AKR1C3-targeted small-molecule coupling medicine as well as preparation method and application of AKR1C3-targeted small-molecule coupling medicine

Abstract

The invention discloses an AKR1C3-targeted micromolecule coupling drug and a preparation method and application thereof, the AKR1C3-targeted micromolecule coupling drug comprises a compound shown as a formula (I) or pharmaceutically acceptable salt thereof, the structure of the compound has good inhibitory activity and selectivity on AKR1C3, cytotoxic molecules can be specifically released in a tumor microenvironment, and tumor cell proliferation is effectively inhibited. The compound is applied to treatment of AKR1C3 overexpression-related diseases, and particularly has a relatively good application prospect in the anti-tumor field.

Description

Technical Field

[0001] This application relates to a compound, its preparation method and application, specifically to a small molecule conjugate drug targeting AKR1C3, its preparation method and application. Background Technology

[0002] The aldehyde-ketone reductase (AKR) superfamily comprises 16 distinct enzyme families that catalyze the stereospecific reduction of NADP(H)-dependent aldehydes and ketones. Among them, AKR1C3 (type 5 17β-hydroxysteroid dehydrogenase / prostaglandin F synthase) plays a crucial role in hormone metabolism and biosynthesis, regulating the activity of androgens, estrogens, progestins, and prostaglandins by reducing 3-keto, 17-keto, and 20-ketosteroids. AKR1C3 is overexpressed in various malignant tumors (including hepatocellular carcinoma (HCC), T-cell acute lymphoblastic leukemia (T-ALL), breast cancer, prostate cancer, and endometrial cancer) as well as non-malignant diseases (such as polycystic ovary syndrome (PCOS) and endometriosis). AKR1C3 inhibitors have shown potential in reversing chemotherapy resistance. For example, in acute myeloid leukemia (AML) and T-ALL models, combination therapy with etoposide, cytarabine, or daunorubicin significantly enhances antitumor efficacy.

[0003] However, monotherapy with AKR1C3 inhibitors has shown limited therapeutic potential. To address this issue, a prodrug strategy linking AKR1C3 inhibitors with cytotoxic agents has emerged. PR-104 is one such targeted agent.

[0004] The hypoxia-activated prodrug of AKR1C3 is rapidly hydrolyzed in vivo to PR-104A. Subsequently, in the hypoxic tumor microenvironment, AKR1C3-mediated reduction generates the cytotoxic metabolites PR-104H and PR-104M. Despite showing efficacy in preclinical studies, development of PR-104 has been terminated due to its limited tumor tissue targeting in clinical trials, leading to severe off-target toxicities (including myelosuppression and neutropenia).

[0005] AST-3424 is a nitrogen mustard prodrug selectively activated by AKR1C3 overexpression, which entered Phase II clinical trials in 2021. Upon enzymatic reduction, this prodrug releases a bifunctional alkylated aziridine that can induce DNA cross-linking, exhibiting potent activity against aggressive / chemoresistant T-ALL and HCC.

[0006] The rapid development of small molecule conjugates targeting AKR1C3 proves the therapeutic potential of the target, however, current research mainly focuses on the nitroreduction function of AKR1C3, the mechanism is single, and no drug is approved for marketing, therefore, developing new, powerful and specific small molecule conjugates targeting AKR1C3 has good application prospect. SUMMARY

[0007] The application discloses a small molecule conjugate drug targeting AKR1C3 and a preparation method and application thereof.

[0008] The small molecule conjugate drug comprises a compound as shown in the formula (I) or a pharmaceutically acceptable salt thereof.

[0009]

[0010] wherein:

[0011] R1, R2, R3 and R4 are independently selected from hydrogen, halogen, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 haloalkoxy, C1-C4 alkoxy, nitro, amino, dimethylamino, hydroxyl and cyano;

[0012] n is selected from 0, 1 and 2;

[0013] X is selected from

[0014]

[0015] L1 is selected from C1-C6 carbon chain and C1-C8 oxygen-containing carbon chain;

[0016] L2 is selected from any one of the following structures:

[0017]

[0018] H2N-D is selected from an antitumor drug containing an aromatic amine structure and an antitumor drug containing a fatty amine structure.

[0019] HN-D is an imino group obtained by removing one hydrogen atom from the amino group in H2N-D.

[0020] The small molecule conjugate drug,

[0021] R1, R2, R3, R4are independently selected from hydrogen, fluorine, chlorine, bromine, iodine, methyl, ethyl, propyl, isopropyl, C1-C4haloalkyl, C1-C4haloalkoxy, methoxy, ethoxy, nitro, amino, dimethylamino, hydroxyl, cyano;

[0022] n is selected from 0, 1, 2;

[0023] L1is selected from any one of the following structures:

[0024]

[0025] -HN-D is selected from any one of the following structures:

[0026]

[0027] The small molecule conjugated drug, the compound is selected from any one of the following compounds:

[0028]

[0029]

[0030]

[0031] The small molecule conjugated drug, the pharmaceutically acceptable salt is the salt of the compound and any one of the following acids:

[0032] hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, carbonic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, citric acid, tartaric acid, malic acid, lactic acid, pyruvic acid, acetic acid, maleic acid, succinic acid, fumaric acid, salicylic acid, phenylacetic acid, mandelic acid, ferulic acid.

[0033] The preparation method of the small molecule conjugated drug is selected from any one of the following methods:

[0034] Method one:

[0035]

[0036] Compounds 1 and 2 undergo nucleophilic substitution reaction to obtain intermediate 3, and then hydrolysis to obtain carboxylic acid intermediate 4, 4 and compound 5 are subjected to amide condensation to obtain intermediate 6, and further hydrolysis to obtain intermediate 7;

[0037] Compounds 8 and 9 are subjected to amide condensation to obtain intermediate 10, 10 is removed from the Boc protecting group to obtain intermediate 11, 11 and compound 12 are subjected to amide condensation to obtain intermediate 13, and 13 is removed from the Boc protecting group to obtain intermediate 14;

[0038] After amide condensation of intermediate 7 and intermediate 14, the compound of formula I is obtained;

[0039] Method two:

[0040]

[0041] amide condensation of intermediate 7 and intermediate 11 obtained in the same way as in method one to obtain the compound of formula I;

[0042] wherein R1, R2, R3, R4, L1, L2, n are defined as above.

[0043] The preparation method of the small molecule conjugated drug, and the pharmaceutically acceptable salt thereof is to form a salt of the corresponding acid and compound I, namely.

[0044] The pharmaceutical composition comprises the small molecule conjugated drug and a pharmaceutically acceptable carrier.

[0045] The small molecule conjugated drug or the pharmaceutical composition is used for preparing a drug for preventing or treating cancer.

[0046] The application, the prevention or treatment of cancer, includes preventing cancer, treating cancer or reversing tumor drug resistance.

[0047] The application, the cancer includes liver cancer, breast cancer, lung cancer. Preferably, the lung cancer includes non-small cell lung cancer.

[0048] When H2N-D is 3-(4-aminobenzyl)-5-fluoropyrimidine-2,4(1H,3H)-dione (intermediate F1), the preparation method is as follows:

[0049]

[0050] (1) Synthesis of 5-fluoro-3-(4-nitrobenzyl)-1-(tetrahydrofuran-2-yl)pyrimidine-2,4(1H,3H)-dione (intermediate F1-1)

[0051] Tegafur (300 mg, 1.50 mmol) and potassium carbonate (518 mg, 3.75 mmol) were dissolved in 6 ml of DMF, followed by the addition of 4-nitrobenzyl bromide (324 mg, 1.50 mmol), stirring at room temperature for 30 min, quenching the reaction by adding 10 ml of water, extracting with ethyl acetate, washing with saturated NaCl solution, drying over anhydrous sodium sulfate, and removing the ethyl acetate under vacuum. After purification by column chromatography (PE:EA = 3:1, v / v), vacuum drying was performed to obtain a white solid (intermediate F1-1, 407 mg, yield 81%) 1H NMR (300 MHz, DMSO-d6): δ 8.37 - 8.27 (m, 2H), 8.17 (d, J = 6.6 Hz, 1H), 7.73 - 7.68 (m, 2H), 6.09 (t, J = 6.7, 3.3, 1.6 Hz, 1H), 5.24 (s, 2H), 4.42 (dt, J = 8.1, 6.1 Hz, 1H), 3.96 (q, J = 7.5 Hz, 1H), 2.38 (dt, J = 14.4, 6.9 Hz, 1H), 2.19 (dtd, J = 12.6, 6.0, 3.1 Hz, 1H), 2.08 (q, J = 6.9 Hz, 2H).

[0052] (2) Synthesis of 3-(4-aminobenzyl)-5-fluoropyrimidine-2,4(lH,3H)-dione (Intermediate Fl)

[0053] Intermediate Fl-1 (300 mg, 0.89 mmol) and anhydrous stannous chloride (424 mg, 2.24 mmol) were dissolved in 10 ml of methanol solution, then 4N HC1 solution was added dropwise until pH = 1-2, warmed to 70°C, and reacted for 3h, with TLC monitoring of the reaction (developing agent PE:EA = 1:5, v / v), the raw material point disappeared, and the reaction was completed. Saturated sodium bicarbonate solution was slowly added dropwise in an ice bath, suction filtered, the filtrate was extracted with ethyl acetate, washed with saturated NaCl solution, dried over anhydrous sodium sulfate, and the ethyl acetate was removed in vacuo. After purification by column chromatography (PE:EA = 1:1, v / v), vacuum drying was performed to obtain a white solid (Intermediate 9a, 135 mg, yield 64%) 1 H NMR (600 MHz, DMSO-d6): δ 11.06 (s, 1H), 7.81 (d, J = 5.5 Hz, 1H), 7.02 - 6.96 (m, 2H), 6.50 - 6.42 (m, 2H), 4.99 (s, 2H), 4.76 (s, 2H).

[0054] When H2N-D is 1-(4-aminobenzyl)-5-fluoropyrimidine-2,4(lH,3H)-dione (Intermediate F2), it is prepared as follows:

[0055]

[0056] (1) Synthesis of 5-fluoro-l-(4-nitrobenzyl)pyrimidine-2,4(lH,3H)-dione (Intermediate F2-1)

[0057] Referring to the synthesis of Intermediate Fl, replace tegafur in the synthesis route of Intermediate Fl with 5-fluorouracil to obtain a white solid (Intermediate F2-1) 1H NMR (400 MHz, DMSO-d6): δ 11.29 (s, 1H), 8.20 (d, J = 1.3 Hz, 1H), 8.18 (d, J = 1.3 Hz, 1H), 8.06 (d, J = 20.2 Hz, 1H), 7.71 (t, J = 1.1 Hz, 1H), 7.69 (q, J = 1.1 Hz, 1H), 5.08 (t, J = 1.1 Hz, 2H).

[0058] (2) Synthesis of 1-(4-aminobenzyl)-5-fluoropyrimidine-2,4(lH,3H)-dione (Intermediate F2)

[0059] Referring to the synthesis of Intermediate Fl, Intermediate Fl-1 in the synthesis route of Intermediate Fl is replaced by Intermediate F2-1 to obtain a white solid (Intermediate F2) 1 H NMR (400 MHz, DMSO-d6): δ 11.29 (s, 1H), 8.20 (d, J = 1.3 Hz, 1H), 8.18 (d, J = 1.3 Hz, 1H), 8.06 (d, J = 20.2 Hz, 1H), 7.71 (t, J = 1.1 Hz, 1H), 7.69 (q, J = 1.1 Hz, 1H), 5.08 (t, J = 1.1 Hz, 2H).

[0060] Generally, the salts are prepared in a manner known per se, by reacting the free acid or base form of the compounds with the appropriate stoichiometric amount of an inorganic or organic base or acid in water or in an organic solvent, or in a mixture of both. Generally, non-aqueous media, such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile, are preferred.

[0061] The preparation method of the AKR1C3-targeted small-molecule conjugate drug according to the present application is selected from any one of the following:

[0062] Method I:

[0063] Specifically, methyl salicylate derivative 1 and 4-chloromethyl-3,5-dimethylisoxazole are used as raw materials, DMF is used as a solvent, potassium carbonate is used as a base, and heating is carried out at 65°C to obtain intermediate 3. Methanol is used as a solvent, a small amount of tetrahydrofuran is used as a cosolvent, lithium hydroxide solution is added dropwise to hydrolyze the ester, and then acid is added to adjust the pH to 1-2 to obtain carboxylic acid intermediate 4. DMF is used as a solvent, HATU is used as a condensing agent, and DIPEA is used as a base. Amide condensation is carried out with intermediate 5 to obtain intermediate 6. Methanol is used as a solvent, a small amount of tetrahydrofuran is used as a cosolvent, lithium hydroxide solution is added dropwise to hydrolyze the ester, and then acid is added to adjust the pH to 1-2 to obtain carboxylic acid intermediate 7.

[0064] Compound 8 and compound 9 are dissolved in DMF solvent, HATU is a condensing agent, DIPEA is a base, and after amide condensation, intermediate 10 is obtained, and after removing the Boc protecting group in 4N HCl / 1,4-dioxane solution, intermediate 11 is obtained, and after amide condensation with intermediate 12 in DMF solvent, HATU is a condensing agent, DIPEA is a base, and after amide condensation, intermediate 13 is obtained, and after removing the Boc protecting group in 4N HCl / 1,4-dioxane solution, intermediate 14 is obtained;

[0065] Intermediate 7 and intermediate 14 are dissolved in DMF solution, HATU is a condensing agent, DIPEA is a base, and after amide condensation, a compound of formula I is obtained;

[0066] Method two:

[0067] Intermediate 7 obtained by method one and intermediate 11 are subjected to amide condensation in DMF solvent, HATU is a condensing agent, and DIPEA is a base, to obtain a compound of formula I;

[0068] wherein R1, R2, R3, R4, L1, L2, D, and n are as defined above.

[0069] The dosage form of the drug is selected from the group consisting of capsules, powders, tablets, granules, pills, injections, syrups, oral solutions, inhalants, ointments, suppositories, and patches. The carrier can be changed according to the dosage form, administration form, etc. The pharmaceutically acceptable carrier includes excipients, binders, disintegrants, lubricants, flavorings, fragrances, colorants, and sweeteners.

[0070] The small molecule conjugated drug or the pharmaceutical composition thereof provided by the application is applied to the preparation of AKR1C3 inhibitors.

[0071] Preferably, the drug is used for treating cancer, including but not limited to preventing cancer, treating cancer, or reversing tumor drug resistance, and is particularly used for treating the hyperproliferation, cell growth inhibition, or cytotoxicity of cancer.

[0072] Beneficial effects: Compared with the prior art, the application has the following remarkable advantages: the compound provided by the application has a novel structure, exhibits excellent targeting and selectivity to AKR1C3, and can realize intelligent response to the tumor microenvironment. The compound has a dual action mechanism of AKR1C3 enzyme inhibition activity and effective payload directly killing tumor cells, and can exhibit a significant anti-tumor effect under single-drug treatment conditions, and has good safety characteristics. BRIEF DESCRIPTION OF DRAWINGS

[0073] Figure 1 AKR1C3 and cathepsin B (CTSB) high-expression tool cells are screened out for Western blotting experiments. Detailed Implementation

[0074] The technical solution of the present invention will be further described below with reference to the embodiments.

[0075] Example 1: Synthesis of 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-(((S)-1-(((S)-1-((4-(((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxopropyl-2-yl)amino)-3-methyl-1-oxobut-2-yl)carbamoyl)phenyl)benzamide (Compound 1)

[0076] (1) Synthesis of methyl 2-((3,5-dimethylisoxazol-4-yl)methoxy)benzoate (intermediate 3a)

[0077] Methyl salicylate 1a (500 mg, 3.29 mmol) and potassium carbonate (1135 mg, 8.22 mmol) were dissolved together in 10 mL of DMF and heated to 65 °C for 30 min. Then, 4-chloromethyl-3,5-dimethylisoxazole (489 μL, 3.94 mmol) was added dropwise, and the reaction continued for 3 h. The reaction was monitored by TLC (electrolyte PE:EA = 3:1, v / v). The reaction ended when the starting material spot disappeared. Post-treatment: Ice water was added to the reaction solution until a large amount of white solid precipitated. The mixture was filtered, and the filter cake was washed several times with water. The filter cake was collected, dried, and the white solid (intermediate 3a, 773 mg, yield 90%) was obtained. 1 H NMR (300MHz, Chloroform-d): δ7.64(dd,J=7.6,2.0Hz,1H),7.32(t,J=7.8Hz,1H),6.99–6.77(m,2H),4.72(s,2H),3.68(s,3H),2.24(s,3H),2.16(s,3H).

[0078] (2) Synthesis of 2-((3,5-dimethylisoxazol-4-yl)methoxy)benzoic acid (intermediate 4a)

[0079] Intermediate 3a (500 mg, 1.91 mmol) was dissolved in 9 mL of methanol and 3 mL of tetrahydrofuran as a dissolving agent. After dissolution, 9 mL of 2NLiOH solution was added dropwise with stirring. The reaction was carried out at room temperature for 24 h. The reaction was monitored by TLC (electrolyte PE:EA = 1:5, v / v). The reaction ended when the starting material spot disappeared. Methanol and tetrahydrofuran were removed under vacuum, and the pH was adjusted to 1-2 at low temperature. A white solid precipitated. The filter cake was collected by suction filtration, dried, and a white solid (intermediate 4a, 435 mg, yield 92%) was obtained, which was directly used for the next synthesis.

[0080] (3) Synthesis of methyl 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido]benzoate (Intermediate 6a)

[0081] Intermediate 4a (300 mg, 1.21 mmol) and HATU (692 mg, 1.82 mmol) were dissolved in 6 mL of DMF, DIPEA (528 μL, 3.03 mmol) was added dropwise and reacted for 5 min, then methyl 3-aminobenzoate 5a (183 mg, 1.21 mmol) was added, and reacted at room temperature for 4 h, TLC monitoring (developing agent PE:EA = 3:1, v / v), the raw material point disappeared, and the reaction was completed. 10 mL of water was added, extracted with ethyl acetate, washed with saturated NaCl solution, dried over anhydrous sodium sulfate, and the ethyl acetate was removed under vacuum. After purification by column chromatography (PE:EA = 3:1, v / v), the white solid was obtained by vacuum drying (Intermediate 6a, 295 mg, yield 64%)

[0082] PE:EA = 3:1, v / v), the raw material point disappeared, and the reaction was completed. 10 mL of water was added, extracted with ethyl acetate, washed with saturated NaCl solution, dried over anhydrous sodium sulfate, and the ethyl acetate was removed under vacuum. After purification by column chromatography (PE:EA = 3:1, v / v), the white solid was obtained by vacuum drying (Intermediate 6a, 295 mg, yield 64%) 1 HNMR (300 MHz, DMSO-d6): δ 10.35 (s, 1H), 8.33 (t, J = 2.1 Hz, 1H), 7.80 (d, J = 7.7 Hz, 1H), 7.71-7.65 (m, 1H), 7.65-7.39 (m, 3H), 7.33 (d, J = 8.3 Hz, 1H), 7.11 (t, J = 7.4 Hz, 1H), 5.04 (s, 2H), 3.86 (s, 3H), 2.39 (s, 3H), 2.17 (s, 3H).

[0083] (4) Synthesis of 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido]benzoic acid (Intermediate 7a)

[0084] Intermediate 6a (150 mg, 0.39 mmol) was dissolved in 9 mL of methanol, 3 mL of tetrahydrofuran was added to assist dissolution, and 2N LiOH solution 9 mL was added dropwise under stirring. After dissolution, it was reacted at room temperature for 24 h, TLC monitoring (developing agent PE:EA = 1:5, v / v), the raw material point disappeared, and the reaction was completed. The methanol and tetrahydrofuran were removed under vacuum, the pH was adjusted to 1-2 at low temperature, and a white solid was precipitated. The filter cake was collected by suction filtration and oven dried to obtain a white solid (Intermediate 7a, 129 mg, yield 89%), which was directly used for the next step synthesis.

[0085] (5) Synthesis of tert-butyl ((S)-1-(((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamate (Intermediate 10a)

[0086] Dissolve (tert-butoxycarbonyl)-L-valyl-L-alanine (300 mg, 1.04 mmol) and HATU (593 mg, 1.56 mmol) in 6 ml DMF, add DIPEA (453 μL, 2.60 mmol) dropwise, react for 5 minutes, then add intermediate F1 (245 mg, 1.04 mmol), react at room temperature for 4 h, monitor the reaction by TLC (developing agent PE:EA = 1:1, v / v), the raw material point disappears, the reaction is completed. Add 10 mL water, extract with ethyl acetate, wash with saturated NaCl solution, dry over anhydrous sodium sulfate, remove the ethyl acetate in vacuum. After purification by column chromatography (PE:EA = 1:1, v / v), dry in vacuum to obtain white solid (intermediate 10a, 300 mg, yield 57%) 1 H NMR (400 MHz, DMSO-d6): δ 11.18 (d, J = 5.9 Hz, 1H), 9.91 (d, J = 75.9 Hz, 1H), 8.20 (dd, J = 73.8, 7.3 Hz, 1H), 7.84 (t, J = 5.5 Hz, 1H), 7.56 (dd, J = 15.4, 8.3 Hz, 2H), 7.32 - 7.21 (m, 2H), 6.77 (dd, J = 37.3, 8.5 Hz, 1H), 4.92 (s, 2H), 4.46 (p, J = 7.1 Hz, 1H), 3.92 - 3.77 (m, 1H), 1.95 (dq, J = 19.9, 6.6 Hz, 1H), 1.36 (d, J = 7.3 Hz, 9H), 1.31 (d, J = 7.0 Hz, 3H), 0.86 (q, J = 6.8 Hz, 6H).

[0087] (6) Synthesis of (S)-2-amino-N-((S)-1-((4-((5-fluoro-2,6-dioxo-3,6- dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxopropan-2-yl)-3- methylbutanamide hydrochloride (intermediate 11a)

[0088] Dissolve intermediate 11a (250 mg, 0.49 mmol) in 5 ml DCM, add 4N HCl / 1,4- Dioxane solution 5 ml, react at room temperature for 4 h, dry in vacuum to obtain yellow solid (intermediate 11a), which is directly used in the next step synthesis.

[0089] (7) Synthesis of 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-(((S)-1-(((S)-1-((4- ((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1- oxopropan-2-yl)amino)-3-methyl-1-oxo-2-yl)carbamoyl)phenyl)benzamide (compound 1)

[0090] Intermediate 11a (100 mg, 0.27 mmol) and HATU (156 mg, 0.41 mmol) were dissolved in 6 mL of DMF, DIPEA (119 μL, 0.68 mmol) was added dropwise, and the reaction was allowed to proceed for 5 min, then intermediate 11a (121 mg, 0.27 mmol) was added, and the reaction was allowed to proceed at room temperature for 4 h. TLC monitoring of the reaction (developing agent DCM:MeOH = 10:1, v / v) showed that the starting material disappeared, and the reaction was completed. 10 mL of water was added, and the product was extracted with ethyl acetate, washed with saturated NaCl solution, dried over anhydrous sodium sulfate, and the ethyl acetate was removed under vacuum. After purification by column chromatography (DCM:MeOH = 10:1, v / v), the product was dried under vacuum to obtain a white solid (compound 1, 51 mg, yield 25%) 1 H NMR (300 MHz, DMSO-d6): δ 11.16 (d, J = 6.0 Hz, 1H), 10.26 (s, 1H), 9.90 (d, J = 41.6 Hz, 1H), 8.54 - 8.08 (m, 3H), 7.83 (q, J = 5.7 Hz, 1H), 7.69 (t, J = 6.4 Hz, 1H), 7.54 (dt, J = 18.1, 8.6 Hz, 5H), 7.38 (d, J = 7.8 Hz, 1H), 7.31 - 7.16 (m, 3H), 7.07 (t, J = 7.6 Hz, 1H), 5.00 (s, 2H), 4.86 (s, 2H), 4.41 (q, J = 7.1 Hz, 1H), 4.28 (dt, J = 24.8, 8.1 Hz, 1H), 2.35 (s, 3H), 2.13 (s, 4H), 1.28 (d, J = 7.0 Hz, 3H), 0.99 - 0.86 (m, 6H).

[0091] Example 2: Synthesis of 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-(((S)-1-(((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamoyl)phenyl)-4-(trifluoromethyl)benzamide (Compound 2)

[0092] (1) Synthesis of methyl 2-((3,5-dimethylisoxazol-4-yl)methoxy)-4- (trifluoromethyl)benzoate (Intermediate 3b)

[0093] Referring to the synthesis method of Example 1, methyl salicylate in Example 1 was replaced with methyl 2-hydroxy-4-trifluoromethylbenzoate to obtain a white solid (Intermediate 3b);

[0094] (2) Synthesis of 2-((3,5-dimethylisoxazol-4-yl)methoxy)-4- (trifluoromethyl)benzoic acid (Intermediate 4b)

[0095] Referring to the synthesis method of Example 1, 2-((3,5-dimethylisoxazol-4- yl)methoxy)benzoic acid in Example 1 was replaced with 2-((3,5-dimethylisoxazol-4- yl)methoxy)-4-(trifluoromethyl)benzoic acid to obtain a white solid (Intermediate 4b);

[0096] (3) Synthesis of methyl 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)-4- (trifluoromethyl)benzamido]benzoate (Intermediate 6b)

[0097] Referring to the synthesis method of Example 1, 2-((3,5-dimethylisoxazol-4- yl)methoxy)benzoic acid in Example 1 was replaced with 2-((3,5-dimethylisoxazol-4- yl)methoxy)-4-(trifluoromethyl)benzoic acid to obtain a white solid (Intermediate 4b);

[0098] (4) Synthesis of 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)-4- (trifluoromethyl)benzamido]benzoic acid (Intermediate 7b)

[0099] Referring to the synthesis method of Example 1, 2-((3,5-dimethylisoxazol-4- yl)methoxy)benzoic acid in Example 1 was replaced with 2-((3,5-dimethylisoxazol-4- yl)methoxy)-4-(trifluoromethyl)benzoic acid to obtain a white solid (Intermediate 4b);

[0100] (5) Synthesis of 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-(((S)-1-(((S)-1- ((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1- oxoprop-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamoyl)phenyl)-4- (trifluoromethyl)benzamide (Compound 2)

[0101] Referring to the synthesis method of Example 1, 2-((3,5-dimethylisoxazol-4- yl)methoxy)benzoic acid in Example 1 was replaced with 2-((3,5-dimethylisoxazol-4- yl)methoxy)-4-(trifluoromethyl)benzoic acid to obtain a white solid (Intermediate 4b); 1H NMR (400 MHz, DMSO-d6) δ 11.18 (s, 1H), 10.49 (d, 1H), 9.92 (d, J = 56.0 Hz, 1H), 8.44 (dd, J = 7.8, 1.7 Hz, 1H), 8.26 (dd, J = 7.8, 1.7 Hz, 1H), 8.15 (d, J = 17.6 Hz, 1H), 7.88 (dd, J = 7.4, 5.5 Hz, 1H), 7.81 - 7.69 (m, 2H), 7.66 (s, 1H), 7.60 (d, J = 8.0 Hz, 2H), 7.53 (d, J = 8.2 Hz, 1H), 7.49 - 7.38 (m, 2H), 7.24 (dd, J = 8.4, 4.7 Hz, 2H), 5.17 (s, 2H), 4.89 (s, 2H), 4.50 - 4.39 (m, 1H), 4.31 (dt, J = 32.0, 8.1 Hz, 1H), 2.40 (s, 3H), 2.21 - 2.07 (m, 4H), 1.32 (d, J = 7.0 Hz, 3H), 1.04 - 0.87 (m, 6H).

[0102] 57.5,7.8Hz,1H),8.26(dd,J=39.5,7.8Hz,1H),8.15(d,J=17.6Hz,1H),7.88(dd,J=7.4,5.5Hz,1H),7.81–7.69(m,2H),7.66(s,1H),7.60(d,J=8.0Hz,2H),7.53(d,J=8.2Hz,1H),7.49–7.38(m,2H),7.24(dd,J=8.4,4.7Hz,2H),5.17(s,2H),4.89(s,2H),4.50–4.39(m,1H),4.31(dt,J=32.0,8.1Hz,1H),2.40(s,3H),2.21–2.07(m,4H),1.32(d,J=7.0Hz,3H),1.04–0.87(m,6H).

[0103] Example 3: Synthesis of 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-(((S)-1-(((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1- oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamoyl)phenyl)-4-methylbenzamide (Compound 3)

[0104] (1) Synthesis of methyl 2-[(3,5-dimethylisoxazol-4-yl)methoxy]-4-methylbenzoate (Intermediate 3c)

[0105] According to the synthetic method of Example 1, methyl 2-((3,5-dimethylisoxazol-4- yl)methoxy)benzoate in Example 1 was replaced with methyl 2-[(3,5-dimethylisoxazol-4- yl)methoxy]-4-methylbenzoate to obtain a white solid (Intermediate 3c);

[0106] (2) Synthesis of 2-[(3,5-dimethylisoxazol-4-yl)methoxy]-4-methylbenzoic acid (Intermediate 4c)

[0107] According to the synthetic method of Example 1, methyl 2-((3,5-dimethylisoxazol-4- yl)methoxy)benzoate in Example 1 was replaced with methyl 2-[(3,5-dimethylisoxazol-4- yl)methoxy]-4-methylbenzoate to obtain a white solid (Intermediate 3c);

[0108] (3) Synthesis of methyl 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)-4- methylbenzamido]benzoate (Intermediate 6c)

[0109] Referring to the synthesis method of Example 1, methyl 3-[2-((3,5-dimethylisoxazol-4- yl)methoxy)benzamido]benzoate in Example 1 was replaced with methyl 3-[2-((3,5- dimethylisoxazol-4-yl)methoxy)-4-methylbenzamido]benzoate to obtain a white solid (Intermediate 6c);

[0110] (4) Synthesis of 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-methylbenzamido]benzoic acid (Intermediate 7c)

[0111] Referring to the synthesis method of Example 1, methyl 3-[2-((3,5-dimethylisoxazol-4- yl)methoxy)benzamido]benzoate in Example 1 was replaced with methyl 3-[2-((3,5- dimethylisoxazol-4-yl)methoxy)-4-methylbenzamido]benzoate to obtain a white solid (Intermediate 6c);

[0112] (5) Synthesis of 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-(((S)-1-(((S)-1-((4-((5- fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxopropan-2- yl)amino)-3-methyl-1-oxobutan-2-yl)carbamoyl)phenyl)-4-methylbenzamide (Compound 3)

[0113] Referring to the synthesis method of Example 1, methyl 3-[2-((3,5-dimethylisoxazol-4- yl)methoxy)benzamido]benzoate in Example 1 was replaced with methyl 3-[2-((3,5- dimethylisoxazol-4-yl)methoxy)-4-methylbenzamido]benzoate to obtain a white solid (Intermediate 6c); 1H NMR (400 MHz, DMSO-d6) δ 10.18 (d, J = 3.1 Hz, 1H), 9.92 (d, J = 54.8 Hz, 1H), 8.53 (d, J = 7.5 Hz, 1H), 8.34 (t, J = 8.3 Hz, 1H), 8.17 (d, J = 21.3 Hz, 1H), 7.86 (t, J = 5.8 Hz, 1H), 7.72 - 7.49 (m, 5H), 7.40 (t, J = 7.9 Hz, 1H), 7.24 (dd, J = 8.3, 5.7 Hz, 2H), 7.18 (s, 1H), 6.94 (d, J = 7.8 Hz, 1H), 5.07 (s, 2H), 4.89 (s, 2H), 4.52 - 4.40 (m, 1H), 4.32 (dt, J = 32.8, 8.0 Hz, 1H), 2.44 - 2.35 (m, 6H), 2.21 - 2.08 (m, 4H), 1.32 (dd, J = 7.1, 2.2 Hz, 3H), 1.03 - 0.84 (m, 6H).

[0114] Example 4: Synthesis of 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-(((S)-1-(((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1- oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamoyl)phenyl)-4-methoxybenzamide (Compound 4)

[0115] (1) Synthesis of methyl 2-[(3,5-dimethylisoxazol-4-yl)methoxy]-4-methoxybenzoate (Intermediate 3d)

[0116] Referring to the synthesis method of Example 1, methyl salicylate in Example 1 was replaced with methyl 4-methoxy salicylate to obtain a white solid (Intermediate 3d);

[0117] (2) Synthesis of 2-[(3,5-dimethylisoxazol-4-yl)methoxy]-4-methoxybenzoic acid (Intermediate 4d)

[0118] Referring to the synthesis method of Example 1, methyl 2-((3,5-dimethylisoxazol-4-yl)methoxy)benzoate in Example 1 was replaced with methyl 2-[(3,5-dimethylisoxazol-4-yl)methoxy]-4-methoxybenzoate to obtain a white solid (Intermediate 4d);

[0119] (3) Synthesis of methyl 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-methoxybenzamido]benzoate (Intermediate 6d)

[0120] Referring to the synthetic method of Example 1, 2-((3,5-dimethylisoxazol-4-yl)methoxy)benzoic acid in Example 1 was replaced with 2-[(3,5-dimethylisoxazol-4-yl)methoxy]-4-methoxybenzoic acid to obtain a white solid (Intermediate 6d);

[0121] (4) Synthesis of 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-methoxybenzamido]benzoic acid (Intermediate 7d)

[0122] Referring to the synthetic method of Example 1, 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido]benzoic acid methyl ester in Example 1 was replaced with 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-methoxybenzamido]benzoic acid methyl ester to obtain a white solid (Intermediate 7d);

[0123] (5) Synthesis of 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-(((S)-1-(((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamoyl)phenyl)-4-methoxybenzamide (Compound 4) Referring to the synthetic method of Example 1, 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido]benzoic acid in Example 1 was replaced with 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-methoxybenzamido]benzoic acid to obtain a white solid (Compound 4) 1 H NMR (400 MHz, DMSO-d6) δ

[0124] 11.33 (s, 1H), 10.19 - 9.94 (m, 2H), 8.65 - 8.24 (m, 2H), 8.17 (dt, J = 25.9, 1.9 Hz, 1H), 7.87 (t, J = 5.1 Hz, 1H), 7.72 - 7.51 (m, 5H), 7.38 (t, J = 7.9, 1.6 Hz, 1H), 7.23 (dd, J = 8.5, 6.7 Hz, 2H), 6.89 (d, J = 2.3 Hz, 1H), 6.70 (dt, J = 8.6, 2.3 Hz, 1H), 5.15 (d, J = 6.3 Hz, 2H), 4.89 (s, 2H), 4.50 - 4.40 (m, 1H), 4.32 (dt, J = 22.5, 8.2 Hz, 1H), 3.86 (s, 3H), 2.42 (s, 3H), 2.25 - 2.09 (m, 4H), 1.32 (dd, J = 7.2, 3.0 Hz, 3H), 1.01 - 0.87 (m, 6H).

[0125] Example 5: Synthesis of 2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-ethoxy-N-(3-(((S)-1-(((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1- oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamoyl)phenyl)benzamide (Compound 5)

[0126] (1) Synthesis of methyl 2-[(3,5-dimethylisoxazol-4-yl)methoxy]-4-ethoxybenzoate (Intermediate 3e)

[0127] Referring to the synthesis method of Example 1, methyl salicylate in Example 1 was replaced with methyl 4-ethoxy salicylate to obtain a white solid (Intermediate 3e);

[0128] (2) Synthesis of 2-[(3,5-dimethylisoxazol-4-yl)methoxy]-4-ethoxybenzoic acid (Intermediate 4d)

[0129] Referring to the synthesis method of Example 1, methyl 2-((3,5-dimethylisoxazol-4-yl)methoxy)benzoate in Example 1 was replaced with methyl 2-[(3,5-dimethylisoxazol-4-yl)methoxy]-4-ethoxybenzoate to obtain a white solid (Intermediate 4e);

[0130] (3) Synthesis of methyl 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-ethoxybenzamido]benzoate (Intermediate 6e)

[0131] The synthesis method of Reference Example 1 was referred to, and 2-((3,5-dimethylisoxazol-4-yl)methoxy)benzoic acid in Example 1 was replaced with 2-[(3,5-dimethylisoxazol-4-yl)methoxy]-4-ethoxybenzoic acid to obtain a white solid (Intermediate 6e);

[0132] (4) Synthesis of 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-ethoxybenzamido]benzoic acid (Intermediate 7e)

[0133] The synthesis method of Reference Example 1 was referred to, and 2-((3,5-dimethylisoxazol-4-yl)methoxy)benzoic acid in Example 1 was replaced with 2-[(3,5-dimethylisoxazol-4-yl)methoxy]-4-ethoxybenzoic acid to obtain a white solid (Intermediate 6e);

[0134] (5) Synthesis of 2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-ethoxy-N-(3-(((S)-1-(((S)-1-((4-

[0135] The synthesis method of Reference Example 1 was referred to, and 2-((3,5-dimethylisoxazol-4-yl)methoxy)benzoic acid in Example 1 was replaced with 2-[(3,5-dimethylisoxazol-4-yl)methoxy]-4-ethoxybenzoic acid to obtain a white solid (Intermediate 6e); 1 H NMR (400 MHz, DMSO-d6) δ

[0136] 11.20 (dd, J = 6.4, 2.7 Hz, 1H), 10.02 (d, J = 19.1 Hz, 1H), 9.86 (s, 1H), 8.53 (d, J = 7.5 Hz, 1H), 8.34 (t, J = 7.7 Hz, 1H), 8.23 - 8.10 (m, 1H), 7.92 - 7.83 (m, 1H), 7.67 (dd, J = 11.4, 8.5 Hz, 1H), 7.63 - 7.48 (m, 4H), 7.39 (t, J = 7.9 Hz, 1H), 7.24 (dd, J = 8.4, 6.2 Hz, 2H), 6.87 (d, J = 2.2 Hz, 1H), 6.70 (dt, J = 8.7, 2.4 Hz, 1H), 5.11 (d, J = 1.8 Hz, 2H), 4.89 (d, J = 2.1 Hz, 2H), 4.51 - 4.40 (m, 1H), 4.31 (dt, J = 32.5, 8.1 Hz, 1H), 4.14 (q, J = 7.0 Hz, 2H), 2.41 (d, J = 2.2 Hz, 3H), 2.21 - 2.07 (m, 4H), 1.37 (dd, J = 6.9 Hz, 6H), 1.01 - 0.88 (m, 6H).

[0137] Example 6: Synthesis of 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-(((S)-1-(((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1- oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamoyl)phenyl)-4-nitrobenzamide (Compound 6)

[0138] (1) Synthesis of methyl 2-[(3,5-dimethylisoxazol-4-yl)methoxy]-4-nitrobenzoate (Intermediate 3f)

[0139] Referring to the synthesis method of Example 1, methyl salicylate in Example 1 was replaced with methyl 4-nitrosalicylate to obtain a white solid (Intermediate 3f);

[0140] (2) Synthesis of 2-[(3,5-dimethylisoxazol-4-yl)methoxy]-4-nitrobenzoic acid (Intermediate 4f)

[0141] Referring to the synthesis method of Example 1, methyl 2-((3,5-dimethylisoxazol-4-yl)methoxy)benzoate in Example 1 was replaced with methyl 2-[(3,5-dimethylisoxazol-4-yl)methoxy]-4-nitrobenzoate to obtain a white solid (Intermediate 4f);

[0142] (3) Synthesis of methyl 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)-4- nitrobenzamido]benzoate (Intermediate 6f)

[0143] Referring to the synthetic method of Example 1, methyl 3-[2-((3,5-dimethylisoxazol-4- yl)methoxy)benzamido]benzoate in Example 1 was replaced with methyl 3-[2-((3,5- dimethylisoxazol-4-yl)methoxy)-4-nitrobenzamido]benzoate to obtain a white solid (Intermediate 6f);

[0144] (4) Synthesis of 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-nitrobenzamido]benzoic acid (Intermediate 7f)

[0145] Referring to the synthetic method of Example 1, methyl 3-[2-((3,5-dimethylisoxazol-4- yl)methoxy)benzamido]benzoate in Example 1 was replaced with methyl 3-[2-((3,5- dimethylisoxazol-4-yl)methoxy)-4-nitrobenzamido]benzoate to obtain a white solid (Intermediate 6f);

[0146] (5) Synthesis of 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-(((S)-1-(((S)-1-((4-((5- fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxopropan-2- yl)amino)-3-methyl-1-oxobutan-2-yl)carbamoyl)phenyl)-4-nitrobenzamide (Compound 6) 1 H NMR (400 MHz, DMSO-d6) δ

[0147] 11.31 (s, 1H), 10.69 (s, 1H), 10.09 (d, J = 56.6 Hz, 1H), 8.54 (dd, J = 54.1, 7.8 Hz, 1H), 8.36 (dd, J = 52.2, 7.9 Hz, 1H), 8.24 - 8.05 (m, 2H), 7.96 (d, J = 8.0 Hz, 1H), 7.89 - 7.74 (m, 3H), 7.65 (dd, J = 24.2, 7.9 Hz, 2H), 7.55 (d, J = 8.2 Hz, 1H), 7.43 (t, J = 7.9 Hz, 1H), 7.23 (dd, J = 8.2, 5.3 Hz, 2H), 5.23 (s, 2H), 4.89 (s, 2H), 4.52 - 4.40 (m, 1H), 4.32 (dt, J = 22.4, 8.1 Hz, 1H), 2.43 (s, 3H), 2.26 - 2.08 (m, 4H), 1.42 - 1.19 (m, 3H), 1.09 - 0.76 (m, 6H).

[0148] Example 7: Synthesis of 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-(((S)-1-(((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1- oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamoyl)phenyl)-4-chlorobenzamide (Compound 7)

[0149] (1) Synthesis of methyl 2-[(3,5-dimethylisoxazol-4-yl)methoxy]-4-chlorobenzoate (Intermediate 3g)

[0150] Referring to the synthesis method of Example 1, methyl salicylate in Example 1 was replaced with methyl 4-chlorosalicylate to obtain a white solid (Intermediate 3g);

[0151] (2) Synthesis of 2-[(3,5-dimethylisoxazol-4-yl)methoxy]-4-chlorobenzoic acid (Intermediate 4g)

[0152] Referring to the synthesis method of Example 1, methyl 2-((3,5-dimethylisoxazol-4-yl)methoxy)benzoate in Example 1 was replaced with methyl 2-[(3,5-dimethylisoxazol-4-yl)methoxy]-4-chlorobenzoate to obtain a white solid (Intermediate 4g);

[0153] (3) Synthesis of methyl 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-chlorobenzamido]benzoate (Intermediate 6g)

[0154] The synthesis method of Reference Example 1 was referred to, and 2-((3,5-dimethylisoxazol-4-yl)methoxy)benzoic acid in Example 1 was replaced with 2-[(3,5-dimethylisoxazol-4-yl)methoxy]-4-chlorobenzoic acid to obtain a white solid (Intermediate 6g);

[0155] (4) Synthesis of 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-chlorobenzoylamino]benzoic acid (Intermediate 7g)

[0156] The synthesis method of Reference Example 1 was referred to, and 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)benzoylamino]benzoic acid methyl ester in Example 1 was replaced with 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-chlorobenzoylamino]benzoic acid methyl ester to obtain a white solid (Intermediate 7g);

[0157] (5) Synthesis of 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-(((S)-1-(((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamoyl)phenyl)-4-chlorobenzamide (Compound 7) The synthesis method of Reference Example 1 was referred to, and 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)benzoylamino]benzoic acid in Example 1 was replaced with 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-chlorobenzoylamino]benzoic acid to obtain a white solid (Compound 7) 1 H NMR (400 MHz, DMSO-d6) δ 11.19 (s, 1H), 10.31 (s, 1H), 9.92 (d, J = 55.9 Hz, 1H), 8.54 - 8.09 (m, 3H), 7.88 (t, J = 5.8 Hz, 1H), 7.76 - 7.68 (m, 1H), 7.66 - 7.56 (m, 3H), 7.55 - 7.36 (m, 3H), 7.29 - 7.22 (m, 2H), 7.18 (d, J = 8.2, 2.1 Hz, 1H), 5.10 (s, 2H), 4.90 (s, 2H), 4.53 - 4.40 (m, 1H), 4.32 (dt, J = 32.6, 8.1 Hz, 1H), 2.40 (s, 3H), 2.19 - 2.09 (m, 4H), 1.32 (dd, 3H), 1.00 - 0.89 (m, 6H).

[0158] Example 8: Synthesis of 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-(((S)-1-(((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamoyl)phenyl)-4-cyanobenzamide (Compound 8)

[0159] (1) Synthesis of methyl 2-[(3,5-dimethylisoxazol-4-yl)methoxy]-4-cyanobenzoate (Intermediate 3h)

[0160] Referring to the synthesis method of Example 1, methyl 2-((3,5-dimethylisoxazol-4-yl)methoxy)benzoate in Example 1 was replaced with methyl 2-[(3,5-dimethylisoxazol-4-yl)methoxy]-4-cyanobenzoate to obtain a white solid (Intermediate 3h);

[0161] (2) Synthesis of 2-[(3,5-dimethylisoxazol-4-yl)methoxy]-4-cyanobenzoic acid (Intermediate 4h)

[0162] Referring to the synthesis method of Example 1, methyl 2-((3,5-dimethylisoxazol-4-yl)methoxy)benzoate in Example 1 was replaced with methyl 2-[(3,5-dimethylisoxazol-4-yl)methoxy]-4-cyanobenzoate to obtain a white solid (Intermediate 3h);

[0163] (3) Synthesis of methyl 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-cyanobenzamido]benzoate (Intermediate 6h)

[0164] Referring to the synthesis method of Example 1, methyl 2-((3,5-dimethylisoxazol-4-yl)methoxy)benzoate in Example 1 was replaced with methyl 2-[(3,5-dimethylisoxazol-4-yl)methoxy]-4-cyanobenzoate to obtain a white solid (Intermediate 3h);

[0165] (4) Synthesis of 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-cyanobenzamido]benzoic acid (Intermediate 7h)

[0166] Referring to the synthesis method of Example 1, methyl 2-((3,5-dimethylisoxazol-4-yl)methoxy)benzoate in Example 1 was replaced with methyl 2-[(3,5-dimethylisoxazol-4-yl)methoxy]-4-cyanobenzoate to obtain a white solid (Intermediate 3h);

[0167] (5) 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-(((S)-1-(((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamoyl)phenyl)-4-cyanobenzamide (Compound 8) was synthesized according to the synthetic procedure of Example 1 by replacing 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido]benzoic acid in Example 1 with 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-cyanobenzamido]benzoic acid to give a white solid (Compound 8) 1 H NMR (600 MHz, DMSO-d6) δ 10.53 (d, J = 7.9 Hz, 1H), 10.24 (s, 1H), 10.12 (s, 1H), 8.32 (d, J = 9.1 Hz, 1H), 7.98 (d, J = 8.6 Hz, 1H), 7.94 - 7.88 (m, 1H), 7.85 (t, J = 2.1 Hz, 1H), 7.67 - 7.60 (m, 2H), 7.60 - 7.52 (m, 6H), 7.49 - 7.37 (m, 2H), 5.38 (d, J = 13.8 Hz, 1H), 5.29 - 5.22 (m, 2H), 4.99 (dt, J = 12.5, 1.0 Hz, 1H), 4.54 - 4.46 (m, 1H), 4.31 (dd, J = 9.1, 6.5 Hz, 1H), 2.34 (d, J = 7.4 Hz, 6H), 2.28 - 2.18 (m, 1H), 1.26 (d, J = 6.8 Hz, 3H), 0.88 (d, J = 6.6 Hz, 3H), 0.83 (d, J = 6.6 Hz, 3H).

[0168] Example 9: Synthesis of 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-(((S)-1-(((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamoyl)phenyl)-5-chlorobenzamide (Compound 9)

[0169] (1) Synthesis of methyl 2-[(3,5-dimethylisoxazol-4-yl)methoxy]-4-chlorobenzoate (Intermediate 3g)

[0170] Methyl 5-chlorosalicylate was used instead of methyl salicylate according to the synthetic procedure of Example 1 to give a white solid (Intermediate 3i);

[0171] (2) Synthesis of 2-[(3,5-dimethylisoxazol-4-yl)methoxy]-5-chlorobenzoic acid (Intermediate 4i)

[0172] Referring to the synthetic method of Example 1, 2-((3,5-dimethylisoxazol-4-yl)methoxy)benzoic acid methyl ester in Example 1 was replaced with 2-[(3,5-dimethylisoxazol-4-yl)methoxy]-5-chlorobenzoic acid methyl ester to obtain a white solid (Intermediate 4i).

[0173] (3) Synthesis of 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)-5-chlorobenzamido]benzoic acid methyl ester (Intermediate 6i)

[0174] Referring to the synthetic method of Example 1, 2-((3,5-dimethylisoxazol-4-yl)methoxy)benzoic acid in Example 1 was replaced with 2-[(3,5-dimethylisoxazol-4-yl)methoxy]-5-chlorobenzoic acid to obtain a white solid (Intermediate 6i).

[0175] (4) Synthesis of 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)-5-chlorobenzamido]benzoic acid (Intermediate 7i)

[0176] Referring to the synthetic method of Example 1, 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido]benzoic acid methyl ester in Example 1 was replaced with 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)-5-chlorobenzamido]benzoic acid methyl ester to obtain a white solid (Intermediate 7i).

[0177] (5) Synthesis of 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-(((S)-1-(((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamoyl)phenyl)-5-chlorobenzamide (Compound 9) Referring to the synthetic method of Example 1, 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido]benzoic acid in Example 1 was replaced with 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)-5-chlorobenzamido]benzoic acid to obtain a white solid (Compound 9) 1H NMR (400 MHz, DMSO-d6) δ 11.20 (s, 1H), 10.40 (d, J = 2.2 Hz, 1H), 9.94 (d, J = 51.4 Hz, 1H), 8.57 - 8.19 (m, 2H), 8.14 (dt, J = 13.8, 1.9 Hz, 1H), 7.93 - 7.83 (m, 1H), 7.75 (d, J = 8.2, 2.6 Hz, 1H), 7.59 (dd, J = 8.2, 6.3 Hz, 4H), 7.53 (d, J = 8.3 Hz, 1H), 7.42 (t, J = 7.9 Hz, 1H), 7.38 - 7.31 (m, 1H), 7.24 (dd, J = 8.6, 3.8 Hz, 2H), 5.05 (s, 2H), 4.90 (s, 2H), 4.52 - 4.40 (m, 1H), 4.32 (dt, J = 28.0, 8.1 Hz, 1H), 2.39 (d, J = 2.3 Hz, 3H), 2.20 - 2.09 (m, 4H), 1.32 (dd, J = 7.1, 2.5 Hz, 3H), 1.04 - 0.89 (m, 6H).

[0178] Example 10: Synthesis of 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-(((S)-1-(((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1- oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamoyl)phenyl)-5-nitrobenzamide (Compound 10)

[0179] (1) Synthesis of methyl 2-[(3,5-dimethylisoxazol-4-yl)methoxy]-5-nitrobenzoate (Intermediate 3j)

[0180] Referring to the synthesis method of Example 1, methyl salicylate in Example 1 was replaced with methyl 5-nitrosalicylate to obtain a white solid (Intermediate 3j);

[0181] (2) Synthesis of 2-[(3,5-dimethylisoxazol-4-yl)methoxy]-5-nitrobenzoic acid (Intermediate 4j)

[0182] Referring to the synthesis method of Example 1, methyl 2-((3,5-dimethylisoxazol-4-yl)methoxy)benzoate in Example 1 was replaced with methyl 2-[(3,5-dimethylisoxazol-4-yl)methoxy]-5-nitrobenzoate to obtain a white solid (Intermediate 4j);

[0183] (3) Synthesis of methyl 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)-5-nitrobenzamido]benzoate (Intermediate 6j)

[0184] Referring to the synthetic method of Example 1, 2-((3,5-dimethylisoxazol-4-yl)methoxy)benzoic acid in Example 1 was replaced with 2-[(3,5-dimethylisoxazol-4-yl)methoxy]-5-nitrobenzoic acid to obtain a white solid (Intermediate 6j);

[0185] (4) Synthesis of 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)-5-nitrobenzamido]benzoic acid (Intermediate 7j)

[0186] Referring to the synthetic method of Example 1, 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido]benzoic acid methyl ester in Example 1 was replaced with 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)-5-nitrobenzamido]benzoic acid methyl ester to obtain a white solid (Intermediate 7j);

[0187] (5) Synthesis of 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-(((S)-1-(((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamoyl)phenyl)-5-nitrobenzamide (Compound 10) Referring to the synthetic method of Example 1, 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido]benzoic acid in Example 1 was replaced with 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)-5-nitrobenzamido]benzoic acid to obtain a white solid (Compound 10) 1 H NMR (600 MHz, DMSO-d6) δ 10.53 (d, J = 7.9 Hz, 1H), 10.34 (s, 1H), 10.11 (s, 1H), 8.49 (d, J = 2.0 Hz, 1H), 8.36 - 8.29 (m, 2H), 7.97 (t, J = 1.9 Hz, 1H), 7.91 - 7.86 (m, 1H), 7.82 - 7.76 (m, 1H), 7.58 - 7.42 (m, 7H), 7.20 (d, J = 8.4 Hz, 1H), 5.35 (d, J = 13.8 Hz, 1H), 5.28 - 5.22 (m, 2H), 4.99 (dt, J = 12.5, 1.0 Hz, 1H), 4.39 - 4.30 (m, 2H), 2.44 (s, 3H), 2.28 - 2.19 (m, 4H), 1.27 (d, J = 6.8 Hz, 3H), 0.88 (d, J = 6.6 Hz, 3H), 0.83 (d, J = 6.6 Hz, 3H).

[0188] Example 11: Synthesis of 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-(((S)-1-(((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamoyl)phenyl)-5-methoxybenzamide (Compound 11)

[0189] (1) Synthesis of methyl 2-[(3,5-dimethylisoxazol-4-yl)methoxy]-5-methoxybenzoate (Intermediate 3k)

[0190] Referring to the synthesis method of Example 1, methyl salicylate in Example 1 was replaced with methyl 5-methoxysalicylate to obtain a white solid (Intermediate 3k);

[0191] (2) Synthesis of 2-[(3,5-dimethylisoxazol-4-yl)methoxy]-5-methoxybenzoic acid (Intermediate 4k)

[0192] Referring to the synthesis method of Example 1, methyl 2-((3,5-dimethylisoxazol-4-yl)methoxy)benzoate in Example 1 was replaced with methyl 2-[(3,5-dimethylisoxazol-4-yl)methoxy]-5-methoxybenzoate to obtain a white solid (Intermediate 4k);

[0193] (3) Synthesis of methyl 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)-5-methoxybenzamido]benzoate (Intermediate 6k)

[0194] Referring to the synthesis method of Example 1, 2-((3,5-dimethylisoxazol-4-yl)methoxy)benzoic acid in Example 1 was replaced with 2-[(3,5-dimethylisoxazol-4-yl)methoxy]-5-methoxybenzoic acid to obtain a white solid (Intermediate 6k);

[0195] (4) Synthesis of 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)-5-methoxybenzamido]benzoic acid (Intermediate 7k)

[0196] Referring to the synthesis method of Example 1, methyl 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido]benzoate in Example 1 was replaced with methyl 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)-5-methoxybenzamido]benzoate to obtain a white solid (Intermediate 7k);

[0197] (5) 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-(((S)-1-(((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamoyl)phenyl)-5-methoxybenzamide (Compound 11) was synthesized according to the synthetic procedure of Example 1 by replacing 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido]benzoic acid in Example 1 with 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)-5-methoxybenzamido]benzoic acid to give a white solid (Compound 11) 1 H NMR (300 MHz, DMSO-d6) δ

[0198] 11.29 (s, 1H), 10.33 (s, 1H), 10.06 (d, J = 35.7 Hz, 1H), 8.58 (d, J = 7.5 Hz, 1H), 8.40 (d, J = 7.8 Hz, 1H), 8.20 (t, J = 13.1 Hz, 1H), 7.87 (d, J = 6.4 Hz, 1H), 7.77 (d, J = 8.1 Hz, 1H), 7.69 - 7.51 (m, 3H), 7.42 (t, J = 7.8 Hz, 1H), 7.34 - 7.20 (m, 3H), 7.20 - 7.05 (m, 2H), 5.01 (s, 2H), 4.90 (s, 2H), 4.46 (s, 1H), 4.42 - 4.25 (m, 1H), 3.79 (d, J = 1.7 Hz, 3H), 2.36 (s, 3H), 2.26 - 2.09 (m, 4H), 1.33 (d, J = 7.2 Hz, 3H), 1.05 - 0.90 (m, 6H).

[0199] Example 12: Synthesis of 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-(((S)-1-(((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamoyl)phenyl)-5-methylbenzamide (Compound 12)

[0200] (1) Synthesis of methyl 2-[(3,5-dimethylisoxazol-4-yl)methoxy]-5-methylbenzoate (Intermediate 31)

[0201] Methyl 5-methylsalicylate was used instead of methyl salicylate according to the synthetic procedure of Example 1 to give a white solid (Intermediate 31);

[0202] (2) Synthesis of 2-[(3,5-dimethylisoxazol-4-yl)methoxy]-5-methylbenzoic acid (Intermediate 4l)

[0203] Referring to the synthetic method of Example 1, 2-((3,5-dimethylisoxazol-4-yl)methoxy)benzoic acid methyl ester in Example 1 was replaced with 2-[(3,5-dimethylisoxazol-4-yl)methoxy]-5-methylbenzoic acid methyl ester to obtain a white solid (Intermediate 4l);

[0204] (3) Synthesis of 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)-5-methylbenzamido]benzoic acid methyl ester (Intermediate 6l)

[0205] Referring to the synthetic method of Example 1, 2-((3,5-dimethylisoxazol-4-yl)methoxy)benzoic acid in Example 1 was replaced with 2-[(3,5-dimethylisoxazol-4-yl)methoxy]-5-methylbenzoic acid to obtain a white solid (Intermediate 6l);

[0206] (4) Synthesis of 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)-5-methylbenzamido]benzoic acid (Intermediate 7l)

[0207] Referring to the synthetic method of Example 1, 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido]benzoic acid methyl ester in Example 1 was replaced with 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)-5-methylbenzamido]benzoic acid methyl ester to obtain a white solid (Intermediate 7l);

[0208] (5) Synthesis of 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-(((S)-1-(((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamoyl)phenyl)-5-methylbenzamide (Compound 12) Referring to the synthetic method of Example 1, 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido]benzoic acid in Example 1 was replaced with 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)-5-methylbenzamido]benzoic acid to obtain a white solid (Compound 12) 1H NMR (600 MHz, DMSO-d6) δ 10.53 (d, J = 7.9 Hz, 1H), 10.11 (s, 1H), 9.13 (s, 1H), 8.32 (d, J = 9.1 Hz, 1H), 8.05 (t, J = 1.9 Hz, 1H), 7.94 - 7.88 (m, 1H), 7.87 - 7.82 (m, 1H), 7.82 - 7.77 (m, 1H), 7.58 - 7.50 (m, 3H), 7.49 - 7.35 (m, 4H), 7.34 - 7.28 (m, 1H), 7.12 (d, J = 8.4 Hz, 1H), 5.43 (d, J = 13.8 Hz, 1H), 5.31 (d, J = 13.7 Hz, 1H), 5.25 (dt, J = 12.5, 1.0 Hz, 1H), 4.99 (dt, J = 12.5, 1.0 Hz, 1H), 4.38 - 4.29 (m, 1H), 4.00 (dd, J = 9.1, 6.5 Hz, 1H), 2.45 (s, 3H), 2.29 - 2.22 (m, 6H), 2.00 - 1.90 (m, 1H), 1.23 (d, J = 6.9 Hz, 3H), 0.88 (d, J = 6.6 Hz, 3H), 0.83 (d, J = 6.6 Hz, 3H).

[0209] Example 13: 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-N-((S)-1-

[0210] Synthesis of ((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1- oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)-5-methoxybenzamide (Compound 13)

[0211] (1) Synthesis of methyl 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-5- methoxybenzoate (Intermediate 6m)

[0212] Referring to the synthesis method of Example 1, methyl 3-aminobenzoate in Example 1 was replaced with methyl 3-amino-5-methoxybenzoate to obtain a white solid (Intermediate 6m);

[0213] (2) Synthesis of 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-5- methoxybenzoic acid (Intermediate 7m)

[0214] Referring to the synthetic method of Example 1, 3-[2-((3,5-dimethylisoxazol-4- yl)methoxy)benzamido]benzoic acid methyl ester in Example 1 was replaced with 3-(2- ((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-5-methoxybenzoic acid methyl ester to obtain a white solid (Intermediate 7m);

[0215] (3) Synthesis of 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-N-((S)-1- (((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1- oxoprop-2-yl)amino)-3-methyl-1-oxobutan-2-yl)-5-methoxybenzamide (Compound 13) Referring to the synthetic method of Example 1, 3-[2-((3,5-dimethylisoxazol-4- yl)methoxy)benzamido]benzoic acid in Example 1 was replaced with 3-(2-((3,5- dimethylisoxazol-4-yl)methoxy)benzamido)-5-methoxybenzoic acid to obtain a white solid (Compound 13) 1 H NMR (300 MHz, DMSO-d6) δ 10.16 (d, J = 2.6 Hz, 1H), 9.80 (d, J = 44.6 Hz, 1H), 8.48 - 8.02 (m, 2H), 7.77 (t, J = 4.9 Hz, 1H), 7.64 (d, J = 12.7 Hz, 1H), 7.57 - 7.36 (m, 4H), 7.32 - 7.19 (m, 2H), 7.14 (dd, J = 8.5, 3.2 Hz, 2H), 7.02 (d, J = 7.6 Hz, 2H), 4.94 (s, 2H), 4.79 (s, 2H), 4.43 - 4.29 (m, 1H), 4.20 (dt, J = 28.3, 8.1 Hz, 1H), 3.67 (d, J = 4.7 Hz, 3H), 2.29 (s, 3H), 2.14 - 1.95 (m, 4H), 1.21 (d, J = 7.1 Hz, 3H), 0.92 - 0.77 (m, 6H).

[0216] Example 14: Synthesis of 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-N- ((S)-1-(((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)- 1-oxoprop-2-yl)amino)-3-methyl-1-oxobutan-2-yl)-5-methylbenzamide (Compound 14)

[0217] (1) Synthesis of 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-5- methylbenzoic acid methyl ester (Intermediate 6n)

[0218] Referring to the synthetic method of Example 1, 3-amino benzoic acid methyl ester in Example 1 was replaced with 3-amino-5-methylbenzoic acid methyl ester to obtain a white solid (Intermediate 6n);

[0219] (2) Synthesis of 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-5- methylbenzoic acid (Intermediate 7n)

[0220] Referring to the synthetic method of Example 1, 3-[2-((3,5-dimethylisoxazol-4- yl)methoxy)benzamido]benzoic acid methyl ester in Example 1 was replaced with 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-5-methylbenzoic acid methyl ester to obtain a white solid (Intermediate 7n);

[0221] (3) Synthesis of 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-N-((S)-1- (((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1- oxoprop-2-yl)amino)-3-methyl-1-oxobutan-2-yl)-5-methylbenzamide (Compound 14)

[0222] Referring to the synthetic method of Example 1, 3-[2-((3,5-dimethylisoxazol-4- yl)methoxy)benzamido]benzoic acid in Example 1 was replaced with 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-5-methylbenzoic acid to obtain a white solid (Compound 14) 1H NMR (400 MHz, DMSO-d6) δ 11.37 (s, 1H), 10.34 (d, J = 2.8 Hz, 1H), 10.12 (d, J = 62.6 Hz, 1H), 8.72 - 8.25 (m, 2H), 8.07 (d, J = 17.9 Hz, 1H), 7.97 (t, J = 5.8 Hz, 1H), 7.79 - 7.58 (m, 5H), 7.54 (d, J = 7.0 Hz, 1H), 7.43 (d, J = 8.3 Hz, 1H), 7.34 (dd, J = 8.3, 5.6 Hz, 2H), 7.28 - 7.13 (m, 1H), 5.17 (d, J = 2.8 Hz, 2H), 5.00 (s, 2H), 4.60 - 4.49 (m, 1H), 4.42 (dt, J = 31.1, 8.1 Hz, 1H), 2.62 (s, 3H), 2.43 (d, J = 3.2 Hz, 3H), 2.35 - 2.17 (m, 4H), 1.43 (d, J = 7.1 Hz, 3H), 1.12 - 0.94 (m, 6H).

[0223] Example 15: 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-N-((S)-1-

[0224] Synthesis of ((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1- oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)-5-hydroxybenzamide (Compound 15)

[0225] (1) Synthesis of methyl 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-5- hydroxybenzoate (Intermediate 6o)

[0226] Referring to the synthetic method of Example 1, methyl 3-aminobenzoate in Example 1 was replaced with methyl 3-amino-5-hydroxybenzoate to obtain a white solid (Intermediate 6o);

[0227] (2) Synthesis of 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-5- hydroxybenzoic acid (Intermediate 7o)

[0228] Referring to the synthetic method of Example 1, methyl 3-[2-((3,5-dimethylisoxazol-4- yl)methoxy)benzamido]benzoate in Example 1 was replaced with methyl 3-(2-((3,5- dimethylisoxazol-4-yl)methoxy)benzamido)-5-hydroxybenzoate to obtain a white solid (Intermediate 7o);

[0229] (3) Synthesis of 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-N-((S)-1- (((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1- oxoprop-2-yl)amino)-3-methyl-1-oxobutan-2-yl)-5-hydroxybenzamide (Compound 15)

[0230] Referring to the synthesis method of Example 1, 3-[2-((3,5-dimethylisoxazol-4- yl)methoxy)benzamido]benzoic acid in Example 1 was replaced with 3-(2-((3,5- dimethylisoxazol-4-yl)methoxy)benzamido)-5-hydroxybenzoic acid to obtain a white solid (Compound 15) 1 H NMR (400 MHz, DMSO-d6) δ

[0231] 10.53 (d, J = 7.9 Hz, 1H), 10.12 (s, 1H), 9.78 (s, 1H), 9.26 (s, 1H), 8.53 (d, J = 9.1 Hz, 1H), 7.90 (dd, J = 8.0, 1.6 Hz, 1H), 7.56 (dt, J = 8.4, 1.0 Hz, 2H), 7.54 - 7.47 (m, 3H), 7.47 - 7.39 (m, 3H), 7.17 - 7.08 (m, 3H), 7.01 (t, J = 2.1 Hz, 1H), 5.40 (d, J = 13.7 Hz, 1H), 5.28 - 5.21 (m, 2H), 4.99 (dt, J = 12.5, 1.0 Hz, 1H), 4.34 - 4.25 (m, 1H), 4.15 (dd, J = 9.1, 6.6 Hz, 1H), 2.42 (s, 3H), 2.23 (s, 3H), 2.15 - 2.06 (m, 1H), 1.25 (d, J = 6.8 Hz, 3H), 0.91 - 0.77 (m, 6H).

[0232] Example 16: Synthesis of 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-N-((S)-1-

[0233] Synthesis of 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-N-((S)-1- (((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1- oxoprop-2-yl)amino)-3-methyl-1-oxobutan-2-yl)-5-nitrobenzamide (Compound 16)

[0234] (1) Synthesis of methyl 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-5- nitrobenzoate (Intermediate 6p)

[0235] Referring to the synthetic method of Example 1, 3-amino benzoic acid methyl ester in Example 1 was replaced with 3-amino-5-nitro benzoic acid methyl ester to obtain a white solid (Intermediate 6p);

[0236] (2) Synthesis of 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-5- nitrobenzoic acid (Intermediate 7p)

[0237] Referring to the synthetic method of Example 1, 3-[2-((3,5-dimethylisoxazol-4- yl)methoxy)benzamido]benzoic acid methyl ester in Example 1 was replaced with 3-(2- ((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-5-nitrobenzoic acid methyl ester to obtain a white solid (Intermediate 7p);

[0238] (3) Synthesis of 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-N-((S)-1- (((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1- oxoprop-2-yl)amino)-3-methyl-1-oxobutan-2-yl)-5-nitrobenzamide (Compound 16)

[0239] Referring to the synthetic method of Example 1, 3-[2-((3,5-dimethylisoxazol-4- yl)methoxy)benzamido]benzoic acid in Example 1 was replaced with 3-(2-((3,5- dimethylisoxazol-4-yl)methoxy)benzamido)-5-nitrobenzoic acid to obtain a white solid (Compound 16) 1 H NMR (600 MHz, DMSO-d6) δ

[0240] 10.53 (d, J = 7.9 Hz, 1H), 10.23 (s, 1H), 10.12 (s, 1H), 8.86 (t, J = 2.2 Hz, 1H), 8.66 (d, J = 9.2 Hz, 1H), 8.30 (t, J = 2.2 Hz, 1H), 8.07 (t, J = 2.2 Hz, 1H), 7.91 - 7.86 (m, 1H), 7.61 - 7.52 (m, 4H), 7.47 (dd, J = 20.1, 7.9 Hz, 1H), 7.42 - 7.36 (m, 2H), 7.17 - 7.10 (m, 2H), 5.40 (d, J = 13.8 Hz, 1H), 5.30 (d, J = 13.7 Hz, 1H), 5.25 (dt, J = 12.5, 1.0 Hz, 1H), 4.99 (dt, J = 12.5, 1.0 Hz, 1H), 4.30 - 4.18 (m, 2H), 2.43 (s, 3H), 2.25 (s, 3H), 2.23 - 2.13 (m, 1H), 1.23 (d, J = 6.8 Hz, 3H), 0.88 (d, J = 6.6 Hz, 3H), 0.83 (d, J = 6.6 Hz, 3H).

[0241] Example 17: Synthesis of 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-N-((S)-1-(((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1- oxoprop-2-yl)amino)-3-methyl-1-oxobutan-2-yl)-4-methoxybenzamide (Compound 17)

[0242] (1) Synthesis of methyl 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-4- methoxybenzoate (Intermediate 6q)

[0243] Referring to the synthetic method of Example 1, methyl 3-aminobenzoate in Example 1 was replaced with methyl 3-amino-4-methoxybenzoate to obtain a white solid (Intermediate 6q);

[0244] (2) Synthesis of 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-4- methoxybenzoic acid (Intermediate 7q)

[0245] Referring to the synthetic method of Example 1, methyl 3-[2-((3,5-dimethylisoxazol-4- yl)methoxy)benzamido]benzoate in Example 1 was replaced with methyl 3-(2-((3,5- dimethylisoxazol-4-yl)methoxy)benzamido)-4-methoxybenzoate to obtain a white solid (Intermediate 7q);

[0246] (3) Synthesis of 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-N-((S)-1-(((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1- oxoprop-2-yl)amino)-3-methyl-1-oxobutan-2-yl)-4-methoxybenzamide (Compound 17)

[0247] Referring to the synthesis method of Example 1, 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido]benzoic acid in Example 1 was replaced with 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-4-methoxybenzoic acid to obtain a white solid (Compound 17) 1 H NMR (400 MHz, DMSO-d6) δ 11.21 (t, J = 5.2 Hz, 1H), 10.23 (d, J = 4.4 Hz, 1H), 9.90 (d, J = 77.3 Hz, 1H), 8.99 (d, J = 25.3 Hz, 1H), 8.65 - 8.19 (m, 2H), 8.14 (t, J = 6.8 Hz, 1H), 7.87 (dt, J = 10.1, 5.4 Hz, 1H), 7.79 - 7.60 (m, 3H), 7.56 (d, J = 8.2 Hz, 1H), 7.42 (d, J = 8.4 Hz, 1H), 7.35 - 7.16 (m, 3H), 7.10 (dd, J = 8.8, 3.5 Hz, 1H), 5.25 (s, 2H), 4.90 (s, 2H), 4.55 - 4.39 (m, 1H), 4.29 (dt, J = 52.8, 8.1 Hz, 1H), 3.58 (s, 3H), 2.46 (s, 3H), 2.24 - 2.03 (m, 4H), 1.34 (d, J = 6.9 Hz, 3H), 1.11 - 0.86 (m, 6H).

[0248] Example 18: Synthesis of 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-N-((S)-1-(((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1- oxoprop-2-yl)amino)-3-methyl-1-oxobutan-2-yl)-4-chlorobenzamide (Compound 18)

[0249] (1) Synthesis of methyl 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-4- chlorobenzoate (Intermediate 6r)

[0250] Referring to the synthetic method of Example 1, 3-amino-4-chlorobenzoic acid methyl ester was used instead of 3-amino benzoic acid methyl ester in Example 1 to obtain a white solid (intermediate 6r);

[0251] (2) Synthesis of 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-4- chlorobenzoic acid (intermediate 7r)

[0252] Referring to the synthetic method of Example 1, 3-(2-((3,5-dimethylisoxazol-4- yl)methoxy)benzamido)-4-chlorobenzoic acid methyl ester was used instead of 3-[2- ((3,5-dimethylisoxazol-4-yl)methoxy)benzamido]benzoic acid methyl ester in Example 1 to obtain a white solid (intermediate 7r);

[0253] (3) Synthesis of 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-N-((S)-1- (((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1- oxoprop-2-yl)amino)-3-methyl-1-oxobutan-2-yl)-4-chlorobenzamide (Compound 18)

[0254] Referring to the synthetic method of Example 1, 3-(2-((3,5-dimethylisoxazol-4- yl)methoxy)benzamido)-4-chlorobenzoic acid was used instead of 3-[2-((3,5- dimethylisoxazol-4-yl)methoxy)benzamido]benzoic acid in Example 1 to obtain a white solid (Compound 18) 1H NMR (600 MHz, DMSO-d6) δ 10.53 (d, J = 7.9 Hz, 1H), 10.12 (s, 1H), 9.96 (s, 1H), 8.27 (d, J = 9.1 Hz, 1H), 8.06 (d, J = 1.9 Hz, 1H), 7.88 (dd, J = 7.9, 1.6 Hz, 1H), 7.69 (dd, J = 8.4, 2.0 Hz, 1H), 7.61 (d, J = 8.4 Hz, 1H), 7.59 - 7.48 (m, 4H), 7.48 - 7.36 (m, 3H), 7.20 - 7.12 (m, 2H), 5.42 (d, J = 13.8 Hz, 1H), 5.32 (d, J = 13.7 Hz, 1H), 5.25 (dt, J = 12.5, 1.0 Hz, 1H), 4.99 (dt, J = 12.5, 1.0 Hz, 1H), 4.48 - 4.40 (m, 1H), 4.22 (dd, J = 9.1, 6.6 Hz, 1H), 2.45 (s, 3H), 2.28 - 2.19 (m, 4H), 1.22 (d, J = 6.8 Hz, 3H), 0.88 (d, J = 6.6 Hz, 3H), 0.83 (d, J = 6.6 Hz, 3H).

[0255] Example 19: 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-N-((S)-1-

[0256] Synthesis of ((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)- yl)methyl)phenyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)-4- methylbenzamide (Compound 19)

[0257] (1) Synthesis of methyl 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-4- methylbenzoate (Intermediate 6s)

[0258] Referring to the synthesis method of Example 1, methyl 3-aminobenzoate in Example 1 was replaced with methyl 3-amino-4-methylbenzoate to obtain a white solid (Intermediate 6s);

[0259] (2) Synthesis of 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-4- methylbenzoic acid (Intermediate 7s)

[0260] Referring to the synthetic method of Example 1, 3-[2-((3,5-dimethylisoxazol-4- yl)methoxy)benzamido]benzoic acid methyl ester in Example 1 was replaced with 3-(2- ((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-4-methylbenzoic acid methyl ester to obtain a white solid (Intermediate 7s);

[0261] (3) Synthesis of 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-N-((S)-1- (((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1- oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)-4-methylbenzamide (Compound 19)

[0262] Referring to the synthetic method of Example 1, 3-[2-((3,5-dimethylisoxazol-4- yl)methoxy)benzamido]benzoic acid methyl ester in Example 1 was replaced with 3-(2- ((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-4-methylbenzoic acid methyl ester to obtain a white solid (Intermediate 7s); 1 H NMR (600 MHz, DMSO-d6) δ

[0263] 10.53 (d, J = 7.9 Hz, 1H), 10.12 (s, 1H), 9.98 (s, 1H), 8.44 (d, J = 9.1 Hz, 1H), 8.22 (d, J = 2.0 Hz, 1H), 7.89 (dd, J = 8.1, 1.7 Hz, 1H), 7.76 (dd, J = 8.4, 1.8 Hz, 1H), 7.62 - 7.57 (m, 2H), 7.57 - 7.41 (m, 5H), 7.38 - 7.32 (m, 1H), 7.17 - 7.10 (m, 2H), 5.43 (d, J = 13.8 Hz, 1H), 5.33 (d, J = 13.8 Hz, 1H), 5.25 (dt, J = 12.5, 1.0 Hz, 1H), 4.99 (dt, J = 12.5, 1.0 Hz, 1H), 4.48 - 4.39 (m, 1H), 4.24 (dd, J = 9.1, 6.6 Hz, 1H), 2.40 (s, 3H), 2.35 (s, 3H), 2.27 (d, J = 1.1 Hz, 3H), 2.14 - 2.04 (m, 1H), 1.23 (d, J = 6.8 Hz, 3H), 0.88 (d, J = 6.6 Hz, 3H), 0.83 (d, J = 6.6 Hz, 3H).

[0264] Example 20: Synthesis of 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-(2-(((S)-1-(((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-2-oxoethyl)phenyl)benzamide (Compound 20)

[0265] ((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-2-oxoethyl)phenyl)benzamide (Compound 20)

[0266] (1) Synthesis of methyl 2-(3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)phenyl)acetate (Intermediate 6t)

[0267] Referring to the synthetic method of Example 1, methyl 3-aminobenzoate in Example 1 was replaced with methyl 3-aminobenzoate to obtain a white solid (Intermediate 6t);

[0268] (2) Synthesis of 2-(3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)phenyl)acetic acid (Intermediate 7t)

[0269] Referring to the synthetic method of Example 1, methyl 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido]benzoate in Example 1 was replaced with methyl 2-(3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)phenyl)acetate to obtain a white solid (Intermediate 7t);

[0270] (3) Synthesis of 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-(2-(((S)-1-(((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-2-oxoethyl)phenyl)benzamide (Compound 20) Referring to the synthetic method of Example 1, 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido]benzoic acid in Example 1 was replaced with 2-(3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)phenyl)acetic acid to obtain a white solid (Compound 20) 1 H NMR (400 MHz, DMSO-d6) δ

[0271] 11.25 (d, J = 6.3 Hz, 1H), 10.13 (s, 1H), 9.94 (d, J = 49.2 Hz, 1H), 8.48 - 8.10 (m, 2H), 7.87 (t, J = 5.9 Hz, 1H), 7.61 - 7.49 (m, 5H), 7.45 - 7.35 (m, 1H), 7.30 (dd, J = 8.4, 2.5 Hz, 1H), 7.21 (q, J = 7.8, 7.1 Hz, 3H), 7.10 (td, J = 7.5, 2.2 Hz, 1H), 7.05 - 6.94 (m, 1H), 5.05 (d, J = 3.0 Hz, 2H), 4.89 (d, J = 2.4 Hz, 2H), 4.47 - 4.33 (m, 1H), 4.16 (dt, 1H), 3.66 - 3.52 (m, 2H), 2.39 (d, J = 3.1 Hz, 3H), 2.18 (d, J = 2.8 Hz, 3H), 2.06 - 1.93 (m, 1H), 1.29 (dd, J = 7.0, 2.2 Hz, 3H), 0.96 - 0.73 (m, 6H).

[0272] Example 21 : 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-((3-(((S)-1-(((S)-1-((4-

[0273] ((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1- oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)aminocarbonyl)phenyl)benzamide (Compound 21)

[0274] Synthesis

[0275] Synthesis of tert-butyl (3-(((S)-1-(((S)-1-((4-((5-fluoro-2,6-dioxo-3,6- dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxopropan-2-yl)amino)-3- methyl-1-oxobutan-2-yl)amino)-3-oxopropyl)carbamate (Intermediate 13u)

[0276] BOC-β-alanine (407 mg, 2.15 mmol) and HATU (1227 mg, 3.23 mmol) were dissolved in 6 ml of DMF, DIPEA (937 μL, 5.38 mmol) was added dropwise and reacted for 5 minutes, then intermediate 11a (951 mg, 2.15 mmol) was added, and the reaction was carried out at room temperature for 4 h, and the reaction was monitored by TLC (developing agent PE:EA = 1:5, v / v), and the raw material point disappeared, and the reaction was completed. 10 mL of water was added, extracted with ethyl acetate, washed with saturated NaCl solution, dried over anhydrous sodium sulfate, and the ethyl acetate was removed under vacuum. After purification by column chromatography (PE:EA = 1:5, v / v), it was dried under vacuum to obtain a white solid (intermediate 13u, 496 mg, yield 41%).

[0277] (2) (S)-2-(3-aminopropanamido)-N-((S)-1-((4-((5-fluoro-2,6-dioxo-3,6- dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxopropan-2-yl)-3- methylbutanamide hydrochloride (intermediate 14u)

[0278] Intermediate 13u (250 mg, 0.44 mmol) was dissolved in 5 ml of DCM, 5 ml of 4N HCl / 1,4-dioxane solution was added, and the reaction was carried out at room temperature for 4 h, and the yellow solid was dried under vacuum and directly used for the next step synthesis.

[0279] (3) Synthesis of 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-((3-(((S)-1-(((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1- oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-3-oxopropyl)carbamoyl)phenyl)benzamide (Compound 21)

[0280] Referring to the synthesis method of Example 1, (S)-2-amino-N-((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1- oxopropan-2-yl)-3-methylbutanamide hydrochloride in Example 1 was replaced with (S)-2-(3-aminopropanamido)-N-((S)-1-((4-((5-fluoro-2,6-dioxo-3,6- dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxopropan-2-yl)-3- methylbutanamide hydrochloride to obtain a white solid (Compound 21) 1H NMR (400 MHz, DMSO-d6) δ 11.19 (s, 1H), 10.28 (s, 1H), 9.87 (d, J = 34.3 Hz, 1H), 8.53 - 8.14 (m, 3H), 8.05 (dd, J = 40.0, 8.1 Hz, 1H), 7.88 (d, J = 5.5 Hz, 1H), 7.66 (d, J = 8.0 Hz, 1H), 7.64 - 7.45 (m, 5H), 7.44 - 7.29 (m, 2H), 7.24 (d, J = 7.9 Hz, 2H), 7.11 (d, J = 7.9 Hz, 1H), 5.05 (s, 2H), 4.90 (s, 2H), 4.41 (dt, J = 15.4, 7.5 Hz, 1H), 4.18 (dt, J = 31.2, 7.5 Hz, 1H), 3.61 - 3.39 (m, 4H), 2.39 (s, 3H), 2.18 (s, 3H), 2.06 - 1.83 (m, 1H), 1.28 (d, J = 11.0, 8.4 Hz, 3H), 0.95 - 0.73 (m, 6H).

[0281] Example 22: Synthesis of 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-((2-(3-(((S)-1-(((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1- oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-3-oxopropoxy)ethyl)carbamoyl)phenyl)benzamide (Compound 22)

[0282] (1) Synthesis of tert-butyl (2-(3-(((S)-1-(((S)-1-((4-((5-fluoro-2,6-dioxo-3,6- dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1- oxobutan-2-yl)amino)-3-oxopropoxy)ethyl)carbamate (Intermediate 13v)

[0283] Referring to the synthesis method of Example 21, BOC-β-alanine in Example 21 is replaced with

[0284] BOC-amino-monoethylene glycol-carboxylic acid, white solid was obtained (Intermediate 13v).

[0285] (2) (S)-2-(3-(2-aminoethoxy)propanamido)-N-((S)-1-((4-((5-fluoro-2,6-dioxo-3,6- dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxopropan-2-yl)-3-methylbutanamide hydrochloride salt (Intermediate 14v)

[0286] Referring to the synthetic procedure of Example 21, tert-butyl (2-(3-(((S)-1-(((S)-1-((4- ((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxopropan- 2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-3-oxopropoxy)ethyl)carbamate was replaced with tert-butyl (3-(((S)-1-(((S)-1-((4-((5-fluoro-2,6-dioxo-3,6- dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1- oxobutan-2-yl)amino)-3-oxopropyl)carbamate to give white solid (Intermediate 14v).

[0287] (3) Synthesis of 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-((2-(3-(((S)-1-(((S)-1-((4- ((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxopropan- 2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-3-oxopropoxy)ethyl)carbamoyl)phenyl) benzamide (Compound 22)

[0288] Referring to the synthetic procedure of Example 1, (S)-2-amino-N-((S)-1-((4-((5-fluoro- 2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxopropan-2-yl)-3- methylbutanamide hydrochloride in Example 1 was replaced with (S)-2-(3-(2- aminoethoxy)propanamido)-N-((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)- yl)methyl)phenyl)amino)-1-oxopropan-2-yl)-3-methylbutanamide hydrochloride to give white solid (Compound 22) 1H NMR (400 MHz, DMSO-d6) δ 10.53 (d, J = 7.9 Hz, 1H), 10.13 (s, 1H), 9.42 (s, 1H), 8.14 (t, J = 4.4 Hz, 1H), 7.95 - 7.87 (m, 3H), 7.82 - 7.74 (m, 2H), 7.58 - 7.48 (m, 6H), 7.48 - 7.42 (m, 1H), 7.31 (dd, J = 20.1, 7.9 Hz, 1H), 7.17 - 7.09 (m, 2H), 5.43 (d, J = 13.8 Hz, 1H), 5.33 (d, J = 13.9 Hz, 1H), 5.25 (dt, J = 12.5, 1.0 Hz, 1H), 4.99 (dt, J = 12.5, 1.0 Hz, 1H), 4.27 - 4.15 (m, 2H), 3.69 (dt, J = 12.2, 5.2 Hz, 1H), 3.63 (dt, J = 11.5, 7.1 Hz, 1H), 3.62 - 3.45 (m, 3H), 3.41 (dt, J = 15.0, 5.2, 4.3 Hz, 1H), 2.48 - 2.33 (m, 5H), 2.31 (s, 3H), 2.11 - 2.00 (m, 1H), 1.16 (d, J = 6.8 Hz, 3H), 0.92 - 0.81 (m, 6H).

[0289] Example 23: Synthesis of 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-((2-(((S)-1-(((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1- oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-2-oxoethyl)carbamoyl)phenyl)benzamide (Compound 23)

[0290] (1) Synthesis of tert-butyl (2-(((S)-1-(((S)-1-((4-((5-fluoro-2,6-dioxo-3,6- dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1- oxobutan-2-yl)amino)-2-oxoethyl)carbamate (Intermediate 13w)

[0291] Referring to the synthesis method of Example 21, BOC-β-alanine in Example 21 is replaced with

[0292] BOC-glycine, white solid was obtained (Intermediate 13w).

[0293] (2) (S)-2-(2-Aminoacetamido)-N-((S)-1-((4-((5-fluoro-2,6-dioxo-3,6- dihydropyrimidin-1 (2H)-yl)methyl)phenyl)amino)-1-oxopropan-2-yl)-3- methylbutanamide hydrochloride (Intermediate 14w)

[0294] Referring to the synthetic procedure of Example 21, tert-butyl (2-(((S)-1-(((S)-1-((4- ((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1 (2H)-yl)methyl)phenyl)amino)-1- oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-2-oxoethyl)carbamate was substituted for tert-butyl (3-(((S)-1-(((S)-1-((4-((5-fluoro-2,6-dioxo-3,6- dihydropyrimidin-1 (2H)-yl)methyl)phenyl)amino)-1-oxopropan-2-yl)amino)-3- methyl-1-oxobutan-2-yl)amino)-3-oxopropyl)carbamate in Example 21 to give (2- (((S)-1-(((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1 (2H)-yl)methyl)phenyl) amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-2-oxoethyl) carbamic acid tert-butyl ester as a white solid (Intermediate 14w).

[0295] (3) 2-((3,5-Dimethylisoxazol-4-yl)methoxy)-N-(3-((2-(((S)-1-(((S)-1-((4-((5-fluoro- 2,6-dioxo-3,6-dihydropyrimidin-1 (2H)-yl)methyl)phenyl)amino)-1-oxopropan-2- yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-2-oxoethyl)carbamoyl)phenyl)benzamide (Compound 23) synthesis

[0296] Referring to the synthetic procedure of Example 1, (S)-2-amino-N-((S)-1-((4-((5- fluoro-2,6-dioxo-3,6-dihydropyrimidin-1 (2H)-yl)methyl)phenyl)amino)-1- oxopropan-2-yl)-3-methylbutanamide hydrochloride in Example 1 was substituted for (S)-2-(2-aminoacetamido)-N-((S)-1-((4-((5-fluoro-2,6-dioxo-3,6- dihydropyrimidin-1 (2H)-yl)methyl)phenyl)amino)-1-oxopropan-2-yl)-3- methylbutanamide hydrochloride to give (S)-2-(2-aminoacetamido)-N-((S)-1-((4- ((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1 (2H)-yl)methyl)phenyl)amino)-1- oxopropan-2-yl)-3-methylbutanamide hydrochloride as a white solid (Compound 23) 1H NMR (600 MHz, DMSO-d6) δ 10.53 (d, J = 7.9 Hz, 1H), 10.11 (s, 1H), 9.38 (s, 1H), 8.61 (t, J = 5.8 Hz, 1H), 8.15 (t, J = 1.9 Hz, 1H), 7.94 - 7.88 (m, 2H), 7.78 (d, J = 9.1 Hz, 1H), 7.64 - 7.48 (m, 6H), 7.48 - 7.42 (m, 3H), 7.19 - 7.10 (m, 2H), 5.50 (d, J = 13.7 Hz, 1H), 5.32 (d, J = 13.8 Hz, 1H), 5.25 (dt, J = 12.5, 1.0 Hz, 1H), 4.99 (dt, J = 12.5, 1.0 Hz, 1H), 4.43 - 4.34 (m, 1H), 3.95 (dd, J = 17.5, 5.8 Hz, 1H), 3.91 - 3.83 (m, 2H), 2.42 (s, 3H), 2.32 (s, 3H), 2.22 - 2.12 (m, 1H), 1.17 (d, J = 6.8 Hz, 3H), 0.89 (d, J = 6.6 Hz, 3H), 0.84 (d, J = 6.6 Hz, 3H).

[0297] Example 24: Synthesis of 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-(((2S,5S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-5-isopropyl-2-methyl-1,4,7-trioxo-10,13-dioxa-3,6-diazapentadec-15-yl)carbamoyl)phenyl)benzamide (Compound 24)

[0298] (1) Synthesis of tert-butyl ((2S,5S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)- yl)methyl)phenyl)amino)-5-isopropyl-2-methyl-1,4,7-trioxo-10,13-dioxa-3,6-diazapentadec-15- yl)carbamate (Intermediate 13x)

[0299] Referring to the synthesis method of Example 21, BOC-β-alanine in Example 21 is replaced with

[0300] BOC-amino-diethyleneglycol-carboxylic acid, white solid was obtained (Intermediate 13x).

[0301] (2) (S)-2-(3-(2-(2-aminoethoxy)ethoxy)propanamido)-N-((S)-1-((4-((5-fluoro- 2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxopropan-2- yl)-3-methylbutanamide hydrochloride (Intermediate 14x)

[0302] Referring to the synthetic procedure of Example 21, tert-butyl (3-(((S)-1-(((S)-1-((4- ((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1- oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)-3-oxopropyl)carbamate in Example 21 was replaced with tert-butyl ((2S,5S)-1-((4-((5-fluoro-2,6-dioxo-3,6- dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-5-isopropyl-2-methyl-1,4,7- trioxo-10,13-dioxo-3,6-diazapentadec-15-yl)carbamate to give a white solid (Intermediate 14x).

[0303] (3) 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-(((2S,5S)-1-((4-((5-fluoro-2,6- dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-5-isopropyl-2- methyl-1,4,7-trioxo-10,13-dioxo-3,6-diazapentadec-15-yl)aminocarbonyl)phenyl) benzamide (Compound 24) was synthesized

[0304] Referring to the synthetic procedure of Example 1, (S)-2-amino-N-((S)-1-((4-((5-fluoro- 2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxopropan-2-yl)- 3-methylbutanamide hydrochloride in Example 1 was replaced with (S)-2-(3-(2-(2- aminoethoxy)ethoxy)propanamido)-N-((S)-1-((4-((5-fluoro-2,6-dioxo-3,6- dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxopropan-2-yl)-3- methylbutanamide hydrochloride to give a white solid (Compound 24) 1H NMR (600 MHz, DMSO-d6) δ 10.53 (d, J = 7.9 Hz, 1H), 10.13 (s, 1H), 9.42 (s, 1H), 8.29 (t, J = 4.4 Hz, 1H), 7.95 - 7.86 (m, 3H), 7.80 (d, J = 9.1 Hz, 1H), 7.76 - 7.70 (m, 1H), 7.61 - 7.56 (m, 2H), 7.56 - 7.39 (m, 6H), 7.18 - 7.11 (m, 1H), 7.07 (dd, J = 7.9, 1.3 Hz, 1H), 5.41 (d, J = 13.7 Hz, 1H), 5.34 (d, J = 13.8 Hz, 1H), 5.25 (dt, J = 12.5, 1.0 Hz, 1H), 4.89 (dt, J = 12.5, 1.0 Hz, 1H), 4.27 - 4.16 (m, 2H), 3.77 - 3.54 (m, 6H), 3.51 - 3.36 (m, 4H), 2.49 - 2.33 (m, 8H), 2.10 - 1.98 (m, J = 6.6 Hz, 1H), 1.26 (d, J = 6.8 Hz, 3H), 0.88 (d, J = 6.7 Hz, 3H), 0.83 (d, J = 6.7 Hz, 3H).

[0305] Example 25: Synthesis of 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-(((S)-1-(((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamoyl)phenyl)benzamide (Compound 25)

[0306] (1) Synthesis of tert-butyl ((S)-1-(((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamate (Intermediate 10y)

[0307] Referring to the synthesis method of Example 1, (tert-butoxycarbonyl)-L-valyl-L-arginine in Example 1 was replaced with (tert-butoxycarbonyl)-L-valyl-L-citrulline to obtain a white solid (Intermediate 10y).

[0308] (2) Synthesis of (S)-2-((S)-2-amino-3-methylbutanamido)-N-(4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)-5-ureidopentanamide hydrochloride (Intermediate 11y)

[0309] Referring to the synthetic procedure of Example 1, tert-butyl ((S)-1-(((S)-1-((4-((5-fluoro- 2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxo-5-ureidopentan-2- yl)amino)-3-methyl-1-oxobutan-2-yl)carbamate in Example 1 was replaced with tert-butyl ((S)-1-(((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)- 1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamate to give a yellow solid (Intermediate 11y) which was used directly for the next reaction.

[0310] (3) Synthesis of 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-(((S)-1-(((S)-1-((4-((5-fluoro- 2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxo-5-ureidopentan-2- yl)amino)-3-methyl-1-oxobutan-2-yl)carbamoyl)phenyl)benzamide (Compound 25)

[0311] Referring to the synthetic procedure of Example 1, (S)-2-amino-N-((S)-1-((4-((5-fluoro- 2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxopropan-2-yl)-3- methylbutanamide hydrochloride in Example 1 was replaced with (S)-2-((S)-2-amino-3- methylbutanamido)-N-(4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)- 5-ureidopentanamide hydrochloride to give a white solid (Compound 25) 1H NMR (400 MHz, DMSO-d6) δ 10.53 (d, J = 7.9 Hz, 1H), 10.19 (s, 1H), 9.39 (s, 1H), 8.32 (d, J = 9.1 Hz, 1H), 7.94 - 7.88 (m, 2H), 7.85 (t, J = 2.3 Hz, 1H), 7.66 - 7.61 (m, 1H), 7.58 - 7.48 (m, 4H), 7.48 - 7.43 (m, 3H), 7.38 (dd, J = 20.1, 8.1 Hz, 1H), 7.18 - 7.11 (m, 2H), 6.20 (t, J = 4.4 Hz, 1H), 5.82 (s, 2H), 5.52 (d, J = 13.7 Hz, 1H), 5.40 (d, J = 13.8 Hz, 1H), 5.25 (dt, J = 12.5, 1.0 Hz, 1H), 4.99 (dt, J = 12.5, 1.0 Hz, 1H), 4.25 (dt, J = 9.3, 5.8 Hz, 1H), 3.87 (dd, J = 9.1, 6.5 Hz, 1H), 3.26 - 3.17 (m, 1H), 3.15 - 3.06 (m, 1H), 2.42 (s, 3H), 2.35 (s, 3H), 2.27 - 2.16 (m, J = 6.6 Hz, 1H), 1.87 - 1.78 (m, 1H), 1.75 - 1.62 (m, 2H), 1.45 - 1.34 (m, 1H), 0.93 - 0.82 (m, 6H).

[0312] Example 26: 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-N-((S)-1-

[0313] Synthesis of (((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1- oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)-5-methoxybenzamide (Compound 26)

[0314] Referring to the synthesis of Example 13, replace intermediate 11a in Example 13 with intermediate 11y to give a white solid (Compound 26) 1H NMR (600 MHz, DMSO-d6) δ 10.53 (d, J = 7.9 Hz, 1H), 10.21 (s, 1H), 9.64 (s, 1H), 8.40 (d, J = 9.1 Hz, 1H), 7.90 (dd, J = 7.8, 1.4 Hz, 1H), 7.78 - 7.72 (m, 2H), 7.57 - 7.49 (m, 6H), 7.41 (dd, J = 20.0, 8.0 Hz, 1H), 7.20 - 7.11 (m, 3H), 6.20 (t, J = 4.4 Hz, 1H), 5.82 (s, 2H), 5.55 (d, J = 13.7 Hz, 1H), 5.41 (d, J = 13.7 Hz, 1H), 5.25 (dt, J = 12.5, 1.0 Hz, 1H), 4.99 (dt, J = 12.5, 1.0 Hz, 1H), 4.35 (dt, J = 9.3, 5.8 Hz, 1H), 3.91 (dd, J = 9.1, 6.4 Hz, 1H), 3.78 (s, 3H), 3.24 - 3.15 (m, 1H), 3.15 - 3.07 (m, 1H), 2.35 (s, 3H), 2.27 (s, 3H), 2.18 - 2.06 (m, J = 6.6 Hz, 1H), 1.87 - 1.78 (m, 1H), 1.70 - 1.58 (m, 2H), 1.58 - 1.49 (m, 1H), 0.89 (d, J = 6.7 Hz, 3H), 0.84 (d, J = 6.7 Hz, 3H).

[0315] Example 27: Synthesis of (S)-2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-((2-((2-((1-((2-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-2- oxoehtyl)amino)-1-oxo-3-phenylpropan-2-yl)amino)-2-oxoethyl)amino)-2-oxoethyl)carbamoyl)phenyl)benzamide (Compound 27)

[0316] (1) Synthesis of (S)-(2-((2-((1-((2-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)- yl)methyl)phenyl)amino)-2-oxoethyl)amino)-1-oxo-3-phenylpropan-2-yl)amino)-2-oxoethyl)amino)-2- oxoethyl)carbamic acid tert-butyl ester (Intermediate 10z)

[0317] Following the synthetic procedure of Example 1, (tert-butoxycarbonyl)-L-valyl-L-alanine in Example 1 was replaced with (tert-butoxycarbonyl)-glycyl-glycyl-L-phenylalanyl-L-glycine to give (Intermediate 10z) as a white solid.

[0318] (2) Synthesis of (S)-2-(2-(2-aminoacetamido)acetamido)-N-(2-((4-((5-fluoro- 2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-2-oxoethyl)-3- phenylpropanamide hydrochloride (Intermediate 11z)

[0319] Referring to the synthetic procedure of Example 1, tert-butyl ((S)-1-(((S)-1-((4-((5- fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1- oxoprop-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamate in Example 1 was replaced with tert-butyl (S)-(2-((2-((1-((2-((4-((5-fluoro-2,6-dioxo-3,6- dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-2-oxoethyl)amino)-1-oxo-3- phenylprop-2-yl)amino)-2-oxoethyl)amino)-2-oxoethyl)carbamate to give a yellow solid (Intermediate 11z) which was used directly in the next reaction.

[0320] (3) Synthesis of (S)-2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-((2-((2-((1-((2- ((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-2- oxoethyl)amino)-1-oxo-3-phenylprop-2-yl)amino)-2-oxoethyl)amino)-2- oxoethyl)carbamoyl)phenyl)benzamide (Compound 27)

[0321] Referring to the synthetic procedure of Example 1, (S)-2-amino-N-((S)-1-((4-((5- fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1- oxoprop-2-yl)-3-methylbutanamide hydrochloride in Example 1 was replaced with (S)-2-(2-(2-aminoacetamido)acetamido)-N-(2-((4-((5-fluoro-2,6-dioxo-3,6- dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-2-oxoethyl)-3-phenylpropanamide hydrochloride to give a white solid (Compound 27) 1H NMR (400 MHz, DMSO-d6) δ 10.53 (d, J = 7.9 Hz, 1H), 10.46 (s, 1H), 9.38 (s, 1H), 8.60 (t, J = 5.8 Hz, 1H), 8.12 (dt, J = 15.5, 5.8 Hz, 2H), 8.03 (d, J = 9.2 Hz, 1H), 7.94 - 7.87 (m, 2H), 7.73 (t, J = 1.9 Hz, 1H), 7.63 - 7.56 (m, 2H), 7.58 - 7.42 (m, 6H), 7.28 - 7.17 (m, 5H), 7.17 - 7.11 (m, 1H), 7.09 (dd, J = 7.9, 1.3 Hz, 1H), 5.43 (d, J = 13.7 Hz, 1H), 5.34 (d, J = 13.7 Hz, 1H), 5.25 (dt, J = 12.5, 1.0 Hz, 1H), 4.99 (dt, J = 12.5, 1.0 Hz, 1H), 4.67 (dt, J = 9.4, 7.7 Hz, 1H), 4.05 (dd, J = 17.5, 5.8 Hz, 1H), 4.00 - 3.83 (m, 4H), 3.77 (dd, J = 17.5, 5.8 Hz, 1H), 3.08 - 3.00 (m, 1H), 2.90 - 2.82 (m, 1H), 2.41 (s, 3H), 2.22 (s, 3H).

[0322] Example 28: 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-N-((S)-1-

[0323] Synthesis of (((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)-5-methoxybenzamide (Compound 28)

[0324] Referring to the synthesis of Example 13, replace intermediate 11a in Example 13 with intermediate 11z to give a white solid (Compound 28) 1H NMR (600 MHz, DMSO-d6) δ 10.53 (d, J = 7.9 Hz, 1H), 10.43 (s, 1H), 9.62 (s, 1H), 8.62 (t, J = 5.8 Hz, 1H), 8.13 (dt, J = 20.9, 5.8 Hz, 2H), 8.03 (d, J = 9.2 Hz, 1H), 7.90 (dd, J = 7.8, 1.6 Hz, 1H), 7.84 (t, J = 2.1 Hz, 1H), 7.58 - 7.51 (m, 3H), 7.46 (dd, J = 20.1, 7.9 Hz, 1H), 7.32 (dt, J = 8.3, 1.1 Hz, 2H), 7.28 - 7.17 (m, 7H), 7.17 - 7.08 (m, 2H), 5.47 (d, J = 13.7 Hz, 1H), 5.40 (d, J = 13.8 Hz, 1H), 5.12 (dt, J = 12.5, 1.0 Hz, 1H), 4.99 (dt, J = 12.5, 1.1 Hz, 1H), 4.42 (dt, J = 9.2, 7.7 Hz, 1H), 4.09 (dd, J = 17.5, 5.8 Hz, 1H), 3.97 (dd, J = 17.5, 5.8 Hz, 1H), 3.93 - 3.80 (m, 3H), 3.80 - 3.74 (m, 4H), 3.08 - 3.00 (m, 1H), 2.90 - 2.82 (m, 1H), 2.43 (s, 3H), 2.39 (s, 3H).

[0325] Example 29: 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-(((S)-1-(((S)-1-((4-((5-fluoro- 2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxopropan-2-yl)amino)- 3-methyl-1-oxobutan-2-yl)carbamoyl)-5-nitrophenyl)-4-nitrobenzamide (Compound 29)

[0326] Synthesis

[0327] (1) Synthesis of methyl 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-nitrobenzamido)- 5-nitrobenzoate (Intermediate 6rs)

[0328] Referring to Example 6, methyl 3-aminobenzoate in Example 6 was replaced with methyl 3-amino-5-nitrobenzoate to obtain a white solid (Intermediate 6rs);

[0329] (2) Synthesis of 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-nitrobenzamido)-5- nitrobenzoic acid (Intermediate 7rs)

[0330] Reference Example 6, 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-nitrobenzamido]benzoic acid methyl ester in Example 6 was replaced with 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-nitrobenzamido)-5-nitrobenzoic acid methyl ester to obtain a white solid (Intermediate 7rs);

[0331] (3) Synthesis of 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-(((S)-1-(((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamoyl)-5-nitrophenyl)-4-nitrobenzamide (Compound 29) Reference Example 6, 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-nitrobenzamido]benzoic acid in Example 6 was replaced with 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-nitrobenzamido)-5-nitrobenzoic acid to obtain a white solid (Compound 29) 1 H NMR (600 MHz, DMSO-d6) δ

[0332] 11.14 (s, 1H), 10.53 (d, J = 7.9 Hz, 1H), 10.12 (s, 1H), 8.85 (t, J = 2.1 Hz, 1H), 8.66 (d, J = 9.1 Hz, 1H), 8.37 (t, J = 2.1 Hz, 1H), 8.32 (t, J = 2.1 Hz, 1H), 8.19 (d, J = 8.4 Hz, 1H), 8.13 (dd, J = 8.4, 1.8 Hz, 1H), 7.93 (d, J = 1.9 Hz, 1H), 7.63 - 7.51 (m, 5H), 7.43 (dd, J = 20.1, 8.1 Hz, 1H), 5.43 (d, J = 13.8 Hz, 1H), 5.33 (d, J = 13.8 Hz, 1H), 5.25 (dt, J = 12.5, 1.0 Hz, 1H), 4.99 (dt, J = 12.5, 1.0 Hz, 1H), 4.29 - 4.18 (m, 2H), 2.45 (s, 3H), 2.20 - 2.11 (m, 4H), 1.21 (d, J = 6.8 Hz, 3H), 0.89 (d, J = 6.7 Hz, 3H), 0.84 (d, J = 6.7 Hz, 3H).

[0333] Example 30: Synthesis of 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-(((S)-1-(((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1- oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamoyl)-5-methoxyphenyl)-4- nitrobenzamide (Compound 30)

[0334] (1) Synthesis of methyl 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-nitrobenzamido)-5- methoxybenzoate (Intermediate 6rt)

[0335] Referring to Example 6, methyl 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)-4- nitrobenzamido]benzoate in Example 6 was replaced with methyl 3-(2-((3,5-dimethylisoxazol-4- yl)methoxy)-4-nitrobenzamido)-5-methoxybenzoate to obtain a white solid (Intermediate 6rt);

[0336] (2) Synthesis of 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-4-nitrobenzamido)-5- methoxybenzoic acid (Intermediate 7rt)

[0337] Referring to Example 6, methyl 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)-4- nitrobenzamido]benzoate in Example 6 was replaced with methyl 3-(2-((3,5-dimethylisoxazol-4- yl)methoxy)-4-nitrobenzamido)-5-methoxybenzoate to obtain a white solid (Intermediate 6rt);

[0338] (3) Synthesis of 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-(((S)-1-(((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1- oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamoyl)-5-methoxyphenyl)-4- nitrobenzamide (Compound 30)

[0339] Referring to Example 6, methyl 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)-4- nitrobenzamido]benzoate in Example 6 was replaced with methyl 3-(2-((3,5-dimethylisoxazol-4- yl)methoxy)-4-nitrobenzamido)-5-methoxybenzoate to obtain a white solid (Intermediate 6rt); 1H NMR (600 MHz, DMSO-d6) δ 10.57 - 10.51 (m, 2H), 10.11 (s, 1H), 8.40 (d, J = 9.1 Hz, 1H), 8.29 (d, J = 8.4 Hz, 1H), 8.11 (dd, J = 8.4, 2.0 Hz, 1H), 7.92 (d, J = 2.0 Hz, 1H), 7.73 (dd, J = 20.1, 7.9 Hz, 1H), 7.62 - 7.55 (m, 3H), 7.52 (d, J = 9.2 Hz, 1H), 7.44 - 7.38 (m, 3H), 7.08 (t, J = 2.1 Hz, 1H), 5.44 (d, J = 13.8 Hz, 1H), 5.32 (d, J = 13.7 Hz, 1H), 5.12 (dt, J = 12.5, 1.0 Hz, 1H), 4.99 (dt, J = 12.5, 1.1 Hz, 1H), 4.26 - 4.17 (m, 1H), 3.82 (dd, J = 9.1, 6.6 Hz, 1H), 3.79 (s, 3H), 2.41 (s, 3H), 2.16 - 2.07 (m, 1H), 2.02 (s, 3H), 1.18 (d, J = 6.9 Hz, 3H), 0.89 (d, J = 6.7 Hz, 3H), 0.84 (d, J = 6.7 Hz, 3H).

[0340] Example 31: Synthesis of 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-(((S)-1-(((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1- oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamoyl)-5-methoxyphenyl)-4- methoxybenzamide (Compound 31)

[0341] (1) Synthesis of methyl 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-4- methoxybenzamido)-5-methoxybenzoate (Intermediate 6ru)

[0342] Referring to Example 4, methyl 3-aminobenzoate in Example 4 is replaced with methyl 3-amino-5-methoxybenzoate to obtain a white solid (Intermediate 6ru);

[0343] (2) Synthesis of 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)-4- methoxybenzamido)-5-methoxybenzoic acid (Intermediate 7ru)

[0344] Reference Example 4 was repeated, replacing 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)-4- methoxybenzamido]benzoic acid methyl ester in Example 4 with 3-(2-((3,5-dimethylisoxazol-4- yl)methoxy)-4-methoxybenzamido)-5-methoxybenzoic acid methyl ester, to give a white solid (intermediate 7ru);

[0345] (3) Synthesis of 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-(((S)-1-(((S)-1-((4-((5-fluoro- 2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxopropan-2-yl)amino)-3- methyl-1-oxobutan-2-yl)carbamoyl)-5-methoxyphenyl)-4-methoxybenzamide (Compound 31)

[0346] Reference Example 4 was repeated, replacing 3-[2-((3,5-dimethylisoxazol-4-yl)methoxy)-4- methoxybenzamido]benzoic acid methyl ester in Example 4 with 3-(2-((3,5-dimethylisoxazol-4- yl)methoxy)-4-methoxybenzamido)-5-methoxybenzoic acid methyl ester, to give a white solid (intermediate 7ru); 1 H NMR (400 MHz, DMSO-d6) d 11.26 (s, 1H), 10.15 - 9.81 (m, 2H), 8.52 (dd, J = 70.1, 7.7 Hz, 1H), 8.31 (dd, J = 58.7, 7.7 Hz, 1H), 8.05 - 7.77 (m, 2H), 7.76 - 7.63 (m, 2H), 7.54 (d, J = 8.2 Hz, 1H), 7.32 - 7.08 (m, 4H), 6.89 (d, J = 2.3 Hz, 1H), 6.71 (dd, J = 8.7, 2.5 Hz, 1H), 5.14 (d, J = 3.9 Hz, 2H), 4.89 (s, 2H), 4.44 (q, J = 6.9 Hz, 1H), 4.30 (dt, J = 32.2, 8.1 Hz, 1H), 3.86 (s, 3H), 3.77 (d, J = 3.9 Hz, 3H), 2.43 (s, 3H), 2.26 - 2.07 (m, 4H), 1.32 (d, J = 7.1 Hz, 3H), 1.06 - 0.86 (m, 6H).

[0347] Example 32: Synthesis of N-(3-(((S)-l-(((S)-l-((l-((2R,4R,5R)-3,3-difluoro-4- hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-2-oxo-l,2-dihydropyrimidin-4- yl)amino)-l-oxopropan-2-yl)amino)-3-methyl-l-oxobutan-2-yl)carbamoyl)phenyl)-2- ((3,5-dimethylisoxazol-4-yl)methoxy)benzamide (Compound 32)

[0348] (1) Synthesis of tert-butyl (S)-2-amino-N-((S)-l-((l-((2R,4R,5R)-3,3-difluoro-4- hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-2-oxo-l,2-dihydropyrimidin-4- yl)amino)-l-oxopropan-2-yl)-3-methylbutanamide (Intermediate 10rv)

[0349] Referring to the synthetic procedure of Example 1, tert-butyl ((S)-l-(((S)-l-((4-((5- fluoro-2,6-dioxo-3,6-dihydropyrimidin-l(2H)-yl)methyl)phenyl)amino)-l- oxopropan-2-yl)amino)-3-methyl-l-oxobutan-2-yl)carbamate in Example 1 was replaced with tert-butyl ((S)-l-(((S)-l-((l-((2R,4R,5R)-3,3-difluoro-4-hydroxy-5- (hydroxymethyl)tetrahydrofuran-2-yl)-2-oxo-l,2-dihydropyrimidin-4-yl)amino)-l- oxopropan-2-yl)amino)-3-methyl-l-oxobutan-2-yl)carbamate to give a yellow solid (Intermediate 11rv) which was used directly in the next reaction.

[0350] (2) Synthesis of (S)-2-amino-N-((S)-l-((l-((2R,4R,5R)-3,3-difluoro-4-hydroxy-5- (hydroxymethyl)tetrahydrofuran-2-yl)-2-oxo-l,2-dihydropyrimidin-4-yl)amino)-l- oxopropan-2-yl)-3-methylbutanamide hydrochloride (Intermediate 11rv)

[0351] Referring to the synthetic procedure of Example 1, tert-butyl ((S)-l-(((S)-l-((4-((5- fluoro-2,6-dioxo-3,6-dihydropyrimidin-l(2H)-yl)methyl)phenyl)amino)-l- oxopropan-2-yl)amino)-3-methyl-l-oxobutan-2-yl)carbamate in Example 1 was replaced with tert-butyl ((S)-l-(((S)-l-((l-((2R,4R,5R)-3,3-difluoro-4-hydroxy-5- (hydroxymethyl)tetrahydrofuran-2-yl)-2-oxo-l,2-dihydropyrimidin-4-yl)amino)-l- oxopropan-2-yl)amino)-3-methyl-l-oxobutan-2-yl)carbamate to give a yellow solid (Intermediate 11rv) which was used directly in the next reaction.

[0352] (3) Synthesis of N-(3-(((S)-l-(((S)-l-((l-((2R,4R,5R)-3,3-difluoro-4-hydroxy-5- (hydroxymethyl)tetrahydrofuran-2-yl)-2-oxo-l,2-dihydropyrimidin-4-yl)amino)-l- oxopropan-2-yl)amino)-3-methyl-l-oxobutan-2-yl)carbamoyl)phenyl)-2-((3,5- dimethylisoxazol-4-yl)methoxy)benzamide (Compound 32)

[0353] Referring to the synthesis method of Example 1, (S)-2-amino-N-((S)-l-((4-((5-fluoro- 2,6-dioxo-3,6-dihydropyrimidin-l(2H)-yl)methyl)phenyl)amino)-l-oxopropan-2-yl)-3- methylbutanamide hydrochloride in Example 1 was replaced with (S)-2-amino-N-((S)-l- ((l-((2R,4R,5R)-3,3-difluoro-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-2- oxo-l,2-dihydropyrimidin-4-yl)amino)-l-oxopropan-2-yl)-3-methylbutanamide hydrochloride to obtain a white solid (Compound 32) 1 H NMR (600 MHz, DMSO-d6) δ 9.67 (s, 1H), 9.40 (s, 1H), 8.32 (d, J = 9.1 Hz, 1H), 8.11 (t, J = 1.9 Hz, 1H), 7.94 - 7.88 (m, 2H), 7.86 - 7.80 (m, 1H), 7.77 (d, J = 7.6 Hz, 1H), 7.58 (d, J = 9.2 Hz, 1H), 7.53 - 7.48 (m, 1H), 7.45 (t, J = 7.8 Hz, 1H), 7.20 - 7.14 (m, 1H), 7.12 (dd, J = 7.8, 1.3 Hz, 1H), 6.98 (s, OH), 6.94 (s, OH), 6.91 (s, OH), 6.38 (d, J = 7.7 Hz, 1H), 5.66 (d, J = 4.9 Hz, 1H), 5.43 (d, J = 13.7 Hz, 1H), 5.34 (d, J = 13.7 Hz, 1H), 4.76 (t, J = 7.1 Hz, 1H), 4.62 - 4.49 (m, 1H), 4.23 - 4.13 (m, 2H), 4.08 (dt, J = 6.6, 4.8 Hz, 1H), 3.84 - 3.77 (m, 1H), 3.51 (ddd, J = 11.9, 7.1, 4.7 Hz, 1H), 2.46 (s, 3H), 2.30 - 2.20 (m, 4H), 1.29 (d, J = 6.8 Hz, 3H), 0.88 (d, J = 6.6 Hz, 3H), 0.83 (d, J = 6.6 Hz, 3H).

[0354] Example 33: Synthesis of N-((S)-l-(((S)-l-((l-((2R,4R,5R)-3,3-difluoro-4- hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-2-oxo-l,2-dihydropyrimidin-4- yl)amino)-l-oxopropan-2-yl)amino)-3-methyl-l-oxobutan-2-yl)-3-(2-((3,5- dimethylisoxazol-4-yl)methoxy)benzamido)-5-methoxybenzamide (Compound 33)

[0355] Referring to the synthesis of Example 13, intermediate 11a in Example 13 was replaced with intermediate 11rv to give a white solid (Compound 33) 1 H NMR (600 MHz, DMSO-d6) δ 11.02 (d, J = 40.7 Hz, 1H), 10.23 (d, J = 5.6 Hz, 1H), 8.53 - 8.40 (m, 1H), 8.27 (d, J = 7.6 Hz, 1H), 8.17 (dd, J = 24.4, 8.5 Hz, 1H), 7.81 - 7.68 (m, 1H), 7.58 (d, J = 7.1 Hz, 1H), 7.52 (t, J = 7.9 Hz, 1H), 7.47 - 7.36 (m, 1H), 7.32 (dd, J = 8.4, 2.2 Hz, 1H), 7.26 (dd, J = 7.7, 5.2 Hz, 1H), 7.15 (dt, J = 11.4, 1.8 Hz, 1H), 7.11 (t, J = 7.5 Hz, 1H), 6.34 (dd, J = 6.6, 2.4 Hz, 1H), 6.25 - 6.02 (m, 1H), 5.31 (q, J = 5.1 Hz, 1H), 5.05 (d, J = 3.7 Hz, 2H), 4.55 - 4.41 (m, 1H), 4.36 (t, J = 8.4 Hz, 1H), 4.26 - 4.12 (m, 1H), 3.95 - 3.87 (m, 1H), 3.79 (s, 4H), 3.72 - 3.62 (m, 1H), 2.39 (d, J = 2.6 Hz, 3H), 2.22 - 2.09 (m, 4H), 1.32 (t, J = 7.4 Hz, 3H), 1.01 - 0.87 (m, 6H).

[0356] Example 34: Synthesis of N-(3-(2-(((S)-l-(((S)-l-((l-((2R,4R,5R)-3,3-difluoro-4- hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-2-oxo-l,2-dihydropyrimidin-4- yl)amino)-l-oxopropan-2-yl)amino)-3-methyl-l-oxobutan-2-yl)amino)-2- oxoethyl)phenyl)-2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamide (Compound 34)

[0357] Referring to the synthesis of Example 20, replace intermediate 11a in Example 20 with intermediate 11rv to give a white solid (Compound 34) 1 H NMR (600 MHz, DMSO-d6) δ 9.67 (s, 1H), 8.37 (s, 1H), 7.88 (dd, J = 7.8, 1.5 Hz, 1H), 7.77 (d, J = 7.6 Hz, 1H), 7.72 (d, J = 9.1 Hz, 1H), 7.63 - 7.54 (m, 2H), 7.54 - 7.48 (m, 1H), 7.47 - 7.42 (m, 1H), 7.22 - 7.16 (m, 2H), 7.16 - 7.10 (m, 1H), 7.06 - 6.99 (m, 2H), 6.97 (s, OH), 6.38 (d, J = 7.7 Hz, 1H), 5.66 (d, J = 4.9 Hz, 1H), 5.53 (d, J = 13.8 Hz, 1H), 5.26 (d, J = 13.7 Hz, 1H), 4.76 (t, J = 7.1 Hz, 1H), 4.61 - 4.48 (m, 1H), 4.33 - 4.24 (m, 1H), 4.09 (dt, J = 6.6, 4.7 Hz, 1H), 4.01 (dd, J = 9.1, 6.6 Hz, 1H), 3.84 - 3.77 (m, 1H), 3.57 (ddd, J = 11.9, 7.1, 4.7 Hz, 1H), 3.41 (dt, J = 13.6, 1.0 Hz, 1H), 3.13 (dt, J = 13.7, 1.0 Hz, 1H), 2.47 (s, 3H), 2.41 (s, 3H), 2.17 - 2.08 (m, 1H), 1.26 (d, J = 6.8 Hz, 3H), 0.88 (d, J = 6.6 Hz, 3H), 0.83 (d, J = 6.6 Hz, 3H).

[0358] Example 35: N-(3-(((S)-1-(((S)-1-((1-((2R,4R,5R)-3,3-difluoro-4-hydroxy-5- (hydroxymethyl)tetrahydrofuran-2-yl)-2-oxo-1,2-dihydropyrimidin-4-yl)amino)-1- oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamoyl)phenyl)-2-((3,5- dimethylisoxazol-4-yl)methoxy)-4-nitrobenzamide (Compound 35) Synthesis

[0359] Referring to the synthesis of Example 6, replace intermediate 11a in Example 6 with intermediate 11rv to give a white solid (Compound 35) 1H NMR (600 MHz, DMSO-d6) δ 10.26 (s, 1H), 9.67 (s, 1H), 8.32 (d, J = 9.1 Hz, 1H), 8.22 - 8.14 (m, 2H), 8.10 (t, J = 1.9 Hz, 1H), 7.91 - 7.86 (m, 2H), 7.84 - 7.79 (m, 1H), 7.77 (d, J = 7.6 Hz, 1H), 7.57 (d, J = 9.2 Hz, 1H), 7.46 (t, J = 7.8 Hz, 1H), 6.59 (s, OH), 6.55 (s, OH), 6.52 (s, OH), 6.38 (d, J = 7.7 Hz, 1H), 5.66 (d, J = 4.9 Hz, 1H), 5.46 (d, J = 13.8 Hz, 1H), 5.35 (d, J = 13.8 Hz, 1H), 4.76 (t, J = 7.1 Hz, 1H), 4.61 - 4.48 (m, 1H), 4.34 - 4.26 (m, 1H), 4.16 - 4.09 (m, 2H), 3.79 - 3.71 (m, 1H), 3.64 - 3.56 (m, 1H), 2.37 (s, 3H), 2.31 (s, 3H), 2.28 - 2.19 (m, 1H), 1.30 (d, J = 6.8 Hz, 3H), 0.89 (d, J = 6.7 Hz, 3H), 0.84 (d, J = 6.7 Hz, 3H).

[0360] Example 36: Synthesis of N-(3-(((S)-1-(((S)-1-((5-((2R,3R,4S,5R)-3,4- dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-4-oxo-4,5-dihydro-1,3,5-triazin-2- yl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamoyl)phenyl)-2- ((3,5-dimethylisoxazol-4-yl)methoxy)benzamide (Compound 36)

[0361] (1) Synthesis of tert-butyl (S)-1-(((S)-1-((5-((2R,3R,4S,5R)-3,4-dihydroxy-5- (hydroxymethyl)tetrahydrofuran-2-yl)-4-oxo-4,5-dihydro-1,3,5-triazin-2-yl)amino)-1- oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamate (Intermediate 10rw)

[0362] Referring to the synthetic procedure of Example 1, replace Intermediate Fl in Example 1 with azacitidine to give a white solid (Intermediate 10rw).

[0363] (2) Synthesis of (S)-2-amino-N-((S)-1-((5-((2R,3R,4S,5R)-3,4-dihydroxy-5- (hydroxymethyl)tetrahydrofuran-2-yl)-4-oxo-4,5-dihydro-1,3,5-triazin-2-yl)amino)-1- oxopropan-2-yl)-3-methylbutanamide hydrochloride (Intermediate 11rw)

[0364] Referring to the synthetic procedure of Example 1, tert-butyl ((S)-1-(((S)-1-((5-((2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-4-oxo-4,5- dihydro-1,3,5-triazin-2-yl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2- yl)carbamate was used in place of tert-butyl ((S)-1-(((S)-1-((4-((5-fluoro-2,6-dioxo-3,6- dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxopropan-2-yl)amino)-3- methyl-1-oxobutan-2-yl)carbamate in Example 1 to give a yellow solid (Intermediate 11rw) which was used directly in the next step.

[0365] (3) Synthesis of N-(3-(((S)-1-(((S)-1-((5-((2R,3R,4S,5R)-3,4-dihydroxy-5- (hydroxymethyl)tetrahydrofuran-2-yl)-4-oxo-4,5-dihydro-1,3,5-triazin-2-yl)amino)-1- oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamoyl)phenyl)-2-((3,5- dimethylisoxazol-4-yl)methoxy)benzamide (Compound 36)

[0366] Referring to the synthetic procedure of Example 1, (S)-2-amino-N-((S)-1-((5-((2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-4-oxo-4,5- dihydro-1,3,5-triazin-2-yl)amino)-1-oxopropan-2-yl)-3-methylbutanamide hydrochloride was used in place of (S)-2-amino-N-((S)-1-((4-((5-fluoro-2,6-dioxo-3,6- dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxopropan-2-yl)-3-methylbutanamide hydrochloride in Example 1 to give a white solid (Compound 36) 1H NMR (600 MHz, DMSO-d6) δ 9.64 (s, 1H), 9.40 (s, 1H), 8.32 (d, J = 9.1 Hz, 1H), 7.94 - 7.88 (m, 2H), 7.86 (t, J = 1.9 Hz, 1H), 7.67 - 7.61 (m, 1H), 7.56 (d, J = 9.2 Hz, 1H), 7.53 - 7.48 (m, 1H), 7.45 (t, J = 7.9 Hz, 1H), 7.40 (s, 1H), 7.20 - 7.10 (m, 2H), 5.70 (d, J = 6.1 Hz, 1H), 5.56 (d, J = 13.8 Hz, 1H), 5.36 (d, J = 5.3 Hz, 1H), 5.32 (d, J = 13.7 Hz, 1H), 5.23 (d, J = 5.1 Hz, 1H), 5.05 (t, J = 7.2 Hz, 1H), 4.36 - 4.28 (m, 1H), 4.28 - 4.22 (m, 1H), 3.98 (dt, J = 6.5, 5.4 Hz, 1H), 3.94 - 3.84 (m, 2H), 3.76 - 3.69 (m, 1H), 3.53 - 3.46 (m, 1H), 2.41 (d, J = 19.8 Hz, 6H), 2.14 - 2.02 (m, J = 6.6 Hz, 1H), 1.26 (d, J = 6.8 Hz, 3H), 0.88 (d, J = 6.6 Hz, 3H), 0.83 (d, J = 6.6 Hz, 3H).

[0367] Example 37: Synthesis of N-((S)-l-(((S)-l-((5-((2R,3R,4S,5R)-3,4-dihydroxy-5- (hydroxymethyl)tetrahydrofuran-2-yl)-4-oxo-4,5-dihydro-l,3,5-triazin-2-yl)amino)-l- oxopropan-2-yl)amino)-3-methyl-l-oxobutan-2-yl)-3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamide (Compound 37)

[0368] Referring to the synthesis of Example 13, intermediate 11a in Example 13 was replaced with intermediate 11rw to give a white solid (Compound 37) 1H NMR (600 MHz, DMSO-d6) δ 10.13 (s, 1H), 9.65 (s, 1H), 8.39 (d, J = 9.1 Hz, 1H), 7.90 (dd, J = 7.8, 1.6 Hz, 1H), 7.58 - 7.49 (m, 3H), 7.40 (s, 1H), 7.16 - 7.11 (m, 3H), 7.09 (dd, J = 7.8, 1.3 Hz, 1H), 5.76 (d, J = 5.9 Hz, 1H), 5.44 - 5.34 (m, 2H), 5.28 (d, J = 13.8 Hz, 1H), 5.23 (d, J = 5.1 Hz, 1H), 5.05 (t, J = 7.2 Hz, 1H), 4.37 - 4.28 (m, 1H), 4.02 - 3.93 (m, 2H), 3.91 - 3.84 (m, 1H), 3.81 - 3.69 (m, 5H), 3.53 - 3.46 (m, 1H), 2.39 (d, J = 19.9 Hz, 6H), 2.15 - 2.05 (m, 1H), 1.26 (d, J = 6.8 Hz, 3H), 0.88 (d, J = 6.6 Hz, 3H), 0.83 (d, J = 6.6 Hz, 3H).

[0369] Example 38: 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-(((S)-1-(((S)-1-((5-

[0370] Synthesis of ((2R,4S,5R)-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-4-oxo-4,5- dihydro-1,3,5-triazin-2-yl)amino)-1 -oxopropan-2-yl)aminocarbonyl)phenyl)benzamide (Compound 38)

[0371] (1) Synthesis of tert-butyl ((S)-1-(((S)-1-((5-((2R,4S,5R)-4-hydroxy-5- (hydroxymethyl)tetrahydrofuran-2-yl)-4-oxo-4,5-dihydro-1,3,5-triazin-2-yl)amino)-1 - oxopropan-2-yl)amino)-3-methyl-1 -oxobutan-2-yl)carbamate (Intermediate 10rx)

[0372] Referring to the synthetic procedure of Example 1, Intermediate Fl in Example 1 was replaced with decitabine to give a white solid (Intermediate 10rx).

[0373] (2) Synthesis of (S)-2-amino-N-((S)-1-((5-((2R,4S,5R)-4-hydroxy-5- (hydroxymethyl)tetrahydrofuran-2-yl)-4-oxo-4,5-dihydro-1,3,5-triazin-2-yl)amino)-1- oxopropan-2-yl)-3-methylbutanamide hydrochloride (Intermediate 11rx)

[0374] Referring to the synthetic procedure of Example 1, tert-butyl ((S)-1-(((S)-1-((5- ((2R,4S,5R)-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-4-oxo-4,5-dihydro-1,3,5- triazin-2-yl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamate was used in place of tert-butyl ((S)-1-(((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)- yl)methyl)phenyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamate in Example 1 to give a yellow solid (Intermediate 11rx) which was used directly in the next step.

[0375] (3) 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-(((S)-1-(((S)-1-((5- ((2R,4S,5R)-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-4-oxo-4,5-dihydro-1,3,5- triazin-2-yl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamoyl)phenyl) benzamide (Compound 38)

[0376] ((2R,4S,5R)-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-4-oxo-4,5-dihydro-1,3,5- triazin-2-yl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamoyl)phenyl) benzamide (Compound 38)

[0377] Referring to the synthetic procedure of Example 1, (S)-2-amino-N-((S)-1-((5-((2R,4S,5R)-4- hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-4-oxo-4,5-dihydro-1,3,5-triazin-2-yl)amino)-1- oxopropan-2-yl)-3-methylbutanamide hydrochloride was used in place of (S)-2-amino-N-((S)-1- ((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxopropan-2- yl)-3-methylbutanamide hydrochloride in Example 1 to give a white solid (Compound 38) 1H NMR (600 MHz, DMSO-d6) δ 10.12 (s, 1H), 9.42 (s, 1H), 8.32 (d, J = 9.2 Hz, 1H), 7.94 - 7.88 (m, 2H), 7.74 (t, J = 1.9 Hz, 1H), 7.71 - 7.65 (m, 1H), 7.60 (d, J = 9.2 Hz, 1H), 7.53 - 7.48 (m, 1H), 7.45 (t, J = 7.8 Hz, 1H), 7.40 (s, 1H), 7.20 - 7.14 (m, 1H), 7.12 (dd, J = 7.9, 1.3 Hz, 1H), 6.49 (t, J = 4.4 Hz, 1H), 5.44 (d, J = 13.7 Hz, 1H), 5.29 (d, J = 13.8 Hz, 1H), 5.23 (d, J = 5.4 Hz, 1H), 5.05 (t, J = 7.1 Hz, 1H), 4.29 - 4.20 (m, 2H), 4.15 (dd, J = 9.1, 6.6 Hz, 1H), 3.82 (dt, J = 5.4, 4.6 Hz, 1H), 3.75 - 3.67 (m, 1H), 3.52 - 3.44 (m, 1H), 2.38 (s, 3H), 2.36 - 2.23 (m, 6H), 1.27 (d, J = 6.8 Hz, 3H), 0.88 (d, J = 6.6 Hz, 3H), 0.83 (d, J = 6.6 Hz, 3H).

[0378] Example 39: 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamido)-N-((S)-1-

[0379] Synthesis of (((S)-1-((5-((2R,4S,5R)-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-4-oxo-4,5-dihydro-1,3,5-triazin-2-yl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)-5-methoxybenzamide (Compound 39)

[0380] Reference Example 13 was synthesized by replacing intermediate 11a in Example 13 with intermediate 11rx to give a white solid (Compound 39) 1H NMR (600 MHz, DMSO-d6) δ 10.47 (s, 1H), 9.64 (s, 1H), 8.39 (d, J = 9.1 Hz, 1H), 7.90 (dd, J = 7.8, 1.6 Hz, 1H), 7.56 - 7.46 (m, 2H), 7.40 (s, 1H), 7.36 (t, J = 2.1 Hz, 1H), 7.23 - 7.16 (m, 2H), 7.16 - 7.08 (m, 2H), 6.44 (dd, J = 4.9, 4.0 Hz, 1H), 5.42 (s, 2H), 5.22 (d, J = 5.4 Hz, 1H), 5.05 (t, J = 7.1 Hz, 1H), 4.30 - 4.19 (m, 2H), 4.11 (dd, J = 9.1, 6.6 Hz, 1H), 3.85 - 3.77 (m, 4H), 3.75 - 3.67 (m, 1H), 3.52 - 3.44 (m, 1H), 2.55 (ddd, J = 14.2, 5.6, 4.0 Hz, 1H), 2.42 (s, 3H), 2.31 - 2.21 (m, 1H), 2.04 (s, 3H), 1.86 (dt, J = 14.2, 4.8 Hz, 1H), 1.28 (d, J = 6.8 Hz, 3H), 0.88 (d, J = 6.6 Hz, 3H), 0.83 (d, J = 6.6 Hz, 3H).

[0381] Example 40: Synthesis of 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamide-N-((S)-1-(((S)-1-((4-((5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1- oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)-5-methoxybenzamide (Compound 40)

[0382] (1) Synthesis of tert-butyl ((S)-1-(((S)-1-((4-((5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)- yl)methyl)phenyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamate (Intermediate 10ry)

[0383] Referring to the synthesis method of Example 1, Intermediate F1 in Example 1 was replaced with F2 to obtain a white solid (Intermediate 10ry).

[0384] (2) Synthesis of (S)-2-amino-N-((S)-1-((4-((5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)- yl)methyl)phenyl)amino)-1-oxopropan-2-yl)-3-methylbutanamide hydrochloride (Intermediate 11ry)

[0385] Referring to the synthetic method of Example 1, tert-butyl ((S)-1-(((S)-1-((4-((5-fluoro- 2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxopropan-2-yl)amino)- 3-methyl-1-oxobutan-2-yl)carbamate in Example 1 was replaced with tert-butyl ((S)-1- (((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1- oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamate to give a yellow solid (Intermediate 11ry) which was used directly for the next reaction.

[0386] (3) Synthesis of 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamide-N-((S)-1- (((S)-1-((4-((5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1- oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)-5-methoxybenzamide (Compound 40) Referring to the synthetic method of Example 1, (S)-2-amino-N-((S)-1-((4-((5-fluoro-2,6- dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxopropan-2-yl)-3- methylbutanamide hydrochloride in Example 1 was replaced with (S)-2-amino-N-((S)-1- ((4-((5-fluoro-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1- oxopropan-2-yl)-3-methylbutanamide hydrochloride to give a white solid (Compound 40) 1H NMR (600 MHz, DMSO-d6) δ 11.18 (s, 1H), 10.13 (s, 1H), 9.63 (s, 1H), 8.40 (d, J = 9.1 Hz, 1H), 8.07 (s, OH), 8.04 (s, OH), 7.92 - 7.85 (m, 2H), 7.71 - 7.65 (m, 2H), 7.57 (dt, J = 8.4, 1.0 Hz, 2H), 7.53 - 7.46 (m, 2H), 7.17 - 7.09 (m, 3H), 7.07 (t, J = 2.2 Hz, 1H), 5.40 (d, J = 13.8 Hz, 1H), 5.27 (d, J = 13.8 Hz, 1H), 5.11 (dt, J = 12.2, 1.0 Hz, 1H), 4.91 (dt, J = 12.2, 1.0 Hz, 1H), 4.49 - 4.41 (m, 1H), 4.21 (dd, J = 9.1, 6.6 Hz, 1H), 3.80 (s, 3H), 2.39 (d, J = 15.0 Hz, 5H), 2.10 - 2.00 (m, 1H), 1.25 (d, J = 6.8 Hz, 3H), 0.88 (d, J = 6.6 Hz, 3H), 0.83 (d, J = 6.6 Hz, 3H).

[0387] Example 41: Synthesis of N-(3-(((S)-1-(((S)-1-((9-((2R,3S,4S,5R)-3,4- dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-2-fluoro-9H-purin-6-yl)amino)-1- oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamoyl)phenyl)-2-((3,5- dimethylisoxazol-4-yl)methoxy)benzamide (Compound 41)

[0388] (1) Synthesis of tert-butyl (S)-2-amino-N-((S)-1-((9-((2R,3S,4S,5R)-3,4- dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-2-fluoro-9H-purin-6-yl)amino)-1- oxopropan-2-yl)-3-methylbutanamide (Intermediate 10rz)

[0389] Referring to the synthetic method of Example 1, replace the intermediate F1 in Example 1 with fludarabine to obtain a white solid (Intermediate 10rz).

[0390] (2) Synthesis of (S)-2-amino-N-((S)-1-((9-((2R,3S,4S,5R)-3,4-dihydroxy-5- (hydroxymethyl)tetrahydrofuran-2-yl)-2-fluoro-9H-purin-6-yl)amino)-1- oxopropan-2-yl)-3-methylbutanamide hydrochloride (Intermediate 11rz)

[0391] Referring to the synthetic method of Example 1, tert-butyl ((S)-1-(((S)-1-((9-((2R,3S,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-2-fluoro-9H-purin-6-yl)amino)-1-oxoprop-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamate in Example 1 was replaced with tert-butyl ((S)-1-(((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxoprop-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamate to give a yellow solid (Intermediate 11rz) which was used directly for the next reaction.

[0392] (3) Synthesis of N-(3-(((S)-1-(((S)-1-((9-((2R,3S,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-2-fluoro-9H-purin-6-yl)amino)-1-oxoprop-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamoyl)phenyl)-2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamide (Compound 41) Referring to the synthetic method of Example 1, (S)-2-amino-N-((S)-1-((4-((5-fluoro-2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxoprop-2-yl)-3-methylbutanamide hydrochloride in Example 1 was replaced with (S)-2-amino-N-((S)-1-((9-((2R,3S,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-2-fluoro-9H-purin-6-yl)amino)-1-oxoprop-2-yl)-3-methylbutanamide hydrochloride to give a white solid (Compound 41) 1H NMR (600 MHz, DMSO-d6) δ 9.81 (s, 1H), 9.40 (s, 1H), 8.43 (s, 1H), 8.32 (d, J = 9.1 Hz, 1H), 7.94 - 7.88 (m, 2H), 7.76 - 7.70 (m, 2H), 7.55 - 7.48 (m, 2H), 7.48 - 7.42 (m, 1H), 7.21 - 7.14 (m, 1H), 7.02 (dd, J = 7.9, 1.3 Hz, 1H), 6.10 (d, J = 6.1 Hz, 1H), 5.46 - 5.39 (m, 2H), 5.35 - 5.28 (m, 2H), 5.23 (d, J = 4.4 Hz, 1H), 4.60 - 4.53 (m, 1H), 4.33 - 4.24 (m, 1H), 4.16 (dd, J = 9.1, 6.6 Hz, 1H), 4.12 - 4.07 (m, 1H), 3.98 - 3.92 (m, 1H), 3.81 - 3.74 (m, 1H), 3.60 - 3.52 (m, 1H), 2.40 (d, J = 2.0 Hz, 6H), 2.32 - 2.22 (m, 1H), 1.29 (d, J = 6.8 Hz, 3H), 0.88 (d, J = 6.6 Hz, 3H), 0.83 (d, J = 6.6 Hz, 3H).

[0393] Example 42: N-(3-(((3R,4S,6S,10S,13S,16S)-4-((S)-sec-butyl)-3-(2-((S)-2-

[0394] ((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3- oxopropyl)pyrrolidin-1-yl)-2-oxoethyl)-6,10-diisopropyl-5,11,13,17-tetramethyl-7,9,12,15- tetraoxo-2-oxa-5,8,11,14-tetraazaoctadec-16-yl)carbamoyl)phenyl)-2-((3,5-dimethylisoxazol- 4-yl)methoxy)benzamide (Compound 42)

[0395] (1) ((3R,4S,6S,10S,13S,16S)-4-((S)-sec-butyl)-3-(2-((S)-2-((1R,2R)-3-

[0396] Synthesis of tert-butyl ((3R,4S,6S,10S,13S,16S)-4-((S)-sec-butyl)-3-(2-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1- phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-2-oxoethyl)-6,10-diisopropyl- 5,11,13,17-tetramethyl-7,9,12,15-tetraoxo-2-oxa-5,8,11,14-tetraazaoctadecan-16-yl)carbamate (Intermediate 10st)

[0397] Referring to the synthesis method of Example 1, tert-butyl ((3R,4S,6S,10S,13S,16S)-4-((S)-sec-butyl)-3-(2-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1- phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-2-oxoethyl)-6,10-diisopropyl- 5,11,13,17-tetramethyl-7,9,12,15-tetraoxo-2-oxa-5,8,11,14-tetraazaoctadecan-16-yl)carbamate (Intermediate 10st) was obtained as a white solid.

[0398] (2) Synthesis of (S)-N-(N-L-valyl-L-alanyl-N-methyl-L-valyl)-2-(((3R,4S,5S)-1-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1- phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-3-methoxy-5-methyl-1- oxoheptan-4-yl)(methyl)amino)-3-methylbutanamide hydrochloride (Intermediate 11st)

[0399] Referring to the synthesis method of Example 1, tert-butyl ((3R,4S,6S,10S,13S,16S)-4-((S)-sec-butyl)-3-(2-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1- phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-2-oxoethyl)-6,10-diisopropyl- 5,11,13,17-tetramethyl-7,9,12,15-tetraoxo-2-oxa-5,8,11,14-tetraazaoctadecan-16-yl)carbamate (Intermediate 10st) was obtained as a white solid.

[0400] (3) N-(3-(((3R,4S,6S,10S,13S,16S)-4-((S)-sec-butyl)-3-(2-((S)-2-((1R,2R)-3-(((1S,2R)-1- hydroxy-1-phenylpropan-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-2- oxoehtyl)-6,10-diisopropyl-5,11,13,17-tetramethyl-7,9,12,15-tetraoxo-2-oxa-5,8,11,14- tetraazaoctadecan-16-yl)carbamoyl)phenyl)-2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamide (Compound 42)

[0401] Referring to the synthesis method of Example 1, (S)-2-amino-N-((S)-1-((4-((5-fluoro-2,6- dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxoprop-2-yl)-3- methylbutanamide hydrochloride in Example 1 was replaced with (S)-N-(N-L-valyl-L-alanyl-N- methyl-L-valyl)-2-(((3R,4S,5S)-1-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropan-2- yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidin-1-yl)-3-methoxy-5-methyl-1- oxoheptan-4-yl)(methyl)amino)-3-methylbutanamide hydrochloride to give a white solid (Compound 42) 1H NMR (600 MHz, DMSO-d6) δ 10.02 (s, 1H), 9.40 (s, 1H), 8.32 (d, J = 9.1 Hz, 1H), 8.05 (t, J = 2.2 Hz, 1H), 7.94 - 7.87 (m, 2H), 7.87 - 7.82 (m, 1H), 7.57 - 7.49 (m, 3H), 7.46 (t, J = 7.8 Hz, 1H), 7.34 (d, J = 9.2 Hz, 1H), 7.32 - 7.21 (m, 3H), 7.21 - 7.13 (m, 3H), 5.45 (d, J = 13.8 Hz, 1H), 5.30 (d, J = 13.7 Hz, 1H), 4.77 - 4.70 (m, 1H), 4.59 (d, J = 6.6 Hz, 1H), 4.47 - 4.39 (m, 1H), 4.21 - 4.14 (m, 1H), 4.14 - 4.03 (m, 2H), 3.88 - 3.82 (m, 2H), 3.79 (dd, J = 7.2, 4.3 Hz, 1H), 3.75 - 3.68 (m, 1H), 3.40 - 3.32 (m, 1H), 3.22 (d, J = 19.0 Hz, 6H), 3.13 (d, J = 6.6 Hz, 1H), 2.97 - 2.88 (m, 1H), 2.85 (t, J = 6.2 Hz, 1H), 2.80 - 2.71 (m, 4H), 2.50 (dd, J = 15.8, 6.6 Hz, 1H), 2.45 (s, 3H), 2.37 (s, 3H), 2.28 - 2.13 (m, 5H), 1.99 - 1.86 (m, 2H), 1.86 - 1.71 (m, 3H), 1.63 - 1.54 (m, 1H), 1.46 - 1.36 (m, 1H), 1.36 - 1.26 (m, 4H), 1.23 (d, J = 6.8 Hz, 3H), 1.11 (d, J = 7.1 Hz, 3H), 1.04 - 0.94 (m, 9H), 0.92 - 0.76 (m, 15H).

[0402] Example 43: Synthesis of 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-(((S)-1-(((S)-1-(((2S,3S,4S,6R)-3-hydroxy-2-methyl-6-(((1S,3S)-3,5,12-trihydroxy-3-(2-hydroxyacetyl)-10-methoxy-6,11-dioxo-1,2,3,4,6,11-hexahydrobenzo[e]tetrahydropyran-1-yl)oxy)tetrahydro-2H-pyran-4-yl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamoyl)phenyl)benzamide (Compound 43)

[0403] (1) Synthesis of tert-butyl ((S)-1-(((S)-1-(((2S,3S,4S,6R)-3-hydroxy-2-methyl-6-(((1S,3S)-3,5,12-trihydroxy-3-(2-hydroxyacetyl)-10-methoxy-6,11-dioxo-1,2,3,4,6,11-hexahydro- naphthacene-1-yl)oxy)tetrahydro-2H-pyran-4-yl)amino)-1-oxopropan-2-yl)amino)-3- methyl-1-oxobutan-2-yl)aminocarbamate (Intermediate 10su)

[0404] Referring to the synthesis method of Example 1, Intermediate Fl in Example 1 was replaced with doxorubicin to obtain a white solid (Intermediate 10su).

[0405] (2) Synthesis of (S)-2-amino-N-((S)-1-(((2S,3S,4S,6R)-3-hydroxy-2-methyl-6-(((1S,3S)-3,5,12-trihydroxy-3-(2-hydroxyacetyl)-10-methoxy-6,11-dioxo-1,2,3,4,6,11- hexahydro- naphthacene-1-yl)oxy)tetrahydro-2H-pyran-4-yl)amino)-1-oxopropan-2-yl)-3- methylbutanamide hydrochloride (Intermediate 11su)

[0406] Referring to the synthesis method of Example 1, tert-butyl ((S)-1-(((S)-1-((4-((5-fluoro- 2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxopropan-2-yl)amino)- 3-methyl-1-oxobutan-2-yl)aminocarbamate in Example 1 was replaced with tert-butyl ((S)-1-(((S)-1-(((2S,3S,4S,6R)-3-hydroxy-2-methyl-6-(((1S,3S)-3,5,12-trihydroxy-3-(2- hydroxyacetyl)-10-methoxy-6,11-dioxo-1,2,3,4,6,11-hexahydro- naphthacene-1-yl)oxy) tetrahydro-2H-pyran-4-yl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2- yl)aminocarbamate to obtain a yellow solid (Intermediate 11su), which was used directly in the next reaction.

[0407] (3) 2-((3,5-dimethylisoxazol-4-yl)methoxy)-N-(3-(((S)-1-(((S)-1-(((2S,3S,4S,6R)-3- hydroxy-2-methyl-6-(((1S,3S)-3,5,12-trihydroxy-3-(2-hydroxyacetyl)-10-methoxy-6,11- dioxo-1,2,3,4,6,11-hexahydro- naphthacene-1-yl)oxy)tetrahydro-2H-pyran-4-yl)amino)-1- oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)amino)benzoic acid (Intermediate 12su)

[0408] Synthesis of ((S)-2-amino-N-((S)-1-(((2S,3S,4S,6R)-3-hydroxy-2-methyl-6-(((1S,3S)-3,5,12- trihydroxy-3-(2-hydroxyacetyl)-10-methoxy-6,11-dioxo-1,2,3,4,6,11-hexahydro- naphthacene-1-yl)oxy)tetrahydro-2H-pyran-4-yl)amino)-1-oxopropan-2-yl)-3- methylbutanamide hydrochloride (Compound 43)

[0409] Referring to the synthesis method of Example 1, replacing (S)-2-amino-N-((S)-1-((4-((5-fluoro- 2,6-dioxo-3,6-dihydropyrimidin-1(2H)-yl)methyl)phenyl)amino)-1-oxopropan-2-yl)-3- methylbutanamide hydrochloride in Example 1 with (S)-2-amino-N-((S)-1-(((2S,3S,4S,6R)- 3-hydroxy-2-methyl-6-(((1S,3S)-3,5,12-trihydroxy-3-(2-hydroxyacetyl)-10-methoxy-6,11- dioxo-1,2,3,4,6,11-hexahydro-naphthacene-1-yl)oxy)tetrahydro-2H-pyran-4-yl)amino)-1- oxopropan-2-yl)-3-methylbutanamide hydrochloride, a white solid (Compound 43) was obtained 1H NMR (600 MHz, DMSO-d6) δ 13.34 (s, 1H), 12.38 (s, 1H), 9.40 (s, 1H), 8.32 (d, J = 9.1 Hz, 1H), 7.94 - 7.85 (m, 3H), 7.78 (t, J = 7.8 Hz, 1H), 7.72 - 7.64 (m, 2H), 7.56 - 7.50 (m, 2H), 7.46 (t, J = 7.8 Hz, 1H), 7.23 (d, J = 9.6 Hz, 1H), 7.17 - 7.09 (m, 2H), 7.02 (dd, J = 7.7, 1.1 Hz, 1H), 5.53 (d, J = 13.7 Hz, 1H), 5.40 (dd, J = 6.8, 4.4 Hz, 1H), 5.36 (dd, J = 6.3, 4.0 Hz, 1H), 5.26 (d, J = 13.8 Hz, 1H), 4.73 (s, 1H), 4.59 (t, J = 7.6 Hz, 1H), 4.55 (d, J = 6.9 Hz, 1H), 4.46 (dd, J = 13.7, 7.6 Hz, 1H), 4.23 - 4.08 (m, 3H), 4.03 (dd, J = 9.1, 6.6 Hz, 1H), 3.96 - 3.89 (m, 4H), 3.64 - 3.56 (m, 1H), 3.35 (d, J = 15.6 Hz, 1H), 3.27 (d, J = 15.6 Hz, 1H), 2.50 - 2.43 (m, 4H), 2.33 (dd, J = 13.9, 6.8 Hz, 1H), 2.23 - 2.09 (m, 5H), 2.02 - 1.94 (m, 1H), 1.12 (d, J = 6.9 Hz, 3H), 1.05 (d, J = 6.8 Hz, 3H), 0.88 (d, J = 6.6 Hz, 3H), 0.83 (d, J = 6.6 Hz, 3H).

[0410] Example 44: Synthesis of 3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamide-N-((S)-1-(((S)-1-(((2S,3S,4S,6R)-3-hydroxy-2-methyl-6-(((1S,3S)-3,5,12-trihydroxy-3-(2-hydroxyacetyl)-10-methoxy-6,11-dioxo-1,2,3,4,6,11-hexahydro- tetrazepine-1-yl)oxy)tetrahydro-2H-pyran-4-yl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1- oxobutan-2-yl)-5-methoxybenzamide (Compound 44)

[0411] Referring to the synthesis of Example 13, replace intermediate 11a in Example 13 with intermediate 11su to give a white solid (Compound 44) 1H NMR (600 MHz, DMSO-d6) δ 13.71 (s, 1H), 13.45 (s, 1H), 9.65 (s, 1H), 8.39 (d, J = 9.1 Hz, 1H), 7.90 (dd, J = 8.0, 1.5 Hz, 1H), 7.78 (t, J = 7.8 Hz, 1H), 7.69 (dd, J = 7.8, 1.2 Hz, 1H), 7.64 (t, J = 2.1 Hz, 1H), 7.57 - 7.48 (m, 3H), 7.26 (d, J = 9.6 Hz, 1H), 7.17 - 7.09 (m, 3H), 7.02 (dd, J = 7.7, 1.1 Hz, 1H), 5.48 - 5.39 (m, 2H), 5.37 (dd, J = 6.4, 4.0 Hz, 1H), 5.32 (d, J = 13.7 Hz, 1H), 4.73 (s, 1H), 4.65 (dd, J = 13.5, 7.6 Hz, 1H), 4.59 (t, J = 7.7 Hz, 1H), 4.45 (d, J = 6.9 Hz, 1H), 4.25 (dd, J = 13.5, 7.7 Hz, 1H), 4.23 - 4.11 (m, 3H), 4.02 - 3.95 (m, 1H), 3.92 (s, 3H), 3.78 (s, 3H), 3.73 - 3.64 (m, 1H), 3.39 (d, J = 15.5 Hz, 1H), 3.10 (d, J = 15.6 Hz, 1H), 2.46 - 2.39 (m, 4H), 2.29 - 2.17 (m, 5H), 2.13 - 2.03 (m, 1H), 2.03 - 1.96 (m, 1H), 1.12 (dd, J = 6.8, 3.5 Hz, 6H), 0.88 (d, J = 6.6 Hz, 3H), 0.83 (d, J = 6.6 Hz, 3H).

[0412] Example 45: Synthesis of N-((S)-l-(((S)-l-((9-((2R,3S,4S,5R)-3,4-dihydroxy-5- (hydroxymethyl)tetrahydrofuran-2-yl)-2-fluoro-9H-purin-6-yl)amino)-l-oxopropan-2- yl)amino)-3-methyl-l-oxobutan-2-yl)-3-(2-((3,5-dimethylisoxazol-4-yl)methoxy)benzamide)- 5-methoxybenzamide (Compound 45)

[0413] Referring to the synthesis of Example 13, intermediate 11a in Example 13 was replaced with intermediate 11rz to give a white solid (Compound 45) 1H NMR (600 MHz, DMSO-d6) δ 9.81 (s, 1H), 9.65 (s, 1H), 8.40 (d, J = 9.1 Hz, 2H), 7.90 (dd, J = 7.8, 1.6 Hz, 1H), 7.57 - 7.50 (m, 3H), 7.21 - 7.15 (m, 1H), 7.15 - 7.10 (m, 2H), 7.06 (t, J = 2.2 Hz, 1H), 6.10 (d, J = 6.1 Hz, 1H), 5.42 (d, J = 6.3 Hz, 1H), 5.35 - 5.25 (m, 2H), 5.23 (d, J = 4.4 Hz, 1H), 5.13 (d, J = 13.8 Hz, 1H), 4.62 - 4.55 (m, 1H), 4.14 (dd, J = 9.1, 6.6 Hz, 1H), 4.12 - 4.05 (m, 2H), 3.98 - 3.92 (m, 1H), 3.81 - 3.71 (m, 4H), 3.60 - 3.52 (m, 1H), 2.39 (d, J = 9.6 Hz, 6H), 2.30 - 2.20 (m, 1H), 1.25 (d, J = 6.8 Hz, 3H), 0.89 (d, J = 6.7 Hz, 3H), 0.84 (d, J = 6.7 Hz, 3H).

[0414] Example 46: Evaluation of aldoketoreductase inhibitory activity of compounds of the present application

[0415] 1. Reagents and drugs

[0416] (S)-(+)-1,2,3,4-tetrahydro-1-naphthol (Chiral Technologies Co., Ltd.), oxidized coenzyme II free acid (Shanghai Yuan Ye Biotechnology Co., Ltd.), compounds prepared in Examples 1-30, control compound indomethacin (obtained from Anjieke), AKR1C protein (self-extracted from E. coli BL21 (DE3), plasmid provided by De Tai Biotechnology Co., Ltd.), phosphate buffer (Buffer).

[0417] 2. Instruments

[0418] THERMO Varioskan Flash full-wavelength multifunctional enzyme marker.

[0419] 3. Experimental method

[0420] (1) Preparation of phosphate buffer (Buffer) (pH 7.0): 2.19 g of disodium hydrogen phosphate dodecahydrate and 6.84 g of sodium dihydrogen phosphate dihydrate were dissolved in 470 mL of pure water, and ultrasonic was performed until the solid was completely dissolved. After adjusting the pH to 7.0 with a dilute sodium hydroxide solution, the volume was made up to 500 mL, and it was stored at 4°C.

[0421] (2) Preparation and dilution of 100 mM substrate (S)-(+)-1,2,3,4-tetrahydro-1- naphthalenol: 22.23 mg of substrate was dissolved in 1.5 mL of DMSO to prepare a 100 mM stock solution, vortexed to complete dissolution, and stored in a -20 °C refrigerator for standby. Before each test, the optimal substrate concentration of different subtypes was prepared according to the Km values of AKR1C1-AKR1C4, marked as S1, S2, S3 and S4. Among them, AKR1C1 Km = 107.8 μM, AKR1C2 Km = 184.8 μM, AKR1C3 Km = 106.60 μM, and AKR1C4 Km = 230.45 μM.

[0422] (3) Preparation of 2 mM oxidized coenzyme II free acid NADP+ solution: 12.1 mg of NADP+ was dissolved in 8.138 mL of Buffer. NADP+ solution should be prepared and used immediately, and avoid light.

[0423] (4) Dilution of AKR1C1-AKR1C4: The protein concentration with the maximum response to the substrate was selected for dilution with Buffer, marked as E1, E2, E3 and E4. The extracted AKR1C1 was diluted 80 times, marked as E1; AKR1C2 was diluted 96 times, marked as E2; AKR1C3 was diluted 84 times, marked as E3; and AKR1C4 was diluted 86 times, marked as E4.

[0424] (5) Compound solution gradient dilution: The test compound was prepared and used immediately. The test compound and control compound were prepared into a 100 mM stock solution with DMSO, and Buffer was used for stepwise dilution of the compound: 10 -4 M, 10 -5 M, 10 -6 M, 10 -7 M, 10 -8 M, 10 - 9 M.

[0425] (6) Sample addition: 200 μL of system per well. A total of 8 experimental groups were set up, with 3 replicate wells per group. The compound was set up in 10 -4 M, 10 - 5 M, 10 -6 M, 10 -7 M, 10 -8 M, 10 -9M is 6 test groups, each well is added substrate diluent, NADP+ solution, and different concentrations of compound diluent, Buffer, enzyme diluent. Each experimental group is set a group of negative control group and blank control group, each group of 3 holes. Negative control group is each well is added substrate diluent, NADP+ solution, Buffer, enzyme diluent; blank control group is each well is added substrate diluent, NADP+ solution, Buffer. 37℃ incubation for 10 minutes, after incubation, enzyme labeling instrument in 340nm excitation light and 460nm emission light channel under the determination of each group of fluorescence absorption value. Definition A 测试 is the fluorescence value of test group, A 阴性 is the fluorescence value of negative group, A 空白 is the fluorescence value of blank group, the inhibition rate of each concentration of compound test group is calculated, the formula is as follows: (A 测试 -A 空白 ) / (A 阴性 -A 空白 )×100%. The results obtained by GraphPad PrismTM(GraphPad Software, San Diego, CA, USA) software in non-linear regression analysis model (non-linear regression analysis model) to calculate the corresponding IC 50 value.

[0426] 4、Experimental results

[0427] Table 1. Inhibition activity of target compounds on AKR1C3

[0428]

[0429] * + represents 0-100nM, ++ represents 0.1-1μM, +++ represents 1-10μM, +++++ represents greater than 10μM

[0430] Table 2. Inhibition activity and selectivity of preferred compounds on AKR1C1, AKR1C2, AKR1C3 and AKR1C4

[0431]

[0432] c Selectivity index Selectivity index n (SI n ) = IC 50 AKR1Cn / IC 50 AKR1C3 (n = 1, 2, 4); + represents 0-100nM, ++ represents 0.1-1μM, +++ represents 1-10μM, +++++ represents greater than 10μM

[0433] Results analysis: From the overall results of the activity test, more than half of the 45 compounds had relatively optimal inhibitory activity on AKR1C3. Among them, the IC 50 values of 10 compounds were less than 100 nM, and AKR1C selectivity test was performed, and it was found that these compounds had relatively optimal selectivity on AKR1C3.

[0434] Example 47: Western blotting experiment screened tool cells with high expression of AKR1C3 and cathepsin B (CTSB)

[0435] 1. Experimental materials: A549 human non-small cell lung cancer cells (SCSP-503, China Academy of Sciences Culture Collection Cell Bank), HepG2 human hepatoma cells (SCSP-510, China Academy of Sciences Culture Collection Cell Bank), MDA-MB-436 human breast adenocarcinoma cells (TCHu184, China Academy of Sciences Culture Collection Cell Bank), SH-SY5Y human neuroblastoma cells (SCSP-5014, China Academy of Sciences Culture Collection Cell Bank), U-87MG human brain astrocytoma cells, PC-9 human lung adenocarcinoma cells (provided by Professor Yang Peng's research group), PC-9 EGFR 19DEL / T790M / C797SHuman lung adenocarcinoma cells with EGFR tertiary mutation (provided by Professor Yang Peng's research group), PC-12 rat adrenal pheochromocytoma cells (TCR 8, China Academy of Sciences Typical Culture Collection Cell Library), H460 human large cell lung cancer cells (SCSP-584, China Academy of Sciences Typical Culture Collection Cell Library), MCF-7 / DOX adriamycin-resistant human breast cancer cells (constructed by the research group), PC-3 human prostate cancer cells (SCSP-532, China Academy of Sciences Typical Culture Collection Cell Library), MCF-7 / 1C3 AKR1C3-overexpressing human breast cancer cells (constructed by the research group), DMEM (high sugar) culture medium (containing double antibodies) (KGL1206-500, Kaygen Biotech), special fetal bovine serum (C04001-500, Shanghai Xiaopeng Biological Technology Co., Ltd.), 6-well cell culture plates (703001, NEST), RIPA high-strength lysis solution (Yazhen Biological, PC101), SDS-PAGE protein loading buffer (NCM SDS-PAGE Loading Buffer, 5X) (Xin Saimei, WB2001), 10% color gel rapid kit (P2012, Xin Saimei), trypsin digestion solution (0.25%) (C0201, Biyun Tian), glycine (ST085, Biyun Tian), Tris (ST761, Biyun Tian), sodium dodecyl sulfate (SDS) (ST2681, Biyun Tian), skimmed milk powder (P0216, Biyun Tian), concentrated hydrochloric acid (C0680110228, Nanjing Reagent), BCA quantitative kit (WB6501, Xin Saimei), extremely sensitive ECL chemiluminescence kit (P10060, Xin Saimei), anhydrous ethanol (73537AQ, Adamas), methanol (75851, Adamas), PBS buffer (G4202-500mL, Servicebio), phosphatase inhibitor cocktail (P003, Xin Saimei), HRP-conjugated Goat Anti-Rabbit IgG (H+L) (SA00001-2, Wuhan Sanying), HRP-conjugated Goat Anti-Mouse IgG (H+L) (SA00001-1, Wuhan Sanying), recombinant Anti-AKR1C3 antibody (ab316864, Abeam), PVDF membrane (ISEQ300010, millipore), color pre-stained protein molecular weight marker (P9001, Xin Saimei), Tween 20 (89190EA, Adamas)

[0436] 2. Experimental instruments: Multiskan SkyHigh full-wavelength enzyme label instrument (Thermo Scientific), digital constant-temperature water bath (HH-1, Changzhou Yuxin Instrument Manufacturing Co., Ltd.), MiniProGel TM Protein gel making, electrophoresis and transfer system (E6150, Biyun Tian), PowerPac TM Basic Power Supply (1645050, Bio-Rad), automatic chemical luminescence image analysis system (5200, Tanon), desktop high-speed refrigerated microcentrifuge (D3024R, Dalong), circular shaking table (SLK-O3000-S, SCILOGEX)

[0437] 3. Experimental method:

[0438] a. Cell culture and drug administration: Cells were inoculated in a 6-well plate at a density of 5*103 per well, and incubated at 37°C, 5% CO2, and humidified conditions for 48 h;

[0439] b. Protein extraction: Discard the culture solution, wash 3 times with pre-cooled PBS and discard. Add 60 uL of protein lysis mixture (proteinase inhibitor, phosphatase inhibitor mixture, RIPA lysis buffer = 1:100) to each well, and lyse the cells on ice for 30 min, with slow shaking on a shaking table. Transfer the lysed liquid to a 1.5 mL centrifuge tube and centrifuge for 15 min (4°C, 12000 rpm). Transfer the supernatant to a 1.5 mL centrifuge tube for storage;

[0440] c. Protein concentration determination: Quantify the protein supernatant according to the BCA kit instruction method.

[0441] d. Protein sample denaturation: Add 1 / 4 volume of loading buffer to the protein supernatant, and heat at 95°C for 10 min to fully denature the protein;

[0442] e. Gel preparation: Prepare according to the PAGE gel preparation kit instruction method;

[0443] f. Electrophoresis: After loading the marker and protein, electrophorese in the order of 40V for 15 min, 80V for 60 min, and 120V for 40 min;

[0444] g. Membrane transfer: According to the molecular weight shown by the marker, transfer the gel in the target protein interval to a PVDF membrane using the MiniProGel TM Peel off the gel with a spatula and transfer it to a small transfer sandwich. Carefully attach the PVDF membrane soaked in methanol to the gel. Use the MiniBlot TMBubble roller to remove bubbles. Install the membrane transfer tank sample holder in the following order: black board (the lowest) - sponge pad - filter paper - glue - PVDF membrane - filter paper - sponge pad - white board. Install the electric transfer tank, and place the whole system in ice. Start the membrane transfer at 200 mA for 90 min.

[0445] h. Blocking: Shake the PVDF strips after membrane transfer in 5% skim milk powder at room temperature for 2 h.

[0446] i. Incubation of primary antibody: Prepare the primary antibody solution according to the instructions, and incubate overnight at 4℃.

[0447] j. Incubation of secondary antibody: Take out the strips after overnight incubation of the primary antibody, wash three times with TBST, prepare the secondary antibody solution according to the instructions, and incubate at room temperature for 2 h.

[0448] k. Development: Wash the strips three times with TBST, prepare the ECL luminescent solution according to the instructions, place the strips in the automatic chemiluminescence image analysis system and add the luminescent solution, expose the strips, and analyze the results.

[0449] 4. Experimental results

[0450] From Figure 1 it can be seen that the cells with high expression of AKR1C3 and CTSB are A549, HepG2, PC-9, PC-9EGFR 19DEL / T790M / C797S , H460, MCF-7 / 1C3, which are the positive cell lines screened out.

[0451] Example 48: Evaluation of the effect of the compound on the anti-proliferation of tumor cells

[0452] • 1. Experimental materials: HepG2 human hepatoma cells (SCSP-510, Chinese Academy of Sciences Type Culture Collection Cell Bank), A549 human non-small cell lung cancer cells (SCSP-503, Chinese Academy of Sciences Type Culture Collection Cell Bank), H460 human large cell lung cancer cells (SCSP-584, Chinese Academy of Sciences Type Culture Collection Cell Bank), MCF-7 / 1C3 human breast cancer cells overexpressing AKR1C3 (constructed by the research group), sh-AKR1C3 lentivirus (Jikai Biological, Shanghai, China), cell culture reagents as in Example 47, thiazole blue bromide tetrazolium (JF45956A, Adamas life)

[0453] 2. Experimental method: Cultivate HepG2 cells in a culture medium containing 10% fetal bovine serum, and the culture conditions are 37%, 5% CO2 and saturated humidity.

[0454] (1) Take 5*10 3HepG2(sh-AKRC3) cells were obtained after one week of continuous screening, and Western blotting and qPCR were used for verification;

[0455] (2) HepG2, HepG2(sh-AKRC3), A549, H460, and MCF-7 / 1C3 cells were dispersed in culture medium containing 10% fetal bovine serum and inoculated in 96-well plates at a density of 5*10 3 The 96-well plates seeded with cells were placed in an incubator and incubated at 37°C and 5% CO2 for 18 h until the cells adhered.

[0456] (3) The test compound was gradient-diluted with complete culture medium to the target administration concentration. The original culture medium was discarded, and the cells were treated with blank complete culture medium or 100 μM of the test compound at 37°C and 5% CO2 for 72 h, with 3 replicate wells for each concentration;

[0457] (4) After 72 h, the original culture medium was discarded, and 5 mg / mL thiazolyl blue (prepared in PBS) was added to each well under light shielding conditions, and incubated at 37°C for 4 h. The OD value of each well was detected at 490 nm wavelength by an enzyme-labeled instrument.

[0458] (5) Data processing: cell survival rate = (A 测试 -A 空白 ) / (A 阴性 -A 空白 )*100%. Wherein Ablank is the blank absorption value of the 96-well plate.

[0459] 3. Experimental results

[0460] As shown in Table 3, the classic chemotherapy drug 5-fluorouracil showed a non-selective killing effect on AKR1C3 high / low expression HepG2 cells, while the candidate compounds obtained by screening all showed high anti-proliferation activity on AKR1C3 high expression tumor cells, and showed significantly reduced cytotoxicity on AKR1C3 low expression cells. This result shows that the candidate compounds not only maintain anti-tumor activity, but also show excellent target recognition ability and safety characteristics on tumor cells.

[0461] Table 3. Anti-proliferation activity of 100 μM preferred compounds on tumor cells

[0462]

[0463] * + indicates cell death rate < 20%, ++ indicates cell death rate 20% - 50%, +++ indicates cell death rate 50% - 80%, ++++ indicates cell death rate > 80

Claims

1. A small molecule conjugate, characterized in that, Including compounds of formula (I) or pharmaceutically acceptable salts thereof: in: R1, R2, R3, and R4 are each independently selected from hydrogen, halogen, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 haloalkoxy, C1-C4 alkoxy, nitro, amino, dimethylamino, hydroxy, and cyano. n is selected from 0, 1, and 2; X is selected from L1 is selected from C1-C6 carbon chains and C1-C8 oxygen-containing carbon chains; L2 is selected from any of the following structures: H2N-D is selected from antitumor drugs containing aromatic amine structures and antitumor drugs containing fatty amine structures; -HN-D is formed when the amino group in H2N-D loses a hydrogen atom to become an imino group.

2. The small molecule conjugate according to claim 1, characterized in that, R1, R2, R3, and R4 are each independently selected from hydrogen, fluorine, chlorine, bromine, iodine, methyl, ethyl, propyl, isopropyl, C1-C4 haloalkyl, C1-C4 haloalkoxy, methoxy, ethoxy, nitro, amino, dimethylamino, hydroxy, and cyano. n is selected from 0, 1, and 2; L1 is selected from any of the following structures: -HN-D is selected from any of the following structures:

3. The small molecule conjugate according to claim 1, characterized in that, The compound is selected from any of the following compounds:

4. The small molecule conjugate according to claim 1, characterized in that, The pharmaceutically acceptable salt is a salt formed by the compound with any of the following acids: Hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, carbonic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, citric acid, tartaric acid, malic acid, lactic acid, pyruvic acid, acetic acid, maleic acid, succinic acid, fumaric acid, salicylic acid, phenylacetic acid, mandelic acid, ferulic acid.

5. A method for preparing the small molecule conjugate according to claim 1, characterized in that, Choose from any of the following methods: Method 1: Compounds 1 and 2 undergo a nucleophilic substitution reaction to give intermediate 3, which is then hydrolyzed to give carboxylic acid intermediate 4. 4 undergoes amide condensation with compound 5 to give intermediate 6, which is further hydrolyzed to give intermediate 7. Compounds 8 and 9 undergo amide condensation to give intermediate 10. 10 is deprotected by the Boc protecting group to give intermediate 11. 11 undergoes amide condensation with compound 12 to give intermediate 13. 13 is deprotected by the Boc protecting group to give intermediate 14. Intermediate 7 and intermediate 14 were subjected to amide condensation to obtain the compound of formula I; Method 2: Intermediate 7 and intermediate 11 obtained by the same steps as in Method 1 are subjected to amide condensation to obtain the compound of Formula I; The definitions of R1, R2, R3, R4, L2, and n are as described in claim 1.

6. The method for preparing the small molecule conjugate according to claim 5, characterized in that, Its pharmaceutically acceptable salt is obtained by salting the corresponding acid with compound I.

7. A pharmaceutical composition, characterized in that, It includes the small molecule conjugate drug as described in claim 1 and a pharmaceutically acceptable carrier.

8. The use of a small molecule conjugate drug of claim 1 or a pharmaceutical composition of claim 7 in the preparation of a drug for the prevention or treatment of cancer.

9. The application according to claim 8, characterized in that, The prevention or treatment of cancer includes preventing cancer, treating cancer, or reversing tumor drug resistance.

10. The application according to claim 8, characterized in that, The cancers mentioned include liver cancer, breast cancer, and lung cancer.

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