FI68174B - FOERFARANDE FOER FRAMSTAELLNING AV CYKLODEXTRIN-KAMOMILL-INKLUSIONSKOMPLEX FOER PHARMACEUTISKT BRUK - Google Patents

Description

advertisement publication / n17i JÄMÄ 8 11 UTLÄGGNINGSSKRIFT OO I / 4 C Patent patent 12 CC 1935

Patent Meddelat '(51) Kv.lk.4 / lnt.CI.4 A 61 K 35/78, 31/715 ^ Q | y | | FINLAND (11) Patent application - Patentansökning 793534 (22) HakemlspSivi - Ansökningidag 08.11 .79 (H) (23) Alkupiivi— Giitighetsdag 08.11 .79 (41) Tullut lulklseksl - Blivit offentlig 21 .05-80

National Board of Patents and Registration / ^ \ Date of publication of Nihtäväkilpinon | -

Patent and registration authorities V 'Ansökan utlagd och utl.skriften publicerad 30.04.85 (32) (33) (31) Pyydetty etuoikeus — Begärd priority 20.11.78

Hungary-Hungary (HU) CI — 1867 (71) CHINOIN Pharmaceuticals Chemical Voltage Products Gyära RT, To u. 1-5, H-1045 Budapest, Hungary-Hungary (HU) (72) Jözsef Szejtli, Budapest, Lajos Szente, Budapest, György Körmöczy, Budapest, P £ ter Tötönyi, Budapest, Ägoston Dävid, Budapest,

Tibor Zilahy, Budapest, Andräs Kelemen, Budapest, Jänos Härshegyi, Budapest, Hungary-Hungary (HU) (74) Oy Koister Ab (54) Method for the preparation of pharmaceutical cyclodextrin-kamomi11 aink lysis complexes - Förfarande för framstäl1-Ning av cyclodextrin This invention relates to a process for the preparation of pharmaceutically acceptable cyclodextrin-chamomile inclusion complexes.

Chamomile (Flores chamomillae) and the hogweed (Achillea Collina) and Roman chamomile (Anthemis nobilis) widely used in therapy have been known for a long time and have been used in therapy.

Plant flowers are used internally as a decoction for intestinal inflammation, gastritis, and cystitis, and they are effective as an external bath in conjunctivitis, sinusitis, and otitis media. Namely, the camatsulene contained in the essential oil of chamomile releases histamine, activates the reticuloendothelial system and accelerates the healing process of the turbid-like persistent inflammatory process.

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The use of the chamomile substance can take place in two ways: 1) chamomile is made into either a kettle (f-chamomile tea), which contains flavonoids, coumarins and chamomile-specific fragrances and aromas, but which contains only a small proportion of the essential oil-terpene components, 2) made from chamomile oil. the oil is not used as such, but is used to make ointments, ointments, powders, ie generally anti-inflammatory preparations, the stability of the oil is poor, the preparations containing the oil quickly lose their shelf life, and the oil does not have the well-known aroma of chamomile.

Important active ingredients of chamomile oil: 1-Bizabolol camatsulene dicycloether

Bisabolol oxide I

Bisabolol oxide II

Phamezen

herniarin

A further disadvantage of the use of flowers is that the concentration of the active ingredient decreases continuously during storage and the use of the herb is associated with significant microbiological and also parasitic infectious phenomena. Although cinnamon oil extracted from the flower has no effect on infections or contamination, its stability is even worse. The active substance content of preparations containing blue oil can only be guaranteed for a short time. Thus, stabilization of the active ingredient has not been resolved in any commercial preparation.

The composition of commercial chamomile preparations (in liquid or tablet form) is quite different from that of natural chamomile, containing only a small proportion of the original active ingredients and mainly disintegration products. The products resemble chamomile only for a short time, after which substantial differences can be identified immediately by chemical analysis.

The commercial liquid product was analyzed chromatographically and six months after the desired sale, the preparation could be shown to contain a total of two non-degradable chamomile oil components. The second component was camatsulene, which was only 3 68174 15-20% of the camatsulene content of chamomile oil. However, it is also likely that the substance considered as camatsulene is only azulene (artificially produced azulene). Of the chamomile drops, only small amounts of 1-Bizabolo and two other - probably - degradation products were detected. The product smells slightly burnt and is a brown alcoholic liquid.

The presence of a single flavone compound was detectable, but no chamomile flavon could be identified.

The "Instant Tea" made from steamed dried chamomile tea was also analyzed. Based on chromatographic analysis, the tea does not contain any of the volatile oil components listed above. A special reagent for terpenes could be used to detect the presence of an unidentified degradation product. The product contains a large number of flavones, which are probably different glycosides of the Apigenii-ni flavone.

As noted above, chamomile active ingredients that are chemically unstable and degradable cannot normally be stored and are difficult to homogenize due to their consistency (oil). It has now been found that the active ingredients of chamomile can be stabilized by forming a chamomile inclusion complex of cyclodextrin.

The invention relates to a process for the preparation of new pharmaceutically usable cyclodextrin-chamomile inclusion complexes. The process is characterized in that the α, β or β-cyclodextrin or a mixture thereof is reacted as a saturated aqueous solution with a 10-20% by weight ethyl alcohol solution of chamomile extract with vigorous stirring and, if desired, the cyclodextrin-chamomile complex formed from the cyclodextrin complex is isolated.

The consistency of the complex prepared by the process of the invention is such that it is easy to administer and is more stable than the original substance despite the dispersed state. In addition, it can be moistened with water, which greatly improves its absorption and bioavailability. The improved stability can be explained by the formation of a molecular inclusion complex, i. 1 mole of active ingredient is surrounded by 1 mole of cyclodextrin, whereby the individual components of the chamomile extract are unable to react with each other and a redox system state is formed, which impairs the stability.

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Cyclodextrins are closed rings formed by glycopyranos, the steric form of which can be characterized as a combination of a hydrophilic surface and a hydrophobic closed interior. Thus, when these cyclodextrins are reacted in aqueous solution with hydrophobic molecules, so-called inclusion complexes are formed. The drug molecules enclosed in the inclusion complex are remarkably well protected from external influences (such as oxygen, heat, light, etc.).

The inclusion complex comprises an extract prepared from the plants Flores chamomillae and / or Anthemis nobilis and / or Achillae Collina, and, if desired, flavorings and fragrances extracted from the chamomile herb.

An extract prepared from the plants Flores chamomillae, Achillea Collina and Anthemis nobilis can be used in carrying out the process according to the invention. The extract - so-called cyanobacterial oil - is made from plant material in a known manner. The formation of the complex is carried out at 15 to 80 ° C, preferably at 50 ° C, after which the reaction mixture is cooled with efficient stirring.

The cyclodextrin-chamomile-in-inclusion complexes obtained according to the invention are stable, powdery products which dissociate in water and thus have chamomile activity.

The inclusion complexes can be formulated into pharmaceutical preparations in such a way that there is no risk of degradation of the active ingredient. In preparing the preparations, the usual carriers, binders, lubricants, flavoring agents and, if desired, other substances having a pharmaceutical effect can be used.

It is also possible to prepare combined preparations containing a cyclodextrin complex with chamomile blue oil and a cyclodextrin complex with flavoring and / or odorants in the desired proportions.

The use of the blue oil inclusion complex as a pharmaceutical preparation and a pharmaceutical intermediate, e.g. in powders and ointments, is particularly advantageous, since hitherto only pharmacologically inactive fragrances and flavorings have been used for this purpose as aqueous extracts.

The use of the complexes prepared according to the invention offers the following advantages: 1. The complexes are prepared from naturally occurring active ingredients.

2. All pharmacologically important components of the active substance remain unchanged.

3. The product is stable to heat, light, oxygen and metal ions.

4. The product is a crystalline tabletable powder.

5. Easily dosable pharmaceutical preparations can be prepared which have a stable active ingredient content.

In the following, the properties of the complexes obtained by the method according to the invention are examined in more detail.

Aqueous solutions of the complexes are always slightly opalescent. The complex is stable at room temperature and can be stored without decomposing for at least five years.

The oxidation stability of the product of Example 1 below is shown in Table I.

Table I

Time Oxygen consumption ul / mg hours Chamomile oil fi- CD-chamomile complex 70 6 2 80 14 4 120 18 5 160 23 7 200 30 7 240 58 7 280 80 7 320 100 7

The oxygen consumption of the complex can be ignored compared to the oxygen consumption of pure chamomile oil.

When administered orally, neither chamomile oil nor cyclodextrin is toxic: The two components do not form a hard bond and the complex dissociates under physiological conditions.

Demonstration of activity by studying the effect of the reticuloendothelial system

The phagocyte function of the reticuloendothelial system is caused by histamine as a chemical activator. Under the influence of histamine, 6 68174 outer capillary endothelial cells are converted in a short time into phagocytes, which accumulate in the pen, injected like liver, spleen or bone marrow RES cells.

Because chamomile also releases histamine and activates RES, the effectiveness of pure chamomile oil and inclusion complex can be compared by monitoring the elimination of a pen injected into a blood vessel.

Male CFLP mice weighing 25 (-2) g (CFLP means international strain according to reported genetic information) were used as experimental animals. The methodology corresponds in practice to the methodology of Jansc6 (Jansc6, M .: Orvosok Lapja, 1947, III, 28, 1015). 0.5 ml of 10% felt-tip pen (body weight 25 g) in physiological saline was injected into the tail vein. To this solution was added 1% gelatin to stabilize the colloidal system. A suspension with carbon particles only 0.2-1 yU in diameter was used because the solution containing larger particles clogs the pulmonary veins and the animals die from blockages. Prior to injection of the pen into the stomach of mice, 0.5 ml of physiological saline was injected twice. One hour before the injection of the pen, the animals were orally administered 5 mg of chamomile oil and 5 ml of the chamomile oil-containing product of Example 1. The elimination of the pen was monitored photometrically.

After 5 minutes of the pen treatment by intravenous injection, every 5 minutes, 10 ml of blood was aspirated with a measuring pipette, the blood was homogenized in weakly alkaline water, and then the light absorption of the solution was determined photometrically.

Elimination of pen from the blood of control and treated animals was summarized in Table II.

Toxicity studies of the chamomile-cyclodextrin-closure complex showed that the complex was non-toxic.

10 female mice (body weight 20-25 g) received oral chamomile complex as a suspension at a dose of 3000 mg / kg body weight equivalent to 234 mg active ingredient / kg body weight), or 10 male mice (body weight 20-25 g) received the same conditions complex at a dose of 3000 mg / kg body weight (equivalent to 264 mg / kg body weight) and no toxicity was observed. 10 female rats and 10 male rats (body weight: 150-200 g) received 5,000 mg / kg body weight of chamomile complex (= 430 mg active ingredient / kg body weight): no toxicity was observed in this case either.

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Table II

Elimination of pen from the blood

Pen administration increased blood extinction by an average of 25%, the values in the table show a decrease in groups of 8 animals (on average)

Time since giving the pen .jqq ^ \ _

Eo control Chamomile oil Product of Example 1 between 10-20 minutes X 2.63 7.78 6.75 between 20-30 minutes 3.43 3.30 5.60 between 30-40 minutes 1.69 5.47 4, 67 between 10-40 minutes total 8.15 16.55 17.12 x = The exact value of the extinction for time 0 cannot be determined.

Efc = extinction measured after t pen injection at time t.

E = Extinction measured before pen injection. o "Sensory quality determination of Kompletf camomile

The product of Example 1 contains all the pharmacologically important components of chamomile, but not the components which give the chamomile its characteristic odor. In most cases, this is also not necessary, but in some cases, odor may also be desirable. The product of Example 2 contains these fragrances, so "Komplett" chamomile tea can be prepared by combining the two products.

Accordingly, the following models are compared: 4.5 g of the product of Example 2 (chamomile-aroma complex) + 0.5 g of the product of Example 1 (chamomile oil complex) per liter of 68174 water and 2.2 g of chamomile flowers boiled per liter of water.

The study was performed using a control sample. The models were chamomile tea and tea from the cyclodextrin inclusion complex. The models were warmed to 20 ° C or 40 ° C, and to eliminate the subjective effect of the different color of the models, the models were first placed in paper-covered containers.

The sensory study was performed by 11 observers at 20 ° C and 40 ° C.

Identity was correctly established by 10 individuals, of which 3 individuals rated the chamomile flower tea as better and 7 individuals had found the cyclodextrin-inclusion complex mixture to be better.

Accordingly, the significant value of the discrimination test is 99.9%. The significant value of the quality test is 99.0 S.

Observers who gave the correct answer rated the tea obtained by dissolving the cyclodextrin inclusion complex with a significant value of 99.9% better.

1 g of the product of Example 1 corresponds to 8.40 g of air-dried herb. The product can be used in the veterinary field - in various cases of intestinal inflammation in pigs, cattle, horses, dogs and cats - as follows.

a) In cases where gastric or intestinal inflammation due to a disturbance of posture nutrient-, administered to an animal after fasting 50-150 mg pack-dimethylsulfide complex / kg body weight, or an equivalent dose of a tablet containing 3-5 times for 2-3 days until the diarrhea has stopped. The product can be combined mainly in therapy with ordinary amounts of activated carbon. Until complete recovery, the animal is given a body temperature aqueous solution prepared from the chamomile complex of Example 1 instead of drinking water.

b) In cases where various severe gastrointestinal inflammations are caused by pathogens (coli, etc.), the chamomile complex may be combined with other antibacterial drugs, e.g. oxolinic acid, after laxative treatment. This combination can be administered either in drinking water or mixed with feed until the animal recovers. Even in these cases, the animals are also given an aqueous solution of the chamomile-cyclodextrin complex instead of drinking water until the animals recover.

(c) In the case of external inflammation of various domestic animals, a chamomile-cyclodextrin-containing powder or granulate placed between gauze dressings is placed on the inflamed skin, the gauze dressing being briefly immersed in water before application.

d) A solution or suspension prepared from the product of Example 1 is used in conjunction with conjunctivitis or oral cavity inflammation, in case of anal inflammation or for rinsing the uterus.

e) A few days after weaning, piglets are 30-90% likely to have non-specific, ie non-infectious, stomach. The frequency and intensity of the abdomen can be reduced, or the number of days required to treat the abdomen can also be reduced if chamomile-ε-cyclodextrin prepared according to Example 1 is used in the feed. At the same time, an increase in feed intake or an improvement in feed utilization can be observed.

To 3.42 kg of the camomile-oil-β-cyclodextrin complex prepared according to Example 1 was added 50 kg of flour as a filling and mixed into a homogeneous mixture. The premix was diluted with the feed to 200 times its volume, thus producing 10,000 kg of weaned pig feed with 0.5 mg (daily feed intake of weaned piglets of weaning age) of 15 mg of chamomile oil. The experiments were performed on two different pig farms. On Farm I, tetrahydride pigs were weaned at 28 days of age and fed dry fodder according to the recipe of the Hungarian Phylaxia factory by the self-feeding method. On Farm II, similar pigs of the same age were fed dry feed from a trough according to the Centra Soya recipe. The number of animals and the experimental results are obtained from the following tables.

The tables show that the average weight gain per day was substantially better in the chamomile complex-treated group than in the control group, while the amount of feed required for one kilogram of weight gain was lower at the same time.

The daily weight gain of the animals was 25-30 grams higher and at the same time about 100-125 g less feed was needed to gain one kilogram of weight.

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Farm I

Dead, weight gain and feed utilization parameter

Chamomile Control-treated group group

Age (in days)

At the beginning of the experiment 29.56 + 4.60 25.50 + 2.14

At the end of the experiment 49.56 + 4.60 45.50 +2.14

Number of animals (pcs

At the beginning of the experiment 309 2Qg

At the end of the experiment, 299,293

Dead (10 pieces (3.24%) 13 (4.25%)

Intestinal inflammation 8 (2.59%) 12 (3.92%)

Other reason 2 (0.65%) 1 (0.33%)

Average weight (kg)

At the beginning of the experiment 6.29 + 1.12 5.58 + 0.71

At the end of the experiment 12.88 t 2.12 11.47 +1.17

Average weight gain (kg) 6.59 5.89

Average weight gain per day (g) 330 295 Total amount of feed used (kg) 4350 4050

Average feed consumption per day (g) 715 676 Amount of feed required for 1 kg of weight gain (kg) 2,171 2,296 11 68174

Farm II

Dead, weight gain jr. feed utilization parameter

Chamomile Control-treated group group

Age (in days)

At the beginning of the experiment 33.17 + 3.54 29.68 + 2.84

At the end of the experiment 60.17, + 3.54 56.68 + _ 2.84

Number of animals (pcs)

At the beginning of the experiment 234 211

At the end of the experiment 225,206

Dead (pcs) 9 (3.85 SK) 5 (2.37%)

Intestinal inflammation 6 (2.56%) 3 (1, 42%)

Other reason 3 (1.28%) 2 (0.95%)

Average weight (kg)

At the beginning of the experiment 8.93 + 1.35 7.95 + 1.14

At the end of the experiment 17.20 + 2.23 15.54 _ + 2.10

Average weight gain (kg) 8.27 7.59

Average weight gain per day (g) 306 291 Total amount of feed used (kg) 4850 4205

Average feed consumption per day (g) 783 747 Amount of feed required for 1 kg of weight gain (kg) 2,556 2,658

The following is an example of a suitable composition containing a complex prepared according to the invention.

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Teetabletit

Composition: Chamomile extract-f-CD 20.0 g

Atsulenone 100% men fb -CD 20.0 g

Ascorbic acid 44.0 g

Sodium bicarbonate 15.0 g

Glycerin monostearate 1.0 g

Granules are prepared from ascorbic acid and sodium hydrogencarbonate with glycerin monostearate according to the general guidelines of pharmaceutical technology. The chamomile and azulenol-CD occlusion complexes are thoroughly ground to a dust so that the mixture can be screened without residue through 0.16 mm sieve holes (Ph.Hg.VI sieve classification). Chamomile and azulenol-β-CD occlusion complexes are mixed with ascorbic acid and sodium bicarbonate granules, followed by compression of 0.5 gram tablets.

Tea

Component ratio in the preparation of tea: 1.8 g of chamomile herb extract - / ^ - CD complex and 0.2 g of chamomile oil /? -CD complex is dissolved in 100 ml of hot water.

The following examples illustrate the method of the invention.

Example 1 10.0 g of β-cyclodextrin (moisture content: 14.46%) are dissolved in 180 ml of a 30% strength by volume aqueous ethanol solution at 50 [deg.] C. with constant stirring. A solution of 1 g of chamomile oil (blue oil) in 10-fold diluted ethanol is added dropwise to the β-cyclodextrin solution. 3-4 minutes after the addition, the system becomes cloudy and a crystalline adduct begins to separate. After 4.5-5 hours, the mixture is then slowly cooled to room temperature. Stirring is stopped and the complex is kept in the refrigerator for 16 hours. The mixture is filtered through a glass filter and dried over phosphorus pentoxide at room temperature. 10.06 g of crystalline chamomile oil-cyclodextrin complex are obtained.

The active ingredient content of the complex is 8.12% and the yield based on chamomile oil is 75.04%. The complex formation efficiency can be improved by using a stirrer with a higher speed.

The empirical formula and molecular weight can only be reported as β-cyclodextrin (CgH 2 MgCD: 1135.0. The average molecular weight of the sesquiterpene derivatives of chamomile oil is about 230, the average molecular weight of the complex is about 760 considering the water content of about 1460.

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The structure of the product can only be reported for cyclodextrin, all other active ingredients of the chamomile active ingredient are not yet known.

The product prepared according to Example 1 is a light blue, microcrystalline powder which smells only slightly of chamomile. The product has no melting point, it begins to decompose at about 200 ° C.

This temperature is a typical decomposition temperature of γ-cyclodextrin.

Solubility: in distilled water; 0.030-0.035 g / 100 ml, in 96% ethanol: 0.030 g / 100 ml; only oil is soluble in benzene and chloroform, β-cyclodextrin is practically insoluble in these solvents (values reported are based on 25 ° C).

Example 2

Preparation of the chamomile-aroma-β-cyclodextrin closure complex 10 g of β-cyclodextrin was dissolved in 30% v / v aqueous ethanol at 50 ° C, followed by the addition of 2.5 g of chamomile aroma (chamomile extract) obtained from 30 g. chamomile flower dissolved in 10 ml of ethanol. The system clouds after 10-12 minutes of the addition, after which the heating is stopped and the reaction mixture is cooled to room temperature over 4 hours with constant stirring. The mixture is kept in a refrigerator for 16 hours, filtered through a glass filter and dried over phosphorus pentoxide.

10.8 g of an off-white crystalline product are obtained, containing about 12% of active ingredient (% by weight).

Claims (5)

681 74 14 A process for the preparation of novel stable cyclodextrin-chamomile inclusion complexes for pharmaceutical use, characterized in that ° (-, or? and, if desired, the cyclodextrin-chamomile inclusion complex formed is isolated from the reaction mixture. Process according to Claim 1, characterized in that a chamomile extract obtained by drying the chamomile flowers and extracting the obtained material for 6 to 8 hours at 40 to 60 ° C with petroleum ether or a mixture of petroleum ether and methanol in a volume ratio of 9 to 6: 1 to 4 is used, which after which the solvent is evaporated. Process according to Claim 1, characterized in that cinnamon oil (essential oil) obtained by drying the chamomile flowers and the substance obtained by steam distillation for several hours is used as the chamomile extract. Process according to Claim 2 or 3, characterized in that chamomile extract prepared from the plants Flores chamomillae and / or Anthemis nobilis and / or Achillea Collina is used. Process according to Claim 1, characterized in that the complexation is carried out at 15 to 80 ° C, preferably at 50 ° C, and the reaction mixture is cooled slowly with vigorous stirring.