JP7474516B2 - Chemical pool drugs, injection preparations, oral preparations, infusion preparations and dialysis equipment - Google Patents

Description

Application of Article 30, Paragraph 2 of the Patent Act Information regarding the present invention was published on the website at the address (https://kleinstein-biomedical-engineering-perspective-corp.jimdosite.com/) posted on May 31, 2019.

The present invention relates to chemical pool drugs, injection preparations, oral preparations, infusion preparations, and dialysis devices.

In hemodialysis and similar methods (hereinafter referred to as blood purification treatment or blood purification methods) performed as renal replacement therapy for chronic dialysis patients, various substances accumulated in the blood are removed by passing them through the semipermeable membrane of the blood purifier while performing extracorporeal circulation. There are three main principles of removal. First, there is hemodialysis, in which blood and dialysis fluid are returned to both sides of the semipermeable membrane, causing transmembrane mass transfer between the two due to the concentration gradient of the substance. Second, there is hemofiltration, in which pressure is applied to the blood to cause transmembrane filtration, and the filtrate is completely discarded while an equal amount of replacement fluid is supplied. Third, there is hemodiafiltration, which combines the first two. Such blood purification methods are being put into practical use as treatment for chronic maintenance hemodialysis. In addition, there is an example of a secondary removal principle, which is the adsorption of blood-dissolved molecules to the semipermeable membrane of the blood purifier. In some blood purification treatments, the secondary removal principle is also utilized. By continuing blood purification treatment for 4 to 6 hours per session, three times a week, the minimum necessary amount of uremic toxins can be removed and electrolytes and acid-base balance can be corrected, allowing chronic dialysis patients to survive for more than 40 years.

As a treatment for chronic renal failure, blood purification methods complement the excretory function of glomerular filtration, but cannot complement tubular reabsorption and excretion, or the endocrine function in the renal interstitium. For this reason, blood purification methods are usually administered in combination with medications such as myeloid erythroblast-stimulating hormone (erythropoietin and its derivatives), antihypertensive drugs, adsorbents that suppress the absorption of the electrolyte phosphorus from the intestinal tract, and calcium receptor agonists to suppress parathyroid function.

Furthermore, blood purification methods do not fully complement the function of eliminating reactive oxygen species and free radicals generated by the metabolism of vital activities. It is said that about 1% of oxygen molecules are converted into reactive oxygen during the breathing process without being reduced to water molecules. For this reason, highly reactive and harmful free radicals are constantly generated in the metabolic process. As a result, the chain reaction of free radicals accelerates the deterioration of various biopolymers in the bodies of chronic dialysis patients (a state of increased oxidative stress). It has been shown that blood purification treatment can also cause some reduction reactions, for example, intramolecular oxidized structures such as disulfide bonds -S-S- can be reduced to -SH. However, this reduction action is limited, and it has little complementary ability to eliminate free radicals or reduce oxidized molecules (oxidized biomolecules that are subjected to oxidation reactions in the body). Furthermore, blood purification treatments administered to chronic dialysis patients are scheduled to be repeated three times a week for 4 to 6 hours per session. As a result, there are 156 to 150 hours out of the 168 hours per week when treatment is not performed. Additionally, the interval between weekend treatment sessions and the beginning of the following week is 66-68 hours, and the interval between other sessions is about 42-44 hours. During these times when treatment is not being administered, a radical chain reaction occurs due to molecular attacks by free radicals. This causes cell damage, deteriorating the polymers that are responsible for the structure and function of the body, and ultimately worsening pathologies such as arteriosclerosis and muscle atrophy.

The number of chronic dialysis patients in Japan continues to increase, exceeding 330,000 as of 2018, and it is not uncommon for long-term survivors to last more than 40 years (see Non-Patent Document 1). As chronic dialysis patients survive for a long time, the deterioration of biopolymers, genes, and cell functions progresses due to the decreased ability to scavenge reactive oxygen species and free radicals in chronic renal failure. As a result, visible pathological conditions include muscle atrophy, motor dysfunction, arteriosclerosis, cancer, and accelerated aging. With the increase in the number of chronic dialysis patients and the aging of the population, it is feared that these problems will become even more serious in both number and quality in the future. Conventional blood purification treatments, due to defects in performance and time, are insufficient in improving the pathological condition of renal failure, which is the decreased ability to scavenge reactive oxygen species and free radicals generated during the metabolic process, and are unable to prevent long-term complications such as arteriosclerosis and muscle atrophy.

Japanese Society for Dialysis Therapy, "Illustrated Current Status of Chronic Renal Failure in Japan" (http://www.jsdt.or.jp/overview_confirm.html)

The purpose of this study is to provide a chemical pool drug that can complement the ability to eliminate reactive oxygen species (especially free radicals) and reduce oxidized biomolecules that undergo oxidation reactions in order to improve conventional blood purification treatments. In other words, the purpose of this study is to provide a chemical pool drug that can add the function of preventing the progression of chain reactions of highly chemically reactive and harmful free radicals to conventional blood purification treatments.

The present invention also aims to provide injectable, oral and infusion preparations that contain chemical pool drugs (acceptor drugs and donor drugs) used for such complementation.

Furthermore, the object is to provide a dialysis device configured to supply the infusion preparation.

These objectives are achieved by the present invention as described below in (1) to (13).

(1) An acceptor molecule capable of capturing free radicals generated in the body of a patient receiving blood purification treatment using a blood purifier, or
A chemical pool drug comprising at least one donor molecule capable of inhibiting an oxidation reaction by donating an electron as a hydrogen atom and/or a single electron to an oxidized biomolecule subjected to an oxidation reaction in the body of the patient to whom the blood purification treatment by the blood purifier is administered,
The chemical pool agent is configured to be removed from the patient's body by the blood purifier after capturing the free radicals or donating the electrons;
The chemical pool drug is administered at a chemical equivalent of 0.001 mEq/(hr kg) or more, based on the length (hr) of the treatment intermission period in the blood purification treatment and the body weight (kg) of the patient to whom the blood purification treatment is to be administered.

(2) The chemical pool drug according to (1) above, in which the total amount of chemical equivalents per blood purification treatment is 0.001 to 20 mEq/(hr·kg).

(3) A chemical pool agent as described in (2) above that is administered in an amount greater than the total amount of the chemical equivalent.

(4) A chemical pool agent according to any one of (1) to (3) above, in which the acceptor molecule after capturing the free radical and the donor molecule after donating the electron remain stable for at least 3 hours.

(5) A chemical pool agent according to any one of (1) to (4) above, which contains at least one small molecule compound having a molecular weight of 1000 D or less or a large molecule compound having a molecular weight of 20 kD or more.

(6) The chemical pool agent according to (5) above, wherein the small molecule compound contains at least one of reduced ascorbic acid and reduced glutathione.

(7) The small molecule compound is a chemical pool agent according to (5) above, which comprises a peptide consisting of 2 or 3 D-amino acid residues.

(8) The large molecule compound is a chemical pool agent according to (5) above, which includes recombinant α1-m.

(9) The recombinant α1-m is a chemical pool agent described in (8) above, which contains at least one of native recombinant α1-m or mutant recombinant α1-m.

(10) An injectable preparation comprising a chemical pool drug described in any one of (1) to (9) above.

(11) An oral preparation comprising a chemical pool drug according to any one of (1) to (9) above.

(12) An infusion preparation comprising a chemical pool drug according to any one of (1) to (9) above.

(13) A dialysis apparatus including a blood purifier, an extracorporeal blood circuit connected to the blood purifier for circulating blood, a supplying means connected to the extracorporeal blood circuit, and a dialysis fluid circuit for supplying dialysis fluid to the blood purifier,
The dialysis apparatus, wherein the supply means is configured to supply the infusion preparation according to (12) above to the extracorporeal blood circuit.

According to the present invention, it is possible to provide a chemical pool drug that can complement the ability of conventional blood purification treatments to eliminate reactive oxygen species (particularly free radicals) and to reduce oxidized biomolecules that are subjected to oxidation reactions. It is also possible to provide an injection preparation, an oral preparation, and an infusion preparation that contain a chemical pool drug (acceptor drug or donor drug) used for such complementation. Furthermore, it is possible to provide a dialysis device configured to supply the infusion preparation (for example, a rinse solution containing the drug).

FIG. 1 is an explanatory diagram of chemical pool dialysis in the treatment of chronic renal failure. FIG. 2 is an explanatory diagram showing a schematic diagram of the progress of chemical reactions of molecules of chemical pool agents in chemical pool dialysis. FIG. 1 is an explanatory diagram showing a compartment model that defines the transfer balance of chemical pool drugs between the three body compartments (extracellular fluid, cytoplasm, and mitochondria) where chemical pool dialysis is performed. FIG. 1 is an explanatory diagram showing the transition of concentration of a chemical pool drug in chemical pool dialysis in the extracellular fluid, cytoplasm, and mitochondrial fractions. FIG. 1 is an explanatory diagram showing the transition of chemical pool size when the intrinsic clearance value is different as an index of ineffective metabolism after administration of a chemical pool drug. FIG. 1 is an explanatory diagram showing a method for performing chemical pool dialysis in acute blood purification treatment. FIG. 1 is an explanatory diagram showing the relationship between the clearance value of blood purification and the removal of chemical pool drugs. FIG. 1 is an explanatory diagram showing a method of performing chemical pool dialysis in which a chemical pool drug is administered in divided doses at intermittent intervals in chronic blood purification therapy. FIG. 2 is an explanatory diagram showing an example of a molecule of a chemical pool agent. FIG. 2 is an explanatory diagram showing an example of a molecule of a chemical pool agent. FIG. 1 is a schematic diagram showing an example of the configuration of a dialysis device including a rinse solution containing a chemical pool agent.

The dialysis device, acceptor drug, donor drug, rinse solution, and method of use as a rinse solution of the present invention will be described in detail below based on the preferred embodiments shown in the attached drawings. Before proceeding with these explanations, we will briefly explain how the present invention was developed.

(History of the invention)
As mentioned above, chronic dialysis patients (patients with chronic renal failure, etc.) are in a state of increased oxidative stress. It is speculated that impaired mitochondrial function is involved in the decreased defense against oxidative stress in patients with chronic renal failure. Mitochondria are intracellular organelles essential for the metabolic activity of life. Mitochondria are responsible for the production of electron carriers NADH (nicotinamide adenine dinucleotide) and NADPH (nicotinamide adenine dinucleotide phosphate) and high-energy bond compound ATP (adenosine triphosphate) through metabolic circuits such as the citric acid cycle and the electron transport chain. NADH is used to produce ATP in mitochondria, but NADH and NADPH are also essential for the elimination of reactive oxygen species in the body in general, including mitochondria. However, when mitochondrial function declines in a state of chronic renal failure, the absolute production of NADH and NADPH, which are involved in the reduction reaction, is insufficient, resulting in insufficient elimination of reactive oxygen species, and a harmful chain reaction of free radicals is activated. In addition, the ability to reduce oxidizing substances is weakened, and oxides and peroxides accumulate.

The chain reaction of free radicals is a special oxidation-reduction reaction in which radical electrons move between molecules one after another. Depending on the characteristics of the target molecule, this chain reaction can cause cleavage of ring structures, scission, intramolecular rearrangement, glycation, carbonylation, carbamylation, nitration, methylation, oxidation, peroxidation, crosslinking, and other chemical modifications. In addition, the reaction speed of this chain reaction is fast, and most of the reaction sites are inside the cell. For this reason, there are few intermediate metabolic substances in the chain reaction that can be removed by blood purification treatment. Therefore, even if the time or frequency of blood purification treatment is increased, or drugs (various fat-soluble vitamins) are administered to remove reactive oxygen species from the blood, or continuous treatment is performed, the complementary ability to normally process the reactive oxygen species that are generated every moment is not sufficient.

In this way, in blood purification methods for treating chronic dialysis patients, it is necessary to supplement the function of eliminating reactive oxygen species, which is insufficient in current blood purification methods, and the reducing ability of oxidized biomolecules that are subjected to oxidation reactions, not only from the viewpoint of simply prolonging life but also from the viewpoint of preventing long-term complications. One of the inventions whose main purpose is to provide such supplementation is a dialysis device that can be used in combination with current blood purification methods to administer chemical pool drugs, which are different from normal drug administration. In the following, before explaining the dialysis device of the present invention, the acceptor drug and donor drug of the present invention (hereinafter sometimes referred to as "chemical pool drugs") will be explained. Note that chemical pool drugs refer to drugs used as a chemical pool, but a detailed explanation of the definition of the chemical pool will be given later. In addition, the blood purification method of the present invention that combines administration of chemical pool drugs (hereinafter referred to as "chemical pool dialysis" in the following explanation and figures) combines administration of drugs that act defensively against oxidative stress and meet certain requirements with existing blood purification methods.

(Chemical Pool Agents)
A pool is a biomedical engineering term that refers to the space in the body where a substance can be distributed. The size of a pool is usually expressed as a volume, and the amount of the substance dissolved in the pool is calculated by multiplying this volume by the concentration of the substance.

The chemical pool is a space that is constructed in the body by dissolving and distributing a drug in the body. The actual chemical pool spans three compartments: extracellular fluid (mainly consisting of plasma and interstitial fluid), cytoplasm, and mitochondria, but the volumes of each compartment are different. For example, in a 60 kg human, the volume of the extracellular fluid is 12 L, the volume of the cytoplasm is 18 L, and the volume of the mitochondria is about 6 L. In addition, the administered drug is distributed unevenly to each compartment. Therefore, the concentration of the drug in each compartment is also different. Therefore, in order to understand it in a unified manner, the size of the chemical pool present in the body is expressed in chemical equivalents rather than volumes. In other words, in the present invention, the chemical pool is the total amount of the multiple compartments in the body in which the administered drug is dissolved and distributed (i.e., extracellular fluid of about 12 L, cytoplasm of about 18 L, and mitochondria of about 6 L) collectively expressed in chemical equivalents (unit: mEq). A drug used as a chemical pool is called a chemical pool drug.

Therefore, the size of the chemical pool that is built is the chemical equivalent of the chemical pool drug that is administered. For example, if 50 mEq of a chemical pool drug is administered into the body and is not broken down in the body, a chemical pool of 50 mEq in size is thought to be built in the body. Blood purification treatment removes the chemical pool drug, reducing the size of the chemical pool.

An example of administration of chemical pool drugs is briefly explained below. As shown in "Figure 1," chemical pool drugs are repeatedly administered after each blood purification treatment. The administered chemical pool drugs form a chemical pool in the body. In this example, the size of the chemical pool, i.e., the chemical equivalent of the administered chemical pool drug, is assumed to be the chemical equivalent of reactive oxygen species that are generated during the intermittent periods of blood purification treatment and cannot be completely processed by the body's defenses. Note that in the schematic diagram of "Figure 1," the length of the intermittent periods is shown shorter than the actual length of blood purification treatment.

The chemical pool drug of the present invention is a free radical scavenger or reducing agent having the chemical reactivity shown in "Table 1". That is, the chemical pool drug is either an acceptor molecule (acceptor drug) that can capture a free radical (unpaired electron) from a radical molecule, or a donor molecule (donor drug) that can donate electrons to an oxidized biomolecule (the main body of the reduction action that is not a radical, shown by Y in Table 1) that is subjected to an oxidation reaction proceeding in the body. In the case of a donor molecule, the mode of donating electrons includes a reaction in which one electron is donated as a hydrogen atom, a reaction in which two molecules donate two electrons as two hydrogen atoms, a reaction in which one molecule donates one electron as a single electron, and a reaction in which one molecule donates one electron as a hydrogen atom and another electron as a single electron. The acceptor molecule or donor molecule before each chemical reaction is called an active molecule (unchanged body), and the acceptor molecule or donor molecule after the chemical reaction is called an inactive molecule (changed body), and the chemical pool drug before and after the reaction is distinguished. In the chemical pool, there are various ratios of active and inactive molecules, and the ratio of inactive molecules increases over time (Figure 2). In blood purification treatment, chemical pool drugs are not separated into active and inactive molecules, but are removed equally.

After administration, the molecules of chemical pool drugs become radical molecules or oxidized molecules after exerting their effect through chemical reactions in the body. If these radical molecules and oxidized molecules are not reliably removed by the next blood purification, the balance of the treatment will not have been achieved by removing free radicals or exerting a reducing effect. Therefore, chemical pool drugs must meet the following three requirements. First, the chemical equilibrium constant in "Table 1" must be greater than 1 (chemical reactivity). Second, the chemical pool drug must be stable without being decomposed in the body for a certain period of time (preferably 72 hours or more) (stability). If the chemical pool drug is decomposed in the body, the radical chain reaction will resume and the oxidation reaction of other molecules will progress, making the administration of the chemical pool drug meaningless. Third, the radical molecules or oxidized molecules, which are the reaction products of the chemical pool drug, must be removed by blood purification (removability).

Thus, the difference between the administration of chemical pool drugs of the present invention and normal drug administration is that the acceptor drug or donor drug of the present invention exists as an active molecule and an inactive molecule after administration. In other words, the inactive molecules, which are considered to have inherent harmfulness, are removed in the next dialysis, and only then can the intended effect of correcting oxidative stress and inducing homeostasis in the body to a favorable reducing state be exerted. Without removal by dialysis, the acceptor drug or donor drug of the present invention cannot exert its reducing effect. This is because free radicals are excreted from the body and a reduced state remains in the body only when the molecules of the acceptor drug or donor drug after the reaction are removed by blood purification. Therefore, the essential requirements (three essential requirements) of the chemical pool drugs of the present invention are chemical reactivity, stability after the reaction, and removability. In addition to a detailed explanation of these three requirements, further preferable requirements (non-binding, liquid-phase, accessibility, and non-metabolism) are explained below.

(Chemical Reactivity)
In order for a chemical pool agent of the present invention to be a suitable acceptor agent or donor agent, it is essential that the chemical reaction shown in "Table 1" is biased to the right side, that is, the equilibrium constant of the reaction is greater than 1. Preferably, an equilibrium constant of 1000 or more is ideal because it has a strong competitive effect in inhibiting the chain reaction of free radicals.

(Stability)
Furthermore, the chemical pool drug of the present invention requires that the acceptor drug molecules (radical type molecules) that have captured the free radicals after the reaction and the donor drug molecules (oxidized type molecules) that have been converted to an oxidized form remain stable in the body. The stable period during which the acceptor drug molecules and the donor drug molecules remain stable after the reaction is at least 3 to 9 hours (for example, the period until the detoxification reaction reaches an equilibrium state in the case of acute blood purification treatment), and preferably 48 to 72 hours (for example, the interim period until the next blood purification in the case of chronic blood purification treatment). In addition, when the equilibrium constant of the chemical reaction shown in "Table 1" is 1000 or more, it contributes to the continued stable existence of the acceptor drug molecules that have captured the free radicals after the reaction and the donor drug molecules that have been converted to an oxidized form.

(Removability)
Since the chemical pool drugs are assumed to be removed by blood purification treatment, it is necessary that the dialysability or filterability by the blood purifier is guaranteed. When hemodialysis is selected as the blood purification method constituting the chemical pool dialysis, the molecular weight of the chemical pool drug is preferably 1000 D or less, more preferably 500 D or less. The molecular weight of the monomer of the drug that becomes a dimer after reaction (for example, the donor drug "Table 1" that forms a dimer after reaction) is preferably 500 D or less. This is because the removal efficiency by blood purification decreases with increasing molecular weight. In addition, when hemodiafiltration, hemofiltration, or plasma exchange, which are excellent in removing large molecules, are selected instead of hemodialysis as the blood purification method constituting the chemical pool dialysis, the requirement of the above removal ability is relaxed because it is possible to remove large molecules with a molecular weight in the range of 20 to 40 kD.

In addition, in order to efficiently remove the chemical pool drug after the reaction and to create a new chemical pool by administering a new chemical pool drug, it is preferable to set the clearance value, which indicates the removal performance of the blood purifier (removal performance of the artificial kidney), to 200 mL/min or more so that the chemical pool drug can easily pass through the dialysis membrane, as described below.

(Non-bonding)
In addition, it is preferable that the chemical pool drug of the present invention does not bind to protein molecules with large molecular weight, cell membranes, or extracellular matrices, because the molecules of the chemical pool drug bound to these become difficult to remove by blood purification.

Specifically, it is preferable that the molecules of chemical pool drugs do not bind to circulating medium molecules with a molecular weight of 10 kD or more that exist in the body or tissue-constructing macromolecules. The efficiency of removal of the former by blood purification decreases as the molecular weight increases, and removal of the latter is virtually impossible.

(Liquid-compatible)
As mentioned above, the chemical pool drug of the present invention must be removed with high efficiency by blood purification. Therefore, the molecules of the chemical pool drug are preferably molecules that are not lipophilic (hydrophobic) or amphiphilic, and are dissolved in the cell membrane or the membrane of the intracellular organelle, and are incorporated into these membranes. This is because the molecules of the chemical pool drug that are incorporated into the cell membrane and solidified therein cannot be removed by blood purification. For example, vitamin E is known to be a strong radical scavenger, but is lipophilic. For this reason, vitamin E is mainly incorporated into the cell membrane, and is difficult to remove by blood purification. Drugs consisting of such lipophilic molecules cannot be used as the chemical pool drug of the present invention.

(Reachability)
In the body, reactive oxygen species can be generated in all fractions, including the cytoplasm and plasma, but most are generated in the mitochondria. For this reason, mitochondria are most susceptible to damage by free radicals. Mitochondria also produce NADH, the most important reducing substance. The products of the chain reaction of free radicals that are not processed in the mitochondria leak into the cytoplasm, and the products of the chain reaction that are not processed in the cytoplasm also escape outside the cell. For this reason, the most important target of the chemical pool drug of the present invention is mitochondria. Therefore, it is preferable that the chemical pool drug of the present invention penetrates the outer mitochondrial membrane and reaches the inside of the mitochondria (including the matrix that penetrates the inner membrane). For each chemical pool drug, a drug that reaches the inside of the mitochondria by 10% or more of the administered content (chemical equivalent) is considered to satisfy the requirement of reachability.

That is, it is desirable that the chemical pool drug of the present invention has a high degree of reach or distribution to mitochondria, which are the generating site of reactive oxygen species. Medically, this reach is related to the permeability coefficient of the outer and inner mitochondrial membranes. The higher the permeability coefficient of a chemical pool drug, the smaller the chemical equivalent amount that can be administered to produce an effect. An acceptor or donor drug that satisfies at least the three essential requirements and also satisfies the requirement of mitochondrial reachability is called a "mitochondrial reachable chemical pool drug." An acceptor or donor drug that satisfies at least the three essential requirements but does not satisfy the requirement of mitochondrial reachability is called a "non-mitochondrial reachable chemical pool drug." The non-mitochondrial reachable chemical pool drug includes one that reaches the cytoplasm (first non-mitochondrial reachable chemical pool drug) and one that does not reach the cytoplasm (second non-mitochondrial reachable chemical pool drug), but both can be used as the chemical pool drug of the present invention.

(non-metabolizable)
When the chemical pool drug of the present invention is administered to a chronic dialysis patient, the chemical pool drug constituting the chemical pool is required to exert its effect during the 48-72 hour treatment intermittent period. For this reason, it is preferable that the chemical pool drug can exist without being decomposed irrelevant to the intended chemical reaction. Decomposition irrelevant to the intended chemical reaction is called ineffective metabolism, and if ineffective metabolism is present, the dosage of the chemical pool drug must be increased. For example, when a certain patient is to be administered 50 mEq of a certain drug, if the ineffective metabolic rate of this chemical pool drug is 60% in 72 hours, the initial 50 mEq is reduced to 20 mEq. When administering such a chemical pool drug, the initial dosage must be increased to 50/(1-0.6) = 125 mEq in order to ensure the expected strength of action. The lower the ineffective metabolic rate of the chemical pool drug of the present invention, the smaller the dosage can be. "Figure 5" is an explanatory diagram showing the magnitude of the ineffective metabolic rate expressed as an index of intrinsic clearance and comparing the effect on the size of the chemical pool. Specifically, in Fig. 5, the reference numeral 6 indicates the change in the size (mEq) of the chemical pool when the intrinsic clearance value is 1 mL/min, the reference numeral 7 indicates the change in the size (mEq) of the chemical pool when the intrinsic clearance value is 2 mL/min, and the reference numeral 10 indicates the change in the size (mEq) of the chemical pool when the intrinsic clearance value is 12 mL/min. Meanwhile, the reference numeral 11 in Fig. 5 indicates the change in the amount (mEq) of the chemical pool drug present in the mitochondria when the intrinsic clearance value is 1 mL/min, the reference numeral 12 indicates the change in the amount (mEq) of the chemical pool drug present in the mitochondria when the intrinsic clearance value is 2 mL/min, and the reference numeral 15 indicates the change in the amount (mEq) of the chemical pool drug present in the mitochondria when the intrinsic clearance value is 12 mL/min. As is clear from Fig. 5, the smaller the intrinsic clearance of the living body for the chemical pool drug, the more chemical pool drug continues to exist in the body (mitochondria), so it is preferable that the intrinsic clearance of the living body for the chemical pool drug is small.

Candidates for the chemical pool drugs of the present invention, which may be acceptor drugs and donor drugs, include peptides consisting of 2-3 residues of D-amino acids. More specifically, dipeptides consisting of 2 residues of D-amino acids (e.g., D-lysyl tyrosine, D-tyrosyl lysine, D-tyrosyl cysteine, D-tryptophanyl tyrosine, D-glycyl tyrosine, etc.) and tripeptides consisting of 3 residues of D-amino acids (e.g., D-lysyl tyrosyl cysteine, D-tyrosyl tryptophanyl lysine, etc.) are exemplified as candidates for the acceptor drug and donor drug. Among these, dipeptides or tripeptides consisting of 2-3 residues of D-amino acids as shown in Table 2 below are preferred as chemical pool drugs.

As an example of the acceptor drug of the present invention, 2-(α-D-glucopyranosyl) (methyl)methyl-2,5,7,8-tetramethylchroman-6-ol shown in FIG. 9 can be given. This methyl TMG shown in FIG. 9 is designed to prevent hydrolysis by α-glycosidase in the body after administration as a chemical pool drug by modifying the methyl group next to the ether bond between glucose and the chroman ring with a methyl group, so that the active center of the enzyme is not close due to steric hindrance. TMG without this modified methyl group is easily hydrolyzed and cannot stably survive in the body. Furthermore, 2-(α-D-glucopyranosyl) (isopropyl)methyl-2,5,7,8-tetramethylchroman-6-ol shown in FIG. 10 can also be given as an example of the acceptor drug of the present invention. Alkyl group-modified TMG (TMG with the center modified with an alkyl group) such as methyl TMG in FIG. 9 and isopropyl TMG in FIG. 10 are suitable as acceptor drugs of the present invention.

However, the chemical pool agent of the present invention is not limited to the above description, and for example, water-soluble vitamin E modified molecules (acceptor agent) and glutathione (donor agent) can also be used. In addition, reduced ascorbic acid can be a candidate for the chemical pool agent of the present invention from the viewpoint of chemical reactivity, but it is necessary to consider that it has a considerable amount of ineffective metabolism and that its radical form releases radicals next. In other words, reduced ascorbic acid alone does not meet the stability requirements of a chemical pool agent. Therefore, it is possible to use a donor agent (e.g., glutathione) in combination with reduced ascorbic acid to compensate for the shortcomings in the stability of reduced ascorbic acid. This makes it possible for reduced ascorbic acid to be used as an agent that becomes the chemical pool of the present invention. In other words, when multiple chemical agent species, including chemical agent species that do not satisfy the three essential requirements, as a whole satisfy the three essential requirements, such multiple chemical agent species as a whole can be considered as the chemical pool agent of the present invention. It is also necessary to consider that reduced ascorbic acid does not reach mitochondria. It is also necessary to consider that glutathione has a considerable amount of ineffective metabolism, but it reaches mitochondria. Therefore, from the viewpoint of supplementing mitochondrial accessibility, the combined use of reduced ascorbic acid and glutathione is preferable. Thus, reduced ascorbic acid is not suitable as a chemical pool drug by itself because it does not reach mitochondria and is not stable enough, but on the other hand, it can be said to be useful as an adjuvant for chemical pool drugs because of its extremely high radical scavenging ability. For example, reduced ascorbic acid can scavenge radicals that have entered solid-phase membrane lipids and transfer them to chemical pool drugs (e.g., glutathione). Generally, receiving electrons means that a molecule is reduced, but when reduced ascorbic acid receives one radical, it loses two hydrogen atoms (H), which consist of one proton and one electron, and becomes an oxidized form.

Here, we will explain the amount (mEq) of the chemical pool in each of the three fractions, the extracellular fluid, the cytoplasm, and the mitochondria, and then discuss the specific chemical equivalents for each application of the chemical pool drug of the present invention. "Figure 3" is a diagram of a compartment model showing the balance of movement of the chemical pool drug in and outside the three fractions of the chemical pool, the extracellular fluid, the cytoplasm, and the mitochondria. The chemical pool drug changes its active molecule to an inactive molecule through chemical reactions. For the chemical pool drug administered to the body, the conversion rate from active to inactive molecules is set as Qe mEq/min in the extracellular fluid, Qc mEq/min in the cytoplasm, and Qm mEq/min in the mitochondria. In each fraction, the active molecule decreases at these conversion rates, and the inactive molecule increases at these conversion rates. The intermittent period of blood purification treatment can be two days (about 43 hours) or three days (about 67 hours), so the dose of the chemical pool drug at the beginning of each period is set as A1, A2 (A1<A2) mEq. The permeation rate determined by the concentration difference at both ends of the cell membrane that separates the extracellular fluid from the cytoplasm is Kc mL/min. The permeation rate determined by the concentration difference at both ends of the mitochondrial outer membrane that separates the cytoplasm from the mitochondria is Km mL/min. The clearance that represents the removal performance of the artificial kidney is Kd mL/min. The metabolism of the molecules of the chemical pool drug in the living body, which is unrelated to the conversion of the chemical pool drug from an active molecule to an inactive molecule and removal by blood purification, is called ineffective metabolism. The index representing this ineffective metabolism is set to intrinsic clearance Ki mL/min, and it is assumed that the drug is ineffectively metabolized from the cytoplasm. 40% of the body weight of a patient undergoing chemical pool dialysis (chronic dialysis patient) is considered to be intracellular fluid, of which 75% is considered to be the cytoplasmic volume Vc and 25% is considered to be the mitochondrial volume Vm. The extracellular fluid of a chronic dialysis patient is 20% of the body weight under normal conditions, but due to oliguria or anuria, it increases during intermittent periods, and the increase is corrected by water removal during blood purification treatment. If we consider that the body fluid increases at a constant rate during the intermittent period, the extracellular fluid volume can be expressed as a function of time v(t). The concentrations of the three compartments for active and inactive molecules are expressed as functions of time. The concentration trends over time of the extracellular fluid, cytoplasm, and mitochondria during the intermittent period of blood purification treatment, for example, the intermittent period starting after blood purification treatment at the beginning of the week, are calculated using the numerical solution of the simultaneous differential equations in "Equation 1." During blood purification treatment, for example, during blood purification treatment in the middle of the week, the concentration trends over time of the extracellular fluid, cytoplasm, and mitochondria are calculated using the numerical solution of the simultaneous differential equations in "Equation 2." A two-week concentration profile is obtained by substituting the final value of the previous interval for the initial value of the next interval.

Figure 4 shows the results of multiplying the concentration profile thus obtained by the volume of each fraction. In Figure 4, the transition of the abundance (mEq) of the chemical pool drug is shown for each chemical pool fraction. The combined abundance of the active molecules of the chemical pool drug and the inactive molecules after the reaction is shown here. The arrival of the chemical pool drug at the deep pool, the mitochondria, and the reverse mass transfer, the mitochondrial purification, are the slowest. If the removal efficiency of the chemical pool drug is low, the concentration transition curve shifts upward.

In the formula, α[t] is a notation that indicates that α is a function of the variable t. D[β[t] v[t], t] means to differentiate β[t] v[t] with respect to t. The same goes for the others. Note that α[t] represents the concentration (mEq/mL) of the active molecule in the extracellular fluid. γ[t] represents the concentration (mEq/mL) of the active molecule in the cytoplasm. ε[t] represents the concentration (mEq/mL) of the active molecule in the mitochondria. e[t] represents the amount (mEq) of the active molecule in the extracellular fluid. β[t] represents the concentration (mEq/mL) of the inactive molecule in the extracellular fluid. δ[t] represents the concentration (mEq/mL) of the inactive molecule in the cytoplasm. ζ[t] represents the concentration (mEq/mL) of the inactive molecule in the mitochondria. f[t] represents the amount of inactive molecule present in the extracellular fluid (mEq/mL).

Let the calculated values be α[2580]=α2580, γ[2580]=γ2580, ε[2580]=ε2580, e[2580]=e 2580, β[2580]=β2580, δ[2580]=δ2580, ζ[2580]=ζ2580, and f[2580]=f 2580.These values (which are actually numbers) are substituted for the initial values of the next time interval ("Math 2").

The chemical pool drug of the present invention is mainly intended to be administered to chronic dialysis patients (in combination with chronic blood purification treatment), but is also intended to be administered to patients with acute renal failure, patients with drug or poison poisoning, patients with severe infections, patients with burns, patients with post-ischemic reperfusion injury, patients with severe pregnancy toxemia, patients with chronic skin ulcers, patients with cerebral hemorrhage and ventricular rupture, patients with severe hemolytic diseases, etc. (in combination with acute blood purification treatment). The case of administering a chemical pool drug to a chronic dialysis patient will be described. First, at the end of a treatment session of a normal blood purification method performed on a schedule of three times a week, the required amount of the chemical pool drug is administered to the chronic dialysis patient. Then, during the treatment interval until the next blood purification treatment, the molecules of the chemical pool drug that form the chemical pool capture free radicals through a chemical reaction or exhibit reducing properties. Next, the radical molecules and oxidized molecules of the chemical pool drug after the reaction exist stably without causing the next chemical reaction. Finally, the molecules of the chemical pool drug that exist stably as radicals or oxidized forms are removed by the next blood purification treatment. This allows harmful free radicals and oxidized forms (oxidized molecules) to be excreted from the body. The cycle then repeats, with the chemical pool drug administered again after blood purification treatment.

When treating a patient with chronic renal failure according to the present invention, the size of the chemical pool to be administered, i.e., the total amount of chemical equivalents of the chemical pool drug, is preferably 0.001 to 0.05 mEq/(hr·kg), and more preferably 0.01 to 0.05 mEq/(hr·kg), using the body weight (kg) of the patient with chronic renal failure and the length of the treatment interval (hr). This is a value that assumes that, taking into account the results of the above formula, the chemical equivalents of the reactive oxygen species produced in the body that can escape endogenous elimination and become the starting point of harmful chain reactions are neutralized. Thus, the chemical pool drug is administered in large quantities. For this reason, it is a natural requirement for the chemical pool drug to be non-toxic to the human body even when administered in large quantities into the body. It is misunderstood that the removal of the chemical pool drug has been almost completed based on the transition of the blood concentration. However, as can be seen from "Figure 4", it is actually believed that a certain amount of the chemical pool drug remains in the cytoplasm and mitochondria even at the end of blood purification, and a steady state is reached. For this reason, it is preferable that chemical pool agents are not cytotoxic upon chronic exposure.

As another application of the chemical pool drug, the administration of the chemical pool drug to patients with acute renal failure, patients with drug or poison poisoning, patients with severe infections, patients with burns, patients with post-ischemic reperfusion injury, patients with severe pregnancy toxemia, patients with chronic skin ulcers, patients with cerebral hemorrhage and ventricular rupture, and patients with severe hemolytic disease will be described. First, the chemical pool drug is administered to the patient, and an equilibration period of several hours to several days is allowed. After that, blood purification treatment is performed once or several times to remove the captured free radicals and oxidized molecules from the body. The chemical pool drug of the present invention can also be used in such treatments. One course of this treatment is shown in "Figure 6". The size of the chemical pool is divided into the white area (chemical equivalent of the active molecules remaining in the chemical pool drug) and the filled area (chemical equivalent of the inactive molecules after the chemical reaction). The purpose of treatment here is to save a life, unlike the prevention of complications in chronic renal failure, so the amount of the chemical pool drug administered is larger than in the case of chronic renal failure. The above course is repeated several times as necessary.

When the chemical pool drug of the present invention is administered to patients with acute renal failure, patients with drug or poison poisoning, patients with severe infections, patients with burns, patients with post-ischemic reperfusion injury, patients with severe pregnancy toxemia, patients with chronic skin ulcers, patients with cerebral hemorrhage and ventricular rupture, and patients with severe hemolytic diseases, the size of the administered chemical pool, i.e., the total amount of chemical equivalents of the chemical pool drug, is preferably 0.1 to 20 mEq/(hr·kg) using the patient's body weight (kg) and the length of the treatment interval (hr). This is greater than the amount administered when treating patients with chronic renal failure. The reason for this is that it is considered that the invasion of the body by diseases and adverse events increases the production of reactive oxygen species.

As the chemical pool drug of the present invention, it is preferable to use a combination of multiple drug species administered as the chemical pool drug. For example, it is preferable to use a combination of an acceptor drug that captures free radicals and a donor drug that is an H donor as the chemical pool drug. It is more preferable that the drug species administered as the chemical pool drug contains at least one mitochondrial-reaching chemical pool drug such as glutathione. This makes it possible to block the chain reaction of free radicals at multiple stages. That is, it is possible to effectively reduce oxidative stress in the extracellular fluid, cytoplasm, and mitochondria. As a result, it is possible to improve the blocking rate of harmful chain reactions in the living body. In addition, when the chemical pool drug is composed of multiple drug species including at least a mitochondrial-reaching chemical pool drug, it is preferable that the content (chemical equivalent) of the mitochondrial-reaching chemical pool drug is higher than the total content of the other drug species. In other words, it is preferable that the chemical pool drug mainly contains a mitochondrial-reaching chemical pool drug.

(Method of administering chemical pool agents)
Chemical pool drugs are usually administered intravenously to ensure proper administration, since the dose is usually several tens to several hundreds of mEq. For example, in an infusion preparation in which 100 mEq of a chemical pool drug is dissolved in 200 mL of water, the osmolality ratio of the infusion preparation to physiological saline is 100x(1000/200)/308=1.6. Infusion preparations of this type are easy to administer intravenously. If an infusion preparation is prepared by dissolving the drug in 10 mL of water, the osmolality ratio of the infusion preparation to physiological saline is 16, which is quite high. Chronic dialysis patients have internal shunts and indwelling catheters to maintain the blood flow of the extracorporeal circulation. For this reason, it can be said that chronic dialysis patients can tolerate the infusion of infusion preparations with higher osmolality than that of normal peripheral skin veins (osmolality ratio is limited to 3). However, it can be said that side effects are generally less likely to occur when drugs are administered slowly as infusion preparations with a larger volume than when administered as a one-shot intravenous injection.

As a method of administration of the chemical pool drug of the present invention, administration by oral administration is also considered. In this case, taking into consideration the burden of oral administration on the patient, the drug is administered once or several times a day in divided doses during intermittent blood purification treatment (Figure 8). When taken orally, the chemical equivalent of the chemical pool drug that actually enters the body and builds the chemical pool decreases due to the effects of decomposition in the intestinal tract, the absorption rate in the intestinal tract, etc. Taking this into consideration, it is necessary to adjust the chemical equivalent of the chemical pool drug taken orally.

As described above, intravenous administration at the end of blood purification is suitable as a method for administering the chemical pool drug of the present invention. By administering it all at once, the concentration gradient becomes large between the extracellular fluid at the surface and the cytoplasm at the deep layer, and also between the mitochondria at the deep layer. As a result, the driving force for the chemical pool drug to reach the deep layer due to diffusion also becomes large. In addition, it takes time for the chemical pool drug to reach the deep layer. For this reason, administering the chemical pool drug at the beginning of the treatment intermission and using the entire intermission as the reaction time is also advantageous for the manifestation of the therapeutic effect.

In particular, when administering the chemical pool drug of the present invention to a chronic dialysis patient, the first choice is to administer a chemical equivalent of the chemical pool drug required to neutralize reactive oxygen species generated during intermittent periods intravenously all at once at the end of blood purification. This is because the amount of the chemical pool drug that reaches the deep mitochondria to which it is desired to be distributed increases with the concentration gradient and the time available for transfer.

(Rinse solution)
The rinse liquid is used to push back the blood remaining in the extracorporeal blood circuit to the internal circulation (inside the blood vessels) after the end of blood purification. The rinse liquid of the present invention is configured to contain the above-mentioned chemical pool drug and a solvent such as water or physiological saline. When an infusion preparation is selected as the dosage form of the chemical pool drug as described above, the infusion preparation can also be used as the rinse liquid. This can reduce the volume load of the chemical pool drug on the patient. When the chemical pool drug is also used as the rinse liquid, the volume of the rinse liquid (infusion preparation of the chemical pool drug) is preferably 200 mL to 1000 mL, more preferably 200 to 400 mL. The rinse liquid is preferably administered in its entirety over several minutes or more. The osmotic pressure ratio of the rinse liquid to physiological saline is preferably 0.5 to 8, more preferably 0.5 to 5.

Furthermore, the rinse solution may contain an auxiliary agent such as an L-type amino acid. When an L-type amino acid is used as an auxiliary agent, the content of the L-type amino acid is preferably 1 g to 20 g in total, and more preferably 3 g to 15 g in total. During blood purification treatment, leakage of amino acids from muscles is inevitable. Therefore, physiological amino acids administered immediately after blood purification treatment are re-absorbed into muscles and exert the effect of suppressing the progression of muscle atrophy. Therefore, when the rinse solution contains an L-type amino acid, a synergistic effect with the prevention of muscle atrophy through the mitochondrial protection function of the chemical pool drug of the present invention can be expected. In the present invention, as a result of seeking highly efficient removal of the molecules (small molecules) of the chemical pool drug, the loss of amino acids in the body fluids also increases accordingly, and an increase in the amount of amino acids leaking out of the muscles is inevitable. In order to alleviate this adverse effect, the co-administration of an L-type amino acid is also effective.

When the rinse solution contains an L-amino acid as an auxiliary agent, the osmotic pressure ratio of the rinse solution to physiological saline is preferably 2 to 8. In this range of osmotic pressure ratios, slow administration (recovery operation) over several minutes or more is particularly necessary. Below, the dialysis device of the present invention, which can supply chemical pool agents to the body as a rinse solution, will be described in detail based on the preferred embodiment shown in the attached drawings.

(Dialysis machine)
Fig. 11 is a schematic diagram showing an example of the configuration of a dialysis device equipped with a rinse solution containing a chemical pool drug. The dialysis device 100 of the present invention enables hemodialysis, hemofiltration, hemodiafiltration, and plasma exchange, which are already used in clinical settings, but the following description will be given taking hemodialysis as an example. That is, the dialysis device 100 has the same basic configuration as a fully automatic dialysis device, and can perform blood purification treatment using a general automatic dialysis device.

The dialysis device 100 of the present invention basically comprises a blood purifier (dialyzer) D, a dialysate circuit 200 and an extracorporeal blood circuit (blood circuit) 300 connected to the blood purifier D, and a replacement fluid supply circuit 400 connected to the blood circuit 300 so as to branch off on the arterial side of the patient's blood vessels.

The blood purifier D has multiple straw-shaped capillaries D1. Each capillary D1 is formed of a semipermeable membrane (dialysis membrane), which has multiple holes. The blood purifier D is configured so that blood flows inside each capillary D1 while dialysis fluid flows outside each capillary D1. This allows the blood and dialysis fluid to come into contact with each other via the semipermeable membrane. This contact causes harmful substances in the blood to transfer to the dialysis fluid.

In the blood purifier D, it is necessary to remove radical molecules and oxidized molecules, which are reaction products of chemical pool drugs, from the body with high efficiency. For this reason, it is preferable that the blood purifier D has a clearance value of 150 mL/min or more, preferably 200 mL/min, as the removal performance for small molecules with a molecular weight of 1000 D or less (see "Figure 7"). In Figure 7, reference numeral 16 indicates the change in chemical pool size (mEq) when the clearance value of the blood purifier D is 100 mL/min, which is the removal performance of the blood purifier D, reference numeral 17 indicates the change in chemical pool size (mEq) when the clearance value of the blood purifier D is 150 mL/min, and reference numeral 20 indicates the change in chemical pool size (mEq) when the clearance value of the blood purifier D is 300 mL/min. As is clear from Figure 7, in the range of the clearance value of the blood purifier D from 100 mL/min to 300 mL/min, the higher the clearance value of the blood purifier D, the more chemical pool drugs are removed. For this reason, the clearance value of blood purifier D, which is the removal performance for small molecules with a molecular weight of 1000D or less, is preferably 100mL/min to 300mL/min, more preferably 150mL/min to 300mL/min, and even more preferably 200mL/min to 300mL/min. In the chemical pool, purification of the extracellular fluid at the surface takes place first, followed by purification of the cytoplasm deep inside and mitochondria even deeper inside. Purification of the deep pool is time-limited. The typical blood purification treatment time in clinical settings is 4 to 5 hours, but the longer the treatment time, the higher the degree of purification of the chemical pool.

The dialysis fluid circuit 200 includes multiple pumps that circulate the dialysis fluid and supply it to the blood purifier D. The multiple pumps are composed of a dialysis fluid delivery pump P1, a dialysis fluid drainage pump P2, and a third dialysis fluid pump P3. The dialysis fluid delivery pump P1 and the dialysis fluid drainage pump P2 have the function of always circulating the dialysis fluid at an equal delivery speed. In other words, the dialysis fluid delivery pump P1 and the dialysis fluid drainage pump P2 have the function of forming an equal flow rate system. On the other hand, the third dialysis fluid pump P3 is configured to intertwine with the interior of the equal flow rate system. Specifically, the third dialysis fluid pump P3 is configured to rotate in both forward and reverse directions, and can drain the dialysis fluid in the forward direction or inject the dialysis fluid in the reverse direction.

The blood circuit 300 is composed of an arterial blood circuit 310 and a venous blood circuit 320 via a blood purifier D. The arterial blood circuit 310 is provided with an electromagnetic opening/closing clamp C2 and a blood pump P4 that rotates in both directions. By rotating the blood pump P4 in the forward direction, blood taken from the artery is supplied to the blood purifier D, comes into contact with the dialysis fluid, and then returns to the vein through the venous blood circuit 320. When blood is flowing in this manner, the electromagnetic opening/closing clamp C2 is in an open state. On the other hand, when stopping such blood flow, the electromagnetic opening/closing clamp C2 is in a closed state. When supplying replacement fluid, which will be described later, the electromagnetic opening/closing clamp C2 is in a closed state. In the arterial blood circuit 310, a replacement fluid supply circuit 400 is connected between the electromagnetic opening/closing clamp C2 and the blood pump P4.

The replacement fluid supply circuit 400 is provided with replacement fluid. The replacement fluid includes an electrolyte solution such as pure water or saline, and in this embodiment, further includes a chemical pool agent. The replacement fluid supply circuit 400 is also provided with an electromagnetic opening/closing clamp C1 that can stop the supply of replacement fluid. The replacement fluid is supplied to the blood circuit 300 by opening the electromagnetic opening/closing clamp C1 and closing the electromagnetic opening/closing clamp C2 and rotating the blood pump P4 forward.

The dialysis device 100 of the present invention has an electromagnetic opening/closing clamp C1 and an electromagnetic opening/closing clamp C2 that can be electromagnetically opened and closed, a blood pump P4 that can rotate in both directions, and a third dialysis fluid pump P3 that can rotate in both directions. This allows the same amount of water to be removed from the blood circuit 300 or the same amount of fluid to be replaced in the blood circuit 300 through the semipermeable membrane of the blood purifier D. The dialysis device 100 of the present invention is characterized in that each of the dialysis fluid circuit 200 and the blood circuit 300 has at least one pump that can rotate in both directions. In addition to the basic configuration described above, the dialysis device 100 can also have various elements (e.g., a control unit and a sensor). The control unit can programmatically control the opening and closing of the electromagnetic opening/closing clamp C1 and the electromagnetic opening/closing clamp C2, and the rotation direction and speed of the third dialysis fluid pump P3 and the blood pump P4 to any value at any time.

(Method of using a solution containing chemical pool cleaners as a rinse solution)
11, a method of using the replacement fluid containing the chemical pool drug of the present invention as a rinsing fluid R for blood recovery in the extracorporeal blood circuit 300 of the dialysis device 100 will be described below. Two specific examples are shown below.

(Specific Example 1)
As a replacement fluid, 200 to 300 mL of rinse fluid R, which is an aqueous solution containing the chemical pool agent of the present invention, is connected to the replacement fluid supply circuit 400, which is a branch of the blood circuit 300. During hemodialysis, the electromagnetic opening/closing clamp C1 is closed, the electromagnetic opening/closing clamp C2 is open, the blood pump P4 rotates forward (from s to v), and the third dialysis fluid pump P3 operates in the forward direction (water removal). The above is a pre-recovery process. Upon completion of blood purification treatment (usually the time when the specified treatment time has expired and the specified amount of water removed has been reached), the following series of steps are operated. 1) The dialysis fluid delivery pump P1 and the dialysis fluid drainage pump P2 are stopped. 2) The electromagnetic opening/closing clamp C1 is opened, the electromagnetic opening/closing clamp C2 is closed, and the blood pump P4 is rotated forward at a speed of 10 to 100 mL/min, and the entire amount of rinse fluid R is injected into the blood circuit 300 from t. This injection amount is the value obtained by multiplying the flow rate of the blood pump P4 by the operation time. After that, the forward rotation of the blood pump P4 is stopped. At this point, the blood from t to v is returned to the body, and the same part is replaced with the rinse solution R (i.e., an aqueous solution containing a chemical pool drug). 3) The electromagnetic opening/closing clamp C1 is closed, the electromagnetic opening/closing clamp C2 is opened, and the third dialysis fluid pump P3 is operated in the reverse direction (fluid replacement direction) at a speed of 10 to 100 mL/min, and the blood pump P4 is operated in the reverse direction at an arbitrary speed lower than that of the third dialysis fluid pump P3. As a result, the dialysis fluid is backfiltered through the semipermeable membrane and injected into the blood circuit 300. Furthermore, the dialysis fluid is distributed in an arbitrary ratio in two directions, s and v. As a result, the blood remaining in the part from s to t of the blood circuit 300 is returned from s, and the rinse solution R (aqueous solution containing a chemical pool drug) remaining in the part from t to v is replaced with the dialysis fluid and injected into the body. As a result, the administration of the chemical pool drug and the blood return process are completed at the same time.

(Specific Example 2)
A blood return process using an aqueous solution containing chemical pool agents by another program control is shown. The pre-recovery process is the same as that of the specific example 1. 1) The dialysis fluid pump P1 and the dialysis fluid drain pump P2 are stopped. 2) With the electromagnetic open/close clamp C1 in the closed state and the electromagnetic open/close clamp C2 in the open state, the third dialysis fluid pump P3 is operated in the reverse direction (fluid replacement direction) at a speed of 10 to 100 mL/min, and the blood pump P4 is operated in the reverse direction at the same speed. As a result, the dialysis fluid equivalent to the volume of the blood circuit 300 from s to u is backfiltered through the semipermeable membrane and injected into the blood circuit 300 side. As a result, the blood in the portion from s to u of the blood circuit 300 is returned from s first. 3) The third dialysis fluid pump P3 is stopped, the electromagnetic open/close clamp C1 is opened, the electromagnetic open/close clamp C2 is closed, and the blood pump P4 is operated in the forward direction at a speed of 10 to 100 mL/min. As a result, the entire amount of the rinse fluid R (aqueous solution containing chemical pool agents) is injected. As a result, blood remaining in the blood circuit 300 from u to v is returned from v. This infusion amount is the value obtained by multiplying the flow rate of the blood pump P4 by the operation time. 4) The electromagnetic opening and closing clamp C1 is closed, the blood pump P4 is stopped, and the third dialysis fluid pump P3 is operated in the reverse direction (fluid replacement direction) at a speed of 10 to 100 mL/min. As a result, the aqueous solution containing the chemical pool drug remaining in the part from u to v of the blood circuit 300 is replaced with dialysis fluid of the same volume (or 0.5 to 1.5 times the volume) of the blood circuit 300 in the same part, and is infused into the body. The replacement amount by backfiltration is the value obtained by multiplying the flow rate of P3 by the operation time.

In Example 2, blood is returned from the portion s to t of the blood circuit 300 earlier than in Example 1, so there is less risk of blood clotting in that portion during the recovery process. However, there is a risk that a small amount of chemical pool drug solution may remain in the portion t to u of the blood circuit 300.

As explained above, the dialysis device of the present invention enables chemical pool dialysis (a blood purification method in which the administration of the chemical pool drug of the present invention is used in combination). The chemical pool drug administered by chemical pool dialysis preferably penetrates the cell membrane from the shallow extracellular fluid to the deep cytoplasm and then to the mitochondria in the deepest layer. By performing chemical pool dialysis using the dialysis device of the present invention, a chemical pool can be formed in these three compartments. The molecules of the chemical pool drug dissolved in this chemical pool capture free radicals in mitochondria, which are the source of reactive oxygen species, and also exert reducing properties against reactive oxygen species, thereby reducing oxygen damage. Therefore, the present invention contributes to the recovery of mitochondrial function damaged in patients with chronic renal failure. The recovery of mitochondrial function restores the production of NADH, an electron carrier, and ATP, a compound with high energy bonds. Therefore, the recovery of mitochondrial function improves the endogenous defense against reactive oxygen species and also improves the energy supply to cells. In this way, it is possible to prevent the reduced motor ability and muscle atrophy of skeletal muscles in chronic dialysis patients, and to prevent symptoms of muscle atrophy, muscle weakness, and motor dysfunction, called sarcopenia, locomotive syndrome, or frailty, which progress as a result of chronic renal failure pathology and the aging process working together. Heart failure, an important cause of death in chronic renal failure patients, is also related to myocardial damage more than ischemia, and the present invention is expected to improve heart failure. In addition, preventing the progression of the chain reaction of free radicals in the mitochondria, which is at the very top of the body, reduces oxidative stress in the cytoplasm, which is downstream.

Molecules of the chemical pool drug distributed in the cytoplasm capture free radicals that are the products of chain reactions of free radicals that have leaked from mitochondria into the cytoplasm and reduce these molecules. As a result, deterioration of intracellular organelles, cytoskeleton, and cell membranes can be prevented.

The cytoplasm is a site where a variety of reaction products are produced in addition to the radical chain, radical amplification, and peroxide generation of the initial free radical chain reaction. For example, reactive oxygen species attack protein molecules, ultimately causing molecular degradation of the peroxidation type and carbonylation type, called AOPP (advanced oxidation protein products). In addition, the hydroxyl radical (·OH) at the beginning of the chain reaction and the peroxynitrite (ONOO·) generated afterwards promote the production of carboxymethyllysine (CML), which is an advanced glycation end product (AGE). CML deteriorates biopolymers by cross-linking. In addition, alkoxyl radicals (LO·) and alkylperoxy radicals (LOO·) produced by the attack of lipids by free radicals produce hydroxynonenal (4-HNE) as a highly reactive activated carbonyl compound. 4-HNE exhibits activity against a variety of cells, and can cause cell death when it acts on DNA or mitochondria. Furthermore, 3-deoxyglucosone (3DG), which is generated by the attack of reactive oxygen species on reducing sugars, is also a highly reactive intermediate that, in addition to acting on cells, crosslinks protein molecules together, causing degradation. Reactive oxygen species also have the effect of attenuating the demethylation reaction that reverses DNA methylation, and are involved in the carcinogenesis and aging process by interfering with the post-transplant modification mechanism of genes. From the above, the present invention has the effect of preventing crosslinking of biopolymers, preventing the deterioration of structural molecules, and restoring the cellular environment to a physiological state. As a result, in terms of long-term prognosis, a decrease in the incidence of cancer and a slowdown in the rate of aging that is accelerated in patients with chronic renal failure are expected.

In addition, as emergency treatment for patients with acute renal failure, drug or poison poisoning, severe infections, burns, post-ischemic reperfusion injury, severe pregnancy toxemia, chronic skin ulcers, cerebral hemorrhage and ventricular rupture, and severe hemolytic disease, the blood purification method using the chemical pool drug of the present invention in combination with administration is performed once or several times, and is expected to stop the chain reaction and oxidation reaction of free radicals that increase the fatality rate, thereby contributing to improving the survival rate.

The dialysis device of the present invention, the chemical pool chemicals (acceptor chemicals and donor chemicals), the rinse solution containing the chemical pool chemicals, and the method of using a solution containing the chemical pool chemicals as a rinse solution have been described above based on the illustrated embodiments, but the present invention is not limited thereto. For example, each part constituting the dialysis device can be replaced with any other part having a configuration that can perform the same function. The present invention may also be a combination of any two or more features.

For example, the blood purifier provided in the dialysis device of the present invention is a concept including a hemodialyzer, a hemodialysis filter, a hemofilter, and a plasma separator. Specifically, when the dialysis device is used for hemodialysis, the blood purifier is configured as a hemodialyzer or a dialyzer, as explained based on the above-mentioned embodiment. When the dialysis device is used for hemodialysis filtration, the blood purifier is configured as a hemodialysis filter or a hemodiafilter (a filtration type blood purifier). When the dialysis device is used for hemofiltration, the blood purifier is configured as a hemofilter or a hemofilter (a filtration type blood purifier). When the dialysis device is used for plasma exchange, the blood purifier is configured as a plasma separator or a plasma separator.

Therefore, the "blood purification method" of the present invention, which uses the administration of a chemical pool drug, is a concept that includes hemodialysis, hemodiafiltration, hemofiltration, and plasma exchange. The dialysis device of the present invention is capable of carrying out these blood purification methods. For convenience, the term "chemical pool dialysis" according to the present invention is used as a broad generic term that includes chemical pool hemodiafiltration, chemical pool hemofiltration, and chemical pool plasma exchange.

Hemodiafiltration or hemofiltration uses a large-pore membrane with a large molecular fraction size, making it excellent at removing large molecular compounds. Specifically, by using hemodiafiltration or hemofiltration, the reduction rate of the concentration of large molecular compounds in the blood (e.g. α1-microglobulin (α1-m)) before and after treatment can be increased to about 40-60%.

Furthermore, the use of plasma exchange as a blood purification method can significantly increase the efficiency of removing large molecular compounds (such as α1-m molecules). For example, about half of the α1-m molecules in the blood are bound 1:1 to immunoglobulin A dimers to form large molecules with a molecular weight of 350 kD. For this reason, hemodiafiltration or hemofiltration can only achieve a reduction rate of about 40-60% in the blood concentration of α1-m molecules before and after treatment, but plasma exchange can achieve a reduction rate of about 80%. However, plasma exchange has a significantly higher treatment cost.

Hemodialysis is the most widely used method of blood purification treatment for patients with chronic renal failure, but in recent years the proportion of hemodiafiltration has been increasing. The latter is better than the former at removing large molecular weight compounds in the 20 to 40 kD range.

Therefore, by selecting hemodiafiltration, hemofiltration, or plasma exchange, which are excellent at removing large molecular compounds, instead of hemodialysis, as the blood purification method constituting chemical pool dialysis, it is possible to use large molecular compounds (e.g., α1-m molecules) as the chemical pool drug of the present invention in addition to the small molecular compounds described in the above embodiments. The α1-m molecule (also simply referred to as α1-m) will be described in detail below.

(Chemical reactivity of α1-m)
α1-m is a protein with a strong ability to scavenge reactive oxygen species and free radicals. Specifically, α1-m is a glycoprotein with a molecular weight of 33 kDa, and has physiological functions such as free heme binding ability, scavenging ability through chemical reactions with radicals, and reduction activity.

The chemical reactivity of α1-m is described in detail based on the amino acid sequence of α1-m. In the amino acid sequence of α1-m, the amino acid residues important for radical scavenging and reduction are C34, Y22, K92, K118, H123, K130, and Y132. Here, C is the one-letter abbreviation for the amino acid residues of cysteine, Y is tyrosine, K is lysine, and H is histidine. The number next to the one-letter abbreviation is the residue number.

The amino acid sequence requirement for an α1-m preparation (recombinant α1-m preparation described below) that can be used as a chemical pool drug of the present invention is that it preserves at least three of the following residues: C34, Y22, K92, K118, H123, K130, and Y132, with the original amino acid sequence as the basic structure (main chain). All α1-m that meet this requirement meet the chemical reactivity requirements for a chemical pool drug.

In addition, α1-m also has the ability to bind free heme, which is a harmful substance that generates free radicals and drives the mechanism of oxidative damage. One molecule of α1-m captures two molecules of heme, capturing a maximum of nine radical molecules.

(Stability of α1-m)
α1-m captures harmful substances such as free heme through chemical bonds. Therefore, once α1-m captures free heme, it exists as a stable trap. A stable trap means that there is no possibility of re-release of free radicals or oxidized molecules. In other words, α1-m can exist stably in the body as a free radical trap, so it meets the stability requirements of a chemical pool drug.

(α1-m removal ability)
Furthermore, by selecting hemodiafiltration, hemofiltration or plasma exchange as the blood purification method, it is possible to remove large molecular compounds, so α1-m can also meet the requirements for removal of chemical pool drugs of the present invention.

(Other requirements for α1-m)
The large molecule α1-m meets the requirements of being non-binding, liquid-phase, and non-metabolizable, but is distributed mainly in the extracellular fluid in the body, with less than 10% distributed intracellularly. Although α1-m acts when sprinkled on mitochondria in a test tube, most of the molecules do not reach mitochondria when administered to the body (e.g., by injection). In other words, α1-m does not meet the requirements of accessibility.

In conclusion, α1-m is a substance with strong pharmacological effects that completely meets the three essential requirements for a chemical pool drug. However, when it comes to capturing (or reducing) and removing radicals generated from mitochondria, small molecule compounds are overwhelmingly superior in terms of the size of the chemical equivalent and the ability to reach deep areas (deep and deepest layers) (drug delivery).

Regarding radical capture, let's compare the chemical equivalent per gram of a small molecule with a molecular weight of 330 and α1-m with a molecular weight of 33,000. If you have 1 gram of a small molecule with a molecular weight of 330 and it captures one radical, the equivalent is 3 mEq. On the other hand, if you have 1 gram of α1-m with a molecular weight of 33,000, it can capture nine radicals, so the equivalent is 0.27 mEq. There is a difference of almost 10 times between the two, so the small molecule compound has an overwhelmingly larger capacity at the same dose, which is advantageous. Note that while α1-m is almost 100% stable in the body, it is thought that it is not possible to reduce metabolism to zero with small molecule compounds, so the true difference in effect is less than 10 times.

(Purification of α1-m)
α1-m preparations purified from the urine of healthy humans contain many molecules that have already become radical scavengers and have been degraded, and therefore have poor radical scavenging and reducing abilities and do not have the effects of chemical pool drugs, making them unsuitable.

Therefore, it is preferable to use, as the chemical pool drug of the present invention, a purified product of a recombinant protein in which the human α1-microglobulin gene is introduced into the DNA of another organism and expressed (hereinafter referred to as a "recombinant α1-m preparation"). Recombinant α1-m preparations can be produced, for example, by recombining the human α1-m gene in Chinese hamster ovary cells. Note that the types of cells and animal species used for gene recombination and production are not limited to these.

(Native and mutant recombinant α1-m preparations)
As the chemical pool drug of the present invention, recombinant α1-m preparations of mutant sequences in which base substitutions accompanied by amino acid substitutions have been performed within the range that satisfies the requirements for important residues, as well as recombinant α1-m preparations of native sequences, are suitable. In particular, recombinant α1-m preparations that have been genetically mutated to increase water solubility, recombinant α1-m preparations that have been bound to sugar chains, and recombinant α1-m preparations that do not have the terminal 3-4 residues are preferred.

Specifically, by replacing the hydrophobic residues present on the molecular surface of α1-m with hydrophilic residues, water solubility can be increased, which is useful since it makes it easier to prepare the formulation. Furthermore, binding of glycans to amino acid side chains also contributes to increasing water solubility. For these reasons, recombinant α1-m formulations that have been genetically mutated to increase water solubility and recombinant α1-m formulations to which glycans have been bound are preferred as chemical pool agents of the present invention.

In addition, recombinant α1-m preparations that do not have the C-terminal three residues IPR or four residues LIPR in the amino acid sequence of α1-m have reduced binding to immunoglobulin A, and therefore have enhanced removal properties. For this reason, recombinant α1-m preparations that do not have the terminal three or four residues are preferred as chemical pool agents of the present invention.

The recombinant α1-m preparation is a biological preparation, and as such, its manufacturing costs are very high. From the perspective of medical finances, examples of applications in which administration of recombinant α1-m preparations during chemical pool dialysis in chronic dialysis patients may be appropriate include those with intractable conditions that significantly reduce the quality of life and cannot be improved by the chemical pool dialysis treatment method described in the above embodiment, such as restless leg syndrome, inflammatory joint pain, and limited range of motion due to arthritis.

(Example of administration of recombinant α1-m preparation)
When a recombinant α1-m formulation is used in chemical pool dialysis of a chronic dialysis patient, a replacement fluid containing the recombinant α1-m formulation can be supplied via the replacement fluid supply circuit 400 in the dialysis device 100 described above, but preferably, the recombinant α1-m formulation is administered once or several times after blood purification treatment. For example, in the dialysis device 100, the recombinant α1-m formulation as an injection formulation is administered intravenously at 0.1 g to 2 g per administration after blood purification treatment without using the replacement fluid supply circuit 400. In this case, the dialysis device 100 may be equipped with a drug administration means capable of administering the recombinant α1-m formulation. The dialysis device 100 may also be equipped with a first chemical pool drug supply means (a means for supplying a chemical pool drug via a replacement fluid supply circuit) and a second chemical pool drug supply means (a means for supplying a chemical pool drug without using a replacement fluid supply circuit). Furthermore, the dialysis device 100 is preferably equipped with a control unit capable of independently controlling the first and second chemical pool drug supply means. The improvement effect of administration lasts for several weeks to several months. If inflammatory symptoms recur, chemical pool dialysis using recombinant α1-m preparations is repeated. However, the dosage is not limited to this amount.

In patients who are candidates for acute blood purification treatment, such as those with acute renal failure, drug or poison poisoning, severe infections, burns, post-ischemic reperfusion injury, severe pregnancy toxemia, chronic skin ulcers, cerebral hemorrhage and ventricular rupture, and severe hemolytic disease, the radical scavenging and reducing abilities are reduced or depleted due to the consumption (degradation) of α1-m molecules in the body. Therefore, the administration of recombinant α1-m preparations can reduce the severity of these pathologies involving active oxygen injury. In these patients, renal function is also reduced, so α1-m molecules (nonfunctional degraded protein molecules) that have captured free radicals and changed into scavenger (degraded) are difficult to excrete in the urine and must be removed by blood purification methods.

As soon as possible after the onset of these pathological conditions, 0.1g to 5g of recombinant α1-m preparation is administered intravenously in large amounts. Several hours to several days after administration, blood purification treatments such as hemodialysis filtration, hemofiltration, or plasma exchange are performed to remove non-functional degraded protein molecules and α1-m molecules that have been transformed into them. This constitutes one course, and is carried out once or several times. The dosage per administration is preferably 0.1g to 5g, but is not limited to this.

(Application Example 1)
From here, an application example (for example, solving a problem caused by intravascular hemolysis) using the chemical pool drug of the present invention will be described. In chronic dialysis treatment, a small amount of hemolysis occurs in the circuit (i.e., in the blood vessels) in each extracorporeal circulation treatment, generating free radicals. In chemical pool dialysis, a predetermined total amount of chemical pool drug is administered after each extracorporeal circulation treatment. If the unreacted form of the chemical pool drug administered after the previous extracorporeal circulation treatment remains at the start of the current extracorporeal circulation treatment, even if intravascular hemolysis occurs due to the extracorporeal circulation treatment, the remaining unreacted form of the chemical pool drug (residual chemical pool drug) can suppress the harmful effect of the free radical caused by intravascular hemolysis.

To achieve this effect, it is preferable to increase the amount of the chemical pool drug administered each time over the specified total amount (for example, 0.001 to 0.05 mEq/(hr·kg) when treating a patient with chronic renal failure). This is believed to more reliably prevent the progression of chain reactions of harmful free radicals in the shallow layer (extracellular fluid), deep layer (cytoplasm), and deepest layer (mitochondria) of the chemical pool. From this perspective, it is more preferable to increase the amount of the chemical pool drug administered each time by about 1.1 to 2 times the specified total amount.

Recombinant α1-m preparations are useful for neutralizing radicals and heme in the shallow layer (extracellular fluid) or terminal layer (from the perspective of the radical reaction cascade) of the chemical pool. Therefore, if treatment costs are not taken into consideration, the combined use of small molecule chemical pool drugs and recombinant α1-m preparations is considered to be effective in chronic dialysis patients from the perspective of solving the problems caused by intravascular hemolysis mentioned above.

(Application Example 2)
The combination of small molecule chemical pool drugs and recombinant α1-m is thought to be particularly effective in conditions where intravascular hemolysis is present (e.g., hemolytic uremic syndrome) and conditions where radical production from exogenous causes leads to adverse events (e.g., toxic poisoning). Both of these conditions are covered by acute blood purification therapy. Inflammation, microcirculatory disorders, and tissue destruction are common in these conditions.

As mentioned above, the superiority of α1-m is its stability after radical scavenging, as well as its ability to capture two heme molecules. Heme is formed when hemoglobin leaks from red blood cells during hemolysis and separates from the globin through decomposition. Heme itself is not a radical, but it is a very harmful molecule that promotes radical production. Fetal hemoglobin F and free heme damage vascular endothelial cells throughout the mother's body, resulting in a disease called eclampsia, a severe form of preeclampsia. Therefore, α1-m is effective in neutralizing radicals in blood and interstitial fluid, and heme that produces radicals.

On the other hand, small molecule chemical pool drugs cannot capture heme to the same extent or greater than themselves, but they can act to neutralize radicals generated by heme. For these reasons, it is believed that the combined administration of small molecule chemical pool drugs and recombinant α1-m preparations is particularly effective for the pathologies covered by the above-mentioned acute blood purification treatments.

(Application Example 3)
Acute blood purification is usually performed only a few times, so expensive treatment is allowed, but chemical pool dialysis using recombinant α1-m preparations in chronic dialysis patients may be limited to some patients with intractable pathologies due to rising medical costs. Therefore, for other patients, it is desirable to practice spontaneous pseudo-chemical pool dialysis caused by endogenous α1-m production. In other words, it is possible to rely on pseudo-chemical pool dialysis, which removes α1-m from the body and promotes new production in the liver. Below, we will explain physiological production and metabolism of α1-m, and then explain pseudo-chemical pool dialysis.

α1-m in the body is produced in the liver. Because of its high renal metabolic excretion rate, its blood half-life is only a few hours, but its production rate in the liver is also high, so that its blood concentration is always kept in balance at 10-20 mg/L. When renal function is normal, α1-m traps that have captured free radicals and other substances are quickly filtered by the glomeruli of the kidney and excreted in the urine either as is or in the form of decomposed fragments after being reabsorbed by the renal tubules. In this way, some of the free radicals produced in the body are excreted from the body.

On the other hand, in patients with renal failure, this metabolic excretion function is lacking, so α1-m that has become a free radical trap remains in the body. Specifically, in patients undergoing chronic dialysis, the blood concentration rises due to the lack of renal metabolic excretion. When the blood concentration rises, production in the liver is suppressed by feedback control and falls. For this reason, the blood concentration does not rise indefinitely, and the balance is ultimately maintained at a concentration of 120 to 150 mg/L. In patients undergoing chronic dialysis, there is almost no turnover (new replacement) of α1-m molecules. Therefore, even at a high concentration of 120 to 150 mg/L, the radical trapping and reducing abilities of α1-m are already exhausted. In other words, the α1-m in the body of a chronic dialysis patient has become a non-functional degraded protein molecule. In normal hemodialysis, almost no α1-m with a molecular weight of 33 kD is removed, so this level is maintained and no significant fluctuations occur.

In such chronic dialysis patients, if hemodiafiltration or hemofiltration, which are better at removing larger molecules, are used instead of hemodialysis, approximately 200-300 mg of α1-m can be removed in a single 4-5 hour treatment. The blood concentration can also be reduced by approximately 40-60%. However, the blood concentration continues to rise over the 2-3 days until the next treatment, eventually returning to the original blood concentration of 120-150 mg/L. This means that during the same period, the 200 mg of α1-m molecules removed by the previous blood purification method were newly synthesized in the liver to offset the decrease. Since the new α1-m has radical scavenging and reducing abilities, it is thought to exert an effect only in proportion to its chemical equivalent. If the chemical equivalent is the same, the endogenous α1-m produced by the liver and the recombinant α1-m preparation administered from outside have the same effect. In blood purification therapy, by actively removing α1-m, which has become a non-functional, degraded protein molecule, endogenous production of α1-m is induced from the liver, essentially creating a spontaneous pseudo-chemical pool dialysis.

In this way, by practicing hemodiafiltration or hemofiltration, it is possible to rotate the molecule of α1-m, a protein that has a strong ability to scavenge reactive oxygen species and free radicals. This is effective in improving oxidative stress defense and improving the state of cell proliferation inhibition in renal failure.

(Application Example 4)
Chemical pool dialysis can be applied to the treatment of paraquat poisoning in the acute blood purification field. First, paraquat poisoning will be explained. The chemical name of paraquat is 1,1'-dimethyl-4,4'-bipyridinium dichloride. Paraquat is still used today as a pesticide, and if accidentally ingested, it is absorbed into the body from the intestinal tract, enters the cells, and reaches the outer mitochondrial membrane. Once it reaches the outer mitochondrial membrane, paraquat undergoes one-electron reduction to become a paraquat radical, which induces an intense systemic radical chain reaction. As a result, in addition to a large amount of active oxygen damage, the cells die due to the depletion of NADPH. Clinical symptoms begin with vomiting immediately after ingestion, and over time, diarrhea, abdominal pain, sores in the mouth, and ulcer formation occur. In severe cases, kidney dysfunction, liver dysfunction, and shock progress, and interstitial pneumonia and pulmonary fibrosis also occur, leading to death. The lethal dose of paraquat is several tens of mmol.

Because tissue damage caused by the generation of active oxygen is at the core of the pathology of paraquat poisoning, attempts have been made to administer large amounts of the antioxidant α-tocopherol (vitamin E), but in severe cases where organ damage has developed, it is virtually impossible to save the patient's life. This is thought to be related to the fact that vitamin E does not sufficiently block the radical chain reaction, that vitamin E is fat-soluble and dissolves in membranes, degrading membrane lipids, and that vitamin E radicals after chemical reactions are not excreted from the body.

Considering the above relationship, chemical pool dialysis can be applied to the treatment of paraquat poisoning. Specifically, the protocol for chemical pool dialysis for paraquat poisoning involves intravenous administration of 10 to 100 mmol of reduced ascorbic acid (vitamin C) and reduced glutathione as chemical pool drugs as soon as possible after exposure. However, chemical pool drugs are not limited to these two drugs. The chemical pool drugs are administered intravenously prior to or in parallel with gastrointestinal irrigation. Then, within one day (preferably within four hours), blood purification treatment is performed for 4 to 6 hours to remove the chemical pool drugs. A second administration of chemical pool drugs is performed at the end of blood purification. Then, a second blood purification therapy is performed within one day. Thereafter, the administration of chemical pool drugs and blood purification are repeated for several courses. When continuous blood purification treatment is performed instead of intermittent treatment as blood purification treatment, additional administration of chemical pool drugs is performed every 4 to 24 hours during the continuous blood purification treatment to replenish the reduced concentration of the remaining chemical pool drugs.

In chemical pool dialysis for paraquat poisoning, the radical chain reaction can be blocked by administering 0.5 to 10 g of recombinant α1-m alone as a chemical pool drug. However, as a blood purification method to remove chemical pool drugs, hemodialysis filtration or hemofiltration is used, which is relatively more efficient at removing large molecules than hemodialysis, and a large-pore membrane is selected as a blood purifier. Furthermore, adding plasma exchange while replenishing plasma drugs can remove α1-m molecules more efficiently. Although a certain degree of effect can be expected even when recombinant α1-m is used alone as a chemical pool drug, it is preferable to use it in combination with a chemical pool drug of a small molecular compound. Examples of chemical pool drugs of small molecular compounds to be used in combination include reduced ascorbic acid (vitamin C) and reduced glutathione, but are not limited to these two drugs. The treatment schedule, which involves administering chemical pool drugs and repeating several courses of blood purification, is the same as the above protocol.

(Application Example 5: Treatment of side effects of anticancer drugs)
Another application of chemical pool dialysis in the field of acute blood purification is the treatment of severe cytotoxic side effects after the administration of anticancer drugs. Anticancer antibiotics of the anthraquinone (anthracycline) family, such as adriamycin, have a quinone structure in their molecules, and after being taken up into cells, they undergo one-electron reduction in microsomes to produce semiquinone radicals, which induce radical chain reactions. Cardiotoxicity is well known as a severe overt side effect. When heart failure develops, the condition deteriorates irreversibly and often becomes fatal. When symptoms of cardiotoxicity appear, administration of chemical pool drugs is started as soon as possible. The protocol for this treatment is similar to that for the treatment of paraquat poisoning.

The following examples are specifically described, but the present invention is not limited to the following examples as long as they do not exceed the gist of the invention.

Example 1
Chemical pool dialysis was performed on a chronic dialysis patient weighing 60 kg who was receiving a normal blood purification treatment (blood flow of 200-300 mL/min, 4-5 hours of hemodialysis) performed on a schedule of three times a week. Specifically, 200 mL of aqueous solution (rinse solution) containing 2-(α-D-glucopyranosyl) (methyl)methyl-2,5,7,8-tetramethylchroman-6-ol (FIG. 9), an acceptor drug that captures free radicals, at a chemical equivalent of 10 mEq was prepared as the chemical pool drug of the present invention. This rinse solution was placed in a dialysis device equipped with a high-performance membrane (dialysis membrane with a clearance value of 150 mL/min). At the end of the blood purification treatment, this rinse solution was used for blood recovery in the extracorporeal blood circuit of the dialysis device, and the entire amount was administered to the patient. The administered chemical pool drug was removed from the body by the next blood purification treatment after a treatment intermission period (43-44 hours). This cycle was repeated at the end of each blood purification treatment.

Example 2
Chemical pool dialysis was performed on a chronic dialysis patient weighing 60 kg who was receiving a normal blood purification treatment similar to that in Example 1. Specifically, 300 mL of an aqueous solution (rinse solution) containing 200 mEq of 2-(α-D-glucopyranosyl) (isopropyl)methyl-2,5,7,8-tetramethylchroman-6-ol (FIG. 10), an acceptor drug that captures free radicals, was prepared as the chemical pool drug of the present invention. This rinse solution was placed in a dialysis device equipped with a high-performance membrane (dialysis membrane with a clearance value of 200 mL/min). At the end of blood purification treatment, this rinse solution was used for blood recovery in the extracorporeal blood circuit of the dialysis device, and the entire amount was administered to the patient. The administered chemical pool drug was removed from the body by the next blood purification treatment after a treatment intermission period (43 to 44 hours). This cycle was repeated every time blood purification treatment was completed.

Example 3
Chemical pool dialysis was performed on a chronic dialysis patient weighing 60 kg who was receiving a normal blood purification treatment similar to that in Example 1. Specifically, 500 mL of an aqueous solution (rinse solution) containing 100 mEq of D-lysyl-tyrosine (a dipeptide consisting of two D-amino acid residues) having a non-physiological optical isomer structure, which is an acceptor drug that captures free radicals, was prepared as the chemical pool drug of the present invention. This rinse solution was placed in a dialysis device equipped with a high-performance membrane (dialysis membrane with a clearance value of 150 mL/min). At the end of blood purification treatment, this rinse solution was used for blood recovery in the extracorporeal blood circuit of the dialysis device, and the entire amount was administered to the patient. The administered chemical pool drug was removed from the body by the next blood purification treatment after a treatment intermission period (43 to 44 hours). This cycle was repeated every time blood purification treatment was completed. Note that D-peptide radicals that have captured radicals and D-peptides that have been oxidized are not easily decomposed in the body and remain stable during the treatment intermission period, so they can be removed by the next dialysis treatment.

Example 4
Chemical pool dialysis was performed on a chronic dialysis patient weighing 60 kg who was receiving the same normal blood purification treatment as in Example 1. Specifically, 500 mL of an aqueous solution containing D-lysyl-tyrosyl-cysteine (a tripeptide consisting of three D-amino acid residues) having a non-physiological optical isomer structure, which is an acceptor drug that captures free radicals, at a chemical equivalent of 100 mEq was prepared as the chemical pool drug of the present invention. A total of 15 g of protein-constituting L-amino acids was added to this aqueous solution to prepare a rinse solution containing L-amino acids and D-lysyl-tyrosyl-cysteine. This rinse solution was placed in a dialysis device equipped with a high-performance membrane (dialysis membrane with a clearance value of 150 mL/min). At the end of blood purification treatment, this rinse solution was used for blood recovery in the extracorporeal blood circuit of the dialysis device, and the entire amount was administered to the patient. Since the osmotic pressure ratio of the rinse solution to physiological saline was 3 to 4, slow administration (recovery operation) was performed over several minutes or more. The administered chemical pool drug was removed from the body by the next blood purification treatment after a treatment intermission period (43-44 hours). This cycle was repeated every time blood purification treatment was completed.

Example 5
Chemical pool dialysis was performed on a chronic dialysis patient weighing 60 kg who was receiving a normal blood purification treatment similar to that in Example 1. Specifically, 300 mL of an aqueous solution (rinse solution) containing reduced ascorbic acid and glutathione or glutathione alkyl ester (a donor drug that is an H donor) in a total chemical equivalent of 80 mEq was prepared as the chemical pool drug of the present invention. The content ratio of reduced ascorbic acid to glutathione or glutathione alkyl ester was 1:3 in terms of chemical equivalent ratio. This rinse solution was placed in a dialysis device equipped with a high-performance membrane (dialysis membrane with a clearance value of 150 mL/min). At the end of blood purification treatment, this rinse solution was used for blood recovery in the extracorporeal blood circuit of the dialysis device, and the entire amount was administered to the patient. The administered chemical pool drug was removed from the body by the next blood purification treatment after a treatment intermission period (43 to 44 hours). This cycle was repeated every time blood purification treatment was completed.
In addition, the function of glutathione alkyl esters is to serve as substrates for glutathione peroxidase and exert their reducing properties, similar to glutathione; however, in terms of transport, they have the advantage of having superior membrane permeability to glutathione, making them easier to reach mitochondria.

Example 6
Chemical pool dialysis was performed on a burn patient weighing 60 kg who required acute blood purification treatment. Specifically, as the chemical pool drug of the present invention, 500 mL of aqueous solution containing 2-(α-D-glucopyranosyl) (methyl)methyl-2,5,7,8-tetramethylchroman-6-ol, an acceptor drug that captures free radicals, and glutathione or glutathione alkyl ester (H donor) in a total chemical equivalent of 120 mEq was prepared. The content ratio of 2-(α-D-glucopyranosyl) (methyl)methyl-2,5,7,8-tetramethylchroman-6-ol to glutathione or glutathione alkyl ester was 2:1 in chemical equivalent ratio. This aqueous solution was administered intravenously to the patient using a dialysis device as soon as possible after the occurrence of an adverse event. Then, after the detoxification reactions such as radical scavenging and reduction reactions reached equilibrium (usually after several hours to several days), highly efficient blood purification treatment was performed to remove the reaction products. For this purpose, acute blood purification treatment must have a better efficiency of removing small molecular substances than normal chronic blood purification treatment of the slow and continuous type. For this reason, the blood purifier of the dialysis device uses a dialysis membrane with a clearance value of 200 ml/min, and blood purification treatment is performed for 4 hours or more. When the clearance value is 100 ml/min, blood purification treatment is preferably performed for 6 hours or more, and when the clearance value is 150 ml/min, blood purification treatment is preferably performed for 5 hours or more. Administration of a chemical pool drug and the subsequent blood purification treatment constitute one course, and three courses are repeated once every three days. When continuous treatment is performed, additional chemical pool drug is administered during blood purification treatment when about 10 to 50% of the chemical equivalent of the chemical pool drug administered into the body is removed immediately after administration.

The chemical pool dialysis performed in Examples 1 to 5 was able to efficiently prevent the progression of the chain reaction of highly chemically reactive and harmful free radicals that are constantly generated in the metabolic process. Such chemical pool dialysis is expected to reduce the rate of progression of comorbid pathologies such as muscle atrophy, motor dysfunction, arteriosclerosis, cancer, and accelerated aging, which are caused by the vicious cycle of reduced ability to eliminate reactive oxygen species in chronic renal failure and mitochondrial disorder, and contribute to improving the survival and quality of life of patients, as well as the preventive medical benefit of reducing medical expenses spent on treating complications on a national level. The reduced ascorbic acid that constitutes the chemical pool drug used in Example 5 does not reach mitochondria, but is reduced by glutathione or glutathione alkyl esters after capturing free radicals that have leaked out of mitochondria. Furthermore, glutathione or glutathione alkyl esters reach mitochondria. It is therefore presumed that the chain reaction of free radicals can be prevented at multiple stages.

In addition, by applying chemical pool dialysis as in Example 6 to the fields of acute blood purification treatment and emergency treatment, it is expected that the survival rate of patients with acute renal failure, drug overdose or poisoning, severe infections, burns, post-ischemic reperfusion injury, severe preeclampsia, chronic skin ulcers, cerebral hemorrhage and ventricular rupture, and severe hemolytic disease can be improved.

1. Blood purification treatment 2. Administration of chemical pool drugs 3. Profile of the amount (mEq) of drugs distributed in the extracellular fluid in the chemical pool 4. Profile of the amount (mEq) of drugs distributed in the cytoplasm in the chemical pool 5. Profile of the amount (mEq) of drugs distributed in the mitochondria in the chemical pool 6. Change in size (mEq) of the chemical pool when the intrinsic clearance value is 1 mL/min 7. Change in size (mEq) of the chemical pool when the intrinsic clearance value is 2 mL/min 8. Change in size (mEq) of the chemical pool when the intrinsic clearance value is 4 mL/min 9. Change in size (mEq) of the chemical pool when the intrinsic clearance value is 8 mL/min 10. Change in size (mEq) of the chemical pool when the intrinsic clearance value is 12 mL/min 11. Change in the amount (mEq) of drugs present in the mitochondria when the intrinsic clearance value is 1 mL/min 12 Change in the amount of drug present in mitochondria (mEq) when the intrinsic clearance value is 2 mL/min 13 Change in the amount of drug present in mitochondria (mEq) when the intrinsic clearance value is 4 mL/min 14 Change in the amount of drug present in mitochondria (mEq) when the intrinsic clearance value is 8 mL/min 15 Change in the amount of drug present in mitochondria (mEq) when the intrinsic clearance value is 12 mL/min 16 Change in chemical pool size (mEq) when the blood purification clearance value is 100 mL/min 17 Change in chemical pool size (mEq) when the blood purification clearance value is 150 mL/min 18 Change in chemical pool size (mEq) when the blood purification clearance value is 200 mL/min 19 Change in chemical pool size (mEq) when the blood purification clearance value is 250 mL/min 20 Change in chemical pool size (mEq) when the blood purification clearance value is 300 mL/min
C1: Electromagnetic opening and closing clamp
C2: Electromagnetic opening and closing clamp
D: Blood purifier
D1: Thin tube
P1: Dialysis fluid pump (constant flow rate system)
P2: Dialysis fluid drainage pump (constant flow rate system)
P3: A third dialysis fluid pump that rotates in both directions
P4: A blood pump that rotates in both directions
R: Rinse solution containing chemical pool agents
s: Connection point between the patient's blood vessel on the blood drainage side and the blood circuit
t: Confluence of fluid supply circuit and blood circuit
u: Connection point between the arterial side of the blood circuit and the blood purifier D
v: Connection point between the patient's blood vessel on the blood return side and the blood circuit 100: Dialysis device 200: Dialysis fluid circuit 300: Extracorporeal blood circuit (blood circuit)
310: Arterial blood circuit 320: Venous blood circuit 400: Replacement fluid supply circuit

Claims (10)

An acceptor molecule capable of capturing free radicals generated in the body of a patient receiving blood purification treatment using a blood purifier, or
A chemical pool drug comprising at least one donor molecule capable of inhibiting an oxidation reaction by donating an electron as a hydrogen atom and/or a single electron to an oxidized biomolecule subjected to an oxidation reaction in the body of the patient to whom the blood purification treatment by the blood purifier is administered,
The at least one of the acceptor molecule or the donor molecule is at least one of a small molecule compound having a molecular weight of 1000 D or less or a large molecule compound having a molecular weight of 20 kD or more;
The small molecule compound comprises at least one of reduced ascorbic acid or reduced glutathione;
The large molecule compound comprises recombinant α1-microglobulin,
The chemical pool drug is administered to the patient receiving the blood purification treatment by the blood purifier at a chemical equivalent of 0.001 mEq/(hr kg) or more, based on the length (hr) of the intertreatment period in the blood purification treatment and the body weight (kg) of the patient receiving the blood purification treatment, and is then removed from the patient's body by the blood purifier. 2. The chemical pool drug according to claim 1 , wherein the total amount of said chemical equivalents per one blood purification treatment is 0.001 to 20 mEq/(hr·kg). 3. The chemical pool agent of claim 2 administered in greater than said total amount of said chemical equivalents. 4. The chemical pool agent according to claim 1, wherein the acceptor molecule after capturing the free radical and the donor molecule after donating the electron remain stable for a period of at least 3 hours. The chemical pool agent of claim 1 , wherein the small molecule compound comprises a peptide consisting of 2 or 3 D-amino acid residues.
The chemical pool agent of claim 1 , wherein the recombinant α1-microglobulin comprises at least one of an original recombinant α1-microglobulin or a mutant recombinant α1-microglobulin . An injection preparation comprising the chemical pool agent according to any one of claims 1 to 6 ,
The chemical pool drug is administered to a patient undergoing blood purification treatment using a blood purifier at a chemical equivalent of 0.001 mEq/(hr kg) or more, based on the length of the intertreatment period (hr) in the blood purification treatment and the body weight (kg) of the patient undergoing the blood purification treatment, and is then removed from the patient's body by the blood purifier. An oral preparation comprising the chemical pool agent according to any one of claims 1 to 6 ,
The chemical pool drug is administered to a patient undergoing blood purification treatment using a blood purifier at a chemical equivalent of 0.001 mEq/(hr kg) or more, based on the length of the intertreatment period (hr) in the blood purification treatment and the body weight (kg) of the patient undergoing the blood purification treatment, and is then removed from the patient's body by the blood purifier. An infusion preparation comprising the chemical pool drug according to any one of claims 1 to 6 ,
An infusion preparation characterized in that the chemical pool drug is administered to a patient undergoing blood purification treatment using a blood purifier at a chemical equivalent of 0.001 mEq/(hr kg) or more, based on the length (hr) of the intertreatment period in the blood purification treatment and the body weight (kg) of the patient undergoing the blood purification treatment, and is then removed from the patient's body by the blood purifier. A dialysis apparatus comprising: a blood purifier; an extracorporeal blood circuit connected to the blood purifier for circulating blood; a supply means connected to the extracorporeal blood circuit and including the infusion preparation according to claim 9 ; and a dialysate circuit for supplying a dialysate to the blood purifier,
The dialysis apparatus according to claim 1, wherein the supply means is configured to supply the infusion preparation to the extracorporeal blood circuit.