MXPA01008326A - - Google Patents

Description

LIPOSOMAL PREPARATIONS TECHNICAL FIELD This invention relates to a pharmaceutical liposomal preparation comprising, as an active ingredient, a pipecolic acid derivative which is of particular interest lately because of its excellent immunosuppressive activity, particularly a macrolide compound, for example a tricyclic compound known as tacrolimus (FK506) or a pharmaceutically acceptable salt thereof. More particularly, the present invention relates to a liposomal preparation comprising the aforementioned active ingredient, entrapped in liposomes and consequently capable of maintaining the solution stable in various media such as physiological saline solution, glucose solution for injection, water or juice and, therefore, that is applicable to various methods of administration including injections such as intravenous injection, intramuscular injection, and topical injections for intra-articular administration and the like, topical administration such as application to the skin, instillation in the eye, nasal administration, and inhalation, and in addition, oral administration and rectal administration, etc. In particular, the present invention relates to a liposomal preparation characterized by not containing cholesterol and because it has a rapid action for diseases such as cerebral ischemia by bolus administration.

BACKGROUND OF THE INVENTION As a liposomal preparation containing tacrolimus, for example, those prepared by incorporating a stabilizer such as cholesterol into a phospholipid, as a main ingredient for the formation of liposome (O93 / 08802) have been known. With such a constitution, it becomes possible to prepare a liquid preparation from tacrolimus, which is slightly soluble in water. Even if such a preparation is contacted with a body fluid, the crystallization of an active ingredient does not occur, so that the preparation shows excellent bioavailability and is stable. Therefore, the preparation can take any dosage form represented by injection, instillation in the eye, nasal administration, inhalation, percutaneous absorbent, topical injection and the like. In addition, it is also possible to increase the intense transmigration of tacrolimus to a site where the transmigration of tacrolimus is particularly desired, and to suppress its transmigration to a site where transmigration is not necessarily desired. It is known that excellent effects in practice, such as increased drug efficacy, reduction of side effects and persistence of drug efficacy, are obtained as a result. The liposomal preparation shows an excellent effect for the treatment of cerebral ischemic diseases such as cerebral infarction. However, since a liposomal membrane is not stable, the liposomal preparation does not show sufficient rapid action, such as an anticoagulant, a fibrinolytic agent and a cerebrovascular dilator used as a medicinal treatment for cerebral infarction. Therefore, it has been required to develop a drug that has an excellent fast action capable of coping with an emergent situation such as cerebral infarction. An object of the present invention is to improve the problems described above, thereby providing a liposomal preparation containing the pipecolic acid derivative, which has an excellent fast action.

DESCRIPTION OF THE INVENTION The present invention is directed to a liposomal preparation comprising, as an active ingredient, a pipecolic acid derivative represented by a macrolide compound of the following general formula (I) or a pharmaceutically acceptable salt thereof trapped in a liposome. The pipecolic acid derivatives in the present invention mean those which have a common activity, capable of having an affinity for the FKBP type immunophilin and which inhibit peptidyl-proryl-isomerase and / or the activity of the rotamase enzyme, and which have a common chemical structure, capable of being derived from pipecolic acid. A specific example of the pipecolic acid derivatives includes a macrolide compound such as the tricyclic compound of the following general formula (I) or a pharmaceutically acceptable salt thereof: wherein each of the adjacent pairs of R1 and R2, R3 and R4, and R5 and R6 independently: a) are two adjacent hydrogen atoms; or b) they can form another bond formed between the carbon atoms to which they are attached; and R2 may also be an alkyl group; R7 is a hydrogen atom, a hydroxyl group, a protected hydroxyl group, or an alkoxy group, or an oxo group together with R1; R8 and R9 are independently a hydrogen atom or a hydroxyl group; R10 is a hydrogen atom, an alkyl group, an alkyl group substituted with one or more hydroxyl groups, an alkenyl group, an alkenyl group substituted with one or more hydroxyl groups, or an alkyl group substituted with an oxo group; X is an oxo group, a state where a hydrogen atom and a hydroxyl group are attached to a carbon atom, a state where two hydrogen atoms are attached to a carbon atom, or a group represented by the formula -CH20-; And it is an oxo group, a state where a hydrogen atom and a hydroxyl group are attached to a carbon atom, a state where two hydrogen atoms are attached to a carbon atom, or a group represented by the formula = N- NR11R12 or N-OR13; R11 and R12 are independently a hydrogen atom, an alkyl group, an aryl group, or a tosyl group; R13, R14, R15, R16, R17, R18, R19, R22 and R23 are independently a hydrogen atom or an alkyl group; R24 is an optionally substituted heterocyclic ring which may contain one or more heteroatoms; and n represents an integer of 1 or 2. In addition to the above definitions, Y, R10 and R23, together with the carbon atoms to which they are attached, may represent a saturated or unsaturated 5 or 6 membered heterocyclic ring containing nitrogen, sulfur and / or oxygen atoms, and the heterocyclic ring may be substituted with one or more groups selected from the group consisting of an alkyl group, a hydroxyl group, an alkyloxy group, a benzyl group, a group of the formula -CH2Se (C6H5), and an alkyl substituted with one or more hydroxyl groups. The definitions used in the general formula (I) and the specific and preferred examples thereof are explained and described in detail. The term "lower" means, unless stated otherwise, a group having 1 to 6 carbon atoms. Preferable examples of the "alkyl group" and an alkyl portion of the "alkoxy group" include a straight or branched chain aliphatic hydrocarbon residue, for example, a lower alkyl group such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, neopentyl and hexyl. Preferable examples of the group "alkenyl group" include a straight or branched chain aliphatic hydrocarbon residue having a double bond, for example, a lower alkenyl group such as vinyl, propenyl (for example, allyl), butenyl, methylpropenyl, pentenyl and hexenyl. Preferable examples of the "aryl group" include phenyl, tolyl, xylyl, cumenyl, mesityl and naphthyl. Preferred protective groups in the "protected hydroxyl groups" and in a "protected amino" described below include the group 1- (lower alkylthio) (lower alkyl) such as a lower alkylthiomethyl group (for example, methylthiomethyl, ethylthiomethyl, propylthiomethyl, isopropylthiomethyl) , butylthiomethyl, isobutylthiomethyl, hexylthiomethyl, etc.), more preferably the alkylthiomethyl group having 1 to 4 carbon atoms, more preferably methylthiomethyl; the tri-substituted silyl group such as a tri (lower alkylsilyl) group (eg, trimethylsilyl, tietrylsilyl, tributylsilyl, tert-butyldimethylsilyl, tri-tert-butylsilyl, etc.) or a (lower alkyl) -diarylsilyl group (e.g. , methyldiphenylsilyl, ethyldiphenylsilyl, propyldiphenylsilyl, tert-butyldiphenylsilyl, etc.), more preferably the tri (alkyl of 1 to 4 carbon atoms) silyl group and the (C1-4 alkyl) diphenylsilyl group, more preferably the group tert-butyldimethylsilyl and the tert-butyldiphenylsilyl group; and an acyl group such as an aliphatic, aromatic acyl group or an aliphatic acyl group substituted with an aromatic group, which are carboxylic acid, sulphonic acid or carbamic acid derivatives. Examples of the aliphatic acyl groups include a lower alkanoyl group optionally having one or more suitable substituents such as carboxyl, for example, formyl, acetyl, propionyl, butyryl, isobutyryl, valeryl, isovaleryl, pivaloyl, hexanoyl, carboxyacetyl, carboxypropionyl, carboxybutyryl , carboxyhexanoyl, etc.; a cyclo (lower alkoxy) (lower alkanoyl) group optionally having one or more suitable substituents such as lower alkyl, eg, cyclopropyloxyacetyl, cyclobutyloxypropionyl, cycloheptyloxybutyryl, menthyloxyacetyl, menthyloxypropionyl, menthyloxybutyryl, menthyloxypentanoyl, menthyloxyhexanoyl, etc .; a camphorsulfonyl group; or a lower alkylcarbamoyl group having one or more suitable substituents such as carboxyl or protected carboxyl, for example, carboxy (lower alkyl) carbamoyl (for example, carboxymethylcarbamoyl, carboxyethylcarbamoyl, carboxypropylcarbamoyl, carboxybutylcarbamoyl, carboxypentilcarbamoyl, carboxyhexylcarbamoyl, etc.), the group tri (lower alkyl) silyl (lower alkoxy) carbonyl (lower alkyl) carbamoyl (for example, trimethylsilylmethoxycarbonylethylcarbamoyl, trimethylsilylethoxycarbonylpropylcarbamoyl, triethylsilylethoxycarbonylpropylcarbamoyl, tert-butyldimethylsilylethoxycarbonylpropylcarbamoyl, tri-methylsilylpropoxycarbonylbutylcarbamoyl, etc.) and the like. Examples of the aromatic acyl groups include an aroyl group optionally having one or more suitable substituents such as nitro, for example, benzoyl, toluoyl, xyloyl, naphthoyl, nitrobenzoyl, dinitrobenzoyl, nitronaftoyl, etc .; and an arenesulfonyl group optionally having one or more substituents such as halogen, for example, benzenesulfonyl, toluenesulfonyl, xylene sulfonyl, naphthalenesulfonyl, fluorobenzenesulfonyl, chlorobenzenesulfonyl, bromobenzenesulfonyl, iodobenzenesulfonyl, etc. Examples of the aliphatic acyl groups substituted by an aromatic group include the aryl (lower alkanoyl) group optionally having one or more suitable substituents such as lower alkoxy or trihalo (lower alkyl), for example, phenylacetyl, phenylpropionyl, phenylbutyryl, 2 - trifluoromethyl-2-methoxy-2-phenylacetyl, 2-ethyl-2-trifluoromethyl-2-phenylacetyl, 2-trifluoromethyl-2-propoxy-2-phenylacetyl, etc. More preferably, the acyl groups between the aforementioned acyl groups are the alkanoyl group of 1 to 4 carbon atoms optionally having carboxyl group, cyclo (alkoxy of 5 to 6 carbon atoms) (alkanoyl of 1 to 4 carbon atoms) having two alkyl groups of 1 to 4 carbon atoms in the cycloalkyl moiety, camphorsulfonyl group, carboxy (alkyl of 1 to 4 carbon atoms) carbamoyl, tri (alkyl of 1 to 4 carbon atoms) silyl (alkoxycarbonyl) group from 1 to 4 carbon atoms) (alkylcarbamoyl of 1 to 4 carbon atoms, benzoyl group optionally having one or two nitro groups, benzenesulfonyl group having halogen, or phenyl (alkanoyl of 1 to 4 carbon atoms) having alkoxy from 1 to 4 carbon atoms and the trihalo group (alkyl of 1 to 4 carbon atoms.) Among these, the most preferred are the acetyl, carboxypropionyl, menthyloxyacetyl, camphorsulfonyl, benzoyl, nitrobenzoyl, dinitrobenzoyl, iodobenzene groups. n-sulfonyl and 2-trifluoromethyl-2-methoxy-2-phenylacetyl. Preferable examples of the "saturated or unsaturated 5 or 6 membered heterocyclic ring containing nitrogen, sulfur and / or oxygen atoms" include a pyrrolyl group and a tetrahydrofuryl group. R24 is an optionally substituted heterocyclic ring which may contain one or more heteroatoms, and preferably R24 may be the cyclo group (alkyl of 5 to 7 carbon atoms), and the following may be exemplified: a) a group 3, 4- di-oxo-cyclohexyl; b) a group 3-R20-4-R21-cyclohexyl, in which R20 is hydroxyl, an alkoxy group, an oxo group, or a group -OCH2OCH2CH2OCH3, and r21 is hydroxyl, -OCN, an alkoxy group, a heteroaryloxy may be substituted with suitable substituents, 1- or 2-tetrazolyl, a group -OCH2OCH2CH2OCH3, a protected hydroxyl group, chlorine, bromine, iodine, aminooxalyloxy, an azido group, p-tolyloxythiocarbonyloxy, or R25R6CHCOO- (in which R25 is the optionally protected hydroxyl group, and R26 is hydrogen or methyl, or R20 and R21 combine with one another to form an oxygen atom on an epoxide ring, or c) a cyclopentyl group substituted with methoxymethyl, optionally protected hydroxymethyl, acyloxymethyl (at which acyl portion optionally contains either the dimethylamino group which may be quaternized, or the carboxyl group which may be esterified), one or more amino and / or hydroxyl groups which may be protected, or amino-oxyalyloxymethyl. A preferred example is a 2-formyl-cyclopentyl group. A "heteroaryl which may be substituted with suitable substituents" portion of the "heteroaryloxy which may be substituted with suitable substituents" may be one of those exemplified by R 1 of the compound of formula I of European Patent EP-A-532,088, giving preference to the l-hydroxyethylindol-5-yl group, the description of which is incorporated by reference herein.

The tricyclic compounds (I) and their pharmaceutically acceptable salt for use in the present invention are well known to have excellent immunosuppressive activity, antimicrobial activity and other pharmacological activities and, as such, are of value for the treatment or prevention of rejection reactions by organ or tissue transplantation, graft versus host diseases, autoimmune diseases, and infectious diseases with a method of producing them [European Patent EP-A-0184162, EP-A-0323042, EP-A-423714, EP-A- 427680, EP-A-465426, EP-A-480623, EP-A-532088, EP-A-532089, EP-A-569337, EP-A-626385, WO89 / 05303, O93 / 05058, W096 / 31514 , W091 / 13889, 091/19495, O93 / 04680, O93 / 5059, etc.], the descriptions of which are incorporated by reference herein. Particularly, the compounds that are designed as tacrolimus, FR900520 (ascomycin), FR900523, and FR900525 are products produced by microorganisms of the genus Streptomyces, such as Streptomyces tsukubaensi s No. 9993 (deposited with the National Institute of Bioscience and Human Technology Agency of Industrial Science and Technology (formerly Fermentation Research Institute Industrial Science and Technology Agency), in 1-3, Higashi 1-chome, Tsukuba-shi, Ibaraki, Japan, deposit date October 5, '1984, access number FERM BP-927) or Streptomyces hygroscopicus subsp. yakushimaensi s No. 7238 (deposited with the National Institute of Bioscience and Human Technology, Agency of Industrial Science and Technology (formerly Fermentation Research Institute Industrial Science and Technology Agency), at 1-3, Higashi 1-chome, Tsukuba-shi, Ibaraki, Japan , deposit date January 12, 1985, accession number FERM BP-928) (EP-A-0184162). The tacrolimus of the following chemical formula in particular, is a representative compound.

Chemical Name: 17-allyl-1,4-dihydroxy-12 - [2 - (-hydroxy-3-methoxycyclohexyl) -1-methylvinyl] -23,25-dimethoxy-13,21,21,27-tetramethyl-ll, 28-dioxa-4-azatriciclo [22.3.1. O4.9] octacos-18-en-2, 3,10,16-tetraone. Preferred examples of the tricyclic compounds (I) are those in which each of the adjacent pairs of R3 and R4, and / or R5 and R6 independently form another bond formed between the carbon atoms to which they are attached; each of R8 and R23 are independently a hydrogen atom; R9 is a hydroxyl group; R10 is a methyl group, an ethyl group, a propyl group or an allyl group; X is an oxo group, a state where a hydrogen atom and a hydroxyl group are attached to a carbon atom, a state where two hydrogen atoms are attached to a carbon atom, or to an oxo group; And it's an oxo group; each of R14, R15, R16, R17, R18, R19 and R22 is a methyl group; R24 is a group 3-R20-4-R21-cyclohexyl, in which R20 is the hydroxyl group, an alkoxy group, an oxo group, or a group -OCH2OCH2CH2OCH3, and R21 is hydroxyl, -OCN, an alkoxy group, an heteroaryloxy which can be substituted with suitable substituents, a 1- or 2-tetrazolyl group or a group -OCH2OCH2CH2OCH3, a protected hydroxyl group, chlorine, bromine, iodine, aminooxalyloxy, an azido group, p-tolyloxythiocarbonyloxy, or R25R26CHCOO-, in the which R25 is the optionally protected hydroxyl group or protected amino, and R26 is hydrogen or methyl, or R20 and R21 together form an oxygen atom in an epoxide ring; and n is an integer of 1 or 2. The most preferred tricyclic compounds (I) are, in addition to tacrolimas, the ascomycin derivatives such as halogenated ascomycin (ASM 981) (e.g., 33 -epi-chloro-33-deoxiascomycin) , which is described in European Patent EP-A-427680, Example 66a, 32-0- (l-hydroxyethylindol-5-yl) ascomycin (L-732,531), which is described in European Patent EP-A-532088, 32- (1H-tetrazolyl-1-yl) ascomycin (ABT281) which is described in WO93 / 04680, etc. As the other preferable effects of the macrolide compounds, rapamycin (THE MERCK INDEX (12th edition), No. 8288) and its derivatives can be exemplified. The preferable example of the derivatives is an O-substituted derivative in which the hydroxyl at position 40 of formula A illustrated on page 1 of 095/16691 is replaced with -ORi in which Rx is hydroxyalkyl, hydroalkoxyalkyl, acylaminoalkyl or aminoalkyl; for example, 40-O- (2-hydroxy) ethyl-rapamycin, 40-O- (3-hydroxy) propyl-rapamycin, 40-O- [2- (2-hydroxy) ethoxy] ethyl-rapamycin and 40-O - (2-acetaminoethyl) -rapamycin. These O-substituted derivatives can be produced by the reaction of rapamycin (ot dihydro or deoxo-rapamycin) with an organic radical linked to the leaving group (for example RX where R is an organic radical that is desired as the 0-substituent, such as the alkyl, allyl, or benzyl moiety, and X is a leaving group such as CC13C (NH) O or CF3S03) under the appropriate reaction conditions. The conditions can be acidic or neutral conditions, for example in the presence of an acid such as trifluoromethanesulfonic acid, camphorsulfonic acid, p-toluenesulfonic acid or their respective pyridinium or substituted pyridinium salts when X is CC13C (NH) 0 or in the presence of a base such as pyridine, a substituted pyridine, diisopropylethylamine or pentamethylpiperidine when X is CF3S03. The most preferable is 40-O- (2-hydroxy) ethyl -rapamycin, which is described in O94 / 09010, which is incorporated by reference herein.

The tricyclic compounds (I), and rapamycin and their derivatives, have a similar basic structure, for example the structure of the tricyclic macrolide, and at least one of the similar biological properties for example immunosuppressive activity). The pharmaceutically acceptable salt of the tricyclic compound (I), rapamycin and its derivatives can be a conventional non-toxic and pharmaceutically acceptable salt such as a salt with inorganic or organic bases, specifically, an alkali metal salt such as the sodium salt and the potassium salt, an alkaline earth metal salt such as the calcium salt and the magnesium salt, an ammonium salt and the amine salt such as a triethylamine salt and the N-benzyl-N-methylamine salt. With respect to the pipecolic acid derivatives and the macrolide compounds used in the present invention, it is understood that conformers may exist, and one or more pairs of stereoisomers such as optical isomers, due to one or more asymmetric carbon atoms and geometric isomers due to to one or more double bonds, and such conformers and isomers are also included within the scope of the present invention. In addition, the pipecolic acid derivatives and the macrolide compounds may be in the form I of a solvate, which is included within the scope of the present invention. The solvate preferably includes, for example, a hydrate and an ethanolate. In addition, examples of the pipecolic acid derivatives, which may be used for the purpose of the present invention, include the following: 1) the following Way-124466 compound synthesized by the reaction between 4-phenyl-1, 2, 4-triazolin-3, 5-dione and rapamycin (Ocain et al., Biochemical and Biophysical Research Communications, vol 192, No. 3, 1993); 2) the compound derived from pipecolic acid referred to as RAP-Pa (Carkraborty et al., Chemistry and Biology, March 1995, 2: 157-161); 3) the following compound derived from pipecolic acid (Ikeda et al., J. Am. Chem. Soc. 1994, 116, 4143-4144); 4) ang et al., Bioorganic and Medicinal Chemistry Letters, Vol. 4, No. 9, pp. 1161-1166, 1994, particularly the compounds derived from pipecolic acid described as the compounds 2a-2d; 5) the following pipecolic acid derivative (Birkenshaw et al., Bioorganic and Medicinal Chemistry Letters, Vol. 4, No. 21, pp. 2501-2506, 1994); 6) Holt et al., J. Am. Chem. Soc. 1993, 115, 9925-9938, particularly the compounds derived from pipecolic acid such as compounds 4-14; 7) the compounds derived from pipecolic acid described in Caffer et al., Bioorganic and Medicinal Chemistry Letters, Vol. 4, No. 21, pp. 2507-2510, 1994; 8) compounds derived from pipecolic acid described in Teague et al., Bioorganic and Medicinal Chemistry Letters, Vol. 3, No. 10, pp. 1947-1950, 1993; • i 9) Yamashita et al., Bioorganic and Medicinal Chemistry Letters, Vol. 4, No. 2, pp. 325-328, 1994, particularly the compounds derived from pipecolic acid described as compounds 11, 12 and 19; 10) Holt et al., Bioorganic and Medicinal Chemistry Letters, Vol. 4, No. 2, pp. 315-320, 1994, particularly the compounds derived from pipecolic acid such as compounds 3-21 and 23-24; 11) Holt et al., Bioorganic and Medicinal Chemistry Letters, Vol. 3, No. 10, p. 1977-1980, 1993, particularly the compounds derived from pipecolic acid described as compounds 3-15; 12) Hauske et al., J. Med. Chem. 1992, 35, 4284-4296, particularly the compounds derived from pipecolic acid described as compounds 6, 9-10, 21-24, 26, 28, 31-32 and 52-55; 13) the pipecolic acid derivatives described in Teague et al., Bioorganic and Medicinal Chemistry Letters, Vol. 4, No. 13, pp. 1581-1584, 1994; and 14) Stocks et al., Bioorganic and Medicinal Chemistry Letters, Vol. 4, No. 12, 1457-1460, 1994, particularly the compounds derived from pipecolic acid described as compounds 2, 15-17. The liposomal preparation of the present invention contains essentially the above-described pipecolic acid derivative, trapped within liposomes, and other conditions such as the structure, composition, production method and size of liposomes, and the types of compounds that can be used in combination with liposomes are not specifically limited, unless these conditions adversely affect rapid action of the drug and unless these conditions ensure stable entrapment of the pipecolic acid derivatives within liposomes. Thus, the liposome structure can be a large unilamellar vesicle (LUV), a multilamellar vesicle (MLV) or a small unilamellar vesicle (SUV). Therefore, the size can be in the range of 200 to 1,000 nm for LUV, from 400 to 3,500 nm for MLV and from 20 to 50 nm for SUV in particle diameter. The SUV, which shows low accumulation within a reticuloendothelial system (RES) is preferred. As the liposome that constitutes the liposomal structure, phospholipids and nitrolipids are used. In general, phospholipids are preferred. Examples thereof include natural phospholipids such as egg yolk lecithin (phosphatidylcholine), soy lecithin, lysolecithin, sphingomyelin, phosphatidic acid, phosphatidylserine, phosphatidylglycerol, phosphatidylinositol, phosphatidylethanolamine, diphosphatidylglycerol, cardiolipin, plasmalogen, and so on, or products of hydrogenation obtainable from said phospholipids by conventional technology; and synthetic phospholipids such as dicetyl phosphate, distearoylphosphatidylcholine, dipalmitoylphosphatidylcholine, dipalmitoylphosphatidylethanolamine, dipalmitoylphosphatidylserine, elewanoaroylphosphatidylcholine, elewanoaroylphosphatidylethanolamine, elewanoaroylphosphatidylserine and so on, most preferred include lecithins, and most preferred is egg yolk lecithin. The lipids that include these phospholipids can be used alone, or two or more types of them can be used in combination. In this case, the lipids in which the electronegative group in the phosphatidyl group and the electropositive group in the atomic group (ethanolamine, choline and the like) bonded thereto, are electrically balanced so that the entire molecule is electrically neutral. For example, lecithins, sphingomyelin, phosphatidylethanolamine, distearylphosphatidylcholine and the like are frequently used alone. In contrast, lipids that are electronegative as a whole, in which the atopic group (such as serine, glycerol, inositol or the like) combined with the phosphatidyl group (electronegative group) is electrically neutral, for example, phosphatidylserine, phosphatidylglycerol, phosphatidylinositol and similar or lipids such as phosphatidic acid or dicetyl phosphate, which are electronegative, can be used independently as the lipid in this invention, but it is rather recommended that these be used in combination with the neutral lipid such as those mentioned above. Among these, phosphatidic acid and dicetyl phosphate do not act as the main phospholipid in the formation of the liposome but are known as liposome-forming additives. Considering the stability and handling of the liposomal preparation in the present invention, additives such as excipients and / or stabilizers may be preferably used. Stabilizers preferably used in the present invention include, for example, stearylamine, α-tocopherol, gangliosides, acid glycolipid sulfatides; a type of acid glycolipid and glycolipids that have a sulfuric acid group.

The excipients preferably used in the present invention include, for example, a solid pharmaceutically acceptable saccharide such as monosaccharide (e.g., dextrose, galactose, sorbitol, xylitol, mannitol), disaccharide (e.g., sucrose, lactose, maltose, trehalose), among these, lactose and maltose may be more preferably used, especially maltose may be more preferably used. When the liposome is prepared by the use of cholesterols, a rapid action can hardly be obtained at the same level as that which is going to be required in the present invention and it is likely that cholesterols generally exert undesirable influence on cerebral infarction. Therefore, cholesterols are not used in the present invention. It is also possible to use a-tocopherol with the expectation of a function for an antioxidant. A weight ratio of the pipecolic acid derivative to the lipid for the formation of the liposomal structure is not specifically limited as long as a liposomal preparation which is capable of exerting the desired rapid solubility, can be prepared, but is within the range of from 1: 1 to 1: 500, more preferably from 1: 5 to 1: 100, and more preferably from 1:10 to 1:40. Considering the stability and handling of the liposomal preparation in the present invention, an excipient is preferably used. The amount of the excipient is not specifically limited, but the weight ratio of the pipecolic acid derivative to the stabilizer is within the range of from 1:10 to 1: 1000, more preferably from 1:20 to 1: 500, and most preferably from 1: 100 to 1: 400. To stabilize the liposome, the amount of the stabilizer to be added is not specifically limited, but the weight percentage of the stabilizer to the liposome-forming lipid is preferably within the range of 0.01 to 5%, more preferably 0.05 to 2%, and most preferably from 0.1 to 1%. In view of storage stability, the liposomal preparation of the present invention is preferably provided in the form of an anhydrous product (e.g., a lyophilized product) which is free of the solvent, more preferably in the form of a solid preparation having the feature that the liposomal preparation which includes a pipecolic acid derivative or a pharmaceutically acceptable salt thereof is dispersed in a pharmaceutically acceptable saccharide as an excipient. In use, the liposomal preparation is for example, intravenously injected after being redispersed in a solvent such as distilled water for injection. In the case where cerebral ischemic diseases such as cerebral infarction are treated using the preparation of the present invention, administration of intravenous bolus or administration of rapid infusion is most preferred. The concentration of the pipecolic acid derivative is preferably adjusted within the range of 0.01 to 100 mg / ml, more preferably 0.05 to 50 mg / ml, and most preferably 0.1 to 10 mg / ml, after the redispersion is complete. On the other hand, the dose of the pipecolic acid derivative to be administered by the liposomal preparation of the present invention is appropriately adjusted depending on the type of derivative, the age of the patients, and the conditions of the diseases that will be treated. In the case where the pipecolic acid derivative is a tricyclic compound (I), the liposomal preparation is preferably administered in a daily dose within the range of from about 0.01 to 1000 mg, preferably from 0.1 to 500 mg, and more preferably from 0.5 to 500 mg. 100 mg. A particle size of the liposome is not specifically limited, so long as the size is within the range of a pharmaceutically acceptable size and in such a range as to exert a rapid action. The size of the liposome may preferably be from 1 to 200 nm, more preferably from 10 to 100 nm, and most preferably from 20 to 80 nm. If necessary, a filter can be used when an injection cylinder is filled with a liposomal preparation redispersed in distilled water for injection. In this case, a conventional filter can be used and the preferred example thereof is a filter with the property of low protein binding, free of pyrogens, and stable, having the pore size of 0.2 μm, 0.22 μm, 0.45. μm, 0.8 μm, or 5 μm (Millipore SLGV025LS, SLHVM25LS, SL.AA025LS, SLSV025LS and the like). The liposomal preparation of the present invention can be prepared by the same steps with the following examples.

EXAMPLES The examples of the present invention will be illustrated, but the present invention is not limited to the following examples, and modifications may be made without departing from the purposes described hereinafter and subsequently, and are also included in the technical scope of the present invention. invention Preparative examples of the compounds of the present invention will be described in detail. 1) Preparative Example 1 (EtOH injection method): A solution obtained by dissolving 20 g of purified lecithin from egg yolk and 1 g of tacrolimus in 200 ml of ethanol, is injected and dispersed in a solution obtained separately by dissolving 200 g of lactose in water for injection (1800 ml) with stirring using a homomixer (homomixer). After filtering the dispersion through a 0.2 μm polycarbonate filter, the ethanol in the filtered dispersion is removed by evaporation under reduced pressure and the water for injection is added to the dispersion to make a total volume of 2000 ml. The obtained dispersion is filtered through a polycarbonate filter of 0.2 μm, each 10 ml of the dispersion is filled in a flask, and then lyophilized. The resulting lyophilized liposome preparation is redispersed in 9 ml of water for injection, and a liposomal dispersion containing 0.5 mg / ml tacrolimus and having an average particle diameter of 61.5 nm (measured by a dynamic light scattering method) is obtained. , Model C370 manufactured by NICOMP Company). 2) Preparative Example 2 (Extrusion method): A solution obtained by dissolving 5 g of purified lecithin from egg yolk and 0.25 g of tacrolimus in 100 ml of water, dried under vacuum to form a thin film. The thin film is dispersed in 500 ml of an aqueous 10% maltose solution using a whirlpool mixer. The dispersion is filtered through polycarbonate filters of 400 nm, 200 nm, 100 nm and 50 nm. Each 10 ml of the filtrate is filled into a bottle, and then lyophilized. The resulting lyophilized liposomal preparation is redispersed in 9 ml of water for injection, and a liposomal dispersion containing 0.5 mg / ml of tacrolimus and having an average particle diameter of 110 nm (measured by a dynamic light scattering method) is obtained. , Model C370 manufactured by NICOMP Company). 3) Preparative Example 3 (high pressure emulsion method): A solution is obtained by dissolving 20 g of egg yolk lecithin and 1 g of tacrolimus in 10 ml of ethanol, drying in vacuum to form a thin film . This thin film is dispersed in 2000 ml of an aqueous solution of 10% maltose using a magnetic stirrer. The dispersion is treated using a high pressure emulsification machine manufactured by Nanomizer Company, every 10 ml of the resulting liposomal emulsion is filled into a flask, and then lyophilized. The resulting lyophilized liposomal preparation is redispersed in 9 ml of water for injection, and a liposomal dispersion containing 0.5 mg / ml tacrolimus and having an average particle diameter of 80 nm (measured by a light scattering method; dynamics, Model C370 manufactured by NICOMP, Co.). 4) Preparative Example 4 In the same manner as in Preparative Example 1, a lyophilized liposomal preparation is obtained from the following prescriptions and using a suitable amount of water for injection, and a liposomal dispersion is prepared.

Tacrolimus 3 mg Purified lecithin from egg yolk 100 mg Lactose monohydrate 1000 mg To complete 1103 mg ) Preparative Example 5 In the same manner as in Preparative Example 3, a lyophilized liposomal preparation is obtained from the following prescriptions and using an amount of water for injection, the liposomal preparations 1) and 2) are prepared.

Tacrolimus 3 mg Purified lecithin from egg yolk 100 mg a-tocopherol 0.3 mg Maltose 1000 mg To complete 1103.3 mg Tacrolimus 5 mg > ) Purified lecithin from egg yolk 100 mg a-tocopherol 0.3 mg Maltose 1000 mg To complete 1105.3 mg The test results will be shown later. A normal rat was anesthetized with halotane. A cannula (PE50) was inserted into the femoral vein for the administration of a drug. The rat was held in a fixed cage. After waiting, the drug obtained in Preparative Example 3 was administered in about 30 seconds. Blood samples were collected from a rat abdominal aorta under halothane anesthesia and cooled with ice, and then whole blood was collected (amount required for approximately 1 cubic centimeter). A blood collection, heparin in the amount of 20 μl (approximately 1000 u / ml) based on 1 ml of whole blood was added. The blood was stored with freezing until the concentration was measured. With respect to brain tissue, the brain was removed after decapitation and the blood adhered to the periphery of the brain was washed and dried using a filter paper, and then the wet weight of the brain was weighed. The brain was placed in a centrifuge tube and stored with freezing until the measurement was made. The cerebral cortex was homogenized with 9 times the amount of distilled water as the wet weight of the brain on the day of the presentation of the specimen. The drug concentration of the sample was determined using an enzyme immunoassay described in Unexamined Patent Publication (Kokai) No. 1-92659. As it is apparent from the results, sufficient concentration in the brain or that in the blood has been reached within a short period and a sufficient rapid action is exerted. The test results of A, the concentration of a liposomal preparation containing tacrolimus in the brain and B that in blood, are shown in Figure 1.

BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 is a graph showing A the concentration of a liposomal preparation containing tacrolimus in the brain and B that in blood.

BEST MODALITY TO CARRY OUT THE INVENTION The present invention is directed to a liposomal preparation comprising, as an active ingredient, a pipecolic acid derivative represented by. a macrolide compound of the following general formula (I) or a pharmaceutically acceptable salt thereof trapped in a liposome. A liposomal preparation containing, as an active ingredient, 17-allyl-1,4-dihydroxy-12- [2- (4-hydroxy-3-methoxycyclohexyl) -1-methylvinyl] -23,25-dimethoxy-13, 19 , 21, 27-tetramethyl-ll, 28-dioxa-4-azatricyclo [22.3.1. O 4,9] octacos718-en-2, 3,10, 16-tetraone of the formula (I) above or a pharmaceutically acceptable salt thereof, is particularly preferred. With a preferred constitution, lecithin is primarily used as a liposome-forming lipid and the preparation does not contain cholesterol as a stabilizer. Especially preferred is one in the form of a solid preparation in which a liposomal preparation is dispersed in a pharmaceutically acceptable saccharide, the liposomal preparation is composed of a liposome containing lecithin as a main component and free of cholesterol, and an acid derivative pipecolic or a pharmaceutically acceptable salt thereof is entrapped in the liposome.

POSSIBILITY OF INDUSTRIAL APPLICATION According to the present invention, since the liposome can be easily disintegrated compared to a conventional liposome preparation containing cholesterol, it is possible to expect an excellent fast action by bolus administration. Furthermore, the preparation of the present invention does not contain a surfactant, so that it becomes possible to obtain an excellent effect without exerting any influence on the circulatory organs. In consecuense, the liposomal preparation of the present invention is particularly useful for the treatment and prevention of diseases where a rapid action of drug efficacy is expected, for example, cerebral ischemic diseases (for example head injury, brain hemorrhage (eg. example subarachnoid hemorrhage, intracerebral hemorrhage), cerebral infarction, cerebral thrombosis, cerebral embolism, cardiac arrest, stroke, transient ischemic attack (TIA), hypertensive encephalopathy). Based on the pharmacological effect of a pipecolic acid derivative as an active ingredient, particularly a tricyclic compound (I), the liposomal preparation of the present invention is useful for the treatment and prevention of the following diseases and conditions: rejection reactions by transplantation of organs or tissues such as heart, kidney, liver, bone marrow, skin, cornea, lung, pancreas, small intestine, lower extremities, muscle, nerve, intravertebral disc, trachea, myoblasts, cartilage, etc.; graft versus host reactions after bone marrow transplantation; autoimmune diseases such as rheumatoid arthritis, systemic lupus erythematosus, Hashimoto's thyroiditis, multiple sclerosis, myasthenia gravis, type I diabetes, etc .; infections caused by pathogenic microorganisms (for example, Aspergi llus fumiga tus, Fusarium oxysporum, Tri chophyton as teroides, etc.); inflammatory or hyperproliferative skin diseases or cutaneous manifestations of immunologically mediated diseases (eg, psoriasis, atopic dermatitis, contact dermatitis, eczematoid dermatitis, seborrheic dermatitis, lichen planus, pemphigus, bullous pemphigoid, epidermolysis bullosa, urticaria, angioedema, vasculitides, erythema, dermal eosinophilia, lupus erythematosus, acne and alopecia areata); autoimmune diseases of the eye (eg, keratoconjunctivitis, vernal conjunctivitis, uveitis associated with Behcet's disease, keratitis, herpetic keratitis, conical keratitis, corneal epithelial dystrophy, keratoleukoma, ocular pemphigus, Mooren's ulcer, scleritis, Graves' ophthalmopathy, of Vogt-Koyanagi-Harada, dry keratoconjunctivitis (dry eye), flicténule, iridocyclitis, sarcoidosis, endocrine ophthalmopathy, etc.); reversible obstructive diseases of the respiratory tract [asthma (eg, bronchial asthma, allergic asthma, intrinsic asthma, extrinsic asthma, and powder asthma), particularly chronic or inveterate asthma (eg delayed asthma and hyper-responsiveness of the respiratory tract) bronchitis, etc.]; mucosal or vascular inflammations (eg gastric ulcer, ischemic or thrombotic vascular damage, ischemic bowel diseases, enteritis, necrotizing esterocolitis, intestinal damage associated with thermal burns, diseases mediated by leukotriene B4); intestinal inflammations / allergies (eg abdominal diseases, proctitis, eosinophilic gastroenteritis, mastocytosis, Crohn's disease and ulcerative colitis); allergic diseases related to food with symptomatic manifestation of the gastrointestinal tract (for example migraine, rhinitis and eczema); kidney diseases (for example intestinal nephritis, Goodpasture syndrome, hemolytic uremic syndrome, diabetic nephropathy), and nephrotic syndrome (for example glomerulonephritis); nerve diseases (eg multiple myositis, Guillain-Barre syndrome, Meniere's disease, multiple neuritis, solitary neuritis, cerebral infarction, Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis (ALS and radiculopathy), endocrine diseases (eg hyperthyroidism) , and Basedow's disease), blood diseases (eg, pure red cell aplasia, aplastic anemias, hypoplastic anemia, idiopathic thrombocytopenic purpura, autoimmune hemolytic anemia, agranulocytosis, pernicious anemia, megaloblastic anemia, and aneurysplasia), bone diseases (eg, osteoporosis) ), respiratory diseases (eg sarcoidosis, pulmonary fibrosis, and idiopathic interstitial pneumonia), skin diseases (eg dermatoraiositis, leucoderma vulgaris, ichthyosis vulgaris, photosensitivity, and cutaneous T-cell lymphoma), - circulatory diseases (eg arteriosclerosis) , atheroscler osis, aortitis syndrome, polyarteritis nodosa, and myocardosis); collagen diseases (eg, scleroderma, Wegener's granuloma, and Sj ogren's syndrome); adiposis; eosinophilic fasciitis periodontal diseases (eg damage to the gingiva, periodontium, alveolar bone or dentis of the bone substance); male pattern alopecia, senile alopecia; muscular dystrophy; pyoderma and Sezary syndrome; diseases associated with chromosomal abnormalities (for example Down syndrome); Addison's disease; active oxygen-mediated diseases [eg organ damage (eg, ischemic disorders of the circulation of organs (eg, heart, liver, kidney, digestive tract, etc.) associated with preservation, transplantation, or ischemic diseases (eg. example, thrombosis, cardiac infarction, etc.)); intestinal diseases (e.g. endotoxin shock, pseudomembranous colitis, and drug-induced or radiation-induced colitis); kidney diseases (eg, ischemic acute renal failure, and chronic renal failure); lung diseases (eg, toxicosis caused by oxygen or pulmonary drugs (eg, paracort, bleomycin, etc.), lung cancer, and pulmonary emphysema); eye diseases (eg, cataract, iron storage disease (bulbi siderosis), retinitis pigmentosa, senile plaques, vitreous scarring, corneal alkali burn); dermatitis (e.g., erythema multiforme, bullous dermatitis due to linear immunoglobulin A, cement dermatitis); and other diseases (eg, gingivitis, periodontitis, sepsis, pancreatitis, and diseases caused by environmental pollution (eg, air pollution), aging, carcinogens, carcinoma metastases, and hypobaropathy)]; diseases caused by release of histamine or leukotriene C4; coronary artery restenosis after angioplasty and prevention of post-surgical adhesions; autoimmune diseases and inflammatory conditions (eg, primary mucosal edema, autoimmune atrophic gastritis, premature menopause, male sterility, juvenile diabetes mellitus, pemphigus vulgaris, pemphigoid, sympathetic ophthalmitis, lens induced uveitis, idiopathic leukopenia, chronic active hepatitis, cirrhosis idiopathic, discoid lupus erythematosus, autoimmune orchitis, arthritis (eg deforming arthritis), or polychondritis); infection by Virus (HIV), AIDS; allergic conjunctivitis; and hypertrophic and keloid scar due to trauma, burn, or surgery. In addition, macrolide compounds such as the tricyclic compound (I) have liver regeneration activity and / or hepatocyte hypertrophy and hyperplasia stimulating activities. Therefore, the pharmaceutical liposomal preparation of the present invention is useful for increasing the effect of therapy and / or prophylaxis of liver diseases [e.g., immunogenic diseases (e.g. chronic autoimmune liver diseases such as autoimmune liver diseases, biliary cirrhosis. primary or sclerosing cholangitis), partial liver resection, acute hepatic necrosis (eg, necrosis caused by toxins, viral hepatitis, shock, or anoxia), hepatitis B, hepatitis A A non B, hepatocirrosis, and liver failure (eg, fulminant hepatitis, late-onset hepatitis and "acute on chronic" liver failure (acute liver failure on chronic liver diseases))]. And in addition, the present liposomal composition is also useful to increase the effect of the prevention and / or treatment of various diseases, due to the useful pharmacological activity of said tricyclic macrolides, such as the increase of the activity of the chemotherapeutic effect, infection activity by cytomegalovirus, anti-inflammatory activity, inhibition of activity against peptidyl-prolyl isomerase or rotamase, antimalarial activity, antitumor activity, and the like.

Claims (11)

1. A liposomal preparation containing a pipecolic acid derivative, comprising a pipecolic acid derivative or a pharmaceutically acceptable salt thereof trapped within liposomes.

2. The liposomal preparation according to claim 1, wherein the lecithin is mainly used as a lipid for liposome formation.

3. The liposomal preparation according to claim 2, wherein the liposome does not substantially contain cholesterol.

4. The liposomal preparation according to claim 1, wherein the pipecolic acid derivative is a macrolide compound of the following general formula (I) or a pharmaceutically acceptable salt thereof: N wherein each of the adjacent pairs of R1 and R2, R and R4, and R5 and R6 independently: a) are two adjacent hydrogen atoms; or b) they can form another bond formed between the carbon atoms to which they are attached; and R2 may also be an alkyl group; R7 is a hydrogen atom, a hydroxyl group, a protected hydroxyl group, or an alkoxy group, or an oxo group together with R1; R8 and R9 are independently a hydrogen atom or a hydroxyl group; R10 is a hydrogen atom, an alkyl group, an alkyl group substituted with one or more hydroxyl groups, an alkenyl group, an alkenyl group substituted with one or more hydroxyl groups, or an alkyl group substituted with an oxo group; X is an oxo group, a state where a hydrogen atom and a hydroxyl group are attached to a carbon atom, a state where two hydrogen atoms are attached to a carbon atom, or a group represented by the formula -CH20-; And it is an oxo group, a state where a hydrogen atom and a hydroxyl group are attached to a carbon atom, a state where two hydrogen atoms are attached to a carbon atom, or a group represented by the formula N-NR R? 2 Q N_OR13; R11 and R12 are independently a hydrogen atom, an alkyl group, an aryl group, or a tosyl group; R13, R14, R15, R16, R17, R18, R19, R22 and R23 are independently a hydrogen atom or an alkyl group; R24 is an optionally substituted heterocyclic ring which may contain one or. more heteroatoms; and n represents an integer of 1 or 2. with the proviso that, in addition to the above definitions, Y, R 10 and R 23, together with the carbon atoms to which they are attached, can represent a saturated or unsaturated heterocyclic ring of 5. or of 6 members containing nitrogen, sulfur and / or oxygen atoms, and the heterocyclic ring may be substituted with one or more groups selected from the group consisting of an alkyl group, a hydroxyl group, an alkyloxy group, a benzyl group, a group of the formula -CH2Se (C6H5), and an alkyl substituted with one or more hydroxyl groups.

5. The liposomal preparation according to claim 4, wherein the macrolide compound (I) is 17-allyl-1,4-dihydroxy-12 - [2- (4-hydroxy-3-methoxycyclohexyl) -1-methylvinyl] -23 , 25-dimethoxy-13, 19,21,27-tetramethyl-II, 28-dioxa-4-azatricyclo [22.3.1.0 '9] octacos-18-en-2,3,10,16-tetraone or its hydride.

6. A solid preparation in which the liposomal preparation as defined according to any of claims 1 to 5, is dispersed in a pharmaceutically acceptable saccharide.

7. A solid preparation comprising a liposomal preparation dispersed in a pharmaceutically acceptable saccharide, wherein the liposomal preparation is composed of a liposome containing lecithin as a major component and free of cholesterol and a pipecolic acid derivative or a pharmaceutically acceptable salt thereof is trapped in the liposome.

8. The liposomal preparation according to claim 7, wherein a mass ratio between a pipecolic acid derivative or a pharmaceutically acceptable salt thereof, the lecithin, and a pharmaceutically acceptable saccharide of the solid is 1: 1-500: 10- 1000

9. The solid preparation according to claim 7 or 8, wherein the saccharide is lactose or maltose.

10. The liposomal preparation according to claim 6, which is a lyophilized product.

11. The liposomal preparation according to claim 1, which is dispersed in distilled water for injection.