CN112724156A

CN112724156A - Polycyclic pyridone derivative, pharmaceutical composition and application thereof

Info

Publication number: CN112724156A
Application number: CN202011622929.XA
Authority: CN
Inventors: 蒋宇扬; 唐林; 张存龙; 吴伟彬; 李安琪; 陈大伟
Original assignee: SHENZHEN KUNJIAN INNOVATIVE DRUG RESEARCH INSTITUTE
Current assignee: Shenzhen Berry Biomedical Technology Co ltd
Priority date: 2020-12-30
Filing date: 2020-12-30
Publication date: 2021-04-30
Anticipated expiration: 2040-12-30
Also published as: WO2022143424A1; CN112724156B

Abstract

The invention belongs to the field of medicines, and relates to a polycyclic pyridone derivative, a pharmaceutical composition and application thereof. The polycyclic pyridone derivative is a compound shown in a formula (I) or a stereoisomer, a tautomer, a nitrogen oxide, a solvate, a metabolite, a pharmaceutically acceptable salt or a prodrug of the compound shown in the formula (I). Compared with the existing similar compounds, the compound of the invention not only can well inhibit the replication of influenza virus, but also has lower cytotoxicity.

Description

Polycyclic pyridone derivative, pharmaceutical composition and application thereof

Technical Field

The invention belongs to the field of medicines, and particularly relates to a polycyclic pyridone derivative, a pharmaceutical composition containing the polycyclic pyridone derivative, and applications of the polycyclic pyridone derivative and the pharmaceutical composition.

Background

Influenza is mainly caused by influenza virus infection, which can result in three to five million hospitalized cases and 29 to 65 million death cases per year on average. Influenza a viruses are often responsible for major outbreaks of influenza. However, in recent years, with the emergence of highly invasive virus strains such as H1N1, H5N1 and H7N9, and the emergence of more and more resistant virus strains, the existing anti-influenza drugs have been unable to meet the needs of people. Therefore, it is necessary to develop anti-influenza drugs having a novel mechanism of action.

The life cycle of influenza virus is a complex biological process that can be divided into the following steps: (I) binding of the virus to host cell surface receptors; (II) the virus is endocytosed into the host cell, and the viral membrane and endosomal membrane are fused; (III) viral ribonucleoproteins (vRNPs) are released into the cytoplasm and then transported into the nucleus; (IV) transcription and replication of viral RNA; (V) export of mRNA and vRNP from the nucleus, synthesis of associated proteins; (VI) the assembly of progeny virus, released from the cell by budding. Viral proteins and host cell proteins involved in the influenza virus infection cycle are important targets for the development of anti-influenza virus drugs. A number of anti-influenza drugs have been developed against these targets.

Small molecule anti-influenza virus preparations are mainly divided into five categories: (I) m2 ion channel protein inhibitors (amantadine, rimantadine, etc.); (II) Neuraminidase (NA) inhibitors (zanamivir, oseltamivir, peramivir, ranimivir, etc.); (III) Hemagglutinin (HA) inhibitors (Arbidol, nitazoxanide, etc.); (IV) inhibitors of RNA-dependent RNA polymerase (RdRp) (Favipiravir, Barosavir, etc.); (V) inhibitors of Nucleoprotein (NP) (naproxen et al). To date, only a few anti-influenza virus drugs have been approved for marketing.

Currently, the major anti-influenza drug used clinically is neuraminidase inhibitor (NAI). However, in recent years, drug-resistant influenza virus mutant strains are continuously appeared, which not only reduce the clinical curative effect of anti-influenza drugs, but also further threaten the public health safety. Therefore, the development of anti-influenza drugs with a new mechanism has important significance and wide application prospect.

Barosavir (Baloxavir marboxil), which is sold under the name Xofluza, is approved by the FDA to be marketed in 2018 and 10 months, and is the first anti-influenza drug with a new action mechanism in the last twenty years. It was developed by japan salt bisense and was first approved for marketing in japan in 2018 and 2 months. Unlike neuraminidase inhibitors, baroxavir, a novel cap-dependent endonuclease (CEN) inhibitor, which is present in the polymerase acid Protein (PA) subunit of influenza a and b viruses, prevents progeny virions from being released from infected host cells. Viral mRNA transcription depends on a unique "cap-snatching" mechanism: PB2 bound to the 5'-cap transcript in the host, and the cap-dependent endonuclease in PA endonucleases were used at a position approximately 12 nucleotides from 5' -cap, and the cleaved cap structure was used as a primer for viral mRNA transcription. Thus, inhibition of this endonuclease can inhibit mRNA transcription, and thus viral replication. In addition, compared with oseltamivir, the baloxavir has the advantages that treatment is not needed (oseltamivir is continuously treated for 5 days), only once-a-day administration is needed, the disease symptom relieving time is not inferior to that of oseltamivir, the virus titer and safety are superior to those of oseltamivir, and the like. This will break the pattern that neuraminidase inhibitor dominates the influenza market for nearly 20 years, provide a brand new therapeutic mechanism for influenza, and get the industry's extensive attention.

It is a further object in the art to optimize based on balosavir in order to obtain a more active anti-influenza drug.

Disclosure of Invention

The invention aims to provide a novel compound serving as an influenza virus RNA polymerase inhibitor, and more particularly, the invention provides a novel compound serving as a cap-dependent endonuclease inhibitor of influenza virus, and the compound and a composition thereof can be used for preventing, treating or relieving symptoms of patients infected by the influenza virus. Compared with the existing similar compounds, the compound of the invention not only can well inhibit the replication of influenza virus, but also has lower cytotoxicity, and the anti-influenza virus activity of one compound is better than that of the similar compound on the market. Therefore, compared with the existing similar compounds, the compound provided by the invention has better influenza virus activity.

In order to achieve the above object, a first aspect of the present invention provides a polycyclic pyridone derivative which is a compound represented by formula (I) or a stereoisomer, a tautomer, a nitrogen oxide, a solvate, a metabolite, a pharmaceutically acceptable salt of the compound represented by formula (I), or a prodrug thereof;

wherein,

p is hydrogen, benzyl, n-hexyl or P forming a prodrug^RA group;

A₁is CR_1AR_1BS or O;

A₂is CR_2AR_2BS or O;

A₃is CR_3AR_3BS or O;

A₄each independently is CR_4AR_4BS or O;

and, from A₁、A₂、A₃N number of A₄And A is₁Adjacent nitrogen atom and to A₄The number of hetero atoms in ring-forming atoms of rings formed by adjacent carbon atoms is 1 or 2;

R_1Aand R_1BEach independently hydrogen, halogen, alkyl, haloalkyl, alkoxy, or phenyl;

R_2Aand R_2BEach independently hydrogen, halogen, alkyl, haloalkyl, alkoxy, or phenyl;

R_3Aand R_3BEach independently hydrogen, halogen, alkyl, haloalkyl, alkoxy, or phenyl;

R_4Aand R_4BEach independently hydrogen, halogen, alkyl, haloalkyl, alkoxy, or phenyl;

R_3Aand R_3BOptionally taken together with adjacent carbon atoms to form a carbocyclic or heterocyclic ring;

n is 1 or 2;

R₁and R₂Each independently is hydrogen, halogen, hydroxy, alkyl, alkoxy, or haloalkyl;

P^Ris at least one of the groups of formulae a) to ac) selected from:

a)-C(＝O)-P^R0；

b)-C(＝O)-P^R1；

c)-C(＝O)-L-P^R1；

d)-C(＝O)-L-O-P^R1；

e)-C(＝O)-L-O-L-O-P^R1；

f)-C(＝O)-L-O-C(＝O)-O-P^R1；

g)-C(＝O)-O-P^R2；

h)-C(＝O)-N(-K)(P^R2)；

i)-C(＝O)-O-L-O-P^R2；

j)-C(P^R3)₂-O-P^R4；

k)-C(P^R3)₂-O-L-O-P^R4；

l)-C(P^R3)₂-O-C(＝O)-P^R4；

m)-C(P^R3)₂-O-C(＝O)-O-P^R4；

n)-C(P^R3)₂-O-C(＝O)-N(-K)-P^R4；

o)-C(P^R3)₂-O-C(＝O)-O-L-O-P^R4；

p)-C(P^R3)₂-O-C(＝O)-O-L-N(P^R4)₂；

q)-C(P^R3)₂-O-C(＝O)-N(-K)-L-O-P^R4；

r)-C(P^R3)₂-O-C(＝O)-N(-K)-L-N(P^R4)₂；

s)-C(P^R3)₂-O-C(＝O)-O-L-O-L-O-P^R4；

t)-C(P^R3)₂-O-C(＝O)-O-L-N(-K)-C(＝O)-P^R4；

u)-C(P^R3)₂-O-P(＝O)(-P^R5)₂；

v)-C(P^R3)₂-P^R6)₂；

w)-C(＝N⁺(P^R7)₂)(-N(P^R7)₂)；

x)-C(P^R3)₂-C(P^R3)₂-C(C＝O)-O-P^R2；

y)-C(P^R3)₂-N(-K)-C(C＝O)-O-P^R2；

z)-P(＝O)(-P^R8)(-P^R9)；

aa)-S(＝O)₂-P^R10；

ab)-P^R11；

ac)-C(P^R3)₂-C(P^R3)₂-O-P^R2；

wherein L is a linear or branched alkylene group or a linear or branched alkenylene group;

k is hydrogen or alkyl optionally substituted by substituent group a;

P^R0is alkyl optionally substituted with substituent A or alkenyl optionally substituted with substituent A;

P^R1is carbocyclyl optionally substituted with substituent A, heterocyclyl optionally substituted with substituent A, alkylamino optionally substituted with substituent A, or alkylthio optionally substituted with substituent A;

P^R2is alkyl optionally substituted with substituent A, carbocyclyl optionally substituted with substituent A, heterocyclyl optionally substituted with substituent A, carbocycloalkyl optionally substituted with substituent A, heterocycloalkyl optionally substituted with substituent A, or trialkylsilyl;

P^R3each independently is hydrogen, alkyl;

P^R4each independently is alkyl optionally substituted with substituent A, carbocyclyl optionally substituted with substituent A, heterocyclyl optionally substituted with substituent A, alkylamino optionally substituted with substituent A, carbocycloalkyl optionally substituted with substituent A, heterocycloalkyl optionally substituted with substituent A, or trialkylsilyl;

P^R5each independently is OBn;

P^R6is carbocyclyl optionally substituted with substituent A, or heterocyclyl optionally substituted with substituent A;

P^R7each independently is alkyl optionally substituted with substituent A;

P^R8is alkoxy optionally substituted with substituent a;

P^R9is alkoxy optionally substituted with substituent A, alkylamino optionally substituted with substituent A, carbocycloxy optionally substituted with substituent A, heterocyclooxy optionally substituted with substituent A, carbocycloamino optionally substituted with substituent A, or heterocycloamino optionally substituted with substituent A;

and, P^R8And P^R9Optionally taken together with the adjacent phosphorus atom to form a heterocyclic ring optionally substituted with substituent A;

P^R10is alkyl optionally substituted with substituent A, carbocyclyl optionally substituted with substituent A, heterocyclyl optionally substituted with substituent A, carbocycloalkyl optionally substituted with substituent A, or heterocycloalkyl optionally substituted with substituent A;

P^R11is alkyl optionally substituted with substituent A, alkenyl optionally substituted with substituent A, carbocyclyl optionally substituted with substituent A, or heterocyclyl optionally substituted with substituent A;

the substituent A is selected from at least one of the group consisting of: oxo, alkyl, hydroxyalkyl, amino, alkylamino, carbocyclyl, heterocyclyl, carbocycloalkyl, alkylcarbonyl, halogen, hydroxy, carboxy, alkylcarbonylamino, alkylcarbonylaminoalkyl, alkylcarbonyloxy, alkoxycarbonyl, alkoxycarbonylalkyl, alkoxycarbonyloxy, alkylaminocarbonyloxy, alkylaminoalkyl, alkoxy, cyano, nitro, azido, alkylsulfonyl, trialkylsilyl, and phosphoryl.

Specifically, the polycyclic pyridone derivative is a compound shown as a formula (II) or a stereoisomer, a tautomer, a nitrogen oxide, a solvate, a metabolite, a pharmaceutically acceptable salt or a prodrug thereof,

wherein,

p is hydrogen, benzyl, n-hexyl or P forming a prodrug^RA group;

P^Ris at least one selected from the following groups:

a)-C(＝O)-P^R0；

b)-C(＝O)-P^R1；

g)-C(＝O)-O-P^R2；

h)-C(＝O)-N(-K)(P^R2)；

i)-C(＝O)-O-L-O-P^R2；

l)-C(P^R3)₂-O-C(＝O)-P^R4；

m)-C(P^R3)₂-O-C(＝O)-O-P^R4；

o)-C(P^R3)₂-O-C(＝O)-O-L-O-P^R4；

v)-C(P^R3)₂-P^R6)₂；

x)-C(P^R3)₂-C(P^R3)₂-C(C＝O)-O-P^R2；

y)-C(P^R3)₂-N(-K)-C(C＝O)-O-P^R2；

z)-P(＝O)(-P^R8)(-P^R9)；

wherein L is a linear or branched alkylene group;

k is hydrogen or alkyl optionally substituted with substituent A;

P^R0is alkyl optionally substituted with substituent A;

P^R1is carbocyclyl optionally substituted with substituent A, heterocyclyl optionally substituted with substituent A;

P^R2is alkyl optionally substituted with substituent A, carbocyclyl optionally substituted with substituent A, heterocyclyl optionally substituted with substituent A, carbocycloalkyl optionally substituted with substituent A, heterocycloalkyl optionally substituted with substituent A;

P^R3each independently is hydrogen, alkyl;

P^R4is alkyl optionally substituted with substituent A, carbocyclyl optionally substituted with substituent A, heterocyclyl optionally substituted with substituent A;

P^R6is carbocyclyl optionally substituted with substituent A, heterocyclyl optionally substituted with substituent A;

P^R8is alkoxy optionally substituted with substituent a;

P^R9is alkoxy optionally substituted by substituent A, alkylamino optionally substituted by substituent A, carbocyloxy optionally substituted by substituent A, heterocyclyloxy optionally substituted by substituent A, carbocyclylamino optionally substituted by substituent A, or heterocyclylamino optionally substituted by substituent A;

the substituent A is selected from at least one of the group consisting of: oxo, alkyl, alkylamino, carbocyclyl, heterocyclyl, alkylcarbonyl, halogen, hydroxy, alkylcarbonylamino, alkylcarbonyloxy, alkoxycarbonyl, alkoxycarbonylalkyl, alkylaminocarbonyloxy, alkoxy, nitro, azido, alkylsulfonyl, and trialkylsilyl.

More specifically, in the compound represented by the formula (II), R₁And R₂Each independently hydrogen, halogen, alkyl, alkoxy; further preferably, R₁And R₂Each independently hydrogen, F, Cl, Br, C₁-C₄Alkyl radical, C₁-C₄An alkoxy group;

P^Ris composed of

Said C is₁-C₄Alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl. Said C is₁-C₄Alkoxy is as above C₁-C₄Alkoxy corresponding to alkyl.

According to the invention, in particular, the compounds of formula II are selected from:

(R) -12- (5H-dibenzo [ a, d ] [7] -5-cyclopentenyl) -7-hydroxy-3, 4,12,12 a-tetrahydro-1H- [1,4] oxazine [3,4-c ] pyrido [2,1-f ] [1,2,4] triazine-6, 8-dione (10 a);

(R) -7-hydroxy-12- (((R/S) -2-bromo-5H-dibenzo [ a, d ] [7] -5-cyclopentenyl) -7-hydroxy-3, 4,12,12 a-tetrahydro-1H- [1,4] oxazinyl [3,4-c ] pyridinyl [2,1-f ] [1,2,4] triazine-6, 8-dione (10 b);

(R) -7-hydroxy-12- (((R/S) -3-bromo-5H-dibenzo [ a, d ] [7] -5-cyclopentenyl) -7-hydroxy-3, 4,12,12 a-tetrahydro-1H- [1,4] oxazinyl [3,4-c ] pyridinyl [2,1-f ] [1,2,4] triazine-6, 8-dione (10 c);

(R) -7-hydroxy-12- (((R/S) -1-fluoro-5H-dibenzo [ a, d ] [7] -5-cyclopentenyl) -7-hydroxy-3, 4,12,12 a-tetrahydro-1H- [1,4] oxazinyl [3,4-c ] pyridinyl [2,1-f ] [1,2,4] triazine-6, 8-dione (10 d);

(R) -7-hydroxy-12- (((R/S) -2-fluoro-5H-dibenzo [ a, d ] [7] -5-cyclopentenyl) -7-hydroxy-3, 4,12,12 a-tetrahydro-1H- [1,4] oxazinyl [3,4-c ] pyridinyl [2,1-f ] [1,2,4] triazine-6, 8-dione (10 e);

(R) -7-hydroxy-12- (((R/S) -3-fluoro-5H-dibenzo [ a, d ] [7] -5-cyclopentenyl) -7-hydroxy-3, 4,12,12 a-tetrahydro-1H- [1,4] oxazinyl [3,4-c ] pyridinyl [2,1-f ] [1,2,4] triazine-6, 8-dione (10 f);

(R) -7-hydroxy-12- (((R/S) -1-chloro-5H-dibenzo [ a, d ] [7] -5-cyclopentenyl) -7-hydroxy-3, 4,12,12 a-tetrahydro-1H- [1,4] oxazinyl [3,4-c ] pyridinyl [2,1-f ] [1,2,4] triazine-6, 8-dione (10 g);

(R) -7-hydroxy-12- (((R/S) -2-chloro-5H-dibenzo [ a, d ] [7] -5-cyclopentenyl) -7-hydroxy-3, 4,12,12 a-tetrahydro-1H- [1,4] oxazinyl [3,4-c ] pyridinyl [2,1-f ] [1,2,4] triazine-6, 8-dione (10H);

(R) -7-hydroxy-12- (((R/S) -3-chloro-5H-dibenzo [ a, d ] [7] -5-cyclopentenyl) -7-hydroxy-3, 4,12,12 a-tetrahydro-1H- [1,4] oxazinyl [3,4-c ] pyridinyl [2,1-f ] [1,2,4] triazine-6, 8-dione (10 i);

(R) -7-hydroxy-12- (((R/S) -3-methyl-5H-dibenzo [ a, d ] [7] -5-cyclopentenyl) -3,4,12,12 a-tetrahydro-1H- [ [1,4] oxazinyl [3,4-c ] pyridinyl [2,1-f ] [1,2,4] triazine-6, 8-dione (10 j);

(R) -7-hydroxy-12- (((R/S) -3-methoxy-5H-dibenzo [ a, d ] [7] -5-cyclopentenyl) -3,4,12,12 a-tetrahydro-1H- [ [1,4] oxazinyl [3,4-c ] pyridinyl [2,1-f ] [1,2,4] triazine-6, 8-dione (10 k).

The invention provides a preparation method of the compound, which takes different substituted benzaldehydes as initial raw materials to obtain the compound shown in the general formula II through a plurality of reactions. The specific synthesis scheme is shown in FIG. 1.

The synthetic route shown in FIG. 1 outlines and describes the preparation of the compounds of formula II of the present invention, all starting materials are prepared by the methods described in these schemes, by methods well known to those of ordinary skill in the art of organic chemistry, or are commercially available. All of the final derivatives of the invention are prepared by the methods described in these schemes or by methods analogous thereto, which are well known to those of ordinary skill in the art of organic chemistry.

In a second aspect, the present invention provides a pharmaceutical composition comprising the polycyclic pyridone derivative as an active ingredient and a pharmaceutically acceptable excipient.

The invention can contain the derivatives of the compound shown in the general formula I and pharmaceutically acceptable salts and hydrates thereof as active ingredients, and the derivatives, the pharmaceutically acceptable salts and the hydrates are mixed with pharmaceutically acceptable excipients to prepare a composition and prepare a clinically acceptable dosage form, wherein the excipients refer to diluents, auxiliary agents or carriers which can be used in the pharmaceutical field. The above dosage forms are clinically common injections, tablets, capsules and the like.

The compound or the pharmaceutically acceptable salt, hydrate and prodrug thereof can be used alone as a sole anti-influenza medicament or can be combined with the anti-influenza medicaments on the market for preventing, treating or relieving symptoms of patients infected by influenza viruses.

In a third aspect, the present invention provides the use of the polycyclic pyridone derivative and/or the pharmaceutical composition for the preparation of a medicament for treating or preventing a disease caused by a virus having a cap-dependent endonuclease. Such viruses with cap-dependent endonucleases include, but are not limited to, influenza virus.

In a fourth aspect, the invention provides the use of the polycyclic pyridone derivative and/or the pharmaceutical composition in the preparation of an influenza virus RNA polymerase inhibitor.

In a fifth aspect, the present invention provides the use of a polycyclic pyridone derivative and/or a pharmaceutical composition as described above for the preparation of a medicament for preventing, treating or alleviating symptoms in a patient after an influenza virus infection.

The compound or the pharmaceutically acceptable salt, hydrate and prodrug thereof have remarkable inhibitory activity on influenza virus, wherein the activity of part of the compound is equivalent to that of baroxavir, even exceeds that of the baroxavir.

Additional features and advantages of the invention will be set forth in the detailed description which follows.

Drawings

The above and other objects, features and advantages of the present invention will become more apparent by describing in more detail exemplary embodiments thereof with reference to the attached drawings.

FIG. 1 is a scheme showing the synthesis scheme of the polycyclic pyridone derivatives of the present invention.

Detailed Description

Preferred embodiments of the present invention will be described in more detail below. While the following describes preferred embodiments of the present invention, it should be understood that the present invention may be practiced in various forms and should not be limited by the embodiments set forth herein.

In the following examples, methods of preparing the compounds are depicted. It is to be understood that the following methods, as well as other methods known to those of ordinary skill in the art, can be applied to the preparation of all of the compounds described herein. The examples are intended to illustrate, but not to limit, the scope of the invention.

Example 1: synthesis of (R) -12- (5H-dibenzo [ a, d ] [7] -5-cyclopentenyl) -7-hydroxy-3, 4,12,12 a-tetrahydro-1H- [1,4] oxazine [3,4-c ] pyrido [2,1-f ] [1,2,4] triazine-6, 8-dione (10a)

Step 1: a100 mL reaction flask was charged with 20mmol of compound 7a, 10mL of tetrahydrofuran and 2mL of anhydrous ethanol, 10mmol of sodium borohydride was slowly added under ice bath, and the mixture was stirred at room temperature for 1.5 hours. Pouring the reaction solution into ice water, decompressing and distilling off most of organic solvent, extracting the product by dichloromethane, drying the organic phase by anhydrous sodium sulfate, decompressing and distilling off the organic phase to obtain the compound 8a, and directly using the compound in the next reaction without purification.¹H NMR(400MHz,DMSO-d₆)δ7.68(dt,J＝7.8,1.1Hz,2H),7.39(td,J＝7.5,1.4Hz,2H),7.34(dd,J＝7.6,1.4Hz,2H),7.22(td,J＝7.4,1.4Hz,2H),7.13(s,2H),6.08(s,1H),4.99(d,J＝4.1Hz,1H).

Step 2: a100 mL reaction flask was charged with 1mmol of Compound 1, 1.2mmol of Compound 8a, and 2.5mmol of a 50 wt.% solution of 1-propylphosphoric anhydride in ethyl acetate and stirred at 50 deg.C overnight. Pouring the reaction solution into a proper amount of ice water, extracting the product with ethyl acetate, and washing the organic phase with saturated brineThe phases were dried over anhydrous sodium sulfate and evaporated to dryness under reduced pressure. The crude product was purified on a large silica gel plate (20 cm. times.20 cm). Compound 9a is obtained.¹H NMR(400MHz,Chloroform-d)δ7.65–7.57(m,2H),7.48(dt,J＝4.9,2.9Hz,1H),7.46–7.40(m,2H),7.40–7.33(m,4H),7.31–7.25(m,2H),7.04(d,J＝6.3Hz,3H),6.56(dd,J＝7.7,1.3Hz,1H),6.25(d,J＝7.7Hz,1H),5.68–5.53(m,2H),5.41(d,J＝10.8Hz,1H),5.33(s,1H),4.58(dd,J＝13.6,2.5Hz,1H),3.90(dd,J＝9.9,3.0Hz,1H),3.61(dd,J＝11.8,3.3Hz,1H),3.51(dd,J＝10.8,3.1Hz,1H),3.18(td,J＝11.8,2.7Hz,1H),3.12–3.01(m,1H),2.76(ddd,J＝13.5,11.8,3.5Hz,1H).

And step 3: a50 mL reaction flask was charged with 0.5mmol of compound 9a, 1.5mmol of lithium chloride and 5mL of DMA, and the reaction mixture was stirred at 80 ℃ for 3 hours. While stirring in ice bath, 2mL of acetone, 9mL of 0.5M diluted hydrochloric acid and 4mL of the reaction mixture were added, and stirring in ice bath was continued for 1 hour. The product is precipitated in solid form, and is filtered to obtain solid, and the solid is dried to obtain the target compound 10a which is light pink solid. The yield was 92.3%.¹H NMR(400MHz,Chloroform-d)δ7.55–7.39(m,5H),7.34(t,J＝7.5Hz,1H),7.19(t,J＝7.5Hz,1H),7.07(s,2H),6.91(d,J＝7.6Hz,1H),6.24(d,J＝7.7Hz,1H),5.58(d,J＝7.7Hz,1H),5.41(s,1H),4.57(dd,J＝13.5,2.6Hz,1H),4.03(dd,J＝10.0,3.1Hz,1H),3.70(dd,J＝12.0,3.4Hz,1H),3.61(dd,J＝11.0,3.1Hz,1H),3.40–3.23(m,2H),2.86(ddd,J＝15.1,11.8,3.5Hz,1H)；¹³C NMR(101MHz,Chloroform-d)δ172.23,162.04,154.02,137.85,134.51,134.40,133.08,133.05,130.89,130.73,130.69,130.38,130.33,130.18,129.55,129.34,129.29,114.79,110.93,77.28,75.72,69.51,69.04,66.54,45.59；HRMS-ESI(m/z)Calcd.For C₂₅H₂₂N₃O₄[M+H]⁺:428.1610,Found:428.1602.

Example 2: synthesis of (R) -7-hydroxy-12- (((R/S) -2-bromo-5H-dibenzo [ a, d ] [7] -5-cyclopentenyl) -7-hydroxy-3, 4,12,12 a-tetrahydro-1H- [1,4] oxazinyl [3,4-c ] pyridinyl [2,1-f ] [1,2,4] triazine-6, 8-dione (10b)

Step 1: a100 mL reaction flask was charged with Compound 2(3.15g,21.0mmol), NBS (4.09g,23.0mmol), and 30mL chlorobenzene in that order. The reaction mixture was heated to 70 ℃ and AIBN (0.13g,0.80mmol) was slowly added and stirring was continued at 70 ℃ for 1 hour. And (3) cooling the reaction liquid to room temperature, and evaporating the solvent in the reaction liquid under reduced pressure to obtain a crude product of the compound 3, wherein the crude product is directly used for the next reaction without purification.

Step 2: a100 mL reaction flask was charged with the compound 3 obtained in the previous step, triphenylphosphine (5.51g, 21.0mmol) and 25mL acetone in that order, and the reaction was refluxed for 1 hour. And (4) after the reaction liquid is cooled to room temperature, carrying out suction filtration to obtain a white solid. A100 mL reaction flask was charged with the white solid obtained above, sodium methoxide (1.35g, 25.0mmol), and 30mL of methanol. The reaction mixture was stirred at room temperature for half an hour, then heated under reflux, and 11mmol 4b was added thereto and refluxed for 6 hours. And (3) cooling the reaction liquid to room temperature, pouring the reaction liquid into a large amount of ice water, extracting a product by using dichloromethane, drying an organic phase by using anhydrous sodium sulfate, and evaporating to dryness under reduced pressure to obtain a crude product. And mixing the crude product with 50mL of petroleum ether and performing ultrasonic treatment, separating out most impurities in a solid form, filtering out the solid, and evaporating the filtrate under reduced pressure to obtain a primarily purified compound 5b, wherein the compound is directly used for the next reaction without further purification.¹H NMR(400MHz,Chloroform-d)δ8.01(m,1H),7.34–7.32(m,2H),7.25–7.13(m,3H),7.11(d,J＝12.0Hz,1H),6.99–6.92(m,2H),6.57(d,J＝12.0Hz,1H),3.90(s,3H).

And step 3: a100 mL reaction flask was charged with 5.0mmol 5b of the compound obtained in the previous step, potassium hydroxide (0.56g,10.0mmol), 20mL methanol and 1mL water, and the reaction solution was refluxed for 4 hours. And cooling the reaction liquid to room temperature, pouring the reaction liquid into a large amount of ice water, stirring continuously, adding a 4M hydrochloric acid solution for acidification, and precipitating a crude product in a solid form. And (3) carrying out suction filtration to obtain a solid, mixing the dried solid with 50mL of petroleum ether, carrying out ultrasonic treatment, dissolving most of impurities in the petroleum ether, carrying out suction filtration, washing the solid with the petroleum ether to obtain a primarily purified compound 6b, and directly using the primarily purified compound in the next reaction without continuous purification.¹H NMR(400MHz,Chloroform-d)δ8.13-8.11(m,1H),7.38–7.34(m,2H),7.24–7.18(m,3H),7.14(d,J＝12.0Hz,1H).7.01–6.94(m,2H),6.59(d,J＝12.0Hz,1H).

And 4, step 4: a100 mL reaction flask was charged with 1.0mmol 6b of the compound obtained in the previous step, dissolved in 20mL of dichloromethane, and thionyl chloride (0.13g,1.1mmol) was slowly added thereto with stirring at room temperature, and then the reaction solution was refluxed for 1.5 hours. To be treatedThe reaction mixture was cooled to room temperature, and 0.17g of anhydrous aluminum chloride was slowly added thereto, followed by stirring at room temperature for 3 hours. Pouring the reaction liquid into ice water, extracting a product by using dichloromethane, drying an organic phase by using anhydrous sodium sulfate, and evaporating the organic phase by reduced pressure to obtain a crude product of the compound 7b, wherein the crude product is directly used for the next reaction without purification.¹H NMR(400MHz,Chloroform-d)δ8.26–8.22(m,1H),8.11(d,J＝8.5Hz,1H),7.72(d,J＝2.0Hz,1H),7.71–7.64(m,2H),7.62–7.55(m,2H),7.12(d,J＝12.1Hz,1H),6.97(d,J＝12.1Hz,1H).

And 5: the same procedure as in step 1 of example 1 gave compound 8 b.¹H NMR(400MHz,Chloroform-d)δ7.69(d,J＝7.8Hz,1H),7.58(d,J＝8.3Hz,1H),7.56–7.49(m,2H),7.46(td,J＝7.5,1.5Hz,1H),7.38(dd,J＝7.7,1.4Hz,1H),7.32(dd,J＝7.4,1.3Hz,1H),7.17(d,J＝11.7Hz,1H),7.03(d,J＝11.7Hz,1H),5.37(s,1H).

Step 6: the same procedure as in step 2 of example 1 gave compound 9 b. A pair of diastereomers in a ratio of 3: 4.¹H NMR(400MHz,Chloroform-d)δ7.68–7.63(m,4H),7.62–7.59(m,3H),7.55–7.52(m,3H),7.48–7.45(m,4H),7.38–7.35(m,4H),7.34–7.31(m,2H),7.16–7.02(m,4H),6.93(dd,J＝11.8,1.5Hz,2H),6.59–6.18(m,4H),5.77–5.55(m,4H),5.43(t,J＝11.4Hz,2H),5.29(d,J＝13.2Hz,2H),4.59(ddd,J＝13.5,7.1,2.5Hz,2H),3.87(ddd,J＝25.7,9.9,3.0Hz,2H),3.63(td,J＝11.5,3.3Hz,2H),3.53(ddd,J＝10.0,6.5,3.0Hz,2H),3.26–3.13(m,2H),3.07(t,J＝10.4Hz,2H),2.76(dddd,J＝25.1,13.5,11.7,3.5Hz,2H).

And 7: the same procedure as in step 3 of example 1 gave compound 10 b. The yield was 91.5%. Diastereoisomers, ratio 3: 7.¹H NMR(400MHz,Chloroform-d)δ7.69–7.30(m,5H),7.21(t,J＝7.6Hz,1H),7.11(t,J＝8.9Hz,1H),7.05–6.72(m,2H),6.25(dd,J＝25.7,7.6Hz,1H),5.63(dd,J＝47.5,7.7Hz,1H),5.38(s,1H),4.57(dd,J＝13.2,8.6Hz,1H),4.01(ddd,J＝23.2,10.1,3.0Hz,1H),3.66(dtd,J＝36.2,9.3,6.7,3.2Hz,2H),3.32(dtd,J＝20.8,11.3,10.4,3.4Hz,2H),2.98–2.75(m,1H)；¹³C NMR(101MHz,Chloroform-d)δ172.19,162.03,154.03,137.77,136.16,133.44,132.76,132.21,132.07,131.90,131.84,131.76,131.09,130.72,130.41,130.32,129.97,129.71,129.59,129.17,128.83,123.33,114.72,111.16,110.96,75.09,74.92,69.61,69.50,69.00,66.53,45.66；HRMS-ESI(m/z)Calcd.For C₂₅H₂₁BrN₃O₄[M+H]⁺:506.0715,Found:506.0708.

Example 3: synthesis of (R) -7-hydroxy-12- (((R/S) -3-bromo-5H-dibenzo [ a, d ] [7] -5-cyclopentenyl) -7-hydroxy-3, 4,12,12 a-tetrahydro-1H- [1,4] oxazinyl [3,4-c ] pyridinyl [2,1-f ] [1,2,4] triazine-6, 8-dione (10c)

The method comprises the following steps: the same procedure as in steps 1-7 of example 2 gave compound 10 c. The yield was 93.6%. Diastereoisomers in a ratio of 1: 1.¹H NMR(400MHz,DMSO-d₆)δ11.70(s,2H),7.99(d,J＝2.1Hz,1H),7.69(dd,J＝8.3,2.1Hz,1H),7.62(dq,J＝8.0,3.9Hz,2H),7.59–7.54(m,2H),7.54–7.47(m,4H),7.44(d,J＝8.3Hz,1H),7.38(td,J＝7.5,1.4Hz,1H),7.27–7.10(m,6H),6.35(dd,J＝15.6,7.7Hz,2H),5.85(d,J＝2.8Hz,2H),5.43(d,J＝7.7Hz,1H),5.34(d,J＝7.6Hz,1H),4.40(ddd,J＝16.2,13.4,2.5Hz,2H),3.85(dd,J＝8.4,4.6Hz,1H),3.76(dd,J＝9.5,3.5Hz,1H),3.59(ddd,J＝14.4,11.6,3.3Hz,2H),3.41(td,J＝10.0,9.0,3.6Hz,4H),3.29(dd,J＝11.3,2.7Hz,2H),2.84(ddd,J＝14.7,11.8,3.5Hz,1H),2.75(ddd,J＝15.2,11.1,3.4Hz,1H)；¹³C NMR(101MHz,DMSO-d₆)δ171.39,161.71,153.44,138.58,135.93,135.03,133.86,133.67,133.62,133.48,133.39,132.73,132.29,132.14,131.84,131.57,131.42,131.26,131.18,130.95,130.50,129.86,129.71,129.62,129.40,122.67,122.26,116.00,115.93,110.36,110.33,72.63,72.52,69.94,69.69,68.22,65.94,45.72；HRMS-ESI(m/z)Calcd.For C₂₅H₂₁BrN₃O₄[M+H]⁺:506.0715,Found:506.0709.

Example 4: synthesis of (R) -7-hydroxy-12- (((R/S) -1-fluoro-5H-dibenzo [ a, d ] [7] -5-cyclopentenyl) -7-hydroxy-3, 4,12,12 a-tetrahydro-1H- [1,4] oxazinyl [3,4-c ] pyridinyl [2,1-f ] [1,2,4] triazine-6, 8-dione (10d)

The method comprises the following steps: the same procedure as in steps 1-7 of example 2 gave compound 10 d. The yield was 92.1%. Diastereoisomers in a ratio of 1: 1.¹H NMR(400MHz,Chloroform-d)δ7.60–7.27(m,9H),7.18(ddt,J＝22.0,15.9,8.2Hz,6H),7.06(t,J＝9.1Hz,1H),6.90(d,J＝7.6Hz,1H),6.71(d,J＝7.6Hz,1H),6.33(d,J＝7.7Hz,1H),6.25(d,J＝7.7Hz,1H),5.66(d,J＝7.7Hz,1H),5.57(d,J＝7.7Hz,1H),5.44(s,2H),4.57(ddd,J＝13.4,6.0,2.5Hz,2H),4.04(ddd,J＝22.5,10.0,3.1Hz,2H),3.67(dtd,J＝35.1,11.5,3.3Hz,4H),3.32(dtd,J＝20.8,10.4,9.4,3.5Hz,4H),2.86(qd,J＝13.0,3.6Hz,2H)；¹³C NMR(101MHz,Chloroform-d)δ172.20,162.00,154.07,154.03,137.78,137.72,134.31,134.20,133.00,132.93,131.91,131.62,131.04,130.71,130.56,130.41,130.34,129.88,129.64,129.57,126.17,125.77,121.91,121.46,121.38,116.33,116.19,116.10,114.75,114.69,111.09,110.97,75.10,75.04,69.69,69.54,69.01,68.98,66.53,45.61；HRMS-ESI(m/z)Calcd.For C₂₅H₂₁FN₃O₄[M+H]⁺:446.1516,Found:446.1507.

Example 5: synthesis of (R) -7-hydroxy-12- (((R/S) -2-fluoro-5H-dibenzo [ a, d ] [7] -5-cyclopentenyl) -7-hydroxy-3, 4,12,12 a-tetrahydro-1H- [1,4] oxazinyl [3,4-c ] pyridinyl [2,1-f ] [1,2,4] triazine-6, 8-dione (10e)

The method comprises the following steps: the same procedure as in steps 1-7 of example 2 gave compound 10 e. The yield was 91.0%. Diastereoisomers in a ratio of 1: 1.¹H NMR(400MHz,Chloroform-d)δ7.58–7.47(m,3H),7.43(td,J＝8.7,8.0,4.9Hz,3H),7.37(td,J＝7.6,1.3Hz,1H),7.23(ddd,J＝9.2,6.4,1.8Hz,2H),7.19–7.08(m,4H),7.00(dd,J＝11.8,4.7Hz,2H),6.91(dd,J＝7.2,1.9Hz,3H),6.29(d,J＝7.7Hz,1H),6.23(d,J＝7.7Hz,1H),5.69(d,J＝7.7Hz,1H),5.59(d,J＝7.7Hz,1H),5.41(s,2H),4.59(ddd,J＝13.5,5.3,2.5Hz,2H),4.03(ddd,J＝11.1,10.0,3.1Hz,2H),3.72(ddd,J＝11.5,7.7,3.4Hz,2H),3.63(ddd,J＝10.8,7.5,3.1Hz,2H),3.32(dddd,J＝23.3,11.3,9.4,2.6Hz,4H),2.87(dtd,J＝15.4,12.1,3.5Hz,2H)；¹³C NMR(101MHz,Chloroform-d)δ172.24,162.07,154.07,137.78,136.47,134.14,132.53,131.93,131.59,131.09,130.68,130.40,130.31,129.97,129.68,129.51,129.11,117.15,116.94,116.35,111.03,110.96,74.91,74.81,69.51,69.05,66.55,45.66,45.60；HRMS-ESI(m/z)Calcd.For C₂₅H₂₁FN₃O₄[M+H]⁺:446.1516,Found:446.1506.

Example 6: synthesis of (R) -7-hydroxy-12- (((R/S) -3-fluoro-5H-dibenzo [ a, d ] [7] -5-cyclopentenyl) -7-hydroxy-3, 4,12,12 a-tetrahydro-1H- [1,4] oxazinyl [3,4-c ] pyridinyl [2,1-f ] [1,2,4] triazine-6, 8-dione (10f)

The method comprises the following steps: the same procedure as in steps 1-7 of example 2 gave compound 10 f. The yield was 93.3%. Diastereoisomers in a ratio of 1: 1.¹H NMR(400MHz,Chloroform-d)δ7.58–7.30(m,8H),7.19(q,J＝8.6,7.8Hz,3H),7.05(s,5H),6.90(d,J＝7.7Hz,1H),6.69(d,J＝8.6Hz,1H),6.28(dd,J＝29.0,7.7Hz,2H),5.62(dd,J＝39.0,7.7Hz,2H),5.36(d,J＝4.6Hz,2H),4.59(t,J＝14.8Hz,2H),4.03(ddd,J＝23.6,9.9,3.1Hz,2H),3.83–3.53(m,4H),3.50–3.13(m,4H),3.03–2.76(m,2H)；¹³C NMR(101MHz,Chloroform-d)δ172.21,162.06,161.99,154.05,137.79,134.43,134.25,132.86,132.78,132.41,132.29,132.03,130.89,130.76,130.31,130.19,129.87,129.69,129.58,129.46,129.21,117.45,116.94,116.71,111.04,110.95,75.03,69.70,69.49,69.03,68.93,66.55,45.69,45.65.；HRMS-ESI(m/z)Calcd.For C₂₅H₂₁FN₃O₄[M+H]⁺:446.1516,Found:446.1506.

Example 7: synthesis of (R) -7-hydroxy-12- (((R/S) -1-chloro-5H-dibenzo [ a, d ] [7] -5-cyclopentenyl) -7-hydroxy-3, 4,12,12 a-tetrahydro-1H- [1,4] oxazinyl [3,4-c ] pyridinyl [2,1-f ] [1,2,4] triazine-6, 8-dione (10g)

The method comprises the following steps: the same procedures in steps 1 to 7 of example 2 gave 10g of a compound.¹H NMR(400MHz,Chloroform-d)δ7.56–7.33(m,12H),7.25–7.16(m,3H),7.10(t,J＝7.8Hz,1H),6.90(d,J＝7.6Hz,1H),6.83(d,J＝7.6Hz,1H),6.33(d,J＝7.6Hz,1H),6.26(d,J＝7.7Hz,1H),5.68(d,J＝7.7Hz,1H),5.59(d,J＝7.7Hz,1H),5.40(s,2H),4.57(ddd,J＝13.3,7.1,2.5Hz,2H),4.08(dd,J＝9.9,3.1Hz,1H),4.01(dd,J＝10.0,3.1Hz,1H),3.78–3.58(m,4H),3.41–3.23(m,4H),2.86(dddd,J＝15.3,13.4,11.8,3.5Hz,2H)；¹³C NMR(101MHz,Chloroform-d)δ172.21,161.94,154.11,154.05,137.76,135.76,135.61,133.90,133.32,131.87,131.61,130.90,130.83,130.77,130.47,130.15,130.11,129.97,129.92,129.78,129.70,129.64,129.23,128.89,126.42,126.05,114.70,111.13,111.01,77.28,75.39,75.25,69.79,69.59,69.00,68.98,66.54,45.62,45.59；HRMS-ESI(m/z)Calcd.For C₂₅H₂₁ClN₃O₄[M+H]⁺:462.1221,Found:462.1211.

Example 8: synthesis of (R) -7-hydroxy-12- (((R/S) -2-chloro-5H-dibenzo [ a, d ] [7] -5-cyclopentenyl) -7-hydroxy-3, 4,12,12 a-tetrahydro-1H- [1,4] oxazinyl [3,4-c ] pyridinyl [2,1-f ] [1,2,4] triazine-6, 8-dione (10H)

The method comprises the following steps: the same procedure as in steps 1-7 of example 2 gave compound 10 h. The yield was 92.5%. Diastereoisomers in a ratio of 1: 1.¹H NMR(400MHz,Chloroform-d)δ7.57–7.30(m,11H),7.24–7.07(m,4H),6.98(dd,J＝11.8,5.5Hz,2H),6.87(dd,J＝16.6,7.9Hz,2H),6.29(d,J＝7.7Hz,1H),6.22(d,J＝7.7Hz,1H),5.68(d,J＝7.7Hz,1H),5.57(d,J＝7.6Hz,1H),5.39(s,2H),4.57(ddd,J＝13.7,7.1,2.4Hz,2H),4.01(ddd,J＝19.2,9.9,3.1Hz,2H),3.71(td,J＝11.3,3.4Hz,2H),3.61(ddd,J＝10.0,6.5,3.1Hz,2H),3.42–3.22(m,4H),2.95–2.76(m,2H)；¹³C NMR(101MHz,Chloroform-d)δ172.19,162.02,154.02,137.79,135.89,135.25,134.19,132.89,132.07,131.90,131.73,131.69,131.09,130.70,130.48,130.41,130.33,129.94,129.78,129.71,129.58,129.27,129.14,128.91,114.75,111.14,110.96,77.28,75.00,74.86,69.59,69.51,68.99,66.52,45.67,45.62；HRMS-ESI(m/z)Calcd.For C₂₅H₂₁ClN₃O₄[M+H]⁺:462.1221,Found:462.1208.

Example 9: synthesis of (R) -7-hydroxy-12- (((R/S) -3-chloro-5H-dibenzo [ a, d ] [7] -5-cyclopentenyl) -7-hydroxy-3, 4,12,12 a-tetrahydro-1H- [1,4] oxazinyl [3,4-c ] pyridinyl [2,1-f ] [1,2,4] triazine-6, 8-dione (10i)

The method comprises the following steps: the same procedure as in steps 1-7 of example 2 gave compound 10 i. The yield was 90.7%. Diastereoisomers in a ratio of 1: 1.¹H NMR(400MHz,Chloroform-d)δ7.56–7.29(m,11H),7.22(td,J＝7.5,1.4Hz,1H),7.13–6.99(m,4H),6.97(d,J＝2.1Hz,1H),6.94–6.88(m,1H),6.31(d,J＝7.7Hz,1H),6.24(d,J＝7.7Hz,1H),5.69(d,J＝7.7Hz,1H),5.58(d,J＝7.7Hz,1H),5.36(d,J＝2.8Hz,2H),4.60(ddd,J＝19.8,13.3,2.5Hz,2H),4.03(ddd,J＝23.8,9.9,3.1Hz,2H),3.73(ddd,J＝16.3,12.0,3.4Hz,2H),3.62(ddd,J＝10.9,9.3,3.1Hz,2H),3.42–3.23(m,4H),2.94(ddd,J＝13.4,11.7,3.5Hz,1H),2.84(ddd,J＝13.4,11.8,3.5Hz,1H)；¹³C NMR(101MHz,Chloroform-d)δ172.22,162.05,154.10,137.81,137.75,135.55,134.40,132.99,132.90,132.64,132.49,132.12,131.27,131.09,130.97,130.80,130.34,130.29,130.01,129.86,129.65,129.59,129.53,129.45,129.36,129.23,114.67,114.64,110.95,77.25,75.08,74.92,69.68,69.48,69.04,68.95,66.56,66.51,45.68,45.63,29.71；HRMS-ESI(m/z)Calcd.For C₂₅H₂₁ClN₃O₄[M+H]⁺:462.1221,Found:462.1211.

Example 10: synthesis of (R) -7-hydroxy-12- (((R/S) -3-methyl-5H-dibenzo [ a, d ] [7] -5-cyclopentenyl) -3,4,12,12 a-tetrahydro-1H- [ [1,4] oxazinyl [3,4-c ] pyridinyl [2,1-f ] [1,2,4] triazine-6, 8-dione (10j)

The method comprises the following steps: the same procedure as in steps 1 to 7 of example 2 gave compound 10 j. The yield was 85.3%. Diastereoisomers in a ratio of 1: 1.¹H NMR(400MHz,Chloroform-d)δ7.53–7.38(m,6H),7.35–7.27(m,3H),7.23(d,J＝1.8Hz,1H),7.16(ddd,J＝12.8,7.6,1.5Hz,2H),7.07–6.98(m,4H),6.88(d,J＝7.6Hz,1H),6.72(d,J＝1.7Hz,1H),6.25(dd,J＝7.6,4.9Hz,2H),5.67–5.46(m,2H),5.35(s,2H),4.58(ddd,J＝13.5,7.6,2.5Hz,2H),4.05(ddd,J＝16.5,9.9,3.1Hz,2H),3.70(dt,J＝11.9,4.2Hz,2H),3.65–3.54(m,2H),3.50–3.13(m,4H),2.86(ddt,J＝12.3,8.7,2.8Hz,2H),2.41(s,3H),2.20(s,3H)；¹³C NMR(101MHz,Chloroform-d)δ172.20,162.01,153.97,140.05,139.67,138.03,137.90,134.58,132.94,132.87,131.92,131.30,130.89,130.82,130.80,130.67,130.63,130.34,130.26,130.21,130.16,130.10,130.06,129.78,129.44,129.33,129.27,129.25,129.07,114.87,114.83,110.91,110.76,75.80,69.48,69.04,66.52,45.57,21.13,20.96；HRMS-ESI(m/z)Calcd.For C₂₆H₂₄N₃O₄[M+H]⁺:442.1767,Found:442.1758.

Example 11: synthesis of (R) -7-hydroxy-12- (((R/S) -3-methoxy-5H-dibenzo [ a, d ] [7] -5-cyclopentenyl) -3,4,12,12 a-tetrahydro-1H- [ [1,4] oxazinyl [3,4-c ] pyridinyl [2,1-f ] [1,2,4] triazine-6, 8-dione (10k)

The method comprises the following steps: the same procedure as in steps 1-7 of example 2 gave compound 10 k. The yield was 94.9%. Diastereoisomers in a ratio of 1: 1.¹H NMR(400MHz,Chloroform-d)δ7.70–7.31(m,6H),7.08–6.84(m,3H),6.43(s,1H),6.31(d,J＝7.7Hz,1H),5.65(d,J＝7.7Hz,1H),5.34(s,1H),4.58(d,J＝12.9Hz,1H),4.03(dd,J＝9.8,3.0Hz,1H),3.85(d,J＝6.3Hz,1H),3.66(s,3H),3.32(dt,J＝21.7,11.1Hz,2H),2.86(t,J＝12.5Hz,1H)；¹³C NMR(101MHz,Chloroform-d)δ172.15,160.93,153.97,138.04,132.37,131.76,130.74,130.37,129.91,129.31,128.91,128.46,115.61,115.34,110.92,75.85,69.44,69.03,66.54,55.61,45.58,29.71；HRMS-ESI(m/z)Calcd.For C₂₆H₂₄N₃O₅[M+H]⁺:458.1716,Found:458.1707.

Biological Activity Studies of Compounds

In the following examples, some of the compounds of the present invention were used as examples to examine the antiviral activity and cytotoxicity of the compounds of the present invention.

Antiviral Activity and cytotoxicity assays

Example A: cytopathic effect experiment (CPE assay):

the ability of the compounds to inhibit the Cytopathic (CPE) effects of the viruses HlNl A/FortMonmouth/1/1947 and H3N 2A/WuhanHanFang/95/359 was tested at the cellular level in vitro.

The experimental steps are as follows:

(1) MDCK cells with 2.5 × 10⁴Density per well was seeded into 96-well plates.

(2) After 24h, the cells were washed once with PBS and infected with 100-half of the tissue cells (50% tissue culture infectious diseases, TCID)₅₀) The virus-infected solution (serum-free MEM medium) of (a) infected MDCK cells. Influenza virus: A/FortMonmouth/1/1947(H1N1), A/WuhanHanFang/359/1995(H3N 2).

(3) After 2h of infection at 37 ℃ unadsorbed virus was removed and a virus maintenance solution containing compounds was added (MEM medium supplemented with 2. mu.g/mL of TPCK treated pancreatin and 0.08% BSA). The highest concentration of compound detected was 200nM, 3-fold dilution, 8 concentrations, in order: 100nM, 66.67nM, 22.22nM, 7.41nM, 2.47nM, 0.82nM, 0.27nM, 0.09 nM. And a virus control group without medicine and a cell control group without medicine and virus infection are arranged at the same time. The cytotoxicity test group did not add virus and was replaced with medium. Two complex holes are arranged.

(4) Incubation is continued in the incubator at 37 ℃ for about 2 days, and the CPE result of the compound group is read when the cytopathic effect (CPE) degree of the virus control group is 75-100%.

(5) By Reed&The Muench method calculates the half effective Inhibitory Concentration (IC) of the drug₅₀) And according to TC₅₀/IC₅₀The ratio was used to calculate the therapeutic Index (Selective Index, SI).

The antiviral activity test and the cytotoxicity test show that the compound has good antiviral activity, and particularly, the inhibitory activity of part of the compound on influenza virus (A/FortMonmouth/1/1947) and influenza virus (A/WuhanhanHanFang/359/1995) is equivalent to that of baroxavir; at the same time, the compounds of the invention have very low cytotoxicity, TC₅₀Much less than the cytotoxicity of baroxavir.

TABLE 1 in vitro assay TC for influenza Virus (A/FortMonmouth/1/1947, A/WuhanHanFang/359/1995) with the compounds of the invention₅₀And IC₅₀Activity data

Note: TC (tungsten carbide)₅₀: half toxic concentration of drug; IC (integrated circuit)₅₀: the half inhibitory concentration of the drug on the virus.

As shown in Table 1, the compound of the present invention has excellent anti-influenza virus activity and low cytotoxicity.

Having described embodiments of the present invention, the foregoing description is intended to be exemplary, not exhaustive, and not limited to the embodiments disclosed. Many modifications and variations will be apparent to those of ordinary skill in the art without departing from the scope and spirit of the described embodiments.

Claims

1. A polycyclic pyridone derivative which is a compound of formula (I) or a stereoisomer, a tautomer, a nitrogen oxide, a solvate, a metabolite, a pharmaceutically acceptable salt of a compound of formula (I), or a prodrug thereof;

wherein,

p is hydrogen, benzyl, n-hexyl or P forming a prodrug^RA group;

A₁is CR_1AR_1BS or O;

A₂is CR_2AR_2BS or O;

A₃is CR_3AR_3BS or O;

A₄each independently is CR_4AR_4BS or O;

R_3Aand R_3BOptionally together with adjacent carbon atoms forming a carbocyclic ring orA heterocycle;

n is 1 or 2;

P^Ris at least one of the groups of formulae a) to ac) selected from:

a)-C(＝O)-P^R0；

b)-C(＝O)-P^R1；

c)-C(＝O)-L-P^R1；

d)-C(＝O)-L-O-P^R1；

e)-C(＝O)-L-O-L-O-P^R1；

f)-C(＝O)-L-O-C(＝O)-O-P^R1；

g)-C(＝O)-O-P^R2；

h)-C(＝O)-N(-K)(P^R2)；

i)-C(＝O)-O-L-O-P^R2；

j)-C(P^R3)₂-O-P^R4；

k)-C(P^R3)₂-O-L-O-P^R4；

l)-C(P^R3)₂-O-C(＝O)-P^R4；

m)-C(P^R3)₂-O-C(＝O)-O-P^R4；

n)-C(P^R3)₂-O-C(＝O)-N(-K)-P^R4；

o)-C(P^R3)₂-O-C(＝O)-O-L-O-P^R4；

p)-C(P^R3)₂-O-C(＝O)-O-L-N(P^R4)₂；

q)-C(P^R3)₂-O-C(＝O)-N(-K)-L-O-P^R4；

r)-C(P^R3)₂-O-C(＝O)-N(-K)-L-N(P^R4)₂；

s)-C(P^R3)₂-O-C(＝O)-O-L-O-L-O-P^R4；

t)-C(P^R3)₂-O-C(＝O)-O-L-N(-K)-C(＝O)-P^R4；

u)-C(P^R3)₂-O-P(＝O)(-P^R5)₂；

v)-C(P^R3)₂-P^R6)₂；

w)-C(＝N⁺(P^R7)₂)(-N(P^R7)₂)；

x)-C(P^R3)₂-C(P^R3)₂-C(C＝O)-O-P^R2；

y)-C(P^R3)₂-N(-K)-C(C＝O)-O-P^R2；

z)-P(＝O)(-P^R8)(-P^R9)；

aa)-S(＝O)₂-P^R10；

ab)-P^R11；

ac)-C(P^R3)₂-C(P^R3)₂-O-P^R2；

k is hydrogen or alkyl optionally substituted by substituent group a;

P^R3each independently is hydrogen, alkyl;

P^R4each independently is alkyl optionally substituted with substituent A, carbocyclyl optionally substituted with substituent A, heterocyclyl optionally substituted with substituent A, alkylamino optionally substituted with substituent A, carbocycloalkyl optionally substituted with substituent A, optionally substitutedHeterocycloalkyl substituted with group a, or trialkylsilyl;

P^R5each independently is OBn;

P^R7each independently is alkyl optionally substituted with substituent A;

P^R8is alkoxy optionally substituted with substituent a;

2. A polycyclic pyridone derivative according to claim 1, wherein the polycyclic pyridone derivative is a compound represented by formula (II) or a stereoisomer, a tautomer, a nitrogen oxide, a solvate, a metabolite, a pharmaceutically acceptable salt thereof, or a prodrug thereof,

wherein,

p is hydrogen, benzyl, n-hexyl or P forming a prodrug^RA group;

P^Ris at least one selected from the following groups:

a)-C(＝O)-P^R0；

b)-C(＝O)-P^R1；

g)-C(＝O)-O-P^R2；

h)-C(＝O)-N(-K)(P^R2)；

i)-C(＝O)-O-L-O-P^R2；

l)-C(P^R3)₂-O-C(＝O)-P^R4；

m)-C(P^R3)₂-O-C(＝O)-O-P^R4；

o)-C(P^R3)₂-O-C(＝O)-O-L-O-P^R4；

v)-C(P^R3)₂-P^R6)₂；

x)-C(P^R3)₂-C(P^R3)₂-C(C＝O)-O-P^R2；

y)-C(P^R3)₂-N(-K)-C(C＝O)-O-P^R2；

z)-P(＝O)(-P^R8)(-P^R9)；

wherein L is a linear or branched alkylene group;

k is hydrogen or alkyl optionally substituted with substituent A;

P^R0is alkyl optionally substituted with substituent A;

P^R3each independently is hydrogen, alkyl;

P^R8is alkoxy optionally substituted with substituent a;

3. A polycyclic pyridone derivative according to claim 2, wherein,

R₁and R₂Each independently hydrogen, halogen, alkyl, alkoxy;

P^Ris composed of

4. A polycyclic pyridone derivative according to claim 3, wherein,

R₁and R₂Each independently hydrogen, F, Cl, Br, C₁-C₄Alkyl radical, C₁-C₄An alkoxy group.

5. A polycyclic pyridone derivative according to claim 1, wherein the compound of formula II is selected from:

6. A pharmaceutical composition comprising a polycyclic pyridone derivative according to any one of claims 1 to 5 as an active ingredient and a pharmaceutically acceptable excipient.

7. The pharmaceutical composition of claim 6, wherein the pharmaceutical composition further comprises an additional anti-influenza drug.

8. Use of a polycyclic pyridone derivative according to any one of claims 1 to 5 and/or a pharmaceutical composition according to any one of claims 6 to 7 for the preparation of an influenza virus RNA polymerase inhibitor.

9. Use of a polycyclic pyridone derivative according to any one of claims 1 to 5 and/or a pharmaceutical composition according to any one of claims 6 to 7 for the preparation of a medicament for the treatment or prevention of a disease caused by a virus having a cap-dependent endonuclease; the virus having a cap-dependent endonuclease is preferably an influenza virus.

10. Use of a polycyclic pyridone derivative according to any one of claims 1 to 5 and/or a pharmaceutical composition according to any one of claims 6 to 7 for the manufacture of a medicament for preventing, treating or alleviating symptoms in a patient after an influenza virus infection.