CN1732167A - Monohydrate solvates of loracarbef - Google Patents

Abstract

This invention relates to monohydrate solvates of loracarbef. The invention also relates to processes for preparing solvates of loracarbef, crystalline monohydrate of loracarbef from said solvates and pharmaceutical compositions that include the crystalline monohydrate of loracarbef.

Description

Laura Kabi monohydrate solvate

field of invention

The present invention relates to the monohydrate solvate of loracarbef. The present invention also relates to processes for the preparation of solvates of loracarb, to processes for the preparation of crystalline monohydrate of loracarb from said solvates, and to pharmaceutical compositions comprising said crystalline monohydrate of loracarb way of things.

Background of the invention

Loracarbi is a synthetic beta-lactam antibiotic of carbacephems for oral administration. It has been disclosed in US Patent 4335211. Chemically, Loracarbyl is (6R,7S)-7-[(R)-2-amino-2-phenylacetamido]-3-chloro-8-oxo-1-azabicyclo( azabicyclo)[4.2.0]oct-2-ene-carboxylic acid monohydrate, which has the formula I.

Formula I

In laboratory tests, Loracarb was active against a broad spectrum of bacteria. Loracarbene has been shown to be the more stable compound with high blood levels and a long half-life.

Loracarbene can be isolated in various forms, including the crystalline monohydrate form, which has been disclosed in European Patent Publication EP0311366. The crystalline dihydrate form of Loracarbene has been disclosed in European Patent Publication EP0369686. Other known solvated forms of the compound are the di(DMF), dihydrate mono(DMF) and mono(DMF) forms which are disclosed in US Patent 4,977,257. US Patent 5580977 discloses a crystalline anhydrate of Lora Kabi.

The various solvates mentioned above are generally conventional intermediates for the preparation of loracarbidine, in particular the monohydrate form of loracarbidine. Compounds known to be useful in medicine need to have sufficient density for easy bulk product formulation. However, the process disclosed in EP0369686 yields loracarbyl monohydrate in the form of a fine, fluffy powder with a density of about 0.2 g/ml. This density makes batch formulation of Loracarbyl monohydrate very difficult.

Therefore, there is a great need for methods for the overall synthesis of desired compounds as well as intermediates for these compounds, particularly methods that can be adapted to large-scale manufacturing and result in high yields and reduced manufacturing costs

Summary of the invention

One aspect of the present invention provides a mono-N,N-dimethylacetamide monohydrate solvate of loracarbi represented by formula II-A.

Formula II-A

Another aspect of the present invention provides a mono-N-methylpyrrolidone monohydrate solvate of loracarbyl represented by formula II-B,

Formula II-B

Another aspect of the present invention provides a process for the preparation of the mono-N,N-dimethylacetamide monohydrate solvate and the mono-N-methylpyrrolidone monohydrate solvate of Loracarb.

Another aspect of the present invention provides a crystalline monohydrate of loracarb from mono-N,N-dimethylacetamide monohydrate solvate or mono-N-methylpyrrolidone monohydrate solvate of loracarb Methods.

Another aspect of the present invention provides a crystalline monohydrate of loracarbene having a bulk density greater than or equal to 0.6 gm/ml.

Another aspect of the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of crystalline monohydrate of loracarbene; and one or more pharmaceutically acceptable carriers, excipients or diluents, said loracarbene The bulk density of the crystalline monohydrate of Carby is greater than or equal to 0.6 gm/ml.

Detailed description of the invention

The present inventors have developed new monohydrate solvates of loracarb, specifically mono-N,N-dimethylacetamide monohydrate solvate represented by formula II-A and loracarb represented by formula II-B Mono-N-methylpyrrolidone monohydrate solvate.

Described single N, N-dimethylacetamide monohydrate solvate is characterized as follows with X-ray powder diffraction pattern: d I/I ₀ 15.6 17.0 11.80 100 11.12 41 7.43 25 5.91 12 5.19 14 4.88 16 4.76 twenty two 4.69 17 4.45 13 4.28 13 3.93 70 3.639 28 3.33 18 3.177 71 2.949 18 2.729 13 2.6122 13

The single N-methylpyrrolidone monohydrate solvate is characterized by X-ray powder diffraction as follows d I/I ₀ 15.8248 14 15.2251 13 12.0338 100 8.0594 8 7.5189 33 5.9968 13 5.4668 12 5.3810 14 5.2605 18 4.8863 twenty two 4.7513 37 4.4579 twenty one 4.2997 twenty two 4.1411 16 3.9939 55 3.6421 38 3.3858 18 2.7314 15

The above diffraction pattern was obtained with a Rigaku (RINT 2000) instrument for nickel-filtered copper radiation (Cu:Ni) at wavelength λ = 1.5406 Angstroms. The interplanar spacing is in the column marked "d", and the unit is Angstrom, and the relative intensity is in the column marked "I/I ₀ ".

The present inventors have also developed a process for the preparation of the mono-N,N-dimethylacetamide monohydrate solvate and the mono-N-methylpyrrolidone monohydrate solvate of Loracarb. The present inventors have also developed a method for the preparation of loracarbyl crystals represented by formula I from mono-N,N-dimethylacetamide monohydrate solvate and mono-N-methylpyrrolidone monohydrate solvate of loracarbene The monohydrate method. The resulting crystalline monohydrate of loracarbene has a bulk density greater than or equal to 0.6 gm/ml. The present inventors have also developed loracarbene crystalline monohydrate comprising bulk density greater than or equal to 0.6 gm/ml and admixed with one or more solid or liquid pharmaceutically acceptable diluents, carriers and/or excipients pharmaceutical composition.

In one aspect, the mono-N,N-dimethylacetamide monohydrate solvate of Loracarbi represented by Formula II-A is prepared by a process comprising mixing a compound represented by Formula III with N, N-dimethylacetamide and cyclic amine base or dimethylbenzylamine comprising 0-1 oxygen atoms form the free amine of the compound represented by formula IV,

Formula III

wherein R ₁ is hydrogen, trihalogen (C ₁ -C ₄ alkyl), C ₁ -C ₄ alkyl, C ₁ -C ₄ substituted alkyl, C ₁ -C ₄ alkoxy, C ₁ -C ₄ substituted Alkoxy, C ₁ -C ₆ alkylthio, C ₁ -C ₆ substituted alkylthio, methoxymethyl, carbamoyloxymethyl, acetoxymethyl, C ₂ -C ₆ alkenes group, C ₂ -C substituted _alkenyl , or halogen (such as bromine, chlorine, fluorine and iodine); R ₂ is a carboxyl protecting group,

Formula IV

reacting the free amine with an acylating agent represented by formula V,

Formula V

Wherein _R is an amino protecting group and L is a leaving group.

In another aspect, the mono-N-methylpyrrolidone monohydrate solvate of loracarbene represented by formula II-B is prepared by a process comprising:

Mixing the compound represented by formula III with N-methylpyrrolidone and a cyclic amine base or dimethylbenzylamine containing 0-1 oxygen atoms to form the free amine of the compound represented by formula IV,

Formula III

wherein R ₁ is hydrogen, trihalogen (C ₁ -C ₄ alkyl), C ₁ -C ₄ alkyl, C ₁ -C ₄ substituted alkyl, C ₁ -C ₄ alkoxy, C ₁ -C ₄ substituted Alkoxy, C ₁ -C ₆ alkylthio, C ₁ -C ₆ substituted alkylthio, methoxymethyl, carbamoyloxymethyl, acetoxymethyl, C ₂ -C ₆ alkenes group, C ₂ -C substituted _alkenyl , or halogen (such as bromine, chlorine, fluorine and iodine); R ₂ is a carboxyl protecting group,

Formula IV

reacting the free amine with an acylating agent represented by formula V,

Formula V

Wherein _R is an amino protecting group and L is a leaving group.

The term "carboxy protecting group" refers to an ester derivative of a carboxylic acid group which is not sterically hindered and which is generally used to block or protect the carboxylic acid group when the reaction is carried out on other functional groups of the compound group. Examples of such groups include allyl, alkyl, benzyl, substituted benzyl, silyl and haloalkyl groups such as 2,2,2-trichloroethyl, 2,2,2-tribromoethyl and 2-iodoethyl. Other examples of these groups include, for example, these groups, which can be found in E. Haslam, "Protecting Groups in Organic Chemistry", ed. J.G.W. McOmie, Plenum Press, New York, NY, 1973, Chapter 5 and T.W. Greene, "Protecting Groups in Organic Synthesis", John Wiley and Sons, New York, NY, 1981, Chapter 5. Specifically, the carboxyl protecting group is 4-nitrobenzyl.

The term "amino protecting group" refers to an amino substituent, typically used to block or protect an alkyl functionality when reactions are performed on other functional groups of the compound.

The amino protecting group R ₃ includes carbamate (eg tert-butoxycarbonyl or benzyloxycarbonyl) or enamine. In particular, the amino protecting groups include tert-butoxycarbonyl, phenoxyacetyl and enamines derived from (C ₁ -C ₄ alkyl)acetoacetate groups. Other amino protecting groups used in the field of cephalosporins, penicillins and peptides are also included in the above terms. Additional examples of groups referred to by the above terms can be found in JW Barton, "Protecting Groups in Organic Chemistry", ed. JGW McOmie, Plenum Press, New York, NY, 1973, Chapter 2 and TW Greene, "Protecting Groups in Organic Synthesis ", John Wiley and Sons, New York, NY, 1981, Chapter 7.

The term "leaving group" means a group which will leave under the reaction conditions, thereby attaching a free amine to the carbonyl. The leaving group includes the following groups, wherein L is a group represented by formula VI,

Formula VI

Wherein R ₄ is C ₁ -C ₆ alkyl, or L is Cl, Br, I, an active ester, such as p-nitrophenyl; or an adduct of dicyclohexylcarbodiimide.

The base includes compounds consisting of five- or six-membered tertiary cyclic amines which may contain oxygen atoms, or dimethylbenzylamine. Specifically, the tertiary cyclic amine bases include N-methylmorpholine (NMM) and N-methylpiperidine (NMP). The base may be used in an amount of about 1-1.3 molar equivalents, such as about 1.13 molar equivalents.

The hydrochloride salt of formula III can be prepared by the method described in European patent application 0266896.

In general, the amino and carboxyl protecting groups can be removed by methods known in the art. These examples include, for example, methods described in works on the subject, e.g. E. Haslam, "Protecting Groups in Organic Chemistry", ed. J.G.W. McOmie, Plenum Press, New York, NY, 1973, pp. 2 and 5 Zhang and T.W. Greene, "Protecting Groups in Organic Synthesis", John Wiley and Sons, NewYork, NY, 1981, Chapters 5 and 7.

To deprotect the protected amino group and the protected carboxyl group, a mixture of concentrated hydrochloric acid in water (2:1 ) is added to the acylation solution at a temperature of 0°C to -10°C within 30-45 minutes. Then, zinc powder (about 3.5 equivalents) was added over about 50-70 minutes, maintaining a temperature below 0°C. About 1.2 equiv of HCl was added and the reaction mixture was allowed to warm to ambient temperature over 45-60 minutes. This mixture was stirred at ambient temperature for about 5-6 hours, semicarbazide hydrochloride (1.15 equiv) was added, followed by stirring for 30-60 minutes. The pH was adjusted to about 2.9-3.1 with 28% aqueous ammonia, and the mixture was filtered through a celite bed. The filtrate was heated to 48-55°C, and its pH was adjusted to 4.8-5.0 with 28% aqueous ammonia. The separated solid was stirred for an additional 30 minutes and the pH was continuously adjusted to 5.8-6.2. The temperature of the mixture was lowered to 20-25° C., and a polar solvent such as acetone was added and stirred for another 30 minutes. The crystals were collected by filtration, washed with acetone, cooled to 20-25°C, and dried to obtain mono-N,N-dimethylacetamide monohydrate solvate of loracarbi or mono-N-methylpyrrolidone monohydrate compounds.

In another aspect, the mono-N,N-dimethylacetamide monohydrate solvate of loracarb and the mono-N-methylpyrrolidone monohydrate solvate can be converted to the crystalline monohydrate of loracarb. It has been found that loracarbidine monohydrate prepared from mono-N,N-dimethylacetamide monohydrate solvate or mono-N-methylpyrrolidone monohydrate has a concentration equal to or greater than 0.6 g/ Bulk density in ml.

Typically, the monohydrate of loracarbene is prepared by suspending the mono-N,N-dimethylacetamide monohydrate solvate of loracarbene or the mono-N-methylpyrrolidone monohydrate solvate of loracarbene in water. A clear solution of the material can be obtained by adding a minimal amount of acid, usually 6N (or more dilute) hydrochloric acid. The temperature of the solution was raised to about 50°C, and then 28% ammonia solution was slowly added to the solution until a pH of about 4.8 was obtained. During the addition of the base, the developing suspension was stirred and kept at about 50°C. The warm pH-adjusted suspension (50°C) was cooled to about 20°C, stirred, filtered, and the collected solids were dried at 40-45°C to give crystalline Laura Carbyl monohydrate.

The obtained crystalline monohydrate of loracarbene can be formulated into conventional dosage forms, such as tablets, capsules, pills, solutions, etc., and the bulk density of the crystalline monohydrate of loracarbene is equal to or greater than 0.6 g/ml. In these cases, the medicament can be prepared by conventional methods using conventional pharmaceutical excipients.

The present invention can also be further illustrated by the following examples, which are merely examples of the present invention and do not limit the scope of the present invention. Although the examples refer to mono-N,N-dimethylacetamide monohydrate solvate of loracarb and mono-N-methylpyrrolidone monohydrate solvate, crystalline monohydrate of loracarb, these implementations The principles in this example can be applied to other solvates of Lora Kabi.

In the following examples, the terms N,N-dimethylacetamide monohydrate solvate of Loracarb, nuclear magnetic resonance spectrum, mass spectrum and infrared spectrum are abbreviated as N,N-DMAc, NMR, MS and IR, respectively. In NMR spectra, the following abbreviations are used: "s" is singlet, "d" is doublet, "t" is triplet, "q" is quadruplet, "m" is multimodal.

The NMR spectra were obtained from a Bruker (DRX 300) 300 MHz instrument. Chemical shifts are expressed in ppm (parts per million downfield from tetramethylsilane) values.

Example 1

Preparation of mono-N,N-dimethylacetamide solvate of Loracarbi

Step A: Preparation of N-Methylmorpholine Salt

At 20-25°C, in a mixture of N,N-dimethylacetamide (60ml) and N-methylmorpholine (3.0g), add p-nitrobenzyl 7β-amino-3-chloro-hydrochloride in portions 1-carba (1-dethia)-3-cephem-4-carboxylic acid (10.0 g) to form the free amine. The reaction mixture was stirred for 30 minutes, then cooled to -5 to -10°C.

Step B: Preparation of Mixed Anhydride

9.3 g of the Na/K Dane salt of phenylglycine (prepared according to Dane et al., Angew.Chem, Vol. 74, p. 873, 1962) were suspended in N,N-dimethylacetamide (150 ml), and Stir for 30-40 minutes. The reaction mixture was cooled to -20 to -15°C, and methanesulfonic acid (0.12 g) and N-methylmorpholine (0.06 g) were added thereto. Ethyl chloroformate (3.3 g) was added in one portion and stirring was continued at -10 to -15°C for 90 minutes.

Step C: Condensation

Slowly add the N-methylmorpholine hydrochloride solution from step A containing free amines to the mixed anhydride from step B at -20 to -10°C. The reaction mixture was stirred for 2 hours and the progress of the reaction was monitored by TLC or HPLC. After completion of the reaction, a mixture of concentrated hydrochloric acid in water (28 ml concentrated hydrochloric acid in 14 ml H ₂ O) was added over a period of 10-15 minutes to double protected (diprotected) loracabie followed by zinc powder (6.0 g) and kept Said temperature is less than +5°C. The temperature was raised to 20-25°C and the reaction mixture was stirred for about 2 hours. Semicarbazide hydrochloride (3.3 g) was added and stirring was continued for 30-35 minutes. The pH of the reaction mixture was adjusted to 2.9-3.0 using 28% ammonia solution and then filtered. The filtrate is heated to about 48-55°C and the pH is adjusted to 4.8-5.0. The separated solid was stirred for an additional 30 minutes and finally the pH was adjusted to 5.8-6.2. The reaction mixture was cooled to 20-25°C, acetone was added, and stirred for an additional 30 minutes. It was then filtered and washed with acetone. The solid was dried in vacuo at 40-42°C to give loracarbi mono-N,N-DMAc monohydrate solvate, which was characterized based on the following data:

Dry weight 9.0g

Yield w/w 0.9

NMR (D ₂ O-DCl) (300MHz): 7.44-7.45 (s, 5 H , Ar H ), 5.35 (d, 1 H -β-lactam), 5.2 (s, 1 H , CH -phenyl ), 3.93-(m, 1 H -β-lactam) 2.91-3.03 (s, s, 6 H , N(CH ₃ ) ₂ ) 2.55 (m, 2 H , CH ₂ ), 2.05 (s, 3H, COCH ₃ ) 1.63 (m, 1 H , CH) 1.31 (m, 1 H , CH)

Water content (according to KF) = 3.0%

IR (KBr sheet) = 2980-3660 (s, and width) 1780, 1700, 1630, 1580, 1460, 1400, 1390, 1380 (m to strong).

Example 2

Preparation of crystalline monohydrate of loracarb from mono-N,N-DMAc monohydrate solvate

Loracabie's mono-N,N-dimethylacetamide monohydrate solvate (10.0 g) was suspended in water (80 ml). 12N hydrochloric acid (1.0 ml) was added to obtain a clear solution. Activated carbon (1.0 g) was added and the reaction mixture was stirred for 30-40 minutes. The suspension was then filtered and washed with water (30ml). The temperature of the filtrate was raised to 50-55 °C, and the pH was slowly adjusted to 1.8-1.9 with 8% _NH3 solution. The reaction mixture was stirred at 50-55°C for 30 minutes. Stirring was continued for another 30 minutes, then slowly cooled to 20-25°C. The slurry was washed with water. The filter cake was dried in an air oven at 40-45°C to give crystalline loracarbene monohydrate (5.0 g) with a bulk density greater than 0.6 g/ml. The IR, NMR and X-ray diffraction patterns of the crystalline loracarbene monohydrate were consistent with authentic samples of crystalline loracarbene monohydrate.

Example 3

Preparation of Mono-N-Methylpyrrolidone Monohydrate Solvate of Loracarbi

Step A: Preparation of N-methylmorpholine salt:

To a mixture of N-methylpyrrolidone (60ml) and N-methylmorpholine (3.0g) was added p-nitrobenzyl 7βamino-3-chloro- 1-carba (1-dethia)-3-cephem-4-carboxylic acid (10.0 g). The reaction mixture was stirred for 60 minutes.

Step B: Preparation of Mixed Anhydride

9.5 g of Na/K Dane salt (prepared according to Dane et al., Angew. Chem., Vol. 74, p. 873, 1962) were suspended in N-methylpyrrolidone (120 ml) and stirred for 30-35 minutes. The reaction mixture was cooled to -20 to -15°C, and methanesulfonic acid (0.15 g) and N-methylmorpholine (0.08 g) were added thereto. Ethyl chloroformate (3.3 g) was added in one portion and stirring was continued at -10 to -15°C for 60-90 minutes.

Step C: Condensation

Slowly add the N-methylmorpholine hydrochloride solution obtained from step A containing free amines to the mixed anhydride obtained from step B at -20 to -10°C in about 15-20 minutes. The reaction mixture was stirred for 60 minutes. At -10 to 0°C, a solution of concentrated hydrochloric acid in water (28ml of concentrated hydrochloric acid in 14ml of H ₂ O) was added dropwise to the double-protected Loracarb, and then zinc powder (6.0g) was added while the temperature was Keep at 0 to +5°C. The temperature was raised to 20-25°C and the reaction mixture was stirred for about 60 minutes. Semicarbazide hydrochloride (3.3 g) was added and stirring continued for 30 minutes. The pH of the reaction mixture was adjusted to 2.9-3.0 with 28% _NH3 solution, then filtered. The filtrate was washed with N-methylpyrrolidone (50ml), and the pH was adjusted to 4.8-5.0. The separated solid was stirred for about another 30 minutes and finally the pH was adjusted to 5.8-6.2. The reaction mixture was cooled to 20-25°C, acetonitrile (60ml) was added and stirred for a further 30 minutes. It was then filtered and the solid was dried in vacuo to afford the mono-N-methylpyrrolidone monohydrate solvate of loracarbene, which was characterized based on the following data:

NMR (300MHz) (s): 7.4 (s, 5 H , Ar H ), 5.3 (d, 1 H , β-lactam), 5.2 (s, 1 H , CH, phenyl), 3.83 (m, 1 H , β-lactam), 3.3-3.42 (t, 2 H , derived from N-methylpyrrolidone), 2.72 (s, 3 H , N-CH ₃ , derived from NMP), 2.46-2.53 (m, 2 H , CH ₂ ), 2.32-2.37 (t, 2 H , from NMP), 1.90-1.95 (m, 2 H , from NMP), 1.55 (m, 1 H , CH), 1.18-1.22 (m, 1 H , CH)

Moisture content (according to KF): 5.0% by weight

IR (KBr sheet): 2980-3650 (s, and wide) 1780, 1720, 1690, 1600, 1580, 1460, 1400.

Example 4

Synthesis of crystalline monohydrate of loracarb from mono-N-methylpyrrolidone monohydrate solvate

Loracarbyl mono-N-methylpyrrolidone monohydrate solvate (10.0 g) was suspended in water (80 ml). 12N hydrochloric acid (1.0 ml) was added to obtain a clear solution. Activated carbon (1.0 g) was added and the reaction mixture was stirred for 30-40 minutes. The suspension was then filtered and washed with water (30ml). The temperature of the filtrate was increased to 50-55°C, and the pH was slowly adjusted to 1.8-1.9 with 8% ammonia solution. The reaction mixture was stirred at 50-55°C for 30 minutes, and the pH was slowly adjusted to 4.5-4.8 at 50-55°C with stirring within 30-35 minutes. Stirring was continued for another 30 minutes, then slowly cooled to 20-25°C. The slurry was washed with water. The filter cake was dried in an air oven at 40-45°C to give crystalline loracarbene monohydrate (5.0 g) with a bulk density greater than 0.6 g/ml.

The IR, NMR and X-ray diffraction patterns of the crystalline loracarbene monohydrate were consistent with authentic samples of crystalline loracarbene monohydrate.

While this invention has been described in terms of specific examples thereof, certain modifications and equivalents will be apparent to those of ordinary skill in the art and are intended to be encompassed within the scope of this invention.

Claims (20)

1. single N,N-dimethylacetamide one monohydrate solvates of the Laura card ratio represented of formula II-A,

Formula II-A.

2. compound as claimed in claim 1 is characterized in that it has following X-ray powder diffraction collection of illustrative plates:

d I/I ₀

15.6 17.0

11.80 100

11.12 41

7.43 25

5.91 12

5.19 14

4.88 16

4.76 22

4.69 17

4.45 13

4.28 13

3.93 70

3.639 28

3.33 18

3.177 71

2.949 18

2.729 13

2.6122 13。

3. single N-Methyl pyrrolidone one monohydrate solvates of the Laura card ratio represented of formula II-B

Formula II-B.

4. compound as claimed in claim 3 is characterized in that it has following X-ray powder diffraction collection of illustrative plates:

d I/I ₀

15.8248 14

15.2251 13

12.0338 100

8.0954 8

7.5189 33

5.9968 13

5.4668 12

5.3810 14

5.2605 18

4.8863 22

4.7513 37

4.4579 21

4.2997 22

4.1411 16

3.9939 55

3.6421 38

3.3858 18

2.7314 15。

5. the method for single N,N-dimethylacetamide one monohydrate solvates of the Laura card ratio represented of a preparation formula II-A,

Formula II-A

Described method comprises compound and N,N-dimethylacetamide and the alkali that mixes formula III, the unhindered amina of the compound that the formula IV of formation represents,

Formula III

R wherein ₁Be hydrogen, three halogen (C ₁-C ₄Alkyl), C ₁-C ₄Alkyl, C ₁-C ₄Substituted alkyl, C ₁-C ₄Alkoxyl group, C ₁-C ₄Substituted alkoxy, C ₁-C ₆Alkylthio, C ₁-C ₆Substituted alkane sulphur base, methoxymethyl, carbamoyloxy methyl, acetoxy-methyl, C ₂-C ₆Alkenyl, C ₂-C ₆Substituted alkenyl base or halogen, for example bromine, chlorine, fluorine and iodine; R ₂Be carboxy protective group,

Formula IV

The acylting agent reaction that compound that the formula IV of making represents and formula V represent

Formula V

R wherein ₃Be the amido protecting group, L is a leavings group.

6. the method for single N-Methyl pyrrolidone one monohydrate solvates of the Laura card ratio represented of a preparation formula II-B,

Formula II-B

Described method comprises mixes compound and N-Methyl pyrrolidone and the alkali that formula III is represented, the unhindered amina of the compound that the formula IV of formation represents,

Formula III

R wherein ₁Be hydrogen, three halogen (C ₁-C ₄Alkyl), C ₁-C ₄Alkyl, C ₁-C ₄Substituted alkyl, C ₁-C ₄Alkoxyl group, C ₁-C ₄Substituted alkoxy, C ₁-C ₆Alkylthio, C ₁-C ₆Substituted alkane sulphur base, methoxymethyl, carbamoyloxy methyl, acetoxy-methyl, C ₂-C ₆Alkenyl, C ₂-C ₆Substituted alkenyl base or halogen, for example bromine, chlorine, fluorine and iodine; R ₂Be carboxy protective group,

Formula IV

The acylting agent reaction that compound that the formula IV of making represents and formula V represent

Formula V

R wherein ₃Be the amido protecting group, L is a leavings group.

7. as claim 5 or 6 described methods, it is characterized in that described carboxy protective group R ₂Be selected from base, alkyl, benzyl, substituted benzyl, silyl, haloalkyl and alkoxyalkyl in the alkene.

8. method as claimed in claim 7 is characterized in that described carboxy protective group is the 4-nitrobenzyl.

9. as claim 5 or 6 described methods, it is characterized in that described amido protecting group R ₃Be selected from alkoxyl group-carbonyl, phenoxy group-carbonyl, phenoxy group-acyl group, alkoxyl group acyl group, aralkoxy-carbonyl, by C _1-4Alkyl acetylacetic ester deutero-enamino (enamino) and acyl group.

10. as claim 5 or 6 described methods, it is characterized in that described leavings group L is the compound that formula VI represents,

Formula VI

R wherein ₄Be selected from halogen such as chlorine, bromine, iodine or C _1-6The affixture of alkyl, benzyl, substituted benzyl, phenyl, substituted-phenyl, dicyclohexylcarbodiimide and alkoxyalkyl.

11., it is characterized in that described alkali is the cyclammonium alkali that comprises 0-1 Sauerstoffatom as claim 5 or 6 described methods.

12. method as claimed in claim 11 is characterized in that described cyclammonium alkali is selected from five yuan or hexa-atomic uncle's cyclammonium.

13. method as claimed in claim 12 is characterized in that described cyclammonium alkali is N-methylmorpholine or N methyl piperazine.

14. a method for preparing the crystallization monohydrate of Laura card ratio, described method comprises:

With single N,N-dimethylacetamide one monohydrate solvates of acid treatment Laura card ratio and

Regulate pH with alkali, to obtain the crystallization monohydrate of Laura card ratio.

15. a method for preparing the crystallization monohydrate of Laura card ratio, described method comprises:

With single N-Methyl pyrrolidone one monohydrate solvates of acid treatment Laura card ratio and

Regulate pH with alkali, to obtain the crystallization monohydrate of Laura card ratio.

16., it is characterized in that used acid is mineral acid or organic acid as claim 14 or 15 described methods.

17. method as claimed in claim 16 is characterized in that described acid is hydrochloric acid.

18., it is characterized in that used alkali is ammonia as claim 14 or 15 described methods.

19. the crystallization monohydrate of a Laura card ratio, its tap density is more than or equal to 0.6g/ml.

20. a pharmaceutical composition, it comprises:

The Laura card of treatment significant quantity is than crystallization monohydrate, described Laura card than the tap density of crystallization monohydrate more than or equal to 0.6g/ml;

And one or more pharmaceutically acceptable carriers, vehicle or thinner.