WO2017082121A1 - 徐放性局所投与剤 - Google Patents
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- WO2017082121A1 WO2017082121A1 PCT/JP2016/082456 JP2016082456W WO2017082121A1 WO 2017082121 A1 WO2017082121 A1 WO 2017082121A1 JP 2016082456 W JP2016082456 W JP 2016082456W WO 2017082121 A1 WO2017082121 A1 WO 2017082121A1
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
- A61K31/445—Non condensed piperidines, e.g. piperocaine
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0014—Skin, i.e. galenical aspects of topical compositions
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/06—Ointments; Bases therefor; Other semi-solid forms, e.g. creams, sticks, gels
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P23/00—Anaesthetics
- A61P23/02—Local anaesthetics
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/02—Drugs for disorders of the nervous system for peripheral neuropathies
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/10—Antimycotics
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P5/00—Drugs for disorders of the endocrine system
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/02—Inorganic compounds
Definitions
- the present invention relates to a sustained-release local administration agent, in other words, a sustained-release drug for local administration.
- a local anesthetic containing a local anesthetic can be considered.
- Local anesthetics are commonly used to suppress nociceptive pain and are usually administered by local injection.
- Pharmaceutical compositions for local injection usually contain a local anesthetic at a concentration of 0.2-2%.
- epidural anesthesia which has been considered the gold standard so far, has become the first choice for postoperative analgesia due to changes in the balance of risk and benefit due to standardization of postoperative anticoagulation therapy and minimally invasive surgery. I know it ’s a difficult situation.
- opioid IV-PCA As an alternative to epidural anesthesia, opioid IV-PCA, peripheral nerve block, and their combination are currently mainly used.
- the postoperative analgesic effect is required for about 1 to 3 days, whereas the duration of the effect of the peripheral nerve block by the injection of the local anesthetic is as short as about half a day.
- Patent Document 1 (Special Tables 2011-508788, WO2009-129149) reduces, prevents or treats pain and / or inflammation in a patient in need thereof.
- a plurality of implantable drug depots useful for the treatment of a first set of agents capable of releasing therapeutically effective bolus amounts of analgesics and / or anti-inflammatory drugs or pharmaceutically acceptable salts thereof at sites under the skin Including one or more drug depots and a second set of one or more drug depots capable of releasing a therapeutically effective amount of an analgesic and / or anti-inflammatory drug or pharmaceutically acceptable salt thereof over a period of at least 3 days.
- a plurality of implantable drug depots are disclosed.
- Patent Document 2 discloses a thermal gelation-stabilized pharmaceutical composition of at least one local anesthetic having a pH close to pKa of the local anesthetic.
- A one or more local anesthetics of the amide type (ATC code: N01BB),
- B 10-30% by weight of polyoxyethylene castor oil, and
- a pharmaceutical composition comprising at least 15% by weight of one or more surfactants to impart properties.
- Patent Document 3 Japanese Patent Publication No. 2013-523694, WO2011-121082, US Publication 2013-079371 describes a stabilized bioadhesive gelled aqueous pharmaceutical composition comprising (a) an anesthetic effective amount of 1 One or more local anesthetics; (b) monoglyceride or diglyceride in an amount of 15 to 70% by weight, or a mixture of its long chain fatty acids; and (c) free long chain saturated or unsaturated fatty acid in an amount of 5 to 60% by weight And a pharmaceutical composition having an anisotropic organic phase behavior that allows swelling at the site of administration containing excess moisture.
- Patent Document 4 Japanese Translation of PCT International Publication No. 2006-523731, WO 2004-92223 describes a crosslinked polysaccharide gel (epoxy crosslinked polysaccharide) useful for pharmaceutical use with improved wrinkle degradation characteristics (improved resistance to gel degradation). ).
- Paragraph No. 0043 of the same document discloses that it can be combined with an anesthetic.
- Patent Document 5 Japanese Translation of PCT International Publication No. 2007-521225, WO2005-009408 proposes a sustained-release preparation that releases an anesthetic such as bupivacaine over a short period of time, which is used for postoperative wound pain relief.
- Patent Document 5 discloses one containing an anesthetic dissolved or dispersed in a short-term gel excipient. And the polysaccharide containing an ester terminal group is illustrated as a gel excipient
- the anesthetic of Patent Document 5 is a viscous gel formed from a polymer and a solvent.
- Patent Document 6 International Publication WO 2005-087289, USP 7737214.
- Patent Document 6 has at least one active ester group introduced into a polysaccharide side chain and capable of reacting with an active hydrogen-containing group, and the active ester group and active hydrogen are brought into contact with water under alkaline conditions.
- An anti-adhesion material comprising a crosslinkable polysaccharide derivative capable of forming a cross-linked product by a covalent bond with a containing group has been proposed.
- Patent Documents 1 to 5 cannot achieve gel disappearance and drug release at the same time. Further, in Patent Documents 2 and 3, the release amount is linear with respect to time, the blood concentration rises quickly, and it is not a preferable profile that maintains a constant concentration for a certain period. Furthermore, in the anesthetic disclosed in Patent Document 2, 3 or 5, the solvent is immiscible with water. Such water-immiscible solvents are often toxic per se and are not desirable for the surgical site.
- the adhesion preventing material of Patent Document 6 has durability that exists for a certain period of time, and is effective as an adhesion preventing material. However, this anti-adhesion material does not contain a drug and does not have sustained drug release properties.
- the object of the present invention is to rapidly increase the blood concentration of a compound having a pharmacological action after topical application, to maintain a constant concentration for a certain period of time, and to release the compound continuously.
- a sustained-release topical preparation in which the disappearance of the gel ends substantially simultaneously.
- a sustained-release topical agent used after mixing wherein the sustained-release topical agent comprises a compound having a pharmacological action and an active ester group introduced into a polysaccharide side chain and capable of reacting with an active hydrogen-containing group
- a crosslinkable polysaccharide derivative capable of forming a cross-linked product by a covalent bond between the active ester group and the active hydrogen-containing group, and in a non-mixed state with the crosslinkable polysaccharide derivative, and at the time of mixing
- a pH adjuster for adjusting the pH to form a biodegradable gel containing the compound, and the sustained-release topical agent forms the biodegradable gel at the time of mixing.
- a sustained-release topical preparation in which the biodegradable gel formed has a pH of 5 or more.
- FIG. 1 is a graph showing experimental results using the sustained-release topical administration agent of the present invention.
- FIG. 2 is a graph showing experimental results using the sustained-release topical administration agent of the present invention.
- the sustained-release topical preparation of the present invention will be described with reference to examples.
- the sustained release topical administration agent of the present invention is a sustained release topical administration agent used after mixing.
- the sustained-release topical administration agent has a compound having a pharmacological action and at least one active ester group introduced into a polysaccharide side chain and capable of reacting with an active hydrogen-containing group, and contains an active ester group and an active hydrogen.
- the sustained-release topical preparation forms a biodegradable gel containing a compound having a pharmacological action when mixed, and the biodegradable gel formed has a pH of 5 or more.
- the crosslinkable polysaccharide derivative used in the sustained release topical administration agent of the present invention will be described.
- the crosslinkable polysaccharide derivative has at least one active ester group introduced into the polysaccharide side chain and capable of reacting with an active hydrogen-containing group.
- the polysaccharide (raw material) into which this active ester group is introduced will be described later, since the polysaccharide molecule inherently has a hydroxyl group, that is, has an active hydrogen-containing group, the polysaccharide in which the active ester group has been introduced into the polysaccharide.
- the derivative has an active ester group and an active hydrogen-containing group in one molecular chain, and exhibits self-crosslinking property under reaction conditions.
- This self-crosslinking property means that an active ester group and an active hydrogen-containing group react within one molecule or between molecules of a polysaccharide derivative to form a covalent bond. Further, when the active hydrogen-containing group on the surface of the living body is used for the reaction, this crosslinkable polysaccharide derivative exhibits adhesion to the surface of the living body.
- crosslinkable polysaccharide derivative may be referred to as an active esterified polysaccharide, and hereinafter, it may be simply referred to as a polysaccharide derivative.
- active esterified polysaccharide such as a polysaccharide derivative.
- polysaccharide derivative such as a polysaccharide derivative.
- single molecular chain or “intramolecular” means one molecule in a range connected by a continuous bond through a covalent bond.
- the polysaccharide derivative according to the present invention is an active esterified polysaccharide and essentially retains the polysaccharide skeleton. Therefore, in the following, the polysaccharide derivative may be described in parallel with the polysaccharide active esterification method (polysaccharide derivative production method).
- the active ester group introduced into the polysaccharide may be any group that can react with the active hydrogen-containing group to form a covalent bond.
- Such an active ester group usually has a strong electrophilic group on the carbonyl carbon of a carboxy group or a methylcarboxy group that is self-contained by a polysaccharide molecule or introduced by acidification, as compared with a normal ester. It is a bonded group.
- the active ester group is represented by “—COOX”
- the electrophilic group forming the alcohol moiety “—OX” is preferably a group introduced from an N-hydroxyamine compound. This is because the N-hydroxyamine compound is a relatively inexpensive raw material, and therefore, it is easy to industrially introduce an active ester group.
- N-hydroxyamine compounds for forming “-OX” include N-hydroxysuccinimide, N-hydroxynorbornene-2,3-dicarboxylic acid imide, 2-hydroxyimino-2-cyanoacetic acid.
- Representative examples include ethyl ester, 2-hydroxyimino-2-cyanoacetamide, N-hydroxypiperidine and the like.
- the active ester group of the polysaccharide derivative may be used alone or in combination of two or more. Among such active ester groups, succinimide ester groups are preferred.
- the polysaccharide derivative used in the present invention has at least one active ester group in the molecule, but usually has two or more in one molecule in order to form a crosslinked matrix. Although depending on the purpose of use, it is preferably 0.1 to 2 mmol / g when expressed in terms of the amount of active ester groups per gram of the dry weight.
- the polysaccharide into which the active ester group is introduced and which constitutes the main skeleton of the polysaccharide derivative is not particularly limited as long as it has two or more monosaccharide structures in the main skeleton.
- Such polysaccharides are monosaccharides such as arabinose, ribose, xylose, glucose, mannose, galactose, fructose, sorbose, rhamnose, fucose, ribodeose; disaccharides such as trehalose, sucrose, maltose, cellobiose, gentiobiose, lactose, melibiose; Examples include those formed by covalently bonding polysaccharides of three or more sugars such as raffinose, gentianose, meretitol, and stachyose, and those having a functional group introduced thereto.
- such polysaccharides may be naturally occurring or artificially synthe
- the weight average molecular weight of the polysaccharide that is the main skeleton of the polysaccharide derivative is not particularly limited.
- it is a polysaccharide having a weight average molecular weight of 5,000 to 2.5 million corresponding to a combination of several tens to several thousand of the above monosaccharides, disaccharides, or trisaccharides. This is because with such a polysaccharide, it is easy to adjust the hardness of the gel after the polysaccharide derivative according to the present invention is crosslinked, and it is easy to introduce a plurality of active ester groups and active hydrogen-containing groups into one molecular chain. More preferably, it is a polysaccharide having a weight average molecular weight of 10,000 to 1,000,000.
- the raw material polysaccharide that forms the main skeleton of the polysaccharide derivative has the above-described constituents and has a carboxylic acid group for forming an active ester group “—COOX” in the active esterification precursor stage (hereinafter referred to as an acid group-containing polysaccharide).
- an acid group-containing polysaccharide (Sometimes called polysaccharides).
- the carboxylic acid group here refers to a carboxy group and / or a carboxyalkyl group (hereinafter sometimes referred to as a carboxylic acid group), and the carboxyalkyl group refers to a carboxymethyl group, a carboxyethyl group, a carboxypropyl group.
- a functional group in which a carboxy group is bonded to an alkyl skeleton as exemplified by carboxyisopropyl group, carboxybutyl group and the like.
- the raw material polysaccharide may be an acid group-containing polysaccharide at the precursor stage of the crosslinkable polysaccharide derivative, may be a natural polysaccharide that self-holds a carboxylic acid group, and is itself a polysaccharide that does not have a carboxylic acid group. It may be a polysaccharide introduced with a carboxy group and / or a carboxyalkyl group.
- carboxylic acid group-containing polysaccharides natural polysaccharides having a carboxy group, carboxylated polysaccharides introduced with a carboxy group, carboxymethylated polysaccharides introduced with a carboxymethyl group, and carboxyethylated polysaccharides introduced with a carboxyethyl group are preferred. . More preferred are natural polysaccharides having a carboxy group, carboxylated polysaccharides introduced with a carboxy group, and carboxymethylated polysaccharides introduced with a carboxymethyl group.
- the above-mentioned natural polysaccharide having the carboxylic acid group is not particularly limited, and examples thereof include pectin containing galacturonic acid and hyaluronic acid.
- pectin containing galacturonic acid and hyaluronic acid.
- “GENUE pectin” from CP Kelco (Denmark) can be used for pectin
- hyaluronic acid FCH” from Kibunsha (Japan) can be used as hyaluronic acid.
- Pectin is a polysaccharide mainly composed of galacturonic acid. About 75 to 80% or more of pectin is composed of galacturonic acid, and other components are mainly composed of other sugars.
- Pectin is a polysaccharide formed by binding galacturonic acid and other sugars at the above ratio.
- Hyaluronic acid is used as an ophthalmic surgical aid, a knee osteoarthritis therapeutic agent, and the like.
- Hyaluronic acid does not contain galacturonic acid.
- the carboxy group and / or carboxyalkyl group of the polysaccharide derivative is preferably “non-salt type” in which the salt is not coordinated, and the polysaccharide derivative finally obtained is preferably not in the salt form.
- the “salt” includes inorganic salts such as alkali metals and alkaline earth metals, quaternary amines such as tetrabutylammonium (TBA), and halogen salts such as chloromethylpyridylium iodide.
- TSA tetrabutylammonium
- halogen salts such as chloromethylpyridylium iodide.
- “Non-salt type” means that these “salts” are not coordinated, and “not in salt form” means that these salts are not included.
- the polysaccharide into which the carboxy group and / or carboxyalkyl group is introduced is not particularly limited, and examples thereof include dextran and pullulan.
- the above dextran is used as a plasma substitute.
- Dextran includes “Dextran T fractions” from Amersham Biosciences (Japan), and Pullulan includes “Pullulan PI-20” from Hayashibara (Japan). Pullulan is used as a pharmaceutical additive including oral drugs, and those with low biological contamination such as endotoxin are suitable.
- Any polysaccharide that is generally commercially available can be used in the present invention.
- the polysaccharide having a proven record in the above medical use is a polysaccharide that can be suitably used in terms of safety.
- the carboxylation reaction of the polysaccharide can be carried out without particular limitation using a known oxidation reaction.
- the type of the carboxylation reaction is not particularly limited, and examples thereof include dinitrogen tetroxide oxidation, fuming sulfuric acid oxidation, phosphoric acid oxidation, nitric acid oxidation, and hydrogen peroxide oxidation, each of which is a commonly known reaction using a reagent. Can be selected and oxidized.
- Each reaction condition can be appropriately set depending on the amount of carboxy group introduced.
- a polysaccharide for example, by suspending a polysaccharide as a raw material in chloroform or carbon tetrachloride and adding dinitrogen tetroxide, the hydroxyl group of the polysaccharide can be oxidized to prepare a carboxylated polysaccharide (polysaccharide carboxylate).
- the carboxyalkylation reaction can utilize a known polysaccharide carboxyalkylation reaction, and is not particularly limited. Specifically, in the case of carboxymethylation reaction, monochloroacetic acid is used after alkalizing the polysaccharide.
- the selected reaction can be selected. The reaction conditions can be appropriately set depending on the amount of carboxymethyl group introduced.
- any of the above-mentioned carboxylation or carboxyalkylation methods can be used, and the method is not particularly limited.
- Carboxyalkylation, particularly carboxymethylation, is preferred from the viewpoint of relatively easy control of the amount of introduction.
- the introduction of a carboxylic acid group is not particularly limited to introduction into a polysaccharide that does not itself have a carboxylic acid group.
- a carboxy group and / or a carboxymethyl group may be further introduced into a natural polysaccharide having its own carboxylic acid group, for example, the hyaluronic acid.
- the acid group-containing polysaccharide may be used alone or in combination of two or more kinds. May be.
- the acid group-containing polysaccharide used for the active esterification is usually 0.1 to 5 mmol / g, preferably 0.4, in terms of the amount of carboxylic acid groups per 1 g of dry weight (assuming that the group is one molecule). -3 mmol / g, more preferably 0.6-2 mmol / g.
- the ratio of the amount of the carboxylic acid group is less than 0.1 mmol / g, the number of active ester groups derived from the group and serving as a crosslinking point is often insufficient.
- the ratio of the amount of carboxylic acid groups is more than 5 mmol / g, the polysaccharide derivative (uncrosslinked) becomes difficult to dissolve in a solvent containing water.
- the active esterification method of the acid group-containing polysaccharide is not particularly limited.
- the acid group-containing polysaccharide is combined with an electrophilic group introducing agent in the presence of a dehydration condensing agent.
- Examples thereof include a method of reaction, a method of using an ester exchange reaction in which an active ester group is introduced into a polysaccharide from a compound having an active ester group.
- the former method is suitable for the present invention, and this method (also referred to as the method of the present invention) will be mainly described below.
- the acid group-containing polysaccharide is usually prepared in a solution of an aprotic polar solvent and subjected to the reaction. More specifically, the method includes a solution preparation step in which a polysaccharide having a carboxy group or a carboxyalkyl group is dissolved in an aprotic polar solvent, and an electrophilic group introducing agent and a dehydrating condensing agent are added to the solution. Examples include a method of performing a reaction step of esterifying a carboxy group or a carboxyalkyl group of a polysaccharide, and a method of performing a purification step and a drying step of the reaction product.
- the polysaccharide is added to the solvent and heated to 60 ° C. to 120 ° C., whereby the polysaccharide is dissolved in the aprotic polar solvent. Therefore, as the acid group-containing polysaccharide to be active esterified by this method, among the polysaccharides exemplified above, those soluble in an aprotic polar solvent at a temperature between 60 ° C. and 120 ° C. are preferably used.
- the polysaccharide used in the reaction for introducing the electrophilic group preferably has a carboxy group or a carboxymethyl group in an acid form from the viewpoint of solubility in an aprotic polar solvent.
- Acid type means that the cation group of the carboxy group or carboxymethyl group is a proton.
- a polysaccharide having an acid-type carboxy group is called an acid-type (raw material) polysaccharide.
- pectin which is a polysaccharide having a carboxy group
- acid-type pectin Carboxymethyldextran having an acid-type carboxymethyl group
- CM dextran acid-type carboxymethyl dextran
- the “acid type” is synonymous with the “non-salt type” in that the counter cation species is a proton and is not in a salt form.
- Aprotic polar solvent is a polar solvent having no proton capable of forming a hydrogen bond with a nucleophile having an electrically positive functional group.
- the aprotic polar solvent that can be used in the production method according to the present invention is not particularly limited, but dimethyl sulfoxide (DMSO), N, N-dimethylformamide, N-methyl-2-pyrrolidone, N, N-dimethylacetamide, 1 , 3-Dimethyl-2-imidazolidinone.
- Dimethyl sulfoxide can be suitably used because the solubility of the polysaccharide in the solvent is good.
- an electrophilic group-introducing agent and a dehydrating condensing agent are added to the acid-type polysaccharide solution, and the carboxy group and / or the carboxymethyl group of the polysaccharide are activated esterified.
- the reaction temperature for the active esterification is not particularly limited, but is preferably 0 ° C. to 70 ° C., more preferably 20 ° C. to 40 ° C. While the reaction time varies depending on the reaction temperature, it is generally 1 to 48 hours, preferably 12 to 24 hours.
- Electrophilic group-introducing agent refers to a reagent that introduces an electrophilic group into a carboxy group or carboxyalkyl group and converts them into an active ester group.
- the electrophilic group-introducing agent is not particularly limited, but an active ester-inducing compound that is widely used for peptide synthesis can be used, and an example thereof includes an N-hydroxyamine-based active ester-inducing compound.
- the N-hydroxyamine-based active ester-derived compound is not particularly limited.
- N-hydroxysuccinimide N-hydroxynorbornene-2,3-dicarboxylic acid imide, 2-hydroxyimino-2-cyanoacetic acid ethyl ester
- Examples include 2-hydroxyimino-2-cyanoacetamide, N-hydroxypiperidine and the like.
- N-hydroxysuccinimide is preferable because it has a track record in the field of peptide synthesis and is easily available commercially.
- the “dehydration condensing agent” is a condensation of a carboxy group or a carboxyalkyl group with an electrophilic group introducing agent when an electrophilic group introducing agent is used for the carboxy group or carboxyalkyl group to form an active ester group.
- One water molecule to be generated is extracted, that is, dehydrated, and both are ester-bonded.
- the dehydrating condensing agent is not particularly limited. For example, 1-ethyl-3-dimethylaminopropylcarbodiimide hydrochloride (EDC ⁇ HCl), 1-cyclohexyl- (2-morpholin-4-ethyl) -carbodiimide ⁇ meso-p- And toluene sulfonate.
- EDC ⁇ HCl 1-ethyl-3-dimethylaminopropylcarbodiimide hydrochloride
- the unreacted electrophilic group introducing agent, dehydrating condensing agent, and reaction by-products are removed from the reaction solution by means such as ordinary reprecipitation, filtration and / or washing.
- the polysaccharide derivative according to the present invention can be obtained.
- the drying step in order to remove the washing solvent from the polysaccharide derivative obtained in the purification step, it may be dried by a commonly used method.
- the amount of the active ester group of the polysaccharide derivative is preferably 0.1 to 2 mmol / g in the end, and in the above, the active substance group is obtained so that such a polysaccharide derivative is obtained.
- the amount of the active ester group introduced into the carboxy group of the esterified raw material polysaccharide can be controlled.
- the mixing amount of the electrophilic group introduction agent and the dehydration condensing agent can be adjusted in the reaction step.
- the ratio (Z / X) of the number of moles (Zmmol) of the dehydration condensing agent to the number of moles (Xmmol) of all carboxy groups of the polysaccharide is 0.1 ⁇ Z / X ⁇ 50 at the reaction temperature described above. It is preferable that the addition conditions satisfy the conditions.
- Z / X is less than 0.1, the reaction efficiency is low because the addition amount of the dehydrating condensing agent is small, making it difficult to achieve the desired active ester group introduction rate.
- Z / X is greater than 50, the dehydrating condensing agent This is because, although the amount of added is large, the introduction rate of the active ester group is high, but the obtained polysaccharide derivative is difficult to dissolve in water.
- the number of moles (Ymmol) of the electrophilic group-introducing agent relative to the number of moles (Xmmol) of all carboxy groups of the polysaccharide is not particularly limited as long as the reaction amount or more according to the introduction rate of the active ester group is added. It is preferable that the addition conditions satisfy 1 ⁇ Y / X ⁇ 100.
- the polysaccharide derivative according to the present invention usually has the hydroxyl group of the glucopyranose ring in the polysaccharide skeleton molecule, and thus possesses the active hydrogen-containing group by itself.
- the hydrogen-containing group is not limited to this, and may further have an active hydrogen-containing group introduced into the molecule as necessary.
- the active hydrogen-containing group possessed by the polysaccharide derivative may be one type or two or more types.
- the polysaccharide derivative according to the present invention can widely include functional groups such as known elements and atomic groups as long as the characteristics of the present invention are not impaired.
- functional groups include halogen elements such as fluorine, chlorine, bromine and iodine; carboxy groups; carboxyalkyl groups such as carboxymethyl groups, carboxyethyl groups, carboxypropyl groups, and carboxyisopropyl groups; silyl groups; An alkylene silyl group, an alkoxy silyl group, a phosphoric acid group etc. are mentioned.
- Such functional groups may be used alone or in combination of two or more.
- the introduction rate (%) of the active ester group is obtained with respect to the carboxy group-containing molar amount and the carboxymethyl group-containing molar amount (hereinafter referred to as total carboxy group (TC)) of the polysaccharide of the active esterification raw material.
- the ratio (AE / TC) of the molar amount (AE) of active ester groups in the polysaccharide derivative can be expressed by multiplying by 100.
- the active ester group introduction rate can be determined, for example, by the method described in Biochemistry Vol. 14, No. 7 (1975), p1535-1541. In particular, it may have a carboxy group and / or a carboxymethyl group of the remaining raw material polysaccharide when the active ester group is introduced at an introduction rate of the active ester group of less than 100%.
- Crosslinked structure means that the polysaccharide derivative according to the present invention forms a covalent bond within one molecular chain and / or between a plurality of molecular chains, and as a result, the molecular chain of the polysaccharide derivative takes a network-like three-dimensional structure. means.
- the active ester group and the active hydrogen-containing group can be bonded within one molecular chain, but may be cross-linked by covalent bonding between a plurality of molecules.
- the polysaccharide derivative according to the present invention which is water-soluble before the cross-linking reaction, forms a cross-linked structure as the reaction proceeds, the fluidity decreases, and becomes a water-insoluble mass (hydrous gel). Form.
- the polysaccharide derivative according to the present invention is defined as “self-crosslinking”, in which the property capable of forming a crosslinked structure by covalent bonding within its own molecular chain or between molecular chains without using any other crosslinking agent is defined. Is a self-crosslinking polysaccharide.
- the polysaccharide derivative according to the present invention is not only self-crosslinkable due to the involvement of an intramolecular active hydrogen-containing group as described above, but if the polysaccharide derivative is applied to a biological surface, the active hydrogen-containing group on the biological surface It is possible to show adhesiveness to the surface of a living body by the reaction of the active ester group. Such usage is a preferred embodiment of the polysaccharide derivative according to the present invention. Needless to say, self-crosslinking may occur simultaneously when applied to the surface of a living body.
- the active hydrogen-containing group involved in the reaction with the active ester group is not particularly limited as long as it is a group capable of reacting with the active ester group to form a covalent bond under the reaction conditions specific to the present invention. Also in this invention, it can apply to the thing illustrated as a general active hydrogen containing group. Specific examples include a hydroxyl group, an amino group, and a thiol group.
- the amino group includes a primary amino group and a secondary amino group.
- the active hydrogen-containing group is a hydroxyl group or a primary amino group, it is preferable because the reactivity with the active ester group is good and the time until crosslinking and gelation is short.
- the crosslinking reaction of the polysaccharide derivative is based on the formation of a covalent bond by the reaction between the active ester group and the active hydrogen-containing group.
- a method of adding a pH adjuster to the polysaccharide derivative solution to crosslink, and the like can be mentioned.
- the temperature is not particularly limited, but can be, for example, in the range of 10 ° C to 40 ° C.
- the polysaccharide derivative can be used in the form of powder or sheet. That is, the powdered polysaccharide derivative can be obtained by crushing or pulverizing the polysaccharide derivative obtained by the above-described synthesis reaction, and adjusting the particle size by adjusting the particle size if necessary. Although there is no particular limitation for reducing the particle size, freeze pulverization, mill pulverization and / or classification may be performed. After pulverization and pulverization, it can be adjusted to an arbitrary particle size distribution by sieving. The average particle size is not particularly limited, but an average particle size of several tens nm to several hundreds ⁇ m is preferable.
- the obtained powder can be prepared as a paste or aerosol by a commonly used method.
- the sheet-like polysaccharide derivative can be produced by a solution preparation step in which the polysaccharide derivative is dissolved in water and a drying step in which the solution is developed into a desired shape and dried by heating or freeze-drying.
- the sheet-like polysaccharide derivative can be obtained by preparing an aqueous solution in which the polysaccharide derivative is dissolved and freeze-drying.
- the pH of water for preparing the aqueous solution is preferably 3.0 to 7.5. When the pH is 3.0 or less, the resulting sheet exhibits strong acidity, and when it is 7.5 or more, the active ester group may be liberated.
- the heat-dried sheet can be obtained by spreading the aqueous solution on a substrate and heat-drying at 30 to 110 ° C. If necessary, it can also be heat-dried under reduced pressure.
- the freeze-dried sheet can be obtained by freezing the aqueous solution and drying it while freezing. If necessary, a normal freeze dryer can be used.
- the mixing timing of the pH adjuster (B) is not particularly limited, but is appropriately selected before or during use.
- the composition of the polysaccharide derivative (A) and the pH adjuster (B) may contain other substances as necessary, and the other substances may be mixed with or mixed with the polysaccharide derivative. It does not have to be.
- the pH adjuster (B) used in the present invention mainly comprises an aqueous solution, water for adjusting the pH of the biodegradable gel according to the present invention to 5.0 to 14, preferably 6.4 to 12. Or a salt (powder) or the like.
- the pH adjuster (B) is not particularly limited, and specifically, sodium carbonate aqueous solution or powder, sodium hydrogen carbonate aqueous solution or powder, phosphate buffer (disodium hydrogen phosphate-potassium dihydrogen phosphate), Examples include an acetic acid-ammonia buffer.
- sodium bicarbonate can be suitably used from the viewpoint of safety because a 7% aqueous solution (pH 8.3) thereof is used as an intravenous injection solution as a medical pH adjuster.
- a two-component system comprising an aqueous solution having a polysaccharide derivative concentration of 1 to 80% (W / V) and water adjusted to pH 7.5 to 10.5, which is maintained separately from the aqueous solution.
- both can be mixed at the time of use to obtain a mixed aqueous solution having a final polysaccharide derivative concentration of 0.1 to 60% (W / V).
- an aqueous solution having a polysaccharide derivative concentration of 1 to 80% (W / V) is mixed with the salt of the pH adjuster (B) added and dissolved to obtain a final polysaccharide derivative concentration.
- a normal mixing method can be selected for mixing, but it is preferable to perform the mixing until the mixed state becomes uniform, and it may be uniform so long as a desired reaction proceeds.
- a crosslinkable polysaccharide composition containing a polysaccharide derivative (A) and another polymer (C) can also be used.
- the polymer (C) is used to adjust the hardness and properties of the hydrated gel when the polysaccharide composition is crosslinked.
- One type of polysaccharide derivative (A) may be contained in the said polysaccharide composition, and 2 or more types may be contained.
- the polymer (C) is not particularly limited, but it is preferable to use a polymer (C) having two or more primary amino groups, thiol groups, or hydroxyl groups in one molecule.
- Specific examples of the polymer (C) include polyalkylene glycol derivatives, polypeptides, polysaccharides or derivatives thereof.
- the content of the polymer (C) in the polysaccharide composition according to the present invention is not particularly limited, but it is preferably 5 to 50% by mass with respect to the whole polysaccharide composition.
- the polymer (C) can be used alone or in combination of two or more.
- polyalkylene glycol derivatives examples include polyethylene glycol (PEG) derivatives, polypropylene glycol derivatives, polybutylene glycol derivatives, polypropylene glycol-polyethylene glycol block copolymer derivatives, and random copolymer derivatives.
- PEG polyethylene glycol
- polypropylene glycol derivatives examples include polypropylene glycol derivatives, polybutylene glycol derivatives, polypropylene glycol-polyethylene glycol block copolymer derivatives, and random copolymer derivatives.
- the basic polymer skeleton of the polyethylene glycol derivative include ethylene glycol, diglycerol, pentaerythritol, and hexaglycerol.
- the molecular weight of the polyalkylene glycol derivative is preferably 100 to 50,000. More preferably, it is 1,000 to 20,000.
- the polyethylene glycol derivative is not particularly limited.
- an ethylene glycol type polyethylene glycol derivative having a thiol group at both ends and a weight average molecular weight of 1,000, 2,000, 6,000 or 10,000 Ethylene glycol type polyethylene glycol derivatives having an amino group having a weight average molecular weight of 1,000, 2,000, 6,000 or 10,000, and a weight average molecular weight having a thiol group at three terminals of 5,000 or 10,000 Trimethylolethane type polyethylene glycol derivatives having 3 amino groups at the ends and a weight average molecular weight of 5,000 or 10,000, and 5 weight average molecular weights having thiol groups at the 4 ends.
- diglycerol type polyethylene glycol derivatives 4 diglycerol type polyethylene glycol derivatives having amino groups at the ends and weight average molecular weights of 5,000, 10,000 or 20,000, Pentaerythritol type polyethylene glycol derivative having a thiol group at the terminal and a weight average molecular weight of 10,000 or 20,000, and a pentaerythritol type polyethylene glycol having an amino group at the four terminals and a weight average molecular weight of 10,000 or 20,000 A derivative, a hexaglycerol type polyethylene glycol derivative having a thiol group at eight ends and a weight average molecular weight of 10,000 or 20,000, a weight average molecular weight having an amino group at eight ends of 10,000 or Hexaglycerol polyethylene glycol derivative of 0,000 are exemplified.
- Weight-average molecular weight is one of the numerical values representing the average molecular weight of a polymer. Since a polymer is a mixture of molecules having the same basic structural unit and different molecular lengths (chain lengths), it has a molecular weight distribution corresponding to the difference in molecular chain length. Average molecular weight is used to indicate its molecular weight. The average molecular weight includes a weight average molecular weight, a number average molecular weight, and the like. Here, the weight average molecular weight is used. In addition, the value (100%) of the weight average molecular weight in the present invention includes those having an upper limit of 110% and a lower limit of 90%.
- Polyethylene glycol derivatives can be prepared, for example, according to the method described in Chapter 22 of Poly (ethylene Glycol) Chemistry: Biotechnical and Biomedical Applications, J Milton Harris, Plenum Press, NY (1992). Alternatively, it can be chemically modified to contain multiple primary amino groups or thiol groups. Further, it can be purchased from Nippon Oil & Fats Co., Ltd. as a polyethylene glycol derivative (Sunbright HGEO-20TEA, Sunbright PTE-10TSH, etc.).
- the polypeptide is not particularly limited, and examples thereof include collagen, gelatin, albumin, and polylysine.
- examples of the polysaccharide include, but are not limited to, pectin, hyaluronic acid, chitin, chitosan, carboxymethyl chitin, carboxymethyl chitosan, chondroitin sulfate, keratin sulfate, kerato sulfate, heparin, or derivatives thereof.
- suitable combinations of the polysaccharide derivative (active esterified polysaccharide) (A) and the polymer (C) are as follows. In these combinations, the shape (sheet, powder, liquid) can be appropriately selected by referring to the examples described later.
- An ethylene glycol type PEG derivative having a thiol group at two ends, an ethylene glycol type PEG derivative having an amino group at two ends, a trimethylolethane type PEG derivative having a thiol group at three ends, and an amino group at three ends A trimethylolethane type PEG derivative having four pentyl erythritol type PEG derivatives having a thiol group at four terminals, a pentaerythritol type PEG derivative having an amino group at four terminals, a hexaglycerol type PEG derivative having a thiol group at eight terminals, At least one polymer selected from the group consisting of eight glycerol-derived hexaglycerol-type PEG derivatives, albumin, gelatin, collagen, polylysine, pectin, chitosan, chitin and carboxymethyl (CM) chitin (C) a combination of the active esterified pectin.
- An ethylene glycol type PEG derivative having a thiol group at two ends, an ethylene glycol type PEG derivative having an amino group at two ends, a trimethylolethane type PEG derivative having a thiol group at three ends, and an amino group at three ends A trimethylolethane type PEG derivative having four pentyl erythritol type PEG derivatives having a thiol group at four terminals, a pentaerythritol type PEG derivative having an amino group at four terminals, a hexaglycerol type PEG derivative having a thiol group at eight terminals, Active esthetics with at least one polymer (C) selected from the group consisting of hexaglycerol-type PEG derivatives having amino groups at eight ends, albumin, gelatin, collagen, polylysine, pectin, chitosan, chitin and CM chitin The combination of the of CM dextran.
- C Active est
- An ethylene glycol type PEG derivative having a thiol group at two ends, an ethylene glycol type PEG derivative having an amino group at two ends, a trimethylolethane type PEG derivative having a thiol group at three ends, and an amino group at three ends A trimethylolethane type PEG derivative having four pentyl erythritol type PEG derivatives having a thiol group at four terminals, a pentaerythritol type PEG derivative having an amino group at four terminals, a hexaglycerol type PEG derivative having a thiol group at eight terminals, Active esthetics with at least one polymer (C) selected from the group consisting of hexaglycerol-type PEG derivatives having amino groups at eight ends, albumin, gelatin, collagen, polylysine, pectin, chitosan, chitin and CM chitin The combination of the reduction CM pullulan.
- C Active est
- An ethylene glycol type PEG derivative having a thiol group at two ends, an ethylene glycol type PEG derivative having an amino group at two ends, a trimethylolethane type PEG derivative having a thiol group at three ends, and an amino group at three ends A trimethylolethane type PEG derivative having four pentyl erythritol type PEG derivatives having a thiol group at four terminals, a pentaerythritol type PEG derivative having an amino group at four terminals, a hexaglycerol type PEG derivative having a thiol group at eight terminals, Active esthetics with at least one polymer (C) selected from the group consisting of hexaglycerol-type PEG derivatives having amino groups at eight ends, albumin, gelatin, collagen, polylysine, pectin, chitosan, chitin and CM chitin The combination of the reduction CM hydroxyethyl starch.
- SD / AP 20/80 to 98/2 (W / W).
- the compound having a pharmacological action is not particularly limited, but a local anesthetic, antibiotic, antibacterial agent, antifungal agent, antiviral agent, anti-inflammatory agent, vasoconstrictor, steroid hormone, antihistamine, prosta It is preferably at least one selected from the group consisting of glandins and anticancer drugs.
- the local anesthetic is at least one selected from the group consisting of ropivacaine, bupivacaine, levobupivacaine, lidocaine, procaine, tetracaine, cocaine, benzocaine, mepivacaine, prilocaine, dibucaine, chloroprocaine, etidocaine, or a salt thereof. It is preferable.
- dibucaine hydrochloride dibucaine hydrochloride, tetracaine hydrochloride, bupivacaine hydrochloride hydrate, procaine hydrochloride, mepivacaine hydrochloride, lidocaine hydrochloride, levobupivacaine hydrochloride, ropivacaine hydrochloride Salt, etidocaine hydrochloride, prilocaine hydrochloride and chloroprocaine hydrochloride are preferred.
- Antibiotics include ⁇ -lactams such as ampicillin, aminoglycosides such as kanamycin sulfate, tetracyclines such as tetracycline hydrochloride and minocycline, polypeptides such as polymyxin B sulfate, and macrolides such as clarithromycin. And chloramphenicol system such as chloramphenicol.
- antibacterial agents include synthetic antibacterial agents such as a new quinolone series such as tosufloxacin tosylate and a sulfamethoxazole / trimethoprim combination.
- antifungal agents include polyene macrolides such as amphotericin or azoles.
- antiviral agent acyclovir, vitarapin and the like are blended.
- anti-inflammatory drug include diclofenac, celecoxib, dipotassium glycyrrhizinate, lysozyme chloride, and the like.
- vasoconstrictor include naphazoline hydrochloride and dl-methylephedrine hydrochloride.
- steroid hormone agent examples include hydrocortisone butyrate
- examples of the antihistamine agent include diphenhydramine hydrochloride and chlorpheniramine maleate.
- prostaglandin preparations include dinoprost and dinoprostone.
- Anticancer drugs include doxorubicin hydrochloride, pepromycin hydrochloride, nitrogen mustard-N-oxide, cyclofasfamide, thiodepa, carbocon, nimustine hydrochloride, bleomycin hydrochloride, bleomycin sulfate, pepromycin sulfate, aclarubicin hydrochloride, idarubicin hydrochloride, Epirubicin hydrochloride, daunorubicin hydrochloride, pirarubicin hydrochloride, dinostatin stimamarer, neocalcinostatin, etoposide, teniposide, irinotecan hydrochloride, vincristine sulfate, vindesine sulfate, vinblastine sulfate, L-asparaginase, mitoxantrone hydrochloride, cisplatin, carboplatin, nedaplatin , Pen
- the biodegradable gel formed at the time of mixing becomes a suspension form is preferable.
- the administration part of the locally administered agent can be easily confirmed and can be reliably administered to a necessary site.
- the visibility may be expressed when the biodegradable gel becomes cloudy at the time of mixing.
- the biodegradable gel to be formed is in a suspended state, specifically, in a cloudy state so that it can be distinguished from a biological tissue (pink to red).
- a compound having a pharmacological action a compound that becomes cloudy under a predetermined pH condition is used, and it is preferable to prepare a sustained-release topical preparation so that the above-mentioned predetermined pH is obtained upon mixing.
- the biodegradable gel formed upon mixing preferably has a pH of 6.4 to 12, and in particular, the pH is 7.5 to 10.5. Is preferred.
- the biodegradable gel was filled in a cylindrical container having a diameter of 40 mm and a height of 15 mm at a gel temperature of 20 ° C., and a creep meter MODEL RE2-30005B (manufactured by Yamaden Co., Ltd.) was used. It is preferable that the hardness when measuring the hardness when compression measurement is performed at a compression speed of 600 mm / min and a clearance of 5 mm using a resin plunger with a height of 8 mm is 2500 to 20000 N / m 2 . In particular, the hardness of the gel is preferably 6000 to 6500 N / m 2 .
- the sustained-release topical preparation has a release rate of the compound having a pharmacological action after the biodegradable gel is formed for 1 hour and is 8% or more, and has a pharmacological action until 6 hours.
- the compound release rate is preferably 85% or less.
- the sustained release property of the compound having a pharmacological action in the sustained release topical administration agent of the present invention varies depending on the compound having a pharmacological action, but when the compound having a pharmacological action is a local anesthetic, it is mixed. It is preferable that the release rate of the local anesthetic after 1 hour (after administration) is 8% or more, and the release rate of the local anesthetic until 6 hours is 85% or less.
- the release rate (%) is calculated by (the amount of the local anesthetic in the released local administration / the total amount of the local anesthetic in the local administration ⁇ 100).
- the local anesthetic release rate after 72 hours is preferably 80% or more.
- the local anesthetic release rate from administration to 1 hour is preferably 10% or more
- the local anesthetic release rate from 6 hours to 6 hours is preferably less than 70%.
- the local anesthetic release rate up to 72 hours is preferably 82% or more.
- the in vivo degradation rate of the biodegradable gel formed by the sustained-release topical administration agent of the present invention varies depending on the compound having pharmacological action to be used, but one hour has elapsed after mixing (after administration). It is preferable that the decomposition rate until 8 hours is 8% or more, and the decomposition rate until 6 hours elapses is 85% or less.
- the degradation rate of the sustained-release topical preparation is within this range, not only a pharmacological effect is exhibited at an early stage from the administration of the topical preparation to 1 hour, but it also persists after 6 hours.
- the degradation rate of the sustained-release topical administration agent after 72 hours is preferably 80% or more.
- the degradation rate of the sustained-release topical preparation up to 1 hour after administration is 10% or more, and the degradation rate of the sustained-release topical administration agent up to 6 hours is less than 70%. . Furthermore, it is preferable that the degradation rate of the sustained-release topical preparation up to 72 hours is 82% or more.
- the sustained-release topical administration agent of the present invention is a sustained-release topical administration agent used after mixing, as long as it has a compound having a pharmacological action, the above-mentioned crosslinkable polysaccharide derivative, and a pH adjuster.
- the dosage form may be anything. It should be noted that the crosslinkable polysaccharide derivative is more stable in gel formation in the powder than in the liquid.
- Examples of the dosage form of the sustained-release topical administration agent of the present invention include a first agent comprising a powder or liquid containing a compound having a pharmacological action and a second agent comprising a powder or liquid containing a crosslinkable polysaccharide derivative. And a third agent composed of a liquid agent containing a pH adjusting agent is conceivable. And as a 2nd agent, a powder agent is preferable for stability of gel formation. Moreover, it is preferable that the 1st agent containing the compound which has a pharmacological action is a liquid agent from the ease of mixing at the time of use. Moreover, it is preferable that a pH adjuster is a liquid agent from the ease of mixing at the time of use.
- a compound having a pharmacological action and a crosslinkable polysaccharide derivative may be used as powders, and a mixture obtained by mixing the two may be used. Furthermore, it is good also as what added the compound which has a pH adjuster as a liquid agent and has a pharmacological action to this.
- the first agent containing a compound having a pharmacological action is a liquid agent
- the pH adjuster is also a liquid agent
- only the crosslinkable polysaccharide derivative is a powder agent.
- a compound in which a crosslinkable polysaccharide derivative is added to a compound-containing solution having a pharmacological action and then mixed with a pH adjuster that is a solution is suitable.
- the sustained-release topical preparation of the present invention is preferably made into a kit in consideration of convenience during use.
- the sustained release topical administration agent of the present invention contains trehalose.
- Trehalose is preferably present together with the crosslinkable polysaccharide derivative. For this reason, a mixed powder of a crosslinkable polysaccharide derivative and trehalose is preferred.
- Trehalose is a kind of non-reducing disaccharide in which two molecules of D-glucose are linked to each other in 1,1, and glycoside bond is ⁇ , ⁇ -linkage (mycose, ⁇ -D-glucopyranosyl ⁇ -D- glucopyranoside), ⁇ , ⁇ -bonded (neotrehalose, ⁇ -D-glucopyranosyl ⁇ -D-glucopyranoside) and ⁇ , ⁇ -bonded (isotrehalose, ⁇ -D-glucopyranosyl ⁇ -D-glucopyranoside) This means trehalose including the three isomers.
- trehalose isomers may be used alone or in the form of a mixture, but it is preferable to use trehalose whose glycosidic bonds are ⁇ and ⁇ -bonds. Trehalose having glycosidic bonds ⁇ and ⁇ -bonds is most inexpensively available.
- the trehalose of the present invention may be anhydrous trehalose or hydrated trehalose.
- the blending amount of the crosslinkable polysaccharide derivative in the sustained-release topical administration agent of the present invention is 3% by weight (hereinafter,% represents% by weight unless otherwise specified) to 35% with respect to the total amount of the sustained-release topical administration agent. 15 wt% to 25 wt% is particularly preferable.
- the compounding amount of the compound having a pharmacological action in the sustained-release topical administration agent of the present invention varies depending on the compound having a pharmacological action, but is 0.1% by weight (hereinafter referred to as “the sustained-release topical administration agent”). % Represents% by weight unless otherwise specified) to 90%, preferably 3% to 83% by weight.
- the blending amount of the pH adjuster in the sustained-release topical administration agent of the present invention varies depending on the pH adjuster to be used, but is 3% by weight (hereinafter,% is particularly Unless otherwise indicated, it represents 50% by weight), preferably 15% to 40% by weight.
- the amount of trehalose in the sustained-release topical administration agent of the present invention is 10% by weight (hereinafter,% represents% by weight unless otherwise specified) to 50% with respect to the total amount of the sustained-release topical administration agent. It is preferably 15 to 40% by weight.
- the sustained-release topical administration agent of the present invention can further contain widely known additives as long as the properties of the present invention are not impaired.
- additives are not particularly limited, but include curing catalysts, fillers, plasticizers, softeners, stabilizers, dehydrating agents, colorants, anti-sagging agents, thickeners, physical property modifiers, reinforcing agents, thixotropic agents, and deterioration.
- Inhibitors flame retardants, antioxidants, UV absorbers, pigments, solvents, carriers, excipients, preservatives, binders, swelling agents, isotonic agents, solubilizers, preservatives, buffers, diluents, etc. Is mentioned. One or more of these may be included.
- Specific additives include water, physiological saline, pharmaceutically acceptable organic solvents, gelatin, collagen, polyvinyl alcohol, polyvinyl pyrrolidone, carboxyvinyl polymer, sodium carboxymethylcellulose, sodium polyacrylate, sodium alginate, water-soluble Dextran, sodium carboxymethyl starch, pectin, methylcellulose, ethylcellulose, xanthan gum, gum arabic, tragacanth, casein, agar, diglycerin, propylene glycol, polyethylene glycol, petrolatum, paraffin, stearyl alcohol, stearic acid, human serum albumin (HSA) , Mannitol, sorbitol, lactose, PBS, nonionic surfactant, biodegradable polymer, serum-free medium, pharmaceutical supplement
- buffers at physiological pH acceptable in acceptable surfactant or in vivo can be mentioned as objects.
- the polysaccharide derivative used in the sustained-release topical administration agent of the present invention has an active ester group and an active hydrogen-containing group in one molecular chain, and the active ester group and the active hydrogen-containing group react with each other.
- it is a polysaccharide derivative that forms a crosslinked structure by covalent bonding.
- the biodegradable gel that is formed does not use bio-derived materials and has a natural or artificial polysaccharide as the main skeleton, thereby avoiding the risk of infectious diseases and the like.
- the toxicity of the component itself or its degradation product is small, and the biodegradability and absorption are good because the polysaccharide is the main skeleton.
- B Group amount of carboxy group and carboxymethyl group (mmol / g)
- the reaction solvent is DMSO
- the electrophilic group introducing agent is N-hydroxysuccinimide (NHS) (manufactured by Wako Pure Chemical Industries, Ltd.)
- NHS N-hydroxysuccinimide
- EDC 1-ethyl-3-dimethylaminopropylcarbodiimide hydrochloride
- 0.1, 0.2, 0.5, 1.0, and 2.5 mM NHS standard aqueous solutions were prepared.
- 0.2 mL of 2N sodium hydroxide aqueous solution was added to 1 mL of each NHS standard aqueous solution, and it heated at 60 degreeC and stirred for 10 minutes.
- 1.5 mL of 0.85N hydrochloric acid and 0.5 mL of 0.5% FeCl 3 / 1N hydrochloric acid solution were added, and the absorbance at an absorption wavelength of 500 nm was measured using a spectrophotometer (FeCl 3 , Wako Pure). Yakuhin Kogyo).
- Example 2 Preparation of sustained-release topical agent
- the freeze-dried product of NHS-modified carboxymethyldextrin of Synthesis Example 1 was used.
- a freeze-dried product of trehalose was used.
- Dry sodium carbonate and dry sodium hydrogen carbonate were used as pH adjusters.
- ropivacaine hydrochloride monohydrate (molecular weight 328.88) was used.
- a mixture of NHS-modified carboxymethyldextrin lyophilized product and trehalose at a weight ratio of 1: 1 was prepared, and then 2.5 g of a product sterilized by electron beam (powder A: first agent) was prepared.
- a product sterilized by electron beam prowder A: first agent
- ropivacaine hydrochloride monohydrate 3.6 ml of 59 mg / ml ropivacaine hydrochloric acid aqueous solution
- a pH adjuster liquid B: third agent
- the sustained release local administration agent (Example) of the present invention consisting of the first agent, the second agent (liquid A) and the third agent (liquid B) was prepared.
- Example 1 Using the sustained-release topical preparation of Example, a drug sustained-release confirmation test was conducted.
- the first agent (NHS-modified carboxymethyldextrin freeze-dried product and trehalose mixture, 2.5 g) was added to the second agent (liquid A: 3.6 mg of 59 mg / mL ropivacaine hydrochloric acid aqueous solution), and dissolved to give the first agent.
- a liquid (first agent and second agent mixture) was prepared.
- Example 2 56 ⁇ L of water was added to 112 ⁇ L of the third agent (liquid B, pH adjusting agent) to prepare a prepared third agent (prepared liquid B, pH adjusting agent).
- a biodegradable gel (gel 2) containing a drug was prepared by adding 800 ⁇ L of the same first solution as in Formulation Example 1 to 168 ⁇ L of this prepared third agent and stirring with a vortex mixer until gelled. The time required for gelation was 1 minute 8 seconds. The pH of the gel was 6.73, and the gel was cloudy and visible.
- Example 3 84 ⁇ L of water was added to 84 ⁇ L of the third agent (liquid B, pH adjusting agent) to prepare a prepared third agent (prepared liquid B, pH adjusting agent).
- a biodegradable gel (gel 3) containing a drug was prepared by adding 800 ⁇ L of the same first solution as in Formulation Example 1 to 168 ⁇ L of this prepared third agent and stirring with a vortex mixer until gelled. The time required for gelation was 2 minutes and 0 seconds. The pH of the gel was 6.42, and the gel was cloudy and visible.
- the sodium lactate-containing infusion preparation contains 3.0 g of sodium chloride, 0.150 g of potassium chloride, 0.10 g of calcium chloride hydrate, and 3.10 g of L-sodium lactate solution in 500 ml of active ingredients. -1.55 g) as sodium lactate.
- first liquid first agent and second agent mixture
- third agent liquid B, pH adjusting agent
- a biodegradable gel (gel 2) containing a drug was prepared by adding 205 ⁇ L of the same first solution as in Formulation Example 1 to 43 ⁇ L of this prepared third agent and stirring with a vortex mixer until gelled.
- 22 ⁇ L of water is added to 22 ⁇ L of the third agent (liquid B, pH adjusting agent) to prepare the third agent (prepared liquid B, pH adjusting agent) so that the mixing ratio is the same as that of formulation example 3.
- a biodegradable gel (gel 3) containing a drug was prepared by adding 205 ⁇ L of the same first solution as in Formulation Example 1 to 44 ⁇ L of this prepared third agent and stirring with a vortex mixer until gelled.
- the biodegradable gel (gel 1), biodegradable gel (gel 2), and biodegradable gel (gel 3) used in the sustained release confirmation test 1 were mixed with 968 ⁇ L of PBS (phosphate buffer). Solution, pH 7.4) Add 9mL, soak in a 37 ° C water bath, take 0.5mL after 10 minutes, 30 minutes, 1 hour, 2 hours, 3 hours, 6 hours, 24 hours, 1 week, respectively. The mobile phase was added to 5 mL to make a sample solution. The release rate was calculated by measuring the peak area of ropivacaine according to the US Pharmacopoeia “Ropivacaine Hydrochloride Injection: Assay” for 20 ⁇ L of each sample solution. The results were as shown in Table 2.
- the sustained-release topical administration agent of the present invention is a gel. It was confirmed that the drug was released by the decomposition of. In the sustained-release topical preparation of the present invention, since the release of the drug and the degradation / disappearance of the gel occur simultaneously, only the gel does not remain after all the drug is released.
- Example 2 Using rats, the presence of drug-containing biodegradable gel was confirmed and the plasma drug concentration was measured. Under isoflurane anesthesia, the abdomen of male SD rat (body weight: about 300 g) was incised and peeled to expose the subcutaneous tissue (abdominal wall), and the biodegradable gel of Formulation Example 1 (containing 8.75 mg as ropivacaine hydrochloride, pH 7) .56) was administered. The administered biodegradable gel was cloudy, and the administration site and the non-administration site could be easily confirmed visually.
- the change in plasma ropivacaine concentration was as shown in FIG. The 3-hour value after administration became the highest and gradually decreased, and was about 200 ng / mL at 12 hours after administration, and was below the lower limit of quantification at 18 hours.
- Example 3 Using rabbits, confirmation of residual drug-containing biodegradable gel and plasma drug concentration were measured. Under isoflurane anesthesia, the female Kbl: JW rabbit (weight approximately 3 kg) was incised and detached from the back to create a subcutaneous pocket, and the biodegradable gel (containing 70 mg of ropivacaine hydrochloride) of the above Formulation Example 1 was administered ( 6 animals in total). The administered biodegradable gel was cloudy, and the administration site and the non-administration site could be easily confirmed visually.
- Plasma ropivacaine concentration rises rapidly after administration, reaches the highest level for 3-6 hours after administration, remains slightly flat thereafter, and is below the lower limit of quantitation (10 ng / ml) after 48 hours after administration. It was.
- the sustained-release topical preparation of the present invention is as follows.
- a sustained-release topical agent used after mixing The sustained-release topical preparation has a compound having a pharmacological action and at least one active ester group introduced into a polysaccharide side chain and capable of reacting with an active hydrogen-containing group, and the active ester group and the active hydrogen
- the sustained-release topical agent forms the biodegradable gel at the time of mixing, and the pH of the biodegradable gel formed is 5.
- a sustained-release topical agent that is 5 or more.
- the sustained-release topical administration agent of the present invention forms a biodegradable gel containing a compound having a pharmacological action at the administered site after mixing. Since it is gelled, it stays at the administration site, and the compound having a pharmacological action is gradually released as the gel disintegrates over time. For this reason, the compound which has a pharmacological action can be administered in the administration site for a long time until the gel collapses. In addition, since the release of the compound is caused by the decomposition of the gel, the disappearance of the compound and the gel ends substantially at the same time, and the gel does not remain. Moreover, the formed gel has good adhesion to the surface of the living body and is excellent in flexibility.
- the above embodiment may be as follows.
- the compound having the pharmacological action is a local anesthetic, antibiotic, antibacterial agent, antifungal agent, antiviral agent, anti-inflammatory agent, vasoconstrictor, steroid hormone, antihistamine, prostaglandin, anticancer
- the local anesthetic is at least one selected from the group consisting of ropivacaine, bupivacaine, levobupivacaine, lidocaine, procaine, tetracaine, cocaine, benzocaine, mepivacaine, prilocaine, dibucaine, chloroprocaine, etidocaine, or a salt thereof.
- the pH adjuster is an inorganic base or an organic base and is any one of sodium hydroxide, sodium carbonate, sodium bicarbonate, a phosphate buffer, and an acetic acid-ammonia buffer.
- the sustained-release topical administration agent according to any one of (5).
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Abstract
Description
混合後使用される徐放性局所投与剤であって、前記徐放性局所投与剤は、薬理作用を有する化合物と、多糖側鎖に導入された、活性水素含有基と反応しうる活性エステル基を少なくとも1つ有し、前記活性エステル基と前記活性水素含有基との共有結合による架橋物を形成しうる架橋性多糖誘導体と、前記架橋性多糖誘導体と非混合状態であり、かつ混合時において、前記化合物を含有する生体内分解性ゲルを形成するpH条件とするためのpH調整剤とを有し、前記徐放性局所投与剤は、前記混合時において、前記生体内分解性ゲルを形成し、かつ形成される前記生体内分解性ゲルのpHが5以上である徐放性局所投与剤。
本発明の徐放性局所投与剤は、混合後使用される徐放性局所投与剤である。そして、徐放性局所投与剤は、薬理作用を有する化合物と、多糖側鎖に導入された、活性水素含有基と反応しうる活性エステル基を少なくとも1つ有し、活性エステル基と活性水素含有基との共有結合による架橋物を形成しうる架橋性多糖誘導体と、架橋性多糖誘導体と非混合状態であり、かつ混合時において、薬理作用を有する化合物を含有する生体内分解性ゲルを形成するpH条件とするためのpH調整剤とを有する。徐放性局所投与剤は、混合時において、薬理作用を有する化合物を含有する生体内分解性ゲルを形成し、さらに、形成される生体内分解性ゲルのpHが5以上である。
なお「1分子鎖」または「分子内」の分子とは、共有結合により連続した結合で繋がった範囲の1つの分子を意味する。
本発明において、多糖誘導体の活性エステル基は、1種単独でも2種以上が存在していてもよい。
このような活性エステル基の中でも、スクシンイミドエステル基が好ましい。
上記デキストランは、代用血漿剤として使用されている。デキストランとしては、アマシャムバイオサイエンス社(日本)の「Dextran T fractions」、プルランは林原社(日本)の「Pullulan PI-20」が挙げられる。プルランは、経口薬を含む医薬添加剤として使用されており、エンドトキシン等の生物学的コンタミネーションが少ないものが好適である。
いずれの多糖も、本発明においては、一般的に商業流通しているものを利用できる。上記医療用途で実績のある多糖は、本発明においては安全性面で好適に利用できる多糖である。
上記のような酸基含有多糖のカルボキシ基および/またはカルボキシメチル基を活性エステル化するに際して、酸基含有多糖は、単独で使用しても良いし、2種以上のものを併用して使用しても良い。
したがって、この方法で活性エステル化される酸基含有多糖として、上記に例示した多糖のうちでも、60℃~120℃の間の温度で非プロトン性極性溶媒に溶解するものが好ましく使用される。具体的に、求電子性基導入のための反応に用いられる多糖は、非プロトン性極性溶媒への溶解性の点から、カルボキシ基またはカルボキシメチル基が酸型であることが好ましい。「酸型」とは、カルボキシ基またはカルボキシメチル基のカウンターカチオン種がプロトンであることをいう。酸型のカルボキシ基を有する多糖を酸型 (原料) 多糖という。例えば、カルボキシ基を有する多糖であるペクチンを酸型ペクチンという。酸型のカルボキシメチル基を有するカルボキシメチルデキストランを酸型カルボキシメチル(CM)デキストラン(酸型CMデキストラン)という。「酸型」は、カウンターカチオン種がプロトンであり、塩形態ではない点で前記「非塩型」と同義である。
乾燥工程においては、前記精製工程で得られた多糖誘導体から洗浄溶媒を除去するため、通常使用される方法により乾燥させればよい。
活性エステル基導入率は、例えば、Biochemistry Vol. 14, No.7(1975), p1535-1541に記載の方法により決定することができる。
特に、上記100%未満の活性エステル基の導入率で活性エステル基が導入された場合に残存する原料多糖の有するカルボキシ基および/またはカルボキシメチル基を有していてもよい。
この系では、用時両者を混合して、最終的な多糖誘導体の濃度が0.1~60%(W/V)の混合水溶液とすることができる。また、多糖誘導体の濃度が1~80%(W/V)の水溶液に、用時、pH調整剤(B)の塩を添加して溶解させながら混合して、最終的な多糖誘導体の濃度が0.1~80%(W/V)の混合水溶液からなるものも挙げることができる。混合は、通常の混合方法を選択することができるが、混合状態が均一になるまで行うことが好ましく、所望の反応が進行する程度での均一さであればよい。
局所麻酔薬としては、ロピバカイン、ブピバカイン、レボブピバカイン、リドカイン、プロカイン、テトラカイン、コカイン、ベンゾカイン、メピバカイン、プリロカイン、ジブカイン、クロロプロカイン、エチドカインまたはこれらの塩からなる群より選択された少なくとも1種であることが好ましい。特に、形成される生分解性ゲルの視認性の点から、ジブカイン塩酸塩、テトラカイン塩酸塩、ブピバカイン塩酸塩水和物、プロカイン塩酸塩、メピバカイン塩酸塩、リドカイン塩酸塩、レボブピバカイン塩酸塩、ロピバカイン塩酸塩、エチドカイン塩酸塩、プリロカイン塩酸塩、クロロプロカイン塩酸塩が好ましい。
また、使用する薬理作用を有する化合物として、形成される生体内分解性ゲルが、懸濁状態、具体的には、生体組織(ピンク~赤)と区別し得るよう白濁状態となるものが好ましい。薬理作用を有する化合物としては、所定のpH条件にて白濁するものを用い、かつ、徐放性局所投与剤として、混合時に上記の所定pHとなるように調製することが好ましい。
本発明の徐放性局所投与剤における薬理作用を有する化合物の徐放性は、含有する薬理作用を有する化合物によっても異なるが、含有する薬理作用を有する化合物が局所麻酔剤の場合には、混合後(投与後)1時間経過時までの局所麻酔薬の放出率が、8%以上であり、かつ、6時間経過時までの局所麻酔薬の放出率が、85%以下であることが好ましい。放出率の数値がこの範囲にあることにより、ゲル状局所麻酔薬投与から1時間までの早期の段階で鎮痛効果が発揮されるだけでなく、6時間後も鎮痛効果が持続する。尚、放出率(%)は、(放出した局所投与剤中の局所麻酔薬量/局所投与剤中の局所麻酔薬総量×100)で計算される。さらには、72時間後の局所麻酔薬の放出率が、80%以上であることが好ましい。特に、投与から1時間までの局所麻酔薬の放出率が、10%以上であり、かつ、6時間までの局所麻酔薬の放出率が、70%未満であることが好ましい。さらには、72時間までの局所麻酔薬の放出率が、82%以上であることが好ましい。
トレハロースは、2分子のD-グルコースが1,1結合した形の非還元性二糖の一種であり、グリコシド結合がα、α-結合であるもの(ミコース、α-D-glucopyranosyl α-D-glucopyranoside)、α、β-結合であるもの(ネオトレハロース、β-D-glucopyranosyl α-D-glucopyranoside)及びβ、β-結合であるもの(イソトレハロース、β-D-glucopyranosyl β-D-glucopyranoside)の3種類の異性体を含めたトレハロースを意味するものである。
本発明の徐放性局所投与剤における薬理作用を有する化合物の配合量は、薬理作用を有する化合物によっても相違するが、徐放性局所投与剤の全量に対して、0.1重量%(以下%は特に断りのない限り重量%を表わす)~90%が好ましく、3重量%~83重量%が特に好ましい。
また、本発明の徐放性局所投与剤におけるpH調整剤の配合量は、使用するpH調整剤によっても相違するが、徐放性局所投与剤の全量に対して3重量%(以下%は特に断りのない限り重量%を表わす)~50%がよく、15重量%~40重量%が好ましい。
また、本発明の徐放性局所投与剤におけるトレハロースの配合量は、徐放性局所投与剤の全量に対して10重量%(以下%は特に断りのない限り重量%を表わす)~50%がよく、15重量%~40重量%が好ましい。
(1) 原料多糖(酸型多糖)の調製
活性エステル化多糖誘導体の原料となる原料多糖としてカルボキシメチルデキストリン(酸型CMデキストリン)を調製した。
デキストリン(和光純薬工業社製、重量平均分子量25000)10gを、精製水62.5gに溶解させた後、36%水酸化ナトリウム水溶液(W/V)(水酸化ナトリウム、和光純薬工業社製)62.5gを添加し、25℃で90分間攪拌し溶解した。
上記(1)で得られた酸型CMデキストラン(原料多糖)について、これらのカルボキシ基、あるいはカルボキシメチル基の定量を行った。原料多糖0.2g(A(g))を秤取り、0.1mol/L水酸化ナトリウム水溶液20mLと80vol%メタノール水溶液10mLとの混合溶液に添加し、25℃で3時間攪拌した。得られた溶液に、指示薬として1.0%フェノールフタレイン(W/V)/90vol%エタノール水溶液を3滴添加し、0.05mol/L硫酸を使用して酸塩基逆滴定を行い、0.05mol/L硫酸の使用量(V1 mL)を測定した(フェノールフタレイン、和光純薬工業社製)。また、原料多糖を添加しない以外は同様にして行ったブランクでの0.05mol/L硫酸の使用量(V0 mL)を測定した。下記式(1)に従い、原料多糖のカルボキシ基およびカルボキシメチル基の基量(Bmmol/g)を算出したところ、0.60mmol/gであった。なお、使用した0.1mol/L水酸化ナトリウム水溶液、0.05mol/L硫酸の力価は、ともに1.00であった。
B=(V0-V1)×0.1÷A ・・・・・・(1)
A:原料多糖の質量(g)
B:カルボキシ基およびカルボキシメチル基の基量(mmol/g)
酸型CMデキストランの活性エステル化反応には、反応溶媒はDMSO、求電子性基導入剤はN-ヒドロキシスクシンイミド(NHS)(和光純薬工業社製)、脱水縮合剤は1-エチル-3-ジメチルアミノプロピルカルボジイミド塩酸塩(EDC)(和光純薬工業社製)を使用し、活性エステル化多糖(多糖誘導体)を調製した。
上記(1)で得られた酸型CMデキストラン(カルボキシメチル基量0.60mmol/g)2.0gを、DMSO200gに溶解した。その後、NHS2.8g(24.3mmol)とEDC4.6g(24.0mmol)を添加して、25℃で19時間攪拌した。反応溶液をアセトン、メタノール混合溶媒4Lに滴下し、吸引ロートを用いて析出物を回収した。アセトン、メタノール混合溶媒4Lを使用して得られた析出物を洗浄して、減圧乾燥した。これにより、活性エステル化CMデキストランを調製した。
(3)で得られた活性エステル化CMデキストランについて、以下のようにして求めたNHS導入量は、0.70mmol/gであった。
NHS導入量は、多糖誘導体の単位重量あたりに存在するNHS含有量である。
Y=αX+β ・・・・・・(2)
X:NHS濃度(mM)
Y:波長500nmにおける吸光度
α=0.178 (傾き)
β=0.021 (切片)
r=0.995 (相関係数)
吸光度を元にNHS濃度、X(mM)が算出される。
NHS導入量(mmol/g)=(D×H)×0.001/C・・・・(3)
上記で得られた活性エステル化多糖が自己架橋性であることを、以下の試験により確認した。容量10mLの清浄試験管(ラルボLT-15100、テルモ社製)に、活性エステル化多糖0.2gを秤取り、純水1mLを添加して混合した。次に、pH調整剤として8.3%炭酸水素ナトリウム水溶液(W/V)(炭酸水素ナトリウム、和光純薬工業社製)1mL(pH8.3)を添加し、試験管ミキサー(MT-31、ヤマト科学社製)を用いて約2,000rpmで約1分間混合した。その混合前後での試験管内容物の状態を目視にて確認した。これにより、活性エステル化CMデキストランは、混合後の試験管内容物が塊状物(含水ゲル)になっており、「自己架橋性あり」と判定した。
(徐放性局所投与剤の作成)
架橋性多糖誘導体として、合成例1のNHS修飾カルボキシメチルデキストリンの凍結乾燥物を用いた。トレハロースの凍結乾燥物を用いた。pH調整剤として、乾燥炭酸ナトリウム、乾燥炭酸水素ナトリウムを用いた。薬物として、ロピバカイン塩酸塩一水和物(分子量328.88)を用いた。
実施例の徐放性局所投与剤を用いて、薬物の徐放性確認試験を行った。
(配合例1)
第2剤(液体A:59mg/mLロピバカイン塩酸水溶液3.6ml)に、第1剤(NHS修飾カルボキシメチルデキストリン凍結乾燥物とトレハロース混合物、2.5g)を添加し、溶解することにより、第1液(第1剤と第2剤混合物)を調製した。15mlファルコンチューブに、第1液800μLを取り,上記の第3剤(液体B、pH調整剤)168μL加え、ゲル化するまでボルテックスミキサーで攪拌することにより、薬物を含有する生体内分解性ゲル(ゲル1)を作成した。ゲル化までの所要時間は、15秒であった。ゲルのpHは、7.56であり、ゲルは白濁しており視認可能であった。日本電色工業株式会社SpectrophotometerSA4000で反射光を測定。L=76.60、a=-1.90、b=27.52。
上記の第3剤(液体B、pH調整剤)112μLに,水56μLを加えて調製第3剤(調製液体B、pH調整剤)を作成した。この調製第3剤168μLに、配合例1と同じ第1液800μLを加え,ゲル化するまでボルテックスミキサーで攪拌することにより、薬物を含有する生体内分解性ゲル(ゲル2)を作成した。ゲル化までの所要時間は、1分8秒であった。ゲルのpHは、6.73あり、ゲルは白濁しており視認可能であった。
上記の第3剤(液体B、pH調整剤)84μLに,水84μLを加えて調製第3剤(調製液体B、pH調整剤)を作成した。この調製第3剤168μLに、配合例1と同じ第1液800μLを加え,ゲル化するまでボルテックスミキサーで攪拌することにより、薬物を含有する生体内分解性ゲル(ゲル3)を作成した。ゲル化までの所要時間は、2分0秒であった。ゲルのpHは、6.42であり、ゲルは白濁しており視認可能であった。
20300U/Lα-アミラーゼ(from human saliva)2.5mLを正確に量り,乳酸ナトリウム含有輸液剤を加えて正確に50mlとし、アミラーゼ溶液を作成した。なお、乳酸ナトリウム含有輸液剤は、有効成分 500ml中に、塩化ナトリウムを3.0g、塩化カリウムを0.150g、塩化カルシウム水和物を0.10g、L-乳酸ナトリウム液を3.10g(L-乳酸ナトリウムとして1.55g)を含有している。
配合例1と同じ配合比率となるように、第1液(第1剤と第2剤混合物)205μLを直径2.1cmのスクリュー管瓶に取り、第3剤(液体B、pH調整剤)43μLを加え、ゲル化するまでボルテックスミキサーで攪拌し、薬物を含有する生体内分解性ゲル(ゲル1)を作成した。
同様に、配合例3と同じ配合比率となるように、上記の第3剤(液体B、pH調整剤)22μLに,水22μLを加えて調製第3剤(調製液体B、pH調整剤)を作成した。この調製第3剤44μLに、配合例1と同じ第1液205μLを加え,ゲル化するまでボルテックスミキサーで攪拌することにより、薬物を含有する生体内分解性ゲル(ゲル3)を作成した。
徐放性確認試験1にて用いた生体内分解性ゲル(ゲル1)、生体内分解性ゲル(ゲル2)および生体内分解性ゲル(ゲル3)のそれぞれのゲル968μLにPBS(リン酸緩衝液、pH7.4)9mLを加え,37℃の水浴に浸漬し,10分,30分,1時間,2時間,3時間,6時間,24時間,1週間後に0.5mLずつ採取し,それぞれに移動相を加えて5mLとし,試料溶液とした.この試料溶液それぞれ20μLにつき,米国薬局方「Ropivacaine Hydrochloride Injection:Assay」に従ってロピバカインのピーク面積を測定することにより、放出率を算出した。結果は、表2に示す通りであった。
ラットを用いて、薬物含有生体内分解性ゲルの残存確認と血漿中薬物濃度を測定した。
イソフルラン麻酔下で雄SDラット(体重約300g)の腹部を切開・剥離し皮下組織(腹壁)を露出させ、上述の配合例1の生体内分解性ゲル(ロピバカイン塩酸塩として8.75mg含有、pH7.56)を投与した。投与した生体内分解性ゲルは、白濁しており、投与部位と非投与部位を視認により容易に確認できた。
血漿中ロピバカイン濃度変化は、図1に示す通りであった。投与後3時間値が最も高くなり、緩やかに低下し、投与後12時間に約200ng/mL、18時間では定量下限値未満になった。
ウサギを用いて、薬物含有生体内分解性ゲルの残存確認と血漿中薬物濃度を測定した。
イソフルラン麻酔下で雌Kbl:JWウサギ(体重約3kg)の背部を切開・剥離し皮下ポケットを作製し、上述の配合例1の生体内分解性ゲル(ロピバカイン塩酸塩として70mg含有)を投与した(計6匹)。投与した生体内分解性ゲルは、白濁しており、投与部位と非投与部位を視認により容易に確認できた。
血漿中ロピバカイン濃度変化は、図2に示す通りであった。血漿中ロピバカイン濃度は、投与後速やかに上昇し、投与後3-6時間値が最も高くなり、その後はやや横ばいを維持し、投与後48時間以降では定量下限値(10ng/ml)未満になった。
(1) 混合後使用される徐放性局所投与剤であって、
前記徐放性局所投与剤は、薬理作用を有する化合物と、多糖側鎖に導入された、活性水素含有基と反応しうる活性エステル基を少なくとも1つ有し、前記活性エステル基と前記活性水素含有基との共有結合による架橋物を形成しうる架橋性多糖誘導体と、前記架橋性多糖誘導体と非混合状態であり、かつ混合時において、前記化合物を含有する生体内分解性ゲルを形成するpH条件とするためのpH調整剤とを有し、前記徐放性局所投与剤は、前記混合時において、前記生体内分解性ゲルを形成し、かつ形成される前記生体内分解性ゲルのpHが5以上である徐放性局所投与剤。
(2) 前記徐放性局所投与剤は、前記混合時に形成される前記生体内分解性ゲルのpHが、6.4~12である上記(1)に記載の徐放性局所投与剤。
(3) 前記徐放性局所投与剤は、前記生体内分解性ゲル形成後から1時間経過時までの前記化合物の放出率が、10%以上であり、6時間経過時までの前記化合物の放出率が、70%未満である上記(1)または(2)に記載の徐放性局所投与剤。
(4) 前記薬理作用を有する化合物は、局所麻酔薬、抗生物質、抗菌薬、抗真菌薬、抗ウイルス薬、抗炎症薬、血管収縮薬、ステロイドホルモン、抗ヒスタミン薬、プロスタグランジン、抗癌薬からなる群から選択された少なくとも1種である上記(1)ないし(3)のいずれかに記載の徐放性局所投与剤。
(5) 前記局所麻酔薬は、ロピバカイン、ブピバカイン、レボブピバカイン、リドカイン、プロカイン、テトラカイン、コカイン、ベンゾカイン、メピバカイン、プリロカイン、ジブカイン、クロロプロカイン、エチドカインまたはこれらの塩からなる群より選択された少なくとも1種である上記(4)に記載の徐放性局所投与剤。
(6) 前記pH調整剤は、無機塩基または有機塩基であり、水酸化ナトリウム、炭酸ナトリウム、炭酸水素ナトリウム、リン酸系緩衝剤、酢酸-アンモニア系緩衝剤のいずれかである上記(1)ないし(5)のいずれかに記載の徐放性局所投与剤。
(7) 前記徐放性局所投与剤は、前記生体内分解性ゲルは、懸濁状となることによる視認性を有する上記(1)ないし(6)のいずれかに記載の徐放性局所投与剤。
(8) 前記生体内分解性ゲルは、混合時において白濁するものである上記(7)に記載の徐放性局所投与剤。
Claims (8)
- 混合後使用される徐放性局所投与剤であって、
前記徐放性局所投与剤は、薬理作用を有する化合物と、多糖側鎖に導入された、活性水素含有基と反応しうる活性エステル基を少なくとも1つ有し、前記活性エステル基と前記活性水素含有基との共有結合による架橋物を形成しうる架橋性多糖誘導体と、前記架橋性多糖誘導体と非混合状態であり、かつ混合時において、前記化合物を含有する生体内分解性ゲルを形成するpH条件とするためのpH調整剤とを有し、前記徐放性局所投与剤は、前記混合時において、前記生体内分解性ゲルを形成し、かつ形成される前記生体内分解性ゲルのpHが5以上であることを特徴とする徐放性局所投与剤。 - 前記徐放性局所投与剤は、前記混合時に形成される前記生体内分解性ゲルのpHが、6.4~12である請求項1に記載の徐放性局所投与剤。
- 前記徐放性局所投与剤は、前記生体内分解性ゲル形成後から1時間経過時までの前記化合物の放出率が、10%以上であり、6時間経過時までの前記化合物の放出率が、70%未満である請求項1または2に記載の徐放性局所投与剤。
- 前記薬理作用を有する化合物は、局所麻酔薬、抗生物質、抗菌薬、抗真菌薬、抗ウイルス薬、抗炎症薬、血管収縮薬、ステロイドホルモン、抗ヒスタミン薬、プロスタグランジン、抗癌薬からなる群から選択された少なくとも1種である請求項1ないし3のいずれかに記載の徐放性局所投与剤。
- 前記局所麻酔薬は、ロピバカイン、ブピバカイン、レボブピバカイン、リドカイン、プロカイン、テトラカイン、コカイン、ベンゾカイン、メピバカイン、プリロカイン、ジブカイン、クロロプロカイン、エチドカインまたはこれらの塩からなる群より選択された少なくとも1種である請求項4に記載の徐放性局所投与剤。
- 前記pH調整剤は、無機塩基または有機塩基であり、水酸化ナトリウム、炭酸ナトリウム、炭酸水素ナトリウム、リン酸系緩衝剤、酢酸-アンモニア系緩衝剤のいずれかである請求項1ないし5のいずれかに記載の徐放性局所投与剤。
- 前記徐放性局所投与剤は、前記生体内分解性ゲルは、懸濁状となることによる視認性を有する請求項1ないし6のいずれかに記載の徐放性局所投与剤。
- 前記生体内分解性ゲルは、混合時において白濁するものである請求項7に記載の徐放性局所投与剤。
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AU2016352007A AU2016352007B2 (en) | 2015-11-12 | 2016-11-01 | Sustained-release topically administered agent |
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US20200054754A1 (en) * | 2017-05-11 | 2020-02-20 | Steadymed Ltd. | Enhanced solubility drug-containing formulations |
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