Antiviral trisubstituted aromatic heterocyclic derivative as well as preparation method and application thereof

Abstract

The invention provides an antiviral trisubstituted heteroaromatic ring derivative and a preparation method and application thereof, the structure of the trisubstituted heteroaromatic ring derivative is shown as a formula (I), and the trisubstituted heteroaromatic ring derivative is a brand-new trisubstituted heteroaromatic ring derivative and has a remarkable antiviral effect. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of medical technology. Specifically, the present invention provides a tri-substituted aromatic heterocyclic derivative having a structure as shown in formula (I), a preparation method and use thereof. Background Art

[0002] The new coronavirus was first discovered in late 2019 and subsequently spread rapidly in many countries and regions. Its high mortality rate and complex clinical symptoms seriously threatened the lives and health of the people.

[0003] The most widely studied triazine ring is 1,3,5-triazine, and 1,3,5-triazine derivatives are reported to have a variety of biological activities and a wide range of industrial applications. Literature surveys show that 1,3,5-triazine derivatives have a multidimensional range of biological activities, such as anti-tuberculosis, anti-HIV and antiviral, antimalarial, anti-tumor, antifungal, antimicrobial and herbicidal activities.

[0004] Papain-like protease (PLpro) is an important means for coronaviruses to interfere with the host's UPS to evade host immunity. All coronaviruses can encode PLpro, and some coronaviruses can encode two types. SCoV-2, like SCoV and MERS-CoV, only encodes a single PLpro. It not only helps the virus evade host immunity, but is also crucial for viral transcription and replication. PLpro is similar to the structure of cysteine ​​DUBs. Like most DUBs, it consists of four different domains. There is a catalytic triad (Cys111 / His272 / Asp286) composed of Cys111 and acidic residues between the thumb and palm domains, which has a high degree of similarity with USP14 and HAUSP. In addition to the cysteine ​​residue (Cys111) at the catalytic position, the finger domain of PLpro also has four cysteine ​​residues that coordinate with Zn2+ ions. Among all the cysteine ​​residues in PLpro, Cys111 is the most easily oxidized and has a stronger reaction to electrophilic reagents.

[0005] PLpro is a virally expressed DUB that performs deubiquitination functions and has strong cross-recognition ability for UBLs. PLpro (35-kDa) is a domain of Nsp3 (215-kDa) that hydrolyzes the protein after the second Gly residue by recognizing the Leu-X-Gly-Gly peptide motif (X = Arg, Lys or Asn) of the substrate protein. Leu and X occupy the S4 and S3 subsites, respectively, and the two Gly residues occupy the S2 and S1 subsites, which are covered by the BL2 loop to form a narrow groove leading to the active site. The overall sequence of SCoV-2PLpro has 83% consistency with SCoV PLpro, mainly in the zinc finger domain. The catalytic active site is well conserved, and the changes in the "open" and "closed" conformations of the BL2 loop determine whether the substrate leads to the catalytic site.

[0006] There are drugs on the market for other COVID-19 targets (such as RdRp and 3CLpro), but there are currently no reports of SCoV2PLpro target inhibitors on the market. Therefore, the development of inhibitors against this target is of great significance for overcoming the virus's immune escape in host cells and developing safer and more effective inhibitors. Summary of the invention

[0007] In order to solve the above problems of the prior art, the object of the present invention is to provide a trisubstituted aromatic heterocyclic derivative, a pharmaceutically acceptable salt thereof, a tautomer thereof or a stereoisomer thereof, so as to screen out antiviral compounds having excellent performance in terms of effectiveness, safety and selectivity. Another object of the present invention is to provide a method for preparing the derivative, a pharmaceutically acceptable salt thereof, a tautomer thereof or a stereoisomer thereof.

[0008] The first aspect of the present invention provides a tri-substituted aromatic heterocyclic compound, and a pharmaceutically acceptable salt thereof, a tautomer thereof or a stereoisomer thereof, wherein the structure of the tri-substituted aromatic heterocyclic compound is shown in formula (I):

[0009]

[0010] in:

[0011] R1 is selected from the following group: substituted or unsubstituted C 1-10 Alkyl, substituted or unsubstituted C 3-10 Cycloalkyl, substituted or unsubstituted C 1-10 Alkyleneoxy-C 1-10 Alkyl, substituted or unsubstituted C 1-10 Alkylamino, substituted or unsubstituted C 1-6 Alkanoyl, substituted or unsubstituted C 6-14 Aryl, or substituted or unsubstituted C5-14 heteroaryl;

[0012] R2 is selected from the group consisting of hydrogen, substituted or unsubstituted C 1-10 Alkyl, substituted or unsubstituted C 1-10 Alkenyl, substituted or unsubstituted C 1-10 Alkynyl, substituted or unsubstituted C 3-10 heterocyclic group, -C(=O)R5, -C(=O)OR5, -C(=O)N(R5)2, -C 1-6 Alkylene-N(R5)2, -C 1-6 Alkylene-OR5; wherein R5 is selected from the group consisting of H, guanidinyl, substituted or unsubstituted C 1-6 Alkyl, saturated or partially unsaturated C 3-6 Cycloalkyl, 3-10 membered heterocyclic group, or C 6-10 Aryl; wherein the cyclic hydrocarbon group or heterocyclic group may be saturated or partially unsaturated, but not aromatic;

[0013] X1, X2, X3 are each independently selected from the group consisting of CH, N or C;

[0014] X, Y, and Z are each independently selected from the following groups: N, CR7; wherein R7 is selected from the following groups: H, halogen, C 1-4 Alkyl; and when X, Y or Z is CH, the attachment site may be located on the carbon atom (in which case H is replaced by the attachment site);

[0015] The E1 and E2 are selected from the following groups: chemical bond, -O-, -NH-, -S-, -CH2-, -CO-NH-, -NH-CO-, -NR6-, -NR6-CH2-, -CH2-NR6-, -CO-NH-CH2-; wherein the R6 is selected from the following group: H or C 1-4 alkyl;

[0016] R3 is selected from the following group: substituted or unsubstituted C 6-14 Aryl, substituted or unsubstituted C 5-14 Heteroaryl, substituted or unsubstituted C 4-9 Membered heterocyclic group, substituted or unsubstituted C 3-8 Carbocyclic, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 1-6 Alkoxy, substituted or unsubstituted amino, substituted or unsubstituted C 2-6 Alkylcarbonyl;

[0017] When E1 and E2 are each independently a chemical bond, R3 is a structure selected from the following group which is unsubstituted or substituted by 1 to 3 substituents:

[0018]

[0019] wherein the rings R3-A and R3-B are each independently a 4-10 membered heterocyclic ring, and the 4-10 membered heterocyclic ring may have 0, 1 or 2 heteroatoms selected from O or S; A is selected from the group consisting of hydrogen, C 1-6 Alkyl, C 1-6 Alkylene-OH, C 1-6 Alkylene NH2, -C(O)H, -C(O)-CH=CH2, -C(O)-C≡CH, -C(O)-C≡CC 1-3 Alkyl, -C(O)-CH2-X, C(O)-CHX2, -C(O)-CH=CH-C 1-3 Alkyl, -C(O)-CH=CH-R8, X is the following halogen: F, Cl, Br, R8 is -C(O)-OC 1-3 alkyl;

[0020] Unless otherwise specified, the substitution refers to substitution by one or more of the following substituents: halogen, amino, hydroxyl, C 1-6 Alkoxy, cyano, nitro, C 1-6 Alkyl, halogenated C 1-6 Alkyl, hydroxyl C 1-6 Alkyl, C 3-10 Cycloalkyl, 3-8 membered heterocyclic group containing 1-3 atoms selected from N, O and S, C 2- 6 acyl groups.

[0021] In another preferred embodiment, R1 is substituted or unsubstituted C 1-3 Alkyl, substituted or unsubstituted C 3-6 Cycloalkyl; preferably selected from the group consisting of substituted or unsubstituted methyl, ethyl, propyl; more preferably selected from substituted or unsubstituted methyl;

[0022] R2 is hydrogen, substituted or unsubstituted C 1-10 Alkyl, substituted or unsubstituted C 1-10 Alkenyl, substituted or unsubstituted C 1-10 Alkynyl, -C(=O)R5, -C(=O)OR5, -C(=O)N(R5)2, -C 1-6 Alkylene-N(R5)2, -C 1-6 Alkylene-OR5; wherein R5 is selected from the group consisting of H, guanidinyl, substituted or unsubstituted C 16 Alkyl, saturated or partially unsaturated C 3-6 Cyclic hydrocarbon, saturated or partially unsaturated C 3-10 Heterocyclic or C 6-10Aryl; wherein the cycloalkyl or heterocyclic group may be saturated or partially unsaturated, but not aromatic; preferably selected from the following group: hydrogen, -C(=O)R5, -C 1-6 Alkylene-N(R5)2, -C 1-6 Alkylene-OR5; wherein R5 is selected from the group consisting of H, guanidinyl, substituted or unsubstituted C 1-6 Alkyl; preferably selected from hydrogen, -C(=O)R5; wherein R5 is substituted or unsubstituted C 1-6 alkyl;

[0023] X1, X2, X3 are each independently selected from the following group: CH, N or C; preferably N;

[0024] X, Y, and Z are each independently selected from the following group: N or CR7; wherein R7 is selected from the following group: H, halogen, C 1-4 alkyl;

[0025] E1 and E2 are each independently selected from the following group: chemical bond, -O-, -NH-, -CH2-, -NH-CO-, -NR6-, -NR6-CH2-, -CH2-NR6-; wherein, R6 is selected from the following group: H or C 1-4 alkyl;

[0026] R3 is selected from the following group: substituted or unsubstituted C 6-14 Aryl, substituted or unsubstituted C 5-14 Heteroaryl, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted amino; preferably selected from the following group: substituted or unsubstituted C 6-14 Aryl, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted amino;

[0027] A is selected from the following group: hydrogen, C 1-6 Alkylene-OH, C 1-6 Alkylene-NH2, -C(O)H, -C(O)-CH=CH2, -C(O)-CH2-X, -C(O)-CHX2, -C(O)-CH=CH-C 1-3 Alkyl, -C(O)-CH=CH-R8, X is the following halogen: F, Cl, Br, R8 is -C(O)-OC 1-3 Alkyl; preferably selected from: hydrogen, -C(O)-CH=CH2, -C(O)-CH2-X, -C(O)-CHX2, -C(O)-CH=CH-C 1-3 Alkyl, X is the following halogen: F, Cl, Br, R8 is -C(O)-OC 1-3 alkyl;

[0028] Unless otherwise specified, the substitution refers to substitution by one or more of the following substituents: halogen, amino, hydroxyl, C 1-6 Alkoxy, cyano, nitro, C 1-6 Alkyl, halogenated C 1-6 Alkyl, hydroxyl C 1-6 Alkyl, C 3-10 Cycloalkyl, 3-8 membered heterocyclic group containing 1-3 atoms selected from N, O and S, C 2-6 Acyl.

[0029] In another preferred embodiment, the -E1-R3-E2 has a structure selected from the following group:

[0030]

[0031]

[0032] In another preferred embodiment, E1 and E2 are each independently selected from the following group: chemical bond, -NH-, -NH-CO-, -NR6-, -NR6-CH2-, -CH2-NR6-; wherein, R6 is selected from the following group: H or C 1-4 alkyl;

[0033] R3 is selected from the following group: substituted or unsubstituted C 6-10 Aryl, substituted or unsubstituted C 5-10 Heteroaryl, substituted or unsubstituted C 4-9 Membered heterocyclic group, substituted or unsubstituted C 3-8 Carbocyclic, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted amino, substituted or unsubstituted C 2-6 alkylcarbonyl; or

[0034] When E1 and E2 are each independently a chemical bond, R3 is unsubstituted or substituted by 1 to 3 substituents selected from the following structures:

[0035]

[0036] wherein rings R3-A and R3-B are each independently a 4-10 membered heterocyclic ring, and the 4-10 membered heterocyclic ring may have 0, 1 or 2 heteroatoms selected from O or S;

[0037] A is selected from the following group: hydrogen, -C(O)R9; wherein R9 is selected from the following group: substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 2-6 Alkenyl, substituted or unsubstituted C 2-6 Alkynyl.

[0038] In another preferred embodiment, R1 is selected from substituted or unsubstituted methyl; R2 is selected from hydrogen, -C(=O)OR5; wherein R5 is H, substituted or unsubstituted C 1-6 alkyl;

[0039] X1, X2, and X3 are each independently selected from N;

[0040] X, Y, and Z are each independently selected from the following group: N or C.

[0041] In another preferred embodiment, the -E1-R3-E2 has a structure selected from the following group:

[0042]

[0043] In another preferred embodiment, A is selected from the following groups:

[0044]

[0045] In a further preferred embodiment of the present invention, the compound is selected from the following group:

[0046]

[0047]

[0048]

[0049]

[0050]

[0051] In the second aspect of the present invention, there is provided a trisubstituted aromatic heterocyclic compound as described in the first aspect of the present invention, and a pharmaceutically acceptable salt, a tautomer or a stereoisomer thereof, which can be used to prepare a pharmaceutical composition for treating diseases caused by viral infection.

[0052] In another preferred embodiment, the disease is coronavirus, calicivirus or picornavirus infection.

[0053] In another preferred embodiment, SARS-CoV-2 virus, SARS-CoV virus, MERS-CoV virus.

[0054] The third aspect of the present invention provides a pharmaceutical composition comprising the trisubstituted aromatic heterocyclic compound described in the first aspect of the present invention, and a pharmaceutically acceptable salt, a tautomer or a stereoisomer thereof, and a pharmaceutically acceptable carrier and / or excipient.

[0055] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here. DETAILED DESCRIPTION

[0056] After long-term and in-depth research, the inventors have designed and prepared a series of compounds as shown in formula (I). The compounds have excellent virus inhibition effects and can therefore be used to prepare pharmaceutical compositions for treating viral infections. Based on the above findings, the inventors have completed the present invention.

[0057] the term

[0058] In the present invention, unless otherwise specified, the terms used have the general meanings well known to those skilled in the art.

[0059] As used herein, the term “C 1-10 The term "alkyl" refers to a straight or branched chain alkyl group having 1 to 10 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, or the like. 1-10 "Alkylene" refers to the C 1-10 The alkyl group loses another H atom to form a divalent group.

[0060] The term "C 3-10 "Cycloalkyl" refers to saturated and partially unsaturated cyclic hydrocarbon groups having 3 to 10 carbons. Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, and cyclooctyl. The term "C3-C6 cycloalkyl" has a similar meaning.

[0061] The term "C 3-8 "Carbocycle" refers to a saturated or partially unsaturated cyclic hydrocarbon group having 3 to 8 carbon atoms, the ring skeleton of which is all carbon atoms. The carbocycle may be saturated or partially unsaturated, but not aromatic.

[0062] The term "C 3-10 "Heterocyclic group" refers to a saturated and partially unsaturated cyclic group having 3 to 10 carbon atoms, one or more atoms in the skeleton of which are selected from the group consisting of N, O and S, wherein the ring N atom can be oxidized to NO, and the ring S atom can be oxidized to SO or SO2, and the remaining ring atoms are carbon. The heterocyclic group can be saturated or partially unsaturated, but does not have aromaticity.

[0063] The term "C 1-6 "Alkoxy" refers to a straight or branched alkoxy group having 1 to 6 carbon atoms, such as methoxy, ethoxy, propoxy, isopropoxy, or the like. 1-3 Expressions such as "alkoxy" and "oxy" have similar definitions.

[0064] The term "C 6-14 The term "aryl" refers to an aromatic group having 6 to 14 carbon atoms, including monocyclic or polycyclic aromatic groups, such as phenyl, naphthyl, or the like.

[0065] The term "C 5-14 The term "heteroaryl" refers to a heteroaryl group having 5 to 14 ring atoms (having 1 to 8 heteroatoms selected from N, O or S in the backbone), such as pyrrolyl, pyridyl, furanyl, or the like.

[0066] The term "substituted" refers to the replacement of one or more hydrogen atoms on a group by a substituent selected from the group consisting of halogen, amino, hydroxyl, C 1-6 Alkoxy, cyano, nitro, C 1-6 Alkyl, halogenated C1-6 Alkyl, hydroxyl C 1-6 Alkyl, C 3-10 Cycloalkyl, 3-8 membered heterocyclic group containing 1-3 atoms selected from N, O and S atoms.

[0067] Unless otherwise specified, in the present invention, all compounds appearing are intended to include all possible optical isomers, such as single chiral compounds, or mixtures of various chiral compounds (i.e., racemates). Among all compounds of the present invention, each chiral carbon atom can be optionally in R configuration or S configuration, or a mixture of R configuration and S configuration.

[0068] As used herein, the term "compound of the present invention" refers to a compound of Formula I. The term also includes various crystalline forms, pharmaceutically acceptable salts, hydrates or solvates of the compound of Formula I.

[0069] As used herein, the term "pharmaceutically acceptable salt" refers to a salt formed by a compound of the present invention and an acid or base that is suitable for use as a drug. Pharmaceutically acceptable salts include inorganic salts and organic salts. A preferred class of salts is a salt formed by a compound of the present invention and an acid. Suitable acids for forming salts include, but are not limited to, inorganic acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, phosphoric acid, formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, methanesulfonic acid, benzenesulfonic acid, benzenesulfonic acid, and acidic amino acids such as aspartic acid and glutamic acid.

[0070] Preparation of trisubstituted aromatic heterocyclic derivatives, namely compounds of formula (I)

[0071] Once the structures of the compounds of the present invention are known, the compounds of the present invention can be prepared from commercially available reagents using the synthetic methods and reaction schemes described herein, or using other reagents and conventional methods well known to those skilled in the art.

[0072] Several feasible schemes for the synthesis of the compound are briefly described below:

[0073] Solution 1:

[0074]

[0075] In the first step, the compound of general formula (I-1) undergoes an aromatic nucleophilic substitution reaction with R1-NH2 under alkaline conditions to obtain a compound of general formula (I-2);

[0076] In the second step, the compound of general formula (I-2) undergoes an aromatic nucleophilic substitution reaction with R2-Cl under alkaline conditions to obtain a compound of general formula (I-3);

[0077] The third step is to react the compound of the general formula (I-3) with an aromatic amine under alkaline conditions to undergo an aromatic nucleophilic substitution reaction or a catalytic coupling reaction to obtain a compound of the general formula (I-4);

[0078] The fourth step is to react the compound of the general formula (I-4) with R3-E under alkaline conditions to undergo an aromatic nucleophilic substitution reaction to obtain a compound of the general formula (I-5);

[0079] The fifth step is to obtain a compound of the general formula (I-5) by condensation or amidation reaction under alkaline conditions to obtain a compound of the general formula (I);

[0080] Solution 2

[0081]

[0082] In the first step, the compound of general formula (II-1) undergoes an aromatic nucleophilic substitution reaction with R1-NH2 under alkaline conditions to obtain a compound of general formula (II-2);

[0083] In the second step, the compound of general formula (II-2) undergoes an aromatic nucleophilic substitution reaction with R2-Cl under alkaline conditions to obtain a compound of general formula (II-3);

[0084] The third step is to react the compound of general formula (II-3) with R3-E under alkaline conditions to undergo an aromatic nucleophilic substitution reaction to obtain a compound of general formula (II-4);

[0085] The fourth step is to react the compound of the general formula (II-4) with an aromatic amine under alkaline conditions to undergo an aromatic nucleophilic substitution reaction or a catalytic coupling reaction to obtain a compound of the general formula (II-5);

[0086] The fifth step is to obtain a compound of the general formula (I) by condensation or amidation reaction of the compound of the general formula (II-5) under alkaline conditions;

[0087] In the above preparation method, the reagent providing alkaline conditions is selected from organic bases or inorganic bases, the organic bases are triethylamine, N,N-diisopropylethylamine, lithium diisopropylamide, sodium tert-butoxide, sodium methoxide and potassium tert-butoxide, and the inorganic bases are sodium hydride, potassium phosphate, sodium carbonate, potassium carbonate, potassium acetate, cesium carbonate, sodium hydroxide, potassium hydroxide, sodium bicarbonate and lithium hydroxide. One or more;

[0088] The reagent providing acidic conditions is one or more of hydrogen chloride, a 1,4-dioxane solution of hydrogen chloride, a methanol solution of hydrogen chloride, trifluoroacetic acid, formic acid, acetic acid, hydrochloric acid, sulfuric acid, methanesulfonic acid, nitric acid and phosphoric acid;

[0089] The metal catalyst is one or more of palladium / carbon, Raney nickel, tetrakistriphenylphosphine palladium, palladium dichloride, palladium acetate, [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (Pd(dppf)Cl2), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex, bistriphenylphosphine palladium dichloride (Pd(PPh3)Cl2) and tris(dibenzylideneacetone)dipalladium (Pd2(dba)3);

[0090] The ligand is one or more of 2-dicyclohexylphosphino-2,6'-dimethoxybiphenyl (SPhos), 4,5-bisdiphenylphosphino-9,9-dimethylxanthene (XantPhos), 2-dicyclohexylphosphino-2,4,6-triisopropylbiphenyl (XPhos), 2-dicyclohexylphosphino-2'-(N,N-dimethylamino)-biphenyl (DavePhos), 1,1'-bis(diphenylphosphino)ferrocene (Dppf) and 1,1'-binaphthyl-2,2'-bisdiphenylphosphine (BINAP), preferably 1,1'-binaphthyl-2,2'-bisdiphenylphosphine (BINAP);

[0091] The condensing agent includes, but is not limited to, one or more of dicyclohexylcarbodiimide (DCC), diisopropylcarbodiimide (DIC), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDCI), 2-(7-benzotriazole oxide)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (HATU), 2-(1H-benzotriazol L-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate (TBTU), 1-hydroxybenzotriazole (HOBt) and 1-propylphosphoric anhydride (T3P).

[0092] The above reaction is preferably carried out in a solvent, and the solvent used is one or more of N,N-dimethylformamide, N,N-dimethylacetamide, N-methylpyrrolidone, dimethyl sulfoxide, 1,4-dioxane, water, tetrahydrofuran, dichloromethane, 1,2-dichloroethane, methanol, ethanol, toluene, petroleum ether, ethyl acetate, n-hexane and acetone.

[0093] Pharmaceutical compositions and methods of administration

[0094] Since the compounds of the present invention have excellent viral inhibitory activity, the compounds of the present invention and their various crystal forms, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates, and pharmaceutical compositions containing the compounds of the present invention as the main active ingredient can be used to treat (stabilize, alleviate or cure) diseases caused by viral infections.

[0095] The pharmaceutical composition of the present invention comprises a safe and effective amount of the compound of the present invention and a pharmaceutically acceptable excipient or carrier. Wherein "safe and effective amount" means: the amount of the compound is sufficient to significantly improve the condition without causing serious side effects. Usually, the pharmaceutical composition contains 1-2000 mg of the compound of the present invention per dose, and more preferably, contains 10-200 mg of the compound of the present invention per dose. Preferably, the "one dose" is a capsule or tablet.

[0096] "Pharmaceutically acceptable carrier" refers to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with the compounds of the present invention and with each other without significantly reducing the efficacy of the compounds. Some examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as ), wetting agents (such as sodium lauryl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0097] There is no particular limitation on the administration of the compound or pharmaceutical composition of the present invention. Representative administration methods include (but are not limited to): oral, parenteral (intravenous, intramuscular or subcutaneous), and intratumoral.

[0098] Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, for example, starches, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, and acacia; (c) humectants, for example, glycerol; (d) disintegrants, for example, agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solubilizers, for example, paraffin; (f) absorption accelerators, for example, quaternary ammonium compounds; (g) wetting agents, for example, cetyl alcohol and glyceryl monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents.

[0099] Solid dosage forms such as tablets, pills, capsules, pills and granules can be prepared using coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifiers, and the release of the active compound or compounds in such compositions can be delayed in a certain part of the digestive tract. Examples of embedding components that can be used are polymeric substances and waxes. If necessary, the active compound can also be formed into microencapsulated form with one or more of the above-mentioned excipients.

[0100] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, the liquid dosage form may contain an inert diluent conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide and oils, in particular cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances.

[0101] Besides such inert diluents, the composition may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0102] Suspensions, in addition to the active compounds, may contain suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methanol and agar, or mixtures of these substances, and the like.

[0103] Compositions for parenteral injection may include physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

[0104] The compounds of the present invention can be administered alone or in combination with other pharmaceutically acceptable compounds (eg, chemical anticancer drugs).

[0105] When administered in combination, the pharmaceutical composition may also include one or more (2, 3, 4, or more) other pharmaceutically acceptable compounds (e.g., chemical anticancer drugs). One or more (2, 3, 4, or more) of the other pharmaceutically acceptable compounds (e.g., chemical anticancer drugs) may be used simultaneously, separately, or sequentially with the compounds of the present invention to treat cancer or related diseases.

[0106] When using the pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to a mammal (such as a human) in need of treatment, wherein the dosage during administration is a pharmaceutically effective dosage, and for a person weighing 60 kg, the daily dosage is usually 1 to 2000 mg, preferably 20 to 500 mg. Of course, the specific dosage should also take into account factors such as the route of administration and the health status of the patient, which are all within the skill of a skilled physician.

[0107] Example

[0108] The method of the present invention is described below by means of specific embodiments to make the technical solution of the present invention easier to understand and grasp, but the present invention is not limited thereto. 1 H NMR spectra were measured using a Bruker instrument (400 MHz), and chemical shifts were expressed in ppm using tetramethylsilane as an internal standard (0.00 ppm). 1 H NMR notation: s = singlet, d = doublet, t = triplet, q = quartet, m = multiplet, br = broad, dd = doublet of a doublet, dt = doublet of a triplet. Coupling constants, when given, are given in Hz.

[0109] The mass spectrum was obtained by LC / MS using ESI as the ionization method.

[0110] HPLC model: Agilent 1260, Thermo Fisher U3000; chromatographic column model: Waters xbrige C18 (4.6*150 mm, 3.5 μm); mobile phase: A: ACN, B: Water (0.1% H3PO4); flow rate: 1.0 mL / min; gradient: 5% A for 1 min, increase to 20% A within 4 min, increase to 80% A within 8 min, 80% A for 2 min, back to 5% A within 0.1 min; wavelength: 220 nm; column oven: 35°C.

[0111] The thin layer chromatography silica gel plate uses Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plate. The silica gel plate used in thin layer chromatography (TLC) adopts a specification of 0.2mm-0.3mm, and the specification used for thin layer chromatography separation and purification products is 0.4mm-0.5mm.

[0112] Column chromatography generally uses Yantai Huanghai Silica Gel 200-300 mesh silica gel as the carrier.

[0113] In the following examples, unless otherwise specified, all temperatures are in degrees Celsius. Unless otherwise specified, various starting materials and reagents are commercially available or synthesized according to known methods. Commercially available materials and reagents are used directly without further purification. Unless otherwise specified, commercial manufacturers include but are not limited to Sinopharm Group, J&K Technology Co., Ltd., TCI (Shanghai) Chemical Industry Development Co., Ltd., Shanghai Bid Pharmaceutical Technology Co., Ltd. and Shanghai Myrel Chemical Technology Co., Ltd.

[0114] CD3OD: deuterated methanol

[0115] CDCl3: deuterated chloroform

[0116] DMSO-d6: deuterated dimethyl sulfoxide

[0117] Pd2(dba)3: tris(dibenzylideneacetone)dipalladium

[0118] Pd(dppf)Cl2:[1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride

[0119] XantPhos: 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene

[0120] XPhos: 2-Dicyclohexylphosphino-2,4,6-triisopropylbiphenyl

[0121] HATU: 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethyluronium hexafluorophosphate

[0122] DMAP: 4-dimethylaminopyridine

[0123] DMF: N, N-dimethylformamide

[0124] THF: Tetrahydrofuran

[0125] DIPEA: N, N-diisopropylethylamine

[0126] Boc: tert-butyl carbonate

[0127] Cbz: benzyl formate

[0128] DHP: 3,4-dihydro-2H-pyran

[0129] DIBAL: diisobutylaluminum hydride

[0130] TEA: triethylamine

[0131] NaBH3CN: Sodium cyanoborohydride

[0132] NaH: Sodium hydride

[0133] DCM: dichloromethane

[0134] PE: Petroleum ether

[0135] EA: Ethyl acetate

[0136] TLC: Thin layer chromatography

[0137] HPLC: High Performance Liquid Chromatography

[0138] purity

[0139] &:and

[0140] Hydrogen atmosphere means that the reaction bottle is connected to a hydrogen balloon with a volume of about 1L.

[0141] Unless otherwise specified in the examples, the reaction temperature is room temperature, 20°C-30°C.

[0142] The reaction progress in the embodiment is monitored by thin layer chromatography (TLC), and the developing solvent used in the reaction, the system of eluent for column chromatography or the developing solvent system for thin layer chromatography used for purifying the compound includes: A: petroleum ether and ethyl acetate system; B: dichloromethane and methanol system; C: n-hexane: ethyl acetate; wherein the volume ratio of the solvent varies according to the polarity of the compound, and a small amount of acidic or alkaline reagent, such as acetic acid or triethylamine, can also be added for adjustment.

[0143] Pharmaceutical Chemical Synthesis

[0144] Intermediate 1

[0145] 4-Aminomethylpyrrolidin-2-one hydrochloride IN-1

[0146]

[0147] Step 1: 5-Nitro-1,2,3,4-tetrahydroisoquinoline W-13a

[0148] 5-Nitroisoquinoline (348 mg, 2 mmol) was dissolved in glacial acetic acid (5 mL), cooled to 0°C, sodium borohydride (378 mg, 10 mmol) was added in batches, and the temperature was raised to 25°C for 2 h after addition. The reaction was complete when detected by TLC. Ice water solution (20 mL) was added to the reaction solution, stirred for 10 minutes, extracted with 10 mL of ethyl acetate for 3 times, the organic phases were combined and concentrated, and the crude product was purified by silica gel column chromatography to obtain the title compound W-13a (250 mg, yield 70%) as a white solid.

[0149] LC-MS: m / z = 179.1 [M+H] +

[0150] Step 2 5-Nitro-1,2,3,4-tetrahydroisoquinoline-2-carboxylic acid-2-methylpropan-2-yl ester W-13b

[0151] Compound W-13a (200 mg, 1.12 mmol) and NaOH (46 mg, 1.14 mmol) were dissolved in dioxane / H2O (2 mL + 2 mL), and di-tert-butyl dicarbonate (490 mg, 2.24 mmol) was slowly added, and the mixture was reacted at 25°C for 2 hours. The reaction was complete after TLC detection. The reaction solution was directly concentrated, and the crude product was purified by silica gel column chromatography to obtain the title compound W-13b (305 mg, yield 98%) as a white solid.

[0152] LC-MS: m / z = 223.3 [M-55] +

[0153] 1 H NMR (400MHz, CDCl3) δ7.79(d,J=7.7Hz,1H),7.39–7.27(m,2H),4.63(s,2H),3.62(t,J=5.9Hz,2H),3.07(t,J=5.9Hz,2H),1.48(d,J=1.5Hz,9H).

[0154] Step 3 5-amino-1,2,3,4-tetrahydroisoquinoline-2-carboxylic acid-2-methylpropan-2-yl ester W-13c

[0155] Compound W-13b (200 mg, 0.72 mmol) was dissolved in methanol (5 mL), 10% Pd / C (20 mg) was added, and the air in the reaction bottle was replaced with a hydrogen balloon until the container was filled with hydrogen. The reaction was carried out at room temperature for 6 hours, and the reaction was complete after TLC detection. The crude product was purified by silica gel column chromatography to obtain the title compound W-13c (130 mg, yield 73%) as a white solid.

[0156] LC-MS: m / z = 192.3 [M-55] +

[0157] 1 H NMR (400MHz, CD3OD_SPE) δ6.94(t,J=7.7Hz,1H),6.61(dd,J=7.9,1.1Hz,1H),6.48(d, J=7.6Hz,1H),4.48(s,2H),3.66(t,J=6.1Hz,2H),2.57(t,J=6.0Hz,2H),1.48(s,9H).

[0158] Example 1

[0159] 3-(((6-(Isoquinolin-5-ylamino)-4-(methylamino)-1,3,5-triazacyclohexyl-2-yl(amino(methyl)aniline

[0160]

[0161] Step 1: 4,6-dichloro-N-methyl-1,3,5-triazine-2-amine 1a

[0162] Methylamine hydrochloride (1.83 g, 27.11 mmol) was added to a solution of cyanuric chloride 1a (5.0 g, 27.11 mmol) and DIPEA (7.01 g, 54.22 mmol) in tetrahydrofuran (100 mL) at 0°C. The mixture was reacted for 1 hour. TLC showed that the reaction was complete. The reaction solution was quenched with water (100 mL) and extracted with ethyl acetate (50 mL x 2). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by silica gel column chromatography (PE / EA=10 / 1) to obtain the title compound 1b (2.1 g, yield 43.27%) as a white solid.

[0163] LC-MS: m / z=179.0 [M+H] +

[0164] 1 H NMR (400MHz, DMSO) δ9.02–8.95 (m, 1H), 2.82 (d, J = 4.7Hz, 3H). 13C NMR (101MHz, DMSO) δ169.92,168.72,166.02,165.98,28.07,28.04.

[0165] Step 2 5-((6-chloro-4-(methylamino)-1,3,5-triazacyclohexyl-2-yl)amino)isoquinoline 1b

[0166] Compound 1a (2.1 g, 11.7 mmol) was dissolved in acetone / water (12 mL + 3 mL), and 5-aminoisoquinoline (1.7 g, 11.7 mmol) and potassium carbonate (2.4 g, 17.5 mmol) were added. After reacting at room temperature overnight, 20 mL of ice water was added and cooled and allowed to stand until the product was completely precipitated. The precipitated product was filtered and washed three times with cold water to obtain an off-white crude product (3.1 g, yield 93%).

[0167] LC-MS: m / z = 287.1 [M+H] +

[0168] Step 3: 3-(((6-(isoquinolin-5-ylamino)-4-(methylamino)-1,3,5-triazacyclohexyl-2-yl(amino(methyl)aniline

[0169] Compound 1b (565 mg, 1.98 mmol) was dissolved in DMF (5 mL), and ((3-(aminomethyl)phenyl)amino)methane acid-2-methylpropane-2-yl ester (460 mg, 2.08 mmol) and potassium carbonate (545 mg, 3.95 mmol) were added in sequence. The mixture was reacted at 90°C for 4 hours. The reaction was complete after TLC detection. 50 mL of water was added to the reaction solution, and the mixture was extracted three times with ethyl acetate. The organic phases were combined and concentrated, and the crude product was purified by silica gel column chromatography to obtain a white solid (620 mg, yield 66%). The purified solid was dissolved in 3 mL of dioxane, and 3 mL of 4M hydrochloric acid / dioxane solution was added. The reaction was stirred for 2 hours and concentrated to obtain the hydrochloride product.

[0170] LC-MS: m / z=373.2[M+H] +

[0171] Example 2

[0172] 5-((4-((Hexahydropyridin-4-ylmethyl)amino)-6-(methylamino)-1,3,5-triazacyclohexyl-2-yl)amino)isoquinoline 2

[0173]

[0174] Step 1: 4,6-dichloro-N-methyl-1,3,5-triazine-2-amine 1a

[0175] Methylamine hydrochloride (1.83 g, 27.11 mmol) was added to a solution of cyanuric chloride 1a (5.0 g, 27.11 mmol) and DIPEA (7.01 g, 54.22 mmol) in tetrahydrofuran (100 mL) at 0°C. The mixture was reacted for 1 hour. TLC showed that the reaction was complete. The reaction solution was quenched with water (100 mL) and extracted with ethyl acetate (50 mL x 2). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by silica gel column chromatography (PE / EA=10 / 1) to obtain the title compound 1a (4.2 g, yield 87%) as a white solid.

[0176] LC-MS: m / z=179.0 [M+H] +

[0177] 1 H NMR (400MHz, DMSO) δ9.02–8.95 (m, 1H), 2.82 (d, J = 4.7Hz, 3H). 13 CNMR(101MHz,DMSO)δ169.92,168.72,166.02,165.98,28.07,28.04.

[0178] Step 2 5-((6-chloro-4-(methylamino)-1,3,5-triazacyclohexyl-2-yl)amino)isoquinoline 1b

[0179] Compound 1a (2.1 g, 11.7 mmol) was dissolved in acetone / water (12 mL + 3 mL), and 5-aminoisoquinoline (1.7 g, 11.7 mmol) and potassium carbonate (2.4 g, 17.5 mmol) were added. After reacting at room temperature overnight, 20 mL of ice water was added and cooled and allowed to stand until the product was completely precipitated. The precipitated product was filtered and washed three times with cold water to obtain an off-white crude product (3.1 g, yield 93%).

[0180] LC-MS: m / z = 287.1 [M+H] +

[0181] Step 3: 5-((4-((hexahydropyridin-4-ylmethyl)amino)-6-(methylamino)-1,3,5-triazacyclohexyl-2-yl)amino)isoquinoline 2

[0182] Compound 1b (80 mg, 0.28 mmol) was dissolved in DMF (2 mL), and 4-(aminomethyl)piperidin-1-carboxylic acid-2-methylpropane-2-yl ester (63 mg, 0.29 mmol) and potassium carbonate (77 mg, 0.56 mmol) were added in sequence. The mixture was reacted at 90°C for 4 hours. The reaction was complete after TLC detection. 50 mL of water was added to the reaction solution, and the mixture was extracted three times with ethyl acetate. The organic phases were combined and concentrated, and the crude product was purified by silica gel column chromatography to obtain a white solid (100 mg, yield 78%). The purified solid was dissolved in 3 mL of dioxane, and 3 mL of 4M hydrochloric acid / dioxane solution was added. The reaction was stirred for 2 hours and concentrated to obtain the hydrochloride product.

[0183] LC-MS: m / z=365.2[M+H] +

[0184] Example 3

[0185] 2-Chloro-N-[3-(((6-(isoquinolin-5-ylamino)-4-(methylamino)-1,3,5-triazacyclohexan-2-yl]amino)methyl)phenyl)acetamide

[0186]

[0187] The reactant 1 (80 mg, 0.21 mmol) was dissolved in DMF (2 mL) and stirred for 30 minutes in an ice-water bath. Chloroacetyl chloride (24 mg, 0.21 mmol) was dissolved in 1 mL of DMF and placed in an ice bath. Then, the DMF solution of chloroacetyl chloride was slowly added dropwise to the reaction bottle within 30 minutes. Stirring was continued at 0°C for 1 hour. The reaction was complete after TLC detection. The crude product was purified by Pre-HPLC to obtain the final product 3 (41 mg, yield 43%)

[0188] LC-MS: m / z = 449.2 [M+H] +

[0189] Example 4

[0190] 2-Chloro-1-[4-(((6-(isoquinolin-5-ylamino)-4-(methylamino)-1,3,5-triazacyclohexan-2-yl]amino)methyl)piperidin-1-yl)ethan-1-one

[0191]

[0192] The reactant 2 (80 mg, 0.22 mmol) was dissolved in DMF (2 mL) and stirred for 30 minutes in an ice-water bath. Chloroacetyl chloride (25 mg, 0.22 mmol) was dissolved in 1 mL of DMF and placed in an ice bath. Then, the DMF solution of chloroacetyl chloride was slowly added dropwise to the reaction bottle within 30 minutes. Stirring was continued at 0°C for 1 hour. The reaction was complete after TLC detection. The crude product was purified by Pre-HPLC to obtain the final product 3 (45 mg, yield 47%)

[0193] LC-MS: m / z = 441.2 [M+H] +

[0194] Example 5

[0195] 2-Chloro-N-[3-(((4-(methylamino)-6-(quinolin-5-ylamino)-1,3,5-triazacyclohexan-2-yl)amino)methyl)phenyl]acetamide

[0196]

[0197] Step 1: 4,6-dichloro-N-methyl-1,3,5-triazine-2-amine 1a

[0198] Methylamine hydrochloride (1.83 g, 27.11 mmol) was added to a solution of cyanuric chloride 1a (5.0 g, 27.11 mmol) and DIPEA (7.01 g, 54.22 mmol) in tetrahydrofuran (100 mL) at 0°C. The mixture was reacted for 1 hour. TLC showed that the reaction was complete. The reaction solution was quenched with water (100 mL) and extracted with ethyl acetate (50 mL x 2). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by silica gel column chromatography (PE / EA=10 / 1) to obtain the title compound 1b (2.1 g, yield 43.27%) as a white solid.

[0199] LC-MS: m / z=179.0 [M+H] +

[0200] 1 H NMR (400MHz, DMSO) δ9.02–8.95 (m, 1H), 2.82 (d, J = 4.7Hz, 3H). 13 CNMR(101MHz,DMSO)δ169.92,168.72,166.02,165.98,28.07,28.04.

[0201] Step 2: 2-Methylpropan-2-yl {[3-({[4-chloro-6-(methylamino)-1,3,5-triazacyclohexan-2-yl]amino}methyl)phenyl]amino}methanoate 1c

[0202] Compound 1a (1.5 g, 8.4 mmol) was dissolved in acetone / water (8 mL + 2 mL), and ((3-(aminomethyl)phenyl)amino)methane-2-methylpropan-2-yl ester (2.0 g, 8.8 mmol) and potassium carbonate (1.7 g, 12.5 mmol) were added. After reacting at room temperature overnight, water was added and the aqueous phase was extracted with ethyl acetate. After extraction three times, the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by silica gel column chromatography to obtain the title compound 1c (2.5 g, yield 83%) as a white solid.

[0203] LC-MS: m / z = 309.1 [M-55] +

[0204] 1 H NMR (400MHz, DMSO) δ7.51–7.35(m,1H),7.29(t,J=10.1Hz,1H),7.18(t,J=7.9Hz,1H),6.95–6. 84(m,1H),4.40(dd,J=8.1,4.7Hz,2H),2.74(dd,J=9.3,4.0Hz,3H),1.46(s,9H),1.23(s,1H). 13 C NMR(101MHz,DMSO)δ166.18,153.25,140.39,139.98,128.96,121.81,121.09,117.78, 117.12,79.43,44.25,40.53,40.32,40.11,39.90,39.69,39.48,39.27,28.57,27.70.

[0205] Step 3: 2-Methylpropan-2-yl {[3-({[6-(methylamino)-4-(quinolin-5-ylamino)-1,3,5-triazacyclohexan-2-yl]amino}methyl)phenyl]amino}methanoate 5a

[0206] Compound 1c (200 mg, 0.55 mmol), 5-aminoquinoline (79 mg, 0.55 mmol), Pd2(dba)3 (100 mg, 0.11 mmol), Xanttphos (128 mg, 0.22 mmol), Cs2CO3 (536 mg, 1.65 mmol) were dissolved in 4 mL DMA and replaced with a balloon filled with nitrogen three times for five minutes each time. The reaction was heated at 100°C for 4 h under N2 protection. After the reaction was completed, 60 mL of water was added and the reaction solution was extracted with EA. After three extractions, the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by silica gel column chromatography to obtain the title compound 5a (138 mg, yield 53%) as a white solid.

[0207] LC-MS: m / z = 417.2 [M-55] +

[0208] Step 4: 2-chloro-N-[3-({[6-(methylamino)-4-(quinolin-5-ylamino)-1,3,5-triazacyclohexyl-2-yl]amino}methyl)phenyl]acetamide 5

[0209] Compound 5a (138 mg, 0.29 mmol) was dissolved in 2 mL of dioxane solution, and 2 mL of 4 M HCl / dioxane solution was added dropwise under magnetic stirring. The mixture was stirred for 2 h. After TLC showed that the reaction was complete, it was directly concentrated by rotary evaporator. The crude product was dissolved in 3 mL of DMF, and chloroacetyl chloride (32 mg, 0.29 mmol) was added dropwise under ice bath conditions within 30 minutes. The mixture was stirred for 2 h until the reaction was complete. The title compound 5 (45 mg, yield 35%) was obtained as a white solid after purification by Pre-HPLC.

[0210] LC-MS: m / z=449.2[M+H] +

[0211] 1 H NMR (500MHz, DMSO) δ9.03–8.95(m,1H),8.94–8.77(m,1H),8.09–8.03(m,1H),7.94(dd,J=17.2,8.7H z,2H),7.83–7.68(m,2H),7.60–7.01(m,4H),4.58(s,1H),4.18(d,J=10.5Hz,2H),3.08–2.82(m,3H).

[0212] Example 6

[0213] 2-Chloro-N-[3-({[6-(methylamino)-4-(quinolin-7-ylamino)-1,3,5-triazacyclohexan-2-yl]amino}methyl)phenyl]acetamide

[0214]

[0215] The experimental operation was the same as in Example 5. 6a was synthesized from intermediate 1c. The subsequent Boc removal and amidation reactions were the same. The product was purified by HPLC to obtain compound 6.

[0216] LC-MS: m / z=449.2[M+H] +

[0217] 1H NMR(500MHz,DMSO)δ10.33(d,J=11.2Hz,1H),9.07(q,J=9.4Hz,1H),8.97–8 .72(m,2H),8.26–8.05(m,2H),7.82–7.72(m,1H),7.60–7.42(m,2H),7.28(d ,J=8.3Hz,1H),7.13–7.04(m,1H),4.65(d,J=7.8Hz,2H),4.53(d,J=7.6Hz, 2H), 4.22 (t, J = 6.3Hz, 3H), 2.97 (s, 1H), 2.91 (s, 1H), 2.85 (d, J = 9.8Hz, 1H). 13 C NMR (126MHz, DMSO) δ165.09,159.47,159.20,158.92,158.65,138.99,130.10,129.82,129.21,1 25.11,124.84,123.94,123.07,120.13,119.59,118.38,117.79,115.46,113.12,62.29,44.02.

[0218] Example 7

[0219] 2-Chloro-N-[3-({[6-(methylamino)-4-(quinolin-8-ylamino)-1,3,5-triazacyclohexan-2-yl]amino}methyl)phenyl]acetamide

[0220]

[0221] The experimental operation was the same as in Example 5. 7a was synthesized from intermediate 1c. The subsequent Boc removal and amidation reactions were the same. The product was purified by HPLC to obtain compound 7.

[0222] LC-MS: m / z=449.2[M+H] +

[0223] 1 H NMR (500MHz, DMSO) δ10.38(s,1H),9.07–8.87(m,2H),8.44(dd,J=15.2,8.3Hz,1H),7.72–7.60(m,3H),7.62–7.44(m,2H),7.33( td,J=7.9,4.1Hz,1H),7.16–7.07(m,1H),4.61–4.54(m,2H),4.24(s,2H),4.02(s,2H),2.97–2.87(m,3H),1.23(d,J=5.5Hz,1H). 13C NMR (126MHz, DMSO) δ171.37,165.15,165.06,159.17,158.90,158.64,158.37,149.43,13 9.98,139.14,139.07,138.61,138.43,137.29,134.28,134.05,130.10,129.38,129.34, 129.14,128.33,128.29,127.45,127.33,123.80,123.15,122.82,122.77,122.59,122.25,120.42,119.05,118.65,118.45,117.99,117.71,115.72,62.31,44.40,44.04,43.99.

[0224] Example 8

[0225] 2-Chloro-N-[3-({[4-(isoquinolin-6-ylamino)-6-(methylamino)-1,3,5-triazacyclohexan-2-yl]amino}methyl)phenyl]acetamide 8

[0226]

[0227] The experimental operation was the same as in Example 5. 8a was synthesized from intermediate 1c. The subsequent Boc removal and amidation reactions were the same. The product was purified by HPLC to obtain compound 8.

[0228] LC-MS: m / z=449.2[M+H] +

[0229] 1 H NMR (400MHz, DMSO) δ10.38–10.32(m,1H),9.55(d,J=10.3Hz,1H),8.52–8.42(m,1H),8.35–8.27(m,1H),8.23–8.01(m,2H),7.94(s,1H),7 .49(d,J=8.5Hz,1H),7.30(td,J=8.0,3.8Hz,2H),7.12(d,J=7.8Hz,1H),4.63–4.54(m,2H),4.23(s,2H),2.93–2.80(m,3H),1.22(s,1H). 13C NMR (126MHz, DMSO) δ165.14,165.09,159.39,159.11,158.83,158.55,145.51,140.48,139.06,132.21,131.51,129.27, 124.94,123.63,123.44,122.64,120.06,118.29,117.92,117.72,115.39,113.05,112.35,62.31,44.03,43.99,43.64.

[0230] Example 9

[0231] 2-Chloro-N-[3-({[4-(isoquinolin-7-ylamino)-6-(methylamino)-1,3,5-triazacyclohexan-2-yl]amino}methyl)phenyl]acetamide 9

[0232]

[0233] The experimental operation was the same as in Example 5. 9a was synthesized from intermediate 1c. The subsequent Boc removal and amidation reactions were the same. The product was purified by HPLC to obtain compound 9.

[0234] LC-MS: m / z=449.2[M+H] +

[0235] 1 H NMR (400MHz, CD3OD_SPE) δ9.63–9.37(m,1H),9.12–8.76(m,1H),8.48(d,J=6.3Hz,1H),8.40–8.13(m,4H),7.83(s,1H ),7.36(t,J=7.1Hz,2H),7.24–7.17(m,1H),4.75(s,2H),4.18(d,J=7.6Hz,2H),3.12–3.01(m,3H),1.38–1.26(m,1H). 13 C NMR(126MHz,MeOD)δ166.15,146.78,140.20,138.64,138.26,135.11,131.62,130.60,128.94, 128.42,127.80,124.26,123.14,119.01,117.76,115.78,113.51,56.08,55.91,44.04,42.65.

[0236] Example 10

[0237] 2-Chloro-N-[3-({[4-(isoquinolin-8-ylamino)-6-(methylamino)-1,3,5-triazacyclohexan-2-yl]amino}methyl)phenyl]acetamide 10

[0238]

[0239] The experimental operation was the same as in Example 5. 10a was synthesized from intermediate 1c. The subsequent Boc removal and amidation reactions were the same. The product was purified by HPLC to obtain compound 10.

[0240] LC-MS: m / z=449.2[M+H] +

[0241] Embodiment 11

[0242] 2-Chloro-N-[3-({[6-(methylamino)-4-(quinolin-6-ylamino)-1,3,5-triazacyclohexyl-2-yl]amino}methyl)phenyl]acetamide 11

[0243]

[0244] The experimental operation was the same as in Example 5. 11a was synthesized from intermediate 1c. The subsequent Boc removal and amidation reactions were the same. The product was purified by HPLC to obtain compound 11.

[0245] LC-MS: m / z=449.2[M+H] +

[0246] 1 H NMR (400MHz, DMSO) δ10.40(d,J=5.6Hz,1H),9.01(td,J=6.0,2.8Hz,1H),8.73(d,J=18.0Hz,1H),8.16–8.05(m,2H),7.87–7.74(m,1H),7.6 5(d,J=15.6Hz,1H),7.50(d,J=8.1Hz,1H),7.31(t,J=7.8Hz,1H),7.12(d,J=7.5Hz,1H),4.71–4.55(m,2H),4.25(s,2H),3.04–2.87(m,3H). 13 C NMR (101MHz, DMSO) δ165.17,160.17,159.82,159.47,159.12,139.50,139.15,129.41,127. 78,125.14,122.78,122.51,120.95,118.59,118.03,115.12,112.20,44.01,28.01,27.98.

[0247] Example 12

[0248] 2-Chloro-N-[3-({[4-(isoquinolin-4-ylamino)-6-(methylamino)-1,3,5-triazacyclohexan-2-yl]amino}methyl)phenyl]acetamide 12

[0249]

[0250] The experimental operation was the same as in Example 5. 12a was synthesized from intermediate 1c. The subsequent Boc removal and amidation reactions were the same. The product was purified by HPLC to obtain compound 12.

[0251] LC-MS: m / z=449.2[M+H] +

[0252] Embodiment 13

[0253] 2-Chloro-N-[3-({[6-(methylamino)-4-(1,2,3,4-tetrahydroisoquinolin-5-ylamino)-1,3,5-triazacyclohexyl-2-yl]amino}methyl)phenyl]acetamide 13

[0254]

[0255] Step 1: 5-{[6-chloro-4-(methylamino)-1,3,5-triazacyclohexan-2-yl]amino}-1,2,3,4-tetrahydroisoquinoline-2-carboxylic acid-2-methylpropan-2-yl ester 13a

[0256] Compound 1a (120 mg, 0.67 mmol) was dissolved in acetonitrile (6 mL), and intermediate W-13c (166 mg, 0.67 mmol) and DIPEA (173 mg, 1.34 mmol) were added. The mixture was heated at 90°C for overnight reaction and then concentrated directly. The crude product was purified by silica gel column chromatography to obtain the title compound 13a (194 mg, yield 74%) as a white solid.

[0257] LC-MS: m / z=335.1[M-55] +

[0258] 1 H NMR (400MHz, DMSO) δ9.53–9.26(m,1H),7.99–7.73(m,1H),7.34–6.98(m,3H),4.51(s,2H),3.51(d,J=5.2Hz,2H),2.83–2.56(m,5H),1.43(s,9H). 13CNMR(101MHz,DMSO)δ168.48,166.43,166.30,164.96,154.40,136.34,136.29,131.53,1 26.44,126.19,125.26,124.50,79.43,60.23,28.56,28.54,27.72,24.91,21.21,14.54.

[0259] Step 2: 5-[(6-{[(3-aminophenyl)methyl]amino}-4-(methylamino)-1,3,5-triazacyclohexan-2-yl)amino]-1,2,3,4-tetrahydroisoquinoline-2-carboxylic acid-2-methylpropan-2-yl ester 13b

[0260] Compound 13a (150 mg, 0.38 mmol) was dissolved in acetonitrile (5 mL), and 3-(aminomethyl)aniline (70 mg, 0.57 mmol) and DIPEA (99 mg, 0.77 mmol) were added. The mixture was heated at 90°C for 6 h and then concentrated. Water was added and the mixture was extracted three times with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by silica gel column chromatography to obtain the title compound 13b (123 mg, yield 67%) as a white solid.

[0261] LC-MS: m / z = 421.3 [M-55] +

[0262] 1 H NMR (400MHz, CD3OD_SPE) δ7.50–7.25(m,1H),7.08(t,J=7.8Hz,1H),6.99(t,J=7.7Hz,1H),6.87(d,J=7.6H z,1H),6.70–6.47(m,3H),4.53–4.24(m,4H),3.51(t,J=6.0Hz,2H),2.79(s,3H),2.62(s,2H),1.46(s,9H). 13 C NMR(101MHz,CD3OD_SPE)δ165.96,164.79,155.15,147.32,140.63,136.65,134.01,128 .77,125.86,122.67,116.72,113.84,79.96,53.51,43.83,27.45,27.45,26.43,24.34.

[0263] Step 3: 5-({6-[({3-[(2-chloroacetyl)amino]phenyl}methyl)amino]-4-(methylamino)-1,3,5-triazacyclohexan-2-yl}amino)-1,2,3,4-tetrahydroisoquinoline-2-carboxylic acid-2-methylpropan-2-yl ester 13c

[0264] The reactant 13b (123 mg, 0.26 mmol) was dissolved in DMF (3 mL) and stirred for 30 minutes under ice-water bath. Chloroacetyl chloride (29 mg, 0.26 mmol) was dissolved in 1 mL of DMF and placed in ice-water bath. Then, the DMF solution of chloroacetyl chloride was slowly added dropwise to the reaction bottle within 30 minutes. Stirring was continued at 0°C for 1 hour. The reaction was complete after TLC detection. The crude product was purified by Pre-HPLC to obtain the final product 3 (50 mg, yield 35%)

[0265] LC-MS: m / z = 497.3 [M-55] +

[0266] Step 4: 2-chloro-N-[3-({[6-(methylamino)-4-(1,2,3,4-tetrahydroisoquinolin-5-ylamino)-1,3,5-triazacyclohexyl-2-yl]amino}methyl)phenyl]acetamide 13

[0267] The reactant 13c (50 mg, 0.09 mmol) was dissolved in DCM (2 mL), and 1 mL of TFA was slowly added to the reaction bottle, stirred at room temperature for 2 h, and the reaction was complete after TLC detection. The crude product was purified by Pre-HPLC to obtain the final product 3 (23 mg, yield 56%)

[0268] LC-MS: m / z=453.2[M+H] +

[0269] Embodiment 14

[0270] 2-Chloro-N-(4-{[4-(isoquinolin-5-ylamino)-6-(methylamino)-1,3,5-triazacyclohexan-2-yl]amino}butyl)acetamide 14

[0271]

[0272] The experimental operation was the same as that of Example 1 and Example 3. 14a was synthesized from the reaction intermediate 1b. The subsequent reaction with chloroacetyl chloride was the same as that of the above examples. The product was purified by HPLC to obtain compound 14.

[0273] LC-MS: m / z=415.2[M+H] +

[0274] 1H NMR(500MHz,MeOD)δ10.82–10.46(m,1H),9.58–9.32(m,1H),8.63–8.50(m,1H),8.00–7.69(m,2H),7.48–7.4 0(m,1H),4.04(d,J=9.1Hz,2H),3.66–3.41(m,3H),3.34–3.31(m,2H),3.13–2.95(m,4H),1.74–1.60(m,4H). 13 C NMR(126MHz,MeOD)δ167.98,166.51,166.17,145.45,132.54,127.85,127.57,126.15,126.04,120.02,119 .64,119.49,119.41,119.36,41.84,40.20,40.05,39.93,39.07,38.84,26.70,26.67,26.29,26.19,26.08.

[0275] Embodiment 15

[0276] 2-Chloro-N-[4-({4-[(2-chloroacetyl)amino]butyl}amino)-6-(isoquinolin-5-ylamino)-1,3,5-triazacyclohexan-2-yl]-N-methylacetamide 15

[0277]

[0278] The experimental operation was the same as that of Example 1 and Example 3. 14a was synthesized from the reaction intermediate 1b. In the subsequent reaction with chloroacetyl chloride, the equivalent of chloroacetyl chloride was increased to 2eq. The reaction was completed by TLC detection. The product was purified by HPLC to obtain compound 15.

[0279] LC-MS: m / z=491.1[M+H] +

[0280] 1 H NMR(500MHz,MeOD)δ11.03–10.80(m,1H),9.88–9.60(m,1H),8.69–8.53(m,2H),8.50–8.37(m,1H),8.13(td,J=8.0 ,2.8Hz,1H),4.47(d,J=2.2Hz,2H),4.04(d,J=8.1Hz,3H),3.69–3.44(m,4H),3.15–2.99(m,5H),1.77–1.60(m,6H). 13C NMR(126MHz,MeOD)δ167.97,167.63,146.28,134.55,133.44,131.54,130.36,1 28.91,127.75,120.74,42.32,41.83,40.23,39.06,38.77,26.71,26.28,26.08.

[0281] Example 16

[0282] 2-Chloro-N-(2-{[4-(isoquinolin-5-ylamino)-6-(methylamino)-1,3,5-triazacyclohexan-2-yl](methyl)amino}ethyl)-N-methylacetamide 16

[0283]

[0284] The experimental operation was the same as that of Example 1 and Example 3. 16a was synthesized from the reaction intermediate 1b. The subsequent reaction with chloroacetyl chloride was the same as that of the above examples. The product was purified by HPLC to obtain compound 16.

[0285] LC-MS: m / z=415.2[M+H] +

[0286] 1 H NMR(500MHz,MeOD)δ10.91–10.50(m,1H),9.55–9.40(m,1H),8.64–8.52(m,1H),8.10–7.76(m,2H),7.44(q,J=6.0Hz,1 H),4.30–4.12(m,2H),4.02–3.87(m,2H),3.79–3.65(m,2H),3.38–3.32(m,3H),3.25–3.13(m,3H),3.09–3.00(m,3H). 13 C NMR (126MHz, MeOD) δ167.71,165.56,146.13,145.33,132.56,132.37,128.05,126.14,125.93,120. 15,120.09,120.01,119.57,45.92,45.64,45.32,41.45,40.86,35.65,35.35,34.25,26.82,26.68.

[0287] Embodiment 17

[0288] 2-Chloro-N-[4-(7-chloro-5-methyl-6-oxoylidene-2,5-diazepin-2-yl)-6-(isoquinolin-5-ylamino)-1,3,5-triazacyclohexan-2-yl]-N-methylacetamide 17

[0289]

[0290] The experimental operation was the same as that of Example 1 and Example 3. 16a was synthesized from the reaction intermediate 1b. The equivalent of chloroacetyl chloride was increased to 2eq in the subsequent reaction with chloroacetyl chloride. The reaction was complete when detected by TLC. The product was purified by HPLC to obtain compound 17. LC-MS: m / z=491.1 [M+H] +

[0291] 1 H NMR (500MHz, DMSO) δ11.19–11.11(m,1H),11.06–10.81(m,1H),9.78–9.53(m,1H),8.95–8.69(m,2H),8.53–8.44(m,1H),8.15 (t,J=8.2Hz,1H),4.57(s,2H),4.48–4.22(m,3H),4.08–3.75(m,3H),3.35–3.15(m,4H),3.11–2.98(m,4H),2.96–2.87(m,3H). 13 C NMR (126MHz, DMSO) δ166.55,166.25,164.91,147.17,133.41,131.48,130.30,127.54,120.88,45.55,43.17,42.67,35.67,34.85,27.53.

[0292] Embodiment 18

[0293] 2-Chloro-1-{4-[6-(isoquinolin-5-ylamino)-4-(methylamino)-1,3,5-triazacyclohexan-2-yl]piperazin-1-yl}ethan-1-one 18

[0294]

[0295] The experimental operation was the same as that of Example 1 and Example 3. 18a was synthesized from the reaction intermediate 1b. The subsequent reaction with chloroacetyl chloride was the same as that of the above examples. The product was purified by HPLC to obtain compound 18.

[0296] LC-MS: m / z=413.2[M+H] +

[0297] 1H NMR (500MHz, DMSO) δ10.81–10.55(m,1H),9.67–9.26(m,1H),8.68(dd,J=14.6 ,7.3Hz,1H),8.27(dd,J=13.0,7.8Hz,1H),8.11–7.89(m,1H),7.84(td,J=8.0 ,1.8Hz,1H),7.41(d,J=7.8Hz,1H),4.49(t,J=5.9Hz,2H),4.11–4.04(m,2H), 3.99–3.83(m,2H),3.63(s,3H),3.05(d,J=4.8Hz,2H),2.94(d,J=4.7Hz,2H). 13 C NMR (126MHz, DMSO) δ165.45,158.57,146.66,145.90,137.73,133.06,128.15,127.42,120.89,119.29,85.34,61.78,42.48,40.43,28.05.

[0298] Embodiment 19

[0299] 2-Chloro-N-{1-[6-(isoquinolin-5-ylamino)-4-(methylamino)-1,3,5-triazacyclohexan-2-yl]piperidin-4-yl}acetamide 19

[0300]

[0301] The experimental operation was the same as that of Example 1 and Example 3. 19a was synthesized from the reaction intermediate 1b. The subsequent reaction with chloroacetyl chloride was the same as that of the above examples. The product was purified by HPLC to obtain compound 19.

[0302] LC-MS: m / z = 427.2 [M+H] +

[0303] 1 H NMR (500MHz, DMSO) δ10.77–10.58(m,1H),9.58–9.35(m,1H),8.67(dd,J=16.0,7.5Hz,1H),8.35(s,1H),8.26–8.15(m,1H),8.07–7.94(m,1H ),7.83(t,J=7.9Hz,1H),7.40(d,J=7.9Hz,1H),4.94–4.47(m,3H),4.07(d,J=3.7Hz,1H),3.03–2.92(m,4H),1.90(s,2H),1.46–1.16(m,4H).

[0304] Embodiment 20

[0305] 2-Chloro-N-{1-[6-(isoquinolin-5-ylamino)-4-(methylamino)-1,3,5-triazacyclohexyl-2-yl]azetidin-3-yl}acetamide 20

[0306]

[0307] The experimental operation was the same as that of Example 1 and Example 3. 20a was synthesized from the reaction intermediate 1b. The subsequent reaction with chloroacetyl chloride was the same as that of the above examples. The product was purified by HPLC to obtain compound 20.

[0308] LC-MS: m / z=399.1[M+H] +

[0309] 1 H NMR (500MHz, DMSO) δ10.73–10.47(m,1H),9.47–9.24(m,1H),9.17(d,J=6 .6Hz,1H),8.75–8.65(m,1H),8.39–8.25(m,1H),8.06–7.87(m,1H),7.81( td,J=8.0,1.8Hz,1H),7.40(d,J=7.8Hz,1H),4.65(s,2H),4.55(td,J=8.7 ,4.3Hz,2H),4.49–4.34(m,1H),4.14(d,J=1.6Hz,2H),3.02–2.88(m,3H). 13 C NMR (126MHz, DMSO) δ166.66,165.28,164.84,161.83,158.64,146.61,145.96,132.96,128.19, 127.04,121.00,119.56,56.97,42.95,40.37,40.21,40.04,39.87,39.71,39.54,39.37,27.99.

[0310] Embodiment 21

[0311] 2-Chloro-1-{6-[6-(isoquinolin-5-ylamino)-4-(methylamino)-1,3,5-triazacyclohexan-2-yl]-2,6-diazaspiro[3.3]hept-2-yl}ethan-1-one 21

[0312]

[0313] The experimental operation was the same as that of Example 1 and Example 3. 21a was synthesized from the reaction intermediate 1b. The subsequent reaction with chloroacetyl chloride was the same as that of the above examples. The product was purified by HPLC to obtain compound 21.

[0314] LC-MS: m / z=425.2[M+H] +

[0315] Embodiment 22

[0316] 2-Chloro-N-[4-({[6-(isoquinolin-5-ylamino)-4-(methylamino)-1,3,5-triazacyclohexan-2-yl]amino}methyl)phenyl]acetamide 22

[0317]

[0318] The experimental operation was the same as that of Example 1 and Example 3. 22a was synthesized from the reaction intermediate 1b. The subsequent reaction with chloroacetyl chloride was the same as that of the above examples. The product was purified by HPLC to obtain compound 22.

[0319] LC-MS: m / z=449.2[M+H] +

[0320] Embodiment 23

[0321] 2-Chloro-N-({4-[6-(isoquinolin-5-ylamino)-4-(methylamino)-1,3,5-triazacyclohexan-2-yl]-1,4-oxazepan-2-yl}methyl)acetamide 23

[0322]

[0323] The experimental operation was the same as that of Example 1 and Example 3. 23a was synthesized from the reaction intermediate 1b. The subsequent reaction with chloroacetyl chloride was the same as that of the above examples. The product was purified by HPLC to obtain compound 23.

[0324] LC-MS: m / z=443.1[M+H] +

[0325] Embodiment 24

[0326] (2E)-4-{[3-({[6-(isoquinolin-5-ylamino)-4-(methylamino)-1,3,5-triazacyclohexan-2-yl]amino}methyl)phenyl]amino}-4-oxobut-2-enoic acid methyl ester 24

[0327]

[0328] The experimental operation was the same as in Example 2. Product 1 was reacted with (2E)-4-chloro-4-oxyylidenebut-2-enoic acid methyl ester. The operation was the same as in the above example. The product was purified by HPLC to obtain compound 24.

[0329] LC-MS: m / z = 485.2 [M+H] +

[0330] Embodiment 25

[0331] N-[3-({[6-(isoquinolin-5-ylamino)-4-(methylamino)-1,3,5-triazacyclohexin-2-yl]amino}methyl)phenyl]prop-2-ynamide 25

[0332]

[0333] The experimental operation was the same as in Example 2. Product 1 was reacted with propioloyl chloride in the same manner as in the above example. The product was purified by HPLC to obtain compound 25.

[0334] LC-MS: m / z = 485.2 [M+H] +

[0335] Embodiment 26

[0336] N-[3-({[6-(isoquinolin-5-ylamino)-4-(methylamino)-1,3,5-triazacyclohexan-2-yl]amino}methyl)phenyl]prop-2-enamide 26

[0337]

[0338] The experimental operation was the same as in Example 2. Product 1 was reacted with propioyl chloride in the same manner as in the above example. The product was purified by HPLC to obtain compound 26.

[0339] LC-MS: m / z = 427.2 [M+H] +

[0340] Pharmacological activity test compound inhibitory effect on novel coronavirus PL protease

[0341] The volume of the entire enzymatic reaction system is 120 μL, the final concentration of PL protease is 50 nM, and the final concentration of substrate (RLRGG-AMC) is 20 μM. The buffer of the reaction system includes 50 mM TRIS pH 7.5, 0.1 mg / mL BSA. PL protease and the above compounds are added to a 96-well plate, two replicate wells for each concentration, incubated at room temperature for 20 minutes, the substrate is added and quickly placed in a microplate reader for reading. The excitation light and emission light are 360 ​​nm and 460 nm, respectively. The test time is 5 minutes, and the fluorescence value is read every 1 minute. The reaction rate is fitted according to the reading value, and compared with the control group (DMSO), the inhibition rate is calculated. The calculation formula is: inhibition rate = 1-(test group reaction rate / control group reaction rate).

[0342] The above conventional reagents were purchased from Sigma-Aldrich, the substrates were synthesized by GenScript Biotech, and the microplate reader model was Bio-tek Synergy H1.

[0343] Table 1 Inhibition rate and IC of the compounds of the present invention on the PL protease activity of the new coronavirus 50 value

[0344]

[0345]

[0346] Conclusion: The compounds of the present invention have a significant inhibitory effect on the enzymatic activity of SARS-CoV-2 PLpro protease.

[0347] All documents mentioned in the present invention are cited as references in this application, just as each document is cited as reference individually. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims

1. A trisubstituted aromatic heterocyclic compound, and a pharmaceutically acceptable salt thereof, a tautomer thereof or a stereoisomer thereof, characterized in that: The structure of the trisubstituted aromatic heterocyclic compound is shown in formula (I): in: R1 is selected from the following group: substituted or unsubstituted C 1-10 Alkyl, substituted or unsubstituted C 3-10 Cycloalkyl, substituted or unsubstituted C 1-10 Alkyleneoxy-C 1-10 Alkyl, substituted or unsubstituted C 1-10 Alkylamino, substituted or unsubstituted C 1-6 Alkanoyl, substituted or unsubstituted C 6-14 Aryl, or substituted or unsubstituted C 5-14 heteroaryl; R2 is selected from the group consisting of hydrogen, substituted or unsubstituted C 1-10 Alkyl, substituted or unsubstituted C 1-10 Alkenyl, substituted or unsubstituted C 1-10 Alkynyl, substituted or unsubstituted C 3-10 heterocyclic group, -C(=O)R5, -C(=O)OR5, -C(=O)N(R5)2, -C 1-6 Alkylene-N(R5)2, -C 1-6 Alkylene-OR5; wherein R5 is selected from the group consisting of H, guanidinyl, substituted or unsubstituted C 1-6 Alkyl, saturated or partially unsaturated C 3-6 Cycloalkyl, 3-10 membered heterocyclic group, or C 6-10 Aryl; wherein the cyclic hydrocarbon group or heterocyclic group may be saturated or partially unsaturated, but not aromatic; X1, X2, X3 are each independently selected from the group consisting of CH, N or C; X, Y, and Z are each independently selected from the following groups: N, CR7; wherein R7 is selected from the following groups: H, halogen, C 1-4 Alkyl; and when X, Y or Z is CH, the attachment site may be located on the carbon atom (in which case H is replaced by the attachment site); The E1 and E2 are selected from the following groups: chemical bond, -O-, -NH-, -S-, -CH2-, -CO-NH-, -NH-CO-, -NR6-, -NR6-CH2-, -CH2-NR6-, -CO-NH-CH2-; wherein the R6 is selected from the following group: H or C 1-4 alkyl; R3 is selected from the following group: substituted or unsubstituted C 6-14 Aryl, substituted or unsubstituted C 5-14 Heteroaryl, substituted or unsubstituted C 4-9 Membered heterocyclic group, substituted or unsubstituted C 3-8 Carbocyclic, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 1-6 Alkoxy, substituted or unsubstituted amino, substituted or unsubstituted C 2-6 Alkylcarbonyl; When E1 and E2 are each independently a chemical bond, R3 is a structure selected from the following group which is unsubstituted or substituted by 1 to 3 substituents: wherein rings R3-A and R3-B are each independently a 4-10 membered heterocyclic ring, and the 4-10 membered heterocyclic ring may optionally have 0, 1 or 2 heteroatoms selected from O or S; A is selected from the following group: hydrogen, C 1-6 Alkyl, C 1-6 Alkylene-OH, C 1-6 Alkylene -NH2, -C(O)H, -C(O)-CH=CH2, -C(O)-C≡CH, -C(O)-C≡CC 1-3 Alkyl, -C(O)-CH2-X, -C(O)-CH(X)2, -C(O)-CH=CH-C 1-3 Alkyl, -C(O)-CH=CH-R8, X is the following halogen: F, Cl, Br, R8 is -C(O)-OC 1-3 alkyl; Unless otherwise specified, the substitution refers to substitution by one or more of the following substituents: halogen, amino, hydroxyl, C 1-6 Alkoxy, cyano, nitro, C 1-6 Alkyl, halogenated C 1-6 Alkyl, hydroxyl C 1-6 Alkyl, C 3-10 Cycloalkyl, 3-8 membered heterocyclic group containing 1-3 atoms selected from N, O and S, C 2-6 Acyl.

2. The trisubstituted aromatic heterocyclic compound according to claim 1, and its pharmaceutically acceptable salt, its tautomer or its stereoisomer, characterized in that: R1 is substituted or unsubstituted C 1-3 Alkyl, substituted or unsubstituted C 3-6 Cycloalkyl; preferably selected from the group consisting of substituted or unsubstituted methyl, ethyl, propyl; more preferably selected from substituted or unsubstituted methyl; R2 is hydrogen, substituted or unsubstituted C 1-10 Alkyl, substituted or unsubstituted C 1-10 Alkenyl, substituted or unsubstituted C 1-10 Alkynyl, -C(=O)R5, -C(=O)OR5, -C(=O)N(R5)2, -C 1-6 Alkylene-N(R5)2, -C 1-6 Alkylene-OR5; Wherein, R5 is selected from the following group: H, guanidine, substituted or unsubstituted C 16 Alkyl, saturated or partially unsaturated C 3-6 Cyclic hydrocarbon, saturated or partially unsaturated C 3-10 Heterocyclic or C 6-10 Aryl; wherein the cycloalkyl or heterocyclic group may be saturated or partially unsaturated, but not aromatic; preferably selected from the following group: hydrogen, -C(=O)R5, -C 1-6 Alkylene-N(R5)2, -C 1-6 Alkylene-OR5; wherein R5 is selected from the group consisting of H, guanidinyl, substituted or unsubstituted C 1-6 Alkyl; preferably selected from hydrogen, -C(=O)R5; wherein R5 is substituted or unsubstituted C 1-6 alkyl; X1, X2, X3 are each independently selected from the following group: CH, N or C; preferably N; X, Y, and Z are each independently selected from the following group: N or CR7; wherein R7 is selected from the following group: H, halogen, C 1-4 alkyl; E1 and E2 are each independently selected from the following group: chemical bond, -O-, -NH-, -CH2-, -NH-CO-, -NR6-, -NR6-CH2-, -CH2-NR6-; wherein, R6 is selected from the following group: H or C 1-4 alkyl; R3 is selected from the following group: substituted or unsubstituted C 6-14 Aryl, substituted or unsubstituted C 5-14 Heteroaryl, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted amino; preferably selected from the following group: substituted or unsubstituted C 6-14 Aryl, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted amino; A is selected from the following group: hydrogen, C 1-6 Alkylene-OH, C 1-6 Alkylene-NH2, -C(O)H, -C(O)-CH=CH2, -C(O)-CH2-X, -C(O)-CHX2, -C(O)-CH=CH-C 1-3 Alkyl, -C(O)-CH=CH-R8, X is the following halogen: F, Cl, Br, R8 is -C(O)-OC 1-3 Alkyl; preferably selected from: hydrogen, -C(O)-CH=CH2, -C(O)-CH2-X, -C(O)-CHX2, -C(O)-CH=CH-C 1-3 Alkyl, X is the following halogen: F, Cl, Br, R8 is -C(O)-OC 1-3 alkyl; Unless otherwise specified, the substitution refers to substitution by one or more of the following substituents: halogen, amino, hydroxyl, C 1-6 Alkoxy, cyano, nitro, C 1-6 Alkyl, halogenated C 1-6 Alkyl, hydroxyl C 1-6 Alkyl, C 3-10 Cycloalkyl, 3-8 membered heterocyclic group containing 1-3 atoms selected from N, O and S, C 2-6 Acyl.

3. The trisubstituted aromatic heterocyclic compound according to claim 1, and its pharmaceutically acceptable salt, its tautomer or its stereoisomer, characterized in that: The -E1-R3-E2 has a structure selected from the following group:

4. The trisubstituted aromatic heterocyclic compound according to claim 1, and its pharmaceutically acceptable salt, its tautomer or its stereoisomer, characterized in that: In the compound: E1 and E2 are each independently selected from the following group: chemical bond, -NH-, -NH-CO-, -NR6-, -NR6-CH2-, -CH2-NR6-; wherein, R6 is selected from the following group: H or C 1-4 alkyl; R3 is selected from the following group: substituted or unsubstituted C 6-10 Aryl, substituted or unsubstituted C 5-10 Heteroaryl, substituted or unsubstituted C 4-9 Membered heterocyclic group, substituted or unsubstituted C 3-8 Carbocyclic, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted amino, substituted or unsubstituted C 2-6 alkylcarbonyl; or When E1 and E2 are each independently a chemical bond, R3 is unsubstituted or substituted by 1 to 3 substituents selected from the following structures: wherein rings R3-A and R3-B are each independently a 4-10 membered heterocyclic ring, and the 4-10 membered heterocyclic ring may have 0, 1 or 2 heteroatoms selected from O or S; A is selected from the following group: hydrogen, -C(O)R9; wherein R9 is selected from the following group: substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 2-6 Alkenyl, substituted or unsubstituted C 2-6 Alkynyl.

5. The trisubstituted aromatic heterocyclic compound according to claim 1, and its pharmaceutically acceptable salt, its tautomer or its stereoisomer, characterized in that: The compound -E1-R3-E2 has a structure selected from the group consisting of: or A is selected from the following groups:

6. The trisubstituted aromatic heterocyclic compound according to claim 1, and its pharmaceutically acceptable salt, its tautomer or its stereoisomer, characterized in that: The compound has a structure selected from the group consisting of:

7. The use of the trisubstituted aromatic heterocyclic compound, and the pharmaceutically acceptable salt thereof, or the tautomer or stereoisomer thereof according to any one of claims 1 to 6, characterized in that: Used for preparing a pharmaceutical composition for treating diseases caused by viral infection.

8. The use according to claim 7, characterized in that The disease is a coronavirus, calicivirus or picornavirus infection.

9. The use according to claim 7, characterized in that The disease is an infection caused by a virus selected from the group consisting of SARS-CoV-2 virus, SARS-CoV virus, and MERS-CoV virus.

10. A pharmaceutical composition, characterized in that The invention comprises the trisubstituted aromatic heterocyclic compound according to any one of claims 1 to 6, and a pharmaceutically acceptable salt thereof, a tautomer or a stereoisomer thereof, and a pharmaceutically acceptable carrier and / or excipient.

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