FI103258B - Process for the preparation of bioactive products derived from two rv - Google Patents

Description

5, 103258

Method for the production of bioactive products derived from peat - Torv - Förfarande för framställning av bioaktiva produkter härrörande frän Torv

The present invention is primarily directed to processes for preparing bioactive products derived from peat. The invention also relates to methods for preparing pharmaceutical and / or cosmetic formulations or compositions containing said peat-derived bioactive products.

Extraction of peat by various methods using various extraction media and the use of such extracts containing peat-derived bioactive ingredients for cosmetic and pharmaceutical purposes is known.

These known methods are described in Polish Patent Specification No. 124,110 (Chemical Abstracts 101 (10), 78854e).

Bioactive products derived from peat by this prior art process are obtained by primary and secondary early hydrolysis of the air-dried raw peat material, followed by acidification of the hydrolyzate thus obtained and subsequent separation of the insoluble solids by a second alkalization, clarification with a clear liquid phase, and by ether extraction. In said process, the aqueous phase derived from the organic extraction is a liquid bioactive product derived from peat.

t. The known liquid product, which is a solution of the active ingredients derived from peat in a highly concentrated, almost saturated aqueous solution of sodium chloride and obtained according to the above-mentioned Polish Patent Publication No. 2,103,258, is unstable when stored for a long time and also contains the biological activity of the composition. a large excess of neutral inorganic substances. As a bulk product, it is difficult to handle, store and process.

The main object of the present invention is to provide a product which does not have these disadvantages, i.e.. a product which is stable and can be formulated readily in pharmaceutical and veterinary medicine and can be incorporated either in solid form or in any suitable solution in cosmetic products.

In particular, the use of a bioactive product derived from peat for pharmaceutical purposes, i. the preparation of pharmaceutical compositions requires a solid form which is well suited for this purpose. Since previous attempts to concentrate and desalinate known aqueous solutions of peat-derived bioactive substances have not produced a positive result due to the inclusion of the active ingredients in the crystallization solid phase, resulting in a decrease in the biological activity of the composition, it was difficult to find a suitable powder form.

Unexpectedly, it was found that a positive result can be achieved if the liquid composition is first diluted several times prior to concentration, i. volumes of water, which are many times the volume of the composition.

Thus, the present invention provides a process whereby a peat-derived bioactive product having the above advantageous characteristics is obtainable from a highly concentrated aqueous solution of an inorganic salt, especially sodium chloride, containing the peat-derived bioactive ingredients by diluting said solution with water, : desalted, eg with distilled water, followed by reverse osmosis, concentration and clarification.

3 103258 The solution thus obtained can be easily transformed by sterilization and spray drying into a sterile and solid product which is well suited for the intended purposes. The dilution is preferably carried out with amounts of water several times, preferably 5-8 times, the amount of concentrated solution to be diluted.

This process, which is used to treat the product obtained according to Polish Patent No. 124,101, consists of separating the remaining organic solvents from the post-extraction aqueous phase, separating the insoluble parts by filtration under reduced pressure through sintered ceramic material, diluting several times with water, to separate excess mineral salts, mainly sodium chloride, as permeate. The desalted solution is then concentrated, clarified by centrifugation, and sterilized by filtration through a membrane filter, e.g., a Millipore (r) -20 filter. The microbiologically pure solution obtained may optionally be spray dried. The sterilized product (liquid or solid) may be formulated as a cosmetic or veterinary composition. The concentrated and clarified solution may be used optionally without sterilization and without spray drying as any suitable dilution component in a variety of cosmetic compositions.

In the spray drying step, the inlet temperature is preferably set at about 180 ° C, while the outlet temperature is set at 30 to about 90 ° C.

Although the process of the present invention is described above in combination with the process of Polish Patent No. 124,110, its use is not limited to such a combination, but is generally available in the process of obtaining a bioactive product of peat from a highly concentrated aqueous solution of inorganic salts, especially sodium chloride. , which contains bioactive ingredients derived from peat.

103258 4

Most preferably, the peat-derived bioactive products provided by the present invention do not contain more than 70% by weight, preferably more than 60% by weight, of inorganic salts, in particular sodium chloride. Because as low a concentration of sodium chloride concentrate as possible is desired for optimal product, especially for pharmaceutical applications, and for cosmetic uses requiring higher concentrations, i. for facial treatment, lower concentrations of sodium chloride, such as 55% or even less, are preferred, especially when obtained by the dilution and reverse osmosis steps.

If the process is terminated by the concentration and clarification steps, the product is a concentrated (or thickened) solution. "Concentrated peat extract" as used in this specification is a dark brown liquid having a density of 1.02-10.9 g / ml and a dry matter content of not less than 5% by weight. The chloride ion content in dry solids, calculated as NaCl, does not exceed 70%, preferably 60%, and the pH of a 1% aqueous solution is 5.0 to 6.5, usually about 6.0. The absence of an additional sterilization step is not a list in certain cases, e.g., certain cosmetic uses of the concentrated peat extract in question.

On the other hand, a sterilization step is mandatory if the product is intended for the preparation of pharmaceutical compositions. Particularly in such a case, the additional step of spray drying is most preferred, though not mandatory. The product obtained after such a spray-drying step is in powder form and is therefore particularly suitable for the preparation of certain pharmaceutical compositions. The most preferred product of this type is the product commercialized under the name "TOLPA (R) Torf Preparation" with TOLPA (R) being Torf Corporation, ui. Mydlana 2, Wroclaw, Poland, is a registered trademark. This abbreviation uses the abbreviation Tässä to describe the product.

The present invention also relates to processes for the preparation of pharmaceutical compositions containing, as active ingredient, the bioactive product derived from peat 5103258 described above, in particular the product containing not more than 70% by weight, preferably more than 60% by weight, of inorganic salts, in particular sodium chloride, calculated on the dry solids, together with a pharmaceutically acceptable carrier.

The peat-derived bioactive product incorporated into such pharmaceutical compositions is preferably TTP as defined above. The pharmaceutical preparation contains a bioactive product derived from peat and a pharmaceutically acceptable carrier material, preferably in a weight ratio of about 1: 5 to 1.25 and most preferably 1: 9 to 1:19.

The present invention is further directed to a process for the preparation of a pharmaceutical formulation containing a peat-derived bioactive product in gel form. This method is characterized in that sterile alcoholic herbal extract is combined with sterile glycerol, a sterile, preferably concentrated, aqueous and sterile menthol solution of a previously formulated peat-derived bioactive product, and that the resulting mixture is gradually mixed with : 10-94: 6. TTP is preferably used as a powdered or concentrated bioactive product derived from peat.

The present invention also relates to a process for the preparation of a pharmaceutical formulation in the form of a cream containing a bioactive product derived from peat. Such a process is characterized in that the sterile herb extract is gradually combined with a sterile solution of a peat-derived bioactive product in powder form, the resulting mixture is gelled by the addition of colloidal silica, and the gel thus obtained is previously triturated with 35 sterile fat components, the weight ratio of the liquid components to silica being about 30:20 and to the fat composition of the gel. 32: 68-34: 66. Here again, TTP is used as a powdered or 6 103258 concentrated bioactive product derived from peat.

Cosmetic preparations which may contain herb extracts and other auxiliary and enrichment components, fragrance compositions and carrier materials permitted for cosmetic use contain from 0.01 to 10% by weight, preferably 0, of a peat-derived bioactive product of the present invention. , From 05 to 1.00% by weight and preferably from 10 to 0.010% by weight.

The carrier materials may be aqueous alcohols, emulsions of all types, gels, foaming compositions, and fat carriers. The use of one specific carrier selected from the group of the above-mentioned substances allows the formulation of various types of cosmetic products of the invention, such as daily lotions, foams, balms, cleansing milks, etc., as well as shampoos, hair balms, bath foams, all bioactive products.

Peat-derived bioactive products (peat extracts in abbreviated form) derived from crude raw materials such as (including) medicinal mud contain well-balanced amounts of mineral and organic compounds such as B, Si, Ab, Fe, Mg, Mn, Cu, Sn, Ni, Ca, Ag and Na; organic compounds such as amino acids in free form and salts; polysaccharides, partially degraded / reacted during hydrolysis to deoxysaccharides and / or amino acid saccharides. Peat, especially medicinal mud, is known and identified as a material of biological plant and microorganism origin; due to the concentration of its nutritional and stimulatory components, it has beneficial effects on humans and mammals; thus, the bioactive compositions derived from peat contain the above-mentioned substances in proportions which are characteristic of living organisms; this is considered to be an explanation of the beneficial effects of cosmetic and pharmaceutical preparations containing 7,103,258 peat-derived active products and compositions. Particularly good effects of the new cosmetic compositions are observed when 5 herbal extracts are also present in the formulation. The selection of a suitable herbal extract is based on the known routine use of such extracts in cosmetics, which modifies the activity of the preparations and thus allows the contents of the cosmetic products to be consistent with the requirements and needs of the persons being treated.

The present invention will be further characterized and explained in the following examples.

Example 1

Starting with 1000 kg of air-dried raw peat material following, for example, the process according to Polish Patent No. 124110, a solution of peat-derived bioactive ingredients in saturated aqueous sodium chloride solution was obtained in an amount of about 10 liters. The solution was filtered through a sintered ceramic filter under reduced pressure to clarify the solution before desalting. The clear solution thus obtained contains about 95% NaCl in the dry mass. The dry mass is formed by about 32% by weight of the solution. This clear solution has a volume of about 7 liters.

The clear solution is diluted 5 to 8 times with distilled water and subjected, in diluted form, to a desalting step, which is performed using a reverse-moose technique using a DDS. Desalting is carried out for 3 to 4 hours, during which the excess of mineral salts, mainly NaCl, is removed in the form of permeate.

The desalted composition contained approximately 65-70% sodium chloride in solids. The solution thus obtained, 35 6-7 liters, was concentrated 4-5 times in a Buechi rotary evaporator to give a concentrated solution containing approximately 20% dry weight. The resulting concentr. the purified solution was clarified using a Biofuga-Heraeus sen- 8 103258 trifuge apparatus (flow separator) and then sterilized by filtration through a Millipore (R) filter.

The resulting microbiologically pure solution was spray dried in an Anhy-5 dro dryer with the outlet flow set to 90 ° C and the inlet temperature set to 180 ° C. The yield of the dried powder was approximately 200 g.

Example 2 The product obtained in Example 1 was used for the preparation of pharmaceutical gel and ointment compositions which also contain herbal extracts synergistically, which improves the therapeutic effect in some diseases. The anti-leg ulcer gel and ointment were prepared as follows: 15 g of horse chestnut extract, 10 g of marigold extract, 60 g of Gly serol, 0.1 TTP obtained in powder form as described in Example 1, 0.1 g of salicylic acid, 1.0 g of distilled water and 8.8 g of Aerosil (R) (colloidal silica) were used to obtain the gel form of the preparation.

20

The liquid (non-volatile) ingredients were sterilized by heating under reflux for two hours before use. The herbal extracts were combined with glycerol and an aqueous solution of TTP and menthol and silica were gradually added with continuous stirring.

Similarly, the following ingredients were used to obtain the ointment composition: 22 g of horse chestnut extract, 10 g of marigold extract, 0.1 g of salicylic acid, 0.1 g of TTP in powder form, obtained as described in Example 1, and 2.0 Aero-silica (R ).

«

The fat components used were a mixture of the following substances: 22 g euserine and 45.8 g petrolatum. The herbal extracts were sterilized by heating for about 2 hours under reflux. Euserine and petroleum jelly were sterilized in the same manner. The liquid ingredients were carefully combined with silica to give a gel which in turn was triturated with sterilized and fat components cooled to room temperature. A stable ointment was obtained which did not stand out on storage.

The above gel and ointment were used simultaneously to treat 5 leg ulcers. The wounds were treated with a gel formulation, while the surrounding intact skin was treated with an ointment. The addition of colloidal silica is believed to be responsible for proper drying, while the herbal and peat derived components are believed to be responsible for the healing effect of the preparation. The fat components helped keep the skin and skin elastic. The results obtained were compared with a control patient group treated in a classical manner. Those who received new treatment were selected from a group of patients who had suffered from the disease for 15 months (sometimes years) without significant positive effects. Patients treated with the compositions of the invention exhibited better results over the first few weeks than control patients.

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Example 3

The products obtained in Example 1 were used for the preparation of pharmaceutical formulations in the form of tablets or capsules.

25

The sterile peat-derived bioactive composition in powder form was combined with a carrier at a weight ratio of 1: 9. MYVATEx (R) TL (trade name of Eastman-Kodak), a mixture of lactose and lubricants, in a weight ratio of 44: 1 was used as carrier. The 50 mesh particle size lactose and MYVATEX (r) teas were finely ground so that approximately 70% of its mass passed through a 100 mesh sieve. A portion of the resulting mixture of the active composition and carrier was formulated into tablets containing 5 mg of the active ingredient. The total weight of each tablet was 50 mg. Another portion of the same mixture of active composition and carrier was granulated using q.s. ethanol (40% by volume). The granules were screened and ground as needed and filled into capsules in an amount such that each capsule of 103258 contained 5 mg of active ingredients, using TTP as a 19: 1 mixture with carrier.

The above tablets were tested in their artificial gastric fluid at 37 +/- 2 ° C to measure the disintegration time using an Erweka device. An artificial gastric fluid was prepared as follows: 2.0 mg of sodium chloride and 3.2 g of pepsin were dissolved in hydrochloric acid and distilled water was added to a total volume of 1000 ml. The resulting solution had a pH of about 1.2. The disintegration time of a tablet of 5.1 mm diameter and 0.0498 g total weight was 6 minutes.

Other examples are directed to a plurality of cosmetic formulations of the invention of various compositions and for various purposes containing advantageous additions of peat-derived bioactive agents. Various facial lotions, balms, creams, milks, shampoos, foam bath lotions, etc. are described.

Example 4:

To the reaction vessel equipped with a stirrer was charged 150 mg of chamomile extract obtained by extraction of chamomile lacquer with 1: 1 ethanol: aqueous solution and 1 g of the TTP described above. To the resulting mixture was added 50 g of glycerol. The three substances were mixed to form a uniform mixture. Next, another mixture formulated as before was introduced into the same reaction vessel. It consisted of 340 g of 95: 5 ethanol-30 L: aqueous solution, 1 g of salicylic acid and 0.5 g of menthol. The two mixtures were combined by stirring to obtain a uniform solution. Next, 3 g of the fragrance composition slurry TILIANA H4308 was added. TILIANA H4308 is a product of Fabryka Syntetykow Zapacho-Wych Pollena-Aroma (Syntetic Fragrance Works Pollena-Aro-35 ma), Warsaw, Poland. The solution was then brought to a total volume of 1000 ml by the addition of 454.5 g of distilled water; stirring was continued until a homogeneous mixture was obtained.

n 103258

The above procedure used 86% menthol and water according to Polish Pharmacopea FP IV and ethanol at 95% concentration according to Polish industrial standard BN-75 / 6193-01.

The concentrated peat extract used was a dark brown liquid having a density of 1.020 to 1.090 g / ml and a solids content of greater than 5%; The pH of a 1% aqueous solution was 5.0-6.5. The camomile extract was a reddish brown liquid having a density of 0.9160-0.9503 at 10 g / ml and an ethanol content of 52-56% by volume.

The face water preparation obtained above is suitable for all skin types. It is a clear liquid with no solids. Its color is yellow. The pH is 4.28 and the ethanol content is 15.45.92% by volume. The total acidity, calculated as salicylic acid content, was greater than 0.1% by weight, namely 0.23% by weight. The product did not lose any of the above characteristics during storage for 12 months.

Example 5

The procedure described in Example 4 was repeated with the only difference that the flowers and the FINISH H4625 (which is also a product of Fabryka Syntetykow 25 Zapachowych Pollena-Aromaj) were used in the same molar and weight ratios instead of chamomile extract and TILIANA H4308 fragrance composition. The resulting lotion is suitable for dry and sensitive skin. It was also a clear liquid with no solid particles. The pH was 4.30, the total ethanol content was 45.82% by volume and the total acidity was 0.27% by weight. When stored, the preparation remained unchanged for 12 months, as was the preparation obtained in Example 4.

Example 6

35 The procedure described in Example 4 was repeated except that sage leaf extract was chosen instead of chamomile extract and LELIA 90368 (Pollena-Aroma, Warsaw) was selected as TILIANA

t> H4308 instead. The sage leaf extract was obtained by extraction of the dried sage leaves with ethanol at 50 ° C and was, 2 103258 brown, having a characteristic sage scent and a density of 0.9503; it contained 52-56% ethanol. The resulting facial conditioner is particularly suitable for oily skin. It is a clear and homogeneous liquid with a dark yellow color; The pH, ethanol content and acidity were comparable to the conditioner of Example 4.

Example 7

The following composition was shown to be particularly effective in preventing or treating rapid tooth loss: 24.0 g of chamomile extract 3.0 g of sage leaf extract 0.3 g of salicylic acid 0.2 g of menthol

0.1 g TTP

Up to 100.0 g commercial gel base.

Example 8 The following carefully weighed components were introduced into a 200 ml reaction vessel equipped with a mechanical stirrer: 270 g of camomile extract obtained by extraction of camomile lacquer with 50% ethanol; the extract was a reddish brown liquid having a density of 0.9160-0.9503 g / ml and an ethanol content of approximately 55 volume%, 50 g glycerol, 86% according to Polish Pharmacopean FP IV, 30 g saponaria officinalis extract obtained by extraction -30 la saponaria officinalis in 70% ethanol; the extract was a tan liquid with a density of 0.9630-0.9810 g / ml and an ethanol content of approximately 75% by volume,.

1.0 g of the inventive concentrated peat extract, a dark brown liquid having a density of 1.02 to 1.09 g / ml, not more than 35 2% chloride ions calculated as sodium chloride; the dry solids content was above 5% ', the pH of the 1% aqueous extract was 6.0.

• »13 103258

The ingredients listed above were thoroughly mixed. A previously prepared solution of 1 g of salicylic acid in 260 g of 95% ethanol was added thereto. A combined solution of 383 g of distilled water and 5 fragrance compositions of 5 TILIANA H4308 was added and stirred until a uniform solution was obtained. The preparation was analyzed and stored in 200 ml retail bottles. The product obtained was suitable as a hair care product. It was a clear, slightly opalescent liquid containing approximately 45% v / v ethanol: pH 4.5; the total acidity, calculated as salicylic acid, was greater than 0.1%. The product is suitable for blonde hair. During the 12 months of storage, the product remained unchanged.

15 Example 9

The procedure described in Example 8 was repeated except that instead of chamomile extract and TILIANA H4308 composition was used in the same sequence and ratio: the herbal extract and the fragrance composition FINUS H4625. The horsetail extract was a 20 green-brown liquid having a density of 0.9160-0.9503 g / ml and an ethanol content of 55% by volume.

The resulting preparation is particularly suitable for all types of hair. It was a clear and translucent liquid with no solid components, yellow-brown in color; The pH, ethanol content and total acidity as well as the stability after 12 months of storage were the same as in the preparation of Example 6.

Example 10

The method described in Example 8 was repeated with comparable results. The only difference was that instead of chamomile extract and TILIANA H4308 composition, nettle leaf extract and fragrance-35 composition LELIA 90368 (product of the same Fabryka Syntetykow Zapacho-Wych Pollena-Aroma) were used in the same section and ratio. The herb extract used was olive green in color, had a density of 0.91 60-0.9503 g / ml and an ethanol content of 55% by volume. The preparation was suitable for all types of hair.

14 103258

Example 11

Generally, cosmetic milks are dispersions of fat substances that act both chemically and mechanically on the skin. In fact, due to the convenient mode of administration and the better fluid-skin interaction, it is very convenient to use liquid creams, more particularly lotions. They can easily penetrate deeper layers of the skin, thus preventing age-related skin changes. Cosmetic milks are mainly used to cleanse dry and sensitive skin. Thus, they must not contain any aggressive volatile oils, while they often contain suitable herbal extracts such as chamomile extract or wheat germ extract. The addition of peat-derived bioactive substances to such compositions further enhances their positive effects. The new recipe is as follows: TTP 0.05 g aloe extract 20.00 g glycerol 3.00 g 20 euserine 2.00 g white paraffin oil 1.00 g triethylamine 1.00 g

Aerosil (R) (colloidal silica) 4.00 g 25

Example 12

Improved refreshing results were obtained when TTP and carefully selected fat carriers were used in the classic nourishing and refreshing formula. TTP 30 is used in an amount of 0.01% to 1.00% by weight in combination with an herb extract (selection depends on the type of skin for which the cream is intended) in an amount of at least 0.05% to 1.00% by weight, an antibacterial 0.05 to 1.00% by weight, a synthetic fragrance compound in an amount of 0.01 to 35 0.05% by weight, and a fatty acid carrier in the form of an aqueous emulsion of 97.00 to 99.50% by weight. -% of total composition. The fatty acid composition must be a good carrier for the active ingredients and be well suited for the skin. It is preferably an emulsion (all amounts by weight) containing, 5 to 10,3258 of 35 to 45 euserines, 8 to 14 petrolatum, 2.5 to 4 olive oils, 6 to 10 glycerol and 35 to 40 water. Preferred herbal extracts are marigold flower extract, chamomile extract, yarrow extract and the like. Preferred recipes are as follows: 5 Euserine 39.00 parts by weight

Vaseline 11.50 parts by weight

Olive oil 3.13 parts by weight

Glycerol 7.80 parts by weight

Water 38.00 parts by weight 10 NIPAGINA A (antimicrobial preparation) 0.40 parts by weight TTP 0.05 parts by weight

Marigold flower extract 0.10 parts by weight

Synthetic fragrance _0.02 parts by weight 15 Total 100,000 parts by weight

Euserine 42.00 parts by weight

Vaseline 8.50 parts by weight

Olive oil 3.08 parts by weight 20 Glycerol 7.90 parts by weight

Water 38.00 parts by weight NIPAGINA A (antimicrobial preparation) 0.40 parts by weight TTP 0.05 parts by weight 25 Chamomile extract 0.02 parts by weight. Synthetic fragrance _0.05 parts by weight

100,000 parts by weight in total

EXAMPLE 13 [0127] The lotion preparation contains TTP as a peat extract in an amount of 0.01 to 1% by weight, herb extracts in an amount of 1 to 30% by weight, X, glyse; roles in an amount of 1-8% by weight of salicylic acid and menthol in an aqueous alcoholic solution. Preferred herbal extracts are: camomile, marigold, yarrow, aloe extract and similar herbal extracts. Glycerol addition is also advantageous due to its effect on skin elasticity. It speeds up the penetration of the preparation into the face as well as its penetration. 16 103258 the beneficial effects of its active peat composition and herbal extracts. Preferred recipes are: TTP 0.10 parts by weight

Camomile Extract 15.00 parts by weight 5 Glycerol 5.00 parts by weight

Menthol 0.10 parts by weight

Salicylic acid 0.10 parts by weight

Ethanol (conc. 95%) 10.00 parts by weight

Fragrance composition 0.30 parts by weight 10 Distilled water ad 100.00 parts by weight

Example 14

A shampoo composition was prepared according to the following recipe: 15 Fuller Herbal Extract 15.00 g 7.50% w / w

Nettle Leaf Extract 20.00 g 10.00% by weight GAMAL SBS-11 (detergent) 3.00 g 15.00% by weight GAMAL NO-3 (detergent) 20.00 g 10.00% by weight

Aseptic 0.40 g 0.20 wt% 20 Ethanol 1.60 g 3.80 wt% BRONOPOL (preservative) 0.04 g 0.02 wt%

Sodium chloride 6.00 g 3.00% by weight

Water 106.96 g 53.48% by weight

A total of 200.00 g of 100.00% by weight of 25 to 92 parts by weight of the above shampoo composition was added with 8 parts by weight of TTP to obtain 100 parts by weight of the shampoo of the invention. Other herbal extracts can be used instead of nettle leaf extract.

30

Example 15

The following shampoo composition was prepared:

Horse chestnut extract 13.00 g

Marigold Flower Extract 22.00 g 35 GAMAL SBS-11 30.00 g GAMAL NO-3 20.00 g

Aseptin 0.40 g

Ethanol 1.60 g BRONOPOL 0.04 g 17 103258

Sodium chloride 6.00 g

Water 106.96 g

A total of 200.00 g of 5 to 95 parts by weight of the above shampoo composition was added with 5 parts by weight of TTP to obtain 100 parts by weight of the shampoo of the invention.

Example 16 Toothpaste contains TTP as a concentrated peat extract in an amount of 0.01-0.10% by weight, essential oils or their compositions or other fruit essences in an amount of 1-10% by weight, herbal extracts in an amount of 0.10-10% by weight and cleansing agents in an amount of 20-35% by weight dispersed in water 15 in an amount of 45-60% by weight and dyes and bleach components in an amount of 1-2% by weight.

Titanium dioxide may be used as a bleaching component; TTP itself can be used as an antibacterial additive; sage leaf, chamomile or marigold flower extracts can be used as beneficial inexpensive herb extracts. The preferred recipe is as follows:

Precipitated calcium carbonate 150.00 g

Magnesium carbonate 60.00 g 25 Glycerol 70.00 g

Herbal extract 5.00 g TTP 0.50 g

Titanium dioxide 10.00 g

Essential oils (mint lemon 30 essences etc.) 5.00 g

Water 400.00 g. Color slightly

Example 17 35 Bath Salt Preparation: A process for obtaining a bioactive product derived from peat within the scope of the invention, in particular when converting a liquid form into a powder form, is a salt. a dewatering step in which sodium chloride is separated as a by-product. In said by-product 95% consists of sodium chloride, the other separated mineral salts are calcium salts, magnesium salts, mainly chlorides and sulphates; salt products also contain some organic peat-derived low-molecule compounds embedded in the crystalline structure of these inorganic salts. These organic compounds are components of TTP and include polysaccharides, amino acids, fulvic acids and the like. The presence of these components in saline byproducts is advantageous if the salt is used as bath-10 salt as they may add other beneficial effects on the standard activity of the bath salt. Therefore, this by-product was tested for chemical and physical properties at the Poznan Balneological Institute in Poland to see if it could be used in cosmetic baths. Since the institute did not find any unwanted salt in the salt, it was approved for cosmetic use. The preferred recipe is as follows:

Salt (NaCl), containing TTP 97.00 g 20 Essential tall oil or essential oils composition 3.00 g

Example 18

The new hair balm contains TTP in an amount of 0.01 to 1 wt.%, Herbal extracts in an amount of 0.01 to 10 wt.%, Anti-electric. , substances in an amount of 3-4% by weight, components which prevent excessive drying of the hair and skin up to 2% by weight, glycerol in an amount of 1-5% by weight, preservatives and stabilizers in an amount of 0.05-0.50% by weight % and 30 water up to 100% by weight.

The present balm contains, as an anticancer agent, an alcoholic solution of trimethylamine and an ammonium chloride salt derived from amines derived from a fatty animal, as a thickening agent, a cosmetic alcohol which also acts as a stabilizer; vegetable oils, which act as co-emulsifiers at the same time as an agent to prevent excessive drying of hair and skin; and glycerol to facilitate spreading and penetration of the balsam, especially its active substances TTP and herbal extracts. Because the acidic medium stops bacterial growth, the balm of the invention contains citric acid or fumaric acid in an amount of 0.1%, as well as a preservative known as BRONOPOL, and fragrance compositions. The preferred recipe is as follows: Trimethylamine and ammonium chloride alcohol solution 3-4% by weight

Cosmetic Alcohols 3-4% by weight TTP 0.01-10% by weight 10 Thickened Herbal Extracts 0.01-10% by weight

Glycerol 1.5% by weight

Vegetable oils up to 2% by weight

Citric acid or fumaric acid 0.1% by weight BRONOPOL 0.1% by weight 15 Fragrance composition 0.3% by weight

Distilled water up to 100.00% by weight

Example 19

Beauty masks are well-known cosmetic ready-to-use 20s that serve many purposes. Medicinal mud has known beneficial effects on the skin and body, it was believed that post-extraction peat obtained in a process for separating bioactive peat-derived compositions from peat can be used for cosmetic applications. The post-extraction peat contains the active bodies, the solution released in the alkaline hydrolysis process due to the extremely high sorption properties of the peat after neutralization. Thus, it was found to be a valuable component of beauty masks. In order to dewater the post-extraction peat ri-30 with several valuable components, natural medicinal sludge and humic acid fractions present in natural peat were added and separated in a process for obtaining the bioactive compositions derived from peat from the alkaline hydrolyzate. Such a composition was tested at the aforementioned balneological institute and found to be suitable for cosmetic use. The preferred recipe is as follows: Post-extraction peat 100.0 g

Natural medicinal mud or peat 20.0 g 20 103258

Humic acid fraction 10.0 g

Magnesium carbonate 10.0 g

Zinc oxide 5.0 g

Citric acid or the like 0.1 g 5 Herb extract or powdered plant material 5.0 g

Distilled water q.s.

The following statements and explanations relate to the biological aspect of the products of the present invention, i. the bioactive features and compatibility of these products, particularly with regard to their use as pharmaceuticals. The following abbreviations are used below: TTP TOLPA (r) Peat Product (Trademark of Torf Corporation, Wroclaw, u. Mydlana 2, Poland) IFN Interferon exists as a ubiquitous cytokine (tissue hormone). IFN genes are present in all cells. Proteins and glycoproteins mainly induce IFN. The substances that stimulate IFN genes for IFN production are called inducers. The IFN induction process is a highly regulated, sophisticated biochemical process, negative and positive regulatory genes controlling IFN production have been identified. Small amounts of IFN butter can. is produced spontaneously without a detectable inducer.

Such IFNs are sometimes referred to as physiological IFNs. "IFN is naturally occurring in three major molecular forms: IFN-alpha (or leukocyte IFN), IFN-β (or fibroblast IFN), and IFN-gamma (immune IFN).

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IFN-alpha and IFN-β are I-IFNs, IFN-gamma is type II-35 IFN

The major biological activities of IFNs are antiviral, antiproliferative (anticancer) and immunomodulatory activities.

21 103258

Various forms of IFN have been commercially produced in natural and combination formulations and are used as drugs for the treatment of neoplastic, viral or various other diseases.

CTL = cytotoxic T lymphocyte NK cells = natural killer cells 11-1, IL-2 = well-known interleukins that stimulate proliferation of T cells and other lymphocytes including B cells; RPMI 1640 = Tissue Culture Medium for Growth of Human and Other Leukocytes (Roswell Park Memorial Institute, Buffalo, abbreviated); FCS = fetal calf serum (for leukocyte testing it must be pre-tested because it may contain mitogenic substances that mimic the function of interleukins); EMCV = encephalomyocarditis virus, a murine picorna virus that is not pathogenic to humans, is often used as an excitation virus in IFN bioassays; A-549 = human adenocarcinoma cell line used in IFN bioassays because of its sensitivity to IFN-alpha, beta and gamma. WHO experts recommend the IFN standard 25 for such use; ; MTT = 3 [4,5-Dimethylthiazol-2-yl] -2,5-diphenyltetrazolium bromide. A reagent used to measure cell death or cell growth in multiple bioassays using ELISA scanners (Hansen et al., J. Imrauno. Meth. 30, 1989, 119, 203-219); L929 = mouse cell line commonly used in mouse. IFN and human or mouse TNF; TNF tumor necrosis factor, a cytokine (a relatively small protein highly susceptible to proteolytic enzymes) 35 produced by monocytes and macrophages (TNF-alpha is also known as a neurotransmitter that causes malignancy in humans and animals); produced by stimulation with LPS (lipopolysaccharides), viruses, bacteria, and many other substances, 22,103,258 highly toxic to many virus-infected and neoplastic cells; can also act as a growth factor for fibroblasts. Associated with inflammatory reactions. The related form TNF-en (lymphotoxin) is mainly produced by T cells and some other cells.

PBL - peripheral blood leukocytes, normal human leukocytes derived from healthy blood donors, isolated from "impermeable membranes" (intermediate between red blood cells and plasma). The reactivity of PBL from individual donors to various cytokine inducers appears to depend on the genetic structure of the donor. Highly reactive and non-reactive have been identified. This also means the reaction of PBL with TTP. 1 5

The variability of a single response to IFN or TNF inducers is more evident with weak inducers than with strong inducers such as viruses. This is because the response to weak inducers is of the "all-or-nothing" type, whereas viruses almost always induce detectable amounts of cytokines.

Inducer Tolerance: Also called hyporeactive state. There is a single dose inducer after administration, for example 20 hours of exposure to the inducer of PBL. after (e.g., virus); cells stop the additional production of IFN. Hyporeactivity usually lasts about seven days. Such reactivity makes the therapeutic application of potent inducers difficult and / or ineffective. Weak IFN inducers do not induce a hyporeactive state or tolerance is low.

: Numerous natural medicines extracted from medicinal knives have immunomodulatory properties. TTP seems to be a-35 one of them. TTP alters many different immune functions in vitro and in vivo. It initiates a balanced immune stimulation with the ability to non-specifically activate all effector pathways (CD4 helper, weakly CD8 suppressor, CTL, NK cell, and activated macrophage) without cytotoxicity to normal tissues.

TTP has a restorative effect on normal wound healing. Low doses of TTP weakly stimulate IL-1 and IL-2 production. High doses can inhibit cytokine synthesis.

Similar tests with TTP were performed using the following procedures:

PBL from healthy blood donors was purified by treatment with ammonium chloride. The culture medium was RPMI 1640 plus 10% FCS. Approximately 8 x 10 6 cells / ml were cultured for 20 hours at 37 ° C, 5% CO 2. Interferon antiviral activity was tested by EMCV cytopathic inhibition in human A549 cells. The MTT method was also used to measure cell death.

TNF activity was measured in 11929 cells. To determine the IFN type, individual IFN samples were treated with various anti-IFN sera for 1 hour. Their antiviral activities were compared to untreated preparations.

Tests show that TTP stimulates the production of endogenous interferons (IFN) and tumor necrosis factor (TNF). The response is dose related.

Seven commercial batches of commercial TTP tested had comparable biological activity as immunostimulant and cytokine inducers.

Significant variation was observed in the TTP response of individual blood donors' leukocytes. 35 PBL from several donors were found to be unresponsive. This may reflect the genetic background. Strong polyclonal antisera such as anti-IFN-alpha (Cantell), anti-IFN-α-ly (Namalwa) from K. Fantes and anti-IFN gamma (Can-24 1 0 3 2 5 8 bogie) were used. to neutralize antiviral activity in PBL supernatants treated with 20 h TTP.

The results of the neutralization tests were found to resemble the fingerprints of a single blood donor. In other words, the proportions of IFN types produced by individuals varied significantly. Separation of PBL into adherent and non-adherent fractions can enhance induced cytokine synthesis.

10

Hyporeactivity (tolerance) for IFN induction with NDV observed 20 h after the initial stimulation with PBL TTP was either minimal or absent.

All seven batches of TTP tested in human PBL for IFN and TNF inducer were found to be active in inducing IFN and / or TNF. The optimum concentration of TTP for IFN induction was 30-100 pg / ml and for TNF induction 100-200 µg / ml. A dose of 200 pg / ml may be subtoxic to 20 PBLs, but TNF synthesis occurs more rapidly than IFN and faster than the development of moderate cytotoxicity.

Conclusion from the foregoing is the experience that the active principle in TTP drugs is the immunoreactive fraction of 25 extracts of peat containing organic, primarily bound sugars, amino acids, uric acids, humic acid substances and mineral salts including trace elements. ld50 in animals is> 2400 mg / kg per os.

No mutagenic, genotoxic, embryo-toxic teratogenic or carcinogenic active substance was detected in TTP. suutta. TTP has no allergenic properties and has no locally irritating activity.

35 Therapeutic indications include chronic and recurrent respiratory tract infections and leg ulcers, vaginal discharge supplementation, and dental peripheral tissue disorders.

25 1 0 3 2 5 8

There are clinical observations that suggest that TTP can be used as an immunomodulator in support of the treatment of several neoplastic diseases. TTP is used orally (5 mg tablets) or topically.

5

Of particular relevance and importance seems to be the fact that TTP treated with PBL for 20 h at 37 ° C does not develop a hypo-reactive state because it retains the ability to produce IFN with NVD (Newcastle Disease Virus), a highly potent IFN inducer, after induction.

Tables 1-24 in the appendix address various biological, for example toxicological, haematological and immunological tests. These tables appear to be self-explanatory and provide the biologist with information regarding TTP compatibility and biological activity.

It is valuable to note that the concentration of the active ingredients in the drugs, as shown in claim 13, is higher than in the cosmetic compositions (shown in claim 14) for the following reasons.

The drugs are prepared in unit doses, with the concentration of active ingredients under strict control; pre-. . for example, in tablets, the concentration is adapted to the size of the tablet containing the effective daily dose (or fraction thereof) of the active ingredient.

The concentration of the active ingredient, for example TTP, in the granules filled with the capsules is preferably only. 5% by weight to achieve sufficient tablet size and allow the encapsulating machine to function properly.

Another reason is that most drugs are administered orally and the active substance is distributed throughout the body. Although it reaches skin cells at very low concentrations, the therapeutic effect is remarkable.

26 103258

In addition, in contrast to drugs, cosmetic compositions are used in unregulated amounts at a locally different cell penetration rate.

5

Some compositions, such as shampoo, which is washed off almost immediately, have short contact times with the body or skin and may contain a relatively more useful component; the others are used several times a day for 10 days and thus should have a lower concentration of active substance.

In addition, in the case of toothpaste in contact with the oral mucosa, penetration of the active agent is easier than transdermal delivery and the concentration of the active agent may be low.

table 1

Acute toxicity. Effect of orally administered TTP on bio-20 chemical parameters of rabbit blood

Genital Dose Testing- Parameters Investigated Half ΊΤΡ Day Creatinine Total Gamma Globulin / kg (mg-%) Protein Line 25 _ (g / 1) (g-%) 0.0 0 1.09 * 0.03 65.0 ± 3.49 0.53 * 0.00 7 1.03 ± 0.02 62.0 ± 0.33 0.56 * 0.01

Female 2.0 0 1.06 * 0.01 58.0 * 1.46 0.50 ± 0.02 30 7 1.14 ± 0.38 57.8 ± 1.54 0.54 * 0.01 5 , 0 0 0.96 * 0.12 70.3 * 3.52 0.55 * 0.00 _7_0.94 * 0.13 60.0 * 2.34 0.57 * 0.00 0.0 0 0 , 80 * 0.11 73.8 * 3.23 0.72 * 0.06 35 7 0.82 * 0.13 75.3 * 1.02 0.79 * 0.03

Male 2.0 0 0.88 * 0.05 64.6 * 2.86 0.62 * 0.10 7 1.24 * 0.34 64.6 * 2.03 0.67 * 0.07 ·! 40 5.0 0 0.85 * 0.03 58.5 * 3.19 0.57 * 0.02 _7_1, 04 * 0.12 57.6 * 4.68 0.56 * 0.01 45 50 27 103258

Table 2

Acute toxicity. Effect of orally administered ΊΊΡίη on the activity of transaminases in rabbit serum 5 Sex Dose Testing Parameters studied ΤΊ5? day Alanine Asparagine g / kg aminotransferase aminotransfer expired _IU_IU_ 0.0 0 8.67 ± 1.20 22.33 ± 2.60 10 7 9.67 * 0.33 28.67 ± 3.75

Naar 2.0 0 10.33 * 1.76 18.33 ± 4.09 7 12.33 * 3.52 20.00 ± 5.03 15 5.0 0 11.33 ± 1.66 22.33 ± 3.52 _7_11.00 * 3.05_20.67 * 4.66 0.0 0 10.33 * 1.76 23.00 ± 3.05 7 12.00 * 2.64 25.67 * 4.17 20 Male 2.0 0 9.00 * 1.00 18.00 ± 1.52 7 9.00 * 0.57 20.66 * 3.52 5.0 0 8.00 * 1.00 18.00 * 1 , 00 _7_11,33 * 1,66_23.00 * 1, 99 25

Table 3

Acute toxicity. Effect of oral administration of TTP on hematological parameters in rabbits 1Q-6 0.0 0 12.4 * 0.44 38.3 * 1.85 7.32 * 0.43 5.21 * 0.20 35 7 11.8 * 0.63 37.7 * 1, 20 6.30 * 0.66 5.11 * 0.11

Female 2.0 0 11.5 * 0.31 36.7 * 0.88 5.69 * 0.58 5.86 * 0.55 7 12.3 * 0.48 38.3 * 0.66 5, 74 * 0.76 5.69 * 0.11 • '40 5.0 0 12.3 * 0.14 40.7 * 2.6 7.35 * 0.99 5.42 * 0.20 _7 11, 6 * 0.23 37.3 * 0.33 8.55 * 1, 79 5.08 * 0.19 0.0 0 12.0 * 0.85 37.3 * 1.20 11.10 * 0.634,53 * 0.23 7 11.09 * 0.6137.5 * 1.85 10.80 * 0.944,54 * 0.19 45 Male 2.0 0 12.8 * 0.26 43.7 * 4.80 9, 94 * 2.00 4.46 * 0.65 7 11.8 * 0.38 34.7 * 2.02 11.06 * 1.08 5.37 * 0.56: 5.0 0 12.7 * 0.63 38.7 * 2.40 9.52 * 1.36 4.59 * 0.16 _7 10.6 * 1.04 4.0 * 2.08 8.87 * 1, 73 5.72 * 0, 66 50 55 28 103258

Table 4

Chronic toxicity. Effect of orally administered TIP on rabbit organ mass 5 TIP dose (mg / kg) 0.0 50 150 0.0 50 150

Gender Female Male

Body Weight as a percentage of total body 10 Lung 0.72 * 0.08 0.61 * 0.17 0.67 * 0,? 0.56 ± 0.05 0.54 * 0.09 0.71 * 0.14

Heart 0.23 * 0.01 0.22 * 0.02 0.25 * 0.03 0.24 ± 0.02 0.23 * 0.01 0.23 * 0.02

Last year 0.057 * 0.01 0.051 * 0.00 0.079 ± 0.02 0.05 * 0.01 0.06 ± 0.01 0.04 * 0.01

Pay 2.85 ± 0.15 3.02 * 0.08 3.22 * 0.37 2.82 * 0.09 2.77 * 0.28 3.12 * 0.06

Kidney 0.59 ± 0.03 0.52 * 0.02 0.50 * 0.01 0.48 * 0.03 0.55 * 0.01 0.49 * 0.03

Adrenal, gland 0.016 * 0.01 0.015 * 0.01 0.020 * 0.01 0.013 * 0.01 0.013 * 0.02 0.012 * 0.0 20 Mura series and uterus 0.58 * 0.04 0.39 * 0.02 0.60 * 0.04

Stones 0.30 * 0.01 0.32 * 0.03 0.36 * 0.02

Table 5

Chronic toxicity. Effect of orally administered 1TP on haematological parameters of rabbits

Genital Dose Testing- Parameters examined half TCP day Hemoglo- Hemato- Leuko- Erythro g / kg Bin Chalk Causes Causes _ (mg -%) _ (%) x 10-3 χ 10 ~ 6 35 Male 0.0 0 12 , 2 * 0.05 38.2 * 1.71 9.33 * 0.79 5.10 * 0.56 7 12.3 * 0.15 40.7 * 0.33 9.39 * 1.18 6 , 34 * 0.23 6 3.0 * 0.35 41, 7 * 1, 30 9.58 * 1.12 7.32 * 0.70 12 13.5 * 0.19 40.7 * 0.67 11.23 * 1.106.30 * 0.46 40 Male 50 0 12.9 * 0.55 35.7 * 1.82 7.52 * 0.78 5.14 * 0.47 3 - 6 13.3 * 0.35 41, 0 * 2.10 8.08 * 1, 17 6.38 * 0.11 12 13.7 * 0.11 39.3 * 0.94 9.36 * 1, 01 6.19 * 0.19 45 Male 150 0 14.20 * 0.21 36.7 * 1, 81 8.19 * 0.16 5.34 * 0.16 3 11.05 * 0.2638.5 * 1.53 8 , 83 * 0.78 5.68 * 0.40 6 12.40 * 0.6337.3 * 1.45 8.18 * 0.06 6.84 * 0.10; _12 13.70 * 0.32 39.0 * 0.68 8.24 * 0.04 6.54 * 0.54

Female 0.0 0 12.6 * 0.27 40.2 * 0.83 7.07 * 1.16 5.63 * 0.17 50 3 12.1 * 0.23 39.8 * 0.72 8 , 16 * 1.01 5.74 * 0.12 6 12.4 * 0.86 42.3 * 1.20 9.78 * 1.69 5.86 * 0.24 12 12.5 * 0.87 37.7 * 2.18 10.90 * 2.16 4.60 * 0.29

Female 50 0 13.2 * 1.20 35.0 * 1.70 7.44 * 0.98 5.47 * 0.50 55 3 - 5 13.0 * 0.55 39.3 * 1.20 7 , 25 * 0.98 5.47 * 0.60 12 12.8 * 0.21 37.7 * 4.33 9.31 * 0.35 6.02 * 0.32 29 103258

Naar 150 O 15.2 ± 1.15 38.3 ± 1.40 8.50 ± 0.54 6.09 ± 0.08 3 13.01 ± 1.10 47.0 ± 2.80 8.10 ± 0.76 5.85 ± 0.57 6 12.5 ± 0.82 41.0 ± 1.15 6.85 ± 0.31 5.61 ± 0.30 _12 13.1 ± 0.70 41.0 ± 1.00 8.15 ± 1.68 5.92 ± 0.13 5

Table 6 Effect of TEP on Phagocytic Activity of Neutrophils in Peripheral Blood in Rabbits 10 Phagocytic Activity

Group n Neutrophils Neutrophils L.H. L.W.

(1000/1 mm3)%, NBT

0360360360 3 6

Control-15 Ki in 1 mL PBS iv. 6 1.9 2,3 2,414,315,117,654,560,659,8 7.53 7.53 7.61 ± 0.9 ± 1.1 ± 0.4 ± 4.6 4.6 ± 3.0 ± 9.4 ± 4.8 ± 6.8 ± 0.77 ± 0.86 ± 0.4 20 ΊΤΡ (test 1) * o * o * o * * * 5 mg / kg 15 2.0 1.2 1.7 18.4 26, 8 27.2 59.2 69.3 74.9 8.75 10.3 9.88 iv ± 0.9 ± 0.7 ± 7.1 ± 7.1 ± 6.7 ± 7.0 ± 6, 7 ± 7.0 ± 7.8 ± 1.18 ± 1.25 ± 1.0

25 TEP

(test 2) * o * o * o * o * o 5 mg / kg δ 2.8 1.6 1.6165.522.1226.563.069.576.5 7.768,52 8.56 iv ± 0.8 ± 0.6 ± 1.0 ± 4 9 ± 7.0 ± 9.1 ± 7.0 ± 7.1 ± 8.9 ± 0.83 ± 1.04 ± 1.0 30 n Number of experimental animals PBS phosphate buffer 0 before administration of TTP 3 3 days ΊΤΡ: η after administration 6 6 days after administration of TTP 35 * Statistically significant variation at p <0.05 relative to value prior to administration of TTP o Statistically significant variation at p <0.05 when compared to control group 40

Table 7 Effect of TTP on rabbit peripheral blood lymphocyte count and percentage of T lymphocytes (E-rosettes) and B-lymphocytes (EAC-rosettes) 45

Group n Lymphocytes E-rosettes% EAC-rosettes% x 10-3 / mnP) 0 6 0 6 0 6

Control 50 1 mL PBS 6 6.6 6.0 19.0 20.1 42.1 42.3 iv. ± 2.0 ± 1.9 ± 5.7 ± 5.5 ± 6.3 ± 2.05 ΊΤΡ (Test 1) * o 5 mg / kg 15 6.6 5.4 19.2 27.1 43, 7 47.7 55 iv ± 1.5 ± 1.4 ± 4.1 ± 3.2 ± 2.6 ± 4.9 30 103258 ΊΤΡ (Test 2) 1ο 5 mg / kg 8 6.0 6.8 17 , 0 24.8 41.6 41.0 iv ± 1.0 ± 0.6 ± 1.9 ± 2.9 ± 1.3 ± 2.1 5 n Number of experimental animals PBS phosphate buffer 0 before administration of TTP 6 6 days after administration of TTP 10 * Statistically significant variation at p <0.05 relative to value prior to administration of TTP o Statistically significant variation at p <0.05 when compared to control group 15

Table 8

Splenocyte count after long-term ΤΓΡ treatment

Control ΊΤΡ 10 mg / kg TTP 50 mg / kg 20 days / day n = 10 n = 10 ΤΓΡ doses of PFC / 106 splenocytes time week 4 day 7 day 4 day 7 day 4 day 7 day 3 367 ± 157 156 ± 41 1120 ± 3281 336 ± 2401 772 ± 2361 151 ± 57 25 5 572 ± 134 138 ± 47 1528 ± 3461 292 ± 621 1239 ± 2801 232 ± 90 7518 ± 45 133 ± 40 699 ± 1361 159 ± 44 722 ± 2581 141 ± 63 9 466 ± 185 175 ± 75 395 ± 94 132 ± 36 412 ± 138 167 ± 62 12 287 ± 18 153 ± 30 287 ± 131 156 ± 39 376 ± 130 167 ± 69 30 1 Statistically significant fluctuation p <0 , 05 compared to the control group

Table 9

Concentration of anti-SRBC (19S + 7S) antibodies after long-term ΤΓΡ treatment 35

Control TTP 10 mg / kg TTP 50 mg / kg / day n = 10 n = 10 40 ΊΤΡ-an- Hemagglutinin, titer -log2 increase time wk 4 day 7 day 4 day 7 day 4 day 7 3.75 ± 1.78 7.12 ± 1.36 8.80 ± 0.971 9.20 ± 1.461 8.20 ± 0.401 6.90 ± 1.37 5 5.82 ± 1.40 6.50 ± 1, 00 7.70 ± 0.841 9.10 ± 2.021 7.70 ± 0.641 9.33 ± 2.901 45 7 3.60 ± 0.48 5.20 ± 0.74 4.40 ± 1.111 4.10 ± 0.831 3.50 ± 0.831 3.55 ± 1.21 9 3.20 ± 1.40 4.66 ± 1.24 3.50 ± 1.28 4.00 ± 0.81 2.90 ± 1.22 6.33 ± 1 , 52 12 4.80 ± 0.74 4.40 ± 1, 01 5.11 ± 1.09 5.50 ± 1.51 5.50 ± 1.51 6.10 ± 1.74 55

Statistically significant variation at p < 0.05 compared to control group-50 at 31,103,258

Table 10

Concentration of Anti-SKBC (7S) Antibodies after Long-Term ΊΤΡ Treatment 5 Control TTP 10 mg / kg TTP 50 mg / kg n = 10 n = 10 ΤΓΡ-an-Hemagglutinin, titer-log2 increase in 10 weeks 4. Day 7 Day 4 Day 7 Day 4 Day 7 3 1.75 ± 0.60 6.37 * 0.85 3.20 ± 0.87 * 5.90 * 1.70 3.80 * 0 , 87 * 4.90 ± 1.74 * 5 2.50 ± 0.70 6.12 * 1.16 2.20 * 0.97 5.70 * 1.10 2.30 * 1.41 5.33 * 0.74 * 7 1.00 * 0.893.80 * 1.090.60 * 0.80 2.50 * 1.11 * 0.55 * 0.83 2.55 * 0.68 * 15 9 0.40 * 0.483.66 * 1.240.30 * 0.64 3.25 * 0.96 0.00 4.33 * 1.24 12 0.80 * 0.973,80 * 0.761.11 * 0.99 4.40 * 1, 35 1.55 * 1.49 4.30 * 1.26 * Statistically significant variation at p <0.05 compared to control group 20

Table 11 IrisTioduLator effect of TTP

Dose n n NPC / 10 ^ E-rosetti-% Hemaggluti- Hemaggluti- 25 TTP splenosvy- Day 4 yoke * yoke * type

mg / kg charge Type 19S + Type 7S

Day 4 7S Day 4 Day 7 Control 30,469 13.6 5.4 8.2 30 ± 111 ± 3.2 ± 1.1 ± 1/3 0.5 10 997x 15.6 6.8X 8, 4 ± 139 ± 1.4 ± 1.0 ± 1.1

35 2.5 10 839x 20.3X 6.9X 9.7X

± 177 ± 4.1 ± 1.0 ± 1.4

5.0 10 746x 23.2X 7.2X 9.2X

: ± 129 ± 4.7 ± 0.6 ± 1.4 40 10.0 10 795x 18.4X 8.3X 10.4 ”x ± 129 ± 4.0 ± 1.8 ± 1.1

25.0 10,560 16.3X 6.1 9.3X

45 ± 145 ± 2.5 ± 1.1 ± 1.1 50.0 10 400 14.7 5.3 6.7: ± 57 ± 4.0 ± 1.0 ± 1.9

50 100.0 10 375x 11.3X 5.1 5.7X

± 67 ± 2.2 ± 0.8 ± 1.6

250.0 10 305X 9.5X 4.8 5.3X

± 67 ± 2.0 ± 1.1 ± 1.7 55 x Statistically significant fluctuation in overall comparison 0.05 (student. T-test) * titer -lcg2 32 103258

Table 12 Daily dose of ΊΤΡ 4 day after 10 days of immunization with SRBC After 5 days of immunization with SRBC ΊΤΡ dose E E-roset-PFC / 10 ^ Hemagglutinin Hemagglutinin

ti-% charges for splenotiter -log2 titer -log2 19S + 7S 7S 19S + 7S 7S

10 Control I 15 13.7 469 5.7 0.8 9.8 8.4 1 x 0.3 ml ± 3.0 ± 125 ± 1.3 ± 1.0 ± 1.3 ± 1.6

Control I 15 15.3 579 6.1 0.6 9.0 8.6 3 x 0.1 ml ± 4.0 ± 143 ± 1.1 ± 0.9 ± 1.7 ± 1.8 15 2.5 mg / kg 15 19.4 * 864 * 7.4 * 1, 1 10.4 * 9.6 * 1 * 0.3 mL ± 3.8 ± 190 ± 1.1 ± 1.1 ± 1.6 ± 1 , 2 2.5 mg / kg 15 19.0 * 769 * 7.1 * 0.5 9.5 9.0 20 3 * 0.1 ml ± 5.0 ± 132 ± 0.8 ± 0.8 ± 1, 0 ± 2.0 10 mg / kg 10 15 19.3 * 795 * 7.0 * 0.8 10.4 * 9.4 * 1 x 0.3 mL ± 4.1 ± 102 ± 0.7 ± 1 , 0 ± 1.6 ± 1.0 25 10 mg / kg 15 17.5 656 7.3 * 1, 3. 8.9 8.8 3 x 0.1 ml ± 3.7 ± 128 ± 1.4 ± 1.2 ± 1.8 ± 1.5 33 103258

Table 13

Effect of orally administered ΤΓΡ: η for 12 weeks on the immunological response of mice immunized with SRBC 5 at 4 days and 10 days

Fig. EFC / 10 6 hemagglutinin FFC / 10 6 hemagglutinin

koa E-roset- Spleno-Titer -log2 E-roset- Spleno-Titer-log2 ti-X charges 19S + / S 7S ti-% charges 19S + 7S 7S

15.2 565 6.0 1.6 14.7 256 8.7 7.9 10 3 ± 3.8 ± 56 ± 0.9 ± 1.3 ± 3.4 ± 67 ± 1.0 ± 1.0 23.7 * 1035 * 7.5 * 2.9 * 23.5 * 406 * 10.4 * 9.7 * ± 7.8 ± 177 ± 0.5 ± 0.2 ± 3.7 ± 88 1, 2 ± 1.0 13.8 422 5.1 0 12.7 209 7.1 6.7 15 5 ± 2.4 ± 63 * ± 0.9 0 ± 2.1 ± 41 ± 1.4 ± 1.6 23 , 0 * 933 * 7.6 * 2.7 * 20.5 * 430 * 11.0 * 11.0 * ± 6.1 ± 248 ± 1.5 ± 1.5 ± 2.7 ± 86 ± 1, 2 ± 1.0 12.8,495 4.6 0.5 14.0 245 8.8 7.9 20 7 ± 2.9 ± 85 * ± 1.1 ± 0.9 ± 5.1 ± 48 ± 1.6 ± 1.6 20.5 * 1120 * 7.9 * 2.1 * 26.9 * 437 * 11.3 * 10.3 * ± 5.3 ± 214 ± 0.6 ± 1.5 ± 7.7 ± 117 ± 1.0 ± 0.4 15.0 531 6.2 0.5 14.1 294 8.7 8.4 25 9 ± 2.1 ± 67 ± 0.8 ± 0.9 ± 2.8 ± 58 ± 1.5 ± 1.5 18.6 * 1049 * 7.7 * 1.9 18.2 * 515 * 11.9 * 10.5 * ± 2.8 ± 184 ± 0.7 ± 1.3 ± 3.4 ± 530 ± 0.8 ± 1.0 15.0 573 7.2 1.4 16.5 266 7.5 7.1 30 12 ± 2.6 ± 143 ± 1.0 ± 1.2 ± 2, 6 ± 58 ± 1.1 ± 0.8 14.1 770 * 7.2 1.7 19.0 294 11.8 * 10.6 * ± 2.7 ± 132 ± 1.4 ± 1.7 ± 4.1 ± 65 ± 0.4 ± 1.0 * Statistically significant variation at p = 0.05 compared to control group-35. Each group consisted of 40 animals.

Table 14

Effect of storage conditions on TTP activity with regard to ability to form E-rosettes 40 E-rosettes-2,

Dose n After 2 months of storage activity Temperature Room temperature Room temperature + 4 ° C in main mode 45 cyt light in the dark

Control 8 13.6 ± 2.4 12.8 ± 2.3 12.8 ± 2.3 12.8 ± 2.3: 0.1 mg / kg 8 15.8 ± 1.7 16.1 ± 3 , 04 15.1 ± 2.9 16.0 ± 5.5 50 1 mg / kg 8 18.0 ± 0.8 * 19.8 ± 4.1 * 15.3 ± 2.8 19.8 ± 2 , 7 * 10 mg / kg 8 20.4 ± 4.2 * 18.0 ± 4.5 * 16.8 ± 2.4 * 16.9 ± 3.5 * * Statistically significant difference p = 0.05: compared to the control group ···· · 55 countries.

34 103258

Table 15 _

Effect of storage conditions on TTP activity considering the number of cells producing type 19S antibodies 5 PFC / 1C) 6 splenocytes

Dose n After 2 months of storage activity Temperature Room temperature Room temperature Room temperature + 4 ° C light in darkness 10 Control 8 571 ± 69 514 ± 128 514 ± 128 514 ± 128 0.1 mg / kg 8 747 ± 144 1039 ± 3261 718 ± 135 1002 ± 2101 1 mg / kg 8 1204 ± 1551 1026 ± 3141 793 ± 1861 1046 ± 3311 15 10 mg / kg 8 1075 ± 2321 1070 ± 2491 869 ± 1601 848 ± 1371 * Statistically significant variation at alpha = 0.5 compared to control group.

20

Table 16

Effect of storage conditions on TTP1 activity, taking into account the effect of forming anti-SRBC-19S + 7S antibodies 25 Dose n After 2 months of storage activity Temperature Poor. heat Room temperature + 4 ° C light in main mode daylight 4. 10. 4. 10. 4. 10. 4. 10.

30 Control 8 5.6 9.8 5.0 9.5 5.0 9.5 5.0 9.6 ± 0.9 ± 0.6 ± 1.1 ± 2.1 ± 1.1 ± 2, 1 ± 1.1 ± 2.1 0.1 mg / kg 8 7.4 11, 4 7.8 10.5 5.4 10.8 6.5 12.6 ± 0.41 ± 1.31 ± 1 , 01 ± 1.71 ± 0.8 ± 0.71 ± 0.71 ± 2.21 35 1 mg / kg 8 6.8 12.2 6.7 13.8 6.2 13.2 6.5 13 , 1 ± 6.31 ± 1.11 ± 0.41 ± 2.11 ± 0.3 ± 0.41 ± 1.31 ± 1.31: 10 mg / kg 8 6.4 11.0 6.2 12 , 0 4.8 10.6 6.0 13.6 40 ± 1.21 ± 1.41 ± 0.6 ± 2.31 ± 1.3 ± 2.41 ± 0.5 ± 1.31

Statistically significant variation at p = 0.05 compared to control group.

35, 103258

Table 17

Effect of storage conditions on ΊΤΡ: η activity, considering the effect of forming anti-SRBC-7S antibodies 5 Dose n After 2 months of storage activity Temperature Poor. heat Room temperature + 4 ° C light in main mode daylight 4. 10. 4. 10. 4. 10. 4. 10.

10 Control 8 0.4 9.2 0.3 8.1 0.3 8.1 8.1 0.3 8.1 ± 0.8 ± 0.7 ± 0.7 ± 1/4 ± 0.7 ± 1, 4 ± 0.7 ± 1.4 0.1 mg / kg 8 0 10.8 0.7 9.8 0.8 10.0 0.7 11.3 ± 1.01 ± 0.9 ± 0.91 ± 0.9 ± 0.61 ± 0.9 ± 1.41 15 1 mg / kg 8 0.8 10.6 0.3 11.5 0.7 10.3 0.7 11.0 ± 0.9 ± 0.41 ± 0.7 ± 1.81 ± 0.9 ± 0.71 ± 0.9 ± 1.01 10 mg / kg 8 0 10.2 0.3 9.8 0.3 10.7 0 , 7 10.3 20 ± 1.31 ± 0.7 ± 1.0 ± 0.7 ± 2.01 ± 0.9 ± 0.91 * Statistically significant variation at p = 0.05 compared to control group.

25 Table 18

Hematological parameters of healthy volunteers before and after 2 weeks of TTP at 1 mg / day

Parameter Placebo n-5 TEP n-5 30 0 14 days 0 14 days

Erythrocytes x 1θ6 / μ1 4.44 ± 0.46 4.32 ± 0.42 4.32 ± 0.32 4.18 ± 0.32

Hemoglobin 12.4 ± 2.5 11.9 ± 2.8 12.8 ± 0.8 11.8 ± 0.6 35

Hematocrit% 38.1 ± 6.7 38 ± 6.9 38.7 ± 2.1 38.1 ± 1.6: Leukocytes 40 x 103 / μ1 5.25 ± 0.59 5.27 ± 0.8 5 , 23 ± 0.6 4.86 ± 0.6

Neutrophils% 52.6 ± 12 57.6 ± 11 54.2 ± 2.5 55 ± 3.6 45 Lymphocytes% 26 ± 7.5 30.4 ± 8.3 34 ± 4.1 32.4 ± 5, 3

Platelets x 103 / μ1 220 ± 40.4 214 ± 35.8 221 ± 38 314 ± 39.6 50

Blood flow mm / 1h 7.6 ± 9 5.6 ± 1.9 7.8 ± 2.7 6.8 ± 3.1 36 103258

Table 19

Biochemical parameters in healthy volunteers before and after 2 weeks of ΊΤΡ administration at 1 mg / day 5 Parameter Placebo n = 5 ΤΓΡ n = 5 0 14 days 0 14 days

Total Protein g / 1 72.8 ± 1.76 68.42 ± 3.2 73.8 ± 1.92 70 ± 5.5 10 Albumin,% 63.2 ± 2.5 63.8 ± 2.3 63.6 ± 1.8 64.4 ± 2.1

Globulin, alpha% 3.1 ± 0.4 3 ± 065.9 2.8 ± 0.5 2.8 ± 0.45 15 Glculin, alpha2% 5.95 ± 1.97 6.8 ± 1.5 6 , 93 ± 0.9 06.9 ± 0.9

Globulin, guideline% 12.2 ± 1.5 12.1 ± 1.3 13.7 ± 0.9 12.3 ± 1 20

IgG g / l 10.3 ± 1.1 10.2 ± 1 11.7 ± 1.6 11.1 ± 1.4

IgA g / l 1.8 ± 0.6 1.9 ± 0.5 1.8 ± 0.5 1.9 ± 0.5 25 IgM g / l 1.4 ± 0.3 1.4 ± 0, 2 1.1 ± 0.2 1.9 ± 0.5

Complement C3 g / l 1.1 ± 0.1 1.1 ± 0.1 1.1 ± 0.1 0.2 ± 0.2 30 Complement C4 g / 1 0.2 ± 0.5 9.3 ± 0.04 0.2 ± 0.0 0.2 ± 0.03

Alanine U / L 34.2 ± 0.05 0.3 ± 0.04 30.2 ± 8.2 31.2 ± 3.2 35 Asparagine U / L 32.8 ± 4.4 40.2 ± 6, 6 30.4 ± 7.1 39.6 ± 6.6 1 37 103258

Table 20

Comparison of the incidence of acute respiratory infections in the last quarters of 1989 and 1990 with respect to two patient groups treated with TIP and placebo, respectively, October 5 5 aikana last quarter 1990 n 4 quarter 4 th statistical 1989 (ΤΊΡ ) significance 10 mean infections "p" mean average per patient per patient

Colds 20 4.0 ± 1.3 1.0 ± 0.5 <0.01 15

Sore throat 20 3.3 ± 1.3 0.8 ± 0.6 <0.01

Fibrous sputum 20 2.1 ± 2.2 0.3 ± 0.4 <0.01

Cough 20 1.4 ± 1.1 0.3 ± 0.6 <0.01

Bronchial leaflets. 20 0.3 ± 0.5 0

Pneumonia 20 0.2 ± 0.9 0

Placebo 30 given in the last quarter 1990 n 4 quarter 4th quarter statistical 1989 (not TIP) 1990 (TTP) significance of infections infections "p" mean 35 per patient per patient

Colds 20 3.9 ± 1.4 3.1 ± 1.5> 0.05: Sore 40 throat 20 2.3 ± 2.0 2.6 ± 1.7> 0.05

Fever gout 20 2.3 ± 1.8 1.7 ± 2.1> 0.05 45 Cough 20 1.5 ± 1.5 1.9 ± 2.0> 0.05

Bronchi; kentulehd. 20 0.1 ± 0.3 0.1 ± 0.3 50 In pneumonia 20 0.1 ± 0.3 0.1 ± 0.3 38 103258

Table 21

Effect of intradermal injection of bacterial antigens or PHA (phytohemagglutinin) in patients treated with TTP: 11S

5 Series Patients Tuberculosis-Streptolysis-Staphylckts / PHA NaCl No. (Initial) LineRT23 Lysine 0 Anatoxin 0.9% pre 3 pre 3 pre 3 pre 3 pre 3 1 BT + + ± begin - t --- 2 PE - + + ± ± - - - - - 10 3 MH ++++++++ ± - - _ 4 WH - - + - - - - - - 5 WH - ± __ ± _ ____ 6 ZN ++++ - - - - - 7 PJ - +++++ - + --- 15 8 SM + - +++ - ± + --- pre pretreatment 3 after 3 weeks 20 Table 22

Effect of intradermal injection of bacterial antigens or PHA in untreated (placebo) patients

Series Patients Tuberculosis-Streptolysis-Staphylocytes / PHA NaCl # 25 (Initial) LineRT23 Lysine 0 Anatoxin 0.9% pre 3 pre 3 pre 3 pre 3 pre 3 1 MM - - + + ± - - - - - 2 OT ++ - - ± ± ____ 3 OH ++ +++ ++ ++ ± - --- 30 4 WJ --- ± - - ____ 5 KC - - + + ± + - - - - 6 MJ ++++++ + - + - - - 7 BW - - +++ - + --- 8 KT - - ++ + + - - - 35 pre-treatment 3 after 3 weeks 39 103258

Table 23

Immunoglobulins in the serum of patients with TTP-treated tibial ulcer 5n Test group mg mg% pre-treatment after treatment 1 IgG 1732.87 * 207.52 7 IgG 1732.87 * 207.52 8 A 2 IgA 304.25 * 47.50 8 IgA 283.87 * 44.10 3 IgM 202.87 * 71.12 9 IgM 189.75 * 75.15 10 4 IgG 1924.37 * 246.25 10 IgG 1972.87 * 239.33 8 B 5 IgA 296.50 * 59.84 11 IgA 285.50 * 56.76 6 IgM 242.12 * 56.95 12 IgM 232.87 * 65.45 15 1 vs 7 p> 0.05 4 vs 10 p > 0.05 2 vs 8p> 0.05 5 vs 11p> 0.05 3 vs 9p> 0.05 6 vs 12p> 0.05 1 vs 4p> 0.05 7 vs 10p> 0 , 05 20 2 vs 5p> 0.05 8 vs 11p> 0.05 3 vs 6p> 0.05 9 vs 12p> 0.05

Claims (10)

103258 40 A process for the preparation of bioactive products derived from peat from a highly concentrated solution of inorganic salts, in particular sodium chloride containing bioactive ingredients derived from peat, characterized in that said solution is diluted with desalted water and subjected to reverse osmosis to remove salts from the solution. salts 10 are removed and that the resulting solution is concentrated and clarified and optionally sterilized and / or spray dried in at least one step. Process according to Claim 1, characterized in that said concentrated solution is obtained by primary and secondary alkaline hydrolysis of air-dried crude peat material, followed by acidification of the hydrolyzate, separation of insoluble parts and elimination of gravel substances by extraction with organic solvents. and removing any remaining organic solvents from the after-leach water phase, after which the solution is filtered through a sintered ceramic filter under reduced pressure. 3. A process according to claim 1 or 2, characterized in that said solution is diluted with water, preferably distilled, in water volumes several times, preferably 5-8 times, the volume of said solution. Method according to any one of the preceding claims, characterized in that the solution is sterilized by means of a membrane filter, e.g. a Millipore® filter. Method according to any one of the preceding claims 35, characterized in that the solution is spray dried at an inlet temperature of about 180 ° C and an outflow temperature of about 90 ° C. 4i 103258 Process according to any one of the preceding claims, characterized in that the bioactive product derived from peat is kept in the form of a concentrated solution. Process according to any one of claims 1 to 5, characterized in that said product is kept in the form of a powder. A process for the preparation of a pharmaceutical formulation containing a peat-derived bioactive product according to any one of claims 1 to 7 in the form of a gel characterized by combining a sterile solution of a pre-powdered bioactive product of peat, a sterile compound, a flavoring compound, preferably selected from the group consisting of menthol, thymol, mint, lemon, and eucalyptus, and a gelling composition such as sterile glycerol with colloidal silica, to convert the liquid composition into a gel form at a 6. 9. A process for the preparation of a pharmaceutical formulation containing a peat-derived bioactive product according to any one of claims 1 to 7 in the form of an ointment, characterized in that a sterile solution of the peat-derived bioactive product is gradually incorporated into a sterile herb extract so that the resulting mixture forming a compound such as colloidal silicon dioxide and triturating the gel thus obtained with a pre-sterilized mixture of fat components such as euserine and petroleum jelly, preferably in a weight ratio of liquid components to silica of about 30: 2 and a gel fat composition of 32: 68-34: 66. A process for preparing a pharmaceutical composition, wherein the peat-derived bioactive product produced by the process of any one of claims 1 to 7 is admixed with a pharmaceutically acceptable carrier material preferably in a weight ratio of about 1: 5 to 1:25 and most preferably 1: 9 to 1: 19. 5