CN103298813A

CN103298813A - Amorphous powder and polymorph of carbapenem derivative, preparation method therefor and use thereof

Info

Publication number: CN103298813A
Application number: CN2011800612712A
Authority: CN
Inventors: 李翱; 崔永铭; 董岩岩
Original assignee: ZHUHAI EBANG PHARMACEUTICAL CO LTD; Xuanzhu Pharma Co Ltd
Current assignee: ZHUHAI YIBANG PHARMACEUTICAL CO Ltd; Shandong Xuanzhu Pharma Co Ltd
Priority date: 2010-12-16
Filing date: 2011-12-16
Publication date: 2013-09-11
Anticipated expiration: 2031-12-16
Also published as: WO2012079289A1; CN103298813B

Abstract

The present invention relates to an amorphous powder, a solvate, and a polymorph of a carbapenem derivative (4R,55,6S)3-[(3S,5S)-5-[(furan-2-ylmethyl)amino-formyl]3-pyrrolidine]thio-6-[(R)-1-hydroxyethyl]4-methyl-7-oxo-1-oxabicyclo[3.2.0]hept-2-enyl-2-carboxylic acid, that is, the compound of Formula (1), a preparation method therefor, a pharmaceutical composition containing the amorphous powder and/or the polymorph, and a use thereof in treatment and/or prevention of infectious diseases and in preparation of drugs for treating and/or preventing infectious diseases.

Description

Amorphous powder and polymorph of carbapenem derivative, and preparation method and application thereof

The present invention relates to amorphous powders, solvates and polymorphs of the carbapenem derivative (4R,5S,6S) -3- [ (3S,5S) -5- [ (furan-2-yl -yl) amine acyl ] -3-pyrrolidine ] thio-6- [ (R) -l-hydroxyethyl ] -4- yl-7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylic acid, processes for their preparation, pharmaceutical compositions containing the amorphous powders, solvates and/or polymorphs and their use. Background

Carbapenem antibiotics are a β -lactam antibiotic developed in the seventies, and are concerned because of their broad antibacterial spectrum, strong antibacterial activity, and stability to β -lactamase.

Due to abuse of antibiotics, clinical drug-resistant bacteria are increasing, and development of novel carbapenem antibiotics with high antibacterial activity and long half-life to various common clinical infection pathogenic bacteria in hospitals becomes a research hotspot of antibiotics.

WO 2009/000210A 1 discloses a class of carbapenem derivatives having high stability against β -lactamase and DHP-I, useful in β -lactamase-producing strains, which have a broad antibacterial spectrum, good antibacterial activity against gram-positive and negative, aerobic and anaerobic bacteria and nosocomial pathogenic bacteria, and a long half-life and long-lasting antibacterial action, example 1 of the PCT application describes the preparation of (4R,5S,6S) -3- [ (3S,5S) -5- [ (furan-2-yl yl) carbamoyl ] -3-pyrrolidine ] thio-6- [ (R) -l-hydroxyethyl ] -4- yl-7-oxomini-azabicyclo [3.2.0] hept-2-ene-2-carboxylic acid (compound A) which has a strong antibacterial activity against gram-positive, gram-negative and anaerobic bacteria, and this application is incorporated herein in its entirety.

The crystal form has certain influence on the physicochemical properties of the compound. The same pharmaceutical compound in different crystal forms may have significant differences in solubility, melting point, density, hardness, stability, hygroscopicity, appearance, etc., thereby affecting the stability of the drug, etc.

Disclosure of Invention

The present invention provides a compound represented by the following formula (1)

Namely amorphous powders, solvates and polymorphs of (4R,5S,6S) -3- [ (3S,5S) -5- [ (furan-2-yl yl) amine acyl ] -3-pyrrolidine ] thio-6- [ (R) -l-hydroxyethyl ] -4- yl-7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylic acid: form II, form III, form IV and form V, processes for their preparation, pharmaceutical compositions comprising the amorphous powders and/or polymorphs and their use for the preparation of a medicament for the treatment and/or prevention of infectious diseases and for the treatment and/or prevention of infections

The application of the medicine in treating diseases. In one aspect of the present invention, there is provided a compound represented by the following formula (1):

in the form of an amorphous powder.

In another aspect of the invention, the following embodiments are provided:

1. a compound of the following formula (1) in a crystalline form

This crystalline form is designated as form II crystal, form II, and is characterized by an X-ray powder diffraction pattern using Cu-Ka radiation, expressed at an angle of 2 Θ, having characteristic peaks at 6.5 deg.0.2, 11.0 ± 0.2, 16.8 deg.0.2, 23.0 deg.0.2.

2. Form II according to embodiment 1, characterized by an X-ray powder diffraction pattern using Cu-Ka radiation, expressed at an angle of 2 Θ, further having characteristic peaks at 15.1 st 0.2, 15.7 st 0.2.

3. Form II according to embodiment 2, characterized by an X-ray powder diffraction pattern using Cu-Ka radiation expressed in degrees 2 Θ further having characteristic peaks at 13.3 deg. 0,2, 21.1 ± 0.2, 22.1 ± 0.2, 24.6 ± 0.2.

4. The crystalline form II of any of embodiments 1-3, characterized by an X-ray powder diffraction pattern substantially as shown in figure 1.

5. Crystalline form II according to any of embodiments 1 to 4, characterized by a DSC curve with a maximum decomposition temperature of 165-185 ℃ or as shown in FIG. 2.

In another aspect of the present invention, the following embodiments are provided:

6. a compound of the following formula (1) in crystalline form

This crystalline form is designated as form III crystal, form III, and is characterized by an X-ray powder diffraction pattern using Cu-Ka radiation at an angle of 2 Θ having characteristic peaks at 5.3 + -0.2, 10.7 + -0.2, 15.7 + -0.2, 21.3 Shi 0.2.

7. Form III as claimed in embodiment 6, characterized by the use of Cu-Ka radiation

The X-ray powder diffraction pattern expressed by the angle of 2 theta also has characteristic peaks at 23.1 +/-0.2 and 27.3 +/-0.2.

8. Form III according to embodiment 7, characterized by an X-ray powder diffraction pattern using Cu-Ka radiation expressed in degrees 2 Θ further having characteristic peaks at 20.7 ± 0.2, 23.5 ± 0.2, 27.0 ± 0.2, 28.8 sec 0.2.

9. The crystalline form III of any one of embodiments 6-8, characterized by an X-ray powder diffraction pattern substantially as shown in figure 4.

In yet another aspect of the invention, the following embodiments are provided:

10. a compound represented by the following formula (1) in a crystalline form

The molecular structure of the crystal contains 1 molecule of water and 1 molecule of alcohol, is a solvate, and has a structure shown in the formula

This crystalline form is designated as form IV crystal, form IV, and is characterized by an X-ray powder diffraction pattern using Cu-Ka radiation at an angle of 2 Θ having characteristic peaks at 6.5 deg.C 0.2, 10.9 deg.C 0.2, 12.9 + -0.2, 26.1 + -0.2.

11. Form IV, as described in embodiment 10, characterized by an X-ray powder diffraction pattern using Cu-Ka radiation expressed in degrees 2 Θ further having characteristic peaks at 39.6 deg.0.2, 32.8 ± 0.2.

12. Form IV according to embodiment 11, characterized by an X-ray powder diffraction pattern using Cu-Ka radiation expressed in degrees 2 Θ further having characteristic peaks at 21.2 deg.0, 2, 22.0 ± 0.2, 22.8 ± 0.2, 24.4 ± 0.2.

13. Form IV according to any one of embodiments 10 to 12, characterized by an X-ray powder diffraction pattern substantially as shown in figure 6.

In yet another aspect of the invention, the following embodiments are provided:

14. a compound represented by the formula (1) in a crystalline form

This crystalline form is designated as form V crystal, form V, and is characterized by an X-ray powder diffraction pattern using Cu-Ka radiation at an angle of 2 Θ having characteristic peaks at 6.1 + -0.2, 10.3 + -0.2, 11.6 Shi 0.2, 20.8 + -0.2. 15. Form V according to embodiment 14, characterized by an X-ray powder diffraction pattern using Cu-Ka radiation expressed at an angle of 2 Θ further having characteristic peaks at 16.5 ± 0.2, 21, 5X 0.2.

16. Form V according to embodiment 15, characterized by an X-ray powder diffraction pattern using Cu-Ka radiation expressed in degrees 2 Θ further having characteristic peaks at 9.0 ± 0.2, 19.5 ± 0.2, 19.8 ± 0.2, 22.2 Shi 0.2.

17. The crystalline form V of any one of embodiments 14-16, characterized by an X-ray powder diffraction pattern substantially as shown in figure Ί.

18. Form V according to any one of embodiments 14 to 17, characterized by a DSC curve as shown in figure 8.

Comprises the crystalline form II according to any one of embodiments 1 to 5 and the crystalline form V according to any one of embodiments 14 to 18.

The present invention also provides amorphous powders, solvates, and polymorphs of a compound represented by formula (1): preparation methods of crystal form II, crystal form III, crystal form IV and crystal form V. Preparation of amorphous powder of Compound of formula (1)

Amorphous powders of compounds of formula (1) may be obtained by a process comprising the steps of:

wherein PG_rX represents an amino protecting agent, PG₂X represents a carboxyl protecting agent, AA-X represents a hydroxyl activating agent, wherein PGi represents an amino protecting group, PG₂Represents a carboxyl protecting group, AA represents a leaving group, X represents a halogen selected from fluorine, chlorine, bromine and iodine, preferably chlorine; CH (CH)₃CO-S-M represents an alkali metal thioacetate, wherein M represents an alkali metal cation such as potassium ion, sodium ion, etc.;

performing amino protection reaction on L-hydroxyproline and an amino protection reagent PG X to form an intermediate b, further reacting with ^ in the presence of a coupling activator to form an intermediate c, reacting the intermediate c with a hydroxyl activator AA-X to form an intermediate d, reacting with alkali metal thioacetate to generate an intermediate e, further performing deprotection reaction by alkali hydrolysis, and removing CH₃The CO-moiety forms an intermediate f with f under basic conditionsThe reaction forms intermediate g, which is then deprotected to yield the compound of formula (1).

The "amino protection reaction" refers to a conventional reaction in which an active hydrogen on an amino group in a compound is protected with an amino protecting agent, which may be a conventional amino protecting agent such as PG_rX, may also be other amino group protecting groups commonly used by those skilled in the artReagents for protecting the reagents; wherein PGi is selected from, for example, but not limited to, p-nitrobenzyloxycarbonyl (PNZ), benzyloxycarbonyl (Cbz), t-butoxycarbonyl (Boc), fluorene oxycarbonyl (Fmoc), allyloxycarbonyl (Alloc), tris silicon ethoxycarbonyl (Teoc), oxycarbonyl, ethoxycarbonyl, o-phenyl-bis acyl (Pht), p-toluenesulfonyl (Tos), trifluoroacetyl (Tfa), p-nitrobenzenesulfonyl (Ns), pivaloyl, benzene acyl, triphenyl (Trt), 2, 4-bis oxybenzyl (Dmb), p- oxybenzyl (PMB), etc., preference is given to p-nitrobenzyloxycarbonyl (PNZ), benzyloxycarbonyl (Cbz), tert-butoxycarbonyl (Boc), p- oxybenzyl (PMB).

The carboxyl protection reaction is a reaction for protecting active hydrogen on carboxyl in a compound by using a carboxyl protection reagent, and the carboxyl protection reagent can be a reagent used by a conventional carboxyl protection reagent; wherein PG₂Selected from, for example, but not limited to, p-nitrobenzyl (PNB), yl, methoxy yl, thio yl, tetrahydropyranyl, tetrahydrofuranyl, oxyethyl yl, allyl, benzyloxy yl, benzene acyl yl, p-bromobenzene acyl yl, (X- phenylbenzene acyl yl, p- oxybenzene acyl yl, diacylmethyl, N-phthalimido yl, ethyl, 2,2, 2-trichloroethyl, 2-methylthioethyl, t-butyl, benzyl, 2, 4, 6-tri ylbenzyl, p-bromobenzyl, o-nitrobenzyl, p- oxybenzyl, 4-pyridine yl, tri yl silyl, triethylsilyl, t-butyl di yl silyl, diphenyl group, Ν' -diisopropylhydrazide and the like, p-nitrobenzyl (Ν β), benzyloxy group, tert-butyl group, benzyl group, p-bromobenzyl group, o-nitrobenzyl group, p- oxybenzyl group, p-bromobenzyl group, diphenyl group are preferable. The "coupling activator" refers to a conventional reagent that causes the condensation reaction between the carboxyl group of intermediate b and the amino group in ^ and is selected from, for example, but not limited to, [ isopropyl derivative acid, -yl sulfonyl chloride, p- benzenesulfonyl chloride, isobutyl derivative acid, N-Carbonyl-diimidazole,' tris -ylacetyl chloride and the like, preferably isopropyl formate, Ν -carbonyl-diimidazole; wherein "halo" means fluoro, chloro, bromo or iodo, preferably chloro, for example chloro isopropyl acetate.

The "hydroxyl activation reaction" refers to a conventional activation reaction in which active hydrogen on hydroxyl in intermediate C is activated into leaving group ALPHA-by using conventional hydroxyl activator, so that alkali metal thioacetate can better perform nucleophilic substitution reaction to prepare intermediate e, wherein the hydroxyl activator AA-X is selected from, for example, but not limited to, methylsulfonyl chloride, halogenated isopropyl ester, etc.; wherein the "halo" refers to fluoro, chloro, bromo or iodo, preferably chloro, etc.

The "deprotection reaction" refers to a reaction of removing a carboxyl protecting group, an amino protecting group and other protecting groups by a conventional method, and during the deprotection of the groups, catalysts such as nickel catalysts (Raney Ni, supported nickel, reduced nickel and nickel boride), palladium catalysts (palladium oxide,: palladium black and supported palladium), platinum catalysts (platinum oxide, platinum black and supported platinum), preferably palladium catalysts such as palladium/carbon and palladium chloride; metal hydrides such as lithium aluminum hydride, diborane, sodium borohydride and the like may also be used as catalysts.

In carrying out the above reaction, a basic or acidic environment can be produced by selecting an appropriate base or acid depending on the kind of the protecting agent used, the base including organic bases and inorganic bases, and the acid including organic acids and inorganic acids.

The inorganic base is selected from, for example, but not limited to, potassium hydroxide, sodium hydroxide, zinc hydroxide, calcium hydroxide, potassium carbonate, potassium bicarbonate, sodium carbonate, sodium bicarbonate, and the like.

The organic base is selected from, for example, but not limited to, amine compounds such as amine, tris amine, diethylamine, triethylamine, diisopropylamine, ethylenediamine, triethanolamine, diisopropylethylamine, tributylamine, N-bis phenylamine, dicyclohexylamine, dibenzylamine, N-benzyl- β -phenylethylamine, 1-diphenylhydroxy amine, N, -dibenzylethylenediamine, diethanolamine, triethanolamine, dimethylethanolamine, 2- (diethylamino) ethanol, 2-aminoethanol, tromethamine, and mixtures thereof,

Ν - based glucosamine, etc.; basic amino acids such as l-arginine, lysine, histidine, etc.; quaternary ammonium bases such as betaine, choline, tris hydroxy ethyl ammonium, and the like; alkaloids such as procaine, caffeine, ephedrine, etc.; alkali metal salts of alcohols such as sodium methoxide, potassium acetate, potassium tert-butoxide, etc.; alkylmetal lithium compounds such as butyl lithium, phenyl lithium and the like; lithium amido compounds such as Lithium Diisopropylamide (LDA), lithium hexa -disilazane (LiHMDS), and the like; other basic substances, such as 1- (2-hydroxyethyl) pyrrole, 4- (2-hydroxyethyl) morpholine, N- -ylmorpholine, imidazole, pyridine, N- -ylpiperidine and the like.

The inorganic acid is selected from, for example, but not limited to, hydrobromic acid, hydrochloric acid, sulfuric acid, selenite, nitric acid, phosphoric acid, and the like.

The organic acid is selected from, for example, but not limited to, sulfonic acid, dodecylsulfuric acid, 2-naphthalenesulfonic acid, benzenesulfonic acid, oxalic acid, 2-dichloroacetic acid, glycerophosphoric acid, 2-hydroxyethanesulfonic acid, L-aspartic acid, maleic acid, ethanesulfonic acid, 1, 5-disulfonic acid naphthalene, ethane-1, 2-disulfonic acid, cyclohexylsulfamic acid, p-toluenesulfonic acid, and the like. Preparation of polymorphic forms of the compound of formula (1)

In one aspect of the present invention, there is provided a process for preparing a crystalline form II of the compound of formula (1): the first scheme is as follows: adding alkali into the water suspension of amorphous powder of the compound shown in the formula (1) until the compound is completely dissolved, adjusting the obtained solution to pH <7 with acid, for example, pH value is about 5-6, precipitating crystals, filtering, and drying to obtain the compound shown in the formula (1) crystal form II.

Scheme II: adding a base to the aqueous suspension of the amorphous powder of the compound of formula (1) until the compound is completely dissolved, adding a good solvent, adjusting the resulting mixture with an acid to a pH of less than 7, e.g. a pH of about 5-6, precipitating crystals, filtering and drying to obtain the compound of formula (1) in crystalline form II.

The third scheme is as follows: adding a good solvent into the water suspension of the amorphous powder of the compound shown in the formula (1) until the compound is completely dissolved, decompressing and distilling the good solvent, filtering, precipitating crystals, and drying to obtain the crystal form II of the compound shown in the formula (1).

And the scheme is as follows: and (2) adding a good solvent into the water suspension of the amorphous powder of the compound shown in the formula (1) until the compound is completely dissolved, adding a poor solvent, separating out crystals, filtering and drying to obtain the crystal form II of the compound shown in the formula (1).

In the above-mentioned scheme two, scheme three and scheme four, the good solvent refers to a solvent in which the compound of formula (1) is soluble, and is selected from, for example, but not limited to, alcohol, ethanol, n-propanol, acetonitrile, tetrahydrofuran or a mixture thereof, and a water- alcohol mixture, a water-ethanol mixture, and the like, preferably alcohol-water solution or ethanol-water solution or tetrahydrofuran at a volume ratio of 5 to 95%.

In the fourth embodiment, the poor solvent is a solvent in which the compound of formula (1) is insoluble or slightly soluble, and is selected from, for example, but not limited to, isopropanol.

And a fifth scheme: and (2) heating the water suspension of the amorphous powder of the compound shown in the formula (1) until the compound is completely dissolved, cooling to separate out crystals, filtering, and drying to obtain the crystal form II of the compound shown in the formula (1).

In another aspect of the present invention, there is provided a process for preparing form III of the compound of formula (1): heating and stirring a water/acetonitrile (such as 4: 3) suspension of amorphous powder of the compound shown in the formula (1) until the compound is dissolved, filtering, freezing and crystallizing the filtrate, and filtering to obtain the compound shown in the formula (1) in the crystal form III.

In yet another aspect of the present invention, there is provided a process for preparing form IV of the compound of formula (1): adding alkali into the water suspension of the amorphous powder of the compound shown in the formula (1) until the compound is completely dissolved, adding a polar organic solvent methanol, adjusting the pH value of the obtained mixture to be less than 7 by using acid, such as 5-6, and then, placing at low temperature of 0-5 ℃ for crystallization to obtain the crystal form IV of the compound shown in the formula (1).

In yet another aspect of the present invention, there is provided a process for preparing form V of the compound of formula (1): general formula

(1) Stirring compound crystal form II in a mixture of water and a water-miscible organic solvent (such as acetonitrile, alcohol, ethanol, dimethyl sulfoxide, dimethylformamide and the like), such as a water/acetonitrile suspension, suction-filtering the obtained suspension, and drying the filter cake to obtain the crystal form V of the compound shown in the formula (1).

In the preparation methods of the crystal form II, the crystal form III, the crystal form IV and the crystal form V of the compound shown in the formula (1), the base is organic base or inorganic base, and the acid is inorganic acid or organic acid.

The organic base is selected from, for example, but not limited to, l-arginine, betaine, choline, diethylamine, lysine, n' -dibenzylethylenediamine, 2- (diethylamino) ethanol, 2-aminoethanol, 1- (2-hydroxyethyl) pyrrole, diethanolamine, di -ylethanolamine, n-methylglucamine, tromethamine, triethanolamine, 4- (2-hydroxyethyl) morpholine, imidazole, ethylenediamine, and the like.

The inorganic acid is selected from, for example, but not limited to, hydrobromic acid, hydrochloric acid, sulfuric acid, sulfurous acid, nitric acid, phosphoric acid, and the like. The organic acid is selected from, for example, but not limited to, sulfonic acid, dodecylsulfuric acid, 2-naphthalenesulfonic acid, benzenesulfonic acid, oxalic acid, 2-dichloroacetic acid, glycerophosphoric acid, 2-hydroxyethanesulfonic acid, L-aspartic acid, maleic acid, ethanesulfonic acid, 1, 5-disulfonic acid naphthalene, ethane-1, 2-disulfonic acid, cyclohexylsulfamic acid, p- benzenesulfonic acid, and the like.

The crystal form II, the crystal form III, the crystal form IV and the crystal form V of the compound shown in the formula (1) can be mutually transformed under certain conditions.

The crystal form III contains 4 water molecules, can be converted into the crystal form II after being completely dried, and does not contain crystal water.

During the transformation of form III to form II, different transition forms can be observed. After the drying process (natural air drying for about 8-15 minutes, suction filtration drying for about 1 minute), a transition state of crystal form X can be observed, wherein the crystal form X uses Cu-Ka radiation, and has characteristic peaks at 5.6 +/-0.2, 21.1 +/-0.2, 6.1 +/-0.211.0 +/-0.2, 10.1 phi 0.2, 23.0 +/-0.2, 18.5 +/-0.2, 9.0 phi 0.2, 26.0 +/-0.2 and 26.9 +/-0.2 in an X-ray powder diffraction pattern expressed by an angle of 2 theta; continued drying (about 1 hour) can be observed as the appearance of form V, and continued thorough drying (about 50 minutes) can yield a single form II.

The process of form II → form beta → form V-form II is a process of losing water of crystallization. In the process, the single crystal forms of the crystal form X and the crystal form V are difficult to exist stably for a long time, mixed crystals (including the crystal form X and the crystal form V) are easy to form in the conversion process, and the single crystal form X and the crystal form V can be converted into the single crystal form II repeatedly if the single crystal form X and the mixed crystals are placed in the natural air for a long time.

The inventors stirred form II in a mixed system of acetonitrile/water in order to prepare a single form V. The crystal form III can be obtained after long-time stirring (for example, more than 5 hours, for example, 9, 10, 11, 12 hours, etc.), but mixed crystal is obtained after shorter-time stirring (for example, less than 5 hours, for example, 2 hours), wherein the crystal form y comprises the crystal form III, the crystal form II, the crystal form V and a transition state, and the crystal form y is similar to the characteristic peak of the crystal form X but can not be determined to be the same crystal form; the mixed crystal can be prepared into a single crystal form III if the mixed crystal is continuously stirred (about 10 hours), can be prepared into a mixed crystal containing the crystal form V and the crystal form II if the crystal form III, the crystal form II, the crystal form V and the crystal form upsilon of the transition state are naturally air-dried or suction-filtered and dried, can be prepared into the mixed crystal mainly containing the crystal form V and the crystal form II by repeating the stirring in an acetonitrile/water mixed system for a short time and then naturally air-dried or suction-filtered and dried, and can be prepared into the single crystal form V by repeating the conversion process for multiple times mainly containing the crystal form V and the crystal form II.

Form V is stable in natural storage and does not significantly convert to form II when stored under vacuum drying at room temperature. The crystal form V is stirred in a mixed system of acetonitrile/water for more than 10 hours (for example, 12 hours and 16 hours), so that a single crystal form III can be obtained, and the single crystal form II can be prepared by fully drying the III by the method described above.

Form II, form Y → form III is a process of increasing water of crystallization;

form 11 → form γ → form V is a process of increasing and then losing water of crystallization.

The crystal form IV of the invention contains 1 water molecule and 1 methanol molecule, and can be converted into the crystal form II under the vacuum drying condition. The invention also provides a pharmaceutical composition which contains any one or more of amorphous powder, crystal form II, crystal form III, crystal form IV and crystal form V of the compound shown in the formula (1).

The pharmaceutical composition contains any one or more of amorphous powder, crystal form II, crystal form III, crystal form IV and crystal form V of the compound shown in the formula (1) and one or more pharmaceutical carriers and/or diluents. The composition can be prepared into any clinically or pharmaceutically acceptable dosage form, preferably oral preparations and injections.

The amorphous powder, form II, form III, form IV or form V of the compound of formula (1) of the present invention can be administered to a mammal, such as a human, by oral, parenteral (intravenous, intramuscular, subcutaneous or rectal), topical, etc. administration.

The dosage range of the amorphous powder, the crystal form II, the crystal form III, the crystal form IV or the crystal form V of the compound shown in the formula (1) or the mixture of two or more of the amorphous powder, the crystal form II, the crystal form III, the crystal form IV or the crystal form V is about 250-1000 mg/day, and the human body weight is about 4-20 mg/kg/day calculated according to 60 kg.

For parenteral administration, amorphous powders, crystalline forms of the compound of formula (1) of the present invention may be administered

II. The crystal form III, the crystal form IV or the crystal form V or a mixture of two or more of the crystal form III, the crystal form IV or the crystal form V is prepared into an injection, and comprises a sterile solution type, an emulsion type, a dispersion liquid type or a suspension liquid type preparation for intramuscular injection, intravenous drip, subcutaneous injection and the like, and sterile powder or concentrated solution for injection which is prepared or diluted into solution, dispersion liquid or suspension liquid before use.

The injection can be produced by conventional methods in the pharmaceutical field, and can be selected from aqueous solvents and non-aqueous solvents. The most commonly used aqueous solvent is water for injection, and 0.9% sodium chloride solution or other suitable aqueous solution can also be used; the nonaqueous solvent used in common use is vegetable oil, such as soybean oil for injection, and other aqueous solutions of ethanol, propylene glycol, polyethylene glycol, and the like. When the injection is prepared, appropriate additives such as osmotic pressure regulator, pH regulator, solubilizer, filler, antioxidant, bacteriostatic agent, emulsifier, suspending agent, etc. can be optionally added. Commonly used osmo-regulators include sodium chloride,' glucose, potassium chloride, magnesium chloride, calcium chloride, sorbitol, etc., preferably sodium chloride or glucose; commonly used pH regulators include acetic acid-sodium acetate, lactic acid, citric acid-sodium citrate, sodium bicarbonate-sodium carbonate, etc.; commonly used solubilizers include polysorbate 80, propylene glycol, lecithin, polyoxyethylene castor oil, and the like; commonly used bulking agents include lactose, mannitol, sorbitol, dextran, and the like; common antioxidants include sodium sulfite, sodium bisulfite, sodium metabisulfite, etc.; common bacteriostats are phenol, cresol, chlorobutanol and the like.

The pharmaceutical compositions may also be formulated for rectal or topical administration in conventional manner, including suppositories, ointments, creams, patches, powders, sprays, inhalants and the like.

For oral administration, the amorphous powder, form II, form III, form IV or form V of the compound of formula (1) of the present invention, or a mixture of two or more thereof may be formulated into conventional solid preparations such as tablets, capsules, pills, granules, etc. by conventional methods; it can also be made into oral liquid, such as oral solution, oral suspension, syrup, etc. The tablet is mainly oral common tablet, and comprises buccal tablet, sublingual tablet, buccal patch, chewable tablet, dispersible tablet, soluble tablet, effervescent tablet, release tablet, controlled release tablet, enteric coated tablet, etc. The capsules may be classified into hard capsules, soft capsules, release capsules, controlled-release capsules, enteric capsules and the like according to their dissolution and release characteristics. The pill includes dripping pill, sugared pill, pellet, etc. Granules can be divided into soluble granules, suspension granules, effervescent granules, enteric granules, release granules, controlled release granules and the like.

In the preparation of the oral preparation, suitable fillers, binders, disintegrants, lubricants, etc. may be added. Common fillers include starch, sugar powder, calcium phosphate, calcium sulfate dihydrate, dextrin, microcrystalline cellulose, lactose, pregelatinized starch, mannitol, and the like; common binders include sodium carboxy -based cellulose, PVP K30, hydroxypropyl cellulose, starch slurry, -based cellulose, ethyl cellulose, hydroxypropyl cellulose, gelatinized starch, and the like; common disintegrants include dry starch, crospovidone, croscarmellose sodium , sodium starch , low substituted hydroxypropylcellulose, etc.; common lubricants include magnesium stearate, talc, sodium lauryl sulfate, micronized silica gel, and the like.

On the other hand, the invention also provides application of amorphous powder, crystal form II, crystal form III, crystal form IV or crystal form V of the compound shown in the formula (1) or a mixture of two or more of the amorphous powder, the crystal form II, the crystal form III, the crystal form IV or the crystal form V in the aspect of preparing a medicament for treating and/or preventing infectious diseases.

In a further aspect, the present invention also provides a method for the treatment and/or prophylaxis of infectious diseases, comprising the step of administering an amorphous powder, form II, form III, form IV or form V of a compound of formula (1) according to the invention, or a mixture of two or more thereof, to a mammal, such as a human, in need of such treatment or prophylaxis.

The compound of formula (1) has excellent antibacterial activity on gram-positive bacteria, gram-negative bacteria, aerobic bacteria and anaerobic bacteria, has unexpected ultra-long half-life period, is stable to β -lactamase and DHP-1, can be safely used for treating and/or preventing various diseases caused by pathogenic microorganisms of various mammals (such as mice, rats, rabbits, dogs, cats, cattle, pigs and the like), including human beings, such as respiratory tract infection, urinary tract infection and the like

FIG. 1 is an X-ray powder diffraction pattern of crystalline form II of the compound of formula (1) with diffraction intensity (CPS) on the ordinate and diffraction angle (2 Θ) on the abscissa.

FIG. 2 is a DSC of form II of compound of formula (1) with power (mW) on the ordinate and temperature (. degree.C) on the abscissa.

: FIG. 3 is a single crystal X-ray unimolecular structure of form III of the compound of formula (1).

FIG. 4 is an X-ray powder diffractogram of form III of the compound of formula (1) with diffraction intensity (CPS) on the ordinate and diffraction angle (2 Θ) on the abscissa.

FIG. 5 is a single crystal X-ray unimolecular structure diagram of form IV of the compound of formula (1).

FIG. 6 is an X-ray powder diffraction pattern of form IV of the compound of formula (1) with diffraction intensity (CPS) on the ordinate and diffraction angle (2 Θ) on the abscissa.

FIG. Ί is an X-ray powder diffraction pattern of compound of formula (1) form V, with diffraction intensity (CPS) on the ordinate and diffraction angle (2 Θ) on the abscissa.

FIG. 8 is a DSC of form V of compound of formula (1) with power (mW) on the ordinate and temperature (. degree.C) on the abscissa.

FIG. 9 is an X-ray powder diffraction pattern of an amorphous powder of the compound of formula (1) with diffraction intensity (CPS) on the ordinate and diffraction angle (2. theta.) on the abscissa.

FIG. 10 is an X-ray powder diffraction pattern of crystalline form I of the compound of formula (1) with diffraction intensity (CPS) on the ordinate and diffraction angle (2 Θ) on the abscissa.

Figure 11 is a DSC plot of compound of formula (1) form I with power (mW) on the ordinate and temperature (° C) on the abscissa.

Detailed Description

The present invention will be described in further detail with reference to examples. These examples are intended to illustrate, but not to limit, the present invention. All the technologies that can be realized based on the above contents belong to the scope of the present invention.

^!HNMR was performed at 600 MHz using Bruker Avance 600 MHz (Bruker, Germany) with DMSO-d₆Measured as a solvent.

The X-ray powder diffraction (XRPD) pattern was measured on a D/max-RB 12kw rotating target X-ray diffractometer (Nippon Denshoku Co., Ltd.) using CuKa, 40 kV, 100mA,

2-Theta, 3 ℃ to 45V, 4 deg/min. And in the Differential Scanning Calorimetry (DSC), a DZ3335 differential scanning calorimeter (Nanjing Dazhan) is used for measuring under the conditions of a temperature rise rate of 10 ℃/min, a measuring temperature range of 25 ℃ -230 ℃, a nitrogen atmosphere and a flow rate of 40 ml/min.

Examples

Preparation example I: preparation of amorphous powder of compound of formula (1):

compound (1) was prepared according to the following synthetic route:

examples of preparation of intermediates

Preparation of AJ2S, 4R) -2-carboxylic acid-4-hydroxy-1- (p-nitrobenzyloxycarbonyl) pyrrolidine (intermediate b) in a reaction flask 880g (6.72 mol) L-hydroxyproline was dissolved in 6.72L of 2mol/L sodium hydroxide solution, a solution of 1305g (6.05 mol) p-nitrobenzyl chloro in 4000mL of dichloro alkane was added dropwise in ice bath, the temperature was controlled not to exceed 10 ℃, and after stirring for 3h the reaction, the aqueous phase was separated. Washing the water phase with dichloro alkane, acidifying with concentrated sulfuric acid until the pH is less than 7, precipitating a large amount of white solid, filtering, washing the filter cake with water, and vacuum drying to obtain 1523g of the product (2S,4R) -2-carboxylic acid-4-hydroxy-1- (p-nitrobenzyloxycarbonyl) pyrrolidine with the yield of 81.1%. B. Preparation of (2S,4R) -2- "Furan-2-yl -yl" amine acyl 1-4-hydroxy-1- (p-nitrobenzyloxycarbonyl) pyrrolidine (intermediate C)

In a dry reaction flask, 124g (0.4 mol) of (2S,4R) -2-carboxylic acid-4-hydroxy-1- (p-nitrobenzyloxycarbonyl) pyrrolidine were dissolved in 400 mL of dichloromethane. 58g (0.57 mol) triethylamine were added at-25 ℃ and the reaction was stirred to dissolve. slowly adding 69g (0.56 mol) of isopropyl chloride dropwise, and stirring at-25 ℃ for 1.5 h. A solution of 38.8g (0.4 mol) furan-2-yl amine in acetone (ca. 30 mL) was added below 0 ℃ and the reaction was continued lh. Then, lmol/L hydrochloric acid is added dropwise to acidify until the pH value is less than 7, ethyl acetate is used for extraction, and an organic phase is washed by water and a saturated sodium chloride solution in sequence and is concentrated under reduced pressure. To the residue was added 5mol/L hydrochloric acid for acidification, followed by stirring for 2 hours, then made basic with dilute alkali NaOH solution 0.1-2 mol/L to precipitate a solid, the reaction solution was suction filtered, and the cake was vacuum dried to obtain 135.1g of (2S,4R) -2- [ (furan-2-yl) amino acyl ] -4-hydroxy-1- (p-nitrobenzyloxycarbonyl) pyrrolidine, yield: 86.7 percent.

Preparation of C (2S,4R) -2- "Furan-2-ylmethyl" amine acyl ] -4- sulfonyl' oxy-1- (p-nitrobenzyloxycarbonyl) pyrrolidine (intermediate d)

To a dry reaction flask were added 75g (0.19 mol) of (2S,4R) -2- [ (furan-2-yl yl) carbamoyl ] -4-hydroxy-1- (p-nitrobenzyloxycarbonyl) pyrrolidine, 25g (0.25 mol) triethylamine and 200mL of dichloro alkane. A solution of 25.2g (0.22 mol) of methanesulfonyl chloride in dichloro alkane (20 mL) was slowly added dropwise at 0 ℃ and the reaction was continued for 1.5 h. After the reaction solution was washed with dilute hydrochloric acid (1 mol/L), water and saturated sodium bicarbonate solution in this order, the organic phase was evaporated to dryness under reduced pressure to obtain 79.5g of (2S,4R) -2- [ (furan-2-yl -yl) carbamoyl ] -4- sulfonyloxy-1- (p-nitrobenzyloxycarbonyl) pyrrolidine, the yield of which was 89.5%.

Preparation of K2S,4S) -2- [ (furan-2-yl yl) amine acyl ] -4-acetylthio-1- (p-nitrobenzyloxycarbonyl) pyrrolidine (intermediate e)

A mixed solution of 26.2g (56 mmol) of (2S,4R) -2- [ (furan-2-yl yl) amine acyl ] _4- sulfonyloxy-1- (p-nitrobenzyloxycarbonyl) pyrrolidine, 10.5g (92 mmol) of potassium thioacetate, 100 mL of N, N-bis yl amide (DMF) and 100 mL of benzene is added into a reaction flask, stirred, heated to 65-75 ℃ under a nitrogen atmosphere, and reacted for 6 hours. After the reaction, water was added to the reaction solution, the organic layer was separated, the aqueous layer was extracted with toluene, and the organic layers were combined. The organic layer was washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered with suction, and the filtrate was evaporated to dryness under reduced pressure to give 20.5g of (2S,4S) -2- [ (furan-2-yl -yl) amine acyl ] -4-acetylthio-1- (p-nitrobenzyloxycarbonyl) pyrrolidine as a red solid in a yield of 81.6%.

Preparation of K2S,4S) -2- [ (furan-2-yl yl) carbamoyl ] -4-mercapto-1- (p-nitrobenzyloxycarbonyl) pyrrolidine (intermediate f)

21.6g (48 mmol) of (2S,4R) -2- [ (furan-2-yl yl) amine acyl ] -4-acetylmercapto-1- (p-nitrobenzyloxycarbonyl) pyrrolidine was added to 125 mL of alcohol, stirred to dissolve, cooled to 0 to 5 ℃, and 13.5 mL of 4mol/L sodium hydroxide solution was added dropwise and reacted at 0 to 15 ℃ for 15 min. After the reaction, the reaction solution was adjusted to pH 10 with 4mol/L sodium hydroxide solution, extracted with dichloro alkane, the organic layer was discarded, the aqueous layer was adjusted to pH 1 by dropwise addition of 4mol/L dilute hydrochloric acid, extracted with ethyl acetate, the organic phase was washed with saturated sodium chloride solution, dried with anhydrous sodium sulfate and evaporated to dryness to obtain an oil, which was recrystallized from ethyl acetate-ether to give 17.1g of (2S,4S) -2- [ (furan-2-yl yl) carbamoyl ] -4-mercapto-1- (p-nitrobenzyloxycarbonyl) pyrrolidine as a powdery solid with a yield of 87.9%.

F. (4R,5S,6S) -34(3S, 5S) -N- (p-nitrobenzyloxycarbonyl) -5- "(furan-2-yl -yl) amine acyl ] -3-pyrrolidine ] thio-6- [ (R) -l-hydroxyethyl ] -4- -yl-7-oxo-1-azabicyclo

Preparation of p-nitrobenzyl [3.2.0] hept-2-ene-2-carboxylate (intermediate g)

73g (180 mmol) (2S,4S) -2- [ (furan-2-ylmethyl) carbamoyl ] -4-mercapto-1- (p-nitrobenzyloxycarbonyl) -pyrrolidine and 120g (200 mmol) p-nitrobenzyl [ (4R,5S,6S) -3- [ (diphenylphosphinoyl) oxy ] -6- [ (R) -l-hydroxyethyl) -4- yl-7-oxo-azabicyclo [3.2.0] hept-2-ene-2-carboxylate (MAP) are dissolved in 350 mL N, N-di yl amide and the temperature is reduced to-15 ℃ and 18.7g triethylamine are added dropwise in a nitrogen atmosphere; reacting at-15 ℃ for 2h, and finishing the liquid-phase color phrase detection reaction. 1600 mL of ethyl acetate was added to the reaction solution; the organic layer was separated, washed successively with 800 mL of water, 200mL of 5% hydrochloric acid and 200mL of saturated sodium bicarbonate, dried over anhydrous sodium sulfate, concentrated, crystallized, filtered and the filter cake dried under vacuum to give 74.6g of the title compound in 55.3% yield. Preparation of Compound of formula (1)

Preparation of (4R,5S,6S) -3- [ (3S,5S) -5- [ (furan-2-yl yl) amine acyl ] -3-pyrrolidine ] thio-6- [ (R) -l-hydroxyethyl ] -4- yl-7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylic acid (compound of formula (1))

40g (53 mmol) of (4R,5S,6S) -3- [ (3S,5S) -N- (p-nitrobenzyloxycarbonyl) -5- [ (furan-2-yl) -ylidene amino -acyl radical]-3-pyrrolidines]Thio-6- [ (R) -l-hydroxyethyl]-4-methyl-7-oxo-1-azabicyclo [3.2.0]Dissolving p-nitrobenzyl hept-2-ene-2-carboxylate in 160 mL tetrahydrofuran; another 120 mL of water was added with 11.2g of sodium bicarbonate. The two solutions were mixed, 4.0g of anhydrous ordinary palladium on charcoal was added to the mixture, and hydrogenated at high pressure l-6MPa for 2 hours. Filtering the reaction solution to remove palladium-carbon; the filtrate was washed three times with 200mL of ethyl acetate, the aqueous layer was separated and brought to dryness with acetic acid under cooling in an ice bath_pThe H value was 5.5 and it was purified by passing it through the preparative liquid phase (CHP-20P macroporous resin) and the collected fractions containing the product were lyophilized to give 6.6g of the compound of formula (1) as an amorphous powder with a yield of 28.4%. Using it as a starting material for the preparation of polymorphic forms of the compound of formula (1)。

The molecular formula is as follows: c₂。H₂₅N₃0₆S

Molecular weight: 435.49

!HNMR (DMSO-d₆, 600MHz) δ: 1.12(d, 3Η), 1.15(d, 3H), 1.55(dt, IH), 2.55(dt, IH), 2.65(dd, IH), 3.17(t, IH), 3.37(m, 2H), 3.57(m, IH), 3.80(t, IH), 3.94(m, I H), 4.12(d, IH), 4.29(m, 2H), 6.23(d, IH), 6.38(s, IH), 7.56(s， 1H)， 8.50(t, 1H)。

XRPD diffraction: the XRPD diffraction measurements are shown in fig. 9. Preparation of polymorph of Compound of formula (1)

Comparative example 1 preparation of Compound form I of formula II)

1.02g of the compound of formula (1) prepared in preparation I above was weighed and stirred until dissolved in 50mL of methanol, and then stirred until crystals precipitated. Filtration and drying of the filter cake in vacuo for 12 hours gave 0.34g of crystals. This crystal was designated as form I crystal, i.e. compound form I of formula (1), which was identical to the compound of formula (1), i.e. compound a, prepared as described in example 1 of WO 2009/000210 a 1.

XRD, the XRD measurement result is shown in figure 10;

DSC maximum decomposition temperature: 164.6 ℃ and the results are shown in FIG. 11. Example 1 preparation of crystalline form II of the Compound of formula II)

0.20 g of the amorphous powder of the compound of formula (1) prepared in preparation example I above was weighed, 1 mL of water was added thereto and stirred to suspend, then 0.047 g of solid sodium bicarbonate was added thereto and stirred to form a solution, then 2mol/L hydrochloric acid was added dropwise to adjust the resulting solution to pH =6.0, and crystals were precipitated and stirred for about 0.5 hour. Filtration and vacuum drying of the filter cake for 12 h gave 0.065 g of crystals. The XRPD diffraction measurements are shown in figure 1; DSC maximum decomposition temperature: 169.5 ℃, the results are shown in fig. 2. This crystal is designated as form II crystal, i.e. compound of formula (1) form II. Example 2 preparation of crystalline form II of the Compound of formula II)

0.30 g of the amorphous powder of the compound of formula (1) prepared in preparation example I above was weighed, 2mL of water was added thereto and stirred to suspend, then 0.07 g of sodium bicarbonate solid was added and stirred until a solution was formed, then alcohol of 45% by volume was added, then 2N hydrochloric acid was added dropwise to the resulting mixture to adjust pH =6.0, crystals were precipitated, and stirring was continued for about 0.5 hour. Filtration and drying of the filter cake in vacuo for 12 hours gave 0.105 g of crystals. The XRPD diffractogram thereof corresponds to the diffractogram of form II of compound of formula (1) as shown in fig. 1, with the diffraction angles (2 Θ) having peaks at the following positions: 6.60, 10.96, 12.36, 13.32, 14.02, 15.18, 15.70, 16.94, 19.74, 20.80, 21.26, 21.64, 22.12, 23.04, 24.42, 25.12, 26,60, 26.94, 28.42, 28.80. EXAMPLE 3 preparation of Compound of formula (1) in crystalline form II

0.4 g of the amorphous powder of the compound of formula (1) prepared in preparation example I above was weighed, lmL water was added thereto and stirred to suspend, and then 2mL of THF was added thereto and stirred until it was dissolved. Then, THF was distilled off under reduced pressure, the resulting mixture was filtered, and the filtrate was left to stand at 0 ℃ for 12 hours in a refrigerator. The precipitated crystals were filtered off and dried in vacuo for 12 hours to give 0,07 g of crystals. The XRPD diffractogram thereof corresponds to the diffractogram of form II of compound of formula (1) as shown in fig. 1, with the diffraction angles (2 Θ) having peaks at the following positions: 6.46, 10.92, 13.16, 13.98, 15.08, 15.58, 16.78, 19.50, 20.76, 21.18, 21.56, 22.04, 22.94, 24.48, 24.94, 25.68, 26.52, 26.66, 27.42, 28.76. Example 4 preparation of crystalline form Π of compound (upsilon)

1.00 g of the amorphous powder of the compound of formula (1) prepared in preparation example I above was weighed, 4mL of water and 4mL of THF were added thereto, and stirred until it was dissolved. Then, 14mL of isopropyl alcohol was added dropwise to precipitate crystals. After stirring vigorously for 1 hour, it was filtered. The filter cake was dried under vacuum for 12 hours to give 0.28 g of crystals. The XRPD diffraction pattern of the compound conforms to the diffraction pattern of the compound of the formula (1) in the crystal form II as shown in figure 1, and the diffraction angles (2 theta) have peaks at the following positions of 6.54, 11.00, 13.24, 14.08, 15.16, 15.74, 16.88, 19.50, 20.84, 21.28, 22.10, 23.04, 23.80, 24.52, 25.02, 26.20, 26.52, 28.88 and 30.44. Example 5 preparation of crystalline form II of the Compound of formula II)

0.20 g of an amorphous powder of the compound of formula (1) prepared in preparation example I above was weighed, 2mL of water was added thereto and stirred to suspend, and then heated to 50-60 ℃ until it was dissolved, and the resulting solution was transferred to an ice water bath to be cooled for 1 hour to precipitate crystals. The precipitated crystals were filtered off and dried in vacuo for 12 hours to give 0.055 g of crystals. The XRPD diffractogram thereof corresponds to the diffractogram of form II of compound of formula (1) as shown in fig. 1, with the diffraction angles (2 Θ) having peaks at the following positions: 6.54, 10.98, 12.34, 13.26, 14.06, 15.18, 15.72, 16.92, 19.68, 20.80, 21.26, 21.62, 22.12, 23.04, 23.78, 24.60, 25.00, 25.68, 26.52, 26.68, 28.92. Example 6 preparation of crystalline form III of compound of formula Π Μ an ancient type of spoon 0.6 g of the amorphous powder of compound of formula (1) prepared above in preparation I is weighed into it 1.2 mL of water and 0.9 mL of acetonitrile and heated until the compound dissolves. The obtained solution was filtered, and the filtrate was placed in a 0 ℃ refrigerator and left to crystallize. After 24 hours crystals of a size suitable for single crystal testing were obtained. The XRPD diffraction measurements are shown in figure 4. This crystal is designated as form III crystal, i.e. form III of the compound of formula (1).

The unit cell parameters of the compound of formula (1) in crystal form III are as follows:

the single crystal X-ray unimolecular structure of form III of the compound of formula (1) is shown in figure 3.

Example 7 preparation of form IV of the Compound of formula II)

0.60 g of an amorphous powder of the compound of formula (1) prepared in preparation example I above was weighed, 3 mL of water was added thereto and stirred to suspend, then 0.14 g of sodium bicarbonate solid was added and stirred until a solution was formed, 2mL of alcohol was added, then 2mol/L of HC1 was added dropwise to adjust the mixture to pH =6.0, the resulting solution was membrane-filtered, the filtrate was left to crystallize in a 0 ℃ refrigerator, and crystals having a size suitable for single crystal diffraction appeared after 24 hours. The XRPD diffraction measurements are shown in fig. 6. This crystal is designated as form IV crystal, i.e. form IV of the compound of formula (1).

The unit cell parameters of compound form IV of formula (1) are as follows:

value of a parameter

Crystal system orthogonality

Space group p iota a (A) 8.48

b (A) 10. 00

c (A) 27. 16

Cell parameters

a (。 ) 90

β (。 ) 90

Y (。 ) 90

Unit cell volume v (A)³) 2307. 64

z (number of molecules in unit cell) 4

Calculated Density (g/cm)³) 1.621A single crystal X-ray unimolecular structure of form IV of the compound of formula (1) is shown in FIG. 5.

Example 8 formula II) preparation of Compound form II crystalline form II

Weighing 1g of the compound of the formula (1) in the crystal form II (crystals prepared in examples 1 to 5), adding 3 mL of acetonitrile and 4mL of water, stirring to form a suspension, stirring the suspension for 16 hours, and performing suction filtration to obtain the crystal form III. Example 9 formula II) Compound Crystal form II preparation of Crystal form V

L g the crystal form II of the compound of formula (1) prepared in the above example 1 is weighed, 3 mL acetonitrile and 4mL water are added into the crystal form II, the mixture is stirred to suspend, the suspension is stirred for 2 hours, filtered and dried to obtain a mixed crystal of the crystal form II and the crystal form V, the mixed crystal is repeatedly stirred in an acetonitrile/water mixed system for 2 hours, filtered and dried to obtain a mixed crystal mainly comprising the crystal form II and the crystal form V, the mixed crystal is repeatedly (more than 2 times) subjected to the conversion process, and the vacuum drying is carried out to obtain 0.2g of crystal. The XRPD diffraction measurements are shown in fig. 7; DSC maximum decomposition temperature: 148.7 ℃, and the results are shown in fig. 8. This crystal is designated as form V crystal, i.e. form V of the compound of formula (1). EXAMPLE 10 formula II) Compound form V preparation of form III

Suspending and stirring the crystal form V (the crystal prepared in example 9) in a mixed system of acetonitrile/water (3: 4) for 12 hours, and filtering the suspension to obtain the crystal form III, wherein the crystal form in of the compound shown in the formula (η) in example 11 is converted into the crystal form II

And carrying out suction filtration on the crystal form III (crystal precipitated after the crystal is placed for 24 hours in the example 6), and carrying out vacuum drying for 90 min to obtain a crystal form II. Example 12 conversion of form IV of the Compound of formula II) to form II

The compound of formula (1) in crystal form IV (crystal obtained in example 7) was placed in a vacuum drying oven and vacuum dried overnight to obtain crystal form II.

Determination of the in vitro antibacterial Activity of amorphous powders and polymorphs of the Compound of formula (1)

The test strain is a standard strain which is purchased from the market; clinically isolating the strain: is commercially available.

The experimental method comprises the following steps: adopts standard agar double dilution method, and refers to pharmacological experiment methodology

P1659-1660, national public health Press, eds: xu Tertiary cloud, etc., 1 month, 3 rd edition 2002.

The experimental results are shown in tables 1 and 2 below. In vitro antibacterial Activity of amorphous powder of Compound of formula (1)

Clinical isolation of amorphous powder MIC of Strain Compound of formula (1)₉Q (mg/L) Klebsiella pneumoniae 0.062

Pseudomonas aeruginosa 16

Staphylococcus aureus 0.125

The result of a 0.25 staphylococcus epidermidis experiment shows that the amorphous powder of the compound shown in the formula (1) has better antibacterial activity on clinical isolate bacteria. TABLE 2 in vitro antibacterial Activity of Compound of formula (1) form II

Standard Strain Compound of formula (1) form II MIC (mg/L) Escherichia coli ATCC 259220.016

Klebsiella pneumoniae ATCC 7006030.031

Enterobacter cloacae P990.016 Pseudomonas aeruginosa ATCC 278538

Haemophilus influenzae ATCC 492470.5

Staphylococcus aureus ATCC 292130.062

Staphylococcus epidermidis, ATCC 122280.031

Streptococcus pyogenes ATCC 196150.004

Enterococcus faecalis ATCC 292121

Bacteroides fragilis ATCC 252850.125

Clinical isolation of Strain formula (1) Compound Crystal form II MIC₉₀(mg/L) Staphylococcus aureus 0.125

Staphylococcus epidermidis 0.25

Streptococcus pneumoniae 0.016

Proteus mirabilis 0.031

Moraxella catarrhalis 0.004

Haemophilus influenzae 0, 125

Enterobacter cloacae 0.25

The results of the Klebsiella pneumoniae 0.062 experiment show that the crystal form II of the compound shown in the formula (1) has better antibacterial activity on standard gram-positive bacteria, negative bacteria and anaerobic bacteria and clinically isolated gram-positive bacteria, negative bacteria and anaerobic bacteria. SD rat in vivo pharmacokinetic test of crystal form II of compound of formula (1)

Sample of the inventive Compound of formula (1) Crystal form II (self-made)

The experimental animal is male SD rat with weight of 200-250 g, and purchased from market.

Experimental method according to the method for measuring the Effect of WO 2009/000210A 1 in "pharmacokinetic data in SD rat

Experimental results half-life period t_1/2(h) It was 0.36. + -. 0.01 (X earth SD).

The experimental result shows that the crystal form II of the compound shown in the formula (1) has good plasma half-life and good pharmacokinetic property. Physical Properties of polymorphic forms of the Compound of formula (1)

Experimental example 1 method for examining residual solvents of the compound of formula (1) in crystal form II and crystal form I according to the present invention, the test was performed according to the residual solvent assay method (annex beautiful P of the second part of the chinese pharmacopoeia 2005 edition). A capillary column using (5%) phenyl- (95%) -based polysiloxane as stationary liquid (or stationary liquid with similar polarity) was used as a chromatographic column, and the detector was a hydrogen Flame Ionization Detector (FID). Calculated as peak area by external standard method.

The results are shown in Table 3 below.

Table 3 residual crystalline form II of the compound of formula (1) of the present invention is aligned with that of crystalline form I:

and (4) conclusion: the content of methanol in the crystal form I obviously exceeds the limit of quality standard, and the safety of the crystal form I in clinical application is hidden, while the content of the alcohol in the crystal form II is very low and is far lower than the limit specified by the quality standard. Experimental example 2 comparative investigation conditions of stability tests of the compound of formula (1) of the present invention in crystal form II and crystal form I include high temperature of 60 ℃, illumination of 4500Lx + -500 Lx,

the method comprises the steps of respectively carrying out stability tests on the crystal form II and the crystal form I of the compound shown in the formula (1) under the illumination experiment conditions of high temperature of 60 ℃ and 4500Lx +/-500 Lx, measuring the content of related substances (the total content of ring-opening substances and other impurities) and the content of main drugs under the experiment conditions, and taking the increase amplitude of the related substances and the decrease amplitude of the content of the compound shown in the formula (1) as investigation items.

Measuring related substances by high performance liquid chromatography according to appendix V D of second part of Chinese pharmacopoeia 2005 edition, and using octadecylsilane chemically bonded silica as filler; mobile phases of 0.03mol/L ammonium dihydrogen phosphate solution (pH adjusted to 4.0 with phosphoric acid) -acetonitrile (90: 10), and 0.03mol/L ammonium dihydrogen phosphate solution (pH adjusted to 4.0 with phosphoric acid) -acetonitrile (20: 80); the flow rate is 1.0ml per minute, and gradient elution is carried out; the detection wavelength is 220nm, and the peak area is calculated according to an external standard method. The content of the compound of the formula (1) is determined by high performance liquid chromatography according to appendix V D of the second part of Chinese pharmacopoeia 2005 edition. Octadecylsilane chemically bonded silica is used as a filling agent; 0.03mol/L ammonium dihydrogen phosphate solution (pH adjusted to 3.8 with phosphoric acid) -acetonitrile (88: 12) was used as a mobile phase, and the detection wavelength was 220 nm. The calculation formula is that the related substance increase amplitude = (the content of the related substance in 5 days or 10 days-the content of the related substance in 0 day)/the content of the related substance in 0 day X100 percent

The results of the reduction of the content of the compound of formula (1) = (content for 0 day-5 days or 10 days)/content for 0 day X100% are shown in tables 4 and 5 below. TABLE 4-1 high temperature investigation results of form I of Compound of formula (1)

TABLE 5-1 results of light examination of form I of Compound of formula (1)

And (4) conclusion:

as can be seen from the above tables 4-1 and 4-2, the related substance increase amplitude of the crystal form II is lower than that of the crystal form I at a high temperature of 60 ℃, the content reduction amplitude of the compound of the formula (1) is smaller than that of the crystal form I, and the obvious difference is obtained, which indicates that the stability of the crystal form II is better than that of the crystal form I at the high temperature of 60 ℃.

As can be seen from the above tables 5-1 and 5-2, the content of the compound shown in the formula (1) is reduced by a lower margin than that of the crystal form I when the crystal form II is illuminated at 4500Lx +/-500 Lx, and the difference is obvious, so that the stability of the crystal form II is better than that of the crystal form I when the crystal form II is illuminated at 4500Lx +/-500 Lx. Formulation examples of polymorphic forms of the Compound of formula (1)

Formulation example 1 preparation of sterile powder injection formulation of crystalline form II of Compound of formula (1) according to the invention

Mixing sterile crystal form II powder of compound of formula (1) with sterile anhydrous sodium carbonate powder, packaging, and capping. The present invention has been described in full detail in this specification. All the technologies that can be realized based on the above contents belong to the scope of the present invention. Furthermore, after reading the above-mentioned contents, those skilled in the art can make various modifications, changes or adaptations to the present invention within the scope not departing from the spirit of the present invention, and these modifications are also within the scope defined by the appended claims.

Claims

Claims to follow

1. A carbapenem derivative (4R,5S,6S) -3- [ (3S,5S) -5- [ (furan-2-yl yl) amine acyl ] -3-pyrrolidine ] thio-6- [ (R) -l-hydroxyethyl ] -4- yl-7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylic acid represented by formula (1) in the form of an amorphous powder:

2. form II crystals of (4R,5S,6S) -3- [ (3S,5S) -5- [ (furan-2-yl yl) amine acyl ] -3-pyrrolidine ] thio-6- [ (R) -l-hydroxyethyl ] -4- yl-7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylic acid, form II, characterized by an X-ray powder diffraction pattern using Cu-Ka radiation at 2 Θ angles having characteristic peaks at 6.5. + -. 0.2, 11.0. + -. 0.2, 16.8. + -. 0.2, 23.0S 0.2.
3. The crystalline form Π of claim 2 wherein the X-ray powder diffraction pattern using Cu-Ka radiation, expressed in degrees 2 Θ, further has characteristic peaks at 15.1 z 0.2, 15.7 ± 0.2.
4. The crystalline form II according to claim 3, further characterized by an X-ray powder diffraction pattern using Cu-Ka radiation expressed as an angle of 2 Θ at 13.3 ± 0.2, 21.1 ± 0.2, 22.1 te 0.2,

and the characteristic peak is at 0.2 of 24.6.
5. The crystalline form II according to any one of claims 2 to 4, characterized by an X-ray powder diffraction pattern substantially as shown in figure 1.
6. The crystalline form II according to any of claims 2 to 5, characterized by a DSC curve with a maximum decomposition temperature of 165-185 ℃ or as shown in FIG. 2.

7. (4R,5S,6S) -3- [ (3S,5S) -5- [ (furan-2-yl yl) amine acyl ] -3-pyrrolidine ] thio-6- [ (R) -l-hydroxyethyl ] -4- yl-7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylic acid

.4H₂0, form III crystal, form III, characterized by an X-ray powder diffraction pattern using Cu-Ka radiation at an angle of 2 Θ having characteristic peaks at 5.3 ± 0.2, 10.7 Θ 0.2, 15.7 ± 0.2, 21.3 ± 0.2.
8. The crystalline form III of claim 7, further characterized by an X-ray powder diffraction pattern using Cu-Ka radiation expressed at an angle of 2 Θ having characteristic peaks at 23.1 deg. 0.2,. 27.3 ± 0.2.
9. The crystalline form III of claim 8, further characterized by an X-ray powder diffraction pattern using Cu-Ka radiation expressed as an angle of 2 Θ at 20.7 ± 0.2, 23.5 Θ, 0.2, 27.0 ± 0.2, 2,

The characteristic peak is at 28.8 plus or minus 0.2.
10. The crystalline form III of any of claims 7-9, characterized by an X-ray powder diffraction pattern substantially as shown in figure 4.

11. (4R,5S,6S) -3- [ (3S,5S) -5- [ (furan-2-yl yl) amine acyl ] -3-pyrrolidine ] thio-6- [ (R) -l-hydroxyethyl ] -4- yl-7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylic acid

· Η₂0 · Η₃Form IV crystal of oh, form IV, characterized by an X-ray powder diffraction pattern using Cu-Ka radiation with characteristic peaks at 6.5, 10.9, 12.9, 0.2, 26.1 ± 0.2, expressed as angles of 2 Θ.
12. Form IV according to claim 11, characterized in that the X-ray powder diffraction pattern expressed in degrees 2 Θ using Cu-Ka radiation also has characteristic peaks at 32.8 ± 0.2, 39.6X 0.2.
13. Form IV according to claim 12, characterized in that the X-ray powder diffraction pattern expressed in degrees 2 Θ using Cu-Ka radiation also has characteristic peaks at 21.2 ± 0.2, 22.0 ± 0.2, 22.8 bar 0.2, 24.4 bar 0.2.
14. Form IV according to any one of claims 11 to 13, characterized by an X-ray powder diffraction pattern substantially as shown in figure 6.

15. Form V of (4R,5S,6S) -3- [ (3S,5S) -5- [ (furan-2-ylmethyl) carbamoyl ] -3-pyrrolidine ] thio-6- [ (R) -l-hydroxyethyl ] -4- yl-7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylic acid, form V, characterized by the characteristic peaks at 6.1S 0.2, 10.3S 0.2, 11.6. + -. 0.2, 20.8S 0.2 in an X-ray powder diffraction pattern expressed in degrees 2 Θ using Cu-Ka radiation.
16. Form V according to claim 15, characterized in that the X-ray powder diffraction pattern expressed in degrees 2 Θ using Cu-Ka radiation is also characterized at 16.5 ± 0.2, 21.5 Shih 0.2
17. Form V according to claim 16, wherein the X-ray powder diffraction pattern expressed in degrees 2 Θ using Cu-Ka radiation further has characteristic peaks at 9.0 sec 0.2, 19.5 sec 0.2, 19.8 ± 0.2, 22.2 sec 0.2.
18. Form V according to any one of claims 15 to 17, characterized by an X-ray powder diffraction pattern substantially as shown in figure 7.
19. Form V according to any one of claims 15 to 18, characterized by a DSC curve as shown in figure 8.

20. A crystalline mixture of (4R,5S,6S) -3- [ (3S,5S) -5- [ (furan-2-yl yl) carbamoyl ] -3-pyrrolidine ] thio-6- [ (R) -l-hydroxyethyl ] -4- yl-7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylic acid comprising the crystalline form II of any one of claims 2-6 and the crystalline form V of any one of claims 15-19.
21. A process for the preparation of a compound of formula (1) in the form of an amorphous powder according to claim 1, comprising:

(1 )

wherein PG_rX represents an amino protecting agent, PG₂X represents a carboxyl protecting agent, AA-X represents a hydroxyl activating agent, wherein PG! represents an amino protecting group, PG₂Represents a carboxyl protecting group, AA represents a leaving group, X represents a halogen selected from fluorine, chlorine, bromine and iodine; CH (CH)₃CO-S-M represents an alkali metal thioacetate, wherein M represents an alkali metal cation; mixing L-hydroxyproline with amino protective reagent PG_rX is subjected to amino protection reaction to form an intermediate b, then reacts with ^ in the presence of a coupling activator to form an intermediate C, reacts with a hydroxyl activator AA-X to form an intermediate d, reacts with alkali metal thioacetate to form an intermediate e, is subjected to deprotection reaction by alkali hydrolysis, and removes CH₃The CO-moiety forms an intermediate f with f under basic conditionsThe reaction forms intermediate g, which is then subjected to a deprotection reaction to yield the compound of formula (1).
22. A process for the preparation of crystalline form II according to any one of claims 2 to 6, which comprises adding a base to an aqueous suspension of an amorphous powder of (4R,5S,6S) -3- [ (3S,5S) -5- [ (furan-2-yl -yl) amine acyl ] -3-pyrrolidine ] thio-6- [ (R) -l-hydroxyethyl ] -4- yl-7-oxo-1-azabicyclo [3.2.0] hepta-2- -2-carboxylic acid until completely dissolved, optionally with or without the addition of a good solvent, adjusting the pH to below 7 with an acid, precipitating crystals, filtering and drying.
23. A process for the preparation of crystalline form II according to any one of claims 2 to 6, which comprises adding a good solvent to an aqueous suspension of an amorphous powder of (4R,5S,6S) -3- [ (3S,5S) -5- [ (furan-2-yl yl) amine acyl ] -3-pyrrolidine ] thio-6- [ (R) -1-hydroxyethyl ] -4-methyl-7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylic acid until the compound is completely dissolved, distilling off the good solvent under reduced pressure, filtering, precipitating crystals and drying.
24. A process for the preparation of crystalline form II according to any one of claims 2 to 6, which comprises adding a good solvent to an aqueous suspension of an amorphous powder of (4R,5S,6S) -3- [ (3S,5S) -5- [ (furan-2-yl yl) amine acyl ] -3-pyrrolidine ] thio-6- [ (R) -l-hydroxyethyl ] -4-methyl-7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylic acid until complete dissolution, adding a poor solvent, precipitating crystals, filtering and drying.

^:25. A process for the preparation of the crystalline form II as claimed in any of claims 2 to 6, which comprises reacting

(4R,5S,6S) -3- [ (3S,5S) -5- [ (furan-2-yl yl) carbamoyl ] -3-pyrrolidine ] thio-6- [ (R) -l-hydroxyethyl ] -4- yl-7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylic acid amorphous powder in water suspension is heated to complete dissolution, cooled, crystals precipitated, filtered and dried.
26. A process for the preparation of crystalline form III according to any one of claims 7 to 10, which comprises heating a water/acetonitrile suspension of an amorphous powder of (4R,5S,6S) -3- [ (3S,5S) -5- [ (furan-2-yl yl) amine acyl ] -3-pyrrolidine ] thio-6- [ (R) -l-hydroxyethyl ] -4-methyl-7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylic acid to dissolution, filtering, freeze-crystallization, filtering.
27. A process for the preparation of crystalline form IV, as claimed in any one of claims 11 to 14, which comprises adding a base to an aqueous suspension of an amorphous powder of (4R,5S,6S) -3- [ (3S,5S) -5- [ (furan-2-yl -yl) amine acyl ] -3-pyrrolidine ] thio-6- [ (R) -l-hydroxyethyl ] -4- yl-7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylic acid until completely dissolved, adding alcohol, adjusting the pH with an acid to 5 to 6, and cryocrystallizing.
28. A process for the preparation of crystalline form V according to any one of claims 15 to 19, characterized in that a suspension of an amorphous powder of (4R,5S,6S) -3- [ (3S,5S) -5- [ (furan-2-yl yl) amine acyl ] -3-pyrrolidine ] thio-6- [ (R) -l-hydroxyethyl ] -4-methyl-7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylic acid in water/acetonitrile is stirred, the resulting suspension is suction filtered and the biscuit is dried.
29. The method of claim 22 or 27, wherein the base is an organic base or an inorganic base, the acid is an inorganic acid or an organic acid, wherein the inorganic base is selected from potassium hydroxide, sodium hydroxide, zinc hydroxide, calcium hydroxide, potassium carbonate, potassium bicarbonate, sodium carbonate or sodium bicarbonate, and the organic base is selected from L-arginine, betaine, choline, diethylamine, lysine, and sodium bicarbonate,

N, N' -dibenzylethylenediamine, 2- (diethylamino) ethanol,. 2-aminoethanol, 1- (2-hydroxyethyl) pyrrole, diethanolamine, dimethylethanolamine, N- -glucamine, tromethamine, triethanolamine, 4- (2-hydroxyethyl) morpholine, imidazole or ethylenediamine, an inorganic acid selected from hydrobromic acid, hydrochloric acid, sulfuric acid, sulfurous acid, nitric acid or phosphoric acid, an organic acid selected from sulfonic acid, dodecylsulfuric acid, 2-naphthalenesulfonic acid, benzenesulfonic acid, oxalic acid, 2-dichloroacetic acid, glycerophosphoric acid, 2-hydroxyethanesulfonic acid, L-aspartic acid, maleic acid, ethanesulfonic acid, 1, 5-disulfonic naphthalene, ethane-1, 2-disulfonic acid, cyclohexylsulfamic acid or p-toluenesulfonic acid.
30. The method according to claim 22, 23 or 24, wherein the good solvent is selected from 5-90 vol% alcohol-water solution or tetrahydrofuran.
31. The method of claim 24 wherein the poor solvent is isopropanol.
32. The method of claim 25 wherein said heating is by heating to 50-60 ℃ an aqueous suspension of an amorphous powder of (4R,5S,6S) -3- [ (3S,5S) -5- [ (furan-2-yl yl) carbamoyl ] -3-pyrrolidine ] thio-6- [ (R) -l-hydroxyethyl ] -4- yl-7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylic acid and said reducing said temperature to less than 5 ℃.
33. A pharmaceutical composition comprising an amorphous powder of a compound of formula (1) as defined in claim 1 and any one or more of a crystal of a compound of formula (1) as defined in any one of claims 2 to 19 and a pharmaceutically acceptable carrier or diluent.
34. Use of the amorphous powder according to claim 1 and the crystal according to any one of claims 2 to 20 for the preparation of a medicament for the treatment and/or prevention of infectious diseases.