JP2009114261A - Uremia therapeutic drug and uremia therapeutic food - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a uremia therapeutic drug and uremia therapeutic food. <P>SOLUTION: In the uremia therapeutic drug, a metal salt of a crosslinked cellulose derivative represented by the following general formula (I) is an active ingredient. In the formula R-O-A (I), R represents a crosslinked cellulose residue, A represents a functional group (a) having cation exchangeability. The functional group having cation exchangeability is a sulfonic acid group, a carboxyl group, a phosphoric acid group, and the like. A concrete compound for the crosslinked cellulose derivative includes a sulfated cellulose calcium salt and the like. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to a uremic remedy and a food for uremic amelioration.

  Decreased renal function causes uremia, and the manifestation of uremia due to the progression of renal failure is life-threatening.

  Currently, hemodialysis, which is the mainstream for the treatment of renal failure, is time-constrained, and the patient undergoing hemodialysis is subject to great time constraints, physical, mental and economic burdens.

  Accordingly, an object of the present invention is to provide a uremic remedy.

  As a sodium absorption inhibitor, a potassium absorption inhibitor and a phosphorus absorption inhibitor, the present inventors inhibit the absorption of sodium, potassium or phosphorus overdose in the gastrointestinal tract, thereby allowing sodium, potassium or phosphorus to be absorbed. A metal salt of a crosslinked cellulose derivative that has excellent action to promote excretion into feces and is lightly digestive tract disorder has been developed (Japanese Patent Application No. 2006-349270).

  Subsequent studies by the present inventors have found that a metal salt of a crosslinked cellulose derivative has an excellent life prolonging effect on nephrectomized rats. This is presumed that the causative substance of uremia secreted from the digestive tract or produced in the intestine is trapped by the metal salt of the crosslinked cellulose derivative.

  Based on the above findings, the present inventors have completed the present invention. That is, the present invention provides the following uremic remedies and foods.

  [1] A uremia-improving drug comprising a metal salt of a crosslinked cellulose derivative represented by the following general formula (I) as an active ingredient.

R-O-A (I)
{In Formula (I), R represents a crosslinked cellulose residue, and A represents a functional group a having a cation exchange ability. }
[2] The uremia-improving drug according to [1], wherein the degree of substitution of the hydroxyl group of the glucose unit of the crosslinked cellulose derivative with the functional group a is 1.2 or more.

  [3] The uremia-improving drug according to [1], wherein the degree of substitution of the hydroxyl group of the glucose unit of the crosslinked cellulose derivative with the functional group a is 1.4 or more.

  [4] The uremia-improving drug according to [1], wherein the degree of substitution of the hydroxyl group of the glucose unit of the crosslinked cellulose derivative with the functional group a is 1.5 or more.

  [5] The uremia-improving drug according to any one of [1] to [4], wherein the functional group a is selected from the groups represented by the following general formulas (II) to (V): .

{In Formulas (II) to (V), alk represents an alkylene group having 1 to 6 carbon atoms, l represents an integer of 0 to 5, m represents 0 or 1, and n represents an integer of 0 to 2. To express. }
[6] The functional group a is represented by the following general formulas (II-1), (II-2), (III-1) to (III-5), (IV-1), (V-1) and (V- The uremic ameliorating drug according to [5] above, which is selected from 2).

  [7] The uremia-improving drug according to [6], wherein the combination of the plural kinds of functional groups a is any one of the following (c-1) to (c-3).

  [8] The uremia-improving drug according to any one of [1] to [7], wherein the crosslinked cellulose derivative is produced using crystalline cellulose.

  [9] The uremia-improving drug according to any one of [1] to [8], wherein the crosslinked cellulose derivative is produced using epichlorohydrin as a crosslinking agent.

  [11] A uremia-improving food, comprising the metal salt of the cellulose derivative according to any one of [1] to [9].

  The crosslinked cellulose derivative can be produced by a production method including a crosslinking step and a substituent introduction step. The substituent introduction step is preferably performed after the crosslinking step from the viewpoint of the stability of the substituent. That is, a cross-linking step of cross-linking cellulose with a cross-linking agent to obtain cross-linked cellulose, and a hydrogen atom in a part or all of the remaining hydroxyl groups not used for cross-linking of the cross-linked cellulose with a functional group a having a cation exchange ability It is preferably obtained by a substituent introduction step for substitution.

  As the cellulose, various known celluloses can be used, and the molecular weight thereof is not particularly limited, however, crystalline cellulose (published by the Japanese Pharmacopoeia) having a uniform degree of polymerization and a certain range is preferable.

  The crosslinking step can be carried out by reacting cellulose with a crosslinking agent. Cross-linking agents include glyoxal, dialdehyde starch, aldehydes such as polyacrolein, methylol compounds such as N-methylol melamine and trimethylol melamine, active vinyl compounds such as divinyl sulfone, epichlorohydrin, epibromohydrin, ethylene Glycol diglycidyl ether, propylene glycol diglycidyl ether, glycerol polyglycidyl ether, pentaerythritol polyglycidyl ether, neopentyl glycol diglycidyl ether, 1,4-butanediol diglycidyl ether, 1,4-bis (2,3-epoxy Propoxy) butane, 1,4-bisglycidoxybutane, 1,2-bis (2,3-epoxypropoxy) ethylene, 1,-(2,3-epoxypropyl) -2,3 Epoxy compounds such as epoxycyclohexane, tartaric acid, citric acid, malic acid, maleic acid, fumaric acid, succinic acid, adipic acid, sebacic acid, aspartic acid, glutaric acid, tricarballylic acid, butanetetracarboxylic acid, polymaleic acid, polyacrylic Acid, polycarboxylic acid such as polyacrylic acid-maleic acid copolymer, diisocyanate compound, oxiranylmethanol, N-ethylbis (2-chloroethyl) amine, methylvinyldiacetoxysilane, dimethyldiacetoxysilane, triglycidyltris ( 2-hydroxyethyl) isocyanurate, formaldehyde, glutaraldehyde, crotonaldehyde, 4,5-dihydroxyethylene urea, divinyl sulfone, preferably halohydrins, glycidyl ether Class, an epoxy compound such as epoxy alkanes, particularly preferably epichlorohydrin.

  Two or more kinds of these crosslinking agents can be used.

  The usage-amount of the said crosslinking agent is 0.1-500 weight part with respect to 100 weight part of cellulose, Preferably, it is 100-300 weight part. More preferably, it is 150-200 weight part. Within these ranges, it is useful because sufficient crosslinking is obtained and no unreacted product remains.

  For the purpose of smoothly proceeding with the crosslinking reaction by the crosslinking agent, a crosslinking reaction catalyst can be appropriately used depending on the type of the crosslinking agent. Examples of the crosslinking reaction catalyst include phosphoric acid, sodium hypophosphite, potassium dihydrogen phosphate, sodium phosphate and other inorganic compounds such as sulfuric acid and hydrochloric acid when the crosslinking agent is a polycarboxylic acid. Sodium carbonate, sodium acetate, titanium compounds and the like, and when an epoxy compound is used as a crosslinking agent, basic compounds such as sodium hydroxide and potassium hydroxide, primary amines, secondary amines, 3 Examples include amines such as quaternary amines, quaternary ammonium salts, imidazole compounds, alcohols and water.

  The usage-amount of the said crosslinking reaction catalyst is 1-200 weight part with respect to 100 weight part of said crosslinking agents, Preferably, it is 80-150 weight part.

  A method for crosslinking the cellulose with the crosslinking agent is not particularly limited, and examples thereof include a method described in JP-A-63-54160. Specifically, there are many methods such as a method by dry crosslinking in which cellulose is impregnated with an aqueous crosslinking agent solution, dehydrated and dried at high temperature, and an aqueous solution crosslinking method in which the cellulose is crosslinked in an aqueous crosslinking agent solution. A two-layer cross-linking method in which a cross-linking agent and cellulose are vigorously stirred in a cross-linking catalyst aqueous solution in a two-layer system to cross-link is more preferable.

  Crosslinking of the cross-linked cellulose may be at any position, and may be cross-linking in one cellulose or cross-linking of celluloses.

  The degree of crosslinking of the metal salt of the crosslinked cellulose derivative is preferably 0.01 or more.

  The degree of crosslinking can be calculated as follows.

  Here, a case where cellulose used for the reaction has a molecular weight of 39000 (average polymerization degree of 240) and epichlorohydrin is used as a crosslinking agent will be described as an example.

  Epichlorohydrin increases the cross-linked structure portion of the sugar chain of cellulose as shown in the following structural formula. The molecular weight of this increasing part is 56.

When the charged amount of cellulose before cross-linking and the weight of cross-linked cellulose after cross-linking is a value obtained by subtracting the moisture content (analytical value) and increased by 4%, the molecular weight of cellulose is 39000.
39000 × 0.04 = 1560,
Since the molecular weight of the increasing molecular structure is 56,
1560 ÷ 56 ≒ 28
From the calculation, the crosslinked structure portion is calculated to be about 28 mol. That is, the ratio of crosslinking / cellulose is 28/1.

When this is converted per sugar hydroxyl group,
Since 1 mol of cellulose has 240 glucose (average polymerization degree) and 3 hydroxyl groups per glucose unit, the number of hydroxyl groups per 1 mol of cellulose is:
240 × 3 = 720
And the ratio to the crosslinked structure portion is 49/1 as described above,
49/720 = 0.068
Is calculated. That is, 0.068 per hydroxyl group, and 0.068 can be defined as the “crosslinking degree” of the crosslinked cellulose of the above example.

The number of moles of cellulose was converted using 1 mole of glucose unit = 162 g. That is, since the glucose unit in 1 mol of cellulose has an average molecular weight of 39000,
162 ÷ 39000 ≒ 4.15mmol
Is converted.

  The degree of crosslinking is preferably 0.01 or more. More preferably, it is 0.01 or more and 0.30 or less, and further preferably 0.030 or more and 0.20 or less.

  The degree of crosslinking can be controlled by the ratio of the dose of the base or acid for activating the functional group to cellulose and the dose of the crosslinking agent. In addition, since it is a solid-liquid reaction, it can be controlled depending on the type of solvent used and also when the number of solvents is two or more. In order to control the degree of cross-linking within a preferable range, it is preferable that the ratio of the dose of the base or acid for activating the functional group to cellulose and the dose of the cross-linking agent is mainly used.

  For example, when the cross-linking agent is epichlorohydrin, the hydroxyl group is a functional group, so sodium hydroxide is used as the base. However, when the ratio of epichlorohydrin to sodium hydroxide is constant, the epithelium is epithelial. The degree of crosslinking increases as the amount ratio of chlorohydrin is increased. However, when sodium hydroxide is excessively large relative to epichlorohydrin, the decomposition reaction of cellulose may proceed in parallel, and does not necessarily lead to an improvement in the degree of crosslinking.

  In addition, when a large amount of a polar solvent such as methanol or isopropyl alcohol is used as the solvent to be used, decomposition (lower molecular weight) of cellulose proceeds and the degree of crosslinking may increase, but the recovery rate may decrease. is there.

  Epichlorohydrin / sodium hydroxide is preferably 1/2 to 1/4, more preferably 1 / 2.3 to 1/3. In terms of the molar ratio of epichlorohydrin and sodium hydroxide. 2. Furthermore, when expressed in a quantitative ratio with cellulose, cellulose / sodium hydroxide / epichlorohydrin is 1/6/3 to 1/20/5, more preferably 1/7/3 to 1/16/5. .

  The solvent used is a three-phase system such as n-heptane / methanol / water, n-heptane / isopropyl alcohol / water, or a two-phase system such as n-heptane / water. A nonpolar solvent and a polar solvent (water N-heptane / water is more preferred.

  Furthermore, by dividing the crosslinking agent, that is, by dividing the crosslinking agent and increasing the number of administrations, the amount of the crosslinking agent that is hydrolyzed without being used in the crosslinking reaction can be suppressed, and the degree of crosslinking can be further increased. Further, for example, the degree of crosslinking can be further increased by increasing the stirring efficiency by increasing the stirring speed, changing the shape of the stirring blade, adding a baffle, or reducing the reaction scale.

  The metal salt of a crosslinked cellulose derivative is a metal salt in which crosslinking and functional groups are introduced into cellulose. The metal salt of the crosslinked cellulose derivative is represented by the following general formula (I).

  R-O-A (I)

  {In Formula (I), R represents a crosslinked cellulose residue, and A represents a functional group a having a cation exchange ability. }

  Examples of the functional group a having cation exchange ability include groups having a carboxyl group, a sulfonic acid group, a phosphonic acid group, a phosphoric acid group, and the like, and preferably represented by the following general formulas (II) to (V). Group.

  {In Formulas (II) to (V), alk represents an alkylene group having 1 to 6 carbon atoms, l represents an integer of 0 to 5, m represents 0 or 1, and n represents an integer of 0 to 2. To express. }

  The crosslinked cellulose derivative is a cellulose ether substituted with a functional group a having a cation exchange ability, and the functional group a having a cation exchange ability substituted for the cellulose derivative may be one kind or two or more kinds.

  Preferred examples of the functional group a include the following.

  Moreover, the crosslinked cellulose derivative may have a functional group a having two or more kinds of cation exchange ability, for example, different types of cations composed of combinations shown in the following (c-1) to (c-3). A crosslinked cellulose derivative constituted by a functional group having exchange ability is preferred.

  Further, the group having a phosphonic acid group or a phosphoric acid group represented by the above formula (IV) or (V) is sufficient if at least one hydroxyl group is present in the functional group. The hydroxyl group in the acid group may be substituted with an alkoxy group, a phosphonic acid group, a thiol group or the like as necessary. Specifically, the following groups are also included in the functional group a having a cation exchange ability.

  The functional group introduction step is preferably performed after the crosslinking step.

  For example, when the functional group a is a skeleton of the general formula (III), as a sulfating agent for introducing a sulfate group into a hydroxyl group of cellulose, concentrated sulfuric acid, fuming sulfuric acid, sulfuric anhydride, sulfuric anhydride / DMF complex, sulfuric anhydride / Examples thereof include a pyridine complex, an anhydrous sulfuric acid / triethylamine complex, and chlorosulfonic acid, and an anhydrous sulfuric acid / DMF complex is preferably used.

  The sulfated crosslinked cellulose can be converted directly into a metal salt after removing the unreacted crosslinking agent and its derivative together with a solvent, for example, with calcium chloride added in advance, or an alcohol and an inorganic salt such as methanol or isopropanol. And then vigorously stirring to obtain a metal salt. Moreover, after manufacturing a calcium salt once, a metal salt can also be obtained by substitution reaction with an inorganic salt.

  As the metal forming the metal salt, alkali metals such as potassium, alkaline earth metals such as calcium and magnesium, and metals such as iron can be used. Among these, potassium, calcium, magnesium, and iron are preferable, and calcium is most preferable from the viewpoint of ion exchange efficiency and the point that it is allowed to be released into the blood or the body.

  The metal salt of the crosslinked cellulose derivative can suppress the damage to the digestive tract, which is considered to be due to crosslinking.

  The degree of substitution of the metal salt of the crosslinked cellulose derivative represented by the general formula (I) is preferably 1.2 or more, more preferably 1.4 or more, and further preferably 1.5 or more.

  The degree of substitution can be calculated as follows based on the elemental analysis value.

Here, calculation of the substitution degree when the functional group a is a skeleton of the general formula (III) will be described. As an example, the calculation method will be described by taking as an example the case of the calcium salt “—SO ₃ Ca _1/2 ” when m in the general formula (III) is 0. First, the degree of substitution for uncrosslinked cellulose is calculated by setting the molecular weight of the glucose skeleton, which is one unit of cellulose, to 162. In cellulose, when one primary hydroxyl group is replaced by —SO ₃ Ca _1/2 , that is, when the degree of substitution is 1, the molecular weight is
{(One sulfuric anhydride component) + (Ca _1/2 ) − (hydrogen)} (99)
When the elemental analysis value of S is calculated,
32/261 = about 12%
It becomes. When the degree of substitution is 2, the molecular weight increases to 360, and the elemental analysis value of S is
64/360 = about 17.8%
become. That is, if the elemental analysis value of S is Y and the substitution degree is n,
Y = 32n / (162 + 99n)
Because
n = 162Y / (32-99Y)
It becomes. Using this equation, the substitution degree (n) can be calculated from the elemental analysis value (Y) of S. For example, when the elemental analysis value of S is 18%, it is about 2.1 when converted to the degree of substitution. In the case of cross-linked cellulose, calculation is performed taking into account the degree of cross-linking. For example, when the cross-linking structure is a 2-hydroxy-1,3-ether skeleton obtained from epichlorohydrin, the molecular weight of the glucose skeleton and the cross-linking structure portion and the cross-linking ratio can be calculated.

For example, if the degree of cross-linking is 0.1 according to the definition described later, the molecular weight of the cross-linked structure portion (C ₃ H ₄ O) is 56. Therefore, per glucose skeleton (0.1 × 720 × 56 ÷ 240). =) 16.8. That is, the molecular weight per unit of crosslinked cellulose is (162 + 16.8 =) 179.6. When this is applied to the above-described formula for cellulose, the degree of substitution (n) can be calculated from n = 179.6Y ÷ (32−99Y) and the elemental analysis value (Y) of S.

Next, calculation of the degree of substitution when the functional group a is a skeleton of the general formula (II) will be described. As an example, in the case of the calcium salt “—CH ₂ —COOCa _1/2 ” where l is 0 and n is 0 in the general formula (II), one hydroxyl group of the glucose skeleton which is one unit of cellulose is substituted. As a result, the molecular weight of one unit is 162 + 78 = 240. If the elemental analysis value of Ca is Y and the degree of substitution is n,
Y = 20 ÷ (162 + 78n)
If the elemental analysis value Y of Na is obtained, the substitution degree n is
n = 162Y / (20-80Y)
It can be calculated by

When the functional group a is a skeleton represented by the general formula (IV) and alk is CH ₂ , the degree of substitution can be calculated from the elemental analysis value (Y) of P. That is, in the case of —CH ₂ O—P (O) (OCa _1/2 ) ₂ , assuming that one hydroxyl group of the glucose skeleton, which is one unit of cellulose, is substituted, the molecular weight of one unit is 162 + 148 = 310. .

Y = 31 ÷ (162 + 148n)
If the elemental analysis value Y of P is calculated,
n = 162Y / (31-148Y)
Can be calculated.

  When the functional group a is a skeleton represented by the general formula (V), when the functional group a is (IV) by adding the molecular weight of the alkylene group as in the case of the skeleton of (II) described above What is necessary is just to calculate similarly.

  When there are two or more functional groups, the molecular weight of each functional group portion is calculated, and the degree of substitution may be obtained from the elemental analysis values in the same manner as the calculation method described above.

  In order to set the degree of substitution within the above range, it is preferable to control the degree of crosslinking in the crosslinking step. However, in the conventional sulfation method, it is not easy to obtain a preferable degree of substitution only by controlling the degree of crosslinking. Hereinafter, the case where the functional group a is a skeleton of the general formula (III) will be described as an example.

  In order to improve the substitution degree of the sulfate group, the amount ratio of the sulfating agent to the crosslinked cellulose may be increased. However, since the anhydrous sulfuric acid / DMF complex, which is the most powerful sulfating agent, has a solubility of sulfuric anhydride of less than 20% in DMF, the concentration of the sulfating agent in the reaction system is naturally limited. As a result, there is a limit in improving the degree of substitution, and it has been difficult to increase the degree of substitution especially for crosslinked cellulose having a high degree of crosslinking.

  Regarding this point, by adding sulfuric anhydride directly into a reaction system in which crosslinked cellulose is dispersed in a DMF solvent, the concentration of anhydrous sulfuric acid in the DMF is constant, but the concentration relative to the crosslinked cellulose is about twice or more. Accordingly, even when the ratio of anhydrous sulfuric acid / cellulose is about 2/1, it is possible to easily obtain a substitution degree (1.5 or more) of the same degree or more as in the conventional case of 3/1. I found out. That is, in order to obtain a high degree of substitution for a crosslinked cellulose having a high degree of crosslinking, this can be achieved by reducing the amount of DMF with respect to the crosslinked cellulose and reacting sulfuric anhydride in a high concentration state.

  The obtained metal salt of the crosslinked cellulose derivative may be used as it is, but may be further purified by alcohol precipitation, ion exchange resin chromatography, gel filtration chromatography or the like, if necessary.

  The metal salt of a crosslinked cellulose derivative is useful as a uremic ameliorating drug that can be orally administered to the human body. In particular, the period until the next dialysis of a renal failure patient undergoing hemodialysis treatment can be extended, and the burden on the patient can be greatly improved.

  As will be described later in the <Evaluation Test> of the Examples, the metal salt of the crosslinked cellulose derivative has an effect of extending the survival time of the total nephrectomized rat. This excellent life-prolonging effect is presumed to be due to the fact that the metal salt of the crosslinked cellulose derivative traps the causative substance of uremia secreted from the digestive tract or produced in the intestinal tract.

  The metal salt of the crosslinked cellulose derivative can be contained in a uremic drug, various foods, and the like.

  The uremic remedy containing a metal salt of a crosslinked cellulose derivative can be produced by processing the metal salt obtained by the above method into various forms by a usual method. For example, solid, liquid, emulsion, paste, jelly and the like.

  The metal salt of a crosslinked cellulose derivative can be effectively applied to food. Foods containing these metal salts include any of those that can be eaten immediately, those that are cooked and eaten, premixed materials for food production, and the like. The solid product may be any of powder, granule, and solid, such as confectionery such as biscuits, cookies, cakes, snacks, rice crackers, bread, and powdered beverages (powdered coffee, cocoa, etc.). included. Examples of liquids, emulsions, pastes, and jellys include various beverages such as juices, carbonated beverages, and lactic acid bacteria beverages.

  Examples of dosage forms of uremia-improving drugs include tablets, powders, granules, fine granules, liquids, etc. These preparations are formulated according to a conventional method with a metal salt of a crosslinked cellulose derivative together with a pharmaceutically acceptable carrier. Can be manufactured.

  The appropriate amount of the metal salt of the crosslinked cellulose derivative varies depending on the degree of renal failure, but it is preferable to take about 0.5 to 50 g per day.

  Hereinafter, the present invention will be described in more detail with reference to examples, but the present invention is not limited thereto.

[Reference production example: Production of sulfated cellulose calcium salt: CaCS-005 (Lot.IK031215)]
{First stage: Sulfation process}
20.0 g of crystalline cellulose (made by Asahi Kasei, trade name: Theolas PH-101) vacuum-dried at 40 ° C. was put into a 500 mL separable flask, and impregnated with 100 mL of DMF with stirring for 4 days.

The suspension was cooled to 5 ° C. Next, with stirring, 371.8 g of 20% sulfuric anhydride-DMF complex solution (SO ₃ : 0.929 mol) was gradually added dropwise from a dropping funnel connected to the separable flask while maintaining the system temperature at 5 ° C. After completion of dropping, the temperature was adjusted to 16 to 17 ° C. in a thermostatic bath and stirred for 6 hours.

  Next, 500 mL of isopropanol was added to the reaction solution. The precipitate from the reaction solution was filtered off. The filtrated product was dissolved in 500 mL of water, 137.0 g of a saturated aqueous solution of calcium chloride was added to perform calciumation, and the crude product was precipitated and filtered to obtain the desired product. Further, it was washed with isopropanol (500 mL × 2 times). After isopropanol was removed by filtration, it was dried in a vacuum dryer to obtain 27.5 g of sulfated cellulose calcium salt. Upon elemental analysis, the following was found: S; 16.0%, Ca; 12.4%. The degree of substitution was 1.6.

{Second stage: UF membrane ultrafiltration process}
25.05 g of the sulfated cellulose calcium salt obtained in the first stage was dissolved in 2000 mL of ion-exchanged water with cooling.

  Next, ultrafiltration was performed at a flow rate of 1.88 to 1.92 L / min using a UF membrane (manufactured by Asahi Kasei: pencil type module ACP-0013, nominal molecular weight cut off: 13,000). Circulating and concentrated until the stock solution reached 200 mL. 200 mL portion was freeze-dried to finally obtain 17.68 g of white powder.

  Upon elemental analysis, the following was found: S; 18.3%, Ca; 12.4%. The degree of substitution was 2.1.

[Production example: Production of crosslinked sulfated cellulose calcium salt: CaCS-006 (Lot.IK031224)]
{First stage; Crosslinking process}
Crystalline cellulose (manufactured by Asahi Kasei, trade name: Theolas PH-101) 5.0 g (30.8 mmol (in terms of glucose)) was placed in a 500 mL three-necked flask, and a condenser tube and a dropping funnel were connected. Separately, 8.6 g (206 mmol) of sodium hydroxide (96% product) was dissolved in 50 mL of water. The prepared aqueous sodium hydroxide solution was transferred to a dropping funnel and added to crystalline cellulose. The mixture was stirred at room temperature for 10 minutes to maintain a suspended state.

  Next, 50 mL of n-heptane and 50 mL of methanol were added to the reaction system, and the temperature was raised to 50 ° C. A dropping funnel was used to rapidly drop 8.5 g (92 mmol) of epichlorohydrin dissolved in 50 mL of methanol into the suspension. While mixing the two layers well, the mixture was stirred for 3 hours while maintaining 50-60 ° C.

  After cooling to room temperature, concentrated hydrochloric acid was gradually added with stirring, and the pH of the aqueous layer was adjusted to around 7.0. Once filtered under reduced pressure, the filtrate is washed with water and further with methanol. After removing methanol by filtration, it was sufficiently dried by a vacuum dryer to obtain 6.1 g of crosslinked cellulose. The degree of crosslinking was 0.174.

{Second stage; Sulfation process}
4 g (0.0247 mol (in terms of glucose)) of the crosslinked cellulose obtained in the first stage was placed in a 500 mL separable flask and impregnated with 20 mL of DMF with stirring for 3 days. The suspension was then cooled to 5 ° C. Next, with stirring, 74.12 g of 20% sulfuric anhydride-DMF complex solution (SO ₃ : 0.185 mol) was gradually added dropwise from a dropping funnel connected to the separable flask. At this time, the reaction temperature was maintained at 5 ° C. After completion of dropping, the temperature was adjusted to 16 to 17 ° C. in a thermostatic bath and stirred for 24 hours.

  Next, 100 mL of isopropanol was added to the reaction solution. The precipitate from the reaction solution is separated by filtration. The filtrate was added with 250 mL of water and stirred. Furthermore, 27.45 g of a saturated calcium chloride aqueous solution was added and stirred for 2 hours. The insoluble material was filtered off and further washed with isopropanol (100 mL × 2 times). After isopropanol was removed by filtration, the product was sufficiently dried in a vacuum dryer to obtain 8.09 g of a crosslinked sulfated cellulose calcium salt. Upon elemental analysis, the result was S; 17.6%. The degree of substitution was 2.0.

[Production Example: Production of crosslinked sulfated cellulose calcium salt: CaCS-007 (Lot.IK040113)]
{First stage; Crosslinking process}
16.0 g (99 mmol (converted to glucose)) of crystalline cellulose (manufactured by Asahi Kasei, trade name: Theorus PH-101) was placed in a 500 mL three-necked flask, and a condenser tube and a dropping funnel were connected. Next, 300 mL of n-heptane and 10 mL of isopropanol were added to the reaction system. After stirring for 30 minutes, 32.0 g (768 mmol) of sodium hydroxide (96% product) was separately dissolved in 100 mL of water. Half of the prepared sodium hydroxide solution was transferred to a dropping funnel, added to the suspension, and stirred at room temperature for 1 hour.

  Using a dropping funnel, 26.0 g (281 mmol) of epichlorohydrin was quickly added dropwise to the suspension, and the temperature was raised to 50 ° C. While mixing the two layers well, the mixture was stirred for 3 hours while maintaining 50-60 ° C. Next, the remaining sodium hydroxide solution was added dropwise and stirred for 1.5 hours. To this was added 26.0 g (281 mmol) of epichlorohydrin, and the mixture was further stirred for 2 hours.

  After cooling to room temperature, insoluble matters were filtered off, and the filtrate was washed successively with water, 2N hydrochloric acid, and water until the pH of the wash became near neutral. Further, it was washed with methanol. It was dried with a vacuum dryer to obtain 18.1 g of crosslinked cellulose. The degree of crosslinking was 0.198.

{Second stage; Sulfation process}
10 g (0.0617 mol (in terms of glucose)) of the crosslinked cellulose obtained in the first stage was placed in a 500 mL separable flask and impregnated with 50 mL of DMF for 3 days with stirring. The suspension was then cooled to 5 ° C. Next, with stirring, 206.04 g (SO ₃ : 0.463 mol) of 18% anhydrous sulfuric acid-DMF complex solution was gradually added dropwise from a dropping funnel connected to the separable flask. At this time, the reaction temperature was maintained at 5 ° C. After completion of dropping, the temperature was adjusted to 16 to 17 ° C. in a thermostatic bath and stirred for 24 hours.

  Next, 250 mL of isopropanol was added to the reaction solution. The precipitate from the reaction solution is separated by filtration. The filtrate was added with 625 mL of water and stirred. Furthermore, 68.60 g of saturated calcium chloride aqueous solution was added and stirred for 2 hours. The insoluble material was filtered off and further washed with isopropanol (100 mL × 2 times). After isopropanol was removed by filtration, the product was sufficiently dried in a vacuum drier to obtain 13.7 g of a crosslinked sulfated cellulose calcium salt. Upon elemental analysis, the following was found: S; 7.40%, Ca; 5.38%. The degree of substitution was 0.85.

[Production Example: Production of crosslinked sulfated cellulose calcium salt: Lot.IK-40223]
{First stage; Crosslinking process}
Crystalline cellulose (manufactured by Asahi Kasei, trade name: Theolas PH-101) 80.0 g was placed in a 3 L three-necked flask and suspended in 800 mL of MeOH and 800 mL of n-hexane. A condenser, a stirring blade (with a stirring motor), and a dropping funnel were connected and stirred. Separately, 140.0 g (3.36 mol) of sodium hydroxide (96% product) is dissolved in 1600 mL of water. The prepared aqueous sodium hydroxide solution was transferred to a dropping funnel and added to the suspension. Stir for 30 minutes at room temperature to maintain suspension.

  Next, the temperature of the reaction system was raised to 50 ° C., and a solution of 138.8 g (1.5 mol) of epichlorohydrin in 200 mL of methanol was dropped into the suspension using a dropping funnel. While mixing the two layers well, the mixture was stirred for 3 hours while maintaining 50-60 ° C.

  After cooling to room temperature, the mixture was washed successively with water, 2N hydrochloric acid and water until the washing solution reached pH = 7.0.

  The filtrate was filtered under reduced pressure, and the filtrate was washed with methanol. Methanol was removed by filtration, followed by drying with a vacuum dryer to obtain 84.4 g of crosslinked cellulose. The degree of crosslinking was 0.053.

{Second stage; Sulfation process}
40 g (0.247 mol) of the crosslinked cellulose obtained in the first stage was placed in a 500 mL separable flask and impregnated with 200 mL of DMF for 3 days with stirring. The suspension is then cooled to 5 ° C. Next, with stirring, 900 g of 18% sulfuric anhydride-DMF complex solution (SO ₃ : 2.025 mol) is gradually added dropwise from a dropping funnel connected to the separable flask. At this time, the reaction temperature is maintained at 5 ° C. After completion of dropping, the temperature is adjusted to 16 to 17 ° C. in a thermostatic bath, and the whole is stirred overnight.

  Next, 1000 mL of isopropanol is added to the reaction. The precipitate from the reaction solution is separated by filtration. Dissolve the filtrate in 1000 mL of water, add 274.5 g of a saturated aqueous solution of calcium chloride to perform calciumation, precipitate the crude product, and separate by filtration to obtain the desired product. Further, it was washed with isopropanol (1000 mL × 2 times). After isopropanol was removed by filtration, the product was sufficiently dried in a vacuum dryer to obtain 76.8 g of a crosslinked sulfated cellulose calcium salt. Upon elemental analysis, the following was found: S; 16.8%, Ca; 12.8%. The degree of substitution was 1.9.

[Production example: Production of crosslinked sulfated cellulose calcium salt: Lot.SS-1054]
{First stage; Crosslinking process}
Crystalline cellulose (made by Asahi Kasei, trade name: Theolas PH-101) 80.0 g is put into a 3 L three-necked flask, and a condenser, a stirring blade (with a stirring motor), and a dropping funnel are connected. Thereto, 800 mL of methanol and 800 mL of n-hexane are added and stirred to form a suspension. Separately, 144.0 g (3.457 mol) of sodium hydroxide (96% product) is dissolved in 800 mL of water under ice cooling. The prepared aqueous sodium hydroxide solution is transferred to a dropping funnel and added to the suspension. Stir at room temperature for 60 minutes to maintain suspension.

  Next, the temperature of the reaction system is raised to 50 ° C., and using a dropping funnel, 159.9 g (1.728 mol) of epichlorohydrin dissolved in 200 mL of methanol is quickly added dropwise to the suspension. The mixture is stirred for 6 hours while maintaining 50-60 ° C. so that the two layers are well mixed.

  Cool to room temperature and continue stirring for another 20 hours, then transfer to a 5 L beaker and let stand for 4 hours. The supernatant is removed by decantation, and the residue is suspended in water and filtered under reduced pressure. The filtrate is suspended again with water, and concentrated hydrochloric acid is gradually added while stirring to bring the pH to around 7.0. Once filtered under reduced pressure, the filtrate is washed with water and further with methanol. After removing methanol by filtration, the product was sufficiently dried by a vacuum dryer to obtain 80.8 g of crosslinked cellulose. The degree of cross-linking was less than 0.01.

{Second stage; Sulfation process}
30 g of the crosslinked cellulose obtained in the first step was placed in a 2 L three-necked flask, 150 mL of DMF was added, and the mixture was stirred at room temperature for 21 hours. The suspension is cooled to 5 ° C. Next, with stirring, 721.0 g (0.871 mol) of anhydrous DMF complex DMF solution (18.5% product) is gradually added dropwise from a dropping funnel connected to the three-necked flask. At this time, the reaction temperature is maintained at 5 ± 5 ° C. After completion of dropping, the temperature is raised to room temperature and stirred for 24 hours.

  Next, pressure filtration is performed using 1000 mL of isopropanol as a cleaning liquid. After the filtrate was dissolved in 750 mL of water, 198.0 g (0.555 mol) of a calcium chloride aqueous solution (31.1%) prepared separately was added. While stirring, 1000 mL of isopropanol was added to precipitate crystals, and the mixture was allowed to stand for 60 minutes. The supernatant is removed by decantation, and the residue is suspended in isopropanol and filtered under reduced pressure. Further, it was washed with isopropanol. After isopropanol was removed by filtration, the product was sufficiently dried with a vacuum drier to obtain 79.8 g of a crosslinked sulfated cellulose calcium salt. Upon elemental analysis, the following was found: S; 17.8%, Ca; 12.4%. The degree of substitution was 2.0.

[Production Example: Production of crosslinked sulfated cellulose calcium salt: Lot. Type-007]
{First stage; Crosslinking process}
Crystalline cellulose (made by Asahi Kasei, trade name: Theolas PH-101) 80.0 g is put into a 3 L three-necked flask, and a condenser, a stirring blade (with a stirring motor), and a dropping funnel are connected. Separately, 329.2 g (7.901 mol) of sodium hydroxide (96% product) is dissolved in 800 mL of water. This prepared aqueous sodium hydroxide solution is transferred to a dropping funnel and added to crystalline cellulose. Stir at room temperature for 30 minutes to maintain suspension.

  Next, the temperature of the reaction system is raised to 50 ° C., and a solution of 228.4 g (2.469 mol) of epichlorohydrin in 1,600 mL of n-heptane is quickly added dropwise to the suspension using a dropping funnel. The mixture is stirred for 3 hours while maintaining 50-60 ° C. so that the two layers are well mixed.

  After cooling to room temperature, concentrated hydrochloric acid is gradually added while stirring to bring the pH of the aqueous layer to around 7.0. Once filtered under reduced pressure, the filtrate is washed with water and further with methanol. After removing methanol by filtration, it was sufficiently dried by a vacuum dryer to obtain 94.9 g of crosslinked cellulose. It was confirmed from the chart showing that the crystallinity obtained by X-ray diffraction of this compound was lost, that the crosslinking was advanced. The degree of crosslinking was 0.180.

{Second stage; Sulfation process}
25 g of the crosslinked cellulose obtained in the first step was placed in a 500 mL three-necked flask together with calcium chloride and 34.3 g (0.309 mol), and DMF and 300 mL were added and stirred at room temperature for 24 hours. The suspension is cooled to 5 ° C. Next, with stirring, 28.9 mL (0.694 mol) of anhydrous sulfuric acid is gradually added dropwise from a dropping funnel connected to the three-necked flask. At this time, the reaction temperature is maintained at 20 ± 5 ° C. After completion of dropping, the temperature is raised to room temperature and stirred for 24 hours.

  Next, it is filtered under reduced pressure using 1000 mL of isopropanol as a washing solution. Wash the filtrate with water (1000 mL x 3 times). Further, it was washed with methanol (500 mL × 2 times). After removing methanol by filtration, it was sufficiently dried with a vacuum dryer to obtain 48.0 g of a crosslinked sulfated cellulose calcium salt. Upon elemental analysis, the following was found: S; 16.7%, Ca; 9.7%. The degree of substitution was 1.9.

[Production Example: Production of crosslinked sulfated cellulose calcium salt: Lot. Type-007-B]
{First stage; Crosslinking process}
After adding 18.02 kg of water and 13.71 kg of n-heptane to a 100 L-GL reactor equipped with a condenser, stirrer, thermometer and dropping tank, cool to 5 ° C. Dissolve 8.20 kg (0.1968 kmol) of sodium hydroxide (96% product) and raise the temperature to 20 ° C. Separately, 1.998 kg of crystalline cellulose (manufactured by Asahi Kasei, trade name: Theolas PH-101) is suspended in 6.80 kg of n-heptane. This suspension is added to the prepared aqueous sodium hydroxide solution, and 6.80 kg of n-heptane and 2.0 kg of water are further added, followed by stirring at room temperature for 30 minutes to maintain the suspended state.

  Next, the temperature of the reaction system is raised to 55 ° C., and 5.70 kg (61.73 mol) of epichlorohydrin is quickly added dropwise thereto using a dropping tank. The mixture is stirred for 3 hours while maintaining 50-60 ° C. so that the two layers are well mixed.

  After cooling to room temperature, concentrated hydrochloric acid is gradually added while stirring to bring the pH of the aqueous layer to around 7.0. Once filtered under pressure, the filtrate is washed with water and then with methanol. After removing methanol by filtration, it was sufficiently dried by a vacuum dryer to obtain 1.91 kg of crosslinked cellulose. It was confirmed from the chart showing that the crystallinity obtained by X-ray diffraction of this compound was lost, that the crosslinking was advanced. The degree of crosslinking was 0.180.

{Second stage; Sulfation process}
219.7 g of the crosslinked cellulose obtained in the first stage was placed in a 5 L separable flask, and 3520 mL of DMF was added under a nitrogen stream to obtain a suspension. After further cooling to 5 ° C, add 301.5 g (2.72 mol) of calcium chloride. The mixture was warmed to room temperature and stirred for 23 hours under a nitrogen stream. The suspension is cooled to 5 ° C. Next, with stirring, 254.6 mL (6.11 mol) of anhydrous sulfuric acid is gradually added dropwise from a dropping funnel connected to the separable flask. At this time, the reaction temperature is maintained at -20 to 0 ° C. After completion of dropping, the temperature is raised to room temperature and stirred for 24 hours.

  Next, 1400 mL of isopropanol is added to the reaction system and stirred for 10 minutes, followed by pressure filtration. Wash with 3000 mL of isopropanol, add the filtrate to 3000 mL of water, and stir to make a suspension. Calcium hydroxide is gradually added to this suspension to bring the pH of the aqueous layer to around 7.0. The mixture was filtered under pressure and further washed with water (1600 mL × 4 times) to obtain 1994.4 g of a filtrate. 1894.4g of this filtrate is washed with denatured ethanol (manufactured by Nippon Alcohol Sales, trade name: Solmix AP-7) (1894.4g x 2) and water (1894.4g x 2), and then sufficiently with a vacuum dryer Dried. Grinding with a mill and sieving (opening: 250 μm) were performed, and 463.2 g of a crosslinked sulfated cellulose calcium salt was obtained again by drying with a vacuum dryer. Upon elemental analysis, the following was found: S; 13.8%, Ca; 9.8%. The degree of substitution was 1.5.

<Evaluation test>
{Effects of cross-linked cellulose sulfate calcium salt on survival time of total nephrectomized rats: Lot. Type007-B}
〔Method〕
The animals used were 8-week-old SD male rats purchased from Nippon Charles River Co., Ltd. On the first day of the test, incisions were made on the left and right skin and muscle layers at the lower part of the dorsal ribs of the rat, and the capsule and adrenal glands were removed from each kidney and pulled out of the body. After that, the incision was sutured. These surgeries were performed under ether anesthesia. The test substance was suspended in distilled water at 14:00 and 20:00 on the day of creation of the nephrectomized rat, and the following day (test day 2) at 8:00, 14:00, and 20:00. And 1.0 g / kg were administered by oral sonde. Distilled water was administered to the control group at 10 mL / kg. Feeding and watering were given freely until 8:00 am on the third day of the test, but after that, the animals were raised under fasting and fasting conditions. Survival was observed every 4 hours on the first day of the test at 12:00 p.m. (0 hour) on the first day of the test until every 96 hours.

〔result〕
FIG. 1 shows the survival curve of each group. In FIG. 1, Water is distilled water in the control group, 0.1, 0.3, and 1.0 g / kg are doses of the crosslinked cellulose sulfate calcium salt administered to each group at a time. The number of animals was shown.

  In the control group, all cases died at 76 hours. At this time, 1 case was alive in the 0.1 g / kg group, 11 cases were in the 0.3 g / kg group, and 14 cases were alive in the 1 g / kg group. At 84 hours, all deaths in the 0.1 g / kg group were confirmed. On the other hand, in the 0.3 and 1.0 g / kg groups, 3 cases and 10 cases were alive at 96 hours, respectively, and a clear extension of the survival time was confirmed.

<Safety test 1>
{Effect of cross-linked sulfated cellulose calcium salt on the digestive tract of normal rats 1}
〔Method〕
The animals were grouped using 7-9 week old CBA male mice purchased from Nippon Charles River Co., Ltd. using body weight as an index. In each group of mice, 10% of the commercially available powdered feed (Oriental Yeast Co., Ltd., powder CRF1) was replaced with cellulose (powder, Nacalai Tesque) or test substance (SS1054, IK-40223, Type007). The test meal was allowed to eat freely for one week. At least one stool sample is collected from each mouse after the intake of the test food, and a fecal occult blood test (Stool occult blood slide Shionogi II, Shionogi Pharmaceutical Co., Ltd.) is performed using a slide, depending on the degree of color change. The scores were scored from 0 (no change) to 5 (dark).

〔result〕
All cases in the cellulose group had normal stool properties, and the fecal occult blood reaction score was 0.5 ± 0.0 (n = 8, mean ± standard deviation). In the SS-1054 group, gastrointestinal symptoms worsened as blood deposition around the anus was observed in 6 out of 8 cases, and systemic symptoms became more severe as the amount of food consumption decreased after the 4th day of the study. It became serious. In the IK-40223 group, there was no decrease in food consumption during the 7-day study period, and the fecal occult blood test score was 1.5 ± 0.8 (8). In the Type007 group, the stool of all mice was normal, and the fecal occult blood reaction score was 0.5 ± 0.3 (10).

<Safety test 2>
{Effect of cross-linked sulfated cellulose calcium salt on the digestive tract of normal rats 2}
〔Method〕
The animals were grouped using body weight as an index using 9-week-old male CBA mice purchased from Charles River Japan. In each group of mice, 10% of commercially available powdered feed (Oriental Yeast Co., Ltd., powder CRF1) was replaced with cellulose (powder, Nacalai Tesque) or test substances (CaCS005, CaCS006, CaCS007). The food was ingested freely for 6 days. At least one stool sample is collected from each mouse after the intake of the test food, and a fecal occult blood test (Stool occult blood slide Shionogi II, Shionogi Pharmaceutical Co., Ltd.) is performed using a slide, depending on the degree of color change. The scores were scored from 0 (no change) to 5 (dark).

〔result〕
All cases in the cellulose group had normal stool properties, and the fecal occult blood reaction score at the end of the test was 0.3 ± 0.3 (n = 5, mean ± standard deviation). In the CaCS005 group, fecal occult blood reaction (score 1.8 ± 0.5 (n = 4)) was observed from the second day of the study. Since then, there was a marked decrease in food consumption in the animals of the CaCS005 group, and there were animals that became more severe as mucous stool was seen, and in all cases the limbs were whitened due to anemia. The fecal occult blood reaction score at the end of the study was 3.3 ± 1.3 (n = 4). The fecal characteristics of the CaCS006 group and CaCS007 group remained almost normal during the study period, and the score at the end of the study was 0.5 ± 0.0 in the CaCS006 group (n = 3, but on the fifth day of the study), 0.2 ± 0.3 in the CaCS007 group (N = 3) Obviously the damage to the gastrointestinal tract was reduced. There was no anemia.

It is the figure which showed the survival time of the nephrectomy rat under bridge | crosslinking sulfated cellulose calcium salt administration in <evaluation test>.

Claims (10)

A drug for improving uremia, comprising a metal salt of a crosslinked cellulose derivative represented by the following general formula (I) as an active ingredient.
R-O-A (I)
{In Formula (I), R represents a crosslinked cellulose residue, and A represents a functional group a having a cation exchange ability. }   The uremic disease-improving drug according to claim 1, wherein the degree of substitution of the hydroxyl group of the glucose unit of the crosslinked cellulose derivative with the functional group a is 1.2 or more.   The uremic disease-improving drug according to claim 1, wherein the degree of substitution of the hydroxyl group of the glucose unit of the crosslinked cellulose derivative with the functional group a is 1.4 or more.   The uremic remedy according to claim 1, wherein the degree of substitution of the hydroxyl group of the glucose unit of the crosslinked cellulose derivative with the functional group a is 1.5 or more. The functional group a is selected from the groups represented by the following general formulas (II) to (V), The uremic disease-improving drug according to any one of claims 1 to 4.

{In Formulas (II) to (V), alk represents an alkylene group having 1 to 6 carbon atoms, l represents an integer of 0 to 5, m represents 0 or 1, and n represents an integer of 0 to 2. To express. } The functional group a is represented by the following general formulas (II-1), (II-2), (III-1) to (III-5), (IV-1), (V-1) and (V-2). 6. The uremic agent for improving uremia according to claim 5, which is selected.

The combination of a plurality of types of functional groups a is any one of the following (c-1) to (c-3), and the uremia-improving drug according to claim 6.

  The uremic disease-improving drug according to any one of claims 1 to 7, wherein the crosslinked cellulose derivative is produced using crystalline cellulose.   9. The uremia-improving drug according to any one of claims 1 to 8, wherein the crosslinked cellulose derivative is produced using epichlorohydrin as a crosslinking agent.   A food for improving uremia, comprising the metal salt of the crosslinked cellulose derivative according to any one of claims 1 to 9.

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