7-aminocephem derivative compounds
10308666 ยท 2019-06-04
Assignee
Inventors
Cpc classification
International classification
Abstract
This disclosure provides methods of making certain 7-aminocephem derivatives useful in the manufacture of cephalosporin antibiotic compounds.
Claims
1. A process for making a compound of the formula (II): ##STR00005## comprising the steps of: (a) forming a reaction mixture comprising phosphorous pentachloride (PCl.sub.5) and pyridine; (b) adding a compound of the formula (IIa) to the reaction mixture: ##STR00006## (c) adding isobutyl alcohol; (d) adding a solvent comprising water; (e) adding ethyl acetate; and (f) obtaining the compound of the formula (II).
2. The process of claim 1, wherein step (a) comprises: (1) combining PCl.sub.5 and dichloromethane; and (2) adding pyridine.
3. The process of claim 2, wherein step (1) comprises: (i) combining 2.0 equivalents PCl.sub.5 and 6.0 volumes dichloromethane while maintaining the temperature below 30 C.; and (ii) adjusting the temperature to 10 to 30 C. and agitating.
4. The process of claim 2, wherein step (2) comprises: (i) adjusting the temperature to 10 to 5 C.; (ii) adding 2.0 equivalents of pyridine while maintaining the temperature at 10 to 5 C.; and (iii) adjusting the temperature to 5 to 0 C. and agitating.
5. The process of claim 1, wherein step (b) comprises: (1) adding 1.0 equivalent compound (IIa) to the reaction mixture of step (a) while maintaining the temperature of the mixture at 10 to 0 C.; and (2) stirring the mixture at 10 to 0 C.
6. The process of claim 5, wherein the mixture of step (2) comprises 5.0% compound (IIa) relative to compound (II)+compound (IIa).
7. The process of claim 1, wherein step (c) comprises: (1) adjusting the temperature of the mixture to 20 to 10 C.; (2) adding 2.0 volumes of isobutyl alcohol dropwise while maintaining the temperature of the mixture at 20 to 5 C.; and (3) agitating the mixture at 10 to 0 C.
8. The process of claim 1, wherein step (d) comprises: (1) adding a solvent comprising water to the mixture of step (c) while maintaining the temperature of the mixture at 10 to 5 C.; (2) agitating the mixture at 5 to 5 C.; and (3) separating a lower organic phase from the mixture and collecting the lower organic phase.
9. The process of claim 8, wherein the solvent comprising water is a mixture of ethanol and water.
10. The process of claim 8, which further comprises: (4) adding dichloromethane to the mixture at 10 to 0 C.; (5) agitating the mixture at 10 to 0 C.; and (6) separating a lower organic phase from the mixture and collecting the lower organic phase.
11. The process of claim 1, wherein step (e) comprises: (1) concentrating the organic phase to 4 to 5 volumes; (2) adjusting the temperature to 10 to 5 C.; (3) adding 2.1 volumes of ethyl acetate dropwise while maintaining the temperature of the mixture at 10 to 5 C.; and (4) agitating the mixture at 10 to 5 C.
12. The process of claim 11, wherein step (e) further comprises: (5) observing the formation of a precipitate; (6) concentrating the organic phase to 4 to 5 volumes; (7) adjusting the temperature of the mixture to 10 to 5 C.; (8) adding 2.1 volumes of ethyl acetate while maintaining the temperature of the mixture at 10 to 8 C.; and (9) agitating the mixture at 10 to 5 C.
13. The process of claim 1, wherein step (f) comprises: (1) centrifuging the mixture to produce a cake; and (2) collecting the cake.
14. The process of claim 13, wherein step (f) further comprises: (3) adding 2.2 volumes of ethyl acetate; (4) adjusting the temperature to 10 to 5 C.; (5) agitating at 10 to 5 C.; (6) centrifuging to produce a cake; (7) collecting the cake; and (8) drying the cake at 5 to 20 C.
Description
BRIEF DESCRIPTION OF THE DRAWINGS
(1)
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DETAILED DESCRIPTION
(14) Compound (II) is also known as ACLE.HCl and ((6R,7R)-3-(chloromethyl)-2-(((4-methoxybenzyl) oxy)carbonyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-en-7-aminium chloride) and has the structure shown below.
(15) Provided herein is a method of making compound (II):
(16) ##STR00002##
(17) In one aspect, the method comprises the steps of: (a) forming a reaction mixture comprising phosphorous pentachloride (PCl.sub.5) and pyridine; (b) adding compound (IIa) to the reaction mixture; (c) adding isobutyl alcohol; (d) adding a solvent comprising water; (e) adding ethyl acetate; and (f) obtaining compound (II). The method can be performed in one or more steps. Unless otherwise indicated, one or more steps may be combined in alternative embodiments of the methods disclosed herein.
(18) In one embodiment, step (a) comprises: (1) combining PCl.sub.5 and dichloromethane; and (2) adding pyridine. Step (1) can comprise the steps of: (i) combining 2.0 equivalents PCl.sub.5 and 6.0 volumes dichloromethane while maintaining the temperature below 30 C.; and (ii) adjusting the temperature to 10 to 30 C. and agitating. Step (2) can comprise (i) adjusting the temperature to 10 to 5 C.; (ii) adding 2.0 equivalents of pyridine while maintaining the temperature at 10 to 5 C.; and (iii) adjusting the temperature to 5 to 0 C. and agitating.
(19) In one embodiment, step (b) comprises: (1) adding 1.0 equivalent compound (IIa) to the reaction mixture of step (a) while maintaining the temperature of the mixture at 10 to 0 C.; and (2) stirring the mixture at 10 to 0 C. At the completion of step (b), the mixture of step (2) can comprise 5.0% compound (IIa) relative to compound (II)+compound (IIa).
(20) In one embodiment, step (c) comprises: (1) adjusting the temperature of the mixture to 20 to 10 C.; (2) adding 2.0 volumes of isobutyl alcohol drop-wise while maintaining the temperature of the mixture at 20 to 5 C.; and (3) agitating the mixture at 10 to 0 C.
(21) In one embodiment, step (d) comprises: (1) adding a solvent comprising water to the mixture of step (c) while maintaining the temperature of the mixture at 10 to 5 C.; (2) agitating the mixture at 5 to 5 C.; and (3) separating a lower organic phase from the mixture and collecting the lower organic phase. In one embodiment of step (d), the solvent comprising water is a mixture of ethanol and water. In another embodiment, step (d) further comprises: (4) adding dichloromethane to the mixture at 10 to 0 C.; (5) agitating the mixture at 10 to 0 C.; and (6) separating a lower organic phase from the mixture and collecting the lower organic phase.
(22) In one embodiment, step (e) comprises: (1) concentrating the organic phase to 4 to 5 volumes; (2) adjusting the temperature to 10 to 5 C.; (3) adding 2.1 volumes of ethyl acetate drop-wise while maintaining the temperature of the mixture at 10 to 5 C.; and (4) agitating the mixture at 10 to 5 C. In another embodiment, step (e) further comprises: (5) observing the formation of a precipitate; (6) concentrating the organic phase to 4 to 5 volumes; (7) adjusting the temperature of the mixture to 10 to 5 C.; (8) adding 2.1 volumes of ethyl acetate while maintaining the temperature of the mixture at 10 to 8 C.; and (9) agitating the mixture at 10 to 5 C.
(23) In one embodiment, step (f) comprises: (1) centrifuging the mixture to produce a cake; and (2) collecting the cake. In another embodiment, step (f) further comprises: (3) adding 2.2 volumes of ethyl acetate; (4) adjusting the temperature to 10 to 5 C.; (5) agitating at 10 to 5 C.; (6) centrifuging to produce a cake; (7) collecting the cake; and (8) drying the cake at 5 to 20 C.
(24) Referring to
(25) ##STR00003##
(1) Preparation of Compound (II)
(26) DCM (2135.4 kg, 6.0 vol.) and PCl.sub.5 (230.1 kg, 2.0 eq) were charged to reactor 1, while maintaining the batch temperature below 30 C. The temperature of the batch in reactor 1 was adjusted to 10 to 30 C. and the batch was agitated at 10 to 30 C. for 0.5 to 1.0 hour. The batch temperature was adjusted to 10 to 5 C. Then pyridine (89.4 kg, 2.0 eq) was charged to the batch in reactor 1 drop-wise at 10 to 5 C. The batch temperature was adjusted to 5 to 0 C. and the batch was agitated for 30-40 minutes.
(27) Compound (IIa) (269.8 kg, 1.0 eq) was charged to reactor 1 in portions at 10 to 0 C. The batch was stirred at 10 to 0 C. for 1 to 2 hrs, or until 5.0% compound (IIa) was detected by HPLC. Charging compound (IIa) at 10 to 0 C. is critical to prevent epimerization of the CN(H) bond. See Studies on the Epimerization of compound (II), below).
(28) The batch temperature was adjusted to 20 to 10 C. The adjustment of temperature to 20 to 10 is critical in order to prevent reformation of compound (IIa) in the following step.
(29) The batch was then charged with isobutyl alcohol (446.0 kg, 2.0 vol) drop-wise, while maintaining the batch temperature at 20 to 5 C. The use of isobutyl alcohol prevents the formation of impurities such as ethyl chloride. The batch was agitated at 10 to 0 C. for 0.5 to 2.0 hours. HPLC: (IIa)/[(II)+(IIa)]5.0% (Method AM-C11030404-A-01).
(30) (2) Work-Up of Compound (II)
(31) Water (612 kg, 2.3 vol) and EtOH (206.4 kg, 1.0 vol) was charged into reactor 3 and was agitated for at least 5 min. The temperature of reactor 3 was adjusted to 5 to 5 C. The batch in reactor 1 was then transferred to reactor 2. A portion of the solution of water and ethanol (H.sub.2O/EtOH: 3:1) (551.4 kg, 2.0 vol) prepared was charged into reactor 2, while maintaining the batch at 10 to 5 C. The contents of reactor 2 were stirred at 5 to 5 C. for at least 10 min. The agitation was stopped and the phases were allowed to separate for at least 10 minutes. The lower organic phase is transferred to reactor 1. DCM (148.0 kg, 0.41 vol) was charged to reactor 2 and the batch was agitated for 10 to 20 min at 10 to 0 C. The agitation was stopped and the phases were allowed to separate at 10 to 0 C. for at least 10 minutes and then the lower organic layer was transferred to reactor 1. The rest of the solution of water and ethanol (H.sub.2O/EtOH: 3:1) (270 kg, 1.0 vol) was charged into the batch in reactor 1 at 10 to 0 C. The contents of reactor 1 was agitated at 10 to 0 C. for 20 to 30 minutes. The agitation was then stopped and the phases were allowed to separate for at least 10 minutes. HPLC: (IIa)/[(II)+(IIa)]5.0% (Method AM-C11030404-A-01). The contents of reactor 2 were agitated at 10 to 0 C. for 5 to 10 min. The agitation was then stopped and the phases were allowed to separate at 10 to 0 C. for at least 10 minutes. The lower organic phase was transferred to reactor 1. The solvents, reagents, stoichiometry, temperatures and reaction times of step (2) were selected to increase yield and reaction efficiency.
(32) (3) Isolation of Compound (II)
(33) The organic phase in reactor 1 was concentrated at 25 C. jacket temperature to 1076 to 1345 L (4 to 5 vol) under reduced pressure. Then the batch temperature was adjusted to 10 to 5 C. Ethyl Acetate (513.0 kg, 2.1 vol) was charged drop-wise to the batch at 10 to 5 C. for 2 to 4 hours. Then the batch was agitated at 10 to 5 C. for 5 to 6 hours. A precipitate forms. The agitation was continued until 2.0% of compound (II) was detected in the supernatant by HPLC. If compound 2 is >2% then the batch temperature was adjusted to 5 to 5 C. and concentrated at 25 C. jacket temperature to 1076 to 1345 L (4 to 5 vol) under reduced pressure. Then the temperature was adjusted to 10 to 5 C. Ethyl Acetate (513.0 kg, 2.1 vol) was charged drop-wise to the batch at 10 to 8 C. over the course of 2 to 4 hours. Then the batch was agitated at 10 to 5 C. for 5 to 10 hours. HPLC: Compound (II) in supernatant 2.0% (Method AM-C11030404-A-01).
(34) The batch was centrifuged and the wet cake was collected. Ethyl Acetate (540.0 kg, 2.2 vol) was charged into reactor 1. The wet cake was transferred into reactor 1. The temperature in reactor 1 was adjusted to 10 to 5 C. and the batch was agitated at 10 to 5 C. for 2 to 5 hours. The batch was centrifuged and the wet cake was collected. HPLC: Pyridine 0.1%; (IIa) 0.1%; total purity 97% (Method AM-C11030404-A-01).
(35) The solid was dried under reduced pressure at 5 to 20 C. for 40 to 70 hours. The batch was deemed dry when: water 0.4%; EtOAc 5000 ppm; Ethanol 5000 ppm; DCM 6000 ppm; i-BuOH 5000 ppm. Yield 198.7 kg (87% molar yield) of compound (II) with 99% purity using HPLC method AM-C11030404-A-01.
(36) TABLE-US-00001 TABLE 1 Materials for the preparation of Compound (II) MW. Weight Mass Material (g/mol) (kg) Moles Equivalents (w/w) Density (IIa) 486.97 269.8 0.55 1.0 1.00 PCl.sub.5 208.24 230.1 1.11 2.0 0.85 Pyridine 79.10 89.4 1.13 2.0 0.33 DCM 2135.4 7.92 1.33 i-Butyl 446.0 1.65 0.81 alcohol H.sub.2O 612 2.27 1.00 EtOH 206.4 0.77 0.80 DCM 148.0 0.55 1.33 EtOAc 513.0 2.11 0.90 EtOAc 540.0 2.00 0.90 (II) 405.30 224.5 (theoretical) (II) 405.30 198.7 (isolated)
Studies on the Epimerization of Compound (II)
(37) The purpose of this study was to test the robustness of the manufacturing process for making compound (II) (ACLE-HCl) from compound (IIa) (GCLE) and to better understand the origin of impurity (peak 8) in the Ceftolozane drug substance. This impurity was observed at 0.03% in the Ceftolozane drug substance. The impurity was isolated by Prep-HPLC and its structure was determined to be the 7-epimer of Ceftolozane by HRMS and NMR analysis.
(38) The starting material ACLE-HCl was prepared from GCLE by treatment with phosphorus pentachloride and pyridine in dichloromethane, followed by alcoholysis with isobutanol. As shown in
(39) I. Preparation of ACLE-HCl at Elevated Temperature
(40) TABLE-US-00002 TABLE A List of Materials Ratio Material MW Weight (g) mMoles eq. (w/w) Density GCLE 486.97 10.0 20.54 1 1X PCl.sub.5 208.24 8.55 41.07 2.0 0.86X pyridine 79.1 3.30 45.18 2.0 0.33X DCM 80.0 8X 1.33 iso-butyl alcohol 16.2 1.62X 0.81 H.sub.2O 10.0 1X 1.0 EtOH 36.0 3.06X 0.8 EtOAc 108.0 10.8X 0.9 ACLE-HCl 405.3 8.32 (theoretical) ACLE-HCl 405.3 5.83 (isolated) Isolated yield 70%
II. Process Steps
1. Charged 80.0 g (6 V) DCM into Reactor 1 (R1).
2. Charged 8.55 g (2 eq) PCl.sub.5 into R1 at room temperature (RT).
3. Stirred R1 at 20 to 25 C. for 0.5 h.
4. Cooled R1 to 10 to 0 C.
5. Added 3.3 g (2.0 eq) pyridine into R1 drop-wise at 10 to 5 C.
6. Stirred R1 for 0.5 h at 5 to 5 C.
7. Charged 10.0 g GCLE into R1 at 10 to 0 C.
8. Warmed to RT.
9. Stirred at 35 to 40 C. for 1 h using warm water.
10. Cooled R1 to 20 to 10 C.
11. Added 16.2 g isobutyl alcohol into R1 drop-wise at 20 to 10 C.
12. Stirred R1 at 10 to 0 C. for 2 h.
13. Charged 20.0 g (2.0) EtOH:H.sub.2O (1:3) into R1 at 10 to 5 C.
14. Stirred R1 at 10 to 5 C. for 1 h.
15. Allowed phase separation at 10 to 0 C.
16. Aqueous layer was extracted with 5 g DCM, stirred for 30 min at 10 to 0 C.
17. Allowed phase separation at 10 to 0 C.
18. Combined organic layer in R2 charged with 10 g EtOH:H.sub.2O (1:3) at 10 to 0 C.
19. Stirred R2 for 30 min at 10 to 0 C.
20. Allowed phase separation at 10 to 0 C.
21. To the organic layer was charged 10 g water.
22. Stirred for 30 min at 10 to 0 C.
23. Allowed phase separation at 10 to 0 C.
24. Organic layer was filtered and washed with DCM (2 mL).
25. Organic layer was concentrated to 28 g using evaporator at 20 C. (25 C.).
26. Charged EtOAc (18 g) at 10 to 5 C.
27. The slurry was stirred for 5 h at 10 to 5 C.
28. Organic layer was concentrated to 28 g using evaporator at 20 C. (25 C.).
29. The slurry was stirred for 5 h at 10 to 5 C.
30. The slurry was cooled to 10 to 0 C., and stirred for 1 h.
31. Filtration to collect the solid, washed with EtOAc (3 mL).
32. Dried solid under vacuum with N.sub.2 purge overnight (5.83 g, yield 70%, purity 9495%).
III. Preparation of (Epi-II) (7-Epi-ACLE-HCl)
(41) With reference to
(42) Preparation of p-nitrobenzaldehyde imine (1): To a stirred suspension of ACLE-HCl (22.3 g, 55.0 mmol) in ethyl acetate (200 mL) and water (75 mL) was added 1 N aq. NaOH (100 mL) at 0 C. After addition, the mixture was stirred for 30 min. to give a clear two-phase solution. The EtOAc layer was separated, washed with water (275 mL) and dried with Na.sub.2SO.sub.4 (50 g). To above suspension was added p-nitrobenzaldehyde (9.8 g, 65.0 mmol). The mixture was stirred at R. T. for 2 h, then diluted with DCM (400 mL). The Na.sub.2SO.sub.4 was removed by filtration and washed with DCM. The filtrate was concentrated to give a paste, which was filtered and washed with 10% EtOAc in hexanes to give a yellowish solid. The solid was dried under in-house vacuum with N.sub.2 purge to provide the imine (23.6 g, 85.5%). 1H NMR (CDCl.sub.3) 8.73 (1H, d, J=1.8 Hz), 8.30 (2H, d, J=8.7 Hz), 7.97 (2H, d, J=8.7 Hz), 7.39 (2H, d, J=8.7 Hz), 6.93 (2H, d, J=8.7 Hz), 5.50 (1H, dd, J=1.8, 5.1 Hz), 5.27 (2H, s), 5.22 (1H, d, J=5.1 Hz), 4.59 (1H, d, J=12.0 Hz), 4.42 (1H, d, J=12.0 Hz), 3.84 (3H, s), 3.71 (1H, d, J=18.3 Hz), 3.48 (1H, d, J=18.3 Hz).
Epimerization: To a solution of imine 1 (10.34 g, 20.6 mmol) in THF (200 mL) was added NEt.sub.3 (100 mg, 1 mmol) at 0 C. Within 2 min., the reaction was quenched by adding acetic acid (200 mg), diluted with EtOAc (50 mL) and water (50 mL). The EtOAc layer was separated, washed with water and brine, dried over Na.sub.2SO.sub.4. After filtration, the filtrate was concentrated to give a mixture of the imines (13 g, 7:71:2.5). The two isomeric imines can be separated by flash chromatography, even though they will decompose during the chromotagraphy. For 7 imine: 1H NMR (CDCl.sub.3) 8.54 (1H, d, J=1.2 Hz), 8.29 (2H, d, J=8.7 Hz), 7.93 (2H, d, J=8.7 Hz), 7.39 (2H, d, J=8.7 Hz), 6.90 (2H, d, J=8.7 Hz), 5.34 (1H, d, J=12.0 Hz), 5.24 (1H, d, J=12.0 Hz), 4.96 (1H, d, J=2.1 Hz), 4.88 (1H, dd, J=1.2, 2.1 Hz), 4.48 (1H, d, J=12.0 Hz), 4.39 (1H, d, J=12.0 Hz), 3.80 (3H, s), 3.73 (1H, d, J=18.0 Hz), 3.50 (1H, d, J=18.0 Hz).
Preparation of 7-epi-ACLE-HCl: To a mixture of 2, 4-dinitrophenylhydrazine (3.96 g, 20 mmol), p-toluenesulfonic acid monohydrate (3.80 g, 20 mmol) in ethanol (800 mL) was added the above prepared imine mixture (13 g) in CHCl.sub.3 (60 mL) at RT. After addition, the mixture was stirred at RT for 4 hrs until the completion of the reaction indicated by TLC. The orange precipitate was removed by filtration, washed with ethanol (20 mL). The filtrate was put into the fridge overnight. The precipitate (2.1 g) was collected by filtration, washed with ethanol (20 mL). 1H NMR showed that the precipitate was ACLE-p-TSA salt. The filtrate was then concentrated to about 400 mL and put into the fridge overnight. The precipitate (0.6 g) was collected by filtration, washed with ethanol (10 mL). The mother liquor was concentrated to give a residue (7.0 g), which was mixed with EtOAc (200 mL) and water (30 mL). To above mixture, 1 N aq. NaOH (30 mL) was added at 0 C. and the mixture was stirred for 30 min at 0 C. to become a clear two-phase solution. The EtOAc layer was separated, washed with water, brine and dried over Na.sub.2SO.sub.4. After filtration, the filtrate was concentrated to about 100 mL, then 2 N HCl in ether (20 mL) was added at 0 C. and the mixture was stirred at 0 C. for half an hour. The solid was collected by filtration, washed with EtOAc and dried in vacuum to give a brown solid (2.7 g, 7-epi-ACLE-HCl:ACLE-HCl2.5:1). The above solid (100 mg) was crystallized with methanol/acetonitrile to give (20 mg) pretty pure 7-epi-ACLE-HCl. 1H NMR (DMSO-d6) 8.98 (3H, b), 7.39 (2H, d, J=8.0 Hz), 6.95 (2H, d, J=8.0 Hz), 5.30 (1H, d, J=11.6 Hz), 5.21 (1H, d, J=11.6 Hz), 5.04 (1H, s), 4.77 (1H, s), 4.48 (1H, d, J=11.6 Hz), 4.42 (1H, d, J=11.6 Hz), 3.85 (1H, d, J=18.4 Hz), 3.77 (3H, s), 3.56 (1H, d, J=18.4 Hz) (
For ACLE-p-TSA salt: 1H NMR (DMSO-d6) 8.84 (3H, b), 7.48 (2H, d, J=7.6 Hz), 7.37 (2H, d, J=7.6 Hz), 7.11 (2H, d, J=7.6 Hz), 6.94 (2H, d, J=7.6 Hz), 5.185.28 (4H, m), 4.59 (1H, d, J=11.6 Hz), 4.49 (1H, d, J=11.6 Hz), 3.82 (1H, d, J=17.6 Hz), 3.76 (3H, s), 3.69 (1H, d, J=17.6 Hz), 2.29 (3H, s).
IV. HPLC conditions
HPLC column: YMC Pack-ODS-AQ, 3 m, 150 mm4.6 mm
Injection Volume: 10+l
Column Temp: 202 C.
Autosampler Temp: 4+ C.
Wavelength: 254 nm
Mobile Phase A: Buffer Ammonium Acetate Solution: Dissolve 4.0 g of NH4OAc in 0.9 L of water in a 1 L volumetric flask, using a calibrated pH meter, adjust the pH to 3.50.05 with acetic acid and dilute to volume with water. Mix well by magnetic stirrer or by inversion. Mix 450 mL of buffer ammonium acetate solution and 550 mL CAN. Mix well to become the mobile phase A solution.
Mobile Phase B: Acetonitrile
Flow rate: 1.0 mL/min
Run Time: 30 min
Mode: Gradient (see Table B)
(43) TABLE-US-00003 TABLE B Time (min) % A % B 0.00 100 0 12.00 100 0 22.00 40 60 23.00 100 0 30.00 100 0
V. Results and Discussion
(44) The ACLE-HCl prepared at elevated temperature, as described above, was analyzed by HPLC and LCMS. As seen in the HPLC trace, the product showed a similar profile as the in-house standard except for a new peak at 4.29 (
(45) Control of the reaction temperature after the charge of GCLE is important in the manufacturing of ACLE-HCl from GCLE. When the reaction temperature was raised from 100 C. to 3540 C., a small amount of 7-epi-ACLE-HCl (about 23%) was observed in the product. 7-epi-ACLE-HCl was not detected in the in-house standard of ACLE-HCl (Table C).
(46) TABLE-US-00004 TABLE C Effect of Temperature on the Ration of 7-epi-ACLE-HCl to ACLE-HCl Temperature 7-epi-ACLE-HCl (% AUC) ACLE-HCl (% AUC) 10 to 0 C. 0 100 35 to 40 C. 2.8 97.2
Characterization of Compound (II)
(1) HPLC Method AM-C11030404-A-01: Assay by external standard analysis with a gradient, reversed-phase (RP) HPLC method using a YMC-Pack-ODS-AQ (1504.6 mm i.d., 3 m) or equivalent column. Mobile phase A is a mixture of ammonium acetate in water (4 g/L) adjusted to pH 3.5 and acetonitrile (v/v 45:55), and mobile phase B is acetonitrile. UV detection is at 254 nm.
(2) Optical rotation: Analysis is performed on a 10 mg/mL solution in dimethylformamide at 589 nm at 200.5 C. Acceptance criterion for optical rotation of ACLE.HCl is 58 to 47 which is supported by the batch analysis mean of 9 batches plus and minus three standard deviations. Optical rotation values from the 9 batches ranged from 54.2 to 50.7.
(3) Water content of ACLE.HCl was measured by Karl Fischer titration.
(4) FTIR: The IR absorption spectrum (KBr dispersion) was recorded in the range of 4000 to 650 cm 1.
(5) Impurities: Dichloro-PMB ester (IIb) and 7-Epi-ACLE.HCl (epi-II).
(47) The most abundant impurity in ACLE.HCl produced by the above procedure is the compound Dichloro-PMB ester (see
(48) 7-Epi-ACLE.HCl (epi-II) is described above.
(49) ##STR00004##
(6) X-Ray Powder Diffraction.
(50) Provided herein is a crystal form of compound (II). In one embodiment, the crystal form of compound (II) exhibits an X-ray powder diffraction spectrum having characteristic peaks at substantially the same angles as the pattern of
(51) Synthetic Compositions
(52) Compound (II) is a useful intermediates in the production of antibiotics, particularly ceftolozane, and salts thereof. Compositions comprising compound (II) are provided herein. Also provided are compositions produced or occurring during the methods of making compound (II).
(53) The following composition may be produced during method 2: a composition comprising compounds (IIa) and (II); a composition comprising compounds (II) and (IIb); and a composition comprising compounds (II) and (epi-II).
(54) TABLE-US-00005 TABLE 2 Analytical Test Results Analytical Target Typical Analytical Test Method Results Result Consumption of (IIa) HPLC 5.0% 4.9% Consumption of (IIa) HPLC 5.0% 1.8% Consumption of (IIa) HPLC Report 2.0% Residue of (II) in supernatant HPLC 2.0% 0.4% Purity of (II) in wet cake HPLC 97% 99% Residue of (IIa) in wet cake HPLC 0.1% 0.07% Pyridine in wet cake HPLC 0.1% <0.05% KF in (II) during drying KF 0.4% 0.1% EtOAc in (II) during drying GC 5000 ppm 39959 ppm Ethanol in (II) during drying GC 5000 ppm 620 ppm DCM in (II) during drying GC 6000 ppm 3780 ppm i-BuOH in (II) during drying GC 5000 ppm 567 ppm
(55) TABLE-US-00006 TABLE 3 Batch Analysis Data for ACLEHCl: Optical Rotation Batch # Optical Rotation () 1 51.0 2 50.7 3 51.7 4 52.7 5 52.9 6 51.5 7 53.3 8 54.2 9 54.0