Beta-lactam compound as well as crystal form, preparation method and application thereof

Abstract

The present invention relates to a beta-lactam compound and a preparation method and use thereof, the beta-lactam compound has a structure represented by formula (I), n = 0-6, and the X-ray powder diffraction pattern 2theta diffraction angle of the crystal form A has characteristic diffraction peaks at 5.8 + / -0.2 degrees, 10.8 + / -0.2 degrees, 15.3 + / -0.2 degrees, 17.4 + / -0.2 degrees and 18.6 + / -0.2 degrees. The beta-lactam compound of the present invention not only has improved efficacy, but also has improved solubility and stability. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical chemistry technology, and in particular to a β-lactam compound, its crystal form, preparation method and use thereof. The present invention also relates to a composition comprising the β-lactam compound. Background Art

[0002] So far, a variety of antibacterial agents have been developed, such as β-lactams, aminoglycosides, tetracyclines, fluoroquinolones, glycopeptides, macrolides, etc. Patent application CN 113754651A discloses a novel β-lactam compound, which has good antibacterial properties and low drug resistance against bacteria, especially Gram-negative bacteria, and has good prospects for effectively treating various diseases. However, there is still a need to further improve its physical properties and efficacy. Summary of the invention

[0003] The present invention is made to overcome the above-mentioned deficiencies in the prior art.

[0004] The present invention provides a new β-lactam compound, which is a compound of formula (I):

[0005]

[0006] wherein n=0-6, preferably 2-3.

[0007] The compound of formula (I) of the present invention is in the form of a sodium salt. Surprisingly, it was found that the compound of the present invention has improved efficacy against Klebsiella pneumoniae compared to the free acid of the prior art. Specifically, the compound of the present invention exhibits a lower minimum inhibitory concentration.

[0008] In addition, the present invention also provides a new crystalline form A of the compound of formula (I), wherein the X-ray powder diffraction pattern of the crystalline form A is Figure 2 The θ diffraction angle has characteristic diffraction peaks at 5.8±0.2°, 10.8±0.2°, 15.3±0.2°, 17.4±0.2° and 18.6±0.2°.

[0009] Unexpectedly, it has been found that the crystalline form A can further improve the solubility and stability compared with the amorphous form of the compound of formula (I), thereby facilitating the preparation of a powder injection and the like for injection, etc. Therefore, when preparing an injection, etc., the present invention allows the use of less solvent to dissolve the sample, thereby helping to reduce costs and improve product utilization.

[0010] In addition, the present invention also provides a method for preparing the compound of formula (I), comprising:

[0011] The compound of formula (A-1) and the sodium conversion agent are dissolved in a good solvent to form a salt,

[0012]

[0013] In addition, the present invention also provides a method for preparing the crystalline form A of the compound of formula (I), which further comprises:

[0014] A poor solvent for the compound of formula (I) is added to the salt-formed solution at a temperature of 0°C-30°C to form crystals.

[0015] In addition, the present invention also provides a pharmaceutical composition, characterized in that it comprises the compound of formula (I) of the present invention or the compound of formula (I) prepared according to the method of the present invention, and a pharmaceutically acceptable auxiliary agent.

[0016] In addition, the present invention also provides the use of the compound of formula (I) of the present invention or the pharmaceutical composition of the present invention against Gram-negative bacteria, in particular Escherichia coli, Klebsiella pneumoniae, Klebsiella aerogenes, Enterobacter cloacae, Citrobacter freundii and / or Proteus mirabilis, in particular Klebsiella pneumoniae. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figures 1 to 6 They are respectively the XRPD patterns of Form A prepared according to Examples 1 to 6 of the present invention;

[0018] Figure 7 The crystal structure of Form A obtained in Example 2 of the present invention;

[0019] Figures 8 to 9 They are TGA graphs of Form A prepared according to Examples 1 and 2 of the present invention, respectively;

[0020] Fig.10 This is the DSC graph of Form A prepared according to Example 2 of the present invention;

[0021] Fig.11 is the XRPD pattern of the β-lactam compound in free acid form prepared according to Comparative Example 1;

[0022] Fig.12 is the XRPD pattern of the amorphous β-lactam compound prepared according to Comparative Example 2. DETAILED DESCRIPTION

[0023] In one aspect, the present invention provides a compound of formula (I):

[0024]

[0025] wherein n = 0-6, preferably 0.5-5, more preferably 2-3. In a specific embodiment, n is 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4 or 4.5.

[0026] Furthermore, the present invention also provides a crystalline form A of the compound of formula (I), whose X-ray powder diffraction (XRPD) Figure 2 The θ diffraction angle has characteristic diffraction peaks at 5.8±0.2°, 10.8±0.2°, 15.3±0.2°, 17.4±0.2° and 18.6±0.2°.

[0027] More preferably, the X-ray powder diffraction of the crystalline form A is Figure 2 The θ diffraction angle also has characteristic diffraction peaks at 21.3±0.2° and 36.6±0.2°.

[0028] Further preferably, the X-ray powder diffraction of the crystalline form A is Figure 2 The θ diffraction angle also has characteristic diffraction peaks at 5.1±0.2°, 11.5±0.2°, and 13.1±0.2°.

[0029] In a preferred embodiment of the present invention, the thermogravimetric analysis (TGA) graph of the crystalline form A has a weight loss of 7.4%-11.2% in the entire range of 30-170°C, preferably a weight loss of 10.3±0.5%, and also preferably a weight loss of 9.6±1.0%; more preferably, the thermogravimetric analysis graph of the crystalline form A has a weight loss of 0.5%-1.5% in the entire range of 30-53°C, preferably a weight loss of 1.0±0.3%; more preferably, the thermogravimetric analysis graph of the crystalline form A has a weight loss of 7.6%-10.0% in the entire range of 53.1-170°C, preferably a weight loss of 9.5±0.3%; also preferably, the thermogravimetric analysis graph of the crystalline form A has a weight loss of 1.0%-2.5% in the entire range of 95-170°C, preferably a weight loss of 1.5±0.2%.

[0030] In a preferred embodiment of the present invention, the differential scanning calorimetry (DSC) of the crystalline form A has an endothermic peak in the range of 70-125°C, preferably at 80±2°C and 110±2°C, and more preferably at 110±2°C.

[0031] In a preferred embodiment of the present invention, the X-ray powder diffraction pattern of the crystalline form A is substantially as follows Figure 2 shown.

[0032] On the other hand, the present invention provides a method for preparing the compound of formula (I), comprising: step (1), dissolving the compound of formula (A-1) and a sodium conversion agent in a good solvent to form a salt:

[0033]

[0034] The compound of formula (A-1) is known and can be easily obtained by a person skilled in the art or prepared according to methods disclosed in the prior art, such as the method disclosed in patent application CN113754651A, the disclosure of which is incorporated herein by reference.

[0035] In a preferred embodiment of the present invention, the good solvent is selected from at least one of methanol, water, DMSO and DMF, preferably at least one of methanol and water.

[0036] In a preferred embodiment of the present invention, the sodium conversion agent is selected from at least one of sodium lactate, sodium bicarbonate, sodium carbonate, sodium isooctanoate and sodium acetate, preferably at least one of sodium isooctanoate and sodium acetate.

[0037] In a preferred embodiment of the present invention, the amount of the good solvent is 0.5-20 mL / g of the compound of formula (A-1), preferably 1-10 mL / g of the compound of formula (A-1), and more preferably 2-6 mL / g of the compound of formula (A-1); preferably, the molar ratio of the compound of formula (A-1) to the sodium conversion agent is 1:(1-6), preferably 1:(1.5-4), and more preferably 1:(2-3.5).

[0038] In a preferred embodiment of the present invention, the temperature of step (1) is 0-35°C, preferably 2-23°C, more preferably 4-20°C, still more preferably 6-15°C or 10-20°C.

[0039] In a preferred embodiment of the present invention, the compound of formula (I) in step (1) is further crystallized. Unexpectedly, the present invention has found that the crystalline form A of the compound of formula (I) can be prepared, which further comprises: step (2), adding a poor solvent for the compound of formula (I) to the salt solution obtained in step (1) at a temperature of 0°C-30°C to form crystals.

[0040] The crystallization temperature in step (2) may be 0° C.-25° C., preferably 2-23° C., more preferably 4-20° C., still more preferably 6-15° C. or 10-20° C. Preferably, the crystallization temperature in step (2) is the same as that in step (1).

[0041] In a preferred embodiment of the present invention, in step (2), the poor solvent is selected from at least one of ethyl acetate, petroleum ether, ethanol, acetone, isopropanol and diethyl ether, preferably selected from at least one of ethyl acetate, ethanol, acetone and isopropanol.

[0042] Preferably, when the sodium transfer agent is selected from at least one of sodium octanoate and sodium acetate, the good solvent is selected from at least one of methanol and water. Also preferably, when the sodium transfer agent is selected from at least one of sodium bicarbonate and sodium carbonate, the good solvent is water.

[0043] In a preferred embodiment of the present invention, the volume ratio of the good solvent to the poor solvent may be 1:(1-20), preferably 1:(3-17), and further preferably 1:(4-12).

[0044] In a preferred embodiment of the present invention, the process further comprises step (3): filtering the crystals obtained in step (2), washing the crystals with a poor solvent and drying them.

[0045] In a preferred embodiment of the present invention, in step (3), the drying temperature is 0-80°C, preferably 15-55°C, and further preferably 30-45°C.

[0046] In another aspect, the present invention provides a pharmaceutical composition comprising a compound of formula (I) of the present invention or a compound of formula (I) prepared according to the method of the present invention and a pharmaceutically acceptable adjuvant.

[0047] In the present invention, the term "pharmaceutically acceptable adjuvant" includes, but is not limited to, at least one of arginine, sodium bicarbonate, sodium carbonate, sulbactam sodium and avibactam sodium.

[0048] In a preferred embodiment of the present invention, the pharmaceutical composition can be in any dosage form, such as tablets, pills, capsules, powders, granules or suppositories.

[0049] In the present invention, the pharmaceutical composition may also be an injection, especially a powder injection.

[0050] In another aspect, the present invention also provides the use of the compound of formula (I) or its pharmaceutical composition of the present invention for resisting Gram-negative bacteria, in particular Escherichia coli, Klebsiella pneumoniae, Klebsiella aerogenes, Enterobacter cloacae, Citrobacter freundii and / or Proteus mirabilis, in particular Klebsiella pneumoniae.

[0051] Surprisingly, the compounds of formula (I) of the present invention have improved antibacterial efficacy compared to the free acid, particularly efficacy against Klebsiella pneumoniae. Therefore, the compounds of formula (I) of the present invention or their pharmaceutical compositions are also preferably used to treat infectious diseases associated with the above-mentioned bacteria.

[0052] In addition, in the present invention, Na in the compound of formula (I) can also be replaced by K or NH4. It is also found that the crystalline form A of the compound of formula (I) of the present invention has better crystallinity, stability and / or hygroscopicity than the corresponding crystalline forms of other salts.

[0053] Example

[0054] The following examples are provided to better illustrate the present invention. It should be understood that the examples are merely exemplary and should not be construed as limiting the scope of the present invention.

[0055] In the present invention, unless otherwise stated, all temperatures are room temperature (25±2° C.) and pressures are atmospheric pressure (101 kPa).

[0056] General instructions

[0057] 1. The measurement information of X-ray powder diffraction (XRPD) is as follows:

[0058] Testing instrument: Empyrean X-ray diffractometer

[0059] Detection conditions: Cu target Kα ray, voltage 40 kV, current 40 mA, divergence slit 1 / 8°, anti-scattering slit 1 / 4°, anti-scattering slit 7.5 mm, 2θ range: 3°-60°, step size 0.02°, dwell time 40 s per step.

[0060] Testing basis: X-ray diffraction method, Part IV, 0451, 2020 edition of the Pharmacopoeia of the People's Republic of China.

[0061] 2. The measurement information of thermogravimetric analysis (TGA) is as follows:

[0062] Analytical instrument: TA Q5000; Heating rate: 10°C per minute; Protective gas: nitrogen.

[0063] 3. The measurement information of differential scanning calorimetry (DSC) is as follows:

[0064] Analytical instrument: TA Q2000; Temperature change rate: 5°C per minute; Protective gas: nitrogen.

[0065] Preparation Example

[0066] Comparative Example 1: Preparation of the compound of formula (A-1)

[0067] The compound of formula (A-1) was prepared according to the method described in Preparation Example 1 in patent application CN 113754651A. It was characterized by XRPD, as shown in Fig.11 .

[0068] Depend on Fig.11 It can be seen that the compound of formula (A-1) does not show characteristic peaks of crystals and the compound has an amorphous structure.

[0069] Example 1: Preparation of the compound of formula (I)

[0070] At 3°C, 8 g of sodium isooctanoate was dissolved in 50 mL of water, and then 10 g of the compound of formula (A-1) prepared according to Comparative Example 1 was added. At 3°C, 500 mL of acetone was added to the above mixture, and then stirred and crystallized at this temperature. After the crystallization was complete, it was filtered, washed with an appropriate amount of acetone, and vacuum dried at 40°C to obtain Form A of the compound of formula (I) with a yield of 87%. The XRPD and TGA characterizations were shown in Figure 1 and Figure 8 .

[0071] like Figure 1 As shown, the X-ray powder diffraction pattern of Form A prepared according to Example 1 of the present invention has the following characteristic peaks: characteristic diffraction peaks at 2θ of 5.8±0.2°, 10.8±0.2°, 15.3±0.2°, 17.4±0.2° and 18.6±0.2°, in addition, characteristic diffraction peaks at 21.3±0.2°, 36.6±0.2°, 5.1±0.2°, 11.5±0.2° and 13.1±0.2°.

[0072] like Figure 8 As shown, the weight loss of the crystal form A prepared in Example 1 of the present invention when heated to 170°C is 10.42%. The crystal form A loses 0.86% of its weight in the range of 30-55°C (this part is free water and solvent), and loses 9.56% of its weight in the range of 55.1-170°C (this part is crystal water). It can be concluded through calculation that the crystal form A obtained in Example 1 is disodium trihydrate, that is, n in formula (I) is 3.

[0073] Example 2

[0074] At 5°C, 4 g of sodium acetate was dissolved in 30 mL of water, and then 10 g of the compound of formula (A-1) prepared according to Comparative Example 1 was added to the above sodium acetate solution at 5°C, 350 mL of isopropanol was added to the above mixture, and then stirred for crystallization at the same temperature. After the crystallization was complete, it was filtered, washed with an appropriate amount of isopropanol, and vacuum dried at 35°C to obtain Form A of the compound of formula (I) with a yield of 94%. The crystals were characterized by XRPD, TGA and DSC, as shown in Figure 2 , Fig. 9 and Fig.10 .

[0075] like Figure 2As shown, the X-ray powder diffraction pattern of Form A prepared according to Example 2 of the present invention has the following characteristic peaks: characteristic diffraction peaks at 2θ of 5.8±0.2°, 10.8±0.2°, 15.3±0.2°, 17.4±0.2° and 18.6±0.2°, in addition, characteristic diffraction peaks at 21.3±0.2°, 36.6±0.2°, 5.1±0.2°, 11.5±0.2° and 13.1±0.2°.

[0076] like Fig. 9 As shown, the weight loss of the crystal form A prepared in Example 2 of the present invention when heated to 170°C is 10.72%. The crystal form A loses 1.03% of its weight in the range of 30-53°C (this part is free water and solvent), and loses 9.67% of its weight in the range of 53.1-170°C (this part is crystal water). It can be concluded through calculation that the crystal form A obtained in Example 2 is disodium trihydrate, that is, n in formula (I) is 3.

[0077] like Fig.10 As shown, the crystalline form A prepared according to Example 2 of the present invention has endothermic peaks at 79.9°C and 110.3°C.

[0078] In addition, the single crystal structure analysis of the crystal form A obtained in Example 2 showed that the crystal form A had a monoclinic P21 space group, the unit cell parameters were listed in Table 1 below, and the single crystal structure was shown in Figure 7 .

[0079] Table 1 Unit cell parameters

[0080]

[0081] Example 3

[0082] At 5°C, 10 g of the compound of formula (A-1) prepared according to comparative example 1 was dissolved in 20 mL of water, and then 4 g of sodium carbonate was added; 100 mL of ethanol was added to the above mixture at 5°C, and then stirred at this temperature for crystallization. After the crystallization was complete, it was filtered, washed with an appropriate amount of ethanol, and vacuum dried at 45°C to obtain the crystalline form A of the compound of formula (I) with a yield of 87%. Characterized by XRPD, shown in Figure 3 .

[0083] like Figure 3 As shown, the X-ray powder diffraction pattern of Form A prepared according to Example 3 of the present invention has the following characteristic peaks: characteristic diffraction peaks at 2θ of 5.8±0.2°, 10.8±0.2°, 15.3±0.2°, 17.4±0.2° and 18.6±0.2°, in addition, characteristic diffraction peaks at 21.3±0.2°, 36.6±0.2°, 5.1±0.2°, 11.5±0.2° and 13.1±0.2°.

[0084] Example 4

[0085] At 15°C, 10 g of the compound of formula (A-1) prepared according to comparative example 1 was dissolved in 30 mL of methanol and water (volume ratio 1:1), and then 7 g of sodium isooctanoate was added; 500 mL of acetone was added to the mixture at 15°C, and then stirred and crystallized at the same temperature. After the crystallization was complete, it was filtered, washed with an appropriate amount of acetone, and vacuum dried at 40°C to obtain Form A of the compound of formula (I) with a yield of 89%. Characterized by XRPD, shown in Figure 4 .

[0086] like Figure 4 As shown, the X-ray powder diffraction pattern of Form A prepared according to Example 4 of the present invention has the following characteristic peaks: characteristic diffraction peaks at 2θ of 5.8±0.2°, 10.8±0.2°, 15.3±0.2°, 17.4±0.2° and 18.6±0.2°, in addition, characteristic diffraction peaks at 21.3±0.2°, 36.6±0.2°, 5.1±0.2°, 11.5±0.2° and 13.1±0.2°.

[0087] Example 5

[0088] At 12°C, 6 g of sodium acetate was dissolved in 20 mL of water, and then 10 g of the compound of formula (A-1) prepared according to Comparative Example 1 was added. At 12°C, 300 mL of isopropanol was added to the above mixture, and then stirred at this temperature for crystallization. After the crystallization was complete, it was filtered, washed with an appropriate amount of isopropanol, and vacuum dried at 30°C to obtain Form A of the compound of formula (I) with a yield of 92%. Characterized by XRPD, shown in Figure 5 .

[0089] like Figure 5 As shown, the X-ray powder diffraction pattern of Form A prepared according to Example 5 of the present invention has the following characteristic peaks: characteristic diffraction peaks at 2θ of 5.8±0.2°, 10.8±0.2°, 15.3±0.2°, 17.4±0.2° and 18.6±0.2°, in addition, characteristic diffraction peaks at 21.3±0.2°, 36.6±0.2°, 5.1±0.2°, 11.5±0.2° and 13.1±0.2°.

[0090] Example 6

[0091] At 23°C, 10 g of the compound of formula (A-1) prepared according to comparative example 1 was dissolved in 30 mL of water, and then 4 g of sodium bicarbonate was added; 360 mL of ethanol was added to the above mixture at 23°C, and then stirred at the same temperature for crystallization. After the crystallization was complete, it was filtered, washed with an appropriate amount of ethanol, and vacuum dried at 35°C to obtain the crystalline form A of the compound of formula (I) with a yield of 89%. Characterized by XRPD, shown in Figure 6 .

[0092] like Figure 6 As shown, the X-ray powder diffraction pattern of Form A prepared according to Example 6 of the present invention has the following characteristic peaks: characteristic diffraction peaks at 2θ of 5.8±0.2°, 10.8±0.2°, 15.3±0.2°, 17.4±0.2° and 18.6±0.2°, in addition, characteristic diffraction peaks at 21.3±0.2°, 36.6±0.2°, 5.1±0.2°, 11.5±0.2° and 13.1±0.2°.

[0093] Comparative Example 2

[0094] 1 g of the crystalline form A of the compound of formula (I) obtained in Example 1 was dissolved in 10 mL of water and then freeze-dried to obtain an amorphous solid of the compound of formula (I). Characterized by XRPD, the results are shown in Fig.12 .

[0095] Depend on Fig.12 It can be seen that the obtained compound does not show characteristic peaks of crystals and has an amorphous structure.

[0096] Effect Example

[0097] 1. In vitro efficacy test

[0098] In order to evaluate the antibacterial activity of the compound of formula (I) (prepared by Example 2), an in vitro antibacterial activity study was conducted against Aztreonam, Ceftazidime / avibactam, Cefepime, Amikacin, Ciprofloxacin, Colistin, Meropenem, Ertapenem and Tegecycline.

[0099] Minimum inhibitory concentration (MIC) determination

[0100] According to the CLSI standard, the agar dilution method was used for drug sensitivity test. The test bacteria were cultured with appropriate liquid or solid culture medium, and the drug was diluted twice with broth culture medium to various required concentrations, and then added to the plate in appropriate amounts. After the agar culture medium was melted, it was quantitatively injected into the plate containing the drug solution and mixed, so that the final concentration of the drug in the plate was 128, 64, 32, 16, 8, 4, 2, 1, 0.5, 0.25, 0.12, 0.06, and 0.03 μg / mL, respectively. The test bacteria were inoculated with a multi-point inoculator, and the inoculation amount was about 104 CFU / point. The culture was placed at a constant temperature of 35°C, and the results were observed after 18 hours. The minimum concentration of the drug contained in the plate with sterile growth was the minimum inhibitory concentration (MIC). The results are listed in Tables 2 and 3 below.

[0101] Table 2-1 MIC determination of the compound of formula (I) and its control drug against clinical isolated bacteria (μg / mL)

[0102]

[0103]

[0104] Note: CRE stands for carbapenem-resistant Enterobacteriaceae

[0105] Table 2-2 MIC determination of the compound of formula (I) and its control drug against clinical isolated bacteria (μg / mL)

[0106]

[0107] Table 3-1 MIC determination of the compound of formula (I) and its control drug against carbapenem-resistant strains (μg / mL)

[0108]

[0109]

[0110] Note: CRECL stands for carbapenem-resistant Enterobacter cloacae, CREC stands for carbapenem-resistant Escherichia coli

[0111] Table 3-2 MIC determination of the compound of formula (I) and its control drug against ESBLs-producing strains (μg / mL)

[0112]

[0113] Note: ESBLs stands for β-lactamase

[0114] The test results show that the compound of formula (I) has a strong antibacterial effect on a variety of common clinical Gram-negative bacteria (including ESBLs-producing strains and carbapenem-resistant strains), and has a strong antibacterial effect on Escherichia coli, Klebsiella pneumoniae, Klebsiella aerogenes, Enterobacter cloacae, Citrobacter freundii and Proteus mirabilis.

[0115] The compound of formula (I) has strong antibacterial activity against 150 strains of Escherichia coli, with MIC50 and MIC90 values ​​of 0.12 and 2 μg / mL, respectively. The antibacterial activity of the compound of formula (I) is 2 times that of meropenem, 4 times that of amikacin, and 8 times that of ertapenem, which is significantly better than aztreonam, ciprofloxacin, and cefepime.

[0116] The compound of formula (I) has strong antibacterial activity against 45 strains of CREC, with MIC50 and MIC90 values ​​of 1 and 32 μg / mL, respectively. The antibacterial activity of the compound of formula (I) is similar to that of meropenem, twice that of ertapenem, and superior to aztreonam, ceftazidime / avibactam, cefepime, amikacin and ciprofloxacin.

[0117] The compound of formula (I) has strong antibacterial activity against 57 strains of ESBLs-producing Escherichia coli (non-CREC strains), with MIC50 and MIC90 values ​​of 0.12 and 0.5 μg / mL, respectively. The antibacterial activity of the compound of formula (I) is similar to that of colistin and tigecycline, 8 times that of amikacin, and significantly better than aztreonam, cefepime and ciprofloxacin.

[0118] The compound of formula (I) has strong antibacterial activity against 167 strains of Klebsiella pneumoniae, with MIC50 and MIC90 values ​​of 0.12 and 4 μg / mL, respectively. The antibacterial activity of the compound of formula (I) is 8 times that of amikacin, 16 times that of cefepime, ciprofloxacin and meropenem, better than aztreonam and ertapenem, and slightly weaker than tigecycline.

[0119] The compound of formula (I) has strong antibacterial activity against 56 strains of CRKP, with MIC50 and MIC90 values ​​of 2 and 32 μg / mL, respectively. The antibacterial activity of the compound of formula (I) is 2 times that of meropenem, 4 times that of cefepime and ciprofloxacin, and is superior to aztreonam, amikacin and ertapenem.

[0120] The compound of formula (I) has strong antibacterial activity against 30 strains of ESBLs-producing Klebsiella pneumoniae (non-CRKP strains), with MIC50 and MIC90 values ​​of 0.5 and 16 μg / mL, respectively. The antibacterial activity of the compound of formula (I) is 2 times that of cefepime, 4 times that of ciprofloxacin, and 8 times that of aztreonam.

[0121] The compound of formula (I) has strong antibacterial activity against 55 strains of Enterobacter cloacae, with MIC50 and MIC90 values ​​of 4 and 32 μg / mL, respectively. The antibacterial activity of the compound of formula (I) is similar to that of ertapenem, 4 times that of ciprofloxacin, and superior to aztreonam, ceftazidime / avibactam, cefepime and amikacin.

[0122] The compound of formula (I) has strong antibacterial activity against 39 strains of CRECL, with MIC50 and MIC90 values ​​of 8 and 32 μg / mL, respectively. The antibacterial activity of the compound of formula (I) is similar to that of ertapenem, 4 times that of ciprofloxacin, and superior to aztreonam, ceftazidime / avibactam, cefepime and amikacin.

[0123] The compound of formula (I) has strong antibacterial activity against 36 strains of Enterobacter aerogenes, with MIC50 and MIC90 values ​​of 0.12 and 0.25 μg / mL, respectively. The antibacterial activity of the compound of formula (I) is similar to that of ceftazidime / avibactam and ertapenem, 2 times that of colistin and tigecycline, 8 times that of amikacin, and 16 times that of aztreonam.

[0124] The compound of formula (I) has strong antibacterial activity against 37 strains of Proteus mirabilis, with MIC50 and MIC90 values ​​of ≤0.03 and ≤0.03 μg / mL, respectively. Its antibacterial activity is similar to that of aztreonam / avibactam and ertapenem, superior to aztreonam, ceftazidime / avibactam and meropenem, and significantly superior to cefepime, amikacin, ciprofloxacin, colistin and tigecycline.

[0125] 2. Antibacterial activity comparison experiment

[0126] The antibacterial activity results of the compound of formula (A-1) prepared according to Comparative Example 1 and the compound of formula (I) prepared according to Example 2 against Klebsiella pneumoniae are listed in the following Table 4. The minimum inhibitory concentration MIC was determined as described above.

[0127] Table 4 Antibacterial activity results

[0128]

[0129] The test results show that compared with the compound of formula (A-1), the compound of formula (I) of the present invention has a better anti-Klebsiella pneumoniae effect.

[0130] 2. Solubility test

[0131] The solubility of the compound of formula (A-1) prepared according to Comparative Example 1, the crystalline form A of the compound of formula (I) prepared according to Example 2, and the amorphous form of the compound of formula (I) prepared according to Comparative Example 2 were measured at room temperature. The test results are listed in the following Table 5:

[0132] Table 5 Solubility data

[0133]

[0134] As can be seen from Table 2, the crystalline form A of the compound of formula (I) of the present invention can achieve improved solubility even without using a co-solvent. Unexpectedly, the crystalline form A of the present invention shows higher solubility than its amorphous form.

[0135] 3. Stability

[0136] Weigh 6 parts of 1 g of the crystalline form A of the compound of formula (I) prepared according to Example 2 of the present invention and 6 parts of 1 g of the amorphous form of the compound of formula (I) prepared according to Comparative Example 2 and store them in a constant temperature and humidity test box at 25°C / 60%RH, and test their content, pH value and moisture after the intervals shown in Table 6 (month refers to a natural month). The test results are summarized in Table 6, where the active content refers to the content of the compound of formula (I), in weight %.

[0137] Table 6 Stability data

[0138]

[0139] As can be seen from the table above, after the crystal form A of the present invention is stored for up to 12 months at 25°C / 60%RH, its quality remains basically unchanged, which indicates that it has high stability. In contrast, the corresponding amorphous compound shows lower stability.

[0140] Although several preferred embodiments of the present invention have been described for illustrative purposes, those skilled in the art will appreciate that various modifications, additions and substitutions are possible, without departing from the scope and spirit of the invention as disclosed in the accompanying claims.

Claims

1. Compound of formula (I): wherein n=0-6, preferably 2-3.

2. The compound according to claim 1, which is in form A, and the X-ray powder diffraction pattern of the form A has characteristic diffraction peaks at 2θ diffraction angles of 5.8±0.2°, 10.8±0.2°, 15.3±0.2°, 17.4±0.2° and 18.6±0.2°. Preferably, the X-ray powder diffraction pattern of the form A also has characteristic diffraction peaks at 21.3±0.2° and 36.6±0.2°. More preferably, the X-ray powder diffraction pattern of the form A also has characteristic diffraction peaks at 2θ diffraction angles of 5.1±0.2°, 11.5±0.2° and 13.1±0.2°.

3. The compound according to any one of claims 1 to 3, which has an X-ray powder diffraction pattern as shown in Figure 2.

4. A compound according to any one of claims 1 to 4, having a monoclinic P21 space group and the following unit cell parameters: α＝90° β＝107.352(5)° γ = 90°.

5. A method for preparing a compound of formula (I) as described in any one of claims 1 to 4, comprising: The compound of formula (A-1) and the sodium conversion agent are dissolved in a good solvent to form a salt, Wherein, the good solvent is preferably selected from at least one of methanol, water, DMSO and DMF, more preferably at least one of methanol and water; The sodium conversion agent is preferably selected from at least one of sodium lactate, sodium bicarbonate, sodium carbonate, sodium octanoate and sodium acetate, more preferably at least one of sodium octanoate and sodium acetate; The amount of the good solvent used is preferably 0.5-20 mL / g of the compound of formula (A-1), more preferably 1-10 mL / g of the compound of formula (A-1), and most preferably 2-6 mL / g of the compound of formula (A-1).

6. The method according to claim 5, further comprising: Add a poor solvent for the compound of formula (I) to the salt solution at a temperature of 0°C to 30°C to form crystals. The poor solvent is preferably selected from at least one of ethyl acetate, petroleum ether, ethanol, acetone, isopropanol and ether, more preferably at least one of ethyl acetate, ethanol, acetone and isopropanol. The volume ratio of the good solvent to the poor solvent is preferably 1:(1-20), more preferably 1:(3-17).

7. A pharmaceutical composition, characterized in that It comprises a compound of formula (I) according to any one of claims 1 to 4 or a compound of formula (I) prepared by the method described in claim 5 or 6, and a pharmaceutically acceptable auxiliary agent.

8. The pharmaceutical composition according to claim 7, which is in the form of tablets, pills, capsules, powders, granules, suppositories or injections, particularly in the form of powder injections.

9. A compound of formula (I) according to any one of claims 1 to 4 or a pharmaceutical composition according to claim 6 or 7 for use against Gram-negative bacteria, in particular Escherichia coli, Klebsiella pneumoniae, Klebsiella aerogenes, Enterobacter cloacae, Citrobacter freundii and / or Proteus mirabilis.

10. The use according to claim 10 for the treatment of infectious diseases associated with Gram-negative bacteria, in particular Escherichia coli, Klebsiella pneumoniae, Klebsiella aerogenes, Enterobacter cloacae, Citrobacter freundii and / or Proteus mirabilis.

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