The invention deals with novel peptides useful for the therapeutic treatment of infectious diseases caused by pathogenic microorganisms. Included is the peptide FR900156 having the structure: ##STR1## as well as the peptides of the structure ##STR2## wherein R1 is acyl;
Rb1 is hydrogen, methyl, isopropyl, hydroxymethyl, protected hydroxymethyl or benzyl;
R2 is hydrogen, carboxy, protected carboxy, or a group of the formula: ##STR3## wherein Ra2 is mono- or di-carboxy lower alkyl or ar(carboxy) lower alkyl whose aryl moiety may be substituted by hydroxy,
Rb2 is hydrogen or lower alkyl;
Rp and Rq are each hydrogen, carboxy, protected carboxy, with proviso that when one of R2 and Rq is hydrogen, then the other is carboxy or protected carboxy;
Rr is hydrogen or amino protective group; m is an integer 1 to 3, and n is 1 provided that when R1 is benzyloxycarbonyl R51 is methyl, m is an integer 2 and n is an integer 1, then R2 is not protected carboxy, and provided that when R1 is hydrogen, t-butoxycarbonyl or N-acetylmuramyl, Rb1 is methyl, m is an integer 2 and n is an integer 1, then R2 is hydrogen, protected carboxy or a group of the formula: ##STR4## wherein Ra2 is mono- or di-carboxy lower alkyl having 1 and 3 to 6, carbon atoms, α-carboxyethyl, --CH2 --CH2 --COOH, ar(carboxy) lower alkyl whose aryl moiety may be substituted by hydroxy and Rb2 is as defined above, or its pharmaceutically acceptable salt.
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1. A compound of the formula: ##STR379## wherein
R1 is acyl; Rb1 is hydrogen, methyl, isopropyl, hydroxymethyl, protected hydroxymethyl or benzyl; R2 is hydrogen, carboxy, protected carboxy, or a group of the formula: ##STR380## wherein Ra2 is mono-or di-carboxy lower alkyl or ar(carboxy)lower alkyl whose aryl moiety may be substituted by hydroxy, Rb2 is hydrogen or lower alkyl; Rp and Rq are each hydrogen, carboxy, protected carboxy, with proviso that when one of R2 and Rq is hydrogen, then the other is carboxy or protected carboxy; Rr is hydrogen or amino protective group; m is an integer 1 to 3; and n is 1, provided that when R1 is hydrogen, t-butoxycarbonyl or N-acetylmuramyl, Rb1 is methyl, m is an integer 2 and n is an integer 1, then R2 is hydrogen, protected carboxy or a group of the formula: ##STR381## wherein Ra2 is mono- or di-carboxy lower alkyl having 1 and 3 to 6, carbon atoms, α-carboxyethyl, ar(carboxy)lower alkyl whose aryl moiety may be substituted by hydroxy and Rb2 is as defined above, or its pharmaceutically acceptable salt. 2. A compound according to the
R1 is acyl, Rb1 is hydrogen, methyl, isopropyl, hydroxymethyl or benzyl, R2 is hydrogen, carboxy or a group of the formula: ##STR382## wherein Ra2 and Rb2 is as defined in claim 1, Rp and Rq are each hydrogen or carboxy, with proviso that when one of R2 and Rq is hydrogen, then the other is carboxy, and Rr is hydrogen. 3. A compound according to the
n is an integer 1 and m is an integer 1. 4. A compound according to
and m is 1. 5. A compound according to the
n is an integer 1 and m is an integer 2. 6. A compound according to
1 and m is 2. 7. A compound according to the
n is an integer 1 and m is an integer 3. 8. A compound according to
and m is 3. 9. A compound according to the
R2 is hydrogen or protected carboxy, Rp and Rq are each hydrogen or protected carboxy, with proviso that when one of R2 and Rq is hydrogen, then the other is protected carboxy. 10. A compound according to the
carboxy. 11. A compound of the formula: ##STR383## 12. A compound of the formula: ##STR384## 13. A compound of the formula: ##STR385## wherein R1 is an aliphatic acyl radical which is unsubstituted alkanoyl or alkenoyl or is alkanoyl of at least two carbon atoms or alkenoyl substituted by amino, halogen, hydroxy, lower alkoxy, or lower alkanoylamino; Rb1 is hydrogen, methyl, isopropyl, hydroxymethyl, protected hydroxymethyl or benzyl; R2 is hydrogen, carboxy, protected carboxy, or a group of the formula: ##STR386## wherein Ra2 is mono- or di-carboxy lower alkyl or ar(carboxy)lower alkyl whose aryl moiety is unsubstituted or is substituted by hydroxy, Rb2 is hydrogen or lower alkyl; Rp is hydrogen, carboxy, or protected carboxy, Rq is carboxy or protected carboxy; Rr is hydrogen or amino protective group; m is an integer 1 to 3; and n is 1, or its pharmaceutically acceptable salt. 14. A compound of the formula: ##STR387## wherein R1 is acyl, Rb1 is hydrogen, methyl, isopropyl, hydroxymethyl or benzyl, R2 is hydrogen, carboxy or a group of the formula: ##STR388## Ra2 is mono- or di-carboxy lower alkyl or ar(carboxy)lower alkyl whose aryl moiety is unsubstituted or is substituted by hydroxy, Rb2 is hydrogen or lower alkyl; Rp is hydrogen or carboxy; Rq is carboxy; Rr is hydrogen. m is an integer 1 to 3; and n is 1, provided that when R1 is t-butoxycarbonyl or N-acetylmuramyl, Rb1 is methyl, m is an integer 2 and n is an integer 1, then R2 is hydrogen, or a group of the formula: ##STR389## wherein Ra2 is mono- or di-carboxy lower alkyl having 1 and 3 to 6 carbon atoms, -CH2 -CH2 -COOH, ar(-carboxyl)lower alkyl whose aryl moiety is unsubstituted or is substituted by hydroxy and Rb2 is as defined above, or its pharamaceutically acceptable salt. 15. A compound according to the claim 13, wherein n is an integer 1 and m is an integer 1. 16. A compound according to claim 14 wherein n is 1 and m is 1. 17. A compound according to the claim 13, wherein n is an integer 1 and m is an integer 2. 18. A compound according to claim 14, wherein n is 1 and m is 2. 19. A compound according to the claim 13, wherein n is an integer 1 and m is an integer 3. 20. A compound according to claim 14, wherein n is 1 and m is 3. 21. A compound according to the claim 13, wherein R2 is hydrogen or protected carboxy, Rp is hydrogen or protected carboxy, and Rq is protected carboxy. 22. A compound according to the claim 13, wherein the protected carboxy for at least one of R2, Rp and Rq is esterified carboxy. |
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This invention relates to a new peptide. More particularly, this invention relates to a new peptide and the pharmaceutically acceptable salt thereof, which have pharmacological activities, to processes for the preparation thereof and to a new intermediate for preparing the active peptide, and to the pharmaceutical composition comprising the same and a method of use thereof.
I.R. (CHCl3): 3400, 3300, 1730, 1710, 1665 cm-1.
NMR (CDCl3), δ(ppm): 1.36 (3H, d, J=7 Hz), 1.14 (3H, d, J=7 Hz), 1.4-2.0 (4H, m), 2.08 (3H, s), 2.2-2.4 (2H, m), 4.4-4.7 (2H, m), 4.9-5.2 (1H, m), 5.10 (2H, s), 6.96 (1H, d, J=8 Hz), 7.15 (1H, d, J=8 Hz), 7.20 (5H, s), 7.4-8.2 (1H, broad s).
D-Lac(OAc)-L-Ala-γ-D-Glu(α-OBzl)-L)-Boc-(D)-mesoDAP-(D)-NHNH Boc-(L)-GlyOH D-Lac(OAc)-L-Ala-γ-D-Aad(α-OBzl)-(L)-Boc-(D)-mesoDAP-(D) -NHNHBoc-(L)-GlyOH (5) was prepared from D-Lac(OAc)-L-Ala-D-AspOBzl D-Lac(OAc)-L-Ala-D-AadOBzl (3) and Boc-(D)-mesoDAP-(D)-NHNHBoc-(L)-GlyoH (4) in substantially the same manner as that of Example 66.
m.p. 108°-110°C
I.R. (Nujol): 3260, 1725, 1670, 1650, 1630, 1540, 1520 cm-1.
NMR (CDCl3), δ(ppm): 1.40 (3H, d, J=7 Hz), 1.4-2.0 (13H, m), 1.44 (18H, s), 2.10 (3H, s), 2.1-2.5 (2H, m), 3.90 (2H, s), 3.8-4.2 (1H, m), 4.3-4.6 (3H, m), 4.7-5.1 (1H, m), 5.12 (2H, s), 7.30 (5H, s).
D-Lac-L-Ala-γ-D-Glu(α-OH)-(L)-mesoDAP-(L)-GlyOH D-Lac-L-Ala-γ-D-Aad(α-OH)-(L)-mesoDAP-(L)-GlyOH (6) was prepared from D-Lac(OAc)-L-Ala-γ-D-Glu-(α-OBzl)-(L)-Boc-(D)-mesoDAP-(D)-N HNHBoc-(L)-GlyOH D-Lac(OAc)-L-Ala-γ-D-Aad-(α-OBzl)-(L)-Boc-(D)-mesoDAP-(D )-NNHBoc-(L)-GlyOH 1040 (5) in substantially the same manner as that of Step 2 of Example 88.
I.R. (Nujol): 3270, 1720, 1620, 1530 cm-1.
NMR (D2 O), δ(ppm): 1.36 (3H, d, J=7 Hz), 1.43 (3H, d, J=7 Hz), 1.5-2.1 (10H, m), 2.2-2.5 (2H, m), 3.84 (1H, t, J=6 Hz), 3.96 (2H, s), 4.1-4.5 (4H, m).
PAC (1) Step 1 ##STR377##D-Lac(OAc)-L-Ala-γ-Abu-(L)-Boc-(D)-mesoDAP-(D)-NHNHBoc-(L)-GlyOH (3) was prepared in substantially the same manner as that of Example 66.
NMR (CDCl3), δ(ppm): 1.2-2.0 (4H, m), 1.42 (18H, s), 2.0-2.5 (2H, m), 2.15 (3H, s), 3.1-3.4 (2H, m), 3.8-4.1 (2H, m), 4.3-4.7 (1H, m), 4.9-5.2 (1H, m), 5.5-6.0 (2H, m), 7.3-7.6 (2H, m), 7.6-8.1 (2H, m).
D-Lac(OH)-L-Ala-γ-Abu-(L)-mesoDAP-(L)-GlyOH (4) was prepared in substantially the same manner as that of Example 88.
NMR (D2), δ(ppm): 1.39 (3H, d, J=7 Hz), 1.40 (3H, d, J=7 Hz), 1.5-2.1 (8H, m), 2.33 (2H, t, J=7 Hz), 3.21 (2H, t, J=7 Hz), 3.79 (1H, t, J=7 Hz), 3.95 (2H, s), 4.1-4.5 (4H, m).
The following components were prepared in substantially the same manner as Steps 1 and 2 of Example 1, respectively.
PAC (1) Step 1Thienylacetyl-γ-D-Glu(α-OH)-(L)-Boc(D)-mesoDAP-(D)-NHNHBoc.
NMR (DMSO-d6), δ(ppm): 1.36 (18H, s), 1.00-2.33 (10H, m), 3.66 (2H, s), 3.83-4.33 (3H, m), 6.87 (2H, m), 7.27 (1H, m), 7.83-8.66 (3H, m), 9.02 (2H, broad s).
Thienylacetyl-γ-D-Glu(α-OH)-(L)-mesoDAP.
NMR (D2 O), δ(ppm): 1.20-2.60 (10H, m), 3.78 (1H, t, J=7 Hz), 3.86 (2H, s), 4.28 (1H, t, J=7 Hz), 7.04 (2H, m), 7.32 (2H, m).
For fermentation: (1) A vegetative medium 1 (pH 7.0) was prepared from the following ingredients:
| ______________________________________ |
| Vegetative medium 1 |
| ______________________________________ |
| Soluble starch 2% by wt. |
| Gluten meal 1% by wt. |
| Dried yeast 1% by wt. |
| Corn Steep Liquor |
| 1% by wt. |
| Tap water q.s. |
| ______________________________________ |
100 ml. of the medium 1 in each of ten 500 ml. flasks were sterilised in conventional manner and then inoculated with a loopful of culture from a stock slant of Streptomyces olivaceogriceus ATCC 31427. The organism was grown in the medium at 30°C for 48 hours on a shaker.
Into a 30-liter Jar-fermentor, there were placed 20 liters of the vegetative medium 2 prepared from the following ingredients:
| ______________________________________ |
| Vegetative medium 2 |
| ______________________________________ |
| Soluble starch 2% by wt. |
| Cottonseed meal 0.5% by wt. |
| Gluten meal 0.5% by wt. |
| Dried yeast 0.5% by wt. |
| Corn Steep Liquor |
| 0.5% by wt. |
| tap water q.s. |
| ______________________________________ |
The vegetative medium 2 (pH 7.0) was sterilised in a conventional manner and then inoculated a septically with the whole volume of the vegetative inoculum culture prepared above. The organism was grown in the medium 2 at 30°C for 24 hours.
The whole volume of the vegetative inoculum thus prepared was aseptically inoculated into a 2000-liter fermentor, containing 1600 liters of the fermentation medium prepared from the following ingredients:
| ______________________________________ |
| Fermentation medium |
| ______________________________________ |
| Soluble starch 2% by wt. |
| Cottonseed meal 0.5% by wt. |
| Wheat germ 0.5% by wt. |
| Dried yeast 0.25% by wt. |
| Corn Steep Liquor 0.25% by wt. |
| KH2 PO4 0.5% by wt. |
| Na2 HPO4.12 H2 O |
| 0.05% by wt. |
| CoCl2.6 H2 O |
| 1.25 mg/l |
| Tap water q.s. |
| ______________________________________ |
The organism was cultured in the fermentation medium for 72 hours at 30°C During the growth period, the broth was stirred with a propeller operating at 170 r.p.m. and sterile air was passed through the broth at a rate of 1600 liters per minute.
After the fermentation was completed, 20 kg "Radiolite" (trade name, a filter aid material sold by Showa Chemical Company, Japan) was added to the culture broth and the mixture was filtered to remove mycelia. 1600 liters of the filtrate was passed through a column of activated charcoal (800 liters) and then was washed with 1600 liters of water. Elution was carried out with 3000 liters of 50% aqueous acetone and then the eluate was concentrated to a volume of about 600 liters.
The concentrate was passed through a column of DEAE-Sephadex (trade name, made of Pharmacia A.B.) (200 liters) which has previously been buffered with phosphate buffer (pH 6.0). The column was successively washed with 200 liters of water and 200 liters of 0.1 M sodium chloride solution and then eluted with 400 liters of 0.3 M sodium chloride solution. The aqueous eluate was passed through a column of an activated charcoal (200 l), washed with 200 liters of water and then eluted with 400 liters of 50% aqueous acetone. The elutate was concentrated and then freeze-dried to give 800 g of a white powder. The powder was dissolved into 25 liters of water and the solution was passed through a column of CM-Sephadex (H+ form) (25 l). The column was eluted with 25 liters of water and the eluate was concentrated and then freeze-dried to give 33 g. of yellowish white powder. The powder was placed on the top of a column of cellulose (1.2 l). The column was washed with 1000 ml. of 70% aqueous propanol and then eluted with 1000 ml. of 60% aqueous propanol. The eluate was concentrated and freeze-dried to give 4 g. of white powder. The powder was dissolved into 150 ml. of water and then the solution was subjected to column chromatography on Sephadex G-15 (2.8 l).
The column was developed and eluted with water. The active fractions were collected and concentrated and then freeze-dried to give 4 g. of a white powder. The powder dissolved into 25 ml. of water and the solution was subjected to column chromatography on CM-Sephadex (H+ form) (400 ml). The column was developed and eluted with water. The active fractions were collected, concentrated and then freeze-dried to give 40 mg. of the FR-900156 substance in the form white powders (purity:about 70%).
(2) Fermentation was carried out in the same manner as described in Example 94 (1). After the fermentation was completed, 20 kg. "Radiolite" (trade name, a filter aid material sold by Showa Chemical Company) was added to the culture broth and the mixture was filtered to remove mycelia. 1600 liters of the filtrate was passed through a column of activated charcoal (800 liters) and then was washed with 1600 liters of water. Elution was carried out with 3000 liters of 50% aqueous acetone and then the eluate was concentrated to a volume of about 600 liters. The concentrate was passed through a column of DEAE-Sephadex (trade name, made by Pharmacia A.B.) (200 liters) which has previously been buffered with phosphate buffer (pH 6.0). The column was successively washed with 200 liters of water and 200 liters of 0.1 M sodium chloride solution and then eluted with 400 liters of 0.3 M sodium chloride solution. The aqueous eluate was passed through a column of an activated charcoal (200 liters), washed with 200 liters of water and then eluted with 400 liters of 50% aqueous acetone. The eluate was concentrated and then freeze-dried to give 800 g. of white powder. The powder was dissolved into 25 liters of water and the solution was passed through a column of CM-Sephadex (H+ form) (25 liters). The column was eluted with 25 liters of water and the eluate was concentrated and then freeze-dried to give 33 g. of yellowish white powder. The powders were placed on top of a column of cellulose (1.2 liters). The column was washed with 1000 ml. of 70% aqueous propanol and then eluted with 1000 ml. of 60% aqueous propanol. The eluate was concentrated and freeze-dried to give 4 g. of white powder. The powder was dissolved into 300 ml. of water and then passed through a column of DEAE-Sephadex (trade name, made by Pharmacia A.B.) (1.4 liters) which has previously been buffered with phosphate buffer (pH 6.0). the column was washed with 1.5 liters of 0.1 M sodium chloride solution and then eluted with 3 liters of 0.2 M sodium chloride solution. The active fractions were collected and then passed through a column of activated charcoal (300 ml). The column was washed with water and then elutated with 800 ml. of 50% aqueous acetone. The elutate was concentrafted and then freeze-dried to give 700 mg. of white powders. The powders were dissolved into 20 ml. of water and then passed through a column of CM-Sephadex (H+ form) (500 ml). The column was eluted with water and the active fractions were collected, and then concentrated to give 10 ml. of concentrate. The concentrate was subjected to a column chromatography on Sephadex G 15 (500 ml) and developed with water. The active fractions were collected and concentrated and then freeze-dried to give 70 mg. of white powders. The powders were dissolved into 25 ml. of water and subjected to preparative thin layer chromatography on cellulose (made by Eastman Kodak Co.): a developing solvent was mixture of butanol, acetic acid and water (4:1:2). Elution was carried out with 50 ml. of water and the eluate was concentrated and then freeze-dried to give 20 mg of the FR-900156 substance in the form of a white powder.
(3) Fermentation was carried out in the same manner as described in Example 94 (1). After the fermentation was completed, 20 kg. "Radiolite" (trade name, a filter aid material sold by Showa Chemical Company) was added to the culture broth and the mixture was filtered to remove mycelia. 1600 Liters of the filtrate was passed through a column of activated charcoal (800 liters) and then was washed with 1600 liters of water. Elution was carried out with 3000 liters of 50% aqueous acetone and then the eluate was concentrated to a volume of about 600 liters. The concentrate was passed through a column of DEAE-Sephadex (trade name, made by Pharmacia A.B.) (200 liters) which has previously been buffered with phosphate buffer (pH 6.0). The column was successively washed with 200 liters of water and 200 liters of 0.1 M sodium chloride solution and then eluted with 400 liters of 0.3 M sodium chloride solution. The aqueous eluate was passed through a column of an activated charcoal (200 liters), washed with 200 liters of water and then eluted with 400 liters of 50% aqueous acetone. The eluate was concentrated and then freeze-dried to give 800 g. of white powder. The powder was dissolved into 25 liters of water and the solution was passed through a column of CM-Sephadex (H+ form) (25 liters). The column was eluted with 25 liters of water and the eluate was concentrated and then freeze-dried to give 33 g. of yellowish white powder. The powder was placed on top of a column of cellulose (1.2 liters). The column was washed with 1000 ml. of 70% aqueous propanol and then eluted with 1000 ml. of 60% aqueous propanol. The eluate was concentrated and freeze-dried to give 4 g. of white powder. The powder was dissolved into 300 ml. of water and then passed through a column of DEAE-Sephadex (trade name, made by Pharmacia A.B.) (1.4 liters) which has previously been buffered with phosphate buffer (pH 6.0). The column was washed with 1.5 liters of 0.1 M sodium chloride solution and then eluted with 300 liters of 0.2 M sodium chloride solution. The active fractions were collected and then passed through a column of activated charcoal (300 ml). The column was washed with water and then elutated with 800 ml. of 50% aqueous acetone. The eluate was concentrated and then freeze-dried to give 700 mg. of white powders. The powders were dissolved into 20 ml. of water and then passed through a column of CM-Sephadex (H+ form) (500 ml). The column was eluted with water and the active fractions were collected, and then concentrated to give 10 ml. of concentrate. The concentrate was freeze-dried to give 400 mg of a powder. The powder was placed on the top of a column of cellulose (500 ml). Elution was carried out with a mixture of n-butanol, acetic acid and water (4:1:2). The active fractions were collected and freeze-dried to give 200 mg of a powder. The powder was dissolved into 7 ml. of water and then placed on Sephadex G 15 (250 ml).
The active fractions were collected, concentrated and then freeze-dried to give 100 mg of the FR-900156 substance in the form of a white powder.
(4) A white powder of FR-900156 substance obtained by the Example 94 (3) was further purified by conducting repeatedly the above purification means to give a more purified product of FR-900156 substance.
(5) 100 ml of a medium containing corn starch 2% (by wt.), gluten meal 1%, dried yeast 1% and corn steep liquor 1% were poured into each of ten 500 ml flasks and sterlised in a conventional manner and then inoculated with a loopful of culture from a stock slant of Streptomyces violaceus ATCC 31481.
The organism was grown in the medium at 30°C for 48 hours on a shaker.
Into a 30-liter Jar-fermentor, there were placed 20-liters of the same medium as above. The medium was sterilised in a conventional manner and then inoculated aseptically with the whole volume of the inoculum culture prepared above. The organism was grown in the medium at 30°C for 30 hours.
The whole volume of the inoculum thus prepared was aseptically inoculated into a 400-liters fermentor, containing 320 liters of the medium (pH 6.5) containing soluble starch 2% (by wt.), gluten meal 1%, cottonseed meal 1% and sodium sulfate (10 hydrates) 2%.
The organism was cultured in the medium at 30°C for 72 hours. During the growth period, the broth was stirred with a propeller operating at 170 r.p.m. and sterile air was passed through the broth at a rate of 320 liters per minute. After the fermentation was complete, 4 kg of "Radiolite" was added to the cultured broth and the mixture was filtered to remove mycelia. 300 liters of the filtrate was passed through a column of activated charcoal (150 liters) and then washed with 300 liters of water. Elution was carried out with 600 liters of 50% aqueous acetone and then the eluate was concentrated to a volume of about 120 liters.
The concentrate was passed through a column of DEAE-Sephadex (30 liters) which has previously been buffered with phosphate buffer (pH 6.0). The column was successively washed with 30 liters of water and 30 liters of 0.1 M sodium chloride and then eluted with 0.3 M sodium chloride. The eluate (60 liters) was passed through a column of an activated charcoal (30 liters), washed with 60 liters of water and then eluted with 60 liters of 50% aqueous acetone. The eluate was freeze-dried to give 120 g of a white powder. The powder was dissolved in 4 liters of water and the solution was passed through a column of CM-Sephadex (H+ form) (14 liters). The column was eluted with water and the eluate was concentrated and then freeze-dried to give 20 g of a powder. The powder was placed on the top of the column of cellulose. Elution was carried out with an aqueous propanol and the active fractions were collected and freeze-dried to give 5 g of a powder. The powder was dissolved in 500 ml of water and the solution was passed through a column of DEAE-Sephadex (1.3 liters) which has previously been buffered with phosphate buffer (pH 6). The column was washed with 0.1 M sodium chloride and eluted with 0.2 M sodium chloride. The active fractions were collected and passed through a column of an activated charcoal (900 ml). The column was washed with water and eluted with 50% aqueous acetone (200 ml). The eluate was concentrated and freeze-dried to give 1 g of a powder.
The powder was dissolved in 300 ml of water and the solution was passed through a column of CM-Sephadex (H+ form) (250 ml). Thc column was developed and eluted with water. The active fractions were collected, concentrated and then freeze-dried to give 800 mg of a powder. The powder was dissolved in 20 ml of water and the solution was mixed with a small amount of cellulose. The mixture was subjected to column chromatograph on cellulose. The column was washed successively with 100 ml of acetone and 300 ml of mixture of n-butanol:acetic acid:water (4:1:1) and then developed and eluted with a mixture of n-butanol:acetic acid:water (4:1:2) (1 liter). The active fractions were collected and freeze-dried to give 30 mg of a powder. The powder was dissolved in 20 ml of wter and the solution was passed through a column of Sephadex G 15 (250 ml). The column was developed and eluted with water and the active fractions were collected and then freeze-dried to give 5 mg of FR-900156 substance.
For the pharmaceutical composition:
(1) Preparation for injection
The required quantities of the FR-900156 substance were distributed into vials, each containing 500 mg of the active ingredient. The vials were sealed hermetically to exclude bacteria. Whenever the vial is required for use, 2 ml of sterile distilled water for injection is added to the vial and then the aqueous solution is administered by injection.
(2) Preparation of tablet
A suitable formulation for a tablet consists of the following mixture.
| ______________________________________ |
| FR-900156 substance |
| 200 mg |
| Mannitol 400 mg |
| Starch 50 mg |
| Magnesium stearate |
| 10 mg |
| ______________________________________ |
(3) Preparation of capsule
| ______________________________________ |
| FR-900156 substance |
| 300 mg |
| Magnesium stearate |
| 15 mg |
| ______________________________________ |
The above ingredients were mixed and then inserted into a hard gelatin capsule in a conventional manner.
Hashimoto, Masashi, Nakaguchi, Osamu, Kohsaka, Masanobu, Imanaka, Hiroshi, Okada, Satoshi, Tanaka, Hirokazu, Kitaura, Yoshihiko, Hemmi, Keiji, Takeno, Hidekazu, Aratani, Matsuhiko, Iguchi, Eiko, Aoki, Hatsuo, Kuroda, Yoshio
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| Executed on | Assignor | Assignee | Conveyance | Frame | Reel | Doc |
| May 18 1984 | Fujisawa Pharaceutical Co., Ltd. | (assignment on the face of the patent) | / |
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