CN111606928A

CN111606928A - Azacyclodiketone compound and preparation method thereof

Info

Publication number: CN111606928A
Application number: CN201910132596.3A
Authority: CN
Inventors: 沈旺; 党奎峰; 刘劲强; 蔡雨婷; 陈春明; 韩巧; 靡佳晨; 沈家磊; 孙亚波; 王鑫; 张超; 马随军; 赖新忠
Original assignee: Weiqing Biotechnology Shanghai Co ltd
Current assignee: Weiqing Biotechnology Shanghai Co ltd
Priority date: 2019-02-22
Filing date: 2019-02-22
Publication date: 2020-09-01

Abstract

The invention provides a nitrogen heterocyclic diketone compound which is characterized by being a compound shown in the following structure:

Description

Azacyclodiketone compound and preparation method thereof

Technical Field

The invention relates to a compound with inhibitory activity on influenza virus proliferation, in particular to a compound which can inhibit influenza virus proliferation by inhibiting cap dependent endonuclease related to influenza so as to treat or prevent influenza. In particular, substituted polycyclic pyridopyridazinedione derivatives having inhibitory activity against cap-dependent endonucleases, and pharmaceutical compositions containing them.

Background

Influenza viruses belong to the Orthomyxoviridae (Orthomyxoviridae) family of viruses, which are negative-sense single-stranded RNA viruses. Three influenza viruses are known: influenza virus a, influenza virus B, influenza virus C. Infection with influenza virus causes acute respiratory infections and can cause other respiratory viral infections, including upper and lower respiratory viral infections. Viral infections of the upper respiratory tract involve the nose, sinuses, pharynx and/or larynx. Viral infections of the lower respiratory tract involve the respiratory system below the vocal cords, including the trachea, main/bronchi and lungs. Infection with influenza virus causes millions of deaths per year. Influenza is a particularly important disease in high risk populations such as infants, the elderly, and the like. The pneumonia complication rate caused by the influenza of the old people is high, and the number of the old people dead due to the influenza accounts for the majority.

Influenza infection in humans is caused by influenza subtypes a and B. Influenza A virus infection accounts for the majority of infections, and patients are more seriously ill than influenza B virus infection. Influenza a virus has a host cell-derived lipid membrane that contains hemagglutinin, neuraminidase, and M2 proteins protruding from the surface of the virus. Influenza a viruses can be further classified according to the hemagglutinin (H or HA) and neuraminidase (N) antigen types. 16H antigens (H1 to H16) and 9N antigens (NI to N9) are now known. Influenza a viruses are divided into several subtypes based on the binding of H and N antigens. Subtypes that have now been found to exist include H1N1, H1N2, H2N2, H3N1, H3N2, H3N8, H5N1, H5N2, H5N3, H5N8, H5N9, H7N1, H7N2, H7N3, H7N4, H7N7, H7N9, H9N2 and H10N 7. Influenza polymerase is a heterotrimer composed of 3 subunits: polymerase Acid (PA), polymerase base 1(PB1), and polymerase base 2(PB 2). In the nucleus of infected cells, influenza virus polymerase is responsible for the replication and transcription of viral RNA. The PA subunit comprises an endonuclease active site. The endonuclease of PA cleaves mRNA of the host cell, which is then used by the PB1 subunit as a primer for viral mRNA synthesis.

Influenza viruses can be transmitted from person to person by direct contact with secretions of influenza patients, or with surfaces or objects that are contaminated. Complications of influenza virus infection include pneumonia, bronchitis, dehydration, and sinus and ear infections.

Anti-influenza drugs that have been approved fall into two broad categories: (1) the drugs for inhibiting the uncoating process of viruses include, but are not limited to, famethrel (Symmetrel, trade name: Amantadine, and Rimantadine hydrochloride (Flumadine, trade name: Rimantadine)), and (2) neuraminidase inhibitors as agents for inhibiting the budding/release of viruses from cells, Oseltamivir (Oseltamivir, trade name: Tamiflu, Zanamivir (Zanamivir, trade name: Relenza), peramivir (permivir), and laninamivir octanoate (laninamivir octanoate).

Despite the excellent efficacy of these existing drugs, there are still a number of problems to be overcome in treating influenza, such as: the use of existing drugs has led to the emergence of drug-resistant influenza virus strains; these drugs have considerable toxic side effects; a pandemic of a novel influenza virus with high pathogenicity or lethality may occur. Therefore, there is still a need and urgency to develop new mechanisms of anti-influenza drugs.

Cap-dependent endonucleases are influenza virus-derived enzymes, play a key role in virus propagation, and have virus-specific enzymatic activities not possessed by the host, and thus are excellent good targets for developing novel anti-influenza drugs. The cap-dependent endonuclease activity is: a fragment containing 9 to 13 bases of a cap structure (bases of the cap structure are not included in this number) is produced using a host mRNA precursor as a substrate. This fragment functions as a primer for viral RNA polymerase and is used for the synthesis of mRNA encoding viral proteins. That is, the substance which inhibits cap-dependent endonuclease inhibits synthesis of viral protein by inhibiting synthesis of viral mRNA, thereby inhibiting viral growth.

Among the substances reported to inhibit cap-dependent endonuclease are: favipiravir (approved in 2014, Furuta Y, et al, anti Res.2013; 100(2): 446-54; Vanderland E, et al, anti Agents Chemother.2016; 60(11): 6679-. In addition, compounds having a structure similar to that of the present compounds are also reported in patents and literatures as HIV antiviral and influenza antiviral (Raheem, I T, et al, j.med.chem.,2015,58(20), pp 8154-. Recently, the endonuclease inhibitor Baloxavir Marboxil (Xofluza) was approved in Japan (Hayden FG, et al, "Baloxavir Marboxil for unomplified Influenza in Adults and Adoleracens." NEngl J Med.2018; 379(10): 913-. However, the drug resistance of influenza virus inevitably appears, and small molecules with different chemical structures aiming at the same target point can have different drug resistance, so that the development of new influenza virus nuclease endonuclease inhibitors is still necessary.

Disclosure of Invention

The invention provides a compound of an azacyclic diketone compound and a prodrug thereof.

The invention provides a nitrogen heterocyclic diketone compound, which is characterized in that: is a compound represented by the following structure:

wherein the content of the first and second substances,

R¹selected from the group consisting of hydrogen, halogen, cyano, optionally substituted alkyl, amino, optionally substituted alkoxy;

R²selected from hydrogen, -C (═ O) R^A,-C(＝O)OR^A,-CH₂OC(＝O)R^A,-CH₂OC(＝O)OR^A,-CH(Me)OC(＝O)R^A,-CH(Me)OC(＝O)OR^A； R^ASelected from alkyl, substituted alkyl;

R^Aselected from alkyl, substituted alkyl;

x is selected from NR⁴,CR⁵R⁶；

R⁴Selected from the group consisting of hydrogen, alkyl, substituted alkyl, optionally substituted heterocyclyl, optionally substituted cycloalkyl;

R⁵and R⁶The same or different, independently selected from hydrogen, alkyl, substituted alkyl;

or R⁵And R⁶Together form an optionally substituted cycloalkyl group, or an optionally substituted heterocyclyl group;

R³selected from hydrogen, alkyl, substituted alkyl;

or R³And R⁴Together form an optionally substituted heterocyclyl;

or R³And R⁵Together form an optionally substituted cycloalkyl group, or an optionally substituted heterocyclyl group;

a is selected from the following tricyclic heterocyclic group, or tetracyclic heterocyclic group

Y is selected from S, S (O), O, N, CR⁸R⁹；

Ring B is selected from optionally substituted monocyclic or bicyclic heteroaryl;

ring C is selected from optionally substituted aryl, optionally substituted heteroaryl;

ring D is selected from optionally substituted aryl, optionally substituted heteroaryl;

Q¹，Q²，Q³，Q⁴and Q⁵Forming an optionally substituted five-membered aromatic heterocyclic group E;

Q¹，Q²，Q³，Q⁴and Q⁵The same or different, are independently selected from C, N, O, S;

r7 is hydrogen, halogen, cyano, alkyl, alkoxy, nitro, amino, amido;

R⁸and R⁹Independently is optionally selected from hydrogen, alkyl, substituted alkyl.

n is the number of substituents of R7 on the tricyclic, or tetracyclic, heterocyclic group. Most preferably 0-6.

Furthermore, the invention provides a nitrogen heterocyclic diketone compound, which is characterized in that: said Q¹，Q²，Q³，Q⁴And Q⁵Two or more of them are nitrogen atoms. A similar structure can be formed as follows.

In the above structure, the hydrogen bonded to the carbon atom or the N atom on the five-membered ring may be substituted by a group such as an alkyl group, an alkoxy group, or an acyl group.

Furthermore, the invention provides a nitrogen heterocyclic diketone compound, which is characterized in that: is a compound represented by the following structure:

wherein R is¹，R²，R³Ring A is shown as general formula I;

m is 0, 1,2,3, 4; i.e. a ternary, quaternary, quinary, hexahydric, heptatomic ring.

R⁴Is hydrogen optionally substituted on the polycyclic ring, alkyl, substituted alkyl, heterocyclic group optionally substituted, and cycloalkyl optionally substituted.

The saturated/unsaturated cyclic structure refers to a ring having one or more unsaturated double bonds. For example, in the form shown in the following structure or a similar structure thereto:

wherein, X, Y, R²，R³R7, n, ring C are shown as general formula I;

R₇₁and R₇₂Forming a five-membered aromatic heterocycle; or R₇₃And R₇₂Forming a five-membered heteroaromatic ring.

or

Wherein A, B, C, D, E, F is selected from nitrogen, sulfur and oxygen;

r' is hydrogen, alkyl and halogen substituted by any one or more than one of five-membered aromatic heterocyclic rings.

The five-membered aromatic ring may have a structure shown as an example below and the like:

or

Furthermore, the invention provides a nitrogen heterocyclic diketone compound which is characterized by being a compound shown in the following structure:

wherein, X, Y, R²，R³R7, n, ring C are shown as general formula I;

the ring B is a ternary, quaternary, quinary, hexahydric or heptatomic aromatic ring.

Furthermore, the invention provides a nitrogen heterocyclic diketone compound, which is characterized in that: one carbon atom on the B ring is substituted by carbonyl and oxime group.

I.e., a structure shown in the form of:

furthermore, the invention provides a nitrogen heterocyclic diketone compound, which is characterized in that: the optionally substituted alkyl refers to that hydrogen atoms on an alkyl carbon chain are substituted by hydrogen, halogen, alkyl, nitro, amino, alkenyl, alkynyl, ester group, amido, optionally substituted cycloalkyl/heterocycloalkyl and optionally substituted aryl, or carbon atoms on an alkyl carbon chain are substituted by oxygen, sulfur, nitrogen and carbonyl;

the optionally substituted amino refers to the replacement of hydrogen on the amino group by hydrogen, alkyl, optionally substituted cycloalkyl/heterocycloalkyl and optionally substituted aryl;

the optionally substituted alkoxy refers to that hydrogen atoms on an alkoxy carbon chain are substituted by hydrogen, halogen, alkyl, nitro, amino, alkenyl, alkynyl, ester group, amido, optionally substituted cycloalkyl/heterocycloalkyl and optionally substituted aryl;

the optionally substituted cycloalkyl refers to the substituent of hydrogen atom, halogen, alkyl, nitro, amino, alkenyl, alkynyl, ester group, amido, optionally substituted cycloalkyl/heterocycloalkyl and optionally substituted aryl on the alkyl ring;

the optionally substituted heterocyclic group means that hydrogen atoms on the heterocyclic ring are substituted by hydrogen, halogen, alkyl, nitro, amino, alkenyl, alkynyl, ester group, amido, optionally substituted cycloalkyl/heterocycloalkyl and optionally substituted aryl;

the optionally substituted aryl refers to that hydrogen atoms on an aryl ring are substituted by hydrogen, halogen, alkyl, nitro, amino, alkenyl, alkynyl, ester group and amide group, or carbon atoms on the aryl ring are substituted by carbonyl;

the optionally substituted heteroaryl refers to the replacement of hydrogen atoms on a heteroaromatic ring by hydrogen, halogen, alkyl, nitro, amino, alkenyl, alkynyl, ester group and amide group, or the replacement of carbon atoms on the heteroaromatic ring by carbonyl.

Furthermore, the invention provides a nitrogen heterocyclic diketone compound, which is characterized in that: one or more hydrogen atoms on the azacyclic dione compound are replaced by an isotope of hydrogen;

and/or

One or more carbon atoms on the azacyclic dione compound are replaced by an isotope of carbon;

furthermore, the invention provides a nitrogen heterocyclic diketone compound, which is characterized in that: application to the inhibition of the activity of cap-dependent endonucleases.

Detailed Description

Some embodiments are disclosed in further detail in the following examples, which are not intended to limit the scope of the claims in any way.

Synthesis of intermediates

Intermediate A5- (benzyloxy) -3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione

The specific reaction equation is as follows:

step A3- (3- (benzyloxy) -4-oxo-4H-pyran-2-yl) -3-hydroxy-2, 2-dimethylpropionaldehyde (Compound A.1)

3- (benzyloxy) -4-oxo-4H-pyran-2-carbaldehyde (10g,43mmol) was dissolved in 100mL of tetrahydrofuran, and pyrrolidine (0.35mL,17.5mmol), glacial acetic acid (3.73mL,97mmol) and isobutyraldehyde (4.3g,60mmol) were added at room temperature and stirred for 2 hours. The reaction solution was directly spin-dried and purified by column chromatography to give product A.1(9g, yield: 68.5%). LCMS ESI (+) M/z 303.1(M +1).

Step B5- (benzyloxy) -4-hydroxy-3, 3-dimethyl-3H-pyrido [1,2-B ] pyridazin-6 (4H) -one (Compound A.2)

Compound A.1(13g,43mmol) was dissolved in 200mL of N, N-dimethylformamide, hydrazine hydrochloride (30g,430mmol) was added, and the mixture was stirred at 80 ℃ for 2 hours. The reaction solution was poured into 1L of water, extracted three times with 500mL of ethyl acetate, the organic phases were combined, washed with saturated brine, and the organic phase was dried over anhydrous sodium sulfate, filtered, spun-dried, and purified by column chromatography to give product A.2(5g, yield: 38.8%). LCMS ESI (+) M/z 299.1(M +1).

Step C tert-butyl 5- (benzyloxy) -3, 3-dimethyl-6-oxo-4, 6-dihydro-3H-pyrido [1,2-b ] pyridazin-4-ylcarbonate (Compound A.3)

Compound A.2(12.5g,41mmol), triethylamine (6.35g,62.9mmol) and N, N-dimethyl-4-aminopyridine (512mg, 4.1mmol) were dissolved in 200mL of tetrahydrofuran, and a solution of di-tert-butyl dicarbonate (13.8g,63mmol) in 50mL of tetrahydrofuran was slowly added dropwise at zero degrees C, followed by stirring at room temperature for 1 hour. 500mL of water was added to the reaction mixture, and the mixture was extracted three times with 250mL of ethyl acetate, and the organic phases were combined, washed with saturated brine, and dried over anhydrous sodium sulfate. Spin-dry and purify to give product A.3(12.5g, yield: 74.9%, oil). LCMS ESI (+) M/z 399.1(M +1).

Step D tert-butyl 5- (benzyloxy) -3, 3-dimethyl-6-oxo-2, 3,4, 6-tetrahydro-1H-pyrido [1,2-b ] pyridazin-4-ylcarbonate (Compound A.4)

Compound A.3(12.5g,31.3mmol) was dissolved in 200mL of tetrahydrofuran solution, and sodium borohydride (2.2g,58.1mmol) was added in portions while cooling on ice, and stirred for 30 minutes. Quenched with saturated aqueous ammonium chloride solution, extracted three times with 250mL of ethyl acetate, the organic phases were combined, washed with saturated brine and dried over anhydrous sodium sulfate. Filtration, spin-drying and purification by column chromatography gave product A.4(12g, yield: 95.5%). LCMS ESI (+) M/z 401.1(M +1).

Step E (benzyloxy) -4- ((tert-butoxycarbonyl) oxy) -3, 3-dimethyl-6-oxo-2, 3,4, 6-tetrahydro-1H-pyrido (9H-fluoren-9-yl) o [1,2-b ] pyridazine-1-carboxylic acid tert-butyl ester (Compound A.5)

Compound A.4(5g,12.5mmol) was dissolved in 50mL of tetrahydrofuran solution, and fluorenylmethoxycarbonylcarbonyl chloride (3.9g, 15.1mmol) and a saturated aqueous solution of sodium hydrogencarbonate (10mL) were added under ice-bath, and the mixture was stirred at room temperature for 0.5 hour. The reaction was quenched with 150mL of water, extracted three times with 100mL of ethyl acetate, the organic phases were combined, washed three times with saturated brine, and the organic phase was dried over anhydrous sodium sulfate, filtered, spun-dried, and purified to give product A.5(7.9g, yield: 67%). LCMS ESI (+) M/z 622.2(M +1).

Step F (9H-fluoren-9-yl) methyl 5- (benzyloxy) -4-hydroxy-3, 3-dimethyl-6-oxo-2, 3,4, 6-tetrahydro-1H-pyrido [1,2-b ] pyridazine-1-carboxylic acid tert-butyl ester (Compound A.6)

Compound A.5(7.9g,12.7mmol) was dissolved in 40mL of dichloromethane and trifluoroacetic acid (20mL) was added dropwise under ice-bath, slowly warmed to room temperature and stirred for 30 min. The reaction solution was directly spin-dried to obtain product A.6(6g, yield: 98%). LCMS ESI (+) M/z 523.2(M +1).

Step G (9H-fluoren-9-yl) methyl 5- (benzyloxy) -3, 3-dimethyl-4, 6-dioxo-2, 3,4, 6-tetrahydro-1H-pyrido [1,2-b ] pyridine-1-carboxylic acid (compound A.7)

Compound A.6(6g,11.5mmol) was dissolved in 200mL of dichloromethane, cooled to 0 deg.C, slowly added 9.8g of dess-martin oxidant, warmed to room temperature and stirred for 1 hour. The reaction was quenched by adding sodium bicarbonate and aqueous sodium thiosulfate, extracted three times with 200mL of dichloromethane, the organic phases were combined, washed three times with saturated brine, and the organic phase was dried over anhydrous sodium sulfate. Filtration, spin-drying and purification gave product A.7(5.5g, yield: 98%). LCMS ESI (+) M/z:521.2(M +1).

Step H5- (benzyloxy) -3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione (intermediate A)

Compound A.7(5.5g,10.6mmol) was dissolved in 55mL of dichloromethane solution, 5.5mL of piperidine was added, the reaction was quenched with water while stirring at room temperature for 1 hour, extracted three times with 200mL of dichloromethane, the organic phases were combined, washed three times with saturated brine, and the organic phase was dried over anhydrous sodium sulfate. Filtration, spin-drying and purification by column chromatography gave intermediate A (2.8g, yield: 94%). LCMS ESI (+) M/z 299.1(M +1).¹H NMR(400MHz,DMSO-d6)7.74(d,J＝7.6Hz,1H),7.49–7.41(m,2H),7.35(dd,J＝8.5, 3.6Hz,3H),7.01(t,J＝7.1Hz,1H),6.23(d,J＝7.6Hz,1H),5.16(s,2H),3.20(d,J＝7.1 Hz,2H),1.11(s,6H).

The method for synthesizing intermediate A was used to synthesize intermediate B and intermediate C

Intermediate B5 '- (benzyloxy) -1', 2 '-dihydrospiro [ cyclobutane-1, 3' -pyrido [1,2-B ] pyridazine ] -4', 6' -dione

LCMS ESI(+)m/z:311.1(M+1).

Intermediate C5 '- (benzyloxy) -1', 2 '-dihydrospiro [ cyclopentane-1, 3' -pyrido [1,2-b ] pyridazine ] -4', 6' -dione

LCMS ESI(+)m/z:325.1(M+1).

Intermediate D (R) -7- (benzyloxyyl) -3,4,12,12 a-tetrahydro-1H- [1,4] pyrano [4,3-c ] pyrido [2,1-f ] [1,2,4] triazine-6, 8(1H, 3H) -dione

Intermediate D was synthesized according to the method described in JP5971830,2016, B1.LCMS ESI (+) M/z:299.1(M +1).

Intermediate E [1,2,4] triazine-4, 6-dione of (R) -5- (benzyloxy) -3- (1,1, 1-trifluoropropan-2-yl) -2, 3-dihydro-1H-pyrido [2,1-f ]

Intermediate E was synthesized according to the method described in U.S. Pat. No. 5,983, 197219,2013, A1.LCMS ESI (+) M/z:368.1(M +1).

Intermediate F [1,2,4] triazine-4, 6-dione of 5- (benzyloxy) -3-isopropyl-2, 3-dihydro-1H-pyrido [2,1-F ]

Secondly, synthesis of target compound

Example 1(2103)

1- (6, 11-dihydro-1, 2,11a triazabenzo [ cd, g ] -azulen-6-yl-5-hydroxy-3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,22-b ] pyridazine-4, 6-dione

The specific reaction equation is as follows:

step A:2- (benzylamino) -3-nitrobenzoic acid methyl ester (Compound 1.1)

1-fluoro-2-nitro-benzoic acid methyl ester (500mg, 2.5mmol) was weighed out and dissolved in 8mL of N, N-dimethylformamide, and diisopropylethylamine (650mg, 5.1mmol) and benzylamine (323mg, 3.0mmol) were added and reacted at 80 ℃ for 12 hours under nitrogen protection. To the reaction mixture was added ethyl acetate (30mL), which was washed 2 times with saturated brine (20mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (petroleum ether: ethyl acetate: 5:1) to give product 1.1(700mg, yield: 97%, yellow solid). LCMS ESI (+) M/z:287.1(M +1).

And B: 3-amino-2- (benzylamino) benzoic acid methyl ester (Compound 1.2)

Compound 1.1(700mg, 2.45mmol) is dissolved in 15mL of acetic acid, iron powder (1.37g, 24.5mmol) is added in portions under nitrogen, stirring is carried out at 25 ℃ for 0.5 hour, and then the temperature is raised to 75 ℃ and stirring is carried out for 1 hour. The reaction mixture was concentrated under reduced pressure, ethyl acetate (100mL) and water (40mL) were added to the residue, the mixture was filtered through Celite, and the filtrate was adjusted to pH 8 with sodium hydrogencarbonate. The organic phase was washed 2 times with saturated brine (20mL) and dried over anhydrous sodium sulfate. Filtration and concentration under reduced pressure, and column chromatography (petroleum ether: ethyl acetate: 5:1) of the residue afforded product 1.2(570mg, yellow oil, yield: 91%). LCMS ESI (+) M/z 257.1(M +1).

And C: 1-benzyl-1H-benzo [ d ] [1,2,3] triazole-7-carboxylic acid methyl ester (Compound 1.3)

Compound 1.2(570mg, 2.22mmol) was dissolved in 18mL of acetic acid, sodium nitrite (169mg, 2.44mmol) was added at room temperature, and the reaction mixture was stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure, and ethyl acetate (100mL) was added to the residue to adjust the pH to 8 with sodium bicarbonate solution. The organic phase was washed 2 times with saturated brine (20mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (petroleum ether: ethyl acetate ═ 3:1) to give product 1.3(580mg, white solid, yield: 97%). LCMS ESI (+) M/z 268.1(M +1).

Step D: 1-benzyl-1H-benzo [ d ] [1,2,3] triazole-7-carboxylic acid (Compound 1.4)

Compound 1.3(580mg, 2.2mmol) was dissolved in 10mL of tetrahydrofuran, and 3mL of an aqueous solution containing lithium hydroxide monohydrate (273mg, 6.5mmol) was added. The reaction solution was stirred at room temperature for 3 hours. The reaction mixture was concentrated under reduced pressure to remove tetrahydrofuran, 15mL of water was added, a saturated citric acid solution was added to adjust the pH to 5, and extraction was performed with ethyl acetate (40mLX 2). The combined organic phases were washed with saturated brine (20mL) and dried over anhydrous sodium sulfate. Filtration and concentration under reduced pressure gave the product 1.4(500mg, yield: 90%, white solid). LCMS ESI (+) M/z 254.1(M +1).

Step E:1, 2,11a triazabenzo [ cd, g ] azulene-6- (11H) -one (Compound 1.5)

Compound 1.4(500mg, 1.97mmol) was added to 5mL of polyphosphoric acid, heated to 120 ℃ under nitrogen, and stirred for 4 hours. To the reaction mixture were added water (30mL) and ethyl acetate (30 mL). The organic phase was adjusted to pH 9 with saturated sodium bicarbonate solution, washed with saturated brine (20mL) and dried over anhydrous sodium sulfate. Filtration and concentration under reduced pressure gave crude product 1.5(400mg, yellow solid). LCMS ESI (+) M/z:236.1(M +1)

Step F: 6, 11-dihydro-1, 2,11a triazabenzo [ cd, g ] azulen-6-ol (Compound 1.6)

Compound 1.5(400mg, 1.7mmol) was dissolved in 8mL tetrahydrofuran and sodium borohydride (193mg, 5.1mmol) was added at 0 deg.C. The reaction mixture was warmed to room temperature and stirred for 0.5 hour. The reaction mixture was quenched by addition of saturated ammonium chloride solution (5mL), extracted with ethyl acetate (40mL), and the organic phase was washed with saturated brine (15mL) and dried over anhydrous sodium sulfate. Filtration and concentration under reduced pressure, and the residue was purified by column chromatography (petroleum ether: ethyl acetate 1:1) to give 1.6(250mg, yield: 62%) as a yellow solid. LCMS ESI (+) M/z:238.1(M +1).

Step G: 5- (benzyloxy) -1- (6, 11-dihydro-1, 2,11a triazabenzo [ cd, g ] azulen-6-yl) -3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione (Compound 1.7)

Compound 1.6(130mg, 0.55mmol) and 5- (benzyloxy) -3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione (136mg, 0.45mmol) were dissolved in 2mL of 1-propylphosphoric anhydride/N, N-dimethylformamide (50%). The reaction solution was stirred at 110 ℃ under nitrogen for 1 hour. Water (10mL) was added to the reaction mixture, which was extracted 2 times with ethyl acetate (30mL), and the combined organic phases were washed with saturated brine (15mL) and dried over anhydrous sodium sulfate. Filtration and concentration under reduced pressure, and column chromatography (petroleum ether: ethyl acetate: 10:1) of the residue yielded 1.7(130mg, yield: 55%) as a white solid product. LCMS ESI (+) M/z:518.2(M +1).

Step H: 1- (6, 11-dihydro-1, 2,11a triazabenzo [ cd, g ] azulen-6-yl) -5-hydroxy-3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,22-b ] pyridazine-4, 6-dione (example 1)

Compound 1.7(130mg, 0.25mmol) was dissolved in 4mLN, N-dimethylformamide and lithium chloride (532mg, 12.6mmol) was added. The reaction solution was stirred at 100 ℃ under nitrogen for 5 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by reverse phase preparative purification to give the product of example 1 (47mg, pale yellow solid, yield: 44%). LCMS ESI (+) M/z 428.1(M +1).¹H NMR(400MHz,DMSO-d₆)7.95(d, J＝8.4Hz,1H),7.75(dd,J＝7.2,1.6Hz,1H),7.62(d,J＝7.2Hz,1H),7.48-7.40(m,2H), 7.15(dd,J＝8.4,7.2Hz,1H),7.05(d,J＝6.8Hz,1H),6.77(d,J＝7.6Hz,1H),6.69(d,J＝ 14.4Hz,1H),6.12(d,J＝14.4Hz,1H),5.49(s,1H),5.42(d,J＝7.6Hz,1H),3.58(d,J＝14.8 Hz,1H),3.28(d,J＝14.8Hz,1H),1.47(s,3H),1.10(s,3H).

Referring to example 1, the two-step synthesis, replacing intermediate A with intermediates B and C, gave examples 2 and 3

Example 2(2180)

1' - (6, 11-dihydro-1, 2,11a triazabenzo [ cd, g ] azulen-6-yl) -5' -hydroxy-1 ', 2' -dihydrospiro [ cyclobutane-1, 3' -pyrido [1,2-b ] pyridazine ] -4', 6' -dione

LCMS ESI(+)m/z:440.2(M+1).

Example 3(2181)

1' - (6, 11-dihydro-1, 2,11a triazabenzo [ cd, g ] azulen-6-yl) -5' -hydroxy-1 ', 2' -dihydrospiro [ cyclopentane-1, 3' -dihydropyrido [1,2-b ] pyridazine ] -4', 6' -dione

LCMS ESI(+)m/z:454.2(M+1).

Example 4(2114)

5-hydroxy-3, 3-dimethyl-1- (2-methyl-2, 6-dihydrobenzo [6,7] thiaheptacyclo [2,3,4-c, d ] indazol-6-yl) -2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione

The specific reaction equation is as follows:

step A: 1H-indazole-4-carboxylic acid methyl ester (Compound 4.1)

Methyl 2-methyl-3-amino-benzoate (5g, 30.3mmol) was dissolved in 40mL of acetic acid, and sodium nitrite (2.3g, 33.3mmol) was added. The reaction was carried out at 25 ℃ under nitrogen for 1 hour. The reaction mixture was concentrated under reduced pressure to remove acetic acid, and ethyl acetate (80mL) was added to the residue, which was adjusted to pH 8 with a saturated sodium bicarbonate solution, washed 2 times with saturated brine (20mL), and dried over anhydrous sodium sulfate. Filtration and concentration under reduced pressure, the residue was purified by column chromatography (petroleum ether: ethyl acetate ═ 3:1) to give 4.1(2.4g, yellow solid, yield: 45%. LCMS ESI (+) M/z:177.1(M +1).

And B: 3-bromo-1H-indazole-4-carboxylic acid methyl ester (Compound 4.2)

Compound 4.1(2.4g, 13.6mmol) was dissolved in 40mL of N, N-dimethylformamide, potassium hydroxide (3.1g, 54.5mmol) and N-bromosuccinimide (4.9g, 27.2mmol) were added, and the mixture was stirred at 25 ℃ under nitrogen for 1 hour. Water (50mL) was added to the reaction mixture, and the mixture was extracted 3 times with ethyl acetate (60 mL). The combined organic phases were washed 2 times with saturated sodium bicarbonate solution (30mL) and 2 times with saturated brine solution (30mL) and dried over anhydrous sodium sulfate. Filtration and concentration under reduced pressure, and column chromatography (petroleum ether: ethyl acetate: 3:1) of the residue afforded 4.2(2.3g, yellow oil, yield: 66%). LCMS ESI (+) M/z 255.0,257.0(M +1).

And C: 3-bromo-1-methyl-1H-indazole-4-carboxylic acid methyl ester (Compound 4.3)

Compound 4.2(2g, 7.8mmol) was dissolved in 25mL of N, N-dimethylformamide, sodium hydride (627mg, 15.7mmol) was added at 0 degrees, the reaction solution was stirred at 0 degrees for 0.5 hour, and methyl iodide (2.23g, 15.7mmol) was then slowly added dropwise. The reaction solution was stirred at room temperature for 2 hours. Water (20mL) was added to the reaction solution, and the mixture was extracted 2 times with ethyl acetate (60 mL). The combined organic phases were washed 3 times with saturated brine (20mL) and dried over anhydrous sodium sulfate. Filtration and concentration under reduced pressure, and column chromatography (petroleum ether: ethyl acetate: 3:1) of the residue yielded 4.3(2.2g, yellow oil, yield: 95%). LCMS ESI (+) M/z 269.0,271.0(M +1).

Step D: 1-methyl-3- (phenylthio) -1H-indazole-4-carboxylic acid (Compound 4.4)

Compound 4.3(2.2g, 8.2mmol) was dissolved in 30mL of isopropanol and 10mL of ethylene glycol, and potassium carbonate (2.3g, 16.4mmol), thiophenol (1.8g, 16.4mmol) and cuprous iodide (778mg, 4.1mmol) were added. The reaction solution was stirred at 100 ℃ under nitrogen for 12 hours. To the reaction mixture were added water (50mL) and ethyl acetate (50 mL). The aqueous phase was washed 2 times with ethyl acetate (40mL) and the pH was adjusted to 4 with saturated citric acid. The mixture was extracted 2 times with ethyl acetate (60mL), and the combined organic phases were washed with saturated brine (30mL) and dried over anhydrous sodium sulfate. Filtration and concentration under reduced pressure gave 4.4(1.8g, yellow solid, yield: 77%). LCMS ESI (-) M/z 283.1(M-1).

Step E: 2-methylbenzo [6,7] thiaheptacyclo [2,3,4-cd ] indazol-6- (2H) -one (Compound 4.5)

Compound 4.4(1.8g, 6.33mmol) was added to 8mL of polyphosphoric acid and stirred at 120 ℃ under nitrogen for 2 hours. After cooling, 30mL of water and 30mL of ethyl acetate were added to the reaction mixture. The organic phase was adjusted to pH 9 with saturated sodium bicarbonate solution, washed with saturated brine (20mL) and dried over anhydrous sodium sulfate. Filtration and concentration under reduced pressure gave crude product 4.5(960mg, yellow solid). LCMS ESI (+) M/z:267.1(M +1)

Step F: 2-methyl-2, 6-dihydrobenzo [6,7] thiaheptacyclo [2,3,4-cd ] indazol-6-ol (Compound 4.6)

Compound 4.5(960mg, 3.6mmol) was dissolved in 10mL tetrahydrofuran and sodium borohydride (409mg, 10.8mmol) was added at 0 deg.C. The reaction solution was stirred at 0 ℃ for 1 hour. The reaction mixture was quenched by adding saturated ammonium chloride solution (20mL), extracted 2 times with ethyl acetate (40mL), and the combined organic phases were washed with saturated brine (20mL) and dried over anhydrous sodium sulfate. Filtration and concentration under reduced pressure, and column chromatography of the residue (petroleum ether: ethyl acetate: 1) gave 4.6(320mg, yellow solid, yield: 33%). LCMS ESI (+) M/z 269.1(M +1).

Step G: 5- (benzyloxy) -3, 3-dimethyl-1- (2-methyl-2, 6-dihydrobenzo [6,7] thiaheptacyclo [2,3,4-cd ] indazol-6-yl) -2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione (Compound 4.7)

Compound 4.6(50mg, 0.17mmol) and 5- (benzyloxy) -3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazin-4, 6-dione (54mg, 0.21mmol) were dissolved in 3mL of 1-propylphosphoric anhydride/N, N-dimethylformamide (50%). The reaction solution was stirred at 100 ℃ under nitrogen for 1 hour. Water (10mL) was added to the reaction mixture, which was extracted 2 times with ethyl acetate (30mL), and the combined organic phases were washed with saturated brine (15mL) and dried over anhydrous sodium sulfate. Filtration and concentration under reduced pressure, and column chromatography (petroleum ether: ethyl acetate: 10:1) of the residue afforded 4.7(47mg, white solid, yield: 51%). LCMS ESI (+) M/z 549.2(M +1).

Step H: 5-hydroxy-3, 3-dimethyl-1- (2-methyl-2, 6-dihydrobenzo [6,7] thiaheptacyclo [2,3,4-cd indazol-6-yl) -2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione (example 4)

Compound 4.7(47mg, 0.09mmol) was dissolved in 3mLN, N-dimethylformamide and lithium chloride (115mg, 2.6mmol) was added. The reaction solution was stirred at 100 ℃ under nitrogen for 1 hour. The reaction solution was concentrated under reduced pressure, and the residue was purified by reverse phase preparative purification to give the product, example 4(7mg, yellow solid, yield: 18%). LCMS ESI (+) M/z 459.1(M +1).

Example 5 (2131)

5-hydroxy-3, 3-dimethyl-1- (1-methyl-6, 12-dihydro-1H-benzo [6',7' ] thiaheptacyclo [3',4':3,4] benzo [1,2-d ] imidazol-6-yl) -2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione

The specific reaction equation is as follows:

step A: 4-bromo-3-methylbenzene-1, 2-diamine (Compound 5.1)

2-Nitro-3-methyl-4-bromo-aniline (2.6g, 11.3mmol) was weighed, dissolved in 30mL ethanol and 15mL water, ammonium chloride (6g, 112.5mmol) was added, then iron powder (2.5g, 45.0mmol) was added in portions and reacted at 90 ℃ under nitrogen for 1 hour. The reaction solution was cooled to room temperature and filtered through celite. Ethyl acetate (100mL) was added to the filtrate, which was washed with saturated brine (30mL) 2 times and dried over anhydrous sodium sulfate. Filtration and concentration under reduced pressure, and the residue was purified by column chromatography (petroleum ether: ethyl acetate: 3:1) to give 5.1(2g, yellow oil, yield: 88%). LCMS ESI (+) M/z 201.0,203.0(M +1).

And B: 6-bromo-7-methyl-1H-benzo [ d ] imidazole (Compound 5.2)

Compound 5.1(2g, 10.0mmol) was dissolved in 20mL of acetic acid and stirred at 0 ℃ under nitrogen for 12 hours. The reaction mixture was concentrated under reduced pressure, and to the residue was added saturated sodium bicarbonate to adjust the pH to 8, extracted 2 times with ethyl acetate (60mL), the organic phases were combined, washed 2 times with saturated brine (20mL), and dried over anhydrous sodium sulfate. Filtration and concentration under reduced pressure gave crude product 5.2(2.1g, yellow solid). LCMS ESI (+) M/z 210.0,212.0(M +1).

And C: 6-bromo-1, 7-dimethyl-1H-benzo [ d ] imidazole (Compound 5.3)

Compound 5.2(2.1g, 9.9mmol) was dissolved in 25mL N, dimethylformamide and sodium hydride (796mg, 19.9mmol) was added at 0 deg.C. The reaction solution was stirred at 0 ℃ under nitrogen for 0.5 hour. Methyl iodide (2.82g, 19.9mmol) was then added slowly dropwise. The reaction solution was stirred at room temperature for 2 hours. Water (30mL) was added to the reaction mixture, which was extracted 2 times with ethyl acetate (60mL), and the combined organic phases were washed 3 times with saturated brine (20mL) and dried over anhydrous sodium sulfate. Filtration and concentration under reduced pressure, and column chromatography (petroleum ether: ethyl acetate: 10:1) of the residue yielded 5.3(1.6g, yellow oil, yield: 71%). LCMS ESI (+) M/z 225.0,227.0(M +1).

Step D: methyl-1, 7-dimethyl-1H-benzo [ d ] imidazole-6-carboxylic acid methyl ester (Compound 5.4)

Compound 5.3(1.4g, 6.22mmol) was dissolved in 6mL of N, N-dimethylformamide, 20mL of methanol and 6mL of triethylamine, and 1, 1-bis-diphenylphosphinoferrocene palladium dichloride (700mg, 0.95mmol) and dicobalt octacarbonyl (1.4g, 4.09mmol) were added. The reaction solution was stirred at 80 ℃ under nitrogen for 12 hours. The reaction mixture was filtered through celite, the filtrate was concentrated under reduced pressure, and the residue was dissolved in ethyl acetate (100mL), washed 3 times with saturated brine (30mL), and dried over anhydrous sodium sulfate. Filtration and concentration under reduced pressure, and column chromatography of the residue (petroleum ether: ethyl acetate: 10:1) gave 5.4(1g, brown oil, yield: 78%). LCMS ESI (+) M/z:205.1(M +1).

Step E: 7- (bromomethyl) -1-methyl-1H-benzo [ d ] imidazole-6-carboxylic acid methyl ester (Compound 5.5)

Compound 5.4(1g, 4.9mmol) was added to 25mL of carbon tetrachloride, followed by N-bromosuccinimide (915mg, 5.1mmol) and benzoyl peroxide (316mg,0.98 mmol). The reaction solution was stirred at 78 ℃ under nitrogen for 12 hours. Methylene chloride (50mL) and water (30mL) were added to the reaction mixture. The organic phase was washed 2 times with saturated brine (20mL) and dried over anhydrous sodium sulfate. Filtration and reduced pressure concentration of the residue were performed, and the residue was purified by column chromatography (petroleum ether: ethyl acetate: 10:1) to give 5.5(380mg, yellow solid, yield: 27%). LCMS ESI (+) M/z:283.0,285.0(M +1)

Step F: 1-methyl-7- ((phenylthio) methyl) -1H-benzo [ d ] imidazole-6-carboxylic acid methyl ester (Compound 5.6)

Compound 5.5(380mg, 1.3mmol) was dissolved in 8mL of N, N-dimethylformamide, and thiophenol (178mg, 1.6mmol) and potassium carbonate (370mg, 2.7mmol) were added. The reaction solution was stirred at room temperature for 2 hours. Water (20mL) was added to the reaction mixture, which was extracted 2 times with ethyl acetate (40mL), and the combined organic phases were washed 2 times with saturated brine (20mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the crude product 5.6(420mg, yellow oil). LCMS ESI (+) M/z:313.1(M +1).

Step G: 1-methyl-7- ((phenylthio) methyl) -1H-benzo [ d ] imidazole-6-carboxylic acid (Compound 5.7)

Compound 5.6(420mg, 1.3mmol) was dissolved in 6mL of tetrahydrofuran, and an aqueous solution (3mL) of lithium hydroxide (226mg, 2.4mmol) was added. The reaction mixture was stirred at 50 ℃ for 12 hours. The reaction mixture was concentrated under reduced pressure to remove tetrahydrofuran, 30mL of water was added, and the mixture was washed with ethyl acetate (15mL) 2 times. The aqueous phase was adjusted to pH 5 with saturated citric acid and extracted 2 times with ethyl acetate (40 mL). The combined organic phases were washed with saturated brine (20mL) and dried over anhydrous sodium sulfate. Filtration and concentration under reduced pressure gave product 5.7(150mg, white solid, yield: 37%). LCMS ESI (-) M/z 297.1(M-1).

Step H: 1-methyl-1H-benzo [6',7' ] thiaheptacyclo [3',4':3,4] benzo [1,2-d ] imidazol-6- (12H) -one (compound 5.8)

Compound 5.7(150mg, 0.5mmol) was added to 4mL of polyphosphoric acid and stirred at 120 ℃ under nitrogen for 1 hour. To the reaction mixture were added water (30mL) and ethyl acetate (30 mL). The organic phase was adjusted to pH 9 with saturated sodium bicarbonate solution, washed with saturated brine (20mL) and dried over anhydrous sodium sulfate. Filtration and concentration under reduced pressure gave crude product 5.8(120mg, brown solid). LCMS ESI (+) M/z 281.1(M +1).

Step I: 1-methyl-6, 12-dihydro-1H-benzo [6',7' ] thiaheptacyclo [3',4':3,4] benzo [1,2-d ] imidazol-6-ol (Compound 5.9)

Compound 5.8(120mg, 0.4mmol) was dissolved in 4mL of tetrahydrofuran and 2mL of methanol, and sodium borohydride (100mg, 2.6mmol) was added at 0 deg.C. The reaction solution was stirred at room temperature for 1 hour. To the reaction mixture was added a saturated ammonium chloride solution (10mL), and the mixture was extracted with ethyl acetate (40mL), and the organic phase was washed with a saturated brine (15mL) and dried over anhydrous sodium sulfate. Filtration and concentration under reduced pressure, and column chromatography (petroleum ether: ethyl acetate: 0:1) of the residue afforded 5.9(59mg, yellow solid, yield: 49%). LCMS ESI (+) M/z 283.1(M +1).

Step J: 5- (benzyloxy) -3, 3-dimethyl-1- (1-methyl-6, 12-dihydro-1H-benzo [6',7' ] thiaheptacyclo [3',4':3,4] benzo [1,2,2-d ] imidazol-6-yl) -2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione (Compound 5.10)

Compound 5.9(57mg, 0.20mmol) and 5- (benzyloxy) -3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione (50mg,0.17 mmol) were dissolved in 2mL of 1-propylphosphoric anhydride/N, N-dimethylformamide (50%). The reaction solution was stirred at 100 ℃ under nitrogen for 1 hour. Water (10mL) was added to the reaction solution, extracted 2 times with ethyl acetate (30mL), and the combined organic phases were washed with saturated brine (15mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane: methanol ═ 20:1) to give 5.10(100mg, yellow solid, crude). LCMSISI (+) M/z 563.2(M +1).

Step K: 5-hydroxy-3, 3-dimethyl-1- (1-methyl-6, 12-dihydro-1H-benzo [6',7' ] thiaheptacyclo [3',4':3,4] benzo [1,2-d ] imidazol-6-yl) -2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione (example 5)

Compound 5.10(100mg, 0.18mmol) was dissolved in 4mL of N, N-dimethylformamide, and lithium chloride (150mg, 7.2mmol) was added. The reaction solution was stirred at 100 ℃ under nitrogen for 2 hours. The reaction solution was concentrated under reduced pressure, and the residue was prepared using a reverse phase and purified to give the product example 5(50mg, yellow solid, yield: 59%). LCMS ESI (+) M/z 473.2(M +1).¹H NMR(400MHz,MeODL-d₄)9.21(s, 1H),7.75(d,J＝8.4Hz,1H),7.64(dd,J＝8.4,11.6Hz,2H),7.18-7.04(m,2H),6.96-6.80(m, 2H),6.07-5.90(m,2H),5.52(s,1H),4.42(d,J＝13.6Hz,1H),4.14-4.01(m,3H),3.79(d, J＝15.2Hz,1H),3.56-3.46(m,1H),1.57(s,3H),1.21(s,3H)

Example 6(2132)

1- (6, 12-Dihydrobenzo [6',7' ] thiaheptacyclo [3',4':3,4] benzo [1,2-C ] [1,2,5] thiadiazol-6-yl) -5-hydroxy-3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione

The specific reaction equation is as follows:

step A: 5-bromo-4-methylbenzo [ d ] [1,2,5] thiadiazole (Compound 6.1)

4-bromo-3-methyl-phenylenediamine (2.0g, 9.9mmol) was weighed out and dissolved in 25mL of chloroform, triethylamine (4g, 39.8mmol) was added, followed by dropwise addition of thionyl chloride (3.55g, 29.9mmol), and the reaction was carried out at 60 ℃ under nitrogen for 12 hours. The reaction mixture was concentrated under reduced pressure, and ethyl acetate (80mL) and water (40mL) were added to the residue. The organic phase was washed 2 times with saturated brine (30mL) and dried over anhydrous sodium sulfate. Filtration and concentration under reduced pressure, and column chromatography of the residue (petroleum ether: ethyl acetate: 20:1) gave product 6.1(2g, 87% yield) as a white solid.

And B: methyl 4-methylbenzo [ c ] [1,2,5] thiadiazole-5-carboxylate (Compound 6.2)

Compound 6.1(1.9g, 8.3mmol) was dissolved in 6mL of N, N-dimethylformamide, 20mL of methanol, 6mL of triethylamine, and 1, 1-bis-diphenylphosphinoferrocene palladium dichloride (1g, 1.36mmol) and dicobalt octacarbonyl (1.6g, 4.7mmol) were added. The reaction solution was stirred at 80 ℃ for 12 hours under nitrogen protection. The reaction solution was filtered through celite, the filtrate was concentrated under reduced pressure, the residue was dissolved in ethyl acetate (120mL), washed 3 times with saturated brine (30mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (petroleum ether: ethyl acetate: 30:1) to give 6.2(1.3g, yellow solid, yield: 75%).

And C: 4- (bromomethyl) benzo [ c ] [1,2,5] thiadiazole-5-carboxylic acid methyl ester (Compound 6.3)

Compound 6.2(1.3g, 6.2mmol) was added to 30mL of carbon tetrachloride, and N-bromosuccinimide (1.31g, 6.9mmol) and benzoyl peroxide (431mg, 1.2mmol) were added. The reaction solution was stirred at 78 ℃ under nitrogen for 12 hours. Dichloromethane (50mL) and water (30mL) were added to the reaction mixture. The organic phase was washed 2 times with saturated brine (20mL) and dried over anhydrous sodium sulfate. The residue was filtered and concentrated under reduced pressure and purified by column chromatography (petroleum ether: ethyl acetate: 30:1) to give 6.3(1.5g, yellow solid, yield: 84%).

Step D: ((phenylthio) methyl) benzo [ c ] [1,2,5] thiadiazole-5-carboxylic acid methyl ester (Compound 6.4)

Compound 6.3(1.5g, 5.2mmol) was dissolved in 25mL of N, N-dimethylformamide, and thiophenol (690mg, 6.3mmol) and potassium carbonate (1.44g, 10.4mmol) were added. The reaction solution was stirred at room temperature for 1 hour. Water (20mL) was added to the reaction mixture, which was extracted 2 times with ethyl acetate (40mL), and the combined organic phases were washed 2 times with saturated brine (20mL) and dried over anhydrous sodium sulfate. Filtration and concentration under reduced pressure gave 6.4(1.8g, yellow oil, crude).

Step E: 4- ((phenylthio) methyl) benzo [ c ] [1,2,5] thiadiazole-5-carboxylic acid (Compound 6.5)

Compound 6.4(1.8g, 5.7mmol) was dissolved in 20mL of tetrahydrofuran, and a solution of lithium hydroxide (716mg, 17.1mmol) in water (10mL) was added. The reaction mixture was stirred at 25 ℃ for 12 hours. The reaction mixture was concentrated under reduced pressure to remove tetrahydrofuran, 30mL of water was added, and the mixture was washed with ethyl acetate (15mL) 2 times. The aqueous phase was adjusted to pH 5 with saturated citric acid and extracted 2 times with ethyl acetate (40 mL). The combined organic phases were washed with saturated brine (20mL) and dried over anhydrous sodium sulfate. Filtration and concentration under reduced pressure gave 6.5(630mg, yellow solid, yield: 36%). LCMS ESI (-) M/z 301.0(M-1).

Step F: benzo [6',7' ] thiaheptacyclo [3',4':3,4] benzo [1,2-c ] [1,2,5] thiadiazol-6- (12H) -one (Compound 6.6)

Compound 6.5(200mg, 0.66mmol) was added to 3mL of polyphosphoric acid and stirred at 120 ℃ under nitrogen for 0.5 hour. To the reaction mixture were added water (30mL) and ethyl acetate (30 mL). The organic phase was adjusted to pH 9 with saturated sodium bicarbonate solution, washed with saturated brine (20mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the product 6.6(190mg, brown solid, crude.)

Step G: 6, 12-dihydrobenzo [6',7' ] thiaheptacyclo [3',4':3,4] benzo [1,2-C ] [1,2,5] thiadiazol-6-ol (Compound 6.7)

Compound 6.6(190mg, 0.66mmol) was dissolved in 4mL of tetrahydrofuran, and at 0 deg.C, sodium borohydride (70mg, 1.98mmol) and 1mL of methanol were added. The reaction solution was stirred at room temperature for 1 hour. To the reaction mixture was added a saturated ammonium chloride solution (10mL), and the mixture was extracted with ethyl acetate (40 ═ mL), and the organic phase was washed with a saturated brine (15mL) and dried over anhydrous sodium sulfate. Filtration and concentration under reduced pressure, and column chromatography (petroleum ether: ethyl acetate: 3:1) of the residue afforded 6.7(160mg, yellow solid, yield: 83%).

Step H:5- (benzyloxy) -1- (6, 12-dihydrobenzo [6',7' ] thiaheptacyclo [3',4':3,4] benzo [1,2-c ] [1,2,5] thiadiazol-6-yl) -3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione (Compound 6.8)

Compound 6.7(58mg, 0.20mmol) and compound 5- (benzyloxy) -3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione (50mg,0.17 mmol) were dissolved in 2mL of 1-propylphosphoric anhydride/N, N-dimethylformamide (50%). The reaction solution was stirred at 100 ℃ under nitrogen for 0.5 hour. Water (10mL) was added to the reaction mixture, which was extracted 2 times with ethyl acetate (30mL), and the combined organic phases were washed with saturated brine (15mL) and dried over anhydrous sodium sulfate. Filtration and concentration under reduced pressure, and column chromatography (petroleum ether: ethyl acetate 1:1) of the residue afforded 6.8(50mg, yellow solid, yield: 52%). LCMS ESI (+) M/z 567.1(M +1).

Step I:1- (6, 12-Dihydrobenzo [6',7' ] thiaheptacyclo [3',4':3,4] benzo [1,2-c ] [1,2,5] thiadiazol-6-yl) -5-hydroxy-3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione (example 6)

Compound 6.8(50mg, 0.09mmol) was dissolved in 3mL of N, N-dimethylformamide, and lithium chloride (200mg, 4.76mmol) was added. The reaction solution was stirred at 100 ℃ under nitrogen for 3 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by reverse phase preparative purification to give the product, example 6(11mg, yellow solid, yield: 26%). LCMS ESI (+) M/z 477.1(M +1).

Example 7(2134)

(11AS) -11- (6, 12-dihydrobenzo [6',7' ] thiaheptacyclo [3',4':3,4] benzo [1,2-c ] [1,2,5] thiadiazol-6-yl) -6-hydroxymethyl-4, 4a,11, 11A-tetrahydropyrano [4,3-e ] pyrido [1,2-b ] pyridazine-5, 7(1H, 3H) -dione

The specific reaction equation is as follows:

step A: (11AS) -6- (benzyloxy) -11- (6, 12-dihydrobenzo [6',7' ] thiaheptacyclo [3',4':3,4] benzo [1,2-c ] [1,2,5] thiadiazol-6-yl) -4,4a,11,11 a-tetrahydropyrano [4,3-e ] pyrido [1,2-b ] pyridazine-5, 7(1H, 3H) -dione (Compound 7.1)

Compound 6.7(50mg, 0.17mmol) and intermediate D (50mg, 0.16mmol) were dissolved in 2mL of 1-propylphosphoric anhydride (50% in DMF). The reaction solution was stirred at 100 ℃ under nitrogen for 0.5 hour. Water (10mL) was added to the reaction mixture, and the mixture was extracted 2 times with ethyl acetate (30 mL). The combined organic phases were washed with saturated brine (15mL) and dried over anhydrous sodium sulfate. Filtration and concentration under reduced pressure, and the residue was purified by column chromatography (petroleum ether: ethyl acetate 1:1) to give the product 7.1(30mg, yellow solid, yield: 30%). LCMS ESI (+) M/z 595.1(M +1).

And B: (11AS) -11- (6, 12-Dihydrobenzo [6',7' ] thiaheptacyclo [3',4':3,4] benzo [1,2-c ] [1,2,5] thiadiazol-6-yl) -6-hydroxymethyl-4, 4a,11, 11A-tetrahydropyrano [4,3-e ] pyrido [1,2-b ] pyridazine-5, 7(1H, 3H) -dione (Compound 7)

Compound 7.1(30mg, 0.05mmol) was dissolved in 3mL of N, N-dimethylformamide, and lithium chloride (200mg, 4.76mmol) was added. The reaction solution was stirred at 100 ℃ under nitrogen for 3 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by reverse phase preparative purification to give the product, example 7(2 mg, yellow solid, yield: 8%). LCMS ESI (+) M/z 505.0(M +1).

Example 8(2166)

1- (5, 11-Dihydrobenzo [6',7' ] thiaheptacyclo [3',4': 4,5] benzo [1,2-c ] [1,2,5] thiadiazol-11-yl) -5-hydroxy-3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione

The specific reaction equation is as follows:

step A: 5-bromo-6-methylbenzo [ d ] [1,2,5] thiadiazole (Compound 8.1)

4-bromo-5-methylbenzene-1, 2-diamine (1.0g, 5.0mmol) was weighed, dissolved in 12mL of chloroform, triethylamine (2g, 19.9mmol) was added, thionyl chloride (1.8g, 15.99mmol) was added dropwise, and the mixture was reacted at 60 ℃ under nitrogen for 12 hours. The reaction solution was concentrated under reduced pressure, and ethyl acetate (40mL) and water (20mL) were added to the residue. The organic phase was washed 2 times with saturated brine (30mL) and dried over anhydrous sodium sulfate. Filtration and concentration under reduced pressure, and the residue was purified by column chromatography (petroleum ether: ethyl acetate: 20:1) to give the product 8.1(890mg, white solid, yield: 90%).

And B: methyl 6-methylbenzo [ c ] [1,2,5] thiadiazole-5-carboxylate (Compound 8.2)

Compound 8.1(890mg, 3.89mmol) was dissolved in 3mL of N, N-dimethylformamide, 20mL of methanol and 3mL of triethylamine, followed by the addition of 1, 1-bis-diphenylphosphinoferrocene palladium dichloride (500mg, 0.68mmol) and dicobalt octacarbonyl (800mg, 2.33 mmol). The reaction solution was stirred at 80 ℃ under nitrogen for 12 hours. The reaction mixture was filtered through celite, the filtrate was concentrated under reduced pressure, and the residue was dissolved in ethyl acetate (60mL), washed 3 times with saturated brine (30mL), and dried over anhydrous sodium sulfate. Filtration and concentration under reduced pressure, and column chromatography of the residue (petroleum ether: ethyl acetate: 30:1) gave 8.2(200mg, yellow solid, yield: 25%).

And C: 6- (bromomethyl) benzo [ c ] [1,2,5] thiadiazole-5-carboxylic acid methyl ester (Compound 8.3)

Compound 8.2(200mg, 1mmol) was added to 10mL of carbon tetrachloride, and N-bromosuccinimide (190mg, 1.1mmol) and benzoyl peroxide (50mg,0.2 mmol) were added. The reaction solution was stirred at 78 ℃ under nitrogen for 12 hours. Methylene chloride (20mL) and water (10mL) were added to the reaction mixture. The organic phase was washed 2 times with saturated brine (20mL) and dried over anhydrous sodium sulfate. Filtration and concentration of the residue under reduced pressure followed by column chromatography (petroleum ether: ethyl acetate: 30:1) gave 8.3(150mg, yellow solid, yield: 51%).

Step D: 6- ((phenylthio) methyl) benzo [ c ] [1,2,5] thiadiazole-5-carboxylic acid methyl ester (Compound 8.4)

Compound 8.3(150mg, 0.51mmol) was dissolved in 5mL of N, N-dimethylformamide, and thiophenol (68mg, 0.62 mmol) and potassium carbonate (140mg, 1.02mmol) were added. The reaction solution was stirred at room temperature for 1 hour. Water (10mL) was added to the reaction mixture, which was extracted 2 times with ethyl acetate (20mL), and the combined organic phases were washed 2 times with saturated brine (20mL) and dried over anhydrous sodium sulfate. Filtration and concentration under reduced pressure gave 8.4(150mg, yellow oil, crude).

Step E: 6- ((phenylthio) methyl) benzo [ c ] [1,2,5] thiadiazole-5-carboxylic acid (Compound 8.5)

Compound 8.4(150mg, 0.5mmol) was dissolved in 10mL of tetrahydrofuran, and an aqueous solution (3mL) of lithium hydroxide (60mg, 1.5mmol) was added. The reaction mixture was stirred at 25 ℃ for 12 hours. The reaction mixture was concentrated under reduced pressure to remove tetrahydrofuran, 15mL of water was added, and the mixture was washed with ethyl acetate (10mL) 2 times. The aqueous phase was adjusted to pH 5 with saturated citric acid and extracted 2 times with ethyl acetate (20 mL). The combined organic phases were washed with saturated brine (20mL) and dried over anhydrous sodium sulfate. Filtration and concentration under reduced pressure gave 8.5(120mg, yellow solid, yield: 38%). LCMS ESI (-) M/z 301.0(M-1).

Step F: benzo [6',7' ] thiaheptacyclo [3',4': 4,5] benzo [1,2-C ] [1,2,5] thiadiazole 11(5H) -one (Compound 8.6)

Compound 8.5(120mg, 0.39mmol) was added to 3mL of polyphosphoric acid and stirred at 120 ℃ under nitrogen for 0.5 hour. To the reaction mixture were added water (30mL) and ethyl acetate (30 mL). The organic phase was adjusted to pH 9 with saturated sodium bicarbonate solution, washed with saturated brine (20mL) and dried over anhydrous sodium sulfate. Filtration and concentration under reduced pressure gave product 8.6(60mg, brown solid, crude).

Step G: 5, 11-dihydrobenzo [6',7' ] thiaheptacyclo [3',4': 4,5] benzo [1,2-c ] [1,2,5] thiadiazol-11-ol (Compound 8.7)

Compound 8.6(60mg, 0.21mmol) was dissolved in 4mL tetrahydrofuran, and at zero degrees, sodium borohydride (16mg, 0.42mmol) and 1mL methanol were added. The reaction solution was stirred at room temperature for 1 hour. To the reaction mixture was added a saturated ammonium chloride solution (10mL), and the mixture was extracted with ethyl acetate (40mL), and the organic phase was washed with saturated brine (15mL) and dried over anhydrous sodium sulfate. Filtration and concentration under reduced pressure, and column chromatography of the residue (petroleum ether: ethyl acetate: 3:1) gave 8.7(60mg, yellow solid, yield: 99%).

Step H:5- (benzyloxy) -1- (5, 11-dihydrobenzo [6',7' ] thiaheptacyclo [3',4': 4,5] benzo [1,2-c ] [1,2,5] thiadiazol-11-yl) -3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione (Compound 8.8)

Compound 8.7(50mg, 0.171mmol) and intermediate A (50mg,0.17 mmol) were dissolved in 2mL 1-propylphosphoric anhydride (50% in DMF). The reaction solution was stirred at 100 ℃ under nitrogen for 0.5 hour. Water (10mL) was added to the reaction mixture. The mixture was extracted 2 times with ethyl acetate (30mL), and the combined organic phases were washed with saturated brine (15mL) and dried over anhydrous sodium sulfate. Filtration and concentration under reduced pressure, and the residue was purified by column chromatography (petroleum ether: ethyl acetate 1:1) to give the product 8.8(30mg, yellow solid, yield: 31%). LCMS ESI (+) M/z 567.1(M +1).

Step I:1- (5, 11-dihydrobenzo [6',7' ] thiaheptacyclo [3',4': 4,5] benzo [1,2-c ]

[1,2,5] Thiadiazol-11-yl-5-hydroxy-3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione (example 8)

Compound 8.8(30mg, 0.05mmol) was dissolved in 2mL of N, N-dimethylformamide, and lithium chloride (200mg, 4.76mmol) was added. The reaction solution was stirred at 100 ℃ under nitrogen for 3 hours. Concentrating the reaction solution under reduced pressure, and purifying the residue by reverse phase preparation

Example 8(4mg, yellow solid, yield: 17%). LCMS ESI (+) M/z 477.1(M +1).

Example 9: (2147)

1- (6, 12-Dihydrobenzo [6',7' ] thiaheptacyclo [3',4':3,4] benzo [1,2-C ] [1,2,5] oxadiazol-6-yl) -5-hydroxy-3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] oxazine-4, 6-dione

The specific reaction equation is as follows:

step A: 4-amino-2-methyl-3-nitrobenzoic acid (Compound 9.1)

4-acetylamino-2-methylbenzoic acid (2.5g, 13.0mmol) was slowly dissolved in 18mL of concentrated sulfuric acid at room temperature, and a mixture of concentrated nitric acid (0.55mL) and concentrated sulfuric acid (1.3mL) was added dropwise to the reaction mixture in an ice bath, followed by stirring at room temperature overnight. LCMS monitored the starting material reaction completion. Water (40mL) was added to the reaction solution and a yellow solid precipitated, which was filtered and dried to give crude product 9.1(1.12g, yield: 44.8%, yellow solid). LCMS ESI (-) M/z 195.1(M-1).

And B: 6-carboxy-7-methylbenzo [ c ] [1,2,5] oxadiazole 1-oxide (Compound 9.2)

Crude starting material 9.1(1.12g, 5.7mmol) was dissolved in ethanol (15mL) and potassium hydroxide (638mg, 5.7mmol), water (1mL) was added to the reaction flask. 7.5% sodium hypochlorite (10.4g, 10.6mmol) was slowly added dropwise to the reaction mixture under ice-bath conditions, the reaction was stirred at room temperature for 0.5 hour, LCMS was used to monitor completion of the reaction of the starting material, the reaction mixture was concentrated under reduced pressure, water (35mL) was added to dissolve the residue, the residue was washed with ethyl acetate (15mL) 2 times, and the organic phase was discarded. The aqueous phase was acidified (pH 4) with hydrochloric acid and extracted 3 times with ethyl acetate (15 mL). The combined organic phases were dried over anhydrous sodium sulfate. Filtration, spin-drying and purification of the residue on a silica gel column (petroleum ether: ethyl acetate: 2:1) gave the desired product 9.2(650mg, dark yellow solid, yield: 59.1%). LCMS ESI (-) M/z 193.0 (M-1).

And C: 4-methylbenzo [ c ] [1,2,5] oxadiazole-5-carboxylic acid (Compound 9.3)

Compound 9.2(650mg, 3.4mmol) was dissolved in 10mL ethanol, then triphenylphosphine (1.75g, 6.7mmol) was added to the reaction solution, and the reaction was heated to 85 ℃ and stirred for 2 hours under a nitrogen atmosphere. LC-MS monitored the completion of the reaction of the starting materials. After concentration under reduced pressure, the residue was taken up in water (25mL), the aqueous phase was adjusted to pH 9 with dilute sodium hydroxide solution, washed 2 times with ethyl acetate (15mL) and the ethyl acetate phase discarded. The aqueous phase was adjusted to pH 4 with hydrochloric acid, extracted 3 times with ethyl acetate (15mL), and the combined organic solutions were dried over anhydrous sodium sulfate. Filtration and spin-drying gave the desired product 9.3(275mg, yield: 46.1%). LCMS ESI (-) M/z 177.0 (M-1).

Step D: 4-methylbenzo [ c ] [1,2,5] oxadiazole-5-carboxylic acid methyl ester (Compound 9.4)

Compound 9.3(275mg, 1.5mmol) was dissolved in 6mL of N, N-dimethylformamide, sodium hydride (92.4mg, 2.3mmol) was added slowly at zero temperature, and stirred for 30 minutes, and methyl iodide (263mg, 1.9mmol) was slowly added dropwise and stirred for 30 minutes at room temperature. The completion of the feed reaction was monitored by TLC. The reaction was quenched by addition of saturated ammonium chloride solution (3mL), extracted 2 times with ethyl acetate (15mL), the combined organic phases were dried over anhydrous sodium sulfate, filtered, spun-dried, and purified by silica gel column (petroleum ether: ethyl acetate ═ 6:1) to give the desired product 9.4(210mg, yield: 70.8%).

Step E: 4- (bromomethyl) benzo [ c ] [1,2,5] oxadiazole-5-carboxylic acid methyl ester (Compound 9.5)

Starting material 9.4(210mg, 1.14mmol) was dissolved in carbon tetrachloride (5mL) and then N-bromosuccinimide (223mg, 1.25mmol) and benzoyl peroxide (11mg, 0.05mmol) were added sequentially. The mixture was heated to 85 ℃ under nitrogen protection for reaction overnight. After completion of the reaction, the reaction mixture was filtered through celite, the cake was washed 2 times with carbon tetrachloride (12mL), and the filtrate was washed with 10% sodium hydrogensulfite (15mLX2) and saturated brine (20mL) in this order. After drying, the crude product 9.5 was obtained (220mg, pale yellow oil, yield: 74.3%).

Step F: 4- ((phenylsulfanyl) methyl) benzo [ c ] [1,2,5] oxadiazole-5-carboxylic acid methyl ester (Compound 9.6)

Compound 9.5(220mg, 0.81mmol) was weighed out and dissolved in 6mL of N, N-dimethylformamide, cooled to 0 deg.C, added with potassium carbonate (224mg, 1.62mmol) and thiophenol (89mg, 0.81mmol), and stirred for 1 hour while cooling on ice. Completion of the reaction was monitored by TLC. Water (25mL) was added to the reaction flask, extracted 3 times with ethyl acetate (12mL) and the organic phase was washed with saturated brine (20 mL). Dried over anhydrous sodium sulfate. Filtration and concentration under reduced pressure gave the desired product 9.6(200mg, pale yellow oil, yield: 90.5%).

Step G: 4- ((phenylthio) methyl) benzo [ c ] [1,2,5] oxadiazole-5-carboxylic acid (Compound 9.7)

Compound 9.6(200mg, 0.67mmol) was dissolved in 5mL of tetrahydrofuran and 1mL of methanol, and sodium hydroxide solution (2mL, 4M) was added and stirred at 80 ℃ for 1 hour. After completion of the reaction, most of the organic solvent was removed by concentration, and the residue was dissolved in 25mL of water, adjusted to pH 3 with hydrochloric acid, and extracted with ethyl acetate (15mLX 3). Dried over anhydrous sodium sulfate. Filtration and spin-drying gave crude product 9.7(185mg, brown oil, yield: 97%). LCMS ESI (-) M/z 258.0(M-1).

Step H: benzo [6',7' ] thiaheptacyclo [3',4':3,4] benzo [1,2-C ] [1,2,5] oxadiazol-6 (12H) -one (Compound 9.8)

Compound 9.7(185mg, 0.65mmol) was dissolved in 5mL of polyphosphoric acid, heated to 120 ℃ and stirred for 4 hours. TLC monitored the reaction was complete. Water (25mL) was added to the reaction mixture, which was extracted with ethyl acetate (15mLX3) and washed with sodium bicarbonate solution (20 mL). Drying the anhydrous sodium sulfate. Filtration and spin-drying gave crude 9.8(65mg, yield: 37.6%, yellow solid).

Step H: benzo [6',7' ] thiaheptacyclo [3',4':3,4] benzo [1,2-C ] [1,2,5] oxadiazol-6 (12H) -ol (Compound 9.9)

Compound 9.8(65mg, 0.24mmol) was dissolved in 5mL of tetrahydrofuran and 1mL of methanol under ice-bath conditions, sodium borohydride (9.2mg, 0.24mmol) was added, and stirring was continued at room temperature for 0.5 h. The completion of the starting reaction was monitored by TLC. The reaction was quenched by the addition of saturated ammonium chloride solution (2.5mL), extracted with ethyl acetate (12mLX2), and washed with saturated brine (20 mL). Dried over anhydrous sodium sulfate. Filtration and concentration gave the desired product 9.9(64mg, yellow solid, yield: 97.7%).

Step I: 5- (benzyloxy) -1- (6, 12-dihydrobenzo [6',7' ] thiaheptacyclo [3',4':3,4] benzo [1,2-C ] [1,2,5] oxadiazol-6-yl) -3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione (Compound 9.10)

Intermediate A (50mg, 0.168mmol) was dissolved in 3.5mL 1-propylphosphoric anhydride (50% in DMF), Compound 9.9(54.4mg, 0.2mmol) was added, and the reaction was heated to 100 ℃ under nitrogen and stirred for 1.5 h. LCMS monitored the starting material reaction complete. The reaction was quenched with saturated sodium bicarbonate solution (5mL), and the aqueous phase was extracted with ethyl acetate (15mLX2) and dried over anhydrous sodium sulfate. Filtration, spin-drying and purification by column chromatography (dichloromethane: methanol 10:1) gave the desired product 9.10(30mg, yield: 32.6%, light yellow oil). LCMS ESI (+) M/z 550.2(M +1).

J: 1- (6, 12-Dihydrobenzo [6',7' ] thiaheptacyclo [3',4':3,4] benzo [1,2-C ] [1,2,5] oxadiazol-6-yl) -5-hydroxy-3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione (example 9)

Compound 9.10(30mg, 0.06mmol) was dissolved in 5mL of N, N-dimethylformamide, and lithium chloride (231mg, 5.5mmol) was added to the reaction solution, which was then heated at 100 ℃ and stirred for 5 hours. LCMS monitored the starting material reaction complete. The reaction solution was directly spun dry, and the residue was purified by reverse preparation to give example 9(3.5mg, yield: 14%, pale yellow solid). LCMS ESI (+) M/z 460.1(M +1).

Example 10 (2111)

5-hydroxy-3, 3-dimethyl-1- (4-oxo-4, 6,7, 12-tetrahydrobenzo [ d ] pyrido [1,2-a ] azepin-12-yl) -2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione

The specific reaction equation is as follows:

step A: 6-Hydroxypyridinecarboxylic acid methyl ester (Compound 10.1)

6-Hydroxypicolinic acid (5g,36mmol) was dissolved in 10mL of methanol, 30mL of methanol hydrochloride solution (4M) was added, and the mixture was stirred at 40 ℃ overnight. The reaction solution was spin-dried to obtain 10.1(5g, yield: 90%). LCMS ESI (+) M/z:154.1(M +1).

And B: 6-oxo-1-phenethyl-1, 6-dihydropyridine-2-carboxylic acid methyl ester (Compound 10.2)

Compound 10.1(1.5g,9.8mmol) was dissolved in 50mL of toluene and 0.5mL of water, and (2-bromoethyl) benzene (2.72g,15mmol), potassium carbonate (2.7g,19.6), lithium bromide (2.13g,24.5mmol) and tetrabutylammonium bromide (316mg,0.98mmol) were added and stirred at 100 deg.C overnight. To the reaction mixture was added 150mL of water, followed by extraction with ethyl acetate (50mLX3), and the organic phases were combined, washed with saturated brine (150mL) and dried over anhydrous sodium sulfate. Filtration, spin-drying and column purification gave 10.2(100mg, yield: 4%). LCMS ESI (+) M/z:258.1(M +1).

And C: 6-oxo-1-phenethyl-1, 6-dihydropyridine-2-carboxylic acid ((Compound 10.3)

Compound 10.2(100mg, 0.39mmol) was dissolved in 5mL of methanol solution, and 3mL of aqueous sodium hydroxide (6M) was added, heated to 90 ℃ and stirred for 1 hour. The reaction solution was adjusted to pH 4 by adding hydrochloric acid solution. Extracted three times with ethyl acetate (15mL), the organic phases were combined, washed with saturated brine, and the organic phase was dried over anhydrous sodium sulfate. Filtration and spin-drying gave 10.3(80mg, yield: 84.6%). LCMS ESI (+) M/z 244.1(M +1).

Step D: 6, 7-dihydrobenzo [ d ] pyrido [1,2-a ] aza-4, 12-dione (Compound 10.4)

Compound 10.3(80mg,0.33mmol) was added to 5mL of polyphosphoric acid, and the reaction was stirred at 120 ℃ for 2 hours. After the reaction liquid is cooled to room temperature, sodium bicarbonate aqueous solution is slowly added to quench the reaction. Extraction was performed with ethyl acetate (30mLX3), and the organic phases were combined, washed with 50mL of saturated brine, and dried over anhydrous sodium sulfate. Filtration, spin-drying and column purification gave 10.4(50mg, yield: 67.8%). LCMS ESI (+) M/z:226.1(M +1).

Step E: 12-hydroxy-6, 7-dihydrobenzo [ d ] pyrido [1,2-a ] aza-4 (12H) -one (Compound 10.5)

Compound 10.4(50mg, 0.22mmol) was dissolved in 2mL tetrahydrofuran, and sodium borohydride (10mg,0.26mmol) was added at 0 deg.C, warmed to room temperature and stirred for 30 minutes. The reaction was quenched by addition of aqueous ammonium chloride solution, extracted with ethyl acetate (5mLX3), the organic phases were combined, washed with saturated brine (50mL), and the organic phase was dried over anhydrous sodium sulfate. Filtration and spin-drying gave 10.5(47mg, yield: 93%). LCMS ESI (+) M/z 221(M +1).

Step F:5- (benzyloxy) -3, 3-dimethyl-1- (4-oxo-4, 6,7, 12-tetrahydrobenzo [ d ] pyrido [1,2-a ] azepin-12-yl) -2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione (Compound 10.6)

Intermediate A (60mg,0.2mmol) was dissolved in 2mL of N, N-dimethylformamide, and Compound 10.5(48mg, 0.21mmol) and 3mL of 1-propylphosphoric anhydride solution (50% in DMF) were added to the reaction solution, which was stirred at 100 ℃ for 1 hour. The reaction was quenched with saturated sodium bicarbonate solution (5mL), and the aqueous phase was extracted with dichloromethane (15mL) and dried over anhydrous sodium sulfate. Filtration, spin-drying and purification gave 10.6(8mg, yield: 7.5%). LCMS ESI (+) M/z:508.2(M +1)

Step G: 5-hydroxy-3, 3-dimethyl-1- (4-oxo-4, 6,7, 12-tetrahydrobenzo [ d ] pyrido [1,2-a ] azepin-12-yl) -2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione (example 10)

Compound 10.6(8mg, 0.02mmol) was dissolved in 2mL of methanol and a catalytic amount of palladium on carbon was added under hydrogen protection. After stirring at room temperature for 30 minutes, the reaction mixture was filtered, spun-dried and prepared by reverse phase to give the product, example 10(3mg, yield: 45.6%). LCMS ESI (+) M/z 418.2(M +1).

Example 11(2139)

1- (5, 10-dihydrobenzo [5,6] thiaheptacyclo [2,3-b ] pyridin-5-yl) -5-hydroxy-3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,22-b ] pyridazine-4, 6-dione

The specific reaction equation is as follows:

step A methyl 2- (benzylthio) nicotinate (Compound 11.1)

The compound methyl 2-fluoronicotinate (1g, 6.5mmol) was dissolved in 10mL of N, N-dimethylformamide, and benzylthiol (850mg, 7.74mmol) and potassium carbonate (1.8g, 12.9mmol) were added at room temperature, followed by stirring for 2 hours. The reaction was quenched with 20mL of water, extracted with ethyl acetate (30mLX3), the organic phases were combined, washed with saturated brine and dried over anhydrous sodium sulfate. Filtration, spin-drying and purification on silica gel column gave the product 11.1(800mg, yield: 48%). LCMS ESI (+) M/z:260.1(M +1).

And B:2- (benzylthio) nicotinic acid (Compound 11.2)

Compound 11.1(800mg, 3.1mmol) was dissolved in 15mL of methanol, 5mL of sodium hydroxide solution (4M) was added, and the mixture was stirred at room temperature for 8 hours. The reaction solution was directly spin-dried and purified to obtain crude product 11.2(800mg, yield: 93%). LCMS ESI (+) M/z 246.1 (M +1).

And C: benzo [5,6] thiaheptacyclo [2,3-b ] pyridin-5 (10H) -one (Compound 11.3)

A solution of compound 11.2(800mg, 3.3mmol) was added to 5mL of polyphosphoric acid, heated to 120 ℃ and stirred for 2 hours. After cooling to room temperature, it was diluted with 20mL of water, extracted with ethyl acetate (30mLX3), the organic phases were combined, washed with a saturated solution of sodium hydrogencarbonate and saturated brine, dried over anhydrous sodium sulfate, filtered, spun-dried, and purified by column chromatography to give the product 11.3(50mg, yield: 7%). LCMS ESI (+) M/z 228.0(M +1).

Step D:5, 10-dihydro-benzo [5,6] thiepino [2,3-b ] pyridin-5-ol (Compound 11.4)

Compound 11.3(50mg, 0.22mmol) was dissolved in 5mL of tetrahydrofuran solution, sodium borohydride (17mg, 0.44mmol) was added portionwise at 0 deg.C, the reaction was stirred at room temperature for 30 minutes, quenched with saturated ammonium chloride solution, extracted with ethyl acetate (20mLX3), the organic phases were combined, washed with saturated brine, and dried over anhydrous sodium sulfate. Filtration, spin-drying and purification on silica gel column gave 11.4(45mg, yield: 91%). LCMS ESI (+) M/z:230.1(M +1).

Step E:5- (benzyloxy) -1- (5, 10-dihydrobenzo [5,6] thiepino [2,3-b ] pyridin-5-yl) 1,2-b ] pyridazine-4, 6-dione (Compound 11.5)

Intermediate A (45mg, 0.20mmol) and compound 11.4(45mg, 0.20mmol) were dissolved in 5mL of a solution of 1-propylphosphoric anhydride (50% in EA), heated to 100 deg.C, and stirred for 1 hour. After the reaction mixture was cooled, it was diluted with 10mL of water, extracted with ethyl acetate (20mLX3), and the organic phases were combined, washed with saturated brine and dried over anhydrous sodium sulfate. Filtration, spin-drying and purification on silica gel column gave 11.5(15mg, yield: 15%). LCMS ESI (+) M/z 510.2(M +1).

Step F: 1- (5, 10-Dihydrobenzo [5,6] thiepino [2,3-b ] pyridin-5-yl) -5-hydroxy-3, 3-dimethyl-2, 3-dihydro-1H-pyrido [2-b ] pyridazine-4, 6-dione (example 11)

Compound 11.5(15mg, 0.03mmol) was dissolved in 3mL of N, N-dimethylformamide, lithium chloride (60mg, 1.4mmol) was added, and the mixture was stirred at 100 ℃ for 30 minutes. The reaction solution was directly spun dry, and the residue was dissolved in 3mL of methanol. Filtration and reverse phase preparation of the filtrate gave the product, example 11(6mg, yield: 48%). LCMS ESI (+) M/z:420.1(M +1).

Example 12(2102)

5-hydroxy-1- (2-methoxy-5, 11-dihydrobenzo [6,7] thiaheptacyclo [3,4-b ] pyridin-5-yl) -3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione

The specific reaction equation is as follows:

step A:2- (bromomethyl) -6-methoxypyridine (Compound 12.1)

The compound 2-methoxy-6-methylpyridine (2g, 16.3mmol) was dissolved in 30mL of carbon tetrachloride, N-bromosuccinimide (3.18g, 17.9mmol) and benzoyl peroxide (118mg, 0.5mmol) were added, and the mixture was reacted at 85 ℃ under nitrogen overnight. After completion of the reaction, the reaction mixture was filtered through celite, the filter cake was washed with carbon tetrachloride (15mLX2), and the filtrate was washed with 10% sodium hydrogen sulfite (20mLX2) and saturated brine (20mL) in this order. Dried over anhydrous sodium sulfate. After filtration and spin-drying, the desired product 12.1(1.68g, yield: 51.2%) was obtained.

And B: methyl 2- ((6-methoxypyridin-2-yl) thio) benzoate (Compound 12.2)

Compound 12.1(1.68g, 8.3mmol) was dissolved in 15mL of N, N-dimethylformamide under ice bath, followed by addition of potassium carbonate (2.3g, 16.6mmol) and methyl 2-mercaptobenzoate (1.68g, 10.0mmol), and stirring was continued for 1 hour under ice bath. Completion of the reaction was monitored by TLC. Water (45mL) was added to the reaction mixture, which was extracted with ethyl acetate (20mLX3), washed with saturated brine (30mL), and dried over anhydrous sodium sulfate. Filtration, spin-drying and purification on silica gel column gave 12.2(2.2g, yield: 91.7%). LCMS ESI (+) M/z 290.1(M +1).

And C:2- (((6-methoxypyridin-2-yl) methyl) thio) benzoic acid (Compound 12.3)

Compound 12.2(2.2g, 7.6mmol) was dissolved in 15mL of tetrahydrofuran and 5mL of methanol, followed by addition of 4mL of sodium hydroxide solution (4M) and stirring at 80 ℃ for 1 hour. After completion of the reaction, the reaction mixture was concentrated under reduced pressure, the residue was dissolved in water (30mL), the reaction mixture was adjusted to pH 3 with hydrochloric acid, and the aqueous phase was extracted with ethyl acetate (20mLX 3). The organic phases were combined and dried over anhydrous sodium sulfate. After filtration and spin-drying, the desired product 12.3(2g, yield: 95.6%, white solid) was obtained. LCMS ESI (-) M/z 274.1 (M-1).

Step D:2- (((6-hydroxypyridin-2-yl) methyl) thio) benzoic acid (Compound 12.4)

Compound 12.3(1.1g, 4mmol) was weighed out and dissolved in 12mL of N, N-dimethylformamide, then lithium chloride (1g, 24mmol) and p-toluenesulfonic acid monohydrate (4.56g, 24mmol) were added in this order, and the reaction was stirred at 120 ℃ for 2.5 hours. After completion of the reaction, the reaction mixture was diluted with water (30mL), extracted with ethyl acetate (15mLX3), and washed with saturated brine (25 mL). The organic phase was dried over anhydrous sodium sulfate. Filtration and spin-drying gave 12.4(940mg, yield: 90%). LCMS ESI (-) M/z 260.1(M-1).

Step E: 2-hydroxybenzo [6,7] thiaheptacyclo [3,4-b ] pyridin-5 (11H) -one (Compound 12.5)

Compound 12.4(940mg, 3.6mmol) was dissolved in 5mL of polyphosphoric acid and the reaction was stirred for 4 hours with heating to 120 ℃. TLC monitored the reaction complete. After cooling to room temperature, the reaction mixture was diluted with water (30mL), extracted with ethyl acetate (15mLX3), and washed with saturated sodium bicarbonate solution (20 mL). The organic phase was dried over anhydrous sodium sulfate, filtered and rotary-dried to give crude 12.5(810mg, yield: 92.6%). LCMS ESI (+) M/z 244.0(M +1).

Step F: 2-Methoxybenzo [6,7] thiaheptacyclo [3,4-b ] pyridin-5 (11H) -one (Compound 12.6)

Compound 12.5(810mg, 3.33mmol) was dissolved in 10mL of N, N-dimethylformamide. Cesium carbonate (1.63g, 4.99mmol), lithium bromide (724mg, 8.33mmol) and methyl iodide (520mg, 3.66mmol) were added successively under ice bath and the reaction mixture was stirred for 1 hour. The reaction was monitored by TLC for completion, and the reaction solution was diluted with water (30mL) and extracted with ethyl acetate (15mLX 3). The organic phases were combined and dried over anhydrous sodium sulfate. Filtration and spin-drying followed by purification on silica gel column (petroleum ether: ethyl acetate: 5:1) gave the desired product 12.6(130mg, yield: 15.2%, white solid). LCMS ESI (+) M/z:258.1(M +1).

Step G: 2-methoxy-5, 11-dihydrobenzo [6,7] thiaheptacyclo [3,4-b ] pyridin-5-ol (Compound 12.7)

Compound 12.6(130mg, 0.51mmol) was dissolved in 5mL tetrahydrofuran and 1mL methanol under ice-bath conditions, sodium borohydride (29mg, 0.77mmol) was added, and stirring was continued at zero degrees for 0.5 h. The reaction was quenched by the addition of saturated ammonium chloride solution (2.5mL), extracted with ethyl acetate (15mLX2), and the combined organic phases were washed with saturated brine (20mL) and dried over anhydrous sodium sulfate. After filtration and concentration, the product was obtained as 12.7(130mg, yield: 99.2%, white solid). LCMS ESI (+) M/z:260.1(M +1).

Step H:5- (benzyloxy) -1- (2-methoxy-5, 11-dihydrobenzo [6,7] thiaheptacyclo [3,4-b ] pyridin-5-yl) -3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione (Compound 12.8)

Intermediate A (100mg, 0.34mmol) was dissolved in 8mL of 1, 2-dichloroethane, and Compound 12.7(130mg, 0.5mmol) and dichloroacetic acid (2 drops) were added and the reaction was stirred at 85 ℃ for 2 hours. The starting material reaction was monitored by LCMS for completion. The reaction was quenched with saturated sodium bicarbonate solution (5mL), and the aqueous phase was extracted with dichloromethane (15mL) and dried over anhydrous sodium sulfate. Filtration and spin-drying, and purification of the residue on a silica gel column (dichloromethane: methanol 10:1) gave the product 12.8(90mg, pale yellow oil, yield: 50%). LCMS ESI (+) M/z:540.2(M +1).

Step I: 5-hydroxy-1- (2-methoxy-5, 11-dihydrobenzo [6,7] thiaheptacyclo [3,4-b ] pyridin-5-yl) -3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione (example 12)

Compound 12.8(90mg, 0.17mmol) was dissolved in 5mL of N, N-dimethylformamide, lithium chloride (350mg, 8.34mmol) was added, and the reaction mixture was stirred at 100 ℃ for 5 hours. The reaction was monitored by LCMS to completion. The reaction solution was directly spun dry, and the residue was purified by reverse phase preparation to give the product, example 12(36mg, yield: 48%, pale yellow solid). LCMSESI (+) M/z 450.1(M +1).1H NMR (400MHz, DMSO) 7.86(d, J ═ 8.4Hz,1H),7.32(d, J ═ 7.5Hz,1H), 7.14-7.07 (M,3H),6.85(d, J ═ 2.4Hz, 2H),6.76(d, J ═ 8.4Hz,1H),5.73(d, J ═ 7.6Hz,2H),5.20(s,1H),3.89(s,3H),3.80(s,2H), 1.38(s,3H),1.08(s,3H).

Example 13: (2141)

1- (5, 11-dihydrobenzo [6,7] thiaheptacyclo [3,4-b ] pyridin-5-yl) -5-hydroxy-3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,22-b ] pyridazine-4, 6-dione

The specific reaction equation is as follows:

step A2-chlorobenzo [6,7] thiaheptacyclo [3,4-b ] pyridin-5 (11H) -one (Compound 13.1)

Compound 12.5(60mg, 0.25mmol) was dissolved in 10mL of N, N-dimethylformamide and dichlorosulfoxide (59.5mg, 0.5mmol) was slowly added dropwise at zero temperature. Then heated to 70 ℃ for 1 h. After cooling, water and ethyl acetate (10mL each) were added, the layers were separated with stirring, the aqueous layer was extracted with ethyl acetate (10mL X3), and the organic layers were combined, washed with saturated brine (30mL) and dried over anhydrous sodium sulfate. Filtration, spin-drying and purification on silica gel column gave 13.1(58mg, yield: 89.8%, yellow oil). LCMS ESI (+) m/z 262.0; 264.0(M +1).

And B: benzo [6,7] thiaheptacyclo [3,4-b ] pyridin-5 (11H) -one (Compound 13.2)

Compound 13.1(58mg, 0.22mmol) was dissolved in 10mL of methanol, Pd/C (5mg) was added, the reaction system was replaced three times with a hydrogen balloon, and the mixture was heated under reflux at 80 ℃ for 12 hours. Cooling, filtration over celite, spin-drying and purification on a silica gel column gave 13.2(45mg, yield: 93.8%, yellow oil). LCMS ESI (+) M/z 228.0(M +1).

And C:5- (benzyloxy) -3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione compound (13.3)

Compound 13.2(45mg, 0.20mmol) was dissolved in 10mL tetrahydrofuran solution, and sodium borohydride (15.2mg, 0.40mmol) was added under ice-bath to warm to room temperature for 1 h. Saturated ammonium chloride solution (10mL) and ethyl acetate (10mL) were added, and the mixture was stirred and allowed to stand for separation. The aqueous phase was extracted with ethyl acetate (10mL X3), and the organic phases were combined, washed with saturated brine (50mL) and dried over anhydrous sodium sulfate. Filtration and spin-drying were carried out, and the residue was purified by means of a silica gel column to give the product 13.3(42mg, yield: 92.7%, yellow solid). LCMS ESI (+) M/z:230.1(M +1).

Step D:5- (benzyloxy) -1- (5, 11-dihydrobenzo [6,7] thiaheptacyclo [3,4-b ] pyridin-5-yl) -3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione (Compound 13.4)

Intermediate A (50mg,0.17 mmol) and compound 13.3(45.9mg, 0.2mmol) were added to 5mL of 1-propylphosphoric anhydride (50% in DMF) and heated to 100 ℃ for 1 h. After cooling to room temperature, a saturated sodium bicarbonate solution (10mL) and methylene chloride (10mL) were added, and the mixture was stirred and allowed to stand for separation. The aqueous phase was extracted with dichloromethane (10mL X3), the organic phases were combined, washed with saturated brine (50mL) and dried over anhydrous sodium sulfate. Filtration and spin-drying of the residue gave 13.4(37mg, yield: 43.3%, yellow oil) as a product, which was purified by silica gel column, LCMS ESI (+) M/z:510.2(M +1).

Step E1- (5, 11-Dihydrobenzo [6,7] thiaheptacyclo [3,4-b ] pyridin-5-yl) -5-hydroxy-3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,22-b ] pyridazine-4, 6-dione (example 13)

Compound 13.4(37mg, 0.07mmol) was dissolved in 10mL of N, N-dimethylformamide, and lithium chloride (0.15g, 3.6mmol) was added and heated to 100 ℃ for reaction for 3 h. Water (10mL) was added for dilution, extracted with dichloromethane (10mL X3), and the organic layers were combined. The extract was washed with saturated brine (50mL) and dried over anhydrous sodium sulfate. Filtration, spin-drying, reverse phase preparative purification of the residue and lyophilization gave the product, example 13(11mg, yield: 36%, light yellow solid), LCMS ESI (+) M/z:420.2(M +1).

Example 14: (2143)

1- (2-chloro-5, 11-dihydrobenzo [6,7] thiaheptacyclo [3,4-b ] pyridin-5-yl) -5-hydroxy-3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione

The specific reaction equation is as follows:

step A: 2-chloro-5, 11-dihydrobenzo [6,7] thiaheptacyclo [3,4-b ] pyridin-5-ol (Compound 14.1)

Compound 13.1(310mg, 1.2mmol) was dissolved in 20mL of tetrahydrofuran, and sodium borohydride (91.2mg, 2.4mmol) was added under ice-bath, followed by stirring at room temperature for 1 h. The reaction was quenched by the addition of saturated ammonium chloride solution (20mL), the aqueous phase was extracted with ethyl acetate (20mL X3), and the organic layers were combined, washed with saturated brine (50mL) and dried over anhydrous sodium sulfate. Filtration and spin-drying the residue was purified by silica gel column to give the product 14.1(290mg, yield: 92.8%, yellow solid). LCMS ESI (+) M/z 264.0(M +1).

Step B5- (benzyloxy) -1- (2-chloro-5, 11-dihydrobenzo [6,7] thiaheptacyclo [3,4-B ] pyridin-5-yl) -3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-B ] pyridazine-4, 6-dione (Compound 14.2)

Intermediate A (50mg,0.17 mmol) and compound 14.1(52.7mg, 0.2mmol) were dissolved in 5mL of 1-propylphosphoric anhydride (50% in DMF), heated to 100 ℃ for 1h, saturated sodium bicarbonate solution (10mL) was added, extracted with dichloromethane (10mLX3), the organic phases combined, washed with saturated brine (30mL) and dried over anhydrous sodium sulfate. Filtration and spin-drying of the residue gave 14.2(40mg, yield: 43.9%, yellow oil) as a residue which was purified by silica gel column chromatography. LCMS ESI (+) M/z:543.2/545.2(M +1).

And C:1- (2-chloro-5, 11-dihydrobenzo [6,7] thiaheptacyclo [3,4-b ] pyridin-5-yl) -5-hydroxy-3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione (Compound 14)

Compound 14.2(40mg, 0.07mmol) was dissolved in 10mL of N, N-dimethylformamide, lithium chloride (0.16g, 3.7mmol) was added, and the mixture was heated to 100 ℃ and stirred for 3 hours. Cooled to room temperature, diluted with water (10mL), extracted with dichloromethane (10mL X3), and the combined organic layers were washed with saturated brine (30mL) and dried over anhydrous sodium sulfate. Filtration and spin-drying were carried out, and the residue was purified by reverse phase preparative and lyophilized to give the target product, example 14(15mg, yield: 45%, pale yellow solid). LCMS ESI (+) m/z 454.2.

Example 15(2142)

5-hydroxy-3, 3-dimethyl-1- (2-methyl-5, 11-dihydrobenzo [6,7] thiaheptacyclo [3,4-b ] pyridin-5-yl) -2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione

The specific reaction equation is as follows:

step A: 2-bromo-benzo [6,7] thiaheptacyclo [3,4-b ] pyridin-5 (11H) -one (Compound 15.1)

Compound 12.5(50mg, 0.20mmol) was dissolved in 10mL of N, N-dimethylformamide, and phosphorus tribromide (108mg, 0.40mmol) was slowly added under ice bath. Stirred at room temperature for 10 minutes, then heated to 110 ℃ and reacted for 2 h. After cooling, water (10mL) was added for dilution, extraction was performed with ethyl acetate (10mL X3), and the organic layers were combined, washed with saturated brine (30mL) and dried over anhydrous sodium sulfate. Filtration and spin-drying, and purification of the residue on a silica gel column gave 15.1(60mg, yield: 79.5%, yellow oil). LCMS ESI (+) m/z 305.9; 307.9(M +1).

Step B2-methylbenzo [6,7] thiaheptacyclo [3,4-B ] pyridin-5 (11H) -one (Compound 15.2)

Compound 15.1(60mg, 0.20mmol) was dissolved in 10mL of toluene, potassium phosphate (212mg, 1.0mmol), methylboronic acid (24mg, 0.40mmol), [1,1' -bis (diphenylphosphino) ferrocene ] dichloropalladium dichloromethane (49mg, 0.06mmol) were added in this order, the reaction was replaced 3 times with nitrogen, heated to 110 ℃ and stirred for 3 h. Water (10mL) was added for dilution, extraction was performed with ethyl acetate (10mL X3), and the organic layers were combined, washed with saturated brine (50mL) and dried over anhydrous sodium sulfate. Filtration and spin-drying were carried out, and the residue was purified by means of a silica gel column to give 15.2(30mg, yield: 63.4%, yellow oil). LCMS ESI (+) M/z:242.1(M +1).

Step C2-methyl-5, 11-dihydrobenzo [6,7] thiaheptacyclo [3,4-b ] pyridin-5-ol (Compound 15.3)

Compound 15.2(30mg, 0.12mmol) was dissolved in 10mL of tetrahydrofuran, and sodium borohydride (9.1mg, 0.24mmol) was added under ice-bath to react at room temperature for 1 h. Saturated ammonium chloride solution (10mL) was added, extraction was performed with ethyl acetate (10mL X3), and the organic layers were combined, washed with saturated brine (50mL), and dried over anhydrous sodium sulfate. Filtration, spin-drying and purification on silica gel column gave 15.3(26mg, yield: 86.1%, yellow solid). LCMS ESI (+) M/z 244.1(M +1).

Step D5- (benzyloxy) -3, 3-dimethyl-1- (2-methyl-5, 11-dihydrobenzo [6,7] thiaheptacyclo [3,4-b ] pyridin-5-yl) -2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione (Compound 15.4)

Intermediate A (50mg,0.17 mmol) and compound 15.3(48.7mg, 0.2mmol) were dissolved in 5mL 1-propylphosphoric anhydride (50% in DMF) and heated to 100 ℃ and stirred for 1 h. The reaction was cooled to room temperature, saturated sodium bicarbonate solution (10mL) was added, extraction was performed with dichloromethane (10mL X3), and the organic phases were combined, washed with saturated brine (30mL) and dried over anhydrous sodium sulfate. Filtration and spin-drying were carried out, and the residue was purified by means of a silica gel column to give the product 15.4(35mg, yield: 39.9%, yellow oil). LCMS ESI (+) M/z 524.2(M +1).

Step E5-hydroxy-3, 3-dimethyl-1- (2-methyl-5, 11-dihydrobenzo [6,7] thiaheptacyclo [3,4-b ] pyridin-5-yl) -2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione (example 15)

Compound 15.4(35mg, 0.07mmol) was dissolved in 10mL of N, N-dimethylformamide, and lithium chloride (0.14g, 3.4mmol) was added and heated to 100 ℃ for reaction for 3 h. Cooled to room temperature, water (10mL) was added, extracted with dichloromethane (10mL X3), and the organic phases were combined, washed with saturated brine (20mL) and dried over anhydrous sodium sulfate. Filtration, spin-drying, reverse phase preparative purification and freeze-drying gave the product, example 15(10mg, yield: 34.5%, light yellow solid). LCMS ESI (+) M/z:434.2(M +1)

Example 16(2176)

2,2, 2-trifluoro-N- (5- (5-hydroxy-3, 3-dimethyl-4, 6-dioxo-2, 3,4, 6-tetrahydro-1H-pyrido [1,2-B ] pyridazin-1-yl) -5, 11-dihydrobenzo [6,7] thiaheptacyclo [3,4-B ] pyridin-2-yl) acetamide

The specific reaction example is as follows:

step A2-aminobenzo [6,7] thiaheptacyclo [3,4-b ] pyridin-5 (11H) -one (Compound 16.1)

Compound 15.1(4.4g, 14.4mmol), cuprous oxide (0.1g, 0.7mmol), ethylene glycol (50mL), and aqueous ammonia (50mL) were charged into a 250mL single-neck flask. Heating to 150 ℃ under the protection of nitrogen, refluxing overnight, adding 100mL of water and 100mL of ethyl acetate after complete reaction, stirring, standing and demixing. The aqueous layer was extracted with ethyl acetate (100mL X3), and the organic layers were combined, washed with saturated brine (150mL X1), and dried over anhydrous sodium sulfate. Filtration and spin-drying on silica gel column gave 16.1 as a yellow solid (2.3g, 65.7% yield). LCMS ESI (+) M/z 243(M +1).

And B:2, 2, 2-trifluoro-N- (5-oxo-5, 11-dihydro-benzo [6,7] thiaheptacyclo [3,4-b ] pyridin-2-yl) acetamide (Compound 16.2)

Compound 16.1(300mg, 1.24mmol), trifluoroacetic anhydride (390.6mg, 1.86mmol), and dichloromethane (10mL) were added to a 50mL single-necked flask, stirred at room temperature for 1 hour, after completion of the reaction, 20mL of water was added for dilution, the aqueous layer was extracted with dichloromethane (20mLX3), the organic layers were combined, washed with saturated brine (50mL X1), and dried over anhydrous sodium sulfate. Filtration, spin-drying, and purification on silica gel column gave 16.32 as a yellow oil (380mg, 90.7% yield). LCMS ESI (+) M/z:339(M +1).

And C:2, 2, 2-trifluoro-N- (5-hydroxy-5, 11-dihydrobenzo [6,7] thiaheptacyclo [3,4-b ] pyridin-2-yl) acetamide (Compound 16.3)

Compound 16.2(380mg, 1.12mmol) was dissolved in 10mL of tetrahydrofuran, sodium borohydride (63.8mg, 1.68mmol) was slowly added under ice bath, the mixture was stirred at room temperature for 0.5 hour, after completion of the reaction, saturated ammonium chloride (20mL, X3) was added thereto, and the mixture was quenched, the aqueous layer was extracted with ethyl acetate (20mL, X3), and the organic layers were combined, washed with saturated brine (50mL, X1), and dried over anhydrous sodium sulfate. Filtration, spin-drying and purification on silica gel column gave the product 16.3 as a pale yellow solid (80mg, 20.9% yield). LCMS ESI (+) M/z:341(M +1).

Step D: n- (5- (5- (benzyloxy) -3, 3-dimethyl-4, 6-dioxo-2, 3,4, 6-tetrahydro-1H-pyrido [1,2-b ] pyridazin-1-yl) -5, 11-dihydrobenzo [6,7] thiaheptacyclo [3,4-b ] pyridin-2-yl) -2,2, 2-trifluoroacetamide (compound 16.4)

Intermediate A (40mg, 0.13mmol) and compound 16.3(45.6mg, 0.13mmol) were added to a 10mL solution of 1-propylphosphoric anhydride (50% in DMF) and heated to 100 ℃ for 2 h. The mixture was diluted with 10mL of a saturated sodium bicarbonate solution and 10mL of dichloromethane, stirred, and then allowed to stand for separation. The aqueous layer was extracted with dichloromethane (10mL X3), the organic layers were combined, washed with saturated brine (50mL), dried over anhydrous sodium sulfate, filtered, spun-dried, and purified by silica gel column to give 16.4 as a yellow oil (40mg, 48.0% yield). LCMS ESI (+) M/z 621.2 (M +1).

Step E: 2,2, 2-trifluoro-N- (5- (5-hydroxy-3, 3-dimethyl-4, 6-dioxo-2, 3,4, 6-tetrahydro-1H-pyrido [1,2-B ] pyridazin-1-yl) -5, 11-dihydrobenzo [6,7] thiaheptacyclo [3,4-B ] pyridin-2-yl) acetamide (example 16)

Dissolving compound 16.4(40mg, 0.064mmol) in N, N-dimethylformamide, adding lithium chloride (136mg, 3.2mmol), heating to 100 ℃ for reaction for 3h, cooling after the reaction is completed, adding 10mL of water and 10mL of dichloromethane, stirring, and standing for layering. The aqueous layer was extracted with dichloromethane (10mL X3) and the organic layers were combined. The mixture was washed with saturated brine (50mL X1), and the organic phase was dried over anhydrous sodium sulfate. Filtration, spin-drying, reverse phase preparative purification of the residue and lyophilization afforded product 16(10mg, light yellow solid). LCMS ESI (+) M/z 531.1(M +1).

Example 17(2177)

N- (5- (5-hydroxy-3, 3-dimethyl-4, 6-dioxo-2, 3,4, 6-tetrahydro-1H-pyrido [1,2-b ] pyridazin-1-yl) -5-, 11-dihydrobenzo [6,7] thiaheptacyclo [3,4-b ] pyridin-2-yl) acetamide

The specific reaction steps are as follows:

step A: 2-amino-5, 11-dihydrobenzo [6,7] thiaheptacyclo [3,4-b ] pyridin-5-ol (Compound 17.1)

Compound 16.1(200mg, 0.83mmol) was added to 10mL tetrahydrofuran, sodium borohydride (47.1mg, 1.24mmol) was added under ice-bath, the mixture was warmed to room temperature and stirred for 0.5 h, and after completion of the reaction, 20mL saturated ammonium chloride was added and quenched. Extraction was performed with ethyl acetate (20 mL. times.3), and the organic layers were combined, washed with saturated brine (50mL), and dried over anhydrous sodium sulfate. Filtration, spin-drying and purification on silica gel column gave 17.1 as a pale yellow solid (180mg, 89.1% yield). LCMS ESI (+) M/z 245.1(M +1).

And B: 2-Acetylamino-5, 11-dihydrobenzo [6,7] thieno [3,4-b ] pyridin-5-yl acetate (Compound 17.2)

Compound 17.1(180mg, 0.74mmol) and triethylamine (373.7mg, 3.7mmol) were dissolved in 10mL of dichloromethane, acetyl chloride (232.4mg, 2.96mmol) was added to the solution under ice bath, the mixture was stirred at room temperature for 1 hour, and after completion of the reaction, 20mL of water and 20mL of dichloromethane were added to dilute the solution, and the mixture was stirred and allowed to stand for separation. The aqueous layer was extracted with dichloromethane (20mLX3), and the combined organic layers were washed with saturated brine (50ml), dried over anhydrous sodium sulfate, filtered, spun-dried, and purified by silica gel column to give 17.2(165mg, yield 68.2%) as a yellow oil. LCMS ESI (+) M/z:329.1(M +1).

And C: n- (5- (5- (benzyloxy) -3, 3-dimethyl-4, 6-dioxo-2, 3,4, 6-tetrahydro-1H-pyrido [1,2-b ] pyridazin-1-yl) -5, 11-dihydrobenzo [6,7] thiaheptacyclo [3,4-b ] pyridin-2-yl) acetamide (Compound 17.3)

Intermediate A (30mg, 0.10mmol) and compound 17.2(33mg, 0.10mmol) were dissolved in 10mL of 1-propylphosphoric anhydride (50% in DMF) solution and heated to 100 ℃ for 2 h. After the reaction was complete, 10mL of saturated sodium bicarbonate solution was added to dilute the reaction. The mixture was extracted with dichloromethane (10 mL. times.3), and the organic layers were combined, washed with saturated brine (50mL), and dried over anhydrous sodium sulfate. Filtration, spin-drying and purification on silica gel column gave 17.3 as a yellow oil (20mg, yield 35.1%). LCMS ESI (+) M/z 553.2(M +1).

Step D: n- (5- (5-hydroxy-3, 3-dimethyl-4, 6-dioxo-2, 3,4, 6-tetrahydro-1H-pyrido [1,2-b ] pyridazin-1-yl) -5-, 11-dihydrobenzo [6,7] thiaheptacyclo [3,4-b ] pyridin-2-yl) acetamide (Compound 17)

Compound 17.3(20mg, 0.035mmol) was dissolved in 10mL of NN-dimethylformamide, lithium chloride (74.4mg, 1.75mmol) was added, the mixture was heated to 100 ℃ for reaction for 3 hours, 10mL of water was added for dilution, extraction was performed with methylene chloride (10mL X3), the organic layers were combined, washed with saturated brine (50mL X1), and dried over anhydrous sodium sulfate. Filtration, spin-drying, reverse-phase preparative purification, and lyophilization gave product 17(155mg, white solid). LCMS ESI (+) M/z 477.1(M +1).

Example 18(2179)

1- (2- (dimethylamino) -5, 11-dihydrobenzo [6,7] thiaheptacyclo [3,4-b ] pyridin-5-yl) -5-hydroxy-3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione

The specific synthesis method comprises the following steps:

step A:2- (dimethylamino) -5, 11-dihydrobenzo [6,7] thiaheptacyclo [3,4-b ] pyridin-5-ol (Compound 18.1)

Compound 16.1(100mg, 0.41mmol) and paraformaldehyde (200mg) were dissolved in 10mL of acetonitrile, and 1mL of acetic acid was added dropwise under ice-bath, followed by stirring for 5 minutes, sodium triacetoxyborohydride (260.8mg, 1.23mmol) was added, stirring was carried out at room temperature overnight, after completion of the reaction, 20mL of saturated ammonium chloride was added to quench the reaction, extraction was carried out with ethyl acetate (20mL X3), the organic layers were combined, washed with saturated brine (50mL X1), and dried over anhydrous sodium sulfate. Filtration, spin-drying and purification on silica gel column gave 18.1 as a pale yellow solid (40mg, yield 35.6%). LCMS ESI (+) M/z 273.1(M +1).

And B:5- (benzyloxy) -1- (2- (dimethylamino) -5, 11-dihydrobenzo [6,7] thiaheptacyclo [3,4-b ] pyridin-5-yl) -3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione (Compound 18.2)

Intermediate A (42mg, 0.14mmol) and compound 18.1(38.1mg, 0.14mmol) were added to a 10mL solution of 1-propylphosphoric anhydride (50% in DMF), heated to 100 ℃ for 2h reaction, 10mL saturated sodium bicarbonate solution and 10mL dichloromethane were added, the layers were stirred and separated, the aqueous layer was extracted with dichloromethane (10mL X3), the organic layers were combined, washed with saturated brine (50mL X1), and dried over anhydrous sodium sulfate. Filtration, spin-drying, and purification on silica gel column gave 18.2 as a yellow oil (16mg, 20.5% yield). LCMS ESI (+) M/z 553.2(M +1).

And C:1- (2- (dimethylamino) -5, 11-dihydrobenzo [6,7] thiaheptacyclo [3,4-b ] pyridin-5-yl) -5-hydroxy-3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione (Compound 18)

Compound 18.2(16mg, 0.029mmol) was added to 10mL of N, N-dimethylformamide, lithium chloride (61.2mg, 1.45mmol) was added, the mixture was heated to 100 ℃ and reacted for 3 hours, 10mL of water and 10mL of methylene chloride were added, the mixture was stirred and the layers were separated, the aqueous layer was extracted with methylene chloride (10mL X3), the organic layers were combined, washed with saturated brine (50mL X1), and dried over anhydrous sodium sulfate. Filtration, spin-drying, reverse phase preparative purification, and lyophilization gave product 18 (white solid, 1 mg). LCMS ESI (+) M/z:463.2(M +1).

Example 19(2188)

1- (2-fluoro-5, 11-dihydrobenzo [6,7] thiaheptacyclo [3,4-b ] pyridin-5-yl) -5-hydroxy-3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione

The specific reaction equation is as follows:

step A: 2-fluoro-benzo [6,7] thiaheptacyclo [3,4-b ] pyridin-5 (11H) -one (Compound 19.1)

Compound 16.1(200mg, 0.83mmol) was dissolved in 10mL of pyridine hydrofluoric acid, and sodium nitrite (62.8mg, 0.92mmol) was added thereto while cooling on ice, followed by stirring at room temperature for 2 hours. After completion of the reaction, 20mL of a saturated sodium bicarbonate solution was slowly added, and the mixture was extracted with ethyl acetate (20mL of X3), and the organic layers were combined, washed with saturated brine (50mL) and dried over anhydrous sodium sulfate. Filtration, spin-drying and purification on silica gel column gave 19.1 as a pale yellow oil (190mg, 94.1% yield). LCMS ESI (+) M/z 246(M +1).

Step B2-fluoro-5, 11-dihydrobenzo [6,7] thiaheptacyclo [3,4-B ] pyridin-5-ol (Compound 19.2)

Compound 19.1(190mg, 0.78mmol) was dissolved in 10mL of tetrahydrofuran, sodium borohydride (44.5mg, 1.17mmol) was added under ice-bath, the mixture was stirred at room temperature for 0.5 hour, 20mL of saturated ammonium chloride was added after completion of the reaction, extraction was performed with ethyl acetate (20mL X3), the organic layers were combined, the organic phase was washed with saturated brine (50mL), and dried over anhydrous sodium sulfate. Filtration, spin-drying and purification on silica gel column gave 19.2 as a pale yellow solid (170mg, 88.5% yield). LCMS ESI (+) M/z:248.1(M +1).

And C:5- (benzyloxy) -1- (2-fluoro-5, 11-dihydrobenzo [6,7] thiaheptacyclo [3,4-b ] pyridin-5-yl) -3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione (Compound 19.3)

Intermediate A (35mg, 0.12mmol) and compound 19.2(29.6mg, 0.12mmol) were added to a solution of 10mL 1-propylphosphoric anhydride (50% in DMF) and heated to 100 ℃ for 2 h. After completion of the reaction, the reaction was quenched by addition of 10mL of saturated sodium bicarbonate solution, extracted with dichloromethane (10mL X3), and the organic layers were combined, washed with saturated brine (50mL) and dried over anhydrous sodium sulfate. Filtration, spin-drying and purification on silica gel column gave 19.3 as a yellow oil (23mg, 37.1% yield). LCMS ESI (+) M/z 528.2(M +1).

Step D1- (2-fluoro-5, 11-dihydrobenzo [6,7] thiaheptacyclo [3,4-b ] pyridin-5-yl) -5-hydroxy-3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione (Compound 19)

Compound 19.3(23mg, 0.044mmol) was dissolved in 10mL of N, N-dimethylformamide, and lithium chloride (93.5mg, 2.2mmol) was added and heated to 100 ℃ for reaction for 3 hours. 10mL of water was added for dilution, extracted with dichloromethane (10mL X3), and the organic layers were combined. The organic phase was washed with saturated brine (50mL) and dried over anhydrous sodium sulfate. Filtration, spin-drying, reverse phase preparative purification and freeze-drying gave the product, example 19 (white solid, 9 mg). LCMS ESI (+) M/z 438.1(M +1).

Referring to the synthesis of example 19, intermediate a was exchanged for intermediate B, C, D, E, respectively. Two-step synthesis with compound 19.2 gave example (20-23).

Example 20(2195)

1' - (2-fluoro-5, 11-dihydrobenzo [6,7] thiaheptacyclo [3,4-b ] pyridin-5-yl) -5' -hydroxy-1 ', 2' -dihydrospiro [ cyclobutane 1,3' -pyrido [1,2-b ] pyridazine ] -4', 6' -dione

LCMS ESI(+)m/z:450.1(M+1).

Example 21(2196)

1' - (2-fluoro-5, 11-dihydrobenzo [6,7] thiaheptacyclo-b ] pyridin-5-yl) -5' -hydroxy-1 ', 2' -dihydrospiro [ cyclopentane 3' -pyrido [1,2-b ] pyridazine ] -4', 6' -dione

LCMS ESI(+)m/z:464.1(M+1).

Example 22(2197)

[1,2,4] triazine-6, 8-dione of (12AR) -12- (2-fluoro-5, 11-dihydrobenzo [6,7] thiaheptacyclo [3,4-b ] pyridin-5-yl) -7-hydroxy 3,4,12,12a tetrahyd-1H- [1,4] oxazino [3,4-C ] pyrido [2,1-f ]

LCMS ESI(+)m/z:467.1(M+1).

Example 23(2301)

[1,2,4] triazine-4, 6-dione of 1- (2-fluoro-5, 11-dihydrobenzo [6,7] thiaheptacyclo [3,4-b ] pyridin-5-yl) -5-hydroxy-3- ((R) -1,1, 1-trifluoropropylpyridin-2-yl) -2, 3-dihydro-1H-pyrido [2,1-f ]

LCMS ESI(+)m/z:507.1(M+1).

Example 24(2173)

1- (3-chloro-5, 11-dihydrobenzo [6,7] thieno [4,3-c ] pyridin-11-yl) -5-hydroxy-3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione

Step A4- (bromomethyl) -2-methoxypyridine (Compound 24.1)

(2-methoxypyridin-4-yl) methanol (500mg, 3.6mmol) was dissolved in 10mL of dichloromethane, PBr 3(2 mL) was added while cooling on ice, and stirred for 1 hour. After completion of the reaction, the solvent was spun off, the reaction mixture was neutralized with saturated aqueous sodium bicarbonate (15mL), extracted with ethyl acetate (10mLX3), and the organic phase was washed with saturated brine (20mL) and dried over anhydrous sodium sulfate. Filtration, spin-drying and column chromatography purification of the residue yielded compound 24.1 as a colorless and transparent liquid (650mg, 89%). LCMS ESI (+) M/z:202.1(M +1).

And B: methyl 2- (((2-methoxypyridin-4-yl) methyl) thio) benzoate (Compound 24.2)

Compound 24.1(650mg, 3.2mmol) was dissolved in 10mL of NN-dimethylformamide, and methyl 2-mercaptobenzoate (567mg, 3.4mmol) and potassium carbonate (662mg,4.8mmol) were added at room temperature, followed by stirring at room temperature for 2 hours. The reaction mixture was poured into 100mL of water, extracted three times with 30mL of ethyl acetate, and the organic phase was washed with saturated brine and dried over anhydrous sodium sulfate. Filtration, spin-drying and column chromatography of the residue afforded compound 24.2(900mg, 97.3%) as a colorless oil. LCMS ESI (+) M/z 290.1(M +1).

Step C2- (((2-methoxypyridin-4-yl) methyl) thio) benzoic acid (Compound 24.3)

Compound 24.2(900mg,3.1mmol) was dissolved in 10mL of tetrahydrofuran, and lithium hydroxide monohydrate (391mg,16.3mmol) was added thereto at room temperature, followed by stirring at room temperature for 18 hours. The reaction solution was neutralized with concentrated hydrochloric acid to pH 5, followed by extraction twice with 20mL of ethyl acetate, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered by spin-drying, and spin-dried to give 24.3(700mg, 82.1%) as a white solid. LCMS ESI (+) M/z 276.1(M +1).

Step D2- (((2-hydroxypyridin-4-yl) methyl) thio) benzoic acid (Compound 24.4)

Compound 24.3(700mg,2.5mmol) was dissolved in 20mL of a solution of N, N-dimethylformamide, and LiCl (662mg, 15.8mmol) and TsOH (2.6g,15.1mmol) were added at room temperature, followed by heating to 120 ℃ and stirring for 2 hours. After cooling to room temperature, the reaction mixture was poured into water, extracted with ethyl acetate (15mLX3), and the combined organic phases were washed with saturated brine and dried over anhydrous sodium sulfate. Filtration and spin-drying gave compound 24.4(750mg) as a yellow solid. LCMS ESI (+) M/z 262.1(M +1).

Step E3-hydroxybenzo [6,7] thiaheptacyclo [4,3-c ] pyridine 11(5H) -one (Compound 24.5A), 1-hydroxybenzo [6,7] thiaheptacyclo [4,3-c ] pyridine 11(5H) -one (Compound 24.5B)

Compound 24.4(400mg,1.53mmol) was dissolved in 3g of polyphosphoric acid and stirred for 18 hours at 120 ℃. The reaction mixture was diluted with 40mL of water, extracted with ethyl acetate (20mLX3), and the organic phase was washed with sodium bicarbonate (20mL) and dried over anhydrous sodium sulfate. Filtration, spin-drying and column chromatography of the residue afforded 24.5A as a white solid (56mg) and 24.5B as a white solid (164 mg).

LCMS ESI (+) M/z 244.1(M +1). (Compound 24.5A), LCMS ESI (+) M/z:244.1(M +1). (Compound 24.5)

Step F3-chlorobenzo [6,7] thieno [4,3-c ] pyridine 11(5H) -one (compound 24.6)

Compound 24.5A (72mg,0.3mmol) was dissolved in 2mL of phosphorus oxychloride and stirred at 120 ℃ for 12 hours. The solvent was spin dried and quenched by adding warm water. The mixture was extracted with ethyl acetate (10mLX2), and the organic phase was washed with saturated brine and dried over anhydrous sodium sulfate. Filtration, spin-drying and column chromatography of the residue afforded compound 24.6(68mg, 87.9%) as a white solid. LCMSISI (+) M/z 262.2(M +1).

Step G: 3-chloro-5, 11-dihydrobenzo [6,7] thio [4,3-c ] pyridin-11-ol (Compound 24.7)

Compound 24.6(70mg,0.27mmol) was dissolved in tetrahydrofuran/methanol (3mL/0.5mL) under ice-bath conditions, followed by addition of sodium borohydride (21mg,0.55mmol) and stirring continued at zero degrees for 1 hour. The mixture was quenched by addition of saturated ammonium chloride solution (5mL), extracted 2 times with ethyl acetate (15mLX2), and washed with saturated brine (20 mL). Dried over anhydrous sodium sulfate. Filtration and concentration gave compound 24.7(80mg, as a yellow solid). LCMS ESI (+) M/z 264.1(M +1).

Step H5- (benzyloxy) -1- (3-chloro-5, 11-dihydrobenzo [6,7] thio [4,3-c ] pyridin-11-yl) -3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione (Compound 24.8)

Intermediate A (80mg,0.27mmol) was dissolved in 2mL of N, N-dimethylformamide, then compound 24.7(75mg,0.28mmol) and 2mL of 1-propylphosphoric anhydride (50% in DMF) solution were added and the reaction was stirred at 100 ℃ for 1 hour. The reaction was quenched with saturated sodium bicarbonate solution (5mL), and the aqueous phase was extracted with additional ethyl acetate (15mL) 2 times and dried over anhydrous sodium sulfate. Filtration, spin-drying and purification gave compound 24.8(70mg, 45.5%) as a yellow solid. LCMS ESI (+) M/z:544.2(M +1)

Step I1- (3-chloro-5, 11-dihydrobenzo [6,7] thiaheptacyclo [4,3-c ] pyridin-11-yl) -5-hydroxy-3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione (example 24)

Compound 24.8(35mg,0.064mmol) was dissolved in 2mL of N, N-dimethylformamide, and lithium chloride (30mg, 0.71mmol) was added and heated to 100 ℃ for 1 h. After completion of the reaction, the reaction mixture was cooled, diluted with 10mL of water, extracted with ethyl acetate (10mLX3), the organic layers were combined, and the organic layer was washed with saturated brine (50mLX1) and dried over anhydrous sodium sulfate. Filtration, spin-drying and purification of the residue by reverse phase preparative lyophilization gave example 24(6.5mg, 22.2%) as a yellow solid. LCMS ESI (+) M/z 454.1(M +1).

Example 25(2174)

1- (1-chloro-5, 11-dihydrobenzo [6,7] thiaheptacyclo [4,3-c ] pyridin-11-yl) -5-hydroxy-3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione

The specific reaction equation is as follows:

step A1-chlorobenzo [6,7] thieno [4,3-c ] pyridine 11(5H) -one (Compound 25.1)

Compound 24.5B (60mg, 0.25mmol) was dissolved in 2mL of phosphorus oxychloride and stirred at 120 ℃ for 12 hours. The solvent was spin dried and quenched by adding warm water. The mixture was extracted with ethyl acetate (10mLX2), and the organic phase was washed with saturated brine and dried over anhydrous sodium sulfate. Filtration and spin-drying were carried out, and the residue was subjected to column chromatography to give compound 25.1(12mg, 18.6%) as a white solid. LCMS ESI (+) M/z 262.2(M +1).

And B: 1-chloro-5, 11-dihydrobenzo [6,7] thio [4,3-c ] pyridin-11-ol (Compound 25.2)

Compound 25.1(12mg,0.046mmol) was dissolved in tetrahydrofuran/methanol (2mL/0.5mL) under ice-bath conditions, followed by addition of sodium borohydride (4mg,0.11mmol) and stirring continued at zero degrees for 1 hour. After addition of saturated ammonium chloride solution (5mL), the mixture was quenched, extracted 2 times with ethyl acetate (15mL), washed with saturated brine (20mL), and dried over anhydrous sodium sulfate. Filtration and spin-drying gave compound 25.2(10mg) as a yellow solid. LCMS ESI (+) M/z 264.1(M +1).

Step C5- (benzyloxy) -1- (1-chloro-5, 11-dihydrobenzo [6,7] thio [4,3-C ] pyridin-11-yl) -3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione (Compound 25.3)

Intermediate A (18mg, 0.06mmol) was dissolved in 1mL of N, N-dimethylformamide, then compound 25.2(10mg) and 1mL of 1-propylphosphoric anhydride (50% in DMF) were added and the reaction was stirred at 100 ℃ for 1 hour. The reaction was quenched with saturated sodium bicarbonate solution (5mL), and the aqueous phase was extracted with additional ethyl acetate (15mL) 2 times and dried over anhydrous sodium sulfate. Filtration, spin-drying and purification yielded compound 25.3(2.7mg, yellow solid, 13.3%). LCMS ESI (+) M/z:544.2(M +1)

Step D1- (1-chloro-5, 11-dihydrobenzo [6,7] thio [4,3-c ] pyridin-11-yl) -5-hydroxy-3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione (Compound 25.4)

Compound 25.3(2.7mg, 0.005mmol) was dissolved in 2ml of N, N-dimethylformamide, lithium chloride (30mg, 0.71mmol) was added, the reaction was heated to 100 ℃ for 1 hour, 5ml of water and 5ml of ethyl acetate were added, the mixture was stirred and the layers were separated, the aqueous layer was extracted with ethyl acetate (5ml X3), the organic layers were combined, washed with saturated brine (5ml X1), and dried over anhydrous sodium sulfate. Filtration, spin-drying and purification of the residue by reverse phase preparative lyophilization gave example 25(0.5mg, 22.2%) as a yellow solid. LCMS ESI (+) M/z 454.1(M +1).

Example 26(2135)

1- (5, 11-dihydrobenzo [6,7] oxa [3,4-b ] pyridin-5-yl) -5-hydroxy-3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,22-b ] pyridazine-4, 6-dione

The specific reaction equation is as follows:

step A:2- (bromomethyl) nicotinic acid methyl ester (Compound 26.1)

Methyl 2-methylnicotinate (1g, 6.62mmol) was dissolved in 15mL of carbon tetrachloride, N-bromosuccinimide (1.4g, 7.86mmol) and benzoyl peroxide (30mg) were added, and the reaction was stirred at 85 ℃ under nitrogen overnight. The reaction mixture was filtered through Celite, the cake was washed with carbon tetrachloride (15mL), and the filtrate was washed with 10% sodium hydrogen sulfite (20mLX2) and saturated brine (20mL) in that order and dried over anhydrous sodium sulfate. Filtration and spin-drying gave the desired product 26.1(640mg, yield: 42%). LCMS ESI (+) M/z:230.1, 232.1(M +1)

And B:2- (Phenoxymethyl) nicotinic acid methyl ester (Compound 26.2)

Compound 26.1(640mg, 2.78mmol) was dissolved in 10mL of N, N-dimethylformamide. Potassium carbonate (2.3g, 16.6mmol) and phenol (311mg,2.34mmol) were added under ice bath, and stirring was continued for 4 hours under ice bath. Water (45mL) was added to the reaction flask, extracted with ethyl acetate (20mLX3) and washed with saturated brine (30 mL). Dried over anhydrous sodium sulfate. Filtration and spin-drying, and column chromatography of the residue yielded 26.2(500mg, yield: 73.9%). LCMS ESI (+) M/z 244.1(M +1).

And C:2- (Phenoxymethyl) nicotinic acid (Compound 26.3)

Compound 26.2(500mg, 2.05mmol) was dissolved in 5mL of tetrahydrofuran and 1mL of methanol, and 5mL of sodium hydroxide solution (4M) was added and stirred at room temperature overnight. The reaction mixture was concentrated to remove the organic solvent, diluted with water (30mL), adjusted to pH 3 with hydrochloric acid, extracted with ethyl acetate (20mLX3), and the combined organic phases were washed with saturated brine (30mL) and dried over anhydrous sodium sulfate. Filtration and purification gave 26.3(350mg, yield: 83.9%). LCMS ESI (-) M/z 230.1(M-1).

Step D: benzo [6,7] oxa [3,4-b ] pyridin-5 (11H) -one (Compound 26.4)

Compound 26.3(350mg, 1.53mmol) was dissolved in 5mL of polyphosphoric acid, heated to 120 ℃ and stirred for 4 hours. The reaction mixture was cooled to room temperature, and water (30mL) was added to dilute the reaction mixture. Extraction was performed with ethyl acetate (15mLX3), and the organic phases were combined, washed with sodium bicarbonate (20mL), and dried over anhydrous sodium sulfate. Filtration and spin-drying gave 26.4(40mg) as a crude product. LCMS ESI (+) M/z 212.1(M +1).

Step E:5, 11-dihydrobenzo [6,7] oxa [3,4-b ] pyridin-5-ol (Compound 26.5)

Compound 26.4(40mg, 0.19mmol) was dissolved in 3mL tetrahydrofuran and 1mL methanol under ice-bath, sodium borohydride (12mg,0.32mmol) was added, and stirring was continued at zero degrees for 0.5 h. Quenched by addition of saturated ammonium chloride solution (2.5mL) and extracted with ethyl acetate (15mLX 2). The organic phases were combined, washed with saturated brine (20mL) and dried over anhydrous sodium sulfate. Filtration and concentration gave the crude product 26.5(60 mg). LCMS ESI (+) M/z:214.1(M +1).

Step F:5- (benzyloxy) -1- (5, 11-dihydrobenzo [6,7] oxa [3,4-b ] pyridin-5-yl) -3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione (Compound 26.6)

Intermediate A (60mg,0.20mmol) was dissolved in 2mL of N, N-dimethylformamide, a solution of compound 26.5(60mg, 0.28mmol) and 3mL of 1-propylphosphoric anhydride (50% in DMF) was added to a reaction flask, and the reaction was stirred at 100 ℃ for 1 hour. The reaction was quenched with saturated sodium bicarbonate solution (5mL), the aqueous phase was extracted with dichloromethane (15mLX2), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and purified by rotary drying to give 26.6(50mg, yield: 36.2%). LCMS ESI (+) M/z:494.2(M +1).

Step G: 1- (5, 11-Dihydrobenzo [6,7] oxa [3,4-b ] pyridin-5-yl) -5-hydroxy-3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,22-b ] pyridazine-4, 6-dione (Compound 26)

Compound 26.6(50mg,0.1mmol) was dissolved in 3mL of N, N-dimethylformamide, lithium chloride (150mg, 3.5mmol) was added, and the reaction mixture was stirred at 100 ℃ for 1 hour. After the reaction was completed, the solvent was directly removed by concentration under reduced pressure, and the residue was dissolved in 2mL of methanol and purified by reverse phase preparation to obtain the objective product, example 26(8mg, yield: 19.6%). LCMS ESI (+) M/z:404.1(M +1).

Example 27(2124)

5-hydroxy-3, 3-dimethyl-1- (3-methyl-6, 12-dihydro-3H-benzo [6',7' ] thiaheptacyclo [3',4':3,4] benzo [1,2-d ] [1,2,3] triazol-6-yl) -2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione

The specific reaction equation is as follows:

step A: 4-bromo-3-methyl-2-nitroaniline (compound 27.1)

3-methyl-2-nitroaniline (5.3g, 34.9mmol) was dissolved in 45mL of acetic acid, and N-bromosuccinimide (6.8g, 38.4mmol) was added. The reaction mixture was heated to 110 ℃ and stirred for 2 hours under an inert nitrogen blanket. The reaction was monitored by LCMS for completion, and the reaction was concentrated under reduced pressure, diluted with water (60mL), extracted with ethyl acetate (30mLX3), and the organic phases were combined and dried over anhydrous sodium sulfate. After filtration and spin-drying, the crude product 27.1(5.3g, yield: 65.8%) was obtained. LCMS ESI (+) m/z 231.0; 233.0(M +1).

And B: 4-bromo-3-methylbenzene-1, 2-diamine (Compound 27.2)

Compound 27.1(5.3g, 22.9mmol) was dissolved in 80mL of ethanol, and then iron powder (7.7g, 137.6mmol), ammonium chloride (12.4g, 229.4mmol) and water (40mL) were added in that order. The reaction was heated to reflux and stirred under nitrogen for 1 hour. The completion of the starting reaction was monitored by TLC. The reaction was filtered through celite, the filter cake was washed with methanol (15mL), and the filtrate was spin dried to give crude product 27.2(5.5g, brown solid). LCMS ESI (+) m/z: 201.1; 203.1(M +1).

And C: 5-bromo-4-methyl-1H-benzo [ d ] [1,2,3] triazole (Compound 27.3)

Compound 27.2(2g, 9.95mmol) was dissolved in 90mL of acetic acid and 45mL of water, a solution of sodium nitrite (686mg, 9.95mmol) in water (10mL) was slowly added dropwise, and the mixture was stirred at ambient temperature for an additional 0.5 hour. The reaction was concentrated and the residue was taken up in saturated sodium bicarbonate solution (30mL) and extracted with ethyl acetate (15mLX 2). The organic phases were combined and dried over anhydrous sodium sulfate. Filtration and spin-drying gave crude compound 27.3(1.7g, yield: 80.9%). LCMSISI (+) m/z 212.0; 214.0(M +1).

Step D: 5-bromo-1, 4-dimethyl-1H-benzo [ d ] [1,2,3] triazole (Compound 27.4)

Compound 27.3(1.5g, 7.1mmol) was dissolved in 12mL of N, N-dimethylformamide while cooling on ice, sodium hydride (312mg, 7.81mmol) was added slowly in portions, and stirred for 30 minutes, methyl iodide (1.33g, 9.37mmol) was added, and stirring was continued for 30 minutes while warming to room temperature. TLC monitored the starting material reaction was complete. The reaction was quenched by the addition of saturated ammonium chloride solution (6mL), extracted with ethyl acetate (15mLX2), and the organic phases were combined and dried over anhydrous sodium sulfate. Filtration and spin-drying, and purification of the residue on a silica gel column gave 27.4(1.42g, yield: 88.7%, product as a mixture, 3 isomers in total). LCMS ESI (+) m/z: 226.1; 228.1(M +1).

Step E:1, 4-dimethyl-1H-benzo [ d ] [1,2,3] triazole-5-carboxylic acid methyl ester (Compound 27.5)

Compound 27.4(1.31g, 5.79mmol) was dissolved in a mixed solvent of 30mL of methanol and 10mL of N, N-dimethylformamide, and [1,1' -bis (diphenylphosphino) ferrocene ] dichloropalladium dichloromethane complex (946mg, 1.16mmol), triethylamine (12mL) and octacarbonyldicobalt (1.38g, 4.05mmol) were added, and the reaction was heated to 80 ℃ under nitrogen and stirred overnight. The reaction solution was filtered through celite, the filter cake was washed with methanol (10mL), the filtrate was spin-dried, and the residue was purified by a silica gel column. Finally, 27.5(755mg, yield: 63.6%, LCMS shows 3 UV peaks) was obtained. LCMS ESI (+) M/z 206.1(M +1).

Step F: 4- (bromomethyl) -1-methyl-1H-benzo [ d ] [1,2,3] triazole-5-carboxylic acid methyl ester (Compound 27.6)

Starting material 27.5(830mg, 4.04mmol) was dissolved in 15mL of carbon tetrachloride, N-bromosuccinimide (791mg, 4.45mmol) and benzoyl peroxide (29.3mg, 0.12mmol) were added sequentially, and the mixture was stirred at 85 ℃ overnight under nitrogen. The reaction solution was filtered through celite, the cake was washed with carbon tetrachloride (15mLX2), and the filtrate was washed with 10% sodium hydrogen sulfite (20mLX2) and saturated brine (20mL) in this order. Dried over anhydrous sodium sulfate. After filtration and spin-drying, the desired product 27.6(1.08g, yield: 93.9%) was obtained.

Step G: 1-methyl-4- ((phenylthio) methyl) -1H-benzo [ d ] [1,2,3] triazole-5-carboxylic acid methyl ester (Compound 27.7)

Compound 27.6(1.08g, 3.8mmol) was dissolved in 15mL of N, N-dimethylformamide. Potassium carbonate (1.05g, 7.6mmol) and thiophenol (501mg, 4.56mmol) were added under ice-bath, and stirring was maintained at zero degrees for 1 hour. The completion of the starting reaction was monitored by TLC. Water (35mL) was added for dilution, and the aqueous phase was extracted with ethyl acetate (20mLX 3). The organic phases were combined, washed with saturated brine (30mL) and dried over anhydrous sodium sulfate. Filtration and spin-drying, and purification of the residue by column chromatography gave 27.7(1.0g, yield: 89.9%, mixture). LCMS ESI (+) M/z: 314.1(M +1).

Step H: 1-methyl-4- ((phenylthio) methyl) -1H-benzo [ d ] [1,2,3] triazole-5-carboxylic acid (Compound 27.8)

Starting material 27.7(420mg, 1.34mmol) was dissolved in 10mL tetrahydrofuran and 2mL methanol, then 2mL sodium hydroxide solution (4M) was added and stirred at 80 ℃ for 1 hour. After completion of the reaction, the reaction mixture was spin-dried, the residue was dissolved in water (25mL), the solution was adjusted to pH 3 with hydrochloric acid, and extracted with ethyl acetate (15mLX 3). The combined organic phases were dried over anhydrous sodium sulfate. After filtration and spin-drying, the product was 27.8(980mg, yield: 97.3%, mixture). LCMS ESI (-) M/z 298.1 (M-1).

Step I: 3-methyl-3H-benzo [6',7' ] thiaheptacyclo [3',4':3,4] benzo [1,2-d ] [1,2,3] triazol-6- (12H) -one (Compound 27.9)

Compound 27.8(390mg, 1.3mmol) was dissolved in 5mL of polyphosphoric acid and the reaction was stirred at 120 ℃ for 4 hours. TLC monitored the starting material reaction was complete. After cooling to room temperature, the reaction mixture was diluted with water (25mL), extracted with ethyl acetate (15mLX3), and the organic phase was washed with saturated sodium bicarbonate solution (20mL) and dried over anhydrous sodium sulfate. Filtration and spin-drying gave the crude product (730mg, yield: 68.2%, mixture of the three isomers).

Step J: 3-methyl-6, 12-dihydro-3H-benzo [6',7' ] thiaheptacyclo [3',4':3,4] benzo [1,2-d ] [1,2,3] triazol-6-ol (Compound 27.10)

Compound 27.9(250mg, 0.89mmol) was dissolved in 5mL of tetrahydrofuran and 1mL of methanol, and sodium borohydride (40.6mg, 1.1mmol) was added under ice bath, followed by continued stirring at room temperature for 0.5 hour. TLC monitored the starting material reaction was complete. The reaction was quenched by the addition of saturated ammonium chloride solution (2.5mL) and extracted with ethyl acetate (15mLX 2). The combined organic phases were washed with saturated brine (20mL) and dried over anhydrous sodium sulfate. Filtration and concentration to column purification gave product 27.10A (240mg, white solid); 27.10B (400 mg); 27.10C: (78 mg). LCMS ESI (+) M/z 266.1(M-17).

Step K: 5- (benzyloxy) -3, 3-dimethyl-1- (3-methyl-6, 12-dihydro-3H-benzo [6',7' ] thiaheptacyclo [3',4':3,4] benzo [1,2,2-d ] [1,2,3] triazol-6-yl) -2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione (Compound 27.11)

Intermediate A (50mg, 0.168mmol) was dissolved in 5mL of 1, 2-dichloroethane, then compound 27.10A (62mg, 0.22mmol) and dichloroacetic acid (2 drops) were added to the reaction, and the reaction was stirred at 85 ℃ for 2 hours. LCMS monitored the starting material reaction complete. The reaction was quenched with saturated sodium bicarbonate solution (5mL) and the aqueous phase was extracted with dichloromethane (15mLx 2). The combined organic phases were dried over anhydrous sodium sulfate. Filtration and spin-drying were carried out, and the residue was purified by a silica gel column (dichloromethane: methanol 10:1) to obtain the objective product 27.11(65mg, yield: 68.7%, pale yellow oil). LCMS ESI (+) M/z 564.2(M +1).

Step L: 5-hydroxy-3, 3-dimethyl-1- (3-methyl-6, 12-dihydro-3H-benzo [6',7' ] thiaheptacyclo [3',4':3,4] benzo [1,2-d ] [1,2,3] triazol-6-yl) -2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione (example 27)

Compound 27.11(65mg, 0.12mmol) was dissolved in 5mL of N, N-dimethylformamide, lithium chloride (242mg, 5.75mmol) was added, and the reaction mixture was stirred at 100 ℃ for 5 hours. LCMS monitored the starting material reaction complete. The reaction solution was directly spin-dried, the residue was dissolved in methanol (3mL) and filtered, and the filtrate was purified by reverse phase preparation to obtain the objective product, example 27(30mg, yield: 54.8%, pale yellow solid) after lyophilization. LCMS ESI (+) M/z 474.2(M +1).1H NMR (400MHz, DMSO)7.74(d, J ═ 8.5Hz,1H),7.68(d, J ═ 8.6Hz,1H), 7.46(d, J ═ 7.5Hz,1H), 7.11-7.03 (M,2H),6.92(d, J ═ 7.5Hz,1H),6.85(t, J ═ 7.0Hz, 1H),5.79(t, J ═ 10.8Hz,2H),5.40(s,1H),4.76(d, J ═ 13.0Hz,1H),4.30(s,3H),3.67(d, J ═ 15.0, 1H),3.33(d, J ═ 15.0, 1H), 1H (s,3H), 37.06 (d, J ═ 15.0, 1H), 1H, 3.06 (s,3H).

Example 28(2123)

5-hydroxy-3, 3-dimethyl-1- (2-methyl-6, 12-dihydro-2H-benzo [6',7' ] thiaheptacyclo [3',4':3,4] benzo [1,2-d ] [1,2,3] triazol-6-yl) -2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione

The target product was obtained by two-step synthesis, replacing 27.10A in step K of example 27 with 27.10B. LCMSISI (+) M/z 474.2(M +1).

Example 29(2124)

5-hydroxy-3, 3-dimethyl-1- (1-methyl-6, 12-dihydro-1H-benzo [6',7' ] thiaheptacyclo [3',4':3,4] benzo [1,2-d ] [1,2,3] triazol-6-yl) -2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione

The target product was obtained by two-step synthesis by replacing 27.10A in step K of example 27 with 27.10C. LCMSISI (+) M/z 474.2(M +1).

Example 30(2182)

The specific reaction equation is as follows:

step A: (E) -N' -hydroxy-N- (5-oxo-5, 11-dihydro-benzo [6,7] thiaheptacyclo [3,4-b ] pyridin-2-yl) carboxamidine (Compound 30.1)

Compound 16.1(200mg, 0.83mmol), N, N-dimethylformamide dimethyl acetal (147.6mg, 1.24mmol) was dissolved in 10mL of isopropanol. Stirring at 85 ℃ for 1 hour, cooling to 60 ℃, adding hydroxylamine hydrochloride (86.2mg, 1.24mmol), stirring for 1 hour, adding 20mL of water and 20mL of ethyl acetate after the reaction is completed, stirring, and standing for layering. The aqueous layer was extracted with ethyl acetate (20mL X3), and the organic layers were combined, washed with saturated brine (50mL X1), and dried over anhydrous sodium sulfate. Filtration, spin-drying and purification on silica gel column gave 30.1 as a pale yellow solid (160mg, yield 67.8%). LCMS ESI (+) M/z 286.1(M +1).

And B: benzo [6,7] thiaheptacyclo [4,3-e ] [1,2,4] triazolo [1,5-a ] pyridin-6 (12H) -one (Compound 30.2)

Compound 30.1(160mg, 0.56mmol) was dissolved in 10mL of tetrahydrofuran, and trifluoroacetic anhydride (176.4mg, 0.84mmol) was added thereto and stirred at room temperature overnight. After completion of the reaction, 20mL of saturated sodium bicarbonate and 20mL of ethyl acetate were added, the mixture was stirred and the layers were separated, the aqueous layer was extracted with ethyl acetate (20mL X3), the organic layers were combined, washed with saturated brine (50mL), and dried over anhydrous sodium sulfate. Filtration, spin-drying and purification on silica gel column gave compound 30.2 as a yellow solid (95mg, yield 63.3%). LCMSISI (+) M/z 268.1(M +1).

And C: 6, 12-Dihydrobenzo [6,7] thiaheptacyclo [4,3-e ] [1,2,4] triazolo [1,5-a ] pyridin-6-ol (Compound 30.3)

Compound 30.2(95mg, 0.36mmol) was dissolved in 10mL of tetrahydrofuran, and sodium borohydride (20.5mg, 0.54mmol) was slowly added under ice-bath, followed by stirring at room temperature for 0.5 hour. After completion of the reaction, saturated ammonium chloride 20m l and 20mL of ethyl acetate were added thereto, the mixture was stirred and the layers were separated, and the aqueous layer was extracted with ethyl acetate (20 mL. times.3), and the organic layers were combined, washed with saturated brine (50mL) and dried over anhydrous sodium sulfate. Filtration, spin-drying and purification on silica gel column gave 30.3 as a pale yellow solid (85mg, 88.5% yield). LCMSISI (+) M/z:270.1(M +1).

Step D:5- (benzyloxy) -1- (6, 12-dihydrobenzo [6,7] thiaheptacyclo [4,3-e ] [1,2,4] triazolo [1,5-a ] pyridin-6-yl) -3-, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione (Compound 30.4)

Intermediate A (55mg, 0.18mmol) and compound 30.3(48.4mg, 0.18mmol) were dissolved in 10mL of 1-propylphosphoric anhydride (50% in DMF) and heated to 100 ℃ for 2 h. After the reaction was completed, it was cooled, and 10mL of saturated sodium bicarbonate solution and 10mL of dichloromethane were added, the mixture was stirred and the layers were separated, the aqueous layer was extracted with dichloromethane (10mL X3), the organic layers were combined, washed with saturated brine (30mL), dried over anhydrous sodium sulfate, filtered, and dried by spinning, and the residue was purified by a silica gel column to obtain 30.4(25mg, yield 24.8%) as a yellow oil. LCMS ESI (+) M/z 550.2(M +1).

Step E:1- (6, 12-Dihydrobenzo [6,7] thiaheptacyclo [4,3-e ] [1,2,4] triazolo [1,5-a ] pyridin-6-yl) -5-hydroxy-3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione (Compound 30)

Compound 30.4(25mg, 0.045mmol) was dissolved in 10mL of N, N-dimethylformamide, lithium chloride (93.5mg, 2.2mmol) was added, and the mixture was heated to 100 ℃ and reacted for 3 h. After completion of the reaction, it was cooled, 10mL of water and 10mL of dichloromethane were added, the layers were separated by stirring, the aqueous layer was extracted with 10mL of dichloromethane X3), the organic layers were combined, washed with saturated brine (50mL X1), dried over anhydrous sodium sulfate, filtered, spun-dried, purified by reverse phase preparative purification, and lyophilized to give example 30(10mg, white solid). LCMSISI (+) M/z 460.1(M +1).

Example 31(2170)

The specific reaction equation is as follows:

step A: pyrazolo [1,5-a ] pyridine-3-carboxylic acid ethyl ester (compound 31.1)

1-Aminopyridine iodide (2g, 9mmol) and ethyl propiolate (0.88g, 9mmol) were dissolved in 30mL of N, N-dimethylformamide, potassium carbonate (1.86g, 13.5mmol) was added thereto while cooling on ice, the mixture was stirred at room temperature overnight, after completion of the reaction, 50mL of water was added to dilute the mixture, the mixture was extracted with ethyl acetate (30mL of X3), the organic layers were combined, washed with saturated brine (100mL), and dried over anhydrous sodium sulfate. Filtration, spin-drying and purification on silica gel column gave 31.1 as a red oil (2g, 50.0% yield). LCMS ESI (+) M/z 191.1(M +1).

And B: pyrazolo [1,5-a ] pyridine-3-carboxylic acid (compound 31.2)

Compound 31.1(2g, 10.53mmol) was dissolved in 20mL of ethanol, 10mL of sodium hydroxide solution (4M) was added, the mixture was refluxed at 80 ℃ for 2 hours, and after completion of the reaction, the mixture was acidified with acid to precipitate a pink solid, which was then filtered, and the filtrate was washed with water 2 times and dried to obtain 31.2(1g, yield 58.8%) as a pink solid. LCMS ESI (-) M/z 161.0 (M-1).

And C: 3-Bromopyrazolo [1,5-a ] pyridine (Compound 31.3)

Compound 31.2(1g, 6.17mmol), N-bromosuccinimide (1.21g, 6.79mmol) and sodium hydrogencarbonate (1.55g, 18.51mmol) were added to 20mL of N, N-dimethylformamide, and stirred at room temperature overnight, after completion of the reaction, 20mL of water and 20mL of ethyl acetate were added, the mixture was stirred to separate layers, the aqueous layer was extracted with ethyl acetate (30mL X3), the organic layers were combined, washed with saturated brine (100mL) and dried over anhydrous sodium sulfate. Filtration, spin-drying and purification on silica gel column gave 31.3 as a yellow oil (0.8g, 65.6% yield). LCMS ESI (+) M/z:197.0/199.0(M +1).

Step D: methyl 2- (pyrazolo [1,5-a ] pyridin-3-ylthio) benzoate (Compound 31.4)

Compound 31.3(0.8g, 4.06mmol), methyl thiosalicylate (1.71g, 10.15mmol), tris (dibenzylideneacetone) dipalladium (742mg, 0.81mmol), 4, 5-bis (diphenylphosphino) -9, 9-dimethylxanthene (705mg, 1.22mmol) and N, N-diisopropylethylamine (1.57g, 12.18mmol) were added to dioxane, refluxed at 105 ℃ overnight under nitrogen protection, after completion of the reaction, 20mL of water and 20mL of ethyl acetate were added, the layers were separated by stirring, the aqueous layer was extracted with ethyl acetate (20mL of X3), the organic layers were combined, washed with saturated brine (100mL of X1), and dried over anhydrous sodium sulfate. Filtration, spin-drying, and purification on silica gel column gave 31.4 as a yellow oil (350mg, 30.4% yield). LCMS ESI (+) M/z 285.1(M +1).

Step E: 2- (pyrazolo [1,5-a ] pyridin-3-ylthio) benzoic acid (compound 31.5)

Compound 31.4(350mg, 1.23mmol) was dissolved in 10mL of methanol, 5mL of sodium hydroxide solution (4M) was added, the mixture was refluxed at 80 ℃ for 2 hours, and after completion of the reaction, the mixture was acidified with acid to precipitate a yellow solid, which was filtered, and the filtrate was washed with water 2 times and dried to obtain 31.5 as a pink solid (270mg, yield 81.1%). LCMS ESI (+) M/z 271.0(M +1).

Step F: 6H-11 thia 2, 2A-dibenzo [ cd, g ] azulen-6-one (Compound 31.6)

Compound 31.5(270mg, 1mmol) was added to 1g of polyphosphoric acid, stirred at 120 ℃ for 2 hours, after completion of the reaction, 20mL of water and 20mL of ethyl acetate were added, the mixture was stirred to separate layers, the aqueous layer was extracted with ethyl acetate (20mL X3), the organic layers were combined, washed with saturated brine (50mL X1), dried over anhydrous sodium sulfate, filtered, spun-dried, and purified by silica gel column to give 31.6(90mg, yield 35.7%) as a yellow oil. LCMS ESI (+) M/z:253.0(M +1).

Step G: 6H-11 thia 2, 2A-dibenzo [ cd, g ] azulen-6-ol (Compound 31.7)

Compound 31.6(90mg, 0.36mmol) was dissolved in 10mL of tetrahydrofuran, and sodium borohydride (20.5mg, 0.54mmol) was added under ice-bath, followed by stirring at room temperature for 0.5 hour. After completion of the reaction, 10mL of saturated ammonium chloride and 10mL of ethyl acetate were added, the mixture was stirred and the layers were separated, the aqueous layer was extracted with ethyl acetate (20mL X3), and the organic layers were combined, washed with saturated brine (20mL) and dried over anhydrous sodium sulfate. Filtration, spin-drying, and purification on silica gel column gave 31.7 as a pale yellow solid (86mg, yield 94.8%). LCMS ESI (+) M/z 237.0(M +1).

Step H: 1- (6H-11-Thiabe 2, 2A-dibenzo [ cd, g ] azulen-6-yl) -5- (benzyloxy) -3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,22-b ] pyridazine-4, 6-dione (Compound 31.8)

Intermediate A (56mg, 0.19mmol) and compound 31.7(48.3mg, 0.19mmol) were dissolved in 10mL of a solution of 1-propylphosphoric anhydride (50% in DMF), heated to 100 ℃ for 2h, added with 10mL of saturated sodium bicarbonate solution and 10mL of dichloromethane, stirred to separate layers, the aqueous layer was extracted with dichloromethane (10mL X3), the organic layers were combined, washed with saturated brine (25mL), dried over anhydrous sodium sulfate, filtered, spun-dried, and the residue was purified by silica gel column to give 31.8 as a yellow oil (45mg, 45.0% yield). LCMS ESI (+) M/z 535.2(M +1).

Step I:1- (6H-11-Thiaza 2, 2A-dibenzo [ cd, g ] azulen-6-yl) -5-hydroxy-3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2b ] pyridazine-4, 6-dione (Compound 31)

Compound 31.8(45mg, 0.084mmol) was dissolved in 10mL of N, N-dimethylformamide, lithium chloride (178.5mg, 4.2mmol) was added, the mixture was heated to 100 ℃ and reacted for 3 hours, 10mL of water and 10mL of dichloromethane were added, the mixture was stirred and the layers were separated, the aqueous layer was extracted with dichloromethane (10mL X3), the organic layers were combined, washed with saturated brine (50mL), and dried over anhydrous sodium sulfate. Filtration, spin-drying and reverse phase preparative purification of the residue gave example 31(22mg, white solid) after lyophilization. LCMS ESI (+) M/z 445.1(M +1).

Example 32(2171)

1- (3-fluoro-6, 11-dihydro-1, 2,11a triazabenzo [ cd, g ] azulen-6-yl) -5-hydroxy-3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione

The specific reaction equation is as follows:

step A: 1-bromo-2, 4-difluoro-3-nitrobenzene (Compound 32.1)

2, 6-difluoro-nitrobenzene (2g, 12.57mmol) was weighed out and dissolved in 16mL of sulfuric acid, and N-bromosuccinimide (3.41g, 15.09mmol) was added thereto and reacted at 80 ℃ under nitrogen for 3 hours. The reaction solution was cooled to 0 ℃ and poured into ice water, extracted 3 times with ethyl acetate (60mL), and the combined organic phases were washed 3 times with sodium bicarbonate solution (30mL), 2 times with saturated brine (30mL), and dried over anhydrous sodium sulfate. Filtration and concentration under reduced pressure, and purification of the residue by column chromatography (petroleum ether) gave 2.5g of the product as a yellow oil, yield: 83 percent.

And B: n-benzyl-6-bromo-3-fluoro-2-nitroaniline (Compound 32.2)

Compound 32.1(2.5g, 10.50mmol) was dissolved in 40mL of acetonitrile, benzylamine (1.13g, 10.50mmol) and potassium carbonate (2.9g, 21.01mmol) were added, and the mixture was stirred at 80 ℃ for 3 hours. Water (30mL) was added to the reaction mixture, which was extracted 2 times with ethyl acetate (40mL), and the combined organic phases were washed 2 times with saturated brine (20mL) and dried over anhydrous sodium sulfate. Filtration and concentration under reduced pressure, and column chromatography (petroleum ether: ethyl acetate: 20:1) of the residue afforded the product (2.5g, yellow oil, yield: 73%). LCMS ESI (+) M/z 325.0,327.0(M +1).

And C: n-benzyl-6-bromo-3-fluorobenzene-1, 2-diamine (Compound 32.3)

Compound 32.2(2.5g, 7.69mmol) was dissolved in 50mL of acetic acid, and iron powder (4.3g, 76.9mmol) was added in portions under nitrogen, stirred at 25 ℃ for 0.5 hour, then at 75 ℃ for 1 hour. The reaction solution was concentrated under reduced pressure, ethyl acetate (100mL) and water (40mL) were added to the residue, which was filtered through Celite, and the pH of the filtrate was adjusted to 8 with sodium bicarbonate. The organic phase was washed 2 times with saturated brine (20m l), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (petroleum ether/ethyl acetate 20/1) to give 32.3(1.8g, yellow oil, yield: 79%). LCMS ESI (+) M/z 295.0,297.0(M +1).

Step D: 1-benzyl-7-bromo-4-fluoro-1H-benzo [ d ] [1,2,3] triazole (Compound 32.4)

Compound 32.3(1.8g, 6.10mmol) was dissolved in 20ml of acetic acid, sodium nitrite (463mg, 6.71mmol) was added at room temperature, and the reaction mixture was stirred at room temperature for 1 hour. The reaction was cooled to 0 ℃ and the pH was adjusted to 9 with sodium bicarbonate solution. Extraction was performed 2 times with ethyl acetate (100mL), the combined organic phases were washed 2 times with saturated brine (20mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (petroleum ether/ethyl acetate: 10/1) to give 32.4(940mg, yellow solid, yield: 50%). LCMS ESI (+) M/z 306.0,308.0(M +1).

Step E: 1-benzyl-4-fluoro-1H-benzo [ d ] [1,2,3] triazole-7-carboxylic acid methyl ester (Compound 32.5)

Compound 32.4(400mg, 1.31mmol) was dissolved in 7.5ml of methanol, 2.5ml of N, N-dimethylformamide and 2.5ml of triethylamine, and 1,1' -bisdiphenylphosphinoferrocene palladium dichloride (200mg, 0.25mmol) and dicobalt octacarbonyl (400mg, 1.20mmol) were added. The reaction solution was stirred at 80 ℃ under nitrogen for 12 hours. The reaction mixture was filtered through celite, the filtrate was concentrated under reduced pressure, ethyl acetate (60ml) was added, washing was performed 2 times with water (20ml), washing was performed 3 times with saturated brine (20ml), drying was performed over anhydrous sodium sulfate, filtration was performed, concentration was performed under reduced pressure, and the residue was purified by column chromatography (petroleum ether/ethyl acetate: 10/1) to give 32.5(150mg, yield: 40%). LCMS ESI (+) M/z 286.1(M +1).

Step F: 1-benzyl-4-fluoro-1H-benzo [ d ] [1,2,3] triazole-7-carboxylic acid (Compound 32.6)

Compound 32.5(150mg, 0.53mmol) was dissolved in 10ml of tetrahydrofuran, and 3ml of an aqueous solution of lithium hydroxide monohydrate (88mg, 2.12mmol) was added. The reaction mixture was stirred at 45 ℃ for 1 hour. The reaction mixture was concentrated under reduced pressure to remove tetrahydrofuran, 15ml of water was added, saturated citric acid was added to adjust pH to 5, and extraction was performed 2 times with ethyl acetate (40 ml). The combined organic phases were washed with saturated brine (20ml), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the product 32.6(108mg, yellow solid, yield: 76%). LCMS ESI (+) M/z 272.1(M +1).

Step G: 3-fluoro 1,2,11a triazabenzo [ cd, g ] azulene-6- (11H) -one (Compound 32.7)

Compound 32.6(100mg, 0.37mmol) was added to 5ml of polyphosphoric acid and stirred for 2 hours at 120 degrees under nitrogen. To the reaction mixture were added water (30ml) and ethyl acetate (30 ml). The organic phase was adjusted to pH 9 with saturated sodium bicarbonate solution, washed with saturated brine (20ml), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (petroleum ether/ethyl acetate 3/1) to give 32.7(50mg, yellow solid, yield: 54%). LCMS ESI (+) M/z:254.1(M +1)

Step H: 3-fluoro-6, 11-dihydro-1, 2,11a triazabenzo [ cd, g ] azulen-6-ol (Compound 32.8)

Compound 32.7(50mg, 0.20mmol) was dissolved in 8ml tetrahydrofuran and sodium borohydride (30mg, 0.60mmol) was added at 0 ℃. The reaction solution was stirred at 0 ℃ for 0.5 hour. To the reaction mixture was added a saturated ammonium chloride solution (5ml), which was extracted with ethyl acetate (40ml), and the organic phase was washed with a saturated brine (15ml), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure, and the residue was purified by column chromatography (petroleum ether/ethyl acetate: 1/1) to give 32.8(33mg, white solid, yield: 65%). LCMSISI (+) M/z 256.1(M +1).

Step I: 5- (benzyloxy) -1- (3-fluoro-6, 11-dihydro-1, 2,11a triaza-benzo [ cd, g ] azulen-6-yl) -3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione (Compound 32.9)

Compound intermediate A (32mg, 0.13mmol) and compound 32.9(33mg, 0.15mmol) were dissolved in 3ml of 1-propylphosphoric anhydride/N, N-dimethylformamide 50%. The reaction solution was stirred at 100 ℃ under nitrogen for 1 hour. Water (10ml) was added to the reaction solution, extracted 2 times with ethyl acetate (30ml), the combined organic phases were washed with saturated brine (15ml), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (petroleum ether/ethyl acetate: 0/1) to give 32.9(20mg, yellow solid, yield: 30%). LCMS ESI (+) M/z:536.2(M +1).

Step J: 1- (3-fluoro-6, 11-dihydro-1, 2,11a triazabenzo [ cd, g ] azulen-6-yl) -5-hydroxy-3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione (Compound 32)

Compound 32.9(20mg, 0.04mmol) is dissolved in 4ml of N, N-dimethylformamide and lithium chloride (80mg, 1.87mmol) is added. The reaction solution was stirred at 100 ℃ under nitrogen for 6 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by reverse phase preparation to give product 32(9mg, pale yellow solid, yield: 54%). LCMS ESI (+) M/z:446.1(M +1).

Example 33(2172)

1- (3-fluoro-6, 11-dihydrobenzo [5,6] azepino [3,2,1-hi ] indazol-6-yl) -5-hydroxy-3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione

Step A: (E) -N- (3-fluoro-2-methylphenyl) -2- (hydroxyimino) acetamide (Compound A-2) (Compound 33.1)

Weighed anhydrous sodium sulfate (22.7g, 160mmol) was added to a single-neck flask containing 65mL of water, chloral hydrate (2.9g, 17.6mmol) was added to the mixture, and a solution of 3-fluoro-2-methylaniline (2g, 16mmol) in 1mol/L hydrochloric acid (19.2mL, 19.2mmol) was added dropwise to the reaction flask (at this time, the reaction solution became cloudy). Finally, hydroxylamine hydrochloride (4.45g, 64mmol) was added to the reaction flask, and the reaction solution was stirred for reaction for 3 hours under an oil bath at 80 ℃. The reaction was cooled to normal temperature and filtered to finally obtain a light brown solid crude product 33.1 (2.65g, yield: 84.5%). LCMS ESI (+) M/z 197.1(M +1).

And B: 6-fluoro-7-methylindoline-2, 3-dione (Compound 33.2)

Compound 33.1(1.65g, 8.42mmol) was dissolved in concentrated sulfuric acid (20mL) and the mixture was stirred in an oil bath at 80 deg.C for 4 hours. The reaction solution was poured into ice-water, followed by extraction with ethyl acetate (15mL), and the combined organic phases were dried over anhydrous sodium sulfate. Filtration, spin-drying and purification by column chromatography gave 33.2(950mg, yield: 63.3%). LCMS ESI (+) M/z:180.2(M +1).

And C: synthesis of 2-amino-4-fluoro-3-methylbenzoic acid (Compound 33.3)

Compound 33.2(950mg, 5.31mmol) was dissolved in 25mL of water, and potassium hydroxide (327mg, 5.84mmol) and potassium chloride (831mg, 11.15mmol) were added to the reaction solution under ice bath. Then, 30% hydrogen peroxide (902.7mg, 7.97mmol) was slowly dropped into the reaction flask (the dropping time required was more than 30 minutes), and finally the reaction was stirred at room temperature overnight. The reaction solution was acidified to pH 5 with glacial acetic acid, extracted with ethyl acetate (15mL), and the organic phases were combined, separated, washed with saturated brine, and dried over anhydrous sodium sulfate. Filtration and spin-drying gave 33.3(710mg, yield: 79.15%) as an orange solid. LCMSISI (+) M/z 170.2(M +1).

Step D: 2-amino-4-fluoro-3-methylbenzoic acid methyl ester (Compound 33.4)

Compound 33.3(710mg, 4.2mmol) was dissolved in methylene chloride/methanol (12mL, 5:1), trimethylsilyldiazomethane (3.15mL, 6.3mmol) was added dropwise to the reaction mixture while cooling on ice, and the reaction was stirred at room temperature for 1.5 hours. The reaction was quenched with water, extracted with dichloromethane (12mL), and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, spun dry, and purified to give 33.4 as a pale yellow liquid (690mg, yield: 89.8%). LCMS ESI (+) M/z:184.2(M +1).

Step E: 4-fluoro-1H-indazole-7-carboxylic acid methyl ester (33.5)

Compound 33.4(690g, 3.77mmol) was dissolved in glacial acetic acid (20mL), sodium nitrite (286mg, 4.15mmol) was dissolved in water (1.5mL) and added dropwise to the reaction flask, and the reaction was stirred at room temperature for 1.5 hours. The reaction mixture was concentrated, basified to pH 7 with a saturated sodium bicarbonate solution, extracted with ethyl acetate (15mL), the organic phases were separated and combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and spin-dried to give 33.5(490mg, yield: 67%) as a white solid. LCMS ESI (+) M/z 195.2(M +1).

Step F: 1-benzyl-4-fluoro-1H-indazole-7-carboxylic acid methyl ester (compound 33.6)

Compound 33.5(490mg, 2.53mmol) was dissolved in N, N-dimethylformamide (10mL) and cesium carbonate (1.24g, 3.8mmol) and benzyl bromide (520mg, 3.04mmol) were added to the reaction flask, followed by stirring the reaction in an 80 ℃ oil bath for 1 hour. The reaction was quenched with water, then extracted with ethyl acetate (12mL), and the organic phases were separated, combined, dried over anhydrous sodium sulfate, filtered, spun dry, and purified to give 33.6(430mg, yield: 59.97%) as a colorless liquid. LCMS ESI (+) M/z 285.3(M +1).

Step G: 1-benzyl-4-fluoro-1H-indazole-7-carboxylic acid (compound 33.7)

Compound 33.6(430mg, 1.51mmol) was dissolved in tetrahydrofuran/methanol (6mL, 5:1), 3mol/L sodium hydroxide solution was added to the reaction flask, and the reaction was stirred in a 60 ℃ oil bath for 1 hour. The reaction mixture was concentrated, water (10mL) was added, and then acidified to pH 3-4 with 2mol/L hydrochloric acid solution, filtered, and dried to give 33.7(330mg, yield: 80.72%) as a white solid. LCMS ESI (+) M/z 271.3(M +1).

Step H: 3-fluoro-benzo [5,6] azabicyclo [3,2,1-H i ] indazol-6 (11H) -one (Compound 33.8)

Compound 33.7(330mg, 1.22mmol) was dissolved in polyphosphoric acid (5mL) and the reaction was left to stir for 2 hours in a 120 ℃ oil bath. Ice water was added to the reaction flask to give a white solid, which was filtered and spun to give 33.8(250mg, yield: 81.17%) as a white solid. LCMS ESI (+) M/z:253.2(M +1).

Step I: synthesis of 3-fluoro-6, 11-dihydrobenzo [5,6] azepino [3,2,1-hi ] indazol-6-ol (Compound 33.9)

Compound 33.8(250mg, 1mmol) was dissolved in tetrahydrofuran/methanol (6mL, 5:1), sodium borohydride (38mg, 1mmol) was added to the reaction flask under ice bath, and the reaction was stirred at room temperature for 30 minutes. After quenching the reaction with saturated ammonium chloride solution, water (10mL) was added to dissolve the precipitated salt, followed by extraction with ethyl acetate (12mL), the organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, filtered, spun-dried, and purified to give 33.9(250mg, yield: 99.2%) as a pale yellow solid. LCMS ESI (+) M/z 255.3(M +1).

Step J: 5- (benzyloxy) -1- (3-fluoro-6, 11-dihydrobenzo [5,6] azepino [3,2, 1-xi ] indazol-6-yl) -3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione compound () compound 33.10)

Intermediate a (25mg, 0.084mmol) and compound 33.9(28mg, 0.11mmol) were dissolved in 3mL of 1-propylphosphoric anhydride (50% ln DMF), then the reaction was placed in a 100 ℃ oil bath and stirred for 1.5 h-confirmation of reaction completion by TLC (DCM: MeOH ═ 10:1) and LCMS water (12mL) was added to the reaction flask, followed by extraction with ethyl acetate (12mL), washing with saturated brine, drying over anhydrous sodium sulfate, filtration, and purification over silica gel column gave 33.10(30mg, yield: 66.96%) LCMS ESI (+) M/z:535.2(M +1).

Step K: synthesis of Compound 33

Compound 33.10(30mg, 0.056mmol) was dissolved in 5mL of N, N-dimethylformamide solution, then lithium chloride (117.6mg, 2.8mmol) was added and the reaction was stirred in a 100 ℃ oil bath for 4 hours. After completion of the reaction, water (15mL) was added to the reaction solution, which was then extracted with ethyl acetate (10mL), washed with saturated brine, and dried over anhydrous sodium sulfate. Filtration, concentration and purification gave example 33(17mg, yield: 68.16%) as a pale yellow solid. LCMS ESI (+) M/z 445.2(M +1).

Example 34(2187)

1' - (3-fluoro-6, 11-dihydrobenzo [5,6] azepino [3,2,1-hi ] indazol-6-yl) -5' -hydroxy-1 ', 2' -dihydrospiro [ cyclobutane-1, 3' -pyrido [1,2-b ] pyridazine ] -4', 6' -dione

Example 34 was obtained by a two-step synthesis, using intermediate B instead of intermediate A and compound 33.9, according to the synthesis procedure of reference example 33

LCMS ESI(+)m/z:457.2(M+1)。

Example 35(2155)

1- (2-chloro-4, 10-dihydrobenzo [ b ] thieno [3,2-e ] thiepin-4-yl) -5-hydroxy-3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione

Step A (5-chlorothien-2-yl) methanol (Compound 35.1)

The compound 5-chlorothiophene-2-carbaldehyde (2g,13.64mmol) was dissolved in 15mL of a mixed solvent of tetrahydrofuran and 2mL of methanol, and a solid of sodium borohydride (773mg,20mmol) was added in portions under ice bath, followed by stirring at room temperature for 10 minutes. After the reaction is completed, adding saturated ammonium chloride aqueous solution for quenching, extracting with 30mL ethyl acetate for three times respectively, combining organic phases, washing with saturated saline solution, and drying with anhydrous sodium sulfate. Filtration and spin-drying gave compound 35.1(1.91g, yield: 94.2%). LCMS ESI (+) M/z:149.1(M +1).

And B:2- (bromomethyl) -5-chlorothiophene (Compound 35.2)

Compound 35.1(1.91g, 12.85mmol) was dissolved in 30mL of dichloromethane, phosphorus tribromide (3.5g,12.93mmol) was slowly added dropwise in ice bath, and stirred for 10 minutes in ice bath, followed by addition of water to quench the reaction. Extraction was performed with 30mL of dichloromethane, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and then spin-dried to obtain compound 35.2(2.26g, yield: 83.15%) as a yellow oil. LCMS ESI (+) M/z:210.2(M +1).

And C: methyl 2- (((5-chlorothien-2-yl) methyl) thio) benzoate (Compound 35.3)

Compound 35.2(1g,3.35mmol) and methyl 2-mercaptobenzoate (960mg,6.15mmol) were dissolved in 5mL of N, N-dimethylformamide, potassium carbonate (990mg, 7.17mmol) was added under ice bath, and stirred for 30 minutes. The reaction mixture was diluted with 30mL of water, extracted three times with 20mL of ethyl acetate, and the organic phases were combined, washed with saturated brine, and dried over anhydrous sodium sulfate. Filtration and spin-drying of the solvent gave compound 35.3(1.46g) as a crude product. LCMS ESI (+) M/z 299.1(M +1).

Step D:2- (((5-chlorothien-2-yl) methyl) thio) benzoic acid (Compound 35.4)

Compound 35.3(1.46g, crude) was dissolved in 15mL tetrahydrofuran and 2mL methanol, then sodium hydroxide solution (4M, 5mL) was added and stirred at room temperature overnight. After completion of the reaction, the reaction mixture was spin-dried, water (30mL) was added to dissolve the solid, the mixture was acidified with hydrochloric acid (pH 3), extracted 3 times with ethyl acetate (20mL), and washed 1 time with saturated brine (30 mL). After purification by drying, the product 35.4(1.1g, two-step yield: 85%) was obtained. LCMS ESI (+) M/z 285.1(M +1).

Step E: 2-chlorobenzo [ b ] thieno [3,2-e ] thiaheptacyclo-4 (10H) -one (compound 35.5)

Compound 35.4(100mg,0.35mmol) was dissolved in 2g of polyphosphoric acid and 3mL of sulfolane, heated to 100 ℃ and stirred for 10 minutes. Water (30mL) was added to the reaction mixture, which was extracted three times with ethyl acetate (15mL) and washed with sodium bicarbonate solution (20 mL). Drying the anhydrous sodium sulfate. Filtration and spin-drying gave compound 35.5(68mg, yield: 72.8%). LCMS ESI (+) M/z 267.1(M +1).

Step F: 2-chloro-4, 10-dihydrobenzo [ b ] thieno [3,2-e ] thiepin-4-ol (Compound 35.6)

Compound 35.5(68mg,0.26mmol) was dissolved in 3mL of tetrahydrofuran and 1mL of methanol under ice-bath conditions, followed by addition of sodium borohydride (15mg, 0.40mmol) and stirring was continued for 0.5 h while maintaining the temperature. Quenched by addition of saturated ammonium chloride solution, extracted 2 times with ethyl acetate (15mL), washed 1 time with saturated brine (20mL), and the organic phase dried over anhydrous sodium sulfate. Filtration and concentration gave 35.6 (68mg, yield: 99.2%) as the crude title product. LCMS ESI (+) M/z 269.1(M +1).

Step G: 5- (benzyloxy) -1- (2-chloro-4, 10-dihydrobenzo [ b ] thieno [3,2-e ] thiepin-4-yl) -3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione (Compound 35.7)

Intermediate A (90mg, 0.30mmol) was dissolved in 3mL of N, N-dimethylformamide, then compound 106.6 (68mg,0.25mmol) and a solution of 1-propylphosphoric anhydride in N, N-dimethylformamide (50%, 3mL) were added to the reaction flask and the reaction was stirred at 100 ℃ for 0.5 h. The reaction was quenched with saturated sodium bicarbonate solution, the aqueous phase was extracted with ethyl acetate (15mLX2), and the organic phases were combined and dried over anhydrous sodium sulfate. Filtration and purification by rotary drying gave 35.7(61mg, yield: 43.9%). LCMS ESI (+) M/z 549.2(M +1).

Step H: 1- (2-chloro-4, 10-dihydrobenzo [ b ] thieno [3,2-e ] thiepin-4-yl) -5-hydroxy-3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione (example 35)

Compound 35.7(61mg,0.11mmol) was dissolved in 4mL of N, N-dimethylformamide, lithium chloride (100mg, 2.38mmol) was added, the reaction was heated to 100 ℃ for 1 hour, 10mL of water and 10mL of ethyl acetate were added, the layers were separated by stirring, the aqueous layer was extracted with ethyl acetate (10mL X3), the organic layers were combined, washed with saturated brine (50mL), and dried over anhydrous sodium sulfate. Filtration, spin-drying, purification by reverse phase preparative lyophilization gave product example 35(2.4mg, yield: 4.5%). LCMS ESI (+) M/z 459.1(M +1).

Example 3692154)

1- (4, 10-dihydrobenzo [ b ] thieno [3,2-e ] thiepin-4-yl) -5-hydroxy-3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2b ] pyridazine-4, 6-dione

The specific reaction equation is as follows:

step A: 2-Formylthiophene-3-carboxylic acid (Compound 36.1)

Thiophene-3-carboxylic acid (3.48g,27.16mmol), a raw material, was dissolved in 50mL of tetrahydrofuran, 10mL of tetramethylethylenediamine was added under a nitrogen atmosphere at-78 ℃ and stirred for 10 minutes, then N-butyllithium (2.5M,27mL) was added dropwise and stirred for one hour, and finally 4.6mL of N, N-dimethylformamide was added dropwise and stirred for 30 minutes. The reaction mixture was quenched by addition of dilute hydrochloric acid solution, extracted three times with 30mL of ethyl acetate, respectively, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and dried by rotary drying to give 36.1(4.17g, yield 98.3%). LCMS ESI (+) M/z:157.1(M +1).

And B: 2-Formylthiophene-3-carboxylic acid ethyl ester (Compound 36.2)

Compound 36.1(2.1g, 13.45mmmol) was dissolved in 30mL of N, N-dimethylformamide, potassium carbonate (3.54g,25.65mmol) and iodoethane (2.2g,14.10mmol) were added at room temperature, and the mixture was heated to 50 ℃ and stirred for 3 hours. Then, it was diluted with water, extracted three times with 30mL of ethyl acetate, respectively, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and spin-dried to obtain 36.2(2.17g, yield: 87.6%) as a product. LCMS ESI (+) M/z 185.1(M +1).

Step C2- (hydroxymethyl) thiophene-3-carboxylic acid ethyl ester (Compound 36.3)

Compound 107.2(1g, 5.43mmol) was dissolved in a mixed solvent of 15mL of tetrahydrofuran and 2mL of methanol, and sodium borohydride (310mg,8.16mmol) was added as a solid in portions under ice bath, followed by stirring at room temperature for 10 minutes. After the reaction was completed, saturated aqueous ammonium chloride was added thereto and quenched, and extracted with 30mL of ethyl acetate three times, respectively, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and spin-dried to obtain 36.3(1.01g, yield: 99.8%). LCMS ESI (+) M/z 187.1(M +1).

Step D:2- (bromomethyl) thiophene-3-carboxylic acid ethyl ester (Compound 36.4)

Compound 36.3(1.01g,5.42mmol) was dissolved in 30mL of dichloromethane, phosphorus tribromide (484mg, 1.79mmol) was slowly added dropwise in ice bath and stirred for 10 minutes in ice bath, and then water was added to quench the reaction. After extraction with 30mL of dichloromethane, the organic phases were combined, washed with brine, dried over anhydrous sodium sulfate, and then dried by spin-drying, compound 36.4(930mg, yield: 68.9%) was obtained. LCMS ESI (+) M/z:249.2(M +1).

Step E: 2- ((phenylthio) methyl) thiophene-3-carboxylic acid ethyl ester (Compound 107.5)

Compound 36.4(930mg,3.73mmol) and thiophenol (453mg,4.11mmol) were dissolved in 5mL of N, N-dimethylformamide, and potassium carbonate (776mg, 5.62mmol) was added in an ice bath and stirred for 30 minutes. The reaction mixture was diluted with 30mL of water, extracted three times with 20mL of ethyl acetate, and the organic phases were combined, washed with saturated brine, and dried over anhydrous sodium sulfate. Filtration and spin-drying of the solvent gave compound 36.5(900mg, yield: 86.7%). LCMS ESI (+) M/z 279.1(M +1).

Step F: 2- ((phenylthio) methyl) thiophene-3-carboxylic acid (Compound 36.6)

Compound 36.5(900mg, 3.23mmol) was dissolved in 10mL of tetrahydrofuran and 1mL of methanol, followed by addition of 4mL of sodium hydroxide solution (4M) and stirring at room temperature overnight. After completion of the reaction, the reaction mixture was spin-dried, water (30mL) was added to dissolve the solid, the mixture was acidified with hydrochloric acid (pH 3), extracted 3 times with ethyl acetate (20mL), and washed with saturated brine (30 mL). After purification by drying, the product was obtained as 36.6(790mg, yield: 97.7%). LCMS ESI (+) M/z:251.1(M +1).

Step G: benzo [ b ] thieno [3,2-E ] thiaheptacyclo-4 (10H) -one (compound 36.7)

Compound 36.6(790mg, 3.16mmol) was dissolved in polyphosphoric acid (2g) and 3mL of sulfolane and stirred for 10 minutes at 120 ℃. Water (30mL) was added to the reaction mixture, which was extracted three times with 15mL of ethyl acetate. The organic phases were combined and washed with sodium bicarbonate solution (20 mL). The organic phase was dried over anhydrous sodium sulfate. Filtration and spin-drying gave 36.7(210mg, yield: 28.6%). LCMS ESI (+) M/z:233.1(M +1).

Step H: 4, 10-dihydrobenzo [ b ] thieno [3,2-e ] thiepin-4-ol (Compound 36.8)

Under ice-bath conditions, compound 107.7(50mg,0.20mmol) was dissolved in 3mL of tetrahydrofuran and 1mL of methanol, followed by addition of sodium borohydride (15mg, 0.395mmol) and stirring was continued for 0.5 h while maintaining the temperature. The mixture was quenched by addition of saturated ammonium chloride solution, extracted 2 times with ethyl acetate (15mL) and washed 1 time with saturated brine (20 mL). Dried over anhydrous sodium sulfate. Filtration and concentration gave 36.8(50mg, yield: 99.5%) as the desired product. LCMS ESI (+) M/z:235.1(M +1).

Step I: 5- (benzyloxy) -1- (4, 10-dihydrobenzo [ b ] thieno [3,2-e ] thiepin-4-yl) -3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,22-b ] pyridazine-4, 6-dione (Compound 36.9)

Intermediate A (50mg,0.171mmol) was dissolved in 3mL of N, N-dimethylamide, then compound 36.8(51mg, 0.198mmol) and a solution of 1-propylphosphoric anhydride in N, N-dimethylamide (50%, 3mL) were added to the reaction flask and the reaction stirred at 100 ℃ for 0.5 h. The reaction was quenched with saturated sodium bicarbonate solution, and the aqueous phase was extracted with ethyl acetate (15mL) 2 times, dried over anhydrous sodium sulfate, filtered, spun-dried, and purified to give the product 36.9(10mg, yield: 9.7%). LCMS ESI (+) M/z 515.2(M +1).

Step J: 1- (4, 10-Dihydrobenzo [ b ] thieno [3,2-e ] thiepin-4-yl) -5-hydroxy-3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2b ] pyridazine-4, 6-dione (example 36)

Compound 36.9(60mg,0.116mmol) was dissolved in 4mL of N, N-dimethylformamide, lithium chloride (60mg,0.353mmol) was added, the mixture was heated to 100 ℃ and reacted for 1 hour, 10mL of water and 10mL of ethyl acetate were added, the mixture was stirred and the layers were separated, the aqueous layer was extracted with ethyl acetate (10mL X3), the organic layers were combined, washed with saturated brine (50mL), and dried over anhydrous sodium sulfate. Filtration, spin-drying, purification by reverse phase preparation and lyophilization gave the product, example 36(20mg, yield: 40.6%). LCMS ESI (+) M/z:425.1(M +1).

Example 37(2168)

1- (6, 11-dihydrobenzo [5,6] azepino [3,2,1-H i ] indazol-6-yl) -5-hydroxy-3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione

Step A: methyl 1H-indazole-7-carboxylic acid methyl ester (Compound 37.1)

Methyl 2-amino-3-methylbenzoate (2g, 12.18mmol) was dissolved in 20mL of acetic acid solution, 5mL of water was added, sodium nitrite (1.25g, 18.18mmol) was slowly added, and stirring was carried out at room temperature for 2 hours. The reaction was quenched by addition of saturated sodium bicarbonate solution, extracted three times with 100ml ethyl acetate, the organic phases combined, spun dried and purified to give 37.1(460mg, 22%). LCMS ESI (+) m/z:177.1(M +1)

And B: 1-benzyl-1H-indazole-7-carboxylic acid methyl ester (Compound 37.2)

Compound 37.1(460mg, 2.6mmol) was dissolved in 10mL of N, N-dimethylformamide, benzyl bromide (670mg, 3.9mmol), cesium carbonate (1.3g, 3.9mmol) and nitrogen were added, and the reaction was stirred at 80 ℃ for 2 hours. The reaction was quenched by addition of aqueous solution, extracted three times with 100ml ethyl acetate, the organic phases combined, spun dried and purified to give 37.2(360mg, 52%). LCMS ESI (+) m/z:267.1(M +1)

And C: 1-benzyl-1H-indazole-7-carboxylic acid (Compound 37.3)

Compound 37.2(360mg, 1.35mmol) was dissolved in a solution of methanol (10mL) and water (10mL), lithium hydroxide (155mg, 3.7mmol) was added, and the reaction mixture was stirred at room temperature overnight. The reaction solution was adjusted to pH 5 with HCl (2M), a solid precipitated, filtered and the solid dried to give 37.3(300mg, 88%). LCMS ESI (+) m/z: 253.1(M +1)

Step D: benzo [5,6] azabicyclo [3,2, 1-xi ] indazol-6 (11H) -one (compound 37.4)

Compound 37.3(300mg, 1.2mmol) was charged in a 100ml round-bottom flask, PPA (15ml) was added, and the reaction was stirred at 120 ℃ for 3 hours. Cooled to room temperature, quenched with 50ml of water, extracted three times with 100ml of ethyl acetate, the organic phases combined, spun dry and purified to give 37.4(160mg, 57%). LCMS ESI (+) m/z:235.1(M +1).

Step E: 6, 11-Dihydrobenzo [5,6] azepino [3,2, 1-xi ] indazol-6-ol (Compound 37.5)

Compound 37.4(160mg, 0.68mmol) was dissolved in 10mL tetrahydrofuran and 2mL methanol solution, the reaction was cooled to 0 ℃, sodium borohydride (32mg, 0.82mmol) was added in portions, stirred at room temperature for 0.5 h, quenched by addition of saturated aqueous ammonium chloride, extracted three times with 100m l ethyl acetate, the organic phases were combined, spun dry and purified to give product 37.5(120mg, 74%). LCMS ESI (+) m/z: 237.1(M +1).

Step A:5- (benzyloxy) -1- (6, 11-dihydrobenzo [5,6] azepino [3,2, 1-xi ] indazol-6-yl) -3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione (Compound 37.6)

Compound 37.5(60mg, 0.25mmol) was dissolved in 5mL of N, N-dimethylformamide solution, and intermediate A (63mg, 0.2mmol) and 5mL of 1-propylphosphoric anhydride 50% N, N-dimethylformamide solution were added to stir the reaction solution at 100 ℃ for 2 hours. The reaction was quenched by addition of water (20ml), extracted three times with 100ml ethyl acetate, the organic phases combined, spun dry and purified to give 37.6(70mg, 64%). LCMS ESI (+) m/z: 517.2(M +1)

And B:1- (6, 11-Dihydrobenzo [5,6] azepino [3,2,1-H i ] indazol-6-yl) -5-hydroxy-3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione (Compound 37)

Compound 37.6(70mg, 0.14mmol) was dissolved in N, N-dimethylformamide (10ml), lithium chloride (88mg, 2.1mmol) was added, the reaction solution was stirred at 100 ℃ for 4 hours, water (20ml) was added to quench the reaction, extraction was carried out three times with 100ml ethyl acetate, and the organic phases were combined, spun dry and purified to give example 37(10mg, 17%). LCMS ESI (+) m/z: 427.2(M +1)

Example 38(2186)

1' - (6, 11-dihydrobenzo [5,6] azepino [3,2,1-hi ] indazol-6-yl) -5' -hydroxy-1 ', 2' -dihydrospiro [ cyclobutane-1, 3' -pyrido [1,2-b ] pyridazine ] -4', 6' -dione

Reference example 37 reaction of intermediate B instead of intermediate A with Compound 37.5 gave Compound 38 via a two-step synthesis

LCMS ESI(+)m/z：439.2(M+1).

Example 39(2169)

1- (6, 11-dihydro-2, 11 a-dibenzo [ cd, g ] azulen-6-yl) -5-hydroxy-3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2b ] pyridazine-4, 6-dione

Step A:2- (benzylamino) -3-nitrobenzoic acid methyl ester (39.1)

The compound methyl 2-fluoro-3-nitrobenzoate (2g, 10.05mmol) was dissolved in 20mL of N, N-dimethylformamide, benzylamine (1.3g, 12.06mmol), potassium carbonate (2.0g, 15.07mmol) and nitrogen were added, and the reaction was stirred at 80 ℃ for 2 hours. After cooling to room temperature, the reaction was quenched by addition of water (80ml), extracted three times with 100ml ethyl acetate, the organic phases were combined, spun dry and purified to give product 39.1(2.2g, 77%). LCMS ESI (+) m/z:287.1(M +1)

And B: 3-amino-2- (benzylamino) benzoic acid methyl ester (39.2)

Compound 39.1(2.2g, 7.7mmol) was dissolved in a methanol (20ml) solution, palladium on carbon (200mg, 0.8mmol) was added, and the mixture was stirred at room temperature for 5 hours under hydrogen pressure. The reaction was filtered, the solvent was spun off, and the product was purified to give 39.2(670mg, 34%). LCMS ESI (+) m/z:257.1(M +1)

And C: 1-benzyl-1H-benzo [ d ] imidazole-7-carboxylic acid methyl ester (Compound 39.3)

Compound 39.2(670mg, 2.6mmol) was dissolved in formic acid (10ml), the reaction was heated to 90 ℃ and stirred for 1 hour, the solvent was dried by spinning, water (40ml) was added, extraction was carried out three times with 100ml ethyl acetate, the organic phases were combined, spun and purified to give product 39.3(700 g, 100%). LCMS ESI (+) m/z:267.1(M +1)

Step D: 1-benzyl-1H-benzo [ d ] imidazole-7-carboxylic acid (Compound 39.4)

Compound 39.3(700mg, 2.6mmol) was dissolved in a solution of methanol (10ml) and water (10ml), lithium hydroxide (310mg, 7.2mmol) was added, and the reaction solution was stirred at room temperature overnight. The reaction solution was adjusted to pH 5 with HCl (2M), a solid precipitated, filtered and the solid dried to give 39.4(590mg, 43%). LCMS ESI (+) m/z: 253.1(M +1)

Step E: 2,11 a-dibenzo [ CD, G ] azulene-6- (11H) -one (Compound 39.5)

Compound 39.4(590mg, 2.3mmol) was charged into a 100mL round-bottom flask, PPA (20mL) was added, and the reaction was stirred at 120 ℃ for 3 hours. Cooled to room temperature, quenched with 50ml of water, extracted three times with 100ml of ethyl acetate, the organic phases combined, spun dry and purified to give 39.5(430mg, 79%). LCMS ESI (+) m/z:235.1(M +1)

Step F: 6, 11-dihydro-2, 11 a-dibenzo [ cd, g ] azulen-6-ol (Compound 39.6)

Compound 39.5(430, 1.84mmol) was dissolved in tetrahydrofuran (10ml), methanol (2ml) solution, the reaction was cooled to 0 ℃, sodium borohydride (84mg, 2.2mmol) was added in portions, stirred at room temperature for 0.5 h, quenched by addition of saturated aqueous ammonium chloride solution, extracted three times with 100ml ethyl acetate, the organic phases were combined, spun dry and purified to give product 39.6(400mg, 92%). LCMS ESI (+) m/z: 237.1(M +1)

Step G: 5- (benzyloxy) -1- (6, 11-dihydro-2, 11 a-dibenzo [ cd, g ] azulen-6-yl) -3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,22-b ] pyridazine-4, 6-dione (Compound 39.7)

Compound 39.6(83mg, 0.35mmol) was dissolved in 5mL of N, N-dimethylformamide, and intermediate A (70mg, 0.225mmol) was added followed by 5mL of 1-propylglycerophosphate (50% in DMF), and the reaction was stirred at 100 ℃ for 2 hours. The reaction was quenched by addition of water (20mL), extracted three times with 100mL ethyl acetate, the organic phases combined, spun dry, and purified to give product 39.7(100mg, 82.5%). LCMS ESI (+) m/z: 517.2(M +1).

Step H: 1- (6, 11-dihydro-2, 11 a-dibenzo [ cd, g ] azulen-6-yl) -5-hydroxy-3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2b ] pyridazine-4, 6-dione (Compound 39)

Compound 39.7(100mg, 0.19mmol) was dissolved in 10mL of N, N-dimethylformamide, lithium chloride (147mg, 3.8mmol) was added, the reaction mixture was stirred at 100 ℃ for 4 hours, water (20mL) was added to quench the reaction, extraction was carried out three times with 100mL of ethyl acetate, the organic phases were combined, spun dry and purified to give example 39(15mg, 18%). LCMS ESI (+) m/z: 427.2(M +1).

Example 40(2178)

1' - (7, 12-dihydro-6H-4, 5,5a triazabenzo [5,6] cycloocta [1,2,3- ] inden-12-yl) -5' -hydroxy-1 ', 2' dihydrospiro [ cyclobutane-1, 3' -pyrido [1,2-b ] pyridazine ] -4', 6' -dione

Step A:2- (benzylamino) -3-nitrobenzoic acid methyl ester (Compound 40.1)

1-fluoro-2-nitro-benzoic acid methyl ester (1g, 5.0mmol) was weighed out and dissolved in 10mL of N, N-dimethylformamide, potassium carbonate (1.0g, 7.5mmol) and phenethylamine (730mg, 6.0mmol) were added and stirred overnight at 80 ℃ under nitrogen. To the reaction mixture was added ethyl acetate (60mL), washed 2 times with saturated brine (20mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (petroleum ether: ethyl acetate 5:1) to give the product 40.1(1.2mg, 80%). LCMS ESI (+) M/z:301.1(M +1).

And B: 3-amino-2- (phenethylamino) benzoic acid methyl ester (Compound 40.2)

Compound 40.1(1.2g, 4.0mmol) was dissolved in methanol (15m l), palladium on carbon (120mg, 0.1mmol) was added, and the mixture was stirred at room temperature for 5 hours under hydrogen pressure. The reaction was filtered, the solvent was spun off, and the product was purified to give 40.2(600mg, 56%). LCMS ESI (+) m/z: 271.1(M +1)

And C: 1-phenethyl-1H-benzo [ d ] [1,2,3] triazole-7-carboxylic acid methyl ester (Compound 40.3)

Compound 40.2(600mg, 2.22mmol) was dissolved in 20mL of acetic acid, sodium nitrite (184mg, 2.66mmol) was added at room temperature, and the reaction mixture was stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure, and ethyl acetate (100mL) was added to the residue to adjust the pH to 8 with sodium bicarbonate solution. The organic phase was washed 2 times with saturated brine (20mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (petroleum ether: ethyl acetate 3:1) to give the product 40.3(550mg, 88%). LCMS ESI (+) M/z 282.1(M +1).

Step D: 1-phenethyl-1H-benzo [ d ] [1,2,3] triazole-7-carboxylic acid (Compound 40.4)

Compound 40.3(550mg, 1.96mmol) was dissolved in 10mL of methanol, and 3mL of an aqueous solution containing lithium hydroxide monohydrate (200mg, 7.8mmol) was added. The reaction solution was stirred at room temperature for 3 hours. The reaction mixture was concentrated under reduced pressure to remove methanol, 15mL of water was added, a saturated citric acid solution was added to adjust the pH to 5, and a solid was precipitated, filtered, and dried to obtain 40.4(430mg, 82%) as a product. LCMS ESI (+) m/z:268.1(M +1)

Step E: 6H-4,5,5a triaza-benzo [5,6] cycloocta [1,2,3-cd ] indene-12 (7H) -one (Compound 40.5)

Compound 40.4(200mg, 0.75mmol) was added to 10mL of polyphosphoric acid and heated to 120 ℃ under nitrogen and stirred for 4 hours. After cooling to room temperature, water (30mL) and ethyl acetate (30mL) were added to the reaction mixture. The organic phase was adjusted to pH 9 with saturated sodium bicarbonate solution, washed with saturated brine (20mL) and dried over anhydrous sodium sulfate. Filtration and purification by concentration under reduced pressure gave 40.5(170mg, 91%). LCMS ESI (+) M/z:250.1(M +1)

Step F: 6H-4,5,5a triaza-benzo [5,6] cycloocta [1,2,3-CD ] indene-12 (7H) -ol (Compound 40.6)

Compound 40.5(170mg, 0.68mmol) was dissolved in 10mL tetrahydrofuran and sodium borohydride (31mg, 0.89mmol) was added at 0 deg.C. The reaction mixture was warmed to room temperature and stirred for 0.5 hour. The reaction mixture was quenched by addition of saturated ammonium chloride solution (5mL), extracted with ethyl acetate (40mL), and the organic phase was washed with saturated brine (15mL) and dried over anhydrous sodium sulfate. Filtration and concentration under reduced pressure, and column chromatography (petroleum ether: ethyl acetate: 1) of the residue yielded 40.6(150mg, yield: 88%). LCMS ESI (+) M/z 252.1(M +1).

Step G: 5' - (benzyloxy) -1' - (7, 12-dihydro-6H-4, 5,5a triaza-benzo [5,6] cycloocta [1,2,3-cd ] indene-12-yl) -1', 2' -dihydrospiro [ cyclobutane-1, 3' -pyrido [1,2-b ] pyridazine ] -4', 6' -dione (Compound 40.7)

Compound 40.6(70mg, 0.28mmol) and intermediate B (60mg, 0.18mmol) were dissolved in 5mL of 1-propylphosphoric anhydride (50% in DMF). The reaction solution was stirred at 100 ℃ under nitrogen for 1 hour. Water (10mL) was added to the reaction mixture, which was extracted 2 times with ethyl acetate (30mL), and the combined organic phases were washed with saturated brine (15mL) and dried over anhydrous sodium sulfate. Filtration and concentration under reduced pressure, and the residue was purified by column chromatography (petroleum ether: ethyl acetate: 10:1) to give the product 40.7(50mg, yield: 33%). LCMS ESI (+) M/z 544.2(M +1).

Step H: 1' - (7, 12-dihydro-6H-4, 5,5a triaza-benzo [5,6] cycloocta [1,2,3-cd ] inden-12-yl) -5' -hydroxy-1 ', 2' dihydrospiro [ cyclobutane-1, 3' -pyrido [1,2-b ] pyridazine ] -4', 6' -dione (Compound 40)

Compound 40.7(50mg, 0.09mmol) was dissolved in N, N-dimethylformamide (10m l), and lithium chloride (120mg, 2.8mmol) was added. The reaction solution was stirred at 100 ℃ under nitrogen for 5 hours. The reaction was concentrated under reduced pressure and the residue was purified by reverse phase preparative to give example 40(12mg, 28%). LCMS ESI (+) M/z 454.2(M +1).

Example 41(2185)

1- (9-fluoro-6, 11-dihydrobenzo [5,6] azepino [3,2,1-hi ] indazol-6-yl) -5-hydroxy-3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione

Step A: methyl 1H-indazole-7-carboxylic acid methyl ester (Compound 41.1)

Methyl 2-amino-3-methylbenzoate (2g, 12.18mmol) was dissolved in 20mL of acetic acid solution, 5mL of water was added, sodium nitrite (1.25g, 18.18mmol) was slowly added, and stirring was carried out at room temperature for 2 hours. The reaction was quenched by addition of saturated sodium bicarbonate solution, extracted three times with 100mL ethyl acetate, the organic phases combined, spun dried and purified to give the product 41.1(640mg, 30%). LCMS ESI (+) m/z:177.1(M +1).

And B:1- (3-Fluorobenzyl) -1H-indazole-7-carboxylic acid methyl ester (Compound 41.2)

Compound 41.1(640mg, 3.6mmol) was dissolved in 20mL of N, N-dimethylformamide, 3-fluorobenzyl bromide (1.02g, 5.4mmol), cesium carbonate (2.3g, 7.23mmol) and nitrogen were added, and the reaction mixture was stirred at 80 ℃ for 2 hours. The reaction was quenched by addition of aqueous solution, extracted three times with 100ml ethyl acetate, the organic phases combined, spun dried and purified to give product 41.2(700mg, 68%). LCMS ESI (+) m/z:285.1(M +1)

And C:1- (3-Fluorobenzyl) -1H-indazole-7-carboxylic acid (Compound 41.3)

Compound 41.2(700mg, 2.5mmol) was dissolved in 10mL of methanol and 10mL of an aqueous solution, lithium hydroxide (310mg, 7.4mmol) was added, and the reaction mixture was stirred at room temperature overnight. The reaction solution was adjusted to pH 5 with HCl (2M), a solid precipitated, filtered and the solid dried to give 41.3(580mg, 87%). LCMS ESI (+) m/z: 271.1(M +1)

Step D: 9-Fluorobenzo [5,6] azabenzo [3,2,1-hi ] indazol-6 (11H) -one (Compound 41.4)

Compound 41.3(580mg, 2.1mmol) was charged into a 100mL round-bottom flask, 20mL PPA was added, and the reaction was stirred at 120 ℃ for 3 hours. Cooled to room temperature, quenched with 50ml of water, extracted three times with 100ml of ethyl acetate, the organic phases combined, spun dry and purified to give 41.4(530mg, 98%). LCMS ESI (+) m/z: 253.1(M +1).

Step E: 9-fluoro-6, 11-dihydrobenzo [5,6] azepino [3,2,1-hi ] indazol-6-ol (Compound 41.5)

Compound 41.4(530mg, 2.1mmol) was dissolved in 10mL tetrahydrofuran and 2mL methanol, the reaction was cooled to 0 deg.C, sodium borohydride (100mg, 2.5mmol) was added in portions, stirred at room temperature for 0.5 h, quenched by addition of saturated aqueous ammonium chloride, extracted three times with 100mL ethyl acetate, the organic phases were combined, spun dry and purified to give product 41.5(400mg, 75%). LCMS ESI (+) m/z: 255.1 (M +1).

Step F:5- (benzyloxy) -1- (9-fluoro-6, 11-dihydrobenzo [5,6] azepino [3,2,1-hi ] indazol-6-yl) -3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione

Compound 41.5(54mg, 0.21mmol) was dissolved in 5mL of N, N-dimethylformamide, intermediate A (53mg, 0.18mmol) was added, followed by 5mL of 1-propylglycerophosphate (50% in DMF) solution, and the reaction was stirred at 100 ℃ for 2 hours. The reaction was quenched by addition of water (20ml), extracted three times with 100ml ethyl acetate, the organic phases combined, spun dry and purified to give product 41.6(60mg, 54%). LCMS ESI (+) m/z:535.2(M +1)

Step G: 1- (9-fluoro-6, 11-dihydrobenzo [5,6] azepino [3,2,1-hi ] indazol-6-yl) -5-hydroxy-3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione (Compound 41)

Compound 41.6(60mg, 0.1mmol) was dissolved in 10mL of N, N-dimethylformamide, lithium chloride (216mg, 5.6mmol) was added, the reaction mixture was stirred at 100 ℃ for 4 hours, water (20mL) was added to quench the reaction, extraction was carried out three times with 100mL of ethyl acetate, the organic phases were combined, spun dry and purified to give example 41(27mg, 54%). LCMS ESI (+) m/z:445.2(M +1).

Referring to example 41, intermediate A was replaced by intermediate B (C, D, E, F), which was reacted with compound 41.5 separately in two steps to afford examples 42-45

Example 42(2193)

1' - (9-fluoro-6, 11-dihydrobenzo [5,6] azepino [3,2,1-hi ] indazol-6-yl) -5' -hydroxy-1 ', 2' -dihydrospiro [ cyclobutane-1, 3' -pyrido [1,2-b ] pyridazine ] -4', 6' -dione

LCMS ESI(+)m/z：457.2(M+1).

Example 43(2194)

1' - (9-fluoro-6, 11-dihydrobenzo [5,6] azepino [3,2,1-hi ] indazol-6-yl) -5' -hydroxy-1 ', 2' -dihydrospiro [ cyclopentane-1, 3' -pyrido [1,2-b ] pyridazine ] -4', 6' -dione

LCMS ESI(+)m/z：471.2(M+1).

Example 44(2197)

[1,2,4] triazine-6, 8-dione of (12AR) -12- (9-fluoro-6, 11-dihydrobenzo [5,6] azepino [3,2,1-hi ] indazol-6-yl) -7-hydroxy 3,4,12,12 a-tetrahydro-1H- [1,4] oxazino [3,4-C ] pyrido [2,1-f ]

LCMS ESI(+)m/z：474.2(M+1).

Example 45(2199)

[1,2,4] triazine-4, 6-dione of 1- (9-fluoro-6, 11-dihydrobenzo [5,6] azepino [3,2,1-hi ] indazol-6-yl) -5-hydroxy-3- ((R) -1,1, 1-trifluoropropan-2-yl) -2, 3-dihydro-1H-pyrido [2,1-f ]

LCMS ESI(+)m/z：514.1(M+1).

Example 46(2303)

1' - (9, 10-difluoro-6, 11-dihydrobenzo [5,6] azepino [3,2,1-hi ] indazol-6-yl) -5' -hydroxy-1 ', 2' -dihydrospiro [ cyclobutane-1, 3' -pyrido [1,2-b ] pyridazine ] -4', 6' -dione

The specific reaction equation is as follows:

step A methyl 1- (2, 3-difluorobenzyl) -1H-indazole-7-carboxylate (Compound 46.1)

1- (bromomethyl) -2, 3-difluorobenzene (1.6g, 7.7mmol) was dissolved in 20mL of N, N-dimethylformamide, and compound 41.1(920mg,5.2mmol) and cesium carbonate (3.4g,10.4mmol) were added at room temperature, heated to 60 ℃ and stirred for 1 hour. The reaction mixture was poured into 200mL of water, extracted three times with 30mL of ethyl acetate, and the organic phase was washed with saturated brine and dried over anhydrous sodium sulfate. Filtration, spin-drying and column-purification of the residue gave 46.1(760mg, yellow solid, yield: 48.1%). LCMS ESI (+) M/z 303.1(M +1).

Step B1- (2, 3-difluorobenzyl) -1H-indazole-7-carboxylic acid (compound 46.2)

Compound 46.1(760mg, 2.50mmol) was dissolved in 10mL of tetrahydrofuran solution, and 10mL of aqueous sodium hydroxide (3M) was added at room temperature, heated to 70 ℃ and stirred overnight. The reaction mixture was neutralized with concentrated hydrochloric acid to pH 5, and then extracted twice with 20mL of ethyl acetate, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and then dried by spin-drying to give compound 46.2(700mg, 98%) as a white solid. LCMS ESI (+) M/z 289.1(M +1).

Step C9, 10-Difluorobenzo [5,6] azabicyclo [3,2,1-H i ] indazol-6 (11H) -one (Compound 46.3)

Compound 46.2(200mg,0.69mmol) was dissolved in 2mL of polyphosphoric acid and 4mL of sulfolane at 100 ℃ and stirred for reaction for 16 hours. Water (40mL) was added to the reaction, extracted three times with ethyl acetate (20mL), and washed 1 time with sodium bicarbonate (20 mL). Dry over anhydrous sodium sulfate and spin dry to give 46.3(80mg, 42.7%) as a yellow solid. LCMSISI (+) M/z 267.1(M +1).

Step D9, 10-difluoro-6, 11-dihydrobenzo [5,6] azabicyclo [3,2, 1-xipin ] indazol-6-ol (Compound 46.4)

Under ice-bath conditions, compound 46.3(80mg,0.3mmol) was dissolved in tetrahydrofuran/methanol (5mL/1mL), followed by addition of sodium borohydride (21mg,0.55mmol) and stirring was continued at zero degrees for 1 hour. The mixture was quenched by addition of saturated ammonium chloride solution (5mL), extracted 2 times with ethyl acetate (15mL), and washed 1 time with saturated brine (20 mL). Concentration by drying gave compound 46.4(80mg, 99%) as a white solid. LCMS ESI (+) M/z 273.1(M +1).

Step E5 ' - (benzyloxy) -1' - (9, 10-difluoro-6, 11-dihydrobenzo [5,6] azepino [3,2,1-hi ] indazol-6-yl) -1', 2' -dihydrospiro [ cyclobutane-1, 3' -pyrido [1,2-b ] pyridazine ] -4', 6' -dione (Compound 46.5)

Intermediate B (130mg, 0.42mmol) was dissolved in 1mL of N, N-dimethylformamide, compound 46.4(40mg, 0.15mmol) and 3mL of 1-propylphosphoric anhydride (50% in DMF) solution were added, and the reaction was stirred at 100 ℃ for 1 hour. The reaction was quenched with saturated sodium bicarbonate solution (5mL), and the aqueous phase was extracted 2 times with ethyl acetate (15mL) and dried over anhydrous sodium sulfate. Filtration, spin-drying and purification of the residue yielded compound 46.5(47mg, yellow solid, 41.8%). LCMS ESI (+) M/z:565.2(M +1)

Step F1 ' - (9, 10-difluoro-6, 11-dihydrobenzo [5,6] azepino [3,2,1-hi ] indazol-6-yl) -5' -hydroxy-1 ', 2' -dihydrospiro [ cyclobutane-1, 3' -pyrido [1,2-b ] pyridazine ] -4', 6' -dione (Compound 46)

Compound 46.5(47mg, 0.083mmol) was dissolved in 3mL of N, N-dimethylformamide, lithium chloride (30mg, 0.71mmol) was added, and the mixture was heated to 100 ℃ for 1 h. 10mL of water and 10mL of ethyl acetate were added, the layers were separated with stirring, the aqueous layer was extracted with ethyl acetate (10mL X3), and the organic layers were combined, washed with saturated brine (30mL) and dried over anhydrous sodium sulfate. Filtration, spin-drying, purification by reverse phase preparative and lyophilization afforded example 46(21mg, 26.3%) as a yellow solid. LCMS ESI (+) M/z 475.2(M +1).

Reference example 46 intermediate B was replaced by intermediate A (C, D), which was reacted with compound 46.5 separately, to give a two-step synthesis

Examples 47 to 49

Example 47(2302)

1- (9, 10-difluoro-6, 11-dihydrobenzo [5,6] azepino [3,2,1-hi ] indazol-6-yl) -5-hydroxy-3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione

LCMS ESI(+)m/z:463.2(M+1)

Example 48(2304)

1' - (9, 10-difluoro-6, 11-dihydrobenzo [5,6] azepino [3,2,1-hi ] indazol-6-yl) -5' -hydroxy-1 ', 2' -dihydrospiro [ cyclopentane-1, 3' -pyrido [1,2-b ] pyridazine ] -4', 6' -dione

LCMS ESI(+)m/z:489.2(M+1)

Example 49(2308)

[1,2,4] triazine-6, 8-dione of (12AR) -12- (9, 10-difluoro-6, 11-dihydrobenzo [5,6] azepino [3,2, 1-xi ] indazol-6-yl) -7-hydroxy 3,4,12,12 a-tetrahydro-1H- [1,4] oxazino [3,4-C ] pyrido [2,1-f ]

LCMS ESI(+)m/z:492.1(M+1)

Example 50(2311)

(S) -1- (2-fluoro-5, 11-dihydrobenzo [6,7] thiaheptacyclo [3,4-b ] pyridin-5-yl) -5-hydroxy-3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione

Step A: (S) -5- (benzyloxy) -1- (2-fluoro-5, 11-dihydrobenzo [6,7] thiaheptacyclo [3,4-b ] pyridin-5-yl) -3, 3-dimethyl-2, -1, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione

Compound 19.3(200mg,0.38mmol) was subjected to chiral separation to give compound 50.1(90mg, 45%). LCMS ESI (+) M/z 528.2(M +1).

Chiral separation conditions:

chiral column ChiralCel OD-H (Daicel chemical Industries, Ltd,250 x 30mm i.d.,5 um); mobile phase A is supercritical CO2, mobile phase B is isopropanol (A: B: 60: 40); flow rate: 50 ml/min; the column temperature is 38 ℃; detection wavelength of 220nm

Step B (S) -1- (2-fluoro-5, 11-dihydrobenzo [6,7] thiaheptacyclo [3,4-B ] pyridin-5-yl) -5-hydroxy-3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-B ] pyridazine-4, 6-dione (Compound 50)

Compound 50.1(50mg,0.095mmol) was dissolved in NN-dimethylformamide (0.5mL), and lithium chloride (21mg, 0.48mmol) was added thereto, followed by stirring at 100 ℃ for 5 hours. The solution was cooled to room temperature and purified directly by reverse phase preparative lyophilization to give example 50(22 mg, 53%). LCMS ESI (+) M/z 438.1(M +1).

Referring to the isolation procedure of example 50, isolation preparations gave examples 51-60, as shown in the following table:

example 61 (2115):

5-hydroxy-3-isopropyl-1- (2-methyl-2, 6-dihydrobenzo [6,7] thiaheptacyclo [2,3,4-c, d ] indazol-6-yl) -2, 3-dihydro-1H-pyrido [1,2-f ] [1,2,3] triazine-4, 6-dione

Referring to example 2, the same procedure was used to replace intermediate A with intermediate F to give example 61.LCMS ESI (+) M/z:474.2(M +1)

Example 62 (2133):

[1,2,4] triazine-4, 6-dione of 1- (6, 11-dihydro-1, 2,11 a-triazabenzo [ cd, g ] -azulen-6-yl-5-hydroxy-3- ((R) -1,1, 1-trifluoropropanidin-2-yl) -2, 3-dihydro-1H-pyrido [2,1-f ]

Referring to example 1, intermediate A was replaced with intermediate E and the same procedure was used to give example 62.LCMS ESI (+) M/z:497.1 (M +1).

Example 63 (2165):

(R) -12- ((S) -6, 12-dihydrobenzo [6',7' ] thiaheptacyclo [3',4':3,4] benzo [1,2-C ] [1,2,5] thiadiazol-6-yl) -7-hydroxy-3, 4,12,12 a-tetrahydro-1H- [1,4] pyrano [4,3-C ] pyrido [2,1-f ] [1,2,4] triazine-6, 8(1H, 3H) -dione

Referring to example 6, the same procedure was used to replace intermediate A with intermediate D to give example 63.LCMS ESI (+) M/z:506.1(M +1)

Example 64

[1,2,4] triazine-4, 6-dione of 5-hydroxy-3-isopropyl-1- (4-methyl-4, 6,7,12 tetrahydrobenzo [5,6] cycloocta [1,2,3-cd ] indol-12-yl) -2, 3-dihydro-1H-pyrido [2,1-f ]

The specific synthesis steps are as follows:

step A: 1-methyl-1H-indole-4-carboxylic acid methyl ester (Compound 64.1)

The compound methyl-1H-indole-4-carboxylic acid methyl ester (4g, 22.86mmol) was dissolved in a solution of N, N-dimethylformamide (20mL), sodium hydrogen (3.65g, 91.43mmol) was slowly added under ice bath and stirred for 30 minutes, iodomethane (5.7mL, 91.43mmol) was further added, and stirred at room temperature for two hours. The reaction was quenched by addition of saturated aqueous ammonium chloride solution, extracted three times with 100mL ethyl acetate, the organic phases combined, spun dry and purified to give the product 64.1(4g, 92%). LCMS ESI (+) m/z: 190(M +1).

And B: 3-formyl-1-methyl-1H-indole-4-carboxylic acid methyl ester (Compound 64.2)

Phosphorus oxychloride (2.4mL, 25.40mmol) was slowly added to a solution of N, N-dimethylformamide (10mL) under ice-bath, the resulting mixture was stirred at zero degrees centigrade for 30 minutes, a solution of compound 64.1(4g, 21.16mmol) in N, N-dimethylformamide was added to the reaction solution, and the mixture was stirred for 6 hours. The reaction mixture was poured into an ice-water mixture and the pH was adjusted to 7 by adding sodium hydroxide solution. Three extractions with 100mL ethyl acetate were performed and the organic phases were combined. Spin dry and purify to give the product (4.2g, 91%). LCMS ESI (+) m/z: 218(M +1).

And C: (Z) -1-methyl-3-styryl-1H-indole-4-carboxylic acid methyl ester (Compound 64.3)

Triphenylphosphine bromide (19.6g, 45.2mmol) was dissolved in 25mL tetrahydrofuran, n-butyllithium (27mL, 67.74mmol) was added dropwise at-78 deg.C under nitrogen, and the reaction was stirred for 0.5 h. A tetrahydrofuran solution of compound 64.2(4.2g, 19.35mmol) was added dropwise thereto, and the reaction mixture was warmed to room temperature and stirred for two hours. The reaction was quenched by addition of saturated aqueous ammonium chloride, extracted three times with 100mL ethyl acetate, and the organic phases combined. Spin-dry and purify to give 64.3(3.6g, 55%). LCMS ESI (+) m/z: 292(M +1).

Step D: 1-methyl-3-phenylethyl-1H-indole-4-carboxylic acid methyl ester (compound 64.4)

Compound 64.3(3.6g,12.54mmol) was dissolved in 40mL of methanol solution, palladium on carbon (245mg) was added, and the reaction was stirred under hydrogen for 4 hours. Filtration through celite and spin-drying of the organic phase purified to give product 64.4(3.2g, 88%). LCMSISI (+) m/z: 294(M +1).

Step E: 1-methyl-3-phenylethyl-1H-indole-4-carboxylic acid (compound 64.5)

Compound 64.4(3.2g, 10.92mmol) was dissolved in 20mL of methanol solution, and aqueous sodium hydroxide (6M, 10mL) was added and stirred at room temperature for two hours. Ph was adjusted to 5 by addition of 3M aqueous hydrochloric acid and the solid was filtered and the solvent removed to give product 64.5(2.5g, 82%). LCMS ESI (+) m/z:280 (M +1).

Step F: 4-methyl-6, 7-dihydrobenzo [5,6] cycloocta [1,2,3-cd ] indole 12(4H) -one (Compound 64.6)

Compound 64.5(2.5g, 8.96mmol) was dissolved in 20mL of dichloromethane, oxalyl chloride (1.9mL, 22.4mmol) was added dropwise at zero degrees Celsius, the mixture was warmed to room temperature and stirred for 4 hours, and the solvent was directly dried by rotary evaporation to remove excess oxalyl chloride. Dissolve in dichloromethane solution, add aluminum trichloride (3.6g, 26.88mmol) at zero degrees Celsius, stir for 2 hours, filter through celite, spin dry the solvent, and purify to give 64.6(350mg, 15%). LCMS ESI (+) m/z:262 (M +1).

Step G: 4-methyl-4, 6,7, 12-tetrahydrobenzo [5,6] cycloocta [1,2,3-cd ] indol-12-ol (compound 64.7)

Compound 64.6(350mg, 1.34mmol) was dissolved in 10mL of methanol, and sodium borohydride (61mg, 1.61mmol) was added in portions at zero degrees Celsius, and stirred at room temperature for two hours. The reaction was quenched by addition of saturated aqueous ammonium chloride, extracted three times with 30mL ethyl acetate, the organic phases combined, spun dried and purified to give the product 64.7(350mg, 98%). LCMS ESI (+) m/z:264 (M +1).

Step H: [1,2,4] triazine-4, 6-dione of 5- (benzyloxy) -3-isopropyl-1- (4-methyl-4, 6,7, 12-tetrahydrobenzo [5,6] cycloocta [1,2,3-cd ] indol-12-yl) -2-1, 3-dihydro-1H-pyrido [2,1-f ] (Compound 64.8)

Compound 64.7(150mg, 0.57mmol) was dissolved in 5mL of 1, 2-dichloroethane solution, and intermediate F (214mg, 0.68mmol) and dichloroacetic acid (0.12mL, 1.42mmol) were added, and the mixture was heated under reflux overnight under nitrogen. The reaction was quenched by the addition of saturated aqueous sodium bicarbonate solution, extracted three times with 30mL of dichloromethane, the organic phases combined, spun dried and purified to give the product 64.8(150mg, 47%). LCMS ESI (+) m/z: 559(M +1).

Step I: [1,2,4] triazine-4, 6-dione of 5-hydroxy-3-isopropyl-1- (4-methyl-4, 6,7, 12-tetrahydrobenzo [5,6] cycloocta [1,2,3-cd ] indol-12-yl) -2, 3-dihydro-1H-pyrido [2,1-f ] (example 64)

Compound 64.8(150mg, 0.269mmol) was dissolved in 5mL of N, N-dimethylformamide, and lithium chloride (90mg, 2.2mmol) was added. The reaction solution was stirred at 100 ℃ for 6 hours under nitrogen protection. 20mL of water was added, extracted three times with 30mL of ethyl acetate, the organic phases were combined, spun dry and purified to give the product, example 64(50mg, 40%). LCMS ESI (+) M/z 469.2(M +1).

Example 65

[1,2,4] triazine-4, 6-dione of 5-hydroxy-3-isopropyl-1- (2-methyl-6, 11-dihydro-2H-benzo [5,6] cyclohepta [1,2,3-cd ] indol-6-yl) -2, 3-dihydro-1H-pyrido [2,1-f ]

Step A: compound 3-benzyl-1H-indole-4-carboxylic acid methyl ester (Compound 65.1)

The weighed compound methyl-1H-indole-4-carboxylate (525mg, 3mmol) was dissolved in dichloromethane (12mL), and then benzaldehyde (477mg, 4.5mmol), triethylsilane (1.15g, 9.9mmol) and trifluoroacetic acid (787mg, 6.9mmol) were added in this order under ice bath, slowly warmed to room temperature and stirred for 1 hour. The reaction was confirmed to be complete by LCMS. The reaction was quenched with saturated sodium bicarbonate solution (15mL), the layers were separated, the aqueous layer was extracted with dichloromethane (12mL), the combined organic layers were washed with saturated brine (20mL) and dried over anhydrous sodium sulfate. Filtration, spin-drying and purification gave 65.1(590mg, yield: 74.2%). LCMS ESI (+) M/z 266.1(M +1).

And B: 3-benzyl-1-methyl-1H-indole-4-carboxylic acid methyl ester (Compound 65.2)

After compound 65.1(590mg, 2.23mmol) was dissolved in N, N-dimethylformamide (8mL) under ice-bath conditions, 60% sodium hydride (134mg, 3.35mmol) was slowly added thereto, stirring was continued for 30 minutes, methyl iodide (412mg, 2.9mmol) was added to the reaction flask, and the reaction was stirred at room temperature for 1 hour. TLC and LCMS confirmed the reaction was complete. The reaction was quenched with saturated ammonium chloride solution (10mL), extracted with ethyl acetate (12mL), and the organic phase was washed with saturated brine (20mL) and dried over anhydrous sodium sulfate. Filtration, spin-drying and purification gave 65.2(500mg, yield: 80.49%). LCMS ESI (+) M/z 280.1(M +1).

And C: 3-benzyl-1H-indole-4-carboxylic acid (Compound 65.3)

Compound 65.2(500mg, 1.79mmol) was dissolved in tetrahydrofuran/methanol (10mL, 5:1), and sodium hydroxide solution (6M,2.5mL) was added to the reaction solution, which was stirred at 70 ℃ for 2 hours. TLC and LCMS confirmed the reaction was complete. The reaction solution was concentrated, water (10mL) was added, and then acidified to pH 3 with a 2M hydrochloric acid solution, filtered, and spun-dried to obtain 65.3(450mg, yield: 94.7%) as a white solid. LCMS ESI (+) M/z 266.1(M +1).

Step D: 3-benzyl-1-methyl-1H-indole-4-carbonyl chloride (Compound 65.4)

After 65.4(450mg, 1.7mmol) was dissolved in dry dichloromethane (6mL) under ice-bath conditions, oxalyl chloride (626mg, 4.93mmol) was slowly dropped into the reaction flask, and the reaction was stirred at room temperature for 1 hour. Completion of the reaction was confirmed by TLC. The reaction was then directly spin dried to give crude compound 65.4.

Step E: 2-methyl-2H-benzo [5,6] cyclohepta [1,2,3-cd ] indol-6 (11H) -one (compound 65.5)

Aluminum trichloride (452mg, 3.4mmol) was dissolved in dry dichloromethane (6mL) under ice-bath conditions, and then a solution of compound 65.4 in dichloromethane (6mL) was slowly dropped into the reaction flask, warmed to 50 ℃ and stirred for reaction for 30 minutes. The reaction was confirmed to be complete by TLC. The reaction was quenched with water (10mL) at room temperature, the reaction was extracted with ethyl acetate (12mL), and the combined organic phases were washed with saturated brine (20mL) and dried over anhydrous sodium sulfate. Filtration, concentration and purification gave 65.5(250mg, yield: 59.6%).

Step F: 2-methyl-2H-benzo [5,6] cyclohepta [,2,3-cd ] indole-6 (11H) -ol (Compound 65.6)

Compound 65.5(250mg, 1.01mmol) was dissolved in tetrahydrofuran/methanol (10mL, 5:1), sodium borohydride (46mg, 1.21mmol) was added to the reaction flask under ice bath, and the reaction was stirred at room temperature for 1 hour. The reaction was confirmed to be complete by TLC and LCMS. After quenching the reaction with a saturated ammonium chloride solution, water (10mL) was added to dissolve the precipitated salt, followed by extraction with ethyl acetate (12mL), and the organic phases were combined, washed with saturated brine, and dried over anhydrous sodium sulfate. Filtration, spin-drying and purification gave 65.6 as a pale yellow solid (250mg, yield: 99.2%). LCMS ESI (+) M/z:232.1(M-17).

Step G: [1,2,4] triazine-4, 6-dione of 5- (benzyloxy) -3-isopropyl-1- (2-methyl-6, 11-dihydro-2H-benzo [5,6] cyclohepta [1,2,3-cd ] indol-6-yl) -2, -1, 3-dihydro-1H-pyrido [2,1-f ] (Compound 65.7)

Intermediate F (100mg, 0.32mmol) and compound 65.6(96mg, 0.384mmol) were dissolved in 1, 2-dichloroethane (8mL) and dichloroacetic acid (2 drops) was added and the reaction was stirred overnight in an oil bath at 85 ℃. LCMS confirmed reaction complete. Water (12mL) was added to the reaction flask, and the mixture was extracted with ethyl acetate (12mLX3), washed with saturated brine, and the organic phase was dried over anhydrous sodium sulfate. Filtration and concentration gave 65.7(185mg) as a crude pale yellow oil. LCMS ESI (+) M/z 545.2(M +1).

Step H: [1,2,4] triazine-4, 6-dione of 5-hydroxy-3-isopropyl-1- (2-methyl-6, 11-dihydro-2H-benzo [5,6] cyclohepta [1,2,3-cd ] indol-6-yl) -2, 3-dihydro-1H-pyrido [2,1-f ] (example 65)

Compound 65.7(185mg, 0.34mmol) was dissolved in 8mL of N, N-dimethylformamide and weighed amount of lithium chloride (228.5mg, 5.44mmol) was added to the reaction flask and the reaction was stirred in a 100 deg.C oil bath for 3 hours. The reaction was confirmed to be complete by LCMS. Water (15mL) was added to the reaction flask, and the mixture was extracted with ethyl acetate (10mL), and the organic phase was washed with saturated brine and dried over anhydrous sodium sulfate. Filtration, concentration and purification by reverse phase preparative gave example 65(25mg, yield: 16.23%) as a pale yellow solid. LCMS ESI (+) M/z 455.2(M +1).

Example 66

[1,2,4] triazine-4, 6-dione of 5-hydroxy-1- (2-methyl-6, 11-dihydro-2H-benzo [5,6] cyclohepta [1,2,3-cd ] indol-6-yl) -3- ((R) -1-1, 1-trifluoropropan-2-yl) -2, 3-dihydro-1H-pyrido [2,1-f ]

The specific synthesis steps are as follows:

step A: [1,2,4] triazine-4, 6-dione of 5- (benzyloxy) -1- (2-methyl-6, 11-dihydro-2H-benzo [5,6] cyclohepta [1,2,3-cd ] indol-6-yl) -3- ((R) -1,1, 1-trifluoropropan-2-yl) -2, 3-dihydro-1H-pyrido [2,1-f ] (Compound 66.1)

Intermediate E (50mg, 0.14mmol) and Compound 66.1(42mg, 0.168mmol) were dissolved in 8mL of 1, 2-dichloroethane solution, a catalytic amount of dichloroacetic acid was added, and the reaction was stirred overnight in an oil bath at 85 deg.C. LCMS confirmed reaction complete. The reaction was quenched by addition of water (12mL), extracted with ethyl acetate (12mL), and the organic phase was washed with saturated brine and dried over anhydrous sodium sulfate. Filtration and purification by column chromatography gave 66.1(60mg, yield: 74.1%) as a pale yellow oil. LCMSISI (+) M/z 599.2(M +1).

And B: [1,2,4] triazine-4, 6-dione of 5-hydroxy-1- (2-methyl-6, 11-dihydro-2H-benzo [5,6] cyclohepta [1,2,3-cd ] indol-6-yl) -3- ((R) -1-, 1, 1-trifluoropropan-2-yl) -2, 3-dihydro-1H-pyrido [2,1-f ] (example 66)

Compound 66.1(60mg, 0.1mmol) was dissolved in 4mL of N, N-dimethylformamide, then weighed amount of lithium chloride (84mg, 2mmol) was added to the reaction flask and the reaction was stirred in a 100 deg.C oil bath for 6 hours. The reaction was confirmed to be complete by LCMS. The reaction was quenched by addition of water (15mL), then extracted with ethyl acetate (10mL), and the organic phase was washed with saturated brine and dried over anhydrous sodium sulfate. Filtration, spin-drying, and purification by reverse phase preparative purification gave example 66(10mg, yield: 19.6%) as a pale yellow solid. LCMS ESI (+) M/z 509.2(M +1).

Example 67

(S) -1- (6,7,8,9-d 4-2-fluoro-5, 11-dihydrobenzo [6,7] thiaheptacyclo [3,4-b ] pyridin-5-yl) -5-hydroxy-3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione

The specific synthesis steps are as follows:

step A: 2-bromomethyl-6-chloro-nicotinic acid methyl ester (67.2)

The compound 2-methyl-6-chloro-nicotinic acid methyl ester (67.1, 20mmol), NBS (22mmol), and perbenzoic acid anhydride (1mmol) were refluxed in tetrachloromethane (50mL) until all starting material disappeared. The reaction was concentrated to about 10mL and separated on a silica gel column (10-50% ethyl acetate/hexanes) to provide compound 67.2.

And B: 6-chloro-2- (((phenyl-d 5) thio) methyl) nicotinic acid methyl ester (67.3)

Compound 67.2(15mmol), (phenyl-d 5) thiol (16mmol) and potassium carbonate (20mmol) was stirred in acetonitrile (50mL) overnight. The reaction was filtered and concentrated, leaving the mixture to be purified on silica gel to give 67.3.

And C: 2-chlorobenzo [6,7] thiaheptacyclo [3,4-b ] pyridin-5 (11H) -one-6, 7,8,9-d4(67.4)

Compound 67.3(10mmol) was dissolved in 20mL of polyphosphoric acid, heated to 120 ℃ and stirred for 8 hours. TLC monitored the reaction complete. After cooling to room temperature, the reaction mixture was diluted with water (30mL), extracted with ethyl acetate (25mLX3), and washed with saturated sodium bicarbonate solution (20 mL). The organic phase was dried over anhydrous sodium sulfate, filtered and spin dried to give crude 67.4.

Step D: 2-fluoro-benzo [6,7] thiaheptacyclo [3,4-b ] pyridin-5 (11H) -one-6, 7,8,9-d4(67.4)

Compound 67.4, cesium fluoride (5 equivalents) was dissolved in DMSO, heated to 120 ℃ and stirred until TLC monitored for reaction completion. After cooling to room temperature, the reaction mixture was diluted with water, extracted with ethyl acetate (25mLX3), and washed with saturated sodium bicarbonate solution (20 mL). The organic phase is dried by anhydrous sodium sulfate, filtered and dried by spinning to obtain a crude product, and the crude product is separated and purified by a silica gel column to obtain 67.5.

Step E: 2-fluoro-benzo [6,7] thiaheptacyclo [3,4-b ] pyridin-5-ol-6, 7,8,9-d4(67.5)

Compound 67.4 was dissolved in tetrahydrofuran/methanol (5:1), sodium borohydride (1.0 eq) was added to the reaction flask while on ice bath, and the reaction was stirred at ambient temperature for 1 hour. The reaction was confirmed to be complete by TLC and LCMS. After quenching the reaction with ammonium chloride solution, extraction was performed with ethyl acetate, and the organic phases were combined, washed with saturated brine, and dried over anhydrous sodium sulfate. Filtering, spin-drying, and purifying with silica gel column to obtain 67.5.

Step E: (S) -1- (6,7,8,9-d 4-2-fluoro-5, 11-dihydrobenzo [6,7] thiaheptacyclo [3,4-b ] pyridin-5-yl) -5-hydroxy-3, 3-dimethyl-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione

The procedure used to prepare example 50 was used to prepare compound 67.5 to prepare example 67.

Example 68

1- (9, 10-difluoro-6, 11-dihydrobenzo [5,6] azepino [3,2,1-hi ] indazol-6-yl) -5-hydroxy-3, 3-dimethyl-d 6-2, 3-dihydro-1H-pyrido [1,2-b ] pyridazine-4, 6-dione

The synthesis of intermediate a was used to synthesize intermediate a2, in which deuterated isobutyraldehyde was substituted for isobutyraldehyde. The same synthetic method of example 53 was used to synthesize example 68.

Example 69

(S) -1' - (9, 10-difluoro-6, 11-dihydrobenzo [5,6] azepino [3,2,1-hi ] indazole-11, 11-d 2-16-yl) -5' -hydroxy-1 ', 2' -dihydrospiro [ cyclopentane-1, 3' -pyrido [1,2-b ] pyridazine ] -4', 6' -dione

Step A: (2, 3-difluorophenyl) methanol-1, 1-d2(69.2)

Methyl 2, 3-difluorobenzoate (10mmol) was dissolved in tetrahydrofuran (50mL), frozen to 0 deg.C, and lithium aluminum deuterium (10mmol) was added to the solution. After half an hour, the reaction was quenched with ammonium chloride solution, extracted with ethyl acetate, the organic phases combined and washed with saturated brine and dried over anhydrous sodium sulfate. Filtering and spin-drying to obtain 69.2

And B:1 (bromomethyl-d 2) -2, 3-difluorobenzene (69.3)

Adding dibromosulfoxide into dichloromethane solution of the compound 69.2, reacting for 2 hours at room temperature, concentrating to dryness, and separating with silica gel column to obtain 69.3

Step C methyl 1- ((2, 3-difluorophenyl) methyl-d 2) -1H-indazole-7-carboxylate (compound 69.4)

Compound 69.3 was dissolved in N, N-dimethylformamide, and compound 1H-indazole-7-carboxylic acid methyl ester and cesium carbonate were added at room temperature, heated to 60 ℃, and stirred for 1 hour. The reaction mixture was poured into water, extracted three times with ethyl acetate, and the organic phase was washed with saturated brine and dried over anhydrous sodium sulfate. Filtering, spin-drying, and purifying the residue by column chromatography to obtain compound 69.4.

The same synthetic method as for example 58 was used to prepare compound 69.4 as in example 69

Example 70: inhibiting viral proliferative Activity (EC)₅₀) And Cytotoxicity (CC) of the Compound₅₀)

1) Cell treatment: MDCK cells were seeded at a certain density in 384-well plates, and then the cells were placed at 37 ℃ in 5% CO₂The culture was carried out overnight in an incubator.

2) Compound treatment: the test compound will be diluted in DMSO at a multiple ratio and added to the cell culture plate.

3) Virus inoculation: diluted influenza A/Weiss/43(H1N1) virus solution was then added to the antiviral activity assay wells, and no virus was added to the cytotoxicity assay wells. The final concentration of DMSO was 0.5%. The cells were incubated at 37 ℃ with 5% CO₂Incubate for 5 days in an incubator until the rate of cellular disease in virus control wells (no compound) reaches 80-95%.

4) And (3) detecting the activity of the cells: adding cell activity detection reagent CCK-8 detection reagent into each well, and detecting at 37 deg.C and 5% CO₂After culturing the cells in the incubator for 3-4 hours, detecting a signal value by using a spectrophotometer, and calculating the anti-influenza virus activity and cytotoxicity of the compound by using the original data. Dose response curves of compounds and their EC₅₀And CC₅₀Values will be obtained after analysis by GraphPad Prism software. .

The method can be used to determine the activity (EC) for inhibiting virus proliferation₅₀) And Cytotoxicity (CC) of the Compound₅₀) Are shown in Table

TABLE-Activity and cytotoxicity test results

Claims

1. An azacyclodione compound, which is characterized by being a compound represented by the following structure:

wherein the content of the first and second substances,

R¹is selected fromHydrogen, halogen, cyano, optionally substituted alkyl, amino, optionally substituted alkoxy;

R²selected from hydrogen, -C (═ O) R^A,-C(＝O)OR^A,-CH₂OC(＝O)R^A,-CH₂OC(＝O)OR^A,-CH(Me)OC(＝O)R^A,-CH(Me)OC(＝O)OR^A；

R^ASelected from alkyl, substituted alkyl;

x is selected from NR⁴,CR⁵R⁶；

R³selected from hydrogen, alkyl, substituted alkyl;

or R³And R⁴Together form an optionally substituted heterocyclyl;

Y is selected from S, S (O), O, N, CR⁸R⁹；

ring C is selected from optionally substituted aryl;

R⁷hydrogen, halogen, cyano, alkyl, alkoxy, nitro, amino, amido and amido;

2. An azacyclic dione compound as claimed in claim 1, wherein:

said Q¹，Q²，Q³，Q⁴And Q⁵Two or more of them are nitrogen atoms.

3. The azacyclic dione compound of claim 1, which is a compound of the structure:

wherein R is¹，R²，R³Ring A is shown as general formula I;

m is 0, 1,2,3, 4;

R⁴is hydrogen optionally substituted on the polycyclic ring, alkyl, substituted alkyl, optionally substituted alkoxy, optionally substituted amino, optionally substituted heterocyclic radical and optionally substituted cycloalkyl.

4. The azacyclic dione compound of claim 1, which is a compound of the structure:

wherein, X, Y, R²，R³R7, n, ring C are shown as general formula I;

R₇₁and R₇₂Forming a five-membered aromatic heterocycle; orR₇₃And R₇₂Forming a five-membered heteroaromatic ring.

5. The azacyclic dione compound of claim 4, which is a compound of the structure:

wherein A, B, C, D, E, F is selected from nitrogen, sulfur and oxygen;

6. The azacyclic dione compound of claim 1, which is a compound of the structure:

wherein, X, Y, R²，R³R7, n, ring C are shown as general formula I;

ring B is a five or six membered aromatic ring.

7. An azacyclic dione compound according to claim 6, wherein:

one carbon atom on the B ring is substituted by carbonyl and oxime group.

8. An azacyclic dione compound as claimed in claim 1, wherein:

the optionally substituted alkyl refers to that hydrogen atoms on an alkyl carbon chain are substituted by hydrogen, halogen, alkyl, nitro, amino, alkenyl, alkynyl, ester group, amido, optionally substituted cycloalkyl/heterocycloalkyl and optionally substituted aryl, or carbon atoms on an alkyl carbon chain are substituted by oxygen, sulfur, nitrogen and carbonyl;

the optionally substituted aryl refers to that hydrogen atoms on an aryl ring are substituted by hydrogen, halogen, alkyl, nitro, amino, alkenyl, alkynyl, ester group and amido, or carbon atoms on the aryl ring are substituted by carbonyl;

the optionally substituted heteroaryl refers to the replacement of hydrogen atoms on a heteroaryl ring by hydrogen, halogen, alkyl, nitro, amino, alkenyl, alkynyl, ester group and amide group, or the replacement of carbon atoms on the heteroaryl ring by carbonyl groups.

9. An azacyclic dione compound according to any one of claims 1 to 8, wherein:

one or more hydrogen atoms on the azacyclo-diketone compound are replaced by an isotope of hydrogen.

10. An azacyclic dione compound as claimed in any one of claims 1 to 9, wherein:

application to the inhibition of cap-dependent endonuclease activity.