CN115819423A

CN115819423A - ProTAC compound of Reidesciclovir or intermediate thereof, preparation method thereof and application of anti-EV 71

Info

Publication number: CN115819423A
Application number: CN202211520619.6A
Authority: CN
Inventors: 周海兵; 吴叔文; 麻笑雨
Original assignee: Wuhan University WHU
Current assignee: Wuhan University WHU
Priority date: 2022-11-29
Filing date: 2022-11-29
Publication date: 2023-03-21

Abstract

The invention discloses a ProTAC compound of Reidesciclovir or an intermediate thereof, a preparation method thereof and application of anti-EV 71, belonging to the technical field of medicines. The PROTAC compound is obtained by connecting Rudexilvir or an intermediate GS-441524 thereof with an E3ligase ligand through a Linker, has the activity of resisting EV71 viruses, can be developed as a novel medicament for resisting the EV71 viruses, and has wide application prospect.

Description

ProTAC compound of Reidesciclovir or intermediate thereof, preparation method thereof and application of anti-EV 71

Technical Field

The invention belongs to the technical field of medicines, and relates to a ProTAC compound based on Reidesciclovir or an intermediate GS-441524 thereof, a preparation method thereof and application of the compound in resisting EV71 virus.

Background

Enterovirus type 71 (EV 71) belongs to members of the genus Enterovirus in the picornavirus family, and is a positive-sense single-stranded RNA virus comprising an open reading frame encoding four cap membrane proteins (VP 1-4) and seven nonstructural proteins (2A, 2B, 2C, 3A, 3B, 3C, and 3D). The 3D (also known as 3 dppci 1) protein, as the viral RNA-dependent RNA polymerase (RdRp), plays a major role in the negative strand synthesis of the virus and uridylation of some proteins. EV71 caused epidemic and sporadic outbreaks of hand-foot-and-mouth disease (HFMD) over the past 20 years, and may also induce severe neurological/cardiopulmonary complications such as aseptic meningitis, acute flaccid paralysis, encephalitis, myocarditis, pulmonary edema, and hemorrhage, leading to death, making it one of the most pathogenic enteroviruses. Since the large-scale outbreak in 2008, hand-foot-and-mouth disease has spread widely in china, with over 200 million cases per year, and most patients are children.

Enterovirus type 71 (EV 71), one of the viruses that currently still poses a threat to global health, has no FDA approval for specific drugs for clinical use. In response to the threat of EV71 to public health safety, vaccine prophylaxis and symptomatic treatment are the only clinical countermeasures to exist. However, due to genomic mutations, it is useless and ineffective for the infected, and there is an urgent need to develop new antiviral drugs to treat ER-a71 infections.

Disclosure of Invention

The invention aims to overcome the defects of the existing medicines and provide a ProTAC compound of the Reidesciclovir or the intermediate GS-441524 thereof, wherein the ProTAC compound of the Reidesciclovir or the GS-441524 has the activity of resisting EV71 virus, can be developed as a novel medicament for resisting EV71 virus, and has wide application prospect.

Another object of the present invention is to provide a process for preparing ProTAC compounds of Rudexilvir or GS-441524.

In order to achieve the purpose, the technical scheme adopted by the invention is as follows:

in a first aspect, the present invention provides a ProTAC compound of Rudexilvir or GS-441524. Wherein the structure of the ProTAC compound of the Rideciclovir is shown as a general formula (I); the structure of the PROTAC compound of GS-441524 is shown as a general formula (II) or (III).

In the above formula, the E3 ligand is a small molecule protease ligand having ubiquitination function and encoded by VHL (von Hippel-Lindau ), CRBN (Cereblon) or RNF114 gene, and specifically includes

Furthermore, the E3 ligand may be a hydrophobic tag small molecule ligand, including

Linker is a linking group, representing an-alkylene or-alkoxy group, wherein the-alkylene or-alkoxy group is optionally a straight or branched chain alkylene or alkoxy group interrupted one or more times by one or more groups selected from, for example, - (CH) ₂ ) _n -、-(CH ₂ ) _n CO-、-NR ₁ (CH ₂ ) _n CO-、-NR ₂ (CH ₂ ) _n -、-(OCH ₂ CH ₂ O) _n -、-(CH ₂ CH ₂ O) _n -、-(OCH ₂ CH ₂ OCH ₂ ) _n -、-(CH ₂ CH ₂ OCH ₂ ) _n -、-(CH ₂ CH ₂ OCH ₂ CH ₂ ) _n -, heteroarylene group or any combination thereof, wherein n represents a natural number from 1 to 20, R ₁ 、R ₂ Each independently of the other is selected from H or C _1-10 An alkyl group.

Preferably, the ridciclovir or GS-441524 type PROTAC compounds provided by the present invention include, but are not limited to, the compounds shown in table 1 below:

TABLE 1

In a second aspect, the invention provides an application of any one of the procac compounds of reiciclovir or GS-441524 or a pharmacologically or physiologically acceptable salt thereof as a degradation agent of an RNA-dependent RNA polymerase inhibitor in preparing an anti-EV 71 medicament.

In a third aspect, the present invention provides a pharmaceutical composition or an RNA-dependent RNA polymerase inhibitor against EV71, comprising the aforementioned prodec compound of redciclovir or GS-441524 or a pharmacologically or physiologically acceptable salt thereof, and further comprising a pharmaceutically acceptable carrier or excipient.

In a fourth aspect, the invention provides a preparation method of the ProTAC compound of the Reidesciclovir or GS-441524, which is characterized in that the ProTAC compound of the Reidesciclovir or GS-441524 is divided into a ProTAC compound of series I Reidesciclovir, a PROTAC compound of series II GS-441524, a PROTAC compound of series III GS-441524, a PROTAC compound of series IV GS-441524 and a PROTAC compound of series V-441524.

Wherein, the PROTAC compound of the series I Ruidexilvir is a compound shown in a general formula (I), linker is an alkylene chain or an alkoxy chain with n =6-20, and E3ligase is the VHL with methyl.

PROTAC compounds of series II GS-441524 are compounds represented by the general formula (II), linker is an-alkylene chain with n =6-10, and E3 ligand is the above-mentioned methyl-bearing VHL.

The PROTAC compound of the series III GS-441524 is a compound shown as a general formula (III), linker is an alkylene or alkoxy chain with n =6-20, and E3 ligand is the methyl-bearing VHL or trans-methyl-bearing VHL.

The PROTAC compound of the series IV GS-441524 is a compound shown as a general formula (III), a Linker is an alkylene chain with n =6-20, and E3 ligand is the lenalidomide, the methyllenalidomide or the benzodione piperidine.

The PROTAC compound of the series V GS-441524 is a compound shown as a general formula (III), a Linker is an alkylene chain with n =6-11, and E3 ligand is Boc lysine, 9-aminofluorene and RNF-114 ligand.

The preparation method of the PROTAC compound of series I Ruixiwei comprises the following steps:

reacting a VHL derivative with an alkanoic acid at the end and an alkyl chain or an alkoxy chain as a connecting chain with Reidesciclovir in a solvent under the conditions of HATU (O- (7-azabenzotriazole-1-yl) -N, N, N ', N' -tetramethylurea hexafluorophosphate) and DIPEA (N, N-diisopropylethylamine) to obtain a PROTAC compound of the Reidesciclovir series I; the solvent is preferably anhydrous DCM (dichloromethane) or DMF (N, N-dimethylformamide) or a mixed solution of the two, the reaction is preferably carried out at 0-25 ℃, and the reaction time is preferably 1-12 h.

Synthetic route of PROTAC compound of series I Ruixiwei

A preparation method of PROTAC compounds of series II GS-441524 comprises the following steps:

reacting a VHL derivative with an alkanoic acid at the tail end and an alkyl chain as a connecting chain with GS-441524 in a solvent under the conditions of HATU and DIPEA to obtain a PROTAC compound of a series II GS-441524; the solvent is preferably DCM or DMF or a mixed solvent of the DCM and the DMF, the reaction is preferably carried out at 0-25 ℃, and the reaction time is preferably 1-12 h.

Synthetic route of PROTAC compound of series II GS-441524

The preparation method of the PROTAC compound of the series III GS-441524 comprises the following 3 methods:

(1) GS-441524 reacts with 2, 2-dimethoxypropane and concentrated sulfuric acid in acetone to obtain a key intermediate protected by 2'-OH and 3' -OH, wherein the reaction time is preferably 1h-3h. The intermediate and the single alkyl chain VHL derivative without tert-butyl ester are then reacted in a solvent under the conditions of EDCI (1-ethyl-3 (3-dimethylpropylamine) carbodiimide), DMAP (4-dimethylaminopyridine) to obtain the 2'-OH,3' -OH protected PROTAC compound GS-441524, wherein the solvent is preferably DCM or DMF or a mixed solvent of the two, the reaction is preferably carried out at 0-25 ℃, and the reaction time is preferably 1-12 h. Finally, removing the protecting group under the acidic condition of 0-25 ℃ to obtain the PROTAC compound of the series III GS-441524.

Synthesis route of PROTAC compound of series III (1) GS-441524

(2) The 2'-OH,3' -OH protected GS-441524 key intermediate and the single alkyl chain VHL derivative with tert-butyl ester removed are then reacted in a solvent, preferably DCM or DMF or a mixed solvent thereof, preferably at 0-25 ℃ for 1-12 h under EDCI and DMAP conditions to obtain the 2'-OH,3' -OH protected GS-441524 PROTAC compound. Finally, removing the protecting group under the acidic condition of 0-25 ℃ to obtain the PROTAC compound of the series III GS-441524.

Synthesis route of PROTAC compound of series III (2) GS-441524

(3) The method comprises the steps of dissolving a methyl VHL derivative with a bromine atom at the end and a single alkyl chain as a connecting chain, a GS-441524 key intermediate protected by 2'-OH,3' -OH, potassium carbonate and potassium iodide in a solvent, and carrying out reflux reaction at 60-85 ℃ to obtain a PROTAC compound of the GS-441524 protected by 2'-OH,3' -OH, wherein the solvent is preferably acetonitrile or DMF, and the reaction time is preferably 6-12h. Finally, removing the protecting group under the acidic condition of 0-25 ℃ to obtain the PROTAC compound of the series III GS-441524.

Synthesis route of PROTAC compound of series III (3) GS-441524

The preparation of ProTAC compounds of series IV GS-441524 comprises the following 2 methods:

(1) The CRBN derivative with an alkanoic acid at the end and an alkoxy connecting chain reacts with the GS-441524 intermediate protected by 2'-OH,3' -OH in a solvent under the conditions of EDCI and DMAP to obtain the PROTAC compound of the GS-441524 protected by 2'-OH,3' -OH, wherein the solvent is preferably DCM, DMF or a mixed solvent of the DCM and the DMF, the reaction is preferably carried out at 0-25 ℃, and the reaction time is preferably 1-12 h. Finally, removing the protecting group under the acidic condition of 0-25 ℃ to obtain the PROTAC compound of the series IV GS-441524.

Synthetic route of PROTAC compound of series IV GS-441524

(2) The benzodione piperidine of which the tail end is an alkanoic acid and the connecting chain is an alkoxy chain reacts with the GS-441524 intermediate protected by 2'-OH,3' -OH in a solvent under the conditions of EDCI and DMAP to obtain the PROTAC compound of the GS-441524 protected by 2'-OH,3' -OH, wherein the solvent is preferably DCM, DMF or a mixed solvent of DCM and DMF, the reaction is preferably carried out at 0-25 ℃, and the reaction time is preferably 1h-12h. Finally, removing the protecting group under the acidic condition of 0-25 ℃ to obtain the PROTAC compound of the series III GS-441524.

Synthetic route of PROTAC compound of series IV GS-441524

The preparation method of the PROTAC compound of the series V GS-441524 comprises the following 3 methods:

(1) The long-chain alkanoic acid with the terminal Boc (tert-butyloxycarbonyl) protected amino group reacts with the 2'-OH,3' -OH protected GS-441524 intermediate in a solvent under the conditions of EDCI and DMAP to obtain the 2'-OH,3' -OH protected GS-441524 Linker conjugate compound, wherein the solvent is preferably DCM, DMF or a mixed solvent of DCM and DMF, the reaction is preferably carried out at 0-25 ℃, and the reaction time is preferably 1-12 h. Then removing protecting group under the acidic condition of 0-25 ℃ to obtain GS-441524 which is connected with a long alkyl chain and the tail end of which is naked, and finally reacting with lysine protected by Boc to obtain series V GS-441524 PROTAC compounds.

Synthetic route of PROTAC compound of series V GS-441524

(2) The 9-aminofluorene derivative with an alkanoic acid at the end and an alkoxy connecting chain and the GS-441524 intermediate protected by 2'-OH,3' -OH react in a solvent under the conditions of EDCI and DMAP to obtain the PROTAC compound of the GS-441524 protected by 2'-OH,3' -OH, wherein the solvent is preferably DCM, DMF or a mixed solvent of DCM and DMF, the reaction is preferably carried out at 0-25 ℃, and the reaction time is preferably 1-12 h. Finally, removing the protecting group under the acidic condition of 0-25 ℃ to obtain the PROTAC compound of the series V GS-441524.

Synthetic route of PROTAC compound of series V GS-441524

(3) RNF-114 with an alkanoic acid at the end and an alkoxy connecting chain and a GS-441524 intermediate protected by 2'-OH and 3' -OH react in a solvent under the conditions of EDCI and DMAP to obtain the PROTAC compound of the GS-441524 protected by 2'-OH and 3' -OH, wherein the solvent is preferably DCM, DMF or a mixed solvent of DCM and DMF, the reaction is preferably carried out at 0-25 ℃, and the reaction time is preferably 1h-12h. Finally, removing the protecting group under the acidic condition of 0-25 ℃ to obtain the PROTAC compound of the series V GS-441524.

Synthetic route of PROTAC compound of series V GS-441524

The preparation method of the series I PROTAC compound has the following steps that the ratio of the quantities of the Redexilvir, the VHL derivative, the HATU and the DIPEA is 1.1.

The ratio of the amounts of GS-441524, VHL derivative, HATU and DIPEA in the above-described process for the preparation of series II procac compounds is 1.1.

In the preparation methods (1), (2) and (3) of the PROTAC compound of the series III, the mass ratio of GS-441524, 2-dimethoxypropane and concentrated sulfuric acid is 1; the mass ratio of the 2'-OH,3' -OH protected GS-441524 intermediate, VHL derivative, EDCI, DMAP was 1; the volume ratio of HCl to THF was 0.2.

In the preparation method (1) of the series IV of PROTAC compounds, the mass ratio of the 2'-OH,3' -OH protected GS-441524 intermediate, the CRBN derivative, EDCI and DMAP is 1.1; the volume ratio of HCl to THF was 0.2. In the preparation method (2) of the series IV of PROTAC compounds, the mass ratio of the 2'-OH,3' -OH protected GS-441524 intermediate, the benzodione piperidine derivative, EDCI and 2DMAP is 1; the volume ratio of HCl to THF was 0.2.

The mass ratio of the 2'-OH,3' -OH protected GS-441524 intermediate, the alkyl acid derivative having a terminal Boc amino group, EDCI, and DMAP in the production method (1) of the series V procac compound is 1.1; the volume ratio of HCl to EA is 0.2; the mass ratio of GS-441524 derivative, boc lysine, HATU and DIPEA was 1.1. In the preparation method (2) of the series V of PROTAC compounds, the mass ratio of the 2'-OH,3' -OH protected GS-441524 intermediate, the 9-aminofluorene derivative, EDCI and DMAP is 1.1; the volume ratio of HCl to THF was 0.2. In the preparation method (3) of the PROTAC compound of the series V, the mass ratio of the 2'-OH,3' -OH protected GS-441524 intermediate, the RNF-114 ligand derivative, EDCI and DMAP is 1; the volume ratio of HCl to THF was 0.2.

The ProTAC compound of the Ruidesacvir or the GS-441524 can effectively degrade RNA-dependent RNA polymerase of the EV71 virus, thereby exerting the activity of inhibiting the replication of the EV71 virus, having low toxicity to cells, and being used for preparing the anti-EV 714 virus medicines.

Drawings

FIG. 1 is a Western Blot of a portion of the compounds of the invention on EV71 RdRp degradation.

Detailed Description

Further features and advantages of the present invention will be understood from the following detailed description. The examples provided are merely illustrative of the method of the present invention and do not limit the remainder of the disclosure in any way.

[ example 1 ] preparation of VHL derivatives terminated with Carboxylic acids

The terminal carboxylic acid substituted alkyl chain VHL derivative is synthesized by the following reactions of formula i, ii and iii.

Taking as an example the preparation of the methyl-bearing VHL derivative 6- ((1- ((2s, 4r) -4-hydroxy-2- (((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) carbamoyl) pyrrolidin-1-yl) -3, 3-dimethyl-1-oxobutan-2-yl) amino) hexanoic acid as follows:

(i) 6-Bromohexanoic acid (1.9g, 10.0 mmol) was weighed into a 250mL single-necked flask, and 30mL of tetrahydrofuran was added to dissolve it, after which the system was cooled to-40 ℃. After the system was cooled, trifluoroacetic anhydride (6.2g, 30.0 mmol) was slowly added dropwise thereto, and after stirring at-40 ℃ for 30min, the system was left at room temperature, tert-butanol (6.1g, 80.0 mmol) was added dropwise during the slow warming, and stirring was carried out for 18h. After confirming completion of the reaction by TLC, the organic phase was concentrated, saturated sodium bicarbonate (50 mL) was added thereto, extracted with EA (50 mL. Times.3), and the organic phases were combined and dried over anhydrous sodium sulfate. The organic phase is concentrated and then treated with mobile phase petroleum ether: ethyl acetate =50 (V/V) purification on silica gel column afforded tert-butyl 6-bromohexanoate.

(ii) Tert-butyl 6-bromohexanoate (251.0mg, 1.0mmol) was weighed and placed in a 50mL single-necked flask to be dissolved in acetonitrile (10 mL), and methyl-containing VHL ligand (399.6mg, 0.9mmol), potassium carbonate (414.6mg, 373.2mmol), and potassium iodide (8.3mg, 0.05mmol) were added, followed by placing the reaction at 85 ℃ for overnight reflux. After TLC detection of the reaction was complete, EA (30 mL. Times.3) was extracted with water (30 mL), and the organic phases were combined and dried over anhydrous sodium sulfate. The organic phase was concentrated and washed with mobile phase DCM: meOH =60 (1 (V/V) was purified by silica gel column to give tert-butyl 6- ((1- ((2s, 4r) -4-hydroxy-2- ((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) carbamoyl) pyrrolidin-1-yl) -3, 3-dimethyl-1-oxobutan-2-yl) amino) hexanoate.

(iii) Tert-butyl 6- ((1- ((2S, 4R) -4-hydroxy-2- ((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) carbamoyl) pyrrolidin-1-yl) -3, 3-dimethyl-1-oxobutan-2-yl) amino) hexanoate (300mg, 0.5 mmol) was placed in a 25mL single neck flask, DCM (3 mL) was added thereto, trifluoroacetic acid (3 mL) was added dropwise to the system under ice bath, then the system was warmed to room temperature and stirred for 3h, and the solvent was concentrated after completion of the reaction by TLC monitoring to give 6- ((1- ((2S, 4R) -4-hydroxy-2- (((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) carbamoyl) pyrrolidin-1-yl) -3, 3-dimethyl-1-oxobutan-2-yl) amino) hexanoic acid.

Other terminal carboxylic acid substituted alkyl chain VHL derivatives were prepared as described above.

Example 2 preparation of a VHL derivative terminated with a dialkyl chain of a carboxylic acid

The VHL derivative with the end connected with the double alkyl chain of the carboxylic acid is synthesized by the following reactions of the formulas i, ii and iii.

Taking the preparation of the methyl-bearing VHL derivative 6- ((6- ((1- ((2s, 4r) -4-hydroxy-2- ((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) carbamoyl) pyrrolidin-1-yl) -3, 3-dimethyl-1-oxobutan-2-yl) amino) -6-oxohexyl) amine) hexanoic acid as an example, the procedure was as follows:

(i) VHL (888mg, 2.0mmol) was weighed and placed in a 25mL single-necked flask, DCM (10 mL) was added thereto and dissolved, and then N-Boc-6-aminocaproic acid (569.8mg, 2.2mmol), HATU (1.7g, 4.4mmol), DIPEA (1.0g, 8mmol) were added to the system. The system was then left to stir at room temperature for 3h. After TLC confirmed completion of the reaction, the organic phase was concentrated, saturated sodium bicarbonate (50 mL) was added thereto, extracted with EA (50 mL. Times.3), and the organic phases were combined and dried over anhydrous sodium sulfate. The organic phase was concentrated and washed with mobile phase DCM: meOH =60 (V/V) was purified over silica gel column to give tert-butyl ester of (6- ((1- ((2s, 4r) -4-hydroxy-2- (((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) carbamoyl) pyrrolidin-1-yl) -3, 3-dimethyl-1-oxobutan-2-yl) amino) -6-oxohexyl) carbamate.

(ii) Tert-butyl ester of (6- ((1- ((2s, 4r) -4-hydroxy-2- (((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) carbamoyl) pyrrolidin-1-yl) -3, 3-dimethyl-1-oxobutan-2-yl) amino) -6-oxohexyl) carbamate (657.9mg, 1.0 mmol) was weighed out in a 25mL single vial and dissolved in DCM (10 mL), trifluoroacetic acid (5 mL) was added dropwise to the system under ice bath, then the system was warmed to room temperature and stirred for 3h, tlc monitored for completion of the reaction and the solvent was concentrated to give (2s, 4r) -1- (2- (6-aminocaproamide) -3, 3-dimethylbutyryl) -4-hydroxy-N- ((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) pyrrolidine-2-carboxamide.

(iii) (2S, 4R) -1- (2- (6-aminocaproamide) -3, 3-dimethylbutyryl) -4-hydroxy-N- ((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) pyrrolidine-2-carboxamide (502mg, 0.9 mmol) was placed in a 25mL single vial, to which was added acetonitrile (10 mL). Subsequently, tert-butyl 6-bromohexanoate (251.0mg, 1.0mmol), potassium carbonate (414.6mg, 3mmol), and potassium iodide (8.3mg, 0.05mmol) were added to the system. The reaction was then placed at 85 ℃ under reflux overnight. After TLC detection of the reaction was complete, EA (30 mL. Times.3) was extracted with water (30 mL), and the organic phases were combined and dried over anhydrous sodium sulfate. The organic phase was concentrated and washed with mobile phase DCM: meOH =60 (1 (V/V) was purified by silica gel column to give tert-butyl 6- ((6- ((1- ((2s, 4r) -4-hydroxy-2- (((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) carbamoyl) pyrrolidin-1-yl) -3, 3-dimethyl-1-oxobutan-2-yl) amino) -6-oxohexyl) amino) hexanoate.

(iv) Tert-butyl 6- ((6- ((1- ((2s, 4r) -4-hydroxy-2- (((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) carbamoyl) pyrrolidin-1-yl) -3, 3-dimethyl-1-oxobutan-2-yl) amino) -6-oxohexyl) amino) hexanoate (364mg, 0.5 mmol) was placed in a 25mL single neck flask, DCM (3 mL) was added thereto, trifluoroacetic acid (3 mL) was added dropwise to the system under ice bath, then the system was warmed to room temperature and stirred for 3h, tlc monitored for completion of the reaction and the solvent was concentrated to give 6- ((6- ((1- ((2s, 4r) -4-hydroxy-2- ((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) carbamoyl) pyrrolidin-1-yl) -3, 3-dimethyl-1-oxobutan-2-yl) amino) -6-oxohexyl) amine) hexanoic acid.

Other VHL derivatives terminated by a di-alkyl chain of a carboxylic acid were prepared as described above.

[ example 3] preparation of alkyl chain VHL derivative with bromine at the end

The VHL derivative with the end connected with the double alkyl chain of the carboxylic acid is synthesized by the reaction shown in the following formula i.

Taking as an example the preparation of the methyl bearing VHL derivative (2s, 4r) -1- (2- (10-bromodecanamido) -3, 3-dimethylbutyryl) -4-hydroxy-N- ((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) pyrrolidine-2-carboxamide, the procedure was as follows:

VHL (888mg, 2.0mmol) was weighed and placed in a 25mL single-necked flask, DCM (10 mL) was added thereto and dissolved, and then 10-bromodecanoic acid (555.2mg, 2.2mmol), HATU (836.5g, 2.2mmol), DIPEA (1.0g, 8mmol) were added to the system. The system was then left to stir at room temperature for 3h. After TLC confirmed completion of the reaction, the organic phase was concentrated, saturated sodium bicarbonate (50 mL) was added thereto, extracted with EA (50 mL. Times.3), and the organic phases were combined and dried over anhydrous sodium sulfate. The organic phase was concentrated and purified with mobile phase DCM: meOH =60 (1 (V/V) was purified on silica gel column to give (2s, 4r) -1- (2- (10-bromodecanoylamino) -3, 3-dimethylbutyryl) -4-hydroxy-N- ((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) pyrrolidine-2-carboxamide.

Other alkyl chain VHL derivatives with bromine at the end can be prepared by the same method.

Example 4 preparation of alkyl chain and alkoxy chain chimeric VHL derivatives with Carboxylic acids at the termini

An alkyl chain and alkoxy chain chimeric VHL derivative with the terminal of carboxylic acid is synthesized by the following reactions shown in formulas i, ii, iii and iv.

Taking the preparation of 3- ((2S, 4R) -4-hydroxy-2- (((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) carbamoyl) pyrrolidin-1-yl) -2, 2-dimethyl-17-oxo-7, 10, 13-trioxa-4, 16-diazacyclohexane-20-oic acid as an example, the procedure was as follows:

(i) Monoethyl succinate (1.5g, 10.0mmol) was weighed and placed in a 50mL single-necked bottle, and DCM (30 mL) was added to dissolve it, followed by addition of 2- (2- (2-aminoethoxy) ethoxy) ethan-1-ol (2.1g, 11.0mmol), HATU (4.18g, 11mmol), DIPEA (5.7g, 8mmol). The system was then left to stir at room temperature for 3h. After completion of the reaction was confirmed by TLC, EA (30 mL. Times.3) was extracted with water (30 mL), and the organic phases were combined and dried over anhydrous sodium sulfate. The organic phase was concentrated and washed with mobile phase DCM: meOH =80 (1V/V) was purified by silica gel column to give ethyl-1-hydroxy-13-oxo-3, 6, 9-trioxa-12-hexadecanoic acid-16-ethyl ester.

(ii) Ethyl-1-hydroxy-13-oxo-3, 6, 9-trioxa-12-hexadecanoic acid-16-ethyl ester (642.7 mg, 2mmol) was dissolved in DCM (15 mL), p-toluenesulfonic acid (300mg, 1.116mmol), DIPEA (0.58mL, 3.28mmol) and added thereto at 0 ℃ and the system was warmed to room temperature for 12h. After completion of the reaction was confirmed by TLC, EA (30 mL. Times.3) was extracted with water (30 mL), and the organic phases were combined and dried over anhydrous sodium sulfate. The organic phase was concentrated and purified with mobile phase DCM: meOH =80 (1V/V) was purified on silica gel column to give Tos-substituted ethyl-1-hydroxy-13-oxo-3, 6, 9-trioxa-12-hexadecanoic acid-16-ethyl ester.

(iii) The product of the above step (475.5mg, 1mmol) was dissolved in acetonitrile (10 mL). Then, VHL (444.0mg, 1.0mmol), potassium carbonate (414.6mg, 3mmol), and potassium iodide (8.3mg, 0.05mmol) were added to the system. The reaction was then placed at 85 ℃ under reflux overnight. After TLC detection of the reaction was complete, EA (30 mL. Times.3) was extracted with water (30 mL), and the organic phases were combined and dried over anhydrous sodium sulfate. The organic phase was concentrated and washed with mobile phase DCM: meOH =60 (V/V) was purified through silica gel column.

(iv) The product from the above step was dissolved in EA (5 mL) and lithium hydroxide was added under ice bath to pH =11. Then, the temperature is raised to room temperature for reaction for 0.5h. After TLC monitoring the reaction was complete, PH =3 was adjusted, followed by extraction with EA (30 mL × 3) and water (30 mL), and the organic phases were combined and dried over anhydrous sodium sulfate. The product after organic phase concentration.

Other alkyl chain and alkoxy chain chimeric VHL derivatives with carboxylic acid at the end can be prepared by the same method.

[ example 5 ] preparation of an alkyl chain lenalidomide derivative having a carboxylic acid at the terminal

Synthesizing the terminal carboxylic acid substituted alkyl chain pomalidomide or lenalidomide derivative through the reactions shown in the following formulas i, ii and iii.

Taking the example of the preparation of 6- ((2- (2, 6-dioxopiperidin-3-yl) -1-oxoisoindol-4-yl) amino) hexanoic acid, the procedure was as follows:

(i) 6-Bromohexanoic acid (1.9g, 10.0 mmol) was weighed into a 250mL single-necked flask, and 30mL of tetrahydrofuran was added to dissolve it, after which the system was cooled to-40 ℃. After the system was cooled, trifluoroacetic anhydride (6.2g, 30.0 mmol) was slowly added dropwise thereto, after stirring at-40 ℃ for 30min, the system was left at room temperature, tert-butanol (6.1g, 80.0 mmol) was added dropwise during the slow temperature rise, and stirring was carried out for 18h. After confirming completion of the reaction by TLC, the organic phase was concentrated, saturated sodium bicarbonate (50 mL) was added thereto, extracted with EA (50 mL. Times.3), and the organic phases were combined and dried over anhydrous sodium sulfate. Concentrating the organic phase, and adding mobile phase petroleum ether: ethyl acetate =50 (V/V) purification on silica gel column afforded tert-butyl 6-bromohexanoate.

(ii) Tert-butyl 6-bromohexanoate (348.8mg, 1.39mmol) was dissolved in N-methylpyrrolidone (NMP) (5 mL), and lenalidomide (300mg, 1.16mmol), DIPEA (0.58mL, 3.28mmol) and added thereto, and the system was raised to 110 ℃ for 12 hours. And (3) adding saturated saline solution into the system after the TLC monitoring reaction is completed, fully stirring for 5min, filtering, washing and drying the solid to obtain the tert-butyl 6- ((2- (2, 6-dioxopiperidine-3-yl) -1-oxoisoindol-4-yl) amino) hexanoate.

(iii) Tert-butyl 6- ((2- (2, 6-dioxopiperidin-3-yl) -1-oxoisoindol-4-yl) amino) hexanoate (429.5mg, 1mmol) was dissolved in dichloromethane (5 mL), and trifluoroacetic acid (5 mL) was added under ice bath, followed by warming to room temperature for 3h. Concentration of the organic solvent after TLC monitoring of the reaction was complete afforded 6- ((2- (2, 6-dioxopiperidin-3-yl) -1-oxoisoindol-4-yl) amino) hexanoic acid.

The preparation method of other alkyl chain lenalidomide derivatives with the terminal of carboxylic acid is the same as that of the alkyl chain lenalidomide derivatives.

[ example 6 ] preparation of alkyl benzodione piperidine derivatives having terminal Carboxylic acid

Synthesizing the alkyl chain benzodione piperidine derivative with the substituted terminal carboxylic acid through the reactions shown in the following formulas i, ii, iii and iv.

Taking the preparation of 11- (4- (2, 6-dioxopiperidin-3-yl) phenoxy) undecanoic acid as an example, the procedure is as follows:

(i) 2, 6-bis (benzyloxy) -3-bromopyridine (100mg, 0.27mmol) was weighed and placed in a 25mL single-neck flask, 1, 4-dioxane (3 mL) and water (0.5 mL) were added thereto to dissolve the pyridine, and then p-hydroxyphenylboronic acid (74.5mg, 0.54mmol), potassium phosphate (123mg, 0.58mmol), pdCl were added to the system in this order ₂ (dppf) (22mg, 0.027mmol). The system was replaced with an argon atmosphere and stirred at 110 ℃ for 16h. TLC confirmed the completion of the reaction, and the reaction mixture was filtered through celite, and the resulting filtrate was diluted with EA (10 mL) and washed with saturated sodium chloride water (10 mL. Times.3). The organic phase is dried over anhydrous sodium sulfate, concentrated and then purified with mobile phase petroleum ether: ethyl acetate =3 (1 (V/V) was purified over silica gel column to give 4- (2, 6-bis (benzyloxy) pyridin-3-yl) phenol.

(ii) 4- (2, 6-bis (benzyloxy) pyridin-3-yl) phenol (470mg, 1.2mmol) was dissolved in DMF (10 mL), and then anhydrous potassium carbonate (254.0mg, 1.8mmol), 11-bromoundecanoic acid (432mg, 1.9mmol) were sequentially added thereto, and the system was raised to 85 ℃ to react for 12 hours. TLC confirmed the completion of the reaction, and the reaction mixture was filtered through celite, and the resulting filtrate was diluted with EA (10 mL) and washed with saturated sodium chloride water (10 mL. Times.3). The organic phase is dried over anhydrous sodium sulfate, concentrated and then purified with mobile phase petroleum ether: ethyl acetate =10 (1V/V) to give t-butyl 11- (4- (2, 6-bis (benzyloxy) pyridin-3-yl) phenoxy) undecanoate by silica gel column purification.

(iii) Tert-butyl 4- (4- (2, 6-bis (benzyloxy) pyridin-3-yl) phenoxy) butyrate (450.0mg, 0.8mmol) was dissolved in absolute ethanol (5 mL), and 10% Pd/C (40 mg) was slowly added, followed by replacement of the system with hydrogen and addition of a hydrogen balloon. The system was left to react at room temperature for 12h. TLC monitored the reaction completion, filtered through celite, the filtrate was extracted with EA (10 mL. Times.3), and the organic phases were combined and dried over anhydrous sodium sulfate. The organic phase is concentrated and then treated with mobile phase petroleum ether: ethyl acetate =3 (1V/V) was purified over silica gel column to give tert-butyl 11- (4- (2, 6-dioxopiperidin-3-yl) phenoxy) undecanoate.

(iii) Tert-butyl 4- (4- (2, 6-dioxopiperidin-3-yl) phenoxy) butyrate (300mg, 0.9 mmol) was dissolved in DCM (3 mL), and trifluoroacetic acid (3 mL) was slowly added dropwise under ice bath and stirred at room temperature for 3h. After completion of the TLC monitoring, the reaction was filtered through celite and the organic phase was concentrated to give 11- (4- (2, 6-dioxopiperidin-3-yl) phenoxy) undecanoic acid.

The preparation method of other alkyl-chain benzodione piperidine derivatives with the terminal of carboxylic acid is the same as the above.

[ example 7 ] preparation of alkyl chain 9-aminofluorene derivative having terminal carboxylic acid

Synthesizing the alkyl chain 6- ((9H-fluorene-9-yl) amino) hexanoic acid substituted by the terminal carboxylic acid through the reactions shown in the following formulas i and ii.

(i) The upper 9-aminofluorene (400.0 mg, 2.2mmol) was taken and dissolved in acetonitrile (10 mL). Subsequently, tert-butyl 6-bromohexanoate (607.4mg, 2.4mmol), potassium carbonate (910.8mg, 6.6mmol) and potassium iodide (18.3mg, 0.11mmol) were added to the system. The reaction was then placed at 85 ℃ under reflux overnight. After TLC detection of the reaction was complete, EA (30 mL. Times.3) was extracted with water (30 mL), and the organic phases were combined and dried over anhydrous sodium sulfate. The organic phase is concentrated and then treated with mobile phase petroleum ether: EA =3 (1 (V/V) was purified over silica gel column to give tert-butyl 6- ((9H-fluoren-9-yl) amino) hexanoate.

(ii) Tert-butyl 6- ((9H-fluoren-9-yl) amino) hexanoate (500mg, 1.4 mol) was dissolved in DCM (3 mL), the system was cooled to 0 ℃ and stirred for 5min. Trifluoroacetic acid (3 mL) was slowly added dropwise to the system while cooling on ice, and then the system was warmed to room temperature and stirred for 1h. After the reaction was monitored by TLC, the system was concentrated to give 6- ((9H-fluoren-9-yl) amino) hexanoic acid.

[ example 8 ] preparation of alkyl chain RNF-114 ligand derivative having terminal carboxylic acid

The terminal carboxylic acid substituted alkyl chain 11- (4- (3- (4-bromophenyl) -1- (2-chloroacetyl) -4, 5-dihydro-1H-pyrazol-5-yl) phenoxy) undecanoic acid is synthesized by the reactions shown in the following formulas i, ii, iii and iv.

(i) A mixture of p-hydroxyacetophenone (1.9g, 169mol) and 4-hydroxybenzaldehyde (3.14g, 169mol) in ethanol (5 mL) was added aqueous sodium hydroxide solution (10%, 10 mL), and the reaction was left to react at room temperature for 24 hours. TLC after the reaction was complete, the solution pH =7 was adjusted with hydrochloric acid (10%), and the precipitate was filtered and washed dry with water. The solid was recrystallized from ethanol and water to give pure (E) -1- (4-bromophenyl) -3- (4-hydroxyphenyl) prop-2-en-1-one.

(ii) (E) -1- (4-bromophenyl) -3- (4-hydroxyphenyl) prop-2-en-1-one (200mg, 0.7 mmol) was dissolved in dry DMF, followed by addition of tert-butyl 11-bromoundecanoate (354mg, 1.1mmol), potassium carbonate (366mg, 2.8mmol). The system was replaced with argon and stirred at 60 ℃ for 5h. After TLC confirmed complete consumption of the reaction, the system was allowed to cool to room temperature, and insoluble potassium carbonate was filtered off, followed by extraction with EA (50 mL × 3), organic phases were combined and washed with saturated sodium chloride water, dried over anhydrous sodium sulfate, and the solvent was concentrated in vacuo and then purified by column chromatography (PE: EA = 4) to give tert-butyl (E) -11- (4- (3- (4-bromophenyl) -3-oxopropan-1-yl) phenoxy) undecanoate.

(iii) Tert-butyl (E) -11- (4- (3- (4-bromophenyl) -3-oxopropan-1-yl) phenoxy) undecanoate (200mg, 0.7 mmol) was dissolved in anhydrous ethanol, followed by addition of hydrazine hydrate (175.2mg, 3.5 mmol), replacement of the system with argon, and stirring at 80 ℃ for 5h. After TLC confirmed complete consumption of the reaction, the system was cooled to room temperature, quenched with water and extracted with DCM (10 mL × 3). The organic phases were combined and concentrated to approximately 5mL. Subsequently, chloroethanesulfonyl chloride (117.9mg, 1mmol) and triethylamine (211.1mg, 2.08mmol) were rapidly added thereto under ice bath, and the system was replaced with argon gas and stirred for 0.5h, followed by warming the system to room temperature and stirring overnight. TLC confirmed the reaction was complete and extracted with EA (50 mL × 3), the organic phases were combined and washed with saturated sodium chloride water, dried over anhydrous sodium sulfate, and the solvent was concentrated in vacuo and then purified by column chromatography (PE: EA = 3) to give tert-butyl 11- (4- (3- (4-bromophenyl) -1- (2-chloroacetyl) -4, 5-dihydro-1H-pyrazol-5-yl) phenoxy) undecanoate.

(iv) Tert-butyl 11- (4- (3- (4-bromophenyl) -1- (2-chloroacetyl) -4, 5-dihydro-1H-pyrazol-5-yl) phenoxy) undecanoate (310mg, 0.5 mmol) was dissolved in DCM (3 mL), and trifluoroacetic acid (3 mL) was added thereto under ice-bath, and the system was warmed to room temperature and stirred for 3H. TLC confirmed complete consumption of the reaction and concentrated the solvent in vacuo to afford 11- (4- (3- (4-bromophenyl) -1- (2-chloroacetyl) -4, 5-dihydro-1H-pyrazol-5-yl) phenoxy) undecanoic acid.

Example 9 preparation of ProTAC Compounds of series I Rudexilvir

The ProTAC compound of the series I of the Ruideciclovir is synthesized by the reaction shown in the following formula I.

Taking as an example the preparation of the compound 2-ethylbutyl ((S) - (((2r, 3s,4r, 5r) -5-cyano-3, 4-dihydroxy-5- (4- ((2s, 4r) -4-hydroxy-2- (((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) carbamoyl) pyrrolidin-1-yl) -3, 3-dimethyl-1-oxobutan-2-yl) amino) hexylaminyl) pyrrolo [2,1-f ] [1,2,4] triazin-7-yl) tetrahydrofuran-2-yl) methoxy) (phenoxy) phosphoryl) -L-alanine ester, the procedure is as follows:

reidesciclovir (200mg, 0.33mmol), (S) -7- ((2S, 4R) -4-hydroxy-2- (((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) carbamoyl) pyrrolidin-1-yl) -8, 8-dimethylnonanoic acid (197.4mg, 0.363mmol), HATU (138.0g, 0.363mmol), DIPEA (159mg, 1.32mmol) were weighed. The system was then left to stir at room temperature for 3h. After confirming completion of the reaction by TLC, the organic phase was concentrated, saturated sodium bicarbonate (10 mL) was added thereto, extracted with EA (10 mL. Times.3), and the organic phases were combined and dried over anhydrous sodium sulfate. The organic phase was concentrated and washed with mobile phase DCM: meOH =40 (1 (V/V) was purified over silica gel column to give 2-ethylbutyl ((S) - (((2r, 3s,4r, 5r) -5-cyano-3, 4-dihydroxy-5- (4- ((2s, 4r) -4-hydroxy-2- (((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) carbamoyl) pyrrolidin-1-yl) -3, 3-dimethyl-1-oxobutan-2-yl) amino) hexylaminyl) pyrrolo [2,1-f ] [1,2,4] triazin-7-yl) tetrahydrofuran-2-yl) methoxy) (phenoxy) phosphoryl) -L-alanine ester.

The preparation method of other series I compounds is the same as the above.

[ example 10 ] preparation of PROTAC Compounds of series II GS-441524

The PROTAC compound of the series II GS-441524 is synthesized by the reaction shown in the following formula i.

Taking as an example the preparation of the compound ethyl (2s, 4r) -1- ((S) -2- ((6- ((7- ((2r, 3r,4s, 5r) -2-cyano-3, 4-dihydroxy-5- (hydroxymethyl) tetrahydrofuran-2-yl) pyrrole [2,1-f ] [1,2,4] triazin-4-yl) amino) -6-oxohexyl) amino) -3, 3-dimethylbutyryl) -4-hydroxy-N- ((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) pyrrolidine-2-carboxamide, the procedure is as follows:

GS-441524 (300mg, 1.0mmol), (S) -7- ((2S, 4R) -4-hydroxy-2- (((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) carbamoyl) pyrrolidin-1-yl) -8, 8-dimethylnonanoic acid (543.1mg, 1.1mmol), HATU (418.3mg, 1.1mmol), DIPEA (516.0mg, 4mmol) were weighed. The system was then left to stir at room temperature for 3h. After confirming completion of the reaction by TLC, the organic phase was concentrated, saturated sodium bicarbonate (10 mL) was added thereto, extracted with EA (10 mL. Times.3), and the organic phases were combined and dried over anhydrous sodium sulfate. The organic phase was concentrated and washed with mobile phase DCM: meOH =40 (1 (V/V) column purification on silica gel to obtain the compound ethyl (2s, 4r) -1- ((S) -2- ((6- ((7- ((2r, 3r,4s, 5r) -2-cyano-3, 4-dihydroxy-5- (hydroxymethyl) tetrahydrofuran-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-4-yl) amino) -6-oxohexyl) amino) -3, 3-dimethylbutyryl) -4-hydroxy-N- ((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) pyrrolidine-2-carboxamide by column purification on silica gel.

The preparation method of other series II compounds is the same as that of the compound.

EXAMPLE 11 preparation of PROTAC Compounds of series III (1) GS-441524

PROTAC compounds of series III (1) GS-441524 are synthesized by the reactions shown in the following formulas i and ii.

Taking as an example the preparation of ((2R, 3S,4R, 5R) -5- (4-aminopyrrole [2,1-f ] [1,2,4] triazin-7-yl) -5-cyano-3, 4-dihydroxytetrahydrofuran-2-yl) methyl 6- (((S) -1- ((2S, 4R) -4-hydroxy-2- (((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) carbamoyl) pyrrolidin-1-yl) -3, 3-dimethyl-1-oxobutan-2-yl) amino) hexanoate, the procedure was as follows:

(i) The 2'-OH,3' -OH-protected GS-441524 intermediate (331.1mg, 1.0mmol) was taken and dissolved in DCM (5 mL), and then (S) -7- ((2S, 4R) -4-hydroxy-2- (((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) carbamoyl) pyrrolidin-1-yl) -8, 8-dimethylnonanoic acid (543.1mg, 1.1mmol), EDCI (210.87mg, 1.1mmol), DMAP (146.6mg, 1.2mmol) was added to the system in this order, and reacted at room temperature for 3 hours. TLC confirmed the reaction was complete, and the mixture was extracted with EA (10 mL. Times.3) and water (10 mL), and the organic phases were combined and dried over anhydrous sodium sulfate. The organic phase was concentrated and washed with mobile phase DCM: meOH =40 (MeOH = 40) was purified by silica gel column to obtain a compound ((3ar, 4r,6r, 6ar) -6- (4-aminopyrrole [2,1-f ] [1,2,4] triazin-7-yl) -6-cyano-2, 2-dimethyltetrahydrofuran [3,4-d ] [1,3] dioxin-4-yl) methyl 6- (((S) -1- ((2s, 4r) -4-hydroxy-2- (((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) carbamoyl) pyrrolidin-1-yl) -3, 3-dimethyl-1-oxobutan-2-yl) amino) hexanoate after silica gel column purification.

(ii) The product obtained in the above step was dissolved in THF (1.5 mL), and concentrated hydrochloric acid (0.3 mL) was slowly added dropwise to the system under ice bath, followed by warming to room temperature and stirring for 3h. After completion of the consumption of the reaction as monitored by TLC, saturated sodium bicarbonate (5 mL) was added to the system to pH =7, followed by extraction with EA (10 mL. Times.3), organic phase combination and drying over anhydrous sodium sulfate, followed by concentration to give ((2R, 3S,4R, 5R) -5- (4-aminopyrrole [2,1-f ] [1,2,4] triazin-7-yl) -5-cyano-3, 4-dihydroxytetrahydrofuran-2-yl) methyl 6- (((S) -1- ((2S, 4R) -4-hydroxy-2- (((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) carbamoyl) pyrrolidin-1-yl) -3, 3-dimethyl-1-oxobutan-2-yl) amino) hexanoate

The preparation method of other series III (1) compounds is the same as the above.

[ example 12 ] preparation of PROTAC Compounds of series III (2) GS-441524

PROTAC compounds of series III (2) GS-441524 are synthesized by the reactions shown in the following formulas i and ii.

The procedure used in this series is identical to the preparation of the ProTAC compounds of series III (1) GS-441524 in example 11, differing only in the handedness of the VHL derivative.

EXAMPLE 13 preparation of PROTAC Compounds of series III (3) GS-441524

PROTAC compounds of series III (3) GS-441524 are synthesized by the reaction shown in the following formula i.

Taking as an example the preparation of the compound (2s, 4r) -1- ((S) -2- (10- (((2r, 3s,4r, 5r) -5- (4-aminopyrrole [2,1-f ] [1,2,4] triazin-7-yl) -5-cyano-3, 4-dihydroxytetrahydrofuran-2-yl) methoxy) decanamide) -3, 3-dimethylbutyryl) -4-hydroxy-N- ((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) pyrrolidine-2-carboxamide, the procedure was as follows:

(i) The 2'-OH,3' -OH-protected GS-441524 intermediate (331.1mg, 1.0mmol) was taken and dissolved in acetonitrile (5 mL), followed by addition of (2S, 4S) -1- (2- (10-bromodecanoylamino) -3, 3-dimethylbutyryl) -4-hydroxy-N- ((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) pyrrolidine-2-carboxamide (745.5mg, 1.1mmol), anhydrous potassium carbonate (414mg, 3mmol), potassium iodide (8.3mg, 0.05mmol) in this order. The system was left to react at 85 ℃ for 12h. After TLC confirmed complete consumption of the reaction, the system was allowed to cool to room temperature, EA (20 mL) was added thereto, followed by extraction with EA (20 mL. Times.3), water (20 mL). The organic phases were combined and dried over anhydrous sodium sulfate, concentrated and purified by column chromatography (DCM: meOH =40

(ii) The procedure used was the same as for the preparation of the compound of III (1) in example 11 (ii).

The preparation of the other compounds of series III (3) is as above.

EXAMPLE 14 preparation of PROTAC Compounds of series IV (1) GS-441524

ProTAC compounds of a series IV (1) Rudexilvir and GS-441524 are synthesized by the reactions shown in the following formulas i and ii.

The preparation of this series of compounds is identical to that of the ProTAC compounds of series III (1) GS-441524 from example 11, except that the VHL derivative is replaced by a CRBN derivative.

[ example 15 ] preparation of PROTAC Compounds of series IV (2) GS-441524

ProTAC compounds of a series IV (2) Ruidecivir and GS-441524 are synthesized by the reactions shown in the following formulas i and ii.

The preparation of this series of compounds is identical to that of the PROTAC compounds of series III (1) GS-441524 from example 11, except that the VHL derivative is replaced by a benzodione piperidine derivative.

EXAMPLE 16 preparation of PROTAC Compounds of series V (1) GS-441524

PROTAC compounds of series V (1) GS-441524 were synthesized by reactions shown in the following formulas i, ii, iii.

Taking as an example the preparation of the compound ((2r, 3s,4r, 5r) -5- (4-aminopyrrolo [2,1-f ] [1,2,4] triazin-7-yl) -5-cyano-3, 4-dihydroxytetrahydrofuran-2-yl) 6- ((S) -2, 6-bis ((tert-butoxycarbonyl) amino) hexylamine) hexanoate, the procedure was as follows:

(i) The 2'-OH,3' -OH protected GS-441524 intermediate (331.1mg, 1.0mmol) was dissolved in DCM (15 mL), and then N-Boc-6-aminocaproic acid (254.1mg, 1.1mmol), EDCI (210.87mg, 1.1mmol), DMAP (146.6mg, 1.2mmol) were sequentially added to the system, and reacted at room temperature for 3 hours. TLC confirmed the reaction was complete, and the mixture was extracted with EA (10 mL. Times.3) and water (10 mL), and the organic phases were combined and dried over anhydrous sodium sulfate. The organic phase was concentrated and purified with mobile phase DCM: meOH =80 (1 (V/V) was purified by silica gel column to obtain a compound ((3ar, 4r,6r, 6ar) -6- (4-aminopyrrolo [2,1-f ] [1,2,4] triazin-7-yl) -6-cyano-2, 2-dimethyltetrahydrofuran [3,4-d ] [1,3] dioxolan-4-yl) 6- ((t-butoxycarbonyl) amino) methyl hexanoate after silica gel column purification.

(ii) Methyl ((3aR, 4R,6R, 6aR) -6- (4-aminopyrrolo [2,1-f ] [1,2,4] triazin-7-yl) -6-cyano-2, 2-dimethyltetrahydrofuran [3,4-d ] [1,3] dioxolan-4-yl) 6- ((tert-butoxycarbonyl) amino) hexanoate (544.6 mg, 1mmol) was placed in a 5mL single-neck flask, followed by adding 4mL tetrahydrofuran to the system and placing the system in an ice bath for 5min. After the temperature of the system was lowered to 0 ℃, concentrated hydrochloric acid (0.6 mL) was slowly added dropwise to the system, and then the reaction was left at room temperature for 3 hours. After confirming the completion of the reaction by TLC, the reaction was quenched by adding saturated sodium bicarbonate (5 mL) and extracted with EA (10 mL. Times.3), the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated in vacuo to give methyl ((2R, 3S,4R, 5R) -5- (4-aminopyrrolo [2,1-f ] [1,2,4] triazin-7-yl) -5-cyano-3, 4-dihydroxytetrahydrofuran-2-yl) 6-aminocaproate.

(iii) Methyl ((2R, 3S,4R, 5R) -5- (4-aminopyrrolo [2,1-f ] [1,2,4] triazin-7-yl) -5-cyano-3, 4-dihydroxytetrahydrofuran-2-yl) 6-aminocaproate (300mg, 0.7 mmol) was placed in a 10mL single-neck flask, followed by addition of Boc lysine (266.4mg, 0.77mmol), HATU (292.8 mg,0.8 mmol), DIPEA (361.2mg, 2.8mmol) to the reaction, and the system was left at room temperature for 3h. TLC confirmed the reaction was complete, and the mixture was extracted with EA (10 mL. Times.3) and water (10 mL), and the organic phases were combined and dried over anhydrous sodium sulfate. The organic phase was concentrated and washed with mobile phase DCM: meOH =80 (1 (V/V) purification on silica gel column to give ((2r, 3s,4r, 5r) -5- (4-aminopyrrolo [2,1-f ] [1,2,4] triazin-7-yl) -5-cyano-3, 4-dihydroxytetrahydrofuran-2-yl) 6- ((S) -2, 6-bis ((t-butoxycarbonyl) amino) hexylamino) hexanoate compound by purification on silica gel column.

The ProTAC compounds for the remaining series V (1) GS-441524 were prepared as described above.

EXAMPLE 17 preparation of PROTAC Compounds of series V (2) GS-441524

((2R, 3S,4R, 5R) -5- (4-aminopyrrolo [2,1-f ] [1,2,4] triazin-7-yl) -5-cyano-3, 4-dihydroxytetrahydrofuran-2-yl) 6- ((9H-fluoren-9-yl) amino) hexanoate was synthesized by the reaction shown in the following formulas i and ii.

The preparation of this series of compounds was identical to that of the PROTAC compound of series III (1) GS-441524 in example 11, except that the VHL derivative was replaced with a 9-aminofluorene derivative.

[ example 18 ] PROTAC Compounds of series V (3) GS-441524

((2R, 3S,4R, 5R) -5- (4-aminopyrrolo [2,1-f ] [1,2,4] triazin-7-yl) -5-cyano-3, 4-dihydroxytetrahydrofuran-2-yl) methyl 11- (4- (3- (4-bromophenyl) -1- (2-chloroacetyl) -4, 5-dihydropyrazol-5-yl) phenoxy) undecanoate was synthesized by the reaction shown by the following formulas i and ii.

The preparation of this series of compounds is identical to that of the PROTAC compounds of series III (1) GS-441524 in example 11, except that the VHL derivative is replaced by an RNF-114 ligand derivative.

(1) Compounds numbered R-1 in Table 1

The title compound was prepared in 55% yield as a white solid according to the procedure in example 9.

¹ HNMR(400MHz，Methanol-d ₄ )δ8.89(s，1H)，7.88(s，1H)，7.46–7.40(m，4H)，7.33–7.26(m，2H)，7.20–7.13(m，3H)，6.95–6.89(m，2H)，5.34(dd，J＝5.8，4.0Hz，1H)，5.14(d，J＝5.8Hz，1H)，5.04–4.98(m，1H)，4.71–4.61(m，3H)，4.58–4.54(m，1H)，4.46(s，1H)，4.41–4.32(m，2H)，4.07–4.01(m，1H)，3.97–3.91(m，1H)，3.90–3.83(m，1H)，3.77(d，J＝11.2Hz，1H)，3.71–3.64(m，1H)，3.27–3.22(m，1H)，2.48(s，6H)，2.26–2.20(m，1H)，2.02–1.94(m，1H)，1.77–1.69(m，2H)，1.56–1.43(m，8H)，1.34–1.28(m，9H)，1.04(s，9H)，0.90–0.86(m，6H). ¹³ C NMR(101MHz，Methanol-d ₄ )δ173.03，171.99，155.83，151.47，150.69，147.66，147.04，146.91，144.23，131.96，130.10，129.34，129.19，129.10，129.08，126.24，124.70，119.98，119.94，119.93，119.89，116.84，116.54，116.20，111.37，110.91，101.23，80.43，79.20，74.83，72.96，72.12，70.24，69.55，67.00，66.73，59.04，56.51，50.62，48.75，40.29，37.14，34.74，33.45，29.35，26.30，25.94，24.45，22.84，22.82，21.07，19.10，14.43，9.98，9.93.

(2) Compounds numbered R-2 in Table 1

The title compound was prepared in 59% yield as a white solid by referring to the procedure in example 9.

¹ H NMR(400MHz,Methanol-d ₄ )δ8.89(s,1H)，7.89(d,J＝2.0Hz,1H)，7.44(d,J＝2.5Hz,4H)，7.28(ddd,J＝9.5,5.9,2.2Hz,2H)，7.19–7.11(m,3H)，6.95–6.89(m,2H)，5.37–5.32(m,1H)，5.15(d,J＝5.8Hz,1H)，5.03(t,J＝6.8Hz,2H)，4.71–4.63(m,2H)，4.56(qd,J＝4.0,1.6Hz,1H)，4.46(d,J＝4.7Hz,2H)，4.43–4.32(m,3H)，4.08–4.00(m,1H)，3.98–3.92(m,1H)，1.71(q,J＝7.3Hz,2H)，1.53(dd,J＝12.8,7.1Hz,6H)，1.37–1.30(m,13H). ¹³ C NMR(101MHz,Methanol-d ₄ )δ173.50,173.07,171.94,155.87,151.48,150.62,147.65,147.04,144.22,131.96,130.47,129.34,129.09,126.25,124.71,123.21,119.98,119.52,116.85,116.18,111.43,110.92,101.25,83.19,81.78,80.24,79.18,74.27,72.92,72.16,70.24,69.55,66.92,66.74,65.45,59.05,56.56,54.33,50.08,48.75,40.29,37.18,34.74,33.51,30.78,29.36,28.72,28.56,28.46,26.53,25.94,24.40,22.81,21.08,19.12,14.46,10.00,9.58.

(3) Compounds numbered R-3 in Table 1

The title compound was prepared in 53% yield as a white solid according to the procedure in example 9.

¹ HNMR(400MHz，Methanol-d ₄ )δ8.89(s，1H)，7.87(s，1H)，7.47–7.39(m，4H)，7.33–7.25(m，2H)，7.23–7.12(m，3H)，6.96–6.88(m，2H)，5.35(dd，J＝5.8，4.0Hz，1H)，5.15(d，J＝5.8Hz，1H)，5.03–4.99(d，J＝7.0Hz，1H)，4.70–4.63(m，2H)，4.60–4.52(m，1H)，4.45(s，1H)，4.42–4.39(m，2H)，4.10–4.01(m，1H)，3.98–3.91(m，1H)，3.90–3.81(m，1H)，3.76(d，J＝11.2Hz，1H)，3.67(dd，J＝11.4，3.8Hz，1H)，3.24(s，1H)，2.54–2.44(m，6H)，2.27–2.16(m，1H)，2.03–1.93(m，1H)，1.77–1.66(m，2H)，1.56–1.44(m，6H)，1.41–1.24(m，15H)，1.03(d，J＝6.3Hz，9H)，0.90–0.84(m，6H). ¹³ C NMR(101MHz，Methanol-d ₄ )δ173.53，171.98，157.92，152.16，150.69，147.66，146.92，146.40，143.38，131.96，130.11，129.34，129.09，126.25，124.70，123.22，119.98，119.95，119.90，116.85，116.53，116.15，111.45，110.92，101.24，79.87，79.20，74.27，72.91，72.17，70.25，69.53，67.04，66.75，59.02，56.51，50.09，48.74，40.30，37.15，34.72，33.93，33.52，29.34，28.68，28.62，26.72，25.96，24.43，22.82，21.60，19.51，14.45，9.99，9.04.

(4) Compounds numbered R-4 in Table 1

¹ HNMR(400MHz，Methanol-d ₄ )δ8.89(d，J＝2.0Hz，1H)，7.91–7.84(m，1H)，7.47–7.40(m，4H)，7.33–7.25(m，2H)，7.21–7.10(m，3H)，6.95–6.88(m，2H)，5.35(dd，J＝5.8，3.9Hz，1H)，5.14(dd，J＝8.2，5.7Hz，1H)，5.05–4.98(d，J＝7.0Hz，1H)，4.68(d，J＝8.4Hz，1H)，4.57–4.52(m，1H)，4.48–4.43(m，1H)，4.41–4.31(m，2H)，4.08–4.00(m，1H)，3.97–3.92(m，1H)，3.91–3.82(m，1H)，3.76(d，J＝11.3Hz，1H)，3.70–3.65(m，1H)，3.28(s，1H)，2.54–2.46(m，6H)，2.27–2.18(m，1H)，2.03–1.93(m，1H)，1.76–1.67(m，2H)，1.56–1.45(m，6H)，1.37–1.29(m，17H)，1.04(d，J＝3.8Hz，9H)，0.90–0.86(m，6H). ¹³ C NMR(101MHz，Methanol-d ₄ )δ175.29，173.10，170.92，155.50，150.63，148.27，147.03，142.61，131.96，130.11，129.34，129.21，129.10，126.72，124.70，123.72，120.98，119.96，119.91，112.25，102.50，81.77，79.23，72.89，72.18，69.53，67.02，66.74，59.03，56.53，50.08，48.74，40.30，37.19，35.86，33.54，30.78，29.36，28.92，28.86，28.67，26.84，25.95，23.95，22.85，21.49，19.12，14.46，10.00，9.95.

(5) Compounds numbered R-5 in Table 1

The title compound was prepared in 40% yield as a white solid according to the procedure in example 9.

¹ HNMR(400MHz，Methanol-d ₄ )δ8.89(s，1H)，7.87(s，1H)，7.49–7.41(m，4H)，7.33–7.25(m，2H)，7.23–7.12(m，3H)，6.94–6.89(dd，J＝11.7，4.6Hz，2H)，5.35(dd，J＝5.8，3.9Hz，1H)，5.14(d，J＝5.8Hz，1H)，5.03(s，1H)，4.68–4.63(s，2H)，4.57–4.52(m，1H)，4.48–4.45(m，1H)，4.41–4.30(m，2H)，4.07–4.01(m，1H)，3.97–3.92(m，1H)，3.90–3.81(m，1H)，3.76(d，J＝11.3Hz，1H)，3.67(dd，J＝11.2，3.6Hz，1H)，3.25(s，1H)，2.54–2.45(m，6H)，2.26–2.18(m，1H)，2.03–1.94(m，1H)，1.75–1.66(m，2H)，1.59–1.46(m，6H)，1.38–1.28(m，19H)，1.04(s，9H)，0.90–0.85(m，6H). ¹³ C NMR(101MHz，MeOD)δ173.89，173.01，168.65，151.48，147.66，146.74，142.63，131.96，130.02，129.33，129.22，129.10，125.61，124.70，119.94，119.49，115.71，108.50，101.24，80.15，78.61，74.37，72.89，72.17，69.52，67.06，66.74，58.65，56.50，50.08，48.74，39.76，37.16，35.18，33.53，29.35，29.09，28.90，28.72，26.93，25.95，24.47，22.84，22.81，20.66，19.01，18.50，14.43，10.41，9.94.

(6) Compounds numbered R-6 in Table 1

The title compound was prepared in 34% yield as a white solid according to the procedure in example 9.

¹ HNMR(400MHz，Methanol-d ₄ )δ8.90(s，1H)，7.87(s，1H)，7.46–7.43(m，4H)，7.34–7.24(m，2H)，7.24–7.12(m，3H)，6.97–6.86(m，2H)，5.37(dd，J＝5.7，4.1Hz，1H)，5.13(d，J＝5.7Hz，1H)，5.04–4.98(m，1H)，4.65–4.56(m，2H)，4.46(s，1H)，4.40–4.30(m，3H)，4.28–4.14(m，3H)，4.06–4.00(m，1H)，3.96–3.84(m，3H)，3.79–3.65(m，10H)，3.58–3.54(m，2H)，3.40(t，J＝5.4Hz，2H)，2.79–2.75(m，1H)，2.73–2.64(m，3H)，2.50(d，J＝2.4Hz，5H)，2.26–2.18(m，2H)，2.00–1.94(m，1H)，1.52(dd，J＝7.0，2.3Hz，3H)，1.37–1.33(m，4H)，1.27(d，J＝7.1Hz，3H)，1.07–1.04(m，9H)，0.90–0.85(m，6H). ¹³ C NMR(101MHz，Methanol-d ₄ )δ172.92，172.35，171.85，171.22，158.09，152.23，148.79，146.29，144.91，131.96，130.67，129.33，128.73，126.23，124.69，119.92，116.83，110.98，110.91，109.99，109.20，100.82，79.86，78.60，73.83，73.14，72.10，70.43，69.86，69.60，69.07，66.70，63.60，58.49，56.43，50.08，48.74，40.29，38.78，37.40，35.20，30.87，29.35，28.86，27.71，25.82，25.57，22.82，20.19，18.98，15.30，13.24，9.33.

(7) Compounds numbered R-7 in Table 1

The title compound was prepared in 42% yield as a white solid according to the procedure in example 9.

¹ HNMR(400MHz，Methanol-d ₄ )δ8.90(d，J＝1.9Hz，1H)，7.90(s，1H)，7.45–7.43(m，4H)，7.35–7.25(m，2H)，7.19–7.13(m，3H)，6.95–6.88(m，2H)，5.35(dd，J＝5.8，3.9Hz，1H)，5.14(d，J＝5.8Hz，1H)，5.05–4.98(m，1H)，4.67(s，1H)，4.63–4.56(m，2H)，4.46(s，2H)，4.41–4.33(m，3H)，4.23–4.16(m，3H)，4.07–4.01(m，1H)，3.97–3.83(m，4H)，3.79–3.65(m，9H)，3.56–3.52(m，2H)，3.40–3.35(m，2H)，2.51–2.48(m，5H)，2.28–2.21(d，J＝11.9Hz，4H)，2.01–1.92(m，2H)，1.74–1.72(m，2H)，1.68–1.61(m，2H)，1.52(d，J＝5.0Hz，3H)，1.36–1.34(d，J＝3.8Hz，4H)，1.29–1.26(m，3H)，1.05(s，9H)，0.90–0.85(m，6H). ¹³ C NMR(101MHz，Methanol-d ₄ )δ193.78，172.66，171.85，171.19，157.53，155.74，152.57，147.65，146.89，142.94，137.20，131.96，130.09，129.33，129.09，128.37，126.23，124.70，123.23，119.46，111.27，102.51，79.12，72.94，72.19，70.18，70.11，69.86，69.59，69.18，66.70，63.69，59.57，59.19，56.47，49.94，49.32，46.51，40.30，38.99，36.78，35.26，33.14，31.45，29.36，25.58，24.90，23.93，22.83，22.81，22.34，21.04，19.03，14.42，12.74，9.98，9.27，7.83.

(8) Compounds numbered R-8 in Table 1

The title compound was prepared in 45% yield as a white solid according to the procedure in example 9.

¹ HNMR(400MHz，Methanol-d ₄ )δ8.89(s，1H)，7.89(s，1H)，7.46–7.43(m，4H)，7.33–7.24(m，2H)，7.22–7.12(m，3H)，6.92(q，J＝4.7Hz，2H)，5.35(dd，J＝5.8，3.9Hz，1H)，5.13(d，J＝5.8Hz，1H)，5.04–4.95(m，1H)，4.70–4.59(m，2H)，4.55(dd，J＝4.2，1.7Hz，1H)，4.51–4.41(m，2H)，4.40–4.33(m，3H)，4.28–4.15(m，3H)，4.07–4.01(m，1H)，3.99–3.80(m，4H)，3.75–3.64(m，9H)，3.55(t，J＝5.3Hz，2H)，3.40–3.35(m，2H)，2.49(s，5H)，2.22(t，J＝7.3Hz，4H)，2.02–1.92(m，1H)，1.72(t，J＝7.3Hz，2H)，1.67–1.56(m，4H)，1.52(d，J＝7.1Hz，3H)，1.41–1.35(m，6H)，1.28(d，J＝7.1Hz，3H)，1.05(s，9H)，0.88(t，J＝7.4Hz，6H). ¹³ C NMR(101MHz，Methanol-d ₄ )δ175.61，174.20，172.99，172.25，171.23，157.16，154.81，151.50，150.24，148.11，147.07，144.25，131.46，130.09，129.35，128.37，126.86，125.12，123.25，119.95，119.90，117.26，115.28，112.38，101.25，82.62，79.11，72.97，72.18，70.11，70.06，69.85，69.60，69.34，68.54，66.73，63.48，59.62，59.21，56.59，50.08，49.35，40.29，38.94，37.41，36.01，35.23，33.48，28.43，28.37，25.59，25.43，24.34，23.30，21.84，19.09，19.02，15.48，9.99，9.94.

(9) Compounds numbered R-12 in Table 1

The title compound was prepared in 33% yield as a white solid according to the procedure in example 14.

¹ HNMR(400MHz，Methanol-d ₄ )δ8.88(s，1H)，7.88(d，J＝7.1Hz，1H)，7.44(s，4H)，6.92(q，J＝4.5Hz，2H)，5.09–5.01(m，1H)，4.89(s，1H)，4.68(q，J＝6.5，4.7Hz，1H)，4.52–4.29(m，4H)，4.22–4.14(m，1H)，3.81–3.65(m，2H)，3.25(s，1H)，2.49(s，5H)，2.40–2.17(m，4H)，2.04–1.96(m，1H)，1.54(dd，J＝13.1，6.8Hz，5H)，1.37–1.16(m，12H)，1.04(d，J＝5.2Hz，9H). ¹³ C NMR(101MHz，Methanol-d ₄ )δ173.69，172.00，155.40，151.97，148.07，146.96，144.23，131.97，130.08，126.26，122.09，116.32，102.48，81.99，79.99，74.70，70.67，69.55，66.58，61.92，59.04，56.54，48.76，48.28，36.78，34.73，33.12，29.36，29.07，28.86，28.66，26.93，25.99，24.57，22.57，15.40.

(10) Compounds numbered R-13 in Table 1

The title compound was prepared in 53% yield as a white solid according to the procedure in example 14.

(1) Compounds numbered R-14 in Table 1

The title compound was prepared in 55% yield as a white solid according to the procedure in example 14.

¹ H NMR(400MHz,Chloroform-d)δ7.93(s,1H)，7.34(t,J＝7.8Hz,1H)，7.22(d,J＝7.5Hz,1H)，6.93(dd,J＝4.6,1.3Hz,1H)，6.77(d,J＝2.8Hz,1H)，6.68(d,J＝4.6Hz,1H)，6.37(s,2H)，5.59–5.54(m,1H)，5.49(d,J＝6.7Hz,1H)，5.29–5.05(m,2H)，4.92(dd,J＝6.8,4.2Hz,1H)，4.60(dt,J＝5.9,4.2Hz,1H)，4.51–4.47(m,1H),4.45(dd,J＝12.0,4.1Hz,1H)，4.31–4.27(m,1H)，4.26(d,J＝6.3Hz,1H)，4.15(d,J＝15.6Hz,1H)，3.19(d,J＝7.0Hz,2H)，3.00–2.82(m,3H)，2.31–2.28(m,2H)，2.17–2.12(m,1H)，1.61(dt,J＝15.7,7.6Hz,4H)，1.45–1.36(m,6H)，1.32–1.25(m,12H).

(11) Compounds numbered R-15 in Table 1

The title compound was prepared in 48% yield as a white solid according to the procedure in example 15.

¹ H NMR(400MHz,Methanol-d ₄ )δ7.87(s,1H)，7.33(t,J＝7.8Hz,1H)，7.08(dd,J＝7.6,0.8Hz,1H)，6.91(q,J＝4.6Hz,2H)，6.83(d,J＝8.0Hz,1H)，5.17(dd,J＝13.3,5.2Hz,1H)，4.87(d,J＝5.3Hz,1H)，4.66(s,1H)，4.46–4.30(m,5H)，4.18–4.15(m,1H)，3.21(t,J＝7.1Hz,2H)，2.97–2.88(m,1H)，2.80(ddd,J＝17.8,4.7,2.4Hz,1H)，2.54–2.44(m,1H)，2.31(td,J＝7.4,5.0Hz,2H)，2.19(ddt,J＝10.5,5.4,2.7Hz,1H)，1.65(h,J＝7.4,6.9Hz,2H)，1.60–1.53(m,2H)，1.41(d,J＝7.7Hz,2H)，1.37–1.25(m,14H). ¹³ C NMR(101MHz,Methanol-d ₄ )δ173.72,173.42,171.17,171.00,146.90,143.76,131.49,129.21,126.62,124.21,116.53,116.27,112.42,110.74,110.44,101.22,82.00,80.01,74.25,70.66,62.84,60.18,52.16,48.27,48.05,47.84,47.63,47.42,47.20,46.99,43.05,33.53,31.00,29.22,29.13,29.07,28.87,28.78,28.65,26.81,24.55,22.84,13.09.

(12) Compounds numbered R-16 in Table 1

¹ H NMR(400MHz,Methanol-d ₄ )δ7.88(s,1H)，7.33–7.15(m,2H)，6.94–6.83(m,4H)，4.88(d,J＝5.2Hz,1H)，4.46–4.38(m,2H)，4.33(dd,J＝11.4,4.8Hz,1H)，4.16(t,J＝5.7Hz,1H)，3.93(h,J＝6.5Hz,2H)，2.30(dt,J＝7.4,3.6Hz,2H)，1.78–1.69(m,2H)，1.59–1.54(m,2H)，1.48–1.39(m,2H)，1.36–1.19(m,14H).

(13) Compounds numbered R-17 in Table 1

The title compound was prepared in 39% yield as a white solid according to the procedure of example 9.

¹ HNMR(400MHz，Methanol-d ₄ )δ7.88(s，1H)，7.34–7.29(m，2H)，7.26(d，J＝7.6Hz，1H)，7.19–7.10(m，3H)，7.07(d，J＝7.5Hz，1H)，6.94–6.87(m，2H)，6.80(d，J＝7.9Hz，1H)，5.46–5.40(m，1H)，5.15(q，J＝5.9Hz，2H)，4.56(d，J＝3.9Hz，1H)，4.48–4.23(m，6H)，4.06–3.97(m，1H)，3.97–3.84(m，3H)，3.30–3.16(m，4H)，3.03–2.86(m，2H)，2.83–2.75(m，2H)，2.51–2.35(m，3H)，2.18–1.97(m，3H)，1.66–1.56(m，4H)，1.47–1.43(m，2H)，1.38–1.32(m，16H)，1.28–1.24(d，J＝7.1Hz，3H)，0.87(t，J＝7.4Hz，6H). ¹³ C NMR(101MHz，Methanol-d ₄ )δ176.46，174.51，172.07，171.64，155.86，152.35，147.03，143.78，131.53，129.76，129.23，127.41，125.28，124.15，123.10，119.98，119.90，116.85，112.94，111.52，110.48，101.62，81.72，79.86，78.93，74.27，72.95，72.55，66.75，65.08，53.86，52.15，50.68，45.98，44.86，43.06，40.71，39.36，32.73，31.01，29.07，28.95，28.79，27.31，25.23，22.79，20.17，11.79，10.87.

(14) Compounds numbered R-18 in Table 1

The title compound was prepared in 39% yield as a white solid according to the procedure in example 11.

¹ HNMR(400MHz，Methanol-d ₄ )δ8.88(s，1H)，7.87(d，J＝10.9Hz，1H)，7.44(d，J＝2.6Hz，4H)，6.92(dd，J＝12.5，4.4Hz，2H)，5.02(q，J＝7.0Hz，2H)，4.67(d，J＝8.2Hz，1H)，4.49–4.44(m，2H)，4.42–4.38(m，1H)，4.33–4.30(m，1H)，4.27(td，J＝5.6，4.4Hz，1H)，4.19(dd，J＝6.2，5.2Hz，1H)，3.90–3.75(m，2H)，3.67(dd，J＝11.3，3.7Hz，1H)，2.49(d，J＝1.8Hz，5H)，2.41(dd，J＝8.7，2.7Hz，1H)，2.36–2.20(m，4H)，2.02–1.94(m，1H)，1.61–1.49(m，7H)，1.30(t，J＝7.6Hz，2H)，1.03(d，J＝4.5Hz，9H). ¹³ C NMR(101MHz,Methanol-d ₄ )δ173.58,172.02,155.81,151.53,147.62,146.97,144.24,131.97,130.08,129.09,126.26,124.24,123.99,116.81,116.55,111.87,110.72,101.33,86.64,82.03,79.94,74.30,70.72,69.56,67.07,61.77,59.06,48.95,37.18,34.76,32.60,29.08,26.38,25.99,24.38,21.08,14.48.

(15) Compounds numbered R-19 in Table 1

The title compound was prepared in 25% yield as a white solid according to the procedure in example 11.

¹ HNMR(400MHz，Methanol-d ₄ )δ8.87(d，J＝1.4Hz，1H)，7.88(d，J＝7.8Hz，1H)，7.42(s，4H)，6.96–6.93(m，1H)，6.91(d，J＝4.6Hz，1H)，5.06–4.99(m，2H)，4.70(q，J＝3.6Hz，1H)，4.50–4.40(m，3H)，4.33(dd，J＝7.5，3.9Hz，1H)，4.29(d，J＝4.4Hz，1H)，4.24–4.17(m，1H)，4.10(q，J＝7.2Hz，1H)，3.92–3.73(m，2H)，3.68(dd，J＝11.1，3.5Hz，1H)，3.31(s，1H)，2.48(d，J＝2.0Hz，6H)，2.27(dt，J＝15.5，7.8Hz，3H)，2.21–2.13(m，1H)，1.99(dt，J＝13.5，4.4Hz，1H)，1.52(t，J＝6.7Hz，7H)，1.35–1.26(m，6H)，1.04(d，J＝3.1Hz，9H). ¹³ C NMR(101MHz,Methanol-d ₄ )δ173.70,172.00,155.80,151.54,147.61,147.01,144.19,131.96,130.08,129.09,126.30,124.20,123.94,116.84,116.57,111.93,110.76,101.39,86.73,82.18,79.85,74.34,70.74,69.57,67.10,62.77,60.23,59.10,56.59,48.63,37.22,34.76,33.54,29.30,28.75,26.72,26.05,24.50,21.13,14.58,13.17.

(16) Compounds numbered R-20 in Table 1

The title compound was prepared in 29% yield as a white solid according to the procedure in example 11.

¹ HNMR(400MHz，DMSO-d ₆ )δ9.00(s，1H)，8.42(d，J＝7.6Hz，1H)，7.94(d，J＝1.8Hz，3H)，7.45(d，J＝8.3Hz，2H)，7.38(d，J＝8.3Hz，2H)，6.97–6.76(m，2H)，4.93(d，J＝7.3Hz，1H)，4.67(dd，J＝16.4，4.9Hz，1H)，4.53(t，J＝8.1Hz，1H)，4.35–4.14(m，3H)，4.08–4.01(m，1H)，3.99–3.93(m，1H)，3.69–3.61(m，1H)，3.60–3.48(m，4H)，3.01(s，1H)，2.47(s，3H)，2.42–2.37(m，1H)，2.28(dt，J＝7.5，4.5Hz，2H)，2.19(s，1H)，1.79(ddd，J＝14.9，7.6，3.3Hz，1H)，1.49(t，J＝6.9Hz，2H)，1.40(dd，J＝12.8，6.9Hz，5H)，1.24(d，J＝11.4Hz，12H)，0.92(s，9H).

(17) Compounds numbered R-21 in Table 1

The title compound was prepared in 34% yield as a white solid according to the procedure in example 11.

¹ HNMR(400MHz，Methanol-d ₄ )δ7.88(s，1H)，7.33–7.15(m，2H)，6.94–6.83(m，4H)，4.88(d，J＝5.2Hz，1H)，4.46–4.38(m，2H)，4.33(dd，J＝11.4，4.8Hz，1H)，4.16(t，J＝5.7Hz，1H)，3.93(h，J＝6.5Hz，2H)，2.30(dt，J＝7.4，3.6Hz，2H)，1.78–1.69(m，2H)，1.59–1.54(m，2H)，1.48–1.39(m，2H)，1.36–1.19(m，14H). ¹³ C NMR(101MHz,Methanol-d ₄ )δ173.67,171.69,155.79,151.54,147.63,146.98,144.21,131.96,130.11,129.11,126.28,124.25,124.04,116.82,116.57,111.92,110.81,101.34,86.63,82.06,79.97,74.35,70.77,69.54,66.63,63.55,60.22,59.23,56.77,48.78,48.76,37.38,36.51,34.80,33.59,29.09,28.98,26.64,25.89,25.83,25.78,24.57,21.16,14.60,13.19.

(18) Compounds numbered R-24 in Table 1

¹ H NMR(400MHz,Methanol-d ₄ )δ8.90(s,1H)，7.89(s,1H)，7.47–7.42(m,4H)，6.94–6.89(m,2H)，5.03(t,J＝7.0Hz,2H)，4.86(s,1H)，4.69(dd,J＝9.1,7.8Hz,1H)，4.50–4.29(m,5H)，4.17–4.13(m,1H)，3.79(d,J＝11.2Hz,1H)，3.67(dd,J＝11.1,3.6Hz,1H)，3.41(d,J＝3.8Hz,1H)，2.56(tt,J＝9.2,4.2Hz,2H)，2.50(s,3H)，2.32(td,J＝7.4,4.7Hz,2H)，2.27–2.17(m,2H)，1.99(ddd,J＝13.4,9.1,4.4Hz,1H)，1.55(dd,J＝11.5,7.5Hz,7H)，1.36–1.27(m,14H)，1.06(d,J＝3.9Hz,9H).

(19) Compounds numbered R-25 in Table 1

The title compound was prepared in 34% yield as a white solid according to the procedure in example 13.

¹ H NMR(400MHz,Chloroform-d)δ8.71(s,1H)，7.94(s,1H)，7.49(dd,J＝9.4,7.4Hz,1H)，7.47–7.37(m,4H)，7.00(d,J＝4.6Hz,1H)，6.68(d,J＝4.6Hz,1H)，6.52–6.40(m,2H)，5.44(d,J＝6.6Hz,1H)，5.31(s,1H)，5.28–5.23(m,1H)，5.14–5.08(m,1H)，4.73–4.66(m,2H)，4.60(d,J＝8.9Hz,1H)，4.51(s,1H)，4.08(d,J＝11.3Hz,1H)，3.98(dd,J＝12.5,1.9Hz,1H)，3.85(d,J＝12.3Hz,1H)，3.64(dd,J＝11.4,3.8Hz,1H)，3.56(dq,J＝13.5,6.8Hz,2H)，2.53(s,3H)，2.44–2.34(m,1H)，2.16(t,J＝7.5Hz,2H)，1.80(s,3H)，1.61(dd,J＝19.3,12.0Hz,4H)，1.51(d,J＝6.9Hz,3H)，1.40(s,3H)，1.27(dd,J＝8.5,5.8Hz,12H)，1.05(s,9H)。

(20) Compounds numbered R-27 in Table 1

The title compound was prepared in 45% yield as a white solid according to the procedure in example 11.

¹ H NMR(400MHz,Methanol-d ₄ )δ8.91(s,1H)，8.18–8.07(m,1H)，7.86(s,1H)，7.50–7.40(m,4H)，7.01–6.93(m,2H)，5.05(s,2H)，4.74–4.60(m,4H)，4.48(s,1H)，4.33–4.29(m,1H)，4.25(dd,J＝5.5,4.3Hz,1H)，3.94–3.80(m,3H)，3.76(dd,J＝12.4,3.8Hz,1H)，3.66–3.62(m,1H)，3.26(s,1H)，3.21(dd,J＝7.2,1.8Hz,2H)，2.82(s,2H)，2.50(s,3H)，2.23(dt,J＝19.7,7.5Hz,4H)，2.07–1.95(m,2H)，1.83–1.77(m,2H)，1.62(s,4H)，1.54(d,J＝7.0Hz,3H)，1.34(d,J＝3.9Hz,14H)，1.15(s,9H).

(21) Compounds numbered R-30 in Table 1

The title compound was prepared in 49% yield as a white solid according to the procedure in example 10.

¹ H NMR(400MHz,DMSO-d ₆ )δ9.00(s,1H)，8.47(d,J＝7.6Hz,1H)，8.04(s,1H)，7.94(d,J＝5.3Hz,1H)，7.48–7.35(m,4H)，6.97–6.88(m,2H)，6.51(d,J＝6.3Hz,1H)，5.20(dd,J＝5.7,3.7Hz,1H)，5.11(t,J＝5.4Hz,2H)，5.01–4.96(m,1H)，4.92(t,J＝7.2Hz,1H)，4.54(t,J＝8.2Hz,1H)，4.32–4.22(m,2H)，3.67–3.53(m,3H)，3.50(d,J＝4.2Hz,1H)，3.18(d,J＝3.1Hz,1H)，2.47(s,3H)，2.39(t,J＝7.3Hz,2H)，2.07(t,J＝10.7Hz,1H)，1.82–1.73(m,1H)，1.66–1.54(m,2H)，1.44–1.35(m,5H)，1.35–1.18(m,12H)，0.95(s,9H).

(22) Compounds numbered R-31 in Table 1

The title compound was prepared in 57% yield as a white solid according to the procedure in example 10.

¹ H NMR(400MHz,DMSO-d ₆ )δ9.00(s,1H)，8.47(s,1H)，7.93(s,2H)，7.47–7.37(m,4H)，6.92(d,J＝4.5Hz,1H)，6.90(s,1H)，5.24(d,J＝5.2Hz,1H)，4.96(s,1H)，4.66(s,1H)，4.55(t,J＝8.3Hz,1H)，4.30(s,1H)，4.15(q,J＝5.2Hz,1H)，4.08(q,J＝4.4Hz,2H)，3.97(q,J＝5.3Hz,2H)，3.69–3.58(m,3H)，3.55–3.50(m,2H)，3.19(s,1H)，2.47(s,3H)，2.27(t,J＝7.3Hz,2H)，2.10–2.02(m,1H)，1.84–1.74(m,1H)，1.50–1.45(m,2H)，1.43–1.35(m,5H)，1.32–1.19(m,4H)，0.96(s,9H).

(23) Compounds numbered R-32 in Table 1

The title compound was prepared in 55% yield as a white solid according to the procedure in example 10.

¹ H NMR(400MHz,Methanol-d ₄ )δ8.89(s,1H)，7.85(d,J＝1.9Hz,1H)，7.47–7.41(m,4H)，7.03–6.95(m,1H)，6.96–6.90(m,1H)，5.40(dd,J＝5.9,2.8Hz,1H)，5.20(dd,J＝5.9,3.4Hz,1H)，5.07–5.00(m,1H)，4.67(t,J＝8.4Hz,2H)，4.50–4.40(m,2H)，3.92–3.61(m,5H)，3.37(s,1H)，3.32(s,2H)，2.56–2.45(m,7H)，2.35–2.18(m,2H)，2.05–2.14(m,1H)，1.73(t,J＝7.2Hz,2H)，1.58–1.47(m,5H)，1.40–1.26(m,14H)，1.04(s,9H).

(24) Compounds numbered R-36 in Table 1

The title compound was prepared in 55% yield as a white solid by referring to the procedure in example 18.

¹ H NMR(400MHz,Methanol-d ₄ )δ7.87(d,J＝2.5Hz,1H)，7.77–7.73(m,2H)，7.66–7.61(m,2H)，7.19–7.13(m,2H)，6.91–6.85(m,4H)，5.56(dd,J＝11.7,4.6Hz,1H)，5.51(s,1H)，4.87(dd,J＝5.3,2.2Hz,1H)，4.46–4.29(m,4H)，4.16–4.13(m,1H)，3.93(t,J＝6.4Hz,2H)，3.37(s,1H)，3.21(dd,J＝18.2,4.5Hz,1H)，2.33–2.28(m,2H)，1.73(t,J＝7.4Hz,2H)，1.58–1.54(m,2H)，1.42(d,J＝8.7Hz,2H)，1.31(d,J＝3.7Hz,12H).

(25) Compounds numbered R-37 in Table 1

¹ H NMR(400MHz,DMSO-d ₆ )δ9.00(s,1H)，8.42(d,J＝7.6Hz,1H)，7.94(d,J＝1.8Hz,3H)，7.45(d,J＝8.3Hz,2H)，7.38(d,J＝8.3Hz,2H)，6.97–6.76(m,2H)，4.93(d,J＝7.3Hz,1H)，4.67(dd,J＝16.4,4.9Hz,1H)，4.53(t,J＝8.1Hz,1H)，4.35–4.14(m,3H)，4.08–4.01(m,1H)，3.99–3.93(m,1H)，3.69–3.61(m,1H)，3.60–3.48(m,4H)，3.01(s,1H)，2.47(s,3H)，2.42–2.37(m,1H)，2.28(dt,J＝7.5,4.5Hz,2H)，2.19(s,1H)，1.79(ddd,J＝14.9,7.6,3.3Hz,1H)，1.49(t,J＝6.9Hz,2H)，1.40(dd,J＝12.8,6.9Hz,5H)，1.24(d,J＝11.4Hz,12H)，0.92(s,9H).

The chemical structures of the target compounds of the present invention synthesized above are shown in table 1. The other compounds in table 1 were prepared by the methods described above.

[ example 20 ] ProTAC Compound bioactivity assay of Rudexilvir or GS-441524

(1) ProTAC compounds of Reidesciclovir or GS-441524 in vitro anti-EV 71 virus activity:

RD cells were plated at 3X 10 per well ⁴ Density of individual cells seeded in 96-well plates at 37 ℃ and 5% ₂ And culturing for 24 hours. Cells were infected with EV71 virus with multiplicity of infection (MOI) of 1 and adsorbed for 1h. RD cells were then treated with 1 μ L of different starting concentrations of test compound per column, with 10-fold gradient dilution in each well. After 24h and 48h of infection, the growth status of the cells in each well was observed by microscope, the cells administered at different concentrations were compared with normal non-administered cells, the change in cell status or the death rate was marked, and the cytopathic effect (CPE) was measured.

(2) ProTAC compound cytotoxicity assays of Rudexilvir or GS-441524:

100 μ L of 1X 10 in 96-well plates ⁵ Cell suspension per well, plates at 37 ℃ and 5% CO in incubator ₂ Preculture for 24h. To the plates 10. Mu.L of different concentrations of the test compound were added and the plates were incubated in an incubator for 24h. Add 10. Mu.L of CCK solution to each well and incubate the plates in the incubator for 1-4h. Absorbance at 450nm was measured with a microplate reader. Cell viability was analyzed and graphs generated using software graphpadprism 8.0.

(3) ProTAC compound RdRp degrading activity of Rudexilvir or GS-441524:

RD cells were plated in 6-well plates, the medium was changed to serum-free medium when the cells had grown to a density of about 60%, 2. Mu.g/well of a plasmid expressing EV71 virus RNA-dependent RNA polymerase protein was transfected with PEi (polyethyleneimine) at a mass-to-volume ratio of 1 ₂ After 24 hours of incubation under the conditions, the supernatant was discarded, the cells were collected, and the content of RdRp protein was measured by the method of Western Blot after RAPI cleavage by adding a lysis solution containing 1% of PMSF (phenylmethylsulfonyl chloride) and 10% of cocktail.

The invention takes the Reidesciclovir and GS-441524 as contrast, carries out cytotoxicity and virus inhibition activity and protein degradation activity on the synthesized compound, and all activity results are calculated by mu M.

TABLE 2 in vitro anti-EV 71 results of a portion of the compounds synthesized according to the invention

As can be seen from the table, most compounds have good inhibitory activity on EV71 virus, and the compounds R13, R14, R20, R21, R24 and R25 are obviously superior to the parent molecule GS-441524 of the control medicament and are equivalent to the Reidesvir. On the other hand, R13, R14, R20, R21, R24 and R25 can be degraded by a key enzyme RdRp required by EV71 replication in a targeted way in vitro, so that the activity of the compound is obviously improved compared with the active metabolic component GS-441524 of the Reidesvir, and the effect equivalent to the Reidesvir is achieved.

Since PROTACs can be used to overcome drug resistance, it is an attractive strategy to be able to apply the PROTAC technology to the field of antiviral. Since the RdRp enzyme is a key enzyme required for the replication of most RNA viruses and has significant homology at the catalytic center, this makes PROTACs capable of degrading the RdRp enzyme required for viral replication, in turn, the most promising direction among antiviral PROTACs. PROTACs do not need to have high affinity to target proteins when inducing protein degradation, and the compounds based on Reidesciclovir or GS-441524 are expected to overcome gradual Reidesciclovir resistant strains, are EV71 RdRp inhibitors with good activity and very promising prospect, and can be used for preparing anti-EV 71 medicaments.

The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.

Claims

1. A ProTAC compound of Reidesciclovir or GS-441524, characterized in that: the structure of the ProTAC compound of the Redcixvir is shown as a general formula (I); the structure of the PROTAC compound of GS-441524 is shown as a general formula (II) or (III);

in the general formula, the E3ligase ligand is a protease micromolecule ligand with ubiquitination function and coded by VHL, CRBN or RNF114 genes, or a hydrophobic label micromolecule ligand; linker is-alkylene or-alkoxy.

2. ProTAC compounds of Rudexilvir or GS-441524 according to claim 1, characterized in that: the E3 ligand is selected from benzodiazepine piperidine, lenalidomide, methyllenalidomide, methyl-bearing VHL, trans-bearing methyl VHL, RNF-114, boc lysine, and 9-aminofluorene.

3. ProTAC compounds of Rudexilvir or GS-441524 according to claim 1, characterized in that: the alkylene or alkoxy radical being optionally substituted by oneOr a linear or branched alkylene or alkoxy radical interrupted once or more than once by one of the following groups: - (CH) ₂ ) _n -、-(CH ₂ ) _n CO-、-NR ₁ (CH ₂ ) _n CO-、-NR ₂ (CH ₂ ) _n -、-(OCH ₂ CH ₂ O) _n -、-(CH ₂ CH ₂ O) _n -、-(OCH ₂ CH ₂ OCH ₂ ) _n -、-(CH ₂ CH ₂ OCH ₂ ) _n -、-(CH ₂ CH ₂ OCH ₂ CH ₂ ) _n -, heteroarylene group or any combination thereof, wherein n represents a natural number from 1 to 20, R ₁ 、R ₂ Each independently of the other being H or C _1-10 An alkyl group.

4. ProTAC compounds of Rudexilvir or GS-441524 according to claim 1, characterized in that: selected from the following compounds: 2-ethylbutyl ((S) - (((2r, 3s,4r, 5r) -5-cyano-3, 4-dihydroxy-5- (4- ((2s, 4r) -4-hydroxy-2- (((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) carbamoyl) pyrrolidin-1-yl) -3, 3-dimethyl-1-oxobutan-2-yl) amino) hexylaminyl) pyrrolo [2,1-f ] [1,2,4] triazin-7-yl) tetrahydrofuran-2-yl) methoxy) (phenoxy) phosphoryl) -L-alanine ester;

2-ethylbutyl ((S) - (((2r, 3s,4r, 5r) -5-cyano-3, 4-dihydroxy-5- (4- ((2s, 4r) -4-hydroxy-2- (((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) carbamoyl) pyrrolidin-1-yl) -3, 3-dimethyl-1-oxobutan-2-yl) amino) heptylamino) pyrrolo [2,1-f ] [1,2,4] triazin-7-yl) tetrahydrofuran-2-yl) methoxy) (phenoxy) phosphoryl) -L-alanine ester;

2-ethylbutyl ((S) - (((2r, 3s,4r, 5r) -5-cyano-3, 4-dihydroxy-5- (4- ((2s, 4r) -4-hydroxy-2- (((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) carbamoyl) pyrrolidin-1-yl) -3, 3-dimethyl-1-oxobutan-2-yl) amino) octanamide) pyrrolo [2,1-f ] [1,2,4] triazin-7-yl) tetrahydrofuran-2-yl) methoxy) (phenoxy) phosphoryl) -L-alanine ester;

2-ethylbutyl ((S) - (((2r, 3s,4r, 5r) -5-cyano-3, 4-dihydroxy-5- (4- ((2s, 4r) -4-hydroxy-2- (((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) carbamoyl) pyrrolidin-1-yl) -3, 3-dimethyl-1-oxobutan-2-yl) amino) nonyl) pyrrolo [2,1-f ] [1,2,4] triazin-7-yl) tetrahydrofuran-2-yl) methoxy) (phenoxy) phosphoryl) -L-alanine ester;

2-ethylbutyl ((S) - ((2r, 3s,4r, 5r) -5-cyano-3, 4-dihydroxy-5- (4- ((2s, 4r) -4-hydroxy-2- (((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) carbamoyl) pyrrolidin-1-yl) -3, 3-dimethyl-1-oxobutan-2-yl) amino) decanamide) pyrrolo [2,1-f ] [1,2,4] triazin-7-yl) tetrahydrofuran-2-yl) methoxy) (phenoxy) phosphoryl) -L-alanine ester;

2-ethylbutyl ((S) - ((2r, 3s,4r, 5r) -5-cyano-3, 4-dihydroxy-5- (4- ((2s, 4r) -4-hydroxy-2- (((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) carbamoyl) pyrrolidine-1-carbonyl) -2, 2-dimethyl-17-oxo-7, 10, 13-trioxa-4, 16-diazaspiro-20-amino) pyrrolo [2,1-f ] [1,2,4] triazin-7-yl) tetrahydrofuran-2-yl) methoxy) (phenoxy) phosphoryl) -L-alanine ester;

2-ethylbutyl ((S) - (((2r, 3s,4r, 5r) -5-cyano-3, 4-dihydroxy-5- (4- ((2s, 4r) -4-hydroxy-2- (((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) carbamoyl) pyrrolidine-1-carbonyl) -2, 2-dimethyl-17-oxo-7, 10, 13-trioxa-4, 16-diazacyclobutane-22-amino) pyrrolo [2,1-f ] [1,2,4] triazin-7-yl) tetrahydrofuran-2-yl) methoxy) (phenoxy) phosphoryl) -L-alanine ester;

2-ethylbutyl ((S) - ((2r, 3s,4r, 5r) -5-cyano-3, 4-dihydroxy-5- (4- ((2s, 4r) -4-hydroxy-2- (((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) carbamoyl) pyrrolidine-1-carbonyl) -2, 2-dimethyl-17-oxo-7, 10, 13-trioxa-4, 16-diazatetrac-24-amino) pyrrolo [2,1-f ] [1,2,4] triazin-7-yl) tetrahydrofuran-2-yl) methoxy) (phenoxy) phosphoryl) -L-alanine ester;

((2R, 3S,4R, 5R) -5- (4-aminopyrrolo [2,1-f ] [1,2,4] triazin-7-yl) -5-cyano-3, 4-dihydroxytetrahydrofuran-2-yl) 6- ((2- (2, 6-dioxopiperidin-3-yl) -1-isoindol-4-yl) amino) hexanoate;

((2R, 3S,4R, 5R) -5- (4-aminopyrrolo [2,1-f ] [1,2,4] triazin-7-yl) -5-cyano-3, 4-dihydroxytetrahydrofuran-2-yl) 8- ((2- (2, 6-dioxopiperidin-3-yl) -1-isoindol-4-yl) amino) octanoate;

((2R, 3S,4R, 5R) -5- (4-aminopyrrolo [2,1-f ] [1,2,4] triazin-7-yl) -5-cyano-3, 4-dihydroxytetrahydrofuran-2-yl) 10- ((2- (2, 6-dioxopiperidin-3-yl) -1-isoindol-4-yl) amino) nonanoate;

((2R, 3S,4R, 5R) -5- (4-aminopyrrolo [2,1-f ] [1,2,4] triazin-7-yl) -5-cyano-3, 4-dihydroxytetrahydrofuran-2-yl) 10- ((2- (2, 6-dioxopiperidin-3-yl) -1-isoindol-4-yl) amino) decanoate;

((2R, 3S,4R, 5R) -5- (4-aminopyrrolo [2,1-f ] [1,2,4] triazin-7-yl) -5-cyano-3, 4-dihydroxytetrahydrofuran-2-yl) 11- ((2- (2, 6-dioxopiperidin-3-yl) -1-isoindol-4-yl) amino) undecanoate;

((2R, 3S,4R, 5R) -5- (4-aminopyrrolo [2,1-f ] [1,2,4] triazin-7-yl) -5-cyano-3, 4-dihydroxytetrahydrofuran-2-yl) methyl 11- ((2- (1-methyl-2, 6-dioxopiperidin-3-yl) -1-isoindolin-4-yl) amino) undecanoate;

((2R, 3S,4R, 5R) -5- (4-aminopyrrolo [2,1-f ] [1,2,4] triazin-7-yl) -5-cyano-3, 4-dihydroxytetrahydrofuran-2-yl) 12- ((2- (2, 6-dioxopiperidin-3-yl) -1-isoindol-4-yl) amino) dodecanoic acid methyl ester;

((2R, 3S,4R, 5R) -5- (4-aminopyrrolo [2,1-f ] [1,2,4] triazin-7-yl) -5-cyano-3, 4-dihydroxytetrahydrofuran-2-yl) 10- (4- (2, 6-dioxopiperidin-3-yl) phenoxy) decanoate;

2-ethylbutyl ((S) - (((2r, 3s,4r, 5r) -5-cyano-5- (4- (1- (10- (2, 6-dioxopiperidin-3-yl) -1-oxoisoindolin-4-yl) amino) decanoyl) piperidine-4-carboxamide) pyrrolo [2,1-f ] [1,2,4] triazin-7-yl) -3, 4-dihydroxytetrahydrofuran-2-yl) methoxy) (phenoxy) phosphoryl) -L-alanine ester;

((2R, 3S,4R, 5R) -5- (4-aminopyrrole [2,1-f ] [1,2,4] triazin-7-yl) -5-cyano-3, 4-dihydroxytetrahydrofuran-2-yl) methyl 6- (((S) -1- ((2S, 4R) -4-hydroxy-2- (((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) carbamoyl) pyrrolidin-1-yl) -3, 3-dimethyl-1-oxobutan-2-yl) amino) hexanoate;

((2R, 3S,4R, 5R) -5- (4-aminopyrrole [2,1-f ] [1,2,4] triazin-7-yl) -5-cyano-3, 4-dihydroxytetrahydrofuran-2-yl) methyl 8- (((S) -1- ((2S, 4R) -4-hydroxy-2- (((R) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) carbamoyl) pyrrolidin-1-yl) -3, 3-dimethyl-1-oxobutan-2-yl) amino) octanoate;

((2R, 3S,4R, 5R) -5- (4-aminopyrrole [2,1-f ] [1,2,4] triazin-7-yl) -5-cyano-3, 4-dihydroxytetrahydrofuran-2-yl) methyl 10- (((S) -1- ((2S, 4R) -4-hydroxy-2- (((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) carbamoyl) pyrrolidin-1-yl) -3, 3-dimethyl-1-oxobutan-2-yl) amino) decanoate;

((2R, 3S,4R, 5R) -5- (4-aminopyrrole [2,1-f ] [1,2,4] triazin-7-yl) -5-cyano-3, 4-dihydroxytetrahydrofuran-2-yl) methyl 11- (((S) -1- ((2S, 4R) -4-hydroxy-2- (((R) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) carbamoyl) pyrrolidin-1-yl) -3, 3-dimethyl-1-oxobutan-2-yl) amino) undecanoate;

((2R, 3S,4R, 5R) -5- (4-aminopyrrole [2,1-f ] [1,2,4] triazin-7-yl) -5-cyano-3, 4-dihydroxytetrahydrofuran-2-yl) methyl 10- (((S) -1- ((2R, 4R) -4-hydroxy-2- (((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) carbamoyl) pyrrolidin-1-yl) -3, 3-dimethyl-1-oxobutan-2-yl) amino) decanoate;

((2R, 3S,4R, 5R) -5- (4-aminopyrrole [2,1-f ] [1,2,4] triazin-7-yl) -5-cyano-3, 4-dihydroxytetrahydrofuran-2-yl) methyl 11- (((S) -1- ((2R, 4R) -4-hydroxy-2- (((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) carbamoyl) pyrrolidin-1-yl) -3, 3-dimethyl-1-oxobutan-2-yl) amino) decanoate;

(2S, 4R) -1- ((S) -2- (10- (((2R, 3S,4R, 5R) -5- (4-aminopyrrole [2,1-f ] [1,2,4] triazin-7-yl) -5-cyano-3, 4-dihydroxytetrahydrofuran-2-yl) methoxy) decanamide) -3, 3-dimethylbutyryl) -4-hydroxy-N- ((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) pyrrolidine-2-carboxamide;

(2S, 4R) -1- ((S) -2- (11- (((2R, 3S,4R, 5R) -5- (4-aminopyrrole [2,1-f ] [1,2,4] triazin-7-yl) -5-cyano-3, 4-dihydroxytetrahydrofuran-2-yl) methoxy) decanamide) -3, 3-dimethylbutyryl) -4-hydroxy-N- ((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) pyrrolidine-2-carboxamide;

((2R, 3S,4R, 5R) -5- (4-aminopyrrole [2,1-f ] [1,2,4] triazin-7-yl) -5-cyano-3, 4-dihydroxytetrahydrofuran-2-yl) methyl 4- ((8- (((S) -1- ((2S, 4R) -4-hydroxy-2- (((R) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) carbamoyl) pyrrolidin-1-yl) -3, 3-dimethyl-1-oxobutan-2-yl) amino) butanoate;

((2R, 3S,4R, 5R) -5- (4-aminopyrrole [2,1-f ] [1,2,4] triazin-7-yl) -5-cyano-3, 4-dihydroxytetrahydrofuran-2-yl) methyl 4- ((10- (((S) -1- ((2S, 4R) -4-hydroxy-2- (((R) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) carbamoyl) pyrrolidin-1-yl) -3, 3-dimethyl-1-oxobutan-2-yl) amino) -10-oxodecyl) butanoate;

((2R, 3S,4R, 5R) -5- (4-aminopyrrole [2,1-f ] [1,2,4] triazin-7-yl) -5-cyano-3, 4-dihydroxytetrahydrofuran-2-yl) methyl 6- ((10- (((S) -1- ((2S, 4R) -4-hydroxy-2- (((R) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) carbamoyl) pyrrolidin-1-yl) -3, 3-dimethyl-1-oxobutan-2-yl) amino) -10-oxodecyl) hexanoic acid ester;

((2r, 3s,4r, 5r) -5- (4-aminopyrrole [2,1-f ] [1,2,4] triazin-7-yl) -5-cyano-3, 4-dihydroxytetrahydrofuran-2-yl) methyl (S) -3- ((2r, 4r) -4-hydroxy-2- (((R) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) carbamoyl) pyrrolidin-1-yl) -2, 2-dimethyl-17-oxo-7, 10, 13-trioxa-4, 16-diazetide-20-carboxylate;

(2s, 4r) -1- ((S) -2- ((6- ((7- ((2r, 3s,4r, 5r) -2-cyano-3, 4-dihydroxy-5- (hydroxymethyl) tetrahydrofuran-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-4-yl) amino) -6-oxohexyl) amino) -3, 3-dimethylbutyryl) -4-hydroxy-N- ((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) pyrrolidine-2-carboxamide;

(2s, 4r) -1- ((S) -2- ((6- ((7- ((2r, 3s,4r, 5r) -2-cyano-3, 4-dihydroxy-5- (hydroxymethyl) tetrahydrofuran-2-yl) pyrrole [2,1-f ] [1,2,4] triazin-4-yl) amino) -8-oxooctyl) amino) -3, 3-dimethylbutyryl) -4-hydroxy-N- ((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) pyrrolidine-2-carboxamide;

(2s, 4r) -1- ((S) -2- ((6- ((7- ((2r, 3s,4r, 5r) -2-cyano-3, 4-dihydroxy-5- (hydroxymethyl) tetrahydrofuran-2-yl) pyrrole [2,1-f ] [1,2,4] triazin-4-yl) amino) -10-oxodecyl) amino) -3, 3-dimethylbutyryl) -4-hydroxy-N- ((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) pyrrolidine-2-carboxamide;

((2R, 3S,4R, 5R) -5- (4-aminopyrrolo [2,1-f ] [1,2,4] triazin-7-yl) -5-cyano-3, 4-dihydroxytetrahydrofuran-2-yl) 6- ((S) -2, 6-bis ((t-butoxycarbonyl) amino) hexylamino) hexanoate ester;

((2R, 3S,4R, 5R) -5- (4-aminopyrrolo [2,1-f ] [1,2,4] triazin-7-yl) -5-cyano-3, 4-dihydroxytetrahydrofuran-2-yl) methyl 11- ((S) -2, 6-bis ((t-butoxycarbonyl) amino) hexanamide) undecanoate;

((2R, 3S,4R, 5R) -5- (4-aminopyrrolo [2,1-f ] [1,2,4] triazin-7-yl) -5-cyano-3, 4-dihydroxytetrahydrofuran-2-yl) 6- ((9H-fluoren-9-yl) amino) hexanoate;

((2R, 3S,4R, 5R) -5- (4-aminopyrrolo [2,1-f ] [1,2,4] triazin-7-yl) -5-cyano-3, 4-dihydroxytetrahydrofuran-2-yl) methyl 11- (4- (3- (4-bromophenyl) -1- (2-chloroacetyl) -4, 5-dihydropyrazol-5-yl) phenoxy) undecanoate;

((2R, 3S,4R, 5R) -5- (4-aminopyrrole [2,1-f ] [1,2,4] triazin-7-yl) -5-cyano-3, 4-dihydroxytetrahydrofuran-2-yl) methyl 12- (((S) -1- ((2S, 4R) -4-hydroxy-2- (((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) carbamoyl) pyrrolidin-1-yl) -3, 3-dimethyl-1-oxobutan-2-yl) amino) dodecanoate.

5. A pharmacologically or physiologically acceptable salt of a ritac compound of ridciclovir or GS-441524 as claimed in any one of claims 1 to 4.

6. Use of a procac compound of reidexilvir or GS-441524 as claimed in any one of claims 1 to 4 or a salt as claimed in claim 4 for the manufacture of an anti-EV 71 medicament or an RNA-dependent RNA polymerase inhibitor.

7. An anti-EV 71 pharmaceutical composition or an RNA-dependent RNA polymerase inhibitor characterized by: at least one ProTAC compound comprising Reidesciclovir or GS-441524 as claimed in any one of claims 1 to 4 or a pharmacologically or physiologically acceptable salt thereof.

8. The anti-EV 71 pharmaceutical composition or RNA-dependent RNA polymerase inhibitor according to claim 7, wherein: also comprises a pharmaceutically acceptable carrier or excipient.

9. A process for the preparation of the procac compound of reed-civir or GS-441524 as claimed in any one of claims 1 to 4, characterized in that:

the ProTAC compound of the Rideciclovir or the GS-441524 is divided into a PROTAC compound of series I Rideciclovir, a PROTAC compound of series II GS-441524, a PROTAC compound of series III GS-441524, a PROTAC compound of series IV GS-441524 and a PROTAC compound of series V GS-441524;

wherein, the PROTAC compound of the series I Ruideciclovir is a compound shown in a general formula (I), linker is an alkylene chain or an alkoxy chain with n =6-20, and E3 ligand is VHL with methyl;

the PROTAC compound of the series II GS-441524 is a compound shown as a general formula (II), wherein Linker is an alkylene chain with n =6-10, and E3 ligand is methyl-bearing VHL;

the PROTAC compound of the series III GS-441524 is a compound shown as a general formula (III), wherein Linker is an alkylene or alkoxy chain with n =6-20, and E3 ligand is methyl-carrying VHL or trans-methyl-carrying VHL;

the PROTAC compound of the series IV GS-441524 is a compound shown as a general formula (III), wherein Linker is an alkylene chain with n =6-20, and E3 ligand is lenalidomide, methyllenalidomide or benzodioxopiperidine;

the PROTAC compound of the series V GS-441524 is a compound shown as a general formula (III), linker is an alkylene chain with n =6-11, and E3 ligand is Boc lysine, 9-aminofluorene or RNF-114;

the preparation method of the PROTAC compound of series I Ruixiwei comprises the following steps: reacting a VHL derivative with an alkanoic acid substituent at the end and an alkyl chain or an alkoxy chain as a connecting chain with Reidesciclovir in a solvent under the conditions of HATU and DIPEA to obtain a PROTAC compound of the series I Reidesciclovir;

a preparation method of PROTAC compounds of series II GS-441524 comprises the following steps: reacting a VHL derivative with an alkanoic acid at the tail end and an alkyl chain as a connecting chain with GS-441524 in a solvent under the conditions of HATU and DIPEA to obtain a PROTAC compound of a series II GS-441524;

the method (1) comprises the following steps: GS-441524 reacts with 2, 2-dimethoxypropane and concentrated sulfuric acid in acetone to obtain a 2'-OH,3' -OH protected intermediate; reacting the intermediate with a VHL derivative with a single alkyl chain without tert-butyl ester in a solvent under the conditions of EDCI and DMAP to obtain a 2'-OH,3' -OH protected PROTAC compound of GS-441524, and removing a protecting group under an acidic condition to obtain a series III PROTAC compound of GS-441524;

the method (2) comprises the following steps: GS-441524 reacts with 2, 2-dimethoxypropane and concentrated sulfuric acid in acetone to obtain a 2'-OH,3' -OH protected intermediate; the intermediate and a VHL derivative with a single alkyl chain without tert-butyl ester react in a solvent under the conditions of EDCI and DMAP to obtain a 2'-OH,3' -OH protected PROTAC compound of GS-441524, and finally, the protecting group is removed under the acidic condition to obtain a PROTAC compound of series III GS-441524;

the method (3) comprises the following steps: a PROTAC compound of GS-441524 protected by 2'-OH,3' -OH is obtained by dissolving a methyl VHL derivative with a bromine atom at the end and a single alkyl chain as a connecting chain, a GS-441524 intermediate protected by 2'-OH,3' -OH, potassium carbonate and potassium iodide in a solvent for reflux reaction; finally, removing a protecting group under an acidic condition to obtain a PROTAC compound of the series III GS-441524;

the preparation method of the PROTAC compound of the series IV GS-441524 comprises the following 2 methods:

the method (1) comprises the following steps: reacting a CRBN derivative with an alkanoic acid at the end and an alkoxy connecting chain with a GS-441524 intermediate protected by 2'-OH,3' -OH in a solvent under EDCI and DMAP conditions to obtain a 2'-OH,3' -OH protected PROTAC compound of GS-441524, and removing a protecting group under an acidic condition to obtain a series of IV GS-441524 PROTAC compounds;

the method (2) comprises the following steps: the method comprises the following steps of reacting benzodiketopiperidine with an alkanoic acid at the end and an alkoxy connecting chain with a GS-441524 intermediate protected by 2'-OH,3' -OH in a solvent under EDCI and DMAP conditions to obtain a 2'-OH,3' -OH protected PROTAC compound of GS-441524, and removing a protecting group under an acidic condition to obtain a series III GS-441524 PROTAC compound;

a preparation method of PROTAC compounds of series V GS-441524 comprises the following 3 methods:

the method (1) comprises the following steps: reacting long-chain alkanoic acid with a terminal Boc protected amino group with a 2'-OH,3' -OH protected GS-441524 intermediate in a solvent under EDCI and DMAP conditions to obtain a 2'-OH,3' -OH protected GS-441524 Linker conjugate compound, then removing a protecting group under an acidic condition to obtain GS-441524 connected with a long alkyl chain and with a terminal naked amino group, and finally reacting with Boc protected lysine to obtain a series of V-441524 PROTAC compounds;

the method (2) comprises the following steps: reacting a 9-aminofluorene derivative with an alkanoic acid at the end and an alkoxy connecting chain with a GS-441524 intermediate protected by 2'-OH,3' -OH in a solvent under the conditions of EDCI and DMAP to obtain a ProTAC compound of GS-441524 protected by 2'-OH,3' -OH, and removing a protecting group under an acidic condition to obtain a PROTAC compound of a series of V GS-441524;

the method (3) comprises the following steps: an RNF-114 ligand with an alkanoic acid at the end and an alkoxy connecting chain and a GS-441524 intermediate protected by 2'-OH,3' -OH react in a solvent under EDCI and DMAP conditions to obtain a PROTAC compound of GS-441524 protected by 2'-OH,3' -OH, and finally, a protecting group is removed under acidic conditions to obtain a PROTAC compound of a series V GS-441524.

10. The process for preparing a procac compound of reidecivir or GS-441524 as claimed in claim 9, wherein:

the preparation method of the PROTAC compound of the series I comprises the following steps: the ratio of the amounts of the redexivir, the VHL derivative, the HATU and the DIPEA is 1.1;

the preparation method of the PROTAC compound of the series II comprises the following steps: the mass ratio of GS-441524, VHL derivative, HATU and DIPEA was 1.1;

in the preparation method of the PROTAC compound of the series III, the mass ratio of GS-441524, 2-dimethoxypropane and concentrated sulfuric acid is 1; the mass ratio of the 2'-OH,3' -OH protected GS-441524 intermediate, VHL derivative, EDCI, DMAP was 1;

the preparation method of the series IV PROTAC compound comprises the following steps: the mass ratio of the 2'-OH,3' -OH protected GS-441524 intermediate, the CRBN derivative, EDCI and DMAP in the method (1) is 1.1; the mass ratio of the 2'-OH,3' -OH protected GS-441524 intermediate, the benzodione piperidine derivative, EDCI, 2DMAP in method (2) is 1.1;

the preparation method of the PROTAC compound of the series V comprises the following steps: the mass ratio of the 2'-OH,3' -OH protected GS-441524 intermediate, the alkyl acid derivative of the terminal Boc amino group, EDCI and DMAP in process (1) is 1.1; the mass ratio of the 2'-OH,3' -OH protected GS-441524 intermediate, the 9-aminofluorene derivative, EDCI and DMAP in method (2) is 1.1; the mass ratio of the 2'-OH,3' -OH-protected GS-441524 intermediate, the RNF-114 ligand derivative, EDCI and DMAP in method (3) is 1.1.