Stabilized pharmaceutical composition in lyophilized form

This application is a divisional of U.S. Ser. No. 09/786,125, filed Mar. 1, 2001, now U.S. Pat. No. 6,774,104, which is the national-stage under 35 U.S.C. §371 of PCT/JP00/04381, filed Jun. 29, 2000. This application also claims priority to JAPAN 11/187713, filed Jul. 1, 1999.

The present invention relates to a stabilized pharmaceutical composition in lyophilized form containing a cyclic polypeptide compound. More particularly, the present invention relates to a stabilized pharmaceutical composition in lyophilized form containing a cyclic polypeptide compound or its pharmaceutically acceptable salt and a stabilizer.

The cyclic polypeptide compound of the present invention is represented by the general formula (I):

wherein R 1 is a hydrogen atom or an acyl group and R 2 and R 3 are, the same or different, a hydrogen atom or a hydroxyl group. The compound has an antimicrobial activity, particularly an antifungal activity and a β-1,3-glucan synthase inhibiting action, and is useful for preventing and treating various kinds of infectious diseases including Pneumocystis carinii infection, e.g., carinii pneumonia.

Among the cyclic polypeptide compounds represented by the above formula (I), a compound wherein R 1 is a hydrogen atom and R 2 and R 3 are hydroxyl groups and a compound wherein R 1 , R 2 and R 3 are hydrogen atoms are obtained by a fermentation process disclosed by European Patent No. 0462531 and processes disclosed by WO97/32975 and by WO97/47738. A compound wherein R 1 is an acyl group and its production process are disclosed by U.S. Pat. Nos. 5,376,634 and 5,569,646 and WO96/11210 and WO99/40108.

The cyclic polypeptide compounds (I) and their salts are generally unstable to light, humidity, acids, heat and the like. Therefore, desired is development of pharmaceutical preparations in which the cyclic polypeptide compounds and their salts are stabilized.

The present invention provides a stabilized pharmaceutical composition in lyophilized form containing a cyclic polypeptide compound (I) or its pharmaceutically acceptable salt and a stabilizer.

The “acyl group” for R 1 in the formula (I) representing the cyclic polypeptide compound of the present invention is now explained. In the context of the present specification, “lower” means having one to six carbon atoms unless otherwise indicated.

As examples of the acyl group, may be mentioned aliphatic acyl groups, aromatic acyl groups, aromatic-aliphatic acyl groups and heterocyclic acyl groups derived from aliphatic, aromatic, aromatic-aliphatic and heterocyclic carboxylic acids.

Examples of the aliphatic acyl groups include lower or higher alkanoyl groups such as formyl, acetyl, propanoyl, butanoyl, 2-methylpropanoyl, pentanoyl, 2,2-dimethylpropanoyl, hexanoyl, heptanoyl, octanoyl, nonanoyl, decanoyl, undecanoyl, dodecanoyl, tridecanoyl, tetradecanoyl, pentadecanoyl, hexadecanoyl, heptadecanoyl, octadecanoyl, nonadecanoyl, icosanoyl, etc.; cycloalkanoyl groups such as cyclopentanoyl and cyclohexanoyl; lower alkoxycarbonyl groups such as methoxycarbonyl, ethoxycarbonyl, t-butoxycarbonyl, t-pentyloxycarbonyl, heptyloxycarbonyl, etc.; lower alkanesulfonyl groups such as methanesulfonyl, ethanesulfonyl, etc.; lower alkoxysulfonyl groups such as methoxysulfonyl, ethoxysulfonyl, etc.; and the like.

Examples of the aromatic acyl groups include aroyl groups such as benzoyl, toluoyl, naphthoyl and the like.

Examples of the aromatic-aliphatic acyl groups include ar(lower)alkanoyl groups such as phenyl(C 1 –C 6 )alkanoyl (e.g., phenylacetyl, phenylpropanoyl, phenylbutanoyl, phenylisobutanoyl, phenylpentanoyl, phenylhexanoyl, etc.), naphthyl(C 1 –C 6 )alkanoyl (e.g., naphthylacetyl, naphthylpropanoyl, naphthylbutanoyl, etc.) and the like; ar(lower)alkenoyl group such as phenyl(C 3 –C 6 )alkenoyl (e.g., phenylpropenoyl, phenylbutenoyl, phenylmethacryloyl, phenylpentenoyl, phenylhexenoyl, etc.), naphthyl(C 3 –C 6 )alkenoyl (e.g., naphthylpropenoyl, naphthylbutenoyl, etc. and the like;

ar (lower)alkoxycarbonyl groups such as phenyl(C 1 –C 6 )alkoxycarbonyl (e.g., benzyloxycarbonyl, etc.), fluorenyl(C 1 –C 6 )alkoxycarbonyl (e.g., fluorenylmethoxycarbonyl, etc.) and the like; aryloxycarbonyl groups such as phenoxycarbonyl, naphthoxycarbonyl, etc.; aryloxy(lower)alkanoyl groups such as phenoxyacetyl, phenoxypropionyl, etc.; arylcarbamoyl groups such as phenylcarbamoyl, etc.; arylthiocarbamoyl groups such as phenylthiocarbamoyl, etc.; arylglyoxyloyl groups such as phenylglyoxyloyl, naphthylglyoxyloyl, etc.; arylsulfonyl groups which may be optionally substituted by a lower alkyl group such as phenylsulfonyl, p-tolylsulfonyl, etc.; and the like.

Examples of the heterocyclic acyl groups include heterocyclic carbonyl groups such as thenoyl, furoyl, nicotinoyl, etc.;

heterocyclic(lower)alkanoyl groups such as heterocyclic acetyl, heterocyclic propanoyl, heterocyclic butanoyl, heterocyclic pentanoyl, heterocyclic hexanoyl, etc.; heterocyclic(lower)alkenoyl groups such as heterocyclic propenoyl, heterocyclic butenoyl, heterocyclic pentenoyl, heterocyclic hexenoyl, etc.; heterocyclic glyoxyloyl and the like.

The acyl group for R 1 may have one or more suitable substituent(s). Among the above-mentioned examples for the acyl groups, an aroyl group which may have one or more suitable substituent(s) is particularly preferable.

Examples of suitable substituents in the acyl group include a heterocyclic group substituted by an aryl group having a lower alkoxy group, a heterocyclic group substituted by an aryl group having a lower alkoxy(lower)alkoxy group, a heterocyclic group substituted by an aryl group having a lower alkoxy(higher)alkoxy group, a heterocyclic group substituted by an aryl group having a cyclo(lower)alkyloxy group, a heterocyclic group substituted by an aryl group having a heterocyclic group, a heterocyclic group substituted by a cyclo(lower)alkyl group having a cyclo(lower)alkyl group, a heterocyclic group substituted by an aryl group having an aryl group substituted by a lower alkoxy(lower)alkoxy and a heterocyclic group substituted by an aryl group having a heterocyclic group substituted by a cyclo(lower)alkyl group.

Among these examples, preferred are an unsaturated 3- to 8-membered heteromonocyclic group containing one to two oxygen atom(s) and one to three nitrogen atom(s) and substituted by phenyl having (C 4 –C 6 )alkoxy, an unsaturated condensed heterocyclic group containing one to two sulfur atom(s) and one to three nitrogen atom(s) and substituted by phenyl having (C 4 –C 6 )alkoxy, an unsaturated 3- to 8-membered heteromonocyclic group containing one to two sulfur atom(s) and one to three nitrogen atom(s) and substituted by phenyl having (C 1 –C 4 )alkoxy(C 4 –C 6 )alkoxy, an unsaturated 3- to 8-membered heteromonocyclic group containing one to two sulfur atom(s) and one to three nitrogen atom(s) and substituted by phenyl having (C 1 –C 4 )alkoxy(C 7 –C 14 )alkoxy, a saturated 3- to 8-membered heteromonocyclic group containing one to four nitrogen atom(s) and substituted by phenyl having (C 1 –C 4 )alkoxy(C 7 –C 14 )alkoxy, an unsaturated condensed heterocyclic group containing one to two sulfur atom(s) and one to three nitrogen atom(s) and substituted by phenyl having cyclo(C 4 –C 6 )alkyloxy, an unsaturated condensed heterocyclic group containing one to two sulfur atom(s) and one to three nitrogen atom(s) and substituted by phenyl, a saturated 3- to 8-membered heteromonocyclic group containing one to two oxygen atom(s) and one to three nitrogen atom(s), a saturated 3- to 8-membered heteromonocyclic group having one to four nitrogen atom(s) and substituted by cyclo(C 4 –C 6 )alkyl having cyclo(C 4 –C 6 )alkyl, an unsaturated 3- to 8-membered heteromonocyclic group having one to two sulfur atom(s) and one to three nitrogen atom(s) and substituted by phenyl having phenyl substituted by (C 1 –C 4 )alkoxy(C 1 –C 4 )alkoxy, an unsaturated 3- to 8-membered heteromonocyclic group containing one to two sulfur atom(s) and one to three nitrogen atom(s) and substituted by phenyl having a saturated 3- to 8-membered heteromonocyclic group which contains one to four nitrogen atom(s) and is substituted by cyclo(C 4 –C 6 )alkyl, and an unsaturated condensed heterocyclic group containing one to two sulfur atom(s) and one to three nitrogen atom(s) and substituted by phenyl having a saturated 3- to 8-membered heteromonocyclic group which contains one to four nitrogen atom(s) and has cyclo(C 4 –C 6 )alkyl.

Among these, particularly preferred are an isoxazolyl group substituted by phenyl having pentyloxy, an imidazothiadiazolyl group substituted by phenyl having pentyloxy, a thiadiazolyl group substituted by phenyl having methoxyhexyloxy, a thiadiazolyl group substituted by phenyl having methoxyoctyloxy, a thiadiazolyl group substituted by phenyl having methoxyheptyloxy, an imidazothiadiazolyl group substituted by phenyl having cyclohexyloxy, an imidazothiadiazolyl group substituted by phenyl having dimethylmorpholino, a piperazinyl group substituted by phenyl having methoxyheptyloxy, a piperazinyl group substituted by phenyl having methoxyoctyloxy, a piperazinyl group substituted by cyclohexyl having cyclohexyl, a thiadiazolyl group substituted by phenyl having phenyl substituted by methoxyethoxy, a thiadiazolyl group substituted by phenyl having phenyl substituted by methoxybutoxy, a thiadiazolyl group substituted by phenyl having phenyl substituted by ethoxypropoxy, an imidazothiadiazolyl group substituted by phenyl having piperazinyl substituted by cyclohexyl, an imidazothiadiazolyl group substituted by phenyl having piperazinyl substituted by cyclohexyl, and the like.

Accordingly, particularly suitable examples of the acyl group of R 1 may be a benzoyl group having isoxazolyl substituted by phenyl having pentyloxy, a benzoyl group having imidazothiadiazolyl substituted by phenyl having pentyloxy, a benzoyl group having thiadiazolyl substituted by phenyl having methoxyhexyloxy, a benzoyl group having thiadiazolyl substituted by phenyl having methoxyoctyloxy, a benzoyl group having thiadiazolyl substituted by phenyl having methoxyheptyloxy, a benzoyl group having imidazothiadiazolyl substituted by phenyl having cyclohexyloxy, a benzoyl group having imidazothiadiazolyl substituted by phenyl having dimethylmorpholino, a benzoyl group having piperazinyl substituted by phenyl having methoxyheptyloxy, a benzoyl group having piperazinyl substituted by phenyl having methoxyoctyloxy, a benzoyl group having piperazinyl substituted by cyclohexyl having cyclohexyl, a benzoyl group having thiadiazolyl substituted by phenyl having phenyl substituted by methoxyethoxy, a benzoyl group having thiadiazolyl substituted by phenyl having phenyl substituted by methoxybutoxy, a benzoyl group having thiadiazolyl substituted by phenyl having phenyl substituted by ethoxypropoxy, a benzoyl group having imidazothiadiazolyl substituted by phenyl having piperazinyl substituted by cyclohexyl, a benzoyl group having imidazothiadiazolyl substituted by phenyl having piperazinyl substituted by cyclohexyl, and the like.

Particularly preferable examples of the acyl groups of R 1 are represented by the formulas:

The cyclic polypeptide compounds (I) having the above-mentioned acyl groups may be prepared from a compound having a hydrogen atom as R 1 and hydroxyl groups as R 2 and R 3 or a compound having hydrogen atoms as R 1 , R 2 and R 3 according to the U.S. Pat. Nos. 5,376,634 and 5,569,646 and WO96/11210 and WO99/40108.

Suitable salts of the cyclic polypeptide compounds (I) are soluble in water and pharmaceutically acceptable salts including salts with bases and acid addition salts. Such a salt may be prepared by treating the cyclic polypeptide compound (I) with an appropriate base or acid according to the conventional method.

As salts with bases, may be mentioned salts with inorganic bases such as alkali metal salts (e.g., sodium salts, potassium salts, etc.), alkaline earth metal salts (e.g., calcium salts, magnesium salts, etc.), ammonium salts and the like; salts with organic bases such as organic amine salts (e.g., triethylamine salts, diisopropylethylamine salts, pyridine salts, picoline salts, ethanolamine salts, triethanolamine salts, dicyclohexylamine salts, N,N′-dibenzylethylenediamine salts, etc.); and the like.

As acid addition salts, may be mentioned inorganic acid addition salts (e.g., hydrochlorides, hydrobromides, sulfates, phosphates, etc.); and organic carboxylic or sulfonic acid addition salts (e.g., formates, acetates, trifluoroacetates, maleates, tartrates, fumarates, methnesulfonates, benzenesulfonates, toluenesulfonates, etc.). Further, may also be mentioned salts with basic or acidic amino acids (e.g., salts with arginine, aspartic acid, glutamic acid, etc.).

The cyclic polypeptide compounds (I) of the present invention also include possible conformers and a pair or more of stereoisomers such as geometric isomers and optical isomers which may exist due to asymmetric carbon atoms.

The preferable ones of the cyclic polypeptide compounds (I) are represented by the following formulas (II) to (VI):

(to be continued on the next page)

The most preferable one is represented by the formula (II).

The amount of the cyclic polypeptide compound (I) or its pharmaceutically acceptable salt contained in the composition for a single unit dosage of the present invention is 0.1 to 400 mg, more preferably 1 to 200 mg, still more preferably 10 to 100 mg, specifically 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 70, 75, 80, 85, 90, 95 and 100 mg.

As the stabilizer, may be mentioned polysaccharides, disaccharides, sodium chloride and a combination thereof.

Examples of the polysaccharide are dextran, starch, cellulose and hyaluronic acid; and examples of the disacharide are lactose, maltose and sucrose. The polysaccharide or disaccharide contained in the pharmaceutical composition of the present invention may be α-monohydrate, α-anhydride, β-anhydride or a combination thereof.

The amount of the stabilizer used in the pharmaceutical composition of the present invention should be at least sufficient for stabilizing the cyclic polypeptide compound (I) or its pharmaceutically acceptable salt in the composition. In order to stabilize the cyclic polypeptide compound (I), one part by weight of the stabilizer with respect to one part by weight of the cyclic polypeptide compound (I) or its pharmaceutically acceptable salt in the present composition is sufficient at least. The stabilizer may also serve as a carrier or an excipient. Thus the use amount of stabilizer does not have a particular upper limit and may be determined in consideration of the weight or volume of the composition with respect to a unit dose of the compound and the like. However, such amount is preferably 0.4 to 50 parts by weight, more preferably 0.6 to 20 parts by weight, still more preferably 0.8 to 10 parts by weight with respect to one part by weight of the cyclic polypeptide compound (I) or its pharmaceutically acceptable salt, though it varies depending upon the kind and the used amount of the cyclic polypeptide compound (I) or its pharmaceutically acceptable salt, its preparation form and/or the like. Specifically, it is more preferable that 1 to 20 parts, still more preferably 2 to 10 parts by weight of the disaccharide are used with respect to one part by weight of the cyclic polypeptide compound (I) or its pharmaceutically acceptable salt. Specifically, it is more preferable that 0.6 to 20 parts, still more preferably 0.8 to 10 parts by weight of sodium chloride are used with respect to one part by weight of the cyclic polypeptide compound (I) or its pharmaceutically acceptable salt.

The pharmaceutical composition of the present invention may be produced according to methods known in the art with using additives if necessary. Here, Basic Lecture on Development of Pharmaceuticals XI 20 Production of Pharmaceuticals (the second volume) (edited by Kyosuke Tsuda and Hisashi Nogami and published by Chizyo Shoten) is mentioned for reference. The lyophilized composition may be obtained by preparing an aqueous solution of the cyclic polypeptide compound (I) or its pharmaceutically acceptable salt and the stabilizer, optionally adding a pH adjustor (citric acid anhydrous, sodium hydroxide, etc.) as required to attain pH 4.0–7.5, preferably pH 4.5–7.0, and then lyophilizing the resulting solution in vial according to a conventional method. Thus, the stabilized pharmaceutical composition in lyophilized form, when dissolved in purified water, preferably gives a solution of pH 4.0 to 7.5, more preferably pH 4.5 to 7.0. It is preferable that the thus prepared composition in lyophilized form is sealed and stored with shading. The lyophilized composition can be loaded in each vial in the solution form before lyophilizing or in lyophilized powder form after lyophilizing.

Since the cyclic polypeptide compound is not satisfactorily stable to humidity, it is necessary that the lyophilized composition of the present invention contains 3.4% by weight or less of water, preferably 3.0%, more preferably 2.0%.

Usually the stabilized pharmaceutical composition in lyophilized form is dissolved in isotonic sodium chloride solution as required and used as an injection solution. The pharmaceutical composition of the present invention may be used as an injection preparation which requires some compounding before use.

The present invention is now described in further detail by way of examples and test examples, which should not be construed to limit the scope of the invention. In the examples, the compounds of formula (II) to (VI) are referred to as Compounds (II) to (VI), respectively.