WO2018092241A1 - Method and apparatus for producing solution containing reduced albumin at increased content ratio - Google Patents

Abstract

Provided are: a method and an apparatus both for producing an albumin-containing solution in which the content ratio of reduced albumin is increased; and an albumin-containing solution having an increased reduced albumin content ratio, which is produced by the production method. A method for producing an albumin-containing solution, said method comprising (1) mixing an albumin-containing solution with a reducing agent having an SH group to produce a mixed solution and (2) dialyzing the mixed solution to produce an albumin-containing solution in which the content ratio of reduced albumin is increased, and said method optionally including (3) controlling the concentration of albumin. In the albumin-containing solution produced by the method, the content ratio of reduced albumin can be maintained at an increased level for a long period.

Description

Method and apparatus for producing solution with increased percentage of reduced albumin

The present invention relates to an easier and simpler method for producing a solution containing albumin containing an increased percentage of reduced albumin and a production apparatus, and a solution containing albumin containing an increased percentage of reduced albumin produced using the production method. About. The solution containing albumin having an increased ratio of reduced albumin produced by the production method and apparatus according to the present invention can maintain the increased ratio of reduced albumin for a longer period of time.

Albumin (alb) is a protein composed of about 600 amino acids and having a molecular weight of about 66,000. It accounts for about 60% of plasma proteins and has physiological functions such as maintaining plasma colloid osmotic pressure and transporting substances. .
Albumin is a reduced albumin (HMA) in which the SH group of the 34th cysteine residue from the N-terminus is free, and disulfide (SS) bonded to blood cysteine (Cys), glutathione, etc. Present as a mixture with oxidized albumin (HNA). According to the study of the present inventor, albumin in the blood of a healthy person contains about 70% reduced form and about 30% oxidized form. However, it has been reported that the ratio of reduced albumin decreases in cirrhosis patients, chronic renal failure patients, diabetic patients, patients under anesthesia and surgery, and the like.

Therefore, in recent years, not only the total amount of albumin but also the proportion of reduced albumin (HMA%) and the proportion of oxidized albumin (HNA%) in the whole albumin have attracted attention. In patients with various diseases, since HMA% decreases, it has been studied to increase HMA% (see Patent Documents 1 to 3).

Patent Document 1 proposes an oxidized albumin reducing agent containing at least one kind of branched chain amino acid selected from isoleucine, leucine, and valine as an agent that reduces HNA% in vivo (Patent Document 1 claims) Range). Specifically, granules with a weight ratio of isoleucine, leucine and valine of 1: 2: 1.2 were taken by 7 cirrhotic patients three times a day after meals, and the HNA% in their serum albumin was measured. It was done. The HNA% at the start of administration was about 40%, but it became about 33% after 8 weeks (see Patent Document 1 Example 1 and FIG. 1).

In Patent Document 2, when an albumin-containing intravenous agent prepared by mixing a first intravenous agent containing albumin as an active ingredient and a second intravenous agent containing cysteine or a similar compound thereof is used, the HMA% increases. Therefore, it is proposed that a higher therapeutic effect can be expected (see claims of Patent Document 2 and [0011]). Specifically, an intravenous albumin preparation having about 30% HMA% was mixed with strong Neominophagen C (registered trademark) intravenous injection containing cysteine hydrochloride, and the change in HMA% was examined. 10 minutes after mixing, the HMA% increased to about 64%, and after 15 minutes, the HMA% reached about 70% (maximum value). However, in 60 minutes, the HMA% was about 66%, and after 9 hours, the HMA% was reduced to about 40% (see Example 1 of Patent Document 2).
In Patent Document 2, more specifically, an albumin-containing intravenous agent obtained by mixing a first intravenous agent containing albumin as an active ingredient and a second intravenous agent containing cysteine or a similar compound thereof. Is directly administered to a patient by infusion (see Patent Document 2, claims 9 to 16, [0061] to [0072], see FIGS. 1 to 6).

Patent Document 3 discloses the prevention or treatment of edema associated with liver disease, which comprises the step (1) of providing albumin with an HNA% of less than 60% and the step (2) of dissolving the albumin of step (1) in an aqueous solvent. The manufacturing method of the albumin preparation for medical use is described (refer patent document 3 [0064]).
In the method of Patent Document 3, in the step (1), when cysteine is used as a reducing agent to produce albumin having an HNA% of less than 60%, a low molecule containing unreacted cysteine from the mixture after the reduction reaction Removing the compound and purifying albumin having a HNA% of less than 60%, and in step (2), the purified albumin is dissolved in an aqueous solvent, and 10 to 50 mg / 100 mL of cysteine is used as the reduced-type maintenance agent. Addition (see Patent Document 3 [0069]). Albumin is purified by chromatography or ultrafiltration, and a formic acid-acetonitrile aqueous solution is used as an extract (Patent Documents 3 [0057] and [0085] to [0087]).
Patent Document 3 shows that HNA% and HMA% can be quantified stably and with high accuracy using HPLC by purifying albumin (see Patent Document 3 [0021]).

Japanese Patent Laid-Open No. 2015-28052 WO2012 / 1210994A1 WO2009 / 123162A1

By the way, albumin preparations manufactured from human blood are often used to supplement albumin of patients who have been consumed due to liver disease or various invasions. Albumin preparations are classified into two types, “heated human plasma protein” and “human serum albumin”, and albumin purity and albumin concentration based on the total protein are defined. According to the study by the present inventors, the HMA% of the albumin (Alb) preparation is about 39% to about 42%, which is clearly lower than the above-described HMA% of blood albumin of healthy individuals. It is considered that reduced albumin was oxidized due to the physical background of the blood donor, the step of separating the albumin fraction from plasma, and natural oxidation during storage.
If the form (concentration and purity) of the albumin preparation is maintained as it is and the HMA% can be increased and the increased HMA% can be maintained for a long period of time, it is expected that the albumin preparation can be used as it is for the various patients described above. Is done.

On the other hand, patients with refractory ascites associated with end-stage liver cirrhosis and cancerous peritonitis have a feeling of bloating due to ascites retention and difficulty breathing. However, ascites drainage results in the loss of valuable proteins in the patient's body. That is, there is a problem that albumin is also lost.
Therefore, ascites filter (for example, AHF-MO (trade name) manufactured by Asahi Kasei Medical) and ascites concentrator (for example, AHF-UNH (trade name) manufactured by Asahi Kasei Medical) are not required for filtration from the patient's ascites. Cells, etc. are removed, excess water is removed (eg, concentrated to about 10 to about 15 times), and the ascites after filtration and concentration adjusted for albumin concentration, etc. is returned to the patient's blood. Cell-free and concentrated ascites reinfusion therapy (hereinafter also referred to as “CART”) has been performed.

The inventor measured the HMA% of albumin in the ascites of patients with CART. The HMA% of albumin in the ascites of patients with liver damage was significantly low (about 3.8% to about 20.1%). Albumin is synthesized in the liver. It is considered that albumin produced in the liver whose function has deteriorated due to liver damage is low, and it circulates in the patient's body for a long time, resulting in a decrease in HMA%.
Therefore, if the HMA% of albumin of ascites in a CART patient is increased and the increased HMA% can be maintained for a long period of time, it is expected that the ascites can be usefully used for the patient.

In Patent Document 1, it is unclear whether albumin having a low HMA% can be maintained for a long period of time by increasing the HMA% and increasing the HMA%. Moreover, the drug of Patent Document 1 must be taken after meals three times daily for a long period of time (specifically for two months).

In patent document 2, the intravenous injection which increased HMA% is obtained by adding a reducing agent to an albumin formulation.
However, the increased HMA% is not maintained for a long time. In our study, 3 days after adding the reducing agent, the HMA% returns to the original value (eg, about 30% to about 36%). The effect of the intravenous injection of Patent Document 2 cannot be expected to last for one day.

In Patent Document 3, albumin mixed with cysteine is purified by ultrafiltration or chromatography. Since formic acid-acetonitrile aqueous solution is used for purification, it is difficult to administer to humans. Acetyltryptophan and cysteine are considered to be added for the purpose of maintaining the stability and reduced state of albumin (see Patent Document 3 [0062]). However, since these compounds are not always necessary for patients, it is preferable not to include them (see Patent Documents 3 [0088] to [0089]). And even if acetyltryptophan and cysteine are added, it is unclear whether the increased HMA% is maintained for a long time, and the persistence of the effect is also unclear.

Furthermore, oxidized albumin (HNA) includes type 1 (HNA1) and type 2 (HNA2). Therefore, the ratio of HNA (HNA%) is the sum of the ratio of HNA1 (HNA1%) and the ratio of HNA2 (HNA2%) (HNA% = HNA1% + HNA2%). Reduced albumin (HMA) and type 1 oxidized albumin (HNA1) can be converted into each other. Type 1 oxidized albumin (HNA1) can be converted to type 2 oxidized albumin (HNA2), but HNA2 is not converted to HNA1. Therefore, it is considered important to prevent conversion from HNA1 to HNA2, that is, to suppress an increase in HNA2%. This is because once converted to HNA2, it cannot be converted to HNA1, and as a result, it cannot be converted to HMA. Therefore, there is a need for a method that does not convert HNA1 to HNA2 (or that is difficult to convert). Patent Documents 1 to 3 do not mention that HNA includes HNA1 and HNA2, and HNA1% and HNA2%.

As a result of intensive studies, the present inventor surprisingly increased the ratio of reduced albumin (HMA%) by adding a reducing agent having an SH group to the albumin solution, mixing, and dialysis. It has been found that an albumin solution can be obtained and that the increased HMA% can be maintained over a long period of time.

That is, the present invention in one aspect,
(1) Mixing an albumin-containing solution with a reducing agent having an SH group to produce a mixed solution; and (2) Dialyzing the mixed solution to contain albumin in which the ratio of reduced albumin is increased. Producing a solution, comprising:
(3) adjusting the concentration of albumin in the solution containing albumin in which the proportion of reduced albumin is increased,
Provided is a method for producing a solution containing albumin in which the ratio of reduced albumin is increased.

In another aspect, the present invention provides a solution containing albumin produced by the production method of the present invention and having an increased proportion of reduced albumin.

In a further aspect, the present invention provides:
(I) A mixing unit that mixes an albumin-containing solution and a reducing agent having an SH group to produce a mixed solution; and (II) dialyzes the mixed solution to obtain albumin having an increased ratio of reduced albumin. A dialysis part for producing a solution containing,
(III) A control unit for adjusting the albumin concentration of the solution containing albumin in which the ratio of reduced albumin is increased may be further included.
An apparatus for producing a solution containing albumin in which the ratio of reduced albumin is increased is provided.

The solution containing albumin with increased HMA% can be more easily and easily produced by the method and apparatus for producing a solution containing albumin with an increased ratio of reduced albumin (HMA%) according to the present invention. A solution containing albumin with increased HMA% can maintain the increased HMA% for a long period of time. The albumin-containing solution can be used for a patient as it is, preferably without requiring special treatment or addition of additives. A solution containing albumin with an increased HMA% can maintain the blood colloid osmotic pressure, which is the original role of albumin, and the ability to transport substances, for a long time, and may contribute to the improvement of the disease state.

Furthermore, when the production method according to the present invention is used, ascites (or ascites after filtration and concentration) with increased HMA% can be produced, and ascites can maintain the increased HMA% for a long time. Therefore, since the treated ascites (or ascites after filtration concentration) can be used for the patient himself by using the production method of the present invention for the CART case patient, albumin derived from the patient himself can be used. Therefore, it can be returned to the patient's own blood vessel as it is. Since the HMA% of albumin contained in the ascites increases and the increase can be maintained for a long time, the maintenance of blood colloid osmotic pressure, which is the original role of albumin, and the ability to transport substances can be maintained for a long time. There is a possibility of further contributing to the improvement of the disease state.

Moreover, using the manufacturing method according to the present invention, an albumin preparation capable of suppressing an increase in the ratio of type 2 oxidized albumin (HNA 2%) while increasing (or decreasing) HMA% is produced. Can do. The albumin preparation can maintain its HMA% for a long time and suppress the increase of 2% of HNA. Therefore, when an albumin preparation is produced using the production method of the present invention, the increased HMA% can be maintained, and the maintenance of colloid osmotic pressure, substance transportation and antioxidant action, which are the original roles of albumin, are maintained for a long time. It may also contribute more to the improvement of the patient's condition.

FIG. 1 shows an apparatus for producing a solution containing albumin in which the ratio of reduced albumin according to one embodiment of the present invention is increased. FIG. 2 shows an apparatus for producing a solution containing albumin in which the ratio of reduced albumin according to another embodiment of the present invention is increased.

Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings. However, the present invention is not limited by these forms.

A method for producing an albumin solution in which the ratio of reduced albumin (HMA%) in the form of the present invention is increased,
(1) Mixing an albumin-containing solution with a reducing agent having an SH group to produce a mixed solution; and (2) Dialyzing the mixed solution to contain albumin in which the ratio of reduced albumin is increased. Producing a solution,
(3) It may include adjusting the concentration of albumin.

In the step (1), an albumin-containing solution as a raw material and a reducing agent having an SH group that reduces oxidized albumin are mixed to produce a mixed solution.
The albumin-containing solution (raw material) is a solution containing at least albumin as an active ingredient, and is not particularly limited as long as a solution containing albumin with an increased HMA% targeted by the present invention can be obtained.
The solvent of the albumin-containing solution is preferably an aqueous medium, and examples of the “aqueous medium” include sterilized water, physiological saline, and physiological buffer.

The albumin-containing solution is pharmaceutically acceptable additive (for example, vitamins, electrolytes, trace elements, nutrients, stabilizers, oxidation agents, and the like) as long as the solution containing albumin with an increased HMA% targeted by the present invention can be obtained. An inhibitor, etc.).

The albumin-containing solution (raw material) is not particularly limited. For example, human albumin preparations such as “heated human plasma protein” and “human serum albumin”, ascites from patients with ascites, plasma, and genetic engineering techniques. A solution containing the produced recombinant albumin can be exemplified.
Examples of albumin preparations include, for example, Red Cross Albumin 25% intravenous injection (trade name) manufactured by the Japanese Red Cross, Venesis Blood Donation Albumin 25% intravenous injection (trade name), and Blood Donation Albumin 5% intravenous injection (trade name). Blood donation albumin 25 (trade name) manufactured by Kensha Co., Ltd. Blood donation albumin 25 (trade name) and blood donation albumin 5 (trade name) manufactured by Nippon Pharmaceutical Co., Ltd., Albuminer 25% intravenous injection (trade name) and albumin 5 manufactured by CSL Behring % IV injection (trade name), 25% Buminate intravenous injection (trade name) manufactured by Baxter, 5% buminate intravenous injection (trade name), and the like.
Ascites of ascites patients includes, for example, ascites of patients with ascites associated with end-stage cirrhosis, cancerous peritonitis and the like.
The plasma includes, for example, a fresh frozen plasma preparation and the like.
The solution containing recombinant albumin produced by genetic recombination technology includes Medway injection manufactured by Mitsubishi Tanabe Pharma.

The albumin concentration of the albumin-containing solution (raw material) is preferably 0.1 to 30 g / 100 mL, more preferably 0.5 to 25 g / 100 mL, from the fluidity of the albumin-containing solution and the efficiency of albumin utilization. A range of 5 to 25 g / 100 mL is particularly preferable.
The HMA% of the albumin-containing solution as a raw material is not particularly limited. The HMA% of the albumin-containing solution is preferably 60% or less, and more preferably 45% or less. In the case of ascites of ascites patients as a raw material, it is preferably 2 to 35%, particularly preferably 4 to 25%.

Examples of the “reducing agent having an SH group” include cysteine and its derivatives. The reducing agent having an SH group preferably contains cysteine. Examples of cysteine include L-cysteine, D-cysteine and DL-cysteine, and more preferably includes L-cysteine. Examples of cysteine derivatives include cysteine modified with a moiety other than the SH group, and include, for example, N-acetylcysteine and cysteine salts (such as cysteine hydrochloride).
The reducing agent having an SH group can be mixed with the albumin-containing solution in the form of a solution. The solvent is preferably the above-mentioned aqueous medium.

The reducing agent having an SH group can be used as it is as a solid, but can be used as a solution. As long as the solution can be obtained as an albumin solution with an increased HMA% as intended by the present invention, pharmaceutically acceptable additives (for example, vitamins, electrolytes, trace elements, nutrients, stabilizers, antioxidants, etc.) ) Can be included.
The concentration of the reducing agent having an SH group in the solution containing the reducing agent having an SH group is not particularly limited, but is preferably 0.05 to 5 g / 100 mL, preferably 0.08 to 3 g / 100 mL. More preferably, it is particularly preferably 0.2 to 1 g / 100 mL.

The molar ratio of the reducing agent having an SH group to albumin in the albumin-containing solution (reducing agent having an SH group / albumin) is preferably 0.2 to 5.0, and preferably 0.5 to 2.0. Is more preferable, and 1.0 to 2.0 is particularly preferable. When the molar ratio of the reducing agent having SH groups to the albumin of the raw albumin solution (reducing agent having SH groups / albumin) is 0.2 to 5.0, the conversion from HNA1 to HMA is more efficient. Since it is performed, it is preferable.

The mixing method of the reducing agent having an SH group and the albumin-containing solution is not particularly limited, and can be mixed using a commonly used method. As such a mixing method, for example, a method using a sterile syringe can be exemplified. The mixing conditions are not particularly limited, but for example, a temperature of 4 to 40 ° C, preferably a temperature of 4 to 35 ° C, more preferably 4 to 30 ° C, still more preferably 15 to 30 ° C, particularly preferably. May include a temperature of 20-30 ° C. A mixing time of 10 to 90 minutes is preferable, a mixing time of 15 to 90 minutes is more preferable, a mixing time of 20 to 75 minutes is more preferable, and a mixing time of 30 to 60 minutes is particularly preferable.

The manufacturing method of the embodiment of the present invention can have a filtration step (4). The filtration step (4) is preferably performed between the steps (1) and (2). In the filtration step (4), the liquid mixture of the albumin-containing solution and the reducing agent having an SH group can be filtered to remove unnecessary substances (such as cells) larger than albumin. Unnecessary substances can be discharged in the filtration process. In addition, filtration may be performed during the above-mentioned mixing time.

In the step (2), a mixed solution of an albumin-containing solution and a reducing agent having an SH group can be dialyzed to produce a solution containing albumin with increased HMA%.
It is preferable to dialyze (start dialysis) 10 to 90 minutes after mixing the albumin-containing solution and the reducing agent having an SH group, more preferably 15 to 90 minutes, and 20 to 75 minutes. It is more preferable to dialyze later, more preferably after 30 to 90 minutes, and particularly preferably after 30 to 60 minutes.

The dialysis method and conditions are not particularly limited as long as a solution containing albumin with an increased HMA% targeted by the present invention can be obtained. Depending on the albumin-containing solution, a dialysis method and conditions can be appropriately selected. For example, dialysis can be performed using a dialyzer and a dialysis device used for hemodialysis. A dialyzer using a hollow fiber membrane (hollow fiber type) is preferred, and a dialyzer using a polysulfone hollow fiber membrane is preferred. Furthermore, it is preferable to dialyze using an albumin non-leakage type dialyzer. The size of the pores of the hollow fibers contained in the albumin non-leakage type dialyzer is preferable because it is suitable for the size for blocking the albumin permeation.

The dialysis temperature is preferably 4 to 40 ° C., more preferably 4 to 35 ° C., still more preferably 4 to 30 ° C., still more preferably 15 to 30 ° C., and even more preferably 20 to It is particularly preferable that the temperature is 30 ° C.
The dialysis time is preferably 10 to 120 minutes, more preferably 20 to 90 minutes, and most preferably 30 to 60 minutes.
The flow rate of the mixed liquid is preferably 0 to 1000 ml / min, more preferably 40 to 700 ml / min, still more preferably 70 to 400 ml / min, and 100 to 200 ml / min. Is particularly preferred.
The dialysis fluid is preferably physiological saline, Ringer's solution, hemodialysis dialysate (or dialysate), and more preferably physiological saline or hemodialysis dialysate.
The flow rate of the dialysate is preferably 0 to 700 ml / min, more preferably 100 to 660 ml / min, still more preferably 250 to 630 ml / min, and 400 to 600 ml / min. Is particularly preferred.
Since dialysis can be performed by leaving the mixed solution and the dialysate separated by a semipermeable membrane, both the flow rate of the mixed solution and the flow rate of the dialysate can be 0 ml / min.

As the dialysis solution, a commercially available hemodialysis solution (or dialysis agent) can be used. The liquid dialysate can be appropriately diluted with water, and the powder dialysate can be appropriately dissolved in water for use. As such a dialysate, for example, a kinderial dialysate (trade name) manufactured by Fuso Pharmaceutical Co., Ltd., a kidryme dialysate (trade name) manufactured by Tomita Pharmaceutical Co., Ltd., a Rinpak dialysate (trade name) manufactured by Nipro Pharma Corporation, Nikkiso D dry dialysate (trade name) manufactured by Ajinomoto Co., Inc., Hisolv Dialyzer (trade name) manufactured by Ajinomoto Seiyaku Co., Ltd., and carboster dialysate (trade name) can be used.

As the dialyzer, a commercially available dialyzer for hemodialysis, a concentrator for ascites, and the like can be appropriately used depending on the albumin-containing solution as a raw material.
The membrane area of the dialyzer is preferably 0.0001 to 5.0 m ² , more preferably 0.001 to 3.0 m ² , still more preferably 0.01 to 2.0 m ² , Particularly preferred is 0.1 to 1.0 m ² .
Examples of such dialyzers include TS-S (trade name) manufactured by Toray Medical Co., Ltd., such as hollow fiber dialyzer (polysulfone dialyzer) APS-SA (trade name) and VPS-HA (trade name) manufactured by Asahi Kasei Medical. ), CS-S (trade name), etc., Fresenius FX (trade name), FX-S (trade name), etc., Nipro FB-EG (trade name), FB-UH (trade name), etc. Can be exemplified.

During the dialysis, the mixture of the albumin-containing solution and the reducing agent having an SH group passes through the dialyzer at least once. It is preferable to pass at least twice, more preferably at least 3 times, and particularly preferably 2 to 3 times.

In the step (3), the concentration of albumin can be further adjusted. The concentration can be lowered if the concentration of albumin is higher than the target concentration, and can be increased if the concentration of albumin is lower than the target concentration.
The albumin concentration can be adjusted using a known method. For example, a dialyzer, an ultrafiltration membrane, a vacuum freeze-drying apparatus, or the like can be used.
When the albumin-containing solution (raw material) is ascites, plasma or the like, it is preferable to increase the concentration of albumin.
A known method can be used as a method for measuring the concentration of albumin.

In this invention, the solution containing the albumin with which HMA% increased manufactured with the above-mentioned manufacturing method is provided.
The solution containing albumin with increased HMA% preferably contains 1-30 g / 100 mL of albumin, more preferably 2-30 g / 100 mL of albumin, and even more preferably 3-25 g / 100 mL. It is particularly preferable to contain 5 to 25 g / 100 mL.
The concentration of albumin in the solution containing albumin with increased HMA% may or may not correspond to the concentration of the albumin-containing solution (raw material), and is preferably an appropriate concentration. It can also be adjusted as appropriate.

The HMA% of the solution containing albumin with increased HMA% is preferably about 1 day at room temperature (temperature of 20-30 ° C.), preferably 2 days at 4 ° C. (total 3 days) from the end of dialysis. Later, it is preferably at least 10% higher than the HMA% of the albumin-containing solution (raw material), more preferably at least 15% higher, even more preferably at least 20% higher, even more preferably at least 25% higher, at least A height of 30% is particularly preferred.

The HMA% of the solution containing albumin with increased HMA% is preferably about 1 day at room temperature (temperature of 20-30 ° C.), preferably 2 days at 4 ° C. (total 3 days) from the end of dialysis. More preferably at least 50%, more preferably at least 55% after 6 days at 4 ° C. (total 7 days), or more preferably after 14 days (total 15 days). Even more preferably at least 60%, particularly preferably at least 65%, most preferably at least 70%.

The solution containing albumin with increased HMA% is preferably stored at room temperature (temperature of 20 to 30 ° C.) for 1 day from the end of dialysis, preferably further stored at 4 ° C. for 2 days (3 days in total). The ratio of reduced albumin is preferably 50% or more, more preferably 55% or more, still more preferably 60% or more, particularly preferably 65% or more, and 70% or more. Most preferably.

The solution containing albumin with increased HMA% is sealed at a temperature of 4 to 30 ° C., more preferably at a temperature of 20 to 30 ° C. for 1 day from the end of dialysis, and is kept in a state of being blocked from the outside air. Can be stored at normal pressure in the dark. Can be stored at 4 ° C (in the refrigerator), can be stored at -10 to -20 ° C (in the freezer), and stored at -60 to -80 ° C (in the deep freezer) be able to. Furthermore, it is preferable to preserve | save in the atmosphere of inert gas, such as nitrogen and argon, for example.

The solution containing albumin with increased HMA% is more preferably stored at a temperature of 4 ° C. (in a refrigerator) when stored for more than one day from the end of dialysis, and a temperature of −10 to −20 ° C. (in a freezer). It is more preferable to store at a temperature of −60 to −80 ° C. (in a deep freezer). It is preferable to store at normal pressure in a state of sealing and blocking from the outside air. Furthermore, it is preferable to preserve | save in the atmosphere of inert gas, such as nitrogen and argon, for example.

Since the solution containing albumin with increased HMA% is dialyzed, it can be used as it is in the patient without requiring special treatment or addition of additives.
By the way, for example, a human albumin preparation such as “human serum albumin” is used as an albumin-containing solution (raw material), the solution is mixed with a reducing agent having an SH group, and a mixed liquid in which MHA% is increased is obtained. Obtainable. Since a large amount of N-acetyltryptophan is added as an additive to the raw material albumin preparation, a large amount of the additive is also contained in the obtained mixed solution. It is preferable for the patient not to take such additives. In the present invention, the mixed solution is dialyzed. Since the additive N-acetyltryptophan is a small molecule, it can be removed from the mixture, and the resulting solution containing albumin with increased HMA% does not contain a large amount of N-acetyltryptophan, and is more preferable.
Furthermore, the solution containing albumin with increased HMA% may have a more physiologically favorable electrolyte concentration because it has been dialyzed.
The solution containing albumin with increased HMA% in the form of the present invention requires pharmaceutically acceptable additives (for example, vitamins, electrolytes, trace elements, nutrients, stabilizers, antioxidants, etc.). Can be included.

By the way, it is preferable not to add a reducing agent having an SH group to a solution containing albumin with increased HMA%. If unreacted HNA1 is present, further HMA is obtained by a reducing agent having HNA1 and an SH group (for example, cysteine). However, a reducing agent having an SH group (for example, cysteine) remains, from which a compound having a disulfide bond (for example, cystine) is produced, which can remain. HNA1 reacts with the compound having the disulfide bond to generate HNA1 again. Therefore, when storing for a long period of time, it is preferable not to add a reducing agent having an SH group (for example, cysteine or the like) to a solution containing albumin with increased HMA%.

The present invention further provides:
(I) A mixing unit that mixes an albumin-containing solution and a reducing agent having an SH group to produce a mixed solution; and (II) dialyzes the mixed solution to obtain albumin having an increased ratio of reduced albumin. Including a dialysis section to produce a solution containing,
(III) It may further include a regulator that regulates the albumin concentration,
An apparatus for producing a solution containing albumin in which the ratio of reduced albumin (HMA%) is increased is provided.

(I) “Albumin-containing solution”, “reducing agent having SH group”, “mixed solution”, “mixing” in the mixing part, (II) “dialysis”, “reduced albumin ratio in the dialysis part increased The “solution containing albumin” and the like are as described above.

(I) The mixing part is not particularly limited as long as a solution containing albumin with an increased HMA% targeted by the present invention can be obtained, and may be a commonly used mixing apparatus. As such a mixing part, a container, a bag, etc. which have a sterilization state can be illustrated, for example.
(I) The albumin-containing solution and the reducing agent having an SH group are mixed in the mixing unit. The mixing conditions are preferably the above-described molar ratio of the reducing agent having an SH group to albumin (reducing agent having an SH group / albumin), the mixing temperature, the mixing time, and the like.

The manufacturing apparatus according to the embodiment of the present invention can have a filtration unit (IV). The filtration part (IV) is preferably provided between the (I) mixing part and the (II) dialysis part. A filtration part has a filter and can filter the liquid mixture of an albumin containing solution and the reducing agent which has SH group, and can remove an unnecessary substance (for example, cell etc.) larger than albumin. Unnecessary substances can be appropriately discharged from the filter.

(II) The dialysis part is not particularly limited as long as it can obtain a solution containing albumin with an increased HMA%, which is the object of the present invention, and may be a commonly used dialysis machine. As such a dialysis part, a hemodialysis apparatus, an apheresis apparatus, etc. can be illustrated, for example. The dialysis unit can include a dialyzer, a dialysate supply device, and the like.

(II) The dialysis part dialyzes the mixed solution of the albumin-containing solution and the reducing agent having an SH group, and the dialysis conditions at that time are preferably the above dialysis temperature, dialysis time, dialysis solution, and the like.

(III) In the adjustment unit, the concentration of albumin can be further adjusted. The concentration can be lowered if the concentration of albumin is higher than the target concentration, and can be increased if the concentration of albumin is lower than the target concentration.
As the adjustment unit, a known concentration adjustment device can be used. Examples of such devices include a dialyzer, a vacuum freeze-drying device, an ultrafiltration membrane, and an apheresis device.
When the albumin-containing solution (raw material) is ascites, plasma or the like, it is preferable to increase the concentration of albumin by (III) the control unit.

FIG. 1 shows an apparatus 1 for producing a solution containing albumin in which the ratio (HMA%) of reduced albumin according to one embodiment of the present invention is increased.
The manufacturing apparatus 1 includes a mixing unit 10 and a dialysis unit 30. The mixing unit 10 includes a mixed solution 12 obtained by mixing a raw material albumin-containing solution and a reducing agent having an SH group. The dialysis unit 30 includes a dialysis dialyzer 32 and a dialysate supply device 34.
In the mixing part 10, the raw material albumin containing solution and the reducing agent having SH groups can be mixed to produce the mixed liquid 12. Furthermore, the liquid mixture 12 may be manufactured in advance in another place, and then the liquid mixture 12 may be transferred to the mixing unit 10. Alternatively, half of the mixed liquid 12 may be manufactured in another place, and the mixed liquid 12 may be manufactured as a whole by moving to the mixing unit 10 and subsequently manufacturing the rest. The mixed liquid 12 can be produced in the mixing unit 10 as appropriate. The mixing conditions in the mixing unit 10 are as described above.

From mixing of the raw material albumin-containing solution and the reducing agent having an SH group, preferably, after a predetermined time has elapsed, the mixed solution 12 is dialyzed through a dialysis dialyzer 32 of the dialysis unit 30. At that time, the dialysate 36 is supplied from the dialysate supply device 34 to the dialyzer 32 for dialysis. The mixed solution 14 that has passed through the dialyzer 32 returns to the mixing unit 10. The dialysate 38 that has passed through the dialyzer 32 returns to the dialysate supply device 34. The mixture 12 is further dialyzed repeatedly through the dialyzer.

In addition, when the flow rate of the dialysate 36 and the flow rate of the dialysate 38 are different, when the mixed solution 12 passes through the dialyzer 32 and the mixed solution 14 can be obtained, the albumin concentration of the mixed solution can be concentrated or diluted. it can. This can also be performed by making the flow rate of the mixed solution 12 different from the flow rate of the mixed solution 14. For example, if the flow rate of the dialysate 38 is made faster than the flow rate of the dialysate 36, a negative pressure is generated, so that the mixed solution is concentrated. On the contrary, it is diluted. Further, when the flow rate of the mixed solution 12 is made faster than the flow rate of the mixed solution 14, pressurization occurs, so that the mixed solution is also concentrated in this case. On the contrary, it is diluted. By adjusting the flow rates of the dialysates 36 and 38 and the flow rates of the mixed solutions 12 and 14, the albumin concentration of the mixed solution 14 can be adjusted as appropriate.
The production apparatus 1 of FIG. 1 can be used for a variety of raw material albumin-containing solutions. For example, it is preferably used for human albumin preparations such as “heated human plasma protein” and “human serum albumin”. Can do.

FIG. 2 shows an apparatus 2 for producing a solution containing albumin in which the ratio of reduced albumin (HMA%) according to another embodiment of the present invention is increased.
The manufacturing apparatus 2 includes a dialysis unit 70 that also serves as a mixing unit 50, a filtration unit 60, and a control unit.
The mixing unit 50 includes a mixed solution 56 obtained by mixing a raw material albumin-containing solution and a reducing agent having an SH group. In the mixing unit 50, the raw material albumin-containing solution 52 and the reducing agent 54 having an SH group can be mixed to produce a mixed solution 56. Further, the mixed liquid 56 may be manufactured in advance in another place, and then the mixed liquid 56 may be transferred to the mixing unit 50. These are the same as in FIG.
The filtration unit 60 includes a filter 62. The mixed liquid 56 is filtered by a filter 62 to give a mixed liquid 66 from which unnecessary substances larger than albumin (such as cells) are removed. Waste 68 containing insoluble matter is appropriately discharged from the filter 62.

The dialysis unit 70 also serving as the adjustment unit includes a dialyzer 72 and an adjustment / dialysis device 74. The liquid mixture 66 from the filtration unit 60 enters the dialyzer 72. A dialysate 75 is sent from the adjustment and dialysis device 74 to the dialyzer 72. Mixture 66 is dialyzed with dialyzer 72 and the concentration is adjusted to provide mixture 78. The dialysate 75 gives the used dialysate 76 from the dialyzer 72.
When the albumin concentration of the mixed solution 78 is high or low, the concentration of albumin or the like can be adjusted by repeatedly passing the dialyzer 72 as necessary.
It is preferable to adjust the concentration of the mixed solution 78 by repeatedly passing through the dialyzer 72 after the mixed solution 56 is filtered, concentrated and dialyzed to obtain the mixed solution 78 once.
The production apparatus of FIG. 2 can be preferably used when the raw albumin-containing solution 52 has a large unnecessary substance to be removed by filtration, and the albumin concentration is different from the desired concentration. In the manufacturing apparatus of FIG. 2, for example, the albumin-containing solution 52 is preferably ascites, a blood product, or the like.
The elements (lines, containers, bags, etc.) for handling the mixed liquids 12 and 14, dialysis liquids 36 and 38 in FIG. 1, the mixed liquid 56, mixed liquid 66, dialysis liquid 75, and mixed liquid 78 in FIG. It is more preferable to be closed in order to keep it from being sterilized by being shielded from outside air.

Further, when the production method according to the present invention is used in producing a human albumin preparation, the proportion of reduced albumin (HMA%) is increased (or not decreased) while the proportion of type 2 oxidized albumin is increased. A human albumin preparation can be produced while suppressing an increase in (HNA 2%).
According to the study of the present inventor, the HNA 2% of the blood of a healthy person is about 2% to about 3%. On the other hand, HNA 2% of the commercially available albumin preparation is about 15% to about 18%, which is much higher.
Usually, the albumin fraction is produced by extracting Cone from human plasma with ethanol and extracting from the final solution obtained by extracting components such as coagulation factors and fibrinogen. Therefore, during the manufacture of albumin preparations, it is considered that HNA 2% is increased due to various oxidative modifications.
Therefore, the production of the present invention can be used to maintain high HMA% at various stages such as collecting human plasma, extracting Cone with ethanol, and extracting components such as coagulation factors and fibrinogen. As a result, an increase in HNA of 2% can be suppressed.

For example, using the production apparatus 1 of FIG. 1, a method of using a solution obtained by collecting human plasma as a raw material albumin-containing solution 52, a method of using Cone ethanol extraction as a raw material albumin-containing solution 52, and coagulation. A method in which the final liquid obtained by extracting components such as factors and fibrinogen is used as the raw material albumin-containing solution 52 can be considered.
In any method, the obtained albumin solution 14 with increased HMA% can maintain high HMA%, and as a result, can suppress an increase in HNA2%.
Therefore, the albumin preparation obtained thereafter can also maintain a high HMA% and suppress an increase in HNA2%.
Therefore, when an albumin preparation is produced using the production method of the present invention, HMA% can be maintained high, and the maintenance of colloid osmotic pressure, substance transport and antioxidant action, which are the original roles of albumin, can be sustained for a long time. There is a possibility of further contributing to the improvement of the patient's condition.
Furthermore, since HNA 2% can be kept low, albumin can be used more effectively as a result.

The present invention has the excellent effects as described above, and the reason thereof is as follows. However, the present invention is not limited to the reason.
Concentrated ascites and albumin preparations of refractory ascites patients can increase HMA% by the method according to the present invention, and can maintain the increased HMA% over a long period of time.
From the results of the inventor's investigation (Comparative Example of the present application) and Patent Document 2, the increased HMA% cannot be maintained simply by adding a reducing agent, an antioxidant, or the like.
Healthy individuals have about 65% HMA% for their serum albumin and can maintain that value. This healthy person's HMA% value is significantly higher than the HMA% (about 6% to about 20%) of albumin in the ascites of refractory ascites patients and the HMA% (about 39% to about 42%) of albumin preparations.
Thus, even if the HMA% of albumin is increased, the present inventors show that when the environment surrounding albumin is an environment that stresses albumin or a pathological environment, HMA% I thought it would have a negative effect. On the other hand, if the environment surrounding albumin having increased HMA% can be replaced with the environment in the body of a healthy person, it was considered that the increased HMA% can be maintained over a long period of time.
As a result of various examinations for realizing the environment of the healthy person and dialysis, it was surprisingly found that HMA% can be maintained over a long period of time.

Hereinafter, the present invention will be described specifically and in detail with reference to examples and comparative examples, but these examples are only one aspect of the present invention, and the present invention is not limited to these examples.

(A) Albumin-containing solution and (B) a reducing agent having an SH group used in Examples and Comparative Examples are shown below.

(A) Albumin-containing solution (a1) 5% albumin preparation (Albminer 5% IV (trade name) manufactured by CSL Behring Co., Ltd., albumin concentration: about 0.75 mmol / L, according to measurement by the present inventor, HMA% : About 39% (see Comparative Example 8))
(A2) Ascites concentrate of patients with refractory ascites (ascites concentrate of 6 patients with low HMA%, albumin concentration: 15 μmol / L and about 1 g / L, according to measurement by the present inventor, HMA%: about 13. 5% (see Comparative Example 9))

(B) A reducing agent having an SH group (b1) A mixed solution of ascorbic acid and L-cysteine (Christophane Note (trade name) manufactured by Nissin Pharmaceutical Co., Ltd.), 20 mg of ascorbic acid in 1000 mg (284 mmol / L) and L-cysteine 80mg (33mmol / L)
(B2) L-cysteine solution (8.2 mmol / L aqueous solution of L-cysteine (trade name) manufactured by Wako Pure Chemical Industries, Ltd.)
(B3 ′) Ascorbic acid solution (340 mmol / L aqueous solution of ascorbic acid (trade name) manufactured by Wako Pure Chemical Industries, Ltd.)
Saline was used as the dialysate.

Example 1
(A1) 5% albumin preparation containing 12.5 g of albumin (250 mL, albumin: 0.19 mmol) and (b1) a mixture of ascorbic acid and L-cysteine (12 mL, ascorbic acid: 3.4 mmol, L-cysteine: 0.4 mmol). Albumin: L-cysteine = 1: 2.1 (molar ratio) and albumin: ascorbic acid = 1: 17 (molar ratio). After gently stirring at 25 ° C., the mixture was allowed to stand for 60 minutes to obtain a mixed solution. The mixture was dialyzed under the following conditions to obtain an albumin solution of Example 1 in which the ratio of reduced albumin was increased.
Using a hollow fiber type dialyzer (polysulfone hollow fiber dialyzer) (membrane area: 0.01 m ² ), the mixed solution is circulated at a flow rate of 50 ml / min, and 3 L of physiological saline is circulated at a flow rate of 125 ml / min. And dialyzed for 60 minutes. The appearance of a reducing agent having an unreacted SH group in the dialysate waste liquid was caused by mixing the DTNB reagent (200 μL) with the dialysate waste liquid (100 μL), and the color tone of the mixture of the reagent and waste liquid turned yellow. Confirmed by doing.

The ratio of reduced form of albumin (HMA%) in the mixture and dialysate was determined by a method using high performance liquid chromatography (HPLC) (Sogami M., J. Chromatogr., 332, 19-27, 1985; Int. J Peptide Protein Res. 24:96; 1984). That is, reduced albumin and oxidized albumin were separated and detected by HPLC, and HMA% was determined from the ratio of peak areas on the separated chromatogram.
More specifically, SCL-10A (trade name) manufactured by Shimadzu Corporation was used as the HPLC apparatus. As a column, ES-520N (trade name) manufactured by Showa Denko KK was used at a column temperature of 35 ° C. 0.05M sodium acetate, 0.4M sodium sulfate (pH 4.85) buffer and 100% ethanol were used. A 2 μL sample was analyzed. Initially, the buffer solution was flowed at a flow rate of 1.0 mL / min, and the flow rate of the buffer solution was reduced to 0.9 mL / min over 49 minutes. In contrast, initially, the flow rate of 100% ethanol at a flow rate of 0 mL / min was increased to 0.1 mL / min over 49 minutes. The total flow rate of the buffer and 100% ethanol was 1.0 mL / min in total. Detection was performed using a fluorescence detector having an excitation wavelength of 280 nm and a fluorescence emission wavelength of 340 nm. The results are shown in Table 1.
The albumin solution of Example 1 produced by adding a reducing agent having an SH group to albumin and then dialysis was stored in a refrigerator at 4 ° C. for 1 day at room temperature (20 to 30 ° C.) after dialysis. The increased HMA% could be maintained for a further 2 days and thus for a total period of 3 days or more.

Examples 2 to 4 and Comparative Example 1
(B1) In the same manner as in Example 1 except that the amount of the mixed solution of ascorbic acid and L-cysteine was decreased, (a1) a 5% albumin preparation and (b1) ascorbic acid and L-cysteine The mixed solution was mixed, and the obtained mixed solution was dialyzed to obtain albumin solutions of Examples 2 to 4 and Comparative Example 1.
That is, (a1) 5% albumin preparation (1250 mL) and (b1) a mixture of ascorbic acid and L-cysteine (30 mL, 15 mL, 7.5 mL, or 3.8 mL) were mixed.
Albumin: L-cysteine = 1: 1 (molar ratio) (Example 2), 1: 0.5 (molar ratio) (Example 3), 1: 0.25 (molar ratio) (Example 4) or 1 : 0.13 (molar ratio) (Comparative Example 1), albumin: ascorbic acid = 1: 8.5 (molar ratio) (Example 2), 1: 4.25 (molar ratio) (Example 3) 1: 2.13 (molar ratio) (Example 4) or 1: 1.06 (molar ratio) (Comparative Example 1).
The results are shown in Table 1.
The albumin solutions of Examples 2 to 4 manufactured by adding dialysis with more than 0.13 mol of L-cysteine per 1 mol of albumin had increased HMA%, and after completion of dialysis, room temperature (20 to 30 It was found that increased HMA% could be maintained for 1 day at 4 ° C. and then for an additional 2 days in a refrigerator at 4 ° C., thus over a total period of 3 days or more.

Comparative Examples 2-5
In the albumin solutions of Comparative Examples 2 to 5 obtained by the same method as in Examples 1 to 4 except that dialysis was not performed, the HMA% increased immediately after the albumin preparation and the cysteine solution were added. However, after mixing the albumin preparation and the cysteine solution, after 1 day at room temperature (20 to 30 ° C.), the HMA% rapidly decreases, and then in the refrigerator at 4 ° C. for another 2 days, thus a total of 3 It can be seen that after a day, HMA% returns to its original value.

*) (A)の数値は、アルブミンのｍｍｏｌを示す。
**) (B)の(b1)及び(b2)の数値は、システインのｍｍｏｌを示す。

*) The numerical value of (A) indicates mmol of albumin.
**) The numerical values of (b1) and (b2) in (B) indicate mmol of cysteine.

Example 5 and Comparative Examples 6-8
As described in Table 2, HMA, HNA1 and HNA2 were used in the same manner as described in Example 1 except that (b2) or (b3 ′) was used instead of (b1). Based on the following formula, HMA%, HNA1% and HNA2% were obtained from the area of each obtained peak.
HMA% = HMA / (HMA + HNA1 + HNA2) × 100
HNA1% = HNA1 / (HMA + HNA1 + HNA2) × 100
HNA2% = HNA2 / (HMA + HNA1 + HNA2) × 100
Then, HMA% of the mixed solution of Example 5 and Comparative Examples 6 to 8 was obtained. The results are shown in Table 2.
Only by adding a reducing agent having an SH group (L-cysteine) (Comparative Example 6), just adding another antioxidant (ascorbic acid) (Comparative Example 7), or just dialyzing (Comparative Example 8) Over time, high HMA% of albumin could not be maintained, and it was found that albumin was dialyzed after mixing with a reducing agent having an SH group (Example 5) to maintain high HMA% over time. .

When HNA 1% of Example 5 and HNA 1% of Comparative Example 6 are compared, it can be seen that HNA 1% of Comparative Example 6 increases with time, and the difference between the HNA 1% of Example 5 increases.
Comparing the HNA 2% of Example 5 with the HNA 2% of Comparative Example 6, it can also be seen that the HNA 2% of Comparative Example 6 increases with time, and the difference between the HNA 2% of Example 5 increases.
Therefore, it can be seen that dialysis not only suppresses an increase in HNA of 1% but also suppresses an increase in HNA of 2%. It can be considered that suppressing the increase of 2% of HNA contributes to maintaining a high HMA% in the long term.

*) (A)の数値は、アルブミンのｍｍｏｌを示す。
**) (B)の(b1)及び(b2)の数値は、システインのｍｍｏｌを示す。

*) The numerical value of (A) indicates mmol of albumin.
**) The numerical values of (b1) and (b2) in (B) indicate mmol of cysteine.

Example 6
(A2) An ascites concentrate (10 mL) of an intractable ascites patient and (b2) L-cysteine solution 8.2 mmol / L (2.6 mL) were mixed. Albumin: L-cysteine = 1: 2 (molar ratio) (in ascites concentrate). After gently stirring at 25 ° C. and allowing to stand for 60 minutes, the mixed solution was dialyzed under the following conditions to produce the albumin solution of Example 6 in which the HMA% of Example 6 increased.
Using a hollow fiber type dialyzer (polysulfone hollow fiber dialyzer) (membrane area: 0.001 m ² ), the mixture was circulated at a flow rate of 5 ml / min, and the dialysis solution was circulated at a flow rate of 15 ml / min. Dialyzed for 30 minutes. The appearance of a reducing agent having an SH group in the dialysate waste liquid by dialysis was confirmed by mixing the DTNB reagent (200 μL) and the dialysate waste liquid (100 μL) and changing the color tone to yellow.

The HMA% of the mixed solution and dialysate was determined using the method described in Example 1.
The results are shown in Table 2.
By adding a reducing agent having an SH group to ascites and then dialyzing, high HMA% can be maintained for albumin in ascites at a temperature of 20 to 30 ° C. for a long period of one day or longer after dialysis. I understood it.

Comparative Example 9
As described in Table 2, an albumin solution of Comparative Example 5 was produced using the same method as in Example 6 except that dialysis was not performed. The results are shown in Table 2.
It was found that if the ascites concentrate was not dialyzed after mixing with a reducing agent having an SH group, the HMA% of albumin decreased in a short time at a temperature of 20 to 30 ° C. after completion of dialysis.
Therefore, it was revealed that ascites containing albumin is important to be dialyzed after mixing with a reducing agent having an SH group.

The present invention provides a method and an apparatus for producing a solution containing albumin in which the proportion of reduced albumin is increased, and a solution containing albumin in which the proportion of reduced albumin produced using the production method is increased. A solution containing albumin having an increased ratio of reduced albumin produced by the method and apparatus according to the present invention can maintain the increased ratio of reduced albumin for a long period of time.

1: Solution manufacturing apparatus containing albumin with increased HMA% in one aspect of the present invention 2: Solution manufacturing apparatus including albumin with increased HMA% in another aspect of the present invention 10: Mixing unit 12: Albumin-containing solution Mixture 14 with a reducing agent having an SH group 14: Dialyzed mixture 30: Dialysis section 32: Dialyzing dialyzer 34: Dialysate supply device 36: Dialysate 38: Dialysate 50 passed through dialyzing dialyzer 50: Mixing Part 52: Albumin-containing solution 54: Reducing agent having SH group 56: Mixed solution of albumin-containing solution and reducing agent having SH group 60: Filtration unit 62: Filter 66: Filtered mixture 68: From mixture Waste 70 removed: Dialysis unit 72 also serving as a control unit 72: Dialyzer 74: Dialysis unit also serving as a control unit 75: Dialysate 76: Dialysate used 78: Concentration-adjusted dialysis The solution

Claims (10)

(1) Mixing an albumin-containing solution with a reducing agent having an SH group to produce a mixed solution; and (2) Dialyzing the mixed solution to contain albumin in which the ratio of reduced albumin is increased. Producing a solution,
(3) may include adjusting the concentration of albumin;
A method for producing a solution containing albumin in which the ratio of reduced albumin is increased. In (1), 10 to 90 minutes after mixing the albumin-containing solution and the reducing agent having an SH group,
The production method according to claim 1, wherein the mixture is dialyzed for 10 to 120 minutes in (2). The reducing agent having an SH group contains cysteine, and / or the molar ratio of the reducing agent having an SH group to albumin (reducing agent having an SH group / albumin) is 0.2 to 5.0. Item 3. The method according to Item 1 or 2. A solution containing albumin in which the ratio of reduced albumin is increased is stored at 20-30 ° C. for one day after dialysis, and further stored at 4 ° C. for two days for a total of three days. The ratio of reduced albumin is 50% or more. The production method according to any one of claims 1 to 3, which is at least 10% higher than the ratio of reduced albumin in the albumin-containing solution. (3) a production method including an adjustment step, and further (4) a filtration step,
The albumin-containing solution contains ascites, and (4) the filtration step removes molecules larger than albumin, and (3) the albumin concentration is increased in the adjustment step. Manufacturing method. A solution containing albumin produced by the production method according to any one of claims 1 to 5 and having an increased proportion of reduced albumin. The solution containing albumin according to claim 6, wherein the albumin-containing solution in (1) is selected from ascites. (I) A mixing unit that mixes an albumin-containing solution and a reducing agent having an SH group to produce a mixed solution; and (II) dialyzes the mixed solution to obtain albumin having an increased ratio of reduced albumin. A dialysis part for producing a solution containing,
(III) An apparatus for producing a solution containing albumin in which the proportion of reduced albumin that may include a control unit for adjusting the albumin concentration is increased. The manufacturing apparatus according to claim 8, wherein at least one of the following (i) to (iv) is satisfied.
(I) (I) After mixing the albumin-containing solution and the reducing agent having an SH group in the mixing part for 10 to 90 minutes, (II) dialyzing the mixed solution in the dialysis part;
(Ii) (II) Dialyze the mixed solution for 10 minutes to 120 minutes in the dialysis part;
(Iii) The reducing agent having an SH group contains cysteine; and (iv) The molar ratio of the reducing agent having an SH group to albumin in the albumin-containing solution (reducing agent having an SH group / albumin) is 0. .2 to 5.0. (III) A production apparatus including a control unit, and (IV) a production unit further including a filtration unit,
10. The production according to claim 8, wherein the albumin-containing solution contains ascites, and (IV) the filtration part is filtered to remove molecules larger than albumin, and (III) the albumin concentration is increased in the regulation part. apparatus.

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