7-Carbonylstaurosporine Derivatives and Their Preparation and Use

Abstract

The present invention discloses 7-carbonylstaurosporine derivatives and their preparation and use. The present invention further discloses chemical synthesis methods for 18 such representative compounds. The synthetic routes are simple, easy to operate and implement, and the necessary reagents are readily available. The compounds disclosed in the present invention have significant PIM-1 kinase inhibitory activity and excellent kinase selectivity, and can be used in the development of therapeutic agents for diseases or disorders caused by abnormal expression of PIM-1 kinase.

Description

[Technical Field]

[0001] The present invention relates to the pharmaceutical field, specifically to 7-carbonylstaurosporine derivatives, their preparation methods, and their use in the pharmaceutical field. The 7-carbonylstaurosporine derivatives of the present invention can be used as or for the preparation of drugs that selectively inhibit PIM-1 with few side effects, and are particularly suitable for the treatment of diseases or disorders associated with PIM-1 activity. [Background technology]

[0002] PIM kinases belong to the calcium / calmodulin kinase (CAMK) family and have three major subtypes: PIM-1, PIM-2, and PIM-3. Their amino acid sequences are highly homologous: PIM-1 and PIM-2 share 61% similarity, PIM-1 and PIM-3 share 71% similarity, and PIM-2 and PIM-3 share 44% similarity.

[0003] Three subtypes of PIM kinases are expressed in different tissues: PIM-1 is highly expressed in hematopoietic cells, lymphocytes, and prostate cells, PIM-2 is highly expressed in lymphocytes and brain cells, and PIM-3 is highly expressed in breast, kidney, and brain cells.

[0004] Overexpression of PIM kinases not only leads to cancer cell proliferation but also causes cancer cell resistance to conventional treatment strategies, such as chemotherapy, radiotherapy, and the immunosuppressant rapamycin. Interactions between PIM kinases and other oncogenic pathways, such as the PI3K / mTOR / AKT signaling pathway, lead to cancer cell growth, proliferation, and escape from apoptosis. Various cancer-promoting signaling molecules, such as MYC, CDC25, and BAD, are downstream targets of PIM kinases, and their expression is regulated by pathways such as JAK-STAT and NF-κB.

[0005] Due to the relative lack of structural information on PIM-2 and PIM-3, PIM-1 has been the most widely studied member of the PIM kinase family. After more than 20 years of development and research, many PIM-1 kinase inhibitors with various heterocyclic skeletons have been designed, but only a few compounds have entered clinical studies, and to date, no PIM-1 kinase inhibitors have been approved for sale.

[0006] Staurosporine is a broad-spectrum kinase inhibitor and the most representative indolecarbazole compound. Many of its derivatives, such as midostaurin, lestaurtinib, and UCN-01, have already been approved for commercial sale or are in clinical research, demonstrating the great potential of these compounds. However, low kinase selectivity and significant side effects have been major obstacles to their further development.

[0007] The present inventors have investigated the structure-activity relationship of existing indolecarbazole compounds in the inhibitory activity of PIM-1 kinase and found that the carbonyl at C-7 is crucial for PIM-1 kinase inhibition. Therefore, they modified the structure of 7-carbonylstaurosporine to obtain derivatives that exhibit highly selective inhibitory activity against PIM-1 kinase.

[0008] The present invention uses 7-carbonylstaurosporine as a lead compound to obtain a series of derivatives by chemical synthesis. The PIM-1 kinase inhibitory activity of these compounds was measured, and it was found that the 7-carbonylstaurosporine derivatives have excellent inhibitory activity against PIM-1. In particular, compound 17 exhibits a significant inhibitory activity against PIM-1 kinase, and is an IC 50 The IC was 5.9 nM, and the effect was significantly greater than that of the positive control drug TP-3654 (IC 50 Further studies revealed that compound 17 has excellent selective inhibitory activity against PIM-1 kinase and is highly promising for future development. Summary of the Invention [Problem to be solved by the invention]

[0009] The present invention may be useful in developing drugs using such compounds to treat related diseases or disorders caused by the abnormal expression of PIM-1 kinase. [Means for solving the problem]

[0010] The present invention provides a 7-carbonylstaurosporine derivative or a pharmaceutically acceptable salt thereof, wherein the 7-carbonylstaurosporine derivative has a structure represented by the following formula (I): [ka] (I) (In formula (I), L is -(CH2) n -, -(CH2) n NH-, -(CH2) n O-, -(CH2) n C(=O)O-, -(CH2) n C(=O)NH-, -(CH2) n S-, -(CH2) n S(=O)-, -(CH2) n S(=O)2-, -(OCH2CH2O) n -, -(CH2CH2O) n -, -(OCH2CH2OCH2) n -, -(CH2CH2OCH2) n -, -(CH2CH2OCH2CH2) n -, an alkenylene group, an alkynylene group, a cycloalkylene group, a heteroaromatic hydrocarbylene group, or any combination thereof, where n represents a natural number from 1 to 10; R is H, halogen, hydroxyl group, amino group, cyano group, nitro group, C 1~6 Alkyl group, C 2~6 Alkenyl group, C 2~6 Alkynyl group, C 3~10Cyclic hydrocarbon groups, 3- to 10-membered heterocyclic groups, -C(=O)R1, -OC(=O)R1, -C(=O)OR1, -OR1, -SR1, -S(=O)R1, -S(=O)2R1, -S(=O)2NR1R2, -NR1R2, -C(=O)NR1R2, -NR1C(=O)R2, -NR1C(=O)OR2, -NR1S(=O)2R2, -NR1C(=O)NR1R2, -C 1~6 Alkylene-NR1R2, -C 1~6 Alkylene -O(P=O)(OH)2, and -OC 1~6 alkylene-NR1R2; The above alkyl, alkylene, alkenyl, alkynyl, cyclic hydrocarbon, and heterocyclic groups each represent a halogen, a hydroxyl group, an oxo group, an amino group, a cyano group, a nitro group, a C 1~6 Alkyl group, C 2~6 Alkenyl group, C 2~6 Alkynyl group, C 3~6 Cyclic hydrocarbon groups, 3- to 10-membered heterocyclic groups, =N-OR1, -C(=NH)NH2, -C(=O)R1, -OC(=O)R1, -C(=O)OR1, -OR1, -SR1, -S(=O)R1, -S(=O)2R1, -S(=O)2NR1R2, -NR1R2, -C(=O)NR1R2, -NR1C(=O)R2, -NR1C(=O)OR2, -NR1S(=O)2R2, -NR1C(=O)NR1R2, -C 1~6 Alkylene -NR1R2, -OC 1~6 optionally substituted by one or more substituents independently selected from alkylene-NR1R2; R1 and R2 each independently represent H, C, 1~6 Alkyl group, C 3~10 Alternatively, when R1 and R2 are linked to the same nitrogen atom, R1 and R2 together with the atom to which they are linked optionally form a 3- to 12-membered heterocyclic ring.

[0011] Furthermore, the 7-carbonylstaurosporine derivative may be any one selected from the following compounds 1 to 18. [ka]

[0012] The present invention also provides a pharmaceutical composition comprising the 7-carbonylstaurosporine derivative or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable carriers, diluents or excipients.

[0013] The present invention also provides use of the 7-carbonylstaurosporine derivative or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition, in the preparation of a medicament for inhibiting PIM-1 kinase.

[0014] The present invention also provides use of the 7-carbonylstaurosporine derivative or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition, in the preparation of a medicament for treating and / or preventing a disease associated with abnormal activity of PIM-1 kinase.

[0015] The disease may be cancer.

[0016] The cancer is preferably leukemia, lymphoma, multiple myeloma, breast cancer, endometrial cancer, uterine cancer, vaginal cancer, fallopian tube cancer, cervical cancer, kidney cancer, bladder cancer, urothelial cancer, urethral cancer, prostate cancer, testicular cancer, colorectal cancer, sarcoma, bone cancer, myxoma, rhabdomyoma, leiomyoma, fibroma, lipoma, teratoma, pharyngeal cancer, nasopharyngeal cancer, oral cancer, lung cancer, alveolar cancer, mesothelioma, small intestine cancer, gastric cancer, esophageal cancer, pancreatic cancer, liver cancer, bile duct cancer, neurofibroma, glioma, neuroblastoma, melanoma, skin cancer, basal cell carcinoma, squamous cell carcinoma, thyroid cancer, head and neck cancer, salivary gland cancer, or gastrointestinal stromal tumor.

[0017] Preferably, the lymphoma is Hodgkin's disease or non-Hodgkin's lymphoma (e.g., mantle cell lymphoma, diffuse large B-cell lymphoma, follicle center lymphoma, marginal zone B-cell lymphoma, lymphoplasmacytic lymphoma, and peripheral T-cell lymphoma), the lung cancer is non-small cell lung cancer (NSCLC) (including squamous cell carcinoma, adenocarcinoma, and large cell carcinoma, etc.) or small cell carcinoma (SCLC), and the kidney cancer is renal cell carcinoma, clear cell tumor, and renal eosinophilic cell adenoma. oncocytoma), the leukemia is chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia (ALL), T-cell acute lymphoblastic leukemia (T-ALL), chronic myeloid leukemia (CML) or acute myeloid leukemia (AML), the colorectal cancer is colon cancer or rectal cancer, and the sarcoma is chondrosarcoma.

[0018] The present invention further relates to a method for inhibiting PIM-1 kinase, which comprises administering to a patient in need thereof a therapeutically effective amount of the 7-carbonylstaurosporine derivative or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same.

[0019] The present invention further relates to a method for treating and / or preventing a disease associated with abnormal activity of PIM-1, which comprises administering a therapeutically effective amount of the 7-carbonylstaurosporine derivative or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing the same to a patient in need thereof.

[0020] The disease coverage may be the same as above.

[0021] The active compound can be prepared into a form suitable for administration by any suitable route, and the composition of the present invention is prepared by conventional methods using one or more pharmaceutically acceptable carriers.Therefore, the active compound of the present invention can be formulated into various dosage forms for oral administration, injection (for example, intravenous, intramuscular or subcutaneous), inhalation or insufflation administration.The compound of the present invention can also be formulated into dosage forms such as tablets, hard or soft capsules, aqueous or oily suspensions, emulsions, injections, dispersible powders or granules, suppositories, lozenges or syrups.

[0022] As a general guideline, the active compound is preferably in unit dose form or in a form that can be self-administered by the patient in a single dose. Unit doses of the compounds or compositions of the present invention can be provided in the form of tablets, capsules, cachets, bottled liquids, powders, granules, lozenges, suppositories, reconstituted powders, or liquid formulations. Suitable unit doses are 0.1 to 1000 mg.

[0023] The pharmaceutical composition of the present invention may contain one or more auxiliary materials in addition to the active compound. The auxiliary materials may be selected from fillers (diluents), binders, wetting agents, disintegrants, excipients, etc. Depending on the method of administration, the composition may contain 0.1 to 99% by weight of the active compound.

[0024] The tablets contain the active ingredient and non-toxic pharmaceutically acceptable excipients suitable for the preparation of tablets. These excipients may be inert diluents, granulating agents, disintegrating agents, binders, lubricants, etc. The tablets may be uncoated or may be coated by known techniques to mask the taste of the drug or to delay disintegration and absorption in the gastrointestinal tract, thereby providing a sustained-release effect over a long period of time.

[0025] Oral formulations may be provided in soft gelatin capsules in which the active ingredient is mixed with an inert solid diluent, or in which the active ingredient is mixed with a water-soluble carrier or oily medium.

[0026] Aqueous suspensions contain the active substance and mixing excipients suitable for the preparation of an aqueous suspension. Such excipients are suspending agents, dispersing agents, or wetting agents. Aqueous suspensions may also contain one or more preservatives, one or more coloring agents, one or more flavoring agents, and one or more sweetening agents.

[0027] Oily suspensions can be prepared by suspending active ingredients in vegetable oil or mineral oil.Oily suspensions can also contain thickeners.Sweeteners and flavoring agents as mentioned above can be added to provide a smooth and palatable formulation.These compositions can be protected by adding antioxidants.

[0028] The pharmaceutical composition of the present invention may be in the form of an oil-in-water emulsion. The oil phase may be a vegetable oil, a mineral oil, or a mixture thereof. Suitable emulsifiers may be natural phospholipids, and the emulsifier may contain sweeteners, flavoring agents, preservatives, and antioxidants. Such formulations may also contain emollients, preservatives, coloring agents, and antioxidants.

[0029] The pharmaceutical compositions of the present invention may be in the form of a sterile injectable aqueous solution. Acceptable vehicles or solvents that can be used include water, Ringer's solution, and isotonic sodium chloride solution. The sterile injectable preparation may also be a sterile injectable oil-in-water microemulsion in which the active ingredient is dissolved in the oil phase; the injectable solution or microemulsion may be injected into the patient's bloodstream by local bulk injection. Alternatively, solutions and microemulsions are preferably administered to maintain a constant circulating concentration of the compound of the present invention. To maintain such a constant concentration, a continuous intravenous drug delivery device can be used. An example of such a device is the Deltec CADD-PLUS™ 5400 intravenous infusion pump.

[0030] The pharmaceutical compositions disclosed in the present invention may be in the form of sterile injectable aqueous or oily suspensions for intramuscular and subcutaneous administration. Such suspensions can be formulated according to known techniques using the above-mentioned suitable dispersing or wetting agents and suspending agents. Sterile injectable preparations may be sterile injectable solutions or suspensions prepared using non-toxic parenterally acceptable diluents or solvents. In addition, sterile fixed oils can be easily used as solvents or suspending media. For this purpose, any formulated fixed oil can be used. In addition, fatty acids can also be used in the preparation of injectables.

[0031] The compounds of the present invention can be administered in the form of suppositories for rectal administration. These pharmaceutical compositions can be prepared by mixing the drug with a suitable non-irritating excipient that is solid at room temperature but liquid in the rectum and melts in the rectum to release the drug.

[0032] The compounds of the present invention can be administered by adding water to prepare dispersible powders and granules that are suspended in water. These pharmaceutical compositions can be prepared by mixing the active ingredient with a dispersing agent, wetting agent, suspending agent, or one or more preservatives.

[0033] As is well known to those skilled in the art, the dosage of a drug varies depending on various factors, including, but not limited to, the activity of the specific compound used, the severity of the disease, the patient's age, the patient's weight, the patient's health condition, the patient's behavior, the patient's diet, the administration time, the administration method, the excretion rate, the combination of drugs, etc. Furthermore, the optimal treatment regimen, such as the treatment pattern, the daily dosage, and the type of pharmaceutically acceptable salt, can be verified according to conventional treatment regimens.

[0034] The present invention also provides a method for preparing compounds 1 to 18, which uses 7-carbonylstaurosporine as a starting substrate, and the synthesis route is as follows: [ka] Synthesis of Compounds 1-11 by the Acyl Halide Method: The starting substrate and each of the acyl halides having different groups were reacted in a mixture of N,N-diisopropylethylamine (DIPEA) and chloroform, or N,N-diisopropylethylamine and anhydrous N,N-dimethylformamide (DMF), to obtain Compounds 1-11, respectively. Synthesis of compounds 12-15 by the anhydride method: The starting substrate was reacted with each of the different anhydrides in a mixture of 4-dimethylaminopyridine (DMAP) and dimethyl sulfoxide (DMSO) to give compounds 12-15, respectively. Synthesis of compound 16 by onium salt condensation agent method: The starting substrate and suberic acid are reacted in a mixture of N,N-diisopropylethylamine, N,N-dimethylformamide, and O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU) to give compound 16. Synthesis of compound 17 by CDI condensation method: First, compound 12 is reacted with N,N'-carbonyldiimidazole (CDI) and N,N-dimethylformamide in a mixed environment, and then hydroxylamine hydrochloride is added and further reacted to obtain compound 17. Compound 12 is reacted with 3-aminopyrazole in a mixture of N,N-diisopropylethylamine, N,N-dimethylformamide, and O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU) to give compound 18.

[0035] The above preparation method involves a total of one to two steps, making it easy to operate and carry out. [Effects of the Invention]

[0036] Compared with the prior art, the present invention has the following beneficial effects:

[0037] The compounds disclosed in the present invention have significant PIM-1 kinase inhibitory activity and excellent kinase selectivity, and can be used in the development of therapeutic agents for diseases or disorders caused by abnormal expression of PIM-1 kinase.

[0038] The compounds of the present invention have a simple synthetic route, are easy to operate and implement, and the necessary reagents are readily available. Derived compound 17 has a stronger inhibitory effect on PIM-1 kinase than TP-3654, a potent anti-cancer drug currently under clinical development, and has excellent kinase selectivity, making it more advantageous for clinical target-specific therapy. DETAILED DESCRIPTION OF THE INVENTION

[0039] The present invention will be further described below with reference to specific examples. It should be understood that these examples are for the purpose of illustrating the present invention and are not intended to limit the scope of the present invention. In the following examples, the operating methods for which no specific conditions are specified are usually carried out under conventional conditions or conditions recommended by the manufacturer. Example 1. Synthesis of derivatives

[0040] The synthesis steps of compounds 1 to 8 are as follows. 10.0 mg (0.021 mmol) of 7-carbonylstaurosporine was weighed into a 25 mL reaction flask and dissolved in 2 mL of chloroform. Then, 40 μL (0.230 mmol) of DIPEA and 40 μL (0.476 mmol) of propanoyl chloride were added and the mixture was stirred at 30°C for 4 h. The reaction progress was monitored by thin-layer chromatography. After concentration under reduced pressure, the mixture was separated using HPLC. The mobile phase was 85% methanol-water-0.05% TFA, the flow rate was 10 mL / min, and the detection wavelength was 318 nm. Compound 1 (4.0 mg, yield approximately 35.8%, t R =18min). The reactant was replaced with n-butyryl chloride, the mobile phase was changed to 87% methanol-water-0.05% TFA, the flow rate was 10 mL / min, and the detection wavelength was changed to 318 nm. Compound 2 (4.0 mg, yield approximately 34.9%, tR The reactant was replaced with valeryl chloride, the mobile phase was changed to 87% methanol-water-0.05% TFA, the flow rate was changed to 10 mL / min, and the detection wavelength was changed to 318 nm, to obtain Compound 3 (6.1 mg, yield approximately 51.9%, t R The reactant was replaced with capronyl chloride, the mobile phase was changed to 90% methanol-water-0.05% TFA, the flow rate was changed to 10 mL / min, and the detection wavelength was changed to 318 nm, to obtain Compound 4 (8.0 mg, yield: approximately 66.4%, t R The reactant was replaced with heptanoyl chloride, the mobile phase was changed to 92% methanol-water-0.05% TFA, the flow rate was changed to 10 mL / min, and the detection wavelength was changed to 318 nm, to obtain Compound 5 (4.5 mg, yield approximately 36.5%, t R The reactant was replaced with caprylyl chloride, the mobile phase was changed to 93% methanol-water-0.05% TFA, the flow rate was changed to 10 mL / min, and the detection wavelength was changed to 318 nm, to obtain compound 6 (4.9 mg, yield approximately 38.8%, t R The reactant was replaced with cyclopropanecarbonyl chloride, and the mobile phase was changed to 85-100% methanol-water-0.05% TFA. Gradient elution was performed at a flow rate of 10 mL / min and a detection wavelength of 318 nm to obtain compound 7 (4.7 mg, yield approximately 41.2%, t R The reactant was replaced with 2-thiophenecarbonyl chloride, and the mobile phase was changed to 85-100% methanol-water-0.05% TFA. Gradient elution was performed at a flow rate of 10 mL / min and a detection wavelength of 318 nm to obtain compound 8 (5.6 mg, yield approximately 45.6%, t R =19min). The synthesis steps of compounds 9 to 11 are as follows. 10.0 mg (0.021 mmol) of 7-carbonylstaurosporine was weighed into a 25 mL reaction flask and dissolved in 2 mL of anhydrous DMF. 20 μL (0.115 mmol) of DIPEA and 7.8 mg (0.042 mmol) of piperonyl chloride were then added and the mixture was stirred at 30°C for 4 h. The reaction progress was monitored by thin-layer chromatography. After concentration under reduced pressure, the mixture was separated using HPLC with a mobile phase of 68% acetonitrile-water, a flow rate of 10 mL / min, and a detection wavelength of 318 nm. Compound 9 (6.1 mg, yield approximately 46.6%, t R =16min). The reactant was replaced with 3-methylcrotonoyl chloride, the mobile phase was changed to 80% acetonitrile-water-0.05% TFA, the flow rate was 10 mL / min, and the detection wavelength was changed to 318 nm. Compound 10 (6.1 mg, yield approximately 52.1%, t R The reactant was replaced with 3-cinnamoyl chloride, the mobile phase was changed to 80% acetonitrile-water-0.05% TFA, the flow rate was changed to 10 mL / min, and the detection wavelength was changed to 318 nm, to obtain Compound 11 (5.9 mg, yield approximately 46.4%, t R =19min). The synthesis steps of compounds 12 to 15 are as follows. 10.0 mg (0.021 mmol) of 7-carbonylstaurosporine, 3.2 mg (0.032 mmol) of succinic anhydride, and 1.0 mg of DMAP were weighed into a 25 mL reaction flask, dissolved in 3 mL of DMSO, and stirred at 30 °C for 8 h. The reaction progress was monitored by thin-layer chromatography. 50 mL of water was added, and the mixture was extracted three times with an equal volume of ethyl acetate to remove most of the DMSO. The ethyl acetate phase was concentrated under reduced pressure. The mixture was then separated using HPLC with a mobile phase of 52% acetonitrile-water-0.05% TFA, a flow rate of 10 mL / min, and a detection wavelength of 318 nm. Compound 12 (5.5 mg, yield approximately 45.5%, t R =21 min). The reaction mixture was replaced with glutaric anhydride, the mobile phase was changed to 50% acetonitrile-water-0.05% TFA, the flow rate was 10 mL / min, and the detection wavelength was 318 nm. Compound 13 (3.0 mg, yield approximately 24.2%, t R The reactant was replaced with 3-thioglutaric anhydride, the mobile phase was changed to 58% acetonitrile-water-0.05% TFA, the flow rate was changed to 10 mL / min, and the detection wavelength was changed to 318 nm, to obtain Compound 14 (2.8 mg, yield approximately 22.0%, t R The reactant was replaced with adipic anhydride, the mobile phase was changed to 60% acetonitrile-water-0.05% TFA, the flow rate was changed to 10 mL / min, and the detection wavelength was changed to 318 nm to obtain Compound 15 (2.5 mg, yield approximately 19.7%, t R =19min). The synthesis steps of compound 16 are as follows. 7.3 mg (0.042 mmol) of suberic acid and 11.8 mg (0.031 mmol) of HATU were weighed into a 25 mL reaction flask and dissolved in 2 mL of DMF. 20 μL (0.115 mmol) of DIPEA was then added and the mixture was stirred at 30°C for 1 h. Finally, 10.0 mg (0.021 mmol) of 7-carbonylstaurosporine was added and the mixture was stirred at 65°C for 8 h. The progress of the reaction was monitored by thin-layer chromatography. After concentration under reduced pressure, the mixture was separated using HPLC with a mobile phase of 60% acetonitrile-water-0.05% TFA, a flow rate of 10 mL / min, and a detection wavelength of 318 nm. Compound 16 (3.2 mg, yield approximately 24.2%, t R =27 min). The synthesis steps of compound 17 are as follows. 15.0 mg (0.026 mmol) of compound 12 and 8.4 mg (0.052 mmol) of CDI were weighed into a 25 mL reaction flask, dissolved in 2 mL of DMF, and stirred at 30 °C for 1 h. Finally, 7.2 mg (0.104 mmol) of hydroxylamine hydrochloride was added and stirred at 30 °C for 8 h. The reaction progress was monitored by thin-layer chromatography. After concentration under reduced pressure, the mixture was separated using HPLC with a mobile phase of 58% acetonitrile-water-0.05% TFA, a flow rate of 10 mL / min, and a detection wavelength of 318 nm. Compound 17 (2.6 mg, yield approximately 14.0%, t R =11 min). The synthesis steps of compound 18 are as follows. 12.0 mg (0.021 mmol) of compound 12 and 12.2 mg (0.032 mmol) of HATU were weighed into a 25 mL reaction flask and dissolved in 2 mL of DMF. 20 μL (0.115 mmol) of DIPEA was then added and the mixture was stirred at 30°C for 1 h. Finally, 2.0 mg (0.024 mmol) of 3-aminopyrazole was added and the mixture was stirred at 65°C for 8 h. The progress of the reaction was monitored by thin-layer chromatography. After concentration under reduced pressure, the mixture was separated using HPLC with a mobile phase of 65% acetonitrile-water, a flow rate of 10 mL / min, and a detection wavelength of 318 nm. Compound 18 (2.0 mg, yield approximately 12.4%, t R =20 min). Example 2. Identification of Compounds

[0041] Compound 1: Yellow solid. HRESIMS observed m / z 559.1957 [M + Na] + (Calculated value C 31 H 28 N4O5Na, 559.1957). 1H NMR (600 MHz, DMSO-d6): δ 11.14 (s, 1H), 9.26 (d, J = 8.0 Hz, 1H), 9.09 (d, J = 8.0 Hz, 1H), 8.04 (d, J = 8.0 Hz, 1H), 7.73 (d, J = 8.0 Hz, 1H), 7.62 (t, J = 8.0 Hz, 1H), 7.56 (t, J = 8.0 Hz, 1H), 7.43 (t, J = 8.0 Hz, 1H), 7.40 (t, J = 8.0 Hz, 1H), 7.06 (dd, J = 8.8, 6.0 Hz, 1H), 5.07 (ddd, J = 13.1, 4.6, 2.4 Hz, 1H), 4.26 - 4.23 (m, 1H), 2.80 (s, 3H), 2.75 - 2.69 (m, 1H), 2.66 (s, 3H), 2.39 - 2.35 (m, 5H), 2.28 (td, J = 13.1, 6.0 Hz, 1H), 1.05 (t, J = 7.3 Hz, 3H). 13 C NMR (151 MHz, DMSO-d6): δ 173.6, 171.2, 170.9, 139.8, 137.7, 130.6, 128.8, 127.2, 126.9, 125.0, 124.9, 122.8, 121.5, 120.9, 120.7, 120.7, 119.6, 116.1, 115.2, 113.6, 109.9, 95.0, 83.3, 82.5, 60.4, 47.8, 30.7, 29.2, 26.9, 26.4, 9.2. Compound 2: yellow solid. HRESIMS measured value m / z 551.2297 [M + H] + (calculated value C 32 H 31 N4O5, 551.2294). 1H NMR (600 MHz, DMSO-d6): δ 11.13 (s, 1H), 9.26 (d, J = 8.0 Hz, 1H), 9.09 (d, J = 8.0 Hz, 1H), 8.04 (d, J = 8.0 Hz, 1H), 7.73 (d, J = 8.0 Hz, 1H), 7.61 (t, J = 8.0 Hz, 1H), 7.56 (t, J = 8.0 Hz, 1H), 7.45 - 7.38 (m, 2H), 7.05 (dd, J = 8.8, 6.0 Hz, 1H), 5.06 (ddd, J = 13.0, 4.6, 2.4 Hz, 1H), 4.26 - 4.23 (m, 1H), 2.80 (s, 3H), 2.75 - 2.69 (m, 1H), 2.67 (s, 3H), 2.37 (s, 3H), 2.33 (td, J = 7.4, 4.2 Hz, 2H), 2.28 (td, J = 13.0, 6.0 Hz, 1H), 1.58 (h, J = 7.4 Hz, 2H), 0.93 (t, J = 7.4 Hz, 3H). 13 C NMR (151 MHz, DMSO-d6): δ 172.8, 171.2, 170.9, 139.8, 137.7, 130.5, 128.7, 127.2, 126.9, 125.0, 124.9, 122.8, 121.4, 120.9, 120.7, 120.6, 119.6, 116.1, 115.2, 113.6, 109.9, 95.0, 83.3, 82.4, 60.4, 47.8, 35.0, 30.9, 29.2, 26.8, 18.0, 13.9. Compound 3: yellow solid. HRESIMS measured value m / z 565.2447 [M + H] + (calculated value C 33 H 33 N4O5, 565.2451). 1H NMR (600 MHz, DMSO-d6): δ 11.13 (s, 1H), 9.26 (d, J = 8.0 Hz, 1H), 9.08 (d, J = 8.0 Hz, 1H), 8.03 (d, J = 8.0 Hz, 1H), 7.72 (d, J = 8.0 Hz, 1H), 7.61 (t, J = 8.0 Hz, 1H), 7.55 (t, J = 8.0 Hz, 1H), 7.45 - 7.37 (m, 2H), 7.05 (dd, J = 8.8, 6.0 Hz, 1H), 5.05 (ddd, J = 13.1, 4.6, 2.5 Hz, 1H), 4.25 - 4.22 (m, 1H), 2.80 (s, 3H), 2.74 - 2.68 (m, 1H), 2.66 (s, 3H), 2.37 (s, 3H), 2.34 (td, J = 7.4, 2.1 Hz, 2H), 2.27 (td, J = 13.1, 6.0 Hz, 1H), 1.53 (p, J = 7.4 Hz, 2H), 1.37 - 1.29 (m, 2H), 0.89 (t, J = 7.4 Hz, 3H). 13 C NMR (151 MHz, DMSO-d6): δ 173.0, 171.2, 170.9, 139.8, 137.6, 130.5, 128.7, 127.2, 126.9, 125.0, 124.9, 122.8, 121.5, 120.9, 120.7, 120.6, 119.6, 116.1, 115.2, 113.6, 109.8, 95.0, 83.3, 82.4, 60.4, 47.8, 32.8, 30.9, 29.2, 26.8, 26.7, 22.0, 14.0. Compound 4: yellow solid. HRESIMS measured value m / z 579.2612 [M + H] + (calculated value C 34 H 35 N4O5, 579.2607). 1H NMR (600 MHz, DMSO-d6): δ 11.14 (s, 1H), 9.26 (d, J = 8.0 Hz, 1H), 9.08 (d, J = 8.0 Hz, 1H), 8.03 (d, J = 8.0 Hz, 1H), 7.72 (d, J = 8.0 Hz, 1H), 7.61 (t, J = 8.0 Hz, 1H), 7.55 (t, J = 8.0 Hz, 1H), 7.45 - 7.37 (m, 2H), 7.05 (dd, J = 8.8, 6.0 Hz, 1H), 5.05 (ddd, J = 13.1, 4.6, 2.4 Hz, 1H), 4.25 - 4.22 (m, 1H), 2.79 (s, 3H), 2.75 - 2.68 (m, 1H), 2.66 (s, 3H), 2.36 (s, 3H), 2.33 (td, J = 7.2, 1.7 Hz, 2H), 2.27 (td, J = 13.1, 6.0 Hz, 1H), 1.58 - 1.51 (m, 2H), 1.35 - 1.25 (m, 4H), 0.91 - 0.84 (m, 3H). 13 C NMR (151 MHz, DMSO-d6): δ 173.0, 171.2, 170.9, 139.8, 137.6, 130.5, 128.7, 127.2, 126.9, 125.0, 124.9, 122.8, 121.5, 120.9, 120.7, 120.6, 119.6, 116.1, 115.2, 113.6, 109.8, 95.0, 83.3, 82.4, 60.4, 47.8, 33.1, 31.1, 30.9, 29.2, 26.8, 24.3, 22.1, 14.0. Compound 5: yellow solid. HRESIMS measured value m / z 593.2764 [M + H] + (calculated value C 35 H 37 N4O5, 593.2764). 1H NMR (600 MHz, DMSO-d6): δ 11.14 (s, 1H), 9.26 (d, J = 8.0 Hz, 1H), 9.09 (d, J = 8.0 Hz, 1H), 8.04 (d, J = 8.0 Hz, 1H), 7.73 (d, J = 8.0 Hz, 1H), 7.62 (t, J = 8.0 Hz, 1H), 7.56 (t, J = 8.0 Hz, 1H), 7.46 - 7.37 (m, 2H), 7.06 (dd, J = 8.8, 6.1 Hz, 1H), 5.05 (ddd, J = 13.1, 4.6, 2.4 Hz, 1H), 4.26 - 4.22 (m, 1H), 2.80 (s, 3H), 2.75 - 2.69 (m, 1H), 2.68 (s, 3H), 2.37 (s, 3H), 2.34 (td, J = 7.2, 2.0 Hz, 2H), 2.27 (td, J = 13.1, 6.1 Hz, 1H), 1.54 (p, J = 7.2 Hz, 2H), 1.34 - 1.20 (m, 6H), 0.90 - 0.83 (m, 3H). 13 C NMR (151 MHz, DMSO-d6): δ 173.0, 171.2, 170.9, 139.8, 137.7, 130.5, 128.7, 127.2, 126.9, 125.0, 124.9, 122.8, 121.4, 120.9, 120.7, 120.6, 119.6, 116.1, 115.2, 113.6, 109.9, 95.0, 83.3, 82.4, 60.4, 47.8, 33.1, 31.3, 30.9, 29.2, 28.6, 26.8, 24.5, 22.2, 14.1. Compound 6: yellow solid. HRESIMS measured value m / z 607.2926 [M + H] + (calculated value C 36 H 39 N4O5607.2920). 1H NMR (600 MHz, DMSO-d6): δ 11.14 (s, 1H), 9.26 (d, J = 8.0 Hz, 1H), 9.09 (d, J = 8.0 Hz, 1H), 8.03 (d, J = 8.0 Hz, 1H), 7.73 (d, J = 8.0 Hz, 1H), 7.61 (t, J = 8.0 Hz, 1H), 7.56 (t, J = 8.0 Hz, 1H), 7.45 - 7.37 (m, 2H), 7.05 (dd, J = 8.8, 6.1 Hz, 1H), 5.05 (ddd, J = 13.1, 4.6, 2.5 Hz, 1H), 4.26 - 4.21 (m, 1H), 2.80 (s, 3H), 2.74 - 2.68 (m, 1H), 2.67 (s, 3H), 2.37 (s, 3H), 2.35 - 2.31 (m, 2H), 2.27 (td, J = 13.1, 6.1 Hz, 1H), 1.54 (p, J = 7.2 Hz, 2H), 1.31 - 1.22 (m, 8H), 0.88 - 0.82 (m, 3H). 13 C NMR (151 MHz, DMSO-d6): δ 173.0, 171.2, 170.9, 139.8, 137.7, 130.5, 128.7, 127.2, 126.9, 125.0, 124.9, 122.8, 121.4, 120.9, 120.7, 120.6, 119.6, 116.1, 115.2, 113.6, 109.9, 95.0, 83.3, 82.4, 60.4, 47.8, 33.1, 31.3, 30.9, 29.2, 28.9, 28.7, 26.8, 24.6, 22.2, 14.1. Compound 7: yellow solid. HRESIMS measurement value m / z 549.2138 [M + H] + (calculated value C 32 H 29 N4O5549.2138). 1H NMR (600 MHz, DMSO-d6): δ 11.13 (s, 1H), 9.26 (d, J = 8.0 Hz, 1H), 9.09 (d, J = 8.0 Hz, 1H), 8.02 (d, J = 8.0 Hz, 1H), 7.73 (d, J = 8.0 Hz, 1H), 7.62 (t, J = 8.0 Hz, 1H), 7.55 (t, J = 8.0 Hz, 1H), 7.46 - 7.37 (m, 2H), 7.05 (dd, J = 8.8, 6.1 Hz, 1H), 5.02 (ddd, J = 13.0, 4.5, 2.5 Hz, 1H), 4.28 - 4.23 (m, 1H), 2.98 (s, 3H), 2.79 - 2.71 (m, 1H), 2.70 (s, 3H), 2.35 (s, 3H), 2.30 (td, J = 13.0, 6.1 Hz, 1H), 1.99 - 1.91 (m, 1H), 0.93 - 0.73 (m, 4H). 13 C NMR (151 MHz, DMSO-d6): δ 173.3, 171.1, 170.9, 139.9, 137.7, 130.5, 128.7, 127.2, 126.8, 124.9, 124.9, 122.8, 121.4, 120.9, 120.7, 120.6, 119.6, 116.1, 115.2, 113.6, 109.8, 95.0, 83.2, 82.4, 60.4, 48.3, 31.0, 29.2, 26.8, 11.3, 7.5, 7.1. Compound 8: yellow solid. HRESIMS measurement value m / z 591.1694 [M + H] + (calculated value C 33 H 27 N4O5S 591.17O2). 1H NMR (600 MHz, DMSO-d6): δ 11.14 (s, 1H), 9.27 (d, J = 8.0 Hz, 1H), 9.09 (d, J = 8.0 Hz, 1H), 8.06 (d, J = 8.0 Hz, 1H), 7.83 (d, J = 5.0 Hz, 1H), 7.73 (d, J = 8.0 Hz, 1H), 7.65 - 7.54 (m, 3H), 7.46 - 7.38 (m, 2H), 7.18 (s, 1H), 7.13 - 7.03 (m, 1H), 5.01 (s, 1H), 4.48 (s, 1H), 3.06 (s, 3H), 2.96 - 2.87 (m, 1H), 2.68 (s, 3H), 2.46 - 2.27 (m, 4H). 13 C NMR (151 MHz, DMSO-d6): δ 171.1, 170.9, 164.1, 139.8, 137.9, 137.7, 130.6, 130.4, 129.7, 128.8, 127.5, 127.2, 126.9, 125.0, 125.0, 122.8, 121.5, 121.0, 120.7, 120.7, 119.6, 116.1, 115.2, 113.5, 109.8, 95.0, 83.4, 82.4, 60.3, 48.7, 33.6, 29.1, 27.0. Compound 9: orange solid. HRESIMS measured value m / z 629.2032 [M + H] + (calculated value C 36 H 29 N4O7629.2036). 1H NMR (600 MHz, Chloroform-d): δ 9.39 (d, J = 8.0 Hz, 1H), 9.25 (d, J = 8.0 Hz, 1H), 7.77 (d, J = 8.0 Hz, 1H), 7.61 (t, J = 8.0 Hz, 1H), 7.57 - 7.52 (m, 2H), 7.47 (t, J = 8.0 Hz, 1H), 7.44 (t, J = 8.0 Hz, 1H), 7.32 (d, J = 8.0 Hz, 1H), 6.96 (s, 1H), 6.93 (s, 1H), 6.86 (s, 1H), 6.78 (s, 1H), 6.03 (s, 2H), 5.21 (s, 1H), 4.22 (s, 1H), 2.92 (s, 3H), 2.82 (s, 1H), 2.78 - 2.69 (m, 1H), 2.56 (s, 3H), 2.44 (s, 3H). 13 C NMR (151 MHz, Chloroform-d): δ 171.9, 169.9, 169.7, 149.2, 147.8, 139.5, 137.9, 131.6, 130.2, 129.4, 127.2, 127.1, 126.5, 126.3, 123.8, 122.6, 121.9, 121.4, 121.3, 121.0, 119.6, 117.4, 116.7, 116.5, 113.4, 111.7, 108.4, 101.7, 94.9, 84.9, 82.6, 60.5, 49.8, 34.5, 29.0, 28.1. Compound 10: yellow solid. HRESIMS measured value m / z 563.2291 [M + H] + (calculated value C 33 H 31 N4O5563.2294). 1H NMR (600 MHz, DMSO-d6): δ 11.14 (s, 1H), 9.26 (d, J = 8.0 Hz, 1H), 9.09 (d, J = 8.0 Hz, 1H), 8.05 (d, J = 8.0 Hz, 1H), 7.74 (d, J = 8.0 Hz, 1H), 7.62 (t, J = 8.0 Hz, 1H), 7.56 (t, J = 8.0 Hz, 1H), 7.46 - 7.38 (m, 2H), 7.07 (dd, J = 8.7, 6.2 Hz, 1H), 5.97 (s, 1H), 5.09 - 4.98 (m, 1H), 4.31 (s, 1H), 2.82 (s, 3H), 2.79 - 2.72 (m, 1H), 2.70 (s, 3H), 2.39 (s, 3H), 2.29 (td, J = 13.0, 6.2 Hz, 1H), 1.94 (s, 3H), 1.83 (s, 3H). 13 C NMR (151 MHz, DMSO-d6): δ 171.1, 170.9, 167.8, 146.9, 139.9, 137.7, 130.5, 128.7, 127.2, 126.8, 124.9, 124.9, 122.8, 121.4, 120.9, 120.7, 120.7, 119.6, 118.2, 116.1, 115.2, 113.7, 109.9, 95.0, 83.3, 82.4, 60.4, 47.7, 31.6, 29.3, 26.8, 26.1, 20.0. Compound 11: yellow solid. HRESIMS measured value m / z 611.2292 [M + H] + (calculated value C 37 H 31 N4O5611.2294). 1H NMR (600 MHz, DMSO-d6): δ 11.15 (s, 1H), 9.26 (d, J = 8.0 Hz, 1H), 9.10 (d, J = 8.0 Hz, 1H), 8.05 (d, J = 8.0 Hz, 1H), 7.78 - 7.71 (m, 3H), 7.65 - 7.60 (m, 2H), 7.56 (t, J = 8.0 Hz, 1H), 7.47 - 7.37 (m, 5H), 7.25 (d, J = 15.4 Hz, 1H), 7.10 (dd, J = 8.8, 6.1 Hz, 1H), 5.18 (ddd, J = 13.0, 4.7, 2.5 Hz, 1H), 4.36 (s, 1H), 3.03 (s, 3H), 2.83 - 2.78 (m, 1H), 2.70 (s, 3H), 2.41 (s, 3H), 2.37 (td, J = 13.0, 6.1 Hz, 1H). 13 C NMR (151 MHz, DMSO-d6): δ 171.2, 170.9, 166.4, 142.3, 139.8, 137.7, 135.1, 130.6, 129.8, 129.0, 128.9, 128.8, 128.2, 128.2, 127.2, 126.9, 125.0, 124.9, 122.8, 121.5, 120.9, 120.7, 120.7, 119.6, 118.7, 116.1, 115.2, 113.6, 109.9, 95.0, 83.3, 82.4, 60.4, 48.4, 31.2, 29.2, 26.8. Compound 12: yellow solid. HRESIMS measured value m / z 581.2037 [M + H] + (calculated value C 32 H 29 N4O7581.2036). 1H NMR (600 MHz, DMSO-d6): δ 12.09 (s, 1H), 11.14 (s, 1H), 9.26 (d, J = 8.0 Hz, 1H), 9.09 (d, J = 8.0 Hz, 1H), 8.03 (d, J = 8.0 Hz, 1H), 7.75 (d, J = 8.0 Hz, 1H), 7.62 (t, J = 8.0 Hz, 1H), 7.56 (t, J = 8.0 Hz, 1H), 7.45 - 7.38 (m, 2H), 7.09 - 7.04 (m, 1H), 5.01 (ddd, J = 13.1, 4.5, 2.5 Hz, 1H), 4.23 (s, 1H), 2.83 (s, 3H), 2.75 - 2.67 (m, 4H), 2.63 - 2.55 (m, 3H), 2.46 - 2.43 (m, 1H), 2.36 (s, 3H), 2.29 (td, J = 13.1, 6.2 Hz, 1H). 13 C NMR (151 MHz, DMSO-d6): δ 174.1, 172.0, 171.1, 170.9, 139.8, 137.7, 130.5, 128.7, 127.2, 126.9, 125.0, 124.9, 122.8, 121.4, 120.9, 120.7, 120.6, 119.6, 116.1, 115.2, 113.6, 109.8, 95.0, 83.2, 82.4, 60.4, 48.1, 30.8, 29.3, 29.0, 28.4, 26.8. Compound 13: yellow solid. HRESIMS measured value m / z 595.2189 [M + H] + (calculated value C 33 H 31 N4O7595.2193). 1H NMR (600 MHz, DMSO-d6): δ 11.13 (s, 1H), 9.26 (d, J = 8.0 Hz, 1H), 9.09 (d, J = 8.0 Hz, 1H), 8.04 (d, J = 8.0 Hz, 1H), 7.73 (d, J = 8.0 Hz, 1H), 7.62 (t, J = 8.0 Hz, 1H), 7.56 (t, J = 8.0 Hz, 1H), 7.43 (t, J = 8.0 Hz, 1H), 7.40 (t, J = 8.0 Hz, 1H), 7.06 (dd, J = 8.8, 6.1 Hz, 1H), 5.06 (ddd, J = 13.1, 4.5, 2.4 Hz, 1H), 4.26 (s, 1H), 2.80 (s, 3H), 2.76 - 2.70 (m, 1H), 2.68 (s, 3H), 2.42 - 2.38 (m, 2H), 2.38 (s, 3H), 2.31 (t, J = 7.4 Hz, 2H), 2.27 (td, J = 13.1, 6.1 Hz, 1H), 1.78 (p, J = 7.4 Hz, 2H). 13 C NMR (151 MHz, DMSO-d6): δ 174.4, 172.5, 171.2, 170.9, 139.8, 137.6, 130.6, 128.7, 127.2, 126.9, 125.0, 124.9, 122.8, 121.4, 120.9, 120.7, 120.6, 119.6, 116.1, 115.2, 113.6, 109.8, 95.0, 83.3, 82.4, 60.4, 47.8, 33.0, 32.3, 30.8, 29.2, 26.8, 20.1. Compound 14: yellow solid. HRESIMS measured value m / z 613.1761 [M + H] + (calculated value C 32 H 29 N4O7S 613.1757). 1H NMR (600 MHz, DMSO-d6): δ 11.13 (s, 1H), 9.26 (d, J = 8.0 Hz, 1H), 9.09 (d, J = 8.0 Hz, 1H), 8.05 (d, J = 8.0 Hz, 1H), 7.73 (d, J = 8.0 Hz, 1H), 7.62 (t, J = 8.0 Hz, 1H), 7.57 (t, J = 8.0 Hz, 1H), 7.46 - 7.38 (m, 2H), 7.07 (dd, J = 8.8, 6.0 Hz, 1H), 5.00 (ddd, J = 13.0, 4.5, 2.4 Hz, 1H), 4.26 (s, 1H), 3.66 - 3.56 (m, 2H), 3.45 - 3.37 (m, 2H), 2.86 (s, 3H), 2.80 - 2.72 (m, 1H), 2.70 (s, 3H), 2.38 (s, 3H), 2.29 (td, J = 13.0, 6.0 Hz, 1H). 13 C NMR (151 MHz, DMSO-d6): δ 171.2, 171.1, 170.9, 169.1, 139.8, 137.7, 130.5, 128.7, 127.2, 126.9, 125.0, 125.0, 122.7, 121.4, 120.9, 120.7, 120.6, 119.6, 116.1, 115.2, 113.5, 109.9, 95.0, 83.1, 82.4, 60.4, 48.4, 34.2, 33.3, 31.4, 29.2, 26.5. Compound 15: yellow solid. HRESIMS measured value m / z 609.2347 [M + H] + (calculated value C 34 H 33 N4O7609.2349). 1H NMR (600 MHz, Methanol-d4): δ 9.25 (d, J = 8.0 Hz, 1H), 9.04 (d, J = 8.0 Hz, 1H), 7.83 (d, J = 8.0 Hz, 1H), 7.53 - 7.48 (m, 2H), 7.39 (d, J = 8.0 Hz, 1H), 7.34 - 7.29 (m, 2H), 6.76 (dd, J = 9.4, 4.4 Hz, 1H), 5.15 (ddd, J = 13.2, 5.6, 1.8 Hz, 1H), 4.01 (s, 1H), 2.86 (s, 3H), 2.74 - 2.67 (m, 1H), 2.46 - 2.41 (m, 3H), 2.40 (s, 3H), 2.37 (s, 3H), 2.34 (t, J = 6.7 Hz, 2H), 1.71 - 1.63 (m, 4H). 13 C NMR (151 MHz, Methanol-d4): δ 177.2, 175.8, 172.4, 172.2, 140.7, 139.0, 132.4, 130.7, 127.8, 127.7, 126.7, 126.5, 124.6, 123.2, 121.8, 121.6, 121.5, 120.4, 117.9, 116.8, 113.2, 109.7, 96.0, 85.5, 83.8, 60.5, 49.8, 34.5, 34.3, 31.6, 29.1, 28.5, 25.5, 25.4. Compound 16: yellow solid. HRESIMS measured value m / z 637.2671 [M + H] + (calculated value C 36 H 37 N4O7637.2662). 1H NMR (600 MHz, DMSO-d6): δ 11.96 (s, 1H), 11.13 (s, 1H), 9.26 (d, J = 8.0 Hz, 1H), 9.09 (d, J = 8.0 Hz, 1H), 8.04 (d, J = 8.0 Hz, 1H), 7.73 (d, J = 8.0 Hz, 1H), 7.62 (t, J = 8.0 Hz, 1H), 7.56 (t, J = 8.0 Hz, 1H), 7.43 (t, J = 8.0 Hz, 1H), 7.40 (t, J = 8.0 Hz, 1H), 7.06 (dd, J = 8.6, 6.3 Hz, 1H), 5.06 (ddd, J = 13.5, 4.4, 2.4 Hz, 1H), 4.25 (s, 1H), 2.81 (s, 3H), 2.75 - 2.69 (m, 1H), 2.68 (s, 3H), 2.37 (s, 3H), 2.36 - 2.32 (m, 2H), 2.29 - 2.24 (m, 1H), 2.21 (t, J = 7.3 Hz, 2H), 1.59 - 1.53 (m, 2H), 1.53 - 1.47 (m, 2H), 1.35 - 1.25 (m, 4H). 13 C NMR (151 MHz, DMSO-d6): δ 174.6, 172.9, 171.2, 170.9, 139.9, 137.7, 130.5, 128.7, 127.2, 126.9, 125.0, 124.9, 122.8, 121.4, 120.9, 120.7, 120.6, 119.6, 116.1, 115.2, 113.5, 109.8, 95.0, 83.3, 82.4, 60.4, 47.8, 33.7, 33.0, 30.9, 29.2, 28.5, 28.5, 26.8, 24.5, 24.4. Compound 17: yellow solid. HRESIMS measurement value m / z 596.2145 [M + H] + (calculated value C 32 H 30 N5O7596.2145). 1H NMR (600 MHz, DMSO-d6): δ 11.14 (s, 1H), 10.43 (d, J = 1.4 Hz, 1H), 9.26 (d, J = 8.0 Hz, 1H), 9.09 (d, J = 8.0 Hz, 1H), 8.70 (d, J = 1.4 Hz, 1H), 8.03 (d, J = 8.0 Hz, 1H), 7.74 (d, J = 8.0 Hz, 1H), 7.62 (t, J = 8.0 Hz, 1H), 7.56 (t, J = 8.0 Hz, 1H), 7.46 - 7.37 (m, 2H), 7.06 (dd, J = 8.8, 6.0 Hz, 1H), 5.03 (ddd, J = 13.5, 4.6, 2.4 Hz, 1H), 4.24 - 4.18 (m, 1H), 2.82 (s, 3H), 2.76 - 2.66 (m, 4H), 2.64 - 2.55 (m, 3H), 2.36 (s, 3H), 2.32 - 2.25 (m, 2H). 13 C NMR (151 MHz, DMSO-d6): δ 172.0, 171.2, 170.9, 168.7, 139.8, 137.7, 130.6, 128.7, 127.2, 126.9, 125.0, 124.9, 122.8, 121.5, 121.0, 120.7, 120.7, 119.6, 116.1, 115.2, 113.5, 109.9, 95.0, 83.3, 82.4, 60.4, 48.0, 30.8, 29.2, 28.5, 27.4, 26.8. Compound 18: yellow solid. HRESIMS measured value m / z 646.2413 [M + H] + (calculated value C 35 H 32 N7O6646.2414). 1H NMR (600 MHz, Methanol-d4): δ 9.28 (d, J = 8.0 Hz, 1H), 9.08 (d, J = 8.0 Hz, 1H), 7.83 (d, J = 8.0 Hz, 1H), 7.58 - 7.49 (m, 3H), 7.43 (d, J = 8.0 Hz, 1H), 7.34 (t, J = 8.0 Hz, 2H), 6.80 (dd, J = 9.1, 4.3 Hz, 1H), 6.51 (s, 1H), 5.15 (dd, J = 13.4, 5.5 Hz, 1H), 4.07 (s, 1H), 2.93 (s, 3H), 2.81 - 2.70 (m, 5H), 2.50 - 2.45 (m, 1H), 2.45 - 2.43 (m, 3H), 2.40 (s, 3H). 13 C NMR (151 MHz, Methanol-d4): δ 174.8, 172.6, 172.4, 172.3, 154.7, 140.8, 139.1, 132.4, 130.8, 127.9, 127.7, 126.7, 126.5, 124.7, 123.3, 121.9, 121.7, 121.5, 121.1, 120.5, 117.9, 116.9, 113.3, 109.8, 96.1, 92.7, 85.4, 83.9, 60.5, 49.9, 31.7, 31.5, 29.6, 29.2, 28.4. Example 3. Experiments on PIM-1 kinase inhibitory activity and kinase selectivity of compounds

[0042] PIM-1, TRKA, TRKB, FLT3, PKCδ, PKCε, PKCη, PAK4, PLK1, ROCK2, and IKKβ kinase inhibitory activity are all measured using HTRF® KinEASE, a time-resolved fluorescence technology. TM The kinase kit was used for measurement. 7-carbonylstaurosporine (7OSTA) and TP-3654 were used as positive controls, and three wells were set up in parallel. The specific operation steps are as follows: (1) Kinase buffer was prepared and used to prepare test compounds, positive drugs, substrates, kinases, and ATP. (2) To each well of a 384-well microplate, 2 μL of substrate, 2 μL of kinase (no kinase in the negative control group), 4 μL of test compound or positive drug (4 μL of buffer solution in the positive control group, 6 μL of buffer solution in the negative control group), and 2 μL of ATP were added. The microplate was sealed and centrifuged to mix evenly. (3) Depending on the type of kinase, the mixture was incubated in an incubator at 37°C for a certain period of time. (4) A detection solution was prepared using XL-665, antibody, and detection buffer, and 10 μL was added to each well. The microplate was sealed, centrifuged to mix evenly, and then allowed to stand at room temperature for 1 hour. (5) Using a multi-function microplate reader, the fluorescence intensities F1 and F2 were detected at wavelengths of 620 nm and 665 nm, respectively. The signal ratio (F2 / F1 × 10,000) was calculated, and the IC was calculated using Graphpad Prism 8 software. 50 The value was calculated.

[0043] [Table 1]

[0044] [Table 2]

[0045] The results showed that most of the 7-carbonylstaurosporine derivatives synthesized in this invention have excellent inhibitory activity against PIM-1 kinase, and in particular, compound 17 has a significant inhibitory activity against PIM-1 kinase, and the IC 50 The IC was 5.9 nM, and the effect was significantly greater than that of the positive control drug TP-3654 (IC 50The inhibitory activity of compound 17 and 7OSTA against TRKA, TRKB, FLT3, PKCδ, PKCε, PKCη, PAK4, PLK1, ROCK2, and IKKβ kinases was also examined. Compound 17 was found to have excellent selective inhibitory activity against PIM-1 kinase, indicating that this compound can be used for further development of PIM-1 selective inhibitors.

[0046] It should be noted that the terms "comprise," "include," or any variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus comprising a set of elements includes not only those elements but also other elements not expressly listed or inherent to such process, method, article, or apparatus.

[0047] Although preferred embodiments of the present invention have been described, additional changes and modifications can be made to these embodiments by those skilled in the art once they have acquired the basic creative concept. Therefore, it is intended that the appended claims be interpreted to include the preferred embodiments and all changes and modifications that fall within the scope of the present invention.

[0048] It is obvious that those skilled in the art can make various modifications and variations to the present invention without departing from the spirit and scope of the present invention. Therefore, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and its equivalent technologies, the present invention intends to include these modifications and variations.

Claims

1. A 7-carbonylstaurosporine derivative or a pharmaceutically acceptable salt thereof, characterized in that the 7-carbonylstaurosporine derivative is any one selected from the following compounds 4 to 18: 【Chemistry 1】

2. 1. A pharmaceutical composition comprising: A pharmaceutical composition comprising the 7-carbonylstaurosporine derivative or a pharmaceutically acceptable salt thereof according to claim 1, and one or more pharmaceutically acceptable carriers, diluents or excipients.

3. Use of the 7-carbonylstaurosporine derivative or a pharmaceutically acceptable salt thereof according to claim 1, or the pharmaceutical composition according to claim 2, in the preparation of a medicament for inhibiting PIM-1 kinase.

4. Use of the 7-carbonylstaurosporine derivative or a pharmaceutically acceptable salt thereof according to claim 1, or the pharmaceutical composition according to claim 2, in the preparation of a drug for treating and / or preventing a disease associated with abnormal activity of PIM-1 kinase.

5. The use according to claim 4, wherein the disease is cancer.

6. 6. The use of claim 5, wherein the cancer is leukemia, lymphoma, multiple myeloma, breast cancer, endometrial cancer, uterine cancer, vaginal cancer, fallopian tube cancer, cervical cancer, kidney cancer, bladder cancer, urothelial cancer, urethral cancer, prostate cancer, testicular cancer, colorectal cancer, sarcoma, bone cancer, myxoma, rhabdomyoma, leiomyoma, fibroma, lipoma, teratoma, pharyngeal cancer, nasopharyngeal cancer, oral cancer, lung cancer, alveolar carcinoma, mesothelioma, small intestine cancer, gastric cancer, esophageal cancer, pancreatic cancer, liver cancer, bile duct cancer, neurofibroma, glioma, neuroblastoma, melanoma, skin cancer, basal cell carcinoma, squamous cell carcinoma, thyroid cancer, head and neck cancer, salivary gland cancer, or gastrointestinal stromal tumor.

7. 7. The use according to claim 6, wherein the colorectal cancer is colon cancer or rectal cancer.

8. 7. The use according to claim 6, wherein the sarcoma is chondrosarcoma.

9. The method for preparing a 7-carbonylstaurosporine derivative according to claim 1, wherein 7-carbonylstaurosporine is used as a starting substrate, and the synthetic route is as follows: 【Chemistry 2】 Synthesis of Compounds 4 to 11 by the acyl halide method: The starting substrate and each of the acyl halides having different groups were reacted in a mixture of N,N-diisopropylethylamine and chloroform, or N,N-diisopropylethylamine and anhydrous N,N-dimethylformamide to obtain Compounds 4 to 11, respectively. Synthesis of Compounds 12 to 15 by the Anhydride Method: The starting substrate was reacted with each of the different anhydrides in a mixture of 4-dimethylaminopyridine and dimethyl sulfoxide to give Compounds 12 to 15, respectively. Synthesis of Compound 16 by the onium salt condensing agent method: The starting substrate and suberic acid are reacted in a mixed environment of N,N-diisopropylethylamine, N,N-dimethylformamide, and O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate to obtain Compound 16. Synthesis of compound 17 by CDI condensation agent method: First, compound 12 is reacted in a mixed environment of N,N'-carbonyldiimidazole and N,N-dimethylformamide, and then hydroxylamine hydrochloride is added and further reacted to obtain compound 17. A preparation method comprising reacting compound 12 with 3-aminopyrazole in a mixed environment containing N,N-diisopropylethylamine, N,N-dimethylformamide, and O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate to obtain compound 18.

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