Pyridinone compounds having integrase inhibitory activity and pharmaceutical uses thereof

Abstract

The present disclosure provides pyridinone compounds having integrase inhibitory activity and pharmaceutical uses thereof. In particular, the present disclosure provides compounds of the structure of Formula I or pharmaceutically acceptable salts thereof, which are useful in the treatment of human immunodeficiency virus (HIV) infection.

Description

Technical Field

[0001] This disclosure pertains to the pharmaceutical field and relates to a pyridone compound with integrase inhibitory activity and its pharmaceutical uses. Background Technology

[0002] Human immunodeficiency virus (HIV), also known as human immunodeficiency virus, is the most structurally complex known retrovirus, including HIV (AIDS). In the AIDS stage, HIV infection can cause various opportunistic infections and tumors. These complications can impact a person's health and quality of life, and may even be life-threatening, thus causing widespread panic surrounding HIV.

[0003] Human immunodeficiency virus type 1 (HIV-1) encodes three enzymes required for viral replication: reverse transcriptase, protease, and integrase. Although drugs targeting reverse transcriptase and protease are widely used and have shown effectiveness, especially when used in combination, their usefulness is limited by the development of toxicity and resistant strains.

[0004] On the other hand, due to the frequent emergence of drug-resistant viruses, multidrug combination therapy is effective in the treatment of AIDS. Clinically, two of the two types of anti-HIV drugs—reverse transcriptase inhibitors and protease inhibitors—are used. However, drugs with the same mechanism of action often exhibit cross-resistance or only show additional efficacy. Therefore, there is a need to develop anti-HIV drugs with different mechanisms of action.

[0005] In this context, integrase inhibitors (WO2006116764A1 and WO2013054862A1) have attracted much attention as anti-HIV drugs with novel mechanisms of action. HIV integrase inhibitors are a type of anti-HIV / AIDS drug with a novel mechanism of action. They can be used in combination with other antiretroviral drugs to effectively treat HIV infection, and are less likely to develop drug resistance in clinical practice.

[0006] A cocktail therapy combining HIV integrase inhibitors with marketed reverse transcriptase inhibitors, protease inhibitors, and fusion inhibitors may solve the problems of drug resistance and cross-resistance associated with monotherapy, significantly improve the treatment efficacy of existing HIV drugs, and become a new treatment approach and option.

[0007] Currently approved HIV integrase inhibitors that have entered clinical use include: raltegravir, elvitegravir, dolutegravir, bicretivavir, cabotevir, and the fixed-dose combination drug Stribild. Summary of the Invention

[0008] This disclosure provides a compound of formula I or a pharmaceutically acceptable salt thereof.

[0009]

[0010] in,

[0011] R1 is selected from hydrogen, C 1-6 Alkyl, C 3-6 Cycloalkyl, 3-6 membered heterocyclic groups, hydroxyl groups; the C 1-6 Alkyl, C 3-6 Cycloalkyl groups and 3-6-membered heterocyclic groups are optionally surrounded by 1-3 R groups. 5a Replaced;

[0012] R2 is selected from hydrogen, halogen, or C with 1-3 halogen substitutions. 1-6 Alkyl, C 3-6 Cycloalkyl, 3-6 membered heterocyclic group, hydroxyl, methylenecyclopropyl, oxo; the C 3-6 Cycloalkyl, 3-6 membered heterocyclic, or methylenecyclopropyl are optionally surrounded by 1-3 R groups. 5b Replaced;

[0013] R3 is independently selected from halogen, cyano, amino, nitro, C 1-6 Alkyl, C 3-6 Cycloalkyl, 3-6 membered heterocyclic groups, hydroxyl groups; the C 1-6 Alkyl, C 3-6 Cycloalkyl groups and 3-6-membered heterocyclic groups are optionally surrounded by 1-3 R groups. 5c Replaced;

[0014] n is an integer selected from 0 to 4;

[0015] R4 is independently selected from halogen, cyano, amino, nitro, C 1-6 Alkyl, C 3-6 Cycloalkyl, 3-6 membered heterocyclic groups, hydroxyl groups, oxo groups; the C 1-6 Alkyl, C 3-6 Cycloalkyl groups and 3-6-membered heterocyclic groups are optionally surrounded by 1-3 R groups. 5d Replaced;

[0016] m is an integer between 0 and 2;

[0017] R 5a R 5b R 5c R 5d Each is independently selected from halogens, C 1-6 Alkyl, C 3-6 Cycloalkyl, 3-6 membered heterocyclic groups, hydroxyl, cyano, amino, nitro.

[0018] In some embodiments, the compound represented by Formula I can be:

[0019]

[0020] In some embodiments, in the compound represented by Formula I, Formula II or Formula III or its pharmaceutically acceptable salt, R1 is selected from hydrogen, C 1-6 Alkyl, C 3-6 cycloalkyl, hydroxyl; the C 1-6 Alkyl, C 3-6 The cycloalkyl group is optionally surrounded by 1-3 Rs. 5a Replaced;

[0021] R 5a Selected from halogens, C 1-6 Alkyl, C 3-6 Cycloalkyl, hydroxyl, cyano, amino, nitro;

[0022] R1 is preferably selected from hydrogen, C 1-6 Alkyl, C 3-6 cycloalkyl, hydroxyl; the C 1-6 Alkyl, C 3-6 The cycloalkyl group is optionally surrounded by 1-3 Rs. 5a Replaced; R 5a Selected from halogen, methyl, ethyl, isopropyl, cyclopropyl, n-propyl, hydroxy, cyano, amino, and nitro.

[0023] In some embodiments, in the compound represented by Formula I, Formula II, or Formula III, or a pharmaceutically acceptable salt thereof, R4 is independently selected from halogen, cyano, amino, nitro, C 1-6 Alkyl, C 3-6 Cycloalkyl, hydroxyl, oxo; the C 1-6 Alkyl, C 3-6 The cycloalkyl group is optionally surrounded by 1-3 Rs. 5d Replaced;

[0024] R 5d Each is independently selected from halogens, C 1-6 Alkyl, C 3-6 Cycloalkyl, hydroxyl, cyano, amino, nitro;

[0025] Preferably, R4 is independently selected from halogen, cyano, amino, nitro, C 1-6 Alkyl, C 3-6 Cycloalkyl, hydroxyl, oxo.

[0026] In some embodiments, in the compounds of Formula I, Formula II or Formula III or their pharmaceutically acceptable salts, m is an integer from 0 to 2; preferably m is an integer from 0 to 1; more preferably m is 0.

[0027] In some embodiments, in the compound represented by Formula I, Formula II, or Formula III, or a pharmaceutically acceptable salt thereof, R3 is independently selected from halogen, cyano, amino, nitro, C1-6 Alkyl, C 3-6 cycloalkyl, hydroxyl; the C 1-6 Alkyl, C 3-6 The cycloalkyl group is optionally surrounded by 1-3 Rs. 5c Replaced;

[0028] n is an integer selected from 0 to 4;

[0029] R 5c Each is independently selected from halogens, C 1-6 Alkyl, C 3-6 Cycloalkyl, hydroxyl, cyano, amino, nitro;

[0030] Preferably, R3 is independently selected from halogen, cyano, amino, nitro, C 1-6 Alkyl, C 3-6 cycloalkyl, hydroxyl; the C 1-6 Alkyl, C 3-6 The cycloalkyl group is optionally surrounded by 1-3 Rs. 5c Replaced;

[0031] n is an integer selected from 0 to 2;

[0032] R 5c Each is independently selected from halogen, methyl, ethyl, cyclopropyl, isopropyl, hydroxy, cyano, amino, and nitro;

[0033] More preferably, R3 is independently selected from halogen, cyano, amino, nitro, methyl, ethyl, cyclopropyl, isopropyl, and hydroxy; wherein the methyl, ethyl, cyclopropyl, and isopropyl groups are optionally surrounded by 1-3 R groups. 5c Replaced;

[0034] n is an integer selected from 0 to 2;

[0035] R 5c Each is independently selected from halogen, methyl, ethyl, cyclopropyl, isopropyl, hydroxy, cyano, amino, and nitro;

[0036] The optimal choice R3 is independently selected from halogens; n is selected from integers from 0 to 2.

[0037] This disclosure also provides a compound of formula IV or a pharmaceutically acceptable salt thereof.

[0038]

[0039] R1, R2, R3, R4, and m are defined as in Equation I.

[0040] This disclosure also provides a compound of formula V or a pharmaceutically acceptable salt thereof.

[0041]

[0042] R2 is defined as in Equation I.

[0043] In some embodiments, in the compounds of formula I, II, III, IV or V or their pharmaceutically acceptable salts, R2 is selected from hydrogen, halogen, or C with 1-3 halogen substitutions. 1-6 Alkyl, C 3-6 Cycloalkyl, hydroxyl, methylenecyclopropyl, oxo; the C 3-6 Cycloalkyl and methylenecyclopropyl groups are optionally surrounded by 1-3 R groups. 5b Replaced;

[0044] Preferably, R2 is selected from hydrogen, halogens, or C atoms substituted with 1-3 halogens. 1-6 Alkyl, C 3-6 cycloalkyl; the C 3-6 The cycloalkyl group is optionally surrounded by 1-3 Rs. 5b Replaced;

[0045] More preferably, R2 is selected from 1-3 fluorine-substituted methyl or cyclopropyl groups; the cyclopropyl group may optionally be substituted with 1-3 halogens;

[0046] The preferred R2 is selected from monofluoromethyl or cyclopropyl.

[0047] In some embodiments, in the compounds of formula I, II, III, IV or V, or pharmaceutically acceptable salts thereof, R 5b Selected from halogens, C 1-6 Alkyl, C 3-6 Cycloalkyl, hydroxyl, cyano, amino, nitro; preferably R 5b Selected from halogen, methyl, ethyl, n-propyl, isopropyl, cyclopropyl, hydroxy, cyano, amino, nitro; more preferably R 5b Selected from fluorine, methyl, hydroxyl, cyano, amino, and nitro groups.

[0048] Secondly, this disclosure also provides a compound of the following formula or a pharmaceutically acceptable salt thereof.

[0049]

[0050] Thirdly, this disclosure also provides a compound of the following formula or a pharmaceutically acceptable salt thereof.

[0051]

[0052] Fourthly, this disclosure also provides an isotope-substituted product of a compound of formula I, II, III, IV, V, the second aspect, or the third aspect, or a pharmaceutically acceptable salt thereof, preferably, wherein the isotope substitution is deuterium substitution.

[0053] Fifthly, this disclosure also provides a pharmaceutical composition comprising a compound of formula I, II, III, IV, V, the second aspect, the third aspect, and the fourth aspect, or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient thereof.

[0054] In some embodiments, the unit dose of the pharmaceutical composition is 0.001 mg to 1000 mg.

[0055] In some embodiments, the pharmaceutical composition contains 0.01-99.99% of the aforementioned compound or a pharmaceutically acceptable salt thereof, based on the total weight of the composition. In some embodiments, the pharmaceutical composition contains 0.1-99.9% of the aforementioned compound or a pharmaceutically acceptable salt thereof. In some embodiments, the pharmaceutical composition contains 0.5%-99.5% of the compound or a pharmaceutically acceptable salt thereof. In some embodiments, the pharmaceutical composition contains 1%-99% of the compound or a pharmaceutically acceptable salt thereof. In some embodiments, the pharmaceutical composition contains 2%-98% of the compound or a pharmaceutically acceptable salt thereof. This disclosure also provides a method for treating or preventing HIV infection in humans by administering to a human a therapeutically effective amount of a compound or a pharmaceutically acceptable salt thereof as shown in Formula I, II, III, IV, V, the second aspect, the third aspect, the fourth aspect, or the pharmaceutical composition described in the fifth aspect.

[0056] This disclosure also provides a compound of formula I, II, III, IV, V, the second aspect, the third aspect, the fourth aspect, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, for the treatment or prevention of human HIV infection, wherein the compound or pharmaceutically acceptable salt thereof or pharmaceutical composition is used in combination with one or more other therapeutic agents.

[0057] In some embodiments, the additional therapeutic agent is selected from one or more of the following: HIV protease inhibitors, HIV non-nucleoside inhibitors of reverse transcriptase, HIV nucleoside inhibitors of reverse transcriptase, and HIV nucleotide inhibitors of reverse transcriptase.

[0058] In some embodiments, the additional therapeutic agent is selected from one or more of retegvir, lamivudine, abacavir, ritonavir, durugvir, darunavir, atazanavir, emtricitabine, tenofovir, ertigvir, rilpivirine, or lopinavir.

[0059] Although all the above structural formulas are drawn in some isomer form for simplicity, the present invention can include all isomers, such as tautomers, rotational isomers, geometric isomers, diastereomers, racemates, and enantiomers.

[0060] Tautomers are structural isomers of organic compounds that readily interconvert through a chemical reaction called tautomerization. This reaction often results in the migration of hydrogen atoms or protons, accompanied by the conversion of single bonds and adjacent double bonds. Some common tautomer pairs are keto-enol and lactam-lactamimide. An example of a lactam-lactamimide equilibrium is between A and B as shown below.

[0061]

[0062] All compounds in this invention can be designated as type A or type B. All tautomers are within the scope of this invention. The nomenclature of compounds does not exclude any tautomers.

[0063] "Optional" or "optional" means that the event or environment described below may but does not have to occur, and the description includes the possibility or absence of the event or environment. For example, "optionally halogenated or cyano-substituted C1-C6 alkyl" means that a halogen or cyano group may but does not have to be present, and the description includes cases where the alkyl group is substituted with a halogen or cyano group and cases where the alkyl group is not substituted with a halogen or cyano group.

[0064] In the chemical structure of the compound described in this invention, the bond... This indicates that the configuration is not specified; that is, if chiral isomers exist in the chemical structure, the bond... It can be or Or simultaneously include and Two configurations. In the chemical structure of the compounds described in this disclosure, the bonds... No configuration is specified, meaning it can be Z configuration, E configuration, or both configurations.

[0065] This disclosure covers the compounds described herein, their pharmaceutically acceptable salts, or any isotopically labeled derivatives of their isomers. Atoms capable of being isotopically labeled include, but are not limited to, hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, chlorine, and iodine. They can be labeled with isotopes. 2 H(D), 3 H, 11 C 13 C 14 C 15 N、 18 F, 31 P, 32 P, 35 S, 36 Cl and 125 I, etc. are used instead. Unless otherwise stated, when a position is specifically designated as deuterium (D), the position shall be understood as having a deuterium abundance of at least 3,000 times greater than the natural abundance of deuterium (which is 0.015%) (i.e., at least 45% deuterium incorporation).

[0066] Terminology Explanation:

[0067] "Pharmaceutical composition" means a mixture containing one or more of the compounds described herein or their physiologically pharmaceutically acceptable salts or prodrugs, along with other chemical components, such as physiologically pharmaceutically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to a living organism, thereby promoting the absorption of the active ingredient and its biological activity.

[0068] "Pharmaceutical excipients" include, but are not limited to, any adjuvants, carriers, excipients, flow aids, sweeteners, diluents, preservatives, dyes / colorants, flavorings, surfactants, wetting agents, dispersants, suspending agents, stabilizers, isotonic agents, solvents, or emulsifiers that have been approved by the U.S. Food and Drug Administration for use in humans or livestock.

[0069] "alkyl" refers to a saturated aliphatic hydrocarbon group, including straight-chain and branched groups with 1 to 20 carbon atoms. Alkyl groups containing 1 to 6 carbon atoms are also included. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, and their various branched isomers. Unless otherwise specified, alkyl groups may be substituted or unsubstituted. When substituted, the substituent can be substituted at any usable connection point, preferably one or more of the following groups, independently selected from halogens, deuterium, hydroxyl, oxo, nitro, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, C 2-6 alkenyloxy group, C 2-6 Acryloxy group, C 3-6 Cycloalkyl, 3- to 6-membered heterocycloalkyl, C 5-8 Cycloalkenyl, C 3-6 Cycloalkoxy, 3- to 6-membered heterocycloalkoxy, C 5-8 Cycloalkenyloxy, C 6-10 Aryl or 5- to 6-membered heteroaryl, wherein C 1-6 Alkyl, C 1-6 Alkoxy, C 2-6 alkenyloxy group, C 2-6 Acryloxy group, C 3-6 Cycloalkyl, 3- to 6-membered heterocycloalkyl, C 5-8 Cycloalkenyl, C 3-6 Cycloalkoxy, 3- to 6-membered heterocycloalkoxy, C 5-8 Cycloalkenyloxy, C 6-10 The aryl or 5- to 6-membered heteroaryl group may be optionally substituted by one or more groups selected from halogen, deuterium, hydroxyl, oxo, nitro, and cyano.

[0070] "Cycloalkyl" or "cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent, wherein the cycloalkyl ring contains 3 to 20 carbon atoms, preferably 3 to 8 carbon atoms. Non-limiting examples of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, etc.; polycyclic cycloalkyl groups include spirocyclic, fused-ring, and bridged-ring cycloalkyl groups.

[0071] The cycloalkyl ring may be fused to an aryl, heteroaryl, or heterocycloalkyl ring, wherein the ring connected to the parent structure is a cycloalkyl group, and non-limiting examples include indanyl, tetrahydronaphthyl, benzocycloheptyl, etc. Unless otherwise specified, the cycloalkyl group may be optionally substituted or unsubstituted. When substituted, the substituent is preferably one or more of the following groups, independently selected from deuterium, alkyl, alkenyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxyl, or carboxylic acid ester group.

[0072] "Heterocyclic group" or "heterocycle" refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon group containing 3 to 20 ring atoms, one or more of which are selected from nitrogen, oxygen, or S(O). m (where m is an integer from 0 to 2) heteroatoms, but excluding the ring portion of -OO-, -OS-, or -SS-, with the remaining ring atoms being carbon. Preferably, it contains 3 to 12 ring atoms, of which 1 to 4 are heteroatoms; more preferably, it contains 3 to 7 ring atoms. Non-limiting examples of monocyclic heterocyclic alkyl groups include pyrrolidinyl, imidazolyl, tetrahydrofuranyl, tetrahydrothiophenyl, dihydroimidazolyl, dihydrofuranyl, dihydropyrazolyl, dihydropyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, etc. Polycyclic heterocyclic alkyl groups include spirocyclic, fused-ring, and bridged-ring heterocyclic alkyl groups. Non-limiting examples of "heterocyclic alkyl" include:

[0073] etc.

[0074] Heterocyclic alkyl groups may be optionally substituted or unsubstituted. When substituted, the substituents are preferably one or more of the following groups, independently selected from: halogen, deuterium, hydroxyl, oxo, nitro, cyano, C. 1-6 Alkyl, C 1-6 Alkoxy, C 2-6 alkenyloxy group, C 2-6 Acryloxy group, C 3-6 Cycloalkyl, 3- to 6-membered heterocycloalkyl, C 5-8 Cycloalkenyl, C 3-6 Cycloalkoxy, 3- to 6-membered heterocycloalkoxy, C 5-8 Cycloalkenyloxy, C 6-10Aryl or 5- to 6-membered heteroaryl, wherein C 1-6 Alkyl, C 1-6 Alkoxy, C 2-6 alkenyloxy group, C 2-6 Acryloxy group, C 3-6 Cycloalkyl, 3- to 6-membered heterocycloalkyl, C 5-8 Cycloalkenyl, C 3-6 Cycloalkoxy, 3- to 6-membered heterocycloalkoxy, C 5-8 Cycloalkenyloxy, C 6-10 The aryl or 5- to 6-membered heteroaryl group may be optionally substituted by one or more groups selected from halogen, deuterium, hydroxyl, oxo, nitro, and cyano.

[0075] The heterocyclic group ring can be fused to an aromatic ring, a heteroaromatic ring, or a cyclic hydrocarbon, wherein the ring connected to the parent structure is a heterocyclic group, and non-limiting examples include:

[0076] wait.

[0077] "Being replaced by one or more A, B..." means that it can be replaced by a single or multiple substituents. When replaced by multiple substituents, it can be a plurality of identical substituents, or a combination of one or a plurality of different substituents. Attached Figure Description

[0078] Figure 1 Compounds 1 and 2 exhibit antiviral activity EC50 against WT (wild-type) strains.

[0079] Figure 2 The antiviral activity EC50 of compounds 1 and 2 against the G140S / Q148R strain.

[0080] Figure 3 The antiviral activity EC50 of compounds 1 and 2 against the Q148R / N155H strain.

[0081] Figure 4 The antiviral activity of compounds 1 and 2 against the E138K strain is EC50.

[0082] Figure 5 The antiviral activity EC50 of compounds 1 and 2 against the R263K strain.

[0083] Figure 6 The antiviral activity EC50 of compounds 1 and 2 against the Y143R / Q148H strain.

[0084] Figure 7 The antiviral activity EC50 of compounds 1 and 2 against the Q148H strain.

[0085] In the above figures, RAL represents raltegravir; CAB represents cabotegravir; the EC50 of the antiviral activity of CAB against each mutant virus strain is set to 1; **: significant difference from CAB with a p-value less than 0.05; N indicates the number of repeated tests. Detailed Implementation

[0086] The present disclosure is further described below with reference to embodiments, but these embodiments are not intended to limit the scope of the present disclosure.

[0087] Experimental methods in the embodiments of this disclosure that do not specify specific conditions are generally performed under conventional conditions or as recommended by the raw material or product manufacturer. Reagents whose specific source is not specified are commercially available conventional reagents.

[0088] The structure of the compound was determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). NMR shifts (δ) were expressed in 10⁻¹⁰ ohms. -6 The unit (ppm) is given. NMR measurements were performed using a Bruker AVANCE-400 NMR spectrometer. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD), with tetramethylsilane (TMS) as the internal standard. The spatial configurations of the optical isomers (isomers) of the compounds were further confirmed by measuring single-crystal parameters.

[0089] HPLC determinations were performed using a Waters ACQUITY ultra high performance LC, Shimadzu LC-20A systems, Shimadzu LC-2010HT series, or Agilent 1200LC high performance liquid chromatograph (ACQUITY UPLC BEH C18 1.7UM 2.1X50MM column, Ultimate XB-C18 3.0*150mm column, or Ultimate C18 2.1*30mm column).

[0090] MS measurements were performed using a Waters SQD2 mass spectrometer in positive / negative ion mode, with a mass scan range of 100–1200.

[0091] Chiral HPLC analysis was performed using the following columns: Chiralpak IC-3 100×4.6mm ID, 3µm; Chiralpak AD-3 150×4.6mm ID, 3µm; Chiralpak AD-3 50×4.6mm ID, 3µm; Chiralpak AS-3 150×4.6mm ID, 3µm; Chiralpak AS-3 100×4.6mm ID, 3µm; ChiralCel OD-3 150×4.6mm I.D., 3µm; Chiralcel OD-3 100×4.6mm ID, 3µm; ChiralCel OJ-H 150×4.6mm ID, 5µm; and Chiralcel OJ-3 150×4.6mm ID, 3µm.

[0092] Thin-layer chromatography silica gel plates are Yantai Huanghai HSGF254 or Qingdao GF254. The silica gel plates used in thin-layer chromatography (TLC) have a diameter of 0.15 mm to 0.2 mm, and the diameter of the silica gel plates used for thin-layer chromatography separation and purification products is 0.4 mm to 0.5 mm.

[0093] Rapid column purification systems use either the Combiflash Rf150 (TELEDYNE ISCO) or Isolara One (Biotage).

[0094] Normal column chromatography generally uses Yantai Huanghai silica gel of 100-200 mesh, 200-300 mesh or 300-400 mesh as the carrier, or Changzhou Santai pre-filled ultrapure normal phase silica gel column (40-63μm, 60, 12g, 25g, 40g, 80g or other specifications).

[0095] Reversed-phase column chromatography typically uses Changzhou Sante pre-packed ultrapure C18 silica gel columns (20-45μm). 40g, 80g, 120g, 220g or other sizes).

[0096] The high-pressure column purification system uses Waters AutoP, in conjunction with the Waters XBridge BEH C18 OBDPrep Column. 5μm, 19mm x 150mm or Atlantis T3 OBD Prep Column, 5μm, 19mm x 150mm.

[0097] Chiral preparation columns used were DAICL CHIRALPAK IC (250 mm * 30 mm, 10 μm) or Phenomenex-Amylose-1 (250 mm * 30 mm, 5 μm).

[0098] The known starting materials disclosed herein can be synthesized using or in accordance with methods known in the art, or can be purchased from companies such as Shanghai Titan Technology, ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, Accela ChemBio Inc, and Darui Chemicals.

[0099] Unless otherwise specified in the examples, the reactions can be carried out under an argon or nitrogen atmosphere.

[0100] Argon or nitrogen atmosphere refers to a reaction flask connected to an argon or nitrogen gas balloon with a volume of approximately 1L.

[0101] A hydrogen atmosphere refers to a reaction vessel connected to a hydrogen balloon with a volume of approximately 1L.

[0102] The pressurized hydrogenation reaction was performed using a Parr 3916EKX hydrogenator and a Qinglan QL-500 hydrogen generator or an HC2-SS hydrogenator.

[0103] The hydrogenation reaction is usually carried out under vacuum, filled with hydrogen gas, and repeated 3 times.

[0104] The microwave reaction was performed using a CEM Discover-S 908860 microwave reactor.

[0105] Unless otherwise specified in the examples, "solution" refers to an aqueous solution.

[0106] Unless otherwise specified in the examples, the reaction temperature is room temperature, which is 20℃~30℃.

[0107] In the examples, the reaction process was monitored using thin-layer chromatography (TLC). The volume ratio of the developing solvent used in the reaction, the eluent system used for column chromatography to purify the compound, and the developing solvent system for TLC were adjusted according to the different polarities of the compounds. Small amounts of basic or acidic reagents such as triethylamine and acetic acid could also be added for adjustment.

[0108] Example 1

[0109]

[0110] 1. Synthesis of compound 1b

[0111]

[0112] Under nitrogen protection, 1a (250.4 mg, 0.794 mmol, 1.0 eq) was added to a 10 mL reaction flask, followed by 2.5 mL of acetonitrile, then AcOH (227 μL) and MsOH (16 μL). After the addition was complete, the mixture was transferred to a 66 °C oil bath and stirred for approximately 20 h. LC-MS analysis was performed. Then, (S)-2-amino-2-cyclopropylethanol hydrochloride (153.3 mg, 1.114 mmol, 1.4 eq) was added. After approximately 5 min, potassium carbonate (158.3 mg, 1.145 mmol, 1.4 eq) and MeCN (2.1 mL) were added. The mixture was stirred for another 20 h at 66 °C. LC-MS analysis was performed again. Once the reaction was complete, the reaction was stopped, the reaction solution was cooled, concentrated to dryness under reduced pressure, and the residue was diluted with 1 N HCl and DCM. The mixture was separated, the aqueous phase was extracted with DCM, and the organic phases were combined and dried over anhydrous sodium sulfate. The sample was filtered and concentrated to obtain crude product 1b (240 mg). This was used directly in the next reaction. Ms(ESI): m / z 321.1 [M+1] + .

[0113] 2. Synthesis of compound 1d

[0114]

[0115] At room temperature, 1b (300 mg, 0.94 mmol, 1.0 eq) and 1c (158.4 mg, 1.1 mmol, 1.2 eq) were added to a 25 mL round-bottom flask, followed by 5 mL of DCM. While stirring, DIEA (302 mg, 2.34 mmol, 2.5 eq) and HATU (448.3 mg, 1.18 mmol, 1.3 eq) were added. The mixture was then stirred at room temperature under nitrogen balloon protection for 2-3 hours. LC-MS analysis was performed to confirm the reaction was complete. The reaction solution was then directly added to silica gel and stirred. After column chromatography, crude oil product 1d (786 mg) was obtained, with a yield of 188%, and was directly used in the next reaction. Ms(ESI): m / z 446.1 [M+1] + .

[0116] 3. Synthesis of Compound 1

[0117]

[0118] At room temperature, crude product 1d (739.2 mg, 1.0 eq), LiBr (368 mg, 4.24 mmol, 2.5 eq), and THF (10 mL) were added to a 50 mL round-bottom flask. After addition, the flask was transferred to an oil bath at 62-65 °C and stirred for about 24 h. LC-MS analysis was performed to confirm the reaction was complete. The reaction was stopped, the reaction solution was cooled, and the THF was removed by concentration. The residue was added to 3N HCl (3 mL) under ice bath conditions, and then diluted with DCM (3 mL). The mixture was separated, and the aqueous phase was extracted with DCM. The organic phases were combined, washed once with saturated sodium bicarbonate aqueous solution (1.5 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness under reduced pressure. The filtrate was purified by preparative HPLC and lyophilized to obtain compound 1 (270 mg). Ms(ESI): m / z 432.1 [M+1] + . 1 H-NMR (400MHz, CDCl3) δ10.32-10.29(t,1H),8.35(s,1H),7.39-7.33(q,1H),6.84-6.77(m,2H),5.44-5.41(m,1H),4.69-4.59(m,2H) ,4.44-4.40(q,1H),4.32-4.28(q,1H),4.02-3.98(q,1H),3.92-3.86(m,2H),1.07-1.03(m,1H),0.79-0.62(m,3H),0.37-0.30(m,1H).

[0119] Example 2

[0120]

[0121] 1. Synthesis of compound 2b

[0122]

[0123] At room temperature (20°C), 2e (503 mg, 2.6 mmol) (prepared according to literature WO2020167628) and HCl / dioxane (4 M, 2.6 mL, 10.4 mmol) were added to a 25 mL round-bottom flask and stirred at room temperature (20°C) for 16 hours. The reaction was monitored for completeness by LC-MS. The reaction solution was concentrated to give compound 2b (320 mg) in 95% yield, which was directly used in the next reaction.

[0124] 2. Synthesis of compound 2c

[0125]

[0126] Under nitrogen protection, 2a (399 mg, 1.266 mmol, 1.0 eq) was added to a 25 mL reaction flask, followed by 4 mL of acetonitrile, then AcOH (364 μL) and MsOH (28.8 μL). After the addition was complete, the mixture was transferred to a 66 °C oil bath and stirred for approximately 24 h. LC-MS analysis showed that less than 1% of the starting material remained. 2c (234 mg, 1.806 mmol, 1.4 eq) was then added, followed by potassium carbonate (284.6 mg, 2.059 mmol, 1.6 eq) after approximately 10 min. The mixture was stirred for another 21 h at 66 °C. LC-MS analysis confirmed the reaction was complete. The reaction was stopped, the reaction solution was cooled, and concentrated to dryness under reduced pressure. The residue was diluted with 1 N HCl and DCM, separated, and the aqueous phase was extracted with DCM. The combined organic phases were dried over anhydrous sodium sulfate. The mixture was filtered and concentrated to obtain crude product 2c (420 mg). This crude product was used directly in the next reaction step. Ms(ESI): m / z313.0[M+1] + .

[0127] 3. Synthesis of compound 2d

[0128]

[0129] At room temperature, 2c (450 mg, 1.44 mmol, 1.0 eq) and 2f (222.6 mg, 1.56 mmol, 1.1 eq) were added to a 50 mL round-bottom flask. 8 mL of DCM was added, and while stirring, DIEA (385 mg, 2.98 mmol, 2.1 eq) and HATU (641.5 mg, 1.68 mmol, 1.2 eq) were added. The mixture was then stirred at room temperature for 2-3 hours under nitrogen balloon protection. LC-MS analysis was performed to confirm the reaction was complete. The reaction solution was then directly mixed with silica gel and subjected to column chromatography to obtain crude oil product 2d (877 mg), with a yield of 139%, which was directly used in the next reaction. Ms(ESI): m / z 438.1 [M+1] + .

[0130] 4. Synthesis of Compound 2

[0131]

[0132] At room temperature, crude product 2d (754 mg), LiBr (386.3 mg, 4.448 mmol), and THF (10 mL) were added to a 50 mL round-bottom flask. After addition, the flask was transferred to an oil bath at 62-65 °C and stirred for about 24 h. LC-MS analysis was performed to confirm the reaction was complete. The reaction was stopped, the reaction solution was cooled, and the THF was removed by concentration. The residue was added to 3N HCl (3 mL) under ice bath conditions, and then diluted with DCM (3 mL). The mixture was separated, and the aqueous phase was extracted with DCM. The organic phases were combined and washed once with saturated sodium bicarbonate aqueous solution (1.5 mL). The mixture was dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated under reduced pressure to obtain 506 mg of crude product. After preparative HPLC purification and lyophilization, compound 2 (44 mg) was obtained. Ms(ESI): m / z 424.1 [M+1] + . 1 H-NMR (400MHz, CDCl3) δ10.26-10.20(m,1H),8.35(s,1H),7.39-7.31(m,1H),6.84-6.78(m,2H),5.33-5.30 (m,1H),5.10-5.07(q,0.5H),4.98-4.95(q,0.5H),4.65-4.41(m,6H),4.25-4.21(m,1H),3.97-3.91(m,1H).

[0133] Test Example 1:

[0134] The protective effect of the test compound against MT-4 cells infected with human immunodeficiency virus type 1 IIIB (HIV-1IIIB) was evaluated at different concentrations.

[0135] Materials and methods

[0136] The test compound was dissolved in DMSO at a concentration of 10 mM and stored at -20°C.

[0137] 1) Cytotoxicity assay of compounds: The test compounds were evaluated using a starting concentration of 100 nM and five consecutive semi-logarithmic dilutions. Each compound was added to the assay medium at twice the final desired test concentration. The compound was diluted to 1 mM in DMSO. Then, the compound was diluted to 200 nM (with 2 μL of the 1 mM stock solution added) in a drug dilution tube containing 998 μL of assay medium. 320 μL of the 200 nM solution was transferred to 680 μL of assay medium (semi-logarithmic dilution), for a total of five serial dilutions. 100 μL of each concentration was added to two replicates for cytotoxicity assessment, and one well was added for colorimetric assessment.

[0138] 2) Assay for the anti-HIV-1IIIB activity of the compounds: A starting concentration of 10 nM and five consecutive 5-fold dilutions of the compounds were used. Each compound was added to the assay medium at twice the final desired test concentration. 100 μL of each concentration was added to three replicate wells for potency assessment, with one well added for colorimetric evaluation. AZT, purchased from Sigma Aldrich (St. Louis, Missouri, USA), was evaluated as a control compound in the antiviral assay.

[0139] Cellular protection assay for HIV infection

[0140] Cell culture: MT-4 cells were passaged in T-75 flasks prior to use in the antiviral assay. Cells were passaged in a 1:2 ratio one day before the assay to ensure they were in the exponential growth phase at infection. Total and viable cell quantification was performed using a hemocytometer and trypan blue dye. The viable cell percentage used for the assay was greater than 95%. Cells were cultured at 5 x 10⁶ cells / mL. 4 The cells were resuspended in tissue culture medium and added in 50 μL volumes to a microtiter plate containing the drug.

[0141] Virus preparation: The virus used for this assay was the lymphotropic HIV-1IIIB strain. It was amplified in CEM-SS cells to prepare the viral stock solution. Pre-titrated viral aliquots were removed from the freezer (-80°C) and slowly thawed to room temperature in a biosafety cabinet. The virus was diluted in tissue culture medium; 50 μL of diluted virus produced 85% to 95% cell killing rate 6 days post-infection. Immediately, 50 μL of the diluted virus was added to a microtiter plate containing cells and the drug.

[0142] The efficacy and toxicity of the compounds were evaluated using XTT: After six days of incubation at 37°C in a 5% CO2 incubator, cell culture plates were added to the XTT (2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino)carbonyl]-2H-tetrazole hydroxide) dye for testing. The XTT solution was a 1 mg / mL RPMI 1640 stock solution prepared daily. A 0.15 mg / mL methyl phenazine sulfate (PMS) solution was prepared in PBS and stored at -20°C protected from light. An XTT / PMS stock solution was prepared before use by adding 40 μL of PMS per mL of XTT solution. 50 μL of XTT / PMS was added to each well of the plate, and the plate was re-incubated at 37°C for 4 hours. The plate was sealed with adhesive plate sealant and gently shaken or inverted several times to mix the soluble formazan product, and the absorbance was read at 450 / 650 nm.

[0143] Table 1. Antiviral efficacy and cytotoxicity data.

[0144] AZT (control compound) 2 >500 >250 cabotegravir 2.40 >100 >41.7 Compound 1 3.12 >100 >32.1 Compound 2 0.89 >100 >112

[0145] Test Example 2:

[0146] Inhibitor activity assays targeting pseudoviruses (G140S / Q148R, Q148R / N155H, E138K, Q148H, R263K, and Y143R / Q148H) containing common clinical mutation sites of HIV-1 integrase.

[0147] A method for detecting the anti-HIV-1 activity of compounds based on pseudoviruses.

[0148] Preparation of pseudoviruses: using lentiviral plasmid pNL4-3-Luc-R - E - Using (Hypergillus niger) as the backbone, an integrase point mutant plasmid was constructed. HEK293T cells (ATCC) were cultured at a density of 5 x 10⁻⁶ cells per well. 5 The cells were seeded in 6-well plates. After 24 hours, when the cell density reached 60%–80%, transfection was performed. pNL4-3-Luc-R was added to the transfection reagent at a specific ratio. - E - or pNL4-3-Luc-R - E - -IN-mutant plasmid and lentiviral packaging plasmid pMD2.G (Beijing Zhuangmeng). Cell transfection was performed according to the instructions of transfection reagent PEI25K (Maokang Biotechnology). After 48 hours, the supernatant was collected, centrifuged at 3000 rpm for 5 minutes, filtered through a 0.45 μm microporous membrane, aliquoted, and stored at -80℃ for later use. In HEK293T cells, the pseudovirus titer was detected based on luciferase activity.

[0149] Assay for the antiviral activity of the compound: The test compound was semi-logarithmically diluted with DMSO in a 96-well V-plate to prepare a 4-fold concentration solution. The virus was diluted with complete culture medium according to the titration results of different viruses. HEK293T cells were digested with trypsin to prepare 2 x 10⁻⁶ solutions. 5 / mL of cell suspension. In a 96-well plate, add 50µl of compound diluent, 50µl of virus diluent, and 100µl of cell suspension sequentially. The final compound concentration is 1000nM ± 0.1nM. Incubate the 96-well plate at 37°C in a 5% CO2 incubator. After 72 hours, carefully aspirate 100µL of culture supernatant from each well, add 25µL of Steady-Glo assay reagent (Promega, Cat#E2550) to each well in the dark, and perform assay using a Tecan (Type#Spark).

[0150] Table 2 shows the effects of compounds 1, 2, and cabotegravir on different mutant strains of EC. 50 EC 90 data

[0151]

[0152] For common clinical mutant strains of integrase (G140S / Q148R, Q148R / N155H, E138K, Q148H, R263K and Y143R / Q148H), compound 2 showed superior activity to cabotegravir, with significant differences.

[0153] Test Example 3: Inhibitor activity test against clinical HIV-1 strains.

[0154] Virus preparation: Clinical HIV-1 virus strains (Table 3) were obtained from the NIH AIDS Research and Reference Reagent Library in the United States. PBMCs were extracted from three fresh blood samples that were HIV and HBV seronegative, co-cultured, and activated with PHA-P (phytohemagglutinin). The cells and virus were incubated at 37°C / 5% CO2, and the supernatant of the cell culture after infection was collected and stored at -80°C. The viral titer was determined in the PBMCs from the three donors.

[0155] Detection of antiviral activity of compounds:

[0156] A RW / 92 / 009 C MW / 93 / 959 D UG / 92 / 001

[0157] Assay for the antiviral activity of the compounds: Frozen PBMCs were resuspended in fresh culture medium (1 x 10⁻⁶). 6 Add 50 μL of the compound (cells / mL) to each well of a round-bottom 96-well plate. Add 100 μL of culture medium containing twice the final concentration of the compound to each well, followed by 50 μL of virus solution (final infection dose: MOI = 0.002). After 7 days of culture, assess HIV-1 replication by detecting reverse transcriptase activity in the supernatant. Simultaneously prepare 96-well plates containing only the compound and cells for the determination of the compound's cytotoxicity.

[0158] In human primary PBMCs, the C2C of the compound 50 All were greater than 100 nM. For different subtypes of HIV-1 clinical isolates (RW / 92 / 009, MW / 93 / 959, UG / 92 / 001), compound 2 showed better activity than cabotegravir in human PBMCs.

[0159] Table 3: Antiviral activity data

[0160]

Claims

1. A compound or a pharmaceutically acceptable salt thereof, said compound having the following structure: 。 2. A pharmaceutical composition comprising the compound of claim 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

3. Use of the compound of claim 1 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of claim 2 in the preparation of a medicament for the treatment or prevention of human HIV infection.

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