FR2465484A1 - Extract of e.g. eugenia jambolana lamarck - has antidiabetic, hypoglycaemic, hypolipaemic, weight reducing anorexic and anti-cataract activity - Google Patents

Abstract

Plant extract, containing a sterolic and polyphenolic complex mixture, prepared from the seeds or bark C from branches, trink and roots) of Eugenia Jambolana Lamarck or related varieties of Eugenia or from the internal bark (cambium) of Anacardium Occidentale L is new. The extract is prepared by treatment of the finely ground material with heated water so as to remove unwanted water-soluble fractions, filtration of the obtained suspension, extn. of the water cake with an organic solvent (pref. an alkanol, esp. ethanol) at first in the cold and then with heating; filtration of the remaining solids and evaporation to dryness of the filtrate to obtain the sterolic and polyphenolic complex mixt. The extract has antidiabetic, hypoglycemic, hypolipaemic, weight reducing, anoersic and anti-cataract activity. It may be administered oralloy in unit doses of 0.5-3 g. Toxicity, pharmacological and clinical tests are described.

Description

 The subject of the present invention is a medicament active on carbohydrate and lipid metabolism, resulting in particular in an antidiabetic and lipid-lowering activity and also having an activity against cataracts, containing as active principle, a new mixed polyphenolic and sterolic complex extracted from seeds and barks of Eugenia Jambolana Lamarck and other similar varieties of Eugenia, as well as the inner bark (cambium) of Anacardium Occidentale L.

Eugenia Jambolana Lamarck is a tree belonging to the myrtaceae family. It is also known by the following names: Eugenia Jambolanum, Myrtus
Cuminis Linné, Syzygium Jambolanum de Candolle, Eugenia
Cumini Bruce, Syzygium Cumini Skeels, Jambolan.

In Madagascar, its vernacular name is Rotra or Rotravazaha.

The other varieties of Eugenia which can be used are: Eugenia Parkeri Bak., Eugenia Hovarum H. Perrier,
Eugenia Condensata Bak., Eugenia Emirnensis Bak.,
Eugenia Coursiana H. Perrier, Eugenia Danguyana H.

Perrier variety Rotranala H. Perrier, Eugenia Sakalavorum H. Perrier.

Anacardium Occidentale L. is a tree belonging to the anacardiaceae family, a tribe of mangiferae.

 In Madagascar, its vernacular name is Mahabibo.

 Empirical knowledge of the use of crushed or infused raw seeds by Eugenia Jambolana Lamarck in the treatment of symptoms related to diabetes, in particular polyuria and polydipsia, is old, but no active product devoid of toxicity has never been been marketed.

Authors who have attempted to show the hypoglycemic or antidiabetic power of different organs of Eugenia Jambolana Lamarck have never described the method of preparation and the characteristics of the active ingredients they used.

The present invention aims to provide a new drug based on a mixed polyphenolic and sterolic complex containing all the antidiabetic activity but free of the toxic molecules of crude extract, responsible for intestinal transit disorders and inflammatory manifestations in the tube digestive and transient hyperglycemia.

This new drug has important antidiabetic properties resulting in the fRis of its hypoglycemic, lipid-lowering, slimming and appetite suppressant properties and an activity against cataracts.

The medicament according to the invention is remarkable in particular in that it contains, as active principle, a mixed polyphenolic and sterolic complex extracted either from seeds and bark originating from the branches, trunk and roots of Eugenia Jambolana
Lamarck and the other varieties of Eugenia listed above, namely the inner bark (cambium) of Anacardium
Occidental L.

The plant extract according to the invention is characterized in that it contains a mixed polyphenolic and sterolic complex prepared from seeds or bark of Eugenia Jambolana Lamarck or from other varieties close to Eugenia, or from inner bark (cambium) of
Anarcadium Occidentale L, by treatment with 1V water, under heating of the ground starting material, in order to eliminate the undesirable water-soluble fractions, filtration of the suspension obtained, then extraction of the filter cake using an all organic solvent first cold then under heating, filtration of the insoluble material and evaporation to dryness of the organic solvent to obtain the extract containing the mixed polyphenolic and sterolic complex.

The extraction of molecules acting synergistically only in the treatment of diabetes is carried out in two successive essential stages: on the one hand, elimination by extraction with water and under heating of molecules with a strong polar character thanks to their water solubility, on the other hand part, recovery of the remaining insoluble part by means of an organic solvent allowing a large grouping of molecules of different polarity, in particular a lower alkanol such as ethanol.

 The preparation of the active principle of the medicament of the invention is carried out according to the protocol according to the various organs of the plant, seeds and bark are dehydrated to 95% by passage of hot pulsed air (maximum temperature 600C) and kept in airtight containers. protected from light, they are ground before preparation and sieved to 160 (according to standard NF x 11 - 501).

The first extraction is carried out with distilled water. 100 kg of powder are extracted by 300 liters of water brought to 80 ° C. in thirty minutes. This temperature being reached, without stopping stirring, the suspension is cooled to the temperature from 20 to 250C (maximum). One then proceeds to a rapid filtration followed by a moderate pressing of the grounds. Qn then proceeds to extract the molecules constituting the anti-diabetic extract. The filter cake from the previous treatment, weighing 110 to 115 kg, is macerated for 48 hours at room temperature in 200 liters of ethyl alcohol at 950 GL. The alcohol is heated at the end of maceration, then kept at boiling point 60 minutes. Hot filtration is then carried out followed by pressing of the insoluble material. The solution obtained is evaporated to dryness under reduced pressure at a temperature below 550C to obtain the desired extract.

 The yield of obtaining the dry extract is 8 to 9.5 <with respect to the dry and ground starting material and constitutes the active fraction in the treatment of diabetes.

The active principle of the drug is in the form of a powder of brown-red color, astringent flavor, slightly aromatic odor. The solubility of the active ingredient at 200 C is 63 to 70% in liteau, 36 to 42% in alcohol, less than 0.1% in chloroform. The pH of 1 g of extract diluted in 20 ml of distilled water is between 3.5 and 4.5.

CHROMATOGRAPHIC STUDY OF THE DIABETIC COMPLEX
The complex developed consists of a set of molecules acting in synergy and consisting essentially of bodies belonging mainly to the groups of sterols and polyphenols.

 1 - The total complex is chromatographed on a thin layer of polyamide using ternary eluents such as butanol / acetic acid / water.

 In the 30/55/15 system, phenolic compounds are eluted mainly: aglycones and heterosides. The complex is thus identified by molecules which can simultaneously be revealed by means of several characterization reagents such as the acid blue salt B in OH medium, Mn04K in OH medium, Cl2Fe. Their Rf are respectively 0.22 - 0.35 - 0.54 - 0.88 - 0.96.

The hydrolysis of the isolated polar fraction, carried out using HCl, gives the Rf of 0.22 - 0.35 - 0.54 - 0.73.

Rf 0.54 is used as a reference and is identified with gallic acid.

 2 - The molecules used in the composition of the complex being of different polarities, the hexane extraction of the sterols is carried out, before chromatographic separation on a thin layer of silica G.

 The orthophosphoric and phosphomolybdic reagents, the para-toluene sulfonic reagent, the TTC, the modified Millon reagent, the benzoyl chloride allow the characterization of 5 main molecules having for Rf: 0.37 - 0.39 - 0.53 - 0.86.

Sitosterol of Rf 0.55 is considered as the reference molecule.

 SPECTROMETRY U.V.

max. = 277.5 mA E0.10 g / 1 = 1.012- (methanol)
1 cm
ailments. = 225 m E0.10 g / l = gel (methanol)
- 0.801
1 cm
The medicament of the invention is usually presented for oral administration in the form of unit doses of antidiabetic extract powder, each dose being bagged in a sealed and sealed sachet.

 Due to its very low toxicity, the daily doses administered may vary according to the clinical cases treated from 1 g to 6 g of antidiabetic extract per day.

 The unit doses of antidiabetic extract powder are presented in the form of individual sachets containing from 0.5 to 3 g and in particular 1 or 2 g of extract.

Other pharmaceutical forms of the drug such as tablets, coated tablets, cachets, capsules, oral solution, can be produced. The preparation of these formulations is carried out in a conventional manner for the manufacture of these pharmaceutical forms.

The active principle of the medicament of the invention has remarkable anti-diabetic, lipid-lowering and overweight-reducing properties, anorectic and cataract properties demonstrated by the results of the pharmacological and clinical trials given below after the results of the toxicological tests.

A - TOXICOLOGICAL STUDY
This study focused on acute toxicity, chronic toxicity, delayed toxicity, general tolerance.

Tests carried out on mice, rats and rabbits have shown that the medicament of the invention administered both by the oral and intraperitoneal routes has no toxic effect and that it is perfectly tolerated.

The LD50 for the oral route in mice is, for information, 5.5 g / kg of body weight.

B - PHARMACOLOGICAL STUDY
1 - The hvDiglycemic action
It has been shown by the action of the anti-diabetic extract on the oral hyperglycemia test caused by normal rabbits.

The rabbit is put on glueidic overload with a dose of 1 g / kg of body weight of glucose and simultaneously receives the antidiabetic extract. Blood sugar levels are measured every thirty minutes for two hours.

Under these experimental conditions, there is a 20% drop in blood sugar at times 60, 90 and 120 minutes of the test.

2 - Absence of action on the blood sugar base of the normal animal
The anti-diabetic extract administered for a prolonged period in normal rats does not change the basic blood sugar.

3 - Anti-diabetic action
It has been demonstrated in the case of experimental diabetes and in the case of spontaneous diabetes. Experimental diabetes was determined by alloxan in rats and mice.

The drug of the invention administered daily to rats or to mice made diabetic by alloxan determines a significant drop in blood sugar level which, even according to the doses used, can bring blood sugar back to normal figures after two months of treatment.

There is also a disappearance of glycosuria and a decrease in mortality.

 Spontaneous diabetes is that of the psammomys obesus or desert rat and that of the obese and diabetic mouse (db +).

In the rodent psammomys obesus of the sub-desert regions becoming diabetic as soon as it is withdrawn from its natural biotope, the administration of the antidiabetic extract as a preventive measure prevents the appearance of signs of diabetes; blood sugar remains normal, glycosuria does not appear. Given as a curative after two months of evolution of diabetes, it determines a return of blood sugar to around normal with disappearance of glycosuria. In both cases, it allows normal longevity and reproduction of animals.

 In obese and diabetic mice (db +), the drug of the invention given for almost two months significantly lowers blood sugar and eliminates glycosuria.

4 - Effects on polyuria, polydipsia and polyphagia
In diabetic animals (experimental or spontaneous diabetes), a decrease in polydipsia, polyuria and polyphagia was observed during all of the experiments.

5 - Anorectic effect
The anti-diabetic extract exerts in the normal rat an important appetite suppressant equivalent to that of D-amphetamine tartrate when it is administered intraperitoneally. By mouth, the appetite suppressant can only be observed in high doses.

6 - Action on the psammomys cataract (desert rat)
Preventive administration or after two months of development of diabetes, the antidiabetic extract, allows to note the absence of appearance or a progressive regression of the cataract of psammomys obesus. I1 does not seem that other anti-diabetic therapeutics exert this activity on the cataract which thus appears completely original.

7 - Action on weight reduction
The anti-diabetic extract determines a reduction in weight both in the obese and diabetic mice and in its normal heterozygous control. In both cases there is a decrease in food intake.

8 - Search for side effects
It has not been shown to have any side effect from the product, in particular on intestinal transit, the cardiovascular system and blood pressure, diuresis and elimination of electrolytes from normal animals.

The results of these tests show that the medicament of the invention has - has remarkable antidiabetic properties.
without exerting a hypoglycemic effect on blood sugar
of the normal subject, b - is capable of correcting hyperlipemia,
reduce overweight and exercise
anorectic effect, c - exercises a completely original activity of regres
Or prevention of cataracts.

The previous study highlighted the pharmacological properties of the medicament of the invention which, devoid of secondary actions which reduce its effectiveness, can be advantageously used in human therapy, as demonstrated by the following clinical study.

C - CLINICAL STUDY
Administered to diabetic patients, the medicament of the invention has shown remarkable efficiency. Its therapeutic action was judged on the regression of the clinical signs of diabetes and the improvement of the biological constants.

 Administered to patients with hyperlipemia or overweight, the efficacy was also very good.

Observation 1
Mme RAS ..., 49 years old, has a plethoric diabetes which has never been treated, with polydypsia, polyuria, polyphagia, asthenia, drowsiness, obesity: 71 kg for a height of 1 m 54. Blood sugar 2.23 g / l, glycosuria 32 g / l, cholesterolemia 3.07 g / l.

After 30 weeks of treatment with 3 sachets of 1 g of active principle per day, associated with a moderate hypoglucidique diet, we are witnessing a regression of clinical signs. Glycemia 0.96 g / l, negative glycosuria, cholesterolemia 2.07 g / l, weight 63 kg (loss of 8 kg).

Excellent tolerance.

Observation # 2
Mr RAM ..., 65, diabetic for 11 years, has been treated several times with insulin and various oral antidiabatic agents. Treatment has been discontinued for six months.

 There is an aggravation of the symptoms of diabetes, in particular of asthenia and especially there is an incomplete bilateral diabetic cataract. Glycemia 1.92 g / l, glycosuria 20 g / l, presence of ketones in the urine, cholesterolemia 3.10 g / l.

After 30 weeks of treatment with 3 sachets of 1 g per day associated with a moderate carbohydrate restriction, there is improvement in clinical signs and general condition. Glycemia 0.93 g / l, negative glycosuria, disappearance of ketonuria, cholesterolemia 2.18 g / l, but what is particularly remarkable is the regression of the cataract noted by the improvement of the visual condition of the patient and eye exams. Excellent tolerance.

Observation # 3
Mme RAS ....., 53 years old, has a plethora of diabetes, discovered 5 months ago, never treated4. She complains of asthenia, polydypsia, polyuria, polyphagia; there is obesity, o6 kg for a height of 1 m 51. Glycemia 1.70 g / l, glycosuria 15 g / l, cholesterolemia 3.27 g / l.

 After 13 weeks of treatment with 3 sachets of 1 g per day, without an associated diet (the diet is refused by the patient), we are witnessing a decline in clinical signs, in particular of asthenia. Glycemia 0.97 g / l, negative glycosuria, cholesterolemia 2.42 gXl, weight 61 kg (loss of 5 kg). Excellent tolerance.

Observation # 4
Mr. Ans ..., 52, has been diabetic for 2 1/2 years. He was treated with an oral anti-diabetic drug which has been discontinued for a year. There is an aggravation of the symp temples. The patient complains of great asthenia with sexual impotence. Glycemia 1.60 g / l, glycosuria 26 g / l, presence of ketones in the urine.

After 9 weeks of treatment with 3 sachets of 1 g per day, associated with a moderate carbohydrate restriction, there is one. improvement of the general state with resumption of a certain sexual activity. Blood sugar 0.98 g / l, negative glycosuria, negative ketonuria.

Excellent tolerance.

Observation # 5
Mme RA ...., 40, has had diabetes for 6 years treated with insulin for a few months, then with oral anti-diabetics for a year. The patient complains of polyuria, polydypsia, asthenia. Glycemia 1.46 g / l, glycosuria 14 g / l, cholesterolemia 4.32 g / l.

After 10 weeks of treatment with 3 sachets of 1 g per day associated with a moderate hypoglucidic diet, we are witnessing a regression of clinical signs, in particular of asthenia. Glycemia 0.99 g / l, negative glycosuria, cholesterolemia 2.70 g / l. Excellent tolerance.

Observation # 6
Mr. RALI ..., 40 years old, has had diabetes for 3 years stabilized by NN dimethyl biguanide. The blood sugar is 1 g / l. The drug of the invention is substituted for NN dimethyl biguanide at a dose of 2 sachets of 1 g per day. After 7 weeks of treatment, the blood sugar has not changed, it is 0.98 g / l. Excellent tolerance.

Observation n
Mr. RA ...., 56, has had diabetes for 8 years treated with insulin and then with chlorpropamide which stabilizes glycosuria at 1.12 g / l. The drug of the invention is replaced with chlorpropamide at a dose of 3 sachets of 1 g per day. After 4 weeks of treatment, the blood sugar has not changed, it is 1.05 g / l.

Excellent tolerance.

Observation # 8
Mrs. TAR ..., 21, has insulin-dependent diabetes treated with insulin for many years at a dose of 45 units per day. The gradual addition of the medicament of the invention to insulin at the rate of 3 sachets of 1 g per day makes it possible to reduce the dose of insulin to 25 units. Excellent tolerance.

Observation n
Mrs MAN ..., 57 years old, presents a significant overweight with hyperlipemia and hyperphagia. Its weight is 87 kg for a size of 1.59 m. Cholesterolemia 3.40 g / l, triglyceridemia 1.96 g / l. After 14 weeks of treatment at the rate of 3 sachets of 1 g per day, there is a reduction in weight of 11 kg, a decrease in cholesterolemia 2.61 g / l and triglyceridemia 1.27 g / l. Excellent tolerance. The patient notes an anorectic effect of the product.

Observation # 10
Mme RAVE ..., 49 years old, is overweight with hyperlipemia and hyperphagia. Its weight is 74 kg for a size of 1 m 54. Cholesterolemia 3> 15 g / l. Triglyceridemia 1.76 g / l. After 10 weeks of treatment at the rate of 3 sachets of 1 g per day, there is a reduction in weight of 7 kg, a fall in cholesterolemia 2.54 g / l and in triglyceridemia 1.15 g / l. Excellent tolerance. The patient notes an anorectic effect of the product.

The results of the clinical study show that:
1. The medicament of the invention is well tolerated clinically.

2. The biological tolerance is satisfactory the product has no detectable hematological, renal or hepatic repercussions.

3. The product lowers blood sugar in the diabetic subject without, however, putting the patient at risk of hypoglycemia when used as the sole treatment for diabetes. It is a true anti-diabetic.

4. Clinical efficacy is excellent in the treatment of non-insulin-dependent diabetes in adults and the elderly (obese diabetes, fiftieth diabetes, bloated diabetes); in addition to insulin therapy in insulin-treated diabetes, unstable diabetes, insulin-resistant diabetes.
The product is also effective in the treatment of overweight, in the treatment of hyperlipemia (hypercholesterolemia, hypertriglyceridemia); it exerts an anorectic effect in cases of overeating with overweight.

 Finally, it has been shown to be effective in the treatment of cataracts, in particular diabetic cataracts. This latter activity is entirely original of the medicament of the invention.

Claims (6)

1. Plant extract, characterized in that it contains a mixed polyphenolic and sterolic complex prepared from seeds or barks of Eugenia Jambolana Lamarck or other similar varieties of Eugenia or inner bark (cambium) of Anacardium Occidentale L, by treatment with water under heating of the ground starting material in order to remove the undesirable water-soluble fractions, filtration of the suspension obtained, then extraction of the filter cake using an organic solvent first of all cold then under heating, filtration of the insoluble material and dry evaporation of the organic solvent to obtain the extract containing the mixed polyphenolic complex and sterolic.  2. Plant extract according to claim 1> characterized in that the organic solvent is a lower alkanol, in particular ethanol.  3. Plant extract according to claim 1 or 2, characterized in that it has two characteristic absorptions in ultraviolet at 277.5 and 226 mA (El'lm / = 1.012 and 0.801 respectively), characteristic Rf at thin layer polyamide chromatography with ternary eluent butanol / acetic acid / water (30/55/15) of 0.22, 0.35, 0.54, 0.88 and 0.96.  4. Medicinal product having in particular antidiabetic, hypoglycemic, lipid-lowering, slimming, anorectic and cataract activities, characterized in that it contains, as active principle, a plant extract according to claim 1. 5. Medicament according to claim 1, characterized in that it is presented in a form suitable for oral administration, the active principle being optionally associated with pharmaceutical excipients.  6. Medicament according to claim 4 or 5, characterized in that it is presented in the form of unit doses containing from 0.5 to 3 g of active principle.