JP2000336041A - Urinastatin-containing aqueous preparation characterized in containing propylene glycol - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a urinastatin-containing aqueous composition having excellent heat stability, and useful for treating respiratory diseases, allergic diseases or the like by compounding propylene glycol. SOLUTION: This composition contains propylene glycol (preferably, an official drug listed in the Japanese Pharmacopoeia; the amount of addition is 0.05-4 w/v%). The pH is preferably 5.0-7.0. The urinastatin(UTI) is purified by anion exchanger treatment or the like, and it has preferably a molecular weight of 67,000, an isoelectric point of 2-3 and a specific activity of about 1,000-4,000 unit/mg-protein. It is prepared by dissolving propylene glycol, a pH controller, a chelating agent and the like in purified water for injection, a UTI stock solution is added to the solution and, they are mixed. Subsequently, the pH of the solution is adjusted, and the volume is adjusted with purified water followed by sterilization treatment.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to an aqueous preparation containing ulinastatin.

[0002]

2. Description of the Related Art Urinastatin (hereinafter abbreviated as UTI) is a human urinary trypsin inhibitor having a molecular weight of about 6%.
It is a glycoprotein of 7000, which is known to inhibit not only proteases such as trypsin, α-chymotrypsin, and esterase, but also sugar / lipolytic enzymes such as hyaluronidase and lipase. Clinically, it is used for the treatment of acute pancreatitis, chronic recurrent pancreatitis, acute circulatory failure and the like.

By the way, UTI has a disadvantage that its activity is reduced when it is kept in an aqueous solution state. In order to overcome such disadvantages, UTI is currently marketed as a lyophilized preparation, and is administered by injection in water for injection at the time of use. In the production process of UTI preparations, heat treatment is generally performed for the purpose of removing bacteria, removing viruses, and the like. For this reason, in order to make UTI into an aqueous preparation, it is required to improve the thermal stability of UTI activity in an aqueous solution.

[0004] Various methods have been proposed for preventing a decrease in activity when UTI is stored for a long time as an aqueous preparation in which UTI is dissolved in water. JP-A-6-25011 discloses a UTI-containing liquid preparation characterized by having a low ionic strength and a pH of 7 or less. This shows that when this formulation was stored at 60 ° C. for 7 days, formation of a polymer by aggregation of UTI was suppressed. However, under the same conditions, the present inventors
When the thermal stability of I activity was examined, the activity was 1 /
3 or less. Although the proposed aqueous UTI formulation is excellent in inhibiting UTI aggregation, it shows that the thermal stability of UTI activity is not yet sufficient.

JP-A-6-135852 discloses that UTI contains about 50 units / mL to 100,000 units / mL, and glucose, maltose, xylitol, D-sorbitol, and at least one selected from sodium chloride and potassium chloride. It contains at least one selected from D-mannitol, L-aspartic acid, L-glutamic acid, L-arginine, L-lysine and L-histidine, and has a pH of 3.5.
A room temperature stable UTI injection that is ~ 5.5 is disclosed. However, there is no description of propylene glycol as an additive, and there is no description that propylene glycol improves the thermal stability of UTI activity.

Further, Japanese Patent Application Laid-Open No. 7-278015 discloses a UTI-containing liquid preparation characterized by containing a sugar alcohol. In this preparation, the sugar alcohol means a polyhydroxyalkane obtained by reducing aldose or ketose and the like, for example, mannitol, sorbitol, xylitol, glycerol and the like,
Among them, mannitol is particularly preferable. It discloses that this formulation can maintain a UTI activity during long-term storage at 25 ° C. and can provide a stable liquid formulation in which a polymer and a low-molecular-weight degradation product are hardly generated. However, there is no description about the thermal stability of UTI activity. As shown in Table 2 of Example 2, there is no description that the highest thermal stability of UTI activity is obtained when propylene glycol is added to the UTI aqueous preparation.

[0007]

DISCLOSURE OF THE INVENTION The present invention has been made to solve the problems of UTI aqueous preparations, and an object of the present invention is to provide a novel UTI-containing aqueous preparation having excellent heat stability of UTI activity. Is to do.

[0008]

DISCLOSURE OF THE INVENTION The present invention provides a UTI-containing aqueous preparation excellent in heat stability of UTI activity, comprising:
A UTI-containing aqueous preparation characterized by containing propylene glycol in order to enhance the thermal stability of the activity.
The present inventors have conducted intensive studies in order to obtain a UTI-containing aqueous preparation having excellent thermal stability of UTI activity. As a result, the addition of propylene glycol to the UTI-containing aqueous preparation has a higher UTI activity than the conventional UTI-containing aqueous preparation. The present inventors have found that an aqueous preparation having excellent heat stability can be prepared and completed the present invention.

The UTI-containing aqueous preparation containing propylene glycol of the present invention is caused by respiratory diseases such as acute pancreatitis, chronic recurrent pancreatitis, acute circulatory failure, emphysema, pneumonia and pulmonary fibrosis, increased amylase activity or lipase activity. Applicable to the treatment of diseases such as diseases, allergic diseases, rheumatoid arthritis, rotavirus infection, corneal damage, corneal ulcer, dry eye, keratoconjunctival inflammation, etc. or anti-inflammatory agents, antipruritics, dry skin prevention, skin keratinization Used as a skin protectant against skin diseases, skin aging prevention, atopic dermatitis and the like.

[0010]

DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention will be described below in detail.
UTI used in the present invention is derived from urine, cell culture,
It is not limited to any derived from genetic engineering. Further, it may be purified to the extent that it can be applied as a pharmaceutical. The UTI used in the present invention also includes those in which a part of the amino acid sequence has been deleted, substituted or added by a technique such as genetic engineering. UTI purification methods include anion exchanger treatment,
Ultrafiltration, affinity chromatography, treatment with an inorganic adsorbent, salting out, isoelectric point precipitation, chitosan treatment, insoluble trypsin treatment and the like are known. Further, a metal chelate resin treatment, a hydrophobic carrier treatment, a heat treatment and the like may be combined. The UTI thus obtained has a molecular weight of about 60,000 to 70,000 (preferably 67000), an isoelectric point of about 2 to 3,
The specific activity is about 1000-4000 units / mg protein. One UTI unit is the amount of UTI that acts on 2 μg of trypsin and inhibits the enzyme activity of 1 μg of trypsin. The UTI content of the UTI-containing aqueous preparation of the present invention is not particularly limited as long as the target disease is cured, but is usually 1 to 100.
000 units / mL, preferably 10 to 50,000 units /
mL, especially when used as eye drops 10 to 100
0 units / mL.

The propylene glycol used in the present invention is not particularly limited, but is preferably a product of the Japanese Pharmacopoeia. Such propylene glycol is sold under trade names such as propylene glycol (Asahi Glass Co., Ltd.) and ADEKA propylene glycol (Asahi Denka Kogyo Co., Ltd.) and can be easily obtained. The amount of propylene glycol added is usually 0.05
Used at ~ 4 w / v%. In the case of a preparation such as an injection or an eye drop whose osmotic pressure is preferably near isotonic, the amount of propylene glycol used is preferably 0.05 to 3 w /.
v%. Propylene glycol content of 0.05
In the case of ４4 w / v%, a UTI-containing aqueous preparation having particularly high thermal stability of UTI activity can be obtained, which is preferable.

In preparing the UTI-containing aqueous preparation of the present invention, saccharides such as xylitol, mannitol, sorbitol, glucose, glycerin, sodium chloride,
Potassium chloride, tonicity agent such as boric acid, acetic acid, phosphoric acid, boric acid, citric acid, tartaric acid, lactic acid, aspartic acid, histidine, arginine, lysine, glycine, glutamic acid, ε-aminocaproic acid and their pharmaceutically Tolerable salts, pH adjusters such as sodium hydroxide and hydrochloric acid, benzalkonium chloride, benzethonium chloride, inverted soaps such as chlorhexidine gluconate, parabens such as methyl parahydroxybenzoate and propyl parahydroxybenzoate Sorbic acid and its pharmaceutically acceptable salts, preservatives such as benzyl alcohol, phenethyl alcohol or chlorobutanol, methylcellulose, hydroxypropylmethylcellulose, polyethylene glycol,
Synthetic polymers such as polyvinylpyrrolidone, polyvinyl alcohol and carboxyvinyl polymers, sodium polyacrylate, proteinaceous polymers such as gelatin, hyaluronic acid, dextran and carrageenan, polysaccharides such as sodium chondroitin sulfate, and gums such as xanthan gum Chelating agents such as EDTA or a pharmaceutically acceptable salt thereof, citric acid or a pharmaceutically acceptable salt thereof, tocopherol and its derivatives sodium nitrite, sodium sulfite, ascorbic acid And the like may be added to the UTI-containing aqueous preparation of the present invention as long as the effects of the present invention are not impaired.

The UTI-containing aqueous preparation of the present invention usually has a pH of 3
~ 7, but from the viewpoint of stability and irritation, pH5.
The pH is preferably from 0 to 7.0, and particularly preferably from 5.5 to 7.0 when used as eye drops.

The UTI-containing aqueous preparation of the present invention can be subjected to sterilization by filtration sterilization using a membrane filter, intermittent sterilization, or the like. Further, the UTI-containing aqueous preparation of the present invention can be filled in a plastic eye dropper and used as an eye drop. The UTI-containing aqueous preparation of the present invention can be filled in a plastic drop bottle and used as a liniment, lotion, or eardrop. UTI of the present invention
The contained aqueous preparation can be filled into a nasal metering sprayer and used as a nasal drop. The UTI-containing aqueous preparation of the present invention is sealed after filling into an ampoule and injected (intravenous injection, arterial injection,
Subcutaneous injection, intradermal injection, intramuscular injection, intrathecal injection, intraperitoneal injection, intraocular injection, etc.), oral solution, inhalant, and spray. Depending on the usage, it is used by filling it into a plastic drug bottle for an internal liquid, an electric nebulizer for an inhalant, and an atomizer for a spray.

Next, a method for producing the UTI-containing aqueous preparation of the present invention will be described. Various known methods are available,
For example, propylene glycol, a pH adjuster, a chelating agent, a preservative, an isotonic agent and the like are dissolved in purified water for injection. Next, the treated UTI stock solution is added and mixed. HCl or NaOH is added to adjust the target pH, and further adjusted to a predetermined volume with purified water for injection to obtain the UTI aqueous preparation of the present invention. Furthermore, the obtained UTI-containing aqueous preparation can be filtered and filled into a suitable storage container with a membrane filter, and then subjected to intermittent sterilization to prepare a prescribed dosage form.

Hereinafter, the present invention will be described in more detail with reference to Examples, Test Examples and Formulation Examples.

【Example】

Example 1 In 80 mL of water for injection, 0.5 g of ε-aminocaproic acid and 0.05 to 5 g of propylene glycol are added and dissolved. Next, add 1500000 units of UTI and mix well. After adjusting the pH to 6.0 with 1N HCl, water for injection is added to bring the total volume to 100 mL. Aside from this,
To 80 mL of water for injection, 0.5 g of ε-aminocaproic acid and 1500000 units of UTI are added and dissolved. After adjusting the pH to 6.0 with 1N HCl, water for injection is added to bring the total volume to 100 mL. 1 mL of each of the prepared propylene glycol-containing UTI aqueous solution and the propylene glycol-free UTI aqueous solution was filtered and filled into 1 mL ampoules using a membrane filter having a pore size of 0.45 μm, and then sealed.
A TI aqueous preparation and a comparative preparation containing no propylene glycol (Formulation No. 1) were prepared. UTI prepared
The aqueous formulation was stored in a thermostat at 60 ° C. for 7 days. The UTI activity before and after storage was measured, and the residual ratio of UTI activity after storage was determined. UTI activity (trypsin inhibitory activity) is N-α-
It was measured by an absorbance measurement method using benzoyl-L-arginine-p-nitroalinide as a substrate. Then, U of the formulation (formulation No. 1) in which propylene glycol was not added was added.
The degree of improvement in the UTI activity residual ratio of each formulation with respect to the TI activity residual ratio was determined by the following equation as the degree of UTI activity stability improvement, and the results are shown in Table 1. UTI activity stability improvement = (ba) × 100 / a (1) a: Formulation without addition of propylene glycol (Formulation N
o. 1) Residual ratio of UTI activity b) Each formulation containing propylene glycol (formulation N)
o. 2 to 7) Residual UTI activity From the results in Table 1, the UTI aqueous preparation containing 0.05 to 4 W / V% propylene glycol has a higher UT than the UTI aqueous preparation containing no propylene glycol.
It was shown that the thermal stability of I activity was excellent.

[0018]

[Table 1]

Example 2 0.5 g of ε-aminocaproic acid and 0.7 g of propylene glycol are added to and dissolved in 80 mL of water for injection. Next, add 1500000 units of UTI and mix well. After adjusting the pH to 6.0 with 1N HCl, water for injection is added to bring the total volume to 100 mL. 1 mL of the prepared propylene glycol-containing UTI aqueous solution was filtered and filled into a 1 mL ampoule bottle using a membrane filter having a pore size of 0.45 μm, and then sealed to prepare an aqueous formulation of a UTI-containing aqueous formulation of the present invention.

Comparative Example 1 0.7 g of 0.7 g of propylene glycol of Example 2
Was used in the same manner as in Example 2 to obtain a UTI-containing aqueous preparation for comparison.

Comparative Example 2 0.7 g of the propylene glycol of Example 2
Was used in the same manner as in Example 2 to obtain a UTI-containing aqueous preparation for comparison.

COMPARATIVE EXAMPLE 3 0.7 g of propylene glycol of Example 2 was 0.7 g
In place of xylitol, a UTI-containing aqueous preparation for comparison was obtained in the same manner as in Example 2 below.

COMPARATIVE EXAMPLE 4 0.7 g of 0.7 g of propylene glycol of Example 2
Was used in the same manner as in Example 2 to obtain a UTI-containing aqueous preparation for comparison.

The UTI-containing aqueous preparation containing ampoule obtained in Example 2 and Comparative Examples 1 to 5 was stored in a thermostat at 60 ° C. for 7 days. The UTI activity before and after storage was measured, and the residual ratio of UTI activity after storage was determined. Table 2 shows the results.
The UTI-containing aqueous preparation to which the propylene glycol of the present invention is added has a higher UT compared to the UTI-containing aqueous preparation to which a sugar alcohol is added as exemplified in JP-A-7-278015.
It was shown that the thermal stability of I activity was excellent.

[0025]

[Table 2]

Formulation Example 1 14 g of propylene glycol is added and dissolved in 100 mL of water for injection. Next, the 5000000 unit UTI
And mix well. PH was adjusted to 5.0 with 1N HCl.
After adjusting to, water for injection is added to make the total volume 1000 mL. The prepared propylene glycol-containing UTI aqueous solution was filtered and filled into ampoules using a membrane filter having a pore size of 0.45 μm, sealed, and then subjected to intermittent sterilization to prepare a UTI-containing aqueous injection of the present invention.

Ingredient UTI 50,000 units / mL Propylene glycol 1.4 w / v% 1N HCl Appropriate amount Water for injection Amount required to make the total volume 1000 ml pH 5.0

Formulation Example 2 0.22 g of methylparaben and 0.14 g of propylparaben were added to 2 g of propylene glycol, and
Dissolve while heating to 0 ° C. Here is 500m of water for injection
L, followed by 5 g of ε-aminocaproic acid and 5.9 g
And dissolve while warming to 40 ° C.
After allowing to cool to room temperature, add 150000 units of UTI and mix well. After adjusting the pH to 5.5 with 1N HCl, water for injection is added to bring the total volume to 1000 mL.
After filtering and filling the prepared propylene glycol-containing UTI aqueous solution into a plastic ophthalmic bottle using a membrane filter having a pore size of 0.45 μm, an intermittent sterilization treatment was performed.
A UTI-containing aqueous eye drop of the present invention was prepared. The prescription is shown below.

Component UTI 1500 units / mL Propylene glycol 0.2 w / v% Methyl paraben 0.026 w / v% Propyl paraben 0.014 w / v% ε-Aminocaproic acid 0.5 w / v% NaCl 0.59 w / v% 1N HCl Suitable amount Water for injection Amount required to make the total amount 1000 ml pH 5.5

Formulation Example 3 1 g of propylene glycol in 500 mL of water for injection,
0.05 g of benzalkonium chloride, 3.6 g of KH2
PO4 and 14.3 g of Na2HPO4.12H2O
Add and dissolve. Next, 50,000 units of UTI are added and mixed well. After adjusting the pH to 7.0 with 1N NaOH, water for injection is added to bring the total volume to 1000 mL.
The prepared propylene glycol-containing UTI aqueous solution was filtered and overfilled into a plastic eye drop bottle using a membrane filter having a pore size of 0.45 μm, and then subjected to intermittent sterilization to prepare a UTI-containing aqueous eye drop of the present invention. The prescription is shown below.

Ingredient UTI 50 units / mL Propylene glycol 0.1 w / v% KH2PO4 0.36 w / v% Na2HPO4.12H2O 1.43 w / v% Benzalkonium chloride 0.005 w / v% 1N NaOH Appropriate amount Water for injection Amount required to make 1000 ml pH 7.0

Formulation Example 4 5 g of propylene glycol and 5 g of chlorobutanol are added to 500 mL of water for injection, and dissolved while heating to 40 ° C. After cooling to room temperature, 1.2 g of acetic acid and 7.5 g of sodium acetate are added and dissolved. Further, add 15,000,000 units of UTI and mix well. After adjusting the pH to 6.0 with 1N HCl, water for injection is added to bring the total volume to 1000 mL. The prepared propylene glycol-containing UTI aqueous solution was filtered and filled into a dropping bottle using a membrane filter having a pore size of 0.22 μm to prepare a UTI-containing aqueous eardrop of the present invention. The prescription is shown below.

Component UTI 15000 units / mL Propylene glycol 0.5 w / v% Acetic acid 0.12 w / v% Sodium acetate 0.75 w / v% Chlorobutanol 0.5 w / v% 1N HCl Appropriate amount Water for injection Make up to 1000 ml Required for pH 6.0

Formulation Example 5 0.26 g of methylparaben and 0.14 g of propylparaben were added to 2 g of propylene glycol, and
Dissolve while heating to 0 ° C. Here is 500m of water for injection
L, followed by 5 g of ε-aminocaproic acid and 5.9 g
And dissolve while warming to 40 ° C.
After cooling to room temperature, 10,000,000 units of UTI
And mix well. PH 5.5 with 1N HCl
After adjusting to, water for injection is added to make the total volume 1000 mL. The prepared propylene glycol-containing UTI aqueous solution was filtered and filled into a nasal metering sprayer using a membrane filter having a pore size of 0.22 μm to prepare a UTI-containing aqueous nasal drop of the present invention. The prescription is shown below.

Component UTI 10000 units / mL Propylene glycol 0.2 w / v% Methyl paraben 0.026 w / v% Propyl paraben 0.014 w / v% ε-Aminocaproic acid 0.5 w / v% NaCl 0.59 w / v% 1N HCl Appropriate amount Water for injection Amount required to make the total amount 1000 ml pH 5.5

Formulation Example 6 The aqueous propylene glycol-containing UTI solution prepared in Formulation Example 5 was filtered and filled into an electric nebulizer using a membrane filter having a pore size of 0.22 μm to prepare a UTI-containing aqueous inhalant of the present invention.

Formulation Example 7 The propylene glycol-containing UTI aqueous solution prepared in Formulation Example 5 was filtered and filled into an atomizer using a membrane filter having a pore size of 0.22 μm to prepare a UTI-containing aqueous spray of the present invention.

Formulation Example 8 The aqueous propylene glycol-containing UTI solution prepared in Formulation Example 5 was filtered and filled into a dropping bottle using a membrane filter having a pore size of 0.22 μm to prepare a UTI-containing aqueous liniment or lotion of the present invention.

Formulation Example 9 10 g of propylene glycol in 500 mL of water for injection,
5 g of ε-aminocaproic acid and 5 g of sorbic acid are added and dissolved. In addition, 50,000 in UT
Add I and mix well. 4. Adjust pH to pH with 1N HCl.
After adjusting to 5, add water for injection to make the total volume 1000 mL. The prepared propylene glycol-containing UTI aqueous solution was filtered and overfilled into a glass bottle with a lid using a membrane filter having a pore size of 0.45 μm, and then subjected to intermittent sterilization treatment to prepare a UTI-containing aqueous oral liquid preparation of the present invention. The prescription is shown below.

Component UTI 50,000 units / mL Propylene glycol 1.0 w / v% Sorbic acid 0.5 w / v% ε-Aminocaproic acid 0.5 w / v% 1N HCl Appropriate amount Water for injection required to make the total volume 1000 ml Amount pH 5.5

[0041]

According to the present invention, compared to the conventional preparations,
It has become possible to provide a UTI-containing aqueous preparation having excellent heat stability of UTI activity. The UTI-containing aqueous preparation of the present invention is expected to be applied to the treatment of various diseases for which the administration of UTI is effective.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) A61P 43/00 111 A61P 43/00 111 C07K 14/81 C07K 14/81 C12N 9/99 C12N 9/99 ( 72) Inventor Masafumi Takeuchi 1-5-3 Nihonbashi Muromachi, Chuo-ku, Tokyo

Claims (4)

[Claims] 1. An urinastatin-containing aqueous preparation containing propylene glycol. 2. A propylene glycol content of 0.0
The urinastatin-containing aqueous preparation according to claim 1, which is 5 to 4 w / v%. 3. The method according to claim 1, wherein the pH is from 5.0 to 7.0.
3. The ulinastatin-containing aqueous preparation according to any one of 2. 4. The dosage form according to claim 1, wherein the dosage form is any one of an oral preparation, an injection, an eardrop, a nasal drop, an eye drop, a spray, an inhalant, a liniment, and a lotion. 2. The aqueous preparation containing ulinastatin according to claim 1.