RU2623872C1 - Means for prevention and treatment of type 2 diabetes, metabolic syndrome - Google Patents

Abstract

FIELD: pharmacology.

SUBSTANCE: means for prevention and treatment of type 2 diabetes, metabolic syndrome is dry red wine made from the fruits of red grape varieties and dry or fresh leaves of Gynura Procumbens, which is obtained by mixing crushed grapes and crushed Gynura Procumbens fresh leaves or Gynura Procumbens dry leaves, at the grapes:Gynura Procumbens fresh leaves ratio of (30-100):1 or grapes:Gynura Procumbens dry leaves ration of (200-500):1, fermentation, mash separation from pulp, subsequent fermentation, wine separation from sediment.

EFFECT: means has an increased effect in terms of prevention and treatment of type 2 diabetes, metabolic syndrome.

10 ex, 2 tbl

Description

The present invention relates to the pharmaceutical industry, and in particular to an agent for the prevention and treatment of type II diabetes, metabolic syndrome.

Metabolic syndrome is a group of risk factors for cardiovascular disease and diabetes. Disorders that are part of the metabolic syndrome, go unnoticed for a long time, often begin to form in childhood and adolescence, inevitably leading to atherosclerotic diseases, diabetes, arterial hypertension. Often, patients with obesity, a "slightly" increased glucose level, and blood pressure at the upper limit of the norm are not given proper attention. Only when these risk factors turn into a serious illness does the patient receive health attention.

It is important that risk factors are identified and corrected as early as possible before they lead to cardiovascular catastrophes.

For the convenience of patients and practitioners, clear criteria have been established that allow, with a minimal examination, to diagnose metabolic syndrome. Currently, most doctors use the single definition of the metabolic syndrome proposed by the International Diabetes Federation: a combination of abdominal obesity and any two additional criteria (dyslipidemia, carbohydrate metabolism disorders, hypertension).

“Metabolic Syndrome”, also called “Syndrome X”, “Insulin Resistance Syndrome” or “Deadly Quartet”, is characterized by summing up the risk factors for cardiovascular disease, stroke and / or type II diabetes mellitus. Metabolic syndrome may be due to overproduction of cortisol, a stress hormone that causes the accumulation of adipose tissue in the abdominal cavity and insulin resistance. Currently, patients with metabolic syndrome are not recommended drug therapy. The risk factors that characterize metabolic syndrome include an increased amount of adipose tissue inside the abdominal cavity (abdominal obesity), insulin resistance along with an increased risk of diabetes, hyperinsulinemia, high blood fats, high blood pressure and high serum lipids. In the National Adult Cholesterol Education and Treatment Program (ATP III), metabolic syndrome was defined as the presence of at least three of the following risk factors in patients:

However, the World Health Organization defines metabolic syndrome as the presence of diabetes / insulin resistance in individuals and at least two of the following risk factors: high ratio of waist circumference to hip circumference; high triglycerides or low HDL cholesterol; high blood pressure; and also a high rate of excretion of albumin in the urine.

Conditions related to the metabolic syndrome include type II diabetes mellitus, dyslipoproteinemia, myocardial infarction, stroke and other atherosclerotic diseases, as well as risk factors for these diseases, including insulin resistance in general, and abdominal obesity due to the accumulation of intraperitoneal fat, increased serum lipids and glucose levels, increased diastolic and / or systolic blood pressure and hypertension.

The treatment of patients with a condition preceding hypertension and patients with metabolic syndrome contributes to the adoption of a healthy lifestyle, which includes maintaining a healthy mass; physical activity; and following a healthy diet.

Diabetes is a systemic metabolic disorder that occurs under the influence of numerous environmental and genetic factors and is related to a condition characterized by abnormally high levels of glucose in the blood due to absolute or relative insulin deficiency in the body. Complications of diabetes include, for example, hypoglycemia, ketoacidosis, hyperosmolar coma, macrovascular complications, diabetic retinopathy, diabetic neuropathy, diabetic nephropathy, and the like.

The World Health Organization (WHO) estimates that more than 190 million people worldwide have diabetes, and this number is constantly growing. This disease can be divided into two main subclasses: insulin-dependent diabetes mellitus (IDDM), also known as type I diabetes, and non-insulin-dependent diabetes mellitus (NIDDM), also known as type II diabetes or adult diabetes.

In adult diabetes (type II), high blood sugar levels cause a recognizable increase in blood osmolality, which causes confusion and serious conditions such as unconsciousness. Long-term complications are observed in both small and large blood vessels, which affects the heart, kidneys and eyes, and also affects the sensory nerves (especially in the legs). Damage also occurs in the autonomic nervous system and is expressed in impotence, infertility and disorders of the digestive system, heart and blood vessels.

A significant proportion of adults with diabetes also suffer from high blood pressure. Both diseases need to be given attention, because both of these accelerate degeneration with complications, especially in the blood vessels and heart.

Another condition associated with diabetes as a secondary cause is hypertriglyceridemia, a common lipid disorder that is often associated with additional lipid and metabolic disorders.

Regulation of diet and exercise and / or treatment with insulin or hypoglycemic drugs has been used to control both diabetes and triglycerides. Treatment with these substances is successful in some cases, but the mortality rate continues to rise. Treatment with insulin and hypoglycemic agents (such as sulfonylurea, biguanides and α-glucosidase inhibitors) provides symptom relief rather than diabetes, and is associated with side effects. Side effects of sulfonylurea include hypoglycemia, kidney and liver disease, gastrointestinal disease, increased mortality from cardiovascular disorders, dizziness, drowsiness, and others. The main side effects of biguanides are lactic acidosis and increased mortality from cardiovascular disorders. Side effects of α-glucosidase inhibitors include gastrointestinal side effects and hypoglycemia.

In addition to traditional treatments based on insulin injections or oral drug therapy, natural products, including plant materials, have been tested in alternative treatments for conditions such as diabetes.

Thus, it is known that the branches and bark of Mulberry alba (white mulberry) are used to lower blood pressure (Enkhmaa et al., J. Nutr. 2005, vol. 135, No. 4, pp. 729-734). Leaves of M. indica L. have been shown to have hypoglycemic, hypotensive, and diuretic properties, although there was no apparent effect on the concentration of glycosylated hemoglobin (HbA (1) c) in patients with diabetes (Andallu et al., Clin Chim Acta. 2001 Dec ; 314 (1-2): 47-53).

Canella winteriana (commonly known as cinnamon, cinnamon bark, cinnamon bark, pepper cinnamon, wild cinnamon) and Cinnnamomum cassia (cinnamon tree) are thought to be good for maintaining normal blood sugar levels as well as cholesterol levels (Aaron W Jensen, “Cinnamon Reduces Blood Sugar and Cholesterol Levels”, InsuLifeTM). However, despite the fact that daily doses of cinnamon cause a slight decrease in blood sugar, total cholesterol, triglycerides and even lower levels of low density lipoproteins, an increased dose of cinnamon does not effectively reduce the various blood levels described above compared to lower doses of cinnamon (Alam et al., Diabetes Care 26: 3215-3218, 2003).

Artemisia is a large, diverse plant genus with 200-400 species belonging to the Asteraceae family, which grows in a temperate climate, usually in dry or semi-dry natural habitats. In US Pat. No. 6,350,478 B1, 02/26/2002, extracts of Artemisia judaica and their fractions are disclosed, and some fractions have been found to be insulinomimetics, while others have glucagon antagonist properties.

Another study on the effectiveness of dietary supplements with herbal medicines on carbohydrate metabolism in humans (Cefalu et al., Endocr. Metab Immune Disord Drug Targets, 2008, 8 (2): 78-81) evaluated the effectiveness of bitter melon (Momordica charantia ), fenugreek (Trigonella foenum graecum), Gymnema sylvestre, Indian coccinia (Coccinia indica), Mexican cactus or prickly pear (Opuntia streptacantha), ginseng, aloe vera, Russian tarragon (Artemisia dracunculus) and researchers () to the conclusion that there is insufficient evidence based on clinical studies REPRESENTATIONS for any considered herbal medicines and it is premature to urge the use of a particular plant in the treatment of glucose or other risk factors.

Thus, there is a continuing need for the discovery of natural medicines for diabetes and / or hypertriglyceridemia, which are highly effective in lowering blood pressure, lowering blood glucose and / or triglycerides.

The authors of the present invention developed a composition obtained from the leaves of Gynura Procumbens and fruits of red grape varieties, which is effective for the prevention and treatment of type II diabetes, metabolic syndrome. The proposed composition can also be used in the food industry.

Gynura Procumbens from the family of asters was born many centuries ago on the African continent, then the plant successfully migrated to Asia, where it was appreciated by the healers of China and Southeast Asia. It was used simply as food and at the same time as a cure for disease. In Southeast Asia, Ginura is part of a balanced diet and healthy lifestyle. Phytochemical studies confirm that the extract from the leaves of Ginura (ginura) is an excellent natural source of many potent biologically active components, including flavonoids, carotenoids, alkaloids and essential oils. An active effect on metabolic processes puts Ginuru in a number of other medicinal plants, which are actively used as primary protection for cancer patients before chemotherapy and radiotherapy.

The results of clinical studies confirm the therapeutic effect of Ginura in diabetes, kidney disease, gastroenterological and cardiovascular diseases. The plant has pronounced antioxidant properties, inhibits the growth of cancer cells in the body, has an anti-inflammatory effect and helps to increase reproductive function.

In TAN HL et al. Gynura procumbens: An Overview of the Biological Activities // Front Pharmacol. 2016, Mar 15, presents a report on a study of Gynura Procumbens and tools based on it. This document discloses that Gynura Procumbens has great therapeutic potential, a variety of pharmacological activities: antihypertensive, cardioprotective, antihyperglycemic, anti-cancer, antibacterial, antioxidant, general protective, anti-inflammatory. Gynura Procumbens is characterized by a high content of biologically active substances: flavonoids, glycosides, etc. This document describes the use of the aqueous and alcoholic extract of Gynura Procumbens leaves. The disadvantage of the funds from Gynura Procumbens described in this document is that the funds do not effectively lower the level of cholesterol, glucose in the blood. The result is insufficient clinical results in the prevention and treatment of type 2 diabetes, as well as metabolic syndrome.

It is also known from various sources of information that the consumption of red wine in moderation in diabetics significantly reduces oxidative stress and improves cardiac function. Moderate consumption of wine can have a beneficial effect in preventing cardiovascular diseases after myocardial infarction (see, for example, patents CN 103122295 A, 05/29/2013, CN 104673573 A, 06/03/2015 and article MARFELLA R et al. Effect of moderate red wine intake on cardiac prognosis after recent acute myocarction of subjects with Type 2 diabetes mellitus // Diabet Med. 2006 Sep. 23 (9): 974-81).

Resveratrol is also known to be useful in treating diabetes and its complications. Resveratrol is found in sufficient quantities in red wine and red grapes (article VALLIANOU NG et. Al. Resveratrol and diabetes // Rev Diabet Stud. 2013 Winter, 10 (4): 236-42).

The objective of the proposed invention was to develop a tool based on the plant Gynura Procumbens and fruits of red grape varieties, the use of which will lead to high clinical results in the treatment and prevention of type 2 diabetes and metabolic syndrome.

The technical result is that the proposed tool, which is a dry wine obtained from fresh or dry leaves of Gynura Procumbens and fruits of red grape varieties, has increased effectiveness in the prevention and treatment of type 2 diabetes and metabolic syndrome.

The technical result is achieved by the fact that the proposed tool for the prevention and treatment of type 2 diabetes, as well as metabolic syndrome, is a dry wine obtained from fruits of red grape varieties and fresh leaves of Gynura Procumbens or dried leaves of Gynura Procumbens, soaked previously in water, by mixing the fragmented grape fruits and crushed leaves of Gynura Procumbens in the ratio grape fruits: fresh leaves of Gynura Procumbens (30-100): 1 or grapes: dry leaves of Gynura Procumbens (200-500): 1, fermentation, wort separation from pulp, rice further fermentation, separation of wine from sediment. To enhance the fermentation process in the dry leaves of Gynura Procumbens, they are pre-soaked with water before mixing with grape fruits.

A unique feature of the proposed tool for the prevention and treatment of type 2 diabetes, metabolic syndrome is the use of a medicinal plant - Gynura Procumbens leaves in winemaking and the extraction of bioactive plant ingredients by co-fermentation with red grape fruits.

The method of obtaining funds

Example 1

Dry wine is obtained using traditional techniques for obtaining dry wines from red grape varieties.

They use grape varieties such as Cabernet Sauvignon, Saperavi, Odessa Black, Rubin Tairovsky, Sereksia, Bastrado, Magarachsky, Mattress, Limberger, red sable varieties, red varieties of hybrids from direct producers - Zeibels of various numbers, Kuderk and others.

The fruits of the grapes are crushed. Fresh or dry leaves of Gynura Procumbens are crushed to 3-8 mm.

For 1000 kg of grape fruit, 33.3 kg of fresh leaves or 5 kg of dry leaves of Gynura Procumbens are used.

To start fermentation in the dry leaves of Gynura Procumbens, they are pre-soaked in water. To do this, the crushed dried leaves are placed in a container and 5-30 liters of water are poured at room temperature, mixed to a homogeneous mass and infused for about one hour.

Next, crushed fresh or soaked leaves of Gynura Procumbens are divided into three equal parts and introduced into the fermentation tank as follows:

1. The first part - after filling the tank with crushed grapes by 25%;

2. The second part - after filling the tank by 50%;

3. The third part - after filling the tank by 75%.

Such loading is advantageous, as it allows more evenly distributed plant components for their further fermentation, but is not limited to this. To obtain dry wine from the fruits of red grape varieties and fresh or dry leaves of Gynura Procumbens, the components can also be mixed in any order.

The next day, fermentation usually begins. During fermentation, it is necessary to stir and immerse sticky particles of the pulp - “caps” as often as possible, which is done at least three times a day. Stirring is carried out until the entire "cap" is broken into small pieces and evenly distributed throughout the volume of the entire liquid.

To control the process, the temperature of the fermented mass is measured daily in the morning and evening. Measurement is carried out in the center of the tank at the level of the "cap" and liquid. It is recommended not to allow temperature levels above 30 ° C.

Separation of the wort from the pulp (a mixture of plant material: seeds and skin of grapes, leaves of Gynura Procumbens) occurs after the end of the primary fermentation, which lasts 7-14 days.

In the wort separation, the resulting young wine is poured into containers to complete the fermentation, which usually lasts 1.5-2.5 months. During this period, the wine parameters are monitored daily using well-known technological methods.

At the end of the fermentation, the wine is separated from the precipitate and poured into oak barrels, followed by aging for 10-12 months.

After aging in oak barrels, the wine is bottled.

In the fermentation process, the bioactive components are extracted from the leaves of Gynura Procumbens into wine, which determines the claimed therapeutic effects of the final product.

Example 2

The method of obtaining funds is carried out in the same way as in example 1, and differs in that 25 kg of fresh leaves of Gynura Procumbens or 2.5 kg of dry leaves of Gynura Procumbens are used per 1000 kg of grape fruit.

Example 3

The method of obtaining funds is carried out in the same way as in example 1, and differs in that per 1000 kg of grape fruit use 10 kg of fresh leaves of Gynura Procumbens or 2 kg of dry leaves of Gynura Procumbens.

Example 4

The method of obtaining funds is carried out in the same way as in example 1, and differs in that per 1000 kg of grape fruit use 35.7 kg of fresh leaves of Gynura Procumbens or 5.5 kg of dry leaves of Gynura Procumbens.

Example 5

The method of obtaining funds is carried out in the same way as in example 1, and is characterized in that 9 kg of fresh Gynura Procumbens leaves or 1.85 kg of dry Gynura Procumbens leaves are used per 1000 kg of grape fruit.

Obtained in examples 1-5, the agents were investigated for the content of biologically active substances by the method of spectral-dynamic analysis and mass spectrometry.

In clinical studies, it was found that the optimal ratio of red grape fruit to fresh or dry leaves of Gynura Procumbens is (30-100): 1 or (200-500): 1, respectively.

When using other ratios of components to obtain the proposed tool, the content of biologically active substances in the final product was reduced.

The funds obtained in examples 1-3, unlike standard dry wines, are saturated with phytosterols, which are a key factor in lowering cholesterol, as well as transferring a complex of biological substances through cell membranes; succinic acid, omega-3 fatty acids, which improve cell function by restoring membrane function; bioflavonoids; amino acids such as tryptophan, phenylalanine, aspartic acid.

Clinical trials

The proposed tool obtained in examples 1-3, is prescribed in 50-60 ml twice a day 30 minutes before meals for 21 days.

Example 6. The effect of the funds obtained in examples 1-3, on blood pressure in patients with metabolic syndrome

The effects of the proposed agent used according to the present invention on blood pressure were studied in 32 patients with a diagnosis of metabolic syndrome. The study included an equal number of men and women aged 20 to 50 years. Metabolic syndrome was diagnosed based on criteria defined by the ATP III National Adult Cholesterol Education and Treatment Program. At least three of the following symptoms were observed in each patient: 1) fasting blood sugar> 110 mg / dl, 2) HDL <40 mg / dl in men and <45 mg / dl in women, 3) blood pressure> 130/85 and 4) abdominal obesity> 102 cm for men and> 88 cm for women. Individuals were excluded if they were current or former smokers (<3 years); took anti-inflammatory or antihypertensive drugs or consumed over-the-counter antioxidant compounds.

Patients were randomly combined into four groups of eight people, and depending on the destination for their group, they received one of the following funds.

Group 1 received the proposed tool in 50-60 ml twice a day 30 minutes before meals.

Group 2 received 50-60 ml twice daily aqueous extract of Gynura Procumbens leaves 30 minutes before meals.

Group 3 received a placebo twice daily 30 minutes before meals.

Patients received a sufficient amount of funds, taking the same dose once a day for twenty-one days. At the end of this period, blood pressure was measured. Ambulatory blood pressure values were recorded over a period of 12 hours at the start of the study and again after three weeks. The method was non-invasive and included placing a cuff on the shoulder to measure blood pressure. The cuff was paired with an automatic, FDA-approved inflator worn on the belt.

Table 1 presents blood pressure data for three groups of individuals with metabolic syndrome. In patients receiving the proposed tool used according to the present invention, a significant decrease in systolic and diastolic pressure was observed. In the group that received the aqueous extract of Gynura Procumbens leaves, a slight decrease in pressure was observed, in the placebo group, there were no significant changes.

Since the diagnosis of metabolic syndrome is based on the presence of three of the above risk factors (one of which is blood pressure), the individuals in the study were not randomly divided in terms of blood pressure. As such, the mean arterial pressure in the three groups was not similar (but varied within a narrow range).

This study showed that the proposed tool effectively reduces both systolic and diastolic blood pressure in patients with metabolic syndrome. The decrease in blood pressure is statistically significant for the funds obtained in examples 1-3. Indeed, changes in blood pressure observed when using the funds obtained in examples 1-3 were comparable to those observed in large clinical trials using pharmaceuticals.

Example 7. The effect of the funds obtained in examples 1-3, on the level of oxidized LDL in individuals with metabolic syndrome

The effects of the agents used according to the present invention on the level of oxidized LDL were studied in the same 32 patients diagnosed with metabolic syndrome, as described above. The concentration of oxidized LDL was measured at the beginning of the study and again three weeks after treatment. To measure the concentration of oxidized LDL, a blood sample was taken from each individual and analyzed.

The studies showed a slight decrease in the level of oxidized LDL in the case of placebo and patients taking an aqueous extract of Gynura Procumbens leaves, a statistically significant decrease (p <0.05) in the level of oxidized LDL in individuals who received the funds obtained in examples 1-3 used according to the present invention. A significant decrease in the concentration of oxidized LDL was shown in patients, primarily for those who initially had higher levels of oxidized LDL.

This study showed that the proposed tool effectively reduces the concentration of oxidized LDL in plasma in individuals with metabolic syndrome.

Example 8. The effect of funds according to examples 1-3 on patients with a condition prior to hypertension

The effects of the agent used according to the present invention were studied in 28 patients with a diagnosis of a condition prior to hypertension. The study included an equal number of men and women aged 30 to 60 years. The condition prior to hypertension was diagnosed based on the seventh report of the Joint National Committee for the Prevention, Detection, Assessment and Treatment of High Blood Pressure. Each patient had a systolic pressure of 120 to 139 mm Hg. and / or diastolic pressure from 81 to 89 mm Hg Patients were excluded if they were current or former smokers (<3 years); took anti-inflammatory or antihypertensive drugs or consumed over-the-counter antioxidant compounds.

Patients were randomly combined in terms of gender in two groups of fourteen people, and depending on the destination for their group, they received one of the following means.

Group 1 - the product obtained in examples 1-3, 50-60 ml 2 times a day 30 minutes before meals.

Group 2 received an aqueous extract of Gynura Procumbens leaves 50-60 ml 2 times a day 30 minutes before meals.

Patients received a sufficient amount of funds, taking the same dose once a day for the next eight weeks. At the end of this period, blood pressure was measured. Ambulatory blood pressure values were recorded over a period of 12 hours at the start of the study and again after eight weeks. The method was non-invasive and included placing a cuff on the shoulder to measure blood pressure. The cuff was paired with an automatic, FDA-approved inflator worn on the belt.

Table 2 presents blood pressure data for two groups of patients with a condition prior to hypertension. The initial values of blood pressure for the two groups did not significantly differ. In patients who daily received the proposed tool used according to the present invention, a significant decrease in both systolic and diastolic pressure was observed; whereas in the group receiving the aqueous extract of Gynura Procumbens leaves, there were no significant changes. Summarized data is given below. Values are presented in mmHg. (mean ± SEM).

This study showed that the proposed tool reduces systolic and diastolic blood pressure in patients with a condition prior to hypertension. A decrease in blood pressure is statistically significant. Indeed, changes in blood pressure observed when using the proposed tool were comparable to those observed in large clinical trials using pharmaceuticals.

Example 9. The treatment of type 2 diabetes using the funds obtained in example 1

Patient K.E. Born in 1954 with a 23-year history of type 2 diabetes (started as gestational diabetes), complicated by proliferative retinopathy (1955), cataract of the left eye (operated on) and later by vinrectomy of the right, periodically increasing urinary albumin excretion rate, peripheral polyneuropathy. Vascular insufficiency, thibial vein thrombectomy, proximal anastomotic jump to the posterior tibial artery. Rheumatic polyarthritis. Diabetes has been treated with subcutaneous insulin for the past ten years. Drug therapy included insulin, convertin 5 mg daily, coumadin 2.5 mg daily, and lipitor (atrovastatin) 10 mg daily. To the treatment was added the use of the proposed drug in 50-60 ml 2 times a day 30 minutes before meals for 15 days. Subsequently, insulin administration was gradually reduced. After 21 days of taking the proposed drug, the blood glucose level significantly decreased from 180 ± 20 to 115 ± 20 mg%, HbA1C decreased from 10.6 to 6.27, and the indicators constantly improved.

Example 10. Treatment of a patient with diabetes of a patient of type 2 with the agent obtained in example 3.

Patient T.A. Born in 1951, height 168 cm; weight is 69 kg. BMI 24.3 Presented with a history of treating type 2 diabetes for 6 years, hypertension, hyperuricemia and dyslipidemia, blood glucose 350 ± 50, HBA1c 10.69, total cholesterol 262 mg / dl and triglyceride 316 mg / dl, uric acid - 4.6 mg / dl. Diabetes was treated with glyburide and pranase (acarbose) with the appointment of three times a day each, including a strict diet and exercise.

The patient was prescribed treatment with the proposed drug in 50-60 ml 2 times a day 30 minutes before meals for 21 days. Constantly monitored the level of glucose in the blood. After 19 days of taking the proposed drug, the glucose level in the blood significantly decreased from 230 to 137 mg%, HbA1C decreased from 10.7 to 8.5.

After the treatment, fasting blood glucose was constant 100 ± 10 mg / dl.

Twenty-six patients with a diagnosis of type 2 diabetes were examined. After the treatment using the proposed funds obtained in examples 1-3, the fasting blood glucose level dropped to 44.5 ± 10% after 21 days of treatment, HbA1C levels dropped to 33.96 ± 10% after 21 days of treatment.

When using the funds obtained in examples 4,5, there is a slight decrease in blood glucose and HbA1C.

Using the proposed tool has shown high efficiency in normalizing blood glucose levels, reducing HbA1C, normalizing lipid metabolism, blood pressure.

Thus, the proposed tool is effective for the prevention and treatment of type 2 diabetes mellitus, metabolic syndrome.

Claims (1)

An agent for the prevention and treatment of type 2 diabetes, metabolic syndrome, which is a dry wine made from red grapes and dry or fresh leaves of Gynura Procumbens, obtained by mixing crushed grapes and crushed fresh leaves of Gynura Procumbens or dried leaves of Gynura Procumbens, soaked previously in water, in the ratio grape fruit: fresh leaves of Gynura Procumbens (30-100): 1 or grape fruit: dry leaves of Gynura Procumbens (200-500): 1, fermentation, separation of wort from pulp, subsequent fermentation, separation of wine from sieges a.