Compounds having antiviral activity

Abstract

Provided in the present invention are novel compounds, a composition thereof, a preparation method therefor and the use thereof. The compounds (I) and the composition thereof of the present invention have a significant anti-human immunodeficiency virus effect, and have significantly improved in-vivo bioavailability.

Description

A compound with antiviral activity Technical Field

[0001] This invention generally relates to compounds for treating HIV infection, and more particularly to compositions and uses for treating HIV infection. Background Technology

[0002] Since the first case of Acquired Immunodeficiency Syndrome (AIDS) was discovered in 1981, tens of millions of new cases have been added each year, and the trend is increasing annually. Due to the lack of a specific treatment for AIDS, the vast majority of patients succumb to the disease. Currently, it is estimated that 200 million people worldwide are infected with Human Immunodeficiency Virus (HIV). AIDS is caused by HIV infection, which leads to a deficiency in the body's immune system, making individuals more susceptible to other related diseases.

[0003] To conquer AIDS, scientists worldwide have been dedicated to developing effective drugs for its treatment. Currently available drugs include the following: Nucleoside reverse transcriptase inhibitors (NRTIs) were the earliest developed antiviral agents, including zidovudine, zalcitabine, and lenacapavir. These drugs showed good anti-HIV effects in the early stages of clinical use, but significant drug resistance develops after more than six months of continuous use, and their effect on rebuilding the patient's immune system is poor. Non-nucleoside reverse transcriptase inhibitors (NNRTIs), such as nevirapine, deraviridine, and efavirenz, are mainly used in combination with other anti-HIV drugs. While they have improved resistance to drug resistance, they are prone to causing allergic reactions such as rashes. Since the 1990s, a number of protease inhibitors, such as saquinavir, ritonavir, and nelfinavir, have been marketed. These drugs can significantly inhibit viral replication and have shown strong anti-HIV-1 and HIV-2 activity in acutely infected lymphoid stem cells, while also reducing the incidence of adverse reactions. They have achieved good results in clinical use, but drug resistance has increased significantly. In 1996, scientists proposed "cocktail" therapy, a major milestone in the treatment of AIDS. This therapy can significantly reduce the amount of HIV in the patient's blood, demonstrating strong antiviral capabilities. This treatment regimen has been widely promoted and applied. However, this therapy requires intermittent administration, and viral rebound occurs after discontinuation of the drug; it can only control the viral load in the patient's blood below a certain limit.

[0004] Although highly effective antiretroviral therapy has played a significant role in the treatment of AIDS, the variability and drug resistance of HIV, coupled with the fact that new HIV strains require new antiretroviral drugs to eradicate them, necessitates the search for novel antiretroviral drugs. Summary of the Invention

[0005] This invention provides a novel compound and its composition, as well as its preparation method and uses.

[0006] In another aspect, the compounds and compositions provided by the present invention have significant antiviral effects against HIV-1 wild-type (IIIB) and HIV-2 (ROD) viruses both in vivo and in vitro, achieving significant beneficial technical effects.

[0007] A third aspect of the present invention is that the compounds provided by the present invention have higher blood drug concentrations, longer half-lives, and significantly improved bioavailability in vivo, achieving unexpected technical effects.

[0008] A fourth aspect of the present invention is that the compounds provided by the present invention have low toxicity in vivo, and it is expected that the compounds of the present invention will have fewer toxic side effects and have great potential for drug development.

[0009] This invention provides a compound of formula I:

[0010] Including its solvates, isotopic derivatives, or salts thereof;

[0011] in:

[0012] Y1 and Y2 are each independently selected from CH or N;

[0013] R1 and R2 are each independently selected from hydrogen, halogen, hydroxyl, cyano, and C. 1-6 Alkyl, C 1-6 Alkoxy;

[0014] Ring A is selected from five- to six-membered heterocycles having one to three nitrogen atoms and carbon atoms, and is optionally substituted with R3 or R4;

[0015] R3 and R4 are each independently selected from hydrogen, amino, hydroxyl, and C. 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 Carbocyclic group, deuterated C 1-6 Alkyl, -NHR9, -OR9, The aforementioned alkyl, alkoxy, and carbocyclic groups may optionally be substituted with one or more hydrogen, halogen, trifluoromethyl, cyano, methanesulfonyl, ethanesulfonyl, isopropanesulfonyl, cyclopropanesulfonyl, or p-toluenesulfonyl groups;

[0016] T is selected from hydrogen, sodium, potassium, Hydroxyethyl;

[0017] R w1 R w2 R w3 Each is independently selected from hydrogen and C. 1-6 alkyl;

[0018] R x R y Each is independently selected from hydrogen, metal ions, and C. 1-6 Alkyl, C 3-6 carbon cyclo group, C 2-10 Heterocyclic group, C 3-10 Heterocyclic aryl, or R x R y Linked into a ring; the above-mentioned heterocyclic groups and heterocyclic aryl groups may optionally be substituted by one or more hydrogens, halogens, hydroxyl groups, amino groups, nitro groups, methyl groups, ethyl groups, isopropyl groups, carboxyl groups, methoxy groups, ethoxy groups, isopropoxy groups, methylamino groups, ethylamino groups, isopropylamino groups, mercapto groups, mercaptomethyl groups, mercaptoethyl groups, acetyl groups, trifluoromethyl groups, methanesulfonyl groups, and ethanesulfonyl groups;

[0019] R z Selected from C 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 carbon cyclo group, C 2-10 Heterocyclic group, C 3-10 Heterocyclic aryl, deuterated C 1-6 Alkyl; the above-mentioned alkyl, alkoxy, carbocyclic, heterocyclic, and heterocyclic aryl groups may optionally be substituted by one or more hydrogen, halogen, hydroxyl, amino, nitro, methyl, ethyl, isopropyl, carboxyl, methoxy, ethoxy, isopropoxy, methylamino, ethylamino, isopropylamino, mercapto, mercaptomethyl, mercaptoethyl, acetyl, trifluoromethyl, methanesulfonyl, and ethanesulfonyl groups;

[0020] R5 is selected from hydrogen, halogen, hydroxyl, cyano, C 1-6 Alkyl, C 1-6 Alkoxy;

[0021] R6 is selected from C 1-6 Alkyl, C 1-6 Alkoxy, C 2-8 alkenyl, C 2-8 Alkyne group, deuterated C 2-8 Alkynyl; the above-mentioned alkyl, alkoxy, alkenyl, and alkynyl groups may optionally be substituted with one or more hydrogen, cyano, trifluoromethyl, methanesulfonyl, ethanesulfonyl, isopropanesulfonyl, cyclopropanesulfonyl, p-toluenesulfonyl, deuterated methanesulfonyl, deuterated ethanesulfonyl, deuterated isopropanesulfonyl, deuterated cyclopropanesulfonyl, deuterated formyl, or deuterated acetyl.

[0022] R7 is selected from C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 alkylamine group, C 3-6 Carbocyclic group; the above-mentioned alkyl, alkoxy, alkylamine, and carbocyclic groups may optionally be converted by one or more R groups. 10 replace;

[0023] R8 is selected from C 1-6 Alkyl, C 3-6 Carbocyclic group; the above-mentioned alkyl or carbocyclic group may optionally be converted by one or more R groups. 11 replace;

[0024] R9 is selected from formyl, acetyl, propionyl, cyclopropionyl, benzoyl, methanesulfonyl, ethanesulfonyl, isopropanesulfonyl, cyclopropanesulfonyl, p-toluenesulfonyl, hydroxyethyl, deuterated methanesulfonyl, deuterated ethanesulfonyl, deuterated isopropanesulfonyl, deuterated cyclopropanesulfonyl, deuterated formyl, and deuterated acetyl.

[0025] R 10 Selected from C 2-10 Heterocyclic group, C 6-15 Aromatic ring group, C 3-10 Heterocyclic aryl; the above-mentioned heterocyclic group, aryl group, heterocyclic aryl group may optionally be substituted by one or more hydrogen, halogen, hydroxyl, amino, nitro, methyl, ethyl, isopropyl, carboxyl, methoxy, ethoxy, isopropoxy, methylamino, ethylamino, isopropylamino, mercapto, mercaptomethyl, mercaptoethyl, acetyl, trifluoromethyl, methanesulfonyl, ethanesulfonyl;

[0026] R 11 Selected from C 2-10 Heterocyclic group, C 3-10 Heterocyclic aryl; the above-mentioned heterocyclic groups and heterocyclic aryl groups may optionally be substituted by one or more hydrogens, halogens, hydroxyl groups, amino groups, nitro groups, methyl groups, ethyl groups, isopropyl groups, carboxyl groups, methoxy groups, ethoxy groups, isopropoxy groups, methylamino groups, ethylamino groups, isopropylamino groups, mercapto groups, mercaptomethyl groups, mercaptoethyl groups, acetyl groups, trifluoromethyl groups, methanesulfonyl groups, and ethanesulfonyl groups;

[0027] G is selected from hydrogen, Hydroxyethyl;

[0028] When T is selected from hydrogen;

[0029] G is selected from Hydroxyethyl;

[0030] When T is selected from sodium or potassium; G is selected from hydrogen, and R9 is selected from deuterated methanesulfonyl, deuterated ethanesulfonyl, deuterated isopropanesulfonyl, deuterated cyclopropanesulfonyl, deuterated formyl, or deuterated acetyl.

[0031] T is selected from hour;

[0032] G is selected from hydrogen;

[0033] Furthermore, T and G are not both selected from hydrogen;

[0034] R a R b Each is independently selected from hydrogen and C. 1-6 Alkyl, or R a R b Connected to form a ring;

[0035] R c R d Each is independently selected from hydrogen and C. 1-6 Alkyl, or R c R d Connected to form a ring;

[0036] R 12 R 13 Each is independently selected from hydrogen, metal ions, and C. 1-6 Alkyl, C 3-6 carbon cyclo group, C 2-10 Heterocyclic group, C 3-10 Heterocyclic aryl, or R 12 R 13 Linked into a ring; the above-mentioned heterocyclic groups and heterocyclic aryl groups may optionally be substituted by one or more hydrogens, halogens, hydroxyl groups, amino groups, nitro groups, methyl groups, ethyl groups, isopropyl groups, carboxyl groups, methoxy groups, ethoxy groups, isopropoxy groups, methylamino groups, ethylamino groups, isopropylamino groups, mercapto groups, mercaptomethyl groups, mercaptoethyl groups, acetyl groups, trifluoromethyl groups, methanesulfonyl groups, and ethanesulfonyl groups;

[0037] R 14 Selected from C 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 carbon cyclo group, C 2-10 Heterocyclic group, C 3-10 Heterocyclic aryl, deuterated C 1-6 Alkyl; the above-mentioned alkyl, alkoxy, carbocyclic, heterocyclic, and heterocyclic aryl groups may optionally be substituted with one or more hydrogens, halogens, hydroxyl groups, amino groups, nitro groups, methyl groups, ethyl groups, isopropyl groups, carboxyl groups, methoxy groups, ethoxy groups, isopropoxy groups, methylamino groups, ethylamino groups, isopropylamino groups, mercapto groups, mercaptomethyl groups, mercaptoethyl groups, acetyl groups, trifluoromethyl groups, methanesulfonyl groups, and ethanesulfonyl groups.

[0038] The compound has the structure of Formula II as follows:

[0039] The substituents in Formula II are defined as defined in Formula I.

[0040] The compound described herein has the structure of Formula III as follows:

[0041] The substituents in Formula III are defined as defined in Formula I.

[0042] The compound has the following structure:

[0043] A pharmaceutical composition comprising the said compound or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

[0044] The use of the compound and / or the pharmaceutical composition described herein for the preparation of an anti-HIV agent.

[0045] A method of treating an individual infected with HIV includes administering a therapeutically effective amount of the compound and / or the pharmaceutical composition to the mammal requiring treatment.

[0046] In this invention, as selected for a more detailed understanding of the invention, the terms have the following definitions.

[0047] In this article, "amino" refers to a functional group having one nitrogen atom and 0 to 2 hydrogen atoms.

[0048] In this article, halogen refers to fluorine, chlorine, bromine, or iodine atoms.

[0049] The "C" in this article 1-6 "Alkyl" refers to a straight-chain, branched, or unbranched saturated aliphatic hydrocarbon group containing up to 6 carbon atoms. Specific alkyl examples are selected from methyl, ethyl, isopropyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 2,2-dimethylpropyl, etc.

[0050] The "C" in this article 1-6 "Alkoxy" refers to an alkyl group containing up to 6 carbon atoms with -O- or -OH groups inserted at any reasonable position. This group can be straight-chain, branched, or unbranched. Specific examples are selected from methoxy, ethoxy, isopropoxy, tert-butoxy, isopentoxy, cyclopentoxy, 2-methoxybutyl, 2-ethoxypropyl, 2-hydroxybutyl, etc.

[0051] The "C" in this article 1-6"Alkylamino" refers to an alkyl group containing up to 6 carbon atoms with -N-, -NH, or -NH2 groups inserted at any reasonable position. This group can be straight-chain, branched, or unbranched. Specific examples are selected from methylamino, ethylamino, isopropylamino, tert-butylamino, cyclopentanamino, 2-methylaminobutyl, 2-ethylaminopropyl, 2-aminobutyl, etc.

[0052] The "C" in this article 2-8 "Alkenyl" indicates a straight-chain, branched, or unbranched hydrocarbon group containing up to 8 carbon atoms and at least one carbon-carbon double bond. Specific examples are selected from vinyl, allyl, cis-2-pentenyl, 3-methyl-2-pentenyl, 2-methyl-2-pentenyl, etc.

[0053] The "C" in this article 2-8 "Alynyl" indicates that the molecule contains up to 8 carbon atoms and at least one carbon-carbon triple bond, and is a straight-chain, branched or unbranched hydrocarbon group. Specific examples are selected from ethynyl, propynyl, methylisopropylethynyl, prop-1-ynylcyclopropane, 1-pentynyl, 5-methyl-3-hexynyl, etc.

[0054] The "C" in this article 3-10 "Carbocyclic group" refers to a saturated or unsaturated aliphatic cyclic hydrocarbon group containing 3 to 10 carbon atoms in the molecule. Specific examples are selected from cyclopropyl, cyclobutyl, cyclopentyl, methylcyclopentyl, cyclohexyl, cyclohexenyl, cyclopentenyl, 1,3-cyclohexadienyl, etc.

[0055] The "C" in this article 2-8 "Heterocyclic group" refers to a saturated or unsaturated aliphatic cyclic group containing 1 to 8 carbon atoms and 1 to 6 heteroatoms in a molecule. It may contain one or more rings. The rings of such polycyclic heterocyclic alkyl groups may have different linkages, such as fused, bridged, spirocyclic, etc. Specific examples are selected from ethylene oxide, pyrrolidinyl, furanyl, piperidinyl, piperazinyl, pyrazinyl, pyranyl, tetrahydro-3-thiophenolyl, sulfide cyclopentyl, etc.

[0056] The "C" in this article 6-15 "Aryl" indicates a group containing 6 to 15 carbon atoms and at least one aromatic ring. Besides covalent groups, the rings of a polycyclic aryl group can have different linkages, such as fusion or bridging, and the fused rings can be saturated or unsaturated. Specific examples are selected from phenyl, naphthyl, diphenyl, α-tetrahydronaphthyl, indenyl, indenyl, benzopyrazine, 3,4-dihydro-1H-benzopyranyl, etc.

[0057] The "C" in this article 3-10"Heteroaryl" refers to an aromatic heterocyclic group containing 3 to 10 carbon atoms and 1 to 6 heteroatoms. Besides the covalent groups, the rings of the polycyclic aryl group can have different linkages, such as fusion or bridging, and the fused rings can be saturated or unsaturated. Specific examples are selected from thiophene, imidazolyl, pyrazolyl, isopyrazolyl, pyridinyl, pyrimidinyl, pyridazinyl, indolyl, quinolinyl, isoquinolinyl, quinoxalinyl, thiazolyl, purine, 5,6,7,8-tetrahydroquinolinyl, etc.

[0058] In this article, heteroatoms refer to oxygen atoms, nitrogen atoms, sulfur atoms, phosphorus atoms, boron atoms, selenium atoms, etc.

[0059] The compounds containing double bonds in this invention include all configurational isomers, such as cis and trans isomers.

[0060] The compounds of the present invention also contain their tautomers, such as positive ion shift isomers, negative ion shift isomers, bimolecular proton shift isomers, intracyclic tautomers, ring-chain tautomers, keto-enol tautomers, amide-imine isomers, lactam-lactam isomers, etc.

[0061] Compounds containing double bonds include all configurational isomers, such as cis and trans isomers.

[0062] The compounds of the present invention have asymmetric centers, and therefore the present invention relates to the use of optical isomers and stereoisomers of all such compounds and mixtures thereof.

[0063] The present invention also relates to the use of compounds having tautomers and mixtures thereof.

[0064] The compounds of this invention contain a basic nitrogen atom (heterocyclic or aliphatic amino group, etc.), which is readily oxidized by oxidizing agents such as oxygen in the air or hydrogen peroxide, selected from N-oxides, to generate other compounds of this invention. Therefore, the resulting N-oxide derivatives are selected as part of the compounds of this invention.

[0065] In this paper, "independently" means that when there are more than one variable, the choice of each instance of a substituent from the available variable definitions is independent of the other choices used to define the variables. Therefore, each substituent may be the same as or different from the other substituents.

[0066] In this invention, "individual" refers to warm-blooded animals, such as humans, rats, guinea pigs, mice, gerbils, rabbits, dogs, pigs, sheep, monkeys, chickens, ducks, geese, cats, cattle, horses, chimpanzees, etc.

[0067] In this article, "treatment" refers to actions taken to inhibit the progression of an applicable disorder or one or more conditions, or to reverse symptoms, and also includes adjunctive treatment for conditions.

[0068] The compounds of this invention can be used in the form of inorganic or organic acid derivatives selected from salts, depending on the characteristics of the compound, such as enhancing drug stability, water solubility, and achieving satisfactory solubility. They can also be used as adjuncts to separation, purification, and / or resolution. Salts formed with inorganic acids include hydrochloride, hydrobromide, sulfate, and phosphate; organic acid salts include formate, acetate, lactate, pyruvate, malate, maleate, malonate, fumarate, tartrate, citrate, aspartate, glutamate, benzoate, cinnamate, methanesulfonate, ethanesulfonate, p-toluenesulfonate, and salicylate; salts formed by combining with inorganic bases include sodium salts, potassium salts, ammonium salts, calcium salts, magnesium salts, zinc salts, and aluminum salts; and organic base salts include diethylamine salts, triethylamine salts, ethanolamine salts, aminobutanetriol salts, dicyclohexylamine salts, lysine salts, arginine salts, histidine salts, procaine salts, choline salts, betaine salts, glucosamine salts, methylglucosamine salts, piperazine salts, and piperidine salts.

[0069] The present invention provides a pharmaceutical composition comprising the compound, or isomers of the compound and its salts, and a pharmaceutically acceptable medium, diluent, or carrier.

[0070] In this article, "pharmaceutically acceptable mediators, diluents, and carriers" refers to diluents, starch, dextrin, sugars, mannitol, microcrystalline cellulose, oils, binders, humectants, distilled water, ethanol, starch pastes, disintegrants, lubricants, and flow aids, etc.

[0071] The compounds and compositions thereof provided by this invention can alleviate and / or treat cardiovascular and cerebrovascular diseases, organ pain, diabetes, inflammation and other related diseases.

[0072] The compounds and compositions provided by this invention can be prepared into pharmaceutical preparations for prevention or treatment of individuals requiring treatment through various administration methods such as oral, intranasal, oral, skin, and intravenous injection. Detailed Implementation

[0073] The present invention is further described below with reference to embodiments, but these embodiments are not intended to limit the scope of the invention. Unless otherwise specified, the experimental solvents or reagents used in the following embodiments were commercially available and used directly without further purification.

[0074] Example 1: Preparation and characterization of compound N-5

[0075] Compound N-2: Under nitrogen protection, 95 mL (1.0 M) of bis(trimethylsilylamine)lithium solution was slowly added to a THF solution of N-1 (12.1 g / 0.1 mol) and ethyl cyanoacetate (11.2 g / 0.1 mol) in 300 mL. The temperature was controlled below -10 °C during the dropwise addition. After the addition was complete, the reaction was continued for 4 hours in an ice bath. Petroleum ether (600 mL) was slowly added. The solids of the solution were filtered, and the filter cake was washed with petroleum ether (50 mL). The filter cake was then collected directly for the next step.

[0076] Compound N-4: N-3 (15.4 g / 0.1 mol) was added to an ethanol solution (300 mL) of compound N-2 obtained in the upward step under nitrogen protection, followed by ethanol hydrochloride (10 mL). The mixture was reacted at 60 °C for 6 hours, cooled to room temperature, concentrated, and extracted with ethyl acetate (200 mL × 2). The extract was washed with saturated brine (200 mL × 1), dried over anhydrous sodium sulfate, concentrated, and the residue was separated by column chromatography to give 5.6 g of compound N-4 (22% yield). MS: 256.3 [M+1].

[0077] Compound N-5: Potassium carbonate (2.7 g / 0.02 mol) was added to an ethanol solution (30 mL) of compound N-4 (2.5 g / 0.01 mol) obtained in the upward step under nitrogen protection, followed by the addition of water (2 mL). The mixture was reacted at 70 °C for 16 hours, cooled to room temperature, concentrated, and extracted with water (50 mL). The mixture was then washed with ethyl acetate (50 mL × 2), washed with saturated brine (50 mL × 1), dried over anhydrous sodium sulfate, concentrated, and the residue was separated by column chromatography to yield 1.4 g of compound N-5 (62% yield). MS: 228.0 [M+1].

[0078] Example 2: Preparation and characterization of compound N-10

[0079] Compound N-7: Under nitrogen protection, silver nitrate (33.8 g / 0.2 mol) and triethylamine (20.2 g / 0.2 mol) were added to a THF (500 mL) solution of N-6 (22.3 g / 0.1 mol) and iodine (25.3 g / 0.1 mol). The reaction was carried out at room temperature for 15 hours. The mixture was filtered, the filtrate was concentrated, and 0.01 M sodium bicarbonate solution (300 mL) was added. The mixture was extracted with ethyl acetate (300 mL × 2), washed with saturated brine (300 mL × 1), dried over anhydrous sodium sulfate, concentrated, and the residue was separated by column chromatography to give 11.0 g of compound N-7 (40% yield). MS: 275.1 [M+1].

[0080] Compound N-8: Iron powder (5.6 g / 0.1 mol) and ammonium chloride (5.4 g / 0.1 mol) were added to a methanol solution (200 mL) of N-7 (13.7 g / 0.05 mol). The mixture was heated to reflux for 16 hours, cooled to room temperature, filtered, concentrated, and the residue was extracted with 0.01 M sodium bicarbonate solution (100 mL). The residue was extracted with ethyl acetate (100 mL × 2), washed with saturated brine (100 mL × 1), dried over anhydrous sodium sulfate, concentrated, and the residue was separated by column chromatography to give 5.6 g of compound N-8 (45% yield). MS: 245.2 [M+1].

[0081] Compound N-9: Under nitrogen protection, cyclopropanesulfonyl chloride (1.4 g / 0.01 mol) and triethylamine (2.0 g / 0.02 mol) were added to a dichloromethane solution (50 mL) of N-8 (2.4 g / 0.01 mol). The mixture was stirred at room temperature for 3 hours, filtered, washed with water (30 mL × 1), washed with saturated brine (30 mL × 1), dried over anhydrous sodium sulfate on the organic phase, concentrated, and the residue was separated by column chromatography to give 1.4 g of compound N-9 (40% yield). MS: 349.1 [M+1].

[0082] Compound N-10: Under nitrogen protection, 60% sodium hydroxide (0.8 g / 0.02 mol) was added to a THF solution (50 mL) of N-9 (3.5 g / 0.01 mol), and the mixture was stirred at room temperature for 1 hour. Chloroacetonitrile (0.8 g / 0.01 mol) was slowly added, with the internal temperature controlled not to exceed 5 °C. After the addition was complete, the reaction was continued at room temperature for 4 hours. Ammonium chloride solution (0.01 M / 10 mL) was slowly added, and the mixture was concentrated. Water (30 mL) was added, and the mixture was extracted with ethyl acetate (50 mL × 2). The mixture was washed with saturated brine (50 mL × 1), dried over anhydrous sodium sulfate, and concentrated. The residue was separated by column chromatography to give 1.6 g of compound N-10 (42% yield). MS: 388.1 [M+1].

[0083] Example 3: Preparation and characterization of compound N-15

[0084] Compound N-12: Under nitrogen protection, (S)-2-methylpropane-2-sulfonamide (1.2 g / 0.01 mol) and tetraethyl titanate (4.6 g / 0.02 mol) were added to a 100 mL solution of N-11 (2.6 g / 0.01 mol) in toluene. After the addition was complete, the mixture was heated to 70 °C and reacted for 4 hours. The mixture was then cooled to room temperature, and 30 mL of 1.0 M hydrochloric acid was added. The mixture was stirred for 0.5 hours, and the aqueous phase was separated. The organic phase was dried over anhydrous sodium sulfate and concentrated. Ethyl acetate (15 mL) was added to the residue, and n-hexane (45 mL) was slowly added dropwise while stirring. A solid precipitated, which was filtered and dried to give 0.9 g of compound N-12 (25% yield). MS: 368.0 [M+1]. This was used directly in the next step.

[0085] Compound N-13: Under nitrogen protection, a borane dimethyl sulfide complex (10 mL / 2.0 M) was added to an N-12 (3.7 g / 0.01 mol) solution in 80 mL of THF. The temperature was controlled to not exceed -10 °C. After the addition was complete, the reaction was carried out in an ice bath for 5 hours. Then, 50 mL of 0.01 M sodium bicarbonate solution was added, and the mixture was stirred in an ice bath for 1 hour. The aqueous phase was separated, and 20 mL of 1.0 M ethyl hydrochloride solution was slowly added to the organic phase in an ice bath. The mixture was stirred for another 2 hours, and a solid precipitated. The solid was filtered and dried to give 0.5 g of compound N-13 (12% yield). MS: 391.4 [M+1]. This was used directly for the next step.

[0086] Compound N-14: Triethylamine (3.0 g / 0.03 mol) and di-tert-butyl dicarbonate (2.2 g / 0.01 mol) were added to a solution of N-13 (4.3 g / 0.01 mol) in 100 mL of dichloromethane under nitrogen protection. After the addition was complete, the mixture was reacted at room temperature for 6 hours. The mixture was washed with water (50 mL × 1) and saturated brine (50 mL × 1). The organic phase was concentrated, and the residue was separated by column chromatography to give 3.8 g of compound N-14 (77% yield). MS: 491.3 [M+1].

[0087] Compound N-15: Under nitrogen protection, tetrakis(triphenylphosphine)palladium (1.1 g / 0.1 mmol) and pinacol diborate (2.5 g / 0.01 mol) were added to a 1,4-dioxane (100 mL) solution of N-14 (4.9 g / 0.01 mol), followed by potassium acetate (2.0 g / 0.02 mol). After the addition was complete, the mixture was refluxed for 3 hours, cooled to room temperature, concentrated, and extracted with ethyl acetate (100 mL × 2). The extract was washed with water (100 mL × 1) and saturated brine (100 mL × 1). The organic phase was concentrated, and the residue was separated by column chromatography to give 1.2 g of compound N-15 (21% yield). MS: 539.2 [M+1].

[0088] Example 4: Preparation and characterization of compound N-18

[0089] Compound N-16: Under nitrogen protection, tetrakis(triphenylphosphine)palladium (1.1 g / 0.1 mmol) and N-10 (3.9 g / 0.01 mol) were added to a 1,4-dioxane (100 mL) solution of N-15 (5.4 g / 0.01 mol), followed by potassium acetate (2.0 g / 0.02 mol) and water (10 mL). The mixture was then refluxed for 4 hours, cooled to room temperature, concentrated, and extracted with ethyl acetate (100 mL × 2). The extract was washed with water (100 mL × 1) and saturated brine (100 mL × 1). The organic phase was concentrated, and the residue was separated by column chromatography to give 5.0 g of compound N-16 (66% yield). MS: 752.0 [M+1].

[0090] Compound N-17: Add HOBt (1.4 g / 0.01 mol) and EDCI (1.9 g / 0.01 mol) to a DCM (100 mL) solution of N-5 (2.3 g / 0.01 mol), then add triethylamine (5.1 g / 0.05 mol), and continue stirring for 1 hour.

[0091] Trifluoroacetic acid (2.3 g / 0.02 mol) was added to a DCM (50 mL) solution of N-16 (7.5 g / 0.01 mol), and the mixture was stirred at room temperature for 2 hours. The mixture was then slowly added to the solution, and the reaction was continued at room temperature for 16 hours. Ice water (100 mL) was added, and the mixture was extracted. The organic phase was separated, concentrated, and the residue was separated by column chromatography to give 4.7 g of compound N-17 (55% yield). MS: 861.1 [M+1].

[0092] Compound N-18: Under nitrogen protection, palladium dichloride bis(triphenylphosphine) dichloride (0.7 g / 1.0 mmol) and cuprous iodide (1.9 g / 0.01 mol) were added to a DMSO (50 mL) solution of N-17 (8.6 g / 0.01 mol), followed by triethylamine (2.0 g / 0.02 mol). After the addition was complete, the mixture was heated to 80 °C and reacted for 5 hours. The mixture was then cooled to room temperature, and water (150 mL) was added. The mixture was extracted with ethyl acetate (100 mL × 2), and the organic phase was washed with water (100 mL × 1) and saturated brine (100 mL × 1). The organic phase was concentrated, and the residue was separated by column chromatography to give 3.0 g of compound N-18 (33% yield). MS: 937.3 [M+1].

[0093] Example 5: Preparation and characterization of compound LN175-01

[0094] Compound N-19: Under nitrogen protection, 1.0 M hydrochloric acid (15 mL) was added to a solution of N-18 (9.4 g / 0.01 mol) in dichloromethane (150 mL). After the addition was complete, the mixture was stirred at room temperature for 2 hours. Under ice bath conditions, 0.5 M sodium bicarbonate solution (100 mL) was added, and the mixture was stirred for 0.5 hours. The organic phase was separated, washed with water (100 mL × 1), and washed with saturated brine (100 mL × 1). The organic phase was concentrated, and the residue was separated by column chromatography to give 2.9 g of compound N-19 (34% yield). MS: 865.1 [M+1].

[0095] Compound LN175-01: Under nitrogen protection, paraformaldehyde (0.20 g) and potassium carbonate (0.27 g / 2.0 mmol) were added to an anhydrous THF (20 mL) solution of N-19 (0.86 g / 1.0 mmol). The mixture was heated to reflux for 4 hours, cooled to room temperature, filtered, and the filtrate was slowly added to triethylamine (0.40 g / 4.0 mmol) and phosphorus oxychloride (0.15 g / 1.0 mmol) under nitrogen protection. The reaction was continued at room temperature for 16 hours. 0.01 M sodium bicarbonate solution (5 mL) was slowly added under ice bath, and the mixture was concentrated. The residue was directly separated by preparative liquid chromatography to give 0.16 g of compound LN175-01 (17% yield). MS: 975.1 [M+1]. 1H NMR(DMSO-d6)δ9.19-9.17(m,1H),8.29-8.17(m,2H),7.43-7.38(m,4H),7.19-7.10(m,2H),5.82(s,2H),5.25-5.23(m,1H),4.91 -4.63(m,4H),3.23-3.21(m,1H),3.01-2.95(m,3H),2.27-2.25(m,2H),1.62-1.51(m,6H),1.35-1.30(m,2H),1.08-1.04(m,2H).

[0096] Example 6: Preparation and characterization of compound LN175-02

[0097] Compound LN175-02: Under nitrogen protection, thionyl chloride (0.36 g / 2.0 mmol) was added to a 20 mL THF solution of LN175-01 (0.97 g / 1.0 mmol). After the addition was complete, the mixture was stirred for 2 hours. Then, methanol (1 mL) and triethylamine (0.30 g / 3.0 mmol) were added, and the reaction was stirred for another 5 hours. The mixture was concentrated, and the residue was directly separated by preparative liquid chromatography to give 0.13 g of compound LN175-02 (13% yield). MS: 1003.4 [M+1]. 1 H NMR(DMSO-d6)δ9.21-9.20(m,1H),8.30-8.18(m,2H),7.46-7.41(m,4H),7.20-7.12(m,2H),5.84(s,2H),5.26-5.24(m,1H),4.93-4.65(m,4H) ,3.74(s,3H),3.72(s,3H),3.25-3.23(m,1H),3.04-2.97(m,3H),2.28 -2.26(m,2H),1.65-1.53(m,6H),1.36-1.32(m,2H),1.09-1.05(m,2H).

[0098] Example 7: Preparation and characterization of compound LN175-03

[0099] Compound LN175-03: Under nitrogen protection, water (2 mL) and sodium hydroxide (0.4 g / 10.0 mmol) were added to a 20 mL solution of LN175-01 (0.97 g / 1.0 mmol). After the addition was complete, the mixture was stirred for 5 hours, filtered, and the resulting solid was recrystallized from acetone / water (3:1) to give 0.09 g of compound LN175-03 (9% yield). MS: 1041.2 [M+23]. 1H NMR(DMSO-d6)δ9.17-9.16(m,1H),8.26-8.18(m,2H),7.41-7.37(m,4H),7.16-7.09(m,2H),5.78(s,2H),5.23-5.21(m,1H),4.89 -4.61(m,4H),3.22-3.20(m,1H),3.00-2.95(m,3H),2.26-2.21(m,2H),1.60-1.50(m,6H),1.32-1.28(m,2H),1.05-1.01(m,2H).

[0100] Example 8: Preparation and characterization of compound LN175-05

[0101] Compound LN175-05: Under nitrogen protection, thionyl chloride (0.36 g / 2.0 mmol) was added to a 20 mL THF solution of LN175-01 (0.97 g / 1.0 mmol). After the addition was complete, the mixture was stirred for 2 hours. Then, ethylene glycol (1 mL) and triethylamine (0.30 g / 3.0 mmol) were added, and the reaction was stirred for another 5 hours. The mixture was concentrated, and the residue was directly separated by preparative liquid chromatography to give 0.12 g of compound LN175-05 (12% yield). MS: 100 1.4 [M+1]. 1 H NMR(DMSO-d6)δ9.24-9.23(m,1H),8.34-8.23(m,2H),7.43-7.40(m,4H),7.22-7.11(m,2H),5.81(s,2H),5.23-5.21(m,1H),4.88-4.63(m, 4H),4.39-4.34(m,4H),3.23-3.21(m,1H),3.02-2.95(m,3H),2.24-2 .21(m,2H),1.63-1.52(m,6H),1.33-1.30(m,2H),1.06-1.03(m,2H).

[0102] Example 9: Preparation and characterization of compound LN175-07

[0103] Compound N-20: Under nitrogen protection, paraformaldehyde (0.4 g) and triethylamine (0.2 g / 2.0 mmol) were added to an acetonitrile (10 mL) solution of N-19 (0.86 g / 1.0 mmol). After the addition was complete, the mixture was heated to reflux for 3 hours, cooled to room temperature, and concentrated under reduced pressure. The resulting residue N-20 was used directly in the next step.

[0104] Compound LN175-07: Acetyl chloride (0.08 g / 1.0 mmol) and triethylamine (0.2 g / 2.0 mmol) were added to an acetonitrile (20 mL) solution at N-20 under nitrogen protection. After the addition was complete, the mixture was reacted at room temperature for 4 hours, cooled to room temperature, concentrated under reduced pressure, and directly separated by preparative liquid chromatography to obtain 0.10 g of compound LN175-07 (11% yield). MS: 937.3 [M+1]. 1 H NMR(DMSO-d6)δ9.20-9.19(m,1H),8.24-8.20(m,2H),7.42-7.38(m,4H),7.14-7.10(m,2H),5.72(s,2H),5.22-5.20(m,1H),4.91 -4.64(m,4H),3.21-3.20(m,1H),3.01-2.96(m,3H),2.25-2.21(m,5H),1.62-1.52(m,6H),1.33-1.29(m,2H),1.04-1.01(m,2H).

[0105] Example 10: Preparation and characterization of compound N-26

[0106] Compound N-22: Under nitrogen protection, methanesulfonyl chloride (13.7 g / 0.12 mol) and triethylamine (20.0 g / 0.2 mol) were added to tetrahydrofuran (750 mL) containing N-21 (32.8 g / 0.1 mol). The mixture was heated to 60 °C and stirred for 5 hours. After filtration and concentration, dichloromethane (500 mL) was added, followed by washing with water (300 mL × 1) and saturated brine (300 mL × 1). The organic phase was dried over anhydrous sodium sulfate, concentrated, and the residue was separated by column chromatography to give 31.3 g of compound N-22 (77% yield). MS: 406.2 [M+1].

[0107] Compound N-23: Under nitrogen protection, tetrakis(triphenylphosphine)palladium (4.0 g / 3.5 mmol) and N-22 (30.0 g / 73.8 mmol) were added to a solution of N-15 (39.8 g / 73.8 mmol) in 1,4-dioxane (1000 mL). Potassium acetate (9.8 g / 0.1 mol) was added, followed by water (100 mL). The mixture was then refluxed for 6 hours, cooled to room temperature, concentrated, and extracted with ethyl acetate (1000 mL × 2). The extract was washed with water (1000 mL × 1) and saturated brine (1000 mL × 1). The organic phase was concentrated, and the residue was separated by column chromatography to yield 31.1 g of compound N-23 (57% yield). MS: 738.0 [M+1].

[0108] Compound N-24: Under nitrogen protection, palladium dichloride bis(triphenylphosphine) dichloride (3.0 g / 4.3 mmol) and cuprous iodide (1.9 g / 0.01 mol) were added to a DMSO (500 mL) solution of N-23 (30.5 g / 41.3 mmol). Triethylamine (5.1 g / 50.0 mmol) was then added. After the addition was complete, the mixture was heated to 90 °C and reacted for 16 hours. The mixture was then cooled to room temperature, and dichloromethane (500 mL) and trifluoroacetic acid (100 mL) were added. The mixture was stirred at room temperature for 2 hours. Triethylamine (200 mL) was added under ice bath conditions. The mixture was concentrated and extracted with dichloromethane (500 mL × 2). The organic phase was washed with water (500 mL × 1) and saturated brine (500 mL × 1). The organic phase was concentrated, and the residue was separated by column chromatography to give 11.9 g of compound N-24 (41% yield). MS: 704.2 [M+1].

[0109] Compound N-26: HOBt (1.4 g / 0.01 mol) and EDCI (1.9 g / 0.01 mol) were added to a DCM (100 mL) solution of N-25 (2.8 g / 0.01 mol), followed by the addition of triethylamine (2.0 g / 0.02 mol). The mixture was stirred for 1 hour after the reaction was complete. N-24 (7.0 g / 0.01 mol) was then added to the above reaction solution, and the reaction was continued at room temperature for 15 hours. Ice water (100 mL) was added, and the mixture was extracted. The organic phase was separated, concentrated, and the residue was separated by column chromatography to obtain 3.4 g of compound N-26 (35% yield). MS: 968.3 [M+1].

[0110] Example 11: Preparation and characterization of compound LN175-25

[0111] Compound LN175-25: Under nitrogen protection, paraformaldehyde (0.20 g) and potassium carbonate (0.27 g / 2.0 mmol) were added to an anhydrous THF (20 mL) solution of N-26 (0.97 g / 1.0 mmol). The mixture was heated to reflux for 4 hours, cooled to room temperature, filtered, and the filtrate was slowly added to triethylamine (0.40 g / 4.0 mmol) and phosphorus oxychloride (0.15 g / 1.0 mmol) under nitrogen protection. The reaction was continued at room temperature for 16 hours. 0.01 M sodium bicarbonate solution (5 mL) was slowly added under ice bath, and the mixture was concentrated. The residue was directly separated by preparative liquid chromatography to give 0.15 g of compound LN175-25 (14% yield). MS: 1078.1 [M+1]. 1HNMR(DMSO-d6)δ9.20-9.18(m,1H),7.80-7.78(m,2H),7.44-7.42(m,1H), 7.04-7.01(m,1H),6.52-6.50(m,2H),5.73(s,2H),4.90-4.68(m,4H),4.0 7-4.05(m,1H),3.52-3.43(m,4H),2.97-2.94(m,3H),2.67-2.41(m,3H),2 .34-2.33(m,1H),1.72-1.64(m,6H),1.42-1.39(m,1H),1.05-1.03(m,1H).

[0112] Example 12: Preparation and characterization of compound LN175-26

[0113] Compound LN175-26: Under nitrogen protection, thionyl chloride (0.04 g / 0.2 mmol) was added to a 20 mL THF solution of LN175-25 (0.11 g / 0.1 mmol). After the addition was complete, the mixture was stirred for 2 hours. Then, methanol (1 mL) and triethylamine (0.30 g / 3.0 mmol) were added, and the reaction was stirred for another 6 hours. The mixture was concentrated, and the residue was directly separated by preparative liquid chromatography to give 10.0 mg of compound LN175-26 (9% yield). MS: 1106.3 [M+1]. 1 H NMR(DMSO-d6)δ9.20-9.18(m,1H),7.80-7.78(m,2H),7.44-7.42(m,1H),7 .04-7.01(m,1H),6.52-6.50(m,2H),5.73(s,2H),4.90-4.68(m,4H),4.07 -4.05(m,1H),3.52-3.43(m,4H),2.97-2.94(m,3H),2.67-2.41(m,3H),2. 34-2.33(m,1H),1.72-1.64(m,6H),1.42-1.39(m,1H),1.05-1.03(m,1H).

[0114] Example 13: Preparation and characterization of compound LN175-27

[0115] Compound LN175-27: Under nitrogen protection, 2 mL of water and 0.04 g of sodium hydroxide (1.0 mmol) were added to a 20 mL solution of LN175-25 (0.11 g / 0.1 mmol). After the addition was complete, the mixture was stirred for 15 hours, filtered, and the resulting solid was recrystallized from acetone / water (4:1) to give 9.0 mg of compound LN175-27 (8% yield). MS: 1144.4 [M+23]. 1 H NMR(DMSO-d6)δ9.17-9.16(m,1H),7.79-7.76(m,2H),7.41-7.38(m,1H),7 .02-7.00(m,1H),6.50-6.47(m,2H),5.70(s,2H),4.87-4.85(m,4H),4.07 -4.05(m,1H),3.51-3.42(m,4H),2.96-2.94(m,3H),2.68-2.44(m,3H),2. 33-2.31(m,1H),1.71-1.65(m,6H),1.43-1.40(m,1H),1.04-1.03(m,1H).

[0116] Example 14: Preparation and characterization of compound LN175-32

[0117] Compound LN175-32: Under nitrogen protection, paraformaldehyde (0.20 g) and potassium carbonate (0.27 g / 2.0 mmol) were added to anhydrous THF (20 mL) solution of N-26 (0.97 g / 1.0 mmol). The mixture was heated to reflux for 4 hours, cooled to room temperature, filtered, and the filtrate was slowly added to triethylamine (0.40 g / 4.0 mmol) and isobutyryl chloride (0.11 g / 1.0 mmol) under nitrogen protection. The reaction was continued at room temperature for 21 hours. 0.01 M sodium bicarbonate solution (5 mL) was slowly added under ice bath, and the mixture was concentrated. The residue was directly separated by preparative liquid chromatography to give 0.37 g of compound LN175-32 (35% yield). MS: 1068.1 [M+1]. 1H NMR(DMSO-d6)δ9.19-9.18(m,1H),7.81-7.79(m,2H),7.43-7.40(m,1H),7 .04-7.03(m,1H),6.55-6.52(m,2H),5.75(s,2H),4.83-4.80(m,4H),4.09 -4.07(m,1H),3.53-3.45(m,4H),2.93-2.90(m,3H),2.66-2.47(m,3H),2. 31-2.28(m,1H),1.70-1.65(m,6H),1.42-1.39(m,1H),1.13-1.02(m,7H).

[0118] Example 15: Preparation and characterization of compound LN175-34

[0119] Compound LN175-34: Under nitrogen protection, paraformaldehyde (0.20 g) and potassium carbonate (0.27 g / 2.0 mmol) were added to anhydrous THF (20 mL) solution of N-26 (0.97 g / 1.0 mmol). The mixture was heated to reflux for 4 hours, cooled to room temperature, filtered, and the filtrate was slowly added to triethylamine (0.40 g / 4.0 mmol) and methyl chloroformate (0.09 g / 1.0 mmol) under nitrogen protection. The reaction was continued at room temperature for 16 hours. 0.01 M sodium bicarbonate solution (5 mL) was slowly added under ice bath, and the mixture was concentrated. The residue was directly separated by preparative liquid chromatography to give 0.31 g of compound LN175-34 (30% yield). MS: 1056.1 [M+1]. 1 H NMR(DMSO-d6)δ9.21-9.20(m,1H),7.82-7.80(m,2H),7.43-7.42(m,1H),7.05-7 .03(m,1H),6.53-6.49(m,2H),6.05(s,2H),5.74(s,2H),4.90-4.86(m,4H),4.09 -4.06(m,1H),3.74(s,3H),3.53-3.44(m,4H),2.97-2.94(m,3H),2.70-2.46(m, 3H),2.36-2.32(m,1H),1.76-1.68(m,6H),1.44-1.40(m,1H),1.05-1.04(m,1H).

[0120] Example 16: Preparation and characterization of compound LN175-37

[0121] Compound LN175-37:

[0122] Under nitrogen protection, 60% sodium hydroxide (0.08 g / 2.0 mmol) was added to an anhydrous THF (20 mL) solution of N-19 (0.86 g / 1.0 mmol). After stirring at room temperature for 1 hour, di-tert-butylchloromethyl phosphate (0.26 g / 1.0 mmol) was added. After the addition was complete, the mixture was stirred at room temperature for 2 hours. Then, di-tert-butylchloromethyl phosphate (0.26 g / 1.0 mmol) was added again. After the addition was complete, the mixture was stirred at room temperature for 16 hours. The mixture was filtered, and trifluoroacetic acid (2 mL) was added to the filtrate. The mixture was stirred for another 6 hours. The solution was adjusted to alkalinity with triethylamine, concentrated, and the residue was purified by pre-HPLC to give 0.08 g of compound LN175-37 (8% yield). MS: 975.3 [M+1]. 1 H NMR(DMSO-d6)δ9.18-9.17(m,1H),8.22-8.20(m,2H),7.43-7.39(m,4H),7.13-7.11(m,2H),5.73(s,2H),5.23-5.21(m,1H),4.93 -4.68(m,4H),3.22-3.20(m,1H),3.03-2.97(m,3H),2.26-2.20(m,5H),1.63-1.54(m,6H),1.35-1.31(m,2H),1.06-1.03(m,2H).

[0123] Example 17: Preparation and characterization of compound LN175-38

[0124] Compound LN175-38:

[0125] Under nitrogen protection, thionyl chloride (0.36 g / 2.0 mmol) was added to a 20 mL THF solution of LN175-37 (0.97 g / 1.0 mmol). After the addition was complete, the mixture was stirred for 1 hour. Then, methanol (1 mL) and triethylamine (0.30 g / 3.0 mmol) were added, and the reaction was stirred for another 6 hours. The mixture was concentrated, and the residue was directly separated by preparative liquid chromatography to give 0.11 g of compound LN175-38 (11% yield). MS: 100 3.0 [M+1]. 1H NMR(DMSO-d6)δ9.16-9.15(m,1H),8.27-8.25(m,2H),7.41-7.38(m,4H),7.17-7.11(m,2H),5.88(s,2H),5.24-5.22(m,1H),4.90-4.64(m,4H) ,3.79(s,3H),3.77(s,3H),3.25-3.22(m,1H),3.00-2.94(m,3H),2.26 -2.24(m,2H),1.60-1.52(m,6H),1.34-1.31(m,2H),1.09-1.06(m,2H).

[0126] Example 18: Preparation and characterization of compound LN175-40

[0127] Compound LN175-40: Under nitrogen protection, 60% sodium hydride (0.08 g / 2.0 mmol) was added to an anhydrous THF (20 mL) solution of N-19 (0.86 g / 1.0 mmol). After stirring at room temperature for 1 hour, dimethyl chloromethyl carbonate (0.12 g / 1.0 mmol) was added. After the addition was complete, the mixture was stirred at room temperature for 15 hours. The mixture was filtered, and 0.01 M ammonium chloride solution (1 mL) was added to the filtrate. The mixture was stirred for another hour, filtered, concentrated, and the residue was purified by pre-HPLC to give 0.11 g of compound LN175-40 (11% yield). MS: 953.3 [M+1]. 1 H NMR(DMSO-d6)δ9.17-9.16(m,1H),8.22-8.20(m,2H),7.43-7.40(m,4H),7.16-7.12(m,2H),5.85(s,2H),5.23-5.21(m,1H),4.91-4.67 (m,4H),3.76(s,3H),3.23-3.20(m,1H),3.01-2.95(m,3H),2.23-2.20(m,2H),1.61-1.53(m,6H),1.33-1.31(m,2H),1.08-1.05(m,2H).

[0128] Example 19: Preparation and Characterization of Compound LN175-43

[0129] Compound LN175-43: Under nitrogen protection, 60% sodium hydroxide (0.08 g / 2.0 mmol) was added to an anhydrous THF (20 mL) solution of N-26 (0.97 g / 1.0 mmol). After stirring at room temperature for 1 hour, di-tert-butylchloromethyl phosphate (0.26 g / 1.0 mmol) was added. After the addition was complete, the mixture was stirred at room temperature for 2 hours. Then, di-tert-butylchloromethyl phosphate (0.26 g / 1.0 mmol) was added. After the addition was complete, the mixture was stirred at room temperature for 15 hours. The mixture was filtered, and trifluoroacetic acid (2 mL) was added to the filtrate. The mixture was stirred for another 6 hours. The solution was adjusted to alkalinity with triethylamine, concentrated, and the residue was purified by pre-HPLC to give 0.16 g of compound LN175-43 (15% yield). MS: 1078.4 [M+1]. 1 H NMR(DMSO-d6)δ9.15-9.14(m,1H),7.75-7.74(m,2H),7.38-7.36(m,1H),7 .01-7.00(m,1H),6.48-6.45(m,2H),5.73(s,2H),4.85-4.81(m,4H),4.05 -4.01(m,1H),3.50-3.43(m,4H),2.93-2.90(m,3H),2.66-2.45(m,3H),2. 32-2.30(m,1H),1.68-1.62(m,6H),1.40-1.38(m,1H),1.02-1.00(m,1H).

[0130] Example 20: Preparation and characterization of compound LN175-44

[0131] Compound LN175-44: Under nitrogen protection, thionyl chloride (0.04 g / 0.2 mmol) was added to a 20 mL THF solution of LN175-43 (0.11 g / 0.1 mmol). After the addition was complete, the mixture was stirred for 2 hours. Then, methanol (1 mL) and triethylamine (0.30 g / 3.0 mmol) were added, and the reaction was stirred for another 5 hours. The mixture was concentrated, and the residue was directly separated by preparative liquid chromatography to give 14.4 mg of compound LN175-44 (13% yield). MS: 1106.3 [M+1]. 1H NMR(DMSO-d6)δ9.16-9.15(m,1H),7.77-7.75(m,2H),7.40-7.38(m,1H),7.04-7 .03(m,1H),6.49-6.47(m,2H),5.77(s,2H),4.86-4.83(m,4H),4.06-4.03(m,1H) ,3.77(s,3H),3.75(s,3H),3.53-3.46(m,4H),2.95-2.92(m,3H),2.61-2.42(m, 3H),2.30-2.29(m,1H),1.67-1.62(m,6H),1.41-1.37(m,1H),1.03-1.01(m,1H).

[0132] Example 21: Preparation and characterization of compound LN175-45

[0133] Compound LN175-45: Under nitrogen protection, 2 mL of water and 0.04 g of sodium hydroxide (1.0 mmol) were added to a 20 mL solution of LN175-43 (0.11 g / 0.1 mmol). After the addition was complete, the mixture was stirred for 20 hours, filtered, and the resulting solid was recrystallized from acetone / water (3:1) to give 19.1 mg of compound LN175-45 (17% yield). MS: 1144.1 [M+23]. 1 H NMR(DMSO-d6)δ9.16-9.15(m,1H),7.76-7.74(m,2H),7.39-7.37(m,1H),7 .02-7.01(m,1H),6.50-6.46(m,2H),5.74(s,2H),4.86-4.82(m,4H),4.07 -4.04(m,1H),3.56-3.47(m,4H),2.95-2.91(m,3H),2.63-2.46(m,3H),2. 33-2.31(m,1H),1.69-1.63(m,6H),1.41-1.38(m,1H),1.04-1.02(m,1H).

[0134] Example 22: Preparation and characterization of compound LN175-46

[0135] Compound LN175-46: Under nitrogen protection, 60% sodium hydroxide (0.08 g / 2.0 mmol) was added to an anhydrous THF (20 mL) solution of N-26 (0.97 g / 1.0 mmol). After stirring at room temperature for 1 hour, dimethyl chloromethyl carbonate (0.13 g / 1.0 mmol) was added. After the addition was complete, the mixture was stirred at room temperature for 20 hours. Ammonium chloride solution (0.01 M / 2.0 mL) was added, filtered, concentrated, and the residue was purified by pre-HPLC to give 0.12 g of compound LN175-46 (12% yield). MS: 1056.1 [M+1]. 1 H NMR(DMSO-d6)δ9.26-9.24(m,1H),7.83-7.79(m,2H),7.45-7.43(m,1H),7.0 5-7.03(m,1H),6.53-6.51(m,2H),5.75(s,2H),4.93-4.62(m,4H),4.10-4.08 (m,1H),3.54-3.44(m,4H),3.37(s,3H),3.00-2.98(m,3H),2.70-2.42(m,3H) ,2.35-2.31(m,1H),1.76-1.69(m,6H),1.43-1.41(m,1H),1.05-1.03(m,1H).

[0136] Example 23: Preparation and characterization of compound LN175-48

[0137] Compound LN175-48: Under nitrogen protection, 60% sodium hydroxide (0.08 g / 2.0 mmol) was added to an anhydrous THF (20 mL) solution of N-26 (0.97 g / 1.0 mmol). After stirring at room temperature for 1 hour, chloromethyl isobutyrate (0.14 g / 1.0 mmol) was added. After the addition was complete, the mixture was stirred at room temperature for 18 hours. Ammonium chloride solution (0.01 M / 2.0 mL) was added, filtered, concentrated, and the residue was purified by pre-HPLC to give 0.19 g of compound LN175-48 (18% yield). MS: 1068.1 [M+1]. 9.25(s,1H),7.83-7.79(m,1H),7.44-7.43(m,1H),7.05-7.03(m,1H),6.52-6.5 0(m,3H),5.76(s,2H),4.93(s,1H),4.89(s,2H),4.78(s,2H),4.08-4.05(m,2H) ,3.54(s,3H),2.99-2.97(m,1H),2.70(s,3H),2.53-2.50(m,1H),2.35-2.33(m, 1H),1.76-1.63(m,6H),1.42-1.39(m,1H),1.17-1.11(m,6H),1.00-0.98(m,1H).

[0138] Example 24: Preparation and characterization of compound LN175-55

[0139] Compound LN175-55: Under nitrogen protection, phosphorus oxychloride (0.15 g / 1.0 mmol) and potassium carbonate (0.41 g / 3.0 mmol) were added to a DMF (10 mL) solution containing N-26 (0.97 g / 1.0 mmol). The internal temperature was maintained at 0-5 °C during the addition process. After the addition was complete, the mixture was stirred at room temperature for 5 hours. The mixture was filtered, and ice water (30 mL) was added. The mixture was extracted with dichloromethane (30 mL × 3), and the organic phase was washed with water (30 mL). The residue obtained after concentration was purified by pre-HPLC to give 0.16 g of compound LN175-55 (15% yield). MS: 1048.3 [M+1]. 1H NMR(DMSO-d6)δ9.14-9.13(m,1H),7.72-7.70(m,2H),7.36-7.35(m,1H),7.02-7.01(m,1H),6.44-6.42(m,2H),4.82-4.79(m,4H),4.04-4.02 (m,1H),3.51-3.44(m,4H),2.93-2.90(m,3H),2.59-2.43(m,3H),2.31 -2.29(m,1H),1.63-1.54(m,6H),1.38-1.35(m,1H),1.02-1.00(m,1H).

[0140] Example 25: Preparation and characterization of compound LN175-56

[0141] Compound LN175-56: Under nitrogen protection, 2 mL of water and 0.08 g of sodium hydroxide (2.0 mmol) were added to a 20 mL solution of LN175-55 (0.21 g / 0.2 mmol). After the addition was complete, the mixture was stirred for 15 hours, filtered, and the resulting solid was recrystallized from acetone / water (5:1) to give 30.6 mg of compound LN175-56 (14% yield). MS: 1114.1 [M+23]. 1 H NMR(DMSO-d6)δ9.13-9.11(m,1H),7.71-7.70(m,2H),7.34-7.33(m,1H),7.01-7.00(m,1H),6.42-6.39(m,2H),4.81-4.77(m,4H),4.02-4.01 (m,1H),3.49-3.43(m,4H),2.91-2.88(m,3H),2.56-2.42(m,3H),2.30 -2.28(m,1H),1.62-1.52(m,6H),1.36-1.33(m,1H),1.01-0.99(m,1H).

[0142] Example 26: Preparation and characterization of compound LN175-57

[0143] Compound LN175-57: Under nitrogen protection, bromoethanol (0.12 g / 1.0 mmol) and potassium carbonate (0.20 g / 1.5 mmol) were added to a DMF (10 mL) solution containing N-26 (0.97 g / 1.0 mmol). The internal temperature was maintained at 0-5 °C during the addition process. After the addition was complete, the mixture was stirred at room temperature for 5 hours. Bromoethanol (0.12 g / 1.0 mmol) was added again, and the reaction was continued for 12 hours. The mixture was filtered, and ice water (30 mL) was added. The mixture was extracted with dichloromethane (30 mL × 3), and the organic phase was washed with water (30 mL). The residue was concentrated and purified by pre-HPLC to obtain 0.23 g of compound LN175-57 (23% yield). MS: 1012.3 [M+1]. 1 H NMR(DMSO-d6)δ9.16-9.15(m,1H),7.74-7.72(m,2H),7.35-7.34(m,1H),7. 03-7.01(m,1H),6.44-6.40(m,2H),4.84-4.79(m,5H),4.03-4.01(m,1H),3 .56-3.45(m,6H),3.21-3.18(m,2H),2.94-2.89(m,3H),2.57-2.41(m,3H), 2.34-2.30(m,1H),1.63-1.54(m,6H),1.35-1.32(m,1H),1.05-1.03(m,1H).

[0144] Example 27: Preparation and characterization of compounds LN175-58 and LN175-59

[0145] 0.27 g of compound LN175-58 (25% yield) was prepared from N-26 using the method of Example 19. MS: 1092.0 [M+1]. 1 H NMR(DMSO-d6)δ9.12-9.11(m,1H),7.72-7.71(m,2H),7.36-7.35(m,1H),7.02-7.01(m,1H),6.44-6.42(m,2H),4.83-4.80(m,5H),4.04-4.02 (m,1H),3.45-3.41(m,4H),2.93-2.90(m,3H),2.58-2.44(m,3H),2.31 -2.29(m,1H),1.64-1.55(m,6H),1.37-1.30(m,3H),1.03-1.00(m,1H).

[0146] 19.3 mg of compound LN175-59 (17% yield) was prepared from LN175-58 as the starting material using the method of Example 21. MS: 1158.2 [M+23]. 1 H NMR(DMSO-d6)δ9.14-9.13(m,1H),7.73-7.71(m,2H),7.38-7.36(m,1H),7.04-7.02(m,1H),6.48-6.46(m,2H),4.85-4.81(m,5H),4.03-4.00 (m,1H),3.47-3.41(m,4H),2.96-2.91(m,3H),2.59-2.45(m,3H),2.33 -2.29(m,1H),1.63-1.53(m,6H),1.33-1.31(m,3H),1.04-1.02(m,1H).

[0147] Example 28: Preparation and characterization of compound LN175-60

[0148] 0.28 g of compound LN175-60 (27% yield) was prepared from N-26 using the method of Example 22. MS: 1070.1 [M+1]. 1 H NMR(DMSO-d6)δ9.18-9.17(m,1H),7.78-7.77(m,2H),7.43-7.42(m,1H),7 .08-7.05(m,1H),6.54-6.50(m,2H),4.89-4.82(m,5H),4.09-4.05(m,1H) ,3.78(s,3H),3.51-3.42(m,4H),2.99-2.92(m,3H),2.62-2.48(m,3H),2. 34-2.30(m,1H),1.68-1.52(m,6H),1.32-1.30(m,3H),1.03-1.00(m,1H).

[0149] Example 29: Preparation and Characterization of Compound N-31

[0150] 11.3 g of compound N-31 was synthesized according to the method of Example 10. MS: 971.1 [M+1].

[0151] Example 30: Preparation and characterization of compound LN175-61

[0152] Compound LN175-61: Under nitrogen protection, water (2 mL) and sodium hydroxide (0.08 g / 2.0 mmol) were added to a 20 mL solution of N-31 (0.97 g / 1.0 mmol). After the addition was complete, the mixture was stirred for 10 hours, filtered, concentrated, and the resulting solid was recrystallized from acetone / water (2:1) to give 0.18 g of compound LN175-61 (18% yield). MS: 1015.3 [M+23]. 1 H NMR(DMSO-d6)δ9.14-9.13(m,1H),7.73-7.72(m,2H),7.33-7.32(m,1H),7.03-7.02(m,1H),6.44-6.42(m,2H),4.83-4.79(m,4H),4 .03-4.01(m,1H),3.50-3.43(m,4H),2.57-2.46(m,3H),2.33-2.29(m,1H),1.66-1.54(m,6H),1.33-1.31(m,1H),1.02-0.99(m,1H).

[0153] Example 31: Preparation and characterization of compounds LN175-63 and LN175-64

[0154] 0.15 g of compound LN175-63 (14% yield) was prepared from N-31 using the method of Example 19. MS: 1081.2 [M+1]. 1 H NMR(DMSO-d6)δ9.17-9.16(m,1H),7.75-7.74(m,2H),7.35-7.33(m,1H),7.06-7.05(m,1H),6.46-6.43(m,2H),5.87(s,2H),4.85-4.81(m, 4H),4.04-4.00(m,1H),3.57-3.44(m,4H),2.59-2.47(m,3H),2.36-2 .31(m,1H),1.65-1.55(m,6H),1.32-1.31(m,1H),1.05-1.02(m,1H).

[0155] 9.0 mg of compound LN175-64 (8% yield) was prepared from LN175-63 as the starting material using the method of Example 21. MS: 1147.3 [M+23]. 1H NMR(DMSO-d6)δ9.16-9.15(m,1H),7.74-7.72(m,2H),7.36-7.34(m,1H),7.07-7.05(m,1H),6.47-6.45(m,2H),5.83(s,2H),4.86-4.80(m, 4H),4.03-4.01(m,1H),3.56-3.45(m,4H),2.56-2.48(m,3H),2.33-2 .30(m,1H),1.66-1.57(m,6H),1.34-1.32(m,1H),1.03-1.01(m,1H).

[0156] Example 32: Preparation and characterization of compound LN175-65

[0157] 0.37 g of compound LN175-65 (35% yield) was prepared using the method of Example 22 starting material N-31. MS: 1059.1 [M+1]. 1 H NMR(DMSO-d6)δ9.19-9.18(m,1H),7.78-7.76(m,2H),7.36-7.34(m,1H) ,7.08-7.05(m,1H),6.45-6.42(m,2H),5.95(s,2H),4.86-4.82(m,4H),4 .07-4.05(m,1H),3.78(s,3H),3.59-3.48(m,4H),2.56-2.46(m,3H),2.3 8-2.36(m,1H),1.64-1.54(m,6H),1.34-1.32(m,1H),1.04-1.02(m,1H).

[0158] Example 33: Preparation and characterization of compound LN175-67

[0159] 0.32 g of compound LN175-67 (30% yield) was prepared using the method of Example 22 with N-31 as the starting material. MS: 1073.2 [M+1]. 1H NMR(DMSO-d6)δ9.18-9.17(m,1H),7.77-7.76(m,2H),7.33-7.32(m,1H),7.04-7.03(m,1H),6.43-6.41(m,2H),5.93-5.91(m,1H),4.85-4.83 (m,4H),4.06-4.05(m,1H),3.60-3.47(m,4H),2.55-2.47(m,3H),2.36 -2.35(m,1H),1.65-1.52(m,9H),1.32-1.30(m,1H),1.03-1.02(m,1H).

[0160] Example 34: Preparation and Characterization of Compound N-33

[0161] 13.5 g of compound N-33 was synthesized according to the method of Example 10. MS: 971.4 [M+1].

[0162] Example 35: Preparation and characterization of compound LN175-70

[0163] 10.0 mg of compound LN175-70 (10% yield) was prepared from N-33 using the method of Example 30. MS: 1015.3 [M+23]. 1 H NMR(DMSO-d6)δ9.12-9.11(m,1H),7.71-7.70(m,2H),7.31-7.30(m,1H),7.02-7.01(m,1H),6.43-6.41(m,2H),4.84-4.79(m,4H),4 .04-4.02(m,1H),3.52-3.44(m,4H),2.93-2.91(m,3H),2.32-2.30(m,1H),1.67-1.55(m,6H),1.33-1.30(m,1H),1.02-1.00(m,1H).

[0164] Example 36: Preparation and characterization of compounds LN175-72 and LN175-73

[0165] 0.14 g of compound LN175-72 (13% yield) was prepared from N-33 using the method of Example 19. MS: 1081.0 [M+1]. 1H NMR(DMSO-d6)δ9.14-9.12(m,1H),7.71-7.69(m,2H),7.30-7.28(m,1H),7.01-7.00(m,1H),6.42-6.40(m,2H),5.80(s,2H),4.83-4.80(m, 4H),4.03-4.01(m,1H),3.54-3.45(m,4H),2.94-2.92(m,3H),2.34-2 .31(m,1H),1.62-1.53(m,6H),1.31-1.29(m,1H),1.01-1.00(m,1H).

[0166] 12.4 mg of compound LN175-73 (11% yield) was prepared from LN175-72 using the method of Example 21. MS: 1147.1 [M+23]. 1 H NMR(DMSO-d6)δ9.13-9.12(m,1H),7.70-7.69(m,2H),7.31-7.28(m,1H),7.03-7.01(m,1H),6.44-6.41(m,2H),5.82(s,2H),4.84-4.81(m, 4H),4.00-3.98(m,1H),3.56-3.48(m,4H),2.96-2.93(m,3H),2.32-2 .30(m,1H),1.61-1.54(m,6H),1.32-1.30(m,1H),1.02-1.00(m,1H).

[0167] Example 37: Preparation and characterization of compound LN175-74

[0168] 0.26 g of compound LN175-74 (25% yield) was prepared from N-33 using the method of Example 22. MS: 1059.3 [M+1]. 1 H NMR(DMSO-d6)δ9.15-9.14(m,1H),7.74-7.73(m,2H),7.32-7.31(m,1H) ,7.04-7.03(m,1H),6.47-6.45(m,2H),5.96(s,2H),4.86-4.83(m,4H),4 .02-3.99(m,1H),3.73(s,3H),3.57-3.46(m,4H),2.95-2.93(m,3H),2.3 4-2.32(m,1H),1.63-1.55(m,6H),1.34-1.32(m,1H),1.03-1.02(m,1H).

[0169] Example 38: Preparation and characterization of compounds LN175-76 and LN175-77

[0170] 0.13 g of compound LN175-76 (12% yield) was prepared from N-31 as the starting material according to the method of Example 11. MS: 1081.2 [M+1]. 1 H NMR(DMSO-d6)δ9.16-9.15(m,1H),7.73-7.72(m,2H),7.33-7.31(m,1H),7.03-7.02(m,1H),6.43-6.42(m,2H),5.73(s,2H),4.83-4.80(m, 4H),4.03-4.01(m,1H),3.56-3.46(m,4H),2.55-2.44(m,3H),2.33-2 .31(m,1H),1.62-1.53(m,6H),1.31-1.30(m,1H),1.04-1.03(m,1H).

[0171] 11.2 mg of compound LN175-77 (10% yield) was prepared from LN175-76 as the starting material according to the method of Example 13. MS: 1147.3 [M+23]. 1 H NMR(DMSO-d6)δ9.13-9.12(m,1H),7.72-7.70(m,2H),7.33-7.31(m,1H),7.03-7.02(m,1H),6.44-6.43(m,2H),5.76(s,2H),4.83-4.80(m, 4H),4.02-4.00(m,1H),3.54-3.44(m,4H),2.57-2.49(m,3H),2.32-2 .31(m,1H),1.62-1.55(m,6H),1.30-1.29(m,1H),1.01-1.00(m,1H).

[0172] Example 39: Preparation and Characterization of Compound LN175-78

[0173] 0.33 g of compound LN175-78 (31% yield) was prepared from N-31 as the starting material according to the method of Example 15. MS: 1059.1 [M+1]. 1H NMR(DMSO-d6)δ9.15-9.14(m,1H),7.71-7.70(m,2H),7.30-7.29(m,1H) ,7.01-7.00(m,1H),6.42-6.40(m,2H),5.79(s,2H),4.82-4.79(m,4H),4 .03-4.01(m,1H),3.74(s,3H),3.55-3.46(m,4H),2.58-2.55(m,3H),2.3 1-2.28(m,1H),1.61-1.54(m,6H),1.31-1.29(m,1H),1.02-1.01(m,1H).

[0174] Example 40: Preparation and characterization of compounds LN175-79 and LN175-80

[0175] 0.22 g of compound LN175-76 (20% yield) was prepared from N-33 as the starting material according to the method of Example 11. MS: 1081.1 [M+1]. 1 H NMR(DMSO-d6)δ9.12-9.10(m,1H),7.70-7.69(m,2H),7.31-7.30(m,1H),7.03-7.02(m,1H),6.43-6.41(m,2H),5.79(s,2H),4.85-4.81(m, 4H),4.02-4.01(m,1H),3.55-3.46(m,4H),2.92-2.89(m,3H),2.31-2 .30(m,1H),1.60-1.52(m,6H),1.30-1.29(m,1H),1.02-1.00(m,1H).

[0176] 14.6 mg of compound LN175-80 (13% yield) was prepared from LN175-79 as the starting material according to the method of Example 13. MS: 1147.0 [M+23]. 1 H NMR(DMSO-d6)δ9.14-9.12(m,1H),7.73-7.72(m,2H),7.30-7.29(m,1H),7.02-7.01(m,1H),6.44-6.42(m,2H),5.75(s,2H),4.83-4.81(m, 4H),4.03-4.02(m,1H),3.54-3.45(m,4H),2.93-2.90(m,3H),2.32-2 .30(m,1H),1.61-1.53(m,6H),1.32-1.30(m,1H),1.01-0.99(m,1H).

[0177] Example 41: Preparation and characterization of compound LN175-81

[0178] 0.16 g of compound LN175-81 (15% yield) was prepared from N-33 as the starting material according to the method of Example 15. MS: 1059.3 [M+1]. 1 H NMR(DMSO-d6)δ9.15-9.14(m,1H),7.75-7.74(m,2H),7.30-7.29(m,1H),7.03-7.02(m,1H),6.43-6.41(m,2H),5.79(s,2H),4.84-4.82(m, 4H),4.05-4.03(m,1H),3.52-3.46(m,4H),2.94-2.92(m,3H),2.31-2 .29(m,1H),1.62-1.54(m,6H),1.33-1.31(m,1H),1.02-1.01(m,1H).

[0179] The compounds in the following examples were prepared using a similar synthetic method:

[0180] Example 42: In vitro anti-HIV activity test of the compound

[0181] Test compounds: control compound Lenacapavir and test compounds LN175-01 to LN175-81.

[0182] Experimental materials: MT-4 cells, HIV-1 wild-type strain (III) B ), HIV-2 strain (ROD).

[0183] Preparation of test compounds: Accurately weigh an appropriate amount of the test compound, dissolve it thoroughly in DMSO, and then dilute it with double-distilled water to prepare 6 concentration gradients (50 nM, 100 nM, 200 nM, 400 nM, 600 nM, 800 nM).

[0184] Culture and sample loading: Add MT-4 cells (1×10⁶ cells) sequentially to the 96-well plate. 5 50 μL of culture medium, 20 μL of HIV-infected MT-4 cell suspension or 20 μL of HIV-uninfected MT-4 cell suspension (for CC) 50Different concentrations of the analyte or control compound were used for determination, with final concentrations controlled at 50 nM, 100 nM, 200 nM, 400 nM, 600 nM, and 800 nM. The mixtures were then incubated at 37°C in a 5% CO2 incubator for 5 consecutive days. After incubation, 20 mL of 5 mg / mL MTT solution was added to each well, and the mixture was incubated for another 2 hours. After centrifugation at 100 rpm for 10 min, the supernatant was removed, and 150 μL of DMSO was added to each well. The mixture was shaken at low speed for 30 min, and the absorbance was measured at 540 nm using an ELISA reader. Three replicates were used for each concentration.

[0185] Data calculation:

[0186] Based on the absorbance measured at different concentrations of the test compound, the cell proliferation rate P1% (Equation 1) and cell death rate P2% (Equation 2) were calculated. Then, linear regression equations were established based on the logarithm of the test compound concentration and the cell proliferation rate and cell death rate at different concentrations. The data were processed using GraphPad Prism 8, and the curves of cell proliferation rate and concentration and the curves of cell death rate and concentration were fitted to obtain the results.

[0187] Formula 1: Cell proliferation rate P1% = (Absorbance of test compound group - Absorbance of infected group) / Absorbance of infected group × 100%

[0188] Equation 2: Cell mortality rate P2% = (Absorbance of blank group - Absorbance of test compound group) / Absorbance of blank group × 100%

[0189] EC was calculated based on the relationship curve between cell proliferation rate and concentration. 50 The CC value was calculated based on the cell death rate and concentration relationship curve. 50 value.

[0190] EC 50 CC 50 The data is shown in Table 1:

[0191] Table 1. EC5 cell activity of compounds 50 CC 50 value

[0192] Compared to Lenacapavir (EC 50 =389 nM), the compound provided by this invention exhibits better HIV-1 antiviral activity (EC). 50 =70-380nM), of which LN175-04, LN175-26, LN175-34, LN175-46, LN175-61, LN175-65 and LN175-75 have EC50 The values ​​were even below 100 nM, demonstrating strong antiviral activity; simultaneously, in terms of antiviral activity against HIV-2 strains, LN175-04, LN175-26, LN175-34, LN175-46, LN175-61, LN175-65, and LN175-75 showed high EC50 values. 50 Values ​​between 50 and 100 nM also exhibited strong antiviral activity, showing a significant improvement compared to Lenacapavir. Based on the above experimental data, compounds LN175-04, LN175-26, LN175-34, LN175-46, LN175-61, LN175-65, and LN175-75 of this invention demonstrate a clear advantage in in vitro anti-HIV activity.

[0193] Example 43: In vitro assay of the compound's inhibitory activity against HIV-1 reverse transcriptase (HIV-1RT)

[0194] Experimental materials:

[0195] HIV-1 RT kit; test compounds LN175-04, LN175-26, LN175-34, LN175-46, LN175-61, LN175-65 and LN175-75; control compound Lenacapavir.

[0196] Experimental principle:

[0197] In reverse transcription, a template of Poly(A) was selected, with oligo(dT)15 as the primer and biotin and digoxigenin-labeled dNTPs as the substrates to synthesize a DMA molecule labeled with both. This DNA molecule can bind to a microplate coated with streptavidin. Then, peroxidase conjugated with digoxigenin antibody and ABTS (peroxidase substrate) were added sequentially, resulting in a color change. The activity value (IC50) was... 50 The absorbance value (A) can be calculated from the absorbance value detected by the microplate reader.

[0198] Experimental groups: Sample group, negative group (no inhibitor, with HIV-1RT) and blank control group (no inhibitor, no HIV-1RT).

[0199] Experimental method: The experiment was conducted according to the instructions of the HIV-1RT kit.

[0200] Prepare a 0.1 ng / μL enzyme solution by dissolving 250 ng of HIV-1 reverse transcriptase in lysis buffer and triple-distilled water. Dissolve the test compound or control compound Lenacapavir in DMSO and dilute with lysis buffer to prepare six concentration gradients (1.6 nM, 8 nM, 40 nM, 200 nM, 1000 nM, 5000 nM), with three replicates for each concentration.

[0201] Take 20 μL each of the pre-prepared enzyme solution, the test compound solution or the control compound solution, and the mixed reaction solution containing DNA template, primers and dNTPs. Perform PCR amplification according to the standard operating procedure in the kit instructions, and then add them to the PCR reaction tube and incubate at 37°C for 2 h.

[0202] Add the above-mentioned incubated sample solution to a 96-well plate coated with streptavidin, and incubate again at 37°C for 2 hours;

[0203] Rinse the wells with buffer (5 × 250 μL), then add peroxidase-linked digoxigenin antibody solution (200 μL / well) and incubate again for 2 h;

[0204] Wash the plate again with buffer (5 × 250 μL), then add ABTS substrate solution (200 μL / well) and incubate for 2 h;

[0205] The absorbance (A) of the sample in each well plate was measured at a wavelength of 405 nm using an ELISA reader.

[0206] The inhibition rate (%) was calculated according to Equation 3:

[0207] Formula 3: Inhibition rate % = (OD value of negative control group (without inhibitor, with HIV-1RT) - OD value of inhibitor group (with inhibitor and HIV-1RT)) / (OD value of negative control group (without inhibitor, with HIV-1RT) - OD value of blank group (without inhibitor and HIV-1RT)) × 100%

[0208] Then, a linear regression equation is established with the logarithm of the concentration to calculate the IC. 50 The values ​​and results are shown in Table 2:

[0209] Table 2. Inhibitory activity of compounds against HIV-1 WT RT

[0210] Compared with Lenacapavir, the compounds LN175-04, LN175-26, LN175-34, LN175-46, LN175-61, LN175-65 and LN175-75 of the present invention all showed significant inhibitory activity against HIV reverse transcription, achieving unexpected results.

[0211] Example 44: Pharmacokinetics of the compound in animals

[0212] Laboratory animals:

[0213] Thirty male Beagles weighing 8-10 kg were randomly divided into 10 groups of 3 dogs each. Groups 1-5 were administered the drug orally via gavage, while groups 6-10 were administered the drug intravenously. The dogs had free access to food and water and were kept at a room temperature of 20-26℃, humidity of 40-70%, and a light intensity of 12h:12h before the experiment was conducted.

[0214] Test sample preparation:

[0215] Preparation of oral administration sample: Weigh a certain amount of sample, add 5% ethanol, 20% PG, 45% PEG300 and 30% deionized water (adjust pH to about 2 with 0.01N HCl) to the final volume, and mix thoroughly by sonication to obtain an administration solution with a concentration of 0.8 mg / mL.

[0216] Preparation of intravenous injection sample: Weigh a certain amount of sample, add 5% DMSO, 30% PG, 30% PEG400% and 35% physiological saline to the final volume, and mix thoroughly by sonication to obtain an administration solution with a concentration of 0.5 mg / mL.

[0217] Administration:

[0218] The experimental animals were fasted from 8 PM the day before administration. The following morning at 8 AM, each group of animals was administered the drug via gavage, subcutaneous injection, or intravenous injection, respectively. Feeding began 4 hours after administration, and water was allowed throughout the experiment. The administration protocol is shown in Table 3.

[0219] Table 3 Dosing regimens for pharmacokinetic testing

[0220] Blood collection via gavage administration: Within 0.5 hours before gavage administration (0h), and at 0.5h, 1.5h, 3.0h, 4.5h, 6.0h, 8.0h, 10.0h, 24.0h, and 48.0h after administration, approximately 1.0mL of blood was collected from the forelimb vein and placed in an EDTA-K2 anticoagulant blood collection tube. The plasma was separated by centrifugation (1000rpm, 10min, 2-8℃) and stored at -70℃.

[0221] Blood collection for intravenous administration: Within 0.5 hours before a single intravenous injection (0h), and at 0.15h, 0.5h, 1.0h, 3.0h, 5.0h, 8.0h, 10.0h, 24.0h, and 48.0h after administration, approximately 1mL of blood is collected from the forelimb vein and placed in an EDTA-K2 anticoagulant blood collection tube. The plasma is separated by centrifugation (1000rpm, 10min, 2-8℃) and stored at -70℃.

[0222] Take 50 μL of plasma sample, add 200 μL of methanol, mix by sonication, centrifuge (1000 rpm, 10 min), take 5 μL of supernatant for injection, and determine the blood drug concentration by LC-MS / MS.

[0223] Calculate the bioavailability (%) of the oral administration according to Formula 4:

[0224] Formula 4: Bioavailability (%) of gavage administration = (AUC) inf-ig / dosage dose) / (AUC) inf-iv / dosage dose) × 100%.

[0225] The average pharmacokinetic parameters for injection administration are shown in Table 4, and the average pharmacokinetic parameters for oral administration are shown in Table 5.

[0226] Table 4. Average pharmacokinetic parameters for injection administration (n=3)

[0227] Table 5. Average pharmacokinetic parameters after gavage administration (n=3)

[0228] The main pharmacokinetic parameter AUC of the test compound was compared with that of the control compound Lenacapavir. Inf (exposure), C max (Peak blood concentration), t 1 / 2 Both half-life and bioavailability were significantly improved, exhibiting superior pharmacokinetic characteristics. Specifically, the bioavailability of LN175-26 and LN175-46 was more than 15% higher than that of Lenacapavir, with AUC... Inf Increased by nearly double, C max Increase by 2 to 3 times, t 1 / 2 The extension was significant, achieving unexpected technical results.

[0229] Example 45: Acute toxicity test of the compound in vivo

[0230] One hundred healthy KM mice (Kunming mice) were acclimatized for one week in an environment with a temperature of 25±5℃ and a humidity of 55±5%, with free access to water and food. Forty mice weighing 20±2g were then randomly divided into five groups of eight each: control group, LN175-26 group, LN175-46 group, LN175-61 group, and LN175-75 group. A quantitative amount of the compound was weighed and prepared into a suspension using 5% DMSO / 70% water / 25% PEG400. The compound was administered by gavage at a dose of 1.0g / kg. The control group received no medication but only the same volume of the 5% DMSO / 70% water / 25% PEG400 solution, with free access to water and food. The administration continued for 15 days, with body weight measured every three days. The mathematical mean body weight of each group was used as the overall body weight data. The general physiological condition of the mice was also observed. The body weight data are shown in Table 6.

[0231] Table 6: Changes in mouse body weight at a dose of 1.0 g / kg

[0232] Experimental data show that, when administered orally at a dose of 1.0 g / kg, there was no significant difference in weight gain between the experimental and control groups. Furthermore, no mice died during the experiment, indicating that the compounds LN175-26, LN175-46, LN175-61, and LN175-75 provided in this patent have low acute toxicity and good safety in mice, consistent with in vitro test data. Significant beneficial technical effects have been achieved.

[0233] During the experiment, the general physiological condition of the mice was observed. The control group mice and the mice in the LN175-26, LN175-46, LN175-61 and LN175-75 groups did not show loss of appetite or lethargy throughout the experiment. Throughout the experiment, all groups of mice had dull fur, were agile, and were in good mental condition, indicating that the compound was expected to have small toxic side effects in vivo.

[0234] Through the above detailed description of this patent, those skilled in the art can thoroughly understand the features of this invention, and the improvements made to this invention also fall within the scope of the appended claims.

Claims

1. Compounds of Formula I: Including its solvates, isotopic derivatives, or salts thereof; in: Y1 and Y2 are each independently selected from CH or N; R1 and R2 are each independently selected from hydrogen, halogen, hydroxyl, cyano, and C. 1-6 Alkyl, C 1-6 Alkoxy; Ring A is selected from five- to six-membered heterocycles having one to three nitrogen atoms and carbon atoms, and is optionally substituted with R3 or R4; R3 and R4 are each independently selected from hydrogen, amino, hydroxyl, and C. 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 Carbocyclic group, deuterated C 1-6 Alkyl, -NHR9, -OR9, The aforementioned alkyl, alkoxy, and carbocyclic groups may optionally be substituted with one or more hydrogen, halogen, trifluoromethyl, cyano, methanesulfonyl, ethanesulfonyl, isopropanesulfonyl, cyclopropanesulfonyl, or p-toluenesulfonyl groups; T is selected from hydrogen, sodium, potassium, Hydroxyethyl; R w1 R w2 R w3 Each is independently selected from hydrogen and C. 1-6 alkyl; R x R y Each is independently selected from hydrogen, metal ions, and C. 1-6 Alkyl, C 3-6 carbon cyclo group, C 2-10 Heterocyclic group, C 3-10 Heterocyclic aryl, or R x R y Linked into a ring; the above-mentioned heterocyclic groups and heterocyclic aryl groups may optionally be substituted by one or more hydrogens, halogens, hydroxyl groups, amino groups, nitro groups, methyl groups, ethyl groups, isopropyl groups, carboxyl groups, methoxy groups, ethoxy groups, isopropoxy groups, methylamino groups, ethylamino groups, isopropylamino groups, mercapto groups, mercaptomethyl groups, mercaptoethyl groups, acetyl groups, trifluoromethyl groups, methanesulfonyl groups, and ethanesulfonyl groups; R z Selected from C 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 carbon cyclo group, C 2-10 Heterocyclic group, C 3-10 Heterocyclic aryl, deuterated C 1-6 Alkyl; the above-mentioned alkyl, alkoxy, carbocyclic, heterocyclic, and heterocyclic aryl groups may optionally be substituted by one or more hydrogen, halogen, hydroxyl, amino, nitro, methyl, ethyl, isopropyl, carboxyl, methoxy, ethoxy, isopropoxy, methylamino, ethylamino, isopropylamino, mercapto, mercaptomethyl, mercaptoethyl, acetyl, trifluoromethyl, methanesulfonyl, and ethanesulfonyl groups; R5 is selected from hydrogen, halogen, hydroxyl, cyano, C 1-6 Alkyl, C 1-6 Alkoxy; R6 is selected from C 1-6 Alkyl, C 1-6 Alkoxy, C 2-8 alkenyl, C 2-8 Alkyne group, deuterated C 2-8 Alkynyl; the above-mentioned alkyl, alkoxy, alkenyl, and alkynyl groups may optionally be substituted with one or more hydrogen, cyano, trifluoromethyl, methanesulfonyl, ethanesulfonyl, isopropanesulfonyl, cyclopropanesulfonyl, p-toluenesulfonyl, deuterated methanesulfonyl, deuterated ethanesulfonyl, deuterated isopropanesulfonyl, deuterated cyclopropanesulfonyl, deuterated formyl, or deuterated acetyl. R7 is selected from C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 alkylamine group, C 3-6 Carbocyclic group; the above-mentioned alkyl, alkoxy, alkylamine, and carbocyclic groups may optionally be converted by one or more R groups. 10 replace; R8 is selected from C 1-6 Alkyl, C 3-6 Carbocyclic group; the above-mentioned alkyl or carbocyclic group may optionally be converted by one or more R groups. 11 replace; R9 is selected from formyl, acetyl, propionyl, cyclopropionyl, benzoyl, methanesulfonyl, ethanesulfonyl, isopropanesulfonyl, cyclopropanesulfonyl, p-toluenesulfonyl, hydroxyethyl, deuterated methanesulfonyl, deuterated ethanesulfonyl, deuterated isopropanesulfonyl, deuterated cyclopropanesulfonyl, deuterated formyl, and deuterated acetyl. R 10 Selected from C 2-10 Heterocyclic group, C 6-15 Aromatic ring group, C 3-10 Heterocyclic aryl; the above-mentioned heterocyclic group, aryl group, heterocyclic aryl group may optionally be substituted by one or more hydrogen, halogen, hydroxyl, amino, nitro, methyl, ethyl, isopropyl, carboxyl, methoxy, ethoxy, isopropoxy, methylamino, ethylamino, isopropylamino, mercapto, mercaptomethyl, mercaptoethyl, acetyl, trifluoromethyl, methanesulfonyl, ethanesulfonyl; R 11 Selected from C 2-10 Heterocyclic group, C 3-10 Heterocyclic aryl; the above-mentioned heterocyclic groups and heterocyclic aryl groups may optionally be substituted by one or more hydrogens, halogens, hydroxyl groups, amino groups, nitro groups, methyl groups, ethyl groups, isopropyl groups, carboxyl groups, methoxy groups, ethoxy groups, isopropoxy groups, methylamino groups, ethylamino groups, isopropylamino groups, mercapto groups, mercaptomethyl groups, mercaptoethyl groups, acetyl groups, trifluoromethyl groups, methanesulfonyl groups, and ethanesulfonyl groups; G is selected from hydrogen, Hydroxyethyl; When T is selected from hydrogen; G is selected from Hydroxyethyl; When T is selected from sodium or potassium; G is selected from hydrogen, and R9 is selected from deuterated methanesulfonyl, deuterated ethanesulfonyl, deuterated isopropanesulfonyl, deuterated cyclopropanesulfonyl, deuterated formyl, or deuterated acetyl. T is selected from hour; G is selected from hydrogen; Furthermore, T and G are not both selected from hydrogen; R a R b Each is independently selected from hydrogen and C. 1-6 Alkyl, or R a R b Connected to form a ring; R c R d Each is independently selected from hydrogen and C. 1-6 Alkyl, or R c R d Connected to form a ring; R 12 R 13 Each is independently selected from hydrogen, metal ions, and C. 1-6 Alkyl, C 3-6 carbon cyclo group, C 2-10 Heterocyclic group, C 3-10 Heterocyclic aryl, or R 12 R 13 Linked into a ring; the above-mentioned heterocyclic groups and heterocyclic aryl groups may optionally be substituted by one or more hydrogens, halogens, hydroxyl groups, amino groups, nitro groups, methyl groups, ethyl groups, isopropyl groups, carboxyl groups, methoxy groups, ethoxy groups, isopropoxy groups, methylamino groups, ethylamino groups, isopropylamino groups, mercapto groups, mercaptomethyl groups, mercaptoethyl groups, acetyl groups, trifluoromethyl groups, methanesulfonyl groups, and ethanesulfonyl groups; R 14 Selected from C 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 carbon cyclo group, C 2-10 Heterocyclic group, C 3-10 Heterocyclic aryl, deuterated C 1-6 Alkyl; the above-mentioned alkyl, alkoxy, carbocyclic, heterocyclic, and heterocyclic aryl groups may optionally be substituted with one or more hydrogens, halogens, hydroxyl groups, amino groups, nitro groups, methyl groups, ethyl groups, isopropyl groups, carboxyl groups, methoxy groups, ethoxy groups, isopropoxy groups, methylamino groups, ethylamino groups, isopropylamino groups, mercapto groups, mercaptomethyl groups, mercaptoethyl groups, acetyl groups, trifluoromethyl groups, methanesulfonyl groups, and ethanesulfonyl groups.

2. The compound of claim 1, having the structure of formula II: The substituents in Formula II are defined as defined in Formula I.

3. The compound of claim 1, having the structure of formula III: The substituents in Formula III are defined as defined in Formula I.

4. The compound of claim 1, having the following structure:

5. A pharmaceutical composition comprising the compound of claims 1-4 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

6. Use of the compounds of claims 1-4 and / or the pharmaceutical composition of claim 5 in the preparation of an anti-HIV agent.

7. A method for treating an individual infected with HIV, comprising administering to the mammal to be treated a therapeutically effective amount of the compound of claims 1-4 and / or the pharmaceutical composition of claim 5.

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