CN113121600A

CN113121600A - Remdesivir dimer, preparation method and application thereof

Info

Publication number: CN113121600A
Application number: CN202110351812.0A
Authority: CN
Inventors: 姜志宏; 周小波; 李润峰
Original assignee: Macau University of Science and Technology
Current assignee: Macau University of Science and Technology
Priority date: 2021-03-31
Filing date: 2021-03-31
Publication date: 2021-07-16

Abstract

The invention belongs to the technical field of drug synthesis, and discloses a Remdesivir dimer, and a preparation method and application thereof. The structural formula of the Remdesivir dimer is shown as a formula (I), the Remdesivir dimer provided by the invention is short in preparation route, mild in reaction condition, simple in post-treatment and high in yield up to 75%, can contribute to development of virus medicines and research on clinical application of Remdesivir, and is a potential medicine.

Description

Remdesivir dimer, preparation method and application thereof

Technical Field

The invention belongs to the technical field of drug synthesis, and particularly relates to a Remdesivir dimer and a preparation method and application thereof.

Background

Remdesivir is a nucleoside antiviral drug, is a nucleoside RNA-dependent RNA polymerase (RdRp) competitive inhibitor, and a nucleotide triphosphate product Remdesivir-TP thereof can compete with RdRp for a substrate ATP, so that virus vRNA synthesis can be interfered. Remdesivir is effective against a variety of coronavirus infections, including the atypical pneumonia coronavirus (SARS-CoV) and the middle east respiratory syndrome associated coronavirus (MERS-CoV), and has been used in clinical trials for the treatment of novel coronavirus pneumonia (covid-19) in several countries of the world. However, the curative effect and safety of the Remdesivir are not completely proved, so that the synthesis of the Remdesivir analogue is an important matter for researching the toxicity and the side effects of the Remdesivir analogue, and has important reference significance for the development of virus medicines and the clinical application of the Remdesivir. However, at present, the synthesis of Remdesivir analogues is less, and the problems of complex synthesis method, complicated post-treatment process, low yield and the like exist.

Therefore, there is a need to provide Remdesivir analogs, and methods of preparation that are simple and easy to perform.

Disclosure of Invention

The present invention is directed to solving at least one of the problems of the prior art described above. Therefore, the Remdesivir dimer provided by the invention can contribute to development of virus drugs, and the preparation method is simple and does not need a complex post-treatment process.

The structural formula of the Remdesivir dimer is shown as a formula (I),

the invention also provides a preparation method of the Remdesivir dimer,

specifically, the preparation method of the Remdesivir dimer comprises the following steps:

(1) dissolving the compound 1, diisocyanate and an adsorbing material in a first reaction solvent, and stirring for reaction to generate an intermediate 2; the structure of the compound 1 is shown as a formula (II), and the structure of the intermediate 2 is shown as a formula (III);

(2) and dissolving the intermediate 2 in a second reaction solvent, adding acid, and reacting to obtain the Remdesivir dimer.

Preferably, in step (1), the adsorption material is at least one of molecular sieve, silica gel or activated carbon.

Further preferably, the adsorption material is a 4A molecular sieve. The 4A molecular sieve can dry trace moisture in the solvent, provide proper pores for reaction and promote the reaction.

Preferably, in the step (1), the diisocyanate is a diisocyanate containing a methylene structure; further preferably, in the step (1), the diisocyanate is one selected from the group consisting of hexamethylene diisocyanate, pentamethylene diisocyanate, tetramethylene diisocyanate, trimethylene diisocyanate, octamethylene diisocyanate and decamethylene diisocyanate.

Preferably, in step (1), the first reaction solvent is an organic solvent; further preferably, the organic solvent is at least one selected from dichloromethane, chloroform, tetrahydrofuran, dimethylsulfoxide, or acetonitrile.

Preferably, in the step (1), the temperature of the stirring reaction is 20-50 ℃, and the time of the stirring reaction is 5-20 h; further preferably, in the step (1), the temperature of the stirring reaction is 20-40 ℃, and the time of the stirring reaction is 8-15 h.

Preferably, in the step (2), the second reaction solvent is selected from at least one of methanol, ethanol, N-dimethylformamide or N, N-dimethylacetamide.

Preferably, in the step (2), the acid is selected from at least one of hydrochloric acid, formic acid, acetic acid or trifluoroacetic acid.

Preferably, in the step (2), the reaction temperature is 20-70 ℃, and the reaction time is 5-20 h; further preferably, in the step (2), the reaction temperature is 20-60 ℃, and the reaction time is 8-15 h.

A medicament comprising said Remdesivir dimer.

Preferably, the medicament also comprises a pharmaceutically acceptable carrier or auxiliary material; preferably includes at least one of a diluent, an absorbent, a humectant, a disintegrant, a lubricant, a disintegration inhibitor, and a binder; further preferably, the binder includes at least one of starch, dextrin, calcium sulfate, lactose, mannitol, sucrose, sodium chloride, glucose, urea, calcium carbonate, kaolin, microcrystalline cellulose, aluminum silicate, water, glycerin, polyethylene glycol, ethanol, propanol, starch slurry, dextrin, syrup, honey, glucose solution, gum arabic slurry, gelatin slurry, sodium carboxymethylcellulose, shellac, methyl cellulose, potassium phosphate, polyvinylpyrrolidone, dried starch, alginate, agar powder, brown algae starch, sodium bicarbonate and citric acid, calcium carbonate, polyoxyethylene, sorbitol fatty acid ester, sodium dodecyl sulfonate, methyl cellulose, ethyl cellulose, sucrose, tristearin, cacao butter, hydrogenated oil, talc, silicon dioxide, corn starch, stearate, boric acid, liquid paraffin, and polyethylene glycol.

Preferably, the dosage form of the medicament is selected from oral dosage forms; further preferably, the oral dosage form is at least one of tablets, capsules, granules and dropping pills.

Preferably, the medicament further comprises other therapeutic ingredients.

The Remdesivir structure has a plurality of active reaction sites, such as amino groups, hydroxyl groups, phosphate ester bonds and the like, so to obtain the dimer provided by the invention, the activity of the plurality of reaction sites in the Remdesivir structure needs to be controlled, the reaction steps are increased invisibly, and the post-treatment purification process is also increased. It was also found during the course of the conditional screening that if highly reactive reagents are used, such as the diacylchloride reagent, dimers cannot be obtained but the diacylchloride reacts with the nitrogen on the base exocyclic amine to form the product diimide. The Remdesivir dimer can be prepared in an extremely short reaction route by selecting reactants and controlling reaction conditions. And the reaction condition is mild, complex post-treatment is not needed, and the yield is high.

Compared with the prior art, the invention has the following beneficial effects:

the Remdesivir dimer provided by the invention has the advantages of short preparation route, mild reaction conditions, simple post-treatment and yield up to 75%, can be beneficial to development of virus medicines and research on clinical application of Remdesivir, and is also a potential medicine.

Drawings

FIG. 1 is a process flow diagram for the preparation of Remdesivir dimer in example 1.

Detailed Description

In order to make the technical solutions of the present invention more apparent to those skilled in the art, the following examples are given for illustration. It should be noted that the following examples are not intended to limit the scope of the claimed invention.

All organic reagents in the following examples were purchased from shanghai jiding chemistry. The conditions for the mass spectrometry test were: bruker Ascend 600 NMR spectrometer running XWINNMR software package (¹The H NMR was 600MHz and the resulting mixture was,¹³c NMR 150MHz) was used for NMR experiments. High resolution MS mass spectra were acquired on a mass spectrometer in positive or negative ion mode Agilent 6230 precision mass time of flight mass spectrometer (usa) equipped with an electrospray ion source (ESI). Column Chromatography (CC) separation was performed using silica gel 60(200-300 mesh, Bio-Gene Technology Ltd. hong Kong).

Other starting materials, reagents or equipment may be obtained from conventional commercial sources or may be obtained by known methods in the art, unless otherwise specified.

Example 1

The structural formula of the Remdesivir dimer is shown as a formula (I),

the preparation method of the Remdesivir dimer comprises the following steps (the process flow diagram is shown in figure 1):

(1) dissolving the compound 1(15.5mg, 0.024mmol, the structural formula is shown as (II)) in anhydrous DCM (1mL), adding hexamethylene dinitrate (1.0uL, 0.006mmol) and 4A molecular sieve (4A MS), stirring the mixture at 25 ℃ overnight (12h), filtering, spin-drying, and separating the mixture by column chromatography to obtain an intermediate 2(6.3mg, the yield is 72%, the structural formula is shown as (III)), and the intermediate 2 is white powder.

The nuclear magnetic characterization results and mass spectra of intermediate 2 were as follows:¹H NMR(600MHz,CDCl₃)δ10.06(s,2H),9.48(s,2H),7.33(s,2H),7.28-7.25(m,2H),7.20-7.18(m,4H),7.12(t,J＝7.4Hz,2H),6.97(d,J＝4.2Hz,2H),5.37(d,J＝4.2Hz,2H),4.96-4.95(m,2H),4.60-4.58(m,2H),4.40-4.36(m,2H),4.34-4.31(m,2H),4.14-4.11(m,2H),4.03-4.00(m,2H),3.98-3.94(m,2H),3.93-3.90(m,2H),3.65-3.61(m,2H),3.35-3.33(m,2H),1.80-1.73(m,4H),1.76(s,6H),1.66-1.59(m,4H),1.62(s,6H),1.56-1.51(m,2H),1.48-1.44(m,2H),1.34(d,J＝7.0Hz,6H),1.31-1.26(m,12H),0.84(t,J＝7.5Hz,12H)；¹³C NMR(150MHz,CDCl₃)δ173.6,173.5,155.0,151.6,150.6,150.6,144.8,129.6,125.0,123.9,120.2,120.2,117.2,116.1,115.5,113.2,102.8,83.9,83.4,83.4,81.9,81.4,67.5,65.4,65.4,50.2,40.2,29.7,26.2,25.8,25.4,23.1,23.1,21.0,21.0,10.9,10.9；HRMS(ESI):m/z for C₆₈H₉₀N₁₄O₁₈P₂calcd 1452.6033,found 1453.7374 for[M+H]⁺,1475.7235 for[M+Na]⁺,727.4002 for[M+2H]²⁺。

(2) intermediate 2(12.9mg, 0.009mmol) was dissolved in MeOH (1mL), 100. mu.L of concentrated HCl was added, the mixture was stirred overnight (12h) at room temperature (25 deg.C), filtered, spun dry, and the mixture was isolated by column chromatography to give Remdesivir dimer (9.1mg, 75%) as a white powder.

The nuclear magnetic characterization results and mass spectra of Remdesivir dimer are as follows:¹H NMR(600MHz,CD₃OD)δ8.2(s,2H),7.39-7.26(m,6H),7.20-7.14(m,8H),4.76-4.74(m,2H),4.42-4.38(m,4H),4.33-4.37(m,2H),4.30-4.27(m,2H),4.18-4.15(m,2H),4.03-4.00(m,2H),3.95-3.92(m,2H),3.88-3.86(m,2H),3.40-3.38(m,4H),1.70-1.65(m,2H),1.55-1.48(m,2H),1.47-1.39(m,2H),1.33-1.28(m,10H),1.15(d,J＝6.2Hz,6H),0.85(t,J＝7.5Hz,12H)；¹³C NMR(150MHz,CD₃OD)δ173.6,169.8,153.9,150.7,150.7,149.5,140.4,129.5,129.4,124.7,120.0,119.9,115.5,114.7,113.6,106.1,83.0,82.9,79.4,74.7,70.1,67.8,66.7,65.5,63.3,51.4,50.1,50.0,40.3,40.3,39.6,29.0,26.1,23.8,22.8,22.8,19.2,19.2,10.0,9.9；HRMS(ESI):m/z for C₆₂H₈₂N₁₄O₁₈P₂ calcd 1372.5407,found 1373.5466 for[M+H]⁺,1395.5278 for[M+Na]⁺,687.2726 for[M+2H]²⁺。

product effectiveness testing

The Remdesivir dimer prepared in example 1 was evaluated for drug toxicity.

(1) Experimental Material

Vero E6 (Vero green monkey kidney cells) cells were purchased from ATCC in the USA.

(2) Toxicity test of Remdesivir dimer on cells

Dissolving Remdesivir and Remdesivir dimer prepared in example 1 in DMSO (dimethyl sulfoxide), ultrasonically filtering, adding DMEM culture solution to dilute to 5mg/mL, filtering with 0.22 μm filter membrane, preserving at 4 ℃, discarding culture solution of 96-well plate monolayer Vero E6 cells, adding 100 μ L/well of drug with different dilutions, adding equivalent volume of DMEM culture solution into normal cell control well, and culturing at 37 ℃ and 5% CO₂Under the condition, after culturing for 72 hours, observing the cytopathic Condition (CPE) caused by the medicine, and determining CC₀(maximum non-toxic concentration of drug to cells). The test results are shown in Table 1.

TABLE 1 inhibition of CPE-causing Vero E6 by Remdesivir dimer

Claims

The Remdesivir dimer is characterized in that the structural formula of the Remdesivir dimer is shown as a formula (I),
2. the method of producing a remdevivir dimer as defined in claim 1, comprising the steps of:

(1) dissolving the compound 1, diisocyanate and an adsorbing material in a first reaction solvent, and stirring for reaction to generate an intermediate 2; the structure of the compound 1 is shown as a formula (II), and the structure of the intermediate 2 is shown as a formula (III);

(2) and (2) dissolving the intermediate 2 prepared in the step (1) in a second reaction solvent, adding acid, and reacting to prepare the Remdesivir dimer.
3. The preparation method according to claim 2, wherein in the step (1), the adsorbing material is at least one of molecular sieve, silica gel or activated carbon.
4. The method of claim 3, wherein the adsorbent material is a 4A molecular sieve.
5. The process according to claim 2, wherein in the step (1), the diisocyanate is a methylene group-containing diisonitrile ester.
6. The method according to claim 2, wherein in the step (1), the first reaction solvent is at least one selected from dichloromethane, chloroform, tetrahydrofuran, dimethylsulfoxide and acetonitrile.
7. The preparation method according to claim 2, wherein in the step (1), the temperature of the reaction is 20 to 50 ℃, and the time of the reaction is 5 to 20 hours; in the step (2), the reaction temperature is 20-70 ℃, and the reaction time is 5-20 h.
8. The method according to claim 2, wherein in the step (2), the second reaction solvent is at least one selected from methanol, ethanol, N-dimethylformamide and N, N-dimethylacetamide.
9. The production method according to claim 2, wherein in the step (2), the acid is at least one selected from the group consisting of hydrochloric acid, formic acid, acetic acid and trifluoroacetic acid.
10. A medicament comprising the Remdesivir dimer of claim 1.