Preventive or therapeutic agent for pancreatic fistula

Abstract

The present invention provides: an agent which is for preventing or treating pancreatic leak and / or pancreatic fistula after abdominal surgery and contains a lipase inhibitor as an active ingredient; and a method which is for preventing or treating pancreatic leak and / or pancreatic fistula after abdominal surgery and comprises administering an effective amount of a lipase inhibitor to a mammal in need of the administration thereof.

Description

[Technical Field]

[0001] The present invention relates to an agent for preventing or treating pancreatic leakage or fistula after abdominal surgery, and a method for preventing or treating pancreatic leakage or fistula after abdominal surgery, and is useful in the field of medicine. [Background technology]

[0002] Pancreatic surgery is widely performed clinically as the only curative treatment for various pancreatic tumor lesions. After abdominal surgery, including pancreatic surgery, pancreatic juice may leak from the pancreas due to surgical injury. Pancreatic fistula is a serious complication following abdominal surgery caused by damage to surrounding tissues by leaked pancreatic juice after surgery, and it poses a major challenge to the safe performance of pancreatic surgery and abdominal surgery around the pancreas.

[0003] Pancreatic fistulas are classified into biochemical leaks, which do not have clinically harmful effects, and clinically relevant postoperative pancreatic fistulas (CR-POPFs), which are severe and require various treatments (Non-Patent Document 1). However, the reason why postoperative pancreatic leaks remain biochemical leaks in some patients while others progress to severe CR-POPF remains unknown.

[0004] Therefore, a specific drug treatment for pancreatic leakage or fistula after abdominal surgery has not yet been established, and its establishment has been eagerly awaited. [Prior art documents] [Non-patent literature]

[0005] [Non-Patent Document 1] Surgery, 161(2017)584-591 Summary of the Invention [Problem to be solved by the invention]

[0006] As described above, it cannot be said that methods for preventing or treating pancreatic leakage and fistula after abdominal surgery have yet been established, and the development of clinically effective preventive or therapeutic agents has been an urgent issue in the field of medicine. [Means for solving the problem]

[0007] Under these circumstances, the present inventors have conducted extensive research and have found for the first time that lipase inhibitors are useful for preventing or treating pancreatic leakage and / or pancreatic fistula after abdominal surgery, leading to the completion of the present invention. Furthermore, they have also discovered a formulation containing a lipase inhibitor that is suitable for administration to patients and overcomes the problem of poor water solubility of lipase inhibitors. That is, the present invention is as follows.

[0008] [1] A preventive or therapeutic agent for pancreatic leakage and / or pancreatic fistula after abdominal surgery, which contains a lipase inhibitor as an active ingredient. [2] The preventive or therapeutic agent according to [1] above for preventing or treating pancreatic fistula after abdominal surgery. [3] The preventive or therapeutic agent according to the above [2], wherein the pancreatic fistula is a clinically relevant postoperative pancreatic fistula (CR-POPF). [4] The preventive or therapeutic agent according to any one of the above [1] to [3], wherein the abdominal surgery is pancreatic surgery.

[0009] [5] The preventive or therapeutic agent according to any one of the above [1] to [4], wherein the lipase inhibitor is one or a combination of two or more drugs selected from orlistat, cetilistat, raristat, and atglistatin. [6] The preventive or therapeutic agent according to the above-mentioned [5], wherein the lipase inhibitor is a combination of one or two drugs selected from orlistat and cetilistat.

[0010] [7] The preventive or therapeutic agent according to any one of the above [1] to [6], which contains a lipase inhibitor in an amount of 0.01 to 20% (w / w).

[0011] [8] The preventive or therapeutic agent according to any one of the above [1] to [7], which is for intraperitoneal administration (preferably, intraperitoneal local administration).

[0012] [9] The preventive or therapeutic agent according to any one of the above [1] to [8], which is an emulsified preparation containing a lipase inhibitor and a dispersant.

[0013]

[10] The preventive or therapeutic agent according to any one of the above [1] to [8], which is a hydrogel preparation containing a complex of a lipase inhibitor and a hydrogel.

[11] The preventive or therapeutic agent according to

[10] above, wherein the hydrogel is formed from any one of the following combinations of hydrogel-forming components: 1) A combination of a hydrophilic biocompatible polymer and a biocompatible hydrophilic polymer crosslinker; 2) A combination of a synthetic biocompatible polymer with functional groups (polymer A) and a synthetic biocompatible polymer with functional groups that react with polymer A under in vivo conditions (polymer B); or 3) Combination of functionalized small molecule compounds and biocompatible hydrophilic polymeric crosslinkers.

[12] The preventive or therapeutic agent according to the above

[11] , wherein the synthetic biocompatible polymer contains a polyalkylene oxide having a functional group.

[13] The preventive or therapeutic agent according to

[11] above, wherein the hydrophilic biocompatible polymer is at least one selected from gelatin, collagen, fibrinogen, fibrin, cellulose, chitosan, and derivatives thereof (more preferably at least one selected from cross-linked gelatin, fibrinogen, fibrin, and derivatives thereof, particularly preferably at least one selected from fibrinogen, fibrin, and derivatives thereof).

[14] The preventive or therapeutic agent according to any one of the above

[10] to

[13] , which contains 0.01 to 10% (W / W) of a lipase inhibitor.

[15] The preventive or therapeutic agent according to the above

[14] , which contains a lipase inhibitor in an amount of 0.01 to 5% (weight of lipase inhibitor / weight of hydrogel-forming component).

[16] The preventive or therapeutic agent according to any one of the above

[10] to

[15] , further comprising a dispersing agent.

[0014]

[17] A method for preventing or treating pancreatic leakage and / or fistula after abdominal surgery, comprising administering an effective amount of a lipase inhibitor to a mammal in need thereof.

[18] The method for preventing or treating pancreatic fistula after abdominal surgery according to the above

[17] .

[19] The method for prevention or treatment according to

[18] above, wherein the pancreatic fistula is a clinically relevant postoperative pancreatic fistula (CR-POPF).

[20] The method for prevention or treatment according to any one of the above

[17] to

[19] , wherein the abdominal surgery is pancreatic surgery.

[0015]

[21] The method for prevention or treatment according to any one of the above

[17] to

[20] , wherein the lipase inhibitor is one or a combination of two or more drugs selected from orlistat, cetilistat, ralistat and atglistatin.

[22] The method for prevention or treatment according to the above

[21] , wherein the lipase inhibitor is a combination of one or two drugs selected from orlistat and cetilistat.

[0016]

[23] The method for prevention or treatment according to any one of the above

[17] to

[22] , wherein the lipase inhibitor is administered in the form of a pharmaceutical composition containing the lipase inhibitor in an amount of 0.01 to 20% (w / w).

[0017]

[24] The method for prevention or treatment according to any one of the above-mentioned

[17] to

[23] , wherein the lipase inhibitor is administered intraperitoneally (preferably, locally administered intraperitoneally).

[0018]

[25] The method for prevention or treatment according to any one of the above

[17] to

[24] , wherein the lipase inhibitor is administered in the form of an emulsified preparation containing the lipase inhibitor and a dispersing agent.

[0019]

[26] The method for prevention or treatment according to any one of the above

[17] to

[24] , wherein the lipase inhibitor is administered in the form of a hydrogel preparation containing a complex of the lipase inhibitor and a hydrogel.

[27] The method for prevention or treatment according to

[26] above, wherein the hydrogel is formed from any one of the following combinations of hydrogel-forming components: 1) A combination of a hydrophilic biocompatible polymer and a biocompatible hydrophilic polymer crosslinker; 2) A combination of a synthetic biocompatible polymer with functional groups (polymer A) and a synthetic biocompatible polymer with functional groups that react with polymer A under in vivo conditions (polymer B); or 3) Combination of functionalized small molecule compounds and biocompatible hydrophilic polymeric crosslinkers.

[28] The method for prevention or treatment according to the above

[27] , wherein the synthetic biocompatible polymer contains a polyalkylene oxide having a functional group.

[29] The preventive or therapeutic method according to

[27] above, wherein the hydrophilic biocompatible polymer is at least one selected from gelatin, collagen, fibrinogen, fibrin, cellulose, chitosan, and derivatives thereof (more preferably at least one selected from cross-linked gelatin, fibrinogen, fibrin, and derivatives thereof, particularly preferably at least one selected from fibrinogen, fibrin, and derivatives thereof).

[30] The method for prevention or treatment according to any one of the above

[26] to

[29] , wherein the hydrogel preparation contains 0.01 to 10% (W / W) of a lipase inhibitor.

[31] The method for prevention or treatment according to the above

[30] , wherein the hydrogel formulation contains 0.01 to 5% of a lipase inhibitor (weight of lipase inhibitor / weight of hydrogel-forming component).

[32] The method for prevention or treatment according to any one of the above

[26] to

[31] , wherein the hydrogel formulation further contains a dispersing agent.

[0020]

[33] A hydrogel formulation containing a complex of a lipase inhibitor and a hydrogel.

[34] The hydrogel formulation according to

[33] above, wherein the hydrogel is formed from any one of the following combinations of hydrogel-forming components: 1) A combination of a hydrophilic biocompatible polymer and a biocompatible hydrophilic polymer crosslinker; 2) A combination of a synthetic biocompatible polymer with functional groups (polymer A) and a synthetic biocompatible polymer with functional groups that react with polymer A under in vivo conditions (polymer B); or 3) Combination of functionalized small molecule compounds and biocompatible hydrophilic polymeric crosslinkers.

[35] The hydrogel formulation according to

[34] above, wherein the synthetic biocompatible polymer contains a polyalkylene oxide having a functional group.

[36] The hydrogel formulation according to

[34] above, wherein the hydrophilic biocompatible polymer is at least one selected from gelatin, collagen, fibrinogen, fibrin, cellulose, chitosan, and derivatives thereof (more preferably at least one selected from cross-linked gelatin, fibrinogen, fibrin, and derivatives thereof, particularly preferably at least one selected from fibrinogen, fibrin, and derivatives thereof).

[37] The hydrogel preparation according to any one of the above

[33] to

[36] , which contains 0.01 to 10% (W / W) of a lipase inhibitor.

[38] The hydrogel formulation according to

[37] above, containing 0.01 to 5% of a lipase inhibitor (weight of lipase inhibitor / weight of hydrogel-forming component).

[39] The hydrogel preparation according to any one of the above

[33] to

[38] , further comprising a dispersing agent. [Effects of the Invention]

[0021] According to the present invention, it is possible to provide a drug that is effective and safe to use in the prevention or treatment of pancreatic leakage or pancreatic fistula after abdominal surgery; and a method for preventing or treating pancreatic leakage or pancreatic fistula after abdominal surgery using the drug. [Brief explanation of the drawings]

[0022] [Figure 1] Figure 1 shows a schematic diagram of the preparation of pancreatic fistula model mice (PT) and fat injury treatment (+F). [Figure 2] FIG. 2 shows the intraperitoneal findings (24 hours after treatment) for each group examined in Example 1(1). [Figure 3] Figure 3 shows the ascites free fatty acid concentration of each group studied in Example 1(2). A Tukey test was used to test for significance between the sham group and the group. In the figure, TFA stands for total free fatty acids, SFA stands for saturated free fatty acids, and UFA stands for unsaturated free fatty acids. Specific values for TFA concentration (µM) are as follows: Sham group: 263, Sham+F group: 390, PT group: 528, PT+F group: 4145. [Figure 4] Figure 4 shows the pancreatic enzyme activity (IU / L) in the ascites for each group studied in Example 1(3). The specific activity of each enzyme is as follows: Amylase: Sham group: 34, Sham+F group: 104, PT group: 2468, PT+F group: 16868*. The PT+F group had significantly higher enzyme activity than the PT group (p=0.009; Mann-Whitney U test). Lipase: Sham group: 0, Sham+F group: 2, PT group: 46, PT+F group: 2017*. The PT+F group had significantly higher enzyme activity than the PT group (p=0.009; Mann-Whitney U test). [Figure 5]Figure 5 shows the histological findings of each group examined in Example 1 (4). Compared to the Sham group, no obvious changes were observed in the pancreas in the Sham+F group. In the PT group, mild inflammatory changes were confirmed, including mild interstitial edema and cellular infiltration. In addition to these changes, the PT+F group also showed deposition of hyaline-like material on the pancreatic surface. This area stained orange with alizarin staining, indicating that it was fatty acid calcium deposition. High levels of inflammatory cell infiltration were observed around the hyaline-like deposits, and signs of acinar cell damage were observed. [Figure 6] FIG. 6 shows the survival rates in the PT group and the PT+F group examined in Example 1(5). [Figure 7] FIG. 7 shows the effect of neutral fat on rat pancreatic acinar cells examined in Example 2(1). [Figure 8] 8 shows the protective effect of PEG-C on pancreatic acinar cells examined in Example 2(2). In the figure, C20 refers to the group administered with 20 μM PEG-C, C100 refers to the group administered with 100 μM PEG-C, None refers to the group not administered with PEG-C, and PEG refers to the group administered with an aqueous PEG solution (equivalent to 100 μM PEG-C). [Figure 9] FIG. 9 shows the intraperitoneal findings (24 hours after treatment) for each group examined in Example 3(1). [Figure 10] Figure 10 shows the pancreatic enzyme activity (IU / L) in the ascites for each group studied in Example 3(2). The specific activity of each enzyme is as follows: Amylase: PT group: 2468, PT+F group: 16868, PT+F+C group: 4374, PT+F+V group: 30120*. The PT+F group had significantly higher enzyme activity than the PT+F+C group (p=0.009; Mann Whitney U test). Lipase: PT group: 46, PT+F group: 2017; PT+F+C group: 573, PT+F+V group: 7725*. The PT+F group had significantly higher enzyme activity than the PT+F+C group (p=0.079; Mann Whitney U test). [Figure 11]Figure 11 shows the ascites free fatty acid concentration (μM) in each group studied in Example 3(3). In the figure, TFA means total free fatty acids, SFA means saturated free fatty acids, and UFA means unsaturated free fatty acids. Specific values for TFA concentration (μM) are as follows: PT group: 471, PT+F group: 4172, PT+F+C group: 1294, PT+F+V group: 2877. *The PT+F group had significantly higher enzyme activity than the PT+F+C group (p=0.016; Mann Whitney U test). [Figure 12] FIG. 12 shows the survival rate of each group examined in Example 3(4). [Figure 13] Figure 13 shows the histological findings in the PEGgel-C administration group examined in Example 5(1). Although a small amount of saponified material was observed around the pancreas (around the hydrogel), overall saponification was inhibited. Compared to the untreated group (PT+F group), saponification was clearly inhibited. [Figure 14] Figure 14 shows the histological findings in the PEGgel-C administration group examined in Example 5(1). Although a small amount of saponified material was observed around the pancreas (around the hydrogel), overall saponification was inhibited. Compared to the untreated group (PT+F group), saponification was clearly inhibited. [Figure 15] Figure 15 shows the histological findings in the PEGgel-administered group examined in Example 5(1). Extensive saponification and hemolysis were observed, suggesting a worsening of the condition. [Figure 16] 16 shows histological findings in the PEGgel-administered group examined in Example 5(1). Noticeable saponification was confirmed. [Figure 17] 17 shows the histological findings of the untreated group (PT+F group) compared with the histological findings of the PEGgel-C group / PEGgel group in Example 5(1). Extensive saponification was observed in the abdominal cavity. [Figure 18]Figure 18 shows the results of the investigation of pancreatic enzyme activity (IU / L) in ascites in Example 5(2). In the figure, ascites AMY indicates the enzyme activity of amylase, and ascites Lipase indicates the enzyme activity of lipase. Amylase: PT+F group: 16868, PEGgel-C group: 1278 (p=0.025; Mann-Whitney U test). Lipase: PT+F group: 2017, PEGgel-C group: 52 (p=0.025; Mann-Whitney U test). [Figure 19] Figure 19 shows the histological findings in the PEGgel-O administration group examined in Example 7(1). In the PEGgel-O group, a clear reduction (inhibition) was confirmed in both the overall inflammatory findings and saponification in the abdominal cavity compared to the untreated group (see Figure 17). [Figure 20] Figure 20 shows the histological findings in the Bolheal-C administration group examined in Example 9(1). In the Bolheal-C group, a clear reduction (inhibition) was confirmed in both the overall inflammatory findings in the abdominal cavity and the evaluation items of saponification, compared to the untreated group (see Figure 17). [Figure 21] Figure 21 shows the histological findings in the group administered with Bolheal alone (Comparative Example 2(1)). In the group administered with Bolheal alone, inflammation and saponification were observed throughout the abdominal cavity, similar to the untreated group (see Figure 17). DETAILED DESCRIPTION OF THE INVENTION

[0023] The present invention relates to an agent for preventing or treating pancreatic leakage and / or fistula after abdominal surgery, which contains a lipase inhibitor as an active ingredient; a method for preventing or treating pancreatic leakage and / or fistula after abdominal surgery, which comprises administering an effective amount of a lipase inhibitor to a mammal in need thereof; a lipase inhibitor for use in preventing or treating pancreatic leakage and / or fistula after abdominal surgery; use of a lipase inhibitor for producing an agent for preventing or treating pancreatic leakage and / or fistula after abdominal surgery; and pharmaceutical compositions containing a lipase inhibitor that are useful for carrying out the present invention.

[0024] The present invention will now be described in detail, and unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this invention belongs.

[0025] (About lipase inhibitors) As used herein, "lipase inhibitor" is a general term for compounds that have the effect of inhibiting the decomposition of neutral fats into free fatty acids by lipolytic enzymes (lipases) contained in pancreatic juice. Many "lipase inhibitors" are already known in the art, and those skilled in the art recognize and use the term "lipase inhibitor" as a comprehensive term referring to this group of compounds. Suitable examples include orlistat, cetilistat, lalistat, and atglistatin, with orlistat or cetilistat being more preferred.

[0026] For example, orlistat can be produced by the method described in Japanese Patent Laid-Open No. 60-13777. Cetilistat, ralistat, and atglistatin can also be obtained by appropriate synthesis using methods known in the art by those skilled in the art.

[0027] As will be demonstrated in the Examples section below using orlistat and cetilistat as representative "lipase inhibitors," "lipase inhibitors" exert a therapeutic effect on pancreatic fistula by suppressing the breakdown of neutral fats into free fatty acids in the abdominal cavity by lipase (a lipolytic enzyme) leaked from the pancreas, based on their function of inhibiting the breakdown of neutral fats into free fatty acids.

[0028] In carrying out the present invention, the lipase inhibitor can be used in either the free form or the form of a pharmaceutically acceptable salt thereof. Those skilled in the art can carry out the present invention by appropriately selecting either form based on the properties of the individual lipase inhibitor used. Pharmaceutically acceptable salts include salts with inorganic acids such as hydrochloride, hydrobromide, sulfate, phosphate, etc.; salts with organic acids such as acetate, fumarate, oxalate, citrate, methanesulfonate, benzenesulfonate, tosylate, maleate, etc.; salts with bases such as alkali metal salts such as sodium salt, potassium salt, etc., alkaline earth metal salts such as calcium salt, etc.; salts with amino acids such as glycine salt, lysine salt, arginine salt, ornithine salt, glutamate, aspartate, etc.

[0029] (About the target diseases) In this specification, "abdominal surgery" in "pancreatic leakage or fistula after abdominal surgery" includes surgery on the pancreas, stomach, duodenum, spleen, kidney, adrenal gland, or large intestine, and in particular, pancreatic leakage and / or pancreatic fistula after pancreatic surgery or gastric surgery (particularly after pancreatic surgery) is the subject of application of the present invention. As used herein, the term "pancreatic juice leakage" refers to leakage of pancreatic juice from the pancreas into the abdominal cavity following abdominal surgery. As used herein, the term "pancreatic fistula" refers to a complication caused by damage to surrounding tissues by leaked pancreatic juice, and includes both biochemical leaks that do not have clinically harmful effects and clinically relevant postoperative pancreatic fistulas (CR-POPFs) that cause serious symptoms and require clinical treatment.

[0030] As used herein, "prevention" includes preventing the onset of a disease (either an entire condition or one or more conditions) and delaying the onset of the disease. A "prophylactically effective amount" refers to a dose of a lipase inhibitor sufficient to achieve this purpose.

[0031] As used herein, "treatment" includes curing a disease (either the entire pathology or one or more pathologies), ameliorating the disease, and inhibiting the progression of the severity of the disease. A "therapeutically effective amount" refers to a dose of a lipase inhibitor sufficient to achieve such a purpose.

[0032] (Regarding administration route) In carrying out the present invention, the lipase inhibitor may be administered to a subject in need thereof either orally or parenterally, either alone or as a pharmaceutical composition, but intraperitoneal administration is preferred, and local administration into the peritoneal cavity is particularly preferred.

[0033] (Dosage) The dosage of the lipase inhibitor used in carrying out the present invention varies depending on the subject, route of administration, and the subject's age and symptoms, but is not particularly limited. For example, when administered intraperitoneally to an adult patient (body weight approximately 40 to 80 kg, e.g., 60 kg), the dosage can be selected within the range of 1 to 1,000 mg per day. It may be administered all at once at the time of surgery, or multiple times after surgery. It is readily apparent to those skilled in the art that the dosage of the lipase inhibitor varies depending on the individual drug used, and therefore may be administered in a smaller range, for example, within the range of 1 to 500 mg or 1 to 300 mg per day. Therefore, administration modes of the lipase inhibitor within such ranges are naturally encompassed by the above-mentioned "administration within the range of 1 to 1,000 mg." The prophylactic and therapeutic agents of the present invention can be safely administered to subjects in need thereof (mammals including humans, particularly humans).

[0034] (Regarding dosage forms) In carrying out the present invention, the lipase inhibitor is used as a "prophylactic or therapeutic agent for pancreatic leakage or fistula after abdominal surgery" (hereinafter sometimes abbreviated as "the prophylactic or therapeutic agent of the present invention") either alone or in the form of a pharmaceutical composition containing the lipase inhibitor as an active ingredient together with a pharmaceutically acceptable carrier, etc. Preferably, it is used in the form of a pharmaceutical composition.

[0035] In carrying out the present invention, the pharmaceutical composition may be in a dosage form that is used in the art, such as a tablet, an emulsion, a suspension, a controlled-release formulation, a hydrogel formulation, etc. As the "pharmaceutically acceptable carrier," various carriers commonly used in the field of formulation technology can be used. The pharmaceutical composition of the present invention can be produced by adding a lipase inhibitor to the total amount of the formulation, usually in an amount of 0.001 to 20% (w / w), preferably 0.01 to 10% (w / w), although this varies depending on the dosage form, administration method, carrier, etc. The pharmaceutical composition can be produced by a method conventionally used in the field of formulation technology, depending on the form.

[0036] Preferred forms of pharmaceutical compositions for carrying out the present invention will now be described.

[0037] (1) Emulsified preparations In carrying out the present invention, one preferred embodiment of the pharmaceutical composition is an "emulsion preparation containing a lipase inhibitor and a dispersing agent." Here, examples of dispersants that can be used include polyalkylene glycols (e.g., polyethylene glycol, polyoxyethylene polyoxypropylene glycol, etc.), and ether-type nonionic surfactants (e.g., polyoxyethylene alkyl ether, polyoxyethylene alkyl phenyl ether, etc.). For example, when polyethylene glycol is used, it is preferable to use one with a molecular weight in the range of 1,000 to 20,000. In carrying out the present invention, it is preferable to use an emulsified preparation containing 0.01 to 10% of a lipase inhibitor (weight of lipase inhibitor / weight of dispersant). The "emulsion preparation containing a lipase inhibitor and a dispersant" of the present invention can be produced using a lipase inhibitor, a dispersant, and other necessary additives by a method commonly used in the art.

[0038] (2) Hydrogel formulation Another preferred embodiment of the pharmaceutical composition for carrying out the present invention is a "hydrogel preparation containing a complex of a lipase inhibitor and a hydrogel." By forming a complex of a lipase inhibitor with a hydrogel (hydrogel formulation), it is possible to prevent leakage of pancreatic juice and to allow the lipase inhibitor to act continuously in a local area within the abdominal cavity for a certain period of time (sustained-release formulation).

[0039] The hydrogel formulation of the present invention can be produced appropriately using a lipase inhibitor and a hydrogel-forming component by techniques known in the art, taking into consideration the desired duration of action, etc., and preferred hydrogel-forming components that form a hydrogel include, for example, the following combinations: 1) Combination of a hydrophilic biocompatible polymer and a biocompatible hydrophilic polymer crosslinker (hydrogel-forming component 1) 2) A combination of a synthetic biocompatible polymer with functional groups (polymer A) and a synthetic biocompatible polymer with functional groups that react with polymer A under in vivo conditions (polymer B) (hydrogel-forming component 2), or 3) A combination of a functionalized low molecular weight compound and a biocompatible hydrophilic polymer crosslinker (hydrogel-forming component 3).

[0040] Hydrogel-forming component 1 "Hydrophilic biocompatible polymers" include proteins, polysaccharides, non-biological polymers, derivatives or combinations thereof. The protein may be selected from the group consisting of gelatin, collagen, albumin, hemoglobin, fibrinogen, fibrin, casein, fibronectin, elastin, keratin, and laminin, and derivatives and combinations thereof. The polysaccharide may be selected from the group consisting of glycosaminoglycans, starches, celluloses, dextran, hemicelluloses, xylans, agaroses, alginates, and chitosans, and derivatives and combinations thereof. The non-biological polymer can be selected from the group consisting of polyacrylate, polymethacrylate, polyacrylamide, polymethacrylamide, polyethyleneimine, polyvinyl resin, polylactide-glycolide, polycaprolactone, and polyoxyethylene, and derivatives and combinations thereof. Here, the "hydrophilic biocompatible polymer" may be crosslinked. Crosslinking can be carried out by methods known in the art. The hydrophilic biocompatible polymer is not limited to, but preferably includes at least one selected from gelatin, collagen, fibrinogen, fibrin, cellulose, chitosan, and derivatives thereof, more preferably includes at least one selected from cross-linked gelatin, fibrinogen, fibrin, and derivatives thereof, and particularly preferably includes at least one selected from fibrinogen, fibrin, and derivatives thereof. Another particularly preferred example is cross-linked gelatin.

[0041] The "biocompatible hydrophilic polymer crosslinking agent" includes a hydrophilic polymer crosslinking agent, and a suitable example thereof is a polyalkylene oxide polymer containing a functional group selected from the group consisting of succinimidyl ester (-CON(COCH2)2), an NHS-ester group, an imidoester group, an aldehyde group, a carboxy group in the presence of carbodiimide, an isocyanate, THPP (beta-[tris(hydroxymethyl)phosphino]propionic acid), maleimide, a haloacetyl, a thiol group, an amino group, an azide group, an alkyne group such as an ethynyl group, a cyclic alkyne group including a cyclooctyne group, and a mixture thereof. As the hydrophilic polymer crosslinking agent having such a functional group, polyethylene glycol containing a functional group selected from succinimidyl ester (-CON(COCH2)2), aldehyde (-CHO), thiol group, amino group, azide group, alkyne group such as ethynyl group, cyclic alkyne group including cyclooctyne group, and isocyanate (-N=C=O) is particularly preferred.

[0042] In this embodiment, the "hydrophilic biocompatible polymer" is crosslinked by the "biocompatible hydrophilic polymer crosslinker" to form a network, thereby gelling and providing a hydrogel. Note that this gelation reaction may be carried out by appropriately adding other components commonly used in the art. The above manufacturing steps can be carried out by a person skilled in the art according to conventional methods for producing hydrogels in this technical field.

[0043] Hydrogel-forming component 2 The "synthetic biocompatible polymer having functional groups (polymer A)" includes polyalkylene oxides having two or more reactive groups, and the functional groups include the same functional groups as those exemplified for the hydrophilic polymer crosslinker described above. Here, the polyalkylene oxide is preferably, for example, polyethylene oxide or a derivative thereof. The "synthetic biocompatible polymer (polymer B) having a functional group that reacts with polymer A under in vivo conditions" includes a polyalkylene oxide having two or more reactive groups, and the functional group may be the same as the functional group exemplified for the hydrophilic polymer crosslinker described above. Here, the polyalkylene oxide is preferably, for example, polyethylene oxide or a derivative thereof. When using polyethylene oxide or a derivative thereof, it is preferable to use one having a molecular weight in the range of 1,000 to 80,000.

[0044] In this embodiment, a "synthetic biocompatible polymer having functional groups (polymer A)" and a "synthetic biocompatible polymer having functional groups that react with polymer A under in vivo conditions (polymer B)" are gelled by bonding to each other via their functional groups to form a network, and are provided as a hydrogel. Note that this gelling reaction may be carried out by adding other components commonly used in the art, as appropriate. The above manufacturing steps can be carried out by a person skilled in the art according to conventional methods for producing hydrogels in this technical field.

[0045] Hydrogel-forming component 3 The "low molecular weight compound having a functional group" is a compound containing two or more functional groups of one or more types, and the functional groups include the same functional groups as those exemplified for the hydrophilic polymer crosslinking agent described above. For example, amino acids or oligopeptides having an amino group, a thiol group, etc., polyamines such as spermine, diamines such as ethylenediamine and diethylenetriamine, and triamine compounds can be mentioned. The "biocompatible hydrophilic polymer crosslinking agent" includes the above-mentioned Hydrogel-forming component 1 Examples of the "biocompatible hydrophilic polymer crosslinking agent" include the same as those exemplified in

[0046] In this embodiment, the "low molecular weight compound having a functional group" and the "biocompatible hydrophilic polymer crosslinker" are crosslinked and bonded to each other to form a network, thereby gelling and providing a hydrogel. Note that this gelation reaction may be carried out by appropriately adding other components commonly used in the art. The above manufacturing steps can be carried out by a person skilled in the art according to conventional methods for producing hydrogels in this technical field.

[0047] The hydrogel formulations of the present invention can be prepared by conventional methods for preparing hydrogels, including dissolving (emulsifying or suspending), mixing, and stirring the lipase inhibitor, the hydrogel-forming components, and other components commonly used in the art. They can be used by extrusion, spraying, or as a two-component hydrogel sheet. For example, the components forming the hydrogel can be premixed as aqueous solutions and then extruded or sprayed to form a hydrogel. Alternatively, the aqueous solutions of the two components can be extruded using a mixing tip from a syringe that can be individually filled, and then sprayed to form a hydrogel. Furthermore, a hydrogel sheet can be formed in advance from the hydrogel-forming components and applied directly to the affected area. Alternatively, the hydrogel sheet can be impregnated with a lipase inhibitor and then used. Furthermore, a hydrogel sheet can be formed in advance from the hydrogel-forming components, powdered or dried by freeze-drying or other methods, and the powder can be swelled in an aqueous solution containing a lipase inhibitor to form a hydrogel sheet, which can then be applied directly to the affected area.

[0048] Alternatively, the lipase inhibitor may be previously made into an "emulsion preparation containing a lipase inhibitor and a dispersant" as described in the above section (emulsion preparation), and then the hydrogel preparation may be produced.

[0049] If the lipase inhibitor is poorly soluble in water, it can be converted into a water-soluble form by other methods known in the art (e.g., encapsulation in polymeric micelles) and then used as a "lipase inhibitor" to produce a hydrogel.

[0050] In carrying out the present invention, it is preferable to use a hydrogel preparation containing 0.01 to 5% of a lipase inhibitor (weight of lipase inhibitor / weight of hydrogel-forming component).

[0051] (Concomitant use with other drugs)

[0052] The prophylactic and therapeutic agent of the present invention can be used in combination with other drugs for the prevention or treatment of pancreatic juice leakage or pancreatic fistula after abdominal surgery, and an excellent prophylactic or therapeutic effect can be expected by the combined use with other drugs. Further, it can also be expected to reduce the dosage of other drugs by such combined therapy and reduce the side effects they have.

Examples

[0053] Hereinafter, the present invention will be described in more detail along with examples, but these examples do not limit the scope of the present invention in any way. Also, the reagents, devices, and materials used in the present invention are commercially available or can be appropriately prepared by those skilled in the art unless otherwise specifically mentioned.

[0054] [Experimental materials and experimental methods] <Experimental materials> Cetilistat was purchased from Tokyo Chemical Industry Co., Ltd. Polyethylene glycol (molecular weight 6000) (hereinafter may be abbreviated as PEG in this example) was provided by NOF Corporation (Tokyo, Japan).

[0055] <Preparation of PEG-Cetilistat-containing emulsion> A PEG-Cetilistat-containing emulsion was prepared according to the following procedure. 1) Cetilistat (50 mg) and PEG (1500 mg) (both at room temperature, in powder form) were stirred and mixed at room temperature for 5 minutes. 2) The two components were stirred and mixed in a warm bath (75 °C) for 15 minutes. 3) Deionized distilled water (DDW) (3450 mg) was added to this mixture, and it was further stirred and mixed in a warm bath (75 °C) for 15 minutes. 4) 1 ml of the obtained emulsion was taken (1 ml = 1140 mg, RT), diluted 11.4 times (10.4 ml of DDW was added) to obtain a turbid emulsion of Cetilistat (concentration 1 mg / ml) (hereinafter may be abbreviated as PEG-C in this example).

[0056] <Examination of the protective effect of PEG-C on pancreatic acinar cells (in vitro)> (Cell collection) Primary acinar cells were collected according to the following procedure. 1) SD rats (9 - 12 weeks old, Shimizu Experimental Materials Co., Ltd.) were euthanized and exsanguinated. 2) A medium containing 2 mg / ml of collagenase D (Roche, derived from Clostridium histolyticum, 0.24 U / mg) was injected into the rat bile duct. 3) The pancreas was excised and incubated in 15 ml of the collagenase D-containing medium at 37 °C for approximately 20 minutes. 4) Subsequently, the solution was shaken for 20 seconds to obtain a cell suspension. 5) It was filtered through a 1 mm metal mesh, centrifuged at 100 x g for 3 minutes, and then the supernatant was discarded. 6) The precipitate was resuspended in the medium (50 ml) again, filtered through a cell strainer 352350 (Falcon) with a 70 μm mesh, centrifuged at 100 x g for 3 minutes, and the supernatant was discarded to recover the target cells.

[0057] (Cell seeding) Approximately 1.5×10 5 cells / ml were dispensed at 1 ml per well into each well of a 48-well plate (made of polystyrene, without surface treatment, Asahi Glass). Although it was difficult to accurately determine the cell density because the cells were separated into a mulberry-like state during cell collection, the cell suspension was dispensed so that each well had the same density for the experiment.

[0058] The composition of the medium (per 1000 ml) used in the above experiment is as follows. Krebs-Ringer bicarbonate buffer (K4002-1L, Sigma-Aldrich): 1 bottle Calcium chloride: 1 mM HEPES: 10 mM Sodium bicarbonate: 10 mM Bovine serum albumin (fatty acid-free) (08587-84, Nacalai Tesque): 1000 mg Trypsin inhibitor (soybean-derived) (202-09221, Fujifilm Wako Pure Chemical Industries, Ltd.): 100 mg

[0059] (Incorporation of neutral lipids and PEG-C into the medium) A stock solution (triolein concentration: 50 mM) was prepared by dissolving triolein (208-02981, Fujifilm Wako Pure Chemical Industries, Ltd.) in DMSO (13408-64, Nacalai Tesque). This triolein-DMSO stock solution was mixed with 0.5 ml of medium in which PEG-C had been dissolved to twice the final concentration, and 0.5 ml of the cell suspension to make 1 ml. This was then dispensed into wells to obtain the desired concentration.

[0060] (LDH assay) 100 μl of the supernatant was collected at each time point, and the absorbance was measured using an LDH Cytotoxicity Assay Kit-WST (Dojindo Laboratories, Ltd.) according to the manufacturer's instructions, to determine lactate dehydrogenase (LDH) activity.

[0061] <Animal experiments> (animal) All animal experiments were approved by the Institutional Animal Care and Use Committee of Kyoto University and were conducted in accordance with the ARRIVE guidelines. Obese rats were generated according to a previous report, as outlined below. Three-week-old male Sprague-Dawley rats (Shimizu Experimental Materials Co., Ltd.) were fed a high-fat diet (D12451, Research Diets Inc.) ad libitum for over six weeks. Rats weighing 400–450 g were used. Male animals were used to minimize variations in body weight and biochemistry. Rats were housed at a temperature of 21–25°C under a 12-hour light–dark cycle and had free access to water.

[0062] (Creation of a rat model of pancreatic fistula and adipose tissue damage treatment; see Figure 1) Pancreatic fistula model rats (PT group) were created by pancreatic resection according to a previous report. Rats were anesthetized with sevoflurane inhalation and intraperitoneal injection of pentobarbital. Analgesia was achieved with a subcutaneous injection of buprenorphine. After midline laparotomy, the pancreas was secured ventral to the portal vein without passive movement. The splenic artery and vein were preserved. The rat pancreatic duct consists of four parts: the gastric duct, duodenal duct, common duct, and splenic duct. The splenic duct and surrounding pancreatic parenchyma were transected with scissors at the level directly above the portal vein. Bleeding from the cut end of the pancreas was stopped by applying natural pressure with gauze. Care was taken not to damage the adipose tissue. To clarify the effects of adipose tissue injury, some animals underwent adipose tissue injury (the treatment was coded as +F). Bilateral epididymal adipose tissue was dissected, placed in a 35-mm sterile polystyrene dish, and immersed in 1 ml of saline. Adipose tissue was coagulated for 5 minutes using a bipolar electrocautery system (ConmedSaver Genesis) (power output 50 W). After cooling at room temperature for 3 minutes (to avoid burns), the injured adipose tissue and exudate were returned to the abdominal cavity (PT+F group).

[0063] (Animal grouping and treatment) The experimental animals were divided into groups, and each group was given the following treatments. Sham group: Only a single laparotomy was performed. Sham+F group: Single laparotomy and fat injury procedures were performed. PT group: Only pancreatic resection was performed. PT+F group: Pancreatic resection and fat injury procedures were performed. PT+F+C: After pancreatic resection, PEG-C was administered (PEG-C; 1 ml intraperitoneal administration), followed by fat injury treatment. PT+F+V: After pancreatic resection, an aqueous PEG solution was administered, followed by fat injury treatment (aqueous PEG solution; 1 ml intraperitoneal administration; PEG content was the same as that in PEG-C).

[0064] (Blood sample collection) Rats were sacrificed 24 hours after surgery or observed for 7 days for survival analysis. Blood samples were collected from the inferior vena cava immediately after death, and serum was separated. Peritoneal lavage fluid was collected from the peritoneal cavity after irrigation with 4 ml of saline, centrifuged, and the supernatant was used as ascites. All ascites and serum samples were analyzed immediately or frozen at -80°C for later analysis and used only after thawing once.

[0065] (Biochemical Assays) Amylase and lipase activities were performed at a specialized clinical laboratory (Japan Medical Clinical Laboratory Co., Ltd.).

[0066] (Measurement of free fatty acid concentration) The free fatty acid concentration of each blood sample was determined by high-performance liquid chromatography (HPLC) using a labeling reagent (XSRFAR01; YMC Co., Ltd.) (detector: Prominence SPD-20A, Shimadzu; column: YMC Pack FA, YMC Co., Ltd.). Margaric acid (C17:0) was used as an internal standard. Total free fatty acid (TFA) concentrations were calculated by adding individual C12:0, C14:0, C16:0, C18:0, C16:1, C18:1, and C18:2 fatty acids. Unsaturated fatty acid concentrations were calculated by adding individual C16:1, C18:1, and C18:2 fatty acids, according to a previous report.

[0067] (Histological analysis) After sacrifice, tissues were removed and fixed in 10% buffered formalin for 48 hours, and 4-μm-thick paraffin-embedded sections were prepared. Hematoxylin and eosin staining and Alizarin Red S staining (Sigma-Aldrich) for the detection of calcium deposits were performed according to standard protocols.

[0068] (statistical analysis) After applying Levene's test of variance, groups were compared using t-tests, Mann-Whitney U tests, one-way ANOVA with Tukey's multiple comparison test, or Steel-Dwass tests, as appropriate. Categorical variables were compared using Fisher's exact test. Experimental results are shown as mean values. Five independent animals / samples were analyzed unless otherwise stated. Survival analysis was performed using Kaplan-Meier plots, assessed by the log-rank test. Data analysis was performed using JMP Pro version 14.0 (SAS).

[0069] Example 1: Evaluation of the effect of fat damage on pancreatic fistula model mice

[0070] As described above, the generation of pancreatic fistula model mice (pancreatic resection procedure) and fat injury procedure were performed according to the schematic diagram shown in FIG. (1) In the PT group, a mild pancreatic fistula-like model, no significant changes were observed in the abdominal cavity compared to the sham group, and no significant changes were observed even after fat damage treatment (+F) in the sham group (Sham+F group). On the other hand, in the PT+F group, which combined both treatments, significant white deposits were observed in the abdominal cavity (Figure 2).

[0071] (2) The free fatty acid concentration (μM) in the ascites of each group was examined. In the PT+F group, it was found that high levels of lipolysis occurred in the abdominal cavity, resulting in the production of free fatty acids. Pancreatic resection alone or fat injury alone did not significantly increase the production of free fatty acids compared to the control group (Figure 3).

[0072] (3) Pancreatic enzyme activity (IU / L) in ascites was examined. In the PT group, significant pancreatic juice leakage occurred in the abdominal cavity. In the PT+F group, significantly more pancreatic juice leakage was observed than in the PT group, and significant increases in pancreatic enzyme activity in the ascites were observed (Figure 4).

[0073] (4) Histological findings of the pancreas were examined. In the PT group, mild edema and inflammatory cell infiltration were observed in the pancreas. Similar changes were observed in the PT+F group, but in the PT+F group, hyaline deposits were observed on the surface of the pancreas, surrounded by inflammatory cell infiltration. These deposits stained orange with Alizarin S staining and were thought to be deposits of fatty acid calcium (Figure 5).

[0074] (5) The survival rates of the PT group and the PT+F group were examined. No deaths were observed in the PT group during the one-week observation period. The PT+F group showed a significantly decreased survival rate, indicating more severe disease (Figure 6).

[0075] [Example 2: Evaluation of the effect of lipase inhibitors (in vitro)]

[0076] (1) Pancreatic acinar cells produce and secrete pancreatic enzymes, including lipase, and in this environment, triglycerides are broken down into free fatty acids. To examine the effect of triglycerides on pancreatic acinar cells, we administered triglyceride (triolein) to the culture medium and examined its cytotoxicity. The LDH concentration in the supernatant increased over time depending on the triglyceride concentration in the medium, confirming the time-dependent enhancement of cytotoxicity (Figure 7). This change was characteristic of pancreatic acinar cells and was not observed at all in similar experiments with other types of cells (rat MSCs (mesenchymal stem cells)).

[0077] (2) It was confirmed that the administration of PEG-C into the culture medium reduced damage to pancreatic acinar cells even in the presence of neutral fat. As shown in Figure 8, the PEG-C group demonstrated a dose-dependent protective effect on pancreatic acinar cells compared to the control group (no PEG-C added). In particular, a significant decrease in the oleic acid concentration in the medium was observed in the 100 μM PEG-C group.

[0078] [Example 3: Evaluation of the effect of lipase inhibitors (in vivo)]

[0079] (1) The therapeutic effect of PEG-C on a mouse model of pancreatic fistula was evaluated (intraperitoneal findings). When examining changes in the abdominal cavity, the white deposits observed in the PT+F group were suppressed by administration of PEG-C (PT+F+C group). Administration of vehicle alone (aqueous PEG solution) resulted in changes similar to those observed in the PT+F group (PT+F+V group) (Figure 9).

[0080] (2) Pancreatic enzyme activity (IU / L) in ascites was examined. Compared with the PT+F group, the PT+F+C group showed a significant reduction in pancreatic juice leakage, whereas the vehicle-administered group (PT+F+V group) did not show this effect (Figure 10).

[0081] (3) When the free fatty acid concentration (μM) in ascites was examined, it was found that the free fatty acid concentration in ascites was significantly lower in the PT+F+C group than in the PT+F group, indicating that free fatty acid production was significantly suppressed (Figure 11).

[0082] (4) The survival rate of each group was examined. No deaths were observed in the PT+F+C group during the observation period, confirming that the survival rate was significantly improved compared to the PT+F group (FIG. 12).

[0083] (summary) The results of the verification from various viewpoints shown in Examples 1 to 3 revealed that intraperitoneal administration of PEG-C can suppress the aggravation of pancreatic fistula by suppressing intraperitoneal lipolysis. Lipase inhibitors are an effective strategy for preventing and treating pancreatic fistula.

[0084] Example 4: Preparation of cetilistat-hydrogel complex <Hydrogel-forming ingredients> The hydrogel-forming components used were commercially available tetrafunctional polyethylene glycol modifying agents, SUNBRIGHT PTE-100GS (molecular weight 10,000, terminal succinimidyl glutarate) (NOF Corporation) and SUNBRIGHT PTE-100SH (molecular weight 10,000, terminal thiol) (NOF Corporation). <Preparation of PEGgel Adjusting Solution> A solution (pH 6.0) was prepared by dissolving SUNBRIGHT PTE-100GS in dilute hydrochloric acid to a concentration of 20% (w / v) (Solution 1). On the other hand, a solution (pH 9.6) was prepared by dissolving SUNBRIGHT PTE-100SH in a phosphate-carbonate buffer to a concentration of 20% (w / v) (Solution 2). Solutions 1 and 2 were placed in a Nordson DIY dual syringe 4 ml, and a mixing needle outlet was attached to obtain a PEGgel adjusting solution. By extruding each solution with this dual syringe, a hydrogel is rapidly formed (hereinafter may be abbreviated as PEGgel in this example).

[0085] <Preparation of Cetylistat-Hydrogel Composite Adjusting Solution> Cetylistat and SUNBRIGHT PTE-100GS were mixed in a powder state at a ratio of 1:50 (weight ratio), stirred in a warm bath at 75°C for 5 minutes, and then dilute hydrochloric acid was added so that the concentration of SUNBRIGHT PTE-100GS became 20% (w / v) to prepare a turbid emulsion-like solution 3 (pH 6.0). On the other hand, SUNBRIGHT PTE-100SH was added to a phosphate-carbonate buffer solution to a concentration of 20% (w / v) to prepare Solution 4 (pH 9.6). Solutions 3 and 4 were placed in a Nordson DIY dual syringe 4 ml, and a mixing needle outlet was attached to obtain a cetylistat-hydrogel composite adjusting solution. By extruding each solution with this dual syringe, a hydrogel containing cetylistat is rapidly formed (hereinafter may be abbreviated as PEGgel-C in this example). In this hydrogel, the cetylistat concentration was 2.5 mg / ml (equivalent to about 1; 2.5 mg / g specific gravity), and when 500 μl of the hydrogel was administered around the pancreas of an animal, the cetylistat administration amount was 1.25 mg.

[0086] [Example 5: Evaluation of the Effect of PEGgel-C (in vivo)] To evaluate the effect of PEGgel-C on pancreatic fistula, animal experiments were conducted using a pancreatic fistula model mouse. <Experimental Method> The experimental animals used were the same as those described in the above section on <Animal Experiments> (Animals). The animal experiments were carried out according to the following procedure. 1. On the day of the experiment, PEGgel preparation solution and cetilistat-hydrogel complex preparation solution were prepared. 2. Under general anesthesia, the pancreas of the rat was excised, and after hemostasis (similar to the PT group in the animal experiment), 500 μl of PEGgel preparation or cetilistat-hydrogel complex preparation was administered around the pancreas (PEGgel (N=2), PEGgel-C (N=3), PEGgel-C group: cetilistat dose 1.25 mg / body). 3. Completion of gelation after administration of the drug solution was confirmed (approximately 1 minute after administration). 4. The fat injury procedure shown in Figure 1 was performed, and the fat was cauterized and coagulated, and the exudate was returned to the abdominal cavity (+F procedure), after which the wound was closed. 5. 24 hours after surgery, the rats were sacrificed and tissues were removed for histological analysis.

[0087] The animals in this animal experiment were divided into the following groups: PEGgel-C group: A group administered PEGgel-C in the above animal experiment (N=3). PEGgel group: A group administered PEGgel in the above animal experiment (N=2). PT group: A group that underwent only pancreatic resection (see the data obtained in the animal experiment conducted in Example 1) (N=5). PT+F group: a non-treatment group to which neither PEGgel-C nor PEGgel was administered (see the data obtained in the animal experiment conducted in Example 1) (N=5).

[0088] <Experimental Results> (1) Histological findings (PEGgel-C administration group) As shown in Figures 13 and 14, in the PEGgel-C group, a clear reduction (inhibition) was confirmed in all evaluation items, including general inflammation and saponification in the abdominal cavity, compared to the untreated group (see Figure 17). (PEGgel administration group) As shown in Figures 15 and 16, no obvious changes were observed in the overall findings within the abdominal cavity in the PEGgel group compared to the untreated group (see Figure 17).

[0089] (2) Examination of pancreatic enzyme activity (IU / L) in ascites The measurement results are shown in Figure 18. It was confirmed that the leakage of pancreatic enzymes into the ascites was significantly suppressed in the PEGgel-C group compared to the untreated group (PT+F group). For the PT group and PT+F group, the data obtained in Example 1 above was used.

[0090] (summary) It was confirmed that covering the pancreas with a cetilistat-hydrogel complex (PEGgel-C) may be able to prevent the progression of pancreatic fistula. The hydrogel formulation containing lipase inhibitors offers a new therapeutic strategy for pancreatic fistula by efficiently distributing the lipase inhibitor locally in the pancreas.

[0091] Example 6: Preparation of orlistat-hydrogel complex An orlistat-hydrogel complex preparation solution containing orlistat was obtained by the same procedure as in Example 4 <Preparation of cetilistat-hydrogel complex preparation solution>, except that instead of mixing cetilistat and SUNBRIGHT PTE-100GS in a powdered state at a weight ratio of 1:50, orlistat and SUNBRIGHT PTE-100GS in a powdered state at a weight ratio of 1:30. By pushing out each solution using the dual syringe, a hydrogel containing orlistat was quickly formed (hereinafter, sometimes abbreviated as PEGgel-O in this example). In this hydrogel, the orlistat concentration was 3.3 mg / ml (specific gravity approximately 1; equivalent to 3.3 mg / g), and the amount of orlistat administered when 500 μl of the hydrogel was administered around the pancreas of an animal was 1.67 mg.

[0092] Example 7: Evaluation of the effect of PEGgel-O (in vivo) The same procedure as in Example 5 was carried out except that PEGgel-O was used instead of PEGgel-C, and the effect on pancreatic fistula was evaluated.

[0093] The animals in this animal experiment were divided into the following groups: PEGgel-O group: The group administered PEGgel-O in the above animal experiment (N=2).

[0094] <Experimental Results> (1) Histological findings (PEGgel-O administration group) As shown in Figure 19, the PEGgel-O group showed a clear reduction (inhibition) in both the overall inflammatory findings and saponification in the abdominal cavity compared to the untreated group (see Figure 17).

[0095] (2) Examination of pancreatic enzyme activity (IU / L) in ascites Ascites amylase activity was measured as a measure of pancreatic enzyme activity, and the result was 3559 IU / L in the PEGgel-O group, confirming that leakage of pancreatic enzymes into the ascites was significantly suppressed compared to the untreated group (PT+F group). For the PT and PT+F groups, the data (16868 IU / L) obtained in Example 1 above was used.

[0096] (summary) It was confirmed that coating the pancreas with an orlistat-hydrogel complex (PEGgel-O) may be able to prevent the progression of pancreatic fistula.

[0097] Example 8: Preparation of cetilistat-Bolheal (fibrin gel) complex <Hydrogel-forming components> As a hydrogel-forming component, Teijin Pharma Co., Ltd.'s Bolheal tissue adhesive 1ml preparation 注1 (hereinafter sometimes abbreviated as Bolheal) was used. Note 1) About Bolheal Tissue Adhesive The Bolheal tissue adhesive formulation consists of freeze-dried fibrinogen powder, fibrinogen solution, freeze-dried thrombin powder, and thrombin solution, and the details are as follows. Vial 1: Fibrinogen lyophilized powder (Contains fibrinogen and blood clotting factor XIII derived from human plasma) Vial 2: Fibrinogen Solution (Contains aprotinin derived from bovine lung) Vial 3: Thrombin lyophilized powder (Contains thrombin derived from human plasma) Vial 4: Thrombin Solution (Contains calcium chloride hydrate) By using Bolheal tissue adhesive formulation as a hydrogel-forming component, thrombin acts on fibrinogen, a glycoprotein in plasma, to generate fibrin monomers, which are then polymerized by the action of calcium to form fibrin polymers, and the action of factor XIII crosslinks the fibrin polymers to form a mesh-like fiber called stabilized fibrin, producing the so-called fibrin gel.

[0098] <Preparation of cetilistat-Bolheal complex solution> A thrombin solution containing PEG-C was prepared by mixing 0.12 mL of the PEG-C prepared in the above example with 0.88 mL of Bolheal's thrombin solution. This solution was then added to the lyophilized thrombin powder to prepare a thrombin solution containing PEG-C. On the other hand, fibrinogen solution was added to freeze-dried fibrinogen powder according to the method described in the package insert for "Bolheal Tissue Adhesive 1ml Preparation" to prepare a fibrinogen solution. It was confirmed that a hydrogel containing cetilistat was rapidly formed by adding the thrombin solution containing the above-mentioned PEG-C and fibrinogen solution to the dual syringe attached to Bolheal and then pushing out each solution using this dual syringe (hereinafter, this may be abbreviated as Bolheal-C in this example). In this hydrogel, the cetilistat concentration was 2.0 mg / ml (specific gravity approximately 1; equivalent to 2.0 mg / g), and the amount of cetilistat administered when 500 μl of the hydrogel was administered around the pancreas of an animal was 1.0 mg.

[0099] [Example 9: Evaluation of the effect of Bolheal-C (in vivo)] The same procedure as in Example 5 was carried out except that Bolheal-C was used instead of PEGgel-C, and the effect on pancreatic fistula was evaluated.

[0100] The animals in this animal experiment were divided into the following groups: Bolheal-C group: The group administered Bolheal-C in the above animal experiment (N=2).

[0101] <Experimental Results> (1) Histological findings (Bolheal-C administration group) As shown in Figure 20, in the Bolheal-C group, a reduction (suppression) was confirmed in both evaluation items of general inflammation and saponification in the abdominal cavity compared to the untreated group (see Figure 17).

[0102] (summary) It has been confirmed that covering the pancreas with the cetilistat-Bolheal complex (Bolheal-C) may prevent the progression of pancreatic fistula.

[0103] Comparative Example 1: Preparation of Bolheal alone A hydrogel consisting of Bolheal alone, without cetilistat, was prepared by the same procedure as in Example 8 <Preparation of cetilistat-Bolheal complex preparation>, except that Bolheal thrombin solution was used instead of the PEG-C-containing thrombin solution. (Hereinafter, in this example, this may be abbreviated as Bolheal alone.)

[0104] [Comparative Example 2: Evaluation of the effect of Bolheal alone (in vivo)] The same procedure as in Example 5 was carried out except that Bolheal alone was used instead of PEGgel-C, and the effect on pancreatic fistula was evaluated.

[0105] The animals in this animal experiment were divided into the following groups: Bolheal alone group: A group administered Bolheal alone in the above animal experiment (N=2).

[0106] <Experimental Results> (1) Histological findings (Bolheal monotherapy group) As shown in FIG. 21, in the Bolheal alone group, inflammation and saponification were observed throughout the abdominal cavity, similar to the untreated group (see FIG. 17).

[0107] (summary) Lipase inhibitors have been shown to be effective preventive and therapeutic strategies for pancreatic fistula, and hydrogel formulations of lipase inhibitors have also been shown to be an effective method of administration. [Industrial Applicability]

[0108] The present invention provides an agent for preventing or treating pancreatic leakage and / or pancreatic fistula after abdominal surgery, and a method for preventing or treating pancreatic leakage or pancreatic fistula after abdominal surgery, and is useful in the field of medicine.

[0109] This application is based on patent application No. 2020-18309 (filing date: February 5, 2020) and patent application No. 2020-167881 (filing date: October 2, 2020) filed in Japan, the contents of which are incorporated in their entirety herein.

Claims

1. Contains a lipase inhibitor as an active ingredient, the lipase inhibitor is a combination of one or two drugs selected from orlistat and cetilistat; A hydrogel preparation containing a complex of a lipase inhibitor and a hydrogel.

1. A preventive or therapeutic agent for pancreatic leakage and / or pancreatic fistula after abdominal surgery.

2. The preventive or therapeutic agent according to claim 1 for preventing or treating pancreatic fistula after abdominal surgery.

3. The preventive or therapeutic agent according to claim 2, wherein the pancreatic fistula is a clinically relevant postoperative pancreatic fistula (CR-POPF).

4. The preventive or therapeutic agent according to any one of claims 1 to 3, wherein the abdominal surgery is pancreatic surgery.

5. The preventive or therapeutic agent according to any one of claims 1 to 4, characterized in that it contains 0.01 to 20% (w / w) of a lipase inhibitor.

6. The preventive or therapeutic agent according to any one of claims 1 to 5, wherein the hydrogel is a hydrogel preparation formed from any one of the following combinations of hydrogel-forming components: 1) A combination of a hydrophilic biocompatible polymer and a biocompatible hydrophilic polymer crosslinker; 2) A combination of a synthetic biocompatible polymer having a functional group (polymer A) and a synthetic biocompatible polymer having a functional group that reacts with polymer A under in vivo conditions (polymer B); or 3) Combination of functionalized small molecule compounds and biocompatible hydrophilic polymeric crosslinkers.

7. The preventive or therapeutic agent according to claim 6, wherein the synthetic biocompatible polymer contains a polyalkylene oxide having a functional group.

8. The preventive or therapeutic agent according to claim 6, wherein the hydrophilic biocompatible polymer is at least one selected from gelatin, collagen, fibrinogen, fibrin, cellulose, chitosan, and derivatives thereof.

9. The preventive or therapeutic agent according to claim 6, wherein the hydrophilic biocompatible polymer is at least one selected from the group consisting of cross-linked gelatin, fibrinogen, fibrin, and derivatives thereof.

10. The preventive or therapeutic agent according to any one of claims 1 to 9, which contains 0.01 to 10% (w / w) of a lipase inhibitor.

11. The preventive or therapeutic agent according to claim 10, which contains a lipase inhibitor in an amount of 0.01 to 5% (weight of lipase inhibitor / weight of hydrogel-forming component).

12. The preventive or therapeutic agent according to any one of claims 1 to 11, further comprising a dispersing agent.

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