WO2003030919A1

WO2003030919A1 - Method of obtaining an extract of ginger

Info

Publication number: WO2003030919A1
Application number: PCT/CU2002/000008
Authority: WO
Inventors: Saulo GÓMEZ FERREIRA; Juana Irene TILLÁN CAPÓ; Yanier Núñez Figueredo; Sonia OLMEDO DE GÓMEZ; Sara María AGÜERON HERNÁNDEZ; Carmen CARRILLO DOMÍNGUEZ; Dayamí LAZA LOACES
Original assignee: Centro De Investigación Y Desarrollo De Medicamentos.
Priority date: 2001-10-11
Filing date: 2002-10-07
Publication date: 2003-04-17
Also published as: CA2463496A1; MXPA04003406A

Abstract

The invention relates to the pharmaceutical and food industry and, in particular, to a method of obtaining an extract of ginger by means of heat using butter as an extraction substance. The inventive method is used to produce a yellow ochre compound, of liquid and dense consistency at ambient temperature, which contains the principal fat-soluble components of ginger. Said compound has a therapeutic activity which has been demonstrated in laboratory animals in relation to acute, chronic inflammation and can be used as an antihistamine, an antiserotoninic agent and an inhibitor of passive cutaneous anaphylaxis (type I hypersensitivity reaction), induced by IgE, in order to treat, in particular, allergic asthma.

Description

PROCEDURE FOR OBTAINING A GINGER EXTRACT.

Technical Sector

The present invention relates to the Pharmaceutical and Food Industry, and in particular refers to a process for obtaining a ginger extract by heat, using butter as an extractor. An extract with proven therapeutic activity is obtained in experimental animals, which can also be used as a dietary supplement. Prior Art Ginger, or Zingiber official Roscoe (Zingiberaceae), is a plant native to Asia. Among its most significant uses is its use against diarrhea, vomiting, gastralgia, indigestion, whooping cough, fever, flu, cold, bronchitis, rheumatism, ulcer, cancer. Its use as an antibacterial has also been found. Butter is a natural dairy-derived product, composed of 80% fat, of which 50% is saturated, 25% unsaturated, 3% polyunsaturated, and 20% water in the form of dispersed drops containing 2% of proteins and salts. Clarified butter is known as Ghee, an "ancient food" that has been inscribed in the ancient Vedas religious books, which is considered a superior food with therapeutic properties. It does not decompose with the passage of time, maintaining its properties within which the increase in memory, increase in longevity, ^ improves circulation, as well as depressive, psychic and organic states, strengthens the immune system, repairs Damaged tissues, controls the excess weight of people with high cholesterol and fights bacterial and viral infections. US Patent 5,693,327 refers to the use of butter for making creams and ointments for the treatment of skin diseases. The procedures and equipment used for the extraction of ginger essential oils are described in US 4,257,945 and US 4,406,745. Its application in food preparation has also been protected. Patents US 4,206,241, US 5,447,735, US 5,468,493 and US 5,525,340 relate their use to the preparation of seasonings and food components, as well as flavorings of sweets low in cholesterol and fat, according to US Patent 6,235,334. More recently, US Patent 6,051, 235 also promotes its use in food manufacturing. In addition, combined with other plants, it is shown as a dietary supplement in US Patent 5,248,503; In this case, its usefulness is considered for the treatment of gingivitis, acne, colds and herpes simplex, among others, recommending various forms and routes of administration.

The action of ginger and other plant extracts on the enzyme 5-lipoxygenase, demonstrating its antioxidant activity, is reported in US 6,261, 607 and US 6,242,012. On the other hand, the action of ginger on the inflammation of bones and joints by inhibition of the enzyme cycloxygenase, which in turn prevents the synthesis of prostaglandins and leukotrienes, both mediators of inflammatory processes, is disclosed in US Patent 6,264,995, citing It also has antioxidant action to reduce the formation of free radicals. US Patent 5,494,668 shows a composition with ginger and other compounds, for the treatment of musculoskeletal diseases, since its inhibitory action on histamine release is known, by inhibiting both the lipoxygenase enzyme system and the cycloxygenase system. The process of extracting ginger with organic solvents shows an anti-inflammatory and antiplatelet activity in US Patent 6,274,177. Disclosure of the Invention. The present invention describes the natural process of extracting ginger using as an extractive substance a dairy compound of fatty composition known as clarified butter. An extract of yellow ocher color and of a liquid and dense consistency at room temperature is obtained, in whose composition the main fat-soluble ginger compounds are found and in which, the fundamental characteristics are maintained.

In the process of the invention, the fat soluble ginger compounds are extracted using clarified butter.

The clarification of the butter is done to remove the water and solid waste from it, and during the process a foam is generated that must be removed until it is no longer generated. For this, a temperature between 80 to 100 ° C is applied to the butter to be used for a period of 10 to 15 minutes. It is then filtered to remove any solid residue that may have remained.

The extraction process is carried out by mixing the clarified and filtered butter with clean and fresh ginger roots cut into thin slices of 2 to 3 mm thick in a proportion of approximately 0.5 - 8: 1 - 1.5, subsequently boiling said mixture during 20-30 minutes, until the ginger slices are dry. The extract thus obtained is filtered through a sieve between 2 and 2.5 mm, and if necessary it is filtered again to eliminate any present residue. A dense liquid of ocher yellow color is obtained. The extract obtained by the proposed process exhibits an anti-inflammatory, antihistamine and antiserotoninic effect by blocking the inflammatory action induced by carrageenan, histamine and serotonin in the rat's paw (Figure No. 1). It is observed that said extract has a marked antiserotoninic and antihistamine effect, since in the two inflammations where serotonin was present, it managed to reduce inflammation by more than 50%; and in the case of histamine the results were similar to that achieved by diphenhydramine. Ginger extract also showed an inhibitory effect of the IgE-mediated or conditioned hypersensitivity reaction (type I allergy) by inhibiting passive cutaneous anaphylaxis in rats (Figure No. 2) and effective antihistamine activity in the prevention of histamine-induced bronchospasm (Figure No. 3). It also has proven anti-arthritic effect by decreasing chronic inflammation induced by Freund's adjuvant (Figure No. 4)

Accordingly, the present invention also has as its object a pharmaceutical composition comprising as an active principle a ginger extract in clarified butter and an appropriate excipient, which can be in the form of a soft gelatin capsule, liquid formulation, emulsion and semi-solids. The invention also includes the use of ginger extract for the prophylaxis and treatment of allergic asthma, allergy induced by dusts, fungi, bites, hairs and skins of animals, atopic dermatitis and allergic rhinitis. The extract of the present invention can also be used as a nutritional supplement. EXAMPLES OF REALIZATION.

Example 1: Obtaining the ginger extract.

To obtain the clarified butter, 1 kg of butter is heated to 90X for 15 minutes, at which point the foam that has been generated is removed. Allow to cool to 60X and filter.

Starting 800 g of fresh, clean ginger roots, cut into thin films approximately 2.5 mm thick, it is added to the clarified and filtered butter, and is subjected to a heating process until it reaches 100 ° C for 30 minutes until the movies are dry. Finally, the extract obtained from the previous step is subjected to a filtration process through a 2 mm sieve until all solid waste is removed. The product obtained by this process is an oily extract of Zingiber offícinale Roscoe (Zingiberaceae) made from the rhizome of the fresh plant, in a material / solvent material ratio of 0.8: 1. Example 2: Test for anti-inflammatory activity in acute edema induced by phlogistic agents such as carrageenan, dextran, histamine and serotonin.

Phlogistic agents: histamine acid phosphate (BDH), serotonin sulfate (BDH), dextran (Cuba) and carregenin (Sigma). Test substances: MVZ, diphenhydramine (Medsol), cyproheptadine (Medsol), indomethacin (Medsol).

Experimental animals Female Wistar rats with a body mass between 150 and 200g were used, which were kept in a room with a controlled temperature of 22 ± 2 ° C, a bed with a chip with change every 48 hours and a light-dark cycle of 12- 12 hours..

Each group was made up of 6 animals which were individually identified for the exact dosage according to their body weight. The products were administered orally one hour before the subplantar injection of 0.1 ml of the phlogistic agents inducing the different edemas caused, which were inoculated in the right leg of the posterior train and 0.1 ml of saline solution (N _to CI ) 0.9% in the collateral leg. Table 1 shows the treatment schemes. Table 1. Scheme of treatment of edema induced by the different phlogistic agents

The increase in the volume of the leg treated with the phlogistic agent in relation to the leg against the side used as a control was measured on a Ugo Basile plethysmograph.

The results obtained were statistically processed by means of an analysis of variance of a classification route and later by a Duncan test. Figure No. 1 shows the anti-inflammatory effect of ginger extract, administered orally at the dose of 6.5 ml / kg. It presents the values in percent of inhibition of inflammation for each of the treated groups. The action of the extract is demonstrated by comparing its effect with the different substances used positive control, indomethacin, cyproheptadine and diphenhydramine, with which there is no significant difference. Example 3: Test of passive cutaneous anaphylaxis (APC). Type I hypersensitivity reaction.

For the passive cutaneous anaphylaxis assay, 1 mg ketotifen tablets were used as a positive control, lot 020899043, from Laboratorios Eron SA. Animals

For the test of passive cutaneous anaphylaxis, male istar rats of 300 to 350 g of body weight and male albino mice of 18 to 22 g of body weight were used, the latter for obtaining the ovalbumin antiserum. All animals remained in conditions of controlled temperature of 22 ± 2 ° C and light-dark cycle 12/12 hours.

For the test, work groups of three animals each were formed. One group received the ginger extract at a dose of 6.5 mL / kg, the positive control group administered with ketotifen 3mg / kg and a control group that received no treatment. Rats were shaved in the dorsal area 1 h before intradermal administration of 0.1 mL of egg albumin antiserum at 6 loin sites. After 24 h, the anaphylactic reaction was caused by intravenous administration of 1 mL of a 0.1% solution of egg albumin in the 1% Evans blue dye. The animals were sacrificed, 1h after the anaphylaxis reaction was carried out, skin portions were removed and placed in test tubes containing 4 mL of formamide and incubated at 37 ° C for 48 h. The optical density of the extravased dye was measured at λ = 623 nm. The micrograms (μg) of extravased Evans blue / site were obtained from a dye calibration curve. The results obtained were processed by a one-way analysis of variance and then a Duncan test was performed with a significance level of p <0.01.

The results are shown in Figure No. 2, which shows an inhibitory effect of passive cutaneous anaphylaxis of ginger extract comparable to ketotifen, as there is no significant difference between them. A decrease in the amount of extravasant dye / site (μg / site) is observed in these two groups in relation to the untreated negative control group, reflecting the inhibition of the anaphylaxis reaction typical of type I hypersensitivity reactions present in the processes of allergic asthma. Example 4: In vivo antihistamine activity test. Inhibition of histamine-induced bronchospasm.

For the in vivo antihistamine test, male Hartley guinea pigs (400-450 g body weight) were used. Test groups of 6 animals were made, one treated with the ginger extract 6.5 mL / kg administered orally for 7 days and a control group that received no treatment.

Subsequently, all animals were anesthetized with 40 mg / kg of sodium pentobarbital in order to perform the tracheotomy and ventilate them with a breathing pump; the jugular vein was channeled and 15 mg / kg of sodium pentobarbital were administered to produce respiratory arrest, then 3 intravenous administrations of 15 μg / kg of histamine were made to both the treated group and the control group and the pressure was recorded bronchial after each administration. The result is considered positive when the average of the values of the difference in bronchial pressure of each animal caused by the administration of histamine in animals treated with ginger extract turns out to be less than 50% of the average of the values of the difference of bronchial pressure of control animals. 100% of the animals administered with the ginger extract composition in butter orally over a period of 7 days, showed effectiveness in the prophylactic treatment of histamine-induced broncho-constriction by meeting the positivity criteria. In addition, the values of the control group and the treated group differ significantly according to the statistical criteria analyzed, this effect is mediated by the inhibitory action of ginger extract on the effects of histamine at the bronchial level (Figure No. 3).

Example 5: Test of antiarthritic activity. Chronic inflammation induced by Freund's adjuvant. The effect of ginger extract on Freund's adjuvant-induced arthritis was tested.

Rats distributed in three treatment groups were used for the study. A first group treated with tenoxicam as a positive control at a dose of 2.5 mg / kg, another group that was treated with ginger extract at a dose of 6.5 ml / kg. and a third who did not receive treatment All animals were administered in the right leg of the back train 0.05 ml of complete Freund's adjuvant and at 18 days the administration of ginger extract and tenoxicam was started orally during a period of 12 days. At the end of the administration, the Increase in the volume of the leg treated with the complete Freund's adjuvant in relation to the counter-lateral leg used as a control, in a plethysmograph.

The results are shown in Figure No. 4. It is observed that ginger extract significantly inhibits inflammation induced by Freund's adjuvant, its effect being comparable to that achieved with a dose of 2.5 mg / kg of

Tenoxícam, as there is no significant difference between them.

Brief Description of the Figures:

Figure No.1: Effect of ginger extract (EJ 6.5 mL / Kg) on edema induced by different phlogistic agents (anti-inflammatory activity). Figure No.2: Effect of the compound on passive cutaneous anaphylaxis in rats

(APC reaction inhibition). Uneven letters differ significantly for

(p <0.01).

Figure No.3: Effect of the compound on bronchoconstriction caused by histamine ("in vivo" antihistamine activity). They differ significantly (p> 0.01). Figure No.4: Effect of M.V.Z. in Freud's adjuvant-induced chronic edema

(antiarthritic effect). Equal letters differ significantly (p <0.01).

Claims

1. Procedure for obtaining a ginger extract with pharmacological activity characterized in that butter is used to extract fat-soluble compounds from ginger.

2. Method according to claim 1 characterized in that for the extraction of fat-soluble compounds of ginger, thin films of ginger roots are mixed with butter, previously clarified and filtered, in a proportion 0 ginger / butter of approximately 0.5-8: 1-1. 5, boiling said mixture for 20-30 minutes and finally filtering the obtained extract.

3. Ginger extract obtainable by the procedure of claims 1 and 2.

4. Ginger extract characterized in that it is obtained by extracting 5 fat-soluble compounds from ginger using butter.

5. Ginger extract according to claims 3 and 4 characterized in that it is an oily yellow oily extract with a liquid and dense consistency at room temperature.

6. Pharmaceutical composition characterized in that it comprises as principle 0 active an extract of ginger in butter and an appropriate excipient.

7. Pharmaceutical composition according to claim 6 in the form of a soft gelatin capsule, liquid formulation, emulsion and semi-solids.

8. Use of the ginger extract according to claims 3 to 5 as an antihistamine. 5

9. Use of the ginger extract according to claims 3 to 5 as an antiserotonin.

10. Use of the ginger extract according to claims 3 to 5 as an anti-inflammatory.

11. Use of the ginger extract of claims 3 to 5 as an antiarthritic.

12. Use of the ginger extract of claims 3 to 5 for prophylaxis of histamine-induced bronchospasm.

13. Use of the ginger extract of claims 3 to 5 for the prophylaxis and treatment of the hypersensitivity reaction mediated or conditioned by IgE.

14. Use of the ginger extract of claims 3 to 5 for the treatment of type I allergy.

15. Use of the ginger extract of claims 3 to 5 for the prophylaxis and treatment of allergic asthma.

16. Use of the ginger extract of claims 3 to 5 for the prophylaxis and treatment of allergy induced by powders, fungi, bites, hairs and skins of animals.

17. Use of the ginger extract of claims 3 to 5 for the prophylaxis and treatment of atopic dermatitis.

18. Use of the ginger extract of claims 3 to 5 for the prophylaxis and treatment of allergic rhinitis.

19. Use of the ginger extract of claims 3 to 5 as a nutritional supplement.