JPH0687881A - Rokitamycin monohydrate crystal and its production - Google Patents

Abstract

PURPOSE:To provide a new compound usable without masking bitter taste because of little bitterness compared with rokitamycin, capable of preparing a syrup preparation having excellent quality with simple process and exhibiting excellent wetting resistance to dispense with the surface treatment. CONSTITUTION:Rokitamycin monohydrate of formula. The compound can be produced by crystallizing rokitamycin(RKM) in an acidic aqueous solution (preferably an aqueous solution of pH1-4 prepared by using acetic acid, propionic acid, etc.). Concretely, RKM anhydride and/or methanol-containing RKM crystal are dissolved in an acidic aqueous solution at a concentration of 20-300mg/ml under cooling and stirring at 0-10 deg.C, the pH is adjusted to 4.4-5 with dilute alkaline water, the liquid temperature is slowly raised to the room temperature and the solution is left standing for 2-72hr to form the objective crystal. The RKM to be used as a raw material is preferably methanol-containing RKM crystal to get high-purity RKM monohydrate.

Description

Detailed Description of the Invention

[0001]

The present invention relates to the formula (1)

[0002]

[Chemical 3]

The present invention relates to a rokitamycin monohydrate crystal represented by: and a method for producing a rokitamycin monohydrate crystal represented by the formula (1), which comprises crystallizing rokitamycin in an acidic aqueous solution.

[0004]

2. Description of the Related Art Rokitamycin (hereinafter abbreviated as RKM) has the formula (2)

[0005]

[Chemical 4]

A compound represented by the following formula, which is a kitasamycin producing bacterium Streptomyces kitasatoensis (Str
It is a semi-synthetic macrolide antibiotic obtained by selectively propionylating the 3'-tertiary hydroxyl group from leucomycin A ₅ produced by a mutant strain belonging to Eptomyces kitasatoensis. Is a patent (Japanese Patent Publication No. 59-46520)
Publication) and literature (J. Antibiotics, 3
4,1001 (1981)). Its antibacterial activity is stronger than the existing 16-membered ring macrolides kitasamycin, josamycin, midecamycin, and myocamycin, and is comparable to 14-membered ring erythromycin (Id.). In addition, RKM is a drug showing an excellent antibacterial action against resistant bacteria in which existing macrolide antibiotics are completely ineffective, and for example, tablets for oral administration containing glycine and citric acid (US Pat. No. 4,716,153). It is highly safe and is widely used today.

Conventionally, RKM has been industrially synthesized by using leucomycin A ₅ as a raw material, the obtained crude product is purified by column chromatography, the obtained eluate is concentrated, and the concentrate is dropped into water. The precipitated precipitate was collected by filtration,
This was dried and made into a product. The bulk powder crystallized in this manner is amorphous and becomes RKM in an anhydrous state (hereinafter abbreviated as RKM anhydrous), and the X-ray diffraction spectrum of this RKM anhydrous is shown in FIG. It was also clear that it was amorphous because it showed a broad spectrum having peaks at diffraction angles (2θ) of 12 and 19 °.

[0008]

Further, this amorphous R
KM anhydrous has a strong bitterness, and when formulated as a syrup, the bitterness was improved by pretreating RKM by a spray drying method using a polymer film as a coating agent. Furthermore, from the research on the physicochemical properties of RKM anhydride, RKM anhydride itself is
Since it is highly fat-soluble and hard to get wet with water, 4% of sucrose fatty acid ester (hereinafter abbreviated as DK ester), which is usually used for oral preparations with high safety, is used for tablets, and surface treatment is performed. The preparation was made to improve dispersibility in gastric juice and to dissolve rapidly by applying the method (Japanese Patent Publication No. 2-59129, Pharmaceutical Science, 48, 147 (19).
88)). As described above, RKM anhydrous had a strong bitterness and a property that it was difficult to wet with water. Also, R
In the production of KM bulk powder, it was difficult to further improve the quality by the purification method using only the above column chromatography step.

Therefore, when the purification method was investigated for the purpose of improving the quality of RKM, it was possible to obtain a novel RKM crystal containing methanol by crystallizing RKM anhydrous using methanol. The X-ray diffraction spectrum of this RKM crystal containing methanol is shown in FIG. This is 7.8,
8.3, 10.3, 10.7, 12.1, 12.6, 1
4.2, 15.8, 16.6, 18.2, 20.3, 2
1.7, 22.1, 23.2, 24.0 and 25.5 °
Since it showed a sharp diffraction spectrum at the diffraction angle (2θ) of 1, it was clear that it was crystalline. However, this product also had a bitter taste.

[0010]

The inventors of the present invention have conducted extensive studies to solve the above problems, and as a result, there have been no reports of monohydrates of macrolide antibiotics. RKM crystals containing anhydride or methanol are dissolved in an organic acid such as acetic acid, propionic acid, tartaric acid, and citric acid, or an aqueous solution of an inorganic acid such as phosphoric acid, hydrochloric acid, and sulfuric acid to be crystallized, thereby providing a simple and high yield It was found that a completely new RKM monohydrate crystal was obtained in. Moreover, surprisingly, this RKM monohydrate was obtained as a crystalline substance in which the bitterness of the RKM bulk powder was hardly felt, and further, it was highly pure and had high thermal stability and improved water wetting. The inventors have found that they have properties and completed the present invention.

The present invention has been completed on the basis of the above findings, and has the formula (1)

[0012]

[Chemical 5]

RKM monohydrate crystal represented by:
A method for producing an RKM monohydrate crystal represented by the formula (1), characterized in that RKM is crystallized in an acidic aqueous solution. The RKM as a raw material used in the present invention is not limited as long as it contains the crude or appropriately purified RKM obtained by the above-mentioned synthetic procedure, and for example, RKM anhydrous or methanol. The contained crystals can be easily used, and in particular, the RKM methanol-containing crystals can be used to obtain highly pure RKM monohydrate crystals.

To obtain the RKM monohydrate crystals of the present invention, first, an acidic aqueous solution, for example, an organic acid such as acetic acid, propionic acid, tartaric acid or citric acid, or an inorganic acid such as phosphoric acid, hydrochloric acid or sulfuric acid is used. PH of acid or mixed acid
Prepare an aqueous solution of 1 to 4. Preferred are organic acids such as acetic acid and propionic acid, or inorganic acids such as phosphoric acid. Then, the acidic aqueous solution thus prepared is cooled to 0 ° C. to 10 ° C. and stirred, and the RKM anhydrous and / or RKM crystals containing methanol are dissolved at a concentration of 20 to 300 mg / ml. Alkaline water obtained by diluting sodium hydroxide, sodium carbonate, sodium hydrogencarbonate, potassium hydroxide, potassium carbonate, potassium hydrogencarbonate, ammonium carbonate, ammonium water, etc. to a concentration of 5 to 20% in order to crystallize after completely dissolving. PH is adjusted to, for example, about 4 to 5.5, preferably about 4.4 to 5.0, and the solution is adjusted to 20 to 3
Gradually raise the temperature to 0 ° C. and let stand for about 2 to 72 hours to form crystals. The grown crystals are collected by filtration, washed with water and dried under reduced pressure to obtain the desired RKM monohydrate. Crystals can be obtained.

The RKM of the present invention thus obtained
The X-ray diffraction spectrum of the monohydrate crystal (hereinafter abbreviated as RKM hydrate) is shown in FIG. This is 5.2, 6.8, 8.
3, 10.3, 11.2, 12.3, 13.3, 13.
8, 14.7, 15.1, 15.8, 16.4, 17.
1, 18.1, 19.1, 19.5, 20.4, 21.
5, 22.0, 22.7, 23.8, 24.8, 25.
It is clear that it is crystalline because it shows a sharp diffraction spectrum at diffraction angles (2θ) of 1, 26.6 and 28.6 °. The RKM hydrate of the present invention is clearly different from the above RKM anhydrous and methanol-containing crystals in the 2θ value of the diffraction spectrum. Further, the RKM hydrate of the present invention has 1 water molecule per 1 molecule of RKM from the results of instrumental analysis such as elemental analysis, moisture measurement (Karl Fischer method) and thermogravimetric analysis.
It was identified that they were individually hydrated, and it was found to be a new crystalline form.

The antibacterial activity of the novel RKM hydrate of the present invention was the same as that of conventional RKM anhydrous. Furthermore, the RKM hydrate of the present invention did not cause any acute toxicity to mice (5 mice per group) by intraperitoneal administration at 2000 mg / kg. The novel RKM hydrate thus obtained has almost no bitter taste and is well wetted with water, and can be used predominantly in formulations for oral administration, particularly dry syrups and tablets. It was Furthermore, as a pharmaceutical advantage, the RK of the present invention
It has become possible to develop new drug formulations such as lozenges, granules, eye ointments, and ointments using M hydrate, which have been difficult to achieve in the past, and have extremely superior specificity from a pharmaceutical viewpoint. I have.

Further in the formulation, the RKM hydrate is 1
It may be used to administer 100 to 600 mg 1 to 3 times a day. In addition to these, RKM hydrate has a specific volume that is about 30% smaller than conventional RKM anhydrous, and has high hardness because it is a crystal, which enables downsizing and high hardness of tablets, and crushing. It has peculiarities such as little electrostatic charge at the time. Therefore, the RK of the present invention
M hydrate is a crystal having the above-mentioned excellent properties in formulation, and can be formed into an appropriate formulation by a formulation of a formulation in a conventional method.

[0018]

EXAMPLES Next, the present invention will be specifically described with reference to the following Reference Examples and Examples, but the present invention is not limited thereto.

[0019]

Reference Example 1 RKM Synthesis Method The synthesis method of RKM is a patent (Japanese Patent Publication No. 59-46520, Japanese Patent Publication No. 63-5037) or a document (JA.
ntibiotics, 34, 1001 (1981))
That is, it was synthesized according to the following steps using leucomycin A ₅ as a starting material, and isolated and purified as RKM anhydrous. (1) Protection reaction of 2'-hydroxyl group 50 g of leucomycin A ₅ bulk powder isolated from the culture solution was dissolved in 250 ml of anhydrous 1,2-dichloroethane, and 21 ml of acetic anhydride was added dropwise under stirring at room temperature to give 0.5 Reacted for hours. Next, add 1 L of ice-cold water and pour with 7% ammonia water.
After adjusting to H10, the mixture was stirred for about 1 hour, and the 1,2-dichloroethane phase was separated. After drying then the resulting 1,2-dichloroethane phase with anhydrous magnesium sulfate 10 g, concentrated to dryness under reduced pressure to 2'-O- acetyl leucomycin A ₅ 50
g was obtained. (2) Protection reaction of 3,9-hydroxyl group 50 g of the above 2'-O-acetylleucomycin A ₅ was dissolved in 250 ml of anhydrous 1,2-dichloroethane, and then 50 g of tribenzylamine was added. 33 ml of trimethylchlorosilane was added dropwise with stirring and cooling, and the mixture was reacted for 15 hours. Next, the reaction solution was dropped into 1 L of water prepared in advance in another container, and the pH was adjusted to 9.5 with 7% ammonia water,
The 1,2-dichloroethane phase was separated. Then, the 1,2-dichloroethane phase obtained was mixed with anhydrous magnesium sulfate 10
After drying with g, it was concentrated to dryness under reduced pressure and 2'-O-acetyl-
3,9-di-O-trimethylsilylleucomycin A ₅
63 g are obtained. (3) Propionylation Reaction of Tertiary Hydroxyl at 3 "Position 63 g of the above 2'-O-acetyl 3,9-di-O-trimethylsilylleucomycin A ₅ was dissolved in 250 ml of anhydrous 1,2-dichloroethane, and then tribenzdiene. After adding 150 g of ruamine, 50 ml of propionyl chloride was added dropwise under ice cooling and then refluxed for about 20 hours at 75 ° C. After completion of the reaction, the mixture was cooled to room temperature and then prepared in another container in advance. 2-dichloroethane 250m
1 and 1 L of water were added dropwise to the mixture, and p
After adjusting to H9.5, the 1,2-dichloroethane phase was separated. Then, the obtained 1,2-dichloroethane phase was dried over 10 g of anhydrous magnesium sulfate and then concentrated to dryness under reduced pressure.
2'-O-acetyl-17,18-enol-18,
3 "- di -O- propionyl-3,9-di -O- trimethylsilyl leucomycin A ₅ and 2'-O- acetyl -3" -O- propionyl-3,9-di -O- trimethylsilyl leucomycin A ₅ 68 g of a mixture of (4) Desorption reaction of protecting group 68 g of the above mixture was dissolved in a mixed solvent of 2.5 L of methanol and 275 ml of 8% calcium carbonate, and reacted at room temperature for 1 hour. The reaction solution was adjusted to pH 7.5 with acetic acid,
Then, the mixture was heated under reflux at 63 ° C. for 20 hours and then cooled to 5 ° C. or lower, and the tribenzylamine crystals formed in the reaction solution were separated by filtration. The crystals of tribenzylamine were washed with 220 ml of 90% methanol cooled to 5 ° C. or lower, the filtrate and the washing solution were combined, and methanol was completely distilled off under reduced pressure. The residue was then dissolved in 300 ml of benzene, partitioned and the remaining aqueous phase separated off. Benzene phase is water 30
After washing with 0 ml, the mixture was concentrated under reduced pressure to about 100 ml. (5) Isolation and Purification Silica gel column (30 mmφ) packed with benzene
Approximately 100 ml of the above benzene concentrate was adsorbed on (X600 mmL) and then developed with benzene: ethyl acetate: methanol (36: 4: 1, v / v). The eluate was sampled over time, analyzed by high performance liquid chromatography (hereinafter abbreviated as HPLC), and about 650 ml of fractions having an RKM content of 90% or more were collected and concentrated to dryness under reduced pressure. The concentrated residue was dissolved in 60 ml of ethanol, and the ethanol solution was added dropwise to 800 ml of water to deposit RKM as a white precipitate. This precipitate was collected by filtration, washed with water, and dried under reduced pressure to obtain 35 g of RKM.

The physicochemical properties of the RKM thus obtained are described in the patent (Japanese Patent Publication No. 59-46520) and the literature (J. Antibiotics, 34, 1001 (1).
981)) was in agreement with known results. Further, since the X-ray diffraction spectrum shows the same X-ray diffraction pattern as that shown in FIG. 1, it is clear that it is amorphous, and further, elemental analysis, water content measurement (Karl Fischer method) and thermal analysis. From the results of the gravimetric analysis, this RKM was identified to be anhydrous.

[0021]

Reference Example 2 Method for Producing RKM Methanol-Containing Crystals 35 g of the RKM anhydride obtained in Reference Example 1 above was dissolved in 120 ml of methanol under stirring at 40 ° C. (RKM concentration of about 300 mg / ml). Distilled water was added to this solution under stirring at 40 ° C. until it became cloudy, then cooled to 10 ° C. or lower, and stirred for further 20 hours to form crystals. The crystals were collected by filtration, washed with water, and washed with RKM. Crystals containing methanol (see Figure 2)
25 g (wet weight) was obtained.

[0022]

Example 1 Method for producing RKM hydrate 25 g of RKM methanol-containing crystals obtained in Reference Example 2
(Wet weight) 100m of 3% acetic acid water cooled to 10 ° C
It was dissolved in 1. Then pH with 10% sodium hydroxide
After adjusting the temperature to 5 and leaving it at room temperature (about 26 ° C.) for 24 hours, the resulting crystals are collected by filtration, washed with water, and dried under reduced pressure to obtain RK.
8 g of M hydrate was obtained.

When this crystal was analyzed by the X-ray diffraction method (X-ray diffractometer manufactured by Rigaku Denki Co., Ltd.), the pattern shown in FIG. 3 was shown. This was clearly different from the industrially produced RKM anhydride (see FIG. 1). Elemental analysis value (empirical formula;
C ₄₂ H ₆₉ NO ₁₅ · H ₂ O) is shown in Table 1.

[0024]

[Table 1]

Moisture (Karl Fischer moisture meter manufactured by Kyoto Electronics Manufacturing Co., Ltd.); about 2.1% (theoretical value; 2.13%). Melting point: around 112 ° C. Thermogravimetric analysis (Thermal analyzer manufactured by Mac Science Co., Ltd.); The weight change was about 2% near the melting point.

From the above instrumental analysis values,
Further, it was identified that this crystal contained one molecule of water per one molecule of RKM, because the weight loss was not observed even after drying. In addition, mass spectrometry (MASS) spectrum,
^The physicochemical properties obtained from instrumental analysis results such as ¹ H- and ¹³ C-nuclear magnetic resonance (NMR) spectra, infrared absorption (IR) spectra, ultraviolet absorption (UV) spectra, and optical rotation ([α] _D ). Patent (Japanese Patent Publication No. 59-46520)
Publication) and literature (J. Antibiotics, 3
4,1001 (1981)) in agreement with the known result.

Further, RKM using a developing solvent of chloroform: methanol: glacial acetic acid: water (80: 7: 7: 1).
The result of thin layer chromatography of the hydrate was Rf value 0.3.
6 (sulfuric acid coloration, ultraviolet absorption, etc.) showed a single spot, which was in agreement with the Rf value of RKM. And its biological activity is S. aureus ATCC 6538P, S. p
yogenes N.Y. Y. 5, M.I. luteus AT
It showed the same antibacterial activity as conventional RKM anhydrous against assayed bacteria such as CC9341.

[0028]

Example 2 Method for Producing RKM Hydrate 500 g (wet weight) of RKM methanol-containing crystals obtained in the same manner as in Reference Example 2 above was dissolved in 2 L of 3% acetic acid water cooled to 10 ° C. After adjusting the pH to 5 with 10% aqueous sodium hydroxide, the RKM hydrate 5 obtained in Example 1 was used.
g, and allowed to stand at room temperature (about 26 ° C) for 24 hours, then the resulting crystals were collected by filtration, washed with water, and dried under reduced pressure to obtain RKM.
About 250 g of a hydrate (which showed the same X-ray diffraction pattern as shown in FIG. 3 and was the same as the crystal of Example 1) was obtained.

[0029]

Example 3 Method for Producing RKM Hydrate First, 6 ml of propion was dissolved in 100 ml of water to prepare 10
After dissolving 35 g (wet weight) of the RKM methanol-containing crystals obtained in the same manner as in Reference Example 2 above in acidic water cooled to 0 ° C., the pH was adjusted to 5 with 10% aqueous sodium hydroxide, and then obtained in Example 1. RKM hydrate crystals (1 g) was added, and the mixture was allowed to stand at room temperature (about 26 ° C.) for 24 hours, then the resulting crystals were collected by filtration, washed with water, and dried under reduced pressure to obtain RKM hydrate (same as shown in FIG. An X-ray diffraction pattern was shown, which was the same as the crystal of Example 1).

[0030]

Example 4 Method for Producing RKM Hydrate 35 g of RKM methanol-containing crystals (wet weight) obtained in the same manner as in Reference Example 2 above was dissolved in 100 ml of 3% phosphoric acid water cooled to 10 ° C. and then 10%. The pH was adjusted to 5 with aqueous sodium hydroxide, 1 g of the RKM hydrate crystal obtained in Example 1 was added, and the mixture was allowed to stand at room temperature (about 26 ° C.) for 24 hours, and the resulting crystal was collected by filtration and washed with water. After drying under reduced pressure, 10 g of RKM hydrate (which had the same X-ray diffraction pattern as shown in FIG. 3 and was the same as the crystal of Example 1) was obtained.

[0031]

Example 5 Method for Producing RKM Hydrate 35 g of RKM anhydride obtained in Reference Example 1 above was dissolved in 150 ml of 3% acetic acid water cooled to 10 ° C. Then 10
After adjusting the pH to 5 with aqueous sodium hydroxide solution, 1 g of the RKM hydrate obtained in Example 1 was added, and the mixture was allowed to stand at room temperature (about 26 ° C) for 2 hours.
After standing for 4 hours, the resulting crystals were collected by filtration, washed with water, and dried under reduced pressure to obtain RKM hydrate (the same X-ray diffraction pattern as shown in FIG. 3 was obtained, which was the same as the crystal of Example 1). Was)
16 g were obtained.

[0032]

EXPERIMENTAL EXAMPLE Next, a comparison experiment between the RKM hydrate of the present invention and the conventional industrially produced amorphous RKM anhydride will be described.

[0033]

Experimental Example 1 Stability test RKM hydrate and RKM obtained in the same manner as in Example 1
Each of the anhydrides was stored under severe conditions of 80 ° C. for 4 weeks, sampled daily, and the absolute content of RKM was determined by moisture measurement (Karl Fischer method) and HPLC analysis. The sample for HPLC measurement is about 5 mg of each RKM powder.
After weighing, it was dissolved in 5 ml of acetonitrile: 0.1% acetic acid water (1: 1) solution and adjusted. The result is shown in Figure 4.
(In the figure,-○-indicates RKM hydrate,-□-indicates RKM anhydrous). The RKM hydrate had better thermal stability than the anhydride.

Analysis conditions; column: stainless steel, 4 mmφ × 150 mm, Unis
il Q, C18, 5 μm Phase: 0.2 M ammonium acetate solution: Methanol: Acetonitrile = (31.5: 62.0: 6.5) Detector: Spectrophotometer (measurement wavelength: 232 nm) Column temperature: 40 ° C. Flow rate: 0.8 ml / min.

[0035]

[Experimental Example 2] Sensory test A syrup was prepared from RKM hydrate and anhydrate under the following formulation conditions. With respect to these syrups, a sensory test was conducted by 10 panelists (adults) selected. The results are shown in Table 2 (in the table, A: no bitterness is felt, B: neither can be said, C: bitter).

Formulation 1: RKM hydrate 2 g, glycine 0.
8 g and sucrose 12 g were mixed in a mortar and suspended in 80 ml of distilled water. Formulation 2: RKM anhydrous 2 g, glycine 0.8 g, white sugar 1
2 g were mixed in a mortar and suspended in 80 ml of distilled water. Formulation 3: RKM anhydrous 400 mg, glycine 0.8 g,
12 g of sucrose was mixed in a mortar and suspended in 80 ml of distilled water.

Formulation 4: RKM anhydrous 20 mg, glycine 0.8 g, and sucrose 12 g are mixed in a mortar, and distilled water 80 ml
Suspended in.

[0038]

[Table 2]

In the case of RKM hydrate, almost no bitterness was perceived. On the other hand, when RKM anhydrous was used as a microcapsule by a spray drying method together with a polymer film or the like to mask bitterness, RKM hydrate was used as it was.
Anhydrous had a bitterness sensed even with 1/100 of the amount of RKM hydrate, and the formulation using RKM hydrate had significantly improved bitterness.

[0040]

[Experimental Example 3] Dissolution (water wetting) test The dissolution test was performed using a powder obtained by mixing RKM hydrate and anhydrate in a mortar according to the formulation of the basic tablet composition.
Regarding the prescription of RKM hydrate, the prescription 5 is given below.
The anhydrous formulations are shown below as formulation 6 and were used in the following tests. The conditions for the dissolution test are
Acid-free gastric juice model 160 ml (physiological saline 40 ml + distilled water 120 ml) was kept at 37 ° C. in a 200 ml beaker, and a single clinical dose of the drug (containing 200 mg of RKM) was added under stirring with a stirrer (about 200 rpm). The eluate was sampled and filtered, and the drug concentration was measured by HPLC under the same conditions as above. As a control, an elution test was carried out using a powder (formulation 7) in which RKM anhydride was surface-treated with DK ester.

The results are shown in FIG. 5 (in the case of prescription 5) and FIG.
(For prescription 6) and FIG. 7 (for prescription 7) are shown. In order to estimate the bioavailability of a preparation (hereinafter abbreviated as BA), a dissolution test is generally performed, and this dissolution test method is a method known in the literature (Japanese Patent Publication No. 2-59129, Pharmacology. 48, No. 2, 147, 1988) was used.

Formulation 5: RKM Hydrate 200 mg Glycine 340 mg Citric Acid 70 mg Formulation 6: RKM Anhydrous 200 mg Glycine 340 mg Citric Acid 70 mg Formulation 7: DK Ester Surface Treated RKM Anhydrate (10 parts of RKM Anhydrate are placed in a mortar, A suspension of 0.4 parts by weight of sucrose fatty acid ester F-160 in 10 parts by weight of water was sufficiently moistened while mixing, and then dried under reduced pressure.) 210 mg Glycine 340 mg Citric acid 70 mg As is clear from FIG. 5, FIG. 6 and FIG. 7, RKM hydrate showed the same excellent dissolution curve as that of the formulation of the formulation 7 included as a control without any DK ester surface treatment as a pretreatment. However, in the case of anhydrous RKM, a slow elution rate was observed because it was difficult to wet with water. From these results, the RKM hydrate was a crystal having excellent wettability and having an excellent property of being rapidly dispersed in the dissolution test solution.

[0043]

[Experimental Example 4] Bulk density (specific volume) test A graduated test tube (10 ml) was filled with 2 g of each of RKM hydrate and anhydrous bulk powder, and tapping was performed from a height of 4 to 5 cm. The volume for was measured. The results are shown in Table 3. RKM hydrate had a specific volume of about 30% smaller than that of the anhydrous product, and it was possible to reduce the size of tablets and the like, and to make a formulation that is easy to drink.

[0044]

[Table 3]

[0045]

From the above results, the present invention provides a novel RKM hydrate having significantly improved bitterness than RKM. When this is used, RKM can be formulated as a syrup formulation. It is not necessary to microencapsulate RKM by the conventional spray-drying method using a polymer film as a coating agent, and it is possible to formulate a syrup agent of the same or better quality than conventional products without processing the bulk powder. It is possible, and since RKM hydrate is very excellent in water wetting, there is no need for conventional surface treatment with DK ester when formulated as tablets, and the dissolution test results show that
B. A. Also in the above, it is possible to formulate tablets of the same quality as conventional products. As described above, the RKM hydrate obtained by the present invention is a crystal having the above-mentioned extremely excellent properties even in formulation.

Further, the RKM hydrate of the present invention obtained by the above-mentioned production method has high purity, high yield and high thermal stability,
It is also possible to provide a separation and purification method capable of producing a single product of an active ingredient which meets the purpose or a product having a fixed component ratio on an industrial scale.

[Brief description of drawings]

FIG. 1 shows an X-ray diffraction spectrum of RKM anhydrous.

FIG. 2 shows an X-ray diffraction spectrum of a methanol-containing RKM crystal.

FIG. 3 shows an X-ray diffraction spectrum of RKM hydrate obtained according to the present invention (Example 1).

FIG. 4 shows thermal stability curves of RKM hydrate and RKM anhydrous obtained at 80 ° C. according to the present invention (Example 1).

FIG. 5 shows the dissolution curve of the acid-free gastric juice model in Formulation 5.

FIG. 6 shows the dissolution curve of the acid-free gastric juice model in Formulation 6.

FIG. 7 shows the dissolution curve of the acid-free gastric juice model in Formulation 7.

─────────────────────────────────────────────────── ───

[Procedure amendment]

[Submission date] September 27, 1993

[Procedure Amendment 1]

[Document name to be amended] Statement

[Correction target item name] 0006

[Correction method] Change

[Correction content]

A compound represented by the following formula, which is a kitasamycin producing bacterium Streptomyces kitasatoensis (Str
It is a semi-synthetic macrolide antibiotic obtained by selectively propionylating the 3'-tertiary hydroxyl group produced by reducing the amount of leucomycin A ₅ produced by a mutant strain belonging to Eptomyces kitasatoensis. Is a patent (Japanese Patent Publication No. 59-46520)
Publication) and literature (J. Antibiotics, 3
4,1001 (1981)). Its antibacterial activity is stronger than the existing 16-membered ring macrolides kitasamycin, josamycin, midecamycin, and myocamycin, and is comparable to 14-membered ring erythromycin (Id.). Further, RKM is a drug showing an excellent antibacterial action against resistant bacteria in which existing macrolide antibiotics are completely ineffective, and for example, a preparation for oral administration containing glycine and citric acid (US Pat. No. 4,716,153). It is highly safe and is widely used today. These 16-membered rings
Or 14-membered ring macrolide antibiotics have strong bitterness
The drawbacks it had were known. 14-member ring macrora
Of erythromycin and its derivatives
Monohydrate and dihydrate crystals (Journal of Ph
armature Science, 67,
(8), pp1087 (Aug. 1978), US Patent
No. 2864817, JP-A-64-38096
(Gazette) is known, but this erythromycin and its
Hydrate crystals of the derivative are obtained for the purpose of improving its solubility.
It has been done. However, these compounds
Orally, like any other macrolide antibiotic
It had a strong bitterness and no improvement in bitterness.
It was not done.

[Procedure Amendment 2]

[Document name to be amended] Statement

[Correction target item name] 0010

[Correction method] Change

[Correction content]

[0010]

Therefore, as a result of earnest research to solve the above-mentioned problem of bitterness , the present inventor reported a monohydrate of 16-membered ring macrolide antibiotics. Whereas, RKM anhydride or methanol-containing RKM crystals are crystallized by dissolving them in an aqueous solution of an organic acid such as acetic acid, propionic acid, tartaric acid or citric acid, or an inorganic acid such as phosphoric acid, hydrochloric acid or sulfuric acid. It was found that a completely new RKM monohydrate crystal can be obtained simply and in high yield. Moreover, surprisingly, this RKM monohydrate was obtained as a crystalline substance in which the bitterness of the RKM bulk powder was hardly felt, and further, it was highly pure and had high thermal stability and improved water wetting. The inventors have found that they have properties and completed the present invention.

[Procedure 3]

[Document name to be amended] Statement

[Correction target item name] 0014

[Correction method] Change

[Correction content]

To obtain the RKM monohydrate crystals of the present invention, first, an acidic aqueous solution, for example, an organic acid such as acetic acid, propionic acid, tartaric acid or citric acid, or an inorganic acid such as phosphoric acid, hydrochloric acid or sulfuric acid is used. PH of acid or mixed acid
Adjust to an aqueous solution of 1-4. Preferred are organic acids such as acetic acid and propionic acid, or inorganic acids such as phosphoric acid. Next, the thus prepared acidic aqueous solution is cooled to 0 ° C. to 10 ° C. and stirred, and the RKM anhydrous and / or methanol-containing RKM crystals are dissolved at a concentration of 20 to 300 mg / ml. After being completely dissolved, in order to crystallize, sodium hydroxide, sodium carbonate, sodium hydrogencarbonate, potassium hydroxide, potassium carbonate, potassium hydrogencarbonate, ammonium carbonate, ammonium water, etc., are diluted to a concentration of 5 to 20% in alcal water. Then, for example, the pH is adjusted to around 4 to 6.5 , preferably around 4.4 to 5.0, and the solution is adjusted to 20 to 3
Gradually raise the temperature to 0 ° C. and let stand for about 2 to 72 hours to form crystals. The grown crystals are collected by filtration, washed with water and dried under reduced pressure to obtain the desired RKM monohydrate. Crystals can be obtained.

[Procedure amendment 4]

[Document name to be amended] Statement

[Name of item to be corrected] 0023

[Correction method] Change

[Correction content]

When this crystal was analyzed by the X-ray diffraction method (X-ray diffractometer manufactured by Rigaku Denki Co., Ltd.) , the pattern of FIG. 3 was shown. This is the RKM anhydride (Fig.
See)). Elemental analysis value (empirical formula; C
₄₂ H ₆₉ NO ₁₅ · H ₂ O) is shown in Table 1.

[Procedure Amendment 5]

[Document name to be amended] Statement

[Name of item to be corrected] 0025

[Correction method] Change

[Correction content]

Water content (Karl Fischer water content meter, manufactured by Kyoto Electronics Manufacturing Co., Ltd.); about 2.1% (theoretical value as monohydrate ; 2.
13%). Melting point: around 112 ° C. Thermogravimetric analysis (Thermal analyzer manufactured by Mac Science Co., Ltd.); The weight change was about 2% near the melting point. Differential scanning calorimetry (Thermal analyzer manufactured by Mac Science);
An endothermic peak of about 20 cal / g was observed near the melting point.
It was

[Procedure correction 6]

[Document name to be amended] Statement

[Correction target item name] 0039

[Correction method] Change

[Correction content]

In the case of RKM hydrate, almost no bitterness was perceived. On the other hand, when RKM anhydrous was used as a microcapsule by a spray drying method together with a polymer film or the like to mask bitterness, RKM hydrate was used as it was.
Anhydrous had a bitterness sensed even with 1/100 of the amount of RKM hydrate, and the formulation using RKM hydrate had significantly improved bitterness. Reference example 3 RKM hydrate 100 g titer and Japanese refined white sugar (Mitsui Sugar Co., Ltd.
Made, product name: White Zara AA) 543.9 g, Japanese crystal cell
Loin (Asahi Kasei Co., Ltd., trade name: Avicel RC-59
1) Mix 30 g and use as a binder
Ropilcellulose (Shin-Etsu Chemical Co., Ltd., trade name: HPC-
(L) 10g was dissolved in 200ml of distilled water
Fluidized bed granulator (SPRAY-GRA manufactured by Fuji Sangyo Co., Ltd.
Always in NUMATOR MODEL STREA-1)
Granulated by the method. The obtained granules are sized to 30-83
550 g of granules A having a particle size of mesh were obtained. Again this
Those that passed through 83 mesh were passed through the above fluidized bed granulator.
From the same, make granules in the same way, 30-83 mesh classification
Was collected and combined with the granule A obtained above to give a total of 680 g.
Granule B was obtained. Separately, Japanese amino acid acetic acid (Showa Denko
Product name: Glycine "Fine powder") 40g, JP D-Mannito
263g (manufactured by Towa Kasei Co., Ltd., product name: Mannit P)
As a binder,
A solution of 5 g of glucose in 100 ml of distilled water
Granulated in the same manner as Granule A, with a grain size of 30-83 mesh
300 g of granules C were obtained. This granule B343.5g and granule
A mixture containing 156.5 g of CKM and containing RKM hydrate
Lee syrup preparation 500g (1g RKM 100mg strength
Value). This dry syrup formulation is usually 3g
(300 mg titer as RKM) 1 time 3 times a day
Administered to children. Reference Example 4 The following Reference Example 1 was performed using 100 g of RKM hydrate.
680 g of Granule B was obtained in the same manner as in. Separately,
Minoacetic acid 40g, JP D-mannitol 260g
As a binder, Japanese Pharmacopoeia Hydroxypropyl Cellulose 5
g dissolved in 100 ml of distilled water
Cium (product name: Shiroishi Calcium Co., Ltd .: Corocalco W)
B) Granulate in the same manner as Granule A, using a suspension of 3 g
And 300 g of granules C having a particle size of 30 to 83 mesh
Obtained. This granule B343.5g and granule C156.5g
Dry syrup formulation mixed and containing RKM hydrate
500 g (100 mg titer of RKM in 1 g) were obtained.

Claims (5)

[Claims] 1. Formula (1): Rokitamycin monohydrate crystal represented by. 2. The formula (1): wherein rokitamycin is crystallized in an acidic aqueous solution. Method for producing rokitamycin monohydrate crystals represented by 3. The acidic aqueous solution contains p of an organic acid or an inorganic acid.
The method according to claim 2, which is an aqueous solution of H1 to H4. 4. The method according to claim 3, wherein the organic acid is acetic acid or propionic acid. 5. The method according to claim 2, wherein the inorganic acid is phosphoric acid.