CN111606904A - Azaindoles and their applications - Google Patents

Abstract

The present invention provides azaindole compounds with the structure represented by formula (I) and applications thereof. The azaindole compounds can effectively inhibit SMAD3-phosphorylation, and have good antitumor activity in the rat LCC-induced tumor model, and compared with the compound SIS3, some of the azaindole compounds of the present invention Significantly improved water solubility. The azaindole compounds of the present invention are expected to be effective medicines for preventing and treating tumors.

Description

Azaindoles and their applications

technical field

The invention relates to the technical field of medicinal chemistry, in particular to an azaindole compound and an application thereof.

Background technique

Tumor growth and metastasis are the leading causes of death in cancer patients. During the invasive and metastatic steps of tumors, transforming growth factor beta (TGF-β) exerts tumor-promoting effects, and its inhibitory effect may lead to side effects due to the pleiotropic effects and complex functions of TGF-β, while downstream mediators of TGFβ signaling Because SMAD3 contains a DNA-binding domain, it can directly bind to promoters in target genes to regulate transcription, but SMAD2 and SMAD4 do not have these functions. In addition, SMAD3 has the ability to regulate the expression of key target proteins in tumor immunotherapy, for example, SMAD3 can upregulate the expression of programmed death receptor 1 (PD-1) on T cells, but SMAD2 cannot upregulate the expression of PD-1; the target Silencing of the protein SMAD3 can inhibit tumor cell growth and metastasis by enhancing the antitumor activity of NK cells and neutrophils in the tumor microenvironment. Therefore, developing effective and low-toxic drugs to selectively inhibit SMAD3 protein is a potential anti-tumor strategy.

There are still very few small-molecule compounds that selectively target and inhibit SMAD3, so it is of great significance to discover more types of SMAD3 protein-selective inhibitors.

SUMMARY OF THE INVENTION

Based on this, the present invention provides a new class of azaindole compounds, which can selectively inhibit SMAD3-phosphorylation, but not SMAD2, and can increase the proliferation of NK cells in the tumor microenvironment by increasing the These compounds have the potential to become small-molecule drugs for tumor immunotherapy.

The technical solutions are as follows:

Azaindoles having the structure represented by formula (I) or their pharmaceutically acceptable salts or their stereoisomers or their prodrug molecules:

in:

D is selected from: C, N; and when D is N, R is absent _;

A, B are independently selected from: N, CR ₃ ;

R ₁ is selected from: H, C ₁ -C ₆ alkyl, C ₆ -C ₁₀ aryl, C ₃ -C ₈ cycloalkyl, halogen, CN;

R ₂ is selected from: H, C ₁ -C ₆ alkyl;

Each R ₃ is independently selected from: H, C ₁ -C ₆ alkyl, halogen, CN;

R ₄ is selected from:

R ₅ , each R _5a and R _5b are each independently selected from: H, halogen, CN, C ₁ -C ₆ alkyl, C ₃ -C ₈ cycloalkyl;

Each R _is independently selected from:

R ₇ is selected from: H, C ₁ -C ₆ alkyl, C ₆ -C ₁₀ aryl substituted with one or more R ₁₃ , 6-10-membered heteroaryl substituted with one or more R ₁₃ ;

Each R ₈ is independently selected from: H, halogen, CN, C ₁ -C ₆ alkyl, C ₃ -C ₈ cycloalkyl, hydroxy, C ₁ -C ₆ alkoxy, C ₁ -C ₆ alkoxy substituted C ₁ -C ₆ alkoxy;

Each R ₉ is independently selected from: H, halogen, CN, C ₁ -C ₆ alkyl, C ₃ -C ₈ cycloalkyl, hydroxy, C ₁ -C ₆ alkoxy, C ₁ -C ₆ alkoxy substituted C ₁ -C ₆ alkoxy;

R ₁₀ is selected from: H, C ₁ -C ₆ alkyl;

R ₁₁ is selected from: hydroxyl, C ₁ -C ₆ alkoxy, -N(R ₁₄ )(R ₁₅ );

R ₁₂ is selected from: H, C ₁ -C ₆ alkyl, one or more R ₁₃ substituted C ₆ -C ₁₀ aryl, one or more R ₁₃ substituted 6-10-membered heteroaryl;

Each R ₁₃ is independently selected from: H, halogen, CN, C ₁ -C ₆ alkyl, C ₃ -C ₈ cycloalkyl, hydroxy, C ₁ -C ₆ alkoxy, C ₁ -C ₆ alkoxy substituted C ₁ -C ₆ alkoxy;

R ₁₄ and R ₁₅ are each independently selected from: H, C ₁ -C ₆ alkyl, amino substituted C ₁ -C ₆ alkyl, C ₁ -C ₆ alkylamino substituted C ₁ -C ₆ alkyl;

n is selected from: 1, 2, 3, 4;

m is selected from: 0, 1, 2, 3, 4;

p is selected from: an integer between 0-7;

q is selected from: 1, 2, 3, 4;

y is selected from: 1, 2, 3, 4, 5;

时，R₈不为C₁-C₆烷氧基。And, when D is C, A is N, B is CR ₃ , R ₁ is phenyl, and R ₂ is

, R ₈ is not C ₁ -C ₆ alkoxy.

In some of these embodiments, D is C, _A is N, and B is CR4.

In some of these embodiments, the azaindoles have the structure shown in formula (II):

_In some of these embodiments, R5 is H.

In some of these embodiments, n is selected from: 1, 2; m is selected from: 1 or 2.

In some of these embodiments, R ₇ is selected from: phenyl substituted with one or more R ₁₃ , six-membered nitrogen-containing heteroaryl substituted with one or more R ₁₃ .

In some of these embodiments, each R ₁₃ is independently selected from: H, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy.

In some of these embodiments, R ₇ is selected from:

In some embodiments, the azaindoles have the structure shown in formula (III):

In some of these embodiments, the azaindole compound has the structure shown in formula (IV):

In some of these embodiments, each R ₈ is independently selected from the group consisting of: H, hydroxy, C ₁ -C ₃ alkoxy, C ₁ -C ₃ alkoxy substituted C ₁ -C ₃ alkoxy.

In some of these embodiments, each R ₈ is independently selected from the group consisting of: hydroxy, methoxy, methoxy substituted ethoxy.

In some of these embodiments, R ₈ is both hydroxyl.

In some of these embodiments, the azaindoles have the structure shown in formula (V):

In some of these embodiments, R ₉ is selected from: H, halogen, hydroxyl;

R ₁₀ is selected from: H, C ₁ -C ₃ alkyl;

R ₁₁ is selected from: C ₁ -C ₆ alkoxy.

In some of these embodiments, the azaindole compounds have the structure shown in formula (VI):

In some of these embodiments, R ₁₂ is selected from: C ₆ -C ₁₀ heteroaryl substituted with one or more R ₁₃ ; each R ₁₃ is independently selected from: H, C ₁ -C ₆ alkyl.

In some of these embodiments, R ₁₂ is selected from:

In some of these embodiments, R _5a and R _5b are each independently selected from the group consisting of: H, halogen.

In some of these embodiments, R _5b is H and R _5a is F.

In some of these embodiments, R ₁ is selected from the group consisting of: H, C ₁ -C ₃ alkyl, C ₆ -C ₁₀ aryl, C ₃ -C ₆ cycloalkyl.

In some of these embodiments, R ₁ is selected from the group consisting of: H, phenyl, cyclopropyl.

In some of these embodiments, R ₂ is selected from: C ₁ -C ₃ alkyl.

In some of these embodiments, _R3 is H.

The present invention also provides the use of the above-mentioned azaindole compounds or their pharmaceutically acceptable salts or their stereoisomers or their prodrug molecules.

The specific technical solutions are as follows:

Use of the above-mentioned azaindole compounds or their pharmaceutically acceptable salts or their stereoisomers or their prodrug molecules in the preparation of SMAD3 inhibitors.

Use of the above-mentioned azaindole compounds or their pharmaceutically acceptable salts or their stereoisomers or their prodrug molecules in the preparation of medicaments for preventing and/or treating tumors.

In some of these embodiments, the tumor is: lung cancer, melanoma, breast cancer, liver cancer.

The present invention also provides a pharmaceutical composition for preventing and/or treating tumors.

The specific technical solutions are as follows:

A pharmaceutical composition for preventing and/or treating tumors, comprising an active ingredient and a pharmaceutically acceptable carrier, the active ingredient comprising the above-mentioned azaindole compound or a pharmaceutically acceptable salt thereof or a stereoisomer thereof Construct or its prodrug molecule.

The present study found a series of new azaindole compounds, which can effectively and selectively inhibit SMAD3-phosphorylation, but not SMAD2, and have good antitumor activity in a rat LCC-induced tumor model , and can enhance the anti-tumor effect by increasing the proliferation of NK cells in the tumor microenvironment.

In addition, the water solubility of pharmaceutical compounds is a major factor affecting the development of pharmaceutical preparations. Poor water solubility of the compounds may lead to low bioavailability, unstable metabolism (DrugDiscov.Today 2004, 9, 1020-1028), and the absorption of Fractional fluctuations (J.Med.Chem.2007,50,5858-5862), and problems such as difficulty in formulation development at later stages (Adv.DrugDeliv.Rev.1997,23,3-25), tend to make drugs The development time is greatly extended. The azaindole compounds or their salts of the present invention have good water solubility, and the solubility of HCl salts of some compounds in water exceeds 20 mg/mL, and even reaches 94 mg/mL or higher, which is beneficial to improve the bioavailability of the compounds degree, and is conducive to its later formulation development. The azaindole compounds of the present invention are expected to be effective medicines for preventing and treating tumors.

Description of drawings

Figure 1 is a graph showing the results of compound 16d selectively inhibiting TGF-β-dependent SMAD3 phosphorylation.

Figure 2 is a graph showing the results of compound 16d effectively inhibiting the growth of mouse syngeneic Lewis lung cancer (LLC).

Figure 3 is a graph showing the results of compound 16d inhibiting cancer progression by enhancing NK cell aggregation in LLC model mice.

Detailed ways

In the compounds of the present invention, when any variable (eg, R, etc.) occurs more than once in any component, its definition at each occurrence is independent of the definition at each other occurrence. Likewise, combinations of substituents and variables are permissible so long as such combinations stabilize the compound. A line drawn into a ring system from a substituent indicates that the indicated bond may be attached to any substitutable ring atom. If the ring system is polycyclic, it means that such bonds are only attached to any suitable carbon atoms adjacent to the ring. It will be appreciated that one of ordinary skill in the art can select substituents and substitution patterns for the compounds of the present invention to provide compounds that are chemically stable and readily synthesized from readily available starting materials by the skill of the art and the methods set forth below. If a substituent is itself substituted with more than one group, it is understood that these groups may be on the same carbon atom or on different carbon atoms, so long as the structure is stabilized.

The term "alkyl" as used herein is meant to include branched and straight chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms. For example, the definition of " _C1 - _C6 " in " _C1 - _C6 alkyl" includes groups having 1, 2, 3, 4, 5 or 6 carbon atoms in a straight or branched chain arrangement. For example, " _C1 - _C6 alkyl" specifically includes methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, pentyl, hexyl.

The term "cycloalkyl" refers to a monocyclic saturated aliphatic hydrocarbon group having the specified number of carbon atoms. For example, "cycloalkyl" includes cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, and the like.

The term "alkoxy" refers to groups having an -O-alkyl structure, such as -OCH ₃ , -OCH ₂ CH ₃ , -OCH ₂ CH ₂ CH ₃ , -O-CH ₂ CH(CH ₃ ) ₂ , - OCH ₂ CH ₂ CH ₂ CH ₃ , -O-CH(CH ₃ ) ₂ and the like.

The term "heteroaryl" refers to an aromatic ring containing one or more heteroatoms selected from O, N or S. Heteroaryl groups within the scope of the present invention include, but are not limited to: quinolinyl, pyrazolyl, pyrrolyl , thienyl, furanyl, pyridyl, pyrimidinyl, pyrazinyl, triazolyl, imidazolyl, oxazolyl, isoxazolyl, pyridazinyl; "heteroaryl" is also understood to include any nitrogen-containing N-oxide derivatives of heteroaryl groups.

The term "substituted" refers to the replacement of a hydrogen group in a particular structure with a group of the designated substituent.

As understood by those skilled in the art, "halo" ("halo") or "halo" as used herein means chlorine, fluorine, bromine and iodine.

Unless otherwise defined, alkyl, cycloalkyl, aryl, heteroaryl and heterocycloalkyl substituents can be unsubstituted or substituted. For example, a _C1 - _C6 alkyl group can be substituted with one, two, or three substituents selected from OH, halogen, alkoxy, dialkylamino, or heterocyclyl groups such as morpholinyl, piperidinyl, and the like.

The present invention includes the free forms of the compounds of formulae I-VI, as well as the pharmaceutically acceptable salts and stereoisomers thereof. Some of the specific exemplary compounds herein are protonated salts of amine compounds. The term "free form" refers to the amine compound in non-salt form. Included pharmaceutically acceptable salts include not only exemplary salts of the particular compounds described herein, but also typical pharmaceutically acceptable salts of all compounds of Formulas I-VI in free form. The free forms of specific salts of the compounds can be isolated using techniques known in the art. For example, the free form can be regenerated by treating the salt with a suitable dilute aqueous base, such as dilute aqueous NaOH, dilute aqueous potassium carbonate, dilute aqueous ammonia, and dilute aqueous sodium bicarbonate. The free forms differ somewhat from their respective salt forms in certain physical properties such as solubility in polar solvents, but for the purposes of the invention such acid and base salts are otherwise pharmaceutically equivalent to their respective free forms.

The pharmaceutically acceptable salts of the present invention can be synthesized from compounds of the present invention containing a basic or acidic moiety by conventional chemical methods. Generally, salts of basic compounds are prepared by ion exchange chromatography or by reacting the free base and a stoichiometric amount or excess of an inorganic or organic acid in the desired salt form in a suitable solvent or combination of solvents. Similarly, salts of acidic compounds are formed by reaction with a suitable inorganic or organic base.

Accordingly, pharmaceutically acceptable salts of the compounds of the present invention include conventional non-toxic salts of the compounds of the present invention formed by reacting a basic compound of the present invention with an inorganic or organic acid. For example, conventional non-toxic salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid, nitric acid, and the like, as well as those derived from organic acids such as acetic acid, propionic acid, succinic acid, glycolic acid, hard Fatty acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, pamoic acid, maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, p-aminobenzenesulfonic acid, 2-acetyl Salts prepared from oxymonobenzoic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethanedisulfonic acid, oxalic acid, isethionic acid, trifluoroacetic acid, etc.

If the compound of the present invention is acidic, appropriate "pharmaceutically acceptable salts" refer to salts prepared from pharmaceutically acceptable non-toxic bases including inorganic and organic bases. Salts derived from inorganic bases include aluminum, ammonium, calcium, copper, iron, ferrous, lithium, magnesium, manganese, manganous, potassium, sodium, zinc, and the like. Ammonium, calcium, magnesium, potassium and sodium salts are particularly preferred. Salts derived from pharmaceutically acceptable organic non-toxic bases including salts of primary, secondary and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins such as Amino acid, betaine, caffeine, choline, N,N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, aminoethanol, ethanolamine, ethyl Diamine, N-ethylmorpholine, N-ethylpiperidine, Glucosamine, Glucosamine, Histidine, Hydroxocobalamin, Isopropylamine, Lysine, Methylglucamine, Morpholine, Piperazine , Piperidine, quack, polyamine resin, procaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, etc.

Berg et al., "Pharmaceutical Salts," J. Pharm. Sci. '1977:66:1-19 describes the preparation of the pharmaceutically acceptable salts described above and other typical pharmaceutically acceptable salts in more detail.

Since deprotonated acidic moieties such as carboxyl groups in compounds can be anionic under physiological conditions, such charged charges can then be replaced by internally cationic protonated or alkylated basic moieties such as tetravalent The nitrogen atoms balance out, so it should be noted that the compounds of the present invention are potential inner salts or zwitterions.

In one embodiment, the present application provides a method for the treatment of hyperproliferative diseases or conditions such as tumors in humans or other mammals using compounds of formulas I-VI and pharmaceutically acceptable salts thereof.

In one embodiment, the compounds of the present application and pharmaceutically acceptable salts thereof can be used for the treatment or management of non-small cell lung cancer, small cell lung cancer, lymphoma, esophageal cancer, ovarian cancer, melanoma, cervical cancer, urinary tract cancer Skin cancer, pancreatic cancer, breast cancer, liver cancer, stomach cancer, bile duct cancer, leukemia, melanoma, colon cancer, rectal cancer, endometrial cancer or glioma.

Drug metabolites and prodrugs: the metabolites of the compounds involved in the application and their pharmaceutically acceptable salts, as well as prodrugs that can be converted into the structures of the compounds involved in the application and their pharmaceutically acceptable salts in vivo, also Included in the claims of this application.

Combinations: The compounds of formulae I-VI may be used in combination with other drugs known to treat or ameliorate similar conditions. When combined administration, the original drug administration mode & dosage remain unchanged, while the compound of formula I-VI is administered concurrently or subsequently. When a compound of formulae I-VI is administered concomitantly with one or more other drugs, it is preferred to use a pharmaceutical composition containing one or more known drugs and a compound of formulae I-VI. Drug combinations also include administration of a compound of formulae I-VI with one or more other known drugs at overlapping time periods. When the compounds of formulas I-VI are administered in combination with one or more other drugs, the doses of the compounds of formulas I-VI or known drugs may be lower than when they are administered alone.

Drugs or active ingredients that can be used in combination with the compounds of formulae I-VI include, but are not limited to:

Estrogen receptor modulators, androgen receptor modulators, retinal-like receptor modulators, cytotoxic/cytostatics, antiproliferative agents, protein transferase inhibitors, HMG-CoA reductase inhibitors, HIV protein kinase inhibitors agents, reverse transcriptase inhibitors, angiogenesis inhibitors, cell proliferation and survival signal inhibitors, drugs that interfere with cell cycle checkpoints and apoptosis inducers, cytotoxic drugs, tyrosine protein inhibitors, EGFR inhibitors, VEGFR inhibitors, serine/threonine protein inhibitors, Bcr-Abl inhibitors, c-Kit inhibitors, Met inhibitors, Raf inhibitors, MEK inhibitors, MMP inhibitors, topoisomerase inhibitors, histidine Acid deacetylase inhibitors, proteasome inhibitors, CDK inhibitors, Bcl-2 family protein inhibitors, MDM2 family protein inhibitors, IAP family protein inhibitors, STAT family protein inhibitors, PI3K inhibitors, AKT inhibitors , Integrin blocker, interferon-α, interleukin-12, COX-2 inhibitor, p53, p53 activator, VEGF antibody, EGF antibody, etc.

In one embodiment, the drugs or active ingredients that can be used in combination with the compounds of formulae I-VI include, but are not limited to: aldesleukin, alendronate, interferon, atranoin, allopurinol, allopurinol, Sodium purinol, Palonosetron hydrochloride, Hexamethylmelamine, Aminoglutide, Amifostine, Amrubicin, Amridine, Anastrozole, Dolassetron, aranesp, arglabin, Arsenic trioxide, Arsenic, 5-azacytidine, azathioprine, BCG or tice BCG, betadine, betamethasone acetate, betamethasone sodium phosphate preparation, bexarotene, bleomycin sulfate, bromuria Gan, bortezomib, busulfan, calcitonin, alezolizumab injection, capecitabine, carboplatin, casodex, cefesone, simoleukin, daunorubicin, chlorambucil, cisplatin, Cladribine, Cladribine, Clodronate, Cyclophosphamide, Cytarabine, Dacarbazine, Actinomycin D, Daunorubicin Liposome, Dexamethasone, Dexamethasone Phosphate, Valeric Acid Estradiol, Denisole-2, Dipomet, Deslorelin, Delazoxan, Diethylstilbestrol, Diflucan, Docetaxel, Deoxyfluridine, Doxorubicin, Dronabinol, Chin-166 -Chitosan complex, eligard, rasburicase, epirubicin hydrochloride, aprepitant, epirubicin, epoetin alfa, erythropoietin, eplatin, levamisole tablets, estradiol Preparation, 17-beta-estradiol, estramustine sodium phosphate, ethinyl estradiol, amifostine, hydroxyphosphate, fenbifur, etoposide, fadrozole, tamoxifen preparation, filgrastim , finasteride, feleastide, floxuridine, fluconazole, fludarabine, 5-fluorodeoxyuridine monophosphate, 5-fluorouracil, fluoxymesterone, flutamide, formin Stan, 1-β-D-arabinofuranosyl cytidine-5'-stearoyl phosphate, fomustine, fulvestrant, gamma globulin, gemcitabine, gemtuzumab, imabyl mesylate Matinib, Carzapine Wax Paper Capsules, Goserelin, Granisirone Hydrochloride, Histrelin, and Methionine, Hydrocortisone, Erythro-hydroxynonyladenine, Hydroxyurea, Titan Isobemumab, idarubicin, ifosfamide, interferon alpha, interferon-alpha2, interferon alpha-2A, interferon alpha-2B, interferon alpha-nl, interferon alpha-n3, interferon Interleukin β, Interferon γ-la, Interleukin-2, Intron A, Iressa, Irinotecan, Kaiteri, Lentinan sulfate, Letrozole, Leucovorin, Leuprolide, Leuprolide acetate, levothyroxine, levofolinate calcium salt, levothyroxine sodium, levothyroxine sodium preparations, lomustine, lonidamine, dronabinol, nitrogen mustard, methylcobalamin, Medroxyprogesterone acetate, megestrol acetate, melphalan, esterified estrogens, 6-mercaptopurine, mesna, methotrexate, methyl aminolevulinate, miltefosine, Meiman Amycin, mitomycin C, mitotane, mitosodium quinone, trilostane, doxorubicin citrate liposome, nedaplatin, pegylated filgrastim, oppreleukin, neupogen, nilutamide, tamoxifen, NSC-63 1570. Recombinant human interleukin-1-beta, octreotide, ondansetron hydrochloride, dehydrocortisone oral solution, oxaliplatin, paclitaxel, prednisone sodium phosphate preparation, pegmatase, perol Xin, pentostatin, streptolytic preparations, pilocarpine hydrochloride, pirorubicin, prukamycin, porfim sodium, prednimustine, steprednisolone, prednisone, Premarin, Procarba, Recombinant Human Erythropoietin, Raltitrexed, Ribbi, Etidronate, Rhenium-186, Rituxan, Rituxan-A, Romotide, Pilocarpine Hydrochloride Tablets, Octreotide, Shamo Krastim, Semustine, Sizoran, Sobuzoxan, Methylprednisolone, Paphos Acid, Stem Cell Therapy, Streptozocin, Strontium Chloride-89, Levothyroxine Sodium, Tamoxifen, tamsulosin, tasonamin, tastolactone, taxotere, tesetiazine, temozolomide, teniposide, testosterone propionate, methyltestosterone, thioguanine, thiotepa, thyrotropin, tiludosine acid, topotecan, toremifene, tosilimumab, trastuzumab, triosulfan, tretinoin, methotrexate tablets, trimethylmelamine, trimetrexate, acetic acid Triptorelin, Triptorelin pamoate, Eufodine, Uridine, Valrubicin, Vesrinone, Vinblastine, Vincristine, Vincamide, Vinorelbine, Velulizine, Dextropropimide, net statin, zofenin, paclitaxel protein stabilizer, acolbifene, interferon r-lb, affinitak, aminopterin, azoxifene, asoprisnil, atametan, atraxen Tan, BAY43-9006, Avastin, CCI-779, CDC-501, Celebrex, Cetuximab, Crinator, Cyproterone Acetate, Decitabine, DN-101, Doxorubicin -MTC, dSLIM, dutasteride, edotecarin, eflunomine, ixitecan, fenretinide, histamine dihydrochloride, histrelin hydrogel implant, holmium-166DOTMP, ibandronic acid, interferon gamma, intron-PEG, ixabepilone, keyhole limpet hemocyanin, L-651582, lanreotide, lasoxifene, libra, lonafamib, milepoxifene, minodex Ester, MS-209, liposomal MTP-PE, MX-6, nafarelin, naimrubicin, novarestat, nolatrexide, olimerson, onco-TCS, osidem, paclitaxel Polyglutamic acid ester, sodium bromide, PN-401, QS-21, quasiyang, R-1549, raloxifene, leopard enzyme, 13-cis-retinoic acid, saplatin, theocalcidol , T-138067, tarceva, docosahexaenoate paclitaxel, thymosin alfa, gazofurin, tipifarnib, tirapazamine, TLK-286, toremifene, trans MID-lo7R, valspro Dalta, vapretide, vatalanib, verteporfin, vinflunine, Z-100, and zolelinic acid or a combination thereof.

The following examples further describe the present invention, but the examples are not intended to limit the protection scope of the present invention.

Example 1: Synthesis of Compounds 5a-5j

Synthesis of Intermediate 4a:

NaH (192 mg, 4.8 mmol) was placed in the reaction flask, nitrogen was replaced, 7 mL of anhydrous DMF was added to dissolve, methyl phosphonoacetate diethyl ester was added under the condition of ice-water bath, stirred for 0.5 hours, compound 3a ( 636 mg, 3.973 mmol) in 7.5 mL DMF and stirred at room temperature for 8 hours. Sampling plate, the reaction is basically completed, add water to quench the reaction, stir for 2 hours, extract three times with ethyl acetate, wash with saturated brine, dry filter, concentrate and purify through silica gel column to obtain yellow solid compound n1 (520 mg, 60.5%); n1 (520 mg, 2.4 mmol) was dissolved in a mixed solution of THF/MeOH/H ₂ O (5 mL/8 mL/8 mL), NaOH (288 mg, 7.2 mmol) was added, and the mixture was stirred at room temperature for 4 hours. Sampling point plate, the reaction is basically completed. HCl (1M) was added to adjust pH=5.0, and intermediate 4a was obtained by filtration (413 mg, 85%).

Synthesis of Intermediate 4b:

To a solution of compound 3b (300 mg, 1.5 mmol) in pyridine (10 mL) was added malonic acid n2 (312 mg, 3 mmol) and piperidine (1 mL). The mixture was stirred at 110°C for 6 hours. The mixture was then cooled to room temperature and acidified with aqueous _NaHCO3 . Adjust pH=5 with HCl (1 M) and extract with EA. Washed with saturated brine, dried over anhydrous _MgSO4 , filtered, and concentrated to give crude product 4b as a yellow solid, which was used in the next step without further purification (188 mg, crude).

Synthesis of Intermediate 4c:

Compound 3c (360 mg, 1.52 mmol) and ethyl fluoroacetate (177 mg, 1.67 mmol) were placed in a reaction flask, replaced with nitrogen, dissolved in anhydrous DCM, and added dropwise TiCl ₄ (1M in CH ₂ Cl ₂ , 1.82 mL, 1.82 mmol), stirred at room temperature for 0.5 hours, then added TEA (0.42 mL, 3.02 mmol) and stirred at room temperature for 2 hours. Sampling plate, the reaction is basically completed, EA is extracted three times, the organic phases are combined, dried, filtered, concentrated, and purified by silica gel column chromatography to obtain compound n3 (220 mg, 47%).

¹ H NMR(400MHz, CDCl3)δ8.46-8.36(m,2H),7.61-7.51(m,3H),7.47-7.38(m,2H),7.22(dd,J=8.0,4.8Hz,1H) , 6.96 (d, J=40.1 Hz, 1H), 4.29 (q, J=7.1 Hz, 2H), 3.79 (s, 3H), 1.32 (t, J=7.1 Hz, 3H); ¹³ C NMR (100 MHz, CDCl3)δ161.9(d,J＝33.5Hz),148.7,144.1(d,J＝2.2Hz),143.9,143.8(d,J＝257.7Hz),130.8(d,J＝18.0Hz),130.7, 129.6, 129.5, 128.8, 118.4, 117.3 (d, J=3.5Hz), 111.5 (d, J=9.3Hz), 104.9 (d, J=2.5Hz), 61.4, 30.1, 14.3.

Compound n3 (220 mg, 0.71 mmol) and LiOH·H ₂ O (85 mg, 2.02 mmol) were placed in a reaction flask, THF and water (10 mL and 10 mL) were added. Stir at room temperature for 3 hours, sample and spot plate, the reaction is basically completed, acidified with HCl to pH 6, concentrated in vacuo, extracted three times with EA, combined organic phases, concentrated to give off-white solid Intermediate 4c (150 mg, 71%).

¹ H NMR (400MHz, DMSO-d6)δ8.36(d,J=4.4Hz,1H),8.21(d,J=7.8Hz,1H),7.66-7.47(m,5H),7.24(dd,J =7.9,4.6Hz,1H),6.76(d,J=40.5Hz,1H),3.66(s,3H); ^13C NMR(100MHz,DMSO-d6)δ162.8(d,J=33.9Hz), 148.6, 144.4 (d, J=255.7Hz), 144.3 (d, J=2.0Hz), 144.2, 131.2, 130.3 (d, J=18.0Hz), 130.1, 129.6, 129.3, 117.9, 117.8 (d, J= 3.3Hz), 110.7 (d, J=9.8Hz), 104.1 (d, J=2.4Hz), 30.2.

Synthesis of intermediate 4d:

A solution of compound 3c (300 mg, 1.27 mmol), tert-butyl 2-cyanoacetate (198 mg, 1.40 mmol), piperidine (36 mg, 0.42 mmol), acetic acid (174 mg, 2.90 mmol) and toluene (10 ml) was placed in the reaction bottle and stir for 15 hours. The sampling point plate reaction is basically completed. Extracted with EA (80 mL) and saturated aqueous NaHCO ₃ (30 mL), dried, filtered, concentrated, and purified by silica gel column chromatography to give compound n4 (410 mg, 90%) as a white solid.

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.82 (d, J=8.0 Hz, 1H), 8.47 (d, J=4.3 Hz, 1H), 8.08 (s, 1H), 7.63-7.55 (m, 3H) , 7.45–7.38(m, 2H), 7.34(dd, J=8.1, 4.7Hz, 1H), 3.83(s, 3H), 1.53(s, 9H). ¹³ C NMR(100MHz, CDCl ₃ )δ162.9,150.1 ,149.2,148.8,144.6,131.9,130.8,130.3,128.9,128.5,118.3,117.6,108.1,98.1,82.6,77.4,77.0,76.7,30.5,28.1.

Compound n4 (390 mg, 1.08 mmol) was placed in a reaction flask, 10 mL of CH ₂ Cl ₂ was added to dissolve, 3 mL of TFA was added, and the mixture was stirred at room temperature for 12 hours. The sampling point plate reaction is basically completed. Concentration gave 4d as a yellow solid (311 mg, 95%).

¹ HNMR (400MHz, DMSO-d ₆ )δ8.70(d,J=7.7Hz,1H),8.51(d,J=4.2Hz,1H),7.98(s,1H),7.71-7.57(m,5H) _The ^_ 128.4, 118.8, 118.5, 117.2, 107.4, 96.7, 30.8.

Synthesis of Intermediate 4e:

Using compound 3c (8.01 g, 33.90 mmol) as starting material, the intermediate 4e (8.03 g, 85%) was prepared according to the synthesis method of compound 4a.

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.45-8.31 (m, 2H), 7.68-7.55 (m, 5H), 7.48 (d, J=16.0 Hz, 1H), 7.32 (dd, J=7.9 , 4.8Hz, 1H), 6.40(d, J=16.0Hz, 1H), 3.69(s, 3H). ¹³ C NMR (100MHz, DMSO-d ₆ )δ168.8, 148.8, 145.7, 144.4, 137.7, 131.3, 130.2 ,129.6,129.4,129.3,118.3,117.9,114.6,108.0,30.1.HRMS(ESI)calcd for C ₁₇ H ₁₄ N ₂ O ₂ (M+H) ⁺ 279.1128,found279.1128.

Synthesis of Intermediate 4f:

Using compound 3d (940.4 mg, 4.0 mmol) as starting material, the intermediate 4f (747 mg, 55%) was prepared according to the synthesis method of compound 4a. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 7.94 (d, J=7.8 Hz, 1H), 7.65-7.55 (m, 4H), 7.53-7.43 (m, 3H), 7.31 (dt, J=24.9 ,7.5Hz,2H),6.33(d,J=16.0Hz,1H),3.62(s,3H).

Synthesis of Intermediate 4g:

Using compound 3e (300 mg, 1.27 mmol) as raw material, the intermediate 4g (200 mg, crude product) was prepared according to the synthesis method of compound 4b.

Synthesis of compound 5a

To a solution of compound 4a (126 mg, 0.623 mmol) and TEA (0.3 mL) in DMF (3 mL) was added HATU (237 mg, 0.52 mmol), stirred at 0°C for 0.5 h, and added 6,7-dimethoxy-1,2 , 3,4-Tetrahydroisoquinoline hydrochloride (120 mg, 0.623 mmol). Stir at room temperature for 5 hours, sample and spot plate, the reaction is basically completed, add ice water for dilution, suction filtration, dissolve in ethyl acetate, dry over anhydrous sodium sulfate, filter and concentrate, and purify with silica gel column to obtain compound 5a as a white solid (117 mg, 50%) .

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.39 (dd, J=4.7, 1.4 Hz, 1H), 8.18 (d, J=7.4 Hz, 1H), 7.88 (d, J=15.3 Hz, 1H), 7.47 (s,1H),7.19(dd,J=7.9,4.7Hz,1H),6.89(d,J=15.4Hz,1H),6.66(d,J=7.8Hz,2H),4.79(s,2H) , 3.89(s, 5H), 3.86(s, 6H), 2.87(t, J=28.1Hz, 2H). ¹³ CNMR(100MHz, CDCl3)δ166.7,148.8,147.9,143.9,135.8,132.3,128.5,127.3, ^found _{_ _ _ _} 378.1804.

Synthesis of compound 5b

To a solution of compound 4b (200 mg, 0.826 mmol) and 6,7-dimethoxy-1,2,3,4-tetrahydroisoquinoline hydrochloride (140 mg, 0.60 mmol) in DMF (15 mL) was added DIPEA (1 mL) and HATU (380 mg, 1 mmol). Stir at room temperature for 16 hours. Sampling point plate, the reaction is basically completed, diluted with water, EA extraction. The combined organic phases were washed with saturated brine, dried over anhydrous MgSO ₄ , filtered, concentrated, and purified by silica gel column chromatography to obtain compound 5b (90 mg, 26%) as a yellow solid product.

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.36 (dd, J=4.7, 1.4 Hz, 1H), 8.27 (d, J=15.4 Hz, 1H), 8.14 (d, J=7.2 Hz, 1H), 7.22 -7.14(m, 1H), 6.88(d, J=15.4Hz, 1H), 6.75-6.63(m, 2H), 4.82(s, 2H), 4.05-3.77(m, 11H), 3.00-2.80(m , 2H), 2.01–1.87 (m, 1H), 1.35–1.22 (m, 2H), 0.97–0.85 (m, 2H). ¹³ C NMR (100MHz, CDCl ₃ )δ167.0, 148.3, 144.6, 143.1, 135.7, _{127.8,127.3,126.1,125.9,124.4,118.4,116.8,112.9,112.7,111.5,111.2,109.5,109.2,56.0,47.2,44.5,43.8,40.1,28.3,6.7,6.3.HRMS} H ₂₇ N ₃ O ₃ (M+H) ⁺ 418.2125, found418.2128.

Synthesis of compound 5c

To a solution of compound 4c (130 mg, 0.44 mmol) and 6,7-dimethoxy-1,2,3,4-tetrahydroisoquinoline hydrochloride (111 mg, 0.48 mmol) in DMF (8 mL) was added DIPEA (0.23 mL, 1.32 mmol) and HATU (200 mg, 0.53 mmol). Stir at room temperature for 1 hour. Sampling point plate, the reaction is basically completed, diluted with water, EA extraction. The combined organic phases were washed with saturated brine, dried over anhydrous MgSO ₄ , filtered, concentrated, and purified by silica gel column chromatography to obtain compound 5c (165 mg, 79%) as a white solid product.

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.43 (d, J=4.7 Hz, 1H), 8.37 (d, J=7.2 Hz, 1H), 7.58-7.47 (m, 3H), 7.46-7.39 (m, 2H), 7.23 (dd, J=7.9, 4.8Hz, 1H), 6.78–6.55 (m, 3H), 4.77–4.61 (m, 2H), 3.91–3.73 (m, 11H), 2.89–2.81 (m, 2H). ¹³ C NMR (100MHz, CDCl ₃ ) δ162.9, 162.6, 149.6, 148.6, 147.8, 147.8, 147.0, 143.6, 143.1, 130.7, 130.6, 130.4, 129.9, 129.3, 128.8, 126.3, 116.9, 7, 1 111.44,109.7,109.6,109.1,104.9,67.1,56.0,30.0.HRMS(ESI)calcd for C ₂₈ H ₂₆ FN ₃ O ₃ (M+H) ⁺ 472.2031,found 472.2034.

Synthesis of compound 5d:

To a solution of compound 4d (182 mg, 0.6 mmol) in CH ₂ Cl ₂ (5 mL) was added oxalyl chloride (228 mg, 1.8 mmol), then DMF was added dropwise, and the mixture was stirred at room temperature for 2 hours. Concentrate in vacuo, dissolve the residue in THF (10 mL), add 6,7-dimethoxy-1,2,3,4-tetrahydroisoquinoline hydrochloride (138 mg, 0.6 mmol) at 0°C ) and TEA (0.4 mL, 3 mmol) in THF (20 mL), stirred at room temperature for 1 hour, extracted with EA, washed with saturated brine, dried, concentrated, and purified by silica gel column chromatography to give compound 5d (160 mg, 56%) as a pale yellow solid ).

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.77 (d, J=7.9 Hz, 1H), 8.49 (d, J=3.7 Hz, 1H), 7.87 (s, 1H), 7.64-7.54 (m, 3H) ,7.49–7.39(m,2H),7.35(dd,J=8.0,4.7Hz,1H),6.65(d,J=2.5Hz,2H),4.97–4.52(m,2H),3.97–3.79(m , 11H), 3.02-2.81(m, 2H). ¹³ C NMR (100MHz, CDCl ₃ )δ163.74,147.86,147.82,147.16,146.87,146.80,143.30,130.55,129.78,129.19,127.94,123.360,12 117.65,116.85,116.52,110.42,108.06,107.10,98.82,76.25,54.98,54.95,29.46,28.66,27.12.HRMS(ESI)calcd for C ₂₉ H ₂₆ N ₄ O ₃ (M+H) ⁺ 479.2078,found4 2079.

Synthesis of compound SIS3:

To a solution of compound 4e (556 mg, 2 mmol) in THF (20 mL) was added oxalyl chloride (508 mg, 4 mmol), followed by stirring at room temperature for 2 hours. Concentrate in vacuo, dissolve the residue in THF (10 mL), add 6,7-dimethoxy-1,2,3,4-tetrahydroisoquinoline hydrochloride (460 mg, 2 mmol) at 0°C and TEA (606 mg, 6 mmol) in THF (20 mL), stirred at room temperature for 1 hour, extracted with EA, washed with saturated brine, dried, concentrated, and purified by silica gel column chromatography to give yellow solid compound SIS3 (500 mg, 55%).

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.44 (d, J=4.7 Hz, 1H), 8.27 (s, 1H), 7.77 (d, J=15.4 Hz, 1H), 7.59-7.49 (m, 3H) ,7.45(d,J＝7.4Hz,2H),7.28-7.26(m,2H),6.87(d,J＝15.4Hz,1H),6.64(s,2H),4.74-4.70(m,2H), 3.87-3.78 (m, 11H), 2.84 (d, J=20.7Hz, 2H). ¹³ C NMR (100MHz, CDCl ₃ ) δ 166.8, 148.9, 147.8, 144.4, 143.6, 136.0, 130.7, 129.9, 129.4, 128.8, 128.3,126.1,118.5,117.1,113.6,111.2,109.5,109.1,56.0,47.1,44.4,43.7,40.0,29.9,29.2,28.2.HRMS(ESI)calcd for C ₂₈ H ₂₇ N ₃ O ₃ (M+H ) ⁺ 454.2125, found 454.2126.

Synthesis of compound 5f:

Compound SIS3 (220 mg, 0.48 mmol) and Pd/C (22 mg, 10% w/w) were placed in a reaction flask, replaced with hydrogen, dissolved in MeOH (20 mL), and stirred at room temperature overnight. The resulting mixture was filtered and the filtrate was evaporated to give the crude product, which was purified by silica gel column chromatography to give compound 5f as a white solid (120 mg, 54%).

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.34 (d, J=4.3 Hz, 1H), 7.95 (d, J=7.7 Hz, 1H), 7.53-7.40 (m, 5H), 7.11-7.03 (m, 1H), 6.59(d, J＝8.5Hz, 1H), 6.55(s, 0.5H), 6.33(s, 0.5H), 4.59(s, 1H), 4.29(s, 1H), 3.85–3.82(m ,6H),3.75(t,J＝5.9Hz,1H),3.70-3.66(m,3H),3.39(t,J＝5.8Hz,1H),3.14-3.07(m,2H),2.71-2.53( m, 4H). ¹³ C NMR (100MHz, CDCl ₃ )δ171.3, 171.2, 148.1, 147.9, 147.8, 147.6, 147.6, 142.8, 138.2, 138.2, 131.3, 130.4, 128.7, 128.6, 128.5, 126., 9, 125.7.9, 125.4, 124.1, 120.2, 115.5, 115.4, 111.5, 111.1, 110.4, 110.3, 109.4, 108.7, 56.0, 46.9, 43.9, 43.2, 39.6, 35.2, 34.9, 29.5, 28.8, 27.9, 20.4, 20.3 calcd for C ₂₈ H ₂₉ N ₃ O ₃ (M+H) ⁺ 456.2282, found456.2288.

Synthesis of compound 5g:

Compound 4e (200 mg, 0.72 mmol) and oxalyl chloride (254 mg, 2.00 mmol) were placed in a reaction flask, and 10 mL of THF was added. Stir at room temperature for 1 hour. Concentrate under reduced pressure to obtain the intermediate, add 1,2,3,4-tetrahydro-6,7-isoquinolinediol hydrobromide (176 mg, 0.72 mmol) and DIPEA (213 mg, 1.65 mmol), add 10 mL of THF to dissolve . Stir at room temperature for 1.5 hours. The sampling point plate reaction was basically completed, EA was extracted three times, the organic phases were combined, dried, filtered, concentrated, and purified by silica gel column chromatography to obtain compound 5g (130 mg, 42%).

¹ H NMR (400MHz, DMSO-d ₆ ) δ 8.79-8.74 (m, 1H), 8.60 (dd, J=7.6Hz, 1H), 8.43 (d, J=4.0Hz, 1H), 7.67-7.59 ( m, 3H), 7.56–7.54 (m, 2H), 7.48 (d, J=15.4Hz, 1H), 7.33 (s, 1H), 7.05 (d, J=15.4Hz, 1H), 6.64–6.52 (m , 2H), 4.58(d, J＝86.5Hz, 2H), 3.83-3.69(m, 5H), 2.64(d, J＝48.7Hz, 2H). ¹³ C NMR (100MHz, DMSO-d ₆ )δ165. 94,148.69,144.59,144.34,144.23,144.09,135.14,131.22,129.88,129.53,129.22,125.38,124.59,124.23,117.85,115.62,114.18,114.01,113.70,108.75,55.38,44.25,30.01,28.99.HRMS(ESI) calcd for C ₂₆ H ₂₃ N ₃ O ₃ (M+H) ⁺ 426.1812, found 426.1816.

Synthesis of compound 5h:

Compound 4e (200 mg, 0.72 mmol) and oxalyl chloride (254 mg, 2.0 mmol) were placed in a reaction flask, and 10 mL of THF was added. Stir at room temperature for 1 hour. Concentrate under reduced pressure to give the intermediate, add 6,7-bis(2-methoxyethoxy)-1,2,3,4-tetrahydroisoquinoline hydrochloride (210 mg, 0.66 mmol) and DIPEA (213 mg) , 1.65mmol), add 10mL THF to dissolve. Stir at room temperature for 1.5 hours. The sampling point plate reaction was basically completed, EA was extracted three times, the organic phases were combined, dried, filtered, concentrated, and purified by silica gel column chromatography to obtain a yellow solid compound 5h (120 mg, 29%).

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.43 (d, J=4.0 Hz, 1H), 8.24 (s, 1H), 7.76 (d, J=15.4 Hz, 1H), 7.56-7.50 (m, 3H) ,7.45–7.43(m,2H),7.25(s,1H),6.85(d,J＝15.4Hz,1H),6.71(d,J＝3.4Hz,2H),5.29(s,1H),4.69( d, J=12.4Hz, 2H), 4.22–4.04 (m, 4H), 3.90–3.71 (m, 9H), 3.45 (s, 6H), 2.81 (d, J=19.7Hz, 2H). ¹³ C NMR (100MHz, CDCl ₃ )δ166.7,148.8,148.1,147.7,144.4,143.5,135.9,130.7,129.9,129.4,128.8,128.3,127.1,118.6,117.1,115.2,113.6,113.2,109.1 59.2,53.5,47.1,44.3,43.6,40.0,29.9,29.2,28.1.HRMS(ESI)calcd for C ₃₂ H ₃₅ N ₃ O ₅ (M+H) ⁺ 542.2649,found 542.2654.

Synthesis of compound 5i:

To a solution of compound 4f (120 mg, 0.432 mmol) and TEA (0.24 mL) in DMF (3 mL) was added HATU (197 mg, 0.518 mmol), stirred at 0°C for 0.5 h, and added 6,7-dimethoxy-1,2 , 3,4-Tetrahydroisoquinoline hydrochloride (99 mg, 0.432 mmol). Stir at room temperature for 5 hours, sample the plate, the reaction is basically complete, add ice water for dilution, suction filtration, dissolve in ethyl acetate, dry over anhydrous sodium sulfate, filter and concentrate, and purify with silica gel column to obtain the corresponding white solid compound 5i (52 mg, 27% ).

¹ H NMR (400MHz, CDCl ₃ ) δ 7.98 (s, 1H), 7.79 (d, J=15.4Hz, 1H), 7.61-7.27 (m, 10H), 6.91 (d, J=15.4Hz, 1H) , 6.63(s, 2H), 4.72(s, 2H), 3.85(s, 6H), 3.62(s, 3H), 2.85(s, 2H). ¹³ C NMR(100MHz, CDCl ₃ )δ167.3,147.8,144.5 ,138.0,136.8,131.0,130.5,129.1,128.8,125.8,122.9,121.4,120.4,113.0,112.7,111.6,111.3,110.9,110.2,109.6,109.1,56.1,56.1,44.5,4 (ESI)calcd for C ₂₉ H ₂₈ N ₂ O ₃ (M+H) ⁺ 453.2173, found453.2169.

Synthesis of compound 5j:

To a solution of compound 4g (200 mg, 0.72 mmol) and 6,7-dimethoxy-1,2,3,4-tetrahydroisoquinoline hydrochloride (165 mg, 0.72 mmol) in dry DMF (3 mL) DIPEA (1 mL) and HATU (380 mg, 1 mmol) were added, stirred for 16 hours, diluted with water, extracted with EA, washed with saturated brine, dried over anhydrous MgSO ₄ , filtered, concentrated, and purified by silica gel column chromatography to obtain yellow solid product 5j ( 90 mg, 27%).

¹ H NMR (400MHz, DMSO-d ₆ )δ9.48(s,1H),8.65-8.47(m,2H),7.74-7.67(m,3H),7.64-7.56(m,2H),7.46(d , J=15.6Hz, 1H), 7.15(d, J=15.6Hz, 1H), 6.83–6.74(m, 2H), 4.78(s, 1H), 4.60(s, 1H), 3.86(s, 3H) , 3.78–3.67(m, 8H), 2.81(s, 1H), 2.70(s, 1H). ¹³ C NMR (100MHz, CDCl ₃ )δ162.5, 148.3, 148.0, 147.9, 147.8, 133.4, 130.8, 130.4, 129.4 _{( _ _ _} M+H) ⁺ 454.2125, found 454.2129.

Example 2: Synthesis of Compound 9

Compound 6 (208 mg, 1 mmol) and AlCl ₃ (667 mg, 5 mmol) were placed in a reaction flask, replaced with nitrogen, dissolved in 20 mL of DCM, and stirred at room temperature for 1 hour. Methyl 2-chloro-2-oxoacetate (122 mg, 1 mmol) was added, stirred at 0 °C for 0.5 h, then warmed to room temperature and stirred overnight under _N2 protection. Sampling plate, the reaction was basically completed, extracted, extracted with DCM three times, the organic phases were combined, dried, filtered, concentrated, and purified by silica gel column chromatography to obtain yellow solid compound 7 (270 mg, 92%); ¹ H NMR (400 MHz, CDCl ₃ ) δ8.71(d,J＝7.8Hz,1H),8.49(d,J＝4.7Hz,1H),7.63-7.53(m,3H),7.48(d,J＝5.7Hz,2H),7.35(dd , J=7.8, 4.8Hz, 1H), 3.74(s, 3H), 3.24(s, 3H). ¹³ CNMR (100MHz, CDCl ₃ )δ177.5, 162.6, 149.1, 144.0, 137.7, 132.1, 119.8, 119.2, 112.7 , 52.9.

Compound 7 (270 mg, 0.92 mmol) and LiOH (66 mg, 2.76 mmol) were placed in a reaction flask, MeOH (10 mL) and water (5 mL) were added to dissolve, and the mixture was stirred at room temperature overnight. Sampling plate, the reaction was basically completed, the mixture was concentrated under vacuum, extracted with EA (2×30 mL), the organic layers were combined, dried, filtered, concentrated, and purified by silica gel column chromatography to obtain compound 8 (230 mg, 89%) as a white solid; ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.71 (d, J=7.6 Hz, 1H), 8.40 (d, J=4.3 Hz, 1H), 7.65-7.48 (m, 5H), 7.39-7.31 (m, 1H), 3.69(s, 3H). ¹³ C NMR (100MHz, CDCl ₃ )δ182.0, 165.8, 150.5, 147.8, 144.7, 131.4, 130.4, 129.1, 128.5, 119.7, 119.5, 109.2, 30.1.

Compound 8 (230 mg, 0.82 mmol), 6,7-dimethoxy-1,2,3,4-tetrahydroisoquinoline hydrochloride (228 mg, 0.98 mmol), DIPEA (222 mg, 1.72 mmol) and HATU (373 mg, 0.98 mmol) was placed in a reaction flask, DMF (5 mL) was added to dissolve, and the mixture was stirred at room temperature for 8 hours. Sampling point plate, the reaction is basically completed. Concentrated in vacuo, extracted with EA, dried, filtered, concentrated, and purified by silica gel column chromatography to obtain compound 9 (134 mg, 36%) as a white solid; ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.77 (dd, J=18.0, 7.7 Hz) ,1H),8.48(d,J=4.4Hz,1H),7.50-7.42(m,3H),7.39-7.32(m,1H),7.30-7.22(m,2H),6.56(d,J=5.0 Hz, 1H), 6.45(s, 1H), 4.37(s, 1H), 4.01(s, 1H), 3.84(dd, J=19.4, 15.6Hz, 6H), 3.65(d, J=26.2Hz, 3H ), 3.45(t, J＝5.3Hz, 1H), 3.30(t, J＝6.0Hz, 1H), 2.73(t, J＝5.2Hz, 1H), 2.39(t, J＝5.8Hz, 1H). ¹³ C NMR (100 MHz, CDCl ₃ ) δ 187.3, 165.8, 149.3, 148.1, 147.9, 147.8, 144.8, 131.2, 130.2, 128.4, 126.5, 125.5, 123.7, 123.5, 119.4, 111.4, 110.4, 108.8, 9 ,55.95,46.9,43.5,42.5,38.7,29.7,28.5,27.1.HRMS(ESI)calcd for C ₂₇ H ₂₅ N ₃ O ₄ (M+H) ⁺ 456.1918,found 456.1921.

Example 3: Synthesis of Compound 12

NaH (112 mg, 2.8 mmol) was placed in a reaction flask, nitrogen was replaced, 5 mL of anhydrous DMF was added to dissolve, methyl phosphonoacetate diethyl ester (555 mg, 2.2 mmol) was added in an ice-water bath, and the mixture was stirred for 0.5 hours. A solution of compound 10 (451 mg, 2.0 mmol) in DMF was added dropwise and stirred at room temperature for 8 hours. Sampling and spotting, the reaction was basically completed, water was added to quench the reaction, stirred for 2 hours, extracted with ethyl acetate three times, washed with saturated brine, dried, filtered, concentrated, and purified by silica gel column to obtain yellow intermediate n7 (542 mg, 84%).

The intermediate n7 (542 mg, 1.68 mmol) was dissolved in a mixed solution of CH ₂ Cl ₂ (9 mL) and TFA (2 mL), stirred at room temperature overnight, sampled and spotted, and the reaction was basically completed to obtain compound 11 (550 mg, crude product).

To a solution of compound 11 (150 mg, 0.565 mmol) and TEA (0.5 mL, 3.6 mmol) in DMF (3.5 mL) was added HATU (258 mg, 0.678 mmol), stirred at 0°C for 0.5 h, and added 6,7-dimethoxy - 1,2,3,4-Tetrahydroisoquinoline hydrochloride (130 mg, 0.565 mmol). After stirring at room temperature for 5 hours, the spotting reaction was basically completed, diluted with ice water, filtered with suction, and purified by silica gel column to obtain compound 12 (234 mg, 94%); ¹ H NMR (400 MHz, MeOH) δ9.39–9.26 (m, 1H) ,8.89(d,J＝3.5Hz,1H),7.65(t,J＝10.6Hz,3H),7.52(t,J＝6.0Hz,4H),7.14(dd,J＝15.6,9.7Hz,1H) ,6.64–6.48(m,2H),4.54(d,J=16.8Hz,2H),3.71(dd,J=7.6,3.6Hz,1H),3.65(d,J=7.7Hz,6H),3.62– 3.55(m, 1H), 2.67(d, J=5.1Hz, 2H). ¹³ C NMR (100MHz, MeOH) δ 164.9, 164.9, 156.8, 148.1, 148.0, 147.8, 144.8, 138.7, 138.5, 136.2, 136.1, 130.79 ,130.75,129.3,129.2,127.2,127.2,126.7,126.2,124.7,124.5,124.2,122.7,122.6,117.8,113.6,111.6,111.5,109.6,109.4,55.2,55.10,55.5 , 27.4. HRMS(ESI) calcd for C ₂₆ H ₂₄ N ₄ O ₃ (M+H) ⁺ 441.1921, found 441.1921.

Example 4: Synthesis of compounds 13a-13h, compounds 14a-14b

Synthesis of compound 13a:

To a solution of compound 4e (63 mg, 0.227 mmol) and TEA (0.095 mL) in DMF (0.915 mL) was added HATU (104 mg, 0.273 mmol), stirred at 0°C for 0.5 h, and added L-phenylalanine methyl ester hydrochloride (49 mg, 0.227 mmol). Stir at room temperature for 5 hours, sample and spot plate, the reaction is basically complete, add ice water for dilution, suction filtration, dissolve in ethyl acetate, dry over anhydrous sodium sulfate, filter and concentrate, and purify with silica gel column to obtain compound 13a (59 mg, 59%) as a white solid.

¹ H NMR (400MHz, CDCl ₃ ) δ 8.40 (dd, J=4.8, 1.5Hz, 1H), 8.19 (dd, J=7.9, 1.5Hz, 1H), 7.68 (d, J=15.6Hz, 1H) ,7.56–7.49(m,3H),7.44–7.39(m,2H),7.30–7.26(m,2H),7.25–7.17(m,2H),7.11(dd,J=7.9,1.5Hz,2H) ,6.39(d,J＝15.6Hz,1H),6.07(d,J＝7.7Hz,1H),5.01(dt,J＝7.7,5.7Hz,1H),3.74(s,3H),3.73(s, 3H), 3.17 (t, J=6.1Hz, 2H). ¹³ C NMR (100MHz, CDCl ₃ )δ172.4, 166.5, 149.0, 145.2, 143.7, 136.1, 135.3, 130.8, 129.6, 129.5, 129.4, 128.9, 128.7, 128.6,127.1,118.4,117.3,115.8,108.6,53.4,52.4,38.1,30.0.HRMS(ESI)calcd for C ₂₇ H ₂₅ N ₃ O ₃ (M+H) ⁺ 440.1969,found 440.1965.

Synthesis of compound 13b:

To a solution of compound 4e (120 mg, 0.432 mmol) and TEA (0.18 mL) in DMF (1.8 mL) was added HATU (164 mg, 0.432 mmol), stirred at 0°C for 0.5 h, and added D-phenylalanine methyl ester hydrochloride (93 mg, 0.432 mmol). Stir at room temperature for 5 hours, sample and spot plate, the reaction is basically complete, add ice water for dilution, suction filtration, dissolve in ethyl acetate, dry over anhydrous sodium sulfate, filter and concentrate, and purify with silica gel column to obtain compound 13b (85 mg, 44%) as a white solid.

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.40 (dd, J=4.8, 1.4 Hz, 1H), 8.20 (dd, J=7.9, 1.1 Hz, 1H), 7.66 (d, J=15.6 Hz, 1H) ,7.56–7.46(m,3H),7.40(ddd,J=6.3,4.4,2.7Hz,2H),7.23–7.17(m,2H),7.12–7.07(m,2H),6.37(d,J= 15.6Hz, 1H), 4.99 (dt, J=7.7, 5.7Hz, 1H), 3.73 (s, 3H), 3.72 (s, 3H), 3.22–3.10 (m, 2H). ¹³ C NMR (100MHz, CDCl) ₃ ) δ172.4, 166.5, 148.9, 145.2, 143.6, 136.1, 135.3, 130.8, 129.6, 129.6, 129.4, 128.9, 128.7, 128.6, 127.1, 118.5, 117.3, 115.9, 108.6, 38.4, 52 RMS( ESI) calcd for C ₂₇ H ₂₅ N ₃ O ₃ (M+H) ⁺ 440.1969, found 440.1966.

Synthesis of compound 13c:

To a solution of compound 4e (120 mg, 0.432 mmol) and TEA (0.18 mL) in DMF (1.8 mL) was added HATU (164 mg, 0.432 mmol), stirred at 0°C for 0.5 h, and added L-tyrosine methyl ester (84 mg, 0.432 mmol). mmol). Stir at room temperature for 5 hours, sample and spot plate, the reaction is basically complete, add ice water for dilution, suction filtration, dissolve in ethyl acetate, dry over anhydrous sodium sulfate, filter and concentrate, and purify with silica gel column to obtain compound 13c (141 mg, 72%) as a white solid.

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.36 (dd, J=4.8, 1.4 Hz, 1H), 8.15 (dd, J=8.0, 1.4 Hz, 1H), 7.65 (d, J=15.6 Hz, 1H) ,7.49–7.43(m,3H),7.38–7.33(m,2H),7.15(dd,J=7.9,4.8Hz,1H),6.90(d,J=8.5Hz,2H),6.68(d,J ＝8.5Hz, 2H), 6.39(d, J＝15.6Hz, 1H), 6.16(d, J＝7.8Hz, 1H), 4.95(dt, J＝7.7, 5.7Hz, 1H), 3.70(s, 3H ), 3.69(s, 3H), 3.06(qd, J=14.0, 5.7Hz, 2H). ¹³ C NMR (100MHz, CDCl ₃ )δ171.5, 165.7, 154.7, 147.7, 144.3, 142.5, 134.4, 129.6, 129.3, 128.5,128.3,127.8,126.0,117.5,116.3,114.7,114.6,107.4,52.5,51.4,36.2,29.0.HRMS(ESI)calcd for C ₂₇ H ₂₅ N ₃ O ₄ (M+H) ⁺ 456.1918,found 456.1914 .

Synthesis of compound 13d:

Compound 13a (55 mg, 0.125 mmol) and LiOH·H ₂ O (78 mg, 1.859 mmol) were placed in a reaction flask, and a mixed solution of H ₂ O/MeOH (2 mL/2 mL) was added to dissolve, and stirred at room temperature for 4 hours. The pH=5 was adjusted with HCl (1 M) and the white solid compound 13d (49 mg, 92%) was collected by filtration.

HNMR (400MHz, _CDCl3 )δ8.44-7.79(m,3H),7.50(d,J=15.6Hz,1H),7.38(s,3H),7.09(t,J=29.1Hz,9H),6.43 (d, J=6.8Hz, 1H), 6.27 (d, J=15.5Hz, 1H), 4.89 (d, J=4.8Hz, 1H), 3.56 (s, 3H), 3.11 (d, J=32.7Hz) , 2H). ¹³ C NMR (100MHz, CDCl ₃ )δ174.2,167.4,148.6,145.3,143.4,136.2,135.6,130.7,129.6,129.4,129.3,128.9,128.8,128.6,127.0,118.5,115.3,118.5,115.3, ,53.9,37.4,30.0.HRMS(ESI)calcd for C ₂₆ H ₂₃ N ₃ O ₃ (M+H) ⁺ 426.1812,found 426.1810.

Synthesis of compound 13e:

Compound 13b (70 mg, 0.159 mmol) and LiOH·H ₂ O (52 mg, 1.239 mmol) were placed in a reaction flask, and a mixed solution of H ₂ O/MeOH (1 mL/1 mL) was added to dissolve, and stirred at room temperature for 4 hours. The pH=5 was adjusted with HCl (1 M) and the white solid compound 13e (66 mg, 98%) was collected by filtration.

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.39 (dd, J=4.7, 1.2 Hz, 1H), 8.15 (d, J=7.2 Hz, 1H), 7.65 (d, J=15.6 Hz, 1H), 7.57 –7.47(m,3H),7.39(dd,J=7.1,2.2Hz,2H),7.26–7.15(m,6H),6.35(d,J=15.6Hz,1H),6.12(d,J=3.4 Hz, 1H), 4.92 (d, J=5.7Hz, 1H), 3.72 (s, 3H), 3.23 (ddd, J=20.1, 14.0, 5.6Hz, 2H). ¹³ C NMR (100MHz, CDCl ₃ )δ167 .3,148.3,145.3,143.2,136.1,135.4,130.7,129.5,129.4,129.2,129.1,128.8,128.5,127.0,118.6,117.2,115.5,108.5,67.9,37.4,30.1 ₆ H.HRMS (ESIcalc) ₂₃ N ₃ O ₃ (M+H) ⁺ 426.1812, found426.1808.

Synthesis of compound 13f:

Compound 13c (42 mg, 0.092 mmol) and LiOH (16 mg, 0.691 mmol) were placed in a reaction flask, THF/H ₂ O (2 mL/2 mL) mixed solution was added to dissolve, and the mixture was stirred at room temperature for 4 hours. The pH=5 was adjusted with HCl (1 M) and the white solid compound 13f (40 mg, 98%) was collected by filtration.

¹ H NMR (400MHz, MeOH) δ 8.38 (dd, J=8.0, 1.4Hz, 1H), 8.30 (d, J=1.5Hz, 1H), 7.62-7.36 (m, 7H), 7.24 (dd, J =7.9,4.8Hz,1H),7.02(d,J=8.4Hz,2H),6.64(m,J=15.1,12.1Hz,3H),4.58–4.50(m,1H),3.66(s,3H) , 3.02 (ddd, J=20.7, 13.8, 6.0 Hz, 2H). ¹³ C NMR (100 MHz, MeOH) δ 177.2, 167.4, 155.5, 148.5, 144.7, 143.0, 133.0, 130.7, 130.2, 129.6, 129.2, 129.1, 129.0 ,128.6,118.6,117.5,117.1,114.6,108.7,56.4,37.3,28.9.HRMS(ESI)calcd for C ₂₆ H ₂₃ N ₃ O ₄ (M+H) ⁺ 442.1761,found 442.1763.

Synthesis of compound 13g:

To a solution of compound 4e (100 mg, 0.360 mmol) and TEA (0.15 mL) in DMF (1.5 mL) was added HATU (137 mg, 0.360 mmol), stirred at 0°C for 0.5 h, and added 4-chloro-D-phenylalanine methyl Ester hydrochloride (90 mg, 0.360 mmol). Stir at room temperature for 5 hours, sample and spot plate, the reaction is basically complete, add ice water for dilution, suction filtration, dissolve in ethyl acetate, dry over anhydrous sodium sulfate, filter and concentrate, and purify with silica gel column to obtain 13 g (164 mg, 96%) of white solid compound.

¹ H NMR (400MHz, CDCl ₃ ) δ 8.40 (dd, J=4.8, 1.5Hz, 1H), 8.20 (dd, J=7.9, 1.5Hz, 1H), 7.66 (d, J=15.6Hz, 1H) ,7.56–7.46(m,3H),7.41(ddd,J=6.3,4.4,2.6Hz,2H),7.24–7.18(m,3H),7.08–6.96(m,2H),6.37(d,J= 15.6Hz, 1H), 6.00(d, J=7.4Hz, 1H), 4.97(dd, J=13.1, 5.6Hz, 1H), 3.73(s, 3H), 3.72(s, 3H), 3.14(qd, J=13.9, 5.6Hz, 2H). ¹³ C NMR (100MHz, CDCl ₃ )δ172.2, 166.5, 148.9, 145.3, 143.7, 135.5, 134.6, 133.1, 130.8, 130.8, 129.6, 129.6, 128.9, 128.8, 128.7, 118 , 117.4, 115.6, 108.5, 53.3, 52.5, 37.5, 30.0. HRMS(ESI) calcd for C ₂₇ H ₂₄ ClN ₃ O ₃ (M+H) ⁺ 474.1579, found 474.1577.

Synthesis of compound 13h:

To a solution of compound 4e (93 mg, 0.334 mmol) and TEA (0.23 mL) in DMF (2.5 mL) was added HATU (127 mg, 0.334 mmol), stirred at 0°C for 0.5 h, and added n15 (78 mg, 0.334 mmol). Stir at room temperature for 5 hours, sample and spot plate, the reaction is basically complete, add ice water for dilution, suction filtration, dissolve in ethyl acetate, dry over anhydrous sodium sulfate, filter and concentrate, and purify with silica gel column to obtain white solid compound 13h (45mg, 27%).

¹ H NMR (400MHz, MeOH) δ 8.97 (dd, J=8.0, 1.2Hz, 1H), 8.56 (dd, J=5.8, 1.2Hz, 1H), 7.71 (dd, J=8.0, 5.8Hz, 1H) ),7.68–7.61(m,3H),7.54(ddd,J=5.5,2.9,1.6Hz,2H),7.46(d,J=16.0Hz,1H),7.32–7.16(m,6H),6.90( d, J=16.0Hz, 1H), 4.54 (dd, J=8.5, 6.7Hz, 1H), 3.82 (s, 3H), 3.63 (ddd, J=15.1, 7.1, 4.8Hz, 1H), 3.41–3.32 (m, 2H), 3.27–3.12 (m, 3H), 3.03 (dd, J=13.8, 8.5Hz, 1H), 2.87 (d, J=0.6Hz, 6H). ¹³ C NMR (100MHz, MeOH) δ174 .9, 169.0, 148.3, 143.5, 138.2, 137.1, 133.4, 133.3, 132.1, 131.7, 130.3, 129.7, 129.2, 128.0, 123.9, 120.6, 118.4, 111.6, 58.4, 57.3, 44.0, 3.5. (ESI)calcd for C ₃₀ H ₃₃ N ₅ O ₂ (M+H) ⁺ 496.2707, found 496.2706.

Synthesis of compound 14a:

To a solution of compound 4e (102.5 mg, 0.369 mmol) and TEA (0.155 mL) in DMF (2 mL) was added HATU (140 mg, 0.369 mmol), stirred at 0°C for 0.5 h, added 2-(1-piperazinyl)pyrimidine ( 60.51 mg, 0.367 mmol). Stir at room temperature for 5 hours, sample and spot plate, the reaction is basically completed, add ice water for dilution, suction filtration, dissolve in ethyl acetate, dry over anhydrous sodium sulfate, filter and concentrate, and purify with silica gel column to obtain compound 14a (52 mg, 33%) as a white solid.

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.40 (dd, J=4.7, 1.4 Hz, 1H), 8.31 (d, J=4.8 Hz, 2H), 8.19 (dd, J=7.9, 1.4 Hz, 1H) ,7.76(d,J＝15.3Hz,1H),7.58–7.34(m,5H),7.22(dd,J＝7.8,4.7Hz,1H),6.77(d,J＝15.4Hz,1H),6.51( t, J=4.8Hz, 1H), 3.86(s, 4H), 3.73(s, 7H). ¹³ CNMR (100MHz, CDCl ₃ ) δ 166.7, 161.6, 157.9, 148.9, 144.6, 143.7, 136.5, 130.8, 129.9, 129.5,128.9,128.3,118.6,117.3,112.9,110.5,109.1,43.8,29.9,29.4.HRMS(ES)calcdfor C ₂₅ H ₂₄ N ₆ O(M+H) ⁺ 425.2084,found 425.2078.

Synthesis of compound 14b:

To a solution of compound 4e (120 mg, 0.432 mmol) and TEA (0.18 mL) in DMF (1.8 mL) was added HATU (164 mg, 0.432 mmol), stirred at 0°C for 0.5 h, added 1-(4-pyridyl)piperazine ( 70.5 mg, 0.443 mmol). Stir at room temperature for 5 hours, sample and spot plate, the reaction is basically complete, add ice water for dilution, suction filtration, dissolve in ethyl acetate, dry over anhydrous sodium sulfate, filter and concentrate, and purify with silica gel column to obtain compound 14b (163 mg, 89%) as a white solid.

¹ H NMR (400MHz, DMSO-d ₆ ) δ 8.58 (dd, J=7.9, 1.4Hz, 1H), 8.44 (dd, J=4.7, 1.4Hz, 1H), 8.26 (d, J=7.2Hz, 2H), 7.66–7.59 (m, 3H), 7.57–7.54 (m, 2H), 7.51 (d, J=15.4Hz, 1H), 7.33 (dd, J=7.9, 4.7Hz, 1H), 7.09 (d , J=7.4Hz, 2H), 7.04(d, J=15.4Hz, 1H), 3.71(s, 3H), 3.70(s, 8H). ¹³ C NMR(100MHz, DMSO-d ₆ )δ166.1,156.4, 148.8,145.1,144.3,143.2,135.8,131.3,130.0,129.8,129.6,129.3,117.9,117.9,113.3,108.8,108.2,49.3,30.1,27.4.HRMS(ESI)calcd for C ₂₆ H ₂₅ N ₅ O( M+H) ⁺ 424.2132, found 424.2128.

Example 5: Synthesis of Compounds 16a-16d

Synthesis of Intermediate 15a:

To a solution of 3c (4.3 g, 18.2 mmol) in THF (30 mL) at 0°C was added LDA (18.2 mL, 36.4 mmol) in THF dropwise. The temperature was raised to room temperature and stirred for 1 hour, 1-acetylpyrrolidin-2-one (9.25 g, 72.9 mmol) was added, and the mixture was stirred at 45° C. for 48 hours. The reaction of the sampling point plate was basically completed, and was quenched by adding saturated NaHCO ₃ . It was diluted with NH ₄ Cl (3 mL) and water (50 mL), extracted with EA, washed with saturated brine (50 mL), dried over anhydrous Na ₂ SO ₄ , concentrated, and purified by silica gel column chromatography to obtain compound 15a (2.3 mg, 42%) as a white solid. ); ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.42 (dd, J=4.7, 1.4 Hz, 1H), 8.03 (dd, J=7.9, 1.4 Hz, 1H), 7.61-7.37 (m, 6H), 7.18(dd,J=7.9,4.7Hz,1H),6.86(s,1H),3.82(s,3H),3.32(t,J=6.6Hz,2H),2.47–2.27(m,2H). ¹³ C NMR (100MHz, CDCl ₃ ) δ 172.9, 148.4, 143.4, 141.3, 131.0, 130.6, 128.9, 128.8, 128.0, 123.1, 119.7, 116.5, 108.5, 39.7, 29.9, 26.9.

Synthesis of compound 16a:

To a solution of compound 15a (450 mg, 1.5 mmol) in THF (30 mL) was added NaH (80 mg, 2 mmol) in portions and stirred at 0°C for 0.5 h. 4-(bromomethyl)-1,2-dimethoxybenzene (416 mg, 1.8 mmol) was added, stirred at room temperature for 5 hours, the sampling and spot plate reaction was basically completed, extracted with EA, washed with saturated brine, anhydrous Na ₂ Dry over SO ₄ , concentrated, and purified by silica gel column chromatography to give compound 16a (210 mg, 31%) as a pale yellow solid.

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.40 (d, J=4.7 Hz, 1H), 8.02 (d, J=7.9 Hz, 1H), 7.55 (t, J=2.7 Hz, 1H), 7.53-7.40 (m, 5H), 7.15(dd, J=7.7, 4.8Hz, 1H), 6.82–6.77(m, 3H), 4.47(s, 2H), 3.86(s, 6H), 3.79(s, 3H), 3.12(t, J=6.6Hz, 2H), 2.27-2.18(m, 2H). ¹³ C NMR (100 MHz, CDCl ₃ ) δ 169.3, 149.2, 148.5, 148.4, 143.4, 141.1, 131.1, 130.6, 129.7, 129.2, 128.9,128.8,128.0,122.9,120.8,119.7,116.4,111.5,111.0,108.5,56.0,55.9,47.1,43.9,29.9,24.6.HRMS(ESI)calcd for C ₂₈ H ₂₇ N ₃ O ₃ (M+H ) ⁺ 454.2125, found 454.2127.

Synthesis of compound 16b:

Using compound 3c (300 mg, 1.27 mmol) and 1-acetylpiperidin-2-one (716 mg, 5.08 mmol) as raw materials, referring to the synthetic method and procedure of compound 16a, a pale yellow solid compound 16b (60 mg, 13%) was prepared ).

¹ H NMR (400MHz, CDCl ₃ ) δ 8.36(d, J=0.4Hz, 1H), 7.94(s, 1H), 7.57-7.52(m, 6H), 7.20(s, 1H), 6.93-6.78( m, 3H), 4.52(s, 2H), 3.75(d, J=0.8Hz, 9H), 3.27(s, 2H), 2.42(s, 2H), 1.66(s, 2H). ¹³ C NMR(100MHz) , CDCl ₃ )δ165.3,149.2,148.4,143.3,140.8,128.2,120.6,116.3,111.5,111.0,108.6,56.0,55.9,50.8,47.2,30.0,27.4,23.1.HRMS(ESI)calcd for C ₂₉ H ₂₉ N ₃ O ₃ (M+H) ⁺ 468.2282, found 468.2285.

Synthesis of compound 16c:

Using compound 15a (100 mg, 0.330 mmol) and 2-chloromethyl-3,5-dimethyl-4-methoxypyridine (50 mg, 0.224 mmol) as raw materials, referring to the synthetic method and procedure of compound 16a, the obtained Light yellow solid compound 16c (19 mg, 12%).

¹ H NMR(400MHz,MeOH)δ8.62(d,J=7.7Hz,1H),8.51(d,J=5.4Hz,1H),8.43(s,1H),7.65-7.53(m,6H), 7.42(s, 1H), 4.81(s, 2H), 4.14(s, 3H), 3.86(s, 3H), 3.44(t, J＝5.4Hz, 2H), 2.58(s, 2H), 2.47(s ,3H),2.38(s,3H). ¹³ C NMR(100MHz,MeOH)δ171.6,170.8,149.0,143.9,142.6,140.6,136.6,134.4,131.1,132.6,129.9,129.2,129.1,129.0,123.1,122. ,116.4,109.7,60.9,45.3,43.6,30.4,24.7,13.2,10.2.HRMS(ESI)calcd forC ₂₈ H ₂₈ N ₄ O ₂ (M+H) ⁺ 453.2285,found 453.2281.

Synthesis of compound 16d:

Using compound 15a (500mg, 1.65mmol) and 2-methanesulfonyloxymethyl-3,5,6-trimethylpyrazine n16 (891mg, 3.869mmol) as raw materials, referring to the synthetic method and steps of compound 16a, prepared Compound 16d (76 mg, 11%) was obtained as a pale yellow solid.

¹ H NMR (400MHz, CDCl3) δ 8.41 (dd, J=4.7, 1.5Hz, 1H), 8.03 (dd, J=7.9, 1.5Hz, 1H), 7.56 (t, J=2.7Hz, 1H), 7.54–7.41(m, 5H), 7.17(dd, J=7.9, 4.7Hz, 1H), 4.68(s, 2H), 3.81(s, 3H), 3.21(t, J=6.6Hz, 2H), 2.54 (s, 3H), 2.50 (s, 3H), 2.47 (s, 3H), 2.26 (td, J=6.6, 2.7 Hz, 2H).; ¹³ C NMR (100 MHz, CDCl3) δ 169.3, 150.3, 148.9, 148.5 , _{145.8,143.4,141.1,131.1,130.6,129.2,128.9,128.8,128.0,123.2,119.7,116.5,108.5,46.5,44.6,29.9,24.8,21.6,21.5,20.7} ; ₂₇ N ₅ O(M+H) ⁺ 438.2288, found438.2295.

Example 6: Pharmacological effects of azaindoles or their salts

Luciferase reporter gene screening experiment, the experimental steps are as follows: a) Infect A549 cells with adenovirus carrying the SMAD3 reporter gene for 24 hours. b) Cells were pretreated with blank or test compound (5 μM and 10 μM) for 2 h, and then stimulated with 0.1 ng/mL TGF-β for 24 h. c) Lysing the cells, adding the luciferase substrate to the cell lysate, testing the luciferase activity, and comparing the luciferase activity of the test compound with the luciferase activity of the blank group to obtain the inhibition of SMAD3 phosphorylation level. The results are shown in Table 1.

Table 1 Inhibitory activity of azaindoles on SMAD3 phosphorylation (inhibition rate %, μM)

The above activity test results show that the compounds of the present invention show inhibitory activity on the test cells, and can effectively inhibit the phosphorylation of SMAD3 protein.

Example 7: Pharmaceutical properties of azaindoles or their salts

At 25°C, compound SIS3, compound 16d, compound 13h, compound 5b or their HCl salts (prepared by dissolving the compound in 4 equivalents of HCl in methanol, then spin-drying the solvent, and vacuum-drying) were added to pure water or in phosphate buffer (50 mM, pH 7.4). After shaking and centrifugation, the supernatant was taken to measure the concentration of compound SIS3, compound 16d, compound 13h, or compound 5b in the solution to calculate the corresponding solubility. Detection method: HPLC (Agilent 1260Infinity) was used for detection, the chromatographic column was Shimadzu-GL WondaSil C18-WR, the mobile phase was acetonitrile-water-TFA (68/32/1‰), the detection wavelength was 254 nm, and the injection volume was 10 μL.

The results are shown in Table 2: the azaindole compounds of the present invention have good water solubility, and the water solubility of compound 16d, compound 13h and compound 5b is much better than that of the existing SMAD3 small molecule inhibitor compound SIS3 and compound 16d The solubility of the hydrochloride of compound 13h and compound 5b in water is 667 times, 163 times and 723 times that of compound SIS3·HCl in water, respectively.

Table 2 Solubility test results

Example 8: Compound 16d selectively inhibits SMAD3 activation

A549 cells were pretreated without or with drugs (1 μM and 5 μM) for 2 h, then stimulated with 1 ng/mL TGF-β for 30 min, cells were lysed and subjected to Western blot analysis, with phospho-SMAD3 or phospho-SMAD2 antibodies and corresponding secondary antibodies ( After incubation with LI-COR IRDye 800 labeled with LI-COR IRDye 800 for signal detection, the membrane was washed with stripping buffer (25mM Glycine-HCl, pH2, 1% SDS) for 30 minutes, and then re-detected with SMAD3 or SMAD2 primary antibody, the results are shown in Figure 1 Shown: Compound 16d selectively inhibits SMAD3 protein activation, but not SMAD2 protein activation.

Example 9: Anticancer activity of water-soluble compound 16d in LLC transplanted tumor model in B1 mice

C57BL/6 mice were subcutaneously inoculated with 2×10 ⁶ LLC cells (CRL-1642, ATCC). When the tumor size reached 50 mm, the mice were randomly divided into ⁴ groups of 6 mice and received compound 16d (0, 1.25, 2.5 or 5.0 μg/g/day, prepared with normal saline for injection, intraperitoneal injection) 20 days of treatment. Tumor volume was measured every 5 days, and tumor weight was measured at the end of the experiment. As shown in Figure 2, treatment with water-soluble compound 16d in mice significantly reduced LLC tumor volume and weight in a dose-dependent manner.

Next, the mechanism by which compound 16d attenuates lung cancer progression in mice was investigated by pathological sections of mice. Results As shown in Figure 3, Compound 16d-treated mice increased the number of tumor-infiltrating NK cells in a dose-dependent manner, with a significant increase in 16-day treatment at doses of 2.5 or 5.0 μg/g compared to control mice accumulation of NK cells, suggesting an enhanced antitumor immune response in mice treated with compound 16d.

These results suggest that compound 16d can significantly inhibit cancer progression by enhancing NK cell-mediated anticancer immunity in LLC model mice.

The technical features of the above-described embodiments can be combined arbitrarily. For the sake of brevity, all possible combinations of the technical features in the following embodiments are not described. However, as long as there is no contradiction between the combinations of these technical features, All should be regarded as the scope described in this specification.

The above-mentioned embodiments only represent several embodiments of the present invention, and the descriptions thereof are specific and detailed, but should not be construed as a limitation on the scope of the patent of the present invention. It should be pointed out that for those of ordinary skill in the art, without departing from the concept of the present invention, several modifications and improvements can also be made, which all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention should be subject to the appended claims.

Claims (18)

1. Azaindole compounds having the structure represented by formula (I) or a pharmaceutically acceptable salt thereof or a stereoisomer thereof or a prodrug molecule thereof:

in: D is selected from: C, N; and when D is N, R is absent _; A, B are independently selected from: N, CR ₃ ; R ₁ is selected from: H, C ₁ -C ₆ alkyl, C ₆ -C ₁₀ aryl, C ₃ -C ₈ cycloalkyl, halogen, CN; R ₂ is selected from: H, C ₁ -C ₆ alkyl; Each R ₃ is independently selected from: H, C ₁ -C ₆ alkyl, halogen, CN; R ₄ is selected from:

R ₅ , each R _5a and R _5b are each independently selected from: H, halogen, CN, C ₁ -C ₆ alkyl, C ₃ -C ₈ cycloalkyl; Each R _is independently selected from:

R ₇ is selected from: H, C ₁ -C ₆ alkyl, C ₆ -C ₁₀ aryl substituted with one or more R ₁₃ , 6-10-membered heteroaryl substituted with one or more R ₁₃ ; Each R ₈ is independently selected from: H, halogen, CN, C ₁ -C ₆ alkyl, C ₃ -C ₈ cycloalkyl, hydroxy, C ₁ -C ₆ alkoxy, C ₁ -C ₆ alkoxy substituted C ₁ -C ₆ alkoxy; Each R ₉ is independently selected from: H, halogen, CN, C ₁ -C ₆ alkyl, C ₃ -C ₈ cycloalkyl, hydroxy, C ₁ -C ₆ alkoxy, C ₁ -C ₆ alkoxy substituted C ₁ -C ₆ alkoxy; R ₁₀ is selected from: H, C ₁ -C ₆ alkyl; R ₁₁ is selected from: hydroxyl, C ₁ -C ₆ alkoxy, -N(R ₁₄ )(R ₁₅ ); R ₁₂ is selected from: H, C ₁ -C ₆ alkyl, one or more R ₁₃ substituted C ₆ -C ₁₀ aryl, one or more R ₁₃ substituted 6-10-membered heteroaryl; Each R ₁₃ is independently selected from: H, halogen, CN, C ₁ -C ₆ alkyl, C ₃ -C ₈ cycloalkyl, hydroxy, C ₁ -C ₆ alkoxy, C ₁ -C ₆ alkoxy substituted C ₁ -C ₆ alkoxy; R ₁₄ and R ₁₅ are each independently selected from: H, C ₁ -C ₆ alkyl, amino substituted C ₁ -C ₆ alkyl, C ₁ -C ₆ alkylamino substituted C ₁ -C ₆ alkyl; n is selected from: 1, 2, 3, 4; m is selected from: 0, 1, 2, 3, 4; p is selected from: an integer between 0-7; q is selected from: 1, 2, 3, 4; y is selected from: 1, 2, 3, 4, 5; And, when D is C, A is N, B is CR ₃ , R ₁ is phenyl, and R ₂ is

, R ₈ is not C ₁ -C ₆ alkoxy. 2. the azaindole compound according to claim 1 or its pharmaceutically acceptable salt or its stereoisomer or its prodrug molecule, is characterized in that, D is C, A is N, B is CR ₄ . 3. the azaindole compound according to claim 1 or its pharmaceutically acceptable salt or its stereoisomer or its prodrug molecule, is characterized in that, described azaindole compound has formula ( II), formula (IV), formula (V) or the structure shown in formula (VI):

4 . The azaindole compound according to claim 3 or a pharmaceutically acceptable salt thereof or a stereoisomer thereof or a prodrug molecule thereof, wherein R ₅ is H. 5 . 5. the azaindole compound according to claim 3 or its pharmaceutically acceptable salt or its stereoisomer or its prodrug molecule is characterized in that, n is selected from: 1, 2; m is selected from : 1 or 2. 6. the azaindole compound according to claim 3 or its pharmaceutically acceptable salt or its stereoisomer or its prodrug molecule is characterized in that, R ₇ is selected from: one or more R ₁₃ Substituted phenyl, six-membered nitrogen-containing heteroaryl substituted with one or more R ₁₃ . 7. the azaindole compound according to claim 6 or its pharmaceutically acceptable salt or its stereoisomer or its prodrug molecule is characterized in that, each R ₁₃ is independently selected from: H, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy. 8. the azaindole compound according to claim 6 or its pharmaceutically acceptable salt or its stereoisomer or its prodrug molecule is characterized in that, R ₇ is selected from:

9. the azaindole compound according to claim ₃ or its pharmaceutically acceptable salt or its stereoisomer or its prodrug molecule is characterized in that, each R is independently selected from: H, hydroxy, C ₁ -C ₃ alkoxy, C ₁ -C ₃ alkoxy substituted C ₁ -C ₃ alkoxy; more preferably, each R ₈ is independently selected from: hydroxy, methoxy, methyl oxy-substituted ethoxy; more preferably, both R ₈ are hydroxy. 10. the azaindole compound according to claim 3 or its pharmaceutically acceptable salt or its stereoisomer or its prodrug molecule is characterized in that, R ₉ is selected from: H, halogen, hydroxyl; R ₁₀ is selected from: H, C ₁ -C ₃ alkyl; R ₁₁ is selected from: C ₁ -C ₆ alkoxy. 11. the azaindole compound according to claim 3 or its pharmaceutically acceptable salt or its stereoisomer or its prodrug molecule is characterized in that, R ₁₂ is selected from: one or more R ₁₃ Substituted C ₆ -C ₁₀ heteroaryl; each R ₁₃ is independently selected from: H, C ₁ -C ₆ alkyl; More preferably, R ₁₂ is selected from:

12. the azaindole compound according to any one of claim 1-11 or its pharmaceutically acceptable salt or its stereoisomer or its prodrug molecule, it is characterized in that, R _5a and R _5b respectively independently selected from: H, halogen; and/or, R ₁ is selected from: H, C ₁ -C ₃ alkyl, C ₆ -C ₁₀ aryl, C ₃ -C ₆ cycloalkyl; and/or, R ₂ is selected from: C ₁ -C ₃ alkyl. 13. The azaindoles according to claim 12 or a pharmaceutically acceptable salt thereof or a stereoisomer thereof or a prodrug molecule thereof, wherein R _5b is H, and R _5a is F; and /or, R ₁ is selected from: H, phenyl, cyclopropyl; and/or, _R3 is H. 14. The azaindole compound according to claim 1 or its pharmaceutically acceptable salt or its stereoisomer or its prodrug molecule is characterized in that, it is selected from the following compounds:

15. Use of the azaindole compound according to any one of claims 1 to 14, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a prodrug molecule thereof, in the preparation of an SMAD3 inhibitor. 16. Use of the azaindole compounds according to any one of claims 1-14 or their pharmaceutically acceptable salts or their stereoisomers or their prodrug molecules in the preparation of a medicament for preventing and/or treating tumors application. The application according to claim 16, wherein the tumor is: lung cancer, melanoma, breast cancer, liver cancer. 18. A pharmaceutical composition for preventing and/or treating tumors, comprising an active ingredient and a pharmaceutically acceptable carrier, wherein the active ingredient comprises the azaindoline according to any one of claims 1-14 Indoles or their pharmaceutically acceptable salts or their stereoisomers or their prodrug molecules.

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