Beta-lactamase inhibitor intermolecular co-crystal or salt and preparation method and application thereof

Abstract

The present application relates to a beta-lactamase inhibitor intermediate co-crystal or salt, a preparation method and application thereof, and belongs to the technical field of pharmaceutical chemical engineering. The structural formula is as follows: R2 represents benzyl or allyl; R3 represents an alkylamine group with a protective group, a cycloalkylamine group, an aromatic amine group, and a derivative amine group. The preparation method comprises the following steps: dissolving compound I in a solvent A, adding a base solution to react under the condition of less than 0℃, then adding an acid to adjust the pH to be acidic, adding a solvent B to extract the product, separating the layers, and leaving the organic phase; and then adding a beta-lactamase inhibitor side chain R3H to react. The beta-lactamase inhibitor is prepared by using the intermediate co-crystal or the new salt, the purity is greater than 99.90%, the pharmaceutical quality standard of the crude drug is met, the process operation is safe, simple, convenient, green and environment-friendly; the atom economy, the selectivity, the purity and the yield of each step reaction are high, and the production can be enlarged.

Description

Technical Field

[0001] This invention relates to a novel β-lactamase inhibitor intermediate cocrystal or new salt compound, its preparation method and uses, belonging to the field of pharmaceutical and chemical technology. Background Technology

[0002] Second-generation β-lactamase inhibitors refer to β-lactamase inhibitors with novel structural types. Their chemical structures do not contain a β-lactam ring; instead, they use diazabicyclooctanone as a backbone structure. These inhibitors inhibit β-lactamases, restoring antibiotic activity. The structures of these β-lactamase inhibitors are mainly composed of diazabicyclooctanone and side chains, and include retelebactam, zidebactam, nacubactam, WCK-5153, FPI-1465, and WCK-6395.

[0003] .

[0004] Taking relebactam as an example, this patent illustrates its research background, highlighting the similarity in the preparation methods of such compounds. Relebactam's chemical name is mono[(1R,2S,5R)-7-oxo-2-[(4-piperidinylamino)carbonyl]-1,6-diazabicyclo[3.2.1]oct-6-yl] ester. It belongs to the diazabicyclooctane inhibitor class and possesses broad-spectrum anti-β-lactamase activity, including class A (extended-spectrum β-lactamases and KPC enzymes) and class C (AmpC enzymes).

[0005] The literature reports the following main methods for preparing relebactam:

[0006] Option 1. Patent documents WO2009091856A1 and Org. Lett. 2011, 13, 20, 5480–5483: This route uses expensive N-Cbz-aminopiperidine 1 as the starting material and uses a heavy metal iridium catalyst; it uses toxic reagents such as sulfonyl chloride, methanesulfonic acid, and pyridine; this route involves 13 steps of reaction, with an overall yield of 10%, poor operating environment, large amounts of waste salt and wastewater, poor economic efficiency, and is not conducive to industrial production.

[0007] .

[0008] Scheme 2. Patent documents WO2014200786A1 and Org. Lett. 2014, 16, 174−177: This route uses enantiomer 13, which is difficult to obtain and has low purity, as the starting material, avoiding the use of expensive iridium catalysts. The total route is shortened to 11 steps, with a yield of 42%. However, it still uses toxic reagents such as sulfonyl chloride and methanesulfonic acid, which are not suitable for industrial scale-up.

[0009] .

[0010] Scheme 3. Reference: Chem. Commun., 2022, 58, 10869-10872: This route uses commercially available 4-hydroxy-2-butanone as the starting material and employs a rhodium catalyst for asymmetric hydrogenation to construct a chiral center. The process involves first constructing a chiral piperidine ring 27, then amide condensing it to an aminopiperidine side chain to obtain 22, and finally synthesizing relebactam using Merck's synthetic method. The total reaction involves 16 steps with an overall yield of 15%. This route is lengthy and involves many steps, requiring the construction of a six-membered ring and two chiral centers, and utilizes a heavy metal rhodium catalyst. While suitable for laboratory research, it is not ideal for industrial-scale production.

[0011] . Summary of the Invention

[0012] To address the shortcomings of existing technologies, this invention discloses a co-crystal or new salt compound (II) of β-lactamase inhibitor intermediates, its preparation method, and its uses. β-lactamase inhibitors are synthesized using this intermediate (II), yielding relebactam, zidebactam, nacubactam, WCK-5153, FPI-1465, WCK-6395, etc., with a purity greater than 99.90%. This process is simple to operate, uses readily available raw materials, has low cost, and produces high-purity products, which is beneficial for industrial-scale production.

[0013] The cocrystal or salt of the β-lactamase inhibitor intermediate has the following structural formula:

[0014]

[0015] R2 represents benzyl or allyl;

[0016] R3 represents a protecting alkylamine group, cycloalkylamine group, aromatic amine group, or their derivative amine group, selected from... .

[0017] P represents the protecting group of tert-butoxycarbonyl (Boc), benzyloxycarbonyl (Cbz), methoxycarbonyl (Fmoc), and allyloxycarbonyl (Alloc).

[0018] The specific synthesis process route is shown below:

[0019] .

[0020] The synthesis technology solution of this invention is as follows:

[0021] Using compound I, compound I is dissolved in solvent A, and an alkaline solution is added to react under conditions below 0°C (preferably -5°C to -20°C). Then, acid is added to adjust the pH to acidic, and solvent B is added to extract the product, resulting in separation of layers and retaining the organic phase. Then, the side chain R3H of the β-lactamase inhibitor is added to react. After the reaction is completed, a white solid or crystal precipitates, which is filtered to obtain the β-lactamase inhibitor intermediate cocrystal II or the new salt II.

[0022] In compound I, R1 represents an alkoxy group, selected from methoxy, ethoxy, propoxy, butoxy, or benzyloxy, etc. Further, compound I is selected from one or more of the following: (2S,5R)-6-(benzyloxy)-7-oxo-1,6-diazabicyclo[3.2.1]octane-2-carboxylic acid ethyl ester, (2S,5R)-6-(benzyloxy)-7-oxo-1,6-diazabicyclo[3.2.1]octane-2-carboxylic acid methyl ester, or (2S,5R)-6-(benzyloxy)-7-oxo-1,6-diazabicyclo[3.2.1]octane-2-carboxylic acid benzyl ester.

[0023] R3H is an alkylamine, cycloalkylamine, aromatic amine, or its derivative containing a protecting group. R3H is selected from... P represents a protecting group of tert-butoxycarbonyl (Boc), benzyloxycarbonyl (Cbz), methoxycarbonyl (Fmoc), or allyloxycarbonyl (Alloc).

[0024] Preferably, the molar ratio of compound I to the β-lactamase inhibitor side chain R3H in the above steps is 1:0.8~1.5.

[0025] Preferably, solvent A is one or more of acetone, methanol, ethanol, isopropanol, tetrahydrofuran, acetonitrile, etc., and the addition of acid and / or base includes water; solvent B is one or more of dichloromethane, chloroform, ethyl acetate, butyl acetate, isopropyl acetate, 1,2-dichloroethane, toluene, 2-methyltetrahydrofuran, cyclopentyl methyl ether, chlorobenzene, etc.; base is selected from sodium hydroxide, lithium hydroxide, potassium hydroxide, etc., with sodium hydroxide being preferred; acid is selected from hydrochloric acid, sulfuric acid, nitric acid, acetic acid, hydrobromic acid, hydroiodic acid, phosphoric acid, carbonic acid, hypochlorous acid, etc., with hydrochloric acid being preferred.

[0026] Further β-lactamase inhibitor intermediate cocrystals or salts are selected from one or more of the following:

[0027] .

[0028] The method for preparing β-lactamase inhibitors using the obtained β-lactamase inhibitor intermediate cocrystal II or new salt II is further described.

[0029] .

[0030] Specifically, the steps include the following:

[0031] (1) Dissolve the β-lactamase inhibitor intermediate cocrystal II or new salt II in solvent C, add condensing agent, and after the reaction is complete, add solvent B' for extraction, separate the layers, retain the organic phase, wash with inorganic alkaline aqueous solution, rotary evaporate, crystallize, and obtain compound III.

[0032] (2) Compound III is prepared by de-R2 reaction and sulfonation reaction in solvent D under catalytic conditions, followed by ammonium salt formation reaction;

[0033] (3) In solvent E or without solvent, the compound of formula IV is deprotected by the protecting group P in R3 and then crystallized by adding a crystallization solvent to prepare β-lactamase inhibitors, including retribactam, Zidebactam, WCK-5153, Nacubactam, FPI-1465, and WCK-6395.

[0034] Preferably, in step (1), solvent C is an organic solvent or an organic solvent and water, and the organic solvent is selected from one or more of acetone, methanol, ethanol, isopropanol, tetrahydrofuran, acetonitrile, etc.; solvent B' is an organic solvent or an organic solvent and water, and the organic solvent is selected from one or more of dichloromethane, chloroform, ethyl acetate, butyl acetate, isopropyl acetate, 1,2-dichloroethane, toluene, 2-methyltetrahydrofuran, cyclopentyl methyl ether, chlorobenzene, etc.; at least one of solvent C and solvent B' contains water; in step (2), solvent D is an organic solvent or an organic solvent and water, and the organic solvent is selected from one or more of acetone, methanol, ethanol, isopropanol, tetrahydrofuran, acetonitrile, dichloromethane, chloroform, ethyl acetate, etc., and also includes organic bases, etc.; in step (3), solvent E is dichloromethane, chloroform, ethyl acetate, 2-methyltetrahydrofuran, methyl tert-butyl ether, etc.

[0035] Preferably, the condensing agent is 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI), 1-hydroxybenzotriazole (HOBt), N,N'-dicyclohexylcarbodiimide (DCC), N,N'-diisopropylcarbodiimide (DIC), 1-hydroxy-7-azabenzotriazole (HOAt), benzotriazole-N,N,N',N'-tetramethylurea hexafluorophosphate (HBTU), or 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (HATU). One or more of benzotriazine-1-yl-oxy-tris(dimethylamino)phosphine hexafluorophosphate (BOP), (benzotriazine-1-yl-oxy)tripyrrolidinylphosphine hexafluorophosphate (PyBOP), N,N'-carbonyldiimidazole (CDI), propylphosphoric anhydride (T3P), triethylamine (TEA), N,N-diisopropylethylamine (DIPEA), pyridine, 4-dimethylaminopyridine (DMAP) or 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), preferably a combination of EDCI and HOBt.

[0036] Preferably, in step (2), when R2 is benzyl or allyl, the removal of R2 is preferably hydrogen and palladium on carbon; the sulfonation is selected from trimethylamine sulfur trioxide, triethylamine sulfur trioxide or pyridine sulfur trioxide complex; the ammonium salt formation reaction is preferably tetrabutylammonium acetate or tetrabutylammonium hydrogen sulfate.

[0037] Preferably, in step (3), under nitrogen protection, when the protecting group P is Boc, the deBoc reagent is preferably trifluoroacetic acid; when the protecting group P is benzyloxycarbonyl (Cbz), the debenzylioxycarbonyl (Cbz) reagent is preferably hydrogen bromide acetic acid solution; when the protecting group P is methoxycarbonyl (Fmoc), the demethoxycarbonyl (Fmoc) reagent is preferably 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU); when the protecting group P is allyloxycarbonyl (Alloc), the deallyoxycarbonyl (Alloc) reagent is preferably tetra(triphenylphosphine)palladium.

[0038] The crystallization solvent is selected from methyl tert-butyl ether.

[0039] The technical features and beneficial effects of this invention are as follows:

[0040] 1. This invention uses (2S,5R)-6-(benzyloxy)-7-oxo-1,6-diazabicyclo[3.2.1]octane-2-carboxylic acid ester as starting material. This material can be prepared according to existing technology. The raw material is inexpensive and readily available. It does not require expensive protective agents, metal catalysts and carbonylation reagents. The cost is low, the process is safe and simple to operate, and it is green and environmentally friendly. Each step of the reaction has high atom economy, high selectivity, high purity and high yield, and is suitable for large-scale production.

[0041] 2. The β-lactamase inhibitor intermediate cocrystal or new salt compound II has the characteristics of large particle size, easy filtration, non-hygroscopicity and chemical stability, avoiding the deterioration problem caused by storing carboxyl and amino compounds alone; at the same time, the cocrystal or new salt compound II is the carboxyl and amino raw material for preparing intermediate III, with a molar ratio of 1:1, which makes feeding simple and more precise.

[0042] 3. Using the method of this invention, not only can levobactam or its intermediate cocrystal II or new salt II compounds be prepared, but also a series of diazabicyclooctanone (DBOs) β-lactamase inhibitor products or intermediates, such as Zidebactam, Nacubactam, WCK-5153, FPI-1465, WCK-6395, etc., can be prepared using different β-lactamase inhibitor side chains R3H. Furthermore, the yields are high, and the purity is high (e.g., greater than 99.90%), meeting the pharmaceutical quality standards for active pharmaceutical ingredients. The process is safe, simple, and environmentally friendly; each reaction step exhibits high atom economy, high selectivity, high purity, and high yield, making it suitable for scale-up production. Attached Figure Description

[0043] Figure 1 For compound II-01 1 H NMR;

[0044] Figure 2 For compound II-01 13 C NMR;

[0045] Figure 3 The HPLC chromatogram of compound II-01 is shown below.

[0046] Figure 4 The image shows the XRD pattern of compound II-01. Detailed Implementation

[0047] The present invention has been described in detail below with reference to embodiments, but the present invention is not limited thereto.

[0048] Example 1:

[0049]

[0050] (1) Intermediate II-01: 1-(tert-Butoxycarbonyl)piperidine-4-amino-(2S,5R)-6-(benzyloxy)-7-oxo-1,6-diazabicyclo[3.2.1]octane-2-carboxylate

[0051] At room temperature, 50.00 g of (2S,5R)-6-(benzyloxy)-7-oxo-1,6-diazabicyclo[3.2.1]octane-2-carboxylic acid ethyl ester I-01 and acetone (250 mL) were added to a flask. The flask was then heated to -15°C externally and -10±5°C internally. A solution of sodium hydroxide (7.229 g) in water (400 mL) was added dropwise. After the addition was complete, the reaction was detected by TLC or HPLC to confirm its completeness. The flask was then cooled to room temperature, and 600 mL of isopropyl acetate was added. The pH was adjusted to 1-3 by adding 1 M hydrochloric acid (approximately 200 mL), and the mixture was stirred. After the reaction was complete, the mixture separated into layers, and the organic phase was retained. A solution of 32.903 g of β-lactamase inhibitor 1-Boc-4-aminopiperidine in isopropyl acetate (250 mL) was added, and the mixture was stirred at room temperature for 1 h. The mixture was then filtered, dried, and the product was 75.3 g. g of white powder, which is intermediate II-01, with a molar yield of 96.2% and a purity of 99.811%.

[0052] Alternatively, at room temperature, (2S,5R)-6-(benzyloxy)-7-oxo-1,6-diazabicyclo[3.2.1]octane-2-carboxylic acid ethyl ester I-01 (50.00 g) and acetone (250 mL) were added to a flask, and the mixture was transferred to an external temperature of -15°C and an internal temperature of -10±5°C. A solution of sodium hydroxide (7.229 g) in water (400 mL) was added dropwise. After the addition was complete, the reaction was detected by TLC or HPLC to confirm its completeness. The mixture was then transferred to room temperature, and dichloromethane (500 mL) was added. The pH was adjusted to 1-3 by adding 1 M hydrochloric acid (approximately 200 mL). The mixture was allowed to separate into layers, and the organic phase was retained. A solution of the β-lactamase inhibitor 1-Boc-4-aminopiperidine (32.903 g) in dichloromethane (100 mL) was added, and the mixture was stirred at 10°C for 1 h. The mixture was then filtered, dried, and the solution was 74.1 g. g of white powder, which is intermediate II-01, with a molar yield of 94.6% and a purity of 99.912%.

[0053] (2) Intermediate III-01: 4-[[[(1R,2S,5R)-7-oxo-6-(phenylmethoxy)-1,6-diazabicyclo[3.2.1]octane-2-yl]carbonyl]amino]-1-piperidinecarboxylic acid tert-butyl ester

[0054] At room temperature, II-01 (70.00 g), acetonitrile (168 mL), and tap water (42 mL) were added to a flask; EDCI (42.236 g), HOBT (33.742 g) in acetonitrile (108 mL), and tap water (32 mL) were added, and the reaction was monitored by TLC or HPLC until complete; isopropyl acetate (521 mL) and tap water (420 mL) were added to the system, and the mixture was stirred for 10 min, separating the layers, and retaining the organic phase. The organic phase was washed twice with sodium bicarbonate (12.3 g) in water (120 mL); the organic phase was evaporated to dryness, and acetone (168 mL) and water (840 mL) were added and stirred for 1 h. The mixture was filtered and dried to obtain 63.5 g of white powder, which was intermediate III-01, 94.3%, with a purity of 99.363%.

[0055] Alternatively, at room temperature, add II-01 (70.00 g), acetonitrile (168 mL), triethylamine (29.7 mL), and tap water (42 mL) to a flask; add EDCI (42.236 g), HOBT (33.742 g) in acetonitrile (108 mL), and tap water (32 mL), and check the reaction for completeness by TLC or HPLC; add isopropyl acetate (521 mL) and tap water (420 mL) to the system, stir for 10 min, separate the layers, and retain the organic phase. Wash the organic phase twice with a solution of potassium dihydrogen phosphate (9.994 g) and dipotassium hydrogen phosphate trihydrate (13.409 g) in water (210 mL); evaporate the organic phase to dryness, add acetone (168 mL) and water (840 mL), and stir for 1 h. After filtration and drying, 64.72 g of white powder was obtained, which was intermediate III-01, 96.1%, with a purity of 99.563%.

[0056] (3) Intermediate IV-01: [(1R,2S,5R)-2-([1-(tert-butyloxycarbonyl)piperidin-4-yl]aminocarbonyl)-7-oxo-1,6-diazabicyclo[3.2.1]octane-6-yl]tetrabutylammonium sulfate

[0057] At room temperature, III-01 (60.00 g), trimethylamine sulfur trioxide (26.040 g), 10% Pd / C (1.5 g), triethylamine (3.386 mL), isopropanol (240 mL), and water (240 mL) were added to a flask; the mixture was purged with nitrogen three times and hydrogen three times, and the reaction was detected to be complete by TLC or HPLC; palladium on carbon was removed by filtration, and the filtrate was washed once with isopropyl acetate (300 mL), retaining the aqueous phase; sodium bicarbonate (16.928 g) and tetrabutylammonium bisulfate (49.757 g) were added to the aqueous phase, followed by extraction with dichloromethane (420 mL), retaining the dichloromethane phase, which was then evaporated to dryness to obtain a pale yellow oil, which was intermediate IV-01 with a purity >98%. Based on a 100% yield, the reaction was directly carried out in the next step.

[0058] (4) Relebactam

[0059] At -10±5℃, the pale yellow oily substance IV-01 (approximately 90.28 g) and dichloromethane (90 mL) from the previous step were added to a flask. Under nitrogen protection, trifluoroacetic acid (230.45 mL) was added dropwise. The reaction was detected by TLC or HPLC to ensure complete reaction. The system was then transferred to room temperature, and methyl tert-butyl ether (1800 mL) was added, resulting in the precipitation of a white solid. The solid was filtered to obtain crude white solid relebactam. The crude white solid relebactam was suspended in ethanol (600 mL) and water (90 mL), and the pH was adjusted to 6-7 by adding an ethanol solution of sodium isooctanoate. Then, ethanol (400 mL) was added, and the mixture was stirred for 1 h. The mixture was filtered and dried to obtain 38.2 g of white powder, which was relebactam, with a molar yield of 83.8% and a purity of 99.918%.

[0060] Example 2:

[0061]

[0062] (1) Intermediate II-02: (3R)-3-(hydrazylcarbonyl)piperidine-1-carboxylate tert-butyl ester-(2S,5R)-6-(benzyloxy)-7-oxo-1,6-diazabicyclo[3.2.1]octane-2-carboxylate

[0063] At room temperature, 50.00 g of (2S,5R)-6-(benzyloxy)-7-oxo-1,6-diazabicyclo[3.2.1]octane-2-carboxylic acid methyl ester I-02 and acetone (250 mL) were added to a flask. The flask was then heated to -15°C externally and -10±5°C internally. A 400 mL solution of sodium hydroxide (7.578 g) in water was added dropwise. After the addition was complete, the reaction was detected by TLC or HPLC to confirm its completeness. The flask was then cooled to room temperature, and 400 mL of dichloromethane was added. The pH was adjusted to 1-3 with an aqueous solution of phosphoric acid. The mixture was separated into layers, and the organic phase was retained. A 250 mL solution of isopropyl acetate containing 41.903 g of tert-butyl piperidine-1-carboxylate, a β-lactamase inhibitor, was added. The mixture was stirred at room temperature for 1 h, filtered, and dried to obtain 83.6 g of carboxylic acid. g of white powder, namely intermediate II-02, with a molar yield of 93.4% and a purity of 99.237%.

[0064] (2) Intermediate III-02: (2S,5R)-6-benzyloxy-7-oxo-2-[[(3R)-1-(tert-butoxycarbonyl)piperidine-3-formyl]hydrazine carbonyl]-1,6-diazabicyclo[3.2.1]octane

[0065] At room temperature, 70.00 g of II-02, 168 mL of acetonitrile, and 42 mL of tap water were added to a flask; a solution of 38.739 g of EDCI, 30.948 g of HOBT, 108 mL of acetonitrile, and 32 mL of tap water was added, and the reaction was monitored by TLC or HPLC until complete; isopropyl acetate (600 mL) and tap water (420 mL) were added to the system, and the mixture was stirred for 10 min, separating the layers, and the organic phase was retained. The organic phase was washed twice with a solution of 11.3 g of sodium bicarbonate in 120 mL of water; the organic phase was evaporated to dryness, and acetone (100 mL) and water (840 mL) were added and the mixture was stirred for 1 h. The mixture was filtered and dried to obtain 61.9 g of white powder, which was intermediate III-02, with a molar yield of 91.6% and a purity of 98.725%.

[0066] (3) Intermediate IV-02: (2S,5R)-2-[2-((R)-1-(tert-butyloxycarbonyl)piperidin-3-carboxyl)hydrazine-1-carboxyl]-7-oxo-1,6-diazabicyclo[3.2.1]oct-6-yl sulfate tetrabutylammonium salt

[0067] At room temperature, III-02 (60.00 g), trimethylamine sulfur trioxide (23.806 g), 10% Pd / C (1.5 g), triethylamine (3.096 mL), isopropanol (240 mL), and water (240 mL) were added to a flask; the mixture was purged with nitrogen three times and hydrogen three times, and the reaction was detected to be complete by TLC or HPLC; palladium on carbon was removed by filtration, and the filtrate was washed once with isopropyl acetate (300 mL), retaining the aqueous phase; sodium bicarbonate (15.476 g) and tetrabutylammonium bisulfate (45.489 g) were added to the aqueous phase, followed by extraction with dichloromethane (420 mL), retaining the dichloromethane phase, which was then evaporated to dryness to obtain a pale yellow oil, which was intermediate IV-02 with a purity >98%. Based on a 100% yield, it was directly used for the next reaction.

[0068] (4) Zidebactam

[0069] At -10±5℃, the pale yellow oily substance IV-02 (approximately 87.68 g) and dichloromethane (87 mL) from the previous step were added to a flask. Under nitrogen protection, trifluoroacetic acid (210.68 mL) was added dropwise. The reaction was monitored by TLC or HPLC until complete. The system was then transferred to room temperature, and methyl tert-butyl ether (1700 mL) was added, resulting in the precipitation of a white solid. The solid was filtered to obtain crude zidabactam. The crude zidabactam was suspended in ethanol (600 mL) and water (87 mL), and sodium isooctanoate was added to adjust the pH to 6-7. Then, ethanol (450 mL) was added, and the mixture was stirred for 1 h. The mixture was filtered and dried to obtain 39.5 g of white powder, which was zidabactam, with a molar yield of 84.4% and a purity of 99.932%.

[0070] Example 3:

[0071]

[0072] (1) Intermediate II-03: (2-aminooxyethyl)carbamate tert-butyl ester-(2S,5R)-6-(benzyloxy)-7-oxo-1,6-diazabicyclo[3.2.1]octane-2-carboxylate

[0073] At room temperature, (2S,5R)-6-(benzyloxy)-7-oxo-1,6-diazabicyclo[3.2.1]octane-2-carboxylic acid benzyl ester I-03 (50.00 g) and tetrahydrofuran (250 mL) were added to a flask. The flask was then heated to -15°C externally and -10±5°C internally. A solution of lithium hydroxide (6.187 g) in water (400 mL) was added dropwise. After the addition was complete, the reaction was detected by TLC or HPLC to confirm its completeness. The flask was then cooled to room temperature, and dichloromethane (400 mL) was added. The pH was adjusted to 1-3 with 1 M hydrochloric acid. The mixture was allowed to separate into layers, and the organic phase was retained. A solution of β-lactamase inhibitor side chain (2-aminooxyethyl)carbamate tert-butyl ester (30.348 g) in dichloromethane (100 mL) was added. The mixture was stirred at room temperature for 1 h, filtered, and dried to obtain 73.6 g of the product. g of white powder, which is intermediate II-03, with a molar yield of 94.5% and a purity of 98.982%.

[0074] (2) Intermediate III-03: (2S,5R)-6-(benzyloxy)-7-oxo-1,6-diazabicyclo[3.2.1]octane-2-carboxylic acid[2-((tert-butoxycarbonyl)amino)ethoxy]amide

[0075] At room temperature, II-03 (70.00 g), acetonitrile (280 mL), and N,N-diisopropylethylamine (60 mL) were added to a flask; a BOP (82.152 g) solution in acetonitrile (210 mL) was added, and the reaction was monitored by TLC or HPLC until complete; dichloromethane (600 mL) and tap water (700 mL) were added to the system, stirred, and the mixture was allowed to separate into layers, retaining the organic phase. The organic phase was washed twice with a sodium bicarbonate (13.0 g) solution in water (120 mL); the organic phase was evaporated to dryness, and acetone (160 mL) and water (900 mL) were added and stirred for 1 h. The mixture was filtered and dried to obtain 63.6 g of white powder, which was intermediate III-03, with a molar yield of 94.6% and a purity of 98.225%.

[0076] (3) Intermediate IV-03:

[0077] At room temperature, III-03 (60.00 g), trimethylamine sulfur trioxide (27.482 g), 10% Pd / C (2.0 g), triethylamine (3.574 mL), isopropanol (240 mL), and water (240 mL) were added to a flask; the mixture was purged with nitrogen three times and hydrogen three times, and the reaction was detected to be complete by TLC or HPLC; the mixture was filtered to remove palladium on carbon, and the filtrate was washed once with isopropyl acetate (300 mL), retaining the aqueous phase; sodium bicarbonate (17.886 g) and tetrabutylammonium bisulfate (52.513 g) were added to the aqueous phase, followed by extraction with dichloromethane (420 mL), retaining the dichloromethane phase, which was then evaporated to dryness to obtain a pale yellow oil, which was intermediate IV-03 with a purity >98%. Based on a 100% yield, the reaction was directly carried out in the next step.

[0078] (4) Nacubactam

[0079] At -5±5℃, the pale yellow oily substance IV-03 (approximately 91.95 g) and dichloromethane (50 mL) from the previous step were added to a flask. Under nitrogen protection, trifluoroacetic acid (243.21 mL) was added dropwise. The reaction was monitored by TLC or HPLC until complete. The system was then transferred to room temperature, and methyl tert-butyl ether (1600 mL) was added, resulting in the precipitation of a white solid. The solid was filtered to obtain crude white nakubatane. The crude white nakubatane was suspended in acetonitrile (400 mL) and water (90 mL), and saturated sodium carbonate was added to adjust the pH to 6-7. Isopropanol (500 mL) was then added, and the mixture was stirred for 1 h. The mixture was filtered and dried to obtain 36.9 g of white powder, which was nakubatane, with a molar yield of 82.4% and a purity of 99.901%.

[0080] Example 4:

[0081] The preparation of β-lactamase inhibitor intermediate cocrystals or new salt compounds II-04, II-05, II-06, II-07, II-08, and II-09 can be carried out by referring to Examples 1, 2, and 3. The corresponding β-lactamase inhibitors can be easily prepared by referring to Examples 1, 2, and 3 using β-lactamase inhibitor intermediate cocrystals or new salt compounds.

Claims

1. A β-lactamase inhibitor intermediate salt, characterized in that, The structure is as follows: ； R2 represents either benzyl or allyl; R3 is selected from One of them; P represents one of the protecting groups: tert-butoxycarbonyl (Boc), benzyloxycarbonyl (Cbz), methoxycarbonyl (Fmoc), or allyloxycarbonyl (Alloc).

2. A β-lactamase inhibitor intermediate salt according to claim 1, characterized in that, Choose one of the following: 。 3. The method for preparing an intermediate salt of a β-lactamase inhibitor according to claim 1 or 2, characterized in that, The specific synthesis process route is shown below: ； The synthesis technology solution is as follows: Using compound I, compound I was dissolved in solvent A, and an alkaline solution was added to react under conditions below 0°C. Then, acid was added to adjust the pH to acidic, and solvent B was added to extract the product. The product was separated into layers, and the organic phase was retained. Then, the side chain R3H of the β-lactamase inhibitor was added to react. After the reaction was completed, a white solid precipitated. The solid was filtered to obtain the intermediate salt II of the β-lactamase inhibitor. In compound I, R1 represents an alkoxy group, selected from methoxy, ethoxy, propoxy, butoxy, or benzyloxy.

4. The method according to claim 3, characterized in that, Compound I is selected from one of the following: (2S,5R)-6-(benzyloxy)-7-oxo-1,6-diazabicyclo[3.2.1]octane-2-carboxylic acid ethyl ester, (2S,5R)-6-(benzyloxy)-7-oxo-1,6-diazabicyclo[3.2.1]octane-2-carboxylic acid methyl ester, or (2S,5R)-6-(benzyloxy)-7-oxo-1,6-diazabicyclo[3.2.1]octane-2-carboxylic acid benzyl ester; R3H is selected from One of them, P represents one of the protecting groups: tert-butoxycarbonyl (Boc), benzyloxycarbonyl (Cbz), methoxycarbonyl (Fmoc), or allyloxycarbonyl (Alloc).

5. The method according to claim 3, characterized in that, In the above steps, the molar ratio of compound I to the β-lactamase inhibitor side chain R3H is 1:0.8~1.

5.

6. The method according to claim 3, characterized in that, Solvent A is one or more of acetone, methanol, ethanol, isopropanol, tetrahydrofuran, and acetonitrile; the addition of acid and / or base includes water; solvent B is one or more of dichloromethane, chloroform, ethyl acetate, butyl acetate, isopropyl acetate, 1,2-dichloroethane, toluene, 2-methyltetrahydrofuran, cyclopentyl methyl ether, and chlorobenzene; base is selected from sodium hydroxide, lithium hydroxide, or potassium hydroxide; acid is selected from hydrochloric acid, sulfuric acid, nitric acid, acetic acid, and phosphoric acid.

7. The method according to claim 3, characterized in that, The temperature ranges from -20°C to -5°C when the temperature is below 0°C.

8. The use of the intermediate salt of the β-lactamase inhibitor according to claim 1 or 2 for the preparation of β-lactamase inhibitors.

9. A method for preparing β-lactamase inhibitors from an intermediate salt of a β-lactamase inhibitor as described in claim 1 or 2, characterized in that, Includes the following steps: ； Specifically, the steps include the following: (1) Dissolve the intermediate salt II of β-lactamase inhibitor in solvent C, add condensing agent, and after the reaction is complete, add solvent B' for extraction, separate the layers, retain the organic phase, wash with inorganic alkaline aqueous solution, rotary evaporate, crystallize, and obtain compound III. (2) Compound III is prepared by de-R2 reaction and sulfonation reaction in solvent D under catalytic conditions, followed by ammonium salt formation reaction; (3) In solvent E or without solvent, the compound of formula IV is deprotected by the protecting group P in R3 and then crystallized by adding a crystallization solvent to prepare β-lactamase inhibitor.

10. The method according to claim 9, characterized in that, In step (1), solvent C is an organic solvent or an organic solvent and water, and the organic solvent is selected from one or more of acetone, methanol, ethanol, isopropanol, tetrahydrofuran, and acetonitrile; solvent B' is an organic solvent or an organic solvent and water, and the organic solvent is selected from one or more of dichloromethane, chloroform, ethyl acetate, butyl acetate, isopropyl acetate, 1,2-dichloroethane, toluene, 2-methyltetrahydrofuran, cyclopentyl methyl ether, and chlorobenzene; at least one of solvent C and solvent B' contains water; in step (2), solvent D is an organic solvent or an organic solvent and water, and the organic solvent is selected from one or more of acetone, methanol, ethanol, isopropanol, tetrahydrofuran, acetonitrile, dichloromethane, chloroform, and ethyl acetate, and also includes organic bases; in step (3), solvent E is dichloromethane, chloroform, ethyl acetate, 2-methyltetrahydrofuran, and methyl tert-butyl ether; The condensing agents are 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI), 1-hydroxybenzotriazole (HOBt), N,N'-dicyclohexylcarbodiimide (DCC), N,N'-diisopropylcarbodiimide (DIC), 1-hydroxy-7-azabenzotriazole (HOAt), benzotriazole-N,N,N',N'-tetramethylurea hexafluorophosphate (HBTU), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (HATU), benzotriazole-1-yl-oxy-tris(dimethylamino)phosphohexafluorophosphate (BOP), and (benzotriazole) -1-yloxy)tripyrrolidinylphosphine hexafluorophosphate (PyBOP), N,N'-carbonyldiimidazole (CDI), propylphosphoric anhydride (T3P), triethylamine (TEA), N,N-diisopropylethylamine (DIPEA), pyridine, 4-dimethylaminopyridine (DMAP) or 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) or one or more thereof; in step (2), when R2 is benzyl or allyl, the removal of R2 is performed using hydrogen and palladium on carbon; sulfonation is selected from trimethylamine trioxide, triethylamine trioxide or pyridine trioxide complex; the ammonium salt formation reaction is selected from tetrabutylammonium acetate or tetrabutylammonium hydrogen sulfate; In step (3), under nitrogen protection, when the protecting group P is Boc, the de-Boc reagent is selected from trifluoroacetic acid; when the protecting group P is benzyloxycarbonyl (Cbz), the de-benzyloxycarbonyl (Cbz) reagent is selected from hydrogen bromide acetic acid solution; when the protecting group P is methoxycarbonyl (Fmoc), the de-methoxycarbonyl (Fmoc) reagent is selected from 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU); when the protecting group P is allyloxycarbonyl (Alloc), the de-allyloxycarbonyl (Alloc) reagent is tetra(triphenylphosphine)palladium; The crystallization solvent is selected from methyl tert-butyl ether.

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