JP2005281231A - Small intestine target-type oral dds preparation - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a drug preparation effective for increasing the absorption of a peptide-protein drug which is hydrolyzed with digestive enzymes in the digestive tracts after oral administration, having low membrane permeability owing to its polymer nature and unable to attain a sufficient absorption quantity. <P>SOLUTION: A solidified preparation system for promoting the absorption of a drug contains (a) an absorbefacient, (b) fine particles, (c) a peptide-protein drug, and as necessary, (d) a stabilizer. The system is a solid preparation effective for delivering the peptide-protein drug to the absorption cells of small intestine while suppressing the hydrolysis with digestive enzymes by holding the absorbefacient and the peptide-protein drug with the fine particles, and achieving efficient absorption of the peptide-protein drug by forming a high concentration gradient. In the case of more strongly suppressing the hydrolysis of the peptide-protein drug with the digestive enzymes, it is effective to add a food protein or a hydrolase inhibiting agent, or the like, as the stabilizer. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to a microparticulate DDS preparation comprising an absorption promoter such as a surfactant and a peptide / protein drug, which protects the peptide / protein drug from digestive enzymes and promotes drug absorption. More specifically, the present invention absorbs drugs that are difficult to absorb from the digestive tract after oral administration due to significant hydrolysis by digestive enzymes, etc., such as peptide and protein drugs, and low permeability through the absorbent membrane. In order to improve the absorption rate of peptides and protein drugs after oral administration, protect the drug from hydrolase in the gastrointestinal tract and form a locally high drug concentration gradient between the absorbing cells and the system. The present invention relates to a microparticulate DDS preparation for the purpose of promoting drug absorption including an absorption enhancer, which is used for enhancing (bioavailability) and drug efficacy.

  In addition to Emisphere's capric acid derivatives and emulsion technology, there are numerous oral DDS technologies, including various patented technologies described and cited in the following technical papers. (Non-patent documents 1 to 6). However, the product has not been commercialized due to insufficient absorption. Patent Documents 1 to 15 describe solidified preparations containing a surfactant, a porous carrier and a drug, but no satisfactory effect has been obtained for oral absorption of peptide / protein drugs.

  Peptides and protein drugs such as erythropoietin, granulocyte growth factor G-CSF, insulin, calcitonin, interferon, various interleukins, vasopressin and its derivatives have been abandoned due to low absorption after oral administration. Large doses must be used even if they are administered orally. In addition, there are many peptides and protein drugs that are currently under development, which have high bioactivity as well as these peptide and protein drugs, but they are commercialized because of their low absorption after oral administration. It has been buried without being buried. In order to increase the bioavailability of peptides and protein drugs that cannot be expected to be sufficiently effective by oral administration due to the low bioavailability of conventional administration methods, Takada first adheres to the gastrointestinal mucosa and starts with peptide and protein drugs. Invented the DDS technology (registered trademark GI-MAPS) that enhances the oral absorbability of difficult and low-absorbable drugs (PCT / JP99 / 06602, An Oral Formulation for Gastrointestinal Drug Delivery, International Publication No. WO00 / 32172). Basically, this technique is an asymmetric (asymmetrical, non-spherical) patch. The feature of GI-MAPS is that after oral administration, it adheres to the absorption membrane surface of the gastrointestinal tract to form a closed space, thereby deriving a high drug concentration gradient between the system and the absorbing cells, resulting in high drug absorption. It is a system to get sex. Most drugs are absorbed by simple diffusion (passive transport). When a drug molecule is absorbed by simple diffusion, a high concentration gradient of the drug molecule between the luminal side of the digestive tract and the small intestine absorbing cells is a driving force for absorption. Therefore, if a high concentration gradient of the drug is formed between the digestive tract-absorbing cells and the inside of the system, the absorption membrane permeability of the drug increases. This basic design concept of GI-MAPS is supported by recent research results.

Hiroaki Okada, Nikkei Biobusiness, November 2003, pages 148-151 Isabel Gomez-Orellana and Duncan R. Paton, Advances in the oral delivery of proteins.Exptl. Opin. Ther. Patents, 9, 247-253, 1999. Rakhi B. Shah, Fakhrul Ahsan & Mansoor A. Khan, Oral delivery of proteins: Progress and prognostication. Crit. Rev. Ther. Drug Carrier Systems, 19, 135-169, 2002. J. Gordon Still, Development of oral insulin: Progress and current status.Diabetes Metab. Res. Rev., 18, suppl. 1, S29-S37, 2002. Y. H. Lee, B. A. Perry, J. P. Sutyak, W. Stern and P. J. Sinko, Regional differences in intestinal spreading and pH recovery and the impact on salmon calcitonin in dogs. Pharm. Res., 17, 284-290, 2000. PJ Sinko, YH Lee, V. Makhey, GD Leesman, JP Sutyak, H. Yu, B. Perry, CL Smith, P. Hu, EJ Wagner, LM Falzone, LT McWhorter, JP Gilligan and W. Stern, Biopharmaceutical approaches for developing and assessing oral peptide delivery strategies and systems: in vitro permeability and in vivo oral absorption of salmon calcitonin (sCT). Pharm. Res., 16, 527-533, 1999. JP-A-6-16537 Japanese Patent Laid-Open No. 10-17495 EP448091 A1 publication International Publication No. WO01 / 10410 International Publication WO01 / 95912 JP 2000-16934 A International Publication WO01 / 41737 International Publication WO01 / 180 JP 2001-316248 A International Publication WO00 / 27393 JP-A-4-159222 JP 58-213073 A International Publication No. WO99 / 53965 GB2114885A Japanese Patent Laid-Open No. 9-216815 International Publication WO00 / 32172

  GI-MAPS is composed of three layers: a basal layer, a drug retaining layer, and a surface layer. Among these, the basal layer is formed of a water-insoluble polymer film, and completely blocks the attack of digestive enzymes such as proteolytic enzymes from the luminal side of the digestive tract. On the other hand, the surface layer is formed by a pH-sensitive polymer film that dissolves at the pH of the duodenum, jejunum, and ileum, etc., and is most advantageous for absorption of peptide and protein drugs produced by methods such as genetic recombination. A secure target site and a lined adhesive layer facilitates adhesion of the system to the digestive tract wall. In some cases, an adhesive polymer may be blended together with a drug and an absorption accelerator in the drug holding layer. As a result, the system adheres to the gastrointestinal membrane wall, and good absorbability can be derived by maintaining a high concentration gradient of drug molecules between the mucosa side and inside the cell. GI-MAPS provides good absorption, but GI-MAPS is not in the category of conventional pharmaceutical preparations and is a completely new DDS, requiring a complicated manufacturing process and high manufacturing cost. There was a drawback.

  According to the present invention, the above problem can be solved by the following. In other words, if there is no water-insoluble film that forms a closed space such as GI-MAPS or an adhesive film that adheres to the small intestine, the absorption promoter and drug diffuse and dilute, and a high drug concentration gradient cannot be obtained. Absorption enhancers are also diluted and do not work effectively. Therefore, using microparticles of minute size (nanosize or micron size), the solubilized, waxy, gel or microparticle absorption promoter and peptide / protein drug, and optionally stabilizers are retained in the microparticles. Peptide, protein drug molecule, absorption enhancer, and optionally a stabilizer in the microspace while forming a high drug concentration gradient between the absorbed cells.・ A new DDS has been devised that achieves good absorption of protein drugs.

  When the peptide / protein drug is retained in the microparticles, for example, first, the microparticles are dispersed or dissolved by adding a solvent such as ethanol to a solubilizer or dispersant of the microparticles and an absorption accelerator such as a surfactant, and then the microparticles are dispersed. In addition, an absorption accelerator such as a surfactant is well permeated into the fine particles by sonication or the like. A uniform preparation can be prepared by distilling off a solvent such as ethanol under reduced pressure and then adding a peptide / protein drug or, in some cases, a stabilizer in powder or solution and stirring well at low temperature. . This production method is only one method for obtaining the solid preparation of the present invention, and is not limited to this method. The present inventor administers the obtained solid preparation into the small intestine of rats or orally as a capsule to beagle dogs, and measures peptide / protein drug concentrations in serum samples obtained over time. It was demonstrated that protein drugs are absorbed through hydrolysis in the gastrointestinal tract and then transferred into the circulating blood. The present invention has been completed based on these findings.

  That is, according to the present invention, an oral DDS (drug delivery system) of a small intestine target type for the purpose of promoting drug absorption, comprising (a) an absorption enhancer, (b) a fine particle, and (c) a peptide / protein drug. A formulation is provided.

  The small intestine target type oral DDS preparation of the present invention preferably further comprises (d) a stabilizer.

Preferably, the absorption enhancer is a liquid self-microemulsifying surfactant.
Preferably, the absorption enhancer is an ester mixture (labrazole) of glycerol ester of C6-18 fatty acid and macrogol ester of C6-18 fatty acid, polysorbate 80, monooleic acid, polyethylene glycol monooleate, polyethylene glycol monostearate , Medium chain fatty acid triglyceride, or lecithin.

  Preferably, the fine particle body is a single-walled / multi-walled carbon nanotube (for example, CNT20 manufactured by Carbon Nanotech Research Institute CNRI), carbon nanohorn (IRV Corporation), amorphous carbon nanotube (Osaka Gas Co., Ltd.) Company), Fullerene, Fullerene, Carbon 60 (SES Research, Houston, TX, USA), Charcoal fine particles (activated carbon, medicinal charcoal, pulverized activated carbon, pulverized activated carbon, etc.), porous calcium silicate (Florite-RE, Eisai Co., Ltd.) Company), porous silicic acid anhydride (SYLYSIA, Fuji Silysia Chemical), porous magnesium metasilicate aluminate (Neusilin, Fuji Chemical Industry), porous anhydrous calcium hydrogen phosphate (Fujicalin, Fuji Chemical Industry), porous calcium phosphate, Kaolin, carmellose calcium, magnesium silicate, anhydrous Lee acid, diatomaceous earth, crystalline cellulose, aluminum silicate, aluminum hydroxide, precipitated calcium carbonate, dextrin, silicon dioxide, aluminum silicate, or bentonite.

  Preferably, the stabilizer is an edible protein such as casein or lactoferrin, a natural product-derived protease inhibitor such as soybean trypsin inhibitor, or a protease inhibitor such as aprotinin.

  According to another aspect of the present invention, the method described above includes mixing (a) an absorption promoter, (b) a microparticle, and (c) a peptide / protein drug, and optionally (d) a stabilizer. The present invention provides a method for producing a small intestine target type oral DDS preparation for the purpose of promoting drug absorption.

  Peptide / protein drugs by devising a microparticulate system that delivers a high concentration gradient by delivering difficult and low-absorption drugs such as peptide / protein drugs to absorbing cells while suppressing hydrolysis by digestive enzymes A system that increases the absorbability of difficult and low absorbable drugs is provided. This system is called LFNPS (Liquid formulation holding nano-, micro-particulate system). By utilizing the LFNPS of the present invention, it becomes possible to develop peptide and protein drugs, which have been limited in clinical use as injections, as oral preparations.

Hereinafter, embodiments of the present invention will be described in detail.
The DDS preparation of the present invention comprises (a) an absorption enhancer, (b) a fine particle, and (c) a peptide / protein drug, and further comprises (d) a stabilizer in some cases. Orally administered. Hereinafter, each of these components will be described.

(1) Absorption accelerator (dissolution aid or surfactant, etc.)
Examples of absorption promoters that can be used in the present invention include surfactants, bile acids, chelators such as EDTA, organic acids, fatty acids (such as capric acid), and the like.

  The surfactant that can be used in the present invention is used to improve the bioavailability and efficacy by improving the absorption of a drug having low absorption from the gastrointestinal tract after oral administration, and exhibits a desired action. As long as the type of the surfactant is not particularly limited, a self-microemulsifying surfactant is preferable. Specific examples of surfactants that can be used in the present invention include ester mixtures of glycerol esters of C6-18 fatty acids and macrogol esters of C6-18 fatty acids (such as labrazol), polysorbate 80, monooleic acid, monoolein Examples include acid polyethylene glycol, polyethylene glycol monostearate, medium-chain fatty acid triglyceride, and lecithin.

  The glycerol ester of C6-18 fatty acid is usually used in the form of a mixture thereof as long as it contains at least one of mono-, di- and triglycerol esters of C6-18 fatty acid. The C6-18 fatty acid may be a saturated or unsaturated fatty acid having 6 to 18 carbon atoms, preferably a saturated fatty acid, particularly a saturated fatty acid having 6 to 12 carbon atoms, that is, caproic acid, caprylic acid, capric acid and lauric acid. .

  The macrogol in the macrogol ester of C6-18 fatty acid is usually polyethylene glycol having a molecular weight of 100 to 800, preferably 200 to 600, and the ester is either a mono or diester or a mono / di mixed ester. May be. The C6-18 fatty acid constituting the macrogol ester is the same as that in the aforementioned glycerol ester. The weight mixing ratio of the mixture of glycerol ester and macrogol ester is usually 1 to 0.1-10, preferably 1 to 0.2-5, for glycerol ester to macrogol ester. This mixed ester of glycerol ester and macrogol ester is a formulation additive known as a self-microemulsifying agent.

In the present invention, as a self-microemulsifying surfactant, for example, it is described as caprylocaproylmacrogolglycerides in the European Pharmacopoeia, and is marketed as a liquid labrasol (trade name) from Gattefosse SA. Can be used.
In the present invention, liquid, semi-solid or solid surfactants other than those described above can also be used. Specific examples of the surface activity that can be used in the present invention are listed below.

(A) Nonionic surfactant Alkyl glucoside, alkyl maltoside, alkylthioglucoside, lauryl macrogol glyceride, polyoxyethylene alkyl ether, polyoxyethylene alkylphenol, polyethylene glycol fatty acid ester, polyethylene glycol glycerol fatty acid ester, polyoxyethylene sorbitan Fatty acid ester, polyoxyethylene-polyoxypropylene block copolymer, polyglycerol fatty acid ester, polyoxyethylene glyceride, polyoxyethylene sterol, polyoxyethylene vegetable oil, polyoxyethylene hydrogenated vegetable oil, polyhydric alcohol and fatty acid, glyceride, vegetable oil, A reaction mixture of at least one member of the group consisting of hydrogenated vegetable oils and sterols, sucrose esters, Sugar ethers, sucrose glycerides, or mixtures thereof.

(B) Hydrophilic surfactant
PEG-10 laurate, PEG-12 laurate, PEG-20 laurate, PEG-32 laurate, PEG-32 dilaurate, PEG-12 oleate, PEG-15 oleate, PEG -20 oleate, PEG-20 dioleate, PEG-32 oleate, PEG-200 oleate, PEG-400 oleate, PEG-15 stearate, PEG-32 distearate, PEG- 40 Stearic acid ester, PEG-100 stearic acid ester, PEG-20 dilauric acid ester, PEG-25 glyceryl trioleic acid ester, PEG-32 dioleic acid ester, PEG-20 glyceryl lauric acid ester, PEG-30 glyceryl lauric acid ester PEG-20 glyceryl stearate, PEG-20 glyceryl oleate, PEG-30 glyceryl oleate, PEG-30 glyceryl laurate, PEG-40 glyceryl laurate, PEG-40 palm kernel oil, PEG-50 hydrogenated castor oil, PEG-40 castor oil, PEG-35 castor oil, PEG-60 castor oil, PEG -40 hydrogenated castor oil, PEG-60 hydrogenated castor oil, PEG-60 corn oil, PEG-6 caprate / caprylate glyceride, PEG-8 caprate / caprylate glyceride, polyglyceryl-10 laurate, PEG-30 cholesterol PEG-25 phytosterol, PEG-30 soybean sterol, PEG-20 trioleate, PEG-40 sorbitan oleate, PEG-80 sorbitan laurate, polysorbate 20, polysorbate 80, POE-9 lauryl ether, POE- 23 Lauryl ether, POE-10 oleyl ether, POE-20 oleyl ether, POE-20 stearyl ether, tocopheryl PEG-100 succinate, PE G-24 cholesterol, polyglyceryl-10 oleate, Tween 40, Tween 60, sucrose monostearate, sucrose monolaurate, sucrose monopalmitate, PEG 10-100 nonylphenol, PEG 15-100 octylphenol, poloxamer , And mixtures thereof.

(C) Ionic surfactants: alkyl ammonium salts; bile salts; fusidic acid; fatty acid conjugates of amino acids, oligopeptides and polypeptides; glyceride esters of amino acids, oligopeptides and polypeptides; acyl lactylates; mono and diglyceride mono Succinylated monoglycerides; citrate esters of mono and diglycerides; alginate; propylene glycol alginate; lecithin and hydrogenated lecithin; lysolecithin and hydrogenated lysolecithin; lysophospholipid; camitin fatty acid ester salt; phospholipid; Sulfate salts; fatty acid salts; docusate sodium; and salts thereof.
Specific examples of the ionic surfactant include the following.

  Lecithin, lysolecithin, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidic acid, phosphatidylserine, lysophosphatidylcholine, lysophosphatidylethanolamine, lysophosphatidylglycerol, lysophosphatidic acid, lysophosphatidylserine, PEG phosphatidylethanolamine, PVP fatty acid Lactylate, stearoyl-2-lactylate, succinylated monoglyceride, mono / diglyceride mono / diacetylated tartaric acid ester, mono / diglyceride citrate ester, cholic acid ester, taurocholic acid ester, glycocholic acid ester, deoxycholic acid Ester, Taurodeoxycholic acid ester, Kenodeki Cholic acid ester, glycodeoxycholic acid ester, glycochenodeoxycholic acid ester, taurochenodeoxycholic acid ester, ursodeoxycholic acid ester, lithocholic acid ester, tauroursodeoxycholic acid ester, glycoursodeoxycholic acid ester, corrisarcosine, N-methyl Taurocholate, caproate, caprylate, caprate, laurate, myristate, palmitate, oleate, ricinolate, linoleate, linoleate, stearate, lauryl sulfate, tetraacetyl sulfate, docusate, lauroyl carnitine, palmitoyl carnitine, myristoyl carnitine, and their Salts and mixtures thereof.

(D) Hydrophobic surfactant Alcohol; Polyoxyethylene alkyl ether; Fatty acid; Bile salt; Glycerol fatty acid ester; Acetylated glycerol fatty acid ester; Lower alcohol fatty acid ester; Polyethylene glycol fatty acid ester; Polyethylene glycol glycerol fatty acid ester; Fatty acid ester; Polyoxyethylene glyceride; Lactate ester of mono / diglyceride; Propylene glycol diglyceride; Sorbitan fatty acid ester; Polyoxyethylene sorbitan fatty acid ester; Polyoxyethylene-polyoxypropylene block copolymer; Transesterified vegetable oil; Sterol; Sugar ester Sugar ether; sucrose glyceride; polyoxyethylene vegetable oil; polyoxyethylene hydrogenated vegetable oil; Triol and fatty acid, glycerides, the reaction mixture of at least one of the group consisting of vegetable oils, hydrogenated vegetable oils and sterols; and mixtures thereof.

Specific examples of the hydrophobic surfactant include the following.
Oleic acid; lauric acid; stearic acid; palmitic acid; PEG 1-4 stearate; PEG 2-4 oleate; PEG-4 dilaurate; PEG-4 dioleate; PEG-4 distearate PEG-6 dioleate; PEG-8 dioleate; PEG 3-16 castor oil; PEG 5-10 hydrogenated castor oil; PEG 6-20 corn oil; PEG 6-20 Almond oil; PEG-6 olive oil; PEG-6 peanut oil; PEG-6 palm kernel oil; PEG-6 hydrogenated palm kernel oil; PEG-4 capric / capric glyceride mono-, di-, tri-, and tetra-esters of vegetable oil and sorbitol ; Pentaerythrityl di, tetrastearic acid ester, isostearic acid ester; oleic acid ester; caprylic acid ester or capric acid ester; polyglyceryl 2-4 oleic acid ester Tellurium, stearate or isostearate; polyglyceryl 4-10 pentaoleate; polyglyceryl-3 dioleate; polyglyceryl-6 dioleate; polyglyceryl-10 trioleate; polyglyceryl-3 distearate; C. C.sub.6 to C.sub.22 fatty acid propylene glycol mono or diester; C.sub.6 to C.sub.22 fatty acid monoglyceride; C.sub.6 to C.sub.22 fatty acid acetylated monoglyceride; C. 6 to C.22 fatty acid diglycerides; lactic acid esters of monoglycerides; lactic acid esters of diglycerides; cholesterol; phytosterols; PEG 5-20 soy sterols; PEG-6 sorbitan tetras, hexastearic acid esters; PEG-6 sorbitans Tetraoleic acid ester; sorbitan monolauric acid ester; Rubitan monopalmitate; sorbitan mono, trioleate; sorbitan mono, tristearate; sorbitan monoisostearate; sorbitan sesqueoleate; sorbitan sesquiterate; PEG 2-5 oleyl ether; POE 2 -4 lauryl ether; PEG-2 cetyl ether; PEG-2 stearyl ether; sucrose distearate; sucrose dipalmitate; ethyl oleate; isopropyl myristate; isopropyl palmitate; ethyl linolenate; isopropyl linoleate; poloxamer; Cholic acid; ursodeoxycholic acid; glycocholic acid; taurocholic acid; lysocholic acid; deoxycholic acid; chenodeoxycholic acid; and mixtures thereof.

The following are mentioned as a further specific example of the absorption promoter which can be used by this invention.
Water-immiscible triglyceride vegetable oils (safflower oil, sesame oil, corn oil, castor oil, coconut oil, cottonseed oil, soybean oil, olive oil, etc .; water-immiscible refined and synthetic and semi-synthetic oils (eg mineral oil), known as MIGLYOL.RTM Triglycerides (caprylic / capric triglycerides, caprylic / capric / linoleic triglycerides, long chain triglycerides such as triolein, other mixed chain triglycerides that are liquid at room temperature, monoglycerides, diglycerides, and mono, di, Fatty acids and esters; water-soluble alcohols, glycerin and propylene glycol; water-miscible polyethylene glycols that are liquid at room temperature, such as PEG-400.

  Commercially available products include corn oil, propylene glycol, CREMOPHOR RH-40 (polyoxy-40 hydrogenated castor oil), LABRAFIL M 2125 (linoleoyl polyoxy-6 glyceride) and 1944 (oleoyl polyoxy-6 glyceride), ethanol , PEG 400, Polysorbate 80, glycerin, peppermint oil, soybean oil (long chain triglyceride), sesame oil (long chain triglyceride), propylene carbonate, and tocopheroyl TPGS, MIGLYOL 812 (caprylic acid / capric acid triglyceride), oleic acid, olive oil ( Long chain triglycerides), CAPMUL MCM (medium chain monoglycerides), CAPMUL PG-8 (propylene glycol caprylyl mono and diglycerides), CREMOPHOR EL (polyoxy 35 castor oil), LABRASOL (caprylocaproyl polyoxy-8 glycerides), triacetin ( Acetyl triglyceride), MAISINE 35-1 (glyceryl monolinolei) Acid), OLICINE (glyceryl monooleate / linoleate), PECEOL (glyceryl monooleate), TRANSCUTOL P (diethylene glycol monoethyl ether), PLUROL Oleique CC (polyglyceryl-6 dioleate), LAUROGLYCOL 90 (propylene) Glycol monolaurate), CAPRYOL 90 (propylene glycol monocaprylic acid), MYVACETS (acetylated monoglyceride), ARLACELS (sorbitan fatty acid ester), PLURONICS (propylene and ethylene oxide copolymer), BRIJ 30 (polyoxyethylene 4-lauryl ether), GELUCIRE 44/14 (lauroyl polyoxyl-32 glyceride), GELUCIRE 33/01 (glycerol ester of fatty acid) and the like.

  Other specific examples of commercially available surfactants include benzethonium chloride (HYAMINE.RTM. 1622, Lonza, Inc., Fairlawn, NJ); DOCUSATE SODIUM (Mallinckrodt Spec. Chem., St. Louis, Mo.); Oxyethylenesorbitan fatty acid esters (TWEEN.RTM., ICI Americas Inc., Wilmington, Del.); LIPOSORB.RTM. P-20 (Lipochem Inc., Patterson NJ); CAPMUL.RTM. POE-0 (Abitec Corp., Janesville, Wis.).

  Further examples of absorption enhancers that can be used in the present invention include fatty acids such as capric acid and its derivatives such as N- [8- (2-hydroxybenzoyl) amino] caprylic acid (SNAC) and Sodium N- [8- ( 2-hydroxybenzoyl) amino] decanate (SNAD), amino acid enamine derivatives (such as ethylglycol acetoacetate of phenylglycine), sodium salicylate or its derivatives, mixed micelles (mixed micelles of monoolein and sodium glycocholate or sodium taurocholate, etc.) ), N-acyl collagen peptide, sodium N-acyl amino acid, life-spanning saponin and the like.

(2) Fine Particles Examples of fine particles that can be used in the present invention include single-walled and multi-walled carbon nanotubes (for example, CNT20 manufactured by Carbon Nanotech Research Institute CNRI), carbon nanohorns (IRV shares) Company), Fullerene, Fullerene, Carbon 60 (SES Research, Houston, TX, USA), amorphous carbon nanotubes (Osaka Gas Co., Ltd.), charcoal fine particles (medicine charcoal, coconut husk activated carbon, pulverized coconut husk activated carbon, etc.) Carbon compounds, porous calcium silicate (Florite-RE, Eisai Co., Ltd.), porous anhydrous silicic acid (SYLYSIA, Fuji Silysia Chemical), porous magnesium metasilicate aluminate (Neusilin, Fuji Chemical Industry), porous anhydrous Calcium hydrogen phosphate (Fujicalin, Fuji Chemical), porous calcium phosphate , Kaolin, carmellose calcium, magnesium silicate, silicic anhydride, diatomaceous earth, crystalline cellulose, aluminum silicate, aluminum hydroxide, precipitated calcium carbonate, dextrin, silicon dioxide, aluminum silicate, bentonite, and other porous materials Is mentioned.

  In the present invention, the mixing ratio of the fine particles and the absorption accelerator is not particularly limited, but is generally 1: 0.5 to 1: 100, and preferably 1: 1 to 1:50.

(3) Peptide / protein drugs Examples of the present invention are given for erythropoietin, interferon, etc., but the application of this patent is not limited to these peptide / protein drugs, and is applicable to a wide range of peptide / protein drugs. It is possible DDS technology.
The content of the active ingredient drug in the oral DDS preparation of the present invention is not particularly limited, but is generally about 0.1 to 50% by weight.

(4) Stabilizer When a peptide / protein drug is susceptible to hydrolysis by digestive enzymes, a better absorbability of the peptide / protein drug can be obtained by adding a stabilizer. For example, edible proteins such as casein and lactoferrin, proteolytic enzyme inhibitors derived from natural products such as soybean trypsin inhibitor, and proteolytic enzyme inhibitors such as aprotinin are typical stabilizers. However, it is not limited to these stabilizers.

(5) Formulation and Formulation In the oral DDS preparation of the present invention, the absorption accelerator and the fine particle are mixed and solidified, and then added with a peptide or protein drug in a powder or solution state and a stabilizer as required. It can be produced by mixing well. It can also be produced by mixing and solidifying a solution containing an absorption promoter, a peptide / protein drug, and optionally a stabilizer, and fine particles. In this case, the peptide / protein drug is mixed with the microparticles separately from the absorption promoter or as a mixture. When preparing a solution containing an absorption enhancer and a peptide / protein drug, the peptide / protein drug can be dissolved in an appropriate solvent such as purified water or a buffer and then mixed with the absorption enhancer.

Alternatively, the preparation solidified as described above can be directly administered to a patient, but can be administered to a patient after being prepared into various preparation forms known in the preparation field. That is, the oral DDS preparation of the present invention is appropriately prepared by using conventional additives such as pharmaceutically acceptable excipients, carriers and diluents, capsules, tablets, powders, granules, fine granules, It can be used as a preparation such as a pill or a powder preparation. For example, in the case of production of solid preparations such as capsules, tablets, powders, or granules, as necessary, excipients such as lactose, glucose, sucrose, and mannitol, disintegrants such as starch and sodium alginate, Lubricants such as magnesium stearate and talc, binders such as polyvinyl alcohol and hydroxypropyl cellulose can be used.
The present invention will be described more specifically with reference to the following examples, but the present invention is not limited to the examples.

Example 1
Add about 5 ml of ethanol to 100 mg of polyvinylpyrrolidone K-90, 1.25 g of Labrasol, and 125 mg of carbon nanotube CNT20 (CNRI), vibrate for 15 minutes under ultrasound, and then stir for 1 hour with a stir bar. To do. The ethanol is removed by stirring overnight on a stirrer while heating to about 50 ° C. After cooling, add 52 μl (equivalent to 2500 IU) of erythropoietin injection solution (for Espoo ^TM subcutaneous use, 24000 IU / 0.5 ml) and knead well to obtain a solid preparation.

(Evaluation)
Wistar male rats weighing approximately 400 g were subjected to laparotomy under pentobarbital anesthesia and blank blood was collected from the jugular vein, and then 24 mg of the preparation of Example 1 was administered into the ileum per 400 g body weight. Thereafter, blood was collected from the jugular vein for 6 hours, and the serum EPO concentration was measured by ELISA using an assay kit (EPO ELISA) from Roche Diagnostics GmbH (Manhaim, Germany). As a control preparation for comparison, an EPO solution and a solution obtained by adding Labrasol to the EPO solution were used. The EPO solution was orally administered to rats using a sonde at a dose of 100 IU / kg. Labrasol-added EPO solution was injected at the same dose into the ileum of rats. The uppermost value in Table 1 below represents the time course of the average value of the plasma EPO concentration in 4 rats of the preparation of Example 1. The middle row shows the value after adding Labrasol to the EPO solution, and the lower row shows the value after administration of the EPO solution.

Example 2
Add about 5 ml of ethanol to 100 mg of Labrasol and 10 mg of carbon nanotube CNT20 (CNRI), vibrate for 15 minutes under ultrasonic waves, and then stir for 1 hour with a stirrer bar. The ethanol is removed by stirring overnight on a stirrer while heating to about 50 ° C. After cooling, add 4.16 μl (equivalent to 200 IU) of erythropoietin injection solution (espo ^TM subcutaneous use, 24000 IU / 0.5 ml) and knead well to obtain a solid preparation.

Example 3
Add about 5 ml of ethanol to 100 mg of Labrasol and 10 mg of carbon nanotube CNT20 (CNRI), vibrate for 15 minutes under ultrasonic waves, and then stir for 1 hour with a stirrer bar. The ethanol is removed by stirring overnight on a stirrer while heating to about 50 ° C. After cooling, add about 4.16 μl (equivalent to 200 IU) of erythropoietin injection solution (for Espoo ^™ subcutaneous use, 24000 IU / 0.5 ml) and 100 mg of casein, and knead well to obtain a solid preparation.

Example 4
To 100 mg of Labrasol, add 10 mg of carbon nanotube CNT20 (CNRI) and 5 mg of sodium carboxymethyl starch (trade name Explotab) and mix well. Add 4.16 μl (equivalent to 200 IU) of erythropoietin injection solution (espo TM subcutaneous use, 24000 IU / 0.5 ml) and 50 mg of lactoferrin from bovine milk (Wako Pure Chemical) from milk, and mix well to obtain a solid preparation.

(Evaluation)
Evaluation was performed at a dose of 100 IU / kg of EPO using rats in the same manner as in Example 1. The upper value in Table 2 below represents the time course of the average value of serum EPO concentration in 4 rats of the preparation of Example 2. The middle row shows the time course of the mean serum EPO concentration in 4 rats of the preparation of Example 3. The lower row shows the results of Example 4. The addition of casein, a stabilizer, resulted in higher serum EPO concentrations.

Example 5
Thoroughly mix 100 mg of Labrasol and 60 mg of silicia, add 2.08 μl (equivalent to 100 IU) of erythropoietin injection solution (Espoe TM subcutaneous, 24000 IU / 0.5 ml), and knead well to obtain a solid preparation.

(Evaluation)
Wistar male rats were administered into the stomach or jejunum at an EPO dose of 100 IU / kg. Thereafter, circulating blood was collected over time, and serum EPO concentration was measured. The upper part of Table 3 below shows the results of intragastric administration, and the lower part shows the results of intraileal administration.

Example 6
Add about 5 ml of ethanol to 100 mg of polyvinylpyrrolidone K-90, 1.25 g of Labrasol, and 125 mg of carbon nanotube CNT20 (CNRI), vibrate for 15 minutes under ultrasound, and then stir for 1 hour with a stir bar. To do. The ethanol is removed by stirring overnight on a stirrer while heating to about 50 ° C. After cooling, add 52.1 μl (equivalent to 2500 IU) of erythropoietin injection solution (espo ^TM subcutaneous use, 24000 IU / 0.5 ml) and knead well to obtain a solid preparation. After weighing 600 mg of a solid preparation and wrapping it with a commercially available wafer, it is placed in an enteric capsule prepared by Eudragit S100 to obtain an oral preparation.

Example 7
A solid preparation prepared in the same manner as in Example 6 is placed in an enteric capsule prepared with Eudragit L100 to obtain an oral preparation.

Example 8
Add about 5 ml of ethanol to 900 mg of Silicia 550 and 1.5 g of Labrasol, vibrate under ultrasonic waves for 15 minutes, add a stir bar and stir for 1 hour. The ethanol is removed by stirring overnight on a stirrer while heating to about 50 ° C. After cooling, add 20.8 μl (equivalent to 1000 IU) of erythropoietin injection solution (for Espoo ^TM subcutaneous use, 24000 IU / 0.5 ml) to 800 mg of the mixture, and knead well to obtain a solid preparation. After weighing 820.8 mg of a solid preparation and wrapping it with a commercially available wafer, it is placed in an enteric capsule made of Eudragit L100 to give an oral preparation.

(Evaluation of formulations of Examples 6 to 8)
After blank blood was collected from the jugular vein of a male beagle dog (body weight: about 10 kg), the preparations of Examples 6 to 8 prepared above were orally administered at 100 IU / kg as an EPO dose, and thereafter 8 hours every hour. Blood was collected from the jugular vein until time, and the EPO concentration in plasma was measured by ELISA using an assay kit (EPO ELISA) from Roche Diagnostics GmbH (Manhaim, Germany). Table 4 below shows the time course of the mean value of plasma EPO concentration in 3 beagle dogs. The upper part shows the value of the preparation of Example 6, the middle part shows the preparation of Example 7, and the lower part shows the value of the preparation of Example 8.

Example 9
Add about 5 ml of ethanol to 1.0 g of Labrasol and 0.5 g of medicinal charcoal, shake for 15 minutes under ultrasonic waves, add a stir bar and stir for 1 hour. The ethanol is removed by stirring overnight on a stirrer while heating to about 50 ° C. Add 2.08 μl (equivalent to 100 IU) of erythropoietin injection solution (for Espoo ^TM subcutaneous use, 24000 IU / 0.5 ml) to 130 mg of the resulting fine particles, and knead well to obtain a solid preparation. 132 mg of solid preparation per kg body weight of the rat was administered into the ileum. Table 5 below shows the time course of the mean value of serum EPO concentration in 3 rats.

Example 10
Add about 5 ml of ethanol to 1.0 g of Labrasol and 1160 mg of finely ground coconut shell activated carbon, vibrate under ultrasonic waves for 15 minutes, add a stirrer bar and stir for 1 hour. The ethanol is removed by stirring overnight on a stirrer while heating to about 50 ° C. To 108 mg of the obtained fine particles, 4.16 μl (equivalent to 200 IU) of erythropoietin injection solution (Espoe ^TM subcutaneous use, 24000 IU / 0.5 ml) is added and kneaded well to obtain a solid preparation. A solid preparation of 112 mg / kg body weight of the rat is administered into the jejunum of the rat. Table 6 below shows the time course of the mean value of plasma EPO concentration in 3 rats.

Example 11
Add 200 mg of Labrasol to 75 mg of carbon nanohorn (IR Buoy Co., Ltd.) and mix well to obtain an admixture. Add 2.08 μl (equivalent to 100 IU) of erythropoietin injection solution (espo ^TM subcutaneous use, 24000 IU / 0.5 ml) to 68.8 mg of the mixture, and knead well to obtain a solid preparation. The solid preparation was administered into the jejunum at 70.88 mg / kg body weight of the rat. Table 7 below shows the time course of the average value of serum EPO concentration in 3 rats.

Example 12
About 5 ml of ethanol is added to 50 mg of Labrasol and 150 mg of fullerene Fullerene, Carbon 60 (manufactured by SES Research, Houston, TX, USA), and the mixture is vibrated for 15 minutes under ultrasonic waves. The ethanol is removed by stirring overnight on a stirrer while heating to about 50 ° C. Add 2.08 μl (equivalent to 100 IU) of erythropoietin injection solution (espo ^TM subcutaneous use, 24000 IU / 0.5 ml) to the fine particles obtained and knead well to obtain a solid preparation. 80.8 mg of solid agent is administered into the jejunum of a rat weighing about 400 g. Table 8 below shows the time course of mean serum EPO concentration in 3 rats.

Example 13
Add about 5 ml of ethanol to 80 mg of polyvinylpyrrolidone K-90, 1.0 g of Labrasol, and 100 mg of carbon nanotube CNT20, vibrate for 15 minutes under ultrasonic waves, add a stirrer bar and stir for 1 hour. The ethanol is removed by stirring overnight on a stirrer while heating to about 50 ° C. Add 16.6 μl (equivalent to 100,000 IU) of an interferon injection solution (Sumiferon ^™ , 6 million IU / ml) to 122 mg of the fine particle mixture, and knead well.

(Evaluation)
Wistar male rats weighing approximately 400 g were subjected to laparotomy under pentobarbital anesthesia and blank blood was collected from the jugular vein, and then 27.7 mg of the formulation of Example 13 per 400 g body weight was administered into the ileum. Thereafter, blood was collected from the jugular vein for 6 hours, and the interferon concentration in the serum was measured by ELISA using an assay kit (Human IFNα ELISA kit) of Biosource International (CA, USA). Table 9 below shows the time course of serum interferon concentration (average value of 3 to 4 cases) (unit: IU / ml).

Example 14
Add 40 mg of carbon nanotube CNT20 to 270 mg of Gercia Gelucire 44/14 and 20 mg of Pharmasol Pharmasol, and mix well with a micro spatula while heating to about 50 ° C. After cooling, 16.6 μl (equivalent to 100,000 IU) of an interferon injection solution (Sumiferon ^™ , 6 million IU / ml) is added to 123 mg of this mixture of microparticles and kneaded well to obtain a solid preparation.

Example 15
Add 40 mg of carbon nanotube CNT20 to 270 mg of Gercia Gelucire 44/14 and 20 mg of Pharmasol Pharmasol, and mix well with a micro spatula while heating to about 50 ° C. After cooling, 16.6 μl of interferon injection solution (Sumiferon TM, 6 million IU / ml) and 50 mg of lactoferrin and 123 mg of this mixture of microparticles and kneaded well to obtain a solid preparation

Example 16
Add 40 mg of carbon nanotube CNT20 to 270 mg of Gercia Gelucire 44/14 and 20 mg of Pharmasol Pharmasol, and mix well with a micro spatula while heating to about 50 ° C. While maintaining at about 40 ° C., add 100 mg of casein and mix well. After cooling, 8.3 μl (equivalent to 50,000 IU) of an interferon injection solution (Sumiferon ^™ , 6 million IU / ml) is added to 80 mg of this mixture of microparticles and kneaded well to obtain a solid preparation.

(Evaluation)
Wistar male rats weighing about 400 g were subjected to laparotomy under pentobarbital anesthesia, and blank blood was collected from the jugular vein, and then the formulations of Examples 14, 15 and 16 were administered at a dose of about 30 mg per 400 g body weight as interferon. Each group was administered into the ileum of a different group of rats so as to be 50,000 IU / kg. Thereafter, blood was collected from the jugular vein for 6 hours, and the interferon concentration in the serum was measured by ELISA using an assay kit (Human IFNα ELISA kit) of Biosource International (CA, USA). Table 10 below shows the time course of serum interferon concentration (average value of 3 to 4 cases) (unit: IU / ml). The upper row shows the evaluation results of the preparation of Example 14, the middle row shows the evaluation results of Example 15, and the lower row shows the evaluation results of the preparation of Example 16.

Example 17
Add about 5 ml of ethanol to 80 mg of polyvinylpyrrolidone K-90, 1.0 g of Labrasol, and 100 mg of carbon nanotube CNT20, vibrate for 15 minutes under ultrasonic waves, add a stirrer bar and stir for 1 hour. The ethanol is removed by stirring overnight on a stirrer while heating to about 50 ° C. After cooling, 8.5 μl of peginterferon-α-2a (Pegasys ^™ , Chugai Pharmaceutical Co., Ltd.) is added to 59 mg of the mixture of fine particles and kneaded well to obtain a solid preparation.

(Evaluation)
A Wistar male rat weighing about 400 g was subjected to laparotomy under pentobarbital anesthesia and blank blood was collected from the jugular vein, and then 27 mg of the preparation of Example 17 per 400 g body weight was administered into the jejunum. Thereafter, blood was collected from the jugular vein for 6 hours, and the peginterferon-α-2a concentration in the serum was measured by ELISA using an assay kit (Human IFNα ELISA kit) of Biosource International (CA, USA). Table 11 below shows the time course of average serum peginterferon-α-2a concentration (average value of 4 rats) (unit: IU / ml).

Example 18
About 80 ml of ethanol is added to 80 mg of polyvinylpyrrolidone K-90, 1000 mg of Labrasol and 100 mg of carbon nanotube CNT20, and the mixture is vibrated for 15 minutes under ultrasonic waves, and then stirred for 1 hour with a stirrer bar. The ethanol is removed by stirring overnight on a stirrer while heating to about 50 ° C. Dissolve 1.0 mg of insulin sodium salt (Aldrich) in 0.1 ml of pH 7.4 phosphate buffer. Add 80 μl of insulin solution to 118 mg of carbon nanotube blend brought to room temperature and mix. Water is distilled off under reduced pressure to obtain a solid agent.

(Evaluation) Rat Absorption Experiment A Wistar male rat having a body weight of about 400 g is fixed to an operating table under pentobarbital anesthesia. After collecting blank blood from the jugular vein, the above preparation of Example 18 was administered into the ileum. Thereafter, blood was collected from the jugular vein after 30 minutes, 1 hour, 1.5 hours, 2 hours, 3 hours, 4 hours, and 5 hours, and the serum glucose concentration was measured using Glucose B-Test Wako (Wako Pure Chemical Industries). . Table 12 below shows the time course of the serum glucose concentration, that is, the average value of blood glucose level (mg / dl) in three rats.

Example 19
Add 100 mg of Labrasol 36 mg of amorphous carbon nanotubes (manufactured by Osaka Gas Co., Ltd.) and 5 mg of sodium carboxymethyl starch (trade name Explotab) and mix well. Add erythropoietin injection solution (espo ^TM subcutaneous use, 24000 IU / 0.5 ml) of 4.16 μl (equivalent to 200 IU) and 50 mg of casein Casein (Wako Pure Chemical Industries, Ltd.) and knead well to obtain a solid preparation. Rats were administered into the jejunum at an EPO dose of 100 IU / kg. Thereafter, the change in circulating serum EPO concentration was measured by ELISA. The measurement results are shown in Table 13 below.

  Many oral DDS technologies have been studied to improve the absorption of peptide and protein drugs, but unfortunately they have not been commercialized. The reason for this is that after oral administration and after release from the preparation in the digestive tract, the absorption enhancer is separated and diluted from difficult / low absorbable drugs such as peptide / protein drugs, and the peptide / protein drugs are hydrolyzed. Because. Therefore, LFNPS was invented as a system that increases the absorbability of drugs by delivering absorption promoters and peptide / protein drugs to the absorbing cells while preventing hydrolysis by digestive enzymes to form a high concentration gradient. According to the present invention, it has become possible to make a peptide / protein drug, which has heretofore been limited in clinical use as an injection, into an oral preparation.

Claims (7)

A small intestine target type oral DDS preparation for the purpose of promoting drug absorption, comprising (a) an absorption enhancer, (b) a fine particle, and (c) a peptide / protein drug. The small intestine target type oral DDS preparation for the purpose of promoting drug absorption according to claim 1, further comprising (d) a stabilizer. 3. The small intestine target type oral DDS preparation for the purpose of promoting drug absorption of a drug according to claim 1 or 2, wherein the absorption enhancer is a liquid self-microemulsifying surfactant. Absorption enhancer is ester mixture (labrazole) of glycerol ester of C6-18 fatty acid and macrogol ester of C6-18 fatty acid, polysorbate 80, monooleic acid, polyethylene glycol monooleate, polyethylene glycol monostearate, medium chain The small intestine target type oral DDS preparation for the purpose of promoting drug absorption according to claim 1, which is fatty acid triglyceride or lecithin. The fine particles are single-walled / multi-walled carbon nanotubes (for example, CNT20 manufactured by Carbon Nanotech Research Institute, Inc.), carbon nanohorn (IRV Corporation), amorphous carbon nanotube (Osaka Gas Co., Ltd.), Fullerene, Carbon 60 (manufactured by SES Research, Houston, TX, USA), charcoal fine particles (activated carbon, medicinal charcoal, pulverized activated carbon, pulverized coconut shell activated carbon, etc.), porous calcium silicate (Florite-RE, Eisai Co., Ltd.), Porous anhydrous silicic acid (SYLYSIA, Fuji Silysia Chemical), porous magnesium metasilicate aluminate (Neusilin, Fuji Chemical Industry), porous anhydrous calcium hydrogen phosphate (Fujicalin, Fuji Chemical Industry), porous calcium phosphate, kaolin, carme Loin calcium, magnesium silicate, anhydrous silicic acid, silicic acid The object is to promote drug absorption according to any one of claims 1 to 4, which is a clay, crystalline cellulose, aluminum silicate, aluminum hydroxide, precipitated calcium carbonate, dextrin, silicon dioxide, aluminum silicate, or bentonite. Small intestine target type oral DDS formulation. 6. The stabilizer according to claim 2, wherein the stabilizer is an edible protein such as casein or lactoferrin, a proteolytic enzyme inhibitor derived from a natural product such as soybean trypsin inhibitor, or a proteolytic enzyme inhibitor such as aprotinin. A small intestine target type oral DDS preparation for the purpose of promoting drug absorption. The drug according to any one of claims 1 to 6, which comprises mixing (a) an absorption accelerator, (b) a microparticle, and (c) a peptide / protein drug and optionally (d) a stabilizer. A method for producing a small intestine target type oral DDS preparation for the purpose of promoting absorption.