HK1080724B - Stabilized pharmaceutical composition in lyophilized form - Google Patents

Description

Stable pharmaceutical composition in lyophilized form

The application is a divisional application with the application number of 00801216.4, the application date of 29/6/2000 and the same name as above.

Technical Field

The present invention relates to a stable pharmaceutical composition comprising a cyclic polypeptide compound in lyophilized form. More particularly, the present invention relates to stable pharmaceutical compositions comprising a cyclic polypeptide compound or a pharmaceutically acceptable salt thereof and a stabilizing agent in lyophilized form.

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

wherein R is¹Is a hydrogen atom or an acyl group, and R²And R³Identical or different and is a hydrogen atom or a hydroxyl group. The compounds have antimicrobial activity, especially antifungal activity, and beta-1, 3-glucan synthase inhibitory activity, and are useful for the prevention and treatment of various infectious diseases, including pneumocystis carinii infections, such as pneumonia carinii (pneumonia carinii).

Background

In the cyclic polypeptide compound represented by the above formula (I), by the fermentation method disclosed in European patent No. 0462531 and the methods disclosed in WO97/32975 and WO97/47738, there is obtained a cyclic polypeptide compound wherein R is¹Is a hydrogen atom and R²And R³A compound which is hydroxy and wherein R¹、R²And R³A compound which is a hydrogen atom. U.S. Pat. Nos. 5,376,634 and 5,569,646 and WO96/11210 and WO99/40108 disclose compounds in which R is¹A compound which is an acyl group and a method for producing the same.

The cyclic polypeptide compound (I) and salts thereof are generally unstable to light, humidity, acid, heat, etc. therefore, there is a need for the development of pharmaceutical preparations in which the cyclic polypeptide compound and salts thereof are stabilized.

Disclosure of the invention

The present invention provides stable pharmaceutical compositions in lyophilized form comprising a cyclic polypeptide compound (I) or a pharmaceutically acceptable salt thereof and a stabilizer.

Now, explanation will be given of R in the formula (I) representing the cyclic polypeptide compound of the present invention¹The "acyl group" of (1). In the present specification, "lower" means having 1 to 6 carbon atoms unless otherwise specified.

As examples of acyl groups, mention may be made of aliphatic, aromatic, araliphatic and heterocyclic acyl groups derived from aliphatic, aromatic, araliphatic and heterocyclic carboxylic acids.

Examples of the aliphatic acyl group include lower or higher alkanoyl groups such as formyl, acetyl, propionyl, butyryl, 2-methylpropionyl, valeryl, 2-dimethylpropionyl, hexanoyl, heptanoyl, octanoyl, nonanoyl, decanoyl, undecanoyl, dodecanoyl, tridecanoyl, tetradecanoyl, pentadecanoyl, hexadecanoyl, heptadecanoyl, octadecanoyl, nonadecanoyl, eicosanoyl and the like; cycloalkanoyl groups such as cyclopentanyl and cyclohexanoyl; lower alkoxycarbonyl such as methoxycarbonyl, ethoxycarbonyl, tert-butoxycarbonyl, tert-pentyloxycarbonyl, heptyloxy and the like; lower alkanesulfonyl groups such as methanesulfonyl, ethanesulfonyl and the like; lower alkoxysulfonyl such as methoxysulfonyl, ethoxysulfonyl and the like; and the like.

Examples of the aromatic acyl group include aromatic acyl groups such as benzoyl, toluyl, naphthoyl and the like.

Examples of araliphatic acyl groups include ar (lower) alkanoyl groups, such as phenyl (C)₁-C₆) Alkanoyl (e.g., phenylacetyl, phenylpropionyl, phenylbutyryl, phenylpentyl, phenylhexanoyl, etc.), naphthyl (C)₁-C₆) Alkanoyl (e.g., naphthylacetyl, naphthylpropionyl, naphthylbutyryl, etc.), and the like;

ar (lower) alkenoyl, e.g. phenyl (C)₃-C₆) Alkanoyl (e.g., phenylpropanoyl, phenylbutenoyl, benzylacryloyl, phenylpentyloxyl, phenylhexenoyl, etc.), naphthyl (C)₃-C₆) Alkenoyl (e.g., naphthoyl, etc.), etc.;

aryl (lower) alkoxycarbonyl, such as phenyl (C)₁-C₆) Alkoxycarbonyl (e.g., benzyloxycarbonyl), fluorenyl (C)₁-C₆) Alkoxycarbonyl (e.g., fluorenylmethoxycarbonyl, etc.), etc.;

aryloxycarbonyl such as phenoxycarbonyl, naphthyloxycarbonyl, and the like;

aryloxy (lower) alkanoyl such as phenoxyacetyl, phenoxypropionyl, and the like;

arylcarbamoyl groups such as phenylcarbamoyl group and the like;

arylthiocarbamoyl such as phenylthiocarbamoyl and the like;

arylglyoxyloyl such as phenylglyoxyloyl, naphthylglyoxyloyl and the like;

arylsulfonyl groups which may be optionally substituted with a lower alkyl group such as benzenesulfonyl, p-toluenesulfonyl and the like; and the like.

Examples of heterocyclic acyl groups include heterocyclic carbonyl groups such as thenoyl, furoyl, nicotinyl, and the like; heterocyclic (lower) alkanoyl such as heterocyclic acetyl, heterocyclic propionyl, heterocyclic butyryl, heterocyclic valeryl, heterocyclic hexanoyl and the like;

heterocyclic (lower) alkenoyl such as heterocyclic acryloyl, heterocyclic crotonyl, heterocyclic pentenoyl, heterocyclic hexenoyl and the like;

heterocyclic glyoxyl, and the like.

R¹The acyl group of (a) may have one or more suitable substituents. Among the above-mentioned examples of the acyl group, an aroyl group which may have one or more suitable substituents is particularly preferred.

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

In these examples, it preferably contains 1 to 2 oxygen atoms and 1 to 3 nitrogen atoms and is substituted with (C)₄-C₆) Alkoxy-substituted phenyl-substituted unsaturated 3-8 membered heteromonocyclic group; containing 1-2 sulfur atoms and 1-3 nitrogen atoms and being provided with (C)₄-C₆) An alkoxy-substituted phenyl-substituted unsaturated fused heterocyclic group; containing 1-2 sulfur atoms and 1-3 nitrogen atoms and being provided with (C)₁-C₄) Alkoxy (C)₄-C₆) Alkoxy-substituted phenyl-substituted unsaturated 3-8 membered heteromonocyclic group; containing 1-2 sulfur atoms and 1-3 nitrogen atoms and being provided with (C)₁-C₄) Alkoxy (C)₇-C₁₄) Alkoxy-substituted phenyl-substituted unsaturated 3-8 membered heteromonocyclic group; containing 1 to 4 nitrogen atoms and being provided with (C)₁-C₄) Alkoxy (C)₇-C₁₄) Phenyl-substituted saturated 3-to 8-membered heteromonocyclic group of alkoxy group containing 1 to 2 sulfur atoms and 1 to 3 nitrogen atoms and being substituted with a ring (C)₄-C₆) An alkoxy-substituted phenyl-substituted unsaturated fused heteromonocyclic group; an unsaturated fused heterocyclic group containing 1 to 2 sulfur atoms and 1 to 3 nitrogen atoms and substituted with a phenyl group, a saturated 3-to 8-membered heteromonocyclic group containing 1 to 2 oxygen atoms and 1 to 3 nitrogen atoms; having 1 to 4 nitrogen atoms and being provided with a ring (C)₄-C₆) Ring of alkyl (C)₄-C₆) An alkyl-substituted saturated 3-8 membered heteromonocyclic group; containing 1-2 sulfur atoms and 1-3 nitrogen atoms and being substituted with (C)₁-C₄) Alkoxy (C)₁-C₄) Phenyl-substituted unsaturated 3-8 membered heteromonocyclic group of alkoxy-substituted phenyl; an unsaturated 3-to 8-membered heteromonocyclic group containing 1 to 2 sulfur atoms and 1 to 3 nitrogen atoms and substituted with a phenyl group having a saturated 3-to 8-membered heteromonocyclic group, wherein the saturated 3-to 8-membered heteromonocyclic group contains 1 to 4 nitrogen atoms and is substituted with a ring (C)₄-C₆) Alkyl substitution; an unsaturated condensed heterocyclic group containing 1 to 2 sulfur atoms and 1 to 3 nitrogen atoms and substituted with a phenyl group having a saturated 3-to 8-membered heteromonocyclic group containing 1 to 4 nitrogen atoms and having a ring (C)₄-C₆) An alkyl group.

Among these examples, particular preference is given to: isoxazolyl substituted with phenyl having pentyloxy group, imidazothiadiazolyl substituted with phenyl having pentyloxy group, thiadiazolyl substituted with phenyl having methoxyhexyloxy group, thiadiazolyl substituted with phenyl having methoxyoctyloxy group, thiadiazolyl substituted with phenyl having methoxyheptyloxy group, imidazothiadiazolyl substituted with phenyl having cyclohexyloxy group, imidazothiadiazolyl substituted with phenyl having dimethylmorpholino group, piperazinyl substituted with phenyl having methoxyheptyloxy group, piperazinyl substituted with phenyl having methoxyoctyloxy group, piperazinyl substituted with cyclohexyl group, thiadiazolyl substituted with phenyl having phenyl substituted with methoxyethoxy group, thiadiazolyl substituted with phenyl having phenyl substituted with methoxybutoxy group, thiadiazolyl substituted with phenyl having phenyl substituted with ethoxypropoxy group, An imidazothiadiazolyl group substituted with a phenyl group having a piperazinyl group substituted with a cyclohexyl group, and the like.

Thus, R¹Particularly suitable examples of acyl groups of (a) may be: benzoyl having an isoxazolyl, wherein the isoxazolyl is substituted with a phenyl having a pentyloxy group; benzoyl having an imidazothiadiazolyl group, wherein said imidazothiadiazolyl group is substituted with a phenyl group having a pentyloxy group; benzoyl having a thiadiazolyl group, wherein said thiadiazolyl group is substituted with a phenyl group having a methoxy group; benzoyl having a thiadiazolyl group, wherein said thiadiazolyl group is substituted with a phenyl group having a methoxyoctyloxy group; benzoyl having a thiadiazolyl group, wherein said thiadiazolyl group is substituted with a phenyl group having a methoxy heptyloxy group; benzoyl having an imidazothiadiazolyl group, wherein said imidazothiadiazolyl group is substituted with a phenyl group having a cyclohexyloxy group; benzoyl having an imidazothiadiazolyl group, wherein said imidazothiadiazolyl group is substituted with a phenyl group having a dimethylmorpholino group; benzoyl having a piperazinyl group, wherein the piperazinyl group is substituted with a phenyl group having a methoxy heptyloxy group; phenylmethyl with piperazinylAcyl, wherein the piperazinyl is substituted with phenyl having methoxyoctyloxy; benzoyl having a piperazinyl group, wherein the piperazinyl group is substituted with a cyclohexyl group having a cyclohexyl group; benzoyl having a thiadiazolyl group substituted with a phenyl group, wherein the phenyl group has a phenyl group substituted with a methoxyethoxy group; benzoyl having a thiadiazolyl group substituted with a phenyl group, wherein said phenyl group has a phenyl group substituted with a methoxybutoxy group; benzoyl having a thiadiazolyl group substituted with a phenyl group, wherein said phenyl group has a phenyl group substituted with ethoxypropoxy group; benzoyl having an imidazothiadiazolyl group substituted with a phenyl group, wherein said phenyl group has a piperidinyl group substituted with a cyclohexyl group; benzoyl having imidazothiadiazolyl substituted with phenyl, wherein said phenyl has piperazinyl substituted with cyclohexyl, and the like.

R¹A particularly preferred example of the acyl group of (a) is represented by the following formula:

according to U.S. Pat. Nos. 5,376,634 and 5,569,646 and WO96/11210 and WO99/40108, a hydrogen atom is substituted for R¹And hydroxy as R²And R³Or a compound having a hydrogen atom as R¹、R²And R³The cyclic polypeptide compound (I) having the acyl group can be produced.

Suitable salts of the cyclic polypeptide compounds (I) are soluble in water, and pharmaceutically acceptable salts include salts with base and acid addition salts. Such salts can be prepared according to conventional methods by treating the cyclic polypeptide compound (I) with a suitable base or acid.

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

As the acid addition salt, inorganic acid addition salts (e.g., hydrochloride, hydrobromide, sulfate, phosphate, etc.); and organic carboxylic or sulfonic acid addition salts (e.g., formates, acetates, trifluoroacetates, maleates, tartrates, fumarates, methanesulfonates, benzenesulfonates, tosylates, etc.). Furthermore, mention is also made of salts with basic or acidic amino acids (for example salts with arginine, aspartic acid, glutamic acid, etc.).

The cyclic polypeptide compound (I) of the present invention also includes possible conformational isomers and one or more pairs of stereoisomeric forms, such as geometric isomers and optical isomers, which may exist due to asymmetric carbon atoms.

Preferably, the cyclic polypeptide compound (I) is represented by the following formulae (II) to (VI):

(continue on the next page)

The most preferred cyclic polypeptide compounds are represented by formula (II).

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

As the stabilizer, there may be mentioned polysaccharides, disaccharides, sodium chloride and combinations thereof.

Examples of the polysaccharide are dextran, starch, cellulose and hyaluronic acid; examples of disaccharides are lactose, maltose and sucrose. The polysaccharide or disaccharide included in the pharmaceutical composition of the invention may be an alpha-monohydrate, an alpha-anhydride, a beta-anhydride or a combination thereof.

The amount of the stabilizing agent in the pharmaceutical composition of the present invention should be at least sufficient to stabilize the cyclic polypeptide compound (I) or a pharmaceutically acceptable salt thereof in the composition. For stabilizing the cyclic polypeptide compound (I), at least one part by weight of the stabilizing agent corresponding to one part by weight of the cyclic polypeptide compound (I) or a pharmaceutically acceptable salt thereof in the composition of the present invention is sufficient. The stabilizer may also be used as a carrier or excipient. Therefore, the amount of the stabilizer to be used does not have a specific upper limit, and may be determined in consideration of the weight or volume of the composition, etc., according to one unit dose of the compound. However, such an 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, relative to one part by weight of the cyclic polypeptide compound (I) or a pharmaceutically acceptable salt thereof, although the amount may vary depending on the kind and amount of the cyclic polypeptide compound (I) or a pharmaceutically acceptable salt thereof, a preparation form thereof, and/or the like. In particular, the disaccharide is more preferably used in an amount of 1 to 20 parts by weight, still more preferably 2 to 10 parts by weight, relative to one part by weight of the cyclic polypeptide compound (I) or a pharmaceutically acceptable salt thereof. In particular, the cyclic polypeptide compound (I) or a pharmaceutically acceptable salt thereof is used in an amount of more preferably 0.6 to 20 parts by weight, and still more preferably 0.8 to 10 parts by weight, relative to one part by weight of the cyclic polypeptide compound (I).

The pharmaceutical compositions of the present invention can be produced according to methods known in the art, using additives as necessary. Here, as a reference, there may be mentioned Basic characteristics on development of Pharmaceuticals XI 20 Production of Pharmaceuticals (volume 2) (edited by Kyobunn Tsuda and Hisashi Nogami, published by Chizyo Shoten). The lyophilized composition can be obtained by preparing an aqueous solution of the cyclic polypeptide compound (I) or a pharmaceutically acceptable salt thereof and the stabilizer, optionally adding a pH adjusting agent (citric anhydride, sodium hydroxide, etc.) as needed to achieve a pH of 4.0 to 7.5, preferably 4.5 to 7.0, and then lyophilizing the resulting solution in a vial according to a conventional method. Thus, the stable pharmaceutical composition in lyophilized form, when dissolved in pure water, preferably results in a solution having a pH of 4.0-7.5, more preferably a solution having a pH of 4.5-7.0. Preferably, the composition thus prepared in lyophilized form is sealed and stored protected from light. The lyophilized composition may be filled into each vial as a solution prior to lyophilization or as a lyophilized powder after lyophilization.

Since the stability of the cyclic polypeptide compound with respect to humidity is not satisfactory, it is essential that the lyophilized composition of the present invention contains 3.4% by weight or less of water, preferably 3.0%, more preferably 2.0% of water.

The stable pharmaceutical compositions in lyophilized form are typically dissolved as desired in isotonic sodium chloride solution and used as injection solutions. The pharmaceutical composition of the present invention can be used as an injectable preparation, which requires some mixing prior to use.

Best mode for carrying out the invention

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

Example 1

Compound (II)25g

Lactose 200g

Appropriate amount of anhydrous citric acid

Proper amount of sodium hydroxide

Lactose was dissolved in purified water (2000ml) with heating below 50 ℃. After cooling to below 20 ℃, compound (II) was added to the lactose solution, avoiding the formation of air bubbles under gentle stirring. After adding a 2% aqueous citric acid solution (9.5ml), a 0.4% aqueous sodium hydroxide solution (about 24ml) was added to the solution to adjust ph5.5, followed by dilution with pure water to give a given volume (2500 ml). The resulting solution was dispensed into 1,000 vials of l0ml volume, 2.5ml each. The solution in each vial was lyophilized by a lyophilizer (RL-603 BS manufactured by Kyowa Shinku co., Ltd) by a conventional method to obtain lyophilized compositions each containing 25mg of compound (II).

Example 2

Lyophilized compositions each containing 50mg of compound (II) were obtained in the same manner as in example 1 except that the amount of compound (II) was 50 g.

Example 3

Lyophilized compositions each containing 25mg of compound (II) were obtained in the same manner as in example 1 except that 150g of maltose was used instead of lactose.

Example 4

Lyophilized compositions each containing 50mg of compound (II) were obtained in the same manner as in example 1 except that compound (II) was used in an amount of 50g instead of 25g, and that lactose was replaced with 250g of sucrose.

Example 5

Lyophilized compositions each containing 25mg of compound (II) were obtained in the same manner as in example 1 except that 25g of sodium chloride was used in place of lactose.

Example 6

Lyophilized compositions each containing 10mg of compound (II) were obtained in the same manner as in example 1 except that the amount of compound (II) was 10g instead of 25g, and 100g of dextran was used instead of lactose.

Example 7

Lyophilized compositions each containing 25mg of compound (III) were obtained in the same manner as in example 1 except that compound (II) was replaced with 25g of compound (III) and lactose was replaced with 200g of maltose.

Example 8

Lyophilized compositions each containing 10mg of compound (IV) were obtained in the same manner as in example 1 except that compound (II) was replaced with 10g of compound (IV), and the amount of lactose was 100g instead of 200 g.

Example 9

Lyophilized compositions each containing 50mg of compound (V) were obtained in the same manner as in example 1 except that compound (II) was replaced with 50g of compound (V), and lactose was replaced with 50g of sodium chloride.

Example 10

Lyophilized compositions each containing 10mg of compound (VI) were obtained in the same manner as in example 1 except that compound (II) was replaced with 10g of compound (VI) and lactose was replaced with 100g of dextran.

Test example 1

Effect of stabilizers in Stable lyophilized compositions of Compound (II)

10mg of compound (II) and 100mg of lactose or 9mg of sodium chloride as stabilizer were completely dissolved in 1ml of water. The resulting solution was freeze-dried and kept at 70 ℃ in a glass vial. After 9 days, the resulting composition was tested for appearance, residual amount of compound (II), and other test contents. As a control, a solution of compound (II) without any stabilizer was used. The results are shown in Table 1.

TABLE 1

Stabilizer	Test items	0 hour	After 9 days
				Comparison: is free of	Appearance of the product	White block	Yellowish block
pH*	7.1	2.7
			Residual (%)		100.0	8.0
Water content (%)	1.3	-
			Lactose (100mg)		Appearance of the product	White block	White block
pH*	6.4	6.1
				Residual (%)	100.0	99.5
Water content (%)	1.0	-
				Sodium chloride (9mg)	Appearance of the product	White block	White block
pH*	6.7	6.3
			Residual (%)		100.0	75.9
Water content (%)	0.7	-

^*pH of the reconstituted composition solution in 1ml of water

Test example 2

A similar experiment was conducted in the same manner as in test example 1 except that 100mg of maltose, 50mg of sucrose or 50mg of glucose was used as a stabilizer. The results are shown in Table 2.

(continue on the next page)

TABLE 2

Stabilizer	Test items	0 hour	After 9 days
				Comparison: is free of	Appearance of the product	White block	White block
pH*	6.8	5.4
			Residual (%)		100.0	＜75.0
Water content (%)	3.3	-
			Maltose (100mg)		Appearance of the product	White block	White block
pH*	7.3	6.7
				Residual (%)	100.0	98.6
Water content (%)	0.9	-
				Sucrose (50mg)	Appearance of the product	White block	White solution
pH*	6.9	7.0
			Residual (%)		100.0	82.4
Water content (%)	1.1	-
			Glucose (50mg)		Appearance of the product	White solution	Brown solution
pH*	6.9	3.6
				Residual (%)	100	1.1
Water content (%)	4.3	-

^*pH of the reconstituted composition solution in 1ml of water

As is evident from tables 1 and 2, the lyophilized compositions of compound (II) and lactose, sodium chloride, maltose or sucrose are significantly stable compared to compositions without any mild stabilizer or other stabilizer.

Test example 3

Dependence of the stability of the lyophilized composition of Compound (II) on the amount of lactose added

An experiment was performed in the same manner as in test example 1 except that 20mg, 50mg, 100mg or 200mg of lactose was used as a stabilizer. Table 3 shows the test results by observing the appearance of the composition, the residual amount of compound (II), the appearance of the composition solution reconstituted in 1ml of water, and the like. Incidentally, it took 15 seconds to reconstitute the composition in 1ml of water.

TABLE 3

Lactose addition amount (mg)	Test items	0 hour	After 9 days at 70 DEG C	After 3 months at 40 ℃ and 75% humidity
					20	Appearance of the product	White block	Yellowish block	White block
Colour(s)*	Colorless and colorless	White colour	Colorless and colorless
				Transparency*		Is transparent	Is not transparent	Is transparent
pH*	6.09	3.03	6.57
				Residual (%)		100.0	88.09	100.0
Total impurities (%)	3.44	12.3	3.99
				Water content (%)		1.2	-	-
50	Appearance of the product	White block	White block						White block
				Colour(s)*	Colorless and colorless	Colorless and colorless	Colorless and colorless
	Transparency*	Is transparent	Is transparent					Is transparent
				pH*	6.57	5.56	6.26
	Residual (%)	100.0	96.7					99.8
				Total impurities (%)	3.32	7.37	4.21
	Water content (%)	0.5	-					-
				100	Appearance of the product	White block	White block		White block
Colour(s)*	Colorless and colorless	Colorless and colorless	Colorless and colorless
					Transparency*	Is transparent	Is transparent	Is transparent
pH*	6.58	6.08	5.80
					Residual (%)	100.0	96.7	99.6
Total impurities (%)	3.43	7.08	3.96
					Water content (%)	0.3	-	-
200	Appearance of the product	White block	White block						White block
				Colour(s)*	Colorless and colorless	Colorless and colorless	Colorless and colorless
	Transparency*	Is transparent	Is transparent					Is transparent
				pH*	6.78	5.70	5.36
	Residual (%)	100.0	96.1					99.6
				Total impurities (%)	3.40	7.30	4.35
	Water content (%)	0.3	-					-

^*Color, clarity and pH of the reconstituted composition solution in 1ml of water

As is evident from table 3, the lyophilized compositions of 10mg of compound (II) and varying amounts of lactose had no problem in their stability.

Test example 4

Stability in vials of a lyophilized composition of 200mg lactose and Compound (II)

An experiment was performed in the same manner as in test example 1 except that 12.5mg, 25mg, 50mg, 75mg or 100mg of compound (II) and 200mg of lactose were used. Table 4 shows the test results for the residual amount of compound (II) in the obtained composition, and the like. With respect to all of the compositions, the appearance was a white mass, the reconstitution dissolution time was 15 seconds, and the color and transparency of the reconstituted solution of the composition were colorless and transparent.

TABLE 4

Amount of Compound (II) added (mg)	Test items	0 hour	After 9 days at 70 DEG C	After 21 days at 60 DEG C	After 3 months at 40 ℃ and 75% humidity
						12.5	pH*	6.63	6.15	6.31	6.08
Residual (%)	100.0	98.1	97.5	99.6
					Total impurities (%)		2.24	3.95	3.75	2.71
Water content (%)	1.3	-	-
					25		pH*	6.37	6.07	6.11	6.14
Residual (%)	100.0	99.3	98.2	101.2
						Total impurities (%)	2.25	4.03	3.49	2.68
Water content (%)	1.1	-	-	-
						50	pH*	6.26	5.99	6.00	6.00
Residual (%)	100.0	97.9	97.3	100.5
					Total impurities (%)		2.25	3.95	3.68	2.74
Water content (%)	1.2	-	-	-
					75		pH*	6.13	5.95	5.96	6.04
Residual (%)	100.0	98.1	97.7	99.0
						Total impurities (%)	2.28	4.14	3.83	2.76
Water content (%)	0.9	-	-	-
						100	pH*	6.03	5.92	5.88	5.85
Residual (%)	100.0	97.8	96.7	99.5
					Total impurities (%)		2.46	4.15	3.92	2.79
Water content (%)	1.3	-	-	-

^*pH of the reconstituted composition solution in 5ml of purified water

As is evident in table 4, all lyophilized compositions were stable.

Test example 5

Stability test

The pharmaceutical compositions obtained in examples 1 and 2 were stored at room temperature. After 18 months, the residual ratio of compound (II) was 98% in all the compositions.

Test example 6

Dependence of the lyophilized composition of Compound (II) on the pH of a solution of said composition before lyophilization

10mg of compound (II), 100mg of lactose as stabilizer, were completely dissolved in 1ml of citrate-NaOH buffers with different pH values of 4.0-7.0. The resulting solutions with different pH values were freeze dried and kept in glass vials at 70 ℃. After 9 days, the resulting composition was tested for pH and residual amount of compound (II). The results are shown in Table 5.

TABLE 5

	pH of the composition solution before lyophilization	4.0	4.5	5.0	5.5	6.0	6.5	7.0
									0 hour	pH*	3.9	4.4	4.8	5.4	5.8	6.4	6.8
Water content (%)	0.2	0.2	0.3	0.3	0.3	0.4	0.3
								Residual (%)		100
9 days	pH*	4.0	4.5	4.9	5.4	5.8	6.4										6.8
								Residual (%)	94.4	95.9	97.4	98.5	97.7	96.9	95.8

^*pH of the reconstituted composition solution in 5ml of purified water

As is apparent from Table 5, the pharmaceutical compositions of the present invention are stable after lyophilization of a solution containing compound (II) at a pH of at least 4.0 to 7.0, preferably at a pH of 4.5 to 7.0.

Test example 7

Dependence of the lyophilized composition of Compound (II) on the Water content of the composition

10mg of Compound (II), 50mg of lactose as a stabilizer were completely dissolved in 1ml of water. The resulting solution was freeze dried and kept in glass vials at 70 ℃. After 9 days, the resulting composition was tested for pH, its water content and the residual amount of compound (II). The results are shown in Table 6.

TABLE 6

Water content at 0 hour		0.9	1.4	2.6	3.4	5.1
							0 hour	pH*	7.1
After 9 days	pH*	7.5	7.1	6.8	6.8	3.5
							Water content (%)	2.5	2.9	3.6	4.3	5.4
	Residual (%)	97.6	98.1	97.1	92.7	18.3

^*pH of the reconstituted composition solution in 1ml of water

As is evident from Table 6, the pharmaceutical compositions of the present invention are stable and contain about 3.5%, more preferably 3.4% by weight or less of water.

According to the present invention, there is provided a composition in lyophilized form in which the cyclic polypeptide compound (I) or a pharmaceutically acceptable salt thereof is stabilized with a stabilizing agent such as a polysaccharide, a disaccharide and sodium chloride.

The mechanism of stabilizing the cyclic polypeptide compound (I) or a pharmaceutically acceptable salt thereof with stabilizers such as polysaccharides, disaccharides and sodium chloride is not known, but may be: the stabilizer absorbs moisture in the lyophilized cake, and the stabilizer serves to uniformly disperse the compound or a pharmaceutically acceptable salt thereof in the composition.

The cyclic polypeptide compounds (I) have antifungal activity, in particular against:

acremonium (Acremonium);

absidia (Absidia) (e.g., Absidia corymbifera, etc.);

aspergillus (Aspergillus) (e.g., Aspergillus clavatus, Aspergillus flavus, Aspergillus fumigatus, Aspergillus nidulans, Aspergillus niger, Aspergillus terreus, Aspergillus versicolor, etc.);

blastomyces (Blastomyces) (e.g., Blastomyces dermatitidis (Blastomyces dermatitidis), etc.);

candida (Candida) (e.g., Candida albicans, Candida glabrata, Candida quaternary (Candida guilliermondii), Candida lactis (Candida kefyr), Candida krusei (Candida krusei), Candida parapsilosis (Candida parapsilosis), Candida stellatoides (Candida stellatoides), Candida tropicalis (Candida tropicalis), Candida utilis (Candida utilis), and the like);

cladosporium (Cladosporium) (e.g., Cladosporium trichoides, etc.);

coccidioidomycosides (e.g., coccoidoidioides immitis, etc.);

cryptococcus (Cryptococcus) (e.g., Cryptococcus neoformans (Cryptococcus of ormans), etc.);

cunninghamella (e.g., Cunninghamella elegans, etc.);

genus dermatophytes (Dermatophyte);

exophiala (Exophiala) (e.g., Exophiala dermatitidis (Exophialadaptitis), Exophiala spinofera, etc.);

epidermophyton (Epidermophyton) (e.g., Epidermophyton floccosum (Epidermophyton coccus), etc.);

genus chromoblastomyces (Fonsecaea) (e.g., chromoblastomyces eupatoria (fonsecaeapedresori), etc.);

fusarium (Fusarium) (e.g., Fusarium solani, etc.);

geotrichum (Geotrichum) (e.g., Geotrichum candidum);

histoplasma (Histoplasma) (e.g., Histoplasma capsulatum var. capsulatum), etc.);

malassezia (Malassezia) (e.g., Malassezia furfur (Malassezia furfur), etc.);

microsporum (Microsporum) (e.g., Microsporum canis, Microsporum gypseum, etc.);

mucor (Mucor);

coccidioidomycosis (Paracoccidioides) (e.g., Paracoccidioides brasiliensis (Paracoccidioides), etc.);

penicillium (Penicillium) (e.g., Penicillium marneffei, etc.);

phialophora (Phialophora);

pneumocystis (Pneumocystis) (e.g., Pneumocystis carinii (Pneumocystis carinii), etc.);

pseudomyceliophthora (pseudoallomycelia) (e.g., pseudomyceliophthora boydii) and the like);

rhizopus (Rhizopus) (e.g., Rhizopus microsporus var. Rhizopus, Rhizopus oryzae, etc.);

saccharomyces (Saccharomyces) (e.g., Saccharomyces cerevisiae), etc.);

scopulariopsis (Scopulariopsis);

sporothrix (e.g., Sporothrix schenchi et al);

trichophyton (e.g., Trichophyton mentagrophytes, Trichophyton rubrum, etc.);

trichosporon (Trichosporon) (e.g., Trichosporon asahii, Trichosporon cutaneum), etc.).

It is known that the above fungi cause various infectious diseases in skin, hair, nails, oral mucosa, gastrointestinal tract, bronchus, lung, endocardium, brain, meninges, urinary organs, vaginal promotion, oral cavity, eye, whole body, kidney, bronchus, heart, external auditory meatus, bone, nasal cavity, paranasal cavity, spleen, liver, subcutaneous tissue, lymphatic vessel, gastrointestinal tract, joint, muscle, tendon, lung interstitial plasma cells, and the like.

Thus, the cyclic polypeptide compound (I) of the present invention is useful for the prevention and treatment of various infectious diseases such as dermatophytosis (e.g., trichophytosis, etc.), pityriasis versicolor, candidiasis, cryptococcosis, geotrichosis, trichosporidiosis, aspergillosis, penicilliosis, fusarium, zygomycosis, sporotrichosis, chromomycosis, coccidioidomycosis, histoplasmosis, blastomycosis, coccidioidomycosis, pseudomycosis, mycetoma, fungal keratitis, otomycosis, pneumocystis, etc.

The commercial package comprises the cyclic polypeptide compound (I) of the composition of the invention and the written matter which states that the pharmaceutical composition can or should be used for the prevention or treatment of infectious diseases.

Claims (6)

1. A stable pharmaceutical composition in lyophilized form comprising a water content of 3.0% by weight or less, said composition comprising as an active ingredient a cyclic polypeptide compound of general formula (I):

or a pharmaceutically acceptable salt thereof, and maltose as a stabilizer.

2. A pharmaceutical composition according to claim 1, which is prepared by the steps of:

■ dissolving the cyclic polypeptide compound (I) or a pharmaceutically acceptable salt thereof, the stabilizer and optionally a pH adjuster in pure water, and

■ the solution is lyophilized.

3. The pharmaceutical composition according to claim 1, which, when dissolved in pure water, results in a solution having a pH of 4.0 to 7.5.

4. The pharmaceutical composition according to claim 1, additionally comprising a pH adjusting agent.

5. An injectable formulation prepared by dissolving the pharmaceutical composition of claim 1 in an isotonic sodium chloride solution.

6. A commercial package comprising a pharmaceutical composition according to any one of claims 1 to 4 and written matter associated therewith, wherein said written matter states that said pharmaceutical composition can or should be used for the prevention or treatment of an infectious disease.