DE2423811A1 - PROCESS FOR THE PRODUCTION OF FREEZE-DRIED 2,2'-ANHYDRO-1-BETA-D-ARABINOFURANOSYLCYTOSINE PREPARATIONS AND INJECTION PREPARATIONS - Google Patents

Description

⁰ Process for the production of freeze-dried 2,2'-anhydro-1-ß-D-arabinofuranosylcytosine preparations and injection preparation "

Priority: May 17, 1973, Japan, No. 54 145/73

The invention relates to a process for the production of freeze-dried 2.2 ^l -anhydro-1-ß-D-arabinofuranosylcytosine preparations, hereinafter also referred to as cyclocytidine preparations for short, which are characterized by improved solubility. The invention also relates to a. Injection preparation which consists of an aqueous solution of the freeze-dried preparation produced according to the invention.

• As is known, cyclocytidine shows an activity against leukemia in mice (cf. e.g. A, Hoshi et al., Gann (Cancer), Vol. 62 (1971), p. 145) μΜ it is characterized by a low toxicity » compared to 1-ß-D-arabinofuranosylcytosine, which is used for was used for the same purpose; see e.g. H. Hirayama et al., Pharraacometrics, Vol. 6 (1972), p. 1259. It is also

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knows that cyclocytidine has an excellent cytostatic effect against the L1210 tumor and other experimental tumors having; See, e.g., Y. Sakai et al., Jan. J. Clin. Oncol., Vol. 6, Issue 2 (1972), pages 57 to 64.

However, cyclocytidine has the disadvantage that it is not sufficiently stable in aqueous solution. In an aqueous alkaline solution, the 2,2'-bond of the cyclocytidine is easily cleaved with the formation of 1-ß-D-arabinofuranosylcytosine. For the production of medicaments which contain cyclocytidine, which is unstable in aqueous solution, a powder is therefore usually used which contains the compound i n /
the / contains freeze-dried form. For the production of sterile preparations, such as injection preparations, the technical manufacture of the preparations in the form of freeze-dried products would therefore be advantageous.

When carrying out the freeze-drying of cyclocytidine in the usual way for the production of drugs, the production of injectables, and when examining the properties of the product, it was found that that when the freeze-dried product is dissolved in water, particles are undissolved at a slow rate of dissolution remain and a cloudy solution is obtained.

Studies have shown that when a dry preparation with a water content of at most about 2 to 3 percent by weight in water, e.g. up to a concentration of up to 10 percent by weight, a cloudy solution is obtained. It was also found that when dissolving a dry preparation_j

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rait a water content of at least about 3 percent by weight in water, the solution obtained is clear. However, such dry preparations with a high water content have insufficient stability. It was also found that when freeze-drying at a relatively low temperature by reducing the heat input, for example when drying at a plate temperature of the freeze-drying chamber of about 0 ^° C., the solution obtained from the product is not cloudy, even if the water content of the end product is at most about 1 Weight percent is. When freeze-drying at such low temperatures, however, very long drying times are required, which makes the process uneconomical.

The invention is therefore based on the object of a powerful Process for the production of freeze-dried cyclocytidine preparations to create, which are characterized by an improved solubility, and which are suitable for parenteral administration, particularly suitable for subcutaneous, intramuscular and intravenous injection. This object is achieved by the invention solved.

The invention thus relates to a process for the production of freeze-dried 2,2 »-anhydro-1-ß-D ~ arabinofuranosylcytosine preparations, which is characterized in that a 2.2 ^l -anhydro-1-ß-D- arabinofuranosylcytosine and

(1) a saccharide, (2) polyvinylpyrrolidone, "(3) sodium carboxymethyl cellulose or (4) containing a nonionic wetting agent or a mixture of at least two of these compounds l_aqueous solution freeze-dried, which the components (1),

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respectively
(2) and (3) in an amount νοη'θ, 1 to 5 percent by weight and component (4) in an amount of 0.01 to 0.05 percent by weight, based on the cytosine compound.

The term "improved solubility" means that upon dissolution practically everything dissolves from 1 g of freeze-dried cyclocytidine preparation according to the invention in 10 ml of water. Of the The term "aqueous solution" means water and other media suitable for parenteral administration, for example a physiological saline solution, an aqueous glucose solution and Ringer's solution. The invention is described below with reference to Use of water * as a solvent is explained.

In practice, the method according to the invention is carried out in such a way that that the aqueous solution which contains the aforementioned ingredients in the specified amounts in a container for Injection preparations, for example an ampoule or another glass vessel, are filled and the solution is freeze-dried.

Examples of the saccharides which can be used in the process according to the invention are monosaccharides, disaccharides and polysaccharides, Pentoses and hexoses, as well as the corresponding sugar alcohols. Specific examples of preferably used saccharides are Monosaccharides such as fructose, sorbitol, xylitol, mannitol and glucose, disaccharides such as lactose, and dextrans with a middle Molecular weight of about 20,000 to 250,000. Sorbitol is particularly preferred.

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Specific examples of those that can be used in the process of the present invention are non-ionic surfactants

(a) Sorbitan fatty acid ester-ethylene oxide adducts, such as sorbitari- ' monooleate-polyoxyethylene (20) adducts, for example polysorbates 80 and Tween 80, and sorbitan monolaurate polyoxy

. ethylene (20) derivatives, such as NIKKOL TL-10,

(b) Ethylene oxide adducts with hydrogenated castor oil (castor oil), such as NIKKOL HCO-40, 50 and 60, with 40 ,. 50..or. 60 ethylene oxide units in the molecule and

(c) Ethylene oxide-propylene oxide block copolymers, such as Pluronic L44, L62, L64 and L68.

As indicated above, the saccharide, the polyvinylpyrrolidone, the sodium carboxymethyl cellulose and the nonionic wetting agent can be used alone or as a mixture of at least two of these compounds. A specific example of a polyvinylpyrrolidone which can be used according to the invention is Piasdon, and a specific example of a carboxymethyl cellulose which can be used corresponds to the quality of US Pharmacopoeia XVIII. Typical examples of mixtures of the compounds are combinations of polyvinylpyrrolidone and glucose and sorbitol and a nonionic wetting agent. The proportion of these additives is about 0.1 to 5 percent by weight, based on the weight of the cyclocytidine, regardless of whether these compounds are used alone or in a mixture. The preferred amount of the particular compound or compounds used depends to some extent on the type of additive used. Beispielsweise'ist the preferred amount is 2 to 5 weight percent in the FAI ^- Ie of saccharides, 0.1 to 5 weight percent,

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preferably 0.3 to 5 percent by weight, in the case of polyvinylpyrrolidone, 0.1 to 2 percent by weight in the case of sodium carboxymethyl cellulose and 0.01 to 0.05 percent by weight in the case of the non-ionic wetting agent. If a combination of, for example, polyvinylpyrrolidone and glucose is used the polyvinylpyrrolidone preferably in an amount of 0.3 to 4 percent by weight and the glucose preferably in an amount from 1 to 4 weight percent used.

According to the method according to the invention, a mixture of cyclocytidine and at least one of the aforementioned compounds is first dissolved in water. Water is generally used in about 8 to 20 times the amount by weight of the cyclocytidine. Preferably about 10 ml of water per gram of cyclocytidine are used. The use of more than about 20 times the amount by weight of water is uneconomical for the subsequent freeze-drying stage. If smaller amounts of water than about 8 times the amount by weight are used, it is more difficult to dissolve the mixture. The constituents are preferably dissolved in water at room temperature, ie at about 20 to 30 ^° C. Cooling is not required. Since cyclocytidine tends at temperatures above about 60 ° C to decompose the constituents to be dissolved at temperatures below about 60 ⁰ C. Temperatures below the freezing point of water are not suitable. The cyclocytidine and the additive or additives can be dissolved in water in any order, ie the cyclocytidine and the additive or additives can be dissolved in water separately or at the same time.

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mm ( «a»

Injection grade distilled water is generally used to prepare the solution. The solution obtained is usually filtered through a bacterial filter with a pore size of about 0.22 microns or a Seitz filter and inserted into a conventional sterile ampoule or other glass container. Each ampoule or each glass vessel is preferred

din
filled with a single dose of Cycüocyfci /. The cyclocytide volume per glass vessel is about 100 to 500 mg. The solution is then freeze-dried in the glass vessel or the ampoule in a conventional manner. For this purpose, the ampoules or glass vessels containing the solution of the constituents are placed directly on a plate or in trays on a plate in a freeze-drying chamber. After the chamber has been closed, the ampoules or glass vessels are cooled at normal pressure. The cooling temperature depends to a certain extent on the room temperature and the temperature of the cooling unit. Usually the system is cooled so much that the temperature is about -40 to -45 ° C. As soon as the temperature of the contents of the ampoules or glass jars has reached this range and the electrical conductivity has reached fourth 0, which indicates that the solution is completely frozen, the freeze-drying chamber is gradually evacuated and drying commenced. The evacuation can be carried out to a pressure of about 0.01 torr or less, which is generally established within a few minutes. However, this depends on the size of the freeze-drying chamber and the pump capacity. As a result of the heat of sublimation, the temperature of the contents of the glass vessels or ampoules is reduced. The system is therefore heated so that the temperature of the plate j

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is maintained at a fourth of about 20 to 3O ⁰ C. In this

In this case, the system is preferably kept at a value of about 30 ° C. in the initial stage of drying. Once the drying curve if the temperature rises, the temperature is reduced. The breadth of temperature control is determined by the drying curve of the recorder controlled. If the temperature of the contents of the ampoules or glass jars coincides with the temperature of the plate, the drying is finished. After complete drying, the pressure in the freeze-drying chamber is increased by introducing it brought to normal pressure by sterile air and the ampoules or glass vessels are removed from the freeze-drying chamber removed, melted or closed with sterile rubber stoppers. The water content of the product in the ampoules or glass vessels is at most about 3 percent by weight, generally about 0.8 to 3 percent by weight.

The process according to the invention is carried out in all stages under sterile conditions. Of course you can the cyclocytidine and the additives used do not contain any pyrogens. The medicinal preparations produced according to the invention usually become injectables just prior to administration with ampouled sterile distilled water processed and injected immediately.

The particle size of the cyclocytidine preparations produced according to the invention is of similar size to the particle size produced by conventional freeze drying processes Preparations. When dissolving the preparations produced according to the invention in an aqueous medium in high concentration for in-,

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For injection purposes, the preparation dissolves very quickly with formation a clear solution with excellent stability.

For example, the rate of dissolution of a conventional lyophilized cyclocytidine preparation is 45.8 seconds and the resulting solution has a permeability of 80.3 percent. The speed of resolution of the Cyclocytidine preparation produced according to the invention, the 1 percent by weight cyclocytidine and 2 percent by weight sorbitol, based on the weight of the cyclocytidine, is 7.4 seconds and the permeability of this solution is 94.7 percent. From these values is the superiority of the invention manufactured product can be seen.

Various freeze-dried cyclocytidine preparations with or without additives were produced under the conditions of the formation of insoluble substances, ie at a plate ^{temperature of about 20 to 30 °} C., and the permeability and the rate of dissolution of the preparations were determined. The results are summarized in Table I. The permeability means the percentage permeability of a 10 weight percent aqueous solution of the freeze-dried preparation at a wavelength of 400 μm (the percentage permeability

permeability of distilled water as the standard is assumed to be 100). The dissolution rate is the time in seconds which is necessary to completely dissolve the preparation if water is added to the dry preparation in 10 times the amount by weight and the mixture is gently stirred. the end Table I are the effects of those used in the present invention

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Additions visible.

The examples illustrate the invention. Parts, percentages and proportions relate to weight, provided nothing else is indicated.

example 1

50 g of cyclocytidine and 1 g of sorbitol are dissolved in distilled water for injection purposes. 500 ml of an aqueous solution of these compounds are prepared. The solution is filtered through a membrane filter with a pore size of 0.22 microns under sterile conditions and filled in an amount of 5 ml in a sterile manner into 20 ml glass vessels. The glass vessels are placed on a tray and placed on a plate in a freeze-drying chamber. Devices for determining the electrical resistance and the temperature are placed in separate glass vessels. The temperature of the system will gradually decrease to around -40 ° C. At a temperature of the frozen solution obtained around -40 ° C, the electrical resistance assumes the value infinite. The system is gradually evacuated and at the same time the temperature of the plate is set to about 30 ^° C. by heating the plate. As the preparation continues to dry, the temperature of the plate is gradually reduced to about 20 ^° C. As soon as the temperature of the freeze-dried preparation has reached the same temperature as the plate, the preparation is kept at this temperature for 1 hour until the freeze-drying is complete. ·

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Example 2

50 g of cyclocytidine and 1 g of glucose are dissolved in distilled water of injection quality to produce 500 ml of an aqueous solution. The aqueous solution is filtered through a membrane filter under sterile conditions and filled into sterilized glass vessels in an amount of 5 ml each time. The solution according to Example 1 is then freeze-dried ^{at a plate temperature of 30 ° C.}

Example "3

In injectable quality distilled water, 50 g Cyclocytidine and 1 g of polyvinylpyrrolidone dissolved for the preparation of 500 ral of an aqueous solution. The solution is given by a Membrane filters are filtered under sterile conditions, and 5 ml of the solution are poured into sterilized glass vessels and freeze-dried at a plate temperature of 30 ° C according to Example 1.

Example 4

50 g of cyclocytidine and 1.5 g of sorbitol are dissolved in distilled water of injection quality to produce 500 ml of an aqueous solution. The solution is filtered through a membrane filter under sterile conditions, and in each case 5 ml v / earth is filled into sterilized glass vessels. The solution is freeze-dried according to example 1 at a plate temperature of ⁰ C 3Q.

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Example 5

50 g of cyclocytidine and 2 g of sorbitol are dissolved in distilled water of injection quality to produce 500 ml of an aqueous solution. The solution is filtered through a membrane filter under sterile conditions, and each 5 ml are filled into sterilized glass vials and freeze-dried according to example 1 at a plate temperature of 3O ⁰ C.

Example 6

In injection quality distilled water, 50 g of cyclocytidine and 0.02 g of HCO-50 are dissolved to make 500 ml of an aqueous solution. The solution is filtered through a membrane filter under sterile conditions, and in each case 5 ml are filled into sterilized glass vessels and freeze-dried ^{according to Example 1 at a plate temperature of 30 ° C.}

Example 7

50 g of cyclocytidine and 2 g of polyvinylpyrrolidone are dissolved in distilled water of injection quality to produce 500 μl of an aqueous solution. The solution is filtered through a membrane filter under sterile conditions, and in each case 5 ml are filled into sterilized glass vessels and freeze-dried according to Example 1 at a plate temperature of 30 ^{° C.}

Example 8

In injection-quality distilled water, 50 g Cyclocytidine and 2 g of glucose to make 500 ml of one

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dissolved aqueous solution. The solution is filtered through a membrane filter filtered under sterile conditions, and each 5 τηΐ are Filled into sterilized glass vessels and freeze-dried according to Example 1 at a plate temperature of 30 ° C.

Example 9

50 g of cyclocytidine and 2 g of xylitol are dissolved in distilled water of injection quality to produce 500 ml of an aqueous solution. The solution is filtered through a membrane filter under sterile conditions, and in each case 5 ml of the solution are filled into sterilized glass vessels and freeze-dried ^{according to Example 1 at a plate temperature of 30 ° C.} .

Example 10

According to Example 1, various freeze-dried preparations are made using the additives and amounts given in Table I. The results are also in Table I summarized.

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Table I. Passage Resolution example additive Lot, sweetness, speed No. % % eligibility, sec. 89.6 9.5 10 Sorbitol 1 94.7 7.4 1 2 98.3 14.0 4th 3 97.9 17.9 5 4th 85.2 20.2 10 5 82.3 24.6 10 6th 90.1 17.8 10 Mannitol 1 92.0 20.1 10 4th 93.8 16.4 10 Polyvinyl 0.4 94.4 - 10 pyrrolidone (PVP) 1 95.3 - 3 2 95.0 11.5 7th 4th 94.3 7.9 10 Xylitol 1 96.0 11.8 . 9 4th 92.0 26.6 10 Lactose 1 95.0 30.1 10 4th 94.1 32.0 10 glucose 1 94.9 35.0 2 2 96.3 40.0 8th 4th 90.3 19.4 10 Fructose 1 91.0 21.4 10 4th 87.5 24.3 10 5 83.3 26.6 10 6th 84.8 11.5 10 Dextran 250 1 85.1 14.8 . 10 4th 81.3 17.1 10 5 91.3 9.8 10 Dextran 20 1 79.0 20.2 10 4th 96.5 7.4 6th HCO-50 0.04 86.4 14.9 10 0.004 91.6 15.3 10 CMC-Na 1 Jf / -V Sorbitol 1 7.9 10 HCO-50 0.04 • 94.1 29.4 10 PVP
glucose 1
1 80.3 45.8 comparison No addition 0

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Remarks;

1. Amount added: percent by weight, based on cyclocytidine

2. HCO-50:

3. Dextran 250:

Dextran 20:

Ethylene oxide adduct of hydrogenated castor oil

Dextran with an average molecular weight of 250,000.

Dextran with an average molecular weight of 20,000.

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Claims (7)

Claims 1. Process for the preparation of freeze-dried 2,2'-anhydro-1-ß-D-arabinofuranosylcytosine preparations, characterized in that a 2,2'-anhydro-1-ß-D ~ aribinofuranosylcytosin and (1) a saccharide, (2) polyvinylpyrrolidone, (3) sodium-carboxymethyl cellulose or (4) containing a nonionic wetting agent or a mixture of at least two of these compounds freeze-dried aqueous solution containing components (1), (2) and (3) in an amount of 0.1 to 5 percent by weight each and the component (4) in an amount of 0.01 to 0.05% by weight based on the cytosine compound. 2. The method according to claim 1, characterized in that one used as saccharide fructose, sorbitol, xylitol, mannitol, glucose, lactose or dextran. 3. The method according to claim 1, characterized in that the non-ionic network center], a sorbitan fatty acid ester · - Ethylene oxide adduct, an ethylene oxide adduct of hardened castor oil or an ethylene oxide-propylene oxide block copolymer is used, 4. The method according to claim 1, characterized in that the freeze-drying is carried out at a plate temperature of the freeze-drying chamber of 20 to 30 ^° C. " t 409843/1105 5. The method according to claim 1, characterized in that an aqueous solution is used, the 50 parts by weight of Cytosine compound and 1 to 2.5 parts by weight sorbitol or 0.15 to 2.5 parts by weight of polyvinylpyrrolidone or 0.05 to 1 part by weight of sodium carboxymethyl cellulose or 0.005 to 0.025 part by weight of an ethylene oxide adduct of cured Contains castor oil in sterilized distilled water. 6. The method according to claim 1, characterized in that an aqueous solution is used which contains 50 parts by weight of the cytosine compound, 0.15 to 2 parts by weight of polyvinylpyrrolidone and contains 0.5 to 2 parts by weight of glucose in sterilized distilled water. 7. Injectable preparation, consisting of an aqueous solution of the freeze-dried preparation produced according to claims 1 to 6. 409849/1105