Baloxavir derivative and use thereof

Abstract

Provided are a baloxavir derivative and use thereof. The baloxavir derivative has the structural formula shown in formula 1, wherein the sEH inhibitor is a residue of an esterified soluble epoxide hydrolase inhibitor. The use is use of the baloxavir derivative or a pharmaceutical composition thereof in the preparation of an anti-inflammatory and anti-viral medicament. By linking a series of sEH inhibitors to BAX to synthesize a twin drug, the present invention can inhibit the NF-κB pathway and target virus cap-dependent endonucleases, thereby exerting the anti-inflammatory and anti-viral effects.

Description

A Baloxavir Derivative and Its Uses

[0001] Priority claim: This invention claims priority to Chinese Patent Application No. 2024116795411, filed on November 22, 2024, the entire contents of which are incorporated herein by reference. Technical Field

[0002] This invention belongs to the field of pharmaceutical science and relates to a baloxavir derivative and its uses. Background Technology

[0003] Seasonal influenza is an acute respiratory infectious disease caused by the influenza virus, characterized by high infectivity and rapid spread, and is a major global public health problem. Most hospitalizations and deaths are attributed to complications such as pulmonary fibrosis, bacterial pneumonia, acute respiratory distress syndrome (ARDS), and congestive heart failure. In the early stages of influenza virus infection, influenza can induce upregulation of TLR3 protein levels, activating the NF-κB signaling pathway and mitochondrial-associated oxidative stress, thereby triggering an inflammatory response in the host's immune system to eliminate the virus. However, the production of inflammatory mediators does not decrease with the reduction of viral load, leading to an excessive accumulation of inflammatory cytokines in the later stages of infection, which may cause further damage to the host. Currently approved anti-influenza drugs do not address the excessive inflammatory response in the host, highlighting the urgent need for strategies to mitigate this response during viral infection. High-dose corticosteroids have been shown to alleviate lung damage in patients with severe influenza pneumonia; however, they may also reduce viral clearance and prolong recovery time. The concurrent use of antiviral and anti-inflammatory drugs raises concerns about potential metabolic side effects, including liver and kidney damage. Furthermore, patients often exhibit poor adherence when multiple medications are required. Therefore, the applicant proposed a novel approach that combines antiviral and anti-inflammatory compounds into a single entity to achieve both therapeutic effects in treating influenza virus infection.

[0004] Baloxavir marboxil (MBAX) is currently the first single-dose oral antiviral drug approved in the EU for the treatment of uncomplicated influenza and post-exposure prophylaxis in individuals aged 1 year and older. The active form, baloxavir, inhibits influenza virus transcription by blocking the endonuclease activity of the PA protein. The keto-enol structure of baloxavir (BAX) contributes to its hydrophilicity, resulting in a very low rate of intestinal membrane transport. To improve the bioavailability of baloxavir, the hydroxyl group in the keto-enol structure is replaced, forming baloxavir ester.

[0005] Arachidonic acid (AA) metabolized by cytochrome P450 enzymes, EETs, are closely related to inflammation. Studies have found that inhibition of NF-κB activity is key to the anti-inflammatory effect of EpFAs, including EETs. EETs inhibit NF-κB activity through three pathways: first, by inhibiting IκB kinase to suppress the activation of NF-κB by tumor necrosis factor (TNF)-α; second, by activating PPARγ to inhibit the transcription of NF-κB and AP-1; and third, by directly inhibiting the activity of the downstream inflammatory cytokine PGE2 in the NF-κB signaling pathway, which is an effect not found in NSAIDs and steroidal anti-inflammatory drugs. EETs produced by amino acids (AA) through the CYP450 metabolic pathway have many physiological functions, such as anti-inflammation, analgesia, anti-ischemia, anti-angiogenesis, anti-proliferation, cardiovascular protection, and regulation of metabolic diseases. EETs, together with pro-inflammatory metabolites such as prostaglandins and leukotrienes from the COX and LOX metabolic pathways of AA, form the body's pro-inflammatory / anti-inflammatory balance, playing a very important role in maintaining the body's inflammatory / anti-inflammatory balance. sEH inhibitors can stabilize endogenous EpFAs (EETs), therefore, sEH is a potential therapeutic target for pain and inflammatory diseases.

[0006] A search revealed no studies on the use of sEH inhibitors linked to BAX to synthesize twin drugs that simultaneously inhibit the NF-κB pathway and target viral cap-dependent endonucleases to exert dual anti-inflammatory and antiviral effects. Summary of the Invention

[0007] This invention synthesizes twin drugs by linking a series of sEH inhibitors with BAX to simultaneously inhibit the NF-κB pathway and target viral cap-dependent endonucleases, thereby exerting a dual anti-inflammatory and antiviral effect. Novel compounds with anti-inflammatory and antiviral activities were identified in vitro and in vivo.

[0008] To achieve the above objectives, the present invention adopts the following technical solution.

[0009] In a first aspect, the present invention provides a baloxavir derivative, its stereoisomer, its crystalline hydrate, its deuterated derivative, its solvate, its prodrug, or a pharmaceutically acceptable salt thereof.

[0010] [Amended according to Rule 26, 25.02.2026] The structural formula of the baloxavir derivative is shown in Formula I:

[0011] Among them, sEH inhibitor is the esterified residue of a soluble epoxide hydrolase inhibitor.

[0012] Preferably, the soluble epoxide hydrolase inhibitor contains a substituted benzene ring or a substituted adamantane.

[0013] Preferably, the soluble epoxide hydrolase inhibitor is t-AUCB, 12-{[(tricyclic[3.3.1.1 3，7 [dec-1-ylamino]carbonyl]amino}dodecanoic acid (AUDA), (S)-1-(4-(3-(1r,3R,5S,7S)-3,5-dimethyladamantane-1-yl)ureo)-3-fluorobenzyl)piperidine-3-carboxylic acid, 7-{[(tricyclic[3.3.1.1 3，7 [dec-1-ylamino]carbonyl]amino}heptanoic acid, 4-(((1r,4r)-4-(2-(3-fluoro-4-(trifluoromethoxy)phenyl)acetamido)cyclohexyl)oxy)benzoic acid, t-TUCB, 12-({[(1,3-dimethyltricyclo[3.3.1.1] 3，7 ]dec-7-yl)amino]carbonyl}amino)dodecanoic acid, 12-{[({4-[(trifluoromethyl)oxy]phenyl}amino)carbonyl]amino}dodecanoic acid, 12-{[({3-fluoro-4-[(trifluoromethyl)oxy]phenyl}amino)carbonyl]amino}dodecanoic acid, 11-{[(tricyclo[3.3.1.1 3，7 [dec-1-ylamino]carbonyl]amino}undecanoic acid, 11-({[(3,5-dimethyltricyclo[3.3.1.1]) 3，7 [dec-1-yl)amino]carbonyl}amino)undecanoic acid, 8-{[(tricyclic[3.3.1.1] 3，7 [dec-3-ylamino]carbonyl]amino}octanoic acid, with 12-[({[1-(tricyclic[3.3.1.1] 3，7 [dec-1-yl]ethyl]amino]carbonyl]amino]dodecanoic acid, 12-[({[1-(tricyclic[3.3.1.1] 3，7 At least one of 12-([(bicyclo[2.2.1]hept-2-en-6-ylmethyl)amino]carbonyl)amino)dodecanoic acid, decanoic acid, and 12-({[(bicyclo[2.2.1]hept-2-en-6-ylmethyl)amino]carbonyl)amino)dodecanoic acid).

[0014] In some embodiments, the compound represented by Formula I is selected from the following compounds:

[0015] In a second aspect, the present invention provides a pharmaceutical composition comprising the above-mentioned baloxavir derivative, its stereoisomer, its crystalline hydrate, its deuterated derivative, its solvate, its prodrug, or a pharmaceutically acceptable salt thereof.

[0016] Preferably, the pharmaceutical composition further includes traditional Chinese medicine ingredients and / or Western medicine ingredients.

[0017] Preferably, the pharmaceutical composition further includes a pharmaceutically acceptable carrier.

[0018] Preferably, the pharmaceutical composition is a tablet, pill, cream, emulsion, ointment, suspension, lyophilized agent, capsule, sustained-release agent, granule, powder, injectable agent, or spray.

[0019] Thirdly, the present invention provides the use of the above-mentioned baloxavir derivatives, their stereoisomers, their crystalline hydrates, their deuterated derivatives, their solvates, their prodrugs, their pharmaceutically acceptable salts, or pharmaceutical compositions in the preparation of anti-inflammatory and antiviral drugs.

[0020] Preferably, the virus is an influenza virus.

[0021] More preferably, the influenza virus is influenza A virus and / or influenza B virus.

[0022] More preferably, the influenza virus is at least one of H1N1, H5N1, H7N9, H3N2 and influenza B virus.

[0023] More preferably, the influenza virus is at least one of H1N1, H3N2, or type B influenza virus.

[0024] Fourthly, the present invention provides the use of the above-mentioned baloxavir derivatives, their stereoisomers, their crystalline hydrates, their deuterated derivatives, their solvates, their prodrugs, or their pharmaceutically acceptable salts or pharmaceutical compositions thereof in the preparation of NF-κB signaling pathway inhibitors.

[0025] The anti-inflammatory and antiviral drugs or NF-κB signaling pathway inhibitors described in this invention are suitable for humans or animals.

[0026] Preferably, the animals include bovines, equines, sheep, pigs, canines, felines, rabbits, birds, rodents, or primates.

[0027] Beneficial effects of the present invention

[0028] (1) This invention synthesizes twin drugs by linking a series of sEH inhibitors with BAX to simultaneously inhibit the NF-κB pathway and target viral cap-dependent endonucleases to exert dual anti-inflammatory and antiviral effects, and identifies new compounds with anti-inflammatory and antiviral activities in vitro and in vivo.

[0029] (2) Compared with BAX and MBAX, the baloxavir derivative of the present invention has better anti-influenza virus activity.

[0030] (3) Clinically widely used antiviral drugs, such as oseltamivir and baloxavir, are recommended to be administered within 48 hours of infection to effectively protect influenza patients. However, the baloxavir derivative of this invention still has a good protective effect when administered within 48-96 hours after influenza virus infection. Attached Figure Description

[0031] Figure 1 shows the inhibitory effects of MBAX, ATV-114, and AUDA on H1N1 virus protein expression in cell lines with high and low AADAC enzyme expression.

[0032] Figure 2 shows the anti-inflammatory activity of ATV-114 by inhibiting NF-κB signaling. In Figure 2, A represents the inhibitory effect of different concentrations of MBAX, ATV-114, and AUDA on LPS-induced expression of inflammatory proteins in RAW264.7 cells, and B represents the inhibition of LPS-induced NF-κB transcriptional activation in RAW264.7 cells transfected with NF-κB luciferase reporter plasmid by different concentrations of MBAX, ATV-114, and AUDA.

[0033] Figure 3 shows how ATV-114 prevents the nuclear output of RNP by inhibiting the nuclear translocation of NF-κB.

[0034] Figure 4 shows the anti-influenza virus effect. In Figure 4, A represents the body weight change of mice 14 days after oral administration 12 hours after infection with A / PR / 8 / 34; B represents the survival of mice 14 days after oral administration 12 hours after infection with A / PR / 8 / 34; C represents the body weight change of mice 14 days after oral administration at different time points (48, 72, and 96 hours) after infection with A / PR / 8 / 34; D represents the survival of mice 14 days after oral administration at different time points (48, 72, and 96 hours) after infection with A / PR / 8 / 34; E represents the body weight change of mice 14 days after oral administration 12 hours after infection with A / PR / 8 / 34 with the NA-H274Y strain; and F represents the body weight change of mice 14 days after oral administration 12 hours after infection with A / PR / 8 / 34 with the NA-H274Y strain. Detailed Implementation

[0035] Terminology Definition

[0036] Unless otherwise stated, the following terms and phrases used in this invention have the following meanings:

[0037] In this invention, "room temperature" refers to ambient temperature, ranging from 10°C to 40°C. In some embodiments, "room temperature" refers to a temperature ranging from 20°C to 30°C; in other embodiments, "room temperature" refers to a temperature ranging from 25°C to 30°C; and in still other embodiments, "room temperature" refers to 10°C, 15°C, 20°C, 25°C, 30°C, 35°C, 40°C, etc.

[0038] In this invention, "esterified residues" refers to the residual fragments after the carboxyl group of a soluble epoxide hydrolase inhibitor forms an ester bond with the hydroxyl group of baloxavir.

[0039] Baloxavir derivatives

[0040] "Stereoisomers" are compounds that have the same chemical structure but whose atoms or groups are arranged differently in space. Stereoisomers include enantiomers, diastereomers, conformational isomers (rotational isomers), geometric isomers (cis / trans) isomers, and hindered isomers.

[0041] The compounds represented by Formula I in this invention may have a chiral center, such as a chiral carbon. The compounds therefore comprise racemic mixtures of all stereoisomers, including enantiomers, diastereomers, and transisomers. Additionally, the compounds of this invention comprise optical isomers enriched or resolved at any or all of the asymmetric chiral atoms. In other words, the chiral center, similar to that described, is provided in the form of chiral isomers or racemic mixtures. Mixtures of racemic and diastereomers, as well as isolated or synthesized individual optical isomers substantially free of their enantiomers or diastereomer conjugates, are all within the scope of this invention. Racemic mixtures are isolated into their individual, substantially optically pure isomers using known techniques, such as isolating salts of diastereomers formed with optically active auxiliaries (e.g., acids or bases) and then converting them back to the optically active substance. In most cases, the desired optical isomer is synthesized from a suitable stereoisomer of the desired starting material through a stereospecific reaction.

[0042] "Hydrate" refers to an assemblage formed when the solvent molecules are water.

[0043] "Deuteration" refers to the substitution of one or more hydrogen atoms in a compound or group by deuterium. Deuteration can be monosubstituted, disubstituted, polysubstituted, or total substituted.

[0044] "Solvate" refers to an association formed by one or more solvent molecules with the compound of the present invention. Solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, dimethyl sulfoxide, ethyl acetate, acetic acid, and aminoethanol.

[0045] The term "prodrug" is used in its broadest sense and includes those derivatives that are converted into the compounds of this invention in vivo. These derivatives are readily apparent to those skilled in the art. In prodrug design, the most common form is the structural modification of drugs containing carboxyl, hydroxyl, or amino groups to produce prodrugs of the types of esters, carboxylic acid esters, amino acid esters, amides, and phosphate esters. In addition, azo prodrugs, nitric oxide prodrugs, and prodrugs with novel structures such as open-ring and closed-ring prodrugs can also be produced.

[0046] Preferred prodrugs are those that, relative to the parent substance, improve the bioavailability of the compounds of the invention when used on a patient (e.g., by making orally administered compounds more readily absorbed into the bloodstream) or enhance the delivery of the parent compound to biological compartments (brain or lymphatic system).

[0047] The compounds of this invention may be in crystalline form as a beneficial compound or as a solvator, both of which are included within the scope of this invention. Methods of solvation are well known in the art. Suitable solvators are pharmaceutical solvators.

[0048] "Pharmaceutical acceptable" means a molecular entity or composition that is physiologically tolerable when administered to a human and generally does not produce allergic or similar undesirable reactions, such as gastrointestinal upset, dizziness, etc. Preferably, as used in this invention, "pharmaceutical acceptable" means a substance approved by a federal regulatory agency or national government or listed in the Chinese Pharmacopoeia, the United States Pharmacopoeia, or other generally recognized pharmacopoeias for use in animals, and more particularly in humans.

[0049] "Pharmaceutically acceptable salts" include salts derived from suitable bases, such as alkali metals or alkaline earth metals (e.g., Na₂O₃). + Li + K + Ca +2 and Mg +2 ) and ammonium. Physiologically acceptable salts of nitrogen atoms or amino groups include: (a) acid addition salts formed with inorganic acids, such as hydrochloric acid, hydrobromic acid, sulfuric acid, aminosulfonic acid, phosphoric acid, nitric acid, etc.; (b) salts formed with organic acids, such as acetic acid, oxalic acid, tartaric acid, succinic acid, maleic acid, fumaric acid, gluconic acid, citric acid, malic acid, ascorbic acid, benzoic acid, ethanesulfonic acid, lactobionic acid, tannic acid, palmitic acid, alginic acid, polyglutamic acid, naphthalene. Sulfonic acids, methanesulfonic acids, p-toluenesulfonic acids, benzenesulfonic acids, naphthalenedisulfonic acids, polygalacturonic acid, malonic acid, sulfosalicylic acid, glycolic acid, 2-hydroxy-3-naphthylcarboxylate, dihydroxynaphthylcarboxylate, salicylic acid, stearic acid, phthalic acid, mandelic acid, lactic acid, ethanesulfonic acid, lysine, arginine, glutamic acid, glycine, serine, threonine, alanine, isoleucine, leucine, etc.; and (c) salts formed with elemental anions, such as chlorine, bromine, and iodine. Physiologically acceptable salts of hydroxyl compounds include salts formed with anions of the compounds such as Na+. + and NR4 + A suitable combination of cations.

[0050] For therapeutic purposes, "acceptable" as used herein means physiologically acceptable, i.e., salts derived from physiologically acceptable acids or bases. However, salts that are not physiologically acceptable acids or bases may also be used, for example, to prepare or purify physiologically acceptable compounds. All salts, whether or not derived from physiologically acceptable acids or bases, are within the scope of this invention.

[0051] Compounds of Formula I or their pharmaceutically acceptable salts may exist as different polymorphs or pseudopolymorphs. Crystal polymorphism, as used in this invention, refers to the ability of a crystalline compound to exist in different crystal structures. Crystal polymorphism can originate from differences in crystal packing (packing polymorphism) or differences in packing between different conformational isomers of the same molecule (conformational polymorphism). Pseudopolymorphism, as used in this invention, refers to the ability of a compound's hydrates or solvates to exist in different crystal structures. Pseudopolymorphs of this invention can exist due to differences in crystal packing (packing pseudopolymorphism) or due to differences in packing between different conformational isomers of the same molecule (conformational pseudopolymorphism). This invention encompasses all polymorphs and pseudopolymorphs of compounds of Formula I and their pharmaceutically acceptable salts.

[0052] Compounds of Formula I or their pharmaceutically acceptable salts can also exist as amorphous solids. The amorphous solids used in this invention are solids in which the positions of atoms do not exhibit long-range order. This definition also applies when the crystal size is 2 nanometers or less. Additives, including solvents, can be used to establish the amorphous forms of this invention. This invention encompasses all amorphous forms of compounds of Formula I and their pharmaceutically acceptable salts.

[0053] "Post-esterification residues" are the remaining carboxylic acid fragments after the carboxyl group of the soluble epoxide hydrolase inhibitor forms an ester bond with the hydroxyl group of baloxavir.

[0054] "Substitution" refers to the independent replacement of one or more hydrogen atoms in a benzene ring or adamantane by non-hydrogen substituents.

[0055] Screening of compositions with anti-inflammatory and antiviral activities

[0056] The baloxavir derivatives described in this invention are suitable for the treatment or prevention of influenza virus infection in animals or humans. However, in the process of screening compounds that can resist inflammation and influenza viruses, cell-based and animal (rodent) assays are the main screening tools.

[0057] The compositions of the present invention are screened against baloxavir derivatives having anti-inflammatory and antiviral activities using any conventional techniques for evaluating anti-inflammatory and antiviral activities. In the context of the present invention, typically, compositions having anti-inflammatory and antiviral activities are first screened, followed by screening for in vivo activity of compositions exhibiting anti-inflammatory and antiviral activities. The composition having less than about 5 × 10⁻⁶ ppm... -6 M is preferably less than about 1×10 -7 Compositions for the in vitro Ki (inhibition constant) of M are preferably used in vivo. Useful in vitro screening methods have been described in detail in the literature and will not be repeated here. However, the examples describe suitable in vitro assays.

[0058] Pharmaceutical Composition

[0059] The pharmaceutical compositions described in this invention comprise the aforementioned baloxavir derivatives and pharmaceutically acceptable excipients. In some embodiments, the baloxavir derivatives described in this invention are provided in the pharmaceutical composition in an effective amount (e.g., a therapeutically effective amount).

[0060] The pharmaceutical compositions of this invention are formulated with conventional carriers and excipients, which will be selected according to conventional practice. Although the active ingredients can be administered alone, it is preferred to formulate them into pharmaceutical preparations. The preparations of this invention, whether for animal or human use, comprise at least one active ingredient as defined above and one or more acceptable carriers therefor, and optionally include other therapeutic ingredients, particularly those disclosed herein. The carrier must be "acceptable," meaning compatible with other components in the preparation and physiologically harmless to its recipient.

[0061] The pharmaceutical compositions described herein can be prepared by any method known in pharmaceutical science. Generally, these methods of preparation involve associating the baloxavir derivative (i.e., the “active ingredient”) with a carrier or excipient and / or one or more other auxiliary ingredients, and then, if desired and / or expected, shaping and / or packaging the product into desired single-dose or multi-dose units.

[0062] The pharmaceutical compositions of the present invention can be prepared according to known methods, such as those described in the General Rules for Preparation of Pharmaceuticals in the Chinese Pharmacopoeia, the 16th edition of the Japanese Pharmacopoeia, the United States Pharmacopoeia, and the 9th edition of the European Pharmacopoeia. Depending on the dosage form, the pharmaceutical compositions of the present invention can be appropriately administered to patients.

[0063] Pharmaceutical compositions may be prepared, packaged, and / or sold in bulk, as a single unit dose, and / or as multiple single unit doses. A “unit dose” is a discrete amount of a pharmaceutical composition containing a predetermined amount of the active ingredient. The amount of the active ingredient is typically equal to the dose of the active ingredient administered to a subject and / or a convenient fraction of that dose (e.g., one-half or one-third of the dose).

[0064] The active ingredients and pharmaceutically acceptable excipients in the pharmaceutical compositions described herein will vary depending on the identity, body type, and / or condition of the treated subject and further on the route of administration of the composition. The pharmaceutical composition may contain between 0.1% and 100% (w / w) of the active ingredient.

[0065] Pharmaceutically acceptable excipients used in the preparation of the provided pharmaceutical compositions include inert diluents, dispersants and / or granulators, surfactants and / or emulsifiers, disintegrants, binders, preservatives, buffers, lubricants and / or oils. The pharmaceutical compositions may also contain excipients (e.g., cocoa butter and suppository waxes), colorants, coating agents, sweeteners, flavoring agents, and aroma agents.

[0066] The present invention further provides a veterinary composition comprising at least one active ingredient as defined above and a veterinary carrier thereon.

[0067] The veterinary carrier is a substance intended for use in veterinary compositions and can be a solid, liquid, or gaseous substance. Furthermore, it is inert or acceptable in the veterinary field and compatible with the active ingredient. These veterinary compositions can be administered orally, parenterally, or via any other desired route.

[0068] As used herein, the term "treatment" means, unless otherwise stated, reversing or alleviating the condition or disease to which the term applies, or one or more symptoms of such condition or disease, inhibiting the progression of said condition or disease or one or more symptoms thereof, or preventing said condition or disease or one or more symptoms thereof. As used herein, the term "treatment" refers to a therapeutic act, as defined above.

[0069] The “effective amount” of the compounds described in this invention refers to an amount sufficient to elicit the desired biological response. The effective amount of the compounds described in this invention may vary depending on factors such as the desired biological endpoint, the pharmacokinetics of the compound, the condition being treated, the administration method, and the age and health status of the subject. In some embodiments, the effective amount is a therapeutically effective amount. Alternatively, in a single method or use, the invention may be used for prophylactic treatment when indicated and effective. In some embodiments, the effective amount is the amount of the compound described herein in a single dose. In some embodiments, the effective amount is a combination of the compound described herein in multiple doses.

[0070] The term "therapeuticly effective amount" as used herein refers to an amount sufficient to provide therapeutic benefit in the treatment of a condition or sufficient to delay or minimize one or more symptoms associated with that condition. For baloxavir derivatives, the therapeutically effective amount means the amount by which the therapeutic agent, alone or in combination with other therapies, provides therapeutic benefit in the treatment of a condition. The term "therapeuticly effective amount" may encompass amounts that improve overall therapy, reduce or prevent symptoms, signs, or causes of a condition, and / or enhance the therapeutic efficacy of another therapeutic agent. In some embodiments, the therapeutically effective amount is an amount sufficient to treat any of the described diseases or conditions.

[0071] Application route

[0072] One or more compounds of the present invention (referred to herein as active ingredients) may be administered via any route suitable for the condition being treated. Suitable routes include oral, rectal, nasal, pulmonary, local (including oral and sublingual), and parenteral (including subcutaneous, intramuscular, intravenous, intradermal, intrathecal, and epidural). It should be understood that preferred routes may vary depending on, for example, the recipient's condition. The advantage of the compounds of the present invention is that they are orally bioavailable and can be administered orally.

[0073] It should also be noted that the specific dosage and method of administration of the baloxavir derivative for different patients are determined by many factors, including the patient's age, weight, sex, natural health condition, nutritional status, drug activity, timing of administration, metabolic rate, severity of illness, and the subjective judgment of the treating physician. The effective dose of the active ingredient depends at least on the nature of the condition to be treated, its toxicity (regardless of whether the compound is used prophylactically or against an active viral infection), the method of delivery, and the pharmaceutical formulation, and will be determined through clinicians using routine dose escalation studies. Expected doses are approximately 0.0001 to approximately 100 mg / kg body weight per day; typically, approximately 0.01 to approximately 10 mg / kg body weight per day; more typically, approximately 0.01 to approximately 5 mg / kg body weight per day; and most typically, approximately 0.05 to approximately 0.5 mg / kg body weight per day. For example, for an adult weighing approximately 70 kg, the candidate daily dose would be in the range of 1 mg to 1000 mg, preferably 5 mg to 500 mg, and could be administered in single or multiple doses.

[0074] To enable those skilled in the art to better understand the technical solutions of the present invention, some non-limiting embodiments are further disclosed below to provide a more detailed description of the present invention.

[0075] All reagents used in this invention can be purchased commercially or prepared by the methods described in this invention.

[0076] Example 1: 4-[(4-{[(three rings[3.3.1.1)] 3,7 [dec-1-ylamino]carbonyl]amino}cyclohexyl]oxy]benzoic acid-{[(12aR)-12-[(11S)-7,8-difluoro-6,11-dihydrodibenzo[1,2-e:1',2'-b]thiacycloheptam-11-yl]-6,8-dioxane-1,3,4,6,12,12a-hexahydro[1,4]oxazacyclohexam[3,4-c]pyrido[2,1-f][1,2,4]triazacyclohexam-7-yl]oxy}methyl ester (ATV-103)

[0077] At 0℃, 4-[(4-{[(tricyclic[3.3.1.1 3，7[decyl-1-ylamino]carbonyl]amino]cyclohexyl]oxy]benzoic acid t-AUCB (1.0 g, 2.42 mmol), sodium bicarbonate (1.01 g, 12.1 mmol), and tetrabutylammonium bisulfate (82.1 mg, 0.242 mmol) were dissolved in 20 mL of a 1:1 mixture of dichloromethane and water. Chloromethyl chlorosulfonate (0.97 mL, 3.36 mmol) was added dropwise. After addition, the mixture was stirred at room temperature for 8 h. The organic layer was separated, and the aqueous layer was extracted with 50 mL of dichloromethane. The extract and organic layer were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by rapid silica gel chromatography (0-50% ethyl acetate / petroleum ether) to give intermediate 1 (0.88 g, 79.1% yield).

[0078] Baloxavir (0.5 g, 1.03 mmol), intermediate 1 (0.57 g, 1.24 mmol), and potassium carbonate (0.35 g, 2.57 mmol) were dissolved in 15 mL of acetonitrile and reacted at 80 °C for 8 h. After cooling the reaction solution to room temperature, the crude product was concentrated and purified by rapid silica gel chromatography (0-80% ethyl acetate / petroleum ether) to give compound ATV-103 (0.56 g, yield 59.3%). 1 H NMR (600MHz, DMSO-d6) δ7.92-7.86 (m, 2H), 7.43-7.35 (m, 2H), 7.22 (d, J= 7.8Hz, 1H), 7.18-7.14(m, 1H), 7.13-7.08(m, 1H), 7.08-7.05(m, 2H), 7.03 -7.00 (m, 1H), 6.88-6.84 (m, 1H), 6.00 (d, J = 6.5Hz, 1H), 5.94 (d, J = 6.5Hz , 1H), 5.73 (d, J=7.8Hz, 1H), 5.62 (d, J=7.6Hz, 1H), 5.40 (d, J=4.0Hz, 1H), 4.48-4.42(m, 1H), 4.42-4.38(m, 1H), 4.27-4.20(m, 1H), 4.08-4.04(m, 1 H), 3.97 (dd, J=10.9, 3.1Hz, 1H), 3.49 (dd, J=11.3, 3.2Hz, 1H), 3.43-3.35 (m, 2H), 297-2.91 (m, 1H), 2.88-2.81 (m, 1H), 2.05-1.96 (m, 6H), 1.85 (d, J =2.9Hz, 8H), 1.59 (q, J = 3.1Hz, 6H), 1.49-1.41 (m, 2H), 1.28-1.19 (m, 3H). 13C NMR (151MHz, DMSO-d6) δ173.6, 170.8, 165.3, 162.2, 156.9, 156.9, 154.5, 151.9, 150.4, 150.3, 150. 2, 148.0, 147.9, 146.4, 146.3, 141.2, 136.4, 134.2, 132.7, 132.1, 130.2, 129.4, 128.6, 127.8, 127.0 , 125.5, 125.1, 125.0, 121.7, 117.0, 116.9, 115.7, 114.0, 88.7, 75.0, 73.7, 69.8, 68.8, 65.9, 60.2, 49.8, 47.2, 46.0, 42.5, 42.5, 40.5, 36.6, 31.4, 30.7, 30.1, 29.4, 26.8, 23.6, 22.6, 21.2, 14.6, 14.4.

[0079] Example 2: 12-{[(three rings[3.3.1.1 3，7 [dec-1-ylamino]carbonyl]amino}dodecanoic acid-{[(12aR)-12-[(11S)-7,8-difluoro-6,11-dihydrodibenzo[1,2-e:1′,2′-b]thiacycloheptan-11-yl]-6,8-dioxane-1,3,4,6,12,12a-hexahydro[1,4]oxazacyclohexane[3,4-c]pyrido[2,1-f][1,2,4]triazacyclohexane-7-yl]oxy}methyl ester (ATV-114)

[0080] According to the method described in Example 1, 12-{[(three rings[3.3.1.1]} 3，7 Using dec-1-ylamino)carbonyl]amino}dodecanoic acid (AUDA) as a starting material, compound ATV-114 was synthesized as a white solid (0.3 g), with an overall yield of 45.6% in both steps. 1H NMR (600MHz, DMSO-d6) δ7.43-7.3 (m, 2H), 7.22 (d, J=7.7Hz, 1H), 7.15 (t, J=7.6Hz, 1H), 7.10 (d, J=7.9Hz, 1H), 7.02 (d, J=7.7H z, 1H), 6.85 (t, J=7.4Hz, 1H), 5.73-5.69 (m, 3H), 5.58 (t, J=5.6Hz, 1H), 5.42 (d, J=3.1Hz, 1H), 4.47-4.37 (m, 2H), 4.06 (d, J=1 4.3Hz, 1H), 3.99 (dd, J=11.0, 3.0Hz, 1H), 3.67 (dd, J=11.7, 3.3Hz, 1H), 3.47 (t, J=10.4Hz, 1H), 3.32 (s, 1H), 3.31-3.26 (m, 1H ), 2.98-2.86 (m, 3H), 2.31 (t, J=7.4Hz, 2H), 2.00-1.95 (m, 3H), 1.84 (d, J=2.9Hz, 6H), 1.62-1.50 (m, 8H), 1.34-1.16 (m, 16H). 13 C NMR (151) MHz, DMSO-d6) δ173.7, 172.9, 157.5, 154.6, 150.6, 150.3, 141.2, 136.4, 1 34.2, 132.7, 130.2, 129.3, 128.6, 127.8, 127.0, 125.4, 125.2, 125.1, 117 .0, 116.9, 113.9, 88.3, ​​73.7, 69.8, 68.9, 66.2, 49.8, 46.0, 42.5, 40.5, 39 .2, 36.6, 34.0, 30.5, 29.5, 29.4, 29.4, 29.4, 29.3, 28.9, 26.9, 24.5, 23.6.

[0081] Example 3: (S)-1-(4-(3-(1r,3R,SS,7S)-3,5-dimethyladamantane-1-yl)ureo)-3-fluorobenzyl)piperidine-3-carboxylic acid (intermediate 5)

[0082] 4-(bromomethyl)-2-fluoro-1-nitrobenzene (3.0 g, 12.9 mmol) and dry acetonitrile (35 mL) were added to a single-necked flask. After the 4-(bromomethyl)-2-fluoro-1-nitrobenzene dissolved, potassium carbonate (2.13 g, 15.4 mmol), potassium iodide (0.15 g, 1.29 mmol), and (S)-piperidine-3-carboxylic acid ethyl ester (2.03 g, 12.9 mmol) were added, and the mixture was refluxed. The reaction was monitored by TLC after 6 h to ensure completion. Acetonitrile was removed by vacuum concentration, water (20 mL) was added, and the mixture was extracted with ethyl acetate (30 mL × 2). The organic layers were combined, washed with water (20 mL), and then washed once with saturated brine (25 mL). The organic phase was concentrated under reduced pressure to obtain 5.2 g of yellow oil. The mixture was packed into a 4x silica gel column, mixed with 1.2x silica gel, and eluent (EA:PE = 1:40). Column chromatography yielded a yellow oil, which was (S)-1-(3-fluoro-4-nitrobenzyl)piperidine-3-carboxylic acid ethyl ester (intermediate 2), with a yield of 4.4 g. The yield was 82%.

[0083] Ethyl (S)-1-(3-fluoro-4-nitrobenzyl)piperidine-3-carboxylate (3.60 g, 11.6 mmol), 5% Pd-C (0.4 g), and anhydrous ethanol (50 mL) were added to a single-necked flask. The mixture was purged twice with argon and twice with hydrogen. The temperature was raised to 60 °C and the mixture was stirred for 12 h. The reaction was monitored by TLC until it was complete. After the reaction solution was cooled to room temperature (25 °C), it was filtered. The filtrate was concentrated under reduced pressure to obtain a yellow oily substance, which was ethyl (S)-1-(4-amino-3-fluorobenzyl)piperidine-3-carboxylate (intermediate 3). The yield was 2.86 g, and the yield was 86%.

[0084] Add solid phosgene (1.06 g, 3.8 mmol) and dry DCM (30 mL) to a three-necked flask. Cool the flask to below -78 °C with cold hydrazine. Add dropwise a solution of (S)-1-(4-amino-3-fluorobenzyl)piperidine-3-carboxylic acid ethyl ester (2.13 g, 7.6 mmol) and triethylamine (1.06 g, 3.980 mmol) in dry dichloromethane (60 mL). After the addition is complete, move the flask to room temperature and stir for 0.5 h. Then stop the reaction. Concentrate the resulting reaction solution to dryness under reduced pressure. Dissolve the residue in dry DCM (10 mL) to obtain an isocyanate solution for later use.

[0085] Add memantine (1.48 g, 7.6 mmol), triethylamine (1.54 g, 15.2 mmol), and dry dichloromethane (25 mL) to a single-necked flask. Add the above isocyanate solution dropwise and react at room temperature for 0.5 h. TLC shows that the reaction is complete. Pour the reaction solution into water (40 mL), extract with EA (40 mL × 3), then wash successively with 1 mol / L HCl (40 mL × 2), water (40 mL × 2), saturated brine (40 mL), and dry with anhydrous sodium sulfate. Filter and concentrate the filtrate under reduced pressure to obtain 3.4 g of a pale yellow oil, which is (S)-1-(4-(3-(1r,3R,5S,7S)-3,5-dimethyladamantane-1-yl)ureo)-3-fluorobenzyl)piperidine-3-carboxylic acid ethyl ester (intermediate 4), which is unpurified and directly added to the next step.

[0086] Add (S)-1-(4-(3-(1r,3R,5S,7S)-3,5-dimethyladamantane-1-yl)ureo)-3-fluorobenzyl)piperidine-3-carboxylic acid ethyl ester (intermediate 4) (3.4 g, 7.0 mmol), dissolved in tetrahydrofuran (20 mL), sodium hydroxide (0.36 g, 9.00 mmol), and water (10 mL) to a single-necked flask, and reflux. TLC showed complete reaction after 12 h; the reaction solution was concentrated under reduced pressure to remove tetrahydrofuran, the residue was added with water (300 mL), placed in a cold trap, and the pH was adjusted to 3 with 6N hydrochloric acid (10 mL), precipitating a pale yellow solid. The solid was filtered, the filter cake was washed with water (20 mL), and dried to obtain 2.96 g of crude product (dried in an oven at 60 °C for 24 h). The crude product was purified by slurrying with petroleum ether and diethyl ether to obtain a pale yellow solid, which is (S)-1-(4-(3-(1r,3R,5S,7S)-3,5-dimethyladamantane-1-yl)ureo)-3-fluorobenzyl)piperidine-3-carboxylic acid (intermediate 5), yielding 2.8 g, with a yield of 85.5%.

[0087] Example 4: (3S)-1-{[4-({[(1,3-dimethyltricyclo[3.3.1.1] 3，7 [dec-5-yl)amino]carbonyl}amino)-3-fluorophenyl]carbonyl}hexahydropyridine-3-carboxylic acid-{[(12aR)-12-[(11S)-7,8-difluoro-6,11-dihydrodibenzo[1,2-e:1′,2′-b]thiacycloheptan-11-yl]-6,8-dioxane-1,3,4,6,12,12a-hexahydro[1,4]oxazacyclohexane[3,4-c]pyrido[2,1-f][1,2,4]triazacyclohexane-7-yl]oxy}methyl ester (ATV-116)

[0088] According to the method described in Example 1, using (S)-1-(4-(3-(1r,3R,5S,7S)-3,5-dimethyladamantane-1-yl)ureo)-3-fluorobenzyl)piperidine-3-carboxylic acid (intermediate 5) as the starting material, 0.41 g of compound ATV-116 was synthesized as a white solid, with an overall yield of 47.1% in both steps. 1 H NMR (600MHz, DMSO-d6) δ8.29 (d, J=3.2Hz, 1H), 8.20 (t, J=8.3Hz, 1H), 7.43 -7.35(m, 2H), 7.27-7.17(m, 2H), 7.17-7.07(m, 3H), 7.00(s, 1H), 6.83(s, 1H), 6.57 (s, 1H), 5.70 (d, J=7.9Hz, 2H), 5.42 (d, J=14.4Hz, 1H), 4.47-4.2 8 (m, 2H), 4.06 (d, J = 14.3Hz, 1H), 4.00-3.94 (m, 1H), 3.65 (d, J = 11.2Hz, 1H ), 3.47 (d, J=10.4Hz, 1H), 3.31 (s, 2H), 3.11-3.04 (m, 1H), 2.91 (s, 1H), 2. 62-2.56 (m, 1H), 2.11-2.07 (m,, 1H), 2.05-2.00 (m, 1H), 1.76 (d, J=3.2Hz, 2H), 1.72-1.63(m, 2H), 1.58(s, 4H), 1.48(d, J=13.3Hz, 1H), 1.36-1.29(m , 3H), 1.29-1.22 (m, 4H), 1.18 (t, J=7.1Hz, 1H), 1.12 (s, 2H), 0.83 (s, 6H). 13 C NMR (151MHz, DMSO-d6) δ173.6, 172.5, 168.5, 154.5, 153.8, 150.4, 150.3, 150.2, 150.1, 146.4, 1 41.1, 136.3, 134.2, 132.7, 130.3, 130.2, 129.3, 128.7, 128.7, 128.5, 127.9, 127.0, 125.4, 125.1 , 123.9, 119.1, 117.0, 116.9, 114.4, 114.3, 113.9, 88.4, 73.8, 69.8, 68.9, 66.3, 60.2, 52.1, 50.7, 48.0, 45.9, 42.8, 41.2, 40.5, 40.5, 35-3, 32.4, 31.6, 30.5, 30.3, 30.0, 27.0, 26.8, 23.6, 22.6.

[0089] Example 5: 7-{[(three rings[3.3.1.1 3，7 [dec-1-ylamino]carbonyl]amino}heptanoic acid

[0090] Add solid phosgene (1.06 g, 3.8 mmol) and dry DCM (30 mL) to a single-necked flask. Cool the flask to below -78 °C and add a solution of adamantane (1.15 g, 7.6 mmol) and triethylamine (1.06 g, 3.980 mmol) in dry dichloromethane (60 mL). After the addition is complete, move the flask to room temperature and stir for 0.5 h. Then stop the reaction. Concentrate the resulting reaction solution to dryness under reduced pressure. Add dry DCM (10 mL) to the residue to dissolve it and obtain an isocyanate solution for later use.

[0091] 1.21 g (7.6 mmol) of methyl 7-aminoheptanate, 1.54 g (15.2 mmol) of triethylamine, and 25 mL of dry dichloromethane were added to a three-necked flask. The above isocyanate solution was added dropwise, and the reaction was allowed to proceed at room temperature for 0.5 h. TLC showed that the reaction was complete. The reaction solution was poured into water (40 mL), extracted with EA (40 mL × 3), and then washed successively with 1 mol / L HCl (40 mL × 2), water (40 mL × 2), saturated brine (40 mL), and dried over anhydrous sodium sulfate. The solution was filtered, and the filtrate was concentrated under reduced pressure to obtain 2.5 g of a pale yellow oil, which was methyl 7-(3-(3s,5s,7s)-adamantane-1-yl)ureoyl)heptanate (intermediate 6). It was not purified and was directly added to the next step.

[0092] Methyl 7-(3-(3s,5s,7s)-adamantane-1-yl)ureoyl)heptanoate (intermediate 6) (3.4 g, 10.1 mmol) was added to a single-necked flask, dissolved in tetrahydrofuran (20 mL), sodium hydroxide (0.48 g, 12.1 mmol), and water (10 mL). The mixture was refluxed. After 12 h, TLC showed that the reaction was complete. The reaction solution was concentrated under reduced pressure to remove tetrahydrofuran. The residue was added to water (300 mL), placed in a cold trap, and the pH was adjusted to 3 with 6N hydrochloric acid (10 mL). A pale yellow solid precipitated. The solid was filtered, and the filter cake was washed with water (20 mL) to obtain 7-{[(tricyclic[3.3.1.1] 3，7 [dec-1-ylamino]carbonyl]amino}heptanoic acid (intermediate 7) yielded 2.8 g, with a yield of 86.0%.

[0093] Example 6: 7-{[(three rings[3.3.1.1 3，7[dec-1-ylamino]carbonyl]amino}heptanoic acid-{[(12aR)-12-[(11S)-7,8-difluoro-6,11-dihydrodibenzo[1,2-e:1′,2′-b]thiacycloheptan-11-yl]-6,8-dioxane-1,3,4,6,12,12a-hexahydro[1,4]oxazacyclohexane[3,4-c]pyrido[2,1-f][1,2,4]triazacyclohexane-7-yl]oxy}methyl ester (ATV-117)

[0094] According to the method described in Example 1, 7-{[(three rings[3.3.1.1 3，7 Using dec-1-ylamino)carbonyl]amino}heptanoic acid (intermediate 7) as a starting material, compound ATV-117 was synthesized in a total of 0.56 g of white solid, with an overall yield of 52.6% in both steps. 1 H NMR (600MHz, DMSO-d6) δ7.44-7.37 (m, 2H), 7.22 (d, J=7.8Hz, 1H), 7.18-7.13 (m, 1H), 7.09 (dd, J=8.0, 1.3Hz, 1H), 7.02 (dd, J=8.0, 1.5Hz, 1H), 6.87-6.83 (m, 1H), 5.73-5.68 (m, 4H), 5.60 (t, J=5.7Hz, 1H), 5.43 (d, J=13.8Hz, 2H), 4.47-4.39 (m, 2H), 4.06 (d, J=14.3Hz, 1H), 3.99 (dd, J=10.9, 3.1Hz, 1H), 3.68 (dd, J=11.6, 3.3Hz, 1H), 3.48 (t, J=10.4Hz, 1H), 3.31-3.26 (m, S 1H), 2.98-2.88(m, 3H), 2.34-2.29(m, 2H), 1.99-1.94(m, 3H), 1.83(d, J=2.9Hz, 6H), 1.62-1.51(m, 8H), 1.34-1.22(m, 6H). 13C NMR(151 MHz, DMSO-d6) δ173.7, 172.9, 157.5, 154.6, 152.0, 151.9, 150.6, 150.3, 150.2, 14 8.0, 147.9, 146.4, 141.2, 136.4, 134.2, 132.7, 132.7, 130.2, 129.3, 128.5, 127.8 , 127.0, 127.0, 125.4, 125.2, 125.1, 117.1, 116.9, 113.9, 88.3, ​​73.8, 69.8, 68.9, 66.2, 49.8, 46.0, 42.5, 40.5, 39.2, 36.6, 33.9, 30.4, 29.4, 28.7, 26.7, 24.5, 23.6.

[0095] Example 7: 4-(((1r,4r)-4-(2-(3-fluoro-4-(trifluoromethoxy)phenyl)acetamido)cyclohexyl)oxy)benzoic acid

[0096] (1r,4r)-4-hydroxycyclohexyl)tert-butyl carbamate (3.0 g, 13.95 mmol) and 10 mL of dry DMF were added to a single-necked flask. The mixture was cooled to below 0 °C, and NaH (0.62 g, 15.5 mmol) was added. After reacting at room temperature for 0.5 h, 4-fluorobenzonitrile (1.69 g, 13.95 mmol) was added. The reaction was monitored by TLC after 6 h to indicate completion. The reaction solution was poured into 200 mL of ice water, resulting in the precipitation of a white solid. The solid was filtered and dried to obtain ((1r,4r)-4-(4-cyanophenoxy)cyclohexyl)tert-butyl carbamate (intermediate 8), with a yield of 3.61 g and a yield of 82%.

[0097] Intermediate 8 (3.61 g, 11.42 mmol), CF3COOH (3 mL), and DCM (10 mL) were added to a single-necked flask and stirred at room temperature for 6 h. The reaction was monitored by TLC until it was complete. The solvent was removed under reduced pressure, and the product was not purified and directly added to the next step.

[0098] Solid phosgene (1.69 g, 5.71 mmol) and dry DCM (50 mL) were added to a three-necked flask. The temperature was lowered to below -78 °C by cold hydrazine. A solution of 4-(((1r,4r)-4-aminocyclohexyl)oxy)benzyl nitrile, triethylamine (6.92 g, 68.52 mmol), and dry dichloromethane (60 mL) was added dropwise. After the addition was complete, the mixture was moved to room temperature and stirred for 0.5 h. The reaction was then stopped. The resulting reaction solution was concentrated to dryness under reduced pressure. Dry DCM (10 mL) was added to the residue to dissolve it, yielding an isocyanate solution for later use.

[0099] 3-fluoro-4-(trifluoromethoxy)aniline (2.22 g, 11.42 mmol), triethylamine (6.92 g, 68.52 mmol), and dry dichloromethane (50 mL) were added to a single-necked flask. The above isocyanate solution was added dropwise, and the reaction was allowed to proceed at room temperature for 0.5 h. TLC showed that the reaction was complete. The reaction solution was poured into water (40 mL), extracted with EA (40 mL × 3), and then washed successively with 1 mol / L HCl (40 mL × 2), water (40 mL × 2), saturated brine (40 mL), and dried over anhydrous sodium sulfate. The solution was filtered, and the filtrate was concentrated under reduced pressure to obtain 1-((1r,4r)-4-(4-cyanophenoxy)cyclohexyl)-3-(3-fluoro-4-(trifluoromethoxy)phenyl)urea (intermediate 10), which was not purified and directly added to the next step.

[0100] Add the intermediate 10 obtained in the previous step, dissolved in tetrahydrofuran (30 mL), sodium hydroxide (0.55 g, 13.70 mmol), and water (10 mL) to a single-necked flask, and reflux. After 12 h, TLC showed that the reaction was complete. The reaction solution was concentrated under reduced pressure to remove tetrahydrofuran. The residue was added to water (300 mL), placed in a cold trap, and the pH was adjusted to 3 with 6N hydrochloric acid (10 mL). A pale yellow solid precipitated. The solid was filtered, and the filter cake was washed with water (20 mL) to obtain 4-(((1r,4r)-4-(2-(3-fluoro-4-(trifluoromethoxy)phenyl)acetamido)cyclohexyl)oxy)benzoic acid (intermediate 11). The yield was 4.06 g, with a yield of 78.0%.

[0101] Example 8: Methyl benzoate ((R)-12-((S)-7,8-difluoro-6,11-dihydrodibenzo[b,e]thiophene-11-yl)-6,8-dioxo-3,4,6,8,12-12a-hexahydro-1H-[1,4]oxazin[3,4-c]pyrido[2,1-f][1,2,4]triazin-7-yl)oxy)benzoate (ATV-123)

[0102] According to the method described in Example 1, using 4-(((1r,4r)-4-(2-(3-fluoro-4-(trifluoromethoxy)phenyl)acetamido)cyclohexyl)oxy)benzoic acid (intermediate 11) as a starting material, 0.59 g of compound ATV-123 was synthesized as a white solid, with an overall yield of 54.7% in both steps. 1H NMR(600MHz,DMSO-d6)δ8.71(s,1H),7.91-7.85(m,2H),7.66(dd,J=13.4,2.5Hz,1H),7.38-7.35(m,2H),7.21(d,J=7.8Hz,1H),7.16-7.12(m,1H),7.10-7.04(m,4H),7.01(dd,J=8.0,1.5Hz,1H),6.86-6.82(m,1H),6.30(d,J=7.6Hz,1H),5.98(d,J=6.5Hz,1H),5.93(d,J=6.6Hz,1H),5.73-5.68(m,2H),5.39(dd,J=14.4,2.4Hz,1H),4.50-4.43(m,1H),4.39(dd,J=9.9,3.1Hz,1H),4.26-4.18(m,1H),4.04(d,J=14.3Hz,1H),3.95(dd,J=10.9,3.1Hz,1H),3.57-3.44(m,2H),3.38(t,J=10.4Hz,1H),3.31(s,1H),2.96-2.90(1H),2.86-2.80(m,1H),2.08-2.01(m,2H),1.95-1.88(m,2H),1.53-1.44(m,2H),1.41-1.33(m,2H). 13 C NMR(151MHz,DMSO-d6)δ173.6,165.3,162.1,155.0,154.6,154.5,153.3,151.9,151.8,150.4,150.3,150.2,147.9,146.4,146.3,141.9,141.8,141.2,136.4,134.2,132.7,132.1,130.2,129.4,128.8,128.8,128.6,127.8,127.0,127.0,125.5,125.1,125.0,124.7,123.2,121.8,121.5,119.8,117.0,116.9,115.7,114.0,114.0,114.0,106.1,106.0,88.7,74.8,73.7,69.8,68.8,65.9,60.2,47.7,46.0,40.5,30.3,30.0,23.6,21.2,14.5.

[0103] Example 9: (((R)-12-((S)-7,8-difluoro-6,11-dihydrodibenzo[b,e]thiophene-11-yl)-6,8-dioxo-3,4,6,8,12,12a-hexahydro-1H-[1,4-c]oxazino[3,4-c]pyrido[2,1-f][1,2,4]triazin-7-yl)oxy)methyl4-(((1r,4r)-4-(3-(4-(trifluoromethoxy))ATV-126)

[0104] According to the method described in Example 1, using t-TUCB as the starting material, 0.43 g of compound ATV-126 was synthesized as a white solid, with an overall yield of 46.7% in both steps. 1 H NMR (600MHz, DMSO-d6) δ8.51 (s, 1H), 7.90 (d, J=8.8Hz, 2H), 7.50-7.45 (m, 2H) , 7.39 (dd, J=8.4, 5.1Hz, 2H), 7.22 (dd, J=8.1, 2.8Hz, 3H), 7.18-7.14 (m, 1H), 7 .12-7.06 (m, 3H), 7.02 (dd, J=7.9, 1.5Hz, 1H), 6.88-6.83 (m, 1H), 6.20 (d, J=7. 6Hz, 1H), 6.00 (d, J=6.6Hz, 1H), 5.95 (d, J=6.6Hz, 1H), 5.76-5.69 (m, 2H), 5.41 (dd, J=14.4, 2.4Hz, 1H), 4.52-4.45 (m, 1H), 4.41 (dd, J=9.9, 3.1Hz, 1H), 4.27 -4.20 (m, 1H), 4.06 (d, J=14.3Hz, 1H), 3.97 (dd, J=10.9, 3.1Hz, 1H), 3.58-3.47 (m, 2H), 3.40 (t, J=10.4Hz, 1H), 2.98-2.91 (m, 1H), 2.88-2.81 (m, 1H), 2.10-2. 03 (m, 2H), 1.94 (dd, J=13.1, 3.9Hz, 2H), 1.55-1.45 (m, 2H), 1.43-1.32 (m, 2H). 13C NMR (151MHz, DMSO-d6) δ173.6, 165.3, 162.1, 154.9, 154.5, 150.4, 150.3, 146.4, 146. 3, 142.4, 141.2, 140.3, 136.4, 134.2, 132.7, 132.7, 132.1, 130.2, 129.4, 128.6, 127. 8, 127.0, 125.5, 125.1, 125.1, 122.1, 121.8, 121.5, 119.8, 119.0, 117.0, 116.9, 115.7, 114.0, 88.7, 74.9, 73.7, 69.8, 68.8, 65.9, 47.6, 46.0, 40.5, 30.4, 30.0, 23.6, 14.6.

[0105] Example 10: 12-({[(1,3-dimethyltricyclo[3.3.1.1)] 3，7 [decyl-7-yl)amino]carbonyl}amino)dodecanoic acid-{[(12aR)-12-[(11S)-7,8-difluoro-6,11-dihydrodibenzo[1,2-e:1′,2′-b]thiacycloheptan-11-yl]-6,8-dioxane-1,3,4,6,12,12a-hexahydro[1,4]oxazacyclohexane[3,4-c]pyrido[2,1-f][1,2,4]triazacyclohexane-7-yl]oxy}methyl ester (ATV-140)

[0106] According to the methods described in Examples 1 and 5, 12-({[(1,3-dimethyltricyclo[3.3.1.1)) 3.7 Using decanoic acid (decyl-7-yl)amino[carbonyl]amino)dodecanoic acid as a starting material, compound ATV-140 was synthesized in a total of 0.38 g as a white solid, with an overall yield of 41.9% in both steps. 1H NMR (600MHz, DMSO-d6) δ7.41 (dd, J=8.2, 5.6Hz, 2H), 7.22 (d, J=7.8Hz, 1H), 7.18-7.12 (m, 1H), 7.12-7.07 (m, 1H), 7.04-7.00 (m, 1H), 6.89-6.83 (m, 1H), 7.30-5.66 (m, 4H), 5.57 (t, J = 5.6Hz, 1H), 5.43 (d, J = 12.9Hz, 2H), 4.47-4.38 (m, 2H), 4.09-3.97 (m , 2H), 3.67 (dd, J=11.6, 3.4Hz, 1H), 3.48 (t, J=10.4Hz, 1H), 3.31-3.26 (m, 1H), 2.97-2.91 (m, 1H), 2.89 (q, J=6.5Hz, 2H), 2.31 (t, J=7.4Hz, 2H), 2.04-2.01 (m, 1H), 1.67 (d, J=3.2Hz, 2H), 1.57-1.45 (m, 6H), 1.33-1.19 (m, 20H), 1.07 (s, 2H), 0.79 (s, 6H). 13 C NMR (151MHz, DMSO-d6) δ173.7, 172.9, 157.6, 154.6, 150.6, 150.2, 141.1, 136.4, 134.3, 132.7, 130.2, 129.3, 128.6, 127.8, 127.0, 125.4, 125.2, 125.1, 117.0, 116.9, 113.9, 88 .3, 73.7, 69.8, 68.9, 66.2, 60.2, 51.4, 50.8, 48.6, 46.0, 42.9, 41.0, 40.5, 39.2, 34.0, 32.4, 32.3, 30.6, 30.5, 30.1, 29.5, 29.4, 29.4, 29.3, 28.9, 26.9, 24.5, 23.6, 21.2, 14.6.

[0107] Example 11: 12-{[({4-[(trifluoromethyl)oxy]phenyl}amino)carbonyl]amino}dodecanoic acid-{[(12aR)-12-[(11S)-7,8-difluoro-6,11-dihydrodibenzo[1,2-e:1′,2′-b]thiacycloheptan-11-yl]-6,8-dioxane-1,3,4,6,12,12a-hexahydro[1,4]oxazacyclohexane[3,4-c]pyrido[2,1-f][1,2,4]triazacyclohexane-7-yl]oxy}methyl ester (ATV-148)

[0108] According to the methods described in Examples 1 and 5, 0.45 g of compound ATV-148 was synthesized as a white solid from 12-{[({4-[(trifluoromethyl)oxy]phenyl}amino)carbonyl]amino}dodecanoic acid, with an overall yield of 39.9% for both steps. 1 H NMR (600MHz, DMSO-d6) δ8.59 (s, 1H), 7.50-7.46 (m, 2H), 7.40 (d, J=6.8Hz, 2H), 7.21 (t, J=8.0Hz, 3H), 7.17-7.13 (m, 1H), 7.09 (d, J=8.0Hz, 1H), 7.04-7.00 (m, 1H), 6.15 (t, J=5.7Hz, 1H), 5.71 (dd, J=9.7, 6.1Hz, 4H), 5.42 (dd, J=14.3, 2.3Hz, 1H), 4.47-4.39 (m, 2H), 4.05 (d, J=14.3Hz, 1H), 3.99 (dd, J=10.9, 3.0Hz, 1H), 3.67 (dd, J=11.6, 3.3Hz, 1H), 3.47 (t, J=10.4Hz, 1H), 3.31-3.25 (m, 1H ), 3.06 (q, J=6.5Hz, 2H), 2.96-2.90 (m, 1H), 2.31 (t, J=7.4Hz, 2H), 1.55-1.50 (m, 2H), 1.42-1.37 (m, 2H), 1.25 (d, J=5.4Hz, 15H). 13 C NMR(151 MHz, DMSO-d6) δ173.7, 172.9, 155.5, 154.6, 152.0, 151.9, 150.6, 150.3, 150.2, 148.0, 147.9, 146.4, 14 6.3, 142.4, 141.1, 140.4, 136.4, 134.2, 132.7, 132.7, 130.2, 129.3, 128.5, 127.8, 127.0, 125.4, 125.2 , 125.1, 123.2, 122.0, 121.5, 119.8, 119.0, 117.0, 116.9, 113.9, 88.3, ​​73.7, 69.8, 68.9, 66.2, 46.0, 40.5, 39.5, 34.7, 34.0, 30.2, 29.5, 29.4, 29.4, 29.2, 28.9, 28.9, 26.8, 24.5, 23.6, 23.5, 22.9, 19.7, 11.7.

[0109] Example 12: 12-{[({3-fluoro-4-[(trifluoromethyl)oxy]phenyl}amino)carbonyl]amino}dodecanoic acid-{[(12aR)-12-[(11S)-7,8-difluoro-6,11-dihydrodibenzo[1,2-e:1′,2′-b]thiacycloheptan-11-yl]-6,8-dioxane-1,3,4,6,12,12a-hexahydro[1,4]oxazacyclohexane[3,4-c]pyrido[2,1-f][1,2,4]triazacyclohexane-7-yl]oxy}methyl ester (ATV-149)

[0110] According to the methods described in Examples 1 and 5, 0.47 g of compound ATV-149 was synthesized as a white solid from 12-{[({3-fluoro-4-[(trifluoromethyl)oxy]phenyl}amino)carbonyl]amino}dodecanoic acid, with an overall yield of 45.9% for both steps. 1 H NMR (600MHz, DMSO-d6) δ8.82 (s, 1H), 7.68 (dd, J=13.5, 2.5Hz, 1H), 7.42-7.35 (m, 3H), 7.21 (d, J=7.7Hz, 1H), 7.16-7.12 (m, 1H), 7.12-7.08 (m, 2H), 7.02-7.00 (m, 1H), 6.86-6.83 (m, 1H), 6.27 (t, J=5.7Hz, 1H), 5.72-5.68 (m, 4H), 5.42 (dd, J=14.4, 2.3Hz, 1H) , 4.47-4.38 (m, 2H), 4.05 (d, J=14.3Hz, 1H), 3.99-3.96 (m, 1H), 3.68-3.64 (m, 1H), 3.47 (t, J=10.4Hz, 1H), 3.30-3.25 (m, 1H), 3. 06 (q, J=6.6Hz, 2H), 2.96-2.90 (m, 3.5Hz, 1H), 2.30 (t, J=7.4Hz, 2H), 1.55-1.49 (m, 2H), 1.42-1.37 (m, 2H), 1.29-1.22 (m, 14H). 13C NMR(151 MHz, DMSO-d6) δ173.7, 172.9, 155.2, 154.9, 154.6, 153.3, 150.6, 150.3, 150.2, 146.4, 146.3, 142.1, 142.0, 141.1, 1 36.4, 134.2, 132.7, 130.2, 129.3, 128.8, 128.7, 128.5, 127.8, 127.0, 125.4, 125.2, 125.1, 124.6, 121.5, 119.8, 117. 0, 116.9, 114.0, 114.0, 113.9, 106.1, 105.9, 88.3, ​​73.7, 69.8, 68.9, 66.2, 46.0, 41.3, 40.5, 39.5, 35.9, 34.7, 34.4, 34.0, 30.1, 29.5, 29.4, 29.4, 29.2, 29.2, 28.9, 28.9, 27.6, 26.8, 25.2, 24.5, 23.6, 22.9, 21.1, 20.3, 19.7, 19.1, 11.7.

[0111] Example 13: 11-{[(three rings[3.3.1.1 3，7 [dec-1-ylamino]carbonyl]amino}undecanoic acid-{[(12aR)-12-[(11S)-7,8-difluoro-6,11-dihydrodibenzo[1,2-e:1′,2′-b]thiacycloheptan-11-yl]-6,8-dioxane-1,3,4,6,12,12a-hexahydro[1,4]oxazacyclohexane[3,4-c]pyrido[2,1-f][1,2,4]triazacyclohexane-7-yl]oxy}methyl ester (ATV-150)

[0112] According to the methods described in Examples 1 and 5, with 11-{[(three rings[3.3.1.1 3，7 Using [dec-1-ylamino]carbonyl]amino}undecanoic acid as a starting material, compound ATV-150 was synthesized in a total of 0.45 g as a white solid, with an overall yield of 49.9% in both steps. 1H NMR (600MHz, DMSO-d6) δ7.43-7.38 (m, 2H), 7.21 (d, J=7.7Hz, 1H), 7.15 (t, J=7.5Hz, 1H), 7.09 (d, J=7.9Hz, 1H), 7.02 (d, J=7.7Hz, 1H ), 6.85 (t, J=7.4Hz, 1H), 5.73-5.67 (m, 4H), 5.57 (t, J=5.6Hz, 1H), 5.41 (s, 1H), 4.46-4.38 (m, 2H), 4.06 (d, J=14.4Hz, 1H), 3.99 (dd , J=10.9, 3.0Hz, 1H), 3.67 (dd, J=11.6, 3.3Hz, 1H), 3.47 (t, J=10.4Hz, 1H), 3.30-3.25 (m, 1H), 2.96-2.91 (m, 1H), 2.89 (q, J=6.5Hz, 2H), 2.31 (t, J=7.5Hz, 2H), 1.99-1.95 (m, 3H), 1.83 (d, J=2.9Hz, 6H), 1.62-1.55 (m, 6H), 1.53 (q, J=7.3Hz, 2H), 1.33-1.18 (m, 15H). 13 C NMR (151MHz, DMSO-d6) δ173.7, 172.9, 157.5, 154.6, 150.6, 150.3, 150.2, 147.9, 146.4, 141.1, 136.4, 134.2, 132.7, 130.2, 129.3, 128.6, 127.8, 127.0, 125.4, 125 .2, 125.1, 117.0, 116.9, 113.9, 88.3, ​​73.7, 69.8, 68.9, 66.2, 49.8, 46.0, 42.5, 40.5, 39.2, 36.6, 34.0, 30.5, 29.5, 29.4, 29.3, 29.3, 29.2, 28.9, 26.9, 24.5, 23.6.

[0113] Example 14: 11-({[(3,5-dimethyltricyclo[3.3.1.1 3，7 [dec-1-yl)amino]carbonyl}amino)undecanoic acid-{[(12aR)-12-[(11S)-7,8-difluoro-6,11-dihydrodibenzo[1,2-e:1′,2'-b]thiacycloheptan-11-yl]-6,8-dioxane-1,3,4,6,12,12a-hexahydro[1,4]oxazacyclohexane[3,4-c]pyrido[2,1-f][1,2,4]triazacyclohexane-7-yl]oxy}methyl ester (ATV-151)

[0114] According to the methods described in Examples 1 and 5, 11-({[(3,5-dimethyltricyclo[3.3.1.1)) 3，7 Using dec-1-yl)amino]carbonyl}amino)undecanoic acid as a starting material, compound ATV-151 was synthesized in a total of 0.46 g of white solid, with an overall yield of 41.3% in both steps. 1 H NMR (600MHz, DMSO-d6) δ7.40 (dd, J=8.3, 4.7Hz, 2H), 7.21 (d, J=7.8Hz, 1H), 7.16-7.13 (m, 1H), 7.09 (dd, J=8.0, 1.3Hz, 1H), 7.02 (dd, J=7.9, 1 .5Hz, 1H), 6.86-6.83 (m, 1H), 5.73-5.68 (m, 4H), 5.57 (t, J=5.6Hz, 1H), 5.43 (d, J=12.0Hz, 2H), 4.46-4.39 (m, 2H), 4.06 (d, J=14.3Hz, 1H), 3. 99 (dd, J=10.8, 3.1Hz, 1H), 3.67 (dd, J=11.4, 3.3Hz, 1H), 3.47 (t, J=10.4Hz, 1H), 3.31-3.25 (m, 1H), 2.96-2.91 (m, 1H), 2.89 (q, J=6.5Hz, 2H) , 2.33-2.29 (m, 2H), 2.04-2.01 (m, 1H), 1.66 (d, J=3.1Hz, 2H), 1.54 (q, J=7.0Hz, 2H), 1.48 (d, J=5.0Hz, 4H), 1.33-1.18 (m, 20H), 0.79 (s, 6H). 13C NMR(151 MHz, DMSO-d6) δ173.7, 172.9, 157.6, 154.6, 152.0, 151.9, 150.6, 150.3, 150.2, 148.0, 147.9, 146.4, 146.3, 141.1 , 136.4, 134.2, 132.7, 130.2, 129.3, 128.6, 127.8, 127.0, 125.4, 125.2, 125.1, 117.0, 116.9, 113.9, 88.3, ​​73.7, 69 .8, 68.9, 66.2, 51.4, 50.8, 48.6, 46.0, 42.9, 41.3, 41.0, 40.5, 39.2, 35.9, 34.7, 34.4, 34.0, 32.3, 30.6, 30.5, 30.1, 29.5, 29.3, 29.3, 29.0, 28.9, 27.6, 26.9, 25.3, 24.5, 23.6, 23.6, 23.0, 21.1, 20.4, 19.7, 19.1, 14.6, 14.4, 11.7.

[0115] Example 15: 8-{[(three rings[3.3.1.1 3，7 [dec-3-ylamino]carbonyl]amino}octanoic acid-{[(12aR)-12-[(11S)-7,8-difluoro-6,11-dihydrodibenzo[1,2-e:1′,2′-b]thiacycloheptam-11-yl]-6,8-dioxane-1,3,4,6,12,12a-hexahydro[1,4]oxazacyclohexam[3,4-c]pyrido[2,1-f][1,2,4]triazacyclohexam-7-yl]oxy}methyl ester (ATV-157)

[0116] According to the methods described in Examples 1 and 5, with 8-{[(three rings[3.3.1.1 3，7 Using dec-3-ylamino)carbonyl]amino}octanoic acid as a starting material, compound ATV-157 was synthesized as a white solid (0.36 g), with an overall yield of 47.3% in both steps. 11H NMR (600 MHz, DMSO-d6) δ 7.41 (dd, J = 9.5, 6.3 Hz, 2H), 7.22 (d, J = 7.7 Hz, 1H), 7.16 (t, J = 7.6 Hz, 1H), 7.09 (d, J = 8.0 Hz, 1H), 7.02 (d, J = 7.7 Hz, 1H), 6.85 (t, J = 7.5 Hz, 1H), 5.73 - 5.66 (m, 4H), 5.59 (t, J = 5.6 Hz, 1H), 5.43 (d, J = 17.1 Hz, 2H), 4.47 - 4.38 (m, 2H), 4.06 (d, J = 14.4 Hz, 1H), 3.99 (dd, J = 10.9, 3.1 Hz, 1H), 3.68 (dd, J = 11.5, 3.3 Hz, 1H), 3.47 (t, J = 10.4 Hz, 1H), 3.31 - 3.25 (m, 1H), 2.98 - 2.87 (m, 3H), 2.31 (t, J = 7.5 Hz, 2H), 1.97 (s, 3H), 1.83 (d, J = 3.0 Hz, 6H), 1.61 - 1.56 (m, 6H), 1.56 - 1.52 (m, 2H), 1.33 - 1.27 (m, 4H), 1.24 (q, J = 9.7 Hz, 4H). 13 13C NMR (151 MHz, DMSO-d6) δ 173.7, 172.9, 157.5, 154.6, 152.0, 151.9, 150.6, 150.2, 148.0, 147.9, 146.4, 146.3, 141.1, 136.4, 134.2, 132.7, 132.7, 130.2, 129.3, 128.6, 127.8, 127.0, 127.0, 125.4, 125.2, 125.1, 117.1, 116.9, 113.9, 88.2, 73.7, 69.8, 68.9, 66.2, 49.8, 46.0, 42.5, 42.1, 40.5, 39.2, 36.6, 34.7, 33.9, 33.6, 30.5, 29.5, 29.4, 29.0, 28.9, 28.7, 26.8, 25.3, 24.4, 23.6.

[0117] Example 16: 12-[({[1-(Tricyclo[3.3.1.1 3，7[dec-1-yl]ethyl]amino]carbonyl]amino]dodecanoic acid-{[(12aR)-12-[(11S)-7,8-difluoro-6,11-dihydrodibenzo[1,2-e:1′,2′-b]thiacycloheptan-11-yl]-6,8-dioxane-1,3,4,6,12,12a-hexahydro[1,4]oxazacyclohexane[3,4-c]pyrido[2,1-f][1,2,4]triazacyclohexane-7-yl]oxy}methyl ester (ATV-158)

[0118] According to the methods described in Examples 1 and 5, with 12-[({[1-(three rings[3.3.1.1 3，7 Using dec-1-yl)ethyl]amino]carbonyl]amino]dodecanoic acid as a starting material, compound ATV-158 was synthesized as a white solid (0.41 g), with an overall yield of 44.3% in both steps. 1 H NMR (600MHz, DMSO-d6) δ7.40 (t, J=6.5Hz, 2H), 7.22 (d, J=7.7Hz, 1H), 7.15 (t, J=7.7Hz, 1H), 7.09 (d, J=7.9Hz, 1H), 7.02 (d, J=7.7Hz, 1H), 6.85 (t, J=7.5Hz, 1H), 5.73-5.68 (m, 4H), 5.65 (t, J=5.6Hz, 1H), 5.50 (d, J=9.5Hz, 1H), 5.43 (dd, J=14.1, 2.4Hz, 1H), 4.47-4.38 (m, 2H), 4.06 (d, J=1 4.3Hz, 1H), 3.99 (dd, J=11.0, 3.1Hz, 1H), 3.67 (dd, J=11.6, 3.3Hz, 1H), 3.47 (t, J=10.4Hz, 1H), 3.31-3.24 (m, 2H), 2.98-2.90 (m, 3H), 2.31 (t, J =7.4Hz, 2H), 1.94-1.89 (m, 3H), 1.64 (d, J = 12.0Hz, 3H), 1.61-1.50 (m, 6 H), 1.46 (d, J=12.4Hz, 4H), 1.41 (d, J=12.3Hz, 3H), 1.32-1.19 (m, 17H). 13C NMR (151MHz, DMSO-d6) δ173.7, 172.9, 158.3, 154.6, 152.0, 151.9, 150.6, 150.3, 150.2, 148.0, 147.9, 146.4, 146. 3, 141.1, 136.4, 134.2, 132.7, 132.7, 130.2, 129.3, 128.5, 127.8, 127.0, 127.0, 125.4, 125.2, 125.1, 117.0, 116. 9, 113.8, 88.3, ​​73.7, 69.8, 68.9, 66.2, 53.0, 46.0, 41.3, 40.5, 38.5, 37.2, 36.1, 35.9, 34.7, 34.0, 30.5, 29.5, 29.4, 29.4, 29.3, 29.3, 29.0, 28.9, 28.3, 26.9, 25.3, 24.5, 23.6, 23.6, 22.9, 21.1, 20.4, 19.7, 19.1, 15.6, 14.6, 11.7.

[0119] Example 17: (((R)-12-((S)-7,8-difluoro-6,11-dihydrodibenzo[b,e]thiophene-11-yl)-6,8-dioxo-3,4,6,8,12,12a-hexahydro-1H-[1,4-c]oxazino[3,4-c]pyrido[2,1-f][1,2,4]triazin-7-yl)oxy)methyl12-(3-(((3r,5r,7r)-adamantane (ATV-159))

[0120] According to the methods described in Examples 1 and 5, with 12-[({[1-(three rings[3.3.1.1 3，7 Using dec-1-yl)ethyl]amino]carbonyl]amino]dodecanoic acid as a starting material, compound ATV-159 was synthesized as a white solid (0.41 g), with an overall yield of 44.3% in both steps. 1H NMR(600MHz,DMSO-d6)δ7.40(t,J=6.6Hz,2H),7.22(d,J=7.7Hz,1H),7.15(t,J=7.6Hz,1H),7.09(d,J=7.9Hz,1H),7.02(d,J=7.7Hz,1H),6.85(t,J=7.5Hz,1H),5.73-5.65(m,6H),5.43(dd,J=14.4,2.4Hz,1H),4.47-4.38(m,2H),4.06(d,J=14.3Hz,1H),3.99(dd,J=10.9,3.1Hz,1H),3.67(dd,J=11.6,3.4Hz,1H),3.47(t,J=10.4Hz,1H),3.31-3.25(m,1H),2.98-2.90(m,3H),2.68(d,J=6.1Hz,2H),2.31(t,J=7.4Hz,2H),1.93-1.88(m,3H),1.65(d,J=12.1Hz,3H),1.59-1.50(m,6H),1.39(d,J=2.8Hz,6H),1.29-1.20(m,16H). 13 C NMR(151MHz,DMSO-d6)δ173.7,172.9,158.8,154.6,150.6,150.2,147.9,146.4,146.3,141.1,136.4,134.2,132.7,130.2,129.3,128.5,127.8,127.0,127.0,125.4,125.2,125.1,117.0,116.9,113.8,88.3,73.7,69.8,68.9,66.2,51.5,46.0,39.6,37.1,35.9,34.7,34.0,34.0,33.7,30.5,29.5,29.4,29.4,29.3,29.3,29.1,29.0,28.9,28.3,28.2,26.9,25.3,24.7,24.5,23.6,22.9,19.1,14.6,11.7.

[0121] Example 18: 12-({[(bicyclo[2.2.1]hept-2-en-6-ylmethyl)amino]carbonyl}amino)dodecanoic acid-{[(12aR)-12-[(11S)-7,8-difluoro-6,11-dihydrodibenzo[1,2-e:1′,2′-b]thiocycloheptan-11-yl]-6,8-dioxane-1,3,4,6,12,12a-hexahydro[1,4]oxazacyclohexane[3,4-c]pyrido[2,1-f][1,2,4]triazacyclohexane-7-yl]oxy}methyl ester (ATV-160)

[0122] According to the method described in Example 1, 0.49 g of compound ATV-160 was synthesized from 12-({[(bicyclo[2.2.1]hept-2-en-6-ylmethyl)amino]carbonyl}amino)dodecanoic acid as a raw material, with an overall yield of 47.1% for both steps. 1 H NMR (600MHz, DMSO-d6) δ7.40 (d, J=6.4Hz, 2H), 7.22 (d, J=7.8Hz, 1H), 7.15 (t, J=7.6Hz, 1H), 7.09 (d, J=7.9Hz, 1H), 7.04-7.00 (m, 1H), 6. 87-6.83 (m, 1H), 6.13 (dd, J=5.8, 3.0Hz, 1H[), 5.93 (dd, J=5.8, 2.8Hz, 1H), 5.77-5.66 (m, 6H), 5.43 (dd, J=14.4, 2.4Hz, 1H), 4.47-4.38 ( m, 2H), 4.06 (d, J=14.3Hz, 1H), 3.99 (dd, J=10.9, 3.0Hz, 1H), 3.67 (dd, J=11.5, 3.3Hz, 1H), 3.47 (t, J=10.4Hz, 1H), 3.31-3.25 (m, 1H), 2. 94-2.90 (m, 3H), 2.80-2.72 (m, 3H), 2.31 (t, J=7.4Hz, 2H), 2.13-2.06 (m, 1H), 1.76-1.70 (m, 1H), 1.55-1.49 (m, 2H), 1.36-1.17 (m, 20H). 13C NMR(151 MHz, DMSO-d6) δ173.7, 172.9, 158.5, 158.4, 154.6, 152.0, 151.9, 150.6, 150.3, 150.2, 148.0, 147.9, 146.4, 146.3, 141.1, 137 .3, 136.9, 136.9, 136.4, 134.2, 132.8, 132.7, 132.7, 130.2, 129.3, 128.5, 127.8, 127.0, 125.4, 125.2, 125.1, 117.0, 116.9, 11 3.9, 88.3, ​​73.7, 69.8, 68.9, 66.2, 49.4, 46.0, 45.1, 44.9, 44.1, 43.9, 42.3, 41.6, 41.3, 40.5, 39.6, 35.9, 34.7, 34.4, 34.0, 30.8, 30.5, 30.2, 29.5, 29.4, 29.4, 29.3, 29.3, 29.0, 28.9, 26.9, 25.3, 24.5, 23.6, 23.0, 21.1, 20.4, 19.7, 19.1, 14.6, 14.3, 11.7.

[0123] Example 19: In vitro bioactivity and cytotoxicity studies

[0124] Test compound: The compound of this invention and the control compound baloxavir. In vitro bioactivity study method: MDCK cells were seeded at a density of 20,000 cells per well in 96-well cell culture plates and then incubated overnight at 37°C in a 5% CO2 incubator. The next day, influenza virus A / WSN / 1933 (H1N1) strain was introduced at 2*TCID50 per well. 90 The compound was added to cell culture wells after dilution (2-fold serial dilution, 8 test concentration points), with a final DMSO concentration of 0.5% in the culture medium. Cell plates were incubated at 37°C in a 5% CO2 incubator for 2 days. Cell viability was assessed using a CCK8 cell viability assay kit after 2 days of culture. The raw data were analyzed using GraphPadPrism 9.0 software with nonlinear fitting to determine the EC50 concentration. 50 The values ​​are shown in Table 1.

[0125] Methodology for cytotoxicity studies: Cytotoxicity assays and antiviral activity assays were performed in parallel, with all experimental conditions identical to the antiviral activity assays except for the absence of virus. Cell viability was assessed using a CCK8 cell viability assay kit after 2 days of culture. Raw data were used for compound cytotoxicity (CCK) studies. 50 The calculations are shown in Table 1.

[0126] Table 1. Inhibitory activity and toxicity of the compounds against influenza virus A / WSN / 1933 (H1N1)

[0127] As shown in Table 1, BAX and MBAX have ECGs against H1N1. 50 The values ​​were 2.19 nM and 1.68 nM, respectively, very close to those previously reported in the literature. In contrast, most of the twin drugs showed strong inhibitory effects against H1N1. Specifically, ATV-114, ATV-148, ATV-149, ATV-150, ATV-151, and ATV-158 showed better antiviral activity against influenza than MBAX, especially ATV-114 against the EC50 of H1N1. 50 The concentration was 0.51 nM. ATV-114 was synthesized by linking the carboxyl structure of the sEH inhibitor AUDA to the enol structure of BAX via a methylene group. Therefore, the inventors evaluated the inhibitory activity of compounds ATV-114 and MBAX against various clinical isolates of influenza A and influenza B viruses, including the oseltamivir-resistant strain A / PR / 8 / 1934 (NA-H274Y). Notably, ATV-114 exhibited superior inhibitory activity against all tested strains compared to MBAX. Particularly against the A / PR / 8 / 1934 (H1N1) virus, ATV-114 showed almost four times the activity of MBAX, as shown in Table 2.

[0128] Table 2 Inhibitory activities of ATV-114 and MBAX against different influenza virus strains

[0129] Example 20: ATV-114 is more effective than MBAX in inhibiting viral proteins in two cell types with high and low AADAC enzyme expression.

[0130] To exert its antiviral activity, maloxavir must be converted to baloxaviric acid, a process dependent on arylacetamide deacetylase (AADAC). AADAC is primarily expressed in the liver, intestine, pancreas, and adrenal glands, with low levels detected in other tissues. In contrast, carboxylhydrolases that hydrolyze ATV-114 into the active metabolites AUDA and BAX are widely distributed throughout the body. In the experiments of this invention, cells were infected with H1N1 virus and treated with MBAX, ATV-114, and AUDA under conditions of high and low AADAC expression. As shown in Figure 1, AF represents the inhibitory effect of different concentrations of MBAX, ATV-114, and AUDA on H1N1 viral protein expression in cell lines with high AADAC expression (i.e., Caco-2, HEPG-2, and MDCK cells); DF represents the inhibitory effect of different concentrations of MBAX, ATV-114, and AUDA on H1N1 viral protein expression in cell lines with low AADAC expression (A549, Beas-2B, and HaCat cells).

[0131] Figure 1 shows that MBAX significantly inhibited the expression of influenza virus nucleoprotein (NP) and polymerase acid (PA) proteins, with enhanced efficacy observed in cells exhibiting high AADAC expression. In cells with high AADAC expression, 10 nM MBAX almost completely inhibited influenza virus protein expression, while ATV-114 showed more significant inhibitory effects on viral protein expression in both high and low AADAC expression cells, achieving complete inhibition of viral protein expression at a concentration of 5 nM.

[0132] Example 21: ATV-114 exerts anti-inflammatory activity by inhibiting NF-κB signaling.

[0133] In this study, the inventors used an LPS-induced RAW264.7 inflammation model to evaluate the anti-inflammatory effect of AUDA in vitro and determined that ATV-114 could successfully hydrolyze AUDA to obtain a similar anti-inflammatory effect. As shown in Figure 2A, ATV-114 and AUDA inhibited LPS-induced upregulation of inflammatory proteins and activation of the NF-κB pathway, while MBAX had no significant effect. Subsequently, the inventors transfected RAW264.7 cells with an NF-κB luciferase reporter plasmid, and the level of luciferase expression represented NF-κB transcriptional activity. As shown in Figure 2B, the NF-κB inhibitor BAY11-7085 almost completely inhibited LPS-induced NF-κB transcriptional activation, and both AUDA and ATV-114 inhibited NF-κB transcription in a concentration-dependent manner, indicating that NF-κB is indeed a target of AUDA, and ATV-114 inhibits NF-κB by releasing AUDA molecules intracellularly.

[0134] Example 22: Confocal assay to detect the inhibitory effect of twin drugs ATV-114 on RNP

[0135] Six hours after influenza virus infection of A549 cells, the viral nucleoprotein complex RNP is released from the cell nucleus to initiate a new round of infection and replication. After the cells are infected with influenza virus, the NF-κB signaling pathway is activated, NF-κB is phosphorylated and enters the cell nucleus, promoting the nucleus exit of RNP.

[0136] Six hours after infection with A / WSN / 1933 virus, A549 cells were subjected to indirect immunofluorescence assays using NF-κB and NP antibodies. DAPI was used to indicate nuclear localization. Scale bar: 10 μm. As shown in Figure 3, compound ATV-114 inhibits RNP nuclear export by reducing NF-κB nuclear translocation.

[0137] Example 23: ATV-114 effectively protects mice from influenza virus infection and death.

[0138] The efficacy of ATV-114 in a mouse model of H1N1 influenza A virus infection was evaluated. Mice were infected with influenza A virus A / PR / 8 / 1934 or A / PR / 8 / 34 with NA-H274Y via intranasal instillation. Treatment with the compounds began at 12, 48, 72, and 96 hours post-infection. MBAX, ATV-114, and HEAUDA were administered once daily, while oseltamivir phosphate was administered once daily for three consecutive days. The anti-H1N1 effect of the compounds in this model was evaluated by observing changes in mouse body weight and survival rate, as shown in Figure 4. BALB / c mice of ATVF grade (Experimental Animal Management Center, Southern Medical University), 6-7 weeks old, with an equal number of males and females, were used in the experiment. Mice were acclimatized for at least 3 days after arrival at the BSL-2 animal facility before the experiment began. The day of infection was designated as day 0 of the experiment.

[0139] Currently, widely used antiviral drugs in clinical practice, such as oseltamivir and baloxavir, are recommended to be administered within 48 hours of infection to effectively protect influenza patients. In the inventors' research, BALB / c mice infected with A / PR / 8 / 1934 virus were orally administered MBAX or ATV-114 at time points of 48, 72, and 96 hours post-infection. ATV-114 treatment maintained a survival rate of 37.5% at 72 and 96 hours post-infection. In contrast, oseltamivir showed no protective effect when administered at 72 and 96 hours post-infection, and MBAX provided only a 12.5% ​​survival rate when administered at 72 hours post-infection. These findings highlight the significant potential of AUDA's potent anti-inflammatory effects.

[0140] In addition to the direct damage caused by the virus, influenza virus infection also poses a serious threat to lung cells due to the inflammatory cytokine storm. Therefore, although antiviral treatment initiated 72 hours after infection can suppress viral replication, high viral load coupled with an uncontrolled inflammatory response hinders survival. Due to the effects of the cytokine storm and pulmonary fibrosis, surviving mice often recover slowly, leading to irreversible lung damage.

[0141] The results indicate that ATV-114 releases AUDA during the mid-stage of viral infection to inhibit excessive activation of inflammatory pathways, thereby mitigating the effects of cytokine storm. Simultaneously, ATV-114 releases BAX, rapidly inhibiting viral replication and preventing the generation of new progeny viruses, thus reducing viral load at its source. The dual anti-inflammatory and antiviral effects of ATV-114 provide crucial therapeutic benefits to infected mice during this critical period.

[0142] The method of this invention has been described through preferred embodiments. Those skilled in the art will readily be able to modify or appropriately alter and combine the methods and applications described herein within the scope, spirit, and context of this invention to implement and apply the technology of this invention. Those skilled in the art can refer to the content herein to appropriately improve process parameters. It should be particularly noted that all similar substitutions and modifications are obvious to those skilled in the art and are considered to be included within the scope of this invention.

Claims

1. [Amended according to Rule 26, 25.02.2026] A baloxavir derivative, its stereoisomer, its crystalline hydrate, its deuterated derivative, its solvate, its prodrug, or a pharmaceutically acceptable salt thereof, characterized in that, The structural formula of the baloxavir derivative is shown in Formula I: Among them, sEH inhibitor is the esterified residue of a soluble epoxide hydrolase inhibitor.

2. The baloxavir derivative, its stereoisomer, its crystalline hydrate, its deuterated derivative, its solvate, its prodrug, or its pharmaceutically acceptable salt according to claim 1, characterized in that, The soluble epoxide hydrolase inhibitor contains a substituted benzene ring or a substituted adamantane.

3. The baloxavir derivative, its stereoisomer, its crystalline hydrate, its deuterated derivative, its solvate, its prodrug, or its pharmaceutically acceptable salt according to claim 1, characterized in that, The soluble epoxide hydrolase inhibitor is t-AUCB, 12-{[(tricyclic[3.3.1.1 3,7 [dec-1-ylamino]carbonyl]amino}dodecanoic acid (AUDA), (S)-1-(4-(3-(1r,3R,5S,7S)-3,5-dimethyladamantane-1-yl)ureo)-3-fluorobenzyl)piperidine-3-carboxylic acid, 7-{[(tricyclic[3.3.1.1 3,7 [dec-1-ylamino]carbonyl]amino}heptanoic acid, 4-(((1r,4r)-4-(2-(3-fluoro-4-(trifluoromethoxy)phenyl)acetamido)cyclohexyl)oxy)benzoic acid, t-TUCB, 12-({[(1,3-dimethyltricyclo[3.3.1.1 3,7 ]dec-7-yl)amino]carbonyl}amino)dodecanoic acid, 12-{[({4-[(trifluoromethyl)oxy]phenyl}amino)carbonyl]amino}dodecanoic acid, 12-{[({3-fluoro-4-[(trifluoromethyl)oxy]phenyl}amino)carbonyl]amino}dodecanoic acid, 11-{[(tricyclo[3.3.1.1 3,7 [dec-1-ylamino]carbonyl]amino}undecanoic acid, 11-({[(3,5-dimethyltricyclo[3.3.1.1]) 3,7 [dec-1-yl)amino]carbonyl}amino)undecanoic acid, 8-{[(tricyclic[3.3.1.1] 3,7 [dec-3-ylamino]carbonyl]amino}octanoic acid, with 12-[({[1-(tricyclic[3.3.1.1] 3,7 [dec-1-yl]ethyl]amino]carbonyl]amino]dodecanoic acid, 12-[({[1-(tricyclic[3.3.1.1] 3,7 At least one of 12-([(bicyclo[2.2.1]hept-2-en-6-ylmethyl)amino]carbonyl)amino)dodecanoic acid, decanoic acid, and 12-({[(bicyclo[2.2.1]hept-2-en-6-ylmethyl)amino]carbonyl)amino)dodecanoic acid).

4. The baloxavir derivative, its stereoisomer, its crystalline hydrate, its deuterated derivative, its solvate, its prodrug, or its pharmaceutically acceptable salt according to claim 1, characterized in that, The compound represented by Formula I is selected from the following compounds:

5. A pharmaceutical composition, characterized in that, Includes the baloxavir derivatives, stereoisomers, crystalline hydrates, deuterated derivatives, solvates, prodrugs, or pharmaceutically acceptable salts thereof as described in any one of claims 1-4.

6. The pharmaceutical composition according to claim 5, characterized in that, This includes pharmaceutically acceptable carriers.

7. The pharmaceutical composition according to claim 5, characterized in that, The pharmaceutical composition is in the form of tablets, pills, creams, emulsions, ointments, suspensions, lyophilized agents, capsules, sustained-release agents, granules, powders, injectable drugs, or sprays.

8. Use of the baloxavir derivative, its stereoisomer, its crystalline hydrate, its deuterated derivative, its solvate, its prodrug, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of any one of claims 1-4, in the preparation of anti-inflammatory and antiviral drugs.

9. The use according to claim 8, characterized in that, The virus in question is an influenza virus.

10. The use according to claim 9, characterized in that, The influenza virus in question is influenza A virus and / or influenza B virus.

11. The use according to claim 9, characterized in that, The influenza virus is at least one of H1N1, H5N1, H7N9, H3N2 and type B influenza virus.

12. The use according to claim 11, characterized in that, The influenza virus is at least one of H1N1, H3N2, or type B influenza virus.

13. Use of the baloxavir derivative, its stereoisomer, its crystalline hydrate, its deuterated derivative, its solvate, its prodrug, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of any one of claims 5-7, in the preparation of an NF-κB signaling pathway inhibitor.

14. The use according to any one of claims 8-13, characterized in that, The anti-inflammatory and antiviral drugs or the NF-κB signaling pathway inhibitors are suitable for human or animal use.

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