JPH07165605A - Activated protein C vial - Google Patents

Abstract

(57) [Summary] [Purpose] A vial containing a daily dose of activated protein C. [Structure] Activated protein C is 2,000 to 20,0
Vial containing 00 units.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a pharmaceutical form for intravenous infusion of activated protein C having anticoagulant activity.

[0002]

BACKGROUND OF THE INVENTION Protein C is a vitamin K-dependent glycoprotein, which is synthesized in the liver and circulates in plasma as a zymogen at a concentration of about 4 μg / ml. Protein C is a serine protease activated by the thrombin-thrombomodulin complex on vascular endothelium,
That is, it is converted into activated protein C (hereinafter sometimes abbreviated as APC). APC hydrolyzes both proteins, Factor Va, which is a cofactor for factor Xa-induced prothrombin activation (thrombin generation), and Factor VIIIa, which is a factor IXa-induced cofactor for factor X activation. Has a coagulating effect.

APC also has a fibrinolytic activity because it binds to a tPA inhibitor and suppresses the tPA inhibitory activity of the inhibitor.

[0004]

The present inventors have found that a form of protein C which is activated against some thrombosis, namely A
The administration of PC was examined, and since the administration can be carried out by a simple operation, a diligent study was conducted to find a drug form in which the daily dose was contained in one vial.

[0005]

The present invention provides: 1. An activated protein C vial containing 2,000 to 20,000 units of activated protein C, and Activated protein C 2,000-20,000 units, human serum albumin 100-1,000 mg, sodium citrate 24-240 mg, glycine 20-200
is an activated protein C vial containing 28 mg and 28-280 mg sodium chloride.

Activated protein C itself used in the present invention is well known in the art, and protein C (hereinafter sometimes abbreviated as PC) derived from blood or prepared by genetic engineering technique is treated with thrombin or thrombin-thrombomodulin complex. Those activated in vitro or those obtained by direct expression as APC by a gene manipulation technique can be used.

[0007] One unit of APC is defined as an amount that doubles the activated thromboplastin time (APTT) of normal human plasma, and is specifically measured as follows.

The APC activity is measured by adding a diluted sample to normal human plasma and measuring the APTT (second). When the measured value is twice the value of the control (buffer solution), the dilution ratio is determined as the APC of the sample. The value of activity.

<Procedure> The specimen is diluted with Veronal buffer containing 1% HSA (for example, 400, 500, 800, 1).
000 times). At 37 ° C., 100 μl of each of the control (buffer solution) or each dilution of the sample was added with 100 μl of normal human plasma (eg, Cytolol) and 100 μl of APTT reagent (eg, actin) at 15-second intervals and mixed, and after 2 minutes , 100 μl of 0.025 M CaCl ₂ is added and the coagulation time is measured.

<Calculation of activity> From the APTT value (Y) at each dilution ratio (X) of the control and the sample, 10 ³ / X and Y were obtained.
The linear regression equation and the correlation coefficient are calculated.

[0011] ^{Y = A (10 3 / X} ) + B control APTT twice the value (s) as ^{_{Y 1, X 1 = 10 3}} {(Y 1 -B) / A} from the obtained of X ₁ The value is the APC activity of the sample (unit / m
l).

The activated protein C is added in an amount of 2,000-
As a vial used for containing 20,000 units, a vial which is usually used for intravenous injection or drip in a medical field can be used, and is described, for example, in the Japanese Pharmacopoeia.

APC alone may be present in such a vial, but usually various stabilizers are made to coexist in order to stabilize APC. As such a stabilizer, a protein and various salts or amino acids are typically used alone or as a mixture, and albumin and immunoglobulin as proteins, sodium chloride and citrate as salts,
It is selected from the group consisting of phosphates. Examples of amino acids include glycine, lysine, and alanine.

Among such combinations, the combination of human serum albumin, sodium citrate, glycine and sodium chloride is particularly preferable.

When the activated protein C vial of the present invention is used for intravenous injection or infusion, 4 to 40 c of physiological saline or purified water for injection is added to one vial of the present invention.
c is added, and if necessary, it is further added to 200 to 1000 cc.
It may be dissolved in the water for infusion and used for infusion. Since the vial of the present invention has a daily dose of one, it is not necessary to mix a plurality of vials or divide them into a plurality. Therefore, there is no mistake in the dose and there is no troublesome operation of mixing and dividing, so that it can be easily used in the medical field.

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

[0017]

Example 1 Method for Producing APC As an example of the method for producing APC used in the present invention, a method for activating PC purified from normal human plasma with thrombin to obtain APC will be shown. That is, normal human fresh frozen plasma was subjected to affinity chromatography using a monoclonal antibody that recognizes the Gla-domain to obtain protein C.
Get (PC). It is activated with thrombin, subjected to chromatographic purification using an anion exchanger, further dialyzed, filtered and lyophilized. As a specific example, the applicant of the present application filed on October 29, 1993 “Human activated protein C preparation and its production method” (Japanese Patent Application No. 5-292499).
See.

A stabilizer is added to the APC thus obtained to prepare an APC preparation having the following composition. For specific examples of the production method, refer to the applicant's application filed on October 29, 1993, “Stabilizing Method and Stabilizing Composition of Protein C or Activated Protein C” (Japanese Patent Application No. 5-292500).

[0019]

[Table 1]

This is packed in a vial so that the APC is 2,000 to 20,000 units.

[0021]

Example 2 Experiment for Determining Dosage Unit 48 leukemia patients with DIC were divided into 3 groups,
APC 50 units, 150 units and 300 units / kg
Daily administration was performed daily, and the degree of general improvement of DIC and the general safety level were examined. The best overall improvement was 300 units / kg-day, and there was no problem with the overall safety.

From now on, 150 to 300 units / kg · day can be adopted.

Considering the weight of a child to an adult to be 10 to 70 kg, the daily dose should be 1,500 to 21,000 units / vial, preferably 2,000 to 20,000 units / vial. It is preferable to prepare in series.

[0024]

[Example 3] A female patient 23 years old and 50 kg in weight who has an acute lymphocytic leukemia as a basic disease. I. Was diagnosed with generalized intravascular coagulation (DIC), and its DIC score was 4. 15,000 units of A for this patient
Using a vial containing PC, APC was intravenously infused at 300 units / kg / day for 6 days. The DIC score at the end of administration was 2 points, and the degree of improvement by the DIC score was moderate improvement. Further, the degree of improvement by the FM test and the like was markedly improved, and the degree of coagulation was markedly improved. From this, the comprehensive judgment is effective for DIC, without side effects,
This administration was found to be extremely useful.

[0025]

Example 4 A case is a 72-year-old man. Pr from _1st year of Heisei ₁
He was admitted to the hospital for the purpose of scrutiny and medical treatment, with an increase in aneurysm observed in March, ₁₉₉₂ , while he was being followed up with tein C (PC) deficiency, hypertension and abdominal aortic aneurysm. A spindle-shaped aneurysm with thrombus formation extending from just below the renal artery to the bifurcation of the common iliac artery was observed.
Coagulation was suspected to occur with DIC. PC activity 48
%, Antigen amount 35%, markedly decreased, and the second son of the patient also P
Both C activity and the amount of antigen were decreased, and the patient was diagnosed with type I PC deficiency. Heparin 10,000 U / day, ticlopidine hydrochloride 200 mg / day, while controlling DIC, an active PC preparation (C
TC-111) was continuously infused intravenously at 200 U / kg / 24 hr for 6 days, and the aneurysm resection could be safely performed without complications such as thrombosis. Improvement of DIC was also observed 10 days after the operation. Replenishment of active PC preparation was effective in the operation of abdominal aortic aneurysm with PC deficiency as a basic disease and severe DIC.

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Claims (2)

[Claims] 1. Activated protein C is 2,000 to 2
Activated protein C vial containing 10,000 units. 2. Activated protein C 2,000 to 20,
000 units, human serum albumin 250-1,000 m
g, sodium citrate 60 to 240 mg, glycine 5
Activated protein C vial containing 0-200 mg and 70-280 mg sodium chloride.