JP2008120798A - Heparin preparation - Google Patents

Abstract

An object of the present invention is to provide a heparin preparation excellent in stability during heating and storage and suppressed in coloring, and a method for stabilizing and suppressing coloration of the heparin preparation.
A heparin preparation comprising heparin or a salt thereof, trishydroxymethylaminomethane or an acid addition salt thereof, and gluconic acid or a salt thereof, a method for stabilizing a heparin preparation, and a method for suppressing coloration .
[Selection figure] None

Description

  The present invention relates to a heparin preparation, a method for stabilizing the same, and a method for suppressing coloration.

  Heparin is a polysaccharide N-sulfate, N-acetyl and O-sulfate substitute consisting of D-glucosamine, D-glucuronic acid and L-iduronic acid, and was first found in the liver of dogs. Many are also present in the skin, thymus and others. Most commercial products are derived from the intestine, and the molecular weight is about 7000 to 25000. Clinically, heparin's anticoagulant activity and liposerum activity are used.

  Sodium salts, calcium salts, and the like are known as heparin salts, and are used as pharmaceuticals. Sodium salt is a Japanese pharmacopoeia product. Heparin sodium injection solution is a glass vial injection containing 1000 units / ml of heparin sodium, which is colorless to pale yellow clear aqueous, pH is 5.5 to 8.0, and the osmotic pressure ratio to physiological saline is It has a property of about 1 grade. The prefilled syringe preparation for heparin lock is a preparation filled with 10 or 100 units / ml of heparin sodium in advance, and is colorless to pale yellow clear aqueous, pH is 5.5 to 8.0, and the osmotic pressure ratio is It has a property of about 1 grade. The dialysis-only heparin preparation is a glass vial or prefilled preparation containing 250 or 500 units / ml of heparin sodium, which is colorless to pale yellow clear aqueous, pH 5.5 to 7.5, and osmotic pressure ratio 0 It has properties such as 9 to 1.1. Heparin calcium injection solution is a glass vial or prefilled injection containing 1000 or 25000 units / ml of heparin calcium, which is colorless to pale yellow clear aqueous, pH 6.0 to 8.0, osmotic pressure ratio is It has a property of about 1 grade.

  When the pH of an aqueous heparin solution becomes lower than 6 due to absorption of carbon dioxide in the air, the sulfuric acid group of heparin is released by hydrolysis, which further lowers the pH and accelerates hydrolysis. When the sulfate group of heparin is released, its physiological activity also decreases. In particular, in a high-concentration heparin solution such as heparin calcium injection solution, or a so-called prefilled type formulation in which a heparin aqueous solution is prefilled in a gas permeable plastic container or syringe such as polypropylene or polyethylene, pH reduction and activity Decrease is a problem.

  In addition, conventional heparin aqueous solutions undergo a heat treatment such as high-pressure steam sterilization to reduce pH, decrease activity, or color. Therefore, the above preparations are intermittently sterilized or sterilized by filtration and / or used as preservatives / preservatives, such as benzyl alcohol, paraoxybenzoic acid. Some have added preservatives such as methyl and propyl paraoxybenzoate (for example, heparin sodium injection-Wf (manufactured by Nipropharma Co., Ltd., Mitsubishi Pharma Corporation) Sales), heparin sodium injection "Ajinomoto" (manufactured and sold by Ajinomoto Pharma Co., Ltd.)). Since benzyl alcohol is adsorbed on a butyl rubber stopper, it is necessary to use a rubber stopper coated with Teflon (registered trademark) or the like, and since it is decomposed by light, it is necessary to store the preparation under light shielding (non- Patent Document 1).

In recent years, according to guidelines for prevention of pulmonary thromboembolism / deep venous thrombosis (venous thromboembolism) (Non-Patent Document 2), a method of subcutaneously injecting 5000 units of heparin every 8 or 12 hours for the prevention of deep venous thrombosis Is recommended. In the case of subcutaneous administration, it is generally considered that a smaller dose volume is better in terms of pain and absorbability for the patient. Therefore, the heparin preparation used for the above-mentioned prophylaxis preferably has a high concentration and a small volume. . In addition, a unit-dose type prefilled syringe that can be used once is preferable for the purpose of saving the trouble of preparation for administration and preventing erroneous administration and contamination of drugs. However, it is unclear whether existing formulations satisfy these conditions and have sufficient stability.

Heparin or a salt thereof is mixed with citric acid, phosphoric acid, carbonic acid or trishydroxymethylaminomethane (hereinafter abbreviated as “Tris”, including acid addition salts unless otherwise specified) or a salt thereof. Thus, there is a disclosure that long-term storage stability can be achieved without adding the above-mentioned preservative / preservative, and as an example, sodium citrate, phosphate buffer, or carbonate A water-soluble preparation containing sodium hydride at a concentration of 1 mM is disclosed (Patent Document 1).
There is also a disclosure that the heparin solution can be stabilized by blending calcium lactate with heparin (Patent Document 2). In Example 1 of this document, there is a description that a heparin solution containing calcium gluconate instead of calcium lactate could not prevent a significant decrease in the concentration and pH of heparin in a conventional sterilization treatment.
In addition, as a pharmaceutical, for example, water-soluble, in which sodium citrate, sodium hydrogen phosphate or sodium hydrogen carbonate is mixed with sodium heparin at a low concentration (10 to 1000 units / ml) and the above preservative / preservative is not added. Preparations are commercially available (for example, heparin Na lock 100 syringe (manufactured by Nipro Pharma Co., Ltd., sold by Mitsubishi Pharma Corporation), heparin sodium injection N “Ajinomoto” (manufactured and sold by Ajinomoto Pharma Co., Ltd.), Pemilock (registered trademark) ) 100 units / mL syringe (manufactured by Taiyo Pharmaceutical Co., Ltd., Ajinomoto Pharma Co., Ltd.)).

  However, there is no disclosure in the publicly known literature or the like regarding the stabilization of the heparin preparation and the suppression of coloring by adding tris and gluconic acid or a salt thereof to the heparin preparation, and in particular, a high concentration (for example, 5000 units / ml or more). There is no disclosure regarding stabilization of the heparin preparation and suppression of coloring.

Pharmaceutical Additives Handbook, Maruzen Co., Ltd., 1988, P316 Prophylactic Guidelines for Pulmonary Thromboembolism / Deep Venous Thrombosis (Venous Thromboembolism) Digest Version, Medical Front International Limited, 2004, p7 JP 2002-104976 A Japanese National Patent Publication No. 4-507108

An object of the present invention is to provide a heparin preparation that is excellent in stability during heating and storage regardless of the type and concentration of heparin or a salt thereof, and regardless of the material and shape of the heparin preparation, or in which coloring is suppressed. Another object of the present invention is to provide a stabilization method and a coloring suppression method.
Another object of the present invention is to provide a high-concentration, small-volume, sufficiently stabilized and color-suppressed heparin preparation that can also be used for subcutaneous administration, particularly a single-use unit-dosage type prefilled syringe preparation. .

As a result of diligent research to solve the above problems, the inventors of the present invention incorporated Tris or an acid addition salt thereof and gluconic acid or a salt thereof, regardless of the type and concentration of heparin or a salt thereof. Further, the present inventors have found that a heparin preparation which is excellent in stability during heating and storage regardless of the material and shape of the container of the heparin preparation can be prepared or in which coloring is suppressed, and the present invention has been completed. That is, the present invention
(1) A heparin preparation characterized by containing at least heparin or a salt thereof, Tris, and gluconic acid or a salt thereof.

In particular,
(2) The heparin preparation according to (1), wherein the salt of heparin is heparin calcium,
(3) The heparin preparation according to (1) or (2) above, wherein the concentration of heparin or a salt thereof is 5000 to 50000 international units / ml,
(4) the heparin preparation according to any one of (1) to (3), wherein the pH is 6 to 8,
(5) The heparin preparation according to any one of (1) to (4), wherein the salt of gluconic acid is calcium gluconate.
(6) The heparin preparation according to any one of (1) to (5), further comprising calcium hydroxide,
(7) The heparin preparation according to any one of (1) to (6) above, which does not contain a preservative,
(8) The heparin preparation according to any one of (1) to (7) above, which is a pharmaceutical composition.

The present invention also provides:
(9) A method for stabilizing a heparin preparation, characterized in that heparin or a salt thereof, Tris, and gluconic acid or a salt thereof coexist.
(10) The stabilization method according to the above (9), wherein the stabilization is at least one of pH reduction suppression during heating and / or storage, heparin activity reduction suppression, or insoluble matter formation suppression,
(11) A method for inhibiting coloration of a heparin preparation, wherein heparin or a salt thereof, Tris, and gluconic acid or a salt thereof coexist.
The present invention also provides:
(12) A prefilled syringe filled with the heparin preparation according to any one of (1) to (8) above.
The present invention also provides:
(13) A heparin preparation characterized by containing at least heparin or a salt thereof and tris and / or calcium gluconate.

According to the present invention, there is provided a heparin preparation which is excellent in stability during heating and storage, or whose coloring is suppressed regardless of the type and concentration of heparin or a salt thereof, and regardless of the material and shape of the container of the heparin preparation. Can be provided. More specifically, at least one or more of pH reduction, heparin activity reduction, insoluble matter formation or coloring of the heparin preparation due to heating, storage, light, vibration, etc. can be suppressed. That is, heat sterilization, for example, high-pressure steam sterilization is possible, and it does not necessarily contain a preservative / preservative like conventional heparin preparations, and does not necessarily require light shielding, and is stable for long-term storage at room temperature, or A heparin preparation in which coloring is suppressed can be provided.
In addition, it is possible to provide a high-concentration small-volume sufficiently stabilized and color-suppressed heparin preparation that can also be used for subcutaneous administration, and a unit-dose type prefilled syringe preparation that can be used once.

The present invention is described in detail below.
A first aspect of the present invention is a heparin preparation characterized by containing at least heparin or a salt thereof, Tris, and gluconic acid or a salt thereof.
Known heparins can be used. Examples of heparin salts include sodium salts and calcium salts. Heparin sodium is obtained from, for example, the liver, lungs, and intestinal mucosa of healthy edible animals. It is a white to grayish brown powder or granule, has no odor, is slightly soluble in water, and is almost soluble in ethanol or diethyl ether. It may be anything that has properties such as no hygroscopicity. Heparin calcium is obtained from, for example, the intestinal mucosa of healthy pigs, and may be any white to ash-brown powder having properties such as no odor and easy solubility in water. Heparin is prepared according to a known method. For example, a method disclosed in the Japanese Pharmacopoeia can be used. Sodium salt is available in the Japanese Pharmacopoeia, and calcium salt is available in the UK Pharmacopoeia or, for example, manufactured by CarboMer Inc., Yantai Dongcheng Biochemicals Co. Ltd. is there.

Moreover, low molecular weight heparin is also exemplified as the heparin of the present invention. This is depolymerized heparin obtained by decomposing heparin derived from bovine or porcine intestinal mucosa with hydrogen peroxide, cupric sulfate, etc., for example, generic names parnaparin (average molecular weight 4500-6500), dalteparin (average molecular weight) About 5000), danaparoid (average molecular weight about 5500), reviparin (average molecular weight about 4000), enoxaparin (average molecular weight about 4500) and tinzaparin (average molecular weight about 5500-7500).
In the present invention, heparin units are expressed in international units (IU) unless otherwise specified. The relationship between IU and USP-U, which is an American heparin unit, is 1IU≈0.877 USP-U.
Examples of the heparin preparation of the present invention include pharmaceutically acceptable heparin-containing solutions, and heparin-containing solid preparations (powder preparations, freeze-dried preparations, etc.) used by dissolving at the time of use, and solutions are preferred.
The concentration of heparin or a salt thereof in the heparin preparation of the present invention or a solution in which the preparation is dissolved is about 10 to 50000 units / ml, preferably about 1000 to 50000 units / ml, more preferably 5000 to 50000 units / ml. Illustrated.

As the solvent, water is preferable, but another solvent that does not inhibit the heparin activity, for example, an organic solvent such as alcohol can be added.
The pH of the heparin preparation of the present invention or the solution in which the preparation is dissolved is preferably about 6 to 8. By providing a buffer capacity of pH 6 or higher, the stability of the heparin preparation of the present invention can be secured and improved. Specifically, the pH of the heparin preparation of the present invention or a solution in which the preparation is dissolved can be adjusted to 6 or more, a known buffer that has a buffering capacity at the pH and does not form a poorly water-soluble calcium salt. An appropriate amount of the agent can be blended.
Tris is preferably, for example, Tris hydrochloride, and when the concentration of heparin or a salt thereof is about 10 to 50000 units / ml, the addition concentration is about 25 to 200 mM, preferably about 40 to 100 mM, more preferably 50 to 100 mM. The degree is illustrated. As another preferred embodiment, when the concentration of heparin or a salt thereof is about 5000 to 50000 units / ml, tris hydrochloride is added at an addition concentration of about 10 to 200 mM, preferably about 20 to 100 mM, more preferably 20 to 60 mM. What includes a degree is illustrated. Gluconic acid or a salt thereof is preferably, for example, calcium gluconate. When the concentration of heparin or a salt thereof is about 10 to 50000 units / ml, the addition concentration is about 7 to 66 mM, preferably 22 to 56 mM, more preferably Is about 22 to 35 mM. As another preferred embodiment, when the concentration of heparin or a salt thereof is about 5000 to 50000 units / ml, calcium gluconate is added at an addition concentration of about 7 to 66 mM, preferably about 7 to 56 mM, more preferably about 7 to 35 mM. What is included is exemplified.
As a combination of Tris and gluconic acid or a salt thereof, for example, a combination of Tris hydrochloride and calcium gluconate is preferable, and when the concentration of heparin or a salt thereof, preferably heparin calcium is about 5000 to 50000 units / ml, The concentration of each added is about 10 to 200 mM of tris hydrochloride and about 7 to 66 mM of calcium gluconate, preferably about 20 to 100 mM of tris hydrochloride and about 7 to 56 mM of calcium gluconate, more preferably about 20 to 30 of tris hydrochloride. A combination of about 60 mM and about 7 to 35 mM calcium gluconate is exemplified.

The second aspect of the present invention is a method for stabilizing a heparin preparation, characterized in that heparin or a salt thereof, Tris, and gluconic acid or a salt thereof coexist. Stabilization of a heparin preparation is to suppress at least one or more of a decrease in pH, a decrease in heparin activity, or formation of an insoluble matter due to heating, storage, light, vibration, or the like.

A third aspect of the present invention is a method for inhibiting coloration of a heparin preparation, characterized by coexisting heparin or a salt thereof, Tris, and gluconic acid or a salt thereof. The suppression of coloration of the heparin solution may be colorless to light yellow and clear in the solution state, and it is preferable that the absorbance at 1 cm length (hereinafter abbreviated as OD400 nm) at a wavelength of 400 nm is 0.3 or less.
Preferred embodiments such as the type and concentration of heparin or a salt thereof, dosage form, solvent, pH, tris and gluconic acid or a salt thereof in the heparin preparation of the second and third embodiments of the present invention are as described above. It is the same as that of one aspect.

A fourth aspect of the present invention is a heparin preparation characterized by containing at least heparin or a salt thereof, Tris, gluconic acid or a salt thereof, and calcium hydroxide. When the concentration of heparin or a salt thereof is about 10 to 50000 units / ml, the concentration of calcium hydroxide added is about 2.8 to 49 mM, preferably about 2.8 to 16 mM.
Preferred embodiments such as the kind and concentration of heparin or a salt thereof, dosage form, solvent, pH, tris and gluconic acid or a salt thereof in the heparin preparation of the fourth embodiment of the present invention are the same as those in the first embodiment. It is.

A fifth aspect of the present invention is a prefilled syringe filled with a heparin preparation characterized by containing at least heparin or a salt thereof, Tris, and gluconic acid or a salt thereof.
Preferred embodiments such as the kind / concentration of heparin or its salt, dosage form, solvent, pH, tris and gluconic acid or its salt in the prefilled syringe of the fifth aspect of the present invention are the same as in the first aspect above. It is.

Tris and gluconic acid or a salt thereof or calcium hydroxide may be dissolved simultaneously with heparin or a salt thereof (heparin powder), or may be added either before or after dissolving the heparin powder. In the case where Tris and gluconic acid or a salt thereof or calcium hydroxide is added after dissolution of the heparin powder, at least a few hours after dissolution of the heparin is exemplified before sterilization by heating.
Before or after dissolution of the heparin powder, the pH of the solution is adjusted to 10 or more with a pH adjusting agent such as sodium hydroxide or calcium hydroxide, tris and gluconic acid or a salt thereof or calcium hydroxide is added, and the pH of hydrochloric acid or the like is added. It is also possible to adjust the pH to about 6 to 8 with a regulator. In addition, the preparation liquid can be bubbled with nitrogen during preparation.

In the heparin preparation and the method for stabilizing the same of the present invention, the stability of the heparin preparation can be secured and improved by blending the heparin preparation with tris and gluconic acid or a salt thereof or further calcium hydroxide.
Moreover, in the heparin formulation of the present invention and the coloring suppression method thereof, coloring of the heparin formulation can be suppressed by blending Tris and gluconic acid or a salt thereof or further calcium hydroxide.

The heparin preparation of the present invention can further contain at least one selected from an antioxidant, an isotonic agent, a pH adjuster, a stabilizer, an adsorption inhibitor, an antiseptic, and a preservative, thereby suppressing pain. One or more of an agent, a local anesthetic, and a tissue disorder inhibitor may be contained. The addition of an antioxidant is preferable because it enhances the coloration-suppressing effect of tris and gluconic acid or a salt thereof or calcium hydroxide with a heparin preparation. Examples of the antioxidant include reducing sulfites such as sulfites, bisulfites, disulfites, sodium salts of thiosulfates, and potassium salts. As an isotonizing agent, for example, salts such as sodium chloride and sugars such as glucose may be added. The osmotic pressure ratio may be a physiologically acceptable level, and is preferably about 0.9 to 1.1. By adding about 66 mM calcium gluconate, the osmotic pressure ratio can be adjusted to about 1 without adding sodium chloride, which is preferable for patients with sodium intake restriction such as patients with hypertension and renal insufficiency. As a pH regulator, for example, sodium hydroxide, calcium hydroxide, potassium hydroxide, hydrochloric acid and the like may be added.

The heparin preparation of the present invention thus prepared is heated (for example, high-pressure steam sterilization), long-term storage (for example, stored at room temperature for 3 years), light exposure (for example, non-shielded for 3 years), vibration (for example, several hours to Excellent stability during transport for several months), or excellent coloration resistance.
The heparin preparation of the present invention can be heat-sterilized, can provide a safe preparation with guaranteed sterility, and can be stored at room temperature for a long time without blending preservatives and preservatives. A specific method of heat sterilization includes high pressure steam sterilization. Stability or difficulty coloration during high-pressure steam sterilization is ensured by blending the above stabilizer or coloring inhibitor. Suitable conditions for high-pressure steam sterilization include a temperature of 105 to 130 ° C. and a time of about 1 to 60 minutes.

Preservatives and preservatives blended in conventional heparin preparations such as benzyl alcohol, methyl paraoxybenzoate, propyl paraoxybenzoate and the like can be used as a medicine with reduced or no addition.
In addition, since the heparin preparation of the present invention has long-term storage stability at room temperature and stability against light exposure or hardly coloring, heparin is kept stable even when stored at room temperature without being shielded from light, and is prominent. Since coloring is not recognized, management during storage is easy.

  The effect of the pharmaceutical product of the heparin preparation of the present invention includes treatment of generalized intravascular blood coagulation syndrome, prevention of blood coagulation when using hemodialysis / artificial cardiopulmonary and other extracorporeal circulation devices, and blood coagulation when inserting a vascular catheter Prevention, prevention of blood coagulation during blood transfusion and blood test, thromboembolism (venous thrombosis, myocardial infarction, pulmonary embolism, cerebral embolism, limb thromboembolism, thromboembolism during and after surgery, etc. ) Treatment and prevention, prevention of coagulation of perfused blood during extracorporeal blood circulation (artificial kidney and heart-lung machine, etc.), prevention of blood coagulation in the intravenous indwelling route, and the like.

  The method for using the pharmaceutical product of the heparin preparation of the present invention can be carried out in accordance with the usage / dose described in the package insert or guideline (non-patent document 2) of heparin sodium or heparin calcium preparation already on the market. It can be appropriately changed depending on the case, symptoms, age, sex, and the like. For example, at the time of use, it is dissolved or diluted to about 10 to 30 units / mL as it is or with glucose injection solution, physiological saline, Ringer's solution, etc., and then injected intravenously, subcutaneously or intramuscularly. For prevention of deep vein thrombosis, 5000 units are administered subcutaneously every 8 or 12 hours. In addition, for prevention of venous thrombosis secondary to surgery or myocardial infarction, 5000 units are administered subcutaneously every 12 hours for 7 to 10 days. In the case of subcutaneous administration and intramuscular administration, a single administration volume is 5 ml or less, preferably 0.1 to 1 ml, more preferably 0.2 ml.

The dosage form of the preparation includes an existing glass vial, a glass or plastic syringe, or an injection filled in a simple plastic injection kit such as Uniject (registered trademark) (Nippon Becton Dickinson Co., Ltd.) can do. So-called pre-filled pre-filled with the concentration and amount used in order to reduce the time and effort during preparation, and to prevent dilution errors during preparation, mistakes in drug mix, bacterial contamination and decreased activity due to divided use and storage. -It can be a unit dose type preparation. The capacity, material, and shape of the container can be appropriately selected from the viewpoint of the filling amount and ease of use of the heparin preparation. It is preferable to reduce the gas in the voids during filling or to substitute with nitrogen because the stability is further improved.
For treatment and prevention of thromboembolism, a prefilled syringe filled with 5000 units / 0.2 ml, 10,000 units / 0.4 ml or 15000 units / 0.6 ml of heparin sodium or heparin calcium is preferable. In particular, a prefilled syringe filled with 5000 units / 0.2 ml of heparin calcium, particularly a prefilled syringe for subcutaneous administration is preferable for the prevention of venous thrombosis secondary to surgery or myocardial infarction.

  EXAMPLES Examples and experimental examples for explaining the present invention in more detail are given below, but the present invention is not limited by these.

Examples Trishydroxymethylaminomethane (trade name Trometamol; manufactured by Wako Pure Chemical Industries, Ltd.) is dissolved in about 80 ml of water, then 2.5 million units of heparin calcium powder is gradually added and dissolved, and completely dissolved. Until stirred. In the case of adding calcium hydroxide (manufactured by Kokusan Chemical Co., Ltd.), heparin calcium was dissolved and then gradually added and stirred for a certain time. Next, when calcium gluconate monohydrate (manufactured by Wako Pure Chemical Industries, Ltd.) or to make isotonic, a predetermined amount of sodium chloride (manac Co., Ltd.) is added, and then the pH is 7. A 1M aqueous hydrochloric acid solution was added dropwise until about 5. Water was added to the pH-adjusted solution to make 100 ml, and heparin calcium preparations having the compositions shown in Table 1 were prepared. During preparation preparation, the preparation solution was bubbled with nitrogen.

Experimental example 1
Each of the heparin calcium preparations having the composition shown in Table 1 prepared in the Examples was filled in 0.3 ml of a polypropylene syringe having a capacity of 1 ml and then subjected to high-pressure steam sterilization at 125 ° C. for 10 minutes. These prefilled syringes were stored under the conditions shown in Table 2. The pH of the heparin calcium preparation before and after storage was measured. The results are shown in Table 2.

グルコン酸カルシウムを欠く対照に比し、トリスおよびグルコン酸カルシウムを添加した実施例１〜３の試料ではｐＨの低下が抑制された。
本発明製剤は、７０℃で４日間安定であり、上記結果から少なくとも室温で２年間は安定であると予測することができる。
また、固形の本発明製剤について、同様の滅菌条件、保存条件の後に溶液としたものについても上記と同様の結果を示した。

Compared to the control lacking calcium gluconate, the samples of Examples 1 to 3 to which Tris and calcium gluconate were added showed a decrease in pH.
The preparation of the present invention is stable at 70 ° C. for 4 days, and it can be predicted from the above results that it is stable at least at room temperature for 2 years.
Moreover, about the solid this invention formulation, the result similar to the above was shown also about what was used as the solution after the same sterilization conditions and storage conditions.

Experimental example 2
A prefilled syringe filled with the heparin calcium preparation shown in Table 1 was prepared according to Experimental Example 1. These samples were stored at 70 ° C. for 4 days. OD 400 nm was measured as an index of the degree of coloring of the heparin calcium preparation before and after storage. The difference in OD400 nm before and after storage of each sample (control and example) was calculated and used as the coloring degree by storage. The suppression rate of each sample with respect to the coloring degree by storage of the control was calculated based on the following formula. The results are shown in Table 3.

Inhibition rate (%) = (A−B) / A × 100
Where A: difference in OD 400 nm before and after storage of the control
B: Difference in OD400 nm before and after storage of each sample (Example)

表３に示すとおり、トリスおよびグルコン酸カルシウムを添加することにより、トリスのみを含む対照に比し、着色が顕著に抑制された。
また、同様にして、５０℃、６週間保存した各試料について、目視による確認を実施した。その結果、対照の試料には明らかな着色が認められたが、実施例１〜４の試料には着色は認められなかった。また、実施例１〜４の試料には、濁り、不溶物、沈殿物、析出物等は認められなかった。
また、固形の本発明製剤について、同様の滅菌条件、保存条件の後に溶液としたものについても上記と同様の結果を示した。

As shown in Table 3, coloring was remarkably suppressed by adding Tris and calcium gluconate as compared to a control containing only Tris.
Similarly, visual confirmation was performed for each sample stored at 50 ° C. for 6 weeks. As a result, clear coloring was observed in the control sample, but no coloring was observed in the samples of Examples 1 to 4. Further, turbidity, insoluble matters, precipitates, precipitates, and the like were not observed in the samples of Examples 1 to 4.
Moreover, about the solid this invention formulation, the result similar to the above was shown also about what was used as the solution after the same sterilization conditions and storage conditions.

Claims (12)

  A heparin preparation comprising heparin or a salt thereof, trishydroxymethylaminomethane or an acid addition salt thereof, and gluconic acid or a salt thereof.   The heparin preparation according to claim 1, wherein the salt of heparin is heparin calcium.   The heparin preparation according to claim 1 or 2, wherein the concentration of heparin or a salt thereof is 5000 to 50000 international units / ml.   The heparin preparation according to any one of claims 1 to 3, which has a pH of 6 to 8.   The heparin preparation according to any one of claims 1 to 4, wherein the salt of gluconic acid is calcium gluconate.   The heparin preparation according to any one of claims 1 to 5, further comprising calcium hydroxide.   The heparin preparation according to any one of claims 1 to 6, which does not contain a preservative.   The heparin preparation according to any one of claims 1 to 7, which is a pharmaceutical composition.   A method for stabilizing a heparin preparation, comprising combining heparin or a salt thereof, trishydroxymethylaminomethane or an acid addition salt thereof, and gluconic acid or a salt thereof.   The stabilization method according to claim 9, wherein the stabilization is at least one of pH reduction suppression during heating and / or storage, heparin activity reduction suppression, or insoluble matter formation suppression.   A method for inhibiting coloration of a heparin preparation, comprising coexisting heparin or a salt thereof, trishydroxymethylaminomethane or an acid addition salt thereof, and gluconic acid or a salt thereof.   A prefilled syringe filled with the heparin preparation according to any one of claims 1 to 8.