WO2008009084A1

WO2008009084A1 - Preparation and pharmaceutical use of euterpe oleracea (acai) extract compositions

Info

Publication number: WO2008009084A1
Application number: PCT/BR2007/000178
Authority: WO
Inventors: Roberto Soares De Moura
Original assignee: Universidade Do Estado Do Rio De Janeiro
Priority date: 2006-07-18
Filing date: 2007-07-11
Publication date: 2008-01-24
Also published as: ES2705169T3; EP2051721B1; BRPI0604281A; EP2051721A1; EP2051721A4

Abstract

The present invention deal with a process to obtain a decoction from Euterpe oleracea fruits and stones; process to obtain a hydro-alcoholic extract from the decoction; process to obtain a lyophilized and/or spray dried from the hydro- alcoholic extract; pharmaceutical preparations containing the decoction and/or the before mentioned extracts and therapeutic utilization of the preparations in the prevention and treatment of arterial hypertension and other cardiovascular diseases. The extracts induced a significant vasodilator effect in the isolated vascular mesenteric bed of the rats. Capsules and tablets were prepared with the decoctions and lyophilized and or spray dryer of the extracts.

Description

PREPARATION AND PHARMACEUTICAL USE OF EUTERPE OLERACEA (ACAI) EXTRACT COMPOSITIONS

I) Field of Invention

The present invention deal with products that have anti-hypertensive properties, process to obtain products from plants belonging to Palmae family, more specifically to Euterpe oleracea specie, application of those products, process to obtain those products, more specifically a process to obtain a decoction from fruit of this plant, more specifically a process to obtain a decoction from fruits and/or stone of the fruit, more specifically a process to obtain a hydro-alcoholic extracts from those decoctions, more specifically, a process to obtain a lyophilized of those extracts, more specifically obtainment of pharmaceutical preparations and therapeutic indications in the treatment of vasoespastics and ischemic diseases and particularly arterial hypertension.

II) Invention Antecedents

Ischemic diseases are a large problem in medicine particularly in cardiology where we can found vasospasm causing various circulatory problems as ischemic cardiopathy and vascular peripheral vascular diseases.

Therefore search for new medicines that can reduce the contraction of vascular smooth muscle is a permanent goal in pharmacology.

Arterial hypertension is a disease with high prevalence among adult population and induces many deleterious effects in hypertensive patients, including cardiac, kidney and cerebral dysfunctions. Arterial hypertension is at the moment one of the largest causes of death. Therefore, pharmacological treatment that reduces the high level of arterial blood pressure and the cardiovascular complications from arterial hypertension is helpful for the patient and supported by health public agency. Usually the pharmacological treatment of hypertension is obtained with use of diuretics, beta blocking agents, inhibitors of the renin-angiotensin system, inhibitors of the sympathetic system and vasodilators compounds. Ill) Summary of the Invention.

The present invention refers to a process to obtain products that have vasodilator and anti-hypertensive properties. Refer also of those products that are obtained from plants belonging to Palmae family, more specifically Euterpe oleracea specie, in particularly, to a process to obtain products obtained from decoctions obtained from the fruits and/or stones of the fruits. In particular a process for obtainment of the before mentioned products that means submit the fruits or the stones of the fruits of Euterpe oleracea, to an extraction to obtain a lyophilized and/or spray dried of those products.

More particularly, the present invention, refer the to a process to obtain products that have vasodilator and anti-hypertensive activity, that include utilization of the hole fruit or only the stone of the fruit, process to obtain a decoction from the whole fruit or only the stone of the fruit, extraction of the decoction with solvents, particularly solvents physiologically acceptable as ethanol, and process to obtain a lyophilized and/or spray dried of the pharmacological active products.

More particularly, refer also the present invention, methods to obtain products with vasodilator and anti-hypertensive properties as a lyophilized or spray dried, that means to submit the fruit or the stone of the fruit to an extraction with o physiological solvent, for instance water, as a decoction for 3 to 30 minutes. Also the present invention refers to o process of extraction of the decoction with ethanol for 5 to 15 days and posterior filtration and evaporation of the ethanol and also obtention of a lyophilized and/or spray dried with pharmacological activities. Furthermore, the present invention, as already mention above, refer to the before mentioned products per se and also the utilization of the before mentioned products and medicines containing the before mentioned products with vasodilator and anti-hypertensive activity. IV) Condensed description of the methodology used. The present invention is supported by scientific data obtained in chemical and pharmacological experimentations. Below we give a brief view of the investigative process that supports the invention.

IV.A) Method to obtain the liophyled and/or the spray dried of the hvdro- alcoholic extract of the fruit or stone of the fruits of Euterpe oleracea. Euterpe oleracea fruits or only the stone of the fruits were washed in tap water and boiled in distilled water for 5 to 10 minutes and then minced and/or grinded, and then boiled and then extracted with ethanol (v: v) and macerated for 7 to 10 days and then the extract was filtered and the liquid phase is concentrated in a low-pressure rotator evaporator at approximately 4O⁰C and then lyophilize or spay dried and kept in low temperature until the day of use.. IV. B) Pharmacodynamic and pharmacotechnical methods.

The pharmacological activities of the products obtained from Euterpe oleracea was assessed by pharmacological tests that assessed the vasodilator effect in rats. The pharmacotechnical method refers the way to obtain capsules or tablets containing the lyophilized and/or the spray dried of the extracts obtained from Euterpe oleracea fruits or stone of the fruits. V.) Detailed description of the Invention.

In the ambit of the present invention, the fruits of Euterpe oleracea specie, before been submitted to the process of extraction, according to the invention, if not utilized after the harvest, can be stored for long periods at the temperature from +4 to -2O⁰C.

V A) Method to obtain the dried residue. Ivophilized or spray dried of the hydro alcoholic extract of the decoction of the whole fruits or decoction of the stone of the fruits of Euteme oleracea. The stones of the fruit of Euterpe oleracea are obtained by elimination of the skin of the whole fruit. The fruits or the stones are washed and boiled in distilled water for 3 to 9 minutes to obtain the decoction of the whole fruits or from the stones. After boiling the decoction are minced and/or grinded, and then boiled for another 5 minutes e extracted with ethanol 95% in a proportion that varies from 1 :0.5 to 2:10, for instance 1:1. This mixture is again minced and/or grinded, and then macerated for a certain period of time (3 hours to 15 days) for instance 6 days at 4° C or at room temperature for instance 25° C under intermittent agitation. At the end of the maceration time the extract is filtered in a sieve with 0.1 to 1.0-mm pore, for instance 0.2 mm, being also filtered through gauze and finally filtered through a paper filter, for instance Whartman n.1. The semi-solid phase can be re-extracted on the same conditions already described for 1 to 3 times, for instance 2 times. The first liquid phase is kept inside a refrigerator at 4° C. The subsequent liquid phases are added to the first one. The liquid phase is concentrated in a low-pressure evaporator at a temperature of 35 to 65 ° C for instance 50° C and then lyophilized or spray dried. V. B) Pharmacological and Pharmacotechnique Methods.

The pharmacological activity of the products obtained from the fruits and/or the stones of Euterpe oleracea were assessed by pharmacological test in rats. The vasodilator effect was assessed in the isolated mesenteric vascular bed of the rat. The vasodilator effect of the extracts was assessed in the vascular mesenteric bed of the rat. Briefly, the rats were killed and the mesenteric artery was cannulated and perfused with Krebs solution with a pulsatile pump 4 ml_ per minute. The perfusion pressure was recorded. The vasodilator effect of the extract as assessed as reduction of the perfusion pressure induced by intra-arterial injections of the extract, during infusion of norepinephrine. The pharmacotechnique method refers to the method to obtain capsules or tablets containing the lyophilized or spray dried residue of the products obtained from Euterpe oleracea. Vl.) General Observation.

The specific procedure described in item V. A to V. B above, were described not with the scope to limit, but wit the scope to illustrate the possibilities of realization of the present invention, with ambit is limited only by the claim annex.

VIh Illustrative examples of the Invention.

Below are examples with the objective to illustrate and not to limit the present invention, which purpose, as mention above, has its delimitation's in the claim annex.

VH.1) Method to obtain the dried residue. Ivophilized and/or spray dried of the fruits of Euterpe oleracea decoction.

Approximately 200 g of Euterpe oleracea fruits were washed in tap water and boiled in 400 ml. of distilled water during 5 minutes. After boiling the fruits were strongly minced and/or grinded and then 400 ml_ of ethanol was added to the decoction and again minced. This extract was macerate for 6 days and being also shaken for at least 1 hour per day. After maceration the decoction was filtered through a sieve with 0.2 mm pores and then filtered in gauze and then through a filter paper Whatman n.1. The liquid phase, obtained after filtration was concentrated under low pressure evaporator at 45° C, lyophilized and/or spray dried and kept at -20° C, until the day of use. VII.2) Method to obtain the dried residue. Ivophilized and/or spray dried of the stone of Euterpe oleracea decoction. Approximately 200 g of Euterpe oleracea fruits were washed in tap water and the stone was separated from the skin of the fruits. The stone was boiled in 400 mL of distilled water during 5 minutes. After boiling the stones were strongly minced and/or grinded and then 400 mL of ethanol was added to the decoction and again minced. This extract was macerate for 6 days and being also shaken for at least 1 hour per day. After maceration the decoction was filtered through a sieve with 0.2 mm pores and then filtered in gauze and then through a filter paper Whatman n.1. The liquid phase, obtained after filtration was concentrated under low pressure evaporator at 45° C, lyophilized and/or spray dried and kept at -20° C₁ until the day of use. V1I.3). Example of biological tests performed with the products of the invention. The vasodilator effect of the products were assesses in the isolated vascular mesenteric bed of the rat precontracted with norepinephrine. VII.3.1) Isolated Mesenteric Vascular Bed

The rat superior mesenteric vascular bed (MVB) was isolated according to McGregor (1965). Briefly, male Wistar rats were killed with inhaled CO2 and the superior MVB was cannulated and perfused at a flow rate of 4ml min^"1 with a physiological salt solution (PSS) by a pulsatile pump (Lifecare Model 4, Abbott' Shaw). The PSS had the following composition (mM): NaCI 118, KCI 4.7, CaCI₂ 2.5, MgSO₄ 1.2, KH₂PO₄ 1.2, NaHCO₃ 25, EDTA 0.02, and glucose 11. The PSS (37°C) was bubbled with 95%O₂/5% CO₂. Perfusion pressure (PP) was measured with a transducer connected to a preamplifier and chart recorded. The lyophilizeds were either dissolved in PSS and administered as bolus injections directly into the perfusion stream (volume < 300 μl). The preparations were left to equilibrate for 30 minutes, and then injections of 120 μmol KCI were administered every 10 minutes until consistent responses were obtained. Lyophilized of the hydroacoholic extract (1 - 30 μg) was injected in bolus after the PP had been elevated (80-110 mm Hg) with norepinephrine (NE; 6-30 μM) added to the perfusion fluid. When the pressor effect of NE reached a plateau, acetylcholine (ACh; 10 pmol) and nitroglycerin (NG; 1 nmol) were injected to test the endothelium-dependent and -independent responses before dose-response curves to lyophilized were obtained. The vasodilator effect of the extracts was expressed as a percent decrease in relation to the pressor effect of NE. The vasodilator effect of the lyophilized obtained from the fruit and from the stone is showed in Figures 1 and 2. Vl II) Pharmacotechnical aspects of the preparation of capsules and tablets containing the Ivophilized obtained from the hvdroalcoholic extract obtained from decoctions of fruit and stone of Euterpe oleracea.

The capsules and/or tablets containing 100 to 1000 mg of lyophilized of the hydroalcoholic extract obtained from decoctions of fruit and stone of Euterpe oleracea were obtained according to the usual Pharmacotechnical procedures. The capsules were obtained in order to contain 10 to 1000 mg, for instance 500 mg of the lyophilized plus cornstarch and colloidal silicon dioxide. Each capsule could have the following composition:

Lyophilized 500 mg 49.9%

Corn starch 250 mg 49.9%

Coloidal silicon dioxide 0.5 mg 0.2%

Total 750.5 mg 100%

Cornstarch was added to complete the total mass of the capsule to approximately 750 mg. Colloidal silicon dioxide was used to adsorb humidity and to facilitate the preparation of the capsules.

Claims

PROCESS TO OBTAIN DECOCTIONS OF SKINS AND STONE OF THE FRUIT OF EUTERPE OLERACEA (AQAI), PROCESS TO OBTAIN A HYDRO- ALCOHOLIC EXTRACT FROM THE DECOCTIONS, PROCESS TO OBTAIN A LYOPHILIZED AND /OR SPRAY DRIED FROM THE HYDRO-ALCOHOLIC EXTRACTS; PROCESS TO OBTAIN PHARMACEUTICAL PREPARATIONS CONTAINING THE LYOPHILIZED AND/OR SPAY DRIED, AND THERAPEUTIC INDICATIONS OF THE PREPARATIONS AS VASODILATORS AND ON THE PREVENTION AND TREATMENT VASOSPASTIC ISCHEMICAL SYNDROMES AND ARTERIAL HYPERTENSION.

1 ) Process to obtain decoction of fruits of Euterpe oleracea characterized by the following steps: a) Wash the fruits in tap water; b) Submit the fruits of step (a) to a process of extraction in boiling water for 3 to 10 minutes to obtain the decoction; c) Grind the decoction of step (b) with a mince during 3 to 10 minutes; d) Boil the decoction during 3 to 10 minutes.

2) Process according to claim 1 characterized by the reason that the extraction of step (b) takes 5 minutes, mincing the decoction (c) takes 5 minutes and the decoction is boiled (d) during 5 minutes.

3) Process to obtain to obtain a hydro-alcoholic extract from the decoction, characterized by the reason that in addition the decoction obtained in step (d) of claim 1 is extracted with the solvent ethanol in the proportion of 1 :1 , v:v and minced strongly during 5 minutes a then macerated for of 6 hours to 10 days at room temperature or at 4⁰C, obtaining, therefore, the hydro-alcoholic extract of the decoction. 4) Process according to claim 3, by the reason the time of maceration is 6 days. 5) Process according to claim 4 characterized by the reason that the macerated is filtered throughout a sieve with 1.0-mm pores and subsequently in filter paper.

6) Process according to claim 5 characterized by the reason that the liquid phase is concentrated in a rotator evaporator at low pressure and at 40 to 60°

C.

7) Process according to claim 6 characterized by the reason that the alcohol free liquid phase after concentration is submit a process to obtain a lyophilized and/or a spray dried, leading to the obtainment of a lyophilized or a spray dried from the hydro-alcoholic extract with pharmacological activity.

8) Pharmaceutical preparations characterized by the reason that contain 10 to 1000 mg of lyophilized and/or spray dried obtained from the hydro-alcoholic extract obtained from the decoction of fruits of Euterpe oleracea and also other inactive pharmacological components. 9) Therapeutic indications of the pharmaceutical preparations pointed in claim 8, characterized by the reason of being indicated in the prevention and treatment of spastic diseases, arterial hypertension and diseases induced by arterial hypertension.

10) Therapeutic indications according to claim 9 by the reason that the pharmaceutical preparation have human and veterinary applications.

11) Process to obtain decoction of stones of the fruits of Euterpe oleracea characterized by the following steps: a) Wash the fruits in tap water; b) Remove the skins of the fruits of step (a) to obtain the stones of the fruits; c) Submit the stones of step (b) to a process of extraction in boiling water for 3 to 10 minutes to obtain the decoction of the stones; d) Grind the decoction of step (c) during 3 to 5 minutes; e) Boil the decoction during 3 to 10 minutes.

12) Process according to claim 11 characterized by the reason that the extraction of step (b) takes 5 minutes, mincing the decoction (c) takes 5 minutes and the decoction is boiled (d) during 5 minutes. 13) Process to obtain a hydro-alcoholic extract from the decoction of stones of the fruits of Euterpe oleracea, characterized by the reason that in addition the decoction obtained in step (d) of claim 11 is extracted with the solvent ethanol in the proportion of 1:1, v:v and grinded and minced strongly during 5 minutes a then macerated for of 6 hours to 10 days at room temperature or at 4⁰C, obtaining, therefore, the hydro-alcoholic extract of the decoction.

14) Process according to claim 3, by the reason the time of maceration is 6 days.

15) Process according to claim 14 characterized by the reason that the macerated is filtered throughout a sieve with 1.0-mm pores and subsequently in filter paper.

16) Process according to claim 15 characterized by the reason that the liquid phase obtained after filtration is concentrated in a rotator evaporator at low pressure and at 40 to 60° C. 17) Process according to claim 16 characterized by the reason that the ethanol free liquid phase after concentration is submit a process to obtain a lyophilized and/or a spray dried, leading to the obtainment of a lyophilized or a spray dried from the hydro-alcoholic extract with pharmacological activity.

18) Pharmaceutical preparations characterized by the reason that contain 10 to 1000 mg of lyophilized and/or spray dried obtained from the hydro-alcoholic extract obtained from the decoction of stone of the fruits of Euterpe oleracea and also other inactive pharmacological components.

19) Therapeutic indications of the pharmaceutical preparations pointed in claim 18, characterized by the reason of being indicated in the prevention and treatment of spastic diseases, arterial hypertension and diseases induced by arterial hypertension.

20) Therapeutic indications according to claim 19 by the reason that the pharmaceutical preparation have human and veterinary applications.