JPH1087499A - Sterile composition and its production - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a sterile composition comprising N<4> -behenoyl-1-β- Darabinosylcytosine and a polyoxyethylene-added oxyfatty acid glycerol ester, through sterilizing filtration. SOLUTION: This sterile composition is easily obtained by the following process: a solvent is added to (i) N<4> -behenoyl-1-β-Darabinosylcytosine and (ii) a polyoxyethylene-added oxyfatty acid glycerol ester to prepare a solution which, in turn, is subjected to sterilizing filtration followed by distilling the solvent off to afford residuals; while a mixture of (iii) dehydrocholic acid and (iv) auxiliary additive(s) composed of saccharide, common salt, thiosulfuric acid compound and/or benzoic acid compound, and if needed, a pH adjustor is dissolved in sterile water and then subjected to sterilizing filtration to afford a solution; and the above residuals are then dissolved in this solution, and the resultant solution is subjected to sterilizing filtration again.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

[0001] The present invention relates to a sterile composition and a method for producing the same. More specifically, the present invention relates to N
⁴ -behenoyl-1-β-D-arabinofuranosylcytosine (generic name enocitabin
The present invention relates to a sterile pharmaceutical composition comprising e)) and a method for producing the same.

[0002]

BACKGROUND OF THE INVENTION N ⁴ - N ⁴ belonging to the acyl -1-β-D- arabinosylcytosine derivatives - behenoyl -1-β-
D-arabinofuranosylcytosine is an antineoplastic agent having an excellent antitumor effect, and has been discovered by the present inventors (Japanese Patent Application Laid-Open No. 50-18482, Japanese Patent Publication No. 53-1983).
No. 5678, Patent No. 0928188). Since this compound is a derivative having a behenoyl group, it is insoluble in water and oil and has a problem in bioavailability. (JP-A-53-26317, JP-B-55-263).
26123, Japanese Patent No. 1035215). However, N ⁴ - of the acyl -1-beta-D-araC derivatives, in particular N ⁴ - behenoyl -1-beta-D-
There have been various formulation technology difficulties in producing formulations of arabinofuranosyl cytosine.

[0003]

In general, it is natural that strict sterilization is required for medicines, especially for injections. Therefore, in the case of injections, in the final step, sterile-filtered drugs are aseptically sterilized. Enclosure in containers has been performed. The injectable preparation of N ⁴ -behenoyl-1-β-D-arabinofuranosylcytosine is extremely difficult to pass through an aseptic filter for sterilization in the final step of preparation. It was necessary to remove bacteria by a method, and the production process was complicated. Also, for this purpose, the individual components in the formulation are dissolved in a solvent in which each component can be dissolved, and then individually filtered through a sterile filter,
Aseptic substances of each component were obtained and mixed to form a preparation, which was similarly difficult to pass through a sterile filter.

[0004]

In particular, when N ⁴ -behenoyl-1-β-D-arabinofuranosylcytosine is dissolved in a soluble solvent, the solution does not pass through fungi. Although the solution is easily passed through a filter, the solution obtained by heating and dissolving the solution is difficult to pass through the filter.
As a result of various studies to solve this problem,
Surprisingly, a solution containing N ⁴ -behenoyl-1-β-D-arabinosylcytosine and a glycerol ester of an oxyfatty acid to which polyoxyethylene has been added passes through a sterile filter even after being heated and dissolved. The resulting composition was passed through a sterile filter to obtain sterile N ⁴ -behenoyl-1-β-D-.
A composition containing arabinofuranosylcytosine was obtained, thereby completing the present invention.

[0005] The present invention is based on the above findings, N ⁴
A sterile composition containing -behenoyl-1-β-D-arabinofuranosylcytosine and obtained by sterile filter filtration, preferably N ⁴ -behenoyl-1-β-D
A sterile composition obtained by filtration using a sterile filter comprising a solution of arabinofuranosylcytosine and a glycerol ester of an oxy fatty acid to which polyoxyethylene has been added; and the present invention further provides (i) N ⁴ -behenoyl-
A residue obtained by filtering a solution containing 1-β-D-arabinofuranosylcytosine and (ii) a glycerin ester of oxyfatty acid to which polyoxyethylene has been added through a sterile filter and removing the solvent, and A sterile composition comprising (iii) dehydrocholic acid and (iv) an auxiliary additive consisting of sugar or salt, sulfuric acid, and benzoic acid, and if necessary, a required amount of a pH adjuster, and a process for producing the same.

That is, the present invention relates to N ⁴ -behenoyl-1
An object of the present invention is to provide a sterile composition containing -β-D-arabinofuranosylcytosine and obtained by filtration through a sterile filter. Furthermore, the present invention provides (i) N ⁴ -behenoyl-
A solution containing 1-β-D-arabinofuranosylcytosine and (ii) a glycerol ester of oxyfatty acid to which polyoxyethylene has been added is filtered through a sterile filter, and the solvent is removed to obtain a residue. And a sterile composition comprising (iii) dehydrocholic acid and (iv) saccharides or salt, thiosulfates, and benzoic acids.

The present invention further relates to (i) N ⁴ -behenoyl-1-β-D-arabinofuranosylcytosine, and (i)
i) A solution obtained by adding a solvent to a glycerol ester of an oxy fatty acid to which polyoxyethylene has been added and filtering the solution through a sterile filter, removing the solvent to obtain a residue,
(iii) dehydrocholic acid and (iv) sugar or salt,
Auxiliary additives consisting of thiosulfates and benzoic acids were added,
Dissolve the mixture of (iii) and (iv) by adding sterile water,
N ⁴ for the solution with a solution obtained by filtration with a sterile for filter, characterized in that said (i) solution of sterile for filter filtration residue (ii) filtering to sterile for filter - It is intended to provide a method for producing a sterile composition of behenoyl-1-β-D-arabinofuranosylcytosine.

In the present invention, N ⁴ -behenoyl-1-β
-D-arabinofuranosylcytosine is disclosed in
It can be synthesized by the method disclosed in JP-A-8482.

The glycerol ester of oxyfatty acid to which polyoxyethylene has been added is preferably one in which the polyoxyethylene portion has an average of 10 to 200 moles of oxyethylene polymerized, and particularly preferably one in which an average of 40 to 100 moles has been polymerized. In addition, the number of carbon atoms of the oxy fatty acid residue is 1
Those having 0 to 20 are preferred, and oxystearic acid is particularly preferred. Since hydrogenated castor oil is mainly composed of triglyceride of oxystearic acid, the average degree of polymerization is 40 to 40%.
Polyoxyethylene hydrogenated castor oil to which 100 mol of polyoxyethylene has been added may be mentioned, and a particularly preferred example is polyoxyethylene hydrogenated castor oil 60 (HCO-
60).

The amount of the glycerol ester of oxy fatty acid to which polyoxyethylene is added is 0.1 to 20 parts by weight based on 1 part by weight of N ⁴ -behenoyl-1-β-D-arabinofuranosylcytosine. Is preferred. 0.
If the amount is less than 1 part by weight, no effect is exhibited, and if the amount is more than 20 parts by weight, no special effect is exhibited, and the concentration of N ⁴ -behenoyl-1-β-D-arabinofuranosylcytosine is reduced. Not economic.

The solvent used for preparing the solution of N ⁴ -behenoyl-1-β-D-arabinofuranosylcytosine and the glycerol ester of oxyfatty acid to which polyoxyethylene has been added is a solvent capable of dissolving both components. It is not limited at all. Examples of the solvent for both components include ketones such as acetone and methyl ethyl ketone, esters such as ethyl acetate and butyl acetate, chain ethers such as ethyl ether and methyl ethyl ether, and cyclic ethers such as tetrahydrofuran, tetrahydropyran and dioxane. Amides such as dimethylacetone amide, dimethylformamide and diethylacetamide, sulfoxides such as dimethyl sulfoxide,
Examples include alcohols such as methanol, ethanol, n-propanol and isopropanol, bases such as pyridine and triethanolamine, and acids such as formic acid and acetic acid.

Among these solvents, ketones, esters,
The aprotic solvents such as chain ethers, cyclic ethers, amides, and sulfoxides are those described above for N ⁴ -behenoyl-1.
Preferred is that it does not degrade -β-D-arabinofuranosylcytosine. Ketones, esters, chain esters, cyclic monoethers, and alcohols are preferred because they are easily distilled off. Ketones, amides and ethanol are preferred because of their low toxicity. Ethanol is particularly preferred because of ease of distillation and low toxicity.

Next, the filter used for filtering the above solution is not particularly limited as long as it is a filter that can be sterilized, and examples thereof include a glass filter and a cellulose mixed ester filter.
These filters may be preliminarily sterilized by an autoclave or the like. In this way, the solution obtained by aseptic filtration with a filter does not increase the temperature by performing the distillation under reduced pressure to remove the solvent,
It is preferable because it can be distilled quickly.

Further, the sterile composition of the present invention includes N ⁴
-A filter capable of preventing fungi from passing through an aqueous solution containing glycerol ester of oxyfatty acid to which behenoyl-1-β-D-arabinofuranosylcytosine and polyoxyethylene are added, dehydrocholic acid and auxiliary additives, and if necessary, a pH regulator (sterile). Obtained by filtration with a filter for use. First, N ⁴ - As the glycerol ester of behenoyl -1-beta-D-araC and the added hydroxy fatty acid polyoxyethylene, obtained by distilling off the solvent after filtration to the above sterile filter residue Compositions.

Next, the dehydrocholic acid in the present invention may be a salt thereof, and also includes those used as dehydrocholic acid and formed into a salt with a pH adjuster, for example, sodium hydroxide. The dehydrocholic acid includes N ⁴ -behenoyl-1-β-D
-0.01 to 5 parts by weight of arabinosyl cytosine
Part by weight, preferably 0.05 to 1 part by weight may be used. In addition, as an auxiliary additive, it can be added together with the above dehydrocholic acid to appropriately adjust the target composition weight. First, as saccharides, monosaccharides and disaccharides are preferable, and glucose is particularly preferable. As thiosulfates, thiosulfuric acid and its salts are preferred, alkali metal salts of thiosulfuric acid are more preferred, and sodium thiosulfate is particularly preferred.

The benzoic acid means a carboxylic acid having a carboxy group on the benzene ring, and is preferably an aromatic carboxylic acid such as benzoic acid or salicylic acid, or a salt thereof. Salts or ammonium salts are more preferred, with sodium salicylate being particularly preferred.

Since the pH of the aqueous solution of the composition varies depending on the combination of each component of the preparation of the present invention, a pH adjuster is added if necessary. As the pH adjuster, an inorganic or organic acid or an inorganic or organic alkali is used. Among them, 6 hydrochloric acid or sodium hydroxide is convenient.

The aqueous solution from which the sterile filter can be filtered is
For example the filtrate obtained by an aqueous solution containing a pH adjusting agent was filtered through a sterile for filter if necessary and dehydrocholate auxiliary additives and, N ⁴ - behenoyl -1-beta-D-arabinofuranosyl cytosine and polyoxyethylene A solution of the glycerol ester of oxyfatty acid to which is added is filtered by a sterile filter, and the obtained filtrate is added to the residue obtained by evaporation, followed by heating and dissolving.

In order to obtain the sterile composition of the present invention as a solid, the solution is filtered through the same sterile filter as described above, which does not allow fungi to pass through, and the liquid component obtained is distilled off. In this case, lyophilization is preferred. The solid composition is soluble in water.

[0020]

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail with reference to examples, but is not limited thereto. Example 1 1 g of N ⁴ -behenoyl-1-β-D-arabinofuranosylcytosine and polyoxyethylene hydrogenated castor oil 60 (H
7 g of CO-60) was added to 100 cc of ethanol, and the resulting mixture was dissolved under heating (1 atm, 80 ° C., 10 minutes).
Quenched to room temperature with cold water. Then, the obtained ethanol solution was filtered through a filter (sterile filter) having a pore size of 0.22 μm which had been sterilized in an autoclave beforehand, and ethanol was distilled off from the filtrate (0.1 atm, 60 atm).
C., 4 hours) to give about 1: 7 consisting of N ⁴ -behenoyl-1-β-D-arabinosylcytosine and HCO-60.
7.5 g of a composition having the following composition:

Example 2 95 mg of dehydrocholic acid, 5 g of glucose, 100 mg of sodium salicylate, 0.5 m of sodium thiosulfate
g and 9 mg of sodium hydroxide were added to and dissolved in 200 ml of sterile water, and the aqueous solution was filtered through a sterile filter having a pore size of 0.22 μm which had been sterilized in an autoclave beforehand, and the filtrate obtained in Example 1 was used. ⁴ -Behenoyl-1-β-D-arabinofuranosylcytosine and HCO
−60, and about 1 at 90 ° C.
Heated for 5 minutes to dissolve, then quenched with cold water to room temperature, N ⁴ -
Behenoyl-1-β-D-arabinosylcytosine, HC
About 200 ml of an aqueous solution containing O-60, dehydrocholic acid, benzoic acids, thiosulfates and saccharides was obtained.

Example 3 About 200 ml of the aqueous solution obtained in Example 2 was 0.22 m in pore size previously sterilized by an autoclave.
filtered through a filter of mu, N ⁴ - behenoyl -1-beta-
About 190 cc of a sterile aqueous solution containing D-arabinosylcytosine, HCO-60, dehydrocholic acid, benzoic acids, thiosulfates, and saccharides was obtained.

Example 4 A sterile aqueous solution containing N ⁴ -behenoyl-1-β-D-arabinosylcytosine, HCO-60, dehydrocholic acid, benzoic acids, thiosulfates and saccharides obtained in Example 3 190cc and lyophilized overnight, N ⁴ - behenoyl -1
-Β-D-arabinosylcytosine, HCO-60, dehydrocholic acid, benzoic acid, thiosulfate, and saccharide:
About 11 g of a composition containing 7: 0.095: 0.1: 0.0005: 5 was obtained.

Example 5 About 11 g of the composition containing N ⁴ -behenoyl-1-β-D-arabinosylcytosine obtained in Example 4 was added to sterile water 2
After adding 00 ml, the mixture was heated at 90 ° C. for about 15 minutes and rapidly cooled to room temperature with cold water to obtain an aqueous solution containing N ⁴ -behenoyl-1-β-D-arabinosylcytosine and the like. On the other hand, in Examples 1 to 4 N ⁴ - behenoyl -1-beta-D-arabinofuranosyl without adding the sill cytosine was obtained by the same procedure N ⁴ - behenoyl -1-beta-D-arabino 200 cc of sterile water was added to 10 g of a composition not containing silcytosine,
By the same operation, an aqueous solution containing no N ⁴ -behenoyl-1-β-D-arabinosylcytosine was obtained.

Example 6 Ten CDF ₁ mice were intraperitoneally transplanted with 1 × 10 ⁵ L1210 leukemia cells, and the next day, 5 mice were transplanted with N ⁴
An aqueous solution containing behenoyl-1-β-D-arabinosylcytosine was intraperitoneally administered at 200 mg / kg body weight of the mouse in terms of N ⁴ -behenoyl-1-β-D-arabinosylcytosine, and the remaining 5 Mice were intraperitoneally administered with an equal volume of an aqueous solution without N ⁴ -behenoyl-1-β-D-arabinosylcytosine. N ⁴ -behenoyl-1-
Although 30 days after the administration of the aqueous solution containing β-D-arabinosylcytosine, 5 out of 5 mice survived, but N ⁴
Mice to which an aqueous solution containing no -behenoyl-1- [beta] -D-arabinosylcytosine had been died 30 days later, 5 out of 5 mice died. Therefore, N ⁴ -behenoyl-1-
It was confirmed that β-D-arabinosylcytosine has an effect of extending the life of mice having L1210 leukemia cells.

Example 7 The procedure of Example 1 was repeated, except that 1.8 g of sodium chloride was used instead of glucose, and N ⁴ -behenoyl-1-β-D-arabinosylcytosine and HCO- 6
A composition of about 1: 7 consisting of 0 was obtained. This composition can be used similarly as the composition described in Examples 2 to 4.

[0027]

According to the present invention, preferably, a solution comprising N ⁴ -behenoyl-1-β-D-arabinofuranosylcytosine and a glycerol ester of an oxy fatty acid to which polyoxyethylene has been added is obtained by filtration using a sterile filter. The resulting sterile composition is obtained and the solvent is removed from the composition to give a residue, which is supplemented with dehydrocholic acid and sugars or salts, thiosulfates, benzoic acids, and, if necessary, pH. Sterile compositions are readily obtained to include the required amount of modulator.

Claims (20)

[Claims] 1. A sterile composition comprising N ⁴ -behenoyl-1-β-D-arabinofuranosylcytosine and obtained by filtration through a sterile filter. 2. The method of claim 1, wherein the sterile composition is N ⁴ -behenoyl-1.
The sterile composition according to claim 1, comprising a solution of -β-D-arabinosylcytosine and a glycerin ester of an oxy fatty acid to which polyoxyethylene has been added. (I) N ⁴ -behenoyl-1-β-D-
Arabinofuranosyl cytosine, and (ii) a glycerol ester of an oxy fatty acid to which polyoxyethylene has been added,
Is filtered through a sterile filter to remove the solvent, and to the residue obtained, an auxiliary additive consisting of (iii) dehydrocholic acid and (iv) sugars or salt, thiosulfates, and benzoates is added. A sterile composition comprising: 4. The glycerin ester of an oxyfatty acid to which polyoxyethylene is added is 0.1 to 20 parts by weight based on 1 part by weight of N ⁴ -behenoyl-1-β-D-arabinofuranosylcytosine. 4. The sterile composition according to 2 or 3. 5. The glycerol ester of an oxy fatty acid to which polyoxyethylene is added has an average degree of polymerization of 10 to 200.
10 to 20 carbon atoms with moles of polyoxyethylene added
The sterile composition according to claim 4, which is a glycerin ester of an oxy fatty acid. 6. The glycerol ester of an oxy fatty acid to which polyoxyethylene has been added has an average degree of polymerization of 40 to 100.
5. The sterile composition according to claim 4, which is a triglyceride of oxystearic acid to which moles of polyoxyethylene are added or hydrogenated castor oil to which polyoxyethylene having an average degree of polymerization of 40 to 100 moles is added. 7. The sterile composition according to claim 3, wherein the saccharide is a monosaccharide or a disaccharide. 8. The sterile composition according to claim 7, wherein the monosaccharide is glucose. 9. The sterile composition according to claim 3, wherein the thiosulfate is an alkali metal salt. 10. The sterile composition according to claim 3, wherein the benzoic acids are benzoic acid, salicylic acid, or a salt thereof. 11. The sterile composition according to claim 10, wherein the salt of salicylic acid is sodium salicylate. 12. The sterile composition according to claim 3, wherein the dehydrocholic acid and auxiliary additives are filtered through a sterile filter. 13. The sterile composition according to claim 3, wherein the sterile composition is obtained by filtering a final product through a sterile filter. 14. A sterile composition comprising a filtrate obtained by filtering an aqueous solution containing dehydrocholic acid and auxiliary additives and, if necessary, a pH adjuster through a sterile filter, to obtain a filtrate.
^A residue obtained by evaporating a filtrate obtained by filtering a solution of ⁴ -behenoyl-1-β-D-arabinofuranosylcytosine and a glycerol ester of an oxy fatty acid to which polyoxyethylene has been added through a sterile filter is evaporated. The sterile composition according to claim 3, wherein an aqueous solution obtained by heating and dissolving the product is filtered through a sterile filter. 15. The sterile composition according to claim 1, wherein the composition is an aqueous solution or a lyophilized product. (I) N ⁴ -behenoyl-1-β-D
Arabinofuranosylcytosine and (ii) a glycerol ester of oxyfatty acid to which polyoxyethylene has been added, dissolved in a solvent, filtered through a sterile filter, and the solvent was removed to obtain a residue; (iii) The mixture of (iii) and (iv) to which dehydrocholic acid and (iv) saccharide or salt, thiosulfates and benzoic acids are added, is dissolved by adding sterile water to the mixture. A solution obtained by dissolving the filtration residue of the sterile filter (i) and (ii) in a solution obtained by filtration with a sterile filter, and filtering the solution through a sterile filter.
^A method for producing a sterile composition of ⁴ -behenoyl-1-β-D-arabinofuranosylcytosine. 17. The method according to claim 1, wherein the dissolution is heating.
6. The production method according to 6. 18. The method according to claim 16, wherein the solvent is ethanol. 19. A filtrate obtained by dissolving N ⁴ -behenoyl-1-β-D-arabinofuranosylcytosine in ethanol and filtering through a sterile filter, evaporating ethanol, and obtaining a residue. And dehydrocholic acid, and sugars, salt, thiosulfates, a solution consisting of water and an auxiliary additive consisting of benzoic acids, preferably dissolved by a filtrate obtained by filtration with a sterile filter, more preferably dissolved. A method for producing a sterile composition of N ⁴ -behenoyl-1-β-D-arabinofuranosylcytosine, which comprises filtering through a sterile filter and freeze-drying as necessary. 20. The method according to claim 1, wherein the dissolution is heating.
9. The production method according to 9.