at a pressure between 240 and 335 mbar, preferably 310 mbar.

The mixture of recovered solvents is used again to repeat the depreciation of the plant material. The operation is performed three times. Decent plant material The resulting dispersion is placed on absorbent paper to allow for evaporation, the removal of the remains of the solvent mixture that was used in the decentration.

d) Hydroalcoholic extraction i) Decent and dry plant material is placed back in the 300 liter tank and the plant material is covered with 7 to 10 liters per kg of plant material, preferably 7.5 liters per kg with a mixture of ethanol: water in a proportion ranging from 70 to 30%: 30 to 70%, preferably 50% ethanol and 50% water, at a temperature between 40 and 60 ° C, preferably 50 ⁰ C for 4 hours to obtain an extract. ii) The extract is passed through a depth filter comprising the following parts: 304 stainless steel press filter, 12 plates 20 x 20 cm and 5 mm deep, 2 holes, a positive displacement pump with head, of 316 stainless steel, with clamp joints. Previously, the filter is prepared with a diatomaceous earth suspension, known as Celite 504 (filter aid). Said suspension is prepared with 200 gr of the filter aid and suspended in 5 liters of the solvent system to be used. Subsequently, it is filtered using a pressure of 0.2 kg / cm ² . The filter dries with a stream of pressurized air until there is no liquid in the filter outlet. iii) The filtered extract is concentrated on a rotary evaporator with a 20-liter ball flask, maintaining the bath temperature in a range of 45 to 65 ° C, preferably 52 ° C with the application of vacuum at a pressure between 175 and 70 mbar , preferably 120 mbar and gradually reducing up to 70 mbar, until an aqueous suspension with an ethanol percentage of less than 10% is obtained. The mixture of recovered solvents is again used to repeat the maceration of the plant material. This stage is done three times.

e) Partition in ethyl acetate: water i) The aqueous suspension with ethanol below 10% is adjusted with water at a concentration of between 200 and 400 mg / ml, preferably 300 mg / ml. Subsequently, acetate is added of ethyl in an amount equal to the resulting final volume and placed in a separatory funnel. The mixture is vigorously stirred and allowed to stand until two phases are formed: an aqueous phase and an organic phase. This operation is carried out as many times as necessary until the organic phase no longer carries more galphimines, which is verified by thin layer chromatography under the following conditions: a sample of the organic phase and a reference sample of Galphimina-B, Galphimina-A and Galphimina-E in a normal phase chromatoplate, with a chloroform elution system: acetone (90:10) and using Ehrlich as developer. The lack of detection of galphimines in the organic chromatoplate sample indicates that the resulting aqueous phase is free of galphimines.

f) Evaporation of solvents until the product is dry i) The organic phase is recovered and concentrated in a rotary evaporator, maintaining the bath temperature in a range of 35 to 55 ⁰ C, preferably at 45 ° C, and vacuum is applied to a pressure between 220 and 260 mbar, preferably at 240 mbar until the volume is minimized, preferably less than 5% of the original volume. ii) The remaining water and traces of the organic solvent present in the extract are removed by a conventional lyophilizer. The lyophilization cycle is carried out for 36 hours under the following conditions: 4 hours with a vacuum of 1050 mbar at -45 ⁰ C with a condenser temperature of -52 ⁰ C, 14 hours with a vacuum of 30 mbar at -15 ° C, 14 hours with a vacuum of 30 mbar at 0 ^o C and, 4 hours with a vacuum of 30 mbar at 20 ° C. Finally, the extract is pulverized.

EXAMPLE No.3

Phytochemical characterization of the dry extract of & glauca For the phytochemical characterization of the dry extract of G. glauca, object of the invention with the quantification of the active substances (galphiminas), a specific high-performance liquid chromatography (HPLC) protocol has been designed . Said protocol is carried out in a HPLC equipment (Waters) connected to a Waters 2695 separation module and a Waters 2996 diode array detector. This chromatography equipment is controlled with a data capture computation program (Empower Pro, Waters). The experimental conditions under which the phytochemical characterization of the dry extract of G. glauca is performed with the quantification of galphimines are: reverse phase column (Alltima HP C18 HL 3u), the mobile phase consists of an isocratic acetonitrile system: water (35: 65), flow of 1.7 ml / min, with a total time of 20 min. Galphimins A, B and E previously isolated in our laboratory and whose identity was corroborated by comparison with the spectroscopic data reported are used as reference standards for determining the concentration of active compounds. The reference galphimins and the extract object of the invention are dissolved in HPLC grade methanol and injected (50 μl). Reference galphimines are injected at concentrations of 50, 100, 200 and 400 μg / ml. The extract object of the invention is analyzed at a concentration of 3 mg / ml. Each sample is run in triplicate. The chromatographic profile is analyzed in a wavelength range between 200 and 400 nm, setting the reading to 220 nm. The calibration curve is obtained by measuring the area under the curve (i? ² > 0.98) of the respective compound, according to the following values shown in Figure 1: Galphimina-A T _R = 11.06 min, average concentration present in the extract : 17 mg / g +2 mg / g; Galphimina-B T _R = 15.93 min, average concentration present in the extract: 50.3 mg / g ± 3 mg / g; Galphimina-E T _R = 21.67 min, concentration present in the extract: 50.3 mg / g ± 3 mg / g. These samples were analyzed in triplicate and from three independent extracts. When comparing these values with those previously reported in the literature for extracts already known in the state of the art such as aqueous, methanolic and ethanolic (Table 2) (Aguilar-Santamaría L., Ramírez G., Herrera-Arellano A., Zamilpa A., Jiménez JE, Alonso-Cortés D., Cortes-Gutiérrez EI, Ledesma N, Tortoriello J. Toxicological and cytotoxic evaluation of standardized extracts of Galphimia glauca. Journal of Ethnopharmacology 2007; 109: 35-40) extract object of the invention has a higher concentration of Galphimine-B, up to 50 times more than the aqueous extract, and almost 3 times more concentration of said galphimine than methanolic and ethanolic extracts. In the same way, the extract object of the invention has a concentration of Galphimine-E greater than the aqueous extract of up to 45 times and 3 and 2 times higher than the methanolic and ethanolic extracts, respectively. Finally, the extract object of the invention has a concentration of Galphimine-A of up to 28 times higher with respect to the aqueous extract and 3 and 2 times more than the ethanol and methanol extracts, respectively.

Figure 1, additionally shows a peak with T _R = 2.06 min belonging to the quercetin compound (with a concentration of 9.5 mg / g) presented in the chromatogram as a reference because it is a compound widely known in the state of The technique.

Table 2. Comparison of the concentration of the active substances galphimins A, B, and E present in dry extracts of G. glauca.

EXAMPLE No. 4

Determination of the anxiolytic activity of the pharmaceutical formulation object of the present invention by means of a cross-shaped high labyrinth model (plus mazé)

The dry extract of G. glauca object of the invention referred to in examples 2 and 3 of the present application, is administered orally in male mice, strain ICR, with 30-35 g weight, at a dose of 100 mg / kg, for three occasions: at 24, 18 and 1 hour before measurement. The anxiolytic effect of this extract shows an improvement with respect to the anxiolytic effect produced by other extracts obtained from the same plant previously used and disseminated (Herrera-Ruiz M, Jiménez-Ferrer JE, DE Lima TCM, Aviles-Montes D, Pérez-Garcia D , González-Cortázar M, Tortoriello J. Anxiolytic and antidepressant-like activity of a standardized extract from Galphimia glauca, Phytomedicine 2006; 13: 23-28; Herrera-Ruiz M, González M, Jiménez E, Zamilpa A, Alvarez L, Ramírez G, Tortoriello J. Anxiolytic effect of natural galphimines from Galphimia glauca and their chemical derivatives, Journal of Natural Products 2006; 69: 59-61; Aguilar-Santamaría L., Ramírez G., Herrera-Arellano A., Zamilpa A., Jiménez JE, Alonso-Cortés D., Cortes-Gutiérrez EI, Ledesma N, Tortoriello J. Toxico logical and cytotoxic evaluation of standardized extracts of Galphimia glauca Journal of Ethnopharmacology 2007; 109: 35-40; Herrera-Arellano A, Jiménez- Ferrer E, Zamilpa A, Morale s-Valdéz M, García- Valencia Claudia E, Tortoriello J. Effícacy and Tolerability of a standardized herbal product from Galphimia glauca on generalized anxiety disorder. A randomized, double-blind clinical trial controlled with Lorazepam. 2007 Medica Plant; 73: 713-717). The comparison of these effects is shown in Figure 2, in which it is observed that the aqueous (Aq) and hydroalcoholic (HAq) extract of G. glauca have considerably lower values of anxiolytic activity, with respect to the extract object of the invention (Ext inv). Data are presented as percentage of entries to open arms (D) and percentage of time over open arms (B). It is used as a diazepam control (DZP) administered at 1 mg / kg ip and as a 1% carboxymethyl cellulose (eme) vehicle. The data analysis is carried out by ANOVA and it is determined that the dry extract, object of the invention and the DZP show significant differences with respect to the vehicle (p <0.05) with a Dunnet post-hoc test. While the aqueous extract (Aq) and the hydroalcoholic extract (HAq) showed no differences with the vehicle. EXAMPLE No. 5

Determination of the effect on the spontaneous motor activity of the pharmaceutical formulation object of the present invention by means of the open field model (hear fíeld)

After the cross-shaped maze test (plus mazé), the effect of the pharmaceutical formulation object of the invention is determined on the spontaneous motor activity of the same batch of mice and with the same controls indicated in example 4. Figure 3 shows the results of the evaluation of the aqueous (Aq), hydroalcoholic (HAq) and dry extracts object of the invention (Ext inv) extracts. The data shown are the total number of crosses (π) and number of times of stretching (u). The total number of crosses refers to the movement of animals from one quadrant to another in the open field device. The number of times of stretching, refers to the incorporations of the animals in the hindquarters (lifts on 2 legs). Statistical analysis showed no significant difference between control and extract treatments when compared to the vehicle. Therefore, the results show that the dry extract object of the invention does not affect the spontaneous motor activity of the experimental animals.

EXAMPLE No. 6

Preparation of capsules comprising the pharmaceutical formulation object of the present invention

To the powdered extract resulting from step f, sub-stage ii of example 2 and chemically characterized in example 3 of the present application, excipients are added in accordance with the following percentages: lactose in a concentration between 12 and 20% solids in solution, preferably 16%; magnesium sterate in a concentration between 1 and 4% solids in solution, preferably 2.5%; surgical talc in a concentration between 0.1 and 1.2% solids in solution preferably at 0.5% and caboxil ^® in a concentration between 0.2 and 1.8% solids in solution preferably at 0.8%. Once the excipients have been added, the extract is placed in a powder mixer with 304 stainless steel tub, with a nominal capacity of 99 liters, and mixed vigorously until a homogeneous product is obtained.

The extract proceeds to be packed in capsules with a capacity of 250 mg. Each capsule shall provide at least the following amounts of active substance: 0.35 mg of Galphimina-B (G-B), 0.34 mg of Galphimina-E (GE) and 0.11 mg of Galphimina-A (GA). ^” These doses are standardized for in accordance with previous studies of the clinical study of the aqueous extract of G. glauca and are administered orally 1 or 2 times a day for an appropriate period of time to observe the anxiolytic effect, which is at least 4 continuous weeks (Herrera- Arellano A, Jiménez-Ferrer E, Zamilpa A, Morales- Valdéz M, García-Valencia Claudia E, Tortoriello J. Efficacy and Tolerability of a standardized herbal product firom Galphimia glauca on generalized anxiety disorder A randomized, double-blind clinical trial controlled with Lorazepam. Planta Medica 2007; 73: 713-717).

Having sufficiently and clearly described my invention, I consider it as a novelty and therefore claim as my exclusive property, what is contained in the following claims.

Claims

1.- A method to obtain a dry extract of Galphimia glauca characterized in that it comprises the stages: a) Drying of the plant material, b) Grinding, c) Deceration, d) Hydroalcoholic extraction, e) Partition in ethyl acetate: water, and f ) Evaporation of solvents until the product is dry.

2, - A method for obtaining a dry extract of Galphimia glauca according to claim No. 1, characterized in that in step a) the plant material is selected from the group consisting of: leaves, stems, flowers and mixtures thereof .

3. A method for obtaining a dry extract of Galphimia glauca according to claims No. 1 to 2, characterized in that the plant material is preferably leaves.

4. A method for obtaining a dry extract of Galphimia glauca according to claims No. 1 to 3, characterized in that in step a) the plant material is dried in darkness at a temperature of 20 ° to 35 ° C.

5, - A method for obtaining a dry extract of Galphimia glauca according to claims No. 1 to 4, characterized in that in step a) the plant material is dried maintaining a relative humidity of 15% to 20%.

6. A method for obtaining a dry extract of Galphimia glauca according to claims No. 1 to 5, characterized in that in step a) the plant material is dried for a period of time between 15 to 20 days in a ventilated area . 7. A method for obtaining a dry extract of Galphimia glauca according to claims No. 1 to 6, characterized in that in step b) the dried plant material is crushed in a blade mill with 4 to 12 mm mesh, preferably using the 8 mm mesh.

8.- A method for obtaining a dry extract of Galphimia glauca according to claims No. 1 to 7, characterized in that in step c) the dried and crushed plant material is decanted with a mixture of hexane solvents: ethyl acetate in a proportion ranging from 100 to 60%: 0 to 40%, preferably 70% hexane and 30% ethyl acetate.

9. A method for obtaining a dry extract of Galphimia glauca according to claims No. 1 to 8, characterized in that step c) is carried out at a temperature between 20 to 40 ° C, preferably 25 ° C. 10. A method for obtaining a dry extract of Galphimia glauca according to claims No. 1 to 9, characterized in that step c) is carried out for 24 hours.

11. A method for obtaining a dry extract of Galphimia glauca according to claims No. 1 to 10, characterized in that step c) is performed three times. 12. A method for obtaining a dry extract of Galphimia glauca according to claims No. 11, characterized in that in step c) the resulting decentered plant material is placed dispersed on absorbent paper to allow evaporation, removal of the residues of the solvent mixture that was used in the depreciation. 13.- A method to obtain a dry extract of Galphimia glauca in accordance with. Claims No. 1 to 12, characterized in that in step d) the decentered material is extracted with a mixture of ethanol-solvent in a proportion ranging from 70 to 30%: 30 to 70%, preferably 50% ethanol and 50% Water.

14. A method for obtaining a dry extract of Galphimia glauca according to claims No. 1 to 13, characterized in that step d) is carried out at a temperature between 40 and 60 ° C, preferably 50 ⁰ C. 15 A method for obtaining a dry extract of Galphimia glauca according to claims No. 1 to 14, characterized in that step d) is carried out for 4 hours.

16. A method for obtaining a dry extract of Galphimia glauca according to claims No. 1 to 15, characterized in that in step d) a filtration of the extract using a Celite 504 filter at a pressure of 0.2 kg / cm ² .

17. A method for obtaining a dry extract of Galphimia glauca according to claims No. 1 to 16, characterized in that in step d) the filtered extract is concentrated in a rotary evaporator. 18. A method for obtaining a dry extract of Galphimia glauca according to claims No. 1 to 17, characterized in that in step d) the bath temperature is maintained in a range of 45 to 65 ° C, preferably 52 ° C.

19. A method for obtaining a dry extract of Galphimia glauca according to claims No. 1 to 18, characterized in that in step d) vacuum is applied at a pressure between 175 and 70 mbar, preferably 120 mbar and gradually reducing to 70 mbar

20. A method for obtaining a dry extract of Galphimia glauca according to claims No. 1 to 19, characterized in that in step d) an aqueous suspension with an ethanol percentage of less than 10% is obtained.

21. A method for obtaining a dry extract of Galphimia glauca according to claims No. 1 to 20, characterized in that in step d) it is performed three times.

22. A method for obtaining a dry extract of Galphimia glauca according to claims No. 1 to 21, characterized in that in step e) the aqueous suspension with ethanol less than 10% is adjusted with water at a concentration of between 200 and 400 mg / ml, preferably at 300 mg / ml. 23. A method for obtaining a dry extract of Galphimia glauca according to claims No. 1 to 22, characterized in that in step e) ethyl acetate is added in an amount equal to the resulting final volume, vigorously stirred and Let stand until two phases are formed: an aqueous phase and an organic phase.

24. A method for obtaining a dry extract of Galphimia glauca according to claims No. 1 to 23, characterized in that in step e) the organic phase is recovered and this operation is carried out as many times as necessary until that the organic phase does not carry more galphimines, which is verified by thin layer chromatography.

25. A method for obtaining a dry extract of Qalphimia glauca according to claims No. 1 to 24, characterized in that in step e) the conditions of thin layer chromatography are: normal phase chromatoplate with an elution system of chloroformoracetone (90:10) and using Ehrlich as a developer. 26.- A method for obtaining a dry extract of Galphimia glauca according to claims No. 1 to 25, characterized in that in step e) the chromatoplac organic sample indicates that the resulting aqueous phase is free of galphimines. 27.- A method for obtaining a dry extract of Galphimia glauca according to claims No. 1 to 26, characterized in that in step f) the recovered organic phase is concentrated in a rotary evaporator.

28.- A method for obtaining a dry extract of Galphimia glauca according to claims No. 1 to 27, characterized in that in step f) the bath temperature is maintained in a range of 35 to 55 ⁰ C, preferably at 45 ⁰ C. 29.- A method for obtaining a dry extract of Galphimia glauca according to claims No. 1 to 28, characterized in that in step f) vacuum is applied at a pressure between 220 and 260 mbar, preferably at 240 mbar .

30.- A method for obtaining a dry extract of Galphimia glauca according to claims No. 1 to 29, characterized in that in step f) the volume is reduced to less than 5% of the original volume. 31. A method for obtaining a dry extract of Galphimia glauca according to claims No. 1 to 30, characterized in that in step f) the remaining water and traces of the organic solvent present in the extract are eliminated by a lyophilization cycle. 36 hours 32.- A method for obtaining a dry extract of Galphimia glauca according to claims No. 1 to 31, characterized in that in step f) the lyophilization cycle is carried out: 4 hours with a vacuum of 1050 mbar at -45 ° C with a condenser temperature of -52 ⁰ C, 14 hours with a vacuum of 30 mbar at -15 ° C, 14 hours with a vacuum of 30 mbar at 0 ^o C and, 4 hours with a vacuum of 30 mbar at 20 ° C.

33.- A method for obtaining a dry extract of Galphimia glauca according to claims No. 1 to 32, characterized in that in step f) the extract is pulverized. 34.- A dry extract of Galphimia glauca obtained by the method as defined in claims No. 1 to 33, characterized in that it comprises a) Galphimine-A in a concentration of 15 to 19 mg per gram of extract; b) Galphimina-B in a concentration of

47.3 to 53.3 mg per gram of extract; and c) Galphimina-E in a concentration of 47.3 mg a

53.3 mg per gram of extract. 35.- A dry extract of Galphimia glauca according to claim No. 34, characterized in that the amount of Galphimine-A present in the extract is preferably of

17 mg / per gram of extract.

36.- A dry extract of Galphimia glauca according to claim No. 34, characterized in that the amount of Galphimine-B present in the extract is preferably 50.3 mg per gram of extract.

37.- A dry extract of Galphimia glauca according to claim No. 34, characterized in that the amount of Galphimina-E present in the extract is preferably of

50.3 mg per gram of extract.

38.- A dry extract of Galphimia glauca according to claims No. 34 to 37 characterized in that it has the profile as shown in Figure 1.

39. A pharmaceutical composition for the treatment of anxiety characterized in that it comprises the dry extract of Galphimia glauca as defined in claims No. 34 to 38 and appropriate excipients.

40. A pharmaceutical composition according to claim No. 39, wherein said composition is a pharmaceutical composition for oral administration.

41. A pharmaceutical composition according to claims No. 39 to 40, wherein the excipients are: lactose in a concentration between 12 and 20% solids in solution, preferably 16%; magnesium sterate in a concentration between 1 and 4% solids in solution, preferably 2.5%; surgical talc at a concentration between 0.1 and

1.2% solids in solution preferably at 0.5% and caboxil ^® in a concentration between 0.2 and

1.8% solids in solution preferably at 0.8%. 42. A pharmaceutical composition according to claims No. 39 to 41, wherein the dosage form is selected from the group consisting of capsules, tablets and enteric layer dragees.

43.- The use of a dry extract of Galphimia glauca as defined in claims No.

34 to 39 to prepare a medication for the treatment of anxiety, wherein said medication is adapted to be orally administrable.

44.- The use of a pharmaceutical composition according to claims 39 to 42 to prepare a medicament for the treatment of anxiety, wherein said medicament is adapted to be orally administrable.

45. The use according to claims No. 43 to 44, wherein the medicament has an oral dosage dosage form selected from the group consisting of capsules, tablets and enteric layer dragees.

46.- The use according to claims No. 43 to 45, wherein the medicament is indicated to be administrable at a dose of 1 or 2 times a day, during an appropriate treatment time to observe the anxiolytic effect. 47.- The use according to claims No. 43 to 46, wherein the treatment time is at least 4 weeks.

48.- The use according to claims 43 to 47 wherein the medicament is adapted to be administrable in such a way as to provide at least an amount of: 0.35 mg of

Galphimina-B, 0.34 mg of Galphimina-E and 0.11 mg of Galphimina-A per dose.