CN107151254A - It is a kind of to be used as boric acid compound of 20S proteasome inhibitors and preparation method thereof - Google Patents

Abstract

The present invention relates to pharmaceutical chemistry and pharmacotherapeutics field.Specifically related to new boric acid compound and its production and use, it is especially a kind of to be used as boric acid compound of 20S proteasome inhibitors and preparation method thereof.The invention discloses boric acid compound of Formulas I and preparation method thereof, test, as a result show through bioactivity screening, obtained compound has the function of protease inhibition body, be further useful for preparing treatment and the medicine of proteasome relevant disease.

Description

A kind of boronic acid compound as 20S proteasome inhibitor and preparation method thereof

technical field

The present invention relates to the fields of medicinal chemistry and pharmacotherapeutics. It specifically relates to a new boric acid compound and its preparation method and application, especially a boric acid compound as a 20S proteasome inhibitor and its preparation method.

Background technique

Studies have reported that the Ubiquitin-Proteasome Pathway (UPP) is the main pathway for protein degradation in eukaryotic cells, which can regulate the level of proteins involved in cell cycle control and maintain the biological homeostasis of different cellular processes. Studies have shown that in most mammalian cells, this pathway can recognize and degrade misfolded proteins; a large number of experiments have proved that the pathogenesis of cancer, cardiovascular and cerebrovascular diseases, and neurodegenerative diseases are all related to the ubiquitin-proteasome pathway. connect.

Studies have disclosed that in the ubiquitin-proteasome pathway, the multi-catalyzed protease complex 26S proteasome is the main component of hydrolyzed proteins, composed of two regulatory particles 19S proteasome and a cylindrical 20S proteasome, 19S proteasome Located at both ends of the 20S proteasome, it consists of 18 subunits, which control the recognition, folding and translocation of protein substrates into the cavity of the 20S proteasome; the 20S proteasome consists of 28 subunits to form 4 stacked rings, of which 7 Three distinct α subunits (α1-α7) form the two outer compartments, and seven distinct β subunits (β1-β7) form the central compartment with the proteolytic active site; three of the relevant central compartments The proteolytic activity of subunits has been determined, including: β1 subunit has glutamyl transpeptide hydrolysis activity (PGPH), β2 subunit has trypsin-like activity (T-L), β5 subunit has chymotrypsin-like activity (CT -L).

Bortezomib is a covalent reversible selective proteasome inhibitor, which mainly inhibits chymotrypsin-like activity. It was launched in the United States by Millennium Pharmaceutical Company in 2003 and has been used clinically to treat multiple myeloma. However, clinical data show that bortezomib still has many side effects, such as diarrhea, vomiting, painful peripheral neuropathy, and thrombocytopenia. To date, the mechanism of these toxic side effects is unclear, and in addition, the clinical use of bortezomib requires a twice-weekly intravenous or subcutaneous injection regimen, which brings great inconvenience to patients. Therefore, it is of great practical significance to develop a novel proteasome inhibitor with low toxic and side effects, flexible administration and oral administration.

Contents of the invention

The purpose of the present invention is to overcome the defects of the prior art and provide a boric acid compound with novel structure and the function of inhibiting proteasome.

Another object of the present invention is to provide a preparation method of the above boric acid compound.

A further object of the present invention is to provide the use of the above-mentioned boronic acid compounds in the preparation of drugs for treating proteasome-related diseases.

specific,

The present invention provides the boronic acid compound of formula I structure,

in:

X is O or NH;

R is aryl or heteroaryl;

n is 0 or 1.

In some embodiments of the invention, the compound of formula I has the following structure:

in:

X is O or NH;

R is aryl or heteroaryl;

n is 0 or 1.

In some embodiments of the invention, the compound of formula I has the following structure:

in:

X is O or NH;

R is aryl or heteroaryl;

n is 0 or 1.

In some embodiments of the invention, the compound of formula I-a has the following structure:

in:

X is O or NH;

R is aryl or heteroaryl;

n is 0 or 1.

In some embodiments of the invention, the compound of formula I-a has the following structure:

in:

X is O or NH;

R is aryl or heteroaryl;

n is 0 or 1.

In some embodiments of the invention, the compound of formula I-b has the following structure:

in:

X is O or NH;

R is aryl or heteroaryl;

n is 0 or 1.

In some embodiments of the invention, the compound of formula I-b has the following structure:

in:

X is O or NH;

R is aryl or heteroaryl;

n is 0 or 1.

In the present invention,

The term "aryl" is used to indicate an aromatic carbocyclic group, including monocyclic or polycyclic aromatic hydrocarbons;

The term "heteroaryl" is used to indicate a substituted or unsubstituted aromatic carbocyclic group whose ring structure includes 1 to 4 heteroatoms; it is also used to indicate an aromatic carbocyclic group with two or more rings;

Dotted and bold lines are used to indicate the chemical structure of one or more stereocenters, indicating the absolute stereochemistry of the stereocenter in the chemical structure;

In the present invention, the bonds described in the structural diagrams do not represent a better stereostructure, and chemical structures containing one or more stereocenters include all possible stereoisomer forms and mixtures thereof;

The term "inhibitor" is used to denote a compound capable of blocking or reducing the activity of an enzyme, enzyme system, receptor or other pharmacological target.

The term "treating" is intended to include reversing, alleviating or arresting clinical conditions and associated pathological states, in order to ameliorate or stabilize a patient's condition.

The present invention further provides the preparation method of described compound:

The compounds of the present invention can be prepared by a variety of synthetic methods. Some of the raw materials used for the preparation of the compounds of the present invention can be obtained from commercial reagent companies, and the other part can be prepared by simple conversion methods known in the art.

Specifically, the compound of the general formula I shown can be prepared according to the following general synthetic route, wherein the substituents of the compound of the general formula I have the aforementioned meanings. This synthetic method is only illustrative to illustrate the present invention, not limiting it to the present invention. In the present invention, compound formula I-a is synthesized according to the following route:

Formula III-1 is obtained under the action of n-BuLi to obtain formula III-2, formula III-2 is reacted with B(OCH ₃ ) ₃ to obtain formula III-3, and formula III-3 and III-4-a are subjected to transesterification to obtain formula III-5-a, formula III-5-a is reacted with metal reagent i-BuMgBr, and then catalyzed by anhydrous ZnCl ₂ to obtain formula III-6-a, formula III-6-a is reacted with LiN(SiMe ₃ ) ₂ to obtain Formula III-7-a, formula III-7-a is deprotected under the action of HCl to obtain formula III-8-a, formula III-8-a and formula IV-1 are condensed to obtain formula IV-2-a, formula IV-2-a removes the group connected with B to obtain formula Ia;

The compound of formula I-b is synthesized according to the following route:

Formula III-1 is obtained under the action of n-BuLi to obtain formula III-2, formula III-2 is reacted with B(OCH ₃ ) ₃ to obtain formula III-3, and formula III-3 and III-4-b are reacted to obtain formula III-5-b, formula III-5-b is reacted with metal reagent i-BuMgBr, and then catalyzed by anhydrous ZnCl ₂ to obtain formula III-6-b, formula III-6-b is reacted with LiN(SiMe ₃ ) ₂ to obtain Formula III-7-b, formula III-7-b is deprotected under the action of HCl to obtain formula III-8-b, formula III-8-b and formula IV-1 are condensed to obtain formula IV-2-b, formula IV-2-b removes the group connected with B to obtain formula Ib.

More specifically, the present invention synthesizes the compound of general formula I according to the following route:

Wherein, the substituents of the compounds of general formula I have the meanings described above.

Synthetic formula I-a compound:

1) The compound of formula III-1 is reacted with n-BuLi at -110°C to obtain the organolithium intermediate shown in formula III-2; 2) The compound of formula III-2 is reacted with B at -110°C (OCH ₃ ) ₃ react to obtain the dimethyl borate structure shown in III-3; 3) the compound of the formula III-3 structure and the chiral pinanediol shown in the formula III-4-a are esterified at room temperature The exchange reaction obtains the borate ester structure shown in III-5-a;

4) The compound with the structure of formula III-5-a is first reacted with the metal reagent i- _BuMgBr at -78°C, and then catalyzed by anhydrous ZnCl2 at room temperature to obtain the borate ester shown in III-6-a structure;

5) The compound of the formula III-6-a is reacted with LiN(SiMe ₃ ) ₂ at -78°C to obtain the bis(trimethylsilyl)-protected aminoboronate structure shown in the formula III-7-a;

6) The compound of the formula III-7-a reacts with HCl at -78°C to deprotect the bis(trimethylsilyl) to obtain the unprotected amino borate hydrochloride shown in the formula III-8-a structure;

7) the compound of the formula III-8-a structure and the compound of the formula IV-1 structure in the condensation agent 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (abbreviated as EDC. HCl) and 1-hydroxybenzotriazole (abbreviated as HOBt) are reacted to obtain the borate structure shown in formula IV-2-a;

8) The compound of formula IV-2-a undergoes a transesterification reaction with 2-methylpropylboronic acid at room temperature to obtain the target boronic acid compound shown in formula I-a.

Synthetic formula I-b compound:

1) The compound of the formula III-1 is reacted with n-BuLi at -110°C to obtain the organolithium intermediate shown in the formula III-2;

2) The compound of formula III-2 is reacted with B(OCH ₃ ) ₃ at -110°C to obtain the dimethyl borate structure shown in III-3;

3) The compound of the formula III-3 and the chiral pinanediol shown in the formula III-4-b perform transesterification reaction at room temperature to obtain the boronate structure shown in III-5-b;

4) The compound of the formula III-5-b is first reacted with the metal reagent i- _BuMgBr at -78°C, and then catalyzed by anhydrous ZnCl at room temperature to obtain the borate ester shown in III-6-b structure;

5) The compound of the formula III-6-b is reacted with LiN(SiMe ₃ ) ₂ at -78°C to obtain the bis(trimethylsilyl)-protected aminoboronate structure shown in the formula III-7-b;

6. The compound of the formula III-7-b reacts with HCl at -78°C to deprotect the bis(trimethylsilyl) to obtain the unprotected amino borate hydrochloride shown in the formula III-8-b structure;

7) the compound of the formula III-8-b structure and the compound of the formula IV-1 structure in the condensation agent 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (abbreviated as EDC. HCl) and 1-hydroxybenzotriazole (abbreviated as HOBt) in the presence of the reaction to obtain the borate structure shown in formula IV-2-b;

8) The compound of the formula IV-2-b undergoes a transesterification reaction with 2-methylpropylboronic acid at room temperature to obtain the target boric acid compound shown in the formula I-b.

The present invention has carried out pharmacological research experiments, and the results show that the prepared compound of general formula I has good proteasome inhibitory activity, and some compounds show good proteasome inhibitory activity at the nanomolar level, and can be used to prepare proteasome inhibitory agent for the treatment of proteasome-related diseases.

detailed description

The present invention will be further described below in conjunction with examples, but these examples do not limit the scope of the present invention.

A part of the raw material used to prepare the compound of the present invention in the experimental method of the present invention can be derived from a commercial reagent company, and another part can be prepared using a concise conversion method well known in the art; ) determined; NMR was determined using a Varian400MHz nuclear magnetic resonance instrument, the measuring reagents were CDCl ₃ , CD ₃ OD and DMSO-d ₆ , the internal standard was TMS, and the chemical shift (δ) was in ppm; the LCMS was determined using Agilent Technologies6120 liquid mass spectrometry Instrument; column chromatography using YAMAZENAI-580S medium pressure preparative chromatography for product purification.

Embodiment 1: Preparation of dichloromethylenelithium (compound III-2)

_LiCHCl2

(III-2)

Under the condition of nitrogen protection, add anhydrous dichloromethane (4.6mL, 72mmol) to 200mL of anhydrous tetrahydrofuran, the temperature dropped to -110 ℃, dropwise added 1.6M n-butyllithium n-hexane solution (38mL, 60mmol ), after the dropwise addition, continued to stir at -110°C for 1 hour; the reaction solution was directly used in the next reaction without purification.

Embodiment 2: Preparation of dimethyl dichloromethylene borate (compound III-3)

Under the condition of nitrogen protection, the temperature was continued to be controlled at -110°C, trimethyl borate (8mL, 72mmol) was added to the compound III-2 solution prepared in Example 1, and stirring was continued at -110°C for 1 hour, and then Add 12mL of 5N HCl solution, the reaction is slowly raised to room temperature, transfer the reaction solution into a separatory funnel, separate the organic phase, extract the aqueous phase with ether (3×10mL), combine the organic phases, add anhydrous sodium sulfate to dry, dry After the completion of the filtration, the solvent was removed by a rotary evaporator to obtain 8.7 g of a white solid with a yield of 92%. The reaction product was directly used in the next reaction without purification.

Embodiment 3: Preparation (+)-α-pinanediol (compound III-4-a)

Under the condition of nitrogen protection, an aqueous solution of Me ₃ NO·2H ₂ O (11 g, 102 mmol) was added to 100 mL of tert-butanol, and (+)-α-pinene (15 mL, 97 mmol) and pyridine (7 mL) were sequentially added under stirring. and osmium tetroxide (51mg, 0.2mmol), then warming up to reflux, TLC detection after 24 hours showed that the reaction was complete, the reaction was down to room temperature, added NaHSO ₃ (1.2g, 12mmol) stirred for 1 hour, the reaction solution was transferred to the separation In the funnel, the organic phase was separated, the aqueous phase was extracted with ether (3×20 mL), the organic phases were combined, dried by adding anhydrous sodium sulfate, filtered after drying, the solvent was removed by a rotary evaporator, and separated by column chromatography (petroleum ether: Ethyl acetate=30:1) 15 g of white solid was obtained, and the yield was 91%.

Example 4: Preparation of Dichloromethyleneboronic Acid-(+)-α-Pinanediol Ester (Compound III-5-a)

Add Example 2 Compound III-3 (10g, 63mmol) and Example 3 Compound III-4-a (7.2g, 42mmol) to 150mL of anhydrous tetrahydrofuran, stir at room temperature, TLC detection after 18 hours shows that the reaction is complete. The solvent was removed by an evaporator and separated by column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain 10 g of a colorless oily liquid with a yield of 91%. ¹ H NMR(CDCl ₃ ,400MHz)δ5.39(s,1H),4.46(d,1H,J=8.8Hz),2.33(m,2H),2.12(t,1H,J=5.2Hz),1.94 (m, 2H), 1.46(s, 3H), 1.30(s, 3H), 1.21(d, 1H, J=11.2Hz), 0.84(s, 3H).

Example 5: Preparation of 2-methyl-4-chloro-butylboronic acid-(+)-α-pinanediol ester (compound III-6-a)

Under the condition of nitrogen protection, compound III-5 (10 g, 38 mmol) of Example 4 was added to 100 mL of anhydrous ether, the temperature was lowered to -78 ° C, and a 2M ether solution of isobutylmagnesium bromide (17 mL, _33mmol ), after the dropwise addition, dry ZnCl powder was added, and the reaction was slowly raised to room temperature. After 20 hours, TLC detection showed that the reaction was complete, and the solid was removed by filtration, and the rotary evaporator was used to remove the solvent, and the column chromatography was separated (petroleum ether: Ethyl acetate=200:1) to obtain 7.6 g of colorless oily liquid, yield 81%. ¹ H NMR(CDCl ₃ ,400MHz)δ4.36(d,1H,J=9.1Hz),3.52(m,1H),2.35(m,1H),2.24(m,1H),2.08(t,1H, J＝5.4Hz), 1.92(m, 2H), 1.87(d, 1H, J＝1.6Hz), 1.79(m, 1H), 1.62(m, 1H), 1.41(s, 3H), 1.29(s, 3H), 1.18(d, 1H, J=11.0Hz), 0.92(d, 3H, J=6.6Hz), 0.90(d, 3H, J=6.6Hz), 0.84(s, 3H).

Example 6: Preparation of 2-methyl-4-amino-butylboronic acid-(+)-α-pinanediol ester hydrochloride (compound III-8-a)

Under nitrogen protection, add Example 5 compound III-6-a (4.5 g, 16 mmol) to 100 mL of anhydrous tetrahydrofuran, drop the temperature to -78 ° C, and add 1M LiN(SiMe ₃ ) ₂ tetrahydrofuran solution dropwise (24mL, 24mmol), after the dropwise addition, the reaction was slowly raised to room temperature, and after 24 hours, TLC detection showed that the reaction was complete, and the solvent was removed by a rotary evaporator, 50mL of n-hexane was added to dissolve, the solid was removed by filtration, and the temperature of the filtrate was reduced to -78 ℃, 2M HCl ether solution (24mL, 48mmol) was added, the reaction was slowly raised to room temperature, and a large amount of white solid appeared, which was filtered and washed with ether to obtain 3.2g of white solid, with a yield of 66%. ¹ H NMR(DMSO-d6,400MHz)δ7.84(s,3H),7.41(s,1H),7.29(s,1H),7.16(s,1H),4.44(m,1H),2.74(m ,1H),2.32(m,1H),2.18(m,1H),1.99(t,1H,J=5.5Hz),1.88(m,1H),1.73(m,2H),1.47(m,2H) , 1.35(s, 3H), 1.24(s, 3H), 1.11(d, 1H, J=10.8Hz), 0.86(d, 6H, J=6.5Hz), 0.81(s, 3H).

Embodiment 7: Preparation of (-)-α-pinanediol (compound III-4-b)

Under nitrogen protection conditions, add Me ₃ NO·2H ₂ O (11g, 102mmol) aqueous solution to 100mL tert-butanol, add (-)-α-pinene (15mL, 97mmol) and pyridine (7mL) sequentially under stirring and osmium tetroxide (51mg, 0.2mmol), then warming up to reflux, after 20 hours, TLC detection showed that the reaction was complete, and the reaction was down to room temperature, adding NaHSO ₃ (1.2g, 12mmol) was stirred for 1 hour, and the reaction solution was transferred to the separatory In the funnel, the organic phase was separated, the aqueous phase was extracted with ether (3×20 mL), the organic phases were combined, dried by adding anhydrous sodium sulfate, filtered after drying, the solvent was removed by a rotary evaporator, and separated by column chromatography (petroleum ether: Ethyl acetate=30:1) 14 g of white solid was obtained, and the yield was 85%.

Example 8: Preparation of Dichloromethyleneboronic Acid-(-)-α-Pinanediol Ester (Compound III-5-b)

Add Example 2 Compound III-3 (10g, 63mmol) and Example 7 Compound III-4-b (7.2g, 42mmol) to 150mL of anhydrous tetrahydrofuran, stir at room temperature, TLC detection after 23 hours shows that the reaction is complete. The solvent was removed by an evaporator and separated by column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain 9 g of a colorless oily liquid with a yield of 81%. ¹ H NMR(CDCl ₃ ,400MHz)δ5.39(s,1H),4.46(d,1H,J=8.7Hz),2.33(m,2H),2.12(t,1H,J=5.4Hz),1.94 (m, 2H), 1.46(s, 3H), 1.30(s, 3H), 1.20(d, 1H, J=11.2Hz), 0.84(s, 3H).

Example 9: Preparation of 2-methyl-4-chloro-butylboronic acid-(-)-α-pinanediol ester (compound III-6-b)

Under the condition of nitrogen protection, the compound III-5-b (10 g, 38 mmol) of Example 8 was added to 100 mL of anhydrous ether, the temperature was lowered to -78 ° C, and a 2M isobutylmagnesium bromide ether solution was added dropwise ( 17mL, _33mmol ), after the dropwise addition, add dry ZnCl powder, the reaction is slowly raised to room temperature, after 22 hours, TLC detection shows that the reaction is complete, and the solid is removed by filtration, and the rotary evaporator is used to remove the solvent, and the column chromatography separates (petroleum Ether: ethyl acetate = 200:1) to obtain 5.8 g of a colorless oily liquid with a yield of 62%. ¹ H NMR(CDCl ₃ ,400MHz)δ4.36(d,1H,J=8.8Hz),3.52(m,1H),2.35(m,1H),2.24(m,1H),2.08(t,1H, J=5.4Hz), 1.91(s,2H), 1.87(s,1H), 1.79(m,1H), 1.61(m,1H), 1.41(s,3H), 1.29(s,3H), 1.18( d, 1H, J=11.1Hz), 0.92(d, 3H, J=6.6Hz), 0.90(d, 3H, J=6.6Hz), 0.84(s, 3H).

Example 10: Preparation of 2-methyl-4-amino-butylboronic acid-(-)-α-pinanediol ester hydrochloride (compound III-8-b)

Under nitrogen protection, add Example 9 compound III-6-b (4.5 g, 16 mmol) to 100 mL of anhydrous tetrahydrofuran, drop the temperature to -78 ° C, and add 1M LiN(SiMe ₃ ) ₂ tetrahydrofuran solution dropwise (24mL, 24mmol), after the dropwise addition, the reaction was slowly raised to room temperature, after 18 hours, TLC detection showed that the reaction was complete, and the solvent was removed by rotary evaporator, 50mL of n-hexane was added to dissolve, the solid was removed by filtration, and the temperature of the filtrate was reduced to -78 ℃, 2M HCl ether solution (24mL, 48mmol) was added, the reaction was slowly raised to room temperature, and a large amount of white solid appeared, which was filtered and washed with ether to obtain 4.1g of white solid, with a yield of 85%. ¹ H NMR(DMSO-d6,400MHz)δ7.85(s,3H),7.42(s,1H),7.29(s,1H),7.16(s,1H),4.45(d,1H,J＝8.3Hz ),2.76(m,1H),2.32(m,1H),2.20(m,1H),2.00(t,1H,J＝5.3Hz),1.88(s,1H),1.73(m,2H),1.48 (t,2H,J=7.3Hz),1.36(s,3H),1.25(s,3H),1.11(dd,1H,J=5.5Hz,J=10.6Hz),0.86(d,6H,J= 4.8Hz), 0.82(s,3H).

Example 11: Preparation of (S)-2-phenoxy-3-phenylpropionic acid methyl ester

Under nitrogen protection, (R)-2-hydroxyl-3-phenylpropanoic acid methyl ester (1.1g, 6mmol), phenol (0.38g, 4mmol) and triphenylphosphine ( 1.6g, 6mmol), the temperature was lowered to 0 ° C and stirred, and the anhydrous toluene solution of DIAD (1.2g, 6mmol) was added dropwise, and the reaction was slowly raised to room temperature. After 24 hours, TLC detection showed that the reaction was complete. Using a rotary evaporator Remove the solvent, add 10 mL of diethyl ether and 10 mL of n-hexane to the residue, remove the solid by filtration, use a rotary evaporator to remove the solvent, and separate by column chromatography (petroleum ether: ethyl acetate = 50:1) to obtain 0.58 g of the product, with a yield of 57 %. ¹ H NMR (CDCl ₃ , 400MHz) δ7.30(d, 4H, J=4.4Hz), 7.23(m, 3H), 6.95(t, 1H, J=7.4Hz), 6.83(d, 2H, J= 7.8Hz), 4.81(m,1H), 3.72(s,3H), 3.25(m,2H). MS(ESI) m/z257.0[M+H] ⁺ ,279.0[M+Na] ⁺ .

Preparation of (S)-2-phenoxy-3-phenylpropionic acid

Add (S)-2-phenoxy-3-phenylpropionic acid methyl ester (0.35g, 1.4mmol) to 5mL tetrahydrofuran, stir at room temperature, add 7mL of 1M sodium hydroxide solution, TLC detection shows reaction after 6 hours Completely, use a rotary evaporator to remove the solvent, add 6M HCl solution to the residue to acidify, use ether to extract (3×5mL), combine the organic phases, add anhydrous sodium sulfate to dry, filter after drying, and use a rotary evaporator to remove Solvent was used to obtain 0.33 g of white solid, with a yield of 97%. ¹ H NMR (CDCl ₃ , 400MHz) δ9.94(s, 1H), 7.29(m, 7H), 6.98(t, 1H, J=7.4Hz), 6.85(d, 2H, J=8.2Hz), 4.85 (m,1H),3.29(m,2H).MS(ESI)m/z241.0[MH] ^- .

Preparation of ((R)-3-methyl-1-((S)-2-phenoxy-3-phenylpropionamido)butyl)boronic acid (compound I-1)

Under nitrogen protection, (S)-2-phenoxy-3-phenylpropionic acid (27 mg, 0.11 mmol) was added to 5 mL of anhydrous dichloromethane, stirred at room temperature, HOBt (18 mg, 0.13 mmol) was added, After continuing to stir for 10 minutes, EDC·HCl (25mg, 0.13mmol) was added, followed by the addition of Example 6 compound III-8-a (50mg, 0.17mmol) and DIPEA (0.04mL, 0.22mmol), after 13 hours TLC detection showed a reaction Completely, the reaction solution was washed with 5% _NaHCO3 solution, 10% citric acid solution, 5% _NaHCO3 solution and saturated brine, and dried by adding anhydrous sodium sulfate to the organic phase, filtered after drying, and removed by rotary evaporator. solvent, the resulting product was directly added to 5 mL of methanol, stirred at room temperature, added 2-methylpropylboronic acid (22 mg, 0.22 mmol) and 5 mL of n-hexane, then added 1M HCl solution (0.33 mL, 0.33 mmol), and after 5 hours TLC detection shows that the reaction is complete, the reaction solution is allowed to stand until the layers are separated, the lower layer is washed with n-hexane, dried by adding anhydrous sodium sulfate, filtered after drying, the solvent is removed by a rotary evaporator, and column chromatography is separated (dichloromethane:methanol= 100:1) 17 mg of white solid was obtained, and the yield was 44%. ¹ H NMR (CD ₃ OD, 400MHz) δ7.26(m, 7H), 6.96(m, 3H), 5.11(m, 1H), 3.26(m, 2H), 2.66(t, 1H, J=7.3Hz ),1.48(m,1H),1.24(t,2H,J＝7.1Hz),0.81(d,6H,J＝6.6Hz).MS(ESI)m/z354.0[MH] ^- ,368.0[M +CH ₂ -H] ^- ,382.0[M+2CH ₂ -H] ^- .

Example 12: Preparation of (S)-2-(1-naphthyloxy)-3-phenylpropionic acid methyl ester

Under nitrogen protection, (R)-2-hydroxy-3-phenylpropanoic acid methyl ester (1.1g, 6mmol), 1-naphthol (0.58g, 4mmol) and triphenyl Phosphine (1.6g, 6mmol), the temperature was lowered to 0°C and stirred, and the anhydrous toluene solution of DIAD (1.2g, 6mmol) was added dropwise, and the reaction was slowly raised to room temperature. After 24 hours, TLC detection showed that the reaction was complete. Remove the solvent with a rotary evaporator, add 10 mL of diethyl ether and 10 mL of n-hexane to the residue, remove the solid by filtration, remove the solvent with a rotary evaporator, and separate by column chromatography (petroleum ether: ethyl acetate = 60:1) to obtain 0.56 g of the product. Yield 46%. ¹ H NMR(CDCl ₃ ,400MHz)δ8.31(m,1H),7.78(m,1H),7.40(m,8H),7.24(m,1H),6.64(d,1H,J＝7.6Hz) ,5.03(m,1H),3.71(s,3H),3.41(m,2H).MS(ESI) m/z307.0[M+H] ⁺ ,329.0[M+Na] ⁺ .

Preparation of (S)-2-(1-naphthyloxy)-3-phenylpropionic acid

Add (S)-2-(1-naphthyloxy)-3-phenylpropanoic acid methyl ester (0.56g, 1.8mmol) to 5mL tetrahydrofuran, stir at room temperature, add 9mL of 1M sodium hydroxide solution, after 6 hours TLC detection showed that the reaction was complete. Use a rotary evaporator to remove the solvent, add 6M HCl solution to acidify the residue, extract with ether (3×5mL), combine the organic phases, add anhydrous sodium sulfate to dry, filter after drying, and use The solvent was removed by a rotary evaporator to obtain 0.52 g of a white solid with a yield of 99%. ¹ H NMR(CDCl ₃ ,400MHz)δ9.68(s,1H),8.27(m,1H),7.79(m,1H),7.39(m,9H),6.67(d,1H,J＝7.7Hz) ,5.07(m,1H),3.44(m,2H).MS(ESI)m/z291.0[MH] ^- .

Preparation of ((R)-3-methyl-1-((S)-2-(1-naphthyloxy)-3-phenylpropionamido)butyl)boronic acid (compound 1-2)

Under nitrogen protection, (S)-2-(1-naphthyloxy)-3-phenylpropionic acid (32 mg, 0.11 mmol) was added to 5 mL of anhydrous dichloromethane, stirred at room temperature, and HOBt (18 mg, 0.13mmol), continue to stir for 10 minutes and add EDC·HCl (25mg, 0.13mmol), then add Example 6 compound III-8-a (50mg, 0.17mmol) and DIPEA (0.04mL, 0.22mmol), after 16 hours TLC detection showed that the reaction was complete, and the reaction solution was washed with ₅ % NaHCO solution, 10% citric acid solution, ₅ % NaHCO solution and saturated brine, and dried by adding anhydrous sodium sulfate to the organic phase, filtered after drying, and used The solvent was removed by a rotary evaporator, and the resulting product was directly added to 5 mL of methanol, stirred at room temperature, 2-methylpropylboronic acid (22 mg, 0.22 mmol) and 5 mL of n-hexane were added, and then 1M HCl solution (0.33 mL, 0.33 mmol) was added After 5 hours, TLC detection showed that the reaction was complete, and the reaction solution was allowed to stand until the layers were separated. The lower layer was washed with normal hexane, dried by adding anhydrous sodium sulfate, and filtered after drying, using a rotary evaporator to remove the solvent, and separated by column chromatography (dichloro Methane:methanol=80:1) to obtain 15 mg of white solid with a yield of 34%. ¹ H NMR (CD ₃ OD, 400MHz) δ8.24(m, 1H), 7.80(m, 1H), 7.36(m, 9H), 6.84(t, 1H, J=7.9Hz), 5.29(m, 1H ),3.42(m,2H),2.70(t,1H,J=7.1Hz),1.43(m,1H),1.19(t,2H,J=6.1Hz),0.78(d,6H,J=7.3Hz ).MS(ESI) m/z404.0[MH] ^- ,418.0[M+CH ₂ -H] ^- ,432.0[M+2CH ₂ -H] ^- .

Example 13: Preparation of (S)-3-phenyl-2-(2-pyrazinyloxy)propionic acid methyl ester

Under nitrogen protection, (R)-2-hydroxy-3-phenylpropanoic acid methyl ester (1.1 g, 6 mmol), 2-hydroxypyrazine (0.38 g, 4 mmol) and tris Phenylphosphine (1.6g, 6mmol), the temperature was lowered to 0°C and stirred, and anhydrous toluene solution of DIAD (1.2g, 6mmol) was added dropwise, and the reaction was slowly raised to room temperature. After 29 hours, TLC detection showed that the reaction was complete. Use a rotary evaporator to remove the solvent, add 10 mL of ether and 10 mL of n-hexane to the residue, filter to remove the solid, use a rotary evaporator to remove the solvent, and separate by column chromatography (petroleum ether: ethyl acetate = 30:1) to obtain 0.47 g of the product , yield 45%. ¹ HNMR (CDCl ₃ , 400MHz) δ8.30 (d, 1H, J = 1.1Hz), 8.12 (d, 1H, J = 2.8Hz), 7.98 (m, 1H), 7.30 (m, 5H), 5.42 ( m,1H),3.71(s,3H),3.28(m,2H).MS(ESI)m/z259.0[M+H] ⁺ ,281.0[M+Na] ⁺ .

Preparation of (S)-3-phenyl-2-(2-pyrazinyloxy)propionic acid

Add (S)-3-phenyl-2-(2-pyrazinyloxy)propionic acid methyl ester (0.47g, 1.8mmol) to 5mL tetrahydrofuran, stir at room temperature, add 9mL of 1M sodium hydroxide solution, 5 hours After TLC detection showed that the reaction was complete, the solvent was removed by a rotary evaporator, acidified by adding 6M HCl solution to the residue, extracted with ether (3×5mL), the organic phases were combined, dried by adding anhydrous sodium sulfate, and filtered after drying. The solvent was removed by a rotary evaporator to obtain 0.39 g of a white solid with a yield of 89%. ¹ H NMR(CDCl ₃ ,400MHz)δ8.29(s,1H),8.12(s,1H),8.06(s,1H),7.31(m,5H),5.46(m,1H),3.32(m, 2H).MS(ESI)m/z245.0[M+H] ⁺ ,266.9[M+Na] ⁺ .

Preparation of ((R)-3-methyl-1-((S)-3-phenyl-2-(2-pyrazinyloxy)propionamido)butyl)boronic acid (compound I-3)

Under nitrogen protection, (S)-3-phenyl-2-(2-pyrazinyloxy)propionic acid (27 mg, 0.11 mmol) was added to 5 mL of anhydrous dichloromethane, stirred at room temperature, and HOBt (18 mg , 0.13mmol), continued to stir for 10 minutes and added EDC·HCl (25mg, 0.13mmol), then added Example 6 compound III-8-a (50mg, 0.17mmol) and DIPEA (0.04mL, 0.22mmol), 17 hours Afterwards, TLC detection showed that the reaction was complete, and the reaction solution was washed with ₅ % NaHCO solution, 10% citric acid solution, ₅ % NaHCO solution and saturated brine, and dried by adding anhydrous sodium sulfate to the organic phase, and filtered after drying. Utilize the rotary evaporator to remove solvent, the obtained product is directly added in 5mL methanol, stir at room temperature, add 2-methylpropylboronic acid (22mg, 0.22mmol) and 5mL n-hexane, then add 1M HCl solution (0.33mL, 0.33mmol ), after 7 hours, TLC detection showed that the reaction was complete, and the reaction solution was allowed to stand until the layers were separated, and the lower layer was washed with normal hexane, dried by adding anhydrous sodium sulfate, and filtered after drying, using a rotary evaporator to remove the solvent, and separated by column chromatography (two Chloromethane:methanol=60:1) to obtain 18 mg of yellow solid with a yield of 46%. ¹ H NMR (CD ₃ OD, 400MHz) δ8.30(s, 1H), 8.17(t, 1H, J=2.6Hz), 8.10(m, 1H), 7.26(m, 5H), 5.72(m, 1H ), 3.34(d, 2H, J=4.9Hz), 2.65(t, 1H, J=7.3Hz), 1.51(m, 1H), 1.24(t, 2H, J=7.1Hz), 0.83(d, 6H ,J=6.7Hz).MS(ESI)m/z356.0[MH] ^- ,370.0[M+CH ₂ -H] ^- ,384.0[M+2CH ₂ -H] ^- .

Example 14: Preparation of (S)-3-phenyl-2-(phenylamino)propionic acid

Under nitrogen protection, (S)-2-amino-3-phenylpropionic acid (125 mg, 0.75 mmol), bromobenzene (80 mg, 0.5 mmol) were added to 10 mL of anhydrous N, N-dimethylacetamide in sequence ), potassium carbonate (140mg, 1mmol) and cuprous iodide (15mg, 0.08mmol), the temperature was raised to 90°C and stirred, after 48 hours, TLC detection showed that the reaction was complete, and the reaction was cooled to room temperature, and 5mL ethyl acetate and 1mL water were added , the temperature was lowered to 0°C, the pH was adjusted to 3 with 6M HCl solution, the organic phase was separated, the aqueous phase was extracted with ethyl acetate (3×10mL), the organic phases were combined, dried by adding anhydrous sodium sulfate, and filtered after drying , using a rotary evaporator to remove the solvent, and separated by column chromatography (petroleum ether: ethyl acetate = 2:1) to obtain 69 mg of the product, with a yield of 57%. ¹ H NMR (CDCl ₃ , 400MHz) δ7.31(m, 4H), 7.20(t, 3H, J=6.3Hz), 6.84(t, 1H, J=7.2Hz), 6.65(d, 2H, J= 5.2Hz),4.30(m,1H),3.21(m,2H).MS(ESI)m/z240.0[MH] ^- .

Preparation of ((R)-3-methyl-1-((S)-3-phenyl-2-(phenylamino)propionamido)butyl)boronic acid (compound I-4)

Under nitrogen protection, (S)-3-phenyl-2-(phenylamino)propionic acid (27 mg, 0.11 mmol) was added to 5 mL of anhydrous dichloromethane, stirred at room temperature, and HOBt (18 mg, 0.13 mmol) was added , add EDC·HCl (25mg, 0.13mmol) after continuing stirring for 10 minutes, then add Example 6 compound III-8-a (50mg, 0.17mmol) and DIPEA (0.04mL, 0.22mmol), after 8 hours TLC detection shows After the reaction is complete, the reaction solution is washed with 5% _NaHCO3 solution, 10% citric acid solution, 5% _NaHCO3 solution and saturated brine, and dried by adding anhydrous sodium sulfate to the organic phase. In addition to solvent, the resulting product was directly added to 5 mL of methanol, stirred at room temperature, 2-methylpropylboronic acid (22 mg, 0.22 mmol) and 5 mL of n-hexane were added, and then 1M HCl solution (0.33 mL, 0.33 mmol) was added for 5 hours After the TLC detection shows that the reaction is complete, the reaction solution is allowed to stand until the layers are separated, the lower layer is washed with n-hexane, dried by adding anhydrous sodium sulfate, filtered after drying, the solvent is removed by a rotary evaporator, and column chromatography is separated (dichloromethane: methanol =70:1) 14 mg of white solid was obtained, the yield was 36%. ¹ H NMR (CD ₃ OD, 400MHz) δ7.28(m, 7H), 7.00(m, 3H), 4.43(m, 1H), 3.22(m, 2H), 2.69(t, 1H, J＝6.6Hz ),1.24(m,1H),1.04(t,2H,J＝6.2Hz),0.73(d,6H,J＝7.2Hz).MS(ESI)m/z353.0[MH] ^- ,367.0[M +CH ₂ -H] ^- ,380.9[M+2CH ₂ -H] ^- .

Example 15: Preparation of (S)-2-(1-naphthylamino)-3-phenylpropionic acid

Under nitrogen protection, (S)-2-amino-3-phenylpropionic acid (1.2 g, 7.2 mmol), 1-bromonaphthalene (1 g , 4.8mmol), potassium carbonate (1.3g, 9.6mmol) and cuprous iodide (0.13g, 0.7mmol), the temperature was raised to 90°C and stirred, after 48 hours TLC detection showed that the reaction was complete, the reaction was cooled to room temperature, and 30mL Ethyl acetate and 10mL water, the temperature was lowered to 0°C, the pH was adjusted to 3 with 6M HCl solution, the organic phase was separated, the aqueous phase was extracted with ethyl acetate (3×20mL), the organic phase was combined, and anhydrous sodium sulfate was added Dry, filter after drying, remove the solvent with a rotary evaporator, and separate by column chromatography (petroleum ether: ethyl acetate = 2:1) to obtain 0.7 g of the product, with a yield of 50%. ¹ H NMR(DMSO-d6,400MHz)δ12.77(s,1H),8.25(m,1H),7.76(m,1H),7.29(m,8H),6.39(m,2H),4.26(d ,1H,J＝0.7Hz),3.26(m,2H).MS(ESI)m/z289.8[MH] ^- .

Preparation of ((R)-3-methyl-1-((S)-2-(1-naphthylamino)-3-phenylpropionamido)butyl)boronic acid (compound I-5)

Under nitrogen protection, (S)-2-(1-naphthylamino)-3-phenylpropanoic acid (32 mg, 0.11 mmol) was added to 5 mL of anhydrous dichloromethane, stirred at room temperature, and HOBt (18 mg, 0.13 mmol), continue to stir for 10 minutes and add EDC·HCl (25mg, 0.13mmol), then add Example 6 compound III-8-a (50mg, 0.17mmol) and DIPEA (0.04mL, 0.22mmol), TLC after 16 hours Detection showed that the reaction was complete, and the reaction solution was washed with 5% _NaHCO3 solution, 10% citric acid solution, 5% _NaHCO3 solution and saturated brine, and dried by adding anhydrous sodium sulfate to the organic phase. The evaporator was used to remove the solvent, and the resulting product was directly added to 5 mL of methanol, stirred at room temperature, 2-methylpropylboronic acid (22 mg, 0.22 mmol) and 5 mL of n-hexane were added, and then 1M HCl solution (0.33 mL, 0.33 mmol) was added. After 7 hours, TLC detection showed that the reaction was complete, and the reaction solution was allowed to stand until the layers were separated. The lower layer was washed with normal hexane, dried by adding anhydrous sodium sulfate, filtered after drying, and desolvated by a rotary evaporator, and separated by column chromatography (dichloromethane) :methanol=50:1) to obtain 12 mg of white solid, yield 27%. ¹ H NMR (CD ₃ OD, 400MHz) δ8.10(d, 1H, J=7.9Hz), 7.92(m, 1H), 7.76(d, 1H, J=8.1Hz), 7.62(m, 2H), 7.35(m,6H),7.12(d,1H,J=7.2Hz),4.69(m,1H),3.55(m,2H),2.80(t,1H,J=7.9Hz),1.26(m,1H ),0.98(t,2H,J=7.2Hz),0.66(d,6H,J=7.4Hz).MS(ESI)m/z403.3[MH] ^- ,417.3[M+CH ₂ -H] ^- ,431.3[M+2CH ₂ -H] ^- .

Example 16: Preparation of (S)-3-phenyl-2-(2-pyrazineamino)propionic acid

Under nitrogen protection, (S)-2-amino-3-phenylpropionic acid (1.6 g, 9.5 mmol), 2-bromopyrazine ( 1g, 6.3mmol), potassium carbonate (1.7g, 12.6mmol) and cuprous iodide (0.2g, 0.9mmol), the temperature was raised to 90°C and stirred, and TLC detection after 48 hours showed that the reaction was complete, and the reaction was cooled to room temperature, and added 30mL ethyl acetate and 10mL water, lower the temperature to 0°C, adjust the pH to 3 with 6M HCl solution, separate the organic phase, extract the aqueous phase with ethyl acetate (3×20mL), combine the organic phases, add anhydrous sulfuric acid Dry over sodium, filter after drying, remove the solvent with a rotary evaporator, and separate by column chromatography (petroleum ether: ethyl acetate = 1:1) to obtain 0.7 g of the product, with a yield of 46%. ¹ H NMR (DMSO-d6,400MHz)δ12.66(s,1H),7.99(d,1H,J=1.4Hz),7.87(m,1H),7.65(d,1H,J=2.8Hz), 7.40(d,1H,J=8.1Hz),7.26(d,3H,J=4.3Hz),7.18(m,1H),4.55(m,1H),3.14(m,1H),2.96(m,1H ).MS(ESI)m/z242.0[MH] ^- .

Preparation of ((R)-3-methyl-1-((S)-3-phenyl-2-(2-pyrazineamino)propionamido)butyl)boronic acid (compound I-6)

Under nitrogen protection, (S)-3-phenyl-2-(2-pyrazineamino)propionic acid (27 mg, 0.11 mmol) was added to 5 mL of anhydrous dichloromethane, stirred at room temperature, and HOBt (18 mg, 0.13mmol), continue to stir for 10 minutes and add EDC·HCl (25mg, 0.13mmol), then add Example 6 compound III-8-a (50mg, 0.17mmol) and DIPEA (0.04mL, 0.22mmol), after 13 hours TLC detection showed that the reaction was complete, and the reaction solution was washed with ₅ % NaHCO solution, 10% citric acid solution, ₅ % NaHCO solution and saturated brine, and dried by adding anhydrous sodium sulfate to the organic phase, filtered after drying, and used The solvent was removed by a rotary evaporator, and the resulting product was directly added to 5 mL of methanol, stirred at room temperature, 2-methylpropylboronic acid (22 mg, 0.22 mmol) and 5 mL of n-hexane were added, and then 1M HCl solution (0.33 mL, 0.33 mmol) was added After 6 hours, TLC detection showed that the reaction was complete, and the reaction solution was allowed to stand until the layers were separated. The lower layer was washed with normal hexane, dried by adding anhydrous sodium sulfate, and filtered after drying, using a rotary evaporator to remove the solvent, and separated by column chromatography (dichloro Methane:methanol=50:1) to obtain 12mg of yellow solid with a yield of 31%. ¹ H NMR (CD ₃ OD, 400MHz) δ7.99(s, 1H), 7.54(d, 2H, J=5.8Hz), 7.08(m, 5H), 3.98(m, 1H), 2.99(d, 2H ,J=55.0Hz), 2.19(t,1H,J=7.6Hz),1.73(m,1H),1.62(t,2H,J=5.7Hz),0.94(d,6H,J=6.1Hz). MS (ESI) m/z 369.1 [M+CH ₂ -H] ^- , 383.3 [M + 2CH ₂ -H] ^- .

Example 17: Preparation of (S)-2-(benzyloxy)-3-phenylpropionic acid methyl ester

Add (S)-2-hydroxy-3-phenylpropionic acid methyl ester (2.1g, 11.5mmol) to 50mL dichloromethane and 50mL n-hexane mixed solvent, stir at room temperature, dropwise add 2,2,2- Trichloroacetamide benzyl ester (3.5g, 13.8mmol), after dropwise addition, add the trifluoromethanesulfonic acid of catalytic amount, after 10 hours, TLC detects and shows that reaction is complete, utilizes rotary evaporator to remove solvent, and column chromatography separates ( Petroleum ether: ethyl acetate = 50:1) to obtain 1.3 g of the product with a yield of 42%. ¹ H NMR(CDCl ₃ ,400MHz)δ7.22(m,10H),4.67(d,1H,J=11.9Hz),4.37(d,1H,J=11.9Hz),4.14(m,1H),3.72 (s,3H),3.06(m,2H).MS(ESI)m/z271.2[M+H] ⁺ ,293.2[M+Na] ⁺ .

Preparation of (S)-2-(benzyloxy)-3-phenylpropionic acid

Add (S)-2-(benzyloxy)-3-phenylpropanoic acid methyl ester (1.3g, 4.8mmol) to 20mL of dioxane, stir at room temperature, add 24mL of 2M HCl solution, and raise the temperature to 70°C After 6 hours, TLC detection showed that the reaction was complete, adding saturated brine, using a rotary evaporator to remove the solvent, extracting the aqueous phase with ethyl acetate (3 × 10mL), combining the organic phases, and extracting with saturated sodium bicarbonate solution (3 × 10mL ), combined the aqueous phases, added 6M HCl solution to adjust the pH to 1, extracted with ethyl acetate (3×20mL), added anhydrous sodium sulfate to dry, filtered after drying, and used a rotary evaporator to remove the solvent to obtain 0.94g of the product. Yield 76%. ¹ H NMR(CDCl ₃ ,400MHz)δ7.23(m,10H),4.65(d,1H,J=11.7Hz),4.42(d,1H,J=11.7Hz),4.20(m,1H),3.12 (m,2H).MS(ESI)m/z255.1[MH] ^- .

Preparation of ((R)-1-((S)-2-(benzyloxy)-3-phenylpropionamido)-3-methylbutyl)boronic acid (compound I-7)

Under nitrogen protection, (S)-2-(benzyloxy)-3-phenylpropionic acid (28 mg, 0.11 mmol) was added to 5 mL of anhydrous dichloromethane, stirred at room temperature, HOBt (18 mg, 0.13 mmol) was added ), continue stirring for 10 minutes and add EDC HCl (25mg, 0.13mmol), then add Example 6 compound III-8-a (50mg, 0.17mmol) and DIPEA (0.04mL, 0.22mmol), after 14 hours, TLC detects It shows that the reaction is complete, and the reaction solution is washed with 5% _NaHCO3 solution, 10% citric acid solution, 5% _NaHCO3 solution and saturated brine, and dried by adding anhydrous sodium sulfate to the organic phase, filtered after drying, and evaporated by rotary evaporation. The solvent was removed by instrumentation, and the resulting product was directly added to 5 mL of methanol, stirred at room temperature, 2-methylpropylboronic acid (22 mg, 0.22 mmol) and 5 mL of n-hexane were added, and then 1M HCl solution (0.33 mL, 0.33 mmol) was added, 5 Hours later, TLC detection showed that the reaction was complete, and the reaction solution was allowed to stand until the layers were separated. The lower layer was washed with n-hexane, dried by adding anhydrous sodium sulfate, filtered after drying, and desolvated by a rotary evaporator, and separated by column chromatography (dichloromethane: Methanol=80:1) 18 mg of white solid was obtained, and the yield was 44%. ¹ H NMR (CD ₃ OD, 400MHz) δ7.24(m, 10H), 4.57(m, 1H), 4.45(m, 1H), 4.32(d, 1H, J=1.3Hz), 3.06(m, 2H ),2.65(t,1H,J=7.6Hz),1.52(m,1H),1.39(t,2H,J=7.7Hz),0.86(d,6H,J=5.5Hz).MS(ESI)m /z368.2[MH] ^- ,382.2[M+CH ₂ -H] ^- ,396.2[M+2CH ₂ -H] ^- .

Embodiment 18: Preparation of 2,2,2-trichloroacetamide (1-naphthylmethyl) ester

Add 1-naphthalenemethanol (1.6g, 10mmol) to 50mL ether, stir at room temperature, add 60% sodium hydride (0.8g, 20mmol), continue stirring for 10 minutes, add trichloroacetonitrile (2.2g, 15mmol), 2 Hours later, TLC detection showed that the reaction was complete. The solvent was removed by a rotary evaporator, and the product was separated by column chromatography (petroleum ether: ethyl acetate = 100:1) to obtain 2 g of the product, with a yield of 66%. ¹ H NMR (CDCl ₃ , 400MHz) δ8.50(s, 1H), 8.07(d, 1H, J=8.0Hz), 7.90(t, 2H, J=7.4Hz), 7.65(d, 1H, J= 6.8Hz), 7.53(m,3H), 5.79(s,2H).

Preparation of (S)-2-(1-naphthylmethoxy)-3-phenylpropanoic acid methyl ester

Add (S)-2-hydroxy-3-phenylpropionic acid methyl ester (1g, 5.5mmol) to 50mL dichloromethane and 50mL n-hexane mixed solvent, stir at room temperature, dropwise add 2,2,2-tris Chloroacetamide (1-naphthylmethyl) ester (2g, 6.6mmol), after the dropwise addition, a catalytic amount of trifluoromethanesulfonic acid was added, and TLC detection after 12 hours showed that the reaction was complete, and the product was obtained by removing the solvent using a rotary evaporator 0.7 g, 40% yield. The reaction product was directly used in the next reaction without purification.

Preparation of (S)-2-(1-naphthylmethoxy)-3-phenylpropionic acid

Add (S)-2-(1-naphthylmethoxy)-3-phenylpropanoic acid methyl ester (0.7g, 2.2mmol) to 20mL dioxane, stir at room temperature, add 26mL of 2M HCl solution, the temperature rises to 70°C, after 4 hours, TLC detection showed that the reaction was complete, adding saturated brine, using a rotary evaporator to remove the solvent, extracting the aqueous phase with ethyl acetate (3 × 10mL), combining the organic phases, and extracting with a saturated sodium bicarbonate solution ( 3×10mL), combined the aqueous phase, added 6M HCl solution to adjust the pH to 1, extracted with ethyl acetate (3×20mL), added anhydrous sodium sulfate to dry, filtered after drying, and used a rotary evaporator to remove the solvent to obtain the product 0.12 g, 18% yield. ¹ H NMR(CDCl ₃ ,400MHz)δ7.45(m,12H),5.12(d,1H,J=11.5Hz),4.84(d,1H,J=11.5Hz),4.31(m,1H),3.12 (m,2H).MS(ESI)m/z305.1[MH] ^- .

Preparation of ((R)-3-methyl-1-((S)-2-(1-naphthylmethoxy)-3-phenylpropionamido)butyl)boronic acid (compound I-8)

Under nitrogen protection, (S)-2-(1-naphthylmethoxy)-3-phenylpropanoic acid (34 mg, 0.11 mmol) was added to 5 mL of anhydrous dichloromethane, stirred at room temperature, and HOBt (18 mg , 0.13mmol), continued to stir for 10 minutes and added EDC·HCl (25mg, 0.13mmol), then added Example 6 compound III-8-a (50mg, 0.17mmol) and DIPEA (0.04mL, 0.22mmol), 14 hours Afterwards, TLC detection showed that the reaction was complete, and the reaction solution was washed with ₅ % NaHCO solution, 10% citric acid solution, ₅ % NaHCO solution and saturated brine, and dried by adding anhydrous sodium sulfate to the organic phase, and filtered after drying. Utilize the rotary evaporator to remove solvent, the obtained product is directly added in 5mL methanol, stir at room temperature, add 2-methylpropylboronic acid (22mg, 0.22mmol) and 5mL n-hexane, then add 1M HCl solution (0.33mL, 0.33mmol ), after 5 hours, TLC detection showed that the reaction was complete, the reaction solution was allowed to stand until the layers were separated, and the lower layer was washed with normal hexane, dried by adding anhydrous sodium sulfate, filtered after drying, and utilized a rotary evaporator to remove the solvent, separated by column chromatography (two Chloromethane:methanol=100:1) to obtain 13 mg of white solid with a yield of 28%. ¹ H NMR (CD ₃ OD, 400MHz) δ7.99(m, 1H), 7.85(t, 2H, J=8.8Hz), 7.35(m, 9H), 4.94(d, 1H, J=9.1Hz), 4.62(s,1H),4.42(m,1H),3.05(m,2H),2.59(t,1H,J=5.6Hz),1.60(m,1H),1.42(t,2H,J=7.2Hz ),0.84(d,6H,J＝6.5Hz).MS(ESI)m/z418.2[MH] ^- ,432.2[M+CH ₂ -H] ^- ,446.2[M+2CH ₂ -H] ^- . .

Embodiment 19: Preparation of 2,2,2-trichloroacetamide (2-pyrazinemethyl) ester

Add 2-hydroxymethylpyrazine (100mg, 0.9mmol) to 5mL ether, stir at room temperature, add 60% sodium hydride (80mg, 2mmol), continue stirring for 10 minutes, then add trichloroacetonitrile (202mg, 1.4mmol) After 2 hours, TLC detection showed that the reaction was complete. The solvent was removed by a rotary evaporator, and the product was separated by column chromatography (petroleum ether: ethyl acetate = 30:1) to obtain 160 mg of the product with a yield of 70%. ¹ H NMR(CDCl ₃ ,400MHz)δ8.79(s,1H),8.56(m,2H),5.51(s,2H).MS(ESI)m/z255.0[M+H] ⁺ .

Preparation of (S)-3-phenyl-2-(2-pyrazinemethoxy)propionic acid methyl ester

Add (S)-2-hydroxy-3-phenylpropionic acid methyl ester (180mg, 1mmol) to 5mL dichloromethane and 5mL n-hexane mixed solvent, stir at room temperature, dropwise add 2,2,2-trichloro Acetamide (2-pyrazinemethyl) ester (316mg, 1.2mmol), after the dropwise addition, a catalytic amount of trifluoromethanesulfonic acid was added, and TLC detection after 12 hours showed that the reaction was complete, and the product was obtained by removing the solvent with a rotary evaporator 156 mg, 57% yield. The reaction product was directly used in the next reaction without purification.

Preparation of (S)-3-phenyl-2-(2-pyrazinemethoxy)propionic acid

Add (S)-3-phenyl-2-(2-pyrazinemethoxy)methyl propionate (156 mg, 0.57 mmol) to 5 mL of dioxane, stir at room temperature, add 6 mL of 2M HCl solution, and the temperature rises to 70°C, TLC detection after 3 hours showed that the reaction was complete, adding saturated saline, using a rotary evaporator to remove the solvent, extracting the aqueous phase with ethyl acetate (3×5mL), combining the organic phases, and extracting with a saturated sodium bicarbonate solution ( 3×5mL), combined the aqueous phase, added 6M HCl solution to adjust the pH to 1, extracted with ethyl acetate (3×5mL), added anhydrous sodium sulfate to dry, filtered after drying, and used a rotary evaporator to remove the solvent to obtain the product 36 mg, 24% yield. ¹ H NMR (CDCl ₃ , 400MHz) δ8.56(m, 5H), 7.85(s, 1H), 7.31(d, 2H, J=6.0Hz), 4.85(s, 1H), 4.25(d, 1H, J＝14.1Hz), 4.07(d, 1H, J＝14.1Hz), 3.24(m, 2H).

Preparation of ((R)-3-methyl-1-((S)-3-phenyl-2-(2-pyrazinemethoxy)propionamido)butyl)boronic acid (compound I-9)

Under nitrogen protection, (S)-3-phenyl-2-(2-pyrazinemethoxy)propionic acid (28 mg, 0.11 mmol) was added to 5 mL of anhydrous dichloromethane, stirred at room temperature, and HOBt ( 18mg, 0.13mmol), continued to stir for 10 minutes and added EDC·HCl (25mg, 0.13mmol), then added Example 6 compound III-8-a (50mg, 0.17mmol) and DIPEA (0.04mL, 0.22mmol), 20 Hours later, TLC detection showed that the reaction was complete, and the reaction solution was washed with 5% _NaHCO3 solution, 10% citric acid solution, 5% _NaHCO3 solution and saturated saline, and dried by adding anhydrous sodium sulfate to the organic phase, and filtered after drying , using a rotary evaporator to remove the solvent, the resulting product was directly added to 5mL of methanol, stirred at room temperature, added 2-methylpropylboronic acid (22mg, 0.22mmol) and 5mL of n-hexane, and then added 1M HCl solution (0.33mL, 0.33 mmol), after 7 hours, TLC detection showed that the reaction was complete, the reaction solution was allowed to stand until the layers were separated, the lower layer was washed with normal hexane, dried by adding anhydrous sodium sulfate, filtered after drying, and utilized a rotary evaporator to remove the solvent, separated by column chromatography ( Dichloromethane:methanol=30:1) to obtain 12 mg of yellow solid with a yield of 29%. ¹ H NMR (CD ₃ OD, 400MHz) δ8.60 (t, 2H, J = 34.3Hz), 8.02 (d, 1H, J = 7.9Hz), 7.61 (m, 6H), 4.39 (d, 1H, J =13.7Hz), 4.22(d,1H,J=14.1Hz),3.72(m,1H),3.20(m,2H),2.91(t,1H,J=12.9Hz),2.02(m,1H), 1.56 (t, 2H, J = 7.2Hz), 0.94 (d, 6H, J = 24.5Hz). MS (ESI) m/z 370.1 [MH] ^- , 398.2 [M+2CH ₂ -H] ^- .

Example 20: Preparation of (S)-2-(benzylamino)-3-phenylpropionic acid

Add (S)-2-amino-3-phenylpropionic acid (165mg, 1mmol) to 10mL water, stir at room temperature, add benzyl chloride (158mg, 1.3mmol) and potassium carbonate (138mg, 1mmol) successively, and then Add benzyl chloride (158mg, 1.3mmol) and potassium carbonate (138mg, 1mmol). After 24 hours, TLC showed that the reaction was complete. Filter and wash with ether. The filter cake was acidified with 6M HCl solution, and a large amount of white solid appeared. Filter to obtain 201mg of the product , yield 79%. ¹ H NMR (D ₂ O-NaOH, 400MHz) δ7.05(m, 10H), 3.48(d, 1H, J=12.7Hz), 3.30(d, 1H, J=12.8Hz), 3.07(t, 1H ,J＝6.8Hz),2.63(m,2H).MS(ESI)m/z254.1[MH] ^- .

Preparation of ((R)-((S)-2-(benzylamino)-3-phenylpropanylamino)-3-methylbutyl)boronic acid (compound I-10)

Under nitrogen protection, (S)-2-(benzylamino)-3-phenylpropionic acid (28 mg, 0.11 mmol) was added to 5 mL of anhydrous dichloromethane, stirred at room temperature, and HOBt (18 mg, 0.13 mmol) was added , add EDC·HCl (25mg, 0.13mmol) after continuing stirring for 10 minutes, then add Example 6 compound III-8-a (50mg, 0.17mmol) and DIPEA (0.04mL, 0.22mmol), after 10 hours TLC detection shows After the reaction is complete, the reaction solution is washed with 5% _NaHCO3 solution, 10% citric acid solution, 5% _NaHCO3 solution and saturated brine, and dried by adding anhydrous sodium sulfate to the organic phase. In addition to solvent, the resulting product was directly added to 5 mL of methanol, stirred at room temperature, 2-methylpropylboronic acid (22 mg, 0.22 mmol) and 5 mL of n-hexane were added, and then 1M HCl solution (0.33 mL, 0.33 mmol) was added for 3 hours After the TLC detection shows that the reaction is complete, the reaction solution is allowed to stand until the layers are separated, the lower layer is washed with n-hexane, dried by adding anhydrous sodium sulfate, filtered after drying, the solvent is removed by a rotary evaporator, and column chromatography is separated (dichloromethane: methanol =50:1) 11 mg of white solid was obtained with a yield of 27%. ¹ H NMR (CD ₃ OD, 400MHz) δ7.23(m, 10H), 3.72(d, 1H, J=13.1Hz), 3.62(d, 1H, J=6.8Hz), 3.37(t, 1H, J =6.9Hz), 3.11(m, 2H), 2.90(t, 1H, J=9.9Hz), 2.03(m, 1H), 1.59(t, 2H, J=8.2Hz), 0.85(d, 6H, J =13.5Hz). MS (ESI) m/z 367.1[MH] ^- , 395.3[M+2CH ₂ -H] ^- .

Example 21: Preparation of (S)-2-((1-naphthylmethyl)amino)-3-phenylpropionic acid

Add (S)-2-amino-3-phenylpropionic acid (1g, 6mmol) to 50mL water, stir at room temperature, add 1-chloromethylnaphthalene (1.3g, 7.5mmol) and potassium carbonate (0.9g, 6mmol) ), add 1-chloromethylnaphthalene (1.3g, 7.5mmol) and salt of wormwood (0.9g, 6mmol) after 1 hour, after 30 hours, TLC detects and shows that the reaction is complete, filters and washes with ether, and the HCl of filter cake utilizes 6M The solution was acidified, a large amount of white solid appeared, and 0.69 g of the product was obtained by filtration, with a yield of 38%. ¹ H NMR (CD ₃ OD, 400MHz) δ7.60(m, 12H), 4.75(d, 1H, J=13.2Hz), 4.59(d, 1H, J=13.7Hz), 4.09(m, 1H), 3.24(m,2H).MS(ESI)m/z306.2[M+H] ⁺ .

Preparation of ((R)-3-methyl-1-((S)-2-((1-naphthylmethyl)amino)-3-phenylpropionamido)butyl)boronic acid (compound I-11)

Under nitrogen protection, (S)-2-((1-naphthylmethyl)amino)-3-phenylpropionic acid (34 mg, 0.11 mmol) was added to 5 mL of anhydrous dichloromethane, stirred at room temperature, and HOBt was added (18mg, 0.13mmol), continue stirring for 10 minutes and add EDC·HCl (25mg, 0.13mmol), then add Example 6 compound III-8-a (50mg, 0.17mmol) and DIPEA (0.04mL, 0.22mmol), After 15 hours, TLC detection showed that the reaction was complete, and the reaction solution was washed with ₅ % NaHCO solution, 10% citric acid solution, ₅ % NaHCO solution and saturated brine, and dried by adding anhydrous sodium sulfate to the organic phase. Filtrate, use a rotary evaporator to remove the solvent, add the resulting product directly to 5 mL of methanol, stir at room temperature, add 2-methylpropylboronic acid (22 mg, 0.22 mmol) and 5 mL of n-hexane, and then add 1M HCl solution (0.33 mL, 0.33mmol), after 5 hours, TLC detection showed that the reaction was complete, the reaction solution was allowed to stand until the layers were separated, the lower layer was washed with n-hexane, dried by adding anhydrous sodium sulfate, filtered after drying, and the solvent was removed by a rotary evaporator, and column chromatography separated (dichloromethane:methanol=50:1) to obtain 13 mg of white solid with a yield of 28%. ¹ H NMR (CD ₃ OD, 400MHz) δ7.53(m, 12H), 4.15(m, 2H), 3.72(m, 1H), 3.02(d, 2H, J=7.2Hz), 2.61(t, 1H ,J=7.4Hz),1.48(m,1H),1.08(t,2H,J=6.9Hz),0.83(d,6H,J=7.5Hz).MS(ESI)m/z417.2[MH] ^- ,431.2[M+CH ₂ -H] ^- ,445.3[M+2CH ₂ -H] ^- .

Example 22: Preparation of (S)-3-phenyl-2-((2-pyrazinemethyl)amino)propionic acid

Add (S)-2-amino-3-phenylpropionic acid (83mg, 0.5mmol) to 5mL water, stir at room temperature, add 2-chloromethylpyrazine (75mg, 0.6mmol) and potassium carbonate (69mg, 0.5 mmol), add 2-chloromethylpyrazine (75mg, 0.6mmol) and potassium carbonate (69mg, 0.5mmol) after 1 hour, after 26 hours, TLC detection shows that the reaction is complete, filters and washes with ether, and the filter cake utilizes 6M The HCl solution was acidified, a large amount of light yellow solid appeared, and 103 mg of the product was obtained by filtration, with a yield of 80%. ¹ HNMR (D ₂ O–NaOH, 400MHz)δ8.08(m,3H),6.86(m,5H),3.49(d,1H,J=14.4Hz),3.33(d,1H,J=14.6Hz) ,2.90(m,1H),2.48(m,2H).MS(ESI)m/z256.0[MH] ^- .

((R)-3-methyl-1-((S)-3-phenyl-2-((2-pyrazinemethyl)amino)propionamido)butyl)boronic acid (Compound I-12) preparation

Under nitrogen protection, (S)-3-phenyl-2-((2-pyrazinemethyl)amino)propionic acid (28 mg, 0.11 mmol) was added to 5 mL of anhydrous dichloromethane, stirred at room temperature, added HOBt (18mg, 0.13mmol), continue to stir for 10 minutes and then add EDC·HCl (25mg, 0.13mmol), then add Example 6 compound III-8-a (50mg, 0.17mmol) and DIPEA (0.04mL, 0.22mmol) After 17 hours, TLC detection showed that the reaction was complete, and the reaction solution was washed with ₅ % NaHCO solution, 10% citric acid solution, ₅ % NaHCO solution and saturated brine, and dried by adding anhydrous sodium sulfate to the organic phase. After filtering, use a rotary evaporator to remove the solvent, and the resulting product is directly added to 5 mL of methanol, stirred at room temperature, added 2-methylpropylboronic acid (22 mg, 0.22 mmol) and 5 mL of n-hexane, and then added 1M HCl solution (0.33 mL , 0.33mmol), after 6 hours, TLC detection showed that the reaction was complete, the reaction solution was allowed to stand until the layers were separated, the lower layer was washed with n-hexane, dried by adding anhydrous sodium sulfate, filtered after drying, and the solvent was removed by rotary evaporator, column chromatography Separation (dichloromethane:methanol=50:1) yielded 11 mg of a yellow solid with a yield of 27%. ¹ H NMR(CD ₃ OD,400MHz)δ8.09(s,1H),7.21(s,1H),6.95(s,1H),6.13(m,5H),4.04(d,1H,J＝13.4Hz ),3.81(d,1H,J=3.2Hz),3.54(m,1H),3.15(m,2H),2.80(t,1H,J=8.9Hz),2.02(m,1H),1.58(t ,2H, J=10.2Hz),0.89(d,6H,J=7.3Hz).MS (ESI) m/z369.0[MH] ^- ,397.0[M+2CH ₂ -H] ^- .

Embodiment 23: Determination experiment of proteasome inhibitory activity of the compound of the present invention

In this example, the 20S proteasome kit purchased from Chemicon (Chemicon, USA) was used for testing. The basic principle of this test method is that 20S proteasome hydrolyzes the fluorescent substrate Suc-LLVY-AMC (Suc-Leu-Leu-Val-Tyr-AMC, Suc is succinyl, AMC is 7-amide-4-methylcoumarin ), release AMC with fluorescence, by changing the concentration of the compound to be tested, use SpectraMax M5 microplate reader (380nm/460nm) to measure the fluorescence value of releasing AMC under different concentrations, thereby judging the compound’s effect on 20S proteasome chymotrypsin-like The degree of inhibition of the active site (CT-L), using the GraphPad Prism software to calculate the _IC50 value of the inhibition of the compound on the 20S proteasome chymotrypsin-like active site; Table 1 shows the effect of some compounds on the 20S proteasome chymotrypsin-like active site _IC50 values for spot inhibition eg.

Table 1. Inhibition of the tested compounds on the chymotrypsin-like active site of the human 20S proteasome

• _IC50 values are the mean of at least two independent experiments.

Claims (9)

1. the boronic acid compound of formula I structure, in: X is O or NH; R is aryl or heteroaryl; n is 0 or 1. 2. boric acid compound according to claim 1, is characterized in that, described compound is the compound with following structure, 3. boric acid compound according to claim 1, is characterized in that, described compound is the compound with following structure, 4. boric acid compound according to claim 2, is characterized in that, described compound is the compound with following structure, 5. boric acid compound according to claim 2, is characterized in that, described compound is the compound with following structure, 6. boric acid compound according to claim 3, is characterized in that, described compound is the compound with following structure, 7. boric acid compound according to claim 3, is characterized in that, described compound is the compound with following structure, 8. Use of the compound according to any one of claims 1-7 in the preparation of proteasome inhibitors. 9. The use according to claim 8, wherein said proteasome inhibitor is a drug for treating diseases related to proteasome.