CZ20003741A3 - Retarded tablets prepared from linear polysaccharides insoluble in water and their use - Google Patents

Abstract

The solution is a retarded tablet which as retarding material it comprises linear or water-insoluble linear a polysaccharide, preferably a poly- (1,4-alpha-Dglucan) polysaccharide in the form of microparticles and is adapted to be controlled release of the active ingredient.

Description

Technical field

The present invention relates to retarded tablets of linear water-insoluble polysaccharides, a process for their preparation and their use, in particular for the controlled release of active substances.

BACKGROUND OF THE INVENTION

In modern pharmaceutical technologies, the formulations of active ingredient carriers (excipients) are of importance, the dosage form of which specifically affects bio-distribution, bioavailability, bio-compatibility and resorption. In addition, the active compound carriers must have good mechanical properties, such as sufficient hardness and resistance to tension and tension.

Although some compounds may already be compressed into compact, stable masses (e.g., sucrose or lactose), additional ingredients - also excipients for the manufacture of tablets - such as binders, fillers, glidants and additives are needed. Typical dry stability binders used herein are calcium phosphates, microcrystalline cellulose, (e.g., Avicel®, PH102, especially Celsphere), polyvinylpyrrolidones (e.g. Kollidon®, Luviskol VA 64 ^, Plasdone®), corn, wheat or potato starch, derivatized polysaccharides, so-called gums (for example, xanthan gum), cellulose derivatives.

44 (for example, hydroxypropylmethylcellulose: Klucel ®) or ethylcellulose (Aqualon ®).

In addition, the active ingredient carrier must disintegrate in an optimal and controlled manner in the body in contact with body fluids. Therefore, so-called disintegrants are often added to the decomposition control. Typical compounds for this purpose are corn starches, gelatinized starches and starch modifications. Substances which exert tearing force due to water uptake and swelling that are initiated are also useful.

These include cross-linked polyvinylpyrrolidones (Kollidon CL4), carboxymethylcelluloses and their calcium salts or galactomannans.

Many compounds (e.g. Avicel® and PH102®) have achieved both the necessary mechanical stability and controlled tablet disintegration.

Special starches, including amylosis, have been described as preferred drug carriers for the formulation of tablets (Journal of Pharmaceutical Sciences 55 (1966), 340. However, the amylose Nepol used proves to be disadvantageous (EA Stanley Manufacturing Co., USA) because the drug is released incomplete, and the active ingredient carrier has a high water content (10-12%), so that formulations of hydrolytically labile active ingredients cannot be formulated, in particular cross-linked amylosis (cross-linking degree 15%) is described as a preferred binder (STPPharma Sciences 4 (1994), 329-). 335 and Journal of Controlled Release 15 (1991) 39-46, because of its water-absorbing capacity, it acts as a disintegrant, and in WO 94/21236 cross-linked amylosis is used as a binder and disintegrant (cross-linking degree 25%). however, cross-linking has been shown to be disadvantageous in terms of biocompatibility. Up to 30% by weight of intolerant epichlorohydrin is used, and even a small cross-linking rate of a few percent leads to a rapid decrease in reactivity, so that the remaining residues of the unreacted cross-linking agent must be taken into account.

All commercially available active ingredient carriers containing starches and amylosis utilize sources of plant origin.

The disadvantage here is that these biopolymers, like all naturally occurring substances, exhibit considerable variation in structure and structure and therefore the desired reproducibility and thus uniform product quality is not guaranteed, even with regard to the controlled release of the active ingredient. In the case of natural starches, the content of amylosis and amylopectin varies considerably according to origin. For example, potato starch contains about 20% amyloza and about 80% amylopectin, while corn starch contains about 50% amyloza and about 50% amylopectin. These fluctuations within a single plant community are due to soil standing, fertilizer uptake, seasonal climatic differences, etc. In addition, amylolases may contain 1,4 linked polyglucan with a molecular weight of about 50,000 to 150,000 Dalton and amylopectin, highly branched and 1,4 and 1, 6 is a combined polyglucan having a molecular weight of about 300,000 to 2,000,000 Daltons and a broad molecular weight distribution.

Transitions from highly branched to linear are fluent and fluctuate in the material of plant origin, so sharp separation is practically impossible. In particular, the active compound carriers which still contain amylopectin cause uneven swelling due to the branching, which adversely affects the stability of the carrier. Therefore, amylopectin is usually costly • removed by enzymatic de-branching (Journal of Controlled Release 45 (1997) 25-33 and EP 0499 648 B1 =

No. 5,468,286.

In addition to these stated disadvantages, the wide molecular weight distribution or polymer blends of different spatial configurations, natural polymers contain other components such as low molecular weight compounds, for example fats and oils, which are difficult to separate and which are disadvantageous in further processing and use (e.g. 742). In particular, yield reduction operations must be carried out, in which the impurities cannot be removed in part.

Attempts have also been made to optimize biopolymers, including starches, by altering the starting plants by genetic techniques. WO 94/03049 discloses the production and use of high amylase starches from maize altered by gene techniques. Unevenness and impurities remain a disadvantage.

Reproducibility and quality are strongly dependent on uniformity and purity. In order to guarantee high quality products, these starting substances must be clearly defined and characterized.

SUMMARY OF THE INVENTION It is an object of the present invention to provide a retarding material, avoiding the above disadvantages, which can be used as retarded tablets in controlled release pharmaceutical compositions, preferably for oral administration.

• · • ·

SUMMARY OF THE INVENTION

The problem is solved by using a water-insoluble linear polysaccharide as a retarding material, which is a biocompatible, chemically inert and pressure-stable starting product which, without further additives, allows the controlled release of the active substance. Preferably, a linear water-insoluble poly (1,4-alpha-glucan) is used as such or in the form of spherical microparticles.

Retarded tablets within the meaning of the present invention are in particular tablets, dragees, pills, pellets, compressed shapes, platelets, disc-shaped discs and others whose formulation requires compression. Also included are capsules that are filled with retardant material.

Linear water-insoluble polysaccharides are further considered to be a retarding material.

Linear water-insoluble polysaccharides within the meaning of the present invention are polysaccharides, preferably polyglucans, especially poles (1,4-alpha-D-glucan), which consist of monosaccharides, disaccharides, other oligomers or derivatives thereof.

These are always connected in the same way. Each base unit thus defined has exactly two connections, one to another monomer. The two basic units that form the beginning and end of the polysaccharides are excluded. These base units have only one bond to one other monomer. In three or more linkages (covalent bonding) of one monomer to one other group, preferably to one other carbohydrate unit, this is referred to as branching.

At least three glycosidic linkages then result from each saccharide building unit in the polymer chain.

According to the invention, the branching does not occur at all or only insignificantly so that, in the case of very small branching branches, they are no longer accessible to the usual analytical methods. This is, for example, the case where, based on the total number of existing hydroxyl groups in the number of one hundred hydroxyl groups that are not needed to build a linear polysaccharide, a maximum of five hydroxyl groups are occupied by linkages to other saccharide building units.

The degree of branching is maximum (100% j if each carbohydrate unit has free hydroxyl groups (or other occurring functional groups) of additional glycosidic (or other) bonds to other carbohydrates. The degree of branching is minimal (0%) if on carbohydrates In addition to the hydroxyl groups which condition the linearity of the polymer, no other hydroxyl groups are altered by chemical reaction Examples of preferred water-insoluble linear polysaccharides are linear poly-D-glucans, the type of association being immaterial as long as linearity is maintained within the meaning of the invention. (1,4-alpha-D-glucan) and poly (1,3-beta-D-glucan), the poles (1,4-alpha-D-glucan) being particularly preferred.

If the base unit has three or more connections, it is called branching. This results from the number of hydroxyl groups per 100 basic units which are not involved in the construction of the linear polymer chain and form a branching, the so-called branching degree.

• · ·· ··

According to the invention, the linear water-insoluble polysaccharides have a degree of branching of less than 8%, that is, they have less than 8 branching per 100 basic units.

Preferably, the degree of branching is less than 4% and in particular at most 1.5%.

If the linear water-insoluble polysaccharide is a polyglucan, for example poly (1,4-alpha-D-glucan), the degree of branching at position 6 is less than 4%, preferably at most 2% and in particular at most 0.5% and branching degree in other positions not involved in the linear connection, for example in position 2 or 3 in the case of the preferred poly (1,4-alpha-D-glucan), it is preferably at most 2% and in particular at most 1%.

Particular preference is given to polysaccharides, in particular pol-alpha-D-glucans, which have no branches, or whose degree of branching is so minimal that they are no longer detectable by conventional methods.

According to the invention, the prefix alpha, beta or D applies only to the bonds that form the backbone polymer chain and not to the branching.

Water insolubility within the meaning of the present invention means that there is no recognizable solubility of the compound under normal conditions (room temperature 25 ° C and air pressure 101325 Pa or values deviating by a maximum of 20%).

In the case of the polysaccharides used according to the invention, in particular polyglucans as poles (1,4-alpha-D-glucan), this means that at least 98% of the amount used, preferably the amount

greater than 99.5% is insoluble in water. The concept of insolubility can also be explained by the following observation. When 1 g of the tested linear polysaccharide of 1 ml of deionized water is heated to 130 DEG C. under a pressure of 1 bar, the solution remains stable only for a short time, for a few minutes. After cooling under normal conditions, the substance again falls out. After further cooling and separation on a centrifuge, taking into account the test losses, at least 66% of the amount used can be recovered in this way.

Linear, water-insoluble polysaccharides are preferably used in the present invention, which can be obtained by means of biotechnological or gene manipulation methods of general definition. One particularly preferred embodiment of the invention described herein is production by a bitechnological process, in particular by a biocatalytic process.

Linear polysaccharides produced by biocatalyst (also biotransformation) within the scope of the present invention means that a linear polysaccharide is produced by catalytic reaction of monomeric building blocks as oligomeric carbohydrates, for example mono- and / or disaccharides, using a so-called biocatalyst, usually an enzyme, under appropriate conditions . In particular, polyl- (1,4-alpha-D-glucan) is usually produced by polysaccharide synthases and / or starch synthases and / or glycosyltransferases and / or alpha-1,4-glucantransferases and / or glycogen synthases and / or amylocucrase and / or phosphorylase.

Linear polysaccharides from fermentation are also contemplated. These are linear polysaccharides which can be obtained by fermentation processes using naturally occurring organisms, such as fungi,

9 4 4

4 • • 4 4 • • 4 9 • • 4 4 4 9

Algae or micro-organisms or using non-natural organisms obtained by modification of natural organisms, such as fungi, algae or micro-organisms, by gene manipulation of a general definition.

Further, the linear polysaccharides for the production of the retarded tablets described in the present invention can be obtained from non-linear polysaccharides which contain branching by treating with the enzyme and after cleavage (for example with enzymes such as amylase, isoamylase, gluconohydrolase, pullulanase and others). branching to obtain linear polymers.

The molecular weights M _{w of the} linear polysaccharides used according to the invention can vary within a wide range from 10 g / mol to 10 ⁷ g / mol, preferably the molecular weights M _w range from 2 x 10 g / mol to 5 x 10 g / mol, in particular 3 x 10 g / mol to 2 x 10 g / mol. For the preferred linear polysaccharide poly (1,4-alpha-D-glucan) used, the corresponding range is used.

The molecular weight distribution or polydispersity M _w / M _n can vary within wide limits according to the process for producing the polysaccharide. Preferably, a polydispersity of 1.01 to 50, particularly preferably 1.5 to 15 is used. Here, the polydispersity increases with the bimodal molecular weight distribution, which does not adversely affect the formulation of the tablets.

The mixtures of the linear polysaccharides according to the invention, even in the form of microparticles with non-linear polysaccharides, are not excluded. Controlled release of an active substance is understood to mean that the active substance, taking into account the statistical variation corresponding to the circumstances after a certain time and / or interval t, is defined as:

9 9 ··· ····

99 9 9 9 9 999 9 9 9

9 · · · 9999

999 9 «··· ·· ·· ·· releases in a dose advantageous to the biological organism.

This definition also includes extremes. On the one hand, the spontaneous release of all active ingredients in the formulation over a period of time reduced to zero, on the other hand the minimum amount / dose required to achieve a therapeutic effect for a long, even infinite time, at least a certain time required to release all active ingredients present in the formulation.

Therefore, the retarded formulation herein is referred to as a synonym for a depot formulation or a delayed release formulation. Any biologically active substance and combination of substances in the broadest sense of the term (with an emphasis on the human and veterinary field), in particular for medical indication, shall be considered as active substance. These are in particular: analgesics, anginal preparations, antiallergics, antihistamines, antiinflammatoria, bronchodilators, bronchospasmolytics, diuretics, anticholinergetics, anti-adhesion molecules, cytokine modulators, biologically active endonucleases, recombinant human DNases, antagonists, neurotransmitterin intestines, neurotransmitterin, neurotransmitterin analeptics, analgesics, local anesthetics, narcotics, antiepileptics, anticonvulsants, antiparkinsonian drugs, antiemetics, hormone regulating or stimulating compounds, circulatory regulating or stimulating compounds, respiratory regulating or stimulating compounds, vitamins, trace elements, antioxidants, cytostatics, antostatics , antiinfectives, immunomodulators, immunosuppressants, antibiotics, proteins, peptides, hormones, growth hormones, growth factors, xanthines, vaccines, steroids, beta2-mimetics.

• · w «· r

• 9 · 9 · 9 · · «·» · »• * 9 9 9 9 9 9 9 9 9

I · I «t ·· ··

A therapeutic effect within the meaning of the invention means that a therapeutically effective amount of the active ingredient reaches the desired target site, exerts its effect there and causes a physiological reaction. Palliative and / or curative effect is included.

Biocompatible in the sense of the invention means that the polysaccharides used undergo complete biodegradation and are not enriched in the body. Biodegradation is always understood to mean an in vivo process which leads to degradation or degradation of the polymer. Also hydrolytic or enzymatic processes are included in this field. Last but not least, the natural character of the polysaccharides used is also important for the biocompatibility of polysaccharides and their degradation products (metabolites). Therefore, the polysaccharides used according to the invention are particularly suitable for therapeutic, diagnostic or prophylactic use. The term pharmaceutically acceptable within the meaning of the present invention means that the active ingredient carrier, excipient or so-called excipient can be accepted by the living being without producing significant side effects in the body.

The preparation of the tablets is carried out by mixing the starting components, the linear polysaccharide being homogenized with the active substance by known methods and homogenized, for example by means of ball mills. The active ingredient may have a concentration of up to 50%, preferably between 1 and 20%, particularly preferably between 5 and 15%. Other conventional excipients and additives may be used. The sum of the active ingredient and the polysaccharides according to the invention in the total composition (including optional excipients and additives) should be at least 50%, preferably 70 to 100%, particularly preferably 85 to 98%. The composition of the excipients may vary within wide limits, the ratio of which depends on the interactions with the active ingredient and the water-insoluble linear polysaccharide.

Solvents can be used as an auxiliary in the production of tablets as well as in the previous mixing, with readily volatile solvents being preferred.

The basic body of the polysaccharides according to the invention for the production of tablets can be an amorphous or crystalline form or grains as they arise directly in the synthesis, or also microparticles as described in the patent application (DPA,

Az: 197 37 481.6 A simple mixing process is preferably used to produce the raw material or the raw mixture. The method of making tablets may affect the properties of the tablet. For example, it is possible to combine the active ingredient with the polysaccharide body by spraying, for example in a fluidized bed or by coating the suspension on the polysaccharides used according to the invention. Suction processes in which the porous form of the microparticles is used to absorb the active substance in solution (sponge character) or spray-drying techniques are useful in this case. Here, the solution, suspension or emulsion of the linear polysaccharide and the active ingredient is dried by known spray-drying techniques. In the case of solutions, the corresponding organic solvents are used. Higher temperatures or pressures, as well as supercritical methods, can help in this in order to achieve the required solubility in a short time.

The pressures used during tablet manufacture may vary within wide limits. The pressure variations can be specifically used in the case of the polysaccharides described according to the invention to achieve an additionally positive retarding effect. The pressures can vary within wide limits from 1 MPa to 10 ³ MPa (10 ^ Pa · 4 · 4 · 4 · 4 · 4 ·

4 44 * · · · · · 4 · 4 ·

4 · · · ·

444 44444 4 * «· = 1 bar). Preference is given to pressures in the range from 10 MPa to 300 MPa, particularly preferably from 100 MPa to 250 MPa.

The following examples and figures are intended to illustrate the invention in more detail without limiting the scope of the products and methods of the invention.

DETAILED DESCRIPTION OF THE INVENTION

The following examples relate in particular to the production of microparticles as described in the patent application (DPA, AZ: 197 37 481.6), which is hereby incorporated by reference. Furthermore, a particularly advantageous method for producing polyl- (1,4-alpha-D-glucan) in WO 95/31553 is described.

Example 1

Production of in-vitro polyl- (1,4-alpha-D-glucan) by biocatalytic process using amylocucrase enzyme

A 10 L 20% sucrose solution is placed in a sterilized 15 L vessel (steam sterilization). The amylocucrase-containing enzyme extract obtained by fermentation is added all at once to the sucrose solution. The enzyme activity is 16 units (1 unit corresponds to a reaction of 1 pmol sucrose per minute per mg of enzymes). The apparatus is also provided with a sterilized stirrer KPG. The vessel was sealed and kept at 40 ° C and stirred. After some time a white precipitate formed. The reaction was complete after 180 hours. The precipitate is filtered off and washed repeatedly to remove the low molecular weight sugars. The residue remaining on the filter is dried at a temperature of between 30 and 40 ° C in an oven under vacuum, using a vacuum. • Diaphragm pumps (Vacuubrand GmbH & Co., CVC 2). The weight is 685 g (69% yield).

Example 2

Characterization of the polyl- (1,4-alpha-D-glucan) synthesized by the amylocucrase of Example 1 by gel permeation chromatography mg of the polyl- (1,4-alpha-D-glucan) of Example 1 was dissolved in dimethylsulfoxide (DMSO, (Riedel-de-Haen) and filtered (2 mm filter). A portion of the solution was injected onto a gel permeation chromatography column. DMSO was used as the eluent. Signal intensity is measured using an RI detector and evaluated against a pullulan standard (Polymer Standard Systems). The flow rate was 1.0 minute.

From the measurement of apparent number average molecular weight (M _{n) of} 2700 g / mol and a weight-average molecular weight (M _w) of 11,700 g / mol. This corresponds to a dispersion of 4.3.

Example 3

Production of microparticles of poly- (1,4-alpha-D-glucan)

400 g of poly (1,4-alpha-D-glucan) was dissolved in 2 l of dimethylsulfoxide (DMSO, p.a. from Riedel-de-Haen) at 60 ° C over 1 hour. It is then stirred for one hour at room temperature. The solution was added dropwise to the 20 L of double-distilled water with stirring via a dropping funnel over 2 hours. The batch is stored at 40 ° C for 40 hours. A fine suspension is formed. The particles are separated by first decanting the residue. The pellet is removed and centrifuged in small portions (RC5C ultracentrifuge: 5 minutes at 5000 rpm). The solid residue is washed a total of three times with distilled water twice and centrifuged again. The solid was combined and the suspension of about 1000 ml was freeze-dried (Christ Delta 1-24 KD). 283 g of a white solid are isolated (Example 3a: 71% yield). The combined residues were stored at 18 ° C overnight. The processing is carried out as described. An additional 55 g of a white solid was isolated (Example 3b: 14% yield). The overall yield is 85%.

Example 4

Desulphurization of the microparticles of Example 3

Proceed as described below to separate the dimethylsulfoxide remaining in the particles. 100 g of the amylase particles of Example 9 are introduced into 1000 ml of deionized water. The batch is left for 24 hours with gentle movement. Particle separation was performed as described in Example 9 (RC5C ultracentrifuge: 15 min each, 3000 rpm). After freeze-drying, 98.3 g (98% yield) are obtained. Determination of sulfur by elemental analysis gives the following values (combustion test method and IR detection):

Sulfur content of the particles of Example 2: 6% ± 0.1%

Sulfur content of the particles of Example 3: <0.01%

Example 5

Examination of the microparticles of Example 3 by electron

• · · · · · · · · · · · · · · · · ·

Microscopy

Electron Scanning Microscope (REM) images (Camsan S-4) are used to characterize the particles. Figures 1 and 2 show particle images showing that they are spherical, very uniform particles in terms of shape, size and surface roughness.

Example 6

Examination of the particle size distribution of Example 3

To characterize the particle size distribution of Examples 1 and 9, a Mastersizer (Malvern Instruments) was tested. Testing is performed in Fraunhofer mode (evaluation: multimodal, count) with a density of

1.080 g / cm ^{&lt; 3} &gt; and a volume concentration in the range of 0.012% to 0.014%.

Table 1

Characterization of the microparticle diameter of Example 3

TA O V Diameter Particle distribution Ex. Dn * · (mm) Dw ² (mm) dw / dn ^ d (10%) ⁵ (mm) d (50%) ⁵ mm d (90%) ⁷ (mm) 3a 1,664 4,184 2,541 0,873 1,504 2,624 3b 0,945 2,345 2,481 0.587 0.871 1,399

¹ d d diameter average weighted d / d diameter average ⁴ d (10%) ⁵ d (50%) ⁶ d (90%) particle diameter dispersion% of all particles has smaller diameter than stated% of all particles has smaller diameter than stated the% value of all particles has a smaller diameter than said value

Example 7

General Method for Making Tablets from Polypeptides (1,4-alpha-D-Glucan)

In a ball mill (Retsch MM2000), 270 mg of the tablet excipient (polyl- (1,4-alpha-D-glucan)) and 30 mg of active ingredient are milled for 10 minutes at an amplitude of 100 (manufacturer's data). 250 mg is withdrawn from the homogenised batch and transferred to a pressing tool (Perkin Elmer; punch diameter 13 mm). The press tool is placed under the press (Perkin Elmer, hydraulic press). Subsequently, the mass is pressed at a pressure of 2 t for 10 minutes. After depressurizing the apparatus, the finished tablets are carefully removed and stored for further characterization, for example stability measurement or release testing.

Further, tablets of known tablet-making materials (comparative examples) such as microcrystalline cellulose (Avicel), potato starch (Toffena®-Sedtárke) and polyacrylates (Eudragite® Rhoma) will be prepared for comparison purposes.

· · 4 4 4 4 4 4 4 4 4 *

4 4 4 4 4

4449 44 444

4 4 4 4

44 * 4 4 4 4 * 44 4 4 4

4 ♦

4 4

4 4

4

Example 8

Determination of drug release over time

The disintegration of the tablets produced according to Example 7 is determined as follows. One tablet is placed in 25 ml of water (deionized water) in a 50 ml Erlenmeyer flask. The throat is covered with a parafilm. The flask is mounted on a shaker (IKA Labortechnik; KS 125 basic). The shaker is operated at a setting of about 150 / minute. After a period of time, about 1.5 ml of the remaining solution is sampled.

From this volume, enough is transferred to a disposable cuvette (Sarstedt No. 67.741) and measured on a spectrometer (Kontrom Instruments, Uvikon 860). The absorption maxima indicated applies to individual active substances or model substances.

Example 9

Absorption maxima of other active substances

The absorption maxima of the other active substances are determined as in Example 10. All the active substances mentioned give comparable results in observing the retarding effect, from which conclusions can be drawn from the viewpoint of the versatility of possible use.

»* * * * * # # # # # # # # # # # # # # # # # # # # # # # # # # # # ··· 9 · 9 · 9 · 9 · 9 ·

Table 2

Absorption maxima of the different active substances examined

Active substance Absorbent maximum Vitamin B 12 549 nm Theophyllin 271 nm ³ TM Ramorelix 276 nm Caffeine 272 nm ³ Iloperidon 274 nm ³ Buserelin ^ 278 nm ³ Minoclinhydrochlorid 278 nm ³ Tetracycline hydrate 269 nm ³ Phenylephrin 272 nm ³

Structures a: 5-oxo-L-prolyl L-histidyl-L-tryptophy 1-lasers L-torosyl-β-tert-butyl-D-seryl-L-leucil L-arginyl-N-ethyl-L-prolinamide ^ Cuvette single use (Sarstedt No. 67.741) on ^ Quartz cuvette (Hellma, Suprasil)

Example 10

Capture of the calibration curve for the active substance on the example of model compound vitamin B 12% stock solution is prepared by weighing 100 mg of vitamin B 12 into a 10 ml volumetric flask and making up to the brand mark with deionized water. A dilution series of 0.005%, 0.01% and 0.02% is prepared from it and measured in spectra20.

4 ► 4> 4 • 44 4

4 4

4 4 • • 4 4 4 of the meter (Kontrom Instruments, Uvikon 860). Extinctions are read at an absorption maximum lambda = 549 nm. Additional measuring points are required if deviations from the line are apparent. These calibration lines serve as a starting point for determining the concentration in the remainder of the drug release experiment. The calibration curves of other active substances and model substances are read in an analogous manner. A graphic representation of the calibration for vitamin B 12 (concentration-dependent extinction in water) is shown in Figure 3.

Example 11

Attempts to release vitamin B 12 from tablets made with polyl- (1,4-alpha-D-glucan) and microparticles therefrom

As described in Example 10, the extinction of the residue from the in vitro release experiment was measured after a period of time. To obtain appropriate concentrations from the extinction measured by the calibration curve, it may be necessary to dilute the residues at a ratio of 1:10. This factor is then taken into account accordingly.

The values are summarized in Table 3. The corresponding figure 4 shows the values graphically. The maximum possible concentration - 0.1% according to Example 8, is compared to 100% for a clearer illustration, so that an assessment of the degree of release after which time is achieved is possible.

··· * · «· · · · · · · · · ·

4 4 4 • 4 4

Table 3

Concentration values of the aqueous residue versus time for the tableting auxiliaries described in accordance with the invention are the poly (1,4-alpha-D-glucan) and the microparticles made therefrom.

The release of vitamin B 12 from tablets made with various excipients is shown:

a) poles- (1,4-alpha-D-glucan)

(b) microparticles of poly (1,4-alpha-D-glucan)

Auxiliary for the manufacture of tablets Clock ^ A ² A ² A ² B ³ B ³ B ³ 0 0 0 0 0 0 0 0.5 5.44 2.33 4.91 3.97 3.52 3.14 1 6.74 4.87 5.94 4.03 3.74 3.89 2 11.45 7.57 8.18 6.52 5.72 5.63 3.5 24.45 15.32 19.57 14.22 9.43 11.19 6 35.26 26.11 30.36 20.37 17.23 18.92 8 41.14 27.20 36.38 23.71 17.77 22.94 10 43.28 34.78 36.38 26.12 21.90 22.94 24 60.90 55.02 67.08 33.23 31.07 39.92 32 63.83 61.21 74.96 35.70 28.13 46.83 48 75.27 64.30 87.17 38.18 33.38 53.17

^ Release time in hours ² Poly- (1,4-alpha-D-glucan) ^ Polio- (1,4-alpha-D-glucan) microparticles ·· 9 99 99 · 9 • 9 · 9 9 9 ···

999 9 »9 999> 9 9 * 999 999 · 99 99 99999 99 99 9

Example 12

Attempts to release vitamin B 12 from tablets made from micTM rocrystalline cellulose (Avicel) and potato starch Toffena ™) (comparative examples)

The results shown in Table 4 were measured and calculated as described in Example 11. Figure 8 shows the results compared to the tableting excipient described according to the invention poly (1,4-alpha-D-glucan) against the comparative microcrystalline cellulose substance. (Avicel)

TM and potato starch Toffen ¹ ). The retardation effect is clearly recognizable here.

• 9 ·

9, 9, 9, 9, 9, 9, 9, 9

9

9 9 9 • 9 99 99

9 9 9 9 9 · 9 9 9 · * 9 9 9 · 9 9

9 9> 9 ·

9 ·· 9 *

Table 4

Aqueous Time Concentration Values for TM Comparative Substances Microcrystalline Cellulose (Avicel) TM and Potato Starch (Toffena)

Auxiliary for the manufacture of tablets Clock ^ Avicel ² Avicel ² Avicel ² Starch ³ Starch ³ Starch ³ 0 0 0 0 0 0 0 0.5 6.99 6.51 4.81 7.99 12.18 93.97 1 13.38 14.84 17.68 74.65 33.54 74.34 2 34.31 22.30 26.12 77.28 39.57 77.28 3.5 52.40 31.53 37.51 76.51 65.07 91.14 6 65.53 43.89 50.54 78.98 69.86 84.70 8 75.12 52.40 57.50 78.98 77.43 80.53 10 77.13 67.23 57.50 78.98 84.08 80.53 24 78.36 84.70 95.05 83.15 87.48 87.79 32 72.80 81.92 100 ALIGN! 80.53 85.32 87.48 48 79.75 84.39 100 ALIGN! 84.08 82.07 87.48

Release time in hours

TM Avicel microcrystalline cellulose TM Toffena TM potato starch

Figure 5 shows the release of vitamin B 12 from tablets with various excipients:

a) poles- (1,4-alpha-D-glucan)

(b) microparticles of poly (1,4-alpha-D-glucan)

TM

(c) Avicel ^{1 1} microcrystalline cellulose (comparative example);

99 99

9 9 9 ·

9 9 9 9 9

9 9 · · ·

(d) Toffena TM potato starch (comparative example) (For clarity, the mean values given in Tables 3 and 4 were calculated).

Example 13

Attempts to release theophylline from tablets made from polyl- (1,4-alpha-D-glucan) and microparticles therefrom

Attempts to release theophylline from tablets consisting of various excipients for the manufacture of tablets are carried out as described in Example 12. Polymers (1,4-alpha-D-glucan), which is obtained directly from the biocatalyse process corresponding to processing (cf. Example 1), as well as microparticles of poly (1,4-alpha-D-glucan). The results are shown in Figure 6. A duplicate assay is always performed to demonstrate high reproducibility of the results.

Example 14

Attempts to release theophylline from tablets made from TM

(c) microcrystalline cellulose (Avicel),

(d) Eudragit RS TM; _and

TM

e) Eudragit RL ¹ (comparative examples)

Attempts to release theophylline from tablets consisting of comparative excipients for the manufacture of tablets were carried out as described in Example 14. Microcrystalline cellulose (Avicel TM), Eudragit RS TM _and Eudragit RL.TM were used as tablet excipients. in Figure 6. A duplicate assay was performed.

• 9 9 9 9 9 9 999 99 9 9 9 9

Figure 6 shows the release profiles of Example 13 in comparison.

Figure 6 shows theophylline release from tablets with different excipients:

(a) poles (1,4-alpha-D-glucan),

(b) microparticles of poly (1,4-alpha-D-glucan),

(c) microcrystalline cellulose (Avicel ^T ^),

(d) Eudragit RS a

e) Eudragit RL.

Example 15

Release attempts from theophylline tablets consisting of poly- (1,4-alpha-D-glucan) as well as poly (1,4-alpha-D-glucan) microparticles and microcrystalline cellulose (Avicel ®) (comparative example) in artificial gastric šfávě

Attempt to release theophylline from tablets with the excipient for the production of poly (1,4-alpha-D-glucan) tablets (including poly (1,4-alpha-D-glucan) microparticles and microcrystalline cellulose TM) (Avicel ¹¹¹ ) in artificial gastric juice is performed as described in Example 8 (artificial gastric juice: 2 g sodium chloride, 3.2 g pepsin, 7 ml concentrated hydrochloric acid (HCl), made up to 1 liter of total volume with deionized water). using a medium that corresponds to natural conditions, a reproducible mode of release retarding effect can be observed.

Figure 7 shows theophylline release from tablets of various excipients:

(a) polyl- (1,4-alpha-D-glucan),

··

(b) Poly (1,4-alpha-D-glucan) microparticles; and

ΤΓΜ

c) microcrystalline cellulose (Avicel) (comparative example).

Example 16

Release attempts from theophylline tablets consisting of polyl- (1,4-alpha-D-glucan) and microparticles made therefrom

TM

The Ramorelix release experiments were carried out in the same manner as in the examples so far. Ramorelix ™ is an LHRH antagonist with the following amino acid sequence (structure):

1-N- (acetyl-3- (2-naphthyl) -D-alanyl-p-chloro-D-phenylalanyl-Dtrypthophyll-L-seryl-L-tyrosyl-O- (6-deoxy-alpha-L-mannopyranosyl)) -D-seryl-L-leucyl-L-arginyl-L-prolyl) semicarbazide acetate. However, artificial gastric juice was used as the release medium instead of deinized water (E-water). The formula of artificial gastric juice is: 2 g of sodium chloride, 3.2 g of pepsin, 7 ml of concentrated hydrochloric acid (HCl _and q), made up to one liter of total volume with deionized water. The pH of the solution is 1.2.

Example 17

TM

Release experiment from Ramorelix tablets consisting of microcrystalline cellulose TM (Avicel) (comparative example)

The release experiments were carried out as described in Example 8. Artificial gastric juice is used as the release medium.

Figure 8 shows the release of Ramorelix ™ from tablets with different excipients:

··· · ·

• ··

(a) poles (1,4-alpha-D-glucan),

(b) Poly (1,4-alpha-D-glucan) microparticles; and

c) microcrystalline cellulose (Avicel ™) (comparative example).

Example 18

Determination of solubility of polysaccharides

100 mg of poly- (1,4-alpha-D-glucan) are taken up in 5 ml of double-distilled water. The reaction vessel is slowly warmed with stirring (magnetic stirrer). It gradually warms up and visually observes in a 20-step program. No changes are observed at temperatures of 40 ° C, 60 ° C, 80 ° C and 100 ° C. According to these observations, the compound may be classified as water insoluble.

Example 19

Determination of solubility of polysaccharides and classification according to German Pharmacopoeia (DAB)

564 mg of poly (1,4-alpha-D-glucan) is heated in about 0.5 L of double-distilled water at a pressure of 100 psi and 130 ° C for 1.5 hours in an autoclave (Certoclav apparatus). The weight of the reaction vessel was determined beforehand. The apparatus is then depressurized and cooled at room temperature. Weigh the content. Corresponds to 501.74 g. After a further 24 hours, it is centrifuged and decanted. The solid residue was dried and weighed: 468 mg. The dissolved fraction of 96 mg is calculated from this. Based on the solvent used, it is calculated that 5 226 mg of water is required per mg of poly (1,4-alpha-D-glucan). According to the classification of the German Pharmacopoeia, this implies classification as • 99 · 9 9

• 9 9 9 9 9

Very poorly soluble because between 1,000 and 10,000 parts of solvent is needed to dissolve 1 part of the substance.

This is from 7 classes to classify solubility (from very easily soluble (class 1) to practically insoluble (class 7) class number 6).

Example 20

Determination of solubility of polysaccharides and classification according to German Pharmacopoeia (DAB)

The experiment is carried out as described in Example 19. The only difference is the cooling process which is applied after autoclaving and cooling to room temperature. The mixture is stored at 5 ° C for 3 hours.

Weigh 526 mg of poly (1,4-alpha-D-glucan) to about 480 ml of double-distilled water. After heat treatment, the balance was 468.09 g. The dried sediment weighed 488 mg. Accordingly, 38 mg of poly (1,4-alpha-D-glucan) was transferred to the solution.

This corresponds to a ratio of 1 mg of substance to 12,318 parts of solvent. Accordingly, the substance is classified in Class 7 of the German Pharmacopoeia in this treatment and is classified as practically insoluble because more than 10,000 parts of solvent are needed per part of the substance.

Claims (12)

PATENT CLAIMS Retarded tablets comprising wholly or partially at least one water insoluble linear polysaccharide as a retardant material. Retarded tablets according to claim 1, comprising wholly or partially at least one water-insoluble linear polysaccharide which is produced by a biotechnological process. Retarded tablets according to claim 1, comprising wholly or partially at least one water-insoluble linear polysaccharide which is produced by a biocatalytic process. Retarded tablets according to claim 1, comprising wholly or partially at least one water-insoluble linear polysaccharide which is produced by a fermentation process. 5. Retarded tablets obtainable from linear water-insoluble polysaccharides. Retarded tablets according to any one of claims 1 to 5, comprising wholly or partially at least one water-insoluble linear (1,4-alpha-D-glucan). Retarded tablets according to one or more of Claims 1 to 6, characterized in that the water-insoluble linear polysaccharides are wholly or partially in the form of microparticles. 8. Retarded tablets according to one or more of Claims 30 4 * 4 4 4 · 4 · 4 · 4 · 4 · 4 · 4 · 4 · 4 · 4 · 44 · 4 · 4 4444 44 44444 44 44 years 1 to 7, characterized in that they are biocompatible and / or pharmaceutically acceptable. Use of retarded tablets according to one or more of claims 1 to 8 for the controlled release of active substances. Retarded tablets according to one or more of claims 1 to 9, containing one or more active substances and optionally further additives and excipients. Retarded tablets according to one or more of Claims 1 to 10, characterized in that they exert a therapeutic effect as pharmaceutical preparations. Use of retarded tablets according to any one of claims 1 to 11 as formulations, preferably as oral dosage forms.