V.) DETAILED DESCRIPTION OF THE INVENTION

[0017] In the ambit of the present invention, the fruits of Vitis labrusca and Vitis vinifera species, 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 −20° C.

V.A) METHOD TO OBTAIN THE GRAPE-SKIN DECOCTION

[0018] Fruits of Vitis labrusca or Vitis vinifera are washed in water, and the skin is separated from the pulp. The skins are washed in water e later put inside of neutral glass or stainless steel recipient in the proportion of 1 to 100 g of skin to 100 ml of water, for instance 25 g /100 ml and boiled for 3 to 60 minutes, for instance about 5 minutes. After the time of boiling, the decoction is left to cool to 20 to 90° C., for instance 75° C. and minced. The minced decoction was macerated for a certain period of time (1 hour to 30 days) for instance 6 hours inside a refrigerate at 4° C. or at room temperature for instance 25° C. under agitation. The decoction 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 Whartman n.1. The liquid phase is concentrated in a low-pressure evaporator at a temperature of 30 to 60° C. for instance 40° C. and then lyophilized and kept under −4 to −70° C.

V.B) METHOD TO OBTAIN THE GRAPE-SKIN HYDRO-ALCOHOLIC EXTRACT FROM THE DECOCTION

[0019] The grape-skin decoction, after been minced short after boiling, is extracted with ethanol 95% in a proportion of decoction/ethanol (v:v) of 1:0.5 to 1:10, for instance 1:1. That mixture is minced and macerated for a certain period of time of 3 hours to 30 days, for instance 6 hours and kept inside a refrigerator at 4° C. or at room temperature at 25° C. and shaken. At the end of the maceration period it is filtered through a sieve with 0.1 to 1 mm pores, for instance 0.2 mm, being also filtered through gauze and finally filtered through a filter paper, Whartman n.1. The semi-solid phase can be extracted again in the same conditions as described above for 1 or 3 times, for instance 2 times. The liquid phase of the first extraction is kept inside a refrigerator at 4° C. and then added to the liquid phase of the other extractions. The final liquid phase is concentrated under low pressure evaporator at a temperature of 35 to 65° C., for instance 40° C. e then lyophilized and kept at −4° C. to −70° C., for instance −20° C.

V.C) METHOD TO OBTAIN THE GRAPE-SKIN HYDRO-ALCOHOLIC-ACETATE OF ETHYL EXTRACT FROM THE DECOCTION

[0020] The decoction, after been minced after boiling, is extracted with a mixture of decoction/ethanol/ethyl acetate (v:v:v) of variable proportion for instance 1:1:1. That mixture is minced and macerated for a certain period of time of 3 hours to 30 days, for instance 6 hours and kept inside a refrigerator at 4° C. or at room temperature at 25° C. and shaken. At the end of the maceration period the extract is filtered through a filter paper Whartman n.1 and the liquid phase kept inside a refrigerator at 4° C. and the semi-solid phase can be again extracted at the same condition as described above for one or three times, for instance two times. The liquid phase of the first extraction is kept inside a refrigerator at 4° C. and then added to the liquid phase of the other extractions. The final liquid phase is concentrated under low pressure evaporator at a temperature of 35 to 65° C., for instance 40° C. e then lyophilized and kept at 4 C to −70° C., for instance −20° C.

VI.) GENERAL OBSERVATION

[0021] The specific procedure described in item V.A to V.C 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.

VII.) PHARMACODYNAMICAL AND PHARMACOTECHNICAL METHODS

[0022] The pharmacological activities of the various products obtained from the Vitis labrusca and Vitis vinifera skins were assessed by pharmacodynamic methods that study the anti-hypertensive activity of the products in the following models of experimental hypertension: DOCA-salt hypertension, spontaneous hypertensive rats (SHR) and hypertension induced by inhibition of nitric oxide synthase. The pharmacotechnical method refer to the method to obtain capsules containing the lyophilized residue of the products obtained from Vitis labrusca and Vitis vinifera skins.

VIII) ILLUSTRATIVE EXAMPLES OF THE INVENTION

[0023] 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.

VIII.A) METHOD TO OBTAIN THE LYOPHILIZED OF THE VITIS LABRUSCA GRAPE-SKIN DECEPTION

[0024] Approximately 2 000 g of Vitis labrusca fruits were washed in tap water. The skins were isolated from the pulps and washed in tap water during approximately 3 minutes. The skin (1000 g) were boiled in 4000 mL of distilled water for 5 minutes. After boiling, while the decoction was still warm, the decoction was minced at approximately 80° C. and left for maceration for 6 hours under shaking. 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 is concentrated under low pressure evaporator at 45° C., lyophilized and kept at −20° C., until the day of use.

VIII.B) METHOD TO OBTAIN THE LYOPHILIZED OF THE VITIS VINIFERA GRAPE-SKIN DECEPTION

[0025] Approximately 2 000 g of Vitis vinifera fruits were washed in tap water. The skins were isolated from the pulps and washed in tap water during approximately 3 minutes. The skin (1000 g) were boiled in 4000 mL of distilled water for 5 minutes. After boiling, while the decoction was still warm, the decoction was minced at approximately 80° C. and left for maceration for 6 hours under shaking. 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 is concentrated under low-pressure evaporator at 45° C., lyophilized and kept at −20° C., until the day of use.

VIII.C) METHOD TO OBTAIN THE LYOPHILIZED OF THE HYDRO-ALCOHOLIC EXTRACT OBTAINED FROM VITIS LABRUSCA GRAPE-SKIN DECOTION

[0026] Approximately 1 000 g of Vitis labrusca fruits were washed in tap water. The skins were isolated from the pulps and washed in tap water during approximately 3 minutes. The skin (500 g) were boiled in 2 L of distilled water for 5 minutes. After boiling, the decoction was minced. Two liter of ethanol was added to the decoction and minced. The extracted was left macerating for 6 hours and filtered through a sieve, pores 0.2-mm and then filtered through a filter paper Whartman n.1. The liquid phase, obtained after filtration was is concentrated under low-pressure evaporator at 45° C., to evaporate the ethanol and then lyophilized to obtain the lyophilized of the hydro-alcoholic extract of the Vitis labrusca decoction.

VIII.D) METHOD TO OBTAIN THE LYOPHILIZED OF THE HYDRO-ALCOHOLIC EXTRACT OBTAINED FROM VITIS VINIFERA GRAPE-SKIN DECEPTION

[0027] Approximately 1 000 g of Vitis vinifera fruits were washed in tap water. The skins were isolated from the pulps and washed in tap water during approximately 3 minutes. The skin (500g) were boiled in 2 L of distilled water for 5 minutes. After boiling, the decoction was minced. Two liters of ethanol was added to the decoction and minced. The extracted was left macerating for 6 hours and filtered through a sieve, pores 0.2-mm and then filtered in a filter paper Whartman n.1. The liquid phase, obtained after filtration was is concentrated under low-pressure evaporator at 45° C., to evaporate the ethanol and then lyophilized to obtain the lyophilized of the hydro-alcoholic extract of the Vitis vinifera decoction.

VIII.E) METHOD TO OBTAIN THE LYOPHILIZED OF THE HYDRO-ALCOHOLIC-ETHYL ACETATE EXTRACT OBTAINED FROM VITIS LABRUSCA GRAPE-SKIN DECEPTION

[0028] Approximately 1 000 g of Vitis labrusca fruits were washed in tap water. The skin (500g) were isolated from the pulps and washed in tap water during approximately 3 minutes. The skins were boiled in 2 L of distilled water for 5 minutes. After boiling, the decoction was minced. One liter of ethanol plus one liter of ethyl acetate were added to the decoction and minced. The extracted was left macerating for 6 hours and filtered through a sieve, pores 0.2-mm and then filtered in a filter paper Whartman n.1. The liquid phase, obtained after filtration was is concentrated under low-pressure evaporator at 45° C., to evaporate the ethanol and then lyophilized to obtain the lyophilized of the hydro-alcoholic extract of the Vitis labrusca decoction.

VIII.F) METHOD TO OBTAIN THE LYOPHILIZED OF THE HYDRO-ALCOHOLIC-ETHYL ACETATE EXTRACT OBTAINED FROM VITIS VINIFERA GRAPE-SKIN DECEPTION

[0029] Approximately 1 000 g of Vitis vinifera fruits were washed in tap water. The skins were isolated from the pulps and washed in tap water during approximately 3 minutes. The skins were boiled in 2 L of distilled water for 5 minutes. After boiling, the decoction was minced. One liter of ethanol plus one liter of ethyl acetate were added to the decoction and minced. The extracted was left macerating for 6 hours and filtered through a sieve, pores 0.2-mm and then filtered in a filter paper Whartman n.1. The liquid phase, obtained after filtration was is concentrated under low-pressure evaporator at 45° C., to evaporate the ethanol and then lyophilized to obtain the lyophilized of the hydro-alcoholic extract of the Vitis vinifera decoction.

IX). EXAMPLE OF BIOLOGICAL TESTS PERFORMED WITH THE PRODUCTS OF THE INVENTION

[0030] The anti-hypertensive activity of lyophilized from various products was access by testing its efficacy of the lyophilized to reduce the levels of experimental arterial hypertension and to reduce the development of hypertension in rats. The anti-hypertensive activity was accessed in adult male Wistar rats, spontaneous hypertensive or made hypertensive by the following methods: nitric oxide inhibition by use of an analogue of L-arginine, that is, L-NAME, and subcutaneous injection of DOCA followed by orally administration of saline in uninephrectomized rats;

[0031] Arterial blood pressure was measured in the tail of rats by a noninvasive method while the rats were awake, using a cuff and a sensor c connected to equipment manufactured by Letica-Barcelona-Spain. The lyophilized was administrated orally, in the drinking water, so that the rats were treated with the products continuously during the period of treatment. Arterial pressure was measured three times per week before and during the treatment with the lyophilized. The values of arterial blood pressure were compared using Student's test and the differences were considered significantly when p<0.05.

IX.A) EFFECTS OF THE LYOPHILIZED OBTAINED FROM THE DECOCTION OF VITIS LABRUSCA SKINS IN THE ARTERIAL HYPERTENSION INDUCED BY INHIBITION OF NITRIC OXIDE SYNTHESIS IN RATS

[0032] Adult, male Wistar rats (250-350 g, n=10) were kept cage with no more than 4 animals por cage. Mean arterial blood pressure was measured noninvasively, by Letica (Barcelona) equipment. A cuff and a sensor around the tail of the animal were connected to the equipment. The cuff is inflated automatically in order the measure the mean arterial blood pressure.

[0033] After a period of adaptation the experimental conditions, for measurement the mean arterial blood pressure, the pharmacodynamical test was started. During the adaptation period, body weight was estimated three times a week, the animals received tap water and food at libitum, and the daily intake of water was estimated.

[0034] The animals were divided in two groups of 5 rats. One group (control) was treated orally with L-NAME, 50 mg/kg/day, diluted in the drinking water. The other group was also treated with L-NAME, 50 mg/kg/day plus 100 mg/kg/day of the lyophilized of the decoction of Vitis labrusca in the drinking water. FIG. 1 show the anti-hypertensive effect of the lyophilized of the decoction of Vitis labrusca in this particular experiment.

IX.B) EFFECTS OF THE LYOPHILIZED OF THE HYDRO-ALCOHOLIC EXTRACT OBTAINED FROM THE DECOCTION OF VITIS LABRUSCA SKINS IN THE ARTERIAL HYPERTENSION INDUCED BY INHIBITION OF NITRIC OXIDE SYNTHESIS IN RATS

[0035] Adult, male Wistar rats (250-350 g, n=12) were kept cage with no more than 4 animals por cage. Mean arterial blood pressure was measured noninvasively, by Letica (Barcelona) equipment. A cuff and a sensor around the tail of the animal were connected to the equipment. The cuff is inflated automatically in order the measure the mean arterial blood pressure.

[0036] After a period of adaptation the experimental conditions, for measurement the mean arterial blood pressure, the pharmacodinamical test was started. During the adaptation period, body weight was estimated three times a week, the animals received tap water and food at libitum, and the daily intake of water was estimated.

[0037] The animals were divided in two groups of 6 rats. One group (control) was treated orally with L-NAME, 50 mg/kg/day, diluted in the drinking water. The other group was also treated with L-NAME, 50 mg/kg/day plus 100 mg/kg/day of the lyophilized of the hydro-alcoholic extract of decoction of Vitis labrusca in the drinking water, five days after the beginning of treatment with L-NAME. FIG. 2 show the anti-hypertensive effect of the lyophilized of the hydro-alcoholic extract obtained from the decoction of Vitis labrusca in this particular experiment.

IX.C) EFFECTS OF THE LYOPHILIZED OF THE HYDRO-ALCOHOLIC-ACETATE OF ETHYL EXTRACT OBTAINED FROM THE DECOCTION OF VITIS LABRUSCA SKINS IN THE ARTERIAL HYPERTENSION INDUCED BY INHIBITION OF NITRIC OXIDE SYNTHESIS IN RATS

[0038] Adult, male Wistar rats (250-350 g, n=12) were kept cage with no more than 4 animals por cage. Mean arterial blood pressure was measured noninvasively, by Letica (Barcelona) equipment. A cuff and a sensor around the tail of the animal were connected to the equipment. The cuff is inflated automatically in order the measure the mean arterial blood pressure.

[0039] During the adaptation period, the animal received food and water “as libitum” and the daily intake of water was estimated. Body weight was estimated three times a week. After the levels of basal pressure were obtained, the rats were treated with L-NAME 70 mg/kg/day in the drinking water. Once the arterial pressure reached elevated level, the animal was treated with 100 mg/kg/day of the lyophilized of the hydro-alcoholic-ethyl acetate plus L-NAME. As can been observed in FIG. 3 , the extract induced a significant anti-hypertensive effect in this particular experiment.

IX.D) EFFECTS OF THE LYOPHILIZED OF THE HYDRO-ALCOHOLIC-ETHYL ACETATE EXTRACT OBTAINED FROM THE DECOCTION OF VITIS LABRUSCA SKINS IN RATS WITH SPONTANIOUS HYPERTENSION

[0040] The antihypertensive activity of hydro-alcoholic-ethyl acetate extract obtained from the decoction of Vitis Labrusca was accessed in five spontaneous hypertensive rats weighting 250 to 350 g. Before the beginning of the treatment with the extract the arterial pressure was measured during two weeks, when after that period, the animals were treated orally with lyophilized of the extract 100 mg/kg/day. As showed in FIG. 4 , oral treatment with the extract induced a significant anti-hypertensive response in this particular kind of experimental hypertension.

IX.E) EFFECTS OF THE LYOPHILIZED OF THE HYDRO-ALCOHOLIC EXTRACT OBTAINED FROM THE DECOCTION OF VITIS LABRUSCA SKINS IN RATS WITH DOCA-SALT HYPERTENSION

[0041] Twelve Wistar male rats, weighting 250 to 350 g were divided in two groups, each group kept in different cages. All rats were were uninephrectomized. Arterial pressure was measured non invasively using a Letica (Barcelona) equipment, that include a cuff, a sensor, put around the rat tail and connected to the equipment that inflate automatically the cuff and therefore measure the level of mean arterial pressure.

[0042] Seven days after the nephrectomy, the animal is adapted to the pressure measurement procedure and received food and water “ad libitun” and body weight was estimated three times a week.

[0043] After the period of adaptation, the animals were treated subcutaneously with 25 mg/kg/week and received saline as drinking water. Arterial pressure was measured three times a week. After the beginning of the treatment, the arterial blood pressure started to elevate. Sixteen days after the beginning of treatment, when the arterial blood pressure had reached a high level, 6 rats were treated orally with the extract 100 mg/kg/day. The other group of 6 rats were treated with only DOCA plus saline. As showed in FIG. 5 , the extract induced a significant reduction of the mean arterial blood pressure in hypertensive rats.

IX.F) EFFECTS OF THE LYOPHILIZED OBTAINED FROM THE DECOCTION OF VITIS VINIFERA SKINS IN THE ARTERIAL HYPERTENSION INDUCED BY INHIBITION OF NITRIC OXIDE SYNTHESIS IN RATS

[0044] Adult, male Wistar rats (250-350 g, n=11) were kept cage with no more than 4 animals por cage. Mean arterial blood pressure was measured noninvasively, by Letica (Barcelona) equipment. A cuff and a sensor placed around the tail of the animal were connected to the equipment. The cuff is inflated automatically in order the measure the mean arterial blood pressure.

[0045] The animals were submitted to a period of adaptation of the experimental conditions, for measurement the mean arterial blood pressure. During the adaptation period, the animals received tap water and food “ad libitum”, and the daily intake of water was estimated. Body weight was estimated three times a week.

[0046] After the basal pressure levels are obtained, one group (control, n=5) was treated orally with 70 mg/kg/day L-NAME in drinking water. The other group (n=6) was also treated with the same dose of L-NAME plus 100 mg/kg/day of lyophilized hydro-alcoholic extract obtained from the decoction of Vitis vinifera skin in the drinking water. As shown in FIG. 6 , the extract induced a significant anti-hypertensive effect in this particular experiment.

IX.G) EFFECTS OF THE LYOPHILIZED OF THE HYDRO-ALCOHOLIC-ETHYL ACETATE EXTRACT OBTAINED FROM THE DECOCTION OF VITIS VINIFERA SKIN IN THE ARTERIAL HYPERTENSION INDUCED BY INHIBITION OF NITRIC OXIDE SYNTHESIS IN RATS

[0047] Adult, male Wistar rats (250-350 g, n=11) were kept cage with no more than 4 animals por cage. Mean arterial blood pressure was measured noninvasively, by Letica (Barcelona) equipment. A cuff and a sensor placed around the tail of the animal were connected to the equipment. The cuff is inflated automatically in order the measure the mean arterial blood pressure.

[0048] The animals were divided in two groups, were submitted to a period of adaptation of the experimental conditions, for measurement the mean arterial blood pressure. During the adaptation period, the animals received tap water and food “ad libitum”, and the daily intake of water was estimated. Body weight was estimated three times a week.

[0049] After the basal arterial pressure levels are obtained, both groups were treated orally with 70 mg/kg/day L-NAME in drinking water. After the arterial blood pressure reached high levels, due to L-NAME treatment, one group (n=5) received only L-NAME and the other group (n=6) was treated with L-NAME plus the extract, 100 mg/kg/day orally. This treatment induced a significant reduction of the high levels of arterial pressure, when compared with the control group, as showed in FIG. 7 .

PHARMACOTECHNICAL ASPECTS OF THE PREPARATION OF CAPSULES AND TABLETS CONTAINING THE DRY RESIDUE OF THE VARIOUS FRACTIONS OBTAINED FROM SKIN OF THE FRUITS OF VITIS LABRUSCA AND VITIS VINIFERA

[0050] The capsules and/or tablets containing 100 to 500 mg of lyophilized of Vitis labrusca or Vitis vinifera were obtained according to the usual pharmacotechnical procedures. The capsules were obtained in order to contain 100 to 500 mg, for instance 250 mg of the lyophilized plus cornstarch and colloidal silicon dioxide. Each capsule could have the following composition: 1

[0051] Cornstarch was added to complete the total mass of the capsule to approximately 450 mg. Colloidal silicon dioxide was used to adsorb humidity and to facilitate the preparation of the capsules.