WO2019093505A1 - Composition pour empêcher les adhérences - Google Patents

Composition pour empêcher les adhérences Download PDF

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WO2019093505A1
WO2019093505A1 PCT/JP2018/041770 JP2018041770W WO2019093505A1 WO 2019093505 A1 WO2019093505 A1 WO 2019093505A1 JP 2018041770 W JP2018041770 W JP 2018041770W WO 2019093505 A1 WO2019093505 A1 WO 2019093505A1
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layer
alginic acid
metal salt
sponge
monovalent metal
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PCT/JP2018/041770
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English (en)
Japanese (ja)
Inventor
大知 伊藤
誠一 太田
長谷川　潔
三津子 伊佐次
賢 清水
Original Assignee
持田製薬株式会社
国立大学法人東京大学
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Priority to JP2019552417A priority Critical patent/JP7267204B2/ja
Publication of WO2019093505A1 publication Critical patent/WO2019093505A1/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
    • A61L31/04—Macromolecular materials
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
    • A61L31/12—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
    • A61L31/14—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
    • A61L31/14—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L31/16—Biologically active materials, e.g. therapeutic substances

Definitions

  • the present invention relates to an anti-adhesion material, a method for producing the same, a sponge-like laminate and a pharmaceutical composition.
  • Adhesion refers to a state in which surfaces of tissues to be separated from one another are connected or fused with fibrous tissue. Adhesions are caused by an exudate containing fibrin on the surface of tissue accompanying trauma or inflammation, and the exudate is organized to connect or fuse tissue surfaces.
  • the trauma that can be caused on the surface of tissue in surgery, the inflammation caused by trauma, and the inflammation caused by the drying of the tissue surface in surgery cause adhesion to occur.
  • Adhesions can sometimes cause infertility, intestinal transit problems, chronic pelvic pain. Also, another surgical procedure may be necessary to ablate adhesions that have developed after surgery. For example, although multiple operations are effective for recurrent cases of liver cancer, it is possible to determine whether to apply re-operation, the risk of treatment, the amount of bleeding at the time of operation, etc. It largely depends on the prevention of adhesions later. From these things, it is necessary to prevent adhesions, and various measures have been taken so far to prevent adhesions.
  • Some such means for adhesion prevention are to provide a physical barrier that is placed between the site of trauma or inflammation and its adjacent tissue to prevent tissue joining or fusion. Sheet-like ones are known as such physical barriers.
  • the sheet-like material includes polytetrafluoroethylene (PTFE) film (Preclude (trade name) (WL Gore and Associates, Inc.)), hyaluronic acid (HA) and carboxymethylcellulose (CMC). Sheets (Seprafilm (trade name) (Genzyme GmbH)), regenerated oxidized cellulose sheets (INTERCEED (trade name) (Johnson & Johnson)), and the like.
  • PTFE polytetrafluoroethylene
  • Preclude (trade name) (WL Gore and Associates, Inc.)
  • HA hyaluronic acid
  • CMC carboxymethylcellulose
  • Sheets Seprafilm (trade name) (Genzyme GmbH)
  • INTERCEED regenerated oxidized cellulose sheets
  • the PTFE film is not biodegradable and thus has the problem of remaining in the body.
  • the sheet containing HA and CMC and regenerated oxidized cellulose sheet are biodegradable, they can not completely prevent serious adhesions such as adhesions that occur after hepatectomy, and there is room for improvement in terms of the adhesion prevention effect. was there.
  • a biocompatible material selected from proteins such as collagen and polysaccharides such as carboxymethylcellulose, hyaluronic acid and alginic acid is made into a sheet form and a particulate form to be used as a medical absorbent, medical patch, adhesion preventive material It is known to use as a living tissue reinforcing material etc. (Patent Documents 1 to 8). It is also known to incorporate drugs into such biocompatible materials (US Pat. Nos. 6,798,839 and 6,087,015).
  • adhesion-preventing effect is high, both adhesion in wounds and de novo adhesions can be suppressed, not adversely affecting the applied body, not disturbing wound healing, for intestinal anastomosis etc.
  • an anti-adhesion material having at least one performance that can be used, is easy to apply via a trocker in endoscopic surgery, is capable of adjusting and re-adhering a sticking position, and the like.
  • an adhesion-preventing material which combines the advantages of a film (sheet) -like adhesion-preventing material and the advantages of a spray (liquid / gel) adhesion-preventing material
  • a sponge-like anti-adhesion material applicable to a living body having different dissolution rates of the first layer and the second layer, specifically, alginic acid having a relatively high weight-average molecular weight.
  • Adhesion preventing material including a sponge-like laminate applicable to a living body including a sponge-like second layer, and adhesion not only for prevention of adhesion for a local operation but also adhesion in a wide range of application area It found such that it has a locking effect.
  • the present inventors have selected at least one layer selected from the first layer and the second layer of the sponge-like laminate from the group consisting of drugs, growth factors, hormones, proteins and combinations thereof. It has been found that an anti-adhesion material containing at least one pharmaceutical ingredient has an even better anti-adhesion effect. Based on these findings, the present inventors have completed the present invention.
  • the present invention is as follows. [1-1] The weight of the monovalent metal salt of alginic acid of the first layer, comprising a sponge-like first layer and a second layer of a monovalent metal salt of alginic acid at least partially crosslinked with a curing agent And the weight average molecular weight of the monovalent metal salt of alginic acid in the second layer is 1,000 to 1,000,000, and the weight average molecular weight is decrosslinking.
  • the weight average molecular weight of the monovalent metal salt of alginic acid in the first layer is higher than the weight average molecular weight of the monovalent metal salt of alginic acid in the second layer
  • At least one layer selected from the one layer and the second layer comprises at least one pharmaceutical component selected from the group consisting of a drug, a growth factor, a hormone, a protein and a combination thereof, and is sterilized Living body Anti-adhesion material, including sponge-like laminates applicable to [1-1a]
  • a monovalent metal salt of alginic acid of a first layer comprising a sponge-like first layer and a second layer of a monovalent metal salt of low endotoxin alginic acid crosslinked at least partially with a curing agent
  • the weight average molecular weight of the monovalent metal salt of alginic acid of the second layer is 1,000 to 1,000,000, and the weight average molecular weight is The weight average molecular weight of the monovalent metal salt of alginic acid in the first layer is higher than the weight average molecular weight of the monovalent metal salt of alginic acid in the second layer, as measured by the GPC-MALS method after the decrosslinking treatment.
  • At least one layer selected from the first layer and the second layer comprises at least one pharmaceutical component selected from the group consisting of drugs, growth factors, hormones, proteins and combinations thereof
  • An anti-adhesion material comprising a sterile, bio-compatible sponge-like laminate.
  • the total amount of the monovalent metal salt of alginic acid (eg, monovalent metal salt of low endotoxin alginic acid) in the first and second layers is 0.1 mg / cm 2 to 3 mg / mg
  • Any one of the above [1-1] to [1-4], wherein the endotoxin content of the monovalent metal salt of alginic acid in the first layer and the second layer is 500 EU / g or less
  • the anti-adhesion material described in the item is 0.1 mg / cm 2 to 3 mg / mg
  • the monovalent metal salt of alginic acid of the first layer and the second layer is sodium alginate or potassium alginate described in any one of the above [1-1] to [1-5] Anti-adhesion material.
  • the curing agent of the first layer and the second layer is at least one metal ion compound selected from the group consisting of CaCl 2 , CaSO 4 , ZnCl 2 , SrCl 2 , FeCl 3 and BaCl 2.
  • a biologically applicable sponge-like laminate comprising a first layer and a second layer each containing a monovalent metal salt of alginic acid at least partially cross-linked with a curing agent; The dissolution rate of the layer is slower than that of the second layer,
  • An adhesion prevention material wherein at least one layer selected from the first layer and the second layer contains at least one pharmaceutical component selected from the group consisting of drugs, growth factors, hormones, proteins and combinations thereof .
  • a biologically applicable sponge-like laminate including a first layer and a second layer each containing a monovalent metal salt of low endotoxin alginic acid crosslinked at least partially with a curing agent, The dissolution rate of one layer is slower than that of the second layer, An adhesion prevention material, wherein at least one layer selected from the first layer and the second layer contains at least one pharmaceutical component selected from the group consisting of drugs, growth factors, hormones, proteins and combinations thereof .
  • the elution amount of the monovalent metal salt of alginic acid in the second layer is 100%.
  • the ratio of the elution amount of the monovalent metal salt of alginic acid in the first layer is less than 50% at 1 hour from the start of measurement, and less than 70% at 2 hour, The adhesion preventing material according to any one of [1-10b].
  • the first layer contains 25 ⁇ 10% by weight of monovalent metal salt of alginic acid Elution occurs within an hour, 80 ⁇ 10% by weight elutes within 4 hours, and the second layer is an elution of 70 ⁇ 10% by weight of monovalent metal salt of alginic acid within 1 hour, 90 ⁇ 10%.
  • the adhesion preventing material according to any one of the above [1-10] to [1-10b] which is eluted and eluted at 80 ⁇ 20% by weight within 4 hours.
  • the weight of the monovalent metal salt of alginic acid of the first layer comprising a sponge-like first layer and a second layer of a monovalent metal salt of alginic acid at least partially crosslinked with a curing agent
  • the weight average molecular weight of the monovalent metal salt of alginic acid in the second layer is 1,000 to 1,000,000, and the weight average molecular weight is decrosslinking.
  • the weight average molecular weight of the monovalent metal salt of alginic acid in the first layer is higher than the weight average molecular weight of the monovalent metal salt of alginic acid in the second layer
  • At least one layer selected from the one layer and the second layer comprises at least one pharmaceutical component selected from the group consisting of a drug, a growth factor, a hormone, a protein and a combination thereof, and is sterilized Living body
  • a method for preventing adhesion comprising applying a sponge-like laminate applicable to the above to a subject requiring adhesion prevention, with the first layer directed to the surface on the wound side.
  • a monovalent metal salt of alginic acid of the first layer comprising a sponge-like first layer and a second layer of a monovalent metal salt of low endotoxin alginic acid crosslinked at least partially with a curing agent
  • the weight average molecular weight of the monovalent metal salt of alginic acid of the second layer is 1,000 to 1,000,000, and the weight average molecular weight is The weight average molecular weight of the monovalent metal salt of alginic acid in the first layer is higher than the weight average molecular weight of the monovalent metal salt of alginic acid in the second layer, as measured by the GPC-MALS method after the decrosslinking treatment.
  • At least one layer selected from the first layer and the second layer comprises at least one pharmaceutical component selected from the group consisting of drugs, growth factors, hormones, proteins and combinations thereof
  • a method for preventing adhesion comprising applying a sterile, organism-applicable sponge-like laminate to a wound-side surface of a subject requiring adhesion prevention, to a first layer.
  • the total amount of the monovalent metal salt of alginic acid (eg, monovalent metal salt of low endotoxin alginic acid) used in the first and second layers is 0.1 mg / cm 2 to 3 mg / mg
  • Any one of the above [2-1] to [2-4], wherein the endotoxin content of the monovalent metal salt of alginic acid in the first layer and the second layer is 500 EU / g or less The adhesion prevention method as described in a term.
  • the monovalent metal salt of alginic acid of the first layer and the second layer is sodium alginate or potassium alginate described in any one of the above [2-1] to [2-5] How to prevent adhesions.
  • the curing agent of the first layer and the second layer is at least one metal ion compound selected from the group consisting of CaCl 2 , CaSO 4 , ZnCl 2 , SrCl 2 , FeCl 3 and BaCl 2
  • the adhesion preventing method according to any one of the above [2-2] to [2-6].
  • the adhesion prevention method as described in.
  • An adhesion prevention method comprising applying a possible sponge-like laminate to a subject in need of adhesion prevention.
  • An adhesion prevention method comprising applying a possible sponge-like laminate to a subject in need of adhesion prevention.
  • the elution amount of the monovalent metal salt of alginic acid in the second layer is 100%.
  • the ratio of the elution amount of the monovalent metal salt of alginic acid in the first layer is less than 50% at 1 hour from the start of measurement and less than 70% at 2 hour, as described above.
  • the first layer contains 25 ⁇ 10% by weight of monovalent metal salt of alginic acid Elution occurs within an hour, 80 ⁇ 10% by weight elutes within 4 hours, and the second layer is an elution of 70 ⁇ 10% by weight of monovalent metal salt of alginic acid within 1 hour, 90 ⁇ 10%.
  • the adhesion preventing method according to any one of the above [2-9] to [2-9b], wherein 10% by weight is eluted within 4 hours.
  • a method for producing an anti-adhesion material including a sponge-like laminate applicable to a living body comprising the following steps: (1) curing the monovalent metal salt of alginic acid having a weight average molecular weight of 10,000 to 2,000,000 with a curing agent, (2) a step of freezing the hardened monovalent metal salt of alginic acid, (3) On the monovalent metal salt of alginic acid obtained in (2), a monovalent metal salt of alginic acid having a weight average molecular weight of 1,000 to 1,000,000 is cured with a curing agent to obtain a laminate.
  • the sponge-like laminate has a sponge-like first layer containing a monovalent metal salt of alginic acid having a weight-average molecular weight of 10,000 to 2,000,000 and a weight-average molecular weight of 1,000 to 1,000,000.
  • a sponge-like second layer containing a monovalent metal salt of alginic acid wherein the weight average molecular weight of the monovalent metal salt of alginic acid in the first layer is higher than that of the second layer, the first layer and the second layer At least one layer selected from layers of at least one pharmaceutical component selected from the group consisting of drugs, growth factors, hormones, proteins and combinations thereof,
  • a sponge-like laminate obtained by incorporating the at least one pharmaceutical ingredient with the monovalent metal salt of alginic acid before curing in step (1) and / or step (3) or after lyophilization in step (4)
  • a method for producing an anti-adhesion material including a sponge-like laminate applicable to a living body comprising the following steps. (1) curing a monovalent metal salt of low endotoxin alginic acid having a weight average molecular weight of 10,000 to 2,000,000 with a curing agent, (2) a step of freezing the hardened monovalent metal salt of alginic acid, (3) On the monovalent metal salt of alginic acid obtained in (2), a low endotoxin alginic acid monovalent metal salt having a weight average molecular weight of 1,000 to 1,000,000 is cured with a curing agent to form a laminate The process of obtaining (4) a step of freeze-drying the obtained laminate to obtain a sponge-like laminate, Here, the molecular weight is measured by GPC-MALS method,
  • the sponge-like laminate comprises a sponge-like first layer containing a monovalent metal salt of low endotoxin alginic acid having a weight average molecular weight of 10,000 to 2,000,000, and
  • a sponge-like laminate applicable to a living body obtained by the following steps (1) to (4): (1) curing the monovalent metal salt of alginic acid having a weight average molecular weight of 10,000 to 2,000,000 with a curing agent, (2) a step of freezing the hardened monovalent metal salt of alginic acid, (3) On the monovalent metal salt of alginic acid obtained in (2), a monovalent metal salt of alginic acid having a weight average molecular weight of 1,000 to 1,000,000 is cured with a curing agent to obtain a laminate.
  • the sponge-like laminate has a sponge-like first layer containing a monovalent metal salt of alginic acid having a weight-average molecular weight of 10,000 to 2,000,000 and a weight-average molecular weight of 1,000 to 1,000,000.
  • a sponge-like second layer containing a monovalent metal salt of alginic acid wherein the weight average molecular weight of the monovalent metal salt of alginic acid in the first layer is higher than that of the second layer, the first layer and the second layer At least one layer selected from layers of at least one pharmaceutical component selected from the group consisting of drugs, growth factors, hormones, proteins and combinations thereof,
  • a sponge-like laminate obtained by incorporating the at least one pharmaceutical ingredient with the monovalent metal salt of alginic acid before curing in step (1) and / or step (3) or after lyophilization in step (4)
  • a sponge-like laminate applicable to a living body obtained by the following steps (1) to (4).
  • the molecular weight is measured by GPC-MALS method
  • the sponge-like laminate comprises a sponge-like first layer containing a monovalent metal salt of low endotoxin alginic acid having a weight average molecular weight of 10,000 to 2,000,000, and a weight average molecular weight of 1,000 to 1,000 to
  • a combination of raw materials for producing an anti-adhesion material comprising at least one pharmaceutical component selected from the group consisting of a factor, a hormone, a protein and a combination thereof.
  • [5-1a] A combination of raw materials according to the above [5-1], wherein the monovalent metal salt of alginic acid is a monovalent metal salt of low endotoxin alginic acid.
  • [5-1b] A first raw material containing monovalent metal salt of low endotoxin alginic acid having a weight average molecular weight of 10,000 to 2,000,000, and low endotoxin alginic acid having a weight average molecular weight of 1,000 to 1,000,000.
  • the first raw material has a weight average molecular weight higher than that of the second raw material, and at least one layer selected from the first layer and the second layer contains A combination of raw materials for producing an anti-adhesion agent, comprising at least one pharmaceutical ingredient selected from the group consisting of drugs, growth factors, hormones, proteins and combinations thereof.
  • the weight average molecular weight is 10,000 to 2,000,000
  • the weight average molecular weight of the monovalent metal salt of alginic acid of the second layer is 1,000 to 1,000,000
  • the weight average molecular weight is It is measured by GPC-MALS after the crosslinking treatment
  • the weight average molecular weight of the monovalent metal salt of alginic acid in the first layer is higher than the weight average molecular weight of the monovalent metal salt of alginic acid in the second layer
  • At least one layer selected from the first layer and the second layer contains at least one pharmaceutical component selected from the group consisting of drugs, growth factors, hormones, proteins and combinations thereof, and is sterilized
  • Living A pharmaceutical composition having an adhesion preventing effect, comprising a sponge-like laminate applicable to the body.
  • the weight average molecular weight of the salt is 10,000 to 2,000,000
  • the weight average molecular weight of the monovalent metal salt of alginic acid in the second layer is 1,000 to 1,000,000
  • the weight average molecular weight Is determined by the GPC-MALS method after the decrosslinking treatment and the weight average molecular weight of the monovalent metal salt of alginic acid in the first layer is greater than the weight average molecular weight of the monovalent metal salt of alginic acid in the second layer.
  • the at least one layer selected from the first layer and the second layer comprises at least one pharmaceutical component selected from the group consisting of drugs, growth factors, hormones, proteins and combinations thereof.
  • a pharmaceutical composition having adhesion-preventing effect comprising a sterile, bio-compatible sponge-like laminate.
  • the total amount of the monovalent metal salt of alginic acid (eg, monovalent metal salt of low endotoxin alginic acid) in the first and second layers is 0.1 mg / cm 2 to 3 mg / mg
  • the monovalent metal salt of alginic acid of the first layer and the second layer is sodium alginate or potassium alginate as described in any one of the above [6-1] to [6-5]
  • Pharmaceutical composition. [6-7] The medicament according to any one of the above [6-1] to [6-6], wherein the curing agent for the first layer and the second layer is a divalent or higher metal ion compound. Composition. [6-7a]
  • the curing agent for the first layer and the second layer is at least one metal ion compound selected from the group consisting of CaCl 2 , CaSO 4 , ZnCl 2 , SrCl 2 , FeCl 3 and BaCl 2.
  • the pharmaceutical composition according to any one of the above [6-1] to [6-6].
  • [6-8] The pharmaceutical composition according to any one of the above [6-1] to [6-7a] for applying the first layer to the wound side surface.
  • [6-9] The pharmaceutical composition according to any one of the above [6-1] to [6-8], wherein the sponge-like laminate is pressed.
  • adhesion-preventing effect is high, both adhesion in wound area and de novo adhesion can be suppressed, no adverse effect on applied living body, healing in wound area not disturbed, intestinal anastomosis etc. It is possible to provide an anti-adhesion material having at least one performance that can be used, is easy to apply via a trocker in endoscopic surgery, is capable of adjusting a sticking position and sticking again, and the like.
  • FIG. 5 shows the evaluation of adhesion formation in a partial liver resection model.
  • A Number of individuals which formed adhesions in separation section,
  • B separation section grade,
  • C separation section Extent (mm). ** p ⁇ 0.01, * p ⁇ 0.05.
  • FIG. 5 shows the evaluation of adhesion formation in a partial liver resection model.
  • A Number of individuals forming adhesions in non-separating section,
  • B Non-separating section grade,
  • FIG. 5 shows the evaluation of adhesion formation in a partial liver resection model.
  • A Number of individuals forming adhesions in non-separating section,
  • B Non-separating section grade,
  • C non-separating section Extent (mm). ** p ⁇ 0.01, * p ⁇ 0.05.
  • FIG. 5 shows the evaluation of adhesion formation in a partial liver resection model.
  • A Number of individuals forming adhesion
  • FIG. 5 shows the evaluation of weight change and spleen weight in a partial liver resection model.
  • A Weight change
  • B Spleen weight
  • FIG. 5 shows the evaluation of adhesion formation in the Pean liver resection model.
  • A Number of individuals which formed adhesions in separation section,
  • B separation section grade
  • C separation section Extent
  • FIG. 5 shows the evaluation of adhesion formation in the Pean liver resection model.
  • A Number of individuals forming adhesions in non-separating section,
  • B Non-separating section grade
  • C non-separating section Extent (mm). ** p ⁇ 0.01, * p ⁇ 0.05.
  • FIG. 5 shows the evaluation of weight change and spleen weight in a partial liver resection model.
  • A Weight change
  • B Spleen weight
  • FIG. 5 shows the evaluation of adhesion formation in the Pean liver resection model.
  • A Number of individuals which formed adhesions in
  • FIG. 5 shows the evaluation of weight change and spleen weight in the Pean liver resection model.
  • A Weight change
  • B Spleen weight. It is a figure which shows the swelling test result of the sponge before and behind a press. It is a figure which shows a time-dependent change of the height (A) of each test-piece tip after spraying, and the angle (B) from a test stand.
  • FIG. 5 is a diagram showing the evaluation of adhesion formation in a cross section in a rat 70% liver resection model.
  • A Number of individuals which formed adhesions in separation section
  • B separation section Extent (mm). ** p ⁇ 0.01, * p ⁇ 0.05.
  • Adhesion Prevention refers to a state in which the surfaces of tissues that should be separated from one another are connected or fused with fibrous tissue.
  • the causes of adhesions include trauma that can be caused on the surface of tissue in surgery, inflammation caused by trauma, and inflammation due to drying of the tissue surface in surgery. With these injuries and inflammation, an exudate containing fibrin is produced on the surface of the tissue, and the exudate is organized to connect or fuse the tissue surfaces to form adhesions.
  • Adhesion prevention refers to reducing the formation of adhesions.
  • the adhesion prevention does not necessarily necessarily completely prevent the formation of adhesions, as long as adhesion formation can be prevented as compared with the state when the adhesion preventive is not applied. That is, “adhesion prevention” may be reworded as alleviation of adhesions, for example, at least one selected from the frequency, range and degree of adhesions should be reduced.
  • the grade of average adhesions is lower than the grade of average adhesions when the anti-adhesion agent is not applied. It should just be.
  • adhesion prevention refers to, for example, the extent evaluation of adhesions described in the examples, the extent of average adhesions being larger compared to the average adhesion extent when no adhesion preventing material is applied. It should be low. "Adhesion prevention” is preferably the prevention of adhesions arising from surgery, more preferably peritoneal adhesions resulting from surgery. That is, “adhesion prevention” is preferably post-surgical adhesion prevention.
  • adhesions to be targeted include adhesions at the excision site of the target organ at the time of surgery and de novo adhesions (adhesion with peripheral areas other than the operation site and with the abdominal site and a wide site in the body) There is.
  • Adhesion prevention material comprises a sponge-like first layer and a second layer of a monovalent metal salt of alginic acid (eg, a monovalent metal salt of low endotoxin alginic acid) at least partially crosslinked with a curing agent
  • a spongy first layer comprising a monovalent metal salt of alginic acid having a relatively high weight average molecular weight (eg, a monovalent metal salt of low endotoxin alginic acid), and a monovalent metal salt of alginic acid having a relatively low weight average molecular weight
  • An anti-adhesion agent hereinafter referred to as “adhesion-preventing agent”
  • one layer comprises at least one pharmaceutical component selected from the group consisting of drugs, growth factors, hormone
  • the weight average molecular weight of the monovalent metal salt of alginic acid (eg, monovalent metal salt of low endotoxin alginic acid) used in the first layer and the second layer is, for example, 10,000 to 2,000,000, respectively It is 1,000 to 1,000,000.
  • Such weight average molecular weight is determined by a decrosslinking treatment, for example, by dissolving in a chelating agent solution and then measuring by GPC-MALS method.
  • the symbol “to” when the symbol “to” is used as the numerical range, it means “more than the lower limit value and less than the upper limit”, and the numerical values at both ends of the symbol are included in the range.
  • the adhesion preventing material A contains monovalent metal salts of alginic acid having different molecular weights (eg, monovalent metal salts of low endotoxin alginic acid) in the first layer and the second layer of the sponge-like laminate. Specifically, the weight average molecular weight of the monovalent metal salt of alginic acid of the first layer is higher than that of the second layer. In the layer containing a monovalent metal salt of alginic acid, the dissolution rate becomes slower as the weight average molecular weight of the monovalent metal salt of alginic acid is larger, while the dissolution rate becomes faster as the weight average molecular weight is smaller.
  • monovalent metal salts of alginic acid having different molecular weights eg, monovalent metal salts of low endotoxin alginic acid
  • the antiadhesive material A with the first layer facing the wound side and the second layer facing the abdominal side, the first layer remains in the wound and the second layer It can be expected that it dissolves relatively quickly and suppresses adhesions in the abdominal cavity.
  • the present invention is also directed to a living organism comprising a first layer and a second layer each comprising a monovalent metal salt of alginic acid (eg, a monovalent metal salt of low endotoxin alginic acid) at least partially crosslinked with a curing agent (eg, a monovalent metal salt of low endotoxin alginic acid).
  • a curing agent eg, a monovalent metal salt of low endotoxin alginic acid.
  • At least one layer comprising an applicable sponge-like laminate, wherein the dissolution rate of the first layer and the second layer is different, and at least one layer selected from the first layer and the second layer is a drug, a growth factor, a hormone
  • An adhesion prevention material (“adhesion prevention material B") is provided, which comprises at least one pharmaceutical component selected from the group consisting of proteins, and combinations thereof.
  • the dissolution rate of the first layer is slower than that of the second layer.
  • the weight average molecular weight of the monovalent metal salt of alginic acid of the first layer as in the antiadhesive material A The degree of crosslinking of the monovalent metal salt of alginic acid in the first layer by making the second layer higher, changing the type of crosslinking agent, or changing the concentration of the crosslinking agent, etc. Second layer And higher.
  • adhesion prevention material B is the first one when the elution amount of the monovalent metal salt of alginic acid in the second layer is 100% in the dissolution test using the elution of the monovalent metal salt of alginic acid as an indicator.
  • the ratio of the elution amount of the monovalent metal salt of alginic acid in the layer is less than 50% at 1 hour from the start of measurement and less than 70% at 2 hour.
  • the adhesion preventing material B is a dissolution test using dissolution of a monovalent metal salt of alginic acid as an indicator
  • the first layer is 25 ⁇ 10% by weight of monovalent metal salt of alginic acid within 1 hour And 80 ⁇ 10% by weight in 4 hours
  • the second layer is 70 ⁇ 10% by weight of monovalent metal salt of alginic acid in 1 hour, 90 ⁇ 10% by weight. % Elutes within 4 hours.
  • the dissolution test is specifically as described in the examples below.
  • anti-adhesion agent B contains 10 to 70 of the pharmaceutical component contained in the first layer in the dissolution test using dissolution of the pharmaceutical component in phosphate buffer at pH 7.5 at 37 ° C. as an indicator. % Dissolves within 1 hour, and 20 to 100% dissolves within 4 hours, more preferably 40 ⁇ 20% by weight of the pharmaceutical component contained in the first layer in the same test Elute within 1 hour and 80 ⁇ 20% by weight elute within 4 hours. In another preferred embodiment, anti-adhesion agent B contains 40 to 100 of the pharmaceutical component contained in the second layer in the dissolution test using dissolution of the pharmaceutical component in phosphate buffer at pH 7.5 and 37 ° C. as an indicator.
  • anti-adhesion agent B contains 10 to 70 of the pharmaceutical component contained in the first layer in the dissolution test using dissolution of the pharmaceutical component in phosphate buffer at pH 7.5 at 37 ° C. as an indicator.
  • % By weight dissolves within 1 hour, 20 to 100% by weight dissolves within 4 hours, and 40 to 100% by weight of the pharmaceutical component contained in the second layer dissolves within 1 hour, 60 to 100% by weight elutes within 4 hours, more preferably 40 ⁇ 20% by weight of the drug component contained in the first layer elutes within 1 hour in the same test, 20% by weight elutes within 4 hours, 80 ⁇ 20% by weight of the drug component contained in the second layer elutes within 1 hour, 90 ⁇ 10% by weight elutes within 4 hours It is a thing.
  • first layer is a layer which is a lower layer when the sponge-like laminate is applied to a target, that is, a layer which is in contact with the surface of the target tissue to be applied.
  • second layer is a layer which becomes an upper layer when the sponge-like laminate is applied to a target, that is, a layer not in contact with the surface of the target tissue to be applied.
  • Bio-applicable means capable of being disposed on the surface of the tissue to be applied as a medical material.
  • the sponge-like laminate applicable to a living body used for the adhesion preventing material may have a third layer containing an optional component other than the first layer and the second layer described above, and And may have a multilayer structure. Also included are sponge-like laminates having a structure in which each layer does not have a clear interface and the molecular weight is continuously increasing or decreasing.
  • the adhesion prevention material 1 includes a sponge-like laminate 4 including a first layer 2 and a second layer 3.
  • the first layer 2 and the second layer 3 are each sponge-like.
  • "Sponge-like" means in a porous state.
  • at least one layer selected from the first layer 2 and the second layer 3 contains at least one pharmaceutical component selected from the group consisting of a drug, a growth factor, a hormone, a protein and a combination thereof .
  • the shape of the sponge-like laminate applicable to a living body is not particularly limited, and can be appropriately selected in consideration of the range of the surface to be applied, the shape, unevenness, and the like.
  • the shape of the sponge-like laminate may be, for example, a flat plate as shown in FIG. 1, or may be a disk, a cylinder, a rectangular parallelepiped, or the like. Preferably it is flat form or disk shape.
  • the adhesion preventing material can be further cut and applied to the surface according to the range, shape, unevenness, etc. of the surface to be applied, the size of the flat plate or disc is not particularly limited.
  • the length and width are not particularly limited, and the height (thickness) is preferably 0.2 mm to 30 mm. More preferably, it is 0.3 mm to 15 mm, more preferably 0.5 mm to 10 mm. More preferably, in addition to such height (thickness), the length and width are 1 mm to 300 mm ⁇ 1 mm to 300 mm, and particularly preferably 3 mm to 200 mm ⁇ 3 mm to 200 mm. More preferably, it is 5 mm to 150 mm ⁇ 5 mm to 150 mm.
  • the thickness does not have to be uniform, and one may be thick and the other may be thin and inclined.
  • the sponge-like laminate of the anti-adhesion material of the preferred embodiment has high flexibility and is less likely to be broken as compared with Seprafilm (trade name).
  • the sponge-like laminate is pressed. "Press" is as described later.
  • its height (thickness) is preferably 0.02 mm to 3 mm, more preferably 0.03 mm to 1.5 mm, still more preferably 0.05 mm to It is 1 mm.
  • the length and width are 1 mm to 300 mm ⁇ 1 mm to 300 mm, and particularly preferably 3 mm to 200 mm ⁇ 3 mm to 200 mm. More preferably, it is 5 mm to 150 mm ⁇ 5 mm to 150 mm.
  • the thickness after pressing is uniform.
  • At least one pharmaceutical ingredient is included in the first layer but not in the second layer. In another aspect, at least one pharmaceutical ingredient is included in the second layer but not included in the first layer. In yet another aspect, the at least one pharmaceutical ingredient is independently included in both the first layer and the second layer. The at least one pharmaceutical ingredient is, for example, three or more, two or one pharmaceutical ingredient.
  • the drug is a drug having an action as an antibiotic, a drug having an inflammation reducing action, a drug having an adhesion preventing action, a drug having a wound healing promoting action, a drug having a hemostatic action and the like.
  • Such drugs are not limited to, for example: Penicillin antibiotics such as penicillin, ampicillin, amoxicillin, cephem antibiotics such as cefazolin, cefotiam, ceftazim, cefpirome, carbapenem antibiotics such as meropenem, monobactam antibiotics such as tazobactam, penem antibiotics such as faropenem Aminoglycoside antibiotics such as streptomycin, tobramycin, amikacin, gentamycin, neomycin, lincomycin antibiotics such as lincomycin, fosfomycin antibiotics such as fosfomycin, tetracycline antibiotics such as tetracycline and minocycline, chloramphenicol etc.
  • Penicillin antibiotics such as penicillin, ampicillin, amoxicillin
  • cephem antibiotics such as cefazolin, cefotiam, ceftazim, cefpirome
  • carbapenem antibiotics such as
  • Macrolide antibiotics such as chloramphenicol antibiotics, erythromycin, clarithromycin, azithromycin and josamycin
  • Ketolide antibiotics such as telithromycin, polypeptide antibiotics such as polymyxin, glycopeptide antibiotics such as vancomycin, streptogramin antibiotics such as quinupristin and dalhopristin, nalidixic acid, enoxacin, levofloxacin, gatifloxacin etc Quinolone antibiotics, oxazolidinone antibiotics such as linezolid, and antibiotics having antibacterial and antifungal activity such as amphotericin B, Non-steroidal anti-inflammatory drugs (NSAIDs, for example, aspirin, loxoprofen, diclofenac sodium, indomethacin, piroxicam, mefenamic acid, sulpyrin, acetaminophen, ibuprofen, meloxicam, chlorpheniramine maleate, etc.),
  • the drug is a nonsteroidal antiinflammatory drug or a steroidal antiinflammatory drug, more preferably a steroidal antiinflammatory drug (eg, dexamethasone, dexamethasone sodium phosphate ester, triamcinolone acetonide, etc.).
  • a steroidal antiinflammatory drug eg, dexamethasone, dexamethasone sodium phosphate ester, triamcinolone acetonide, etc.
  • these drugs may be obtained commercially or may be produced by known methods.
  • growth factors are used to promote wound healing.
  • growth factors include, for example, bone morphogenetic protein (BMP), fibroblast growth factor (FGF), vascular endothelial growth factor (VEGF), hepatocyte growth factor (HGF), transforming growth factor (TGF) Insulin-like growth factor (IGF), platelet derived growth factor (PDGF), CDMP (cartilage-derived-morphogenetic protein), colony stimulating factor (CSF), erythropoietin (EPO), thrombopoietin (TPO), interleukin (IL), Platelet-rich plasma (PRP (Platelet Rich Plasma)), SOX, IF, epidermal growth factor (EGF), nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), NT3 and the like, preferably FGF, VE F, NGF, IGF, PDGF and HGF and the like, more preferably FGF and VEGF.
  • BMP bone morphogenetic protein
  • FGF fibroblast growth factor
  • hormones are used to alleviate inflammation, prevent adhesions, promote wound healing and the like.
  • Such hormones are, for example, anabolic hormones such as stanozolol, growth hormone, leptin and the like, preferably growth hormone and leptin. These hormones may be obtained commercially or may be produced by known methods.
  • proteins also include peptides having physiological activity, and are used for inflammation reduction, adhesion prevention, wound healing promotion, hemostasis, pain alleviation and the like.
  • proteins are, for example, thrombin, plasmin, urinastatin, anti-TNF antibody, anti-NGF antibody, TSG-6 and hemocoagulase etc., preferably thrombin and urinastatin.
  • proteins may be obtained commercially or may be produced by known methods.
  • combinations of drugs, growth factors, hormones and proteins include, for example, the following combinations.
  • Non-steroidal anti-inflammatory drug and growth factor combination non-steroidal anti-inflammatory drug and protein combination
  • steroidal anti-inflammatory drug and growth factor combination steroidal anti-inflammatory drug and protein combination
  • steroidal anti-inflammatory drug and protein combination etc.
  • a combination of diclofenac sodium and FGF a combination of diclofenac sodium and thrombin, a combination of diclofenac sodium and ulinastatin, a combination of dexamethasone sodium phosphate and FGF, dexamethasone sodium phosphate and Examples include, but are not limited to, combinations of thrombin, combinations of sodium dexamethasone phosphate and urinastatin, and the like.
  • Preferred pharmaceutical ingredients to be included in the first layer are: Pharmaceutical components and antibiotics having a hemostatic action for wound healing, antibiotics, specifically, any hemostatic agent, any hormone, thrombin, plasmin, ulinastatin and the like, more specifically tranexamic acid, FGF, growth Hormone, thrombin and ulinastatin etc.
  • another preferable pharmaceutical ingredient to be contained in the first layer includes a lipid-soluble pharmaceutical ingredient.
  • another preferable pharmaceutical ingredient to be contained in the first layer includes a pharmaceutical ingredient to be contained for sustained release.
  • Preferred pharmaceutical ingredients to be included in the second layer are: Pharmaceutical components and antibiotics having anti-inflammatory and anti-adhesion effects, specifically, non-steroidal anti-inflammatory drugs, steroid anti-inflammatory drugs, anti-TNF antibodies, anti-NGF antibodies, TSG-6, etc. Specifically, diclofenac sodium, dexamethasone sodium phosphate ester and the like can be mentioned.
  • another preferable pharmaceutical ingredient to be contained in the second layer includes a water-soluble pharmaceutical ingredient.
  • another preferable pharmaceutical ingredient to be contained in the second layer is a pharmaceutical ingredient to be contained for immediate release after surgery.
  • the pharmaceutical component of the first layer and the pharmaceutical component of the second layer are each independently selected. That is, the same drug component may be contained in the first layer and the second layer, or different drug components may be contained in the first layer and the second layer.
  • first and second layers contain the same pharmaceutical component, at least one (for example, three or more, two or one) pharmaceutical components included in the first layer, and the second The at least one (eg, three or more, two or one) pharmaceutical ingredients contained in the layer of.
  • the first layer contains one pharmaceutical component and the second layer also contains the same one pharmaceutical component.
  • Preferred pharmaceutical ingredients when the first and second layers contain the same pharmaceutical ingredient are, for example, antibiotics.
  • the first layer and the second layer contain different pharmaceutical ingredients, at least one (for example, three or more, two or one) pharmaceutical ingredients contained in the first layer, and the second layer And at least one (eg, three or more, two or one) of the pharmaceutical ingredients do not completely match. That is, the inclusion of different medicinal components in the first layer and the second layer means that there is no overlap in at least one medicinal component of the first layer and at least one medicinal component of the second layer, And the at least one pharmaceutical component of the first layer and the at least one pharmaceutical component of the second layer are overlapping, but the overlapping is only a part.
  • the first layer contains one pharmaceutical ingredient and the second layer contains another pharmaceutical ingredient.
  • a preferred combination of pharmaceutical ingredients in the case of containing different pharmaceutical ingredients in the first layer and the second layer is the following combination.
  • First layer one or more of any medicinal component having a wound healing promoting action, hemostatic action, and the second layer: one or more of any pharmaceutical component having a inflammation reducing action, an adhesion preventing action Combination of
  • First layer one or more of any medicinal component having a wound healing promoting action, hemostatic action, and the second layer: one selected from non-steroidal anti-inflammatory drug, steroidal anti-inflammatory drug Combination with the above.
  • First layer A combination of one or more selected from FGF, thrombin, ulinastatin and a second layer: one or more selected from diclofenac sodium, dexamethasone sodium phosphate ester.
  • the anti-adhesion agent of some embodiments can be used as a pharmaceutical composition by containing a pharmaceutical component as described above.
  • the adhesion preventing material of the preferred embodiment enhances the effect of the pharmaceutical component by containing and applying the pharmaceutical component, that the effect can be obtained in an amount smaller than the clinical dose of the pharmaceutical component, and the pharmaceutical component At least one of the following effects may be obtained: reducing adverse events, continuously releasing the pharmaceutical component, retaining the pharmaceutical component locally, and controlling the release rate of the pharmaceutical component. is there.
  • the content of the pharmaceutical component to be contained in the adhesion prevention material of some embodiments is not particularly limited as long as the effect of the pharmaceutical component is exhibited and does not show adverse events, and the efficacy of the pharmaceutical component to be contained
  • the sponge-like laminate is porous and has a water absorbability, for example, compared with the nonporous Seprafilm (trade name), it is easy to contain the pharmaceutical component by preparation at the time of use. For example, by impregnating a solution containing a pharmaceutical ingredient with a sponge and administering it, adhesion prevention and local sustained release of the pharmaceutical ingredient can be achieved simultaneously in the abdominal cavity, chest cavity, heart cavity and the like. Furthermore, the inclusion of the pharmaceutical component in layers having different dissolution rates also enables the sustained release of the pharmaceutical component at a high sustained release rate and a slow sustained release rate.
  • the sponge-like laminate exhibits an excellent adhesion prevention effect by containing the pharmaceutical component as compared with the one without the pharmaceutical component.
  • the present invention also includes a sponge-like first layer and a second layer of a monovalent metal salt of alginic acid (eg, a monovalent metal salt of low endotoxin alginic acid) at least partially crosslinked with a curing agent,
  • a sponge-like first layer containing a monovalent metal salt of alginic acid having a relatively high weight average molecular weight eg, a monovalent metal salt of low endotoxin alginic acid
  • a monovalent metal salt of alginic acid having a relatively low weight For example, at least one member selected from a first layer and a second layer, comprising a sponge-like sponge-like laminate including a sponge-like second layer containing a monovalent metal salt of low endotoxin alginic acid
  • a pharmaceutical composition is provided, wherein the layer comprises at least one pharmaceutical component selected from the group consisting of drugs, growth factors, hormones, proteins and combinations thereof.
  • the weight average molecular weight of the monovalent metal salt of alginic acid (eg, monovalent metal salt of low endotoxin alginic acid) used in the first layer and the second layer is, for example, 10,000 to 2,000,000, respectively It is 1,000 to 1,000,000.
  • Such weight average molecular weight is determined by a decrosslinking treatment, for example, by dissolving in a chelating agent solution and then measuring by GPC-MALS method.
  • the description about the sponge-like laminate, the pharmaceutical component and the like is the same as above.
  • Alginate Monovalent Metal Salt is a water-soluble salt formed by ion-exchange of the hydrogen atom of the 6-position carboxylic acid of alginic acid with a monovalent metal ion such as Na + or K + It is salt.
  • a monovalent metal ion such as Na + or K + It is salt.
  • Specific examples of the monovalent metal salt of alginic acid include sodium alginate, potassium alginate and the like, and in particular, sodium alginate which is commercially available is preferable.
  • the solution of monovalent metal salt of alginic acid forms a gel when mixed with the curing agent.
  • alginic acid is a biodegradable high molecular weight polysaccharide, which is a polymer obtained by linear polymerization of two types of uronic acid, D-mannuronic acid (M) and L-guluronic acid (G). is there. More specifically, the homopolymer fraction of D-mannuronic acid (MM fraction), the homopolymer fraction of L-guluronic acid (GG fraction), and the arrangement of D-mannuronic acid and L-guluronic acid at random These fractions (MG fraction) are block copolymers optionally combined.
  • the compositional ratio (M / G ratio) of D-mannuronic acid to L-guluronic acid of alginic acid differs mainly depending on the type of organism from which seaweed etc. is derived, and is influenced by the growing place and season of the organism,
  • the high G-type having a M / G ratio of about 0.4 to the high M-type having a M / G ratio of about 5 covers a high range.
  • the monovalent metal salt of alginic acid is a high molecular weight polysaccharide, and it is difficult to determine the molecular weight accurately, and it is known that differences in values may occur depending on the measurement method when measuring the molecular weight of high molecular weight substances derived from natural products. ing.
  • the absolute weight average molecular weight can be measured.
  • the weight average molecular weight of the monovalent metal salt of alginic acid used as a raw material measured by GPC-MALS method is, for example, 10,000 to 2,000,000 in the first layer of the sponge-like laminate, and preferably 15,000 to 1,500,000, more preferably 20,000 to 1,000,000, and particularly preferably 25,000 to 500,000.
  • the weight average molecular weight measured by the GPC-MALS method is, for example, 1,000 to 1,000,000 in the second layer, preferably 1 And more preferably 2,000 to 250,000, and particularly preferably 3,000 to 100,000.
  • the weight average molecular weight measured by the GPC-MALS method is, for example, 10,000 to 10,000 in the first layer of the sponge-like laminate. It is 300,000, preferably 10,000 to 200,000, more preferably 10,000 to 100,000, and particularly preferably 10,000 to 80,000.
  • the weight average molecular weight measured by the GPC-MALS method is, for example, 1,000 to 100,000 in the second layer, preferably 1,000. To 80,000, more preferably from 2,000 to 60,000, and particularly preferably from 3,000 to 60,000.
  • the monovalent metal salt of alginic acid crosslinked at least partially with a curing agent is a weight average molecular weight as a monovalent metal salt of non-crosslinked alginic acid by measurement by GPC-MALS method after arbitrary decrosslinking treatment Can be measured.
  • the decrosslinking treatment for example, dissolving in any chelating agent such as a chelating agent solution such as EDTA (ethylenediaminetetraacetic acid), phytic acid and the like can be mentioned.
  • EDTA ethylenediaminetetraacetic acid
  • EDTA ethylenediaminetetraacetic acid
  • the weight average molecular weight of the monovalent metal salt of alginic acid in the first layer of the sponge-like laminate is higher than in the second layer.
  • the weight average molecular weight of the raw material of the sponge-like laminate or the monovalent metal salt of alginic acid of the first layer contained in the sponge-like laminate is, for example, 1,000 to 1,000,000 higher than that of the second layer. , Preferably 2,000 to 500,000, and more preferably 3,000 to 300,000.
  • the molecular weight of the high molecular weight polysaccharide is calculated by the above-mentioned method, a measurement error of 10 to 20% by weight can occur. For example, 8,000 to 12,000 for 10,000, 80,000 to 120,000 for 100,000, and 160,000 to 240,000 for 400,000. In the case of 320,000 to 480,000 and 500,000, fluctuation of values may occur in the range of about 400,000 to 600,000.
  • the molecular weight of alginic acid can be measured according to a conventional method. Typical conditions for using GPC-MALS for molecular weight determination are as described in Example 1 herein.
  • a detector for example, an RI detector and a light scattering detector (MALS) can be used.
  • Alginates generally have a large molecular weight initially when extracted from brown algae, but their molecular weight gradually decreases in the process of drying by heat, purification, and the like. Alginates having different molecular weights can be produced by techniques such as condition control such as temperature in the production process, selection of brown algae as a raw material, and fractionation of molecular weight in the production process. Furthermore, it is also possible to obtain alginic acid having a target molecular weight by mixing with another lot of alginic acid having different molecular weight.
  • the monovalent metal salt of alginic acid used herein is, for example, one treated with low endotoxin.
  • the monovalent metal salt of alginic acid may not be one that has been confirmed to be low endotoxin treatment or may be one that has not been low endotoxin treatment.
  • the low endotoxin treatment can be performed by a known method or a method analogous thereto.
  • the method of the present invention for purifying sodium hyaluronate see, eg, JP-A-9-324001
  • the method of Yoshida et al. For purifying ⁇ 1,3-glucan (eg, JP-A-8-269102).
  • Low endotoxin treatment is not limited to them, but it is used for washing, filtration with filter (endotoxin removal filter, charged filter, etc.), ultrafiltration, column (endotoxin adsorption affinity column, gel filtration column, column with ion exchange resin, etc.) Purification, adsorption to hydrophobic substance, resin or activated carbon, etc., organic solvent treatment (extraction with organic solvent, precipitation / precipitation by addition of organic solvent, etc.), surfactant treatment (see, eg, JP-A-2005-036036) These can be carried out according to known methods such as) or a combination of these as appropriate. Well-known methods such as centrifugation may be appropriately combined with the steps of these treatments. It is desirable to select appropriately according to the type of alginic acid.
  • the endotoxin level can be confirmed by a known method, and can be measured, for example, by a method using Limulus reagent (LAL), a method using Endospecy (registered trademark) ES-24S set (Seikagaku Kogyo Co., Ltd.), etc. .
  • LAL Limulus reagent
  • Endospecy registered trademark
  • ES-24S set Seikagaku Kogyo Co., Ltd.
  • the method for treating endotoxin of monovalent metal salt of alginic acid used here is not particularly limited, but as a result, the endotoxin content of the bioabsorbable polysaccharide is determined when the endotoxin is measured by the Limulus reagent (LAL). It is preferably at most 500 endotoxin units (EU) / g, more preferably at most 100 EU / g, particularly preferably at most 50 EU / g, particularly preferably at most 30 EU / g.
  • Sodium alginate is low endotoxin treatment, for example, Sea Matrix (TM) (Mochida Pharmaceutical Co., Ltd.) are available by commercially such PRONOVA TM UP LVG (FMCBioPolymer).
  • the use amount of the monovalent metal salt of alginic acid in the sponge-like laminate can be appropriately selected in consideration of the adhesion preventing effect.
  • the amount of monovalent metal salt of alginic acid, the sum of the first and second layers of sponge-like laminate for example, 0.1mg / cm 2 ⁇ 10.0mg / cm 2 and the like, preferably It is 0.1 mg / cm 2 to 3.0 mg / cm 2 , more preferably 0.5 mg / cm 2 to 2.5 mg / cm 2 , still more preferably 1.8 mg / cm 2 to 2 mg / cm 2. It is 2 mg / cm 2 , particularly preferably 2.0 mg / cm 2 .
  • the adhesion amount of the monovalent metal salt of alginic acid is 1.0 mg / cm 2 to 3.0 mg / cm 2 in total of the first layer and the second layer of the sponge-like laminate, whereby adhesion is higher. Preventive effect can be expected. If the amount used is 10.0 mg / cm 2 or less, there is little risk of adverse events such as accumulation in the living body and hypertrophy of a specific organ, and if the amount used is 0.1 mg / cm 2 or more, sufficient adhesion preventing effect Can be expected.
  • the ratio (weight ratio) of the used amount of monovalent metal salt of alginic acid in the first layer and the second layer is preferably 1:20 to 20: 1, and more preferably 1: 5 to 5: 1, more preferably 1: 3 to 3: 1, and particularly preferably 1: 2 to 2: 1.
  • one of the first layer and the second layer may contain a curing agent (ie, one of the first layer and the second layer does not contain a curing agent). Or the first layer and the second layer may contain a curing agent. Alternatively, in the anti-adhesion agent, neither the first layer nor the second layer may contain a curing agent.
  • the first layer and the second layer are at least partially crosslinked with a curing agent.
  • the curing agent cures by crosslinking a solution of a monovalent metal salt of alginic acid.
  • the curing agent include metal ion compounds having a valence of 2 or more such as Ca 2+ , Mg 2+ , Ba 2+ , Sr 2+ , Zn 2+ , Fe 3+ and the like, and crosslinkable reagents having 2 to 4 amino groups in the molecule. It can be mentioned.
  • the amount of the curing agent used in the first layer and the second layer is preferably adjusted appropriately in accordance with the amount and molecular weight of the monovalent metal salt of alginic acid.
  • the amount of the curing agent used in the first layer is, for example, 0.1 ⁇ mol / cm 2 to 100 ⁇ mol / cm 2 , preferably 0.5 ⁇ mol / cm 2 to 2.0 ⁇ mol / cm. 2
  • the amount of the curing agent used in the second layer is, for example, 0.1 ⁇ mol / cm 2 to 10 ⁇ mol / cm 2 , preferably 0.6 ⁇ mol / cm 2 to 2.4 ⁇ mol / cm. 2
  • the anti-adhesion material including a bio-applicable sponge-like laminate and the bio-applicable sponge-like laminate can be produced, for example, through the following steps.
  • At least one pharmaceutical component selected from the group consisting of a drug, a growth factor, a hormone, a protein and a combination thereof is used as a monovalent metal of alginic acid before curing in step (1) and / or step (3). It is contained for the salt or for the first and / or second layer of the sponge-like laminate after lyophilization of step (4). Although any method may be used, from the viewpoint of homogeneously containing the pharmaceutical component throughout the sponge, the method of incorporating it to the monovalent metal salt of alginic acid before curing in step (1) and / or step (3) Is more desirable.
  • first alginate a monovalent metal salt of alginic acid having a weight average molecular weight of 10,000 to 2,000,000 (for example, a monovalent metal salt of low endotoxin alginic acid) (hereinafter, “first alginate”)
  • first alginate a monovalent metal salt of low endotoxin alginic acid
  • the solution of the first alginate and the solution of the curing agent can be prepared by a known method or a method analogous thereto.
  • the solvent is not particularly limited as long as it is a solvent applicable to a living body, but is preferably an aqueous solvent, for example, purified water, pure water (for example, distilled water, ion exchanged water), milliQ water, physiological saline, Phosphate buffered saline, DMSO or the like, more preferably pure water. It is preferable that these be sterilized, and those treated with low endotoxin are preferable.
  • an aqueous solvent for example, purified water, pure water (for example, distilled water, ion exchanged water), milliQ water, physiological saline, Phosphate buffered saline, DMSO or the like, more preferably pure water. It is preferable that these be sterilized, and those treated with low endotoxin are preferable.
  • the first alginate can be cured by mixing the solution of the first alginate and the solution of the curing agent.
  • the pharmaceutical component When the pharmaceutical component is contained relative to the monovalent metal salt of alginic acid before curing in step (1), at least one pharmaceutical component is contained relative to the solution of the first alginate, and then the hardening agent is The first alginate is cured by further mixing the solution. Thereby, the hardened first alginate contains at least one pharmaceutical ingredient.
  • the pharmaceutical component to be contained you may make it process microcapsulation, liposome formation, etc. beforehand.
  • the 1st alginate hardened at step (1) is frozen by a conventional method.
  • the mixing ratio of the first layer and the second layer can be reduced.
  • the freezing temperature and time are, for example, 4 hours at -20.degree.
  • a monovalent metal salt of alginic acid having a weight average molecular weight of 1,000 to 1,000,000 for example, a monovalent metal salt of low endotoxin alginic acid
  • second alginate a monovalent metal salt of low endotoxin alginic acid
  • the solution of the second alginate and the solution of the curing agent can be prepared by a known method or a method analogous thereto.
  • the solvent is the same as that described in the above step (1).
  • the solution of the second alginate contains at least one pharmaceutical component, and then the hardening agent is The second alginate is cured by further mixing the solution.
  • the hardened second alginate contains at least one pharmaceutical ingredient.
  • the second alginate can be cured by mixing the solution of the second alginate and the solution of the curing agent.
  • a further hardened second alginate may be frozen prior to step (4).
  • the freezing temperature and time are, for example, 4 hours at -20.degree.
  • the laminate obtained in the step (3) is freeze-dried to obtain a sponge-like laminate.
  • Lyophilization can be carried out by known methods. The conditions of lyophilization can be appropriately adjusted, and a primary drying step, a secondary drying step, and the like may be provided.
  • the pharmaceutical component When the pharmaceutical component is contained in the first layer and / or the second layer of the sponge-like laminate after lyophilization in step (4), for example, a solution containing the pharmaceutical component is used as the first layer and And / or dripping to the second layer to remove the solvent in the solution, spraying a solution containing the pharmaceutical component onto the first layer and / or the second layer, to the solution containing the pharmaceutical component By immersing the layer and / or the second layer, spraying the powder containing the pharmaceutical ingredient onto the first layer and / or the second layer of the sponge-like laminate swollen with saline or the like Do.
  • the pharmaceutical component can be contained in the first layer and / or the second layer of the sponge laminate.
  • the alginic acid of the first layer includes a sponge-like first layer containing a first alginate and a hardening agent, and a sponge-like second layer containing a second alginate and a hardening agent.
  • At least one layer selected from the first layer and the second layer is a drug, a growth factor, a hormone, a protein, and a combination thereof, in which the weight average molecular weight of the monovalent metal salt of A bio-applicable sponge-like laminate comprising at least one pharmaceutical component selected from the group consisting of: and an anti-adhesion material comprising the sponge-like laminate can be obtained.
  • a sponge-like first layer containing a first alginate and a curing agent is prepared, and a sponge-like second layer containing a second alginate and a curing agent is formed thereon.
  • the second layer may be formed, and the first layer may be formed thereon.
  • the sponge-like laminate can be produced, for example, through the following steps. (1 ') a step of curing the second alginate with a curing agent, (2 ') freezing the hardened second alginate; (3 ′) On the second alginate obtained in (2 ′), curing the first alginate with a curing agent to obtain a laminate (4 ') A step of freeze-drying the obtained laminate to obtain a sponge-like laminate.
  • At least one pharmaceutical ingredient selected from the group consisting of a drug, a growth factor, a hormone, a protein, and a combination thereof is used as a second alginate before curing of step (1 ′) and / or step (3) 'For the first alginate before curing, or for the first and / or second layer of the spongy laminate after lyophilization of step (4') .
  • the specific description of each step is the same as the method described above.
  • the method of making a sponge-like laminated body contain a pharmaceutical component is also the same as that of said method.
  • a sponge-like first layer is prepared by curing and lyophilizing the first alginate, and separately a sponge-like second layer by curing and lyophilizing the second alginate. It is also possible to obtain a sponge-like laminate by preparing the above and bonding the obtained sponge-like layers.
  • at least one pharmaceutical component selected from the group consisting of a drug, a growth factor, a hormone, a protein and a combination thereof is used as a first alginate before curing and / or a second alginate before curing. Or the first layer and / or the second layer of the sponge-like laminate after lamination.
  • the specific description of each step is the same as the method described above.
  • the method of making a sponge-like laminated body contain a pharmaceutical component is also the same as that of said method.
  • the desired size can be obtained by using a container, mold, substrate, porous membrane, non-woven fabric, woven fabric, etc. of desired size, height and shape.
  • An anti-adhesion material can be obtained, including sponge-like laminates, height and shape.
  • the sponge-like laminate of the adhesion preventing material is preferably further subjected to sterilization treatment.
  • Sterilization includes, but is not limited to, gamma ray sterilization, electron beam sterilization, ethylene oxide gas sterilization, ethanol sterilization and the like.
  • the anti-adhesion agent is sterilized by electron beam and / or gamma irradiation.
  • a highly biocompatible medical material in which the storage property in the living body is controlled can be obtained (see, for example, JP-A-2000-237294). .
  • the irradiation conditions for electron beam and / or ⁇ -ray sterilization include, for example, absorbed dose of 10 kGy to 150 kGy, more preferably 20 kGy to 100 kGy, and still more preferably 40 kGy to 80 kGy.
  • absorbed dose is 20 kGy to 80 kGy, 20 kGy to 60 kGy, 40 kGy to 60 kGy and the like.
  • Electron beam sterilization is preferred to ⁇ -ray sterilization.
  • the method further includes the step of pressing the laminate obtained in the step (4) or the like.
  • the press is performed by sandwiching and pressing the laminate manually or with a press.
  • generally used processes such as compression and thinning are included in the press herein.
  • the pressing pressure is, for example, 1 kPa to 100 MPa, more preferably 10 kPa to 80 MPa, and still more preferably 100 kPa to 60 Mpa.
  • the manual pressing is performed via a press that can be pressed by hand so that the laminate is uniformly pressed, for example, an acrylic ruler, an acrylic plate, a glass plate, a metal plate or the like.
  • a hot press (AH-1T manufactured by As One Corporation) may be mentioned.
  • the anti-adhesion agent or the pharmaceutical composition (hereinafter referred to as "adhesion-preventing material") is used by applying to a subject in need of anti-adhesion.
  • the adhesion preventing material is absorbed and degraded after staying at the application site, usually for about one week required to exert the adhesion preventing effect, and finally eliminated from metabolism and excretion in about 1 to 2 months. Excellent in safety.
  • the anti-adhesion agent may be applied to the surface of the wound, for example the surface of tissue associated with surgery.
  • tissue related to surgery is a tissue which has been damaged on the surface in the surgery, or a tissue in which the surface is dried in the surgery to cause inflammation or inflammation.
  • Tissues associated with surgery are preferably organs that are enclosed in the peritoneum (eg, stomach, jejunum, ileum, appendix, colon, liver, spleen, duodenum, and pancreas).
  • the anti-adhesion material of a preferred embodiment can effectively prevent serious adhesions such as adhesions that occur after hepatectomy.
  • applying refers to placing the anti-adhesion agent on the surface of the wound (eg, the surface of the tissue associated with the surgical procedure).
  • the surface of the first layer of the sponge-like laminate is in contact with the surface on the wound side (e.g., the surface of the tissue), and the surface of the second layer is the surface on the wound side (e.g.
  • the anti-adhesion agent is placed on the wound-side surface (eg, the surface of the tissue associated with the surgical procedure) so as to face the opposite side (eg, the abdominal cavity side) to the surface).
  • the first layer of the sponge-like laminate has a relatively high weight average molecular weight, so it remains unresolved for a sufficient time to prevent adhesions on the tissue surface and acts as a physical barrier on the wound surface.
  • the second layer of the sponge-like laminate has a relatively low weight-average molecular weight, it rapidly melts and spreads, and plays a role in preventing adhesion on the non-wound surface.
  • the sponge-like laminate of the anti-adhesion material has high flexibility and is less likely to be broken as compared with Seprafilm (trade name).
  • the anti-adhesion agent is not defined on the surface of the tissue to which it is applied, and can be used by, for example, winding it around the intestinal tract at the time of intestinal anastomosis.
  • the surgical instrument can be easily inserted from the passage to and from the subject during endoscopic surgery.
  • the anti-adhesion material can be re-sticked.
  • the sponge laminate of the anti-adhesion material has a wide range of anti-adhesion targets as compared with INTERCEED (trade name).
  • the anti-adhesion agent is prepared in an appropriate size depending on the area, shape, unevenness, etc. of the surface to be applied, and is applied to the surface of the tissue associated with the surgery to be anti-adhesion.
  • Subject is a human or non-human organism, eg, birds and non-human mammals (eg, cattle, monkeys, cats, mice, rats, guinea pigs, hamsters, pigs, dogs, dogs, rabbits, sheep, and horses) is there.
  • the sponge laminate of the adhesion preventing material particularly the sponge laminate
  • it can be compactly assembled, so that, for example, the adhesion preventing material is relatively attached to the affected area through the trocar etc. in endoscopic surgery. It can be applied easily.
  • the adhesion preventing material applied to the affected area preferably absorbs the moisture present in the affected area or the fluid applied to the affected area to recover its thickness.
  • the anti-adhesion agent can be used safely in the subject, as is the case with Seprafilm (trade name) or INTERCEED (trade name).
  • tissue associated with the surgery Although it is not usually necessary to apply anti-adhesion material to the surface of the tissue associated with the surgery after application to the surface of the tissue associated with the surgery, it is necessary, if necessary, for the tissue associated with the anti-adhesive material and surgery.
  • the surface may be sutured.
  • the specific method is as described above.
  • a sponge-like laminate for adhesion prevention is provided.
  • a specific sponge-like laminate is as described above.
  • a kit comprising a sponge-like laminate and a pharmaceutical component for preparing an adhesion preventive material or pharmaceutical composition containing a pharmaceutical component at the time of use, and an adhesion preventive material or pharmaceutical composition comprising a pharmaceutical component at the time of use
  • a method is provided for using the sponge-like laminate and the pharmaceutical component in combination.
  • Specific sponge-like laminates and pharmaceutical ingredients are as described above.
  • non-steroidal anti-inflammatory drugs can be used as anti-adhesion agents before, simultaneously with, or after application of anti-adhesion agents or pharmaceutical compositions to tissue associated with surgery.
  • Combination drugs such as NSAIDs
  • steroidal anti-inflammatory drugs may be administered separately from the adhesion prevention material.
  • Example 1 Preparation of Alginate Laminated Sponge An alginate laminated sponge was prepared as follows.
  • stacking sponge is as follows.
  • Low endotoxin sodium alginate was obtained from Mochida Pharmaceutical Co., Ltd.
  • AL10 (Lot NO. 5K12202), endotoxin amount 4 EU / g.
  • Calcium chloride was obtained from Wako Pure Chemical Industries, Ltd. (product code: 036-00485).
  • Preparation procedure (1) Preparation of Solution AL 500 was dissolved in pure water at a concentration of 1.0 wt% to prepare an AL 500 solution. Similarly, AL10 was dissolved in pure water at a concentration of 1.0 wt% to prepare an AL10 solution. Furthermore, calcium chloride was dissolved in pure water to prepare 10 mM and 15 mM aqueous solutions of calcium chloride, respectively.
  • An alginate laminate sponge of interest comprises a spongy lower layer (ie, a first layer) comprising AL 500 and calcium chloride, and a sponge-like upper layer (ie, a second layer) comprising AL 10 and calcium chloride.
  • the total amount of sodium alginate used in the upper and lower layers was about 2.0 mg / cm 2 .
  • the ratio (weight ratio) of the amount used of the upper and lower layers of sodium alginate was 1: 1.
  • the amount of calcium chloride, the upper layer is about 1.0 [mu] mol / cm 2, at the lower, it was about 1.5 ⁇ mol / cm 2.
  • Pre-processing method The sample was dissolved by adding an eluent to the sample, and filtered through a 0.45 ⁇ m membrane filter to obtain a measurement solution.
  • a single layer sponge containing AL10 manufactured according to the method of the above steps (1), (2) and (4) and a single layer sponge containing AL 500 are subjected to electron beam sterilization, and then EDTA (ethylenediaminetetraacetic acid) It was dissolved in a solution, and the molecular weight was measured by GPC-MALS method. The results are shown below.
  • Example 3 and Examples 3-2 and 4 described later the laminated sponge obtained was subjected to electron beam sterilization (20 kGy) as the laminated sponge.
  • Example 6 and 7 described later non-sterile ones were used as the laminated sponges.
  • Example 1-2 Preparation of Alginate Laminated Sponge (1) Preparation of Sponge Using alginic acid described in the following [Reagent], using AL100 or AL500 as the lower layer material and using AL10 or AL20 as the upper layer material Each alginic acid laminated sponge was prepared according to the method described in Example 1 by combining AL10 (upper layer)-AL 100 (lower layer), AL 20 (upper layer)-AL 100 (lower layer), AL 20 (upper layer)-AL 500 (lower layer). .
  • Example 2 Measurement of Dissolution Rate of Each Layer of Alginate Laminated Sponge
  • a layered sponge in which the upper layer or the lower layer was fluorescently modified was prepared, and the dissolution rate was measured.
  • the specific method is shown below.
  • FTSC Fluorescein-5-Thiosemicarbazide
  • the labeling of alginic acid was performed by a conventional method.
  • a laminated sponge was produced according to the method described in Example 1.
  • Low endotoxin sodium alginate is as described in Example 1.
  • Phosphate buffer solution is sodium dihydrogen phosphate (Wako Pure Chemical Industries, Ltd., 197-09705 (trade name)), potassium dihydrogen phosphate (Wako Pure Chemical Industries, Ltd., 166-04255 (trade name)), sodium chloride (Japanese The adjustment was made using Kojun Pharmaceutical Co., Ltd., 191-01665 (trade name), and potassium chloride (Wako Pure Chemical Industries, Ltd., 166-17945 (trade name)).
  • Sodium ethylenediaminetetraacetate (N001) was purchased from Dojindo.
  • the fluorescently modified laminated sponge was punched out with a biopsy tray (diameter: 8 mm, Kai Medical BP-60F (trade name)). This was immersed in 10 mL of 150 mM phosphate buffer (pH 7.5), and 200 ⁇ L of the immersion liquid was collected at regular intervals. The recovered solution was transferred to a 96-well plate, and the amount of dissolved alginic acid was quantified by measuring the fluorescence intensity with a fluorescence plate reader.
  • the ratio of the elution amount of the monovalent metal salt of alginic acid in the lower layer is 36% (less than 50%) at 1 hour from the start of measurement 2.) 55% (less than 70%) at 2 hours.
  • the dissolution rate of the upper layer was faster than that of the lower layer.
  • Example 3 Rat Partial Hepatectomy Model The formation of adhesions was evaluated using a rat partial hepatectomy model.
  • the rat partial hepatectomy model is a model that can cause severe inflammation and observe formation of adhesions with high strength with high reproducibility (Shimizu A et al., (2014) Surg Today. (44): 314- 323). Specifically, formation of adhesions was evaluated as follows.
  • Seprafilm (trade name) is a sheet-like material in which carboxymethylcellulose (CMC) and hyaluronic acid are mixed, and was obtained from Genzyme GmbH.
  • Interceed (trade name) is a regenerated oxidized cellulose sheet, obtained from Johnson & Johnson.
  • 500-10 laminated sponge group (n 8): The alginate sponge prepared in Example 1 was applied as an adhesion prevention material.
  • Seprafilm group (n 8): 2 ⁇ 3 cm of Seprafilm was applied as anti-adhesion material.
  • Interceed group (n 8): 2 ⁇ 3 cm Interceed was applied as an anti-adhesion material.
  • the rats were anesthetized by intraperitoneal administration of pentobarbital equivalent to 35 mg / kg, and the body weights were measured by an electronic balance. The rats were then opened at midline incision. Next, the abdominal wall was pinched and lifted with tweezers, and the abdominal wall was cut. In preparation for performing hepatectomy, the left lateral lobe was pulled out of the back of the abdominal cavity and a gauze was placed under it. Next, I moved to actual liver resection. Specifically, a scale was applied to the liver to find a position where the separation plane was 3 cm, and bipolar cauterization and marking were made on both ends. A linear cut was made between the two points marked. In the Control group, treatment was terminated immediately after this.
  • the anti-adhesion agent was then applied after removing the gauze.
  • the abdominal wall and the skin were sutured in two portions and closed.
  • biodegradable yarn was used, and when sutured the skin, non-absorbable yarn was used.
  • the rats were euthanized with an overdose of approximately 2 mL of pentobarbital as an excess of anesthesia and the weight was measured by electronic balance. Thereafter, the abdomen was reopened and adhesions were evaluated as follows. The spleen weight was measured with an electronic balance after the spleen was removed from the abdominal cavity.
  • Adhesions were evaluated as follows.
  • Non-separating section The following (a) and (b) were evaluated for liver surface, greater omentum, peritoneum, small intestine, just below midline wound, etc. other than liver separation section.
  • (A) Adhesion grade The adhesion was evaluated by visual observation. The adhesion score was assigned to the site other than the liver dissection according to the following scoring method. The site was not identified, and the maximum adhesion score observed was recorded as the adhesion score of the test animal. Adhesion score: Grade 0: no adhesions are seen at all.
  • Grade 1 adhesion that detaches by its own weight (physiological adhesion)
  • Grade 2 adhesion that can be exfoliated by tweezers (blunt adhesion)
  • Grade 3 scissors, adhesions that can not be exfoliated without using a scalpel (sharp adhesions)
  • Example 3-2 The same material, experimental group, procedure and adhesion evaluation method as in Example 3 were used except that pean forceps were used in transection of a rat partial hepatectomy model liver using pean forceps. The adhesion grade and adhesion Extent of liver dissection and non-cross-section were evaluated using.
  • the transection of the liver using Peean forceps was performed as follows. That is, in [Procedure of Example 3], “cut linearly between the two marked points” was performed by crushing the liver parenchyma with a pean forceps and cauterizing the exposed blood vessel with a bipolar .
  • the AL10 layer is widely distributed also on the surface of the peritoneum in addition to the dissection. From this, it is suggested that the second layer of the sponge-like laminate rapidly dissolves and spreads in the abdominal cavity because the weight average molecular weight of the monovalent metal salt of alginic acid is relatively low.
  • the AL 500 layer was partially seen in the abdominal wall etc., the fluorescence from the cross section was more prominently observed. This suggests that the first layer of the sponge-like laminate has a relatively high weight-average molecular weight of the monovalent metal salt of alginic acid, and therefore, remains in the cross section to form a physical barrier.
  • Example 5 Winding Test In order to show the adhesion followability to the curved surface of the alginate laminate sponge, a winding test was conducted as follows.
  • Low endotoxin sodium alginate is as described in Example 1.
  • Agar (010-08725) was purchased from Wako Pure Chemical Industries.
  • the anti-adhesion material including the sponge-like laminate can be used for intestinal anastomosis and the like.
  • Example 6 Sponge Pressing and Swelling Test The alginate-laminated sponge was pressed, the thickness after pressing was measured, and the swelling test was conducted as follows.
  • the alginate-laminated sponge (AL10 (upper layer)-AL 500 (lower layer)) is as described in Example 1.
  • the average thickness of the sponge was about 1.5 mm before pressing, about 0.33 mm after manual pressing, and about 0.16 mm after pressing by a pressing machine. The thickness of the pressed sponge was maintained without increasing in thickness over time.
  • Example 7 Fog Spray Test The following test was conducted to evaluate the brittleness at the time of water absorption for the alginate laminate sponge and Seprafilm (trade name).
  • the alginate-laminated sponge is as described in Example 1 and the Seprafilm (trade name) as described in Example 3. Moreover, what was pressed with the press of Example 6 was used as a thing with a press of an alginate lamination
  • Test pieces of 1 cm ⁇ 2 cm were prepared from the alginate-laminated sponge and Seprafilm (trade name).
  • a double-sided tape was attached to one end 1 cm ⁇ 1 cm of the test piece, and was held at the end of the test stand. Thereby, the test piece was fixed so that the other end 1 cm ⁇ 1 cm was hollow. Pure water was sprayed five times onto each test piece using a mist spray.
  • a moving image was taken of the process in which the test piece bent downward as the sample wetted. By image analysis of the obtained moving image, both the height and angle from the test stand of the test piece tip were calculated, and the time change was plotted.
  • Example 8 Preparation of Pharmaceutical Component-Containing Alginate Sponge
  • a pharmaceutical component-containing alginate sponge was prepared as follows. [reagent] The respective reagents used for the preparation of the pharmaceutical component-containing alginate sponge are as follows. Low endotoxin sodium alginate was obtained from Mochida Pharmaceutical Co., Ltd. AL500: (Lot NO. 5F05204), endotoxin amount 19 EU / g. Calcium chloride, dexamethasone (Dex) and dexamethasone sodium phosphate ester (DSP) were obtained from Wako Pure Chemical Industries, Ltd. -Calcium chloride: (Product code: 036-00485). Dex: (Product Code: 040-30811). DSP: (Product Code: 041-18861 ").
  • the obtained alginic acid sponge was substantially circular, having the same diameter and thickness as the laminated sponge obtained in Example 1.
  • 500 ⁇ L of the Dex solution was dropped to the prepared alginic acid sponge, and ethanol was volatilized by standing to obtain an alginic acid sponge containing 5 mg of Dex.
  • A-2) Alginic Acid Sponge Containing DSP In the same manner as in A-1), an alginic acid sponge was prepared using AL500. 200 ⁇ L of the DSP solution was added dropwise to the prepared alginic acid sponge, and water was removed by freezing again at ⁇ 20 ° C. for 4 hours and drying to obtain alginic acid sponge containing 5 mg of DSP.
  • Dex-containing alginic acid sponge 1) 1 wt% alginic acid solution is prepared by the method of Example 1 [Preparation procedure] (1) did. 3 mL of the Dex solution prepared in (1) i) above was added to 6 mL of the 1 wt% alginic acid solution, and mixed well. 2) 600 ⁇ L of a 100 mM calcium chloride solution was added to 2.4 mL of pure water and mixed well. 3) The two solutions were well mixed in a 10 mL syringe to obtain a Dex-containing alginate gel.
  • the alginic acid gel obtained in 3) was dispensed into petri dishes in 2 mL portions, frozen at ⁇ 20 ° C., and dried in a lyophilizer for 4 hours to obtain an alginic acid sponge containing 5 mg of Dex.
  • the obtained alginic acid sponge was substantially circular, having the same diameter and thickness as the laminated sponge obtained in Example 1.
  • another Dex-containing alginate sponge was produced in the same manner as described above except that the two solutions were mixed using a pipette on a petri dish without using a syringe.
  • a 1 wt% alginic acid solution was prepared by the method of Example 1 [Preparation procedure] (1). 1.2 mL of the DSP solution prepared in (1) ii) above was added to 6 mL of a 1 wt% alginic acid solution and mixed well. 2) 600 ⁇ L of a 100 mM calcium chloride solution was added to 4.2 mL of pure water and mixed well. 3) The two solutions were well mixed in a 10 mL syringe to obtain a DSP-containing alginate gel.
  • the alginic acid gel obtained in 3) was dispensed into petri dishes in 2 mL portions, frozen at ⁇ 20 ° C., and dried in a lyophilizer for 4 hours to obtain an alginic acid sponge containing 5 mg of DSP.
  • the obtained alginic acid sponge was substantially circular, having the same diameter and thickness as the laminated sponge obtained in Example 1.
  • another DSP-containing alginate sponge was produced in the same manner as described above except that the two solutions were mixed using a pipette on a Petri dish without using a syringe.
  • Alginate sponges prepared by instillation of Dex or DSP showed no difference in appearance from alginate sponges containing no pharmaceutical ingredient.
  • alginic acid sponges prepared by mixing Dex or DSP with alginic acid gel gave different results when mixing with a pipette on a petri dish and when mixing well in a syringe. Since what was mixed on the petri dish started gelation immediately after pouring 2 liquids on the petri dish and it was difficult to mix, mixing became inadequate. As a result, a sponge having an uneven crosslinking density was obtained. On the other hand, when well mixed and gelled using two syringes, a uniform gel was obtained, and as a result, a sponge having a uniform crosslink density was obtained.
  • Example 9 Rat 70% Hepatectomy Model In order to evaluate the adhesion-preventing effect in vivo, the rat 70% liver resection model (Shimizu et al. Surgery Today (2014) 44, pp. 314-323) was used. . From the results of using the rat partial hepatectomy model described in Examples 3 and 3-2, the adhesion preventing effect of the alginate laminate sponge is confirmed. In this example, the adhesion preventing effect was evaluated using a 70% liver resection model in which the degree of invasiveness is higher than the rat partial liver resection models in Examples 3 and 3-2 and more severe adhesion is caused.
  • a monolayer sponge and a pharmaceutical component-containing alginate-laminated sponge were prepared as follows.
  • AL500 single layer sponge was prepared by the method described in Example 8. 1) Preparation of a Layered Sponge Containing Dex in the First Layer (Lower Layer) and Having No Pharmaceutical Ingredient in the Second Layer (Upper Layer): The Dex-containing alginate gel obtained by the method of Example 8, B-1), 1) to 3) was fractionated in a 2 mL petri dish and frozen at ⁇ 20 ° C. for 4 hours.
  • Example 1 [Preparation procedure] (3) and (4), a laminated sponge containing Dex in the first layer was obtained.
  • Example 1 Preparation Procedure
  • an AL500 layer (lower layer) is formed according to (1) and (2), a 1% solution of AL10 is prepared according to the method of Example 8, B-1), 1) to 3).
  • the Dex-containing alginate gel prepared using was laminated to the AL500 layer and frozen at -20.degree. C. for 4 hours.
  • Example 1 [Preparation procedure] (4) a laminated sponge containing Dex in the second layer was obtained.
  • Example 1 Preparation of a Layered Sponge Containing DSP in the Second Layer (Top Layer) without Containing a Pharmaceutical Component in the First Layer (Low Layer):
  • Example 1 [Preparation Procedure] After an AL500 layer (lower layer) is formed according to (1) and (2), a 1% solution of AL10 according to the method of Example 8, B-2), 1) to 3) The DSP-containing alginate gel prepared using was laminated to the AL500 layer and frozen at -20.degree. C. for 4 hours.
  • Single layer sponge group ⁇ AL 500 group (n 8): After performing the same treatment as the Control group, attach a sponge made of 2 cm ⁇ 2 cm 1 wt% alginic acid (AL 500) solution and 10 mM calcium chloride solution I was closed.
  • Interceed group (n 8): After performing the same treatment as the Control group, 2 cm ⁇ 2 cm Interceed was attached to the liver separation section and the abdomen was closed.
  • Layered sponge group Lower layer: Dex group (n 8): After performing the same treatment as the Control group, a 2 cm ⁇ 2 cm Dex-containing layered sponge was attached to the liver separation section and the abdomen was closed.
  • the laminated sponge lower layer AL500 contains Dex.
  • Lower layer: DSP group (n 8): After performing the same treatment as the Control group, a 2 cm ⁇ 2 cm DSP-containing laminated sponge was attached to the liver separation section and the abdomen was closed.
  • the laminated sponge lower layer AL500 contains a DSP.
  • Upper layer: Dex group (n 8): After performing the same treatment as the Control group, a 2 cm ⁇ 2 cm Dex-containing laminated sponge was attached to the liver separation section and the abdomen was closed.
  • the laminated sponge upper layer AL10 contains Dex.
  • Upper layer: DSP group (n 8): After performing the same treatment as in the Control group, a 2 cm ⁇ 2 cm Dex-containing laminate sponge was attached to the liver separation section and the abdomen was closed.
  • the laminated sponge upper layer AL10 contains a DSP.
  • Rats were anesthetized by somnopentyl intraperitoneal administration. Thereafter, the abdominal wall of the rat was cut at midline incision. In preparation for liver resection, the non-absorbable thread was lightly tied into a loop, and the rat liver was lifted and fixed to pass through the looped thread. The thread was moved gradually to the root of the liver while being tied gradually, and moved to the root of the liver. The thread was tied tightly to tie down the underlying blood vessels of the liver. The liver was then excised near the knot of the thread. The other liver was resected following the same procedure. 10 ml of physiological saline was poured from the syringe into the abdominal cavity and washed.
  • FIGS. 11 (A) and 11 (B) show the evaluation results of the presence or absence of adhesion and the length of adhesion on the liver separation cross section-diaphragm (wound surface (cross section)). Moreover, the evaluation result of adhesion presence or absence and adhesion length in liver-stomach (non-cross section) is shown in FIG. 12 (A) and (B).
  • the adhesion preventing effect in the separation plane was recognized by the laminated sponge containing the pharmaceutical component.
  • the adhesion prevention effect is high with the laminated sponge containing the pharmaceutical component in the upper layer, and in particular, a significant adhesion prevention effect was observed in the laminated sponge containing the DSP in the upper layer.
  • an abscess was observed around the application site in part of the group using the layered sponge containing Dex or DSP in the lower layer, which was assumed to be an adverse event caused by dexamethasone. No abscess was observed in the layered sponge containing Dex or DSP in the upper layer.
  • the adhesion preventing effect was recognized even in the non-separating section by the laminated sponge containing the pharmaceutical component. It was confirmed that a higher adhesion prevention effect can be obtained when using the laminated sponge containing Dex or DSP in the upper layer, as compared to the laminated sponge containing Dex or DSP in the lower layer.
  • Example 10 Measurement of Dissolution Rate of Pharmaceutical Ingredient from Sponge in Phosphate Buffer (PBS) Dexamethasone (AL 10 monolayer sponge and AL 500 monolayer sponge) according to the method of Example 8, B) The sponge (4 types in total) which contained Dex) and dexamethasone sodium phosphate (DSP) was manufactured, and the measurement of the elution rate of the pharmaceutical component from each sponge was performed.
  • PBS Phosphate Buffer
  • DSP dexamethasone sodium phosphate
  • the low endotoxin sodium alginate is as described in Example 1, the phosphate buffer as in Example 2, and the dexamethasone and dexamethasone sodium phosphate ester as in Example 9.
  • a Dex-containing AL10 sponge, a DSP-containing AL10 sponge, a Dex-containing AL500 sponge, and a DSP-containing AL500 sponge were respectively prepared according to the method of Example 8, B).
  • Each sponge was punched out with a biopsy test pan (Kai medical BP-60F (trade name)) with a diameter of 8 mm. This was immersed in 15 mL of 150 mM phosphate buffer (pH 7.5, 37 ° C.), and 500 ⁇ L each of the immersion liquid was collected before, 30 minutes, 1 hour, 2 hours, 4 hours and 6 hours after immersion .
  • the absorbance at ⁇ 242 nm of the collected solution was measured to evaluate the dissolution rate of the drug component.
  • the pharmaceutical component contained in the sponge corresponding to the second layer of the laminated sponge is rapidly eluted regardless of the lipid solubility / water solubility. Therefore, it is assumed that it is preferable that the second layer contains a pharmaceutical component which is expected to have a pharmacological effect in the early postoperative period (for example, in the early stage of post-operative inflammation). On the other hand, it is assumed that it is preferable for the first layer to be preferably contained after confirming the in vitro dissolution behavior for a pharmaceutical component that is expected to have a sustained pharmacological effect locally for a predetermined period after surgery.

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  • Engineering & Computer Science (AREA)
  • Materials Engineering (AREA)
  • Composite Materials (AREA)
  • Biomedical Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Materials For Medical Uses (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicinal Preparation (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

L'invention concerne un matériau empêchant les adhérences qui comprend un stratifié spongiforme stérilisé qui peut être appliqué à des corps vivants et qui contient une première couche spongiforme et une seconde couche d'un sel métallique monovalent de l'acide alginique au moins partiellement réticulé avec un durcisseur. Le poids moléculaire moyen en poids du sel métallique monovalent de l'acide alginique dans la première couche se situe dans la plage allant de 10 000 à 2 000 000, et le poids moléculaire moyen en poids du sel métallique monovalent de l'acide alginique dans la seconde couche se situe dans la plage allant de 1 000 à 1 000 000. Le poids moléculaire moyen en poids est mesuré par GPC-MALS après dé-réticulation. Le poids moléculaire moyen en poids du sel métallique monovalent de l'acide alginique dans la première couche est supérieur au poids moléculaire moyen en poids du sel monovalent de l'acide alginique dans la seconde couche. La première couche et/ou la seconde couche contiennent au moins un ingrédient pharmaceutique choisi dans le groupe constitué par un médicament, un facteur de croissance, une hormone, une protéine ou une combinaison de ceux-ci. En adoptant cette configuration, il est possible de fournir un matériau empêchant les adhérences, avec un fort effet empêchant l'adhésion.
PCT/JP2018/041770 2017-11-13 2018-11-12 Composition pour empêcher les adhérences WO2019093505A1 (fr)

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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH11253547A (ja) * 1998-03-11 1999-09-21 Kunio Ishikawa 細胞遮断膜
JP2007538125A (ja) * 2004-05-21 2007-12-27 ドクター．スェラック スキン アンド ヘルス ケアー アーゲー アルギネートを含む多孔性成型品の製造方法
US20120039959A1 (en) * 2010-08-16 2012-02-16 Joerg Tessmar Anti-Adhesion Alginate Barrier of Variable Absorbance
WO2016114355A1 (fr) * 2015-01-15 2016-07-21 国立大学法人東京大学 Composition pour prévenir les adhésions
WO2018012605A1 (fr) * 2016-07-13 2018-01-18 持田製薬株式会社 Composé empêchant l'adhésion.

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH11253547A (ja) * 1998-03-11 1999-09-21 Kunio Ishikawa 細胞遮断膜
JP2007538125A (ja) * 2004-05-21 2007-12-27 ドクター．スェラック スキン アンド ヘルス ケアー アーゲー アルギネートを含む多孔性成型品の製造方法
US20120039959A1 (en) * 2010-08-16 2012-02-16 Joerg Tessmar Anti-Adhesion Alginate Barrier of Variable Absorbance
WO2016114355A1 (fr) * 2015-01-15 2016-07-21 国立大学法人東京大学 Composition pour prévenir les adhésions
WO2018012605A1 (fr) * 2016-07-13 2018-01-18 持田製薬株式会社 Composé empêchant l'adhésion.

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