DE2407910A1 - PROCESS FOR EXTRACTION OF TANNINS FROM FRUITS OF CUPRESSUS SEMPERVIRUS AND TANNIN-CONTAINING PREPARATIONS OF THE COSMETIC AND PHARMACEUTICAL INDUSTRY - Google Patents

Description

23 (P-8)

DR. ING. BRIGITTE NÖTHE 8019 Ebersberg 1 FEB 9. Wk

,. " _{,, ^ 1 w} , · Abt-Williram-Sttmo 25

- PATENT ADVERTISER Tefcfon * 8 10 (area code 08092)

Yves Andre VINCENT
Marseille (France)

Process for the extraction of tannins from fruits of the Cupressus sempervirens and preparations containing tannins of the cosmetic and pharmaceutical industries

The invention relates to a method of extraction of tannins based on fruits of Cupressus sempervirens as well as tannin-containing products or tannin preparations of the cosmetic and pharmaceutical industries, which are obtained on the basis of these extracts.

Tannins are known to be widespread in the plant kingdom and can be obtained from tree bark (e.g. from oaks and mangroves), Wood (oak, chestnut, quebracho etc.), roots (especially oak), leaves (sumac), or fruits (gonakia, Dividivi, Myrobalanen, etc.) can be extracted. Their ability to keep skins from rotting is well known. The tannin-containing mass that can be extracted from the fruits of Cupressus sempervirens is made by a mixture of more or less condensed or agglomerated molecules are formed, of which the tannins are of low molecular weight

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(Molecules with less than 5 elementary units or "motifs") are unsuitable for tanning hides, while those with 5 to 10 elementary units develop tanning effects; in addition, they are no longer soluble.

Some tannin preparations are also known for their therapeutic properties, especially as astringents, which are equipped with a counteraction against diarrhea and hemorrhages. We also know that the fruits of the common Cypress or Cupressus sempervirens, which is called a plump Cones and in France under the name "Galbules" are known to contain tannins, which have long been used in human therapy for the treatment of venous disorders. According to the state of the art, these tannins are obtained as total extracts of the fruits of Cupressus sempervirens. These however, known extracts have only moderate therapeutic effectiveness.

Surprisingly, however, it has now been found that the therapeutic effectiveness of tannin extracts from fruits of Cupressus sempervirens through well-coordinated manufacturing methods of preparations with highly purified tannins or strong enriched tannin fractions compared to the known total tannin extracts can be increased extraordinarily.

On the one hand, the applicant could use a fractionation the tannin-containing mass obtained by total extraction of the fruit of Cupressus sempervirens surprisingly determine that the weakly condensed tannins have little or no effect on the veins of mammals, while the strongly condensed tannins have a very significant physiological effect on the venous system.

On the other hand, the applicant has found that the content of

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Fruits of Cupressus sempervirens on strongly condensed Tannins can vary considerably depending on the season in which the fruit is harvested can become garbage. The applicant was able to determine that the content of strongly condensed tannins increases regularly, when the young fruit grows from the minimum size to a weight of 5 g, which takes about 3 months (from March to June in the south-east of France). Then the content remains constant from the 4th to the 8th month, while the fruit increases from 5 to 10 to 12 g (from July to October in the south-east of France). After that, the content of condensed increases Tannins decrease rapidly and become practically zero around the 10th to 11th month (January to February in the southeast of France).

The extracts of cypress fruit thus have maximum therapeutic value when made in summer while the extracts made with fruits harvested in winter are very low, if any have therapeutic value. The content of condensed tannins in the cypress fruits appears according to the temperature profile and even more likely to follow the rhythm of the renewal and refreshment of the juices. The applicant has in the fat also found that the over the 12 · month Fruits remaining on the tree when they resume or when the sap rises again in the spring condensed again Loaded tannins, but to a lesser extent than during the initial cycle.

In this context, it was also found that the fruits collected in the 6th month when stored in a cold room of + 5 ° C under suitable conditions to avoid the formation of mold and drying out their content keep on condensed tannins for a long time. If you have the

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If, on the other hand, the same fruits are left in the air at ambient temperature, they will open, lose or loosen their seeds and dry out, after which no more strongly condensed tannins are found in it.

The invention therefore relates to a method for extracting tannins from fruits of Cupressus sempervirens, with which tannins can be obtained, their therapeutic effectiveness compared to known methods obtained tannin extracts is noticeably increased, so that therapeutic effects of great effectiveness with the new preparations can be obtained at minimal doses of active ingredient.

The method according to the invention suitable for this purpose is characterized in that the fruits of the Cupressus sempervirens harvested between the 4th and 8th month after flowering or the fruits of the Cupressus sempervirens that have remained on the tree and harvested between the 16th and 20th month after flowering are used subjected to multiple extractions with an aqueous solvent; the total extract obtained, possibly after previous evaporation or concentration until a powder is obtained, treated with about 90 full (90 ⁰ GL, ie 90 Gay-Lussac degrees) alcohol or any other suitable solvent to separate the therapeutically inactive, weakly condensed tannins, which are soluble in said alcohol or other suitable solvents; and the remainder of the strongly condensed tannins, which are freed from weakly condensed tannins and which are insoluble in the alcohol, is obtained as a therapeutically active fraction of the tannin extracts from the fruits of Cupressus sempervirens.

According to a further embodiment of the method according to the invention, the data between the 4th and 8th month or possibly Harvested between the 16th and 20th months after flowering

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Fruits of Cupressus sempervirens are used either immediately after harvest or after a longer storage time, which can reach several months, whereby the storage should take place at a temperature in the region of + 5 ⁰ C under conditions that are suitable for both mold formation and dehydration to prevent.

The invention therefore relates to tannin preparations, obtained from the fruits of Cupressus sempervirens by the process mentioned and by strongly condensed, roughly 90 complete (90 ° G.L.) Alcohol insoluble but water-soluble Tannins are formed.

The invention also includes industrial applications of these preparations. In particular, it includes new medicaments which can be used both in human medicine and in veterinary medicine, which are formed as active ingredients by the strongly condensed tannins of the fruits of Cupressus sempervirens or contain such and especially therapeutic agents for the treatment of venous diseases.

In the same way, the invention also includes cosmetic preparations that contain strongly condensed tannins which are obtained from the fruits of Cupressus sempervirens.

Further special features will emerge from the following description.

Illustrative examples for the preparation and pharmacological testing of the tannin extracts according to the invention follow.

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Example 1 (extraction of the tannins)

1000 kg were used between the 4th and 8th month appropriately washed fruits or cones of cypresses harvested after flowering. The fruit turned into pieces crushed by a few millimeters, which immediately in a container with 2000 l refluxing, about 50 complete (50 ° G.L.) Were given ethyl alcohol to inactivate the enzymes. The mass was after the Extractor kept boiling for another 0.5 hours. Then became (n) the heating turned off and the 2,000 1 alcohol removed and replaced by fresh 2,000 1 50 tfigen alcohol that brought to the boil was brought and held for 0.5 hours. After cooling, the extract was drawn off and combined with the first extract.

The 4000 l alcoholic extract were prevented from taking Pressure from 20 to 40 Torr at a pressure not exceeding 50 ° C Temperature evaporated to such an extent that about 400 l remained. The concentrated extract obtained in this way became insoluble Resins freed and carefully filtered and then dried in a vacuum dryer or better by atomization. on in this way 70 to 85 kg of composite tannins in powder form were obtained.

These composite or complex tannins were dispersed in 5 volumes (based on the weight) of about 85 complete (85 ⁰ GL) ethyl alcohol and the dispersion was stirred for 2 hours at room temperature. The insoluble fraction was then separated off on the filter, the cake being washed with fresh approximately 90% (90 ° GL) alcohol. The suctioned or spun off filter cake was divided in a granulator and dried in vacuo at a temperature which

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should not exceed 50 ° C, spread over plates. the dry granules were then crushed to the desired grain size. In this way, 25 to 35 kg were condensed Preserved tannins.

In the example above, ethanol was used as the extractant for the total tannins, but it is clear that any other solvent such as in particular methanol, isopropanol, acetone, methyl ethyl ketone, etc. mixed with water can be used in appropriate proportions.

Example 2 (pharmacological tests)

On the basis of pharmacological investigations in which vein motility tests were carried out, it could be demonstrated that the little condensed tannins extracted from the fruits of Cupressus sempervirens, ie those that ^{are soluble in about 90% by volume (90 0} GL) alcohol, have practically no effect exercise on the veins of mammals, while the highly condensed tannins, which are isolated according to the invention and are both ^{insoluble in about 90 vol.% (90 0} GL) alcohol and soluble in water, a significant ·? Tend physiological effectiveness, which is very much higher than that of the total tannins.

Hind legs of the frog (Rana esculenta) and proceed as follows:

When perfusing with Ringer's fluid via a cannula connection in the section of the previously isolated lower aorta under constant pressure (about 15 cm water column over or Differential pressure) and recovery of the perfusate via the

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If the inferior vena cava was isolated, the flow rate was measured with a photoelectric drop counter. Over a Branching, known amounts of vein-constricting products could be injected into the aorta, resulting in a decrease of the perfusion throughput. After the product injected into the perfusate has emerged or left, the throughput should return to normal. The aim was, by means of a few experiments (which were carried out as quickly as possible, in order to not to tire the preparation) to determine the dose that is proportional to a temporary decrease in flow 1: 5 leads. The experiment was repeated on at least 10 animals. According to this technique, it was found that on average 40 ug highly condensed tannins or 200 to 300 ug total tannins (depending on their composition) are required to reduce the flow to e ± a 1/5.

Soluble in about 90 complete (90 ° G.L.) Alcohol weakly condensed tannins, it is not possible to decrease the flow to 1/5 because of injections of 1000 µg (1 mg) of weakly condensed tannins a maximum reduction to 1/2 is achieved. Higher doses lead to irreversible changes in the preparation and are therefore no longer physiologically sensible.

These results show the high effectiveness of the heavily condensed tannins as opposed to the very low ones Effectiveness of the less condensed tannins, which also have a pronounced inhibitory effect when used with highly condensed tannins remain mixed (as is the case with the whole tannin extract). Hence the need arises to remove the weakly condensed tannins, if one arrives at an improved therapeutic prognosis want.

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-9- 2A07910

The strongly condensed tannins obtained according to the invention are for preparations galenic type, as they are in the Human medicine or veterinary medicine can be used and can be used directly for addition to cosmetic products as well For all commercial applications that require highly purified leucocyanic tannins.

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Claims (5)

Claims V A method for obtaining tannin extracts from fruits of Cupressus sempervirens, characterized in that between the 4th and 8th month after flowering or the remaining on the tree and harvested between the 16th and 20th month after flowering fruits of the Cupressus sempervirens is subjected to one or more extractions with an aqueous solvent and that the total extract obtained, if necessary after evaporation until a powder is obtained, with about 90 vol.tigern (90 ⁰ GL) alcohol or any other suitable solvent to separate the therapeutically inactive solvent therein Treated weakly condensed tannins, while the heavily condensed tannins insoluble therein are collected or obtained as a therapeutically effective fraction of the tannin extract of the fruits of Cupressus sempervirens. 2. The method according to claim 1, characterized in that the between the 4th and 8th month or optionally between the 16th and 20th month after flowering harvested fruits of Cupressus sempervirens immediately or after storage for several months at a temperature in the area of + 5 ⁰ C under suitable conditions to avoid mold formation and drying out. 3. The method according to claim 1 or 2, characterized in that the particular crushed fruits of Cupressus sempervirens an extraction with refluxing about 409847/1 1 17 Subject to 50 total (50 ° GL) ethanol; the extract obtained is preferably concentrated under reduced pressure and at a temperature not exceeding ^{50 ° C .;} separating the concentrated extract from insoluble resins; the composite tannins thus obtained are ^{dispersed in about 85 vol.tigern (85 0} GL) ethanol and the dispersion is stirred at room temperature for an appropriate period of time in the region of 1.5 to 3 hours and then the insolubles are separated off by filtration as a filter cake, which is mixed with fresh , about 90 vol.tigern (90 ⁰ GL) ethanol is washed and finally preferably ^{dried in vacuo and at a temperature not exceeding 50 0} C to obtain in about 90 vol.tigern (90 ⁰ GL) alcohol insoluble, water-soluble, highly purified , strongly condensed tannins. 4. Strongly condensed tannins obtained according to one of the preceding Expectations. 5. Tannin-containing or tannin preparations of the cosmetic and pharmaceutical industry, characterized by strongly condensed Tannins according to claim 4 as an active ingredient.