Einectin derivative as well as preparation method and application thereof

Abstract

The present invention provides an ecteinin derivative and a preparation method and use thereof, the ecteinin derivative has a structure represented by a general formula (I), and the compound can effectively treat tumor and autoimmune diseases, and can also be applied to antibody-drug conjugates.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology and relates to a sea sucrose derivative, its preparation method, and its uses. Background Technology

[0002] Trabectedin (Et-743) is a structurally complex marine natural product isolated from the marine tunicate *Ecteina scidiaturbinata*, and is the first non-platinum-based novel anti-soft tissue tumor drug. Lubinectedin (PM01183) is a simple synthetic analogue of it. Mechanistic studies have shown that lutentedin covalently binds to guanine residues in the minor groove of DNA to form adducts, leading to double-strand DNA breaks, disrupting DNA-protein interactions and RNA transcription, thereby causing DNA repair impairment, cell cycle disturbances, and cell proliferation arrest or death. Lubinectedin is also an RNA polymerase II inhibitor and can induce the degradation of RNA polymerase II. RNA polymerase II is often overactivated during transcription in tumor cells; lutentedin can cause tumor cells to undergo aberrations and apoptosis during mitosis, ultimately reducing cell proliferation. In addition to its cytotoxic effects, lutentedin can also induce immune cell death and exert synergistic effects with anti-PD-1 or anti-CTLA-4 therapy, targeting CD4 / CD8. + T-cell-dependent methods for treating tumors.

[0003] In June 2020, the U.S. FDA approved rupettedine for patients with small cell lung cancer (SCLC) who had failed first-line platinum-based chemotherapy. In June 2023, the National Medical Products Administration (NMPA) of China accepted the marketing application for rupettedine for the treatment of adult patients with metastatic SCLC whose disease had progressed during or after platinum-based chemotherapy. Previously, this product had been included in the priority review list. In 2023, rupettedine was approved for marketing in Hong Kong and Macau. Furthermore, this product has been introduced as a clinically urgently needed drug to specific medical institutions in the Boao Lecheng International Medical Tourism Pilot Zone in Hainan Province, and is available to Chinese patients through the "Designated Patient Drug Use Program." Using immunotherapy combined with rupettedine after first-line treatment is also a strategy to improve the efficacy of SCLC treatment; currently, several large-scale randomized controlled clinical trials have adopted this treatment modality.

[0004] The FDA approval of rupettedine was based on its Phase II data (2019 ASCO). This study was a single-arm, basket trial of rupettedine as second-line treatment for SCLC, enrolling 105 patients with extensive-stage SCLC who had progressed on prior chemotherapy. All enrolled patients had received platinum-based chemotherapy at 3.2 mg / m². 2Q3W. In terms of efficacy, the ORR was 35.2%, mDOR 5.3 months, mPFS 3.5 months, and mOS 9.3 months. Further subdividing patients with a chemotherapy-free interval (CTFI) ≥90 days (sensitive relapse, n=60) and CTFI <90 days (resistant relapse, n=45), the ORR was 45.0% vs 22.2%, mDOR 6.2 months vs 4.7 months, mPFS 4.6 months vs 2.6 months, and mOS 11.9 months vs 5.0 months, indicating that sensitive patients responded better to rupettedine than resistant patients. Regarding safety, the main grade 3-4 adverse events (whether or not treatment-related) were hematologic toxicity and hepatotoxicity. Grade 3 or higher treatment-related adverse events were mainly febrile neutropenia and fatigue, which were generally manageable.

[0005] On the other hand, cellular experiments have demonstrated that rupettedine, as an RNA polymerase II inhibitor, strongly inhibits cellular RNA synthesis and induces the death of dormant leukemia stem cells. When different concentrations of rupettedine were administered to monocytes in human blood, monocyte apoptosis was closely correlated with drug concentration. The experimental data presented in this study show that rupettedine can effectively kill B lymphocytes and T lymphocytes in human peripheral blood. Therefore, rupettedine and its derivatives have great potential in the treatment of autoimmune diseases.

[0006] As a jugating compound, rubitidin has achieved positive results in clinical chemotherapy applications. However, it remains important to find novel jugating compounds to enhance tumor-killing effects, reduce systemic toxicity, explore new indications, and create new linking sites in antibody-drug conjugates. Summary of the Invention

[0007] To address the shortcomings of existing technologies, the present invention aims to provide a sucrose derivative, its preparation method, and its uses.

[0008] To achieve this objective, the present invention adopts the following technical solution:

[0009] In a first aspect, the present invention provides a sucrose derivative or its tautomer, stereoisomer, or pharmaceutically usable salt thereof, said sucrose derivative having a structure as shown in general formula (I):

[0010]

[0011] Wherein: R1 is selected from cyano, hydroxyl, halogen, carboxyl, R2 is selected from the following groups:

[0012]

[0013] R3 is selected from hydroxyl, carboxyl, The mercapto or 2-mercaptoacetamido group, R4, R5, R6, R7 and R8 are independently selected from hydrogen, C1-C5 alkyl or hydroxyl-substituted C1-C5 alkyl groups; the wavy line represents the linking site of the group;

[0014] m and n are each independently selected from 1, 2, 3 or 4.

[0015] Preferably, R2 is selected from the following groups:

[0016]

[0017] R3 is selected from hydroxyl, carboxyl, and... The mercapto or 2-mercaptoacetamido group, R4, R7 and R8 are independently selected from hydrogen, C1-C5 alkyl or hydroxy-substituted C1-C5 alkyl groups, and the wavy line represents the linking site of the group.

[0018] Preferably, R3 is selected from hydroxyl, amino, carboxyl, methylamino, ethylamino, hydroxyethylamino, mercapto, or 2-mercaptoacetamino.

[0019] Preferably, R4 is selected from methyl, ethyl, isopropyl, or hydroxyethyl.

[0020] More preferably, R2 is selected from the following groups:

[0021]

[0022] The wavy line represents the connection site of the functional group.

[0023] Preferably, the succarpine derivative is selected from any one of the following compounds:

[0024]

[0025]

[0026] In a second aspect, the present invention provides a pharmaceutical composition comprising at least one of the sucrose derivatives or their tautomers, stereoisomers or pharmaceutically acceptable salts as described above, and a pharmaceutically acceptable carrier.

[0027] Thirdly, the present invention also provides an antibody-drug conjugate, wherein the small molecule drug portion of the antibody-drug conjugate includes the sucrose derivatives or their tautomers or stereoisomers as described above; the sucrose derivatives are coupled to an antibody or an antigen-binding fragment of the antibody using a linker to form the antibody-drug conjugate.

[0028] Fourthly, the present invention provides the use of the as-described seasqualin derivatives or their tautomers, stereoisomers or pharmaceutically usable salts in the preparation of medicaments for the treatment and / or prevention of tumors and / or autoimmune diseases.

[0029] Preferably, the diseases include non-small cell lung cancer, small cell lung cancer, colorectal cancer, breast cancer, pancreatic cancer, sarcoma, ovarian cancer, prostate cancer, gastric cancer, liver cancer, kidney cancer, and hematologic malignancies, as well as psoriasis, psoriatic arthritis, systemic lupus erythematosus, lupus nephritis, rheumatoid arthritis, multiple sclerosis, type I diabetes, anti-glomerular basement membrane antibody nephritis / Crohn's disease, ulcerative colitis, myasthenia gravis, vitiligo, etc.

[0030] Compared with the prior art, the present invention has the following beneficial effects:

[0031] This invention provides a seasqualin compound with a specific structure that exhibits better inhibitory activity against a variety of tumor cells and monocytes (lymphocytes), enabling effective treatment of tumors and / or autoimmune diseases, and can also be used in antibody-drug conjugates. Detailed Implementation

[0032] The technical solution of the present invention will be further illustrated below through specific embodiments. Those skilled in the art should understand that the embodiments described are merely illustrative of the present invention and should not be construed as limiting the invention in any way.

[0033] As used herein, the terms "pharmaceutically acceptable salt" or "medicinal salt" refer to salts that enable the compounds of the present invention to retain their original biological activity and are suitable for pharmaceutical use. The pharmaceutically acceptable salt of the compounds represented by formula (I) can be a salt formed by a carboxyl or amino group (primarily an amino group) with a suitable base or acid, such as a salt formed with a suitable base (including metal salts and ammonium salts), or a salt formed with a suitable acid. The compounds of the present invention preferably form salts with a suitable acid, which may be selected from: hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, phosphoric acid, nitric acid, formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, methanesulfonic acid, ethanesulfonic acid, toluenesulfonic acid, benzenesulfonic acid, naphthalenesulfonic acid, trifluoroacetic acid, and aspartic acid, preferably methanesulfonic acid or hydrochloric acid.

[0034] The term “treatment” as used in this article includes any effect that results in improvement of a condition, disease, disorder, etc., such as reducing, decreasing, regulating, improving or eliminating, or improving its symptoms.

[0035] As used herein, the phrase "medicinal carrier" refers to a pharmaceutical substance, composition, or medium, such as a liquid or solid filler, diluent, excipient, manufacturing aid (e.g., lubricant, talc, magnesium stearate, calcium stearate, zinc stearate, or stearic acid), or solvent encapsulation substance, which involves carrying or delivering a main compound from one organ or part of the body to another. Each carrier must be "acceptable" in the sense of compatibility with other components of the formulation and harmlessness to the patient.

[0036] The term "pharmaceutical composition" means a composition comprising the compounds of the present invention and at least one other pharmaceutically acceptable carrier.

[0037] The term "TLC" refers to thin-layer chromatography; the term "NPC" refers to bis(4-nitrophenyl) carbonate; "DCE" is 1,2-dichloroethane; "DMP" is Dess-Martin oxidant; "Ru-Phos-Pd-G3" is (2-dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl)methanesulfonic acid (2-amino-1,1'-biphenyl-2-yl)palladium(II);

[0038] The term "halogen" includes fluorine, chlorine, bromine, and iodine.

[0039] To better illustrate the technical means and effects of the present invention, the present invention will be further described below in conjunction with non-limiting embodiments. The embodiments of the present invention, including the descriptions provided in the embodiments, are intended to illustrate the implementation of the present invention and are not intended to limit the scope of protection of the present invention in any way. According to the present invention, those skilled in the art will understand that many changes can be made to the specific embodiments disclosed without departing from the spirit and scope of the present invention and still obtain the same or similar results.

[0040] Unless otherwise stated, all materials / reagents were obtained from commercial suppliers and were ready for use without further purification. The structures of the compounds in the following examples were characterized and determined by nuclear magnetic resonance (NMR) and / or liquid chromatography-mass spectrometry (LC-MS).

[0041] 1 ¹H NMR spectra were recorded at room temperature using a Bruker Avance 400 MHz spectrometer. The solvents used for analysis were deuterated dimethyl sulfoxide (d-DMSO), deuterated chloroform (CDCl₃), deuterated methanol (CD₃OD), or deuterated water (D₂O). Chemical shift values ​​(δ) are expressed in ppm. Tetramethylsilane (TMS) or the residual solvent peak was used as an internal standard. Coupling constants (J) are expressed in Hertz (Hz). 1The abbreviations for the multiplicity of peaks in HNMR spectra are as follows: s (single peak), d (doublet), t (triplet), q (quartet), qn (quintet), m (multiplet), br (broad peak).

[0042] The instrument used for liquid chromatography-mass spectrometry (LC-MS) was a Shimadzu LCMS-2020, and the instrument used for preparative high-performance liquid chromatography (Prep-HPLC) was a Bonna-Agela FLEXA FL-H100G. The silica gel plates used for thin-layer chromatography (TLC) were Yantai Huanghai HSGF254 TLC plates, with a 2.5×8cm size for reaction monitoring and a coating thickness of 0.2±0.03mm. The plates used for separation and purification were 20×20cm with a coating thickness of 0.4-0.5mm. For silica gel column chromatography, Qingdao Haiyang silica gel (100-200 mesh or 200-300 mesh) was used as the carrier.

[0043] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as are familiar to those skilled in the art. Furthermore, any methods and materials similar to or equivalent to those described herein may be used in the methods of this invention.

[0044] Example 1.1:

[0045] (1'R,6R,6aR,7R,13S,14R,16R)-14-cyano-6'-(3-hydroxyazacyclobutane-1-yl)-8-hydroxy-9-methoxy-4,10,23-trimethyl-19-oxo-2',3',4',6,7,9',12,13,14,16-decahydro-6aH-spiro[7,13-imino-6,16-(cyclothiopropoxymethylene)[1,3]dioxapentano[7,8]isoquinolino[3,2-b][3]benzozocin-20,1'-pyridino[3,4-b]indole]-5-ylacetate

[0046]

[0047] Implementation method:

[0048]

[0049] Step 1: Synthesis of compound 2-(5-chloro-1H-indol-3-yl)ethyl)tert-butyl carbamate

[0050]

[0051] Di-tert-butyl dicarbonate (567 mg, 2.60 mmol) was added dropwise to a methanol (10 mL) solution of 2-(5-chloro-1H-indol-3-yl)ethane-1-amine hydrochloride (1.1-a, 600 mg, 2.60 mmol) and triethylamine (525 mg, 5.20 mmol) at room temperature. The mixture was stirred at room temperature for 0.5 h. Stirring was stopped after complete conversion was monitored by TLC. The reaction solution was concentrated under reduced pressure and diluted with ethyl acetate (10 mL) and water (10 mL). The organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give crude tert-butyl 2-(5-chloro-1H-indol-3-yl)ethyl)carbamate as a pale yellow solid (1.1-b, 730 mg, 95%).

[0052] LC-MS:(ES + ):m / z 239.1[M-55] + .

[0053] Step 2: Synthesis of tert-butyl carbamate (2-(5-(3-hydroxyazacyclobutane-1-yl)-1H-indol-3-yl)ethyl)carbamate

[0054]

[0055] To a suspension of intermediate 2-(5-chloro-1H-indol-3-yl)ethyl)carbamate tert-butyl ester (1,1-b, 500 mg, 1.70 mmol), azacyclobutane-3-ol hydrochloride (929 mg, 8.48 mmol), and cesium carbonate (2.76 g, 8.48 mmol) in 1,4-dioxane (1 mL), Ru-Phos-Pd-G3 (71 mg, 0.085 mmol) was added. The reaction mixture was purged with nitrogen three times and heated to 100 °C with stirring for 14 hours. The reaction mixture was filtered and concentrated under reduced pressure. The residue was diluted with ethyl acetate (10 mL) and water (10 mL), and the organic phase was collected, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (using petroleum ether elution solution containing 75% ethyl acetate) to give tert-butyl 2-(5-(3-hydroxyazacyclobutane-1-yl)-1H-indol-3-yl)ethyl)carbamate as a white solid (1.1-c, 66 mg, 12%).

[0056] LC-MS:(ES + ):m / z 276.2[M-55] + .

[0057] Step 3: Synthesis of compound 1-(3-(2-aminoethyl)-1H-indol-5-yl)azacyclobutane-3-ol trifluoroacetate

[0058]

[0059] Trifluoroacetic acid (1 mL) was added to dichloromethane (2 mL) containing the intermediate (2-(5-(3-hydroxyazacyclobutan-1-yl)-1H-indol-3-yl)ethyl)carbamate (1.1-d, 66 mg, 0.20 mmol). The mixture was reacted at room temperature for 0.5 h. After the reaction was completed by TLC monitoring, stirring was stopped. Water (5 mL) was added to the reaction solution, and the mixture was washed with dichloromethane (5 mL × 2). The aqueous phase was purified by preparative HPLC (mobile phase was 5%–90% aqueous acetonitrile solution, aqueous phase containing 0.1% (v / v) formic acid) to give 1-(3-(2-aminoethyl)-1H-indol-5-yl)azacyclobutan-3-ol trifluoroacetate as a white solid (1.1-d, 45 mg, 69%).

[0060] LC-MS:(ES + ):m / z 232.1[M+H] + .

[0061] Step 4: Synthesis of Compound 1.1: (1'R,6R,6aR,7R,13S,14R,16R)-14-cyano-6'-(3-hydroxyazacyclobutane-1-yl)-8-hydroxy-9-methoxy-4,10,23-trimethyl-19-oxo-2',3',4',6,7,9',12,13,14,16-decahydro-6aH-spiro[7,13-imino-6,16-(cyclothiopropoxymethylene)[1,3]dioxapentano[7,8]isoquinolino[3,2-b][3]benzozocin-20,1'-pyridino[3,4-b]indole]-5-yl acetate

[0062]

[0063] Intermediate Ia (synthetic method according to US5721362) (20 mg, 0.032 mmol) was added to a 2 mL acetic acid solution of intermediate 1-(3-(2-aminoethyl)-1H-indol-5-yl)azacyclobutane-3-ol trifluoroacetate (1.1-d, 32 mg, 0.097 mmol). The mixture was reacted at room temperature for 24 hours. After the reaction was completed by TLC monitoring, stirring was stopped. The reaction solution was concentrated under reduced pressure, diluted with acetonitrile (1 mL) and water (2 mL), and purified by preparative HPLC (mobile phase: 10%–95% aqueous acetonitrile solution, aqueous phase containing 0.1% (v / v) formic acid) to give compound 1.1 (21 mg, 78%).

[0064] LC-MS:(ES + ):m / z 835.1[M+H] + .

[0065] 1 H NMR (400MHz, CDCl3) δ7.58(s,1H),7.11(d,J=8.5Hz,1H),6.65(s,1H),6.42(d,J =2.1Hz,1H),6.38(dd,J=8.6,2.3Hz,1H),6.21(d,J=1.4Hz,1H),6.02(d,J=1.4Hz ,1H),5.84(s,1H),5.13–5.02(m,1H),4.71–4.63(m,1H),4.54(s,1H),4.31(s,1 H),4.27(dd,J=4.6,1.5Hz,1H),4.21(d,J=2.6Hz,1H),4.17(dd,J=11.7,1.9Hz,1 H),4.12(dd,J=7.9,6.1Hz,2H),3.81(s,3H),3.57(dd,J=7.8,4.8Hz,2H),3.42( t,J=7.3Hz,2H),3.15(ddd,J=12.1,7.7,4.8Hz,1H),3.05(d,J=18.1Hz,1H),2.92 (dd,J=18.1,9.0Hz,1H),2.81(dt,J=10.9,5.1Hz,1H),2.59(dt,J=15.1,4.8Hz,1 H),2.55–2.44(m,2H),2.37(s,3H),2.29-2.22(m,4H),2.21(s,3H),2.06(s,3H).

[0066] Example 1.2:

[0067] (1'R,6R,6aR,7R,13S,14R,16R)-14-cyano-8-hydroxy-9-methoxy-6'-(2-hydroxyethoxy)-4,10,23-trimethyl-19-oxo-2',3',4',6,7,9',12,13,14,16-decahydro-6aH-spiro[7,13-imino-6,16-(cyclothiopropoxymethylene)[1,3]dioxapentanoc[7,8]isoquinolino[3,2-b][3]benzozocin-20,1'-pyridino[3,4-b]indole]-5-ylacetate

[0068]

[0069] Implementation method:

[0070]

[0071] Step 1: Synthesis of compound 2-(5-hydroxy-1H-indol-3-yl)ethyl)tert-butyl carbamate

[0072]

[0073] Di-tert-butyl dicarbonate (1.54 g, 7.05 mmol) was added dropwise to a methanol (15 mL) solution of 2-(5-hydroxy-1H-indol-3-yl)ethane-1-amine hydrochloride (1.2-a, 1.50 g, 7.05 mmol) and triethylamine (1.43 g, 14.10 mmol) at room temperature. The mixture was stirred for 0.5 h at room temperature. Stirring was stopped after complete conversion was monitored by TLC. The reaction solution was concentrated under reduced pressure and diluted with ethyl acetate (15 mL) and water (15 mL). The organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give crude tert-butyl 2-(5-hydroxy-1H-indol-3-yl)ethyl)carbamate as a gray solid (1.2-b, 1.73 g, 89%).

[0074] LC-MS:(ES + ):m / z 221.1[M-55] + .

[0075] Step 2: Synthesis of compound ((2-(5-(2-hydroxyethoxy)-1H-indol-3-yl)ethyl)carbamate tert-butyl ester)

[0076]

[0077] Potassium carbonate (1.18 g, 3.62 mmol) was added to a solution of intermediate 2-(5-hydroxy-1H-indol-3-yl)ethyl)carbamate tert-butyl ester (1.2-b, 750 mg, 2.72 mmol) and 2-bromoethanol (1.70 g, 13.58 mmol) in dichloromethane (20 mL) at room temperature. The mixture was stirred under reflux overnight. Stirring was stopped after complete conversion was monitored by TLC. After the mixture was brought back to room temperature, it was diluted with ethyl acetate (15 mL) and water (15 mL). The organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (using dichloromethane elution solution containing 6% methanol) to give the product ((2-(5-(2-hydroxyethoxy)-1H-indol-3-yl)ethyl)carbamate tert-butyl ester as a pale purple oil (1.2-c, 580 mg, 67%).

[0078] LC-MS:(ES + ):m / z 265.0[M-55] + .

[0079] Step 3: Synthesis of compound 2-((3-(2-aminoethyl)-1H-indol-5-yl)oxy)ethane-1-ol trifluoroacetate

[0080]

[0081] Trifluoroacetic acid (1 mL) was added to dichloromethane (2 mL) containing the intermediate ((2-(5-(2-hydroxyethoxy)-1H-indol-3-yl)ethyl)carbamate (1.2-d, 190 mg, 0.60 mmol), and the mixture was reacted at room temperature for 0.5 h. After the reaction was completed by TLC monitoring, stirring was stopped. Water (5 mL) was added to the reaction solution, and the mixture was washed with dichloromethane (5 mL × 2). The aqueous phase was purified by preparative HPLC (mobile phase was 5%–90% acetonitrile aqueous solution, aqueous phase containing 0.5% (v / v) trifluoroacetic acid) to give the product 2-((3-(2-aminoethyl)-1H-indol-5-yl)oxy)ethane-1-ol trifluoroacetate as a white solid (1.2-d, 165 mg, 75%).

[0082] LC-MS:(ES + ):m / z 221.0[M+H] + .

[0083] Step 4: Synthesis of Compound 1.2: (1'R,6R,6aR,7R,13S,14R,16R)-14-cyano-8-hydroxy-9-methoxy-6'-(2-hydroxyethoxy)-4,10,23-trimethyl-19-oxo-2',3',4',6,7,9',12,13,14,16-decahydro-6aH-spiro[7,13-imino-6,16-(cyclothiopropoxymethylene)[1,3]dioxapentanocene[7,8]isoquinolino[3,2-b][3]benzozocin-20,1'-pyridino[3,4-b]indole]-5-yl acetate

[0084]

[0085] Intermediate Ia (20 mg, 0.032 mmol) was added to a 2 mL acetic acid solution of intermediate 2-((3-(2-aminoethyl)-1H-indol-5-yl)oxy)ethane-1-ol trifluoroacetate (1.2-d, 32 mg, 0.097 mmol). The mixture was reacted at room temperature for 24 hours. Stirring was stopped after the reaction was completed by TLC monitoring. The reaction solution was concentrated under reduced pressure, diluted with acetonitrile (1 mL) and water (2 mL), and purified by preparative HPLC (mobile phase: 10%–95% aqueous acetonitrile solution, aqueous phase containing 0.1% (v / v) formic acid) to give compound 1.2 as a white solid (16 mg, 62%).

[0086] LC-MS:(ES + ):m / z 824.3[M+H] + .

[0087] 1 H NMR (400MHz, CDCl3) δ7.64(s,1H),7.15(d,J=8.8Hz,1H),6.85(d,J=2.3Hz,1H),6.78(dd,J=8.8,2.4Hz,1H),6.66(s,1H),6.22(d,J=1.0Hz,1H),6.03( s,1H),5.79(s,1H),5.09(d,J=11.7Hz,1H),4.56(s,1H),4.33(s,1H),4.28 (d,J=3.7Hz,1H),4.22(d,J=2.4Hz,1H),4.21–4.17(m,1H),4.09–4.05(m,2 H),3.96–3.90(m,2H),3.81(s,3H),3.45(s,1H),3.43(d,J=10.2Hz,1H),3. 15(ddd,J=11.9,7.4,4.9Hz,1H),3.06(d,J=18.1Hz,1H),2.93(dd,J=18.2, 9.0Hz,1H),2.86–2.79(m,1H),2.61(dt,J=15.0,4.7Hz,1H),2.56–2.48(m, 2H),2.37(s,3H),2.29–2.23(m,4H),2.22(s,3H),2.07(s,3H),1.26(s,2H).

[0088] Example 1.3:

[0089] (1'R,6R,6aR,7R,13S,14R,16R)-14-cyano-6'-(hydroxymethyl)-8-hydroxy-9-methoxy-4,10,23-trimethyl-19-oxo-2',3',4',6,7,9',12,13,14,16-decahydro-6aH-spiro[7,13-imino-6,16-(cyclothiopropoxymethylene)[1,3]dioxapentanoc[7,8]isoquinolino[3,2-b][3]benzozocin-20,1'-pyridino[3,4-b]indole]-5-ylacetate

[0090]

[0091] Implementation method:

[0092]

[0093] Step 1: Synthesis of methyl 3-(2-amino-2-oxoacetyl)-1H-indole-5-carboxylic acid

[0094]

[0095] Oxaloyl chloride (1.52 g, 11.99 mmol) was added dropwise to a tetrahydrofuran (100 mL) solution of methyl 1H-indole-5-carboxylate (1.3-a, 1.75 g, 9.99 mmol) at 0 °C. The mixture was stirred at room temperature for 2 hours. Stirring was stopped after complete conversion was monitored by TLC. The reaction solution was concentrated under reduced pressure and dissolved in tetrahydrofuran (50 mL). The solution was then cooled to 0 °C and a methanol solution of ammonia (8.0 mL, 7 N, 56.00 mmol) was added dropwise. The mixture was stirred at room temperature for 10 hours. The reaction solution was concentrated under reduced pressure to give crude methyl 3-(2-amino-2-oxoacetyl)-1H-indole-5-carboxylate as a pale yellow solid (1.3-b, 1.48 g, 60%).

[0096] LC-MS:(ES + ):m / z 247.1[M+H] + .

[0097] Step 2: Synthesis of compound (3-(2-aminoethyl)-1H-indol-5-yl)methanol

[0098]

[0099] Lithium aluminum hydride (2.28 g, 60.11 mmol) was added in portions to a tetrahydrofuran (30 mL) solution of intermediate methyl 3-(2-amino-2-oxoacetyl)-1H-indole-5-carboxylic acid (1.3-b, 1.48 g, 6.01 mmol). The reaction mixture was stirred until no gas was generated, then heated to reflux and stirred for 12 hours. After the reaction solution was cooled to room temperature, 15% sodium hydroxide aqueous solution (2.2 mL) and water (6.6 mL) were slowly added dropwise. After stirring for 2 hours, the mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was diluted with ethyl acetate (50 mL) and water (50 mL), and the organic phase was collected, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by preparative HPLC (mobile phase was 10%–95% acetonitrile aqueous solution, aqueous phase contained 0.1% (v / v) formic acid) to obtain compound (3-(2-aminoethyl)-1H-indol-5-yl)methanol as a pale yellow solid (1,3-c, 480 mg, 42%).

[0100] LC-MS:(ES + ):m / z 191.1[M+H] + .

[0101] Step 4: Synthesis of Compound 1.3: (1'R,6R,6aR,7R,13S,14R,16R)-14-cyano-6'-(hydroxymethyl)-8-hydroxy-9-methoxy-4,10,23-trimethyl-19-oxo-2',3',4',6,7,9',12,13,14,16-decahydro-6aH-spiro[7,13-imino-6,16-(cyclothiopropoxymethylene)[1,3]dioxapentanoc[7,8]isoquinolino[3,2-b][3]benzozocin-20,1'-pyridino[3,4-b]indole]-5-ylacetate

[0102]

[0103] Intermediate Ia (20 mg, 0.032 mmol) was added to a 2 mL acetic acid solution of intermediate (3-(2-aminoethyl)-1H-indol-5-yl)methanol (1.3-d, 18 mg, 0.096 mmol). The mixture was reacted at room temperature for 24 hours. Stirring was stopped after the reaction was completed by TLC monitoring. The reaction solution was concentrated under reduced pressure, diluted with acetonitrile (1 mL) and water (2 mL), and purified by preparative HPLC (mobile phase: 10%–95% aqueous acetonitrile solution, aqueous phase containing 0.1% (v / v) formic acid) to give compound 1.3 as a white solid (19 mg, 74%).

[0104] LC-MS:(ES + ):m / z 794.3[M+H] + .

[0105] 1H NMR (400MHz, CDCl3) δ7.79(s,1H),7.41(s,1H),7.25(s,1H),7.14(dd,J=8.4,1.7Hz,1H),6.68(s,1H),6.26(d,J=1.3Hz ,1H),6.06(d,J=1.4Hz,1H),5.81(s,1H),5.11(d,J=11.6Hz,1H),4.72(s,2H),4.58(s,1H),4.36(s,1H),4.30(dd,J=4.6 ,1.6Hz,1H),4.28–4.18(m,2H),3.84(s,3H),3.50–3.39(m,2H),3.13–3.09(m,1H),3.06(s,1H),2.98(d,J=9.0Hz,1H), 2.90–2.82(m,1H),2.69–2.63(m,1H),2.57(d,J=15.3Hz,2H),2.40(s,3H),2.35–2.26(m,4H),2.24(s,3H),2.09(s,3H).

[0106] Example 1.4:

[0107] (1'R,6R,6aR,7R,13S,14R,16R)-14-cyano-6'-((3-hydroxyazacyclobutane-1-yl)methyl)-8-hydroxy-9-methoxy-4,10,23-trimethyl-19-oxo-2',3',4',6,7,9',12,13,14,16-decahydro-6aH-spiro[7,13-imino-6,16-(cyclothiopropoxymethylene)[1,3]dioxapentan[7,8]isoquinolino[3,2-b][3]benzozocin-20,1'-pyridino[3,4-b]indole]-5-ylacetate

[0108]

[0109] Implementation method:

[0110]

[0111] Step 1: Synthesis of compound (2-(5-(hydroxymethyl)-1H-indol-3-yl)ethyl)tert-butyl carbamate

[0112]

[0113] Di-tert-butyl dicarbonate (803 mg, 3.68 mmol) was added dropwise to a solution of (3-(2-aminoethyl)-1H-indol-5-yl)methanol (1.3-c, 500 mg, 2.63 mmol) and triethylamine (532 mg, 5.26 mmol) in dichloromethane (10 mL) at room temperature. The mixture was stirred at room temperature for 12 hours. Stirring was stopped after complete conversion was monitored by TLC. The reaction solution was diluted with dichloromethane (15 mL) and water (15 mL), the organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluting with petroleum ether containing 40% ethyl acetate) to give the product (2-(5-(hydroxymethyl)-1H-indol-3-yl)ethyl)carbamate as a gray solid (1.4-a, 414 mg, 54%).

[0114] LC-MS:(ES + ):m / z 235.1[M-55] + .

[0115] Step 2: Synthesis of compound (2-(5-formyl-1H-indol-3-yl)ethyl)tert-butyl carbamate

[0116]

[0117] Dess-Martin oxidant (726 mg, 1.71 mmol) was added to a solution of (2-(5-(hydroxymethyl)-1H-indol-3-yl)ethyl)carbamate (tert-butyl) in dichloromethane (10 mL) at room temperature. The mixture was stirred for 12 hours at room temperature. Stirring was stopped after complete conversion was monitored by TLC. The reaction solution was diluted with dichloromethane (15 mL) and water (15 mL), the organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluting with petroleum ether containing 20% ​​ethyl acetate) to give the product (2-(5-formyl-1H-indol-3-yl)ethyl)carbamate as a white solid (1.4-b, 247 mg, 60%).

[0118] LC-MS:(ES + ):m / z 233.1[M-55] + .

[0119] Step 3: Synthesis of tert-butyl carbamate (2-(5-((3-hydroxyazacyclobutane-1-yl)methyl)-1H-indol-3-yl)ethyl)carbamate

[0120]

[0121] At room temperature, tert-butyl carbamate (1,4-b, 150 mg, 0.52 mmol) in 1,2-dichloroethane (10 mL) was mixed with aziridine-3-ol hydrochloride (1,4-c, 285 mg, 2.60 mmol) and triethylamine (362 μL, 2.60 mmol), followed by the addition of NaBH(OAc)3 (551 mg, 2.60 mmol). The mixture was stirred at room temperature for 36 hours. After the reaction was completely converted by TLC monitoring, stirring was stopped. The reaction solution was quenched with methanol (3 mL), concentrated under reduced pressure, diluted with acetonitrile (3 mL) and water (4 mL), and purified by preparative HPLC (mobile phase was 10%–95% acetonitrile aqueous solution, aqueous phase contained 0.1% (v / v) formic acid) to obtain compound (2-(5-(((3-hydroxyazacyclobutane-1-yl)methyl)-1H-indol-3-yl)ethyl)carbamate tert-butyl ester as a white solid (1,4-d, 100 mg, 56%).

[0122] LC-MS:(ES + ):m / z 346.1[M+H] + .

[0123] Step 4: Synthesis of compound 1-((3-(2-aminoethyl)-1H-indol-5-yl)methyl)azacyclobutane-3-ol trifluoroacetate

[0124]

[0125] Trifluoroacetic acid (1 mL) was added to dichloromethane (2 mL) containing the intermediate (2-(5-(((3-hydroxyazacyclobutane-1-yl)methyl)-1H-indol-3-yl)ethyl)carbamate (1,4-d, 100 mg, 0.60 mmol), and the mixture was reacted at room temperature for 0.5 h. Stirring was stopped after the reaction was completed as monitored by TLC. Water (5 mL) was added to the reaction mixture, and the mixture was washed with dichloromethane (5 mL × 2). The aqueous phase was purified by preparative HPLC (mobile phase: 5%–90% acetonitrile aqueous solution, aqueous phase containing 0.5% (v / v) trifluoroacetic acid) to obtain the product 1-((3-(2-aminoethyl)-1H-indol-5-yl)methyl)azacyclobutane-3-ol trifluoroacetate. The trifluoroacetate was a white solid (1,4-e, 60 mg, 61%).

[0126] LC-MS:(ES + ):m / z 246.1[M+H] + .

[0127] Step 5: Synthesis of compound 1.4: (1'R,6R,6aR,7R,13S,14R,16R)-14-cyano-6'-((3-hydroxyazacyclobutane-1-yl)methyl)-8-hydroxy-9-methoxy-4,10,23-trimethyl-19-oxo-2',3',4',6,7,9',12,13,14,16-decahydro-6aH-spiro[7,13-imino-6,16-(cyclothiopropoxymethylene)[1,3]dioxapentano[7,8]isoquinolino[3,2-b][3]benzozocin-20,1'-pyridino[3,4-b]indole]-5-yl acetate

[0128]

[0129] Intermediate Ia (20 mg, 0.032 mmol) was added to a white solid (14-e, 33 mg, 0.096 mmol) of intermediate 1-((3-(2-aminoethyl)-1H-indol-5-yl)methyl)azacyclobutane-3-ol trifluoroacetate. The mixture was reacted at room temperature for 24 hours. After the reaction was completed by TLC monitoring, stirring was stopped. The reaction solution was concentrated under reduced pressure, diluted with acetonitrile (1 mL) and water (2 mL), and purified by preparative HPLC (mobile phase: 10%–95% aqueous acetonitrile solution, aqueous phase containing 0.1% (v / v) formic acid) to obtain compound 1.4 as a white solid (21 mg, 77%).

[0130] LC-MS:(ES + ):m / z 849.3[M+H] + .

[0131] 1H NMR(400MHz, CDCl3)8.59(s,1H),7.92(s,1H),7.33(s,1H),7.07(d,J=8.3Hz,1H),6.65(s,1H),6.24(d,J=1.3Hz,1H),6.03(d,J=1.4Hz,1H),5 .08(d,J=11.6Hz,1H),4.57(s,1H),4.41(s,1H),4.33(s,1H),4.28(dd,J=4.6,1.5Hz,1H),4.19(dd,J=12.0,2.1Hz,2H),3.98(s,2H),3.86(q,J =9.5Hz,2H),3.80(s,3H),3.68(d,J=19.1Hz,2H),3.43(t,J=7.3Hz,2H),3.16(td,J=7.6,3.9Hz,1H),3.02(t,J=20.3Hz,2H),2.96–2.89(m,1H ),2.83(dt,J=10.6,5.1Hz,1H),2.63(dt,J=9.3,4.8Hz,2H),2.55(dd,J=14.4,7.1Hz,2H),2.36(s,3H),2.24(s,3H),2.22(s,3H),2.06(s,3H).

[0132] The following example in Table 1 was completed using the above synthesis method:

[0133] Table 1

[0134]

[0135]

[0136] Example 2.1

[0137] (1'R,6R,6aR,7R,13S,14S,16R)-6'-(3-hydroxyazacyclobutane-1-yl)-8,14-dihydroxy-9-methoxy-4,10,23-trimethyl-19-oxo-2',3',4',6,7,9',12,13,14,16-decahydro-6aH-spiro[7,13-imino-6,16-(cyclothiopropoxymethylene)[1,3]dioxapentano[7,8]isoquinolino[3,2-b][3]benzozocin-20,1'-pyridino[3,4-b]indole]-5-ylacetate

[0138]

[0139] Implementation method:

[0140]

[0141] step:

[0142] To a solution of compound 1.1 (12 mg, 0.014 mmol) in acetonitrile (1.3 mL), water (1 mL) and silver nitrate (49 mg, 0.29 mmol) were added, and the mixture was reacted at room temperature in the dark for 12 hours. The reaction was monitored by TLC, and stirring was stopped. The reaction solution was quenched with a mixed solution of saturated sodium carbonate and saturated sodium chloride (1:1, 4 mL), stirred at room temperature for 10 minutes, and then filtered to remove insoluble matter. The filtrate was extracted with dichloromethane (10 mL × 2). The organic phases were combined and concentrated under reduced pressure. The residue was diluted with acetonitrile (1 mL) and water (2 mL) and purified by preparative HPLC (mobile phase: 10%–95% aqueous acetonitrile solution; aqueous phase: 0.05% (v / v) trifluoroacetic acid) to give compound 2.1 as a white solid (8 mg, 67%).

[0143] LC-MS:(ES + ):m / z 808.3[M-17] + .

[0144] 1 H NMR (400MHz, CDCl3) δ7.53(s,1H),7.35(d,J=2.6Hz,1H),7.14–7.06(m,1H),6.66(s,1H),6.42(d,J=2.2Hz,1H),6.39(dd,J=8.5,2.3 Hz,1H),6.19(d,J=1.4Hz,1H),5.99(s,1H),5.74(s,1H),5.19(d,J=11.5Hz,1H),4.82(s,1H),4.73–4.66(m,1H),4.54–4.35(m,3H),4 .26–4.18(m,1H)4.17–4.08(m,2H),3.82(s,3H),3.59(dd,J=7.8,4.7Hz,2H),3.50(d,J=4.6Hz,1H),3.25–3.11(m,2H),3.06–2.96(m ,1H),2.92–2.77(m,2H),2.64–2.56(m,1H),2.51(d,J=11.6Hz,1H),2.46(s,1H),2.37(s,3H),2.26(s,3H),2.19(s,3H),2.05(s,3H).

[0145] The following two examples were completed using the above synthesis method:

[0146] Table 2

[0147]

[0148]

[0149]

[0150] Example 3.1 Antitumor bioactivity

[0151] In vitro detection of inhibitory activity against tumor cell proliferation:

[0152] Miacapa-2, BxPC-3, BT474, SW480, WiDr, OVCAR-3, SK-OV-3, and H-1048 tumor cells were cultured using standard methods at 37°C, in a 5% CO2 incubator, and in the specified cell culture medium. The tumor cells were digested with trypsin using standard methods, and the cells were collected and counted. The cells were resuspended in the corresponding detection medium, and 2000-5000 cells / well were added to 96-well plates at 80 μL / well. The test drugs were diluted with culture medium. For Dxd, rupettedine, and their derivatives, the concentrations started at 1000 nM and were diluted 3-fold in eight concentration gradients. For Dxd, the molecular concentration started at 5000 nM and was diluted 3-fold in eight concentration gradients. 20 μL of each of these solutions was added to 96-well plates containing 180 μL of culture medium and cells. The cells were incubated at 37℃ and 5% CO2 for 3 days. After incubation, 20 μL of CCK8 reagent was added to each well. After reacting for 2 hours, the absorbance at 450 nM was measured using a microplate reader. The experimental results are shown in Table 3.

[0153] Table 3. Inhibition rate (IC50) of the compounds of the present invention against various tumor cells. 50 (nM)

[0154]

[0155]

[0156] Test results show that the compounds of this invention have extremely strong cell-killing effects in a variety of tumor cells.

[0157] Example 3.2 Lymphocyte killing activity

[0158] In vitro assay for inhibitory activity against lymphocytes:

[0159] Step 1:

[0160] Preparation of RPMI 1640 medium: Add 10 mL FBS and 1 mL antibiotics to 89 mL of RPMI 1640 medium. Add 4 mL of the prepared RPMI 1640 medium (89 mL RPMI 1640 medium with 10 mL FBS) to a 15 mL centrifuge tube. Quickly transfer the PBMC cell cryovial from the liquid nitrogen tube to a 37°C water bath and thaw the cells. Transfer the cells from the cryovial to a 15 mL test tube. Rinse the cryovial with 1 mL of cell culture medium and transfer it to the 15 mL test tube. Add cell culture medium to a final volume of 10 mL. Centrifuge at 2000 RPM for 7 minutes, discard the supernatant, and resuspend the cells in 1 mL of cell culture medium, transferring slowly up and down. Add cell culture medium to a final volume of 10 mL, centrifuge at 2000 RPM for 7 minutes. Discard the supernatant and resuspend the cells in 4 mL of cell culture medium. Count the cells and determine cell viability using trypan blue staining, adjusting the PBMC cell density to 1 × 10⁻⁶. 6 / mL cell culture flasks were placed in a carbon dioxide incubator and incubated overnight.

[0161] Step Two:

[0162] On day 2, the small molecule drug was diluted to the target concentration using cell culture medium. Overnight PBMC cells were used, and cell viability was determined by trypan blue staining. The cell density was adjusted to 2.5 × 10⁻⁶. 6 At a concentration of 1 / mL, 180 μL of cells were seeded per well in a 96-well plate. 20 μL of the prepared compound was added. An equal volume of DMSO was added to the blank control. After incubation in a CO2 incubator for 28 hours, the cells were centrifuged, the supernatant discarded, and the cells were resuspended in 200 μL of FACS buffer (5% FBS in PBS). The cells were washed twice. 2.5 μg of Fc blocking agent was added to each sample. After incubation at room temperature for 10 minutes, 10 μL of flow cytometry antibody and 87.5 μL of flow cytometry buffer were added, and the cells were incubated at 4°C for 1 hour. The cells were washed twice with 200 μL of FACS buffer (centrifuged at 500g for 7 minutes). The cells were resuspended in 200 μL of FACS buffer and analyzed by flow cytometry. 50,000 samples were collected.

[0163] Data calculation method: (1 - number of lymphocytes in the experimental group / number of lymphocytes in the blank control group) × 100%, and the results are shown in Table 4.

[0164] Table 4. Killing effect of the compounds of the present invention on human peripheral blood lymphocytes (%)

[0165] Compound (concentration) B-cell killing effect T-cell killing effect Blank control (DMSO) 0 0 Dxd(30nM) 0 4 Rupettedine (30 nM) 82 54 Compound 1.2 (30 nM) 48 20 Compound 1.2 (100 nM) 73 37 Compound 2.2 (30 nM) 87 76 Compound 2.2 (100 nM) 90 84 Compound 1.3 (30 nM) 4 ND Compound 1.3 (100 nM) 43 ND Compound 2.3 (30 nM) 52 ND

[0166] “ND” indicates that it was not detected.

[0167] Test results show that the compound described in this invention has a strong cytotoxic effect on lymphocytes, and this cytotoxic effect on lymphocytes gives it the potential to treat autoimmune diseases.

[0168] The applicant declares that this invention illustrates the sea sucrose derivatives and their applications through the above embodiments, but this invention is not limited to the above embodiments, that is, it does not mean that this invention must rely on the above embodiments to be implemented. Those skilled in the art should understand that any improvements to this invention, equivalent substitutions of the raw materials of the product, addition of auxiliary components, and selection of specific methods, etc., all fall within the protection scope and disclosure scope of this invention.

Claims

1. A tunicamycin derivative or a tautomer, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, characterized by, The sea sponge derivative has a structure as shown in general formula (I): wherein: R1is selected from cyano, hydroxy, halogen, carboxyl, R2is selected from the following groups: R3is selected from the group consisting of hydroxy, carboxy, mercapto or 2-mercaptoacetylamino, R4, R5, R6, R7and R8are independently selected from the group consisting of hydrogen, C1-C5alkyl or hydroxy substituted C1-C5alkyl; the wavy line represents the point of attachment of the group; m and n are each independently selected from 1, 2, 3 or 4.

2. The tunicamycin derivative according to claim 1, or a tautomer, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, characterized in that, R2is selected from the following groups: wherein R3 is selected from the group consisting of hydroxy, carboxy, mercapto or 2-mercaptoacetylamino, R4, R7 and R8 are independently selected from the group consisting of hydrogen, C1-C5 alkyl or hydroxy substituted C1-C5 alkyl, and the wavy line represents the point of attachment of the group.

3. The tunicamycin derivative according to claim 1 or 2, or a tautomer, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, characterized in that, R3is selected from hydroxyl, amino, carboxyl, methylamino, ethylamino, hydroxyethylamino, mercapto or 2-mercaptoacetylamino.

4. The tunicamycin derivative according to claim 1 or 2, or a tautomer, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, characterized in that, R4is selected from methyl, ethyl, isopropyl or hydroxyethyl.

5. The tunicamycin derivative according to any one of claims 1 to 4, or a tautomer, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, characterized in that, R2is selected from the following groups: The wavy line represents the linking site of the group.

6. The tunicamycin derivative according to any one of claims 1 to 5, or a tautomer, an entiomer thereof, or a pharmaceutically acceptable salt thereof, characterized in that, The sea sponge derivative is selected from any one of the following compounds:

7. A pharmaceutical composition, characterized by, The pharmaceutical composition comprises at least one sea sponge derivative as claimed in any one of claims 1-6, or a tautomer, stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

8. An antibody-drug conjugate, characterized in that, The small molecule drug moiety of the antibody-drug conjugate comprises a sea sponge derivative as claimed in any one of claims 1-6, or a tautomer, stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

9. Use of a sea sponge derivative as claimed in any one of claims 1-6, or a tautomer, stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as claimed in claim 7, in the preparation of a medicament for treating and / or preventing an autoimmune disease. Preferably, the autoimmune disease comprises psoriasis, psoriatic arthritis, systemic lupus erythematosus, lupus nephritis, rheumatoid arthritis, multiple sclerosis, type I diabetes, anti-glomerular basement membrane antibody nephritis, Crohn's disease, ulcerative colitis, myasthenia gravis or vitiligo.

10. Use of a sea sponge derivative as claimed in any one of claims 1-6, or a tautomer, stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as claimed in claim 7, in the preparation of a medicament for treating and / or preventing a tumor. Preferably, the tumor comprises non-small cell lung cancer, small cell lung cancer, colorectal cancer, breast cancer, pancreatic cancer, sarcoma, ovarian cancer, prostate cancer, gastric cancer, liver cancer, kidney cancer or hematological tumor.

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