Sustained release preparations

Description of the invention

Field of the Invention

The present invention relates to an extremely slow release formulation containing a macrolide-tacrolimus compound or hydrate thereof for use in the medical field.

State of the art

An oral formulation of one of the macrolide compounds, namely tacrolimus with useful immunosuppressive activity, has been prepared as solid dispersion compositions which exhibit fast release characteristics by the use of polymers such as hydroxypropyl methylcellulose and a disintegrant (see e.g. EP 0 240 773). Due to the presence of the disintegrant, it is a fast release formulation. It has been highly appreciated in clinical use due to its high absorbability. In clinical practice, alternatively, the emergence of an oral tacrolimus formulation with a sufficiently long duration of action and excellent oral absorption was expected.

However, according to the state of the art for the skilled person, the absorbability of a pharmaceutically active agent administered orally as a slow release preparation is generally reduced and / or a non-negligible change in absorbability is observed. The authors of the present invention have conducted a great deal of research. Consequently, the inventors have developed formulations with a slow release of macrolide compounds-tacrolimus or its hydrate, characterized in that this macrolide compound is orally absorbed excellently and / or the modification of its absorbability is blocked.

Disclosure of the Invention

The present invention relates to a formulation for the slow release of a macrolide-tacrolimus compound or a hydrate thereof, wherein dissolution of the macrolide compound is carried out by slow release.

The invention relates to a slow-release formulation comprising a solid dispersion composition, characterized in that the solid dispersion composition comprises tacrolimus or a hydrate thereof, in a mixture containing a water-soluble polymer, a water-insoluble polymer and auxiliaries, the time T63 , 2% necessary to dissolve 63.2% of the maximum amount of tacrolimus or its hydrate is between 0.7 and 15 hours, measured in accordance with the Japanese Pharmacopoeia, 13th edition, dissolution test no. 2, paddle stirrer test, 50 rpm, using a test solution which is a 0.005% aqueous solution of hydroxypropyl cellulose at pH 4.5.

The slow release formulation is preferably characterized in that the solid dispersion composition has a particle size equal to or less than 250 µm.

Preferably, the slow-release formulation has a weight ratio of water-soluble polymer to tacrolimus of 0.2-0.4: 1 and the weight ratio of water-insoluble polymer to tacrolimus is 0.1-5: 1, preferably a weight ratio of water-insoluble polymer to tacrolimus. polymer to tacrolimus is 0.1-1: 1.

Preferably, the water-insoluble polymer in the slow release formulation is ethyl cellulose.

The formulation preferably comprises a water-soluble polymer which is hydroxypropyl methylcellulose.

Preferably the excipient in the slow release formulation is lactose.

Preferably the formulation has a weight ratio of lactose to tacrolimus of 2, 3 or 5: 1 in the formulation.

The formulation preferably comprises a solid dispersion composition substantially free of disintegrants.

The sustained release formulation preferably comprises tacrolimus or a hydrate thereof, which is present in an amorphous state.

Preferably in the formulation, the weight ratio of water-soluble polymer to tacrolimus is 0.2-0.4: 1, and the weight ratio of water-insoluble polymer to tacrolimus is 0.1-5: 1, especially the weight ratio of water-insoluble polymer to tacrolimus is 0.1-1: 1.

PL 193 244 B1

Preferably the water-insoluble polymer in the formulation is ethyl cellulose.

Lactose is preferably used as the excipient in the formulation.

The slow release formulation preferably comprises tacrolimus or its hydrate in amorphous state in a mixture with ethyl cellulose and hydroxypropyl methyl cellulose, preferably the weight ratio of ethyl cellulose to tacrolimus is 0.3: 1 and the weight ratio of hydroxypropyl methyl cellulose to tacrolimus is 0.3: 1.

In a slow release formulation, the excipient is preferably lactose.

The formulation preferably has a weight ratio of lactose to tacrolimus of 2: 1.

In the formulation, preferably the solid dispersion composition has a particle size equal to or less than 212 µm.

In the formulation, preferably the water-insoluble polymer is ethyl cellulose and the water-soluble polymer is hydroxypropyl methyl cellulose.

The slow release preparation is preferably in the form of a powder, granule, tablet or capsule.

It is also an object of the invention to provide a solid dispersion composition of said macrolide compound usable in a slow release formulation as mentioned above, wherein the macrolide compound is present in an amorphous state in the solid support.

It is a further object of the present invention to provide a fine powder of said macrolide compound having a particle diameter distribution in the range of 0.1 ~ 50 Pm and / or a mean particle diameter in the range 0.2 ~ 20 Pm for use in the above-mentioned slow release formulation.

The T63.2% value determined by the dissolution test in accordance with the present invention can be estimated from the release curve constructed by plotting the test data on graph paper. However, the drug release profile can in general be analyzed by fitting the dissolution test data to the release model, and this method can also be used to calculate the said T-value of 63.2%. The fit model that can be used includes a linear or first-order model, a zero-order model, a cubic root model, etc. as described in Yamaoka, K. & Yagahara, Y .: Introduction to Pharmacokinetics with a Microcomputer, Nankodo, pp. 138, but the model by which all kinds of release patterns can best be expressed is the famous Weibull function which is described in the above book and in: LJ Leeson & JT Carstensen (ed.): Pharmaceutical Product Release (American Pharmaceutical Society) (Chizin Shokan), pp. 192-195.

The Weibull function is a function such that the degree of dissolution (%) with time (T) can be expressed by the following equation:

Dissolution rate (%) = Dmax x {1-exp [- ((T-Ti)ⁿ) / m]} where Dmax is the maximum degree of dissolution at infinite time, m is the scale parameter representing the dissolution rate, n is the shape parameter representing the shape of the dissolution curve, Ti is the position parameter representing the delay time to the onset of dissolution, and the dissolution characteristics of the pharmaceutical product can be expressed by using a combination of these parameters.

To fit the dissolution test data to the Weibull function and calculate the corresponding parameters, the nonlinear least squares method is used as described in Yamaoka, K. & Yagahara, Y. Introduction to Pharmacokinetics with a Microcomputer., Nankodo, p. 40, mentioned above.

In particular, the parameters are determined at a time point where the sum of the squared differences between the values ​​calculated by the above equation and the values ​​measured at each time point is minimal and the dissolution curve calculated by the above equation using these parameters is the curve that most faithfully reproduces the measured values.

The meaning of each parameter of the Weibull function will now be explained.

Dmax- (maximum degree of dissolution) is the maximum degree of dissolution in infinite time as mentioned above and in general the Dmax value is preferably as close as possible to 100% (%).

m (scale parameter) is a parameter representing the dissolution rate of a pharmaceutical product, and the smaller the m value, the faster the dissolution rate, and similarly, the larger the m value, the slower the dissolution rate.

n (shape parameter) is a parameter representing the shape of the dissolution curve. When the value of n is 1, the Weibull function can be written as the degree of dissolution (%) Dmax x {1-exp [- (T-Ti) / m]), and since this is equivalent to the first order kinetics, the dissolution curve is linear. When

When the value of n is less than 1, the dissolution curve plateaued. When the value of n is greater than 1, the sigmoidal dissolution curve predominates.

Ti (position parameter) is a parameter representing the lag time until the start of dissolution.

A slow release formulation comprising a macrolide-tacrolimus compound or a hydrate thereof according to the present invention can also be characterized by the aforementioned Weibull function. Thus, the subject slow release formulation can be made by setting

Dmax (maximum degree of dissolution) at 80% or more, preferably 90% or more, especially

95% or more m (scale parameter) with 0.7-20, preferably 1-12, especially 1.5-8, n (shape parameter) with 0.2-5, preferably 0.3-3, especially 0.5-1.5, and Ti (position parameter) at 0-12, preferably 0-8, especially 0-4.

The value determined by substituting the values ​​of the parameters min from the above Weibull function to the term m^{1 / n} is the time during which 63.2% of the maximum amount of dissolved active ingredient is released from the formulation (T63.2%). Ie. T63.2% (hr) = m^{1 / n}. The release characteristics of the slow release formulation of the present invention can be assessed from the dissolution test method 2 (paddle method, 50 rpm) of JP XlII using a test solution which is 0.005% aqueous hydroxypropyl cellulose adjusted to pH 4.5 . In a slow release formulation comprising a macrolide compound of the present invention, the time (T63.2%) after which 63.2% of the maximum amount of macrolide compound dissolved from the formulation is 0.7-15 hours. In the past, although a quick release formulation containing the macrolide compound has already been prepared, no slow release formulations with a T63.2% of 0.7-15 hr which would be useful in clinical practice have never been prepared. This is the first time this has been done in the present invention. If the T63.2% value is less than 0.7 hours, the oral efficacy of the macrolide compound will not be slow enough. When the formulation has a T63.2% value over 15 hours, the release of the active ingredient will be so delayed that the active ingredient will be eliminated from the body before an effective blood concentration is reached, and thus it will be unsuitable for a formulation according to the present invention. When T63.2% is 1.0-12 hours, a more favorable slow release can be achieved. More preferably, the T63.2% is 1.3-8.2 hours, and a slow release formulation with a T63.2% value of 2-5 hours is especially preferred.

The term "macrolide compound" is the generic name of compounds with 12 or more members that belong to large ring lactones. Numerous macrolide compounds produced by microorganisms of the genus Streptomyces such as rapamycin, tacrolimus (FK506), and ascomycin, and analogs and derivatives thereof, are encompassed by the term macrolide compound.

As a specific example of a macrolide compound, there can be provided a tricyclic compound of the following formula (I):

<img file="PL193244B1_D0001.tif" />

(where each of the adjacent pairs of R.¹ and r², R³ and r⁴and R.⁵ and r⁶ independently

(A) is two adjacent hydrogen atoms, but R² it may also be alkyl or (b) may form another bond formed between the carbon atoms to which they are attached;

R⁷ is hydrogen, hydroxy, protected hydroxy or alkoxy, or oxo together with R¹;

R⁸ and r⁹ are independently hydrogen or hydroxy;

R¹⁰ is hydrogen, alkyl, alkyl substituted with one or more hydroxy groups, alkenyl, alkenyl substituted with one or more hydroxy groups, or alkyl substituted with oxo;

X is oxo, (hydrogen and hydroxy), (hydrogen and hydrogen), or a group of the formula -CH2O-;

Y is oxo, (hydrogen and hydroxy), (hydrogen and hydrogen), or e of formula N-NR¹¹R¹² or N-OR¹³

R¹¹ and r¹² are independently hydrogen, alkyl, aryl or tosyl;

R¹³, R¹⁴, R¹⁵, R¹⁶, R¹⁷, R¹⁸, R¹⁹, R²² and r²³ are independently hydrogen or alkyl;

R²⁴ is an optionally substituted ring system that may contain one or more heteroatoms; n is an integer of 1 or 2; and in addition to the above terms, Y, R¹⁰ and r²³, together with the carbon atoms to which they are attached, may be a saturated or unsaturated 5- or 6-membered heterocyclic ring containing nitrogen, sulfur and / or oxygen optionally substituted with one or more groups selected from the group consisting of alkyl, hydroxy, alkoxy, benzyl, a group of the formula -CH2Se (C6H5), and alkyl substituted with one or more hydroxy groups.

The preferred R.²⁴ may be cyclo (C5-7) alkyl, and the following groups can be mentioned as examples:

(a) 3,4-di-oxo-cyclohexyl;

(b) 3-R²⁰-4-R²¹-cyclohexyl, wherein R²⁰ is hydroxy, alkoxy, oxo, or -OCH2OCH2CH2OCH3, and

R²¹ is hydroxy, -OCN, alkoxy, heteroaryloxy which may be substituted with suitable substituents, a -OCH2OCH2CH2OCH3 group, a protected hydroxy group, chlorine, bromine, iodine, amino oxalyloxy, azide, p-tolyloxythiocarbonyloxy, or R²⁵R²⁶CHCOO- in which

R²⁵ is an optionally protected hydroxy group or a protected amino group, and

R²⁶ is hydrogen or methyl, or

R²⁰ and r²¹ together they form an oxygen atom in the epoxy ring; or (c) cyclopentyl substituted with methoxymethyl, optionally protected hydroxymethyl, acyloxymethyl (in which the acyl moiety optionally contains either a dimethylamino group which may be quaternized or a carboxy group which may be esterified), one or more amino and / or hydroxy groups. which may be protected, or aminoxalyloxymethyl. A preferred example is a 2-formyl-cyclopentyl group.

The terms used in the above general formula (I) and specific preferred examples thereof are now explained and detailed below.

The term "lower" means, unless otherwise stated, a group having from 1 to 6 carbon atoms.

Preferred examples of the "alkyl groups" and the alkyl moiety of "alkoxy groups" include a straight or branched aliphatic hydrocarbon chain, eg, lower alkyl such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, neopentyl and hexyl.

Preferable examples of "alkenyl groups" include a straight or branched aliphatic hydrocarbon chain with one double bond, eg, lower alkenyl such as vinyl, propenyl (eg, allyl), butenyl, methylpropenyl, pentenyl and hexenyl.

Preferable examples of "aryl groups" include phenyl, tolyl, xylyl, cumene, mesityl and naphthyl.

Preferred protecting groups on "protected hydroxyl" and protected amino groups include 1- (lower alkylthio) - (lower) alkyl, such as lower alkylthiomethyl (e.g., methylthiomethyl, ethylthiomethyl, propylthiomethyl, isopropylthiomethyl, butylthiomethyl, isobutylthiomethyl, etc.), particularly preferably a C1-C4 alkylthiomethyl group, especially a methylthiomethyl group; a trisubstituted silyl group such as tri (lower) alkylsilyl (e.g., trimethylsilyl, triethylsilyl, tributylsilyl, tert-butyldimethylsilyl, tri-tert-butyldimethylsilyl, etc.) or lower alkyldiarylsilyl (e.g., methyldiphenylsilyl, ethenyldiphenyl, etc.) ), in particular a tri (C1-C4) alkylsilyl group and a C1-C4alkyldiphenylsilyl group, in particular a tert-butyldimethylsilyl group

PL 193 244 B1 and tert-butyl-diphenylsilyl; and an acyl group such as an aliphatic, aromatic acyl group, or an aliphatic acyl group substituted with an aromatic group that is derived from a carboxylic acid, sulfonic acid, or carbamic acid.

Examples of aliphatic acyl groups include: lower alkanoyl optionally with one or more suitable substituents such as carboxy, e.g. formyl, acetyl, propionyl, butyryl, isobutyryl, valeryl, isovaleryl, pivaloyl, hexanoyl, carboxyacetyl, carboxypropionyl, carboxybutyryl, carboxyhexanoyl , etc. cyclo (lower) alkoxy (lower) alkanoyl optionally containing one or more suitable substituents such as lower alkyl, e.g. cyclopropyloxyacetyl, cyclobutyloxypropionyl, cycloheptyloxybutyryl, menthyloxyacetyl, menthyloxypropionyl, menthyloxybutyryl, menthyloxypentanoyl, menthyloxyhexanoyl, etc., a camphorsulfonyl group or a lower alkylcarbamoyl group containing one or more suitable carboxylic or protected substituents such as e.g. carboxy (lower) alkylcarbamoyl (e.g., carboxymethylcarbamoyl, carboxyethylcarbamoyl, carboxypropylcarbamoyl, carboxybutylcarbamoyl, carboxypentylcarbamoyl, carboxyhexylcarbamoyl, etc., tri- (lower) alkylsilyl (lower) alkylcarbamoyl (lower) alkoxycarbamoyl, e.g. trimethylsilylmethoxycarbonylethylcarbamoyl, trimethylsilylethoxycarbonylpropylcarbamoyl, triethylsilylethoxycarbonylpropylcarbamoyl, tert-butyl-dimethylsilylethoxycarbonylpropylcarbamoyl, trimethylsilyl-propoxycarbonyl, etc.

Examples of aromatic acyl groups include an aroyl group optionally containing one or more suitable substituents such as nitro, e.g., benzoyl, tolyl, xyloyl, naphthoyl, nitrobenzoyl, dinitrobenzoyl, nitronaphthoyl and the like; and an arenesulfonyl group, optionally containing one or more suitable substituents such as halogen, e.g. benzenesulfonyl, toluenesulfonyl, xylenesulfonyl, naphthalenesulfonyl, fluorobenzenesulfonyl, chlorobenzenesulfonyl, bromobenzenesulfonyl, iodobenzenesulfonyl, etc.

Examples of aliphatic acyl groups substituted with an aromatic group include ar (lower) alkanoyl optionally containing one or more suitable substituents such as lower alkoxy or trihalo (lower) alkyl, e.g. phenylacetyl, phenylpropionyl, phenylbutyryl, 2-trifluoromethyl-2-methoxy -2-phenylacetyl, 2-ethyl-2-trifluoromethyl-2-phenylacetyl, 2-trifluoromethyl-2-propoxy-2-phenylacetyl etc.

Particularly preferred acyl groups from the abovementioned acyl groups: are C1-C4 alkanoyl, optionally carboxy-containing, cyclo (C5-C6) alkoxy (C1-C4) alkanoyl, containing two (C1-C4) alkyls on the cycloalkyl moiety, camphorsulfonyl, carboxy- (C1-C4) alkylcarbamoyl, tri (C1-C4) alkylsilyl (C1-C4) -alkoxycarbonyl (C1-C4) -alkylcarbamoyl, benzoyl group optionally containing one or two nitro groups, halogen-containing benzenesulfonyl group or phenyl (C1-C4) -alkanoyl containing C1-C4 alkoxy and trihalo (C1-C4) alkyl.

Of these, acetyl, carboxypropionyl, menthyloxyacetyl, camphorsulfonyl, benzoyl, nitrobenzoyl, dinitrobenzoyl, iodobenzenesulfonyl and 2-trifluoromethyl-2-methoxy-2-phenylacetyl are particularly preferred.

Preferred examples of the "5- or 6-membered nitrogen, sulfur and / or oxygen containing heterocyclic ring" include a pyrrolyl group and a tetrahydrofuryl group.

The "heteroaryl which may be substituted with suitable substituents" of the "heteroaryloxy which may be substituted with suitable substituents" may be as exemplified for R¹ in a compound of the formula described in EP-A-532,088, preferably 1-hydroxyethylindol-5-yl, the description of which is hereby incorporated by reference into the present application.

The tricyclic compounds (I) and their pharmaceutically acceptable salts for use in the present invention are well known to have excellent immunosuppressive activity, antimicrobial activity, and other pharmacological activities, and as such are valuable in treating or preventing rejection reactions in organ or tissue transplantation, disease graft versus host reaction, autoimmune diseases, and infectious diseases [EP-A0184162, EP-A-0323042, EP-A-423714, EP-A-427680, EP-A-465926, EP-A-480623, EP-A-532088, EP-A-532089, EP-A-569337, EP-A-626385, W089 / 05303, WG93 / 05058, W096 / 31514, W091 / 13889, W091 / 19495, W093 / 5059, etc.).

In particular, the compounds that have been designated FR900506 (= FK506), FR900520 (ascomycin), FR900523, and FR900525 are products produced by microorganisms from a Streptomyces strain such as Streptomyces tsukubaensis No. 9993 [filed with the National Institute of Bioscience and Human Technology Agency of Industrial Science and Technology (formerly Fermentation Research Institute Agency of Industrial Science and Technology), at 1-3, Higashi 1-chome, Tsukubashi, Ibaraki, Japan, deposit date October 5, 1984, accession number FERM BP-927] or Streptomyces hyPL 193 244 B1 groscopicus subsp. yakushimaensis No. 7238 [deposited at the National Institute of Bioscience and

Human Technology Agency of Industrial Science and Technology (formerly Fermentation Research

Institute Agency of Industrial Science and Technology), at 1-3, Higashi 1-chome, Tsukuba-shi, Ibaraki, Japan filed January 12, 1985, accession number FERM BP-928] [EP-A-0184162]. FK506 (generic name: tacrolimus) with the following chemical formula is in particular a representative compound:

<img file="PL193244B1_D0002.tif" />

chemical name: 17-allyl-1,14-dihydroxy-12- [2- (4-hydroxy-3-methoxycyclohexyl) -1-methylvinyl] -23,25-dimethoxy-13,19,21,27-tetramethyl-11 , 28-dioxa-4-azathi-cyclo [22.3.1.0^4,9] octacos-18-ene-2,3,10,16-tetraon.

Preferred examples of the tricyclic compounds (I) are those in which each of the adjacent pairs of R³ and r⁴ or R⁵ and r⁶ independently form another bond formed between the carbon atoms to which they are attached;

each of the R.⁸ and r²³ is independently a hydrogen atom;

R⁹ is hydroxy;

R¹⁰ is methyl, ethyl, propyl, or allyl;

X is (hydrogen and hydrogen) or oxo;

Y is oxo;

each of the R.¹⁴, R¹⁵, R¹⁶, R¹⁷, R¹⁸, R¹⁹, and R.²² is methyl;

R²⁴ is 3-R²⁰-4-R²¹-cyclohexyl, wherein R²⁰ is hydroxy, alkoxy, oxo, or -OCH2OCH2CH2OCH3, and

R²¹ is hydroxy, -OCN, alkoxy, heteroaryloxy, which may be substituted with suitable substituents, -OCH2OCH2CH2OCH3, protected hydroxy, chlorine, bromine, iodine, amino oxalyloxy, azido, p-tolyloxythiocarbonyloxy, or R²⁵R²⁶CHCOO-, in which R.²⁵ is an optionally protected hydroxy group or a protected amino group, and

R²⁶ is hydrogen or methyl, or

21

R²⁰ and r²¹ together they form an oxygen atom in the epoxy ring; and n is an integer of 1 or 2.

Particularly preferred tricyclic compounds (I) are, in addition to FK506, ascomycin derivatives such as halogenated-ascomycin (e.g. 33-epi-chloro-33-deoxyascomycin) which is disclosed in EP 427,680, example 66a.

Another preferred example of macrolites as immunosuppressants is rapamycin [THE MERCK INDEX (12th edition), No. 8288] and its derivatives. A preferred example of a derivative is an O-substituted derivative in which the hydroxy group at position 40 of formula A shown on page 1 of WO 95/16691, incorporated herein by reference, is replaced with OR1.

Wherein R 1 is hydroxyalkyl, hydroalkoxyalkyl, acylaminoalkyl, and aminoalkyl; e.g. 40-0- (2-hydroxy) ethyl rapamycin, 40-0- (3-hydroxy) propyl rapamycin, 40-0- [2- (2-hydroxy) ethoxy] ethyl rapamycin and 40-0- (2-acetaminoethyl) -rapamycin. Such O-substituted derivatives can be prepared by reacting rapamycin (or dihydro or deoxo-rapamycin) with an organic substituent attached to a leaving group (e.g. RX where R is an organic substituent that is desired as an O-substituent such as alkyl, allyl, or benzyl and X is a leaving group such as CCl3C (NH) O or CF3O3) under the appropriate reaction conditions. The reaction conditions may be acidic or neutral, e.g. in the presence of an acid such as trifluoromethanesulfonic acid, camphorsulfonic acid, p-toluenesulfonic acid, or their corresponding pyridinium or substituted pyridinium salts, wherein X is CCl3C (NH) O or in the presence of a base such as pyridine, substituted pyridine, diisopropylethylamine or pentamethylpiperidine, where X is means CF3SO3. 40-O- (2-hydroxy) ethyl rapamycin, which is disclosed in WO94 / 09010, a disclosure of which is hereby incorporated by reference, is particularly preferred. The tricyclic compounds (I) and rapamycin and its derivatives have a similar basic structure, i.e. a tricyclic macrolide structure, and at least one similar biological property (e.g., immunosuppressive activity).

The tricyclic compounds (I) and rapamycin and its derivatives may exist in the form of salts, which include conventional non-toxic and pharmaceutically acceptable salts such as a salt with inorganic or organic bases, in particular, alkali metal salts such as sodium and potassium salts, metal salt alkaline earths such as calcium and magnesium salt, ammonium salt and amine salt such as triethylamine salt and N-benzyl-N-methylamine.

With regard to the macrolide-tacrolimus compound or a hydrate thereof used in the present invention, it is to be understood that there may be conformers and one or more stereoisomers such as optical and geometric isomers due to asymmetric carbon atom (s) or double bond (s). and such conformers and isomers are also included within the scope of the macrolide compound. In addition, the macrolide compounds may exist in the form of a hydrate which is within the scope of the present invention.

One of the preferred specific examples of a slow release formulation according to the present invention is a formulation comprising a solid dispersion formulation wherein the macrolide compound is present in amorphous form in the solid medium exhibiting a T63.2 of 0.7 to 15 hours. Presence or absence of a diffraction peak detected by X-ray crystallographic analysis, thermal analysis, etc. shows whether the macrolide compound forms an amorphous phase or not in a solid dispersion composition.

Any pharmaceutically acceptable support capable of retaining the macrolide compound in the amorphous phase and which is solid at ambient temperature is satisfactory as a solid support for use in the solid dispersion composition mentioned above. Preferably, the solid base is a pharmaceutically acceptable water-soluble base; in particular, the substrate is, for example, one of the following water-soluble polymers:

PVC , polyethylene glycol (PEG) with an average molecular weight of 4000 or more, gelatin, etc.

Further, in use, the water-soluble polymers are individually used singly or in a mixture of two or more. A particularly preferred water-soluble support is a cellulose polymer or PVP; a particularly preferred water-soluble support is HPMC, PVP or a combination thereof. In particular, when low viscosity HPMC is used, it may have the more desirable slow release effect; an aqueous 2% solution of this type of HPMC with a viscosity of 1 to 4000 cps, preferably 1 to 50 cps, in particular 1 to 15 cps, measured at 20 ° C with a Brookfield viscometer, is preferable; in particular, HPMC 2910 with a viscosity of 3 cps (TC-5E, EW, Shinestu Chemical Co., Ltd.).

The weight ratio of the macrolide compound and such water-soluble base is preferably from 1: 0.05 to 1: 2, in particular from 1: 0.1 to 1: 1, especially from 1: 0.2 to 1: 0.4 .

The solid support may further be, for example, water-insoluble pharmaceutically acceptable supports capable of retaining the macrolide compound as an amorphous phase and being solids at ambient temperature. More specifically, the solid base includes, for example, waxes and water-insoluble polymers.

PL 193 244 B1

Particularly preferred examples of wax include glycerol monostearate and sucrose fatty acid esters (e.g., sucrose mono-, di- or triesters with medium to higher fatty acids, from 8 to 20 carbon atoms, e.g. caprylic, capric, lauric, myristic acid, palmitic, stearic, peanut, behenic, oleic, linolenic, etc.).

Other examples of wax include polyglycerin fatty acid ester. Any polyglycerin fatty acid ester, including monoester, diester or polyester polyglycerin with fatty acid, is satisfactory. Specific examples of polyglycerin fatty acid ester are hexa (tetra) glyceride behenate, mono (deca) glyceride caprylate, di (tri) glyceride caprylate, di (tri) glyceride caprate, mono (tetra) glyceride laurate, mono (hexate) laurate glyceride, mono (deca) glyceride laurate, mono (tetra) glyceride oleate, mono (hexa) glyceride oleate, mono (deca) glyceride oleate, di (tri) glyceride oleate, di (tetra) glyceride oleate, sesquiciride (tetra) oleate, glyceride oleate , penta (tetra) glyceride oleate, penta (hexa) glyceride oleate, deca (deca) glyceride oleate, mono (hepta) glyceride linoleate, di (tri) glyceride linolenate, di (tetra) glyceride linoleate, di (hexa) glyceride linolenate, mono (hexa) glyceride stearate, mono glyceride stearate mono (tetra) glyceride, mono (hexa) glyceride stearate, mono (deca) glyceride stearate, tri (tetra) glyceride stearate, tri (hexa) glyceride stearate, sesqui (hexa) glyceride stearate, perglyceride (glyceride tearide) stearate (hexa) glyceride, glyceride deca (deca) stearate, mono- (tetra) glyceride palmitate, mono (hexa) glyceride palmitate, tri (tetra) glyceride palmitate, tri (hexa) glyceride palmitate, sesqui (hexa) glyceride palmitate, glyceride (hexa) glyceride palmitate, glyceride (hexa) glyceride palmitate , penta (hexa) glyceride palmitate and deca (deca) glyceride palmitate.

Preferred polyglycerol fatty acid esters are e.g. hexa (tetra) glyceride behenate (e.g., under the trade name Poem J-46B, manufactured by Riken Vitamin Co., Ltd.), penta (tetra) glyceride stearate [e.g. trade name PS-310, manufactured by Sakamoto Yakuhin Kogyo Co., Ltd.], mono (tetra) glyceride stearate [e.g., trade name MS-310, manufactured by Sakamoto Yakuhin Kogyo Co., Ltd.], penta stearate (hexa) glyceride [tradename PS-500, manufactured by Sakamoto Yakuhin Kogyo Co., Ltd.], sesqui (hexa) glyceride stearate [tradename SS-500, manufactured by Sakamoto Yakuhin Kogyo Co., Ltd.], mono (deca) glyceride stearate, and mixtures thereof. Particularly preferred waxes are glycerol monostearate and low HLB fatty acid ester of sucrose [e.g. F-50, F-20, F-10, etc., manufactured by Dai-ichi Kogyo Seiyaku, Co., Ltd.].

The weight ratio of macrolide compound and wax is preferably from 1:10 to 1: 100, in particular from 1:40 to 1:60 if the wax is e.g. glycerol monostearate; their weight ratio is preferably from 1: 0.2 to 1:20, in particular 1: 0.5 to 1: 5 when the wax is e.g. a sucrose fatty acid ester; their weight ratio is preferably from 1: 0.1 to 1: 100, in particular 1: 0.5 to 1:50 if the wax is e.g. a polyglycerol fatty acid ester.

Preferred water-insoluble polymers include, e.g., ethyl cellulose, methacrylate copolymers (e.g., Eudragity such as Eudragit E, R, S, RS, LD, etc.). In a situation where the water-insoluble polymer is ethylcellulose, such a pharmaceutically acceptable substance can be used in the present invention. However, its preferred viscosity is from 3 to 110 cps, in particular from 6 to 49 cps, in particular from 9 to 11 cps, the viscosity of a 5% ethyl cellulosatoluene / ethanol solution (80:20) being measured according to the viscosity test described in: USP 23, NF18. For example, a preferred ethylcellulose is ETHOCELL (viscosity: 10) (tradename, Dow Chemical (USA)).

The weight ratio of the macrolide compound and the water-insoluble polymer is preferably from 1: 0.01 to 1:10, in particular from 1: 0.1 to 1: 5; in particular from 1: 0.1 to 1: 1 when the water-insoluble polymer is ethyl cellulose; the weight ratio of these components is in particular from 1: 0.5 to 1: 5 when the water-insoluble polymer is a methacrylate copolymer.

When preparing the solid dispersion composition of the present invention, the above-mentioned solid base, such as a water-soluble base and a water-insoluble base, may be used singly or in combination. When a water-insoluble base is used as the solid support of the present invention, a suitable dissolution profile for the solid dispersion composition can be achieved by mixing an appropriate amount of a water-soluble base, such as a water-soluble polymer (e.g., HPMC). If desired, suitable auxiliaries (lactose, etc.), binders, colorants, sweeteners, flavoring, diluents, antioxidants (vitamin E, etc.) are added to prepare a solid dispersion composition in the form of a solid dispersion. etc) and measures

Lubricants (e.g., synthetic aluminum silicate, magnesium stearate, calcium hydrogen phosphate, calcium stearate, talc, etc.).

In addition, depending on the nature of the solid support, the dissolution rate of the macrolide compound from the solid dispersion composition is sometimes too slow, or it is sometimes desirable to increase the initial rate of dissolution of the compound. In such a case, the dissolution rate of the macrolide compound from the solid dispersion composition may be adjusted by adding suitable disintegrants to the solid dispersion composition [e.g. sodium croscarmellose (CC-Na), calcium carboxymethyl cellulose (CM-Ca), low-substituent hydroxypropyl cellulose (L-HPC), sodium starch glycolate, microcrystalline cellulose, crospovidone, etc.] or suitable surfactants [e.g., hydrogenated polyoxyethylene oil castor, polyoxyl 40 stearate, polysorbate 80, sodium lauryl sulfate, sucrose fatty acid ester (HLB greater than 10), etc]. When the solid base is a water-soluble base, the solid dispersion composition preferably is substantially substantially free of a disintegrant when preparing a slow release formulation in accordance with the present invention.

The particle size of the solid dispersion composition in which the macrolide compound is present as an amorphous phase in the solid support is preferably equal to or less than 500 µm. In particular, the composition has a particle size passing through a 350 µm, especially 250 µm, sieve.

Furthermore, the solid dispersion composition of the macrolide compound contained in the slow release formulation of the invention can be obtained by the methods described in EP 0 240 773 and WO 91/19495 etc .; these methods are described in more detail below.

The macrolide compound is dissolved in an organic solvent (e.g., ethanol, dichloromethane or an aqueous mixture thereof, etc.), then an appropriate amount of the solid support is added and the resulting mixture is sufficiently dissolved or suspended or allowed to swell. Then the mixture is sufficiently kneaded. After removing the solvent from the mixture, the residue is dried, ground and reduced in particle size, thereby obtaining a solid dispersion composition wherein the macrolide compound is present as an amorphous phase in the solid base. During the kneading process, if necessary, lubricants such as calcium hydrogen phosphate, auxiliaries such as lactose, etc. can also be added.

A slow release formulation containing a macrolide-tacrolimus compound or a hydrate thereof of the present invention can also be prepared by using a finely divided powder of the macrolide compound. Particle size control of the macrolide compound can be achieved with grinding equipment routinely used in the pharmaceutical industry, such as a plug mill, impact mill, micronizer, ball mill (dry or wet), to name a few examples. The fine powder of the macrolide compound should have a particle diameter distribution in the range of 0.1-50 µm, preferably 0.2-20 µm, and in particular 0.5 ~ 10 µm, and / or an average diameter size of 0.2 ~ 20 µm, preferably 0.5-10 µm, and in particular 1 ~ 5 µm.

The solid dispersion composition and the finely divided powder of the macrolide compound, prepared according to the above methods, can be used as such in a slow release formulation. With regard to handling convenience, water dispersibility and oral dispersibility, the composition is particularly preferably formulated as a slow release formulation in the form of a powder, comminuted powder, granule, tablet or capsule by routine formulation techniques (e.g. molding). by pressing).

If necessary, then a slow release formulation can be prepared by mixing the composition as a solid dispersion or powdered macrolide compounds, e.g. with diluents or lubricants (such as sucrose, lactose, starch, crystalline cellulose, synthetic aluminum silicate, magnesium stearate, calcium stearate, calcium hydrogen phosphate, and talc) and / or colorants, sweeteners, flavors and disintegrators for standard applications. The resulting mixture is then thoroughly mixed to prepare a slow-release formulation. The slow release formulation or solid dispersion composition or particulate powder of the macrolide compound of the present invention may be pre-dispersed in water and juice for oral administration as a liquid preparation.

The effective dose of the macrolide compound will vary depending upon the nature of the compound, the age of the patient, his / her disease, the severity of the disease, or other factors. In general, the active ingredient is used at a dose of about 0.001 to 1000 mg, preferably 0.01 to 500 mg, in particular 0.1

To 100 mg per day for the therapeutic treatment of the disease; generally, the average single dose is about 0.01 mg, 0.1 mg, 0.5 mg, 1 mg, 5 mg, 10 mg, 50 mg, 100 mg, 250 mg, and 500 mg.

Following oral administration, the slow release formulation of the macrolide compound of the invention characteristically releases the macrolide compound in a sustained manner, and the pharmaceutical activity is sustained over a long period of time. In accordance with the present invention, the frequency of administration of macrolide compounds with pharmacological activity can be reduced. In particular, it has become possible to provide a macrolide-containing pharmaceutical preparation that can only be administered once a day. Moreover, it is now possible to provide a pharmaceutical composition free from the risk of undesirable effects due to temporary over-concentration and to ensure pharmacological efficacy over a sufficiently extended period of time.

The slow release formulation of the present invention is useful in the treatment and / or prevention of the following diseases and conditions due to the pharmacological activities exhibited by said macrolide compounds, especially the tricyclic compounds (I).

Rejection reactions by organ or tissue transplantation such as heart, kidney, liver, bone marrow, skin, cornea, lung, pancreas, small intestine, limbs, muscles, nerves, intervertebral disc, trachea, myoblast, cartilage, etc .; graft versus host reactions after bone marrow transplantation; autoimmune diseases such as rheumatoid arthritis, systemic lupus erythematosus, Hashimoto's thyroiditis, multiple sclerosis, severe myasthenia gravis, type I diabetes mellitus etc .; and infections caused by pathogenic microorganisms (e.g. Aspergillus fumigatus, Fusarium oxysporum, Trichophyton asteroides etc.).

Inflammatory or hyperproliferative skin diseases, or cutaneous manifestations of diseases mediated by immunology (e.g. psoriasis, contact atopic dermatitis, eczematoid dermatitis, seborrheic dermatitis, lichen planus, pemphigus, bullous pemphigoid, epidermal bullous, urticaria, angioedema, vasculitis , erythema, cutaneous eosinophilia, lupus erythematosus, acne and alopecia areata); autoimmune eye diseases (e.g. Keratoconjunctivitis, Vernal Conjunctivitis, Behcet's Disease Uveitis, Keratitis, Keratitis Herpetic, Conical Keratitis, Epithelial Keratitis, Endosperm, Ocular Premphygus, Mooren's Ulcer, Scleritis, Graves' Ophthalmopathy, Vogt's Syndrome -Koyanagi-Harada, keratoconjunctivitis (dry eye), pimple, iritis and ciliary body inflammation, sarcoidosis, endocrine ophthalmopathy, etc.); reversible obstructive airway diseases [asthma (e.g., bronchial asthma, allergic asthma, intrinsic asthma, extrinsic asthma, and dust asthma), especially chronic or long-term asthma (e.g. late asthma and airway hyperresponsiveness), bronchitis, etc.]; inflammation of the mucosa or vessels (e.g. gastric ulcer, ischemic or thrombotic vascular injury, ischemic bowel disease, enteritis, necrotizing enterocolitis, intestinal damage associated with thermal burns, leukotriene B4 mediated diseases); enteritis / allergies (e.g. celiac disease, proctitis, eosinophilic gastroenteritis, amastocytosis, Crohn's disease and ulcerative colitis); food-related allergic diseases with distant gastrointestinal symptoms (e.g. migraine, rhinitis and eczema); renal diseases (e.g., interstitial nephritis, Goodpasture's syndrome, hemolytic uremic syndrome, and diabetic nephropathy); nervous diseases (e.g. dermatomyositis, Guillain-Barr syndrome, Meniere's disease, polyneuritis, neuritis, cerebral infarction, Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis (ALS) and radiculography);

cerebral ischemic disease (e.g., head trauma, cerebral hemorrhage (e.g., subarachnoid hemorrhage, intracerebral hemorrhage), cerebral thrombosis, cerebral embolism, cardiac arrest, stroke, transient ischemic attack (TLA), hypertensive encephalopathy, cerebral infarction); endocrine diseases (e.g., hyperthyroidism and Basedow's disease); blood diseases (e.g. pure red cell aplasia, aplastic anemia, hypoplastic anemia, Werlhof's disease, autoimmune hemolytic anemia, agranulocytosis, pernicious anemia, megaloblast anemia and anerythroplasia); bone diseases (e.g., osteoporosis); respiratory diseases (e.g., sarcoidosis, pulmonary fibrosis, and idiopathic interstitial pneumonia); skin diseases (e.g. dermatomyositis, common leukoderma, ichthyosis vulgaris, photosensitivity, and cutaneous T cell lymphoma); circulatory diseases (e.g. hardening of arteries, atherosclerosis, aortic syndrome, polyarteritis nodosa, and myocardosis); collagen diseases (e.g., scleroderma, Wegener's granulomatosis, and Sjogren's syndrome); obesity; eosinophilic fasciitis;

Periodontal diseases (e.g. damage to the gingiva, periodontium, alveolar bone or substantia osseadentis); nephrotic syndrome (e.g., glomerulonephritis); male pattern baldness, senile baldness; muscular dystrophy; pyoderma and Sezary's syndrome; chromosomal abnormalities (e.g., Down's syndrome); addison's disease; active oxygen mediated diseases [e.g. organ trauma (e.g. ischemic related organ disorders (e.g. heart, liver, kidney, gastrointestinal tract, etc.) associated with storage, transplantation, or ischemic diseases (e.g. thrombosis, cardiac infarction, etc.)): bowel diseases (e.g. endotoxin shock, pseudomembranous colitis, and drug-induced colitis or radiation): kidney diseases (e.g. ischemic acute renal failure, chronic renal failure): lung diseases (e.g. oxygen toxicity in the lungs or drugs (e.g. paracort, bleomycin, etc.), lung cancer and emphysema): eye diseases (e.g. cataracts, iron (siderosis bulbi), retinitis, pigmentosis, senile plaques, vitreous scarring, alkaline corneal burns): dermatitis (e.g. erythema multiforme linear immunoglobulin A dermatitis bullous, basal substance dermatitis): and other diseases (e.g. gingivitis, periodontitis, sepsis, pancreatitis and diseases caused by environmental pollution (eg air pollution), aging, carcinogen, cancer metastasis and air pressure depression syndrome)]; diseases caused by the release of histamine or leukotriene C4; restenosis of the coronary arteries after angioplasty and prevention of postoperative adhesions; autoimmune diseases and inflammation (e.g. primary mucosal edema, autoimmune atrophic gastritis, premature menopause, male infertility, juvenile diabetes mellitus, pemphigus vulgaris, pemphigoid, sympathetic eye or conjunctivitis, lens-induced uveitis, idiopathic leukopenia, active chronic hepatitis, idiopathic cirrhosis, thyroid cirrhosis lupus erythematosus, autoimmune orchitis, arthritis (e.g. arthritis deformans) or polychondritis); HIV (HIV) infection, AIDS; allergic conjunctivitis; hypertrophic scar and keloids from trauma, burns, or surgery.

Moreover, said tricyclic macrolides have liver regenerating activity and / or activity of stimulating hepatocyte hypertrophy and hyperplasia. Thus, the pharmaceutical composition of the present invention is useful for enhancing the treatment effect and / or prophylaxis of liver diseases [e.g. immunogenic diseases (e.g. chronic autoimmune liver disease such as autoimmune liver disease, primary biliary cirrhosis or sclerosing cholangitis), partial liver resection, acute liver necrosis (e.g., toxin necrosis, viral hepatitis, shock or anoxia), hepatitis B, non-A non-B hepatitis, cirrhosis and hepatic failure (e.g. fulminant hepatitis, late-onset hepatitis and "acute-chronic" hepatic failure (acute hepatic failure associated with chronic liver disease))].

Moreover, the present composition is also useful for increasing the prevention and / or treatment effect of various diseases due to the useful pharmacological activity of said tricyclic macrolides, such as chemotherapeutic effect enhancing activity, cytomegalovirus infection activity, anti-inflammatory activity, peptidyl-prolyl isomerase or rotamase inhibitory activity, antimalarial activity, antitumor activity, etc.

The present invention further provides a method of performing a dissolution test for a solid formulation comprising a macrolide compound that uses a test solution containing an appropriate amount of a cellulose polymer. In general, a dissolution test to test the release characteristics of a therapeutically active ingredient dissolved from a solid preparation containing that ingredient is performed according to the dissolution test, method 2 (paddle method, 50 rpm), JP XIII, or the dissolution test of USP 23 , NF1B or in the European Pharmacopoeia (3rd edition). However, when carrying out a dissolution test as with a formulation containing a small amount of the macrolide compound, the release of the macrolide compound based on its intrinsic content may not reach 100% even after several hours. This is because when the amount of the macrolide compound is small, adsorption of the macrolide compound on the surfaces of the sample vessel, filter, etc. will have an effect of increasing the size.

After much research, the present inventors have determined that by adding an appropriate amount of a cellulose polymer (such as HPMC, hydroxypropyl cellulose phthalate, MC, CMC-Na, hydroxyethyl cellulose, hydroxypropyl cellulose (HPC), etc.) to the test solution and, if necessary, adding phosphoric acid (or similar) into the test solution to bring the pH to a value not greater than 7 and to avoid the adverse effect of increasing the pH on the stability of the macrolide compound, can inhibit

The effect of adsorption of the macrolide compound on the surfaces of the test jig to achieve a recovery rate of substantially 100%. The preferred cellulose polymer is hydroxypropyl cellulose or its equivalent, the preferred viscosity of which is that when 5.0 g is dissolved in 95 ml of water, and after centrifugation to remove foam where necessary, the viscosity of the solution is measured using a rotary viscometer. at 25 0.1 ° C, the solution has a viscosity of 75-150 cps. e.g. hydroxypropyl cellulose with an average molecular weight of about 100,000, such as that available from Aldrich, meets these requirements.

The "appropriate amount" of cellulose polymer to be added to the test solution is 0.001 ~ 0.1%, preferably 0.002 ~ 0.01%, most preferably 0.005%, all values ​​relating to the total amount of the test solution.

The dissolution test, method 2 (paddle method), JP XIII, and the dissolution test in USP 23, NF18 or in the European Pharmacopoeia (3rd edition) are well known methods for testing the release kinetics of an active ingredient from a solid pharmaceutical product. These are dissolution tests using the specified vessel, stirrer, and other means, controlling the amount of test solution, temperature of the test solution, rotational speed, and other conditions.

Where necessary, the test is performed with a test solution adjusted to a suitable pH. In the present invention, the pH is preferably not higher than 7. In the present invention, "dissolution test method 2 (paddle method, 50 rpm), JP XIII" means "dissolution test method 2 (agitator method). vane), JP XIII, which is performed using agitation (50 rpm). The corresponding descriptions in JP XIII, USP 23 (NF18) and the European Pharmacopoeia (3rd edition) are incorporated herein by reference.

The invention will now be described in the following examples, but not limited thereto. In the following examples, FK506 is admixed as a monohydrate in the preparation of compositions containing this material, although the amount is expressed as weight of FK506 and not as monohydrate.

Example 1

FK506 1.0 mg

HPMC 2910 1.0 mg total 2.0 mg

FK506 was dissolved in ethanol, and HPMC 2910 was added to the resulting solution to allow sufficient swelling of the FK506. Thereafter, the mixture was kneaded together. The resulting kneaded mixture was transferred to a stainless tray, dried under reduced pressure and ground using a coffee grinder. Thereafter, the obtained powder was subjected to particle size reduction by the following processes to produce a solid dispersion composition (hereinafter referred to as SDC) 1-1) to 1-6).

(1) the comminuted powder is passed through a 250-µm sieve and its fraction remaining on the sieve is determined by SDC 1-1) (&gt; 250 µm).

(2) The fraction passed through the sieve in process (1) was passed through a 180-µm sieve and its fraction remaining on the sieve was designated SDC 1-2) (180-250 µl ι).

(3) The fraction passed through the sieve in process (2) was passed through a 150-µL sieve and its fraction remaining on the sieve was designated SDC 1-3) (150-180 µm).

(9) The fraction passed through the sieve in process (3) was passed through a 106-µm sieve and its fraction remaining on the sieve was designated SDC 1-4) (106-150 µl).

(5) The fraction passed through the sieve in process (4) was passed through a 75-µτη sieve and its fraction remaining on the sieve was designated SDC 1-5) (75-106 µm).

(6) The fraction passed through the sieve in process (5) is designated SDC 1-6) (&lt;75 µm).

Example 2

SDC 1-2), which was prepared in Example 1, was sufficiently mixed with lactose (58.0 mg) and the resulting mixture was encapsulated to form a capsule.

Example 3

In a manner similar to that of Example 1, a particulate powder with the following SDCs with a particle size of 180 to 250 µm was produced.

PL 193 244 B1

<td>SDC</td><td>Macrolide compound</td><td>Water-soluble base</td>

<td> 3-1)</td><td>FK506 (1.0 mg)</td><td>HPMC 2910 (0.3 mg)</td>

<td> 3-2)</td><td>FK506 (1.0 mg)</td><td>HPMC 2910 (0.1 mg)</td>

In addition, SDC 3-1) was sufficiently mixed with lactose (58.7 mg) and the resulting mixture was encapsulated to form a capsule 3-1). SDC 3-2) was sufficiently mixed with lactose (58.9 mg) and the resulting mixture was encapsulated to form a 3-2 capsule).

Example 4

In a similar manner to SDC 1-2) of Example 1, the following SDCs were prepared.

<td>SDC</td><td>Macrolide compound</td><td>Water-soluble base</td>

<td> 4-1)</td><td>FK506</td><td>MC</td>

<td>(2, 0 mg in total)</td><td>(1.0 mg)</td><td>(1.0 mg)</td>

<td> 4-2)</td><td>FK506</td><td>PVP</td>

<td>(2.0 mg in total)</td><td>(1.0 mg)</td><td>(1/0 mg)</td>

<td> 4-3)</td><td>FK506</td><td>HPMC 2910</td>

<td>(2, 0 mg in total)</td><td>(1/0 mg)</td><td>(1/0 mg)</td>

<td> 4-4)</td><td>FK506</td><td>HPC</td>

<td>(2.0 mg in total)</td><td>(1.0 mg)</td><td>(1.0 mg)</td>

<td> 4-5)</td><td>FK506</td><td>PEG</td>

<td>(2, 0 mg in total)</td><td>(1/0 mg)</td><td>(1/0 mg)</td>

<td> 4-6)</td><td>FK506</td><td>HPMC 2910</td>

<td>(2.0 mg in total)</td><td>(1/0 mg)</td><td>(0.8mg) PVP (0.2mg)</td>

In a similar manner to Example 2, lactose (in the appropriate amount) and magnesium stearate (0.6 mg) were added to the respective SDCs to make the corresponding capsules, each containing 60.0 mg in total.

Example 5

In a similar manner to SDC 1-2) in Example 1, SDC was prepared using FK506 (1.0 mg) and HPMC 2910 (1.0 mg).

Then, in a similar manner to Example 2, the following additives were respectively added to the SDC to make capsules 5-1) to 5-4), each containing 60.0 mg in total.

<td>Capsule No.</td><td>Appendix (extras)</td><td></td>

<td> 5-1)</td><td>crystalline cellulose, magnesium stearate</td><td>(right amount) (0.6 mg)</td>

<td> 5-2)</td><td>calcium hydrogen phosphate magnesium stearate</td><td>(right amount) (0.6 mg)</td>

<td> 5-3)</td><td>lactose L-HPC magnesium stearate</td><td>(right amount) (3.0 mg) (0.6 mg)</td>

<td> 5-4)</td><td>maize starch calcium stearate</td><td>(right amount) (0.6 mg</td>

Example 6

FK506 1.0 g

HPMC 2910_0.3 g

Total 1.3 g

FK506 was dissolved in ethanol and HPMC 2910 was added to the resulting solution to allow sufficient swelling. Thereafter, the mixture was kneaded. The resulting kneaded material was transferred to a stainless tray, dried under vacuum and ground using a coffee grinder.

PL 193 244 B1

Thereafter, the resulting powder was subjected to size reduction by the following processes to prepare SDC 6-1) to 6-6).

(1) The comminuted powder was passed through a 250 µm sieve, and its fraction remaining on the sieve was designated SDC 6-1) (&gt; 250 µm 2).

(2) the fraction which passed through the sieve in the process (1) was passed through a 180 µm sieve and its fraction remaining on the sieve was designated as SDC 6-2) (180-250 µ ^ ι), (3) the fraction passed through the sieve in process (2), 150 µm was passed through a sieve and its fraction remaining on the sieve was designated as SDC 6-3) (150-180 µ ^ ι), (4) the fraction that passed through the sieve in process (3), was passed through the sieve 106 ^ m and its fraction remaining on the sieve was marked as SDC 6-4) (106-150 μ ^ ι), (5) fraction, which passed through the sieve in the process (4) was passed through a sieve 75 µm and its fraction remaining on the sieve was designated as SDC 6-5) (75-106 µ ^ ι), (6) the fraction that passed through the sieve in the process (5) labeled SDC 6-6).

Example 7

SDC 6-4) (1.3 mg) which was obtained in Example 6 was mixed thoroughly with lactose (58.1 mg) and magnesium stearate (0.6 mg) and the resulting mixture was filled into capsules which were designated capsule 7.

Example 8

In a similar manner to Example 1, the following SDCs were prepared with a particle size of 180-250 µm.

<td>SDC</td><td>Macrolide compound</td><td>Water-soluble base</td>

<td> 8-1)</td><td>ascomycin (1.0 mg)</td><td>HPMC 2910 (0.3 mg)</td>

<td> 8-2)</td><td>33-epi-chloro-33-deoxyascomycin (1.0 mg)</td><td>HPMC 2910 (0.3 mg)</td>

<td> 8-3)</td><td>40-0- (2-hydroxy) ethyl rapamycin (1.0 mg)</td><td>HPMC 2910 (0.3 mg)</td>

In a similar manner to Example 7, each capsule was prepared by adding lactose (58.1 mg) and magnesium stearate (0.6 mg).

Example 9

SDC 9

FK506 10g

HPMC 2910 3 g

Calcium hydrogen phosphate_3 g

A total of 16 g

Preparation 9

SDC 9 16 g

Lactose qs

Stearate_7 g

Total 700 g

FK506 was dissolved in ethanol, and HPMC 2910 was added to the resulting solution and mixed sufficiently, then further added calcium hydrogen phosphate. After drying in vacuo overnight, the resulting mixture was subjected to size reduction using a speed mill and a roller granulator; the resulting powder was sieved with a sieve of 212 µm; its fraction passing through the sieve was designated SDC 9. SDC 9, lactose and magnesium stearate were blended together to make formulation 9. Formulation 9 was filled into capsule No. 1 in an amount of 350 mg and into a gelatin capsule No. 5 in an amount of 70 mg, which are designated Formulations A and B, respectively.

PL 193 244 B1

Example 10

SDC 10

FK506 10 g

HPMC 2910 3 g

Lactose_3 g

A total of 16 g

Preparation 10

SDC 10 16 g

Lactose qs

Magnesium stearate_7 g

Total 700g

In a similar manner to Example 9, SDC 10 and Formulation 10 were prepared respectively. Example 11

SDC 11

FK506 10 g

HPMC 2910 3 g

Calcium hydrogen phosphate_3 g

A total of 16 g

Preparation 11

SDC 11 16 g

Lactose qs

Magnesium stearate_7 g

Total 700 g

FK506 was dissolved in ethanol, HPMC 2910 was added and sufficiently mixed with the resulting solution, then additional calcium hydrogen phosphate was added. After the resulting mixture was dried in vacuo overnight, the mixture was subjected to size reduction using a speed mill and a roller granulator; the resulting powder was sieved through a 250 µm sieve and a 180 µm sieve; the 180-250 µ ^ ι fraction was defined as SDC 11. SDC 11, lactose and magnesium stearate were mixed together to make formulation 11. Formulation 11 was filled into a 350 mg capsule No. 1 and a 70 mg amount into gelatin capsule No. 5, which are designated Formulations C and D, respectively.

Example 12

SDC 12

FK506 2 g

Glycerin Monostearate 98 g

HPMC 2910_20 g

Total 120 g

Preparation 12

SDC 12 120 g

Magnesium stearate_1,2

A total of 121.2 g

Glycerin monostearate was heated, melted at 80 ° C, and FK506 was added to it (stirring to dissolve). HPMC 2910 was added to the resulting mixture for sufficient mixing and the resulting mixture was then transferred to a tray for self-cooling. The solids obtained by cooling were ground using a coffee grinder and then sieved through a 500 µm sieve. Its fraction passing through the sieve was designated SDC 12. SDC 12 was mixed with

PL 193 244 B1

<td colspan="2">magnesium stearate to make formulation 12 which was then filled into capsule No. 5 in a quantity of 60.6 mg. The resulting capsule was designated formulation E.</td>

<td>Example 13</td><td></td>

<td>SDC 13 FK506</td><td>2 g</td>

<td>Methacrylate-aminoalkyl copolymer (Eudragit RL)</td><td>6 g</td>

<td>Calcium hydrogen phosphate</td><td>2 g</td>

<td>Overall</td><td>10 g</td>

<td>Preparation 13 SDC 13</td><td>10 g</td>

<td>Lactose</td><td>130 g</td>

<td>Overall</td><td>140 g</td>

<td colspan="2">FK506 and methacrylate aminoalkyl copolymer were dissolved in ethanol, and then</td>

<td colspan="2">Calcium phosphate and the resulting mixture were sufficiently mixed. The mixture was dried overnight under reduced pressure, ground in a mortar, and sorted using 150 μm and 106 μm sieves,</td>

<td>to generate the 106-150 µ ^ ι fraction as SDC 13.</td><td>SDC 13 was mixed with lactose to make Formulation 13,</td>

<td colspan="2">and then filled in an amount of 70 mg into a gelatin capsule No. 5 prepared as preparation F.</td>

<td>Example 14 SDC 14 FK506</td><td>2 g</td>

<td>Methacrylate-aminoalkyl copolymer (Eudragit RL)</td><td>4.6 g</td>

<td>Methacrylate-aminoalkyl copolymer (Eudragit RS)</td><td>1.4 g</td>

<td>Calcium hydrogen phosphate</td><td>2 g</td>

<td>Overall</td><td>10 g</td>

<td>Preparation 14 SDC 14</td><td>10 g</td>

<td>Lactose</td><td>130 g</td>

<td>Overall</td><td>140 g</td>

<td colspan="2">In a similar manner to Example 13, SDC 14 was produced with a particle size of 106-</td>

<td colspan="2">150 μm and formulation 14. Formulation 14 was then filled in an amount of 70 mg into a gelatin capsule No. 5</td>

<td>prepared as preparation of G. Example 15</td><td></td>

<td colspan="2">SDC 15</td>

<td>FK506</td><td>2 g</td>

<td>Methacrylate-aminoalkyl copolymer (Eudragit RL)</td><td>3 g</td>

<td>Methacrylate-aminoalkyl copolymer (Eudragit RS)</td><td>3 g</td>

<td>Calcium hydrogen phosphate</td><td>2 g</td>

<td>Overall</td><td>10 g</td>

<td colspan="2">Preparation 15</td>

<td>SDC 15</td><td>10 g</td>

<td>Lactose</td><td>130 g</td>

<td>Overall</td><td>140 g</td>

PL 193 244 B1

In a similar manner as in Example 13, SDC 15 was prepared with a particle size of 106-150 μτπ and formulation 15. The formulation 15 was then filled into a 70 mg gelatin capsule No. 5 prepared as Formulation H.

Example 16

<td colspan="2">SDC 16</td>

<td>FK506</td><td>2 g</td>

<td>Ethylcellulose</td><td>0.4 g</td>

<td>Lactose</td><td>6 g</td>

<td>Overall</td><td>8.4 g</td>

<td colspan="2">Preparation 16</td>

<td>SDC 16</td><td>8.4 g</td>

<td>Lactose</td><td>131.6 g</td>

<td>Overall</td><td>140 g</td>

<td colspan="2">FK506 and ethyl cellulose were dissolved in ethanol, then lactose was added and the resulting mixture was mixed sufficiently. The mixture was dried in vacuo overnight, ground in a mortar and sorted using 150 µτπ and 106 µ ^ ι sieves to produce a 106-150 µm fraction. as SDC 16. SDC 16 was mixed with lactose and prepared as formulation 16 and then filled with 70 mg of gelatin capsule No. 5 prepared as formulation I. Example 17</td>

<td>SDC 17 FK506 Ethylcellulose Lactose</td><td>2 g 1 g 6 g</td>

<td>Overall</td><td>9 g</td>

<td>Preparation 17 SDC 17 Lactose</td><td>9 g 131 g</td>

<td>Overall</td><td>140 g</td>

<td colspan="2">In a similar manner as in Example 16, SDC 17 was prepared with a particle size of 106-150 μτη and formulation 17. Then formulation 17 was filled in a 70 mg amount into a gelatin capsule No. 5 prepared</td>

<td colspan="2">as a preparation of J.</td>

<td>Example 18 SDC 18 FK506 Ethylcellulose Hydroxypropyl methylcellulose Lactose</td><td>2 g 0.4 g 0.6 g 6 g</td>

<td>Overall</td><td>9 g</td>

<td>Preparation 18</td><td></td>

<td>SDC 18</td><td>9 g</td>

<td>Lactose</td><td>131 g</td>

<td>Overall</td><td>140 g</td>

In a similar manner to Example 16, SDC 18 was prepared with a particle size of 106-150 μτπ and formulation 18. Formulation 18 was then filled into a 70 mg gelatin capsule No. 5 prepared as Formulation K.

PL 193 244 B1

Example 19

SDC 19

FK506 2 g

Ethylcellulose 0.6 g

HPMC 2910 0.6 g

Lactose_6 g

Total 9.2 g

Preparation 19

SDC 19 9.2 g

Lactose_130.8 g

A total of 140 g

In a similar manner to Example 16, SDC 19 was prepared with a particle size of 106-150 μm and formulation 19. Then formulation 19 was filled in a 70 mg amount into a gelatin capsule No. 5 prepared as Formulation L.

Example 20

SDC 20

FK506 10g

Ethylcellulose 3 g

MPMC 2910 3 g

Lactose_50 g

Total 66 g

Preparation 20

SDC 20 66g

Lactose qs

Magnesium stearate_7fl

Total 700g

FK506 was dissolved in ethanol and added to it, and ethyl cellulose was dissolved. HPMC 2910 and lactose were sufficiently mixed with the resulting solution. After drying in vacuo overnight, the resulting mixture was subjected to size reduction using a power mill and a roller granulator; the resulting powder was sieved with a 250 µm sieve; its fraction passing through the sieve was determined by SDC 20. SDC 20, lactose and magnesium stearate were blended together to make formulation 20. Formulation 20 was filled into capsule No. 1 in an amount of 350 mg and into a gelatin capsule No. 5 in an amount of 70 mg, which were designated as Formulations M and N, respectively.

Example 21

SDC 21

<td>FK506</td><td>10 g</td>

<td>Ethylcellulose</td><td>3 g</td>

<td>HPMC 2910</td><td>3 g</td>

<td>Lactose</td><td>20 g</td>

<td>Overall</td><td>36 g</td>

<td>Preparation 21</td><td></td>

<td>SDC 21</td><td>36 g</td>

<td>Lactose</td><td>qs</td>

<td>Magnesium stearate</td><td>7 g</td>

<td>Overall</td><td>700 g</td>

PL 193 244 B1

In a manner similar to that in Example 20, its fraction passing through the 212 μτη sieve denoted as SDC 21 and formulation 21 were prepared. Then 350 mg was filled into gelatin capsule No. 1 with 350 mg and 70 mg with 70 mg gelatin capsule No. 5 prepared as preparations O and P , respectively.

Example 22

SDC 22

FK506 1 g

Sucrose ester 1 g fatty acid (HLB = 6) (DK F-50 ester)

Total 2 g

Preparation 22

SDC 22 2 g

Lactose_68 g

Total 70 g

FK506 was dissolved in ethanol / acetone (1/1). After heating the solution to 75 ° C, the sucrose fatty acid ester was added to dissolve, followed by cooling at room temperature. The mixture was dried under vacuum overnight, ground in a mortar and sorted using 150 µm and 106 µm sieves to produce a 106-150 µm fraction as SDC 22. SDC 22 was mixed with lactose to make formulation 22. It was then filled in an amount of 70 mg into a gelatin capsule No. 5 prepared as preparation Q.

Example 23

SDC 23

FK506 1 g

Fatty acid ester and 0.75 g of sucrose (HLB = 6) (DK F-50 ester)

Sucrose fatty acid ester (HLB = 2) 0.25 g

Total 2 g

Preparation 23

SDC 23 2 g

Lactose_68 g

Total 70 g

In a similar manner as in Example 22, SDC 13 was prepared at a particle size of 106-150 µm and formulation 23. Formulation 23 was then filled into a 70 mg gelatin capsule No. 5 prepared as Formulation R.

Example 24

SDC 24

FK506 1 g

Fatty acid ester and 1 g of sucrose (HLB = 1) (DK F-10 ester)

Lactose_1 g

A total of 3 g

Preparation 24

SDC 24 3 g

Lactose_67 g

Total 70 g

In a similar manner as in Example 22, SDC 24 was prepared with a particle size of 106-150 µm and formulation 24. Formulation 24 was then filled into a 70 mg gelatin capsule No. 5 prepared as Formulation S.

PL 193 244 B1

Example 25

SDC 25

FK506 1 g

Sucrose fatty acid ester (HLB = 1) (DK F-10 ester) 1 g

Lactose_3 g

A total of 5 g

Preparation 24

SDC 24 5 g

Lactose_65 g

Total 70 g

In a similar manner as in Example 22, SDC 25 was prepared with a particle size of 106-150 µm and formulation 25. Preparation 25 was then filled into a 70 mg gelatin capsule No. 5 prepared as Formulation T.

Example 26

SDC 26

FK5Q6 1g

Fatty acid ester and 1 g of sucrose (HLB = 1) (DK F-10 ester)

Lactose_5 g

A total of 7 g

Preparation 26

SDC 26 7 g

Lactose_63 g

Total 70 g

In a similar manner to Example 22, SDC 26 was prepared at a particle size of 106-150 µm and formulation 26. Formulation 26 was then filled into a 70 mg gelatin capsule No. 5 prepared as Formulation U.

Example 27

SDC 27

FK506 1 g

A fatty acid ester and 30 g of tetraglycerin

Lactose_15 g

Total 46 g

Preparation 27

SDC 27 46 g

Lactose_24 g

Total 70 g

The tetraglycerin fatty acid ester melted by heating at 80 ° C was added and FK506 was dissolved with stirring. Lactose was added to the resulting mixture, mixed and then spontaneously cooled in a tray.

The resulting solid was ground using a coffee grinder, and sorted using 150 μm and 106 μm sieves to produce a 106-150 μm fraction as SDC 27. SDC 27 was mixed with lactose and prepared as formulation 27, then formulation 27 was filled at 70 mg. gelatin capsule No. 5 prepared as preparation V.

PL 193 244 B1

Example 28

SDC 28

FK506 1 g

A fatty acid ester and 30 g of tetraglycerin

Polysorbate_0.3 g

A total of 31.3 g

Preparation 28

SDC 28 31.3 g

Lactose_38.7 g

Total 70 g

In a similar manner as in Example 27, SDC 28 was prepared with a particle size of 106-150 µm and formulation 28. Formulation 28 was then filled into a 70 mg gelatin capsule No. 5 prepared as Formulation W.

Example 29

SDC 29

FK506 1 g

A fatty acid ester and 1 g of tetraglycerin

Lactose_3 g

A total of 5 g

Preparation 29

SDC 29 5 g

Lactose_65 g

Total 70 g

Ethanol was added to the tetraglycerin fatty acid ester. The resulting mixture was melted by heating at 40 ° C, FK506 was added and then melted with stirring. Lactose was added to the resulting mixture, mixed and then cooled spontaneously in a tray. The resulting solid was ground using a coffee grinder, dried in vacuo overnight, and sorted using 150 µm and 106 µm sieves to produce a 106-150 µm fraction as SDC 29. SDC 29 was mixed with lactose and prepared as Formulation 29, and Formulation 29 was then filled with 70 mg into a gelatin capsule No. 5, prepared as Formulation X.

Example 30

Preparation 30

FK506 - crushed powder 0.5 g

Lactose 29.2 g

Magnesium stearate_0.3

A total of 30 g

Crystalline FK506 was ground using a jet mill and mixed with lactose and magnesium stearate to form formulation 30. Formulation 30 was then filled into a 60 mg gelatin capsule No. 5 prepared as formulation Z. The particle size range of FK506 crushed powder after the jet mill was 1-10 μm and the average particle size was approximately 3 μm.

Example 31

Dissolution Test

Sample for sample:

(1) Formulations A and C which were prepared in the aforementioned examples.

(2) Control formulation (rapid release formulation) which is a 1 mg capsule formulation with the following ingredients. It was prepared in a similar manner to Examples 1 and 2 of WO

PL 193 244 B1

91/19495, by mixing components (e) and (f) with a solid dispersion composition consisting of the following components (a) to (d) and by coating.

(a) tacrolimus (FK506) 1 mg (b) hydroxypropyl methylcellulose 1 mg (c) lactose 2 mg (d) croscarmellose sodium 1 mg (e) lactose 59.35 mg (f) magnesium stearate 0.65 mg

Test method:

The test was carried out according to Japanese Pharmacopoeia, 13th Edition, Dissolution Test No. 2 (paddle method, 50 rpm) using an aqueous 0.005% hydroxypropyl cellulose adjusted to pH 4.5 as the test solution. The data obtained are presented below.

<td>Time (hours)</td><td>Preparation A (%)</td><td>Time (hours)</td><td>Preparation C (%)</td>

<td> 0</td><td> 0,0</td><td> 0</td><td> 0,0</td>

<td> 0,5</td><td> 17,4</td><td> 1</td><td> 12,1</td>

<td> 1</td><td> 35,6</td><td> 2</td><td> 30,9</td>

<td> 2</td><td> 57,6</td><td> 4</td><td> 55,9</td>

<td> 3</td><td> 71,9</td><td> 6</td><td> 71,3</td>

<td> 4</td><td> 80,9</td><td> 8</td><td> 81,6</td>

<td> 6</td><td> 89,7</td><td> 10</td><td> 87,0</td>

<td> 9</td><td> 95,2</td><td> 12</td><td> 90,4</td>

<td>Time (hours)</td><td>Control sample (%)</td>

<td> 0</td><td> 0,0</td>

<td> 0,17</td><td> 30,1</td>

<td> 0,5</td><td> 68,4</td>

<td> 1</td><td> 92,8</td>

<td> 2</td><td> 100,1</td>

Example 32

In a similar manner to Example 31, a dissolution test was performed. As a result, various parameters were obtained by calculation in the Weibull function and T63.2%.

Score

<td>Preparation</td><td>Dmax (%)</td><td>m</td><td>n</td><td>Ti</td><td>T63.2% (h)</td>

<td> 1</td><td> 2</td><td> 3</td><td> 4</td><td> 5</td><td> 6</td>

<td>Capsule 7</td><td> 101,7</td><td> 2,69</td><td> 1,18</td><td> 0,0</td><td> 2,3</td>

<td>AND</td><td> 95,9</td><td> 2,24</td><td> 1,03</td><td> 0,0</td><td> 2,2</td>

<td>C.</td><td> 92,5</td><td> 6,14</td><td> 1,24</td><td> 0,0</td><td> 4,3</td>

<td>E.</td><td> 101,6</td><td> 1, 93</td><td> 0,60</td><td> 0,0</td><td> 3,0</td>

<td>F.</td><td> 95,6</td><td> 2,51</td><td> 1,00</td><td> 0,0</td><td> 2,5</td>

<td>G.</td><td> 99,0</td><td> 3,69</td><td> 0,91</td><td> 0,0</td><td> 4,2</td>

<td>H.</td><td> 88,8</td><td> 6,34</td><td> 0,88</td><td> 0,0</td><td> 8,2</td>

<td>L.</td><td> 95,6</td><td> 2,51</td><td> 1,00</td><td> 0,0</td><td> 2,5</td>

<td>J.</td><td> 99,0</td><td> 3,69</td><td> 0,91</td><td> 0,0</td><td> 4,2</td>

<td>K.</td><td> 101,2</td><td> 1,69</td><td> 0,80</td><td> 0,0</td><td> 1,9</td>

The table continues

<td> 1</td><td> 2</td><td> 3</td><td> 4</td><td> 5</td><td> 6</td>

<td>L.</td><td> 91,4</td><td> 2,48</td><td> 0,75</td><td> 0,0</td><td> 3,3</td>

<td>M.</td><td> 90,4</td><td> 1,61</td><td> 0,62</td><td> 0,0</td><td> 2,1</td>

<td> 0</td><td> 83,9</td><td> 2,5</td><td> 0,67</td><td> 0,0</td><td> 3,9</td>

<td>Q</td><td> 104,7</td><td> 1,89</td><td> 0,93</td><td> 0,0</td><td> 2,0</td>

<td>R</td><td> 92,1</td><td> 2,09</td><td> 0,82</td><td> 0,0</td><td> 2,5</td>

<td>S.</td><td> 86,0</td><td> 3,73</td><td> 0,89</td><td> 0,0</td><td> 4,4</td>

<td>T.</td><td> 87,9</td><td> 2,00</td><td> 0,93</td><td> 0,0</td><td> 2,1</td>

<td>AT</td><td> 93,4</td><td> 1,03</td><td> 0,86</td><td> 0,0</td><td> 1,0</td>

<td>V</td><td> 83,6</td><td> 1,14</td><td> 0,54</td><td> 0,0</td><td> 1,3</td>

<td>IN</td><td> 87,1</td><td> 1,30</td><td> 0,69</td><td> 0,0</td><td> 1,5</td>

<td>WITH</td><td> 85,7</td><td> 1,98</td><td> 0,75</td><td> 0,0</td><td> 2,5</td>

<td>Control sample</td><td> 100,9</td><td> 0,41</td><td> 1,10</td><td> 0,0</td><td> 0,4</td>

Example 33

Oral absorbability

Sample for trials:

(1) Formulations B and D which were prepared in the aforementioned examples.

(2) Control slide (same as control slide in example 31)

Test method:

The test samples were orally administered to 6 cynomological monkeys (FK506 dose: 1 mg / monkey) to test the blood concentration of FK506 after administration. Seventeen hours prior to administration, the feeds were removed from the feed table for cynomological monkeys weighing approximately 6 kg. The animals were then fasted for up to 12 hours after dosing. Before initiation of sample administration and thereafter, water was administered ad libitum. When the samples were administered, the animals were simultaneously treated with water (20 ml). At predetermined time intervals after dosing, 1 ml of blood was drawn from a forearm vein using a sterile syringe in a plastic tube containing heparin and stored at -80 ° C until initiation of the drug trial. The total concentration of the drug FK506 in the blood was tested by immunoassay with FK506-specific enzyme (EIA) known in JP-A-1-92659, the description of which is cited and included in the description of the application.

Average value

<td>Time (hours)</td><td>Preparation B</td><td>Preparation D</td><td>Control sample</td>

<td> 1</td><td> 2</td><td> 3</td><td> 4</td>

<td> 0</td><td> 0,00</td><td> 0,00</td><td> 0,00</td>

<td> 0,5</td><td> 0,44</td><td> 0,28</td><td> 0,91</td>

<td> 1</td><td> 2,59</td><td> 1,03</td><td> 3,02</td>

<td> 2</td><td> 4,26</td><td> 2,27</td><td> 7,13</td>

<td> 4</td><td> 3,89</td><td> 3,14</td><td> 3,27</td>

<td> 1</td><td> 2</td><td> 3</td><td> 4</td>

<td> 6</td><td> 3,48</td><td> 4,42</td><td> 3,85</td>

<td> 8</td><td> 3,47</td><td> 4,12</td><td> 2,63</td>

<td> 10</td><td> 3,70</td><td> 4,06</td><td> 2,48</td>

<td> 12</td><td> 3,73</td><td> 4,10</td><td> 2,51</td>

<td> 19</td><td> 3,85</td><td> 4,13</td><td> 2,27</td>

<td> 16</td><td> 3,60</td><td> 4,75</td><td> 2,20</td>

<td> 18</td><td> 2,96</td><td> 3,95</td><td> 1,76</td>

<td> 24</td><td> 2,21</td><td> 2,57</td><td> 1,32</td>

PL 193 244 B1

The maximum concentration in the blood (Cmax) is defined as the maximum content of the drug in the whole blood. Tmax is the time required to reach the maximum blood concentration. MRT is defined as the mean retention time. The area under the blood concentration-time curve (AUC) was calculated by the trapezoidal method. As an index of change in absorbability, CV (standard deviation / mean in%) was calculated.

Test results

<td>Samples for testing (Preparation No.)</td><td>Cmax (ng / mL) (CV (%))</td><td>Tmax (h) (CV (%))</td><td>MRT (h) (CV (%))</td><td>AUC (ngh / mL) (CV (%))</td>

<td>B</td><td> 5,51±1,02</td><td> 8,2±2,9</td><td> 21,1±0,5</td><td> 126,3±22,</td>

<td></td><td> (45,4)</td><td> (87,8)</td><td> (5,5)</td><td> (43,1)</td>

<td>D</td><td> 5,48±0,94</td><td> 10,0±2,7</td><td> 22,6±1,0</td><td> 144,3121,0</td>

<td></td><td> (41,8)</td><td> (66,9)</td><td> (11,2)</td><td> (35,7)</td>

<td>A sample</td><td> 8,41±1,46</td><td> 3,3±0,8</td><td> 17,6±0,9</td><td> 91,1±20,4</td>

<td>control</td><td> (42,6)</td><td> (62,2)</td><td> (12,7)</td><td> (54,9)</td>

Example 34

In a manner similar to that in Example 33, the oral absorbability of the various formulations of the present invention was determined.

Results

<td>Samples for testing (Preparation No.)</td><td>Cmax (ng / mL) (CV (%))</td><td>Tmax (h) (CV (%))</td><td>MRT (h) (CV (%))</td><td>AUC (ngh / mL) (CV (%))</td>

<td>E.</td><td> 9,36±1,08</td><td> 6,3±1,7</td><td> 20,0±0,4</td><td> 186,6±18,5</td>

<td></td><td> [28,4]</td><td> [67,5]</td><td> [5,1]</td><td> [24,3]</td>

<td>L.</td><td> 6,16±0,57</td><td> 4,3±1,1</td><td> 19,3±0,5</td><td> 135,5±17,7</td>

<td></td><td> [22,6]</td><td> [61,4]</td><td> [6,9]</td><td> [31,9]</td>

<td>Q</td><td> 4,70±0,39</td><td> 5,0±1,7</td><td> 21,4±1,6</td><td> 122,6±10,2</td>

<td></td><td> [20,2]</td><td> [83,0]</td><td> [7,0]</td><td> [20,3]</td>

<td>WITH</td><td> 5,72±0,92</td><td> 8,0±1,2</td><td> 20,9±1,2</td><td> 133,2±16,1</td>

<td></td><td> [39,3)]</td><td> [35,4)</td><td> [13,7]</td><td> [29,6]</td>

<td>A sample</td><td> 12,27±2,60</td><td> 1,4±0,3</td><td> 14,3±1,0</td><td> 80,8±15,1</td>

<td>control</td><td> [51,8]</td><td> [46,5]</td><td> [17,7]</td><td> [45,8]</td>

The above results show that the formulations used in the above trials, when administered orally, exhibit lower Cmax, sufficiently long Tmax and MRT than the rapid release (control) formulation. Compared to the rapid release formulation, the AUC values ​​of the above formulations are nearly the same or greater; or the above sustained release formulations give less variation of C max and / or AUC in subjects as compared to the sustained release formulation. In accordance with the invention of the present application, the small change in individuals of the maximum blood concentration or the area under the blood concentration-time curve after oral administration of a macrolide compound as compared to a rapid release formulation can be determined using the index of change in blood absorbability of the macrolide compound, viz. standard deviation / mean (CV in%) of the maximum blood concentration or area under the blood concentration-time curve. The term "minor change" means its small CV value; in particular, the term means that the CV value is less than that of the rapid release formulation described above.