TW201028427A - New crystal polymorphs of ceftobiprole - Google Patents

Abstract

A compound of formula (I) in a crystalline form, which has a X-ray diffraction pattern comprising, when measured using Cu-K α 1 radiation, a diffraction line, having a relative intensity (rI) of 100 percent at about 11.6, 12.9, 14.7, 22.0, 23.1 or 24.5, given in degrees 2 Theta [2 θ ]. Five corresponding polymorphic crystal forms (Polymorph A; B; C; D or E) have been characterized by means of their XRPD and have been isolated.

Description

201028427 VI. INSTRUCTIONS OF THE INVENTION: TECHNICAL FIELD OF THE INVENTION The present invention relates to a compound of formula (I):

0 (I) It is also known as "Ceftobiprole" (CAS No. 09467-5 2-7)' and is described in, for example, ΕΡ-Α-0 849 269 & US-A-5,981 , broad-spectrum antibiotics in 5 19 . The derivative of the scorpion scorpion having an improved solubility in an aqueous medium is Ceftobiprole Medocaril according to the following formula.

It has been described, for example, in W〇-A 99/65920. A method for preparing ceftizodol medecali is described, for example, in w〇_A 〇1/9〇111, and includes ceftizox and 5-methyl-2-oxo-[L3] carbonate Reaction of oxol-4-ylmethyl group 4-decyl-phenyl vinegar. Therefore, the compound of formula (1) (head-holding) is important not only as an antibiotic itself, but also as a potential intermediate for the manufacture of derivative antibiotics such as ceftizodol medecali. 145413.doc 201028427 [Prior Art] A method of preparing a compound of formula (I) is described in Ep_A_〇 849 269. In this preparation method, (6R,7R)_7_[(Z)_2(5-amino-[12,4]thiazol-3-yl)-2-diphenylfluorenyloxyimine·B is prepared first.醯Amino]_8_oxo-3_[(E)-(R)-2-oxo-[1,3']bipyrrolidine_3_ylidenemethyl]_5thiazepine-bicycle [4_2.0] Oct-2-ene-2-carboxylic acid, and subsequently reconverted to a compound of formula (1), as specifically described in Example n" of ΕΡ-Α-0 849 269. However, the products obtained by this method have not been used for industrial practical operations. In particular, it has been found that the stability of the compound of formula (I) is too low for continuous use because the formula (I) initially present in the sample prepared according to ΕΡ-Α-0 849 209 The compound has almost broken down in half within 4 weeks. In addition to this inconsistent shelf life, it has been found that the purity of the material produced according to ΕΡ-Α-0 849 269 (i.e., the initial percentage in the compound of formula (1)) is only about 85 〇/〇 at maximum, which is not satisfaction. Therefore, there is a need to substantially improve the initial purity and long-term stability of the material obtained according to ΕΡ-Α-0 849 269, so that the compound of the formula (1) can be applied to all the usual medical operations. Conventional methods for proper improvement include aggressive purification of the starting cephalosporin material and the use of a suitable stabilizer compound to obtain a sufficient shelf life of the starting material. Beta-endamine antibiotics are known to be a compound that is sensitive to hydrolytical degradation and are not susceptible to day-to-day. (Y. Ikeda et al., Chem. Pharm. Bull. 56(10) 1406-1411 (2008)). Furthermore, it is known that only specific polymorphs of the 0-endoamine antibiotic show sufficient stability due to the binding of water molecules in the crystal lattice. (Y. Takeuchi et al., Chem. Pharm. Bull 41 (11) 1998- 145413.doc -5· 201028427 2002 (1993)) ° [Summary of the Invention] However, surprisingly, a specific polycrystal of the compound of the formula (1) has been found. The type (hereinafter referred to as polymorph D) exhibits such greatly improved purity and stability against decomposition so that the above problems can be easily solved. Furthermore, it has been found herein that the compounds of formula (I) exist in a variety of polymorphic forms which have now been isolated and characterized by X-ray powder diffraction (XRPD). Daytime is defined as the ability of a pharmaceutical material to have two or more crystalline phases' which have different molecular arrangements and/or conformations in the crystal lattice. In another aspect, the amorphous solid consists of a disordered molecular arrangement and does not have a distinguishable lattice. Polymorphs can have different chemical and physical properties such as melting point, chemical reactivity, 'apparent solubility, dissolution rate, optical and electrical properties, vapor pressure and density. These characteristics may directly affect the processing of pharmaceutical raw materials and the quality/performance of pharmaceutical products, such as bioavailability, and especially stability. In particular, metastable pharmaceutical solids often change crystal structure over time in response to changes in conditions (such as temperature or humidity, and/or process, etc.). The standard for defining the presence of polymorphs is that they are generally proved to be different crystal structures by their X-ray diffraction pattern. Microscopic 'thermal analysis methods and solid-state NMR are commonly used as supporting sources for information obtained by supporting 乂-ray diffraction analysis. Preparation of the newly discovered polymorphs A, b, c, sputum and cephalosporin in the following description, which facilitates the preparation of a medicament comprising cefotaxime itself, and the preparation of a cephalosporin-containing derivative (eg, Department of cephalosporin tromethide ^ 145413.doc 201028427 Lee) of medicine. In particular, 'the newly discovered cefotaxol polymorph D as defined herein is unexpected compared to the cefotaxol amorphous form known in the prior art (as obtained from ΕΡ-Α-0 849 269) Significantly improve the stability and initial purity of the cefdolo substance. When measured using Cu-Κα! radiation, the new polymorph has an X-ray diffraction pattern containing high-intensity ray (peak), specifically, at about 116 or 12.9 or 14.7 or 22.0 or 23.1 or 24· Between 5 and 2 degrees [2 Θ] has a relative intensity (rl) of ray around 1〇〇%. The estimated variability of the 2 Θ measurement is $+/_0·2 degrees. More specifically, the present invention relates to a cephalosporol of one of the polycrystalline crystalline forms (A, Β, C, D or Ε type) indicated below, which is measured using Cu-Koh radiation. 'X-ray diffraction pattern with characteristic diffraction rays (expressed in 2 Θ [2Θ]) containing each polymorph as shown below: Type A B type C type D type E type 9.7 7.9 5.2 8.4 4.6 13.4 10.3 9.0 9.9 8.0 15.1 10.5 11.6 12.0 9.3 15.6 11.9 12.8 14.5 11.5 18.8 12.9 13.8 15.4 12.3 19.4 14.4 15.6 16.9 14.7 23.1 15.3 17.3 18.2 18.3 24.0 16.4 20.3 18.7 18.7 24.3 17.0 23.0 19.8 20.5 25.1 18.8 23.9 21.6 21.9 26.8 20.0 22.7 22.9 27.2 21.8 23.9 — 23.3 22.0 24.5 24.7 24.1 25.2 26.0 25.3 26.7 145413.doc 201028427 The ray is shown to have a relative intensity of η Wang less than 20% based on the strongest peak of each diffraction pattern. Those skilled in the art will appreciate that the relative intensity of the diffraction peaks of a particular compound may vary considerably depending on, for example, the particle size of the sample and the technical and/or device-related parameters used to record the diffraction pattern. . (See, for example, the illustrations shown in Figures 2 and 7, which can be understood when aligning the peak intensity at 2 degrees 84 degrees. The diffraction angles of the peaks generally vary little and generally exhibit a variability of up to 0.2 degrees 2 。. The new polymorphs A, Β, C, 〇 and Ε, when used in the measurement of radiation, include, in particular, the following X-ray diffraction pattern with a diffraction peak of relative intensity (rI) of 4% or higher:

Type A 2 Θ rl 9.7 m 13.4 m 15.1 m 15.6 m 18.8 s 23.1 vs 24.0 m 24.3 m 26.8 m Type B 2 Θ rl m _Ι〇3__ mmm vs msmm 18.8 mmm 22.0 vs 24.1 m 253 273 mm D type 2Θ rl 8.4 S 9.9 m 16.9 m 24.0 mc type 2Θ rl 5.2 m 9.0 m 11.6 vs 12.8 m 13.8 m 15.6 m — 17.3 m 20.3 m 23.0 m 23.9 m

E type 2Θ rl 4.6 S 8.0 m 11.5 m 12.3 m 145413.doc * 8 - 201028427 In the above table, the relative intensities (rl) of the ray are classified as follows: Symbol rl [%] vs 100 s <100 to 85

m < 85 to 40 w < 40 The X-ray diffraction pattern of the crystalline form mentioned may also show other peaks, especially peaks having a relative intensity of less than 40%. In any recorded X-ray image, depending on the exact recording conditions, one or more weaker ray rays may not be found, but they cannot be completely excluded, as one or more of these low-intensity reflections may be Reflection caused by impurities contained in the measured sample. A particularly preferred embodiment of the invention is:

24.5 VS 26.7 m 14.7 VS 18.3 m 18.7 m 20.5 m 22.9 m 23.3 m 24.7 m 26.0 s • Polycrystalline crystalline form D (polymorph D) with head-holding Tolo, • Polycrystalline crystalline form A (polymorph A) And its polycrystalline crystalline form E (polymorph E). The best line D type. The most stable cephalosporin polymorph found in polymorph D is and appears to be thermodynamically stable under normal conditions. The purity of the cefotaxod polymorph D is much higher than that of the prior art, and its tendency to decompose during storage (also under high temperature and increased relative humidity) is significantly reduced, as in the present application. Example 8 shows. The polymorphs A, B, C, D and E according to the present invention are readily distinguished by their differential X-ray powder diffraction pattern. [Embodiment] The crystalline ceftotropine crystal form of the present invention can be prepared as follows: Polymorph A can be obtained, for example, by (6匕71^_7_[(2;)_2_(5-amino-[1] , 2,4]thiadiazol-3-yl)-2-trisuccinyloxyiminoamino-acetamido]-8_oxo-3-[(E)-(R)-2-oxo -[1,3,]bipyrrolidine_3_ylidenemethyl]_5thia-1-aza-bicyclo[4.2.0]oct-2-ene-2-hypoacid dihydrochloride N Ο

The isolated solids were collected from the solids obtained by chromatography and obtained from the pure compound of formula (1), as described in more detail in Example 3 or in a similar manner. Polymorph D can be obtained as follows: (for example) at a temperature below about the generation by Taopu 7 training_2_(5_Sixian, 4]κ3 yl)_2 trityloxyiminoaminoacetamido] _8_Gas soil...-Subunit methyl]-5-Sulfur Si _Corrinic acid and trifluoroacetic acid and triethylbicyclo [4.2.〇] octene I olefin I guess the mixture, then add three gas 疋 B reaction 'Add water, ethyl acetate, B

Mem hose column, water is used to transfer the separated aqueous phase to elute the product, vacuum to remove "= about 9:1 acetic acid: 混合物.4 mixture guess" by adjusting the pH to about 3, so that the production 145413 .doc 201028427 Precipitates and separates the polymorph Form D of the compound of formula (I) obtained. It is described in more detail in Example 6. Polymorphs B, C and E may be as in Examples 4, 5 or in the present application, respectively. Description of 7 or obtained in a similar manner. Example 1 (6R,7R)-7-[(Z)-2-(5-Amino-[1,2,4]thiadiazol-3-yl)-2- Trityloxyimido-acetamido]oxo-3-[(E)-(R)-2-oxo-[1,3,]bipyrrolidin-3-ylidenemethyl] -5-thia-1-aza-bicyclo[4.2.0]oct-2-en-2-carboxyindole hydrazine dihydrochloride is prepared according to, for example, ΕΡ-Α-0 849 269 (6R, 7R)-3-[(E)-(R)-1·-allyloxycarbonyl-2-oxo-[1,3,]bipyrrolidin-3-ylidenemethyl]-7-[( Z)-2-(5-Amino-[1,2,4]thiadiazol-3-yl)_2-triphenylhydrazinyloxyimino _ acetamido]-8-oxo-5- Hexa-1-aza-bicyclo[4.2.0]oct-2-ene-2-teric acid (4.12 g, 4.79 mmol) was dissolved in 280 ml of dichloromethane suspension to add 1.87 ml ( 7. 66 mmol) N,0-bis(trimethyldecyl)acetamide, which in turn forms a dialling solution' followed by 84 mg (0.12 mmol) of bis-(triphenylphosphine) _ di-gasification (II) , 5"48 ml (95.8 mmol) acetic acid and 11.7 ml (44.1

Methyl) tributyltin hydrogenated and stirred at room temperature for 40 minutes. After adding a few drops of water, the suspension was poured into 丨500 ml of diethyl ether (containing 2 ml of 6 M hydrochloric acid in diethyl ether). The suspension was stirred for 2 hours and the product was collected by filtration. Yield: 4.04 g (99%) of beige solid IR (KBr) 1781, 1659 cm*1 MS (ISP) 777.4 (M+H)+. 1454I3.doc -11 - 201028427 Example 2 (611,711)-7-[(2)-2-(5-Amino-[1,2,4]thiadiazol-3-yl)-2-triphenyl Methyl oxyimido-acetamido]-8-oxo-3-[(E)-(R)-2-oxo-[1,3,]bi-pyrrolidine-3·subunit A 5-5-thia-1-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylic acid under nitrogen, 13.2 g of palladium(II) acetate (0.059 mol), 46.2 g Phenylphosphine (0.17 mol) was dissolved in 4_2 L of tetrahydrofuran, and 116 ml of citric acid (3 mol) was added at room temperature. The resulting solution is cooled to i to i 0〇c, and (6R,7R)-3-[(E)-(R)- is prepared, for example, according to EP-AO 849 269, within 2 Q to 10 minutes. l·-Allyloxycarbonyl_2_oxoindole, "11-pyrrol-3-phenylidenemethyl]-7-[(Z)-2-(5-amino-[H4]thiadiazole _3_基)_2•trityloxyimino-acetamido]_8_oxo_5_thiazepine-aza-bicyclo[4.2.0]oct-2-ene-2- A solution of carboxylic acid (704 g, 〇·73 mol) in όL·tetrahydrofuran. Then add 4 〇〇ml of tributyltinhydride (1.51 mol) in 2 to 10 minutes at -5 to -HTC. And the mixture was stirred for 5 to 30 minutes. The resulting precipitate was collected by filtration, washed with 3 5 L of tetrahydrofuran, and dried overnight at room temperature under a pressure of less than 1 〇bar. Yield: 450 g Example 3 (cefofole _ Polymorphic form a): 17.89 g of (6R,7R)_7_[(Z)_2_(5_Amino-[124]. Dioxin% soil) 2 _ 甲基 methyl oxyimino group _ acetamino group Oxygen _3_[(e)_(r)_2· oxo-[ι,3·]linked subunit fyl]_5_thia"aza-bicyclo[4.2.0] bis_2_thrace _2_Resinic acid dihydrochloride is added to 2〇〇 (4) formic acid and stirred 145413.do c •12·

201028427 Mix for 1 hour. The suspension was added to 2 L of ethyl acetate under stirring and stirred for additional 30 min. The solid was isolated by filtration, washed with ethyl acetate and diethyl ether and dried in vacuo. The beige solid was purified by MCI gel (Mitsubishi, CHP-20P) chromatography using a gradient of water and increasing acetonitrile concentration. All fractions containing the pure product were combined and evaporated to remove acetonitrile. The precipitate was collected by filtration and dried under vacuum. Yield: 7.56 g of pale yellow crystals IR (MIR) 1764, 1658 cm1 MS (ISP) 535.1 (M+H)+. 1H-NMR (250 MHz): δ [ppm] in DMSO-d6: 4H, 3.60, d, J=17 Hz 3.79, d, J=17 Hz; 6H, 5.07, d, J=8 Hz; 7H, 5.75 , dd, J-5 Hz, J=8 Hz; NH, 9.46, d, J= 8 Hz; NH2, 8.05, s; OH, 12.0, s br; 3Ή, 7.30, s; 4Ή, 5Ή 4"H 5HH 1.92-2.16, 2.5-3.5, m; 1"H, 10.3 s br; 3"H, 4.66, m = XRP D ray (recorded with Cu-Και light) is shown in the following table i. Table 1: Type A °2Θ rl Γ%1 9.7 52 13.4 56 15.1 47 15.6 41 ~ 17.7 22 18.8 92 19.4 36 23.1 100 24.0 62 Α type, continued table °2Θ 24.3 47 25.1 31 26.8 41 27.2 30 31.6 21 34.0 23 36.0 20 Example 4 (cefotabole-polymorph B): 145413.doc -13· 201028427 1 g of (6R,7R)-7-[(Z)-2-(5-amino-[ι,2 , 4] 嗟dis-3-yl)_2-trityloxyimino-acetamido]_8-oxo_3_[(£)_(尺)_2_氧代_ [1,3 ']bipyrrolidin-3-ylidenemethyl]-5-thia-^aza-bicyclo[4_2.〇]octyl-2-ene-2-carboxylic acid dihydrochloride in 90% of 10 ml Stir in formic acid for 1 hour, then pour into 100 ml of ethyl acetate' and stir for an additional hour. The solid was collected by filtration and added to 8 ml of distilled water to give a homogeneous suspension. Add acetonitrile (a few drops) until a coloring solution is obtained. This solution was purified on 1 〇〇 ml of MCI gel (Mitsubishi, CHP-20P) eluting with water (10 min) and acetonitrile gradient (1 min) with 90% water / 10% acetonitrile. The fractions containing the pure product were concentrated in vacuo and the resulting solid was collected by filtration and dried under high vacuum. Yield: 270 mg of beige solid IR (MIR), MS (ISP) and 1H-NMR (250 MHz). The X-ray XRPD ray (using Cu-Koii, although recorded) is shown in Table 2 below. Table 2 Type B °2Θ rl Γ%1 7.9 44 10.3 59 10.5 61 11.9 69 12.9 100 14.4 85 15.3 87 16.4 76 17.0 75 18.8 74 20.0 69 Β type, continued table °2Θ rl Γ%1 21.8 70 22.0 100 24.1 82 25.3 56 26.7 38 27.3 41 30.2 32 31.6 27 32.7 26 34.7 31 Example 5 (cefotabole-polymorph Q: 145413.doc • 14· 201028427 at 0.6°C to 0.6 ml of triethyldecane and 10 ml of trifluoro 1.62 g of (6R,7R)-7-[(Z)-2-(5-amino-indole, 2,4]oxadiazol-3-yl)-2-tricarboxylate was added to the acetic acid mixture. Benzoyloxyimino-acetamido]-8•oxo(R)-2-oxo-[1,3,]bipyrrolidin-3-ylidenemethyl]-5-thiazepine; [_Aza-bicyclo[4.2.0]oct-2-ene-2-carboxylic acid. The mixture was continuously stirred at 0 ° C for 3 Torr. Under a vacuum of suction, the trifluoroacetic acid was evaporated and was high. The residue was dried under vacuum. The residue was dissolved in water and ethyl acetate, and the aqueous phase was concentrated. The mixture was adjusted to pH 3. The suspension was stirred at 0 ° C for 3 hours and the product was collected by hydrazine. The solid was dissolved in 15 ml IN HC1 and 1.5 ml of acetonitrile. The solution was passed through a 5 g bed of MCI gel (Mitsubishi, CHP-20P) and the gel was washed with a 12:1 mixture of IN HC1 and acetonitrile 9:1. The filtrate was concentrated. The acetonitrile was removed and concentrated NaOH was added to adjust the pH to 3.0. The resulting suspension was condensed to a volume of about 40 ml and stirred for 2 hours at 〇 ° C. The solid was collected by filtration and dried to constant weight. Yield: 0.76 g. Solid IR (MIR), MS (ISP) and 1H-NMR (250 MHz) are shown in Example 3. XRPD diffraction (recorded with Cu-Κα丨 radiation) is shown in Table 3 below. Table 3 Type C °2Θ rl [%] 5.2 85 9.0 85 11.6 100 12.8 72 Type C, continued table °2Θ rl Γ%1 17.3 67 20.3 69 23.0 66 23.9 65 145413.doc 201028427 13.8 65 — 15.6 75 Example 6 (cefotabole-polymorph D): 1.6 g of (6R,7R)_7_[(Z)_2_(5-amino-[12 4]thiadiazol-3-yl)-2-trityloxyimido group at -10 °C -acetamido]_8_oxo_3_ [(E)-(R)-2-oxo-[1,3']bipyrrolidine_3_ylidenemethyl]_5_thiazepine Aza-a-cyclo[4.2.0]oct-2-ene-2-acidic acid was added to 5 mi of trifluoroacetic acid. Immediately at 0 C, diethyl decane (〇·3 mi) was added and the mixture was stirred for 3 〇 minutes. Approximately 4 g of ice, 25 ml of ethyl acetate and 25 ml of water were added: a mixture of 9:1:0.4 of acetonitrile:trifluoroacetic acid. Separate the clear phases. The aqueous phase was transferred to a column containing 17 g of Mci-gel (Mitsubishi, CHP-20P) adjusted with a 9:1 mixture of water and acetonitrile. The product was eluted from the bed with water: a 9:1:0.4 mixture of acetonitrile:trifluoroacetic acid. The product was contained in 4 about 12 ml each of the fractions, and the acetonitrile was removed under suction vacuum and concentrated to adjust to pH 3.00. The resulting precipitate was stirred at rt for 4 hours and collected by filtration. Yield: 0.65 g of off-white crystalline solid IR (MIR), MS (ISP) and 1H-NMR (250 MHz). The XRPD ray (recorded by shot) is shown in Table 4 below. Table 4 D type °2Θ rl Γ%1 8.4 41.5 9.9 44.5 12.0 20.9 14.5 21.5 14.9 15.3 15.4 31.1 Type D, continued table °2Θ nr%l 18.7 22.4 19.8 25.4 20.7 15 21.6 25 22.7 26.1 24.0 56.1 145413.doc -16- 201028427 16.9 51.1 18.2 21.6 24.6 _ 100 26.7 37.1 Example 7 (head holding - polymorph E): at -20C, within 10 to 20 minutes, to 82 〇 ml of trifluoroacetic acid and 72 ml of triethyl The decane is dissolved in a mixture of 2_24 L of di-methane and 3 80 g of (6R,7R)-7-[(Z)-2-(5-amino-[丨- thiazolidine-3- Benzyl)-2-benzhydryloxyimino-acetamido]_8-oxo_3_[(e)-(R)-2-oxo-[1,3']bipyrrole-3 - subunit fluorenyl]-5-thiato_ι_aza-bicyclo[4.indol]oct-2-ene-2-carboxylic acid. The resulting suspension was stirred for a further 15 minutes at -15 °C. The lower layer was separated from the biphasic solution, and the upper layer (dichloromethane) was extracted with a mixture of 290 ml of trifluoroacetic acid and 290 ml of water. The lower layer was combined with the extract and eluted with 290 ml of acetonitrile in 2_9 L of distilled water. At 13〇 g Dicalite

The combined solution was filtered on a Speedex and washed twice with a solution of trifluoroacetic acid: B. Water 1.2:3:2 4 solution. The combined filtrate was chromatographed on a 4 kg MCI-gel (Mitsubishi, CHP-20P) eluting with a solution of 20 l of trifluoroacetic acid: acetonitrile: water 1.2:3:24. The main dissolved fraction of the product was evaporated to a volume of 6 to 8 L at 40 to 45 ° C, 100 to 40 mbar. The resulting pale yellow solution was diluted with 1.9 L of distilled water. The pH was adjusted to 3.6 to 3.8 by cooling to 0 ° C using 1.1 L of 28% sodium hydroxide solution and >pH 3 of dilute sodium hydroxide solution (0.2 M). From 0 to 5. (: After stirring for another 3 to 4 hours, the suspension was filtered, washed with 1.9 L of cold distilled water (2 ° C) and 1.9 L of cold acetone (2 ° C) at 40 ° C, < 1 bar The obtained crystals were dried for 14.5 hours. Yield: 172 g of crystalline IR (MIR), MS (ISP) and 1H-NMR (250 MHz). See Example 3. 145413.doc -17· 201028427 XRPD ray (with (:11-1) ^〇11radiation record) is shown in Table 5. Table 5 E type °2Θ rl [%] 4.6 90 8.0 45 9.3 39 11.5 79 12.3 77 14.7 100 18.3 44 18.7 58 20.5 80 21.9 39 Type E, continued table °2Θ rl Γ°/〇] 21.9 37 22.9 56 23.3 43 24.7 42 26.0 89 27.3 36 29.2 37 32.1 32 32.9 33 Example 8 After storage for 0, 1, 2 and 4 weeks at 40 ° C / 75 ° / 〇rh, compare The stability of the cephalosporin batch of batch D (batch 07R0004) obtained according to Example 6 and the amorphous cephalosporin batch (batch 966/15) obtained according to EP-A 0 849 269. Monitoring by HPLC Purity/content and crystallographicity by XRPD. HPLC measurement: Instrument: HPLC system: quaternary pump module (Waters Alliance 2695) DAD detector (Waters Alliance 2996) Sampler (Waters Alliance 2695) column oven (Waters Alliance 2695) authorization control and integration software

Mettler Toledo MT5. Can be 0.5 ° /. Tolerance and reading error of 1/5 / 145413.doc -18- 201028427 Tolerance ratio Weigh the balance of the required compound. Detailed HPLC method: Flow rate: 1.2 mL/min Mobile phase A: 3/97/0.1, ACN/water/TFA (v/v/v) Mobile phase B: 1000/1, ACN/TFA (v/v) tube Column: Prontosil 120-3-C1 8 (75x4_6 mm), 3 μιη particle size

Column temperature: 30 ° C

Autosampler: 5°C ❹ Injection: 10 kL Detection: UV or DAD λ=229 nm Gradient program for this method:

Time (minutes) % A %B 0.0 1.0 10.0 25.0 42.0 46.0 48.0 50.0 50.4 55.0 ο ο ο ο 0 0 8 2 0 0 ο ο IX 11 00 00 -ό sw IX ΙΑ ο ο ο 8 0 0 5 5 14 7 9 9 ο ο Materials · Acetonitrile: Mercklichrosolv gradient grade. Water: Merck lichrosolv gradient grade. Sample solvent. Water: Add 400 mL of M.P.A. to a 500 mL graduated cylinder and replenish the volume with acetonitrile. Add 15 mL of trifluoroacetic acid. 145413.doc •19- 201028427 Sample. Lot number/sample weight (mg) Dilution volume (mL) 07R0004/t=0 HPLC 99.17 100 07R0004/t=l HPLC 69.105 100 07R0004/t=2 HPLC 101.13 100 07R0004/t=4 HPLC 95.745 100 966/15/t=0 HPLC 106.125 100 966/15/t=l HPLC 102.41 100 966/15/t=2 HPLC 99.605 100 966/15/t=4 HPLC 99.905 100 Preparation of mobile phase:

Mobile phase A Add 30 mL of acetonitrile to a 1 L graduated cylinder. Replenish the volume with water. Add 1 mL of trifluoroacetic acid.

Mobile phase B Add 1 mL of acetonitrile to 1 mL of trifluoroacetic acid. Calculate the content of the ceftotropine sample, that is, analyze the content of cefotaxod in the sample: *100 content (sample) area sample.a, w standard goods a β -*-· +3⁄4 standard area #准* AWg Sample Results: This crystalline form (polymorph D of cefotaxime) is significantly more stable than its amorphous form (see Table, Table 7 and especially Table 8). After storage, the purity of the crystalline ceftotropine (polymorph D) remained unchanged, while the purity of the corresponding amorphous compound decreased by 25%. As used herein, "purity" refers to the area % of the cefotaxime peak in the HPLC chart. After storage for 4 weeks, the content of the crystalline cefotaxod (polymorph D) (as defined above) decreased by only 3%, while the content of the cefotaxol amorphous sample decreased by nearly 145413.doc -20- 201028427 40%. Table 6: Cefotaxol batch number 07R0004 (crystallization) stability batch number 07R0004 07R0004 07R0004 07R0004 t=0 t-1 t=2 t=4 Area area % Area area % Area area % Area area % Cephaloprol 18863604 94.85 12467521 95.32 18128158 95.09 17553775 95.26 Total amount 19881227 100 13079165 100 19065150 100 18427124 100 Content 90-1 85.4 84.9 86.8 Table 7: Cefotaxol batch 966/15 (amorphous) stability batch number 966/15 966/15 966/15 966/15 t =0 t=lt=2 t=4 Area Area % Area Area % Area Area % Area Area % Cephaloprol 18620782 86.18 13795137 76.06 11906214 73.35 8171189 59.03 Total 20447099 100 18136925 100 16231855 100 13843238 100 Contains Dong 78.6 64.0 59.6 38.7

Table 8: Summary Relative Percent Change Batch Number Crystal Form t=0 t=lt=2 t=4 Purity change in HPLC (area%) w 07R0004 Polymorph D 0.0 0.5 0.2 0.4 966/15 Amorphous 0.0 10.1 -12.8 -27.2 Content change (cefotabole%)|b| 07R0004 Polymorph D 0.0 -4.6 -5.2 -3.3 966/15 Amorphous 0.0 -14.6 -19.0 -39.9 [a] in purity (time point) - purity ( t=0) Calculate [b] Calculate the graphical representation of the data in Table 5 and Figure 6 in Table 8 by content (time point)_content (t=〇). As seen in these XRPDs (see Table 9 below), the cefotaxol polymorph did not change the crystalline form during the test. The XRPD pattern of the cefotaxazole lot 07R0004 corresponds to the crystalline form D. Table 9: Comparison of storage t = 0 weeks and t = 4 weeks, cephalosporin batch number 07R0004 2 peak intensity-21 - 145413.doc 201028427 t=0 t=0 t==4 -----1 t= 4 (.) Strength Relative Strength Strength Relative Strength CPS _ [%1 CPS [%] 8.4 48.8 41.5 42.6 34.9 9.9 52.2 44.5 51.4 42.1 —12.0 24.6 20.9 19.5 16.0 __J4.5 25.2 21.5 46.5 38.1_ _____14.9 18.0 15.3 33.2 27.2 36.5 31.1 61.5 50.5 a __ —_16.9 60.0 51.1 47.4 38.8 a 18.2 25.4 21.6 25.3 20.7_ 26.3 22.4 30.6 25.1 ___19.8 29.9 25.4 27.7 22.7 a 20.7 17.6 —~1Ϊ0 24.9 20.4 ___2L6 --- 29.4 25.0 30.3 24.9 22.7 30.6 26.1 26.8 21.9 -^2^0 65.9 56.1 93.0 76.3_ 24.6 117.5 100.0 122.0 100.0_一_ ___26.7 43.5 37.1 62.4 51.1 NMR spectrum of cefotaxazole _ lot 966/15 after storage for 4 weeks due to content degradation The impurity signal is displayed. BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 shows the XPrd spectrum of polymorphic form A of cefotaxod (recorded by Cu-Koh radiation); Figure 2 shows the xPrd spectrum of polymorphic 头 of ceftizox (recorded by Cu-Kq radiation) Figure 3 shows the XPRD spectrum of polymorph E of cefotaxod (recorded with Cu_Kai radiation); Figure 4 shows the XPRD spectroscopy of amorphous cephalosporin obtained according to EP_A 〇 849 269 (using Cu-Ka丨 light shot) Record); Figure 5 shows the difference between the purity of the head of the polymorphous type D and the corresponding amorphous material after 4 weeks; Figure 6 shows the polymorphic type of the wafer in the head holding tray. ^The corresponding amorphous substance of U-Qi Uxing has changed in percentage of 4 weeks; 145413.doc -22- 201028427

Figure 7 shows the XPRD spectrum of the newly prepared cefotaxod polymorph D batch No. 07/R0004 (Reference Example 8 (t = 0 weeks); and Figure 8 shows the cefotaxol polymorph D batch No. 07/R0004 (Reference example) 8. XPRD spectrum after 4 weeks of storage (t=4 weeks) at a temperature of 40 ° C and a relative humidity (rh of 75%). 145413.doc •23·

Claims (1)

201028427 VII. Scope of application for patents: 1. A compound of formula (I) in crystalline form (Ceftobiprole): It has a diffraction ray having a relative intensity (rl) of 1% at 2 Θ degrees [2Θ] of about u 6 or 12 9 or 14.7 or 22.0 or 23·1 or 24.5 when measured using Cu-Κα! radiation. X-ray diffraction pattern. 2. A compound of the formula (I) according to claim 1 which is a polycrystalline crystalline form of the type A, B, C, D or E hereinafter, which is measured using Cu-Kal radiation. There is an X-ray diffraction pattern comprising diffraction peaks of respective polymorphs represented by 2 degrees [2 Θ] as follows, and the diffraction peaks indicated have 2 相对 with respect to the strongest peak of each diffraction pattern. Relative strength of % and higher: Type A - 2Θ 9.7 13.4 15.1 15.6 17.7 18.8 19.4 23.1 24.0 24.3 25.1 26.8 27.2 31.6 ~3ί〇Β Type C 2Θ 2Θ 7.9 5.2 10.3 9.0 10.5 11.6 11.9 12.8 12.9 13.8 14.4 15.6 15.3 17.3 16.4 20.3 17.0 23.0 18.8 23.9 20.0 21.8 22.0 24.1 25.3 __ D type ~ type ~ 2Θ 2Θ 8.4 4.6 — 9.9 8.0 12.0 9.3 14.5 11.5 14,9 12.3 15.4 14.7 16.9 18.3 18.2 18.7 18.7 20.5 19.8 21.9 20.7 22.9 _ 21.6 23.3 ~~ 22.7 24.7 ~~ 24.0 26.0 24.6 ~~273 ~~ 145413.doc 201028427 36.0 26.7 26.7 29.2 27.3 32.1 30.2 32.9 31.6 32.7 34.7 3. As in the case of claim 2 (I a compound having an X-ray diffraction pattern as measured using Cu-KtM radiation, comprising diffraction peaks of polymorphs as indicated below and expressed in degrees 20 [2q], the diffractions indicated The peak has a relative intensity of 40% and higher with respect to the strongest peak of each diffraction pattern: Type A B type ~ cm 2 Θ rl 2 Θ rl 2 Θ rT 9.7 m 7.9 m ~ 52~ Γ 1 — s_____ 13.4 m 10.3 m 15.1 m 10.5 m ~ Iu~~~ m — 15.6 m 11.9 m ~ΪΙΓ~~ Vi 18.8 s 12.9 vs 13.8 ill 23.1 vs 14.4 m ~Κβ~~ m 24.0 m 15.3 s "ΊτγΗ —_5L__ 24.3 m 16.4 m ~2〇3~~ 26.8 m 17.0 m ~2l〇~~~ 18.8 m 23.9 m 20.0 m I· 21.8 m 22.0 vs 24.1 m 25.3 m 27.3 m D type E type 2Θ rl 2Θ Π 8.4 s 4.6 1 s 9.9 m 8.0 ---1 m 16.9 m 11.5 m 24.0 m 12.3 m 24.5 vs 14.7 ^vs" 26.7 m 18.3 m 18.7 m 20.5 m 22.9 m 23.3 m 24.7 m 26.0 s where the relative intensities (rl) of the ray rays can be classified as follows: 145413.doc D type, ^ °2Θ rl Γ%1 18.7 -r~s 1 22.4 19.8 25.4 20.7 15 21.6 25 22.7 26.1 24.0 56.1 24.6 100 26.7 37.1 201028427 Symbol rl [%] VS 100 S <100 to 85 m <85 to 40 W < 40 4. A type A, D or E as in any of claims 2 to 3. The compound of the formula (I) is a polycrystalline knot. The compound of the formula (1) of any one of claims 2 to 4 is a type D and E. The compound of the formula (1) according to any one of claims 2 to 5, which is in the form of polycrystalline crystal form D. 7. The compound of the formula (1) according to any one of claims 2 to 6 which has a diffraction pattern of X-ray powder comprising the following ray (recorded with ~Και light): °2Θ 8.4 41.5 9.9 44.5 12.0 20.9 14.5 21.5 15.3 15.4 31.1 16.9 51.1 _ 18.2 21.6 8. A compound of the formula (1) according to any one of the claims, which is a polycrystalline crucible type A. Next day 9. A compound according to any one of claims 1 to 4, which has a diffraction pattern of X-ray powder comprising the following ray (recorded with a centimeter-radiation): B type °2Θ Π Γ%1 7.9 44 10.3 59 10.5 61 11.9 69 12.9 100 Β type, performance table °2Θ η γ%ι 22.0 100 24.1 82 25.3 56 26.7 38 27.3 "41 145413.doc 201028427 ίο 11. If request E is 14.4 ~~153~~ -~ _85 __87 16.4 76 17.0 18.8 20.0 21.8 -__70 The compound of the term 'is a multi-E. 1 is a polycrystalline crystalline form of a compound according to any one of claims 1 to 4 or 1 , an X-ray powder diffraction pattern of a rewinding ray (using Cu-Ka, having a lower 歹|J ___, radiation) Record): °2Θ rl [%1 4.6 8.0 45 9.3 11.5 79 12.3 77 14.7 100 18.3 44 18.7 — 58 20.5 80 -_2L9__ 39 ~ 12. A compound for the preparation of the compound of the formula (1) (cefofol:, '° crystal polymorph) N, ΟΗ (I) by: (6R,7R)_7 like) o-amino-[124] thiazol-3-yl)_2_trityloxyimine at a temperature below about 〇 °C Ethylaminoamine]_8oxo_3_[(Ε)-(Κ)_2-oxo-oxime, 3·]bipyrrolidin-3-ylidenemethyl]_5_thiazepine 145413.doc - 4- 201028427 Hetero-bicyclo[4.2.0]oct-2-ene-2-carboxylic acid is reacted with trifluoroacetic acid and triethyldecane; a mixture of water, ethyl acetate, acetonitrile and additional trifluoroacetic acid is added; The aqueous phase is separated from the obtained mixture and the separated aqueous phase is transferred to a MCI gel column; the product is eluted with a mixture of about 9:1:0_4 of acetonitrile:trifluoroacetic acid; acetonitrile is removed under vacuum; The cefotaxol polymorph product is precipitated by adjusting the pH to about 3; and is isolated from the polymorph. 13. A method of preparing a crystalline polymorph of a compound of formula (1) (cefotabole): Its system: (I) The amine group of the formula: ^, 2, 4] 嚷 嗤 3 3 3 3 3 3 3 笨 ] ] ] ] [ [ [ [ [ [ [ [ [ [ [ [ [ [ [ [ [ [ [ [ [ [ [ [ [ [ [ [ [ , 3,] co-firing _3. subunit methyl] _5_thiathiazepine-bicyclic I ·2·〇]oct-2-ene-2-carboxylic acid dihydrochloride: 145413.doc 201028427 C(Ph)3 Reacts with 90% formic acid; separates the obtained solid; and uses a gradient of water and increasing acetonitrile concentration, w and 'error are purified by MCI gel chromatography; and collected from the suspension of the obtained (tetra) pure compound from the chromatography The polymorph; still removes acetonitrile by early precipitation and separates the obtained cefotaxol polymorph 145413.doc