CN114456211B - Peptoid compound and preparation method and application thereof - Google Patents

Peptoid compound and preparation method and application thereof Download PDF

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CN114456211B
CN114456211B CN202111030321.2A CN202111030321A CN114456211B CN 114456211 B CN114456211 B CN 114456211B CN 202111030321 A CN202111030321 A CN 202111030321A CN 114456211 B CN114456211 B CN 114456211B
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pharmaceutically acceptable
peptidomimetic
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alkyl
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CN114456211A (en
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蒋晟
肖易倍
谢幼华
倪勇
郝海平
张阔军
廖金标
武倩倩
王天雨
章翔宇
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China Pharmaceutical University
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    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
    • C07F9/02—Phosphorus compounds
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    • C07F9/6558—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom containing at least two different or differently substituted hetero rings neither condensed among themselves nor condensed with a common carbocyclic ring or ring system
    • C07F9/65583—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom containing at least two different or differently substituted hetero rings neither condensed among themselves nor condensed with a common carbocyclic ring or ring system each of the hetero rings containing nitrogen as ring hetero atom
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    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
    • C07F9/02—Phosphorus compounds
    • C07F9/547—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom
    • C07F9/6558—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom containing at least two different or differently substituted hetero rings neither condensed among themselves nor condensed with a common carbocyclic ring or ring system
    • C07F9/65586—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom containing at least two different or differently substituted hetero rings neither condensed among themselves nor condensed with a common carbocyclic ring or ring system at least one of the hetero rings does not contain nitrogen as ring hetero atom

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Abstract

The invention discloses a peptidomimetic compound shown in a general formula I, pharmaceutically acceptable salts thereof, tautomers thereof, stereoisomers thereof, metabolites thereof, metabolic precursors thereof or prodrugs thereof; the peptidomimetic compounds, pharmaceutically acceptable salts, tautomers, stereoisomers, metabolites, metabolic precursors or prodrugs thereof and the pharmaceutical compositions of the invention have wide application and can be prepared into medicaments for treating/preventing SARS-CoV, HBV, HCV, H1N1, ebola or SARS-CoV-2 virus infection diseases, and IC 50 Values optimally reach nanomolar concentration levels.

Description

Peptoid compound and preparation method and application thereof
Technical Field
The invention belongs to the field of medicines, and particularly relates to a peptidomimetic compound, and a preparation method and application thereof.
Background
SARS-CoV-2 is a highly pathogenic, pandemic zoonotic virus with symptoms ranging from asymptomatic disease to moderate and severe pneumonia, as well as life-threatening complications including hypoxic respiratory failure, acute respiratory distress syndrome, multiple system organ failure, and ultimately death.
Like other coronaviruses, SARS-Cov-2 is broken down to release the nucleocapsid and viral genome upon entry into the host cell. The host cell ribosomes translate the Open Reading Frames (ORFs) 1a and 1b of the viral genome into polyproteins (PP) PP1a and PP1b, respectively, encoding 16 nonstructural proteins (nsps), while the remaining ORFs encode structural and accessory proteins. 3C-like cysteine protease (3CLpro,nsp5,EC 3.4.22.69) and papain-like cysteine protease (PLpro, nsp3, EC3.4.22.46) are highly coordinated, catalyzing PP cleavage to form nsp2-16, thereby forming a replication-transcription complex (RTC); the loss of 3CLpro enzymatic activity results in a viral life cycle arrest, and thus 3CLpro is critical for viral replication and survival. In addition, studies have shown that 3CLpro can also cleave host immune related proteins, such as the human innate immune molecule STING.3CLpro is not only necessary to maintain the replication of the virus itself, but also can disrupt the immune system of the host cell, suppress the anti-infective immune response, and cause immune escape. Inhibition of 3CLpro is not only effective in killing coronaviruses but also in reducing immune imbalance in infected host cells. Given the importance of 3CLpro in the gene replication process of coronaviruses, the development of 3CLpro inhibitors would produce potent inhibitory or killing effects on coronaviruses, which makes 3CLpro an interesting target for antiviral chemotherapy.
Reported 3CLpro inhibitors include peptide inhibitors (see Kim et al (2012) biorg. Med. Chem. Lett.22:6952-6956; thibaut et al (2012) biochem. Pharmacol.83:185-192; van et al (2014) anti-viral res.103:17-24; international patent application publication Nos. WO 2005/113580, WO 2006/061714, WO 2017/114509, zhang et al (2020) Science 368:409-412, dai et al (2020) Science 368:1331-1335, qiao et al (2021) Science 371:1374-1378, jacobs et al (2012) J.Med. Chem.56:534-546, akaJi et al (2011) J.Med. Chem.54:7962-7973, zhang et al (2020) J.Med. Chem.63:4562-4578, hoffman et al (2020) J.Med. Chem.63:12725-12747) and nonpeptidic inhibitors, such as heterocyclic esters (see Lu et al (2006) J.Med. Chem. 49-5154-5161), pyrazoles (see Ramayam et al (2010) Bioorg. Chem.7854-7854), indigo derivatives (see also see FIGS. 40-3052, etc. J.30:35, see also WO 35-43).
Disclosure of Invention
The invention aims to: the invention aims to provide a peptidomimetic compound capable of inhibiting 3C-like cysteine protease; another object of the present invention is to provide a process for the preparation of a peptidomimetic compound; another object of the present invention is to provide the use of a peptidomimetic compound as a novel 3C-like cysteine protease inhibitor of RNA viruses; another object of the present invention is to provide a pharmaceutical composition of a peptidomimetic compound; it is another object of the present invention to provide the use of a peptidomimetic compound, a pharmaceutical composition for the manufacture of a medicament for the treatment of a disease associated with abnormal 3C-like cysteine proteases of RNA viruses.
The technical scheme is as follows: the invention relates to a peptoid compound shown in a general formula I, a pharmaceutically acceptable salt thereof, a tautomer thereof, a stereoisomer thereof, a metabolite thereof, a metabolic precursor thereof or a prodrug thereof, wherein the general formula I is as follows:
in formula I:
x is NR 1 R 2 、OR 1 Or (CH) 2 ) a (C＝O)NR 1 R 2 ；
a＝0-4；
Y is O, S or NH;
z is
n＝0-3；
R 1 、R 2 Independently selected from hydrogen, hydroxy, cyano, C 1-8 Alkyl, cyano (C) 1-8 Alkyl), amino (C) 1-8 Alkyl group, C 1-8 alkylamino-C 1-8 Alkyl, hydroxy (C) 1-8 Alkyl), carboxyl (C) 1-8 Alkyl group, C 2-8 Alkenyl, C 2-8 Alkynyl, C 1-8 alkoxy-C 1-8 Alkyl, unsubstituted or R 1-1 Substituted C 3-10 Cycloalkyl, unsubstituted or R 1-2 Substituted heteroaryl, unsubstituted or R 1-3 Substituted heterocycloalkyl, unsubstituted or R 1-4 Substituted C 3-10 Cycloalkyl- (C) 1-6 Alkyl) -, unsubstituted or R 1-5 Substituted heteroaryl- (C) 1-6 Alkyl) -, unsubstituted or R 1-6 Substituted heterocycloalkyl- (C) 1-6 Alkyl) -, unsubstituted or R 1-7 Substituted C 6-10 Aryl, unsubstituted or R 1-8 Substituted C 6-10 Aryl- (C) 1-6 Alkyl) -, or R 1 And R is 2 Together with the nitrogen atom to which they are attached form unsubstituted or R 1-9 Substituted heterocycloalkyl, or R 1 And R is 2 Together with the nitrogen atom to which they are attached form unsubstituted or R 1-10 Substituted heteroaryl;
R 1-1 ～R 1-6 respectively selected from hydroxy, cyano, amino, halogen, C 1-6 Alkyl, halo (C) 1-6 Alkyl), hydroxy (C) 1-6 Alkyl group, C 1-6 Alkoxy or C 1-6 One of the alkylamino groups;
R 1-7 and R is 1-8 Respectively selected from hydroxy, cyano, halogen, nitro, C 1-6 Alkyl, C of (2) 2-8 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, C 6-10 Aryloxy, heteroaryloxyRadical (C) 3-10 Cycloalkyl) -oxy, halo (C) 1-6 Alkyl), hydroxy (C) 1-6 Alkyl), amino (C) 1-6 Alkyl group, C 1-6 alkylamino-C 1-6 Alkoxy-, C 3-10 Cycloalkyl, C 3-10 Cycloalkyl- (C) 1-6 Alkyl) -, C 3-10 Cycloalkyl- (C) 1-6 Alkoxy), unsubstituted or R 1-1-1 Substituted C 6-10 Aryl, C 6-10 Aryl- (C) 1-6 Alkyl) -, unsubstituted or R 1-1-2 Substituted C 6-10 Aryl- (C) 1-6 Alkoxy) -, heterocycloalkyl- (C) 1-6 Alkyl) -, unsubstituted or R 1-1-3 Substituted heteroaryl, heteroaryl- (C) 1-6 Alkyl) -, heteroaryl- (C 1-6 Alkoxy) -, NR 1-1-4 R 1-1-5 、-(C＝O)R 1-1-6 、-(C＝O)NR 1 -1-7 R 1-1-8 、 -NR 1-1-9 (C＝O)R 1-1-10 、-(C＝O)OR 1-1-11 、-O(C＝O)R 1-1-12 、-(S＝O) 2 NR 1-1-13 R 1-1-14 、 -NR 1-1-15 (S＝O) 2 R 1-1-16 Or- (s=o) 2 R 1-1-17 One of the following;
R 1-1-1 、R 1-1-2 and R is 1-1-3 Respectively selected from C 1-4 Alkyl, hydroxy (C) 1-4 Alkyl), halogen, cyano, hydroxy, C 1-4 Alkylamino, C 1-4 Alkoxy or halo (C) 1-4 Alkyl);
R 1-1-4 ～R 1-1-17 respectively hydrogen or C 1-4 An alkyl group;
R 1-9 is hydroxy, amino, C 1-6 Alkyl, C 6-10 Aryl or C 3-10 Cycloalkyl;
R 1-10 is hydroxy, halogen, C 1-6 Alkyl, amino, halo (C) 1-6 Alkyl group, C 1-6 Alkoxy, C 1-6 Alkylamino, hydroxy (C) 1-6 Alkyl), amino (C) 1-6 Alkyl group, C 6-10 Aryl or C 3-10 Cycloalkyl;
R 3 selected from the group consisting of
R 4 Is hydrogen, unsubstituted or R 4-1 Substituted C 1-8 Alkyl, unsubstituted or R 4-1 Substituted C 1-8 Alkylamino, unsubstituted or R 4-1 Substituted C 1-8 Alkoxy, C 2-8 Alkenyl, C 2-8 Alkynyl, C 2-8 Carboxyl, C 2-8 Ester group, unsubstituted or R 4-2 Substituted C 3-10 Cycloalkyl, unsubstituted or R 4-5 Substituted heterocycloalkyl, unsubstituted or R 4-3 Substituted C 6-10 Aryl, unsubstituted or R 4-4 Substituted heteroaryl;
R 4-1 one of hydroxyl, sulfhydryl, cyano, amino, nitro, ester, carboxyl and halogen;
R 4-2 ～R 4-5 、R 5 respectively selected from hydroxy, mercapto, cyano, amino, nitro, ester, carboxyl, halogen and halo (C) 1-6 Alkyl), hydroxy (C) 1-8 Alkyl), mercapto (C) 1-8 Alkyl), amino (C) 1-8 Alkyl), cyano (C) 1-8 Alkyl), ester group (C) 1-8 Alkyl), carboxyl (C) 1-8 Alkyl group, C 1-8 Alkoxy, C 1-8 One of the alkylamino groups;
R 6 、R 7 independently selected from hydrogen, hydroxy, cyano, C 1-8 Alkyl, cyano (C) 1-8 Alkyl), amino (C) 1-8 Alkyl group, C 1-8 alkylamino-C 1-8 Alkyl, hydroxy (C) 1-8 Alkyl group, C 2-8 Alkenyl, C 2-8 Alkynyl, C 1-8 alkoxy-C 1-8 Alkyl, unsubstituted or R 4-1 Substituted C 3-10 Cycloalkyl, unsubstituted or R 4-2 Substituted heterocycloalkyl, unsubstituted or R 4-3 Substituted C 3-10 Cycloalkyl- (C) 1-6 Alkyl) -, unsubstituted or R 4-4 Substituted heterocycloalkyl- (C) 1-6 Alkyl) -, unsubstituted or R 4-5 Substituted C 6-10 Aryl- (C) 1-6 Alkyl group-, or R 6 And R is 7 Together with the nitrogen atom to which they are attached form unsubstituted or R 4-2 Substituted heterocycloalkyl;
further, in the above definition of the various moieties of formula I, the preferred groups for the various moieties are as follows:
x is NR 1 R 2 、OR 3 Or (CH) 2 ) a (C＝O)NR 1 R 2 Wherein NR is 1 R 2 Selected from any one of the following groups:
OR 3 selected from the following groups:
wherein U is 1 Selected from hydrogen, hydroxy, amino, halogen, C 1-8 Alkyl, trifluoromethyl, methoxy, trifluoromethoxy or hydroxymethyl;
U 2 -U 6 independently selected from hydrogen, C 1-8 Alkyl, hydroxy, amino, nitro, cyano, carboxyl, carboxylate, halogen, methoxy, trifluoromethyl, trifluoromethoxy;
U 7 selected from methyl, ethyl or isopropyl;
z isWherein->Selected from any one of the following groups:
selected from the following groups:
wherein U is 2 The same definition as above;
y is preferably O;
a is preferably 1;
R 3 selected from the following groups:
further, the compound shown in the formula I is any one of the following compounds:
a process for the preparation of the above-mentioned peptidomimetic compound, a pharmaceutically acceptable salt thereof, a tautomer thereof, a stereoisomer thereof, a metabolite thereof, a metabolic precursor thereof or a prodrug thereof, which process is any one of the following:
the method comprises the following steps: raw material S1 reacts with lithium bis (trimethylsilyl) amide and bromoacetonitrile to obtain a compound S2; adding cobalt chloride hexahydrate and sodium borohydride into the compound S2 to react to obtain a compound S3; the compound S3 is subjected to demethylation, then reacts with isobutyl chloroformate and triethylamine, reacts with diazomethane to obtain an intermediate, and reacts with dioxane solution of hydrogen chloride to obtain a compound S4; the raw material S5 reacts with triphosgene under alkaline condition to obtain an isocyanate intermediate, and reacts with another molecule of amine to obtain a compound S6; removing methyl ester from the compound S6 in the presence of lithium hydroxide to obtain a compound S7; condensing the compounds S4 and S7 under the condition of condensing agents N, N, N ', N' -tetramethyl-O- (7-azabenzotriazole-1-yl) hexafluoro urea phosphate and alkali N-methylmorpholine to obtain a compound S8; the compound S8 and the benzoyl formic acid generate corresponding ester in the presence of alkali cesium fluoride, and then alkaline hydrolysis is carried out to obtain a compound S9; the compound S9 reacts with N, N-diisopropyl phosphoramidite di-tert-butyl ester, and is oxidized by hydrogen peroxide to obtain a compound S10; acidifying the compound S10 by trifluoroacetic acid to obtain a compound (I-I);
the second method is as follows: reacting the compound S5, malonic acid with N, N, N ', N' -tetramethyl-O- (7-azabenzotriazole-1-yl) hexafluorophosphate urea and N-methylmorpholine to obtain a compound S11; reacting the compound S11 with another molecule of amine and N, N, N ', N' -tetramethyl-O- (7-azabenzotriazol-1-yl) hexafluorophosphate urea and N-methylmorpholine to obtain a compound S12; reacting the compound S12 with sodium hydroxide solution, and regulating the reaction product to be acidic by hydrochloric acid to obtain a compound S13; condensing the compounds S4 and S13 under the condition of condensing agents N, N, N ', N' -tetramethyl-O- (7-azabenzotriazole-1-yl) urea hexafluorophosphate and alkali N-methylmorpholine to obtain a compound S14; the compound S14 and the benzoyl formic acid generate corresponding ester in the presence of alkali cesium fluoride, and then alkaline hydrolysis is carried out to obtain a compound S15; the compound S15 reacts with N, N-diisopropyl phosphoramidite di-tert-butyl ester, and is oxidized by hydrogen peroxide to obtain a compound S16; acidifying the compound S16 by trifluoroacetic acid to obtain a compound (I-II);
the use of the above-mentioned peptidomimetic compounds as inhibitors of 3C-like cysteine proteases of RNA viruses.
A pharmaceutical composition comprising the above-described peptidomimetic compound, a pharmaceutically acceptable salt thereof, a tautomer thereof, a stereoisomer thereof, a metabolite thereof, a metabolic precursor thereof or a prodrug thereof, and a pharmaceutically acceptable carrier; in the pharmaceutical composition, the amount of the peptidomimetic compound, a pharmaceutically acceptable salt thereof, a tautomer thereof, a stereoisomer thereof, a metabolite thereof, a metabolic precursor thereof, or a prodrug thereof may be a therapeutically effective amount.
Use of a peptidomimetic compound as defined above, a pharmaceutically acceptable salt thereof, a tautomer thereof, a stereoisomer thereof, a metabolite thereof, a metabolic precursor thereof or a prodrug thereof, as defined above, in the manufacture of a medicament for the treatment of a disease associated with an abnormality of a 3C-like cysteine protease inhibitor of an RNA virus;
wherein the disease associated with abnormal 3C-like cysteine protease inhibitors of RNA viruses is a viral infection disease; the virus infection is one or more of SARS-CoV, HBV, HCV, H1N1, ebola and SARS-CoV-2. The application of the pharmaceutical composition in preparing medicines or vaccine adjuvants, wherein the medicines are used for treating viral infection; the vaccine adjuvant is used for treating virus infection;
the beneficial effects are that: compared with the prior art, the invention has the following remarkable advantages: (1) The small molecules and the derivatives thereof have high-efficiency inhibitory activity on the 3C-like cysteine protease of RNA viruses; (2) The analogue peptide compound, its derivative and medicine composition have wide application, and may be prepared into medicine for preventing and treating SARS-CoV, HBV, HCV, H1N1, ebola or SARS-CoV-2 virus infection diseases and IC 50 The value is optimal and can reach the nanomolar concentration level; (3) The preparation method of the compound is easy to operate and has wide applicability of reaction substrates.
Detailed Description
The technical scheme of the invention is further described below.
Example 1: synthesis of Compound 1
(S) -3- ((S) -4-methyl-2- (3-phenylurea) pentanamide) -2-oxo-4- ((S) -2-oxopyrrolidin-3-yl) butyl dihydrogen phosphate
Step one: synthesis of Compound S2
Raw material S1 (20 g,72 mmol) was weighed and dissolved in anhydrous tetrahydrofuran (250 mL), lithium bis (trimethylsilyl) amide (154 mL,154 mmol) was slowly added dropwise under nitrogen atmosphere at-78 ℃, after stirring for 1h, bromoacetonitrile (5.1 mL,76 mmol) solution of tetrahydrofuran (30 mL) was slowly added dropwise, after stirring for 3h, cold methanol (12 mL) solution and acetic acid (12 mL) solution of tetrahydrofuran (80 mL) were added to quench, the temperature was slowly raised to room temperature, the solvent was removed under reduced pressure, extracted with ethyl acetate (300 mL), washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, concentrated, made into sand, and purified by column chromatography (PE: ea=5:1) to give colorless liquid S2 (18 g, 77%). 1 H NMR(300MHz,Chloroform-d)δ5.12 (d,J＝8.8Hz,1H),4.44–4.28(m,1H),3.76(s,3H),3.75(s,3H),2.82(dq,J＝17.8, 4.8,3.3Hz,3H),2.25–2.06(m,2H),1.44(s,9H).
Step two: synthesis of Compound S3
The resulting compound S2 (10.6 g,33.7 mmol) was dissolved in methanol (250 mL), cobalt chloride hexahydrate (4.8 g,20.23 mmol) was added, and sodium borohydride (7.7 g,202.3 mmol) was added as a solid in portions at 0deg.C and reacted overnight at room temperature. The reaction was stopped, quenched with saturated ammonium chloride solution (100 mL), filtered through celite, the solvent was removed under reduced pressure, extracted with ethyl acetate (200 mL), washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, concentrated, sand-packed, and purified by column chromatography (PE: ea=1:2) to give compound S3 (4.8 g, 50%) as a white foamy solid. 1 H NMR(300MHz,Chloroform-d)δ6.12 (s,1H),5.49(s,1H),4.31(d,J＝10.7Hz,1H),3.74(s,3H),3.41–3.27(m,2H),2.56–2.32(m,2H),2.13(ddd,J＝14.4,10.7,3.7Hz,1H),1.98–1.75(m,2H),1.43(s, 9H).
Step three: synthesis of Compound S4
Compound S3 (4.5 g,15.7 mmol) was dissolved in tetrahydrofuran (40 mL), 4N aqueous sodium hydroxide solution (20 mL,78.5 mmol) was added, after stirring at room temperature for 6h, 4N hydrochloric acid was added to adjust pH to 3, extraction was performed three times with ethyl acetate (50 mL), washing was performed with saturated brine (40 mL), drying was performed with anhydrous sodium sulfate, and filtration was performed to concentrate a white solid (4.2 g, 98%) for the next reaction.
The white solid (3 g,11 mmol) obtained in the previous step was dissolved in tetrahydrofuran (20 mL) at 0deg.CTriethylamine (2.3 mL,16.5 mmol) and isobutyl chloroformate (1.7 mL,13.2 mmol) were slowly added dropwise, the solution became cloudy, and after completion of the reaction, an ether solution of diazomethane (54 mL,22 mmol) was added and reacted at room temperature for 16h. Extraction with ethyl acetate (50 mL), washing with saturated aqueous sodium bicarbonate (50 mL), saturated brine (50 mL), drying over anhydrous sodium sulfate, filtration, concentration, column chromatography gave a pale yellow solid (2.7 g, 84%). 1 H NMR(300MHz,DMSO-d 6 )δ7.64(s,1H),7.43(d,J＝8.2Hz,1H), 6.05(d,J＝9.9Hz,1H),4.03(d,J＝38.7Hz,1H),3.18–3.06(m,2H),2.30–2.06 (m,2H),2.05–1.79(m,1H),1.72–1.46(m,2H),1.39(s,9H).
The pale yellow solid (2.7 g,9.1 mmol) obtained in the above step was dissolved in an anhydrous dioxane solution (32 mL), and a dioxane solution (22 mL,91 mmol) of hydrogen chloride was slowly added dropwise at 0℃and stirred at room temperature for 3 hours. After completion of the reaction, white solid S4 (1.4 g, 62%) was obtained by filtration. 1 H NMR(400MHz,DMSO-d 6 )δ 8.69(s,3H),7.97(s,1H),4.91(dd,J＝17.2,10.1Hz,1H),4.77(dd,J＝17.2,7.0Hz,1H),4.31(d,J＝4.3Hz,1H),3.24–3.14(m,2H),2.61(p,J＝7.9Hz,1H),2.35– 2.26(m,1H),2.04–1.85(m,2H),1.78–1.61(m,1H).
Step four: synthesis of Compound S6
Triphosgene (2.1 g,6.9 mmol) was weighed and dissolved in anhydrous dichloromethane (20 mL), raw material S5 (1 g,6.9 mmol) was added under nitrogen protection at 0deg.C, triethylamine (1.9 mL,13.8 mmol) was slowly added dropwise after stirring for 5min, stirring at room temperature for 2h, after shifting to 0deg.C, aniline (0.8 mL,8.3 mmol) was slowly added dropwise in dichloromethane (20 mL), and the reaction was carried out at room temperature overnight. The reaction was stopped, quenched with saturated ammonium chloride solution (20 mL), filtered through celite, the solvent was removed under reduced pressure, extracted with dichloromethane (50 mL), washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, concentrated, sand-packed, and purified by column chromatography to give colorless liquid compound S6 (1.4 g, 79%). 1 H NMR(300MHz,Chloroform-d)δ7.30–7.22(m, 4H),7.09(dd,J＝12.6,7.2Hz,1H),4.49(t,J＝6.0Hz,1H),3.65(s,3H),1.82–1.78(m,2H),1.49–1.43(m,1H),0.93(s,3H),0.91(s,3H).
Step five: synthesis of Compound S7
The compound is preparedS6 (1.3 g,4.9 mmol) was dissolved in tetrahydrofuran (10 mL), 4N aqueous sodium hydroxide solution (6.1 mL,25 mmol) was added, after stirring at room temperature for 6h, 4N hydrochloric acid was added to adjust pH to 3, extraction was performed three times with ethyl acetate (20 mL), washing was performed with saturated brine (20 mL), drying was performed with anhydrous sodium sulfate, and filtration and concentration were performed to obtain S7 (1.1 g, 93%) as a white solid. 1 H NMR(300MHz,Chloroform-d)δ7.32 –7.23(m,4H),7.10(dd,J＝12.5,7.0Hz,1H),4.48(t,J＝5.8Hz,1H),1.85–1.79 (m,2H),1.48–1.43(m,1H),0.94(s,3H),0.92(s,3H).
Step six: synthesis of Compound S8
Compound S4 (0.96 g,4.0 mmol), compound S7 (1.0 g,4.0 mmol) were dissolved in N, N-dimethylformamide (20 mL), and urea N, N, N ', N' -tetramethyl-O- (7-azabenzotriazol-1-yl) hexafluorophosphate (1.7 g,4.4 mmol) and N-methylmorpholine (1.3 mL,12 mmol) were added at 0deg.C and reacted for 3h. Saturated ammonium chloride solution (10 mL) was added and quenched, extracted with ethyl acetate (20 mL. Times.3), washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and sand-packed, and purified by column chromatography to give compound S8 (1.1 g, 63%) as a white solid. 1 H NMR(300MHz,Chloroform-d)δ7.34 –7.26(m,4H),7.11(dd,J＝12.8,7.5Hz,1H),4.68–4.60(m,2H),4.39–4.34(m, 2H),3.25–3.14(m,2H),2.15–2.08(m,2H),2.00–1.92(m,3H),1.60(d,J＝12.9Hz,2H),1.39–1.34(m,1H),0.86(dd,J＝9.9,6.5Hz,6H).
Step seven: synthesis of Compound S9
Compound S8 (1.0 g,2.3 mmol) and benzoic acid (345 mg,2.3 mmol) were dissolved in DMF (10 mL), cesium fluoride (700 mg,4.6 mmol) was added and reacted at 60℃for 4h. Saturated ammonium chloride solution (10 mL) was added and quenched, extracted with ethyl acetate (20 mL. Times.3), washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to give a white solid (860 mg, 68%). 1 H NMR(300MHz,Chloroform-d)δ7.80(d,J＝8.2Hz,2H),7.51–7.45(m, 3H),7.38–7.30(m,4H),7.12(dd,J＝13.6,8.1Hz,1H),4.95–4.89(m,2H),4.38–4.33(m,2H),3.27–3.17(m,2H),2.14–2.08(m,2H),2.01–1.93(m,3H),1.62(d, J＝12.1Hz,2H),1.35–1.30(m,1H),0.85(dd,J＝9.5,5.5Hz,6H).
The white obtained in the previous step is mixedThe colored solid (850 mg,1.5 mmol) was dissolved in tetrahydrofuran (5 mL), 2N aqueous lithium hydroxide (1.5 mL,3.0 mmol) was added, and after stirring at room temperature for 2 hours, it was extracted with ethyl acetate (10 mL. Times.3), washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give S9 (530 mg, 85%) as a white solid. 1 H NMR(300MHz,Chloroform-d)δ7.31–7.22(m,4H),7.10(dd, J＝12.1,6.2Hz,1H),4.64–4.58(m,2H),4.35–4.30(m,2H),3.26–3.15(m,2H), 2.14–2.07(m,2H),1.99–1.91(m,3H),1.62(d,J＝12.5Hz,2H),1.38–1.33(m,1H),0.89(dd,J＝9.8,5.3Hz,6H).
Step eight: synthesis of Compound S10
Compound S9 (500 mg,1.2 mmol) and tetrazole (126 mg,1.8 mmol) were dissolved in tetrahydrofuran (5 mL), N-diisopropylphosphoramidite di-tert-butyl ester (0.6 mL,1.8 mmol) was added at 0deg.C, the mixture was reacted at room temperature for 6 hours, and then, the mixture was moved to 0deg.C, hydrogen peroxide (0.3 mL,2.4 mmol) was slowly added dropwise thereto, and the reaction was continued for 2 hours. Saturated sodium sulfite solution (10 mL) was added and quenched, extracted with ethyl acetate (10 mL. Times.3), washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, concentrated, sand-making, and purified by column chromatography to give S10 (640 mg, 88%) as a white solid. 1 H NMR(300MHz,Chloroform-d)δ7.35–7.28(m, 4H),7.14(dd,J＝12.0,6.9Hz,1H),4.86–4.79(m,2H),4.37–4.31(m,2H),3.29–3.19(m,2H),2.17–2.08(m,2H),2.00–1.92(m,3H),1.65(d,J＝10.6Hz,2H), 1.30–1.23(m,19H),0.88(dd,J＝10.2,6.4Hz,6H).
Step nine: synthesis of Compound 1
Compound S10 (610 mg,1.0 mmol) was dissolved in dichloromethane (5 mL), and trifluoroacetic acid (0.15 mL,2.0 mmol) was slowly added dropwise at 0deg.C and stirred at room temperature for 3h. Extraction with ethyl acetate (10 mL. Times.3), washing with saturated brine (10 mL), drying over anhydrous sodium sulfate, filtration, and concentration gave white solid 1 (375 mg, 76%). 1 H NMR(300MHz,Chloroform-d)δ7.32–7.24(m,4H),7.11(dd,J＝ 11.8,5.8Hz,1H),4.66–4.57(m,2H),4.35–4.29(m,2H),3.27–3.18(m,2H),2.12–2.05(m,2H),1.99–1.93(m,3H),1.60(d,J＝13.6Hz,2H),1.35–1.30(m,1H), 0.99(dd,J＝10.1,6.7Hz,6H).MS(EI,m/z):499(M + +1).
Compound (I-I) can be synthesized using the synthesis method of example 1:
the specific synthesized compounds are shown in table 1.
Table 1 compounds synthesized using the synthetic method of example 1
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Example 2: synthesis of Compound 61
(S) -3- ((S) -4-methyl-2- (3-oxo-3- (phenylamino) propanamido) pentanamido) -2-oxo-4- ((S) -2-oxopyrrolidin-3-yl) butyl dihydrophosphate
Step one: synthesis of Compound S11
Compound S5 (2.0 g,13.8 mmol), malonic acid (1.4 g,13.8 mmol) were dissolved in N, N-dimethylformamide (50 mL), and urea N, N, N ', N' -tetramethyl-O- (7-azabenzotriazol-1-yl) hexafluorophosphate (6.3 g,16.6 mmol) and N-methylmorpholine (4.5 mL,41.4 mmol) were added at 0deg.C and reacted for 3h. Saturated ammonium chloride solution (50 mL) was added and quenched, extracted with ethyl acetate (50 mL. Times.3), washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to give compound S11 (2.3 g, 71%) as a white solid. 1 H NMR(300MHz,Chloroform-d)δ4.56 (dd,J＝8.4,2.8Hz,1H),3.62(s,3H),3.11(s,2H),1.82–1.77(m,2H),1.45–1.39 (m,1H),0.92(dd,J＝10.2,6.4Hz,6H).
Step two: synthesis of Compound S12
Compound S11 (2.0 g,8.7 mmol), aniline (0.79 mL,8.7 mmol) were dissolved in N, N-dimethylformamide (30 mL), and urea N, N, N ', N' -tetramethyl-O- (7-azabenzotriazol-1-yl) hexafluorophosphate (4.0 g,10.4 mmol) and N-methylmorpholine (2.9 mL,26.1 mmol) were added at 0deg.C and reacted for 3h. Saturated ammonium chloride solution (20 mL) was added and quenched, extracted with ethyl acetate (40 mL. Times.3), washed with saturated brine (40 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to give compound S12 (2.2 g, 85%) as a white solid. 1 H NMR(300MHz,Chloroform-d)δ7.52 –7.46(m,2H),7.30–7.23(m,3H),4.51(dd,J＝8.9,2.4Hz,1H),3.63(s,3H),3.12 (s,2H),1.81–1.75(m,2H),1.45–1.38(m,1H),0.93(dd,J＝11.4,6.3Hz,6H).
Step three: synthesis of Compound S13
Compound S12 (2.0 g,6.5 mmol) was dissolved in tetrahydrofuran (15 mL), 1M sodium hydroxide solution (13.0 mL,13.0 mmol) was slowly added dropwise at 0deg.C, after 2h at room temperature, ethyl acetate (20 mL) was added, the aqueous phase was taken after delamination, 1M hydrochloric acid solution was added to adjust pH to acidity, and filtration and washing with water afforded compound 27 (1.7 g, 90%) as a white solid. 1 H NMR(300MHz,Chloroform-d)δ7.53 –7.43(m,2H),7.31–7.24(m,3H),4.52(dd,J＝10.3,4.7Hz,1H),3.09(s,2H), 1.82–1.72(m,2H),1.43–1.37(m,1H),0.99(dd,J＝11.0,6.0Hz,6H).
Step four: synthesis of Compound S14
Compound S13 (1.0 g,3.4 mmol), compound S4 (816 mg,3.4 mmol) were dissolved in N, N-dimethylformamide (20 mL), and urea N, N, N ', N' -tetramethyl-O- (7-azabenzotriazol-1-yl) hexafluorophosphate (1.5 g,4.1 mmol) and N-methylmorpholine (1.1 mL,10.2 mmol) were added at 0deg.C and reacted for 3h. Saturated ammonium chloride solution (20 mL) was added thereto, and the mixture was quenched, extracted with ethyl acetate (30 mL. Times.3), washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to give Compound S14 (910 mg, 56%) as a white solid. 1 H NMR(300MHz,Chloroform-d)δ 7.55–7.48(m,2H),7.33–7.27(m,3H),4.42–4.37(m,4H),3.41–3.35(m,2H), 3.10(s,2H),2.06–1.96(m,5H),1.79–1.72(m,2H),1.46–1.39(m,1H),0.97(dd,J＝11.3,6.3Hz,6H).
Step five: synthesis of Compound S15
Compound S14 (900 mg,1.9 mmol) and benzoic acid (282 mg,1.9 mmol) were dissolved in DMF (10 mL), cesium fluoride (577 mg,3.8 mmol) was added and reacted at 60℃for 4h. Saturated ammonium chloride solution (10 mL) was added to quench, extracted with ethyl acetate (20 ml×3), washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to give a white solid (698 mg, 62%). 1 H NMR(300MHz,Chloroform-d)δ7.81–7.77(m,2H),7.57–7.45(m,5H), 7.33–7.26(m,3H),5.24(dd,J＝8.3,3.4Hz,2H),4.41(dd,J＝6.4,2.3Hz,2H),3.44–3.38(m,2H),3.12(s,2H),2.07–1.99(m,5H),1.76–1.71(m,2H),1.45– 1.37(m,1H),0.95(dd,J＝10.1,5.3Hz,6H).
The white solid (690 mg,1.2 mmol) obtained in the above step was dissolved in tetrahydrofuran (5 mL), 2N aqueous lithium hydroxide solution (1.2 mL,2.4 mmol) was added, and after stirring at room temperature for 2 hours, it was extracted with ethyl acetate (10 mL. Times.3), washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give white solid S9 (491 mg, 89%). 1 H NMR(300MHz,Chloroform-d)δ7.54–7.47(m,2H),7.34– 7.26(m,3H),4.96(dd,J＝8.9,2.6Hz,2H),4.39–4.31(m,2H),3.43–3.35(m,2H),3.08(s,2H),2.07–1.98(m,5H),1.76–1.70(m,2H),1.47–1.41(m,1H),0.98(dd, J＝11.7,5.4Hz,6H).
Step eight: synthesis of Compound S16
Compound S15 (490 mg,1.1 mmol) and tetrazole (91 mg,1.3 mmol) were dissolved in tetrahydrofuran (5 mL), N-diisopropylphosphoramidite di-tert-butyl ester (0.5 mL,1.6 mmol) was added at 0deg.C, the mixture was reacted at room temperature for 6 hours, and then, the mixture was moved to 0deg.C, and hydrogen peroxide (0.3 mL,2.2 mmol) was slowly added dropwise thereto, followed by further reaction for 2 hours. Saturated sodium sulfite solution (10 mL) was added and quenched, extracted with ethyl acetate (10 mL. Times.3), washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, concentrated, sand-making, and purified by column chromatography to give S16 (602 mg, 84%) as a white solid. 1 H NMR(300MHz,Chloroform-d)δ7.56–7.48(m, 2H),7.31–7.23(m,3H),5.12(dd,J＝8.6,2.7Hz,2H),4.40–4.33(m,2H),3.41–3.36(m,2H),3.02(s,2H),2.05–1.97(m,5H),1.75–1.69(m,2H),1.42–1.37(m, 1H),1.21(s,18H),0.95(dd,J＝11.0,5.5Hz,6H).
Step nine: synthesis of Compound 61
Compound S16 (600 mg,0.9 mmol) was dissolved in dichloromethane (5 mL), and trifluoroacetic acid (0.1 mL,1.3 mmol) was slowly added dropwise at 0deg.C, followed by stirring at room temperature for 3h. Extraction with ethyl acetate (10 mL. Times.3), washing with saturated brine (10 mL), drying over anhydrous sodium sulfate, filtration, and concentration gave white solid 1 (301 mg, 62%). 1 H NMR(300MHz,Chloroform-d)δ7.59–7.50(m,2H),7.32–7.26(m, 3H),5.11(dd,J＝8.9,2.5Hz,2H),4.42–4.36(m,2H),3.42–3.39(m,2H),3.04(s,2H),2.06–1.97(m,5H),1.74–1.68(m,2H),1.46–1.39(m,1H),0.96(dd,J＝11.7, 5.7Hz,6H).MS(EI,m/z):541(M + +1).
Compound (I-II) can be synthesized using the synthesis method of example 2:
the specific synthesized compounds are shown in table 2.
TABLE 2 Compounds synthesized using the synthetic method of example 2
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Example 3: tablet preparation
Compound 1 (50 g), hydroxypropylmethyl cellulose E (150 g), starch (200 g), povidone (appropriate amount) and magnesium stearate (1 g) prepared in example 1 were mixed, granulated, and tableted.
In addition, the compounds prepared in examples 1-2 can be prepared into capsules, powders, granules, pills, injections, syrups, oral liquids, inhalants, ointments, suppositories or patches and the like by endowing different pharmaceutical excipients according to the conventional preparation method of pharmacopoeia 2015.
Example 4: drug treatment CPE observations
Detection reagent: SARS-CoV-2 virus (generation: P6; titer: 2X 10) 5 TCID 50 Per mL), madin-Darby canine kidney (MDCK) cells were obtained from ATCC, influenza a/Hawaii/70/2019 (H1N 1), influenza a/Hong Kong/45/2019 (H3N 2) from national influenzaThe center provides virus, DMEM basal medium (Gibco), fetal bovine serum (Gibco), penicillin-streptomycin diabody (bioend), 0.25% pancreatin-EDTA.
Test procedure:
1) Inoculating cells: vero-E6 cells in the logarithmic growth phase were taken and digested with 0.25% pancreatin-EDTA and counted to obtain cell densities of: 1X 10 6 Cell suspension of individual/mL; taking 4mL of the above cells, adding 6mL of complete medium (DMEM with 10% FBS), and preparing into 4×10 cell density 5 Cell suspensions at a volume of each mL were seeded into 96-well plates at 100. Mu.L per well, 4X 10 cells per well 4 And each. Placing into a carbon dioxide incubator at 37 ℃ for continuous culture overnight.
2) Pretreatment of cell medicine: before virus infection, the drug was diluted to the corresponding concentration using a maintenance medium (DMEM with 2% fbs), 100 μl of the medium containing the drug of the corresponding concentration was added to each well, and the culture was continued in a carbon dioxide incubator at 37 ℃ for 1 hour.
3) Test drug dilution: adding 60 mu L of diluted medicine with 2 times of final concentration into each hole, setting cell control, and adding 120 mu L of maintenance medium; virus control, 60 μl of maintenance medium was added.
4) Virus dilution: virus stock droplets were 2.5X10 size 5 TCID 50 Per mL, collecting 200 μl of virus stock solution, adding 25mL of maintenance medium, mixing thoroughly, diluting the virus to 100TCID 50 /50μL。
5) And (3) dropwise adding viruses: hanging drop virus (except for cell control) was added to a 96-well plate at a volume of 60 μl/well, and the final virus-drug mix was 120 μl.
6) After the added virus-antibody was mixed on a shaker, the supernatant (100. Mu.L) of the cell-inoculated culture plate was aspirated, and then 100. Mu.L/well of the virus-drug mixture was aspirated and added thereto.
7) Drug antiviral ability was observed according to cytopathic effect: the cells were placed in a CO2 incubator at 37℃for 48 hours, and cytopathic effect was observed using an inverted microscope, and the results were recorded and statistically analyzed.
8) Inoculating cells: huh 7 cells in the logarithmic growth phase were taken, digested with 0.25% pancreatin-EDTA, and inoculated into well plates.
9) And (3) drug treatment: huh 7 cells were treated with the compound of formula I at a drug treatment concentration of 20. Mu.M, respectively.
10 Collecting cells: cells were collected 48 hours later and protein expression was detected by Western blot.
The results show that the S1-S393 compounds inhibited cytopathic effects of SARS-CoV-2 infected Vero E6 cells better than the control. The S1-S393 compound has a certain inhibiting effect on SARS-CoV-2.
Example 5: anti-influenza virus experiments
1) Cell and virus strain
Madin-Darby canine kidney (MDCK) cells were obtained from ATCC, influenza A/Hawaii/70/2019 (H1N 1), influenza A/Hong Kong/45/2019 (H3N 2) were supplied by the national influenza center
2) Experimental procedure
MDCK cells were seeded in 96-well plates with a cell count of 2X 10 per well 5 After cells grow into monolayers, they are infected with a certain amount of virus (100 TCID 50 ) After adsorption for 2h at 37 ℃, washing with MEM, changing with a maintenance solution of small molecular compounds (0-500 mug/mL) with different concentration gradients, and incubating at 37 ℃ in the environment of 5% carbon dioxide, and simultaneously setting a virus control group and a normal cell control group without the tested small molecular compounds. When cytopathic effect (CPE) of the virus control group reached 4+, CPE results for each group were observed and recorded. Antiviral Activity Reed is used&Muench method calculation, EC 50 ＝ Antilog[A+(50-B)/(C-B)×D]Wherein A: log < 50% cumulative inhibitor drug concentration; b: an accumulated inhibition of < 50%; c: an accumulated inhibition of > 50%; d: log dilution factor.
Example 6: anti-SARS-CoV-2 Virus experiment
1) Cell and virus strain
African green monkey kidney cells (Vero E6) were obtained from ATCC,2019BetaCoV/Wuhan/WIV04/2019 isolated from the institute of Marhan virus, national academy of sciences
2) Experimental procedure
Vero E6 cells were seeded in 96-well plates, 3X 10 per well 5 The individual cells were cultured overnight at 37℃in an incubator with 5% carbon dioxide. After cells grown to a monolayer, the cells were washed once with PBS, SARS-CoV-2 virus (moi=0.03) was added, the virus solution was adsorbed for 2 hours, and after washing 3 times with PBS, 2% low melting point agarose-DMEM (4% fbs) medium containing gradient diluted small molecule compounds was added. Culturing in a culture box with 5% carbon dioxide at 37deg.C for 4 days, fixing with 4% paraformaldehyde for 15min, cleaning for 3 times, adding 0.8% crystal violet for dyeing for 10min, cleaning for three times, and oven drying. Picture collection was performed using an enzyme-linked fluorescence spot analyzer (CTL, immunospot S6 Universal) and plaques were counted. Dose response curves were plotted from the number of plaques, and half-effective concentrations EC were calculated 50 。
Cytotoxicity test
Taking MDCK cells and Vero cells in exponential growth phase, and seeding in 96-well plates with 2×10 cells per well 4 The cells were then given the small molecule compound to be tested at a mass concentration ranging from 0-2000 μg/mL and incubated in an incubator at 37℃for 2 days with 5% carbon dioxide. Toxicity test of MDCK cells and Vero cells adopts CPE method, and half cytotoxicity concentration CC of tested small molecule compound on cells 50 Using Reed&Muench method calculation, CC 50 ＝Antilog[A+(50-B)/(C-B)×D]Wherein A: log < 50% cumulative inhibitor drug concentration; b: an accumulated inhibition of < 50%; c: an accumulated inhibition of > 50%; d: log dilution factor.
Table 3 shows the activity and cytotoxicity results of the compounds of the invention against influenza virus, SARS-CoV-2 virus:
wherein si=cc 50 /EC 50 ；
A:EC 50 <1μM,B:EC 50 ＝10μM-1μM,C:EC 50 >10μM；
α:SI>50,β:SI＝1-50,γ:SI<1；
TABLE 3 Activity and cytotoxicity of Compounds 1-80 against H1N1, H3N2 and SARS-CoV-2
The above-mentioned cell experiment proves that the compound of the present invention can effectively inhibit influenza virus and SARS-CoV-2 virus, and can also have the same result for other RNA viruses, and the series of compounds can be used in the preparation of anti-RNA virus medicines.

Claims (8)

1. A peptidomimetic compound of the general formula I:
in formula I:
x is NR 1 R 2 、OR 3 ' or (CH) 2 ) a (C＝O)NR 1 R 2 ；
Wherein NR is 1 R 2 Selected from any one of the following groups:
OR 3 ' is selected from the following groups:
wherein U is 1 Selected from hydrogen, hydroxy, amino, halogen, C 1-8 Alkyl, trifluoromethyl, methoxy, trifluoromethoxy or hydroxymethyl;
U 2 -U 6 independently selected from hydrogen, C 1-8 Alkyl, hydroxy, amino, nitro, cyano, carboxyl, carboxylate, halogen, methoxy, trifluoromethyl, trifluoromethoxy;
U 7 selected from methyl, ethyl or isopropyl;
z isWherein->Selected from any one of the following groups:
a＝0-4；
y is O, S or NH;
R 3 is that
2. The peptidomimetic compound of claim 1, wherein Y is O.
3. The peptidomimetic compound, pharmaceutically acceptable salt thereof, of claim 1, wherein a is 1.
4. The peptoid compound of claim 1, pharmaceutically acceptable salts thereof, having one of the following structural formulas:
5. a method for preparing a peptidomimetic compound, a pharmaceutically acceptable salt thereof, characterized in that the method comprises the steps of:
raw material S1 reacts with lithium bis (trimethylsilyl) amide and bromoacetonitrile to obtain a compound S2;
adding cobalt chloride hexahydrate and sodium borohydride into the compound S2 to react to obtain a compound S3;
the compound S3 is reacted with isobutyl chloroformate and triethylamine to obtain an intermediate, and the intermediate is reacted with dioxane solution of hydrogen chloride to obtain a compound S4;
the raw material S5 reacts with triphosgene under alkaline condition to obtain an isocyanate intermediate, and reacts with another molecule of amine to obtain a compound S6;
removing methyl ester from the compound S6 in the presence of lithium hydroxide to obtain a compound S7;
condensing the compounds S4 and S7 under the condition of condensing agents N, N, N ', N' -tetramethyl-O- (7-azabenzotriazole-1-yl) hexafluoro urea phosphate and alkali N-methylmorpholine to obtain a compound S8;
the compound S8 and the benzoyl formic acid generate corresponding ester in the presence of alkali cesium fluoride, and then alkaline hydrolysis is carried out to obtain a compound S9; the compound S9 reacts with N, N-diisopropyl phosphoramidite di-tert-butyl ester, and is oxidized by hydrogen peroxide to obtain a compound S10;
acidifying the compound S10 by trifluoroacetic acid to obtain a compound (I-I);
wherein NR is 1 R 2 And R is 4 Is defined as in claim 1.
6. A method for preparing a peptidomimetic compound, a pharmaceutically acceptable salt thereof, characterized in that the method comprises the steps of:
reacting the compound S5, malonic acid with N, N, N ', N' -tetramethyl-O- (7-azabenzotriazole-1-yl) hexafluorophosphate urea and N-methylmorpholine to obtain a compound S11;
reacting the compound S11 with another molecule of amine and N, N, N ', N' -tetramethyl-O- (7-azabenzotriazol-1-yl) hexafluorophosphate urea and N-methylmorpholine to obtain a compound S12;
reacting the compound S12 with sodium hydroxide solution, and regulating the reaction product to be acidic by hydrochloric acid to obtain a compound S13;
condensing the compounds S4 and S13 under the condition of condensing agents N, N, N ', N' -tetramethyl-O- (7-azabenzotriazole-1-yl) urea hexafluorophosphate and alkali N-methylmorpholine to obtain a compound S14;
the compound S14 and the benzoyl formic acid generate corresponding ester in the presence of alkali cesium fluoride, and then alkaline hydrolysis is carried out to obtain a compound S15;
the compound S15 reacts with N, N-diisopropyl phosphoramidite di-tert-butyl ester, and is oxidized by hydrogen peroxide to obtain a compound S16;
acidifying the compound S16 by trifluoroacetic acid to obtain a compound (I-II);
wherein,may be selected from any one of the following groups:
R 4 、U 1 、U 2 、U 3 、U 4 、U 5 and U 6 Is as defined in claim 1.
7. A pharmaceutical composition comprising a peptidomimetic or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 4 and a pharmaceutically acceptable carrier.
8. Use of a peptidomimetic compound according to any one of claims 1 to 4, or a pharmaceutically acceptable salt thereof, for the manufacture of an antiviral medicament, said virus being one or more of H1N1 or SARS-CoV-2.
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