JP2003095955A - Stabilized solid composition of sodium hyaluronate - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a composition of sodium hyaluronate having enhanced stability to heat. SOLUTION: This stabilized composition of the sodium hyaluronate comprises the sodium hyaluronate as an active ingredient and saccharides added thereto.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a heat-stable solid composition of sodium hyaluronate. Also,
This is a method for suppressing a decrease in the intrinsic viscosity value of sodium hyaluronate when an aqueous solution of sodium hyaluronate is freeze-dried.

[0002]

2. Description of the Related Art Hyaluronic acid, which is a constituent of sodium hyaluronate, is an acidic polysaccharide that is commonly found in the umbilical cord, synovial fluid, vitreous body, and skin, and its solution is colorless, odorless, and highly viscous. . Hyaluronic acid also plays an important role in maintaining skin elasticity and homeostasis due to its high water retention capacity. During the regeneration process of skin tissue, the local concentration of hyaluronic acid is increased, giving an optimal state for activation, migration and proliferation of cells involved in tissue regeneration.

It is known that hyaluronic acid or its salts gradually decrease in molecular weight during storage at room temperature, and a patent for a formulation which suppresses the decrease in molecular weight and improves stability has been filed (Japanese Patent Application Laid-Open No. Hei 10 (1999) -242945). 8-104642, JP-A-9-176020).
In Japanese Patent Application Laid-Open No. 8-104642, stability is improved by adding citric acid or citrate to a solution of sodium hyaluronate in a phosphate buffer solution, and in Japanese Patent Application Laid-Open No. 9-176020, glycerin is further added. Is added to improve stability.
It has also been reported that mannitol and polyphenol, which are hydroxy radical scavengers, are effective as stabilizers of hyaluronic acid. However, all of these are stable as an aqueous solution, but when made into a solid, the molecular weight is remarkably reduced at room temperature, and it is necessary to store them in a refrigerator or a freezer. Therefore, there was no hyaluronic acid solid composition stable at room temperature.

[0004]

DISCLOSURE OF THE INVENTION The present invention relates to the stabilization of sodium hyaluronate as a solid. The details will be described below.

The origin of the sodium hyaluronate used in the present invention is not limited, and a chicken cob, a microorganism, or any other origin can be used. Further, the molecular weight is not specified, and any available molecular weight can be used, but the molecular weight is generally about 100,000 to 5,000,000.
The preferred molecular weight of hyaluronic acid is 400,000 to 2.4 million.

In the present invention, the content of sodium hyaluronate as a solid is 1% to 50%, preferably 2%.
It is 5% to 50%.

The present invention can suppress the lowering of the molecular weight of sodium hyaluronate by adding saccharides. The saccharides used in the present invention are glucose, galactose, mannose, fructose, sorbose and arabinose. , Ribose, xylose, deoxyglucose, fucose, rhamnose, galaheptulose, mannoheptulose, trehalose, sophorose, nigerose, laminaribiose, maltose, cellobiose, isomaltose, gentiobiose, sucrose, lactose, melibiose, raffinose, It may be selected from melezitose, mannitol, glucosamine, galactosamine and inositol. A particularly preferred sugar is
Maltose, sucrose, lactose and mannitol. These saccharides may be used alone or in combination of two or more. These sugars are physiologically acceptable substances for humans and animals. It is generally preferable that these contents are equal to or more than the same weight of sodium hyaluronate. In the present invention, the content of saccharide as a solid is 50% to 9%.
9% is preferable.

The sodium hyaluronate solid composition of the present invention is generally produced by a freeze-drying method. Freeze-drying is used in serum, pharmaceuticals, instant foods, etc., and is a method of rapidly freezing a heat-labile substance to maintain a high vacuum and removing water by sublimation to dry it. In the field of pharmaceuticals, it is commonly used for the production of injectables such as antibiotics that are sensitive to moisture and heat.

In the freeze-drying method, by adjusting the cooling rate until freezing the solution, the freezing temperature / time, the degree of vacuum, the temporary drying (sublimation) temperature / time, and the secondary drying (drying) temperature / time, The dry state can be changed. In the present invention, sodium hyaluronate and saccharides are preferably dissolved in an aqueous medium, and the freezing temperature is −40 ° C. and the freezing time is 2 hours. In this case, the aqueous medium is water in principle, but alcohol or other water-miscible organic solvent may be added if necessary. However, it is not necessary to significantly change the freezing conditions. Then, using a vacuum degree of about 0.1 mTorr, primary drying is performed at about 10 ° C for 36 to 39 hours, and secondary drying is performed at about 40 ° C for 3 hours.
Freeze-drying is completed by carrying out ~ 4 hours.

[0010]

EFFECTS OF THE INVENTION The effect of the present invention is to provide a composition containing sodium hyaluronate, which has a very low molecular weight reduction of sodium hyaluronate and can be stored at room temperature for a long period of time. Also, by adding these to an aqueous solution of sodium hyaluronate,
An effect of suppressing a decrease in the intrinsic viscosity value of hyaluronic acid in the freeze-drying process can be obtained.

Therefore, the present invention provides a composition containing sodium hyaluronate, which has excellent stability and convenience in use, which cannot be achieved by conventional products. By supplying it in a more stable solid state, it can be used as a solution at the time of use for various skin diseases and wounds, as well as in sheet form, sponge form,
It can be used in a solid or solid state such as powder, ointment or cream. A therapeutic agent with various forms and dosage forms of sodium hyaluronate can be expected.

A specific application example in the solid state is a state in which the water absorption characteristic of a substance is increased by freeze-drying, so that the exudate from the wound part has a high absorbability, and the part that has absorbed the exudate is gelled. It exerts the effect of a cover against the external air contact of the wound surface, and as a result, can promote healing while protecting the original repair process of the skin.

For dry skin diseases, an appropriate amount of water may be added to make a viscous state and then applied. At this time, an antibacterial agent or an antipruritic agent can be mixed and applied.
For moist skin diseases such as eczema and dermatitis, it can be applied as it is or by admixing a steroid agent or an antipruritic agent. For a skin defect site, a granulation promoter or an epithelialization promoter can be mixed and used as an adjunct drug. A decubitus wound can be treated by treating the decubitus wound with a sponge due to the granulation effect of sodium hyaluronate. At this time, a granulation growth factor or the like can be mixed and used.

Further, by using the solidified product by freeze-drying, it becomes possible to prepare a dosage form such as an ointment, a cream, a powder for external use, and various dosage forms can be prepared for various skin diseases. It will be possible. Another possible use is to freeze-dry with antibacterial agents and anti-inflammatory agents to adapt to the wound.

Other advantages include simplification of transportation means and ease of storage. What was originally stored in a cold place (frozen) during transportation is now stable at room temperature, so equipment such as a cool box is not required and equipment costs can be reduced.

Examples of the present invention will be shown below together with experimental examples and comparative examples to more specifically describe the present invention.

[0017]

Example 1 Prescription sodium hyaluronate 1 g (average molecular weight 1 million) Maltose 1 g Water for injection 98 g Total amount 100 g

Preparation Method Water for injection was added to and dissolved in maltose, and then sodium hyaluronate was added and dissolved with stirring to obtain a uniform solution. This solution was filled in a glass vial and freeze-dried. It was possible to prepare a powder for external use by adding talc to 2 g of the freeze-dried product to make 100 g and mixing them uniformly.

Freeze-Drying Method A glass vial filled with the solution was placed in a freeze-drying machine, cooled to −40 ° C. for 2 hours, and then vacuum evacuation was started.
Then set the shelf temperature of the freeze dryer to 10 ° C and
After primary drying for 3 hours and secondary drying for 3 to 4 hours at a shelf temperature of 40 ° C., sealing was performed. Freeze-drying was performed thereafter under the same conditions. When the freeze-dried product was observed with an electron microscope, no crystals of the added saccharide were observed.

Example 2 Formulation Sodium hyaluronate 1 g (average molecular weight 1 million) Mannitol 1 g Water for injection 98 g Total amount 100 g

Preparation Method Water for injection was added to mannitol and dissolved, and then sodium hyaluronate was added and dissolved by stirring to obtain a uniform solution.
This solution was filled in a glass vial and freeze-dried.

Example 3 Prescription sodium hyaluronate 1 g (average molecular weight 1 million) Sucrose 1 g Water for injection 98 g Total 100 g

Preparation Method Water for injection was added to sucrose and dissolved, and then sodium hyaluronate was added and dissolved by stirring to obtain a uniform solution. Fill this solution into a glass vial,
Lyophilized.

Example 4 Prescription sodium hyaluronate 1 g (average molecular weight 1 million) Lactose 1 g Water for injection 98 g Total amount 100 g

Preparation Method Water for injection was added to lactose and dissolved, and then sodium hyaluronate was added and dissolved by stirring to obtain a uniform solution. Fill this solution into a glass vial,
Lyophilized.

Example 5 Formulation Sodium hyaluronate 1 g (average molecular weight 2,000,000) Maltose 1 g Water for injection 98 g Total amount 100 g

Preparation Method Water for injection was added to maltose and dissolved, and then sodium hyaluronate was added and dissolved by stirring to obtain a uniform solution. This solution was filled in a glass vial and freeze-dried.

Example 6 Prescription sodium hyaluronate 1 g (average molecular weight 2,000,000) Mannitol 1 g Water for injection 98 g Total amount 100 g

Preparation Method Water for injection was added to mannitol and dissolved, and then sodium hyaluronate was added and dissolved by stirring to obtain a uniform solution.
This solution was filled in a glass vial and freeze-dried.

Example 7 Prescription sodium hyaluronate 1 g (average molecular weight 2,000,000) Sucrose 1 g Water for injection 98 g Total amount 100 g

Preparation Method Water for injection was added to sucrose and dissolved, and then sodium hyaluronate was added and dissolved by stirring to obtain a uniform solution. This solution was filled in a glass vial and freeze-dried.

[0032]

Comparative Example Comparative Example 1 Prescription sodium hyaluronate 1 g (average molecular weight 1 million) Maltose 1 g Water for injection 98 g Total amount 100 g

Preparation Method Water for injection was added to maltose and dissolved, then sodium hyaluronate was added and dissolved by stirring to obtain a uniform solution.

Comparative Example 2 Prescription sodium hyaluronate 1 g (average molecular weight 1 million) Mannitol 1 g Water for injection 98 g Total amount 100 g

Preparation Method Water for injection was added to mannitol and dissolved, and then sodium hyaluronate was added and dissolved by stirring to obtain a uniform solution.

Comparative Example 3 Prescription sodium hyaluronate 1 g (average molecular weight 1 million) Sucrose 1 g Water for injection 98 g Total amount 100 g

Preparation Method Water for injection was added to sucrose and dissolved, and then sodium hyaluronate was added and dissolved by stirring to obtain a uniform solution.

Comparative Example 4 Prescription sodium hyaluronate 1 g (average molecular weight 1 million) Lactose 1 g Water for injection 98 g Total amount 100 g

Preparation Method Water for injection was added to lactose and dissolved, and then sodium hyaluronate was added and dissolved by stirring to obtain a uniform solution.

Comparative Example 5 Prescription sodium hyaluronate 1 g (average molecular weight 2,000,000) Maltose 1 g Water for injection 98 g Total amount 100 g

Preparation Method Water for injection was added to maltose and dissolved, then sodium hyaluronate was added and dissolved by stirring to obtain a uniform solution.

Comparative Example 6 Prescription sodium hyaluronate 1 g (average molecular weight 2,000,000) Mannitol 1 g Water for injection 98 g Total amount 100 g

Preparation Method Water for injection was added to mannitol and dissolved, and then sodium hyaluronate was added and dissolved by stirring to obtain a uniform solution.

Comparative Example 7 Prescription sodium hyaluronate 1 g (average molecular weight 2,000,000) Sucrose 1 g Water for injection 98 g Total amount 100 g

Preparation Method Water for injection was added to sucrose and dissolved, and then sodium hyaluronate was added and dissolved by stirring to obtain a uniform solution.

Comparative Example 8 Prescription sodium hyaluronate 1 g (average molecular weight 1 million) Water for injection 99 g Total amount 100 g

Preparation Method Sodium hyaluronate was added to water for injection and stirred to make a uniform solution.

Comparative Example 9 Prescription sodium hyaluronate 1 g (average molecular weight 1 million) Water for injection 99 g Total amount 100 g

Preparation Method Sodium hyaluronate was added to water for injection and stirred to form a uniform solution, which was filled in a glass vial,
Lyophilized.

Comparative Example 10 Prescription sodium hyaluronate 1 g (average molecular weight 2,000,000) Water for injection 99 g Total amount 100 g

Preparation Method Sodium hyaluronate was added to water for injection and stirred to give a uniform solution, which was filled in a glass vial,
Lyophilized.

[0052]

[Experimental Example] Experimental Example 1 Sodium hyaluronate having an average molecular weight of 1,000,000 and 2,000,000 was prepared in a solution and in a freeze-dried state, and the change in viscosity under a temperature load of 60 ° C for 10 days was measured by limiting viscosity. confirmed. Tables 1 and 2 show the comparison results of the intrinsic viscosity values of each example.

[0053]

[Table 1]

[0054]

[Table 2]

Hyaluronic acid tends to be less stable against temperature as the molecular weight increases. From these results, the stabilizing effect in the solid state was obtained by adding saccharides. In addition, maltose and sucrose were superior in stabilizing effect to the mannitol-added solution, which was a hydroxy radical scavenger that was reported to be effective as a stabilizer for hyaluronic acid.

Experimental Example 2 From the results of Experimental Example 1, the temperature conditions were further changed and the temperature load period was extended. Maltose and sucrose were selected as additives with respect to sodium hyaluronate (average molecular weights of 1,000,000 and 2,000,000), and the viscosity change with temperature in the freeze-dried state was confirmed by measuring the intrinsic viscosity. Table 3 shows the results of comparison immediately after production and after 3 months of temperature loading (up to 2 months for an average molecular weight of 2,000,000).
And Table 4. As a comparative control, untreated sodium hyaluronate (average molecular weight 1,000,000) and non-added lyophilized sodium hyaluronate (average molecular weight 2,000,000) were also subjected to the same temperature loading.

[0057]

[Table 3]

[0058]

[Table 4]

Experimental Example 3 Since the intrinsic viscosity was lowered by performing freeze-drying, an additive-free solution containing only sodium hyaluronate, a freeze-dried solution containing no sodium hyaluronate, maltose and sucrose were respectively added to sodium hyaluronate. The same weight added solution,
Table 5 shows the results of comparison of the freeze-dried solutions immediately after their preparation. All the solutions are 1
It was set to 00%.

[0060]

In the case of no addition, the solution was freeze-dried to 100%.
Although the decrease in the intrinsic viscosity value was observed from 100% to 75%, when it was added, almost no change was observed from 100% to 98% by freeze-drying the solution. From this, it was observed that the addition of maltose and sucrose had an inhibitory effect on the reduction of the intrinsic viscosity value during freeze-drying.

Claims (10)

[Claims] 1. One or two kinds of sodium hyaluronate
A solid composition comprising at least one saccharide. 2. The composition according to claim 1, which is obtained by freeze-drying an aqueous solution containing sodium hyaluronate and one or more kinds of sugars. 3. One or more sugars are glucose, galactose, mannose, fructose, sorbose, arabinose, ribose, xylose, deoxyglucose, fucose, rhamnose, galaheptulose, mannoheptulose, trehalose, The sophorose, nigerose, laminaribiose, maltose, cellobiose, isomaltose, gentiobiose, sucrose, lactose, melibiose, raffinose, melezitose, mannitol, glucosamine, galactosamine and inositol according to claim 1 or 2. Composition. 4. The content of sodium hyaluronate is 1% to.
Composition according to claim 1 or 2, which is 50%, preferably 25% to 50%. 5. The composition according to claim 1, wherein the saccharide content is 50% to 99%. 6. A sodium hyaluronate-containing solid composition having improved heat stability by the addition of one or more saccharides. 7. A hyaluronic acid-containing solid composition having improved stability to heat when freeze-dried from an aqueous solution containing one or more kinds of sugars. 8. A method of suppressing the decrease in the intrinsic viscosity value of sodium hyaluronate during the freeze-drying process of the solution by adding one or more saccharides to the aqueous solution of sodium hyaluronate. 9. A method for stabilizing a hyaluronic acid-containing solid composition, which comprises freeze-drying an aqueous solution containing one or more sugars. 10. One or more sugars are glucose, galactose, mannose, fructose, sorbose, arabinose, ribose, xylose, deoxyglucose, fucose, rhamnose, galaheptulose, mannoheptulose, trehalose, 10. Stabilization according to claim 9 selected from the group consisting of sophorose, nigerose, laminaribiose, laminaribiose, maltose, cellobiose, isomaltose, gentiobiose, sucrose, lactose, melibiose, raffinose, melezitose, mannitol, glucosamine, galactosamine and inositol. Method.