A BBI608 derivative, its preparation method, and its application in the preparation of drugs for treating STAT3-mediated diseases.

Abstract

This invention provides a BBI608 derivative, its preparation method, and its application in the preparation of drugs for treating STAT3-mediated diseases, belonging to the field of pharmaceutical synthesis technology. The BBI608 derivative provided by this invention has the structure shown in Formula I, II, or III; the BBI608 derivative provided by this invention exhibits high inhibitory activity against HepG2 cells and can be used as a drug for treating STAT3-mediated diseases. The results of the examples show that the BBI608 derivative provided by this invention has inhibitory activity against HepG2 IC50 cells. 50 The values ​​ranged from 0.08 nM to 4.81 μM, exhibiting excellent inhibitory activity. Experimental results showed that SC-7 exhibited the strongest inhibitory activity against HepG2, with an IC50 value of [missing value]. 50 It is 0.08 μM.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical synthesis technology, and in particular to a BBI608 derivative, its preparation method, and its application in the preparation of drugs for treating STAT3-mediated diseases. Background Technology

[0002] Cancer, medically termed malignant tumor and in traditional Chinese medicine as "rock," is a disease caused by the abnormality of the mechanisms controlling cell growth and proliferation. Besides uncontrolled proliferation, cancer cells can also locally invade normal tissues and even metastasize to other parts of the body via the circulatory system. Cancer stem cells refer to cancer cells with stem cell properties, meaning they possess the ability to self-replicate and differentiate into multiple cells. These cells are generally considered to have the potential to develop into cancer, especially as they become a source of new cancers after metastasis. If cancer stem cells are not completely eliminated during cancer treatment, cancer is prone to recurrence and metastasis. Current research has identified several signaling pathways involved in tumor stem cells, including Wnt, Hedgehog (Hh), Notch, BMP, Bmi-1, PI3K / Akt, and STAT3. These pathways regulate the self-renewal and differentiation of tumor stem cells. The first three signaling pathways targeting tumor stem cells are the most thoroughly studied, while STAT3 is currently the most researched and is the main target pathway for tumor stem cell therapy. These signaling pathways have opened up a new field for tumor research and brought new hope for tumor treatment (Journal of Cell Science, 2005, 118(Sup 4): 665-672.).

[0003] The gene encoding STAT3 in the human body is located on chromosome 17 (q21.1–q21.2). This gene contains 24 exons, with a full-length DNA of 4815 bp, totaling 770 amino acids and a molecular weight of approximately 92,000 Da. Other STAT proteins have similar structures, mainly including: 1. an N-terminal conserved amino acid domain, involved in STAT3 tetramerization; 2. a 4-helix bundle (amino acid residues 138–320), the site of action for transcription factors and regulatory proteins; and 3. a DNA-specific binding domain (amino acid residues 321–465), which specifically binds to active IFN-γ palindromic sequence elements. 4. Linking region (amino acid residues 466-585), a stable DNA-binding domain; 5. SH2 domain (amino acid residues 586-688), a small molecule inhibitor binding site for STAT3; 6. Phosphorylation site (amino acid residues 702-716), phosphorylation triggers dimerization, which in turn forms a dimer in the cytoplasm; 7. C-terminal transcription activation domain, where phosphorylation of tryptophan (Ser727) or tyrosine near the C-terminus (Try705) activates STAT3 (Nature, 1998, 394(6689): 145-151.). In addition to extracellular signal stimulation, the signal transduction of STAT3 protein can be activated by receptor or non-receptor tyrosine kinases through a tyrosine phosphorylation cascade. So far, signaling factors that can activate STAT3 protein have been found to include epidermal growth factor receptor (EGFR), vascular epidermal growth factor receptor (VEGFR), fibroblast growth factor (FGFR), Src family-associated kinase, Abl family-associated kinase, insulin-like growth factor receptor (IGFRs), hepatocyte growth factor receptor (HGFRs), interleukin-6 (IL-6), etc. (American Journal of Pathology, 2004, 165(5):1449-1460.) After STAT3 is stimulated by extracellular signals, tyrosine is phosphorylated and STAT3 monomers are activated. The activated STAT3 monomers interact with the SH2 domain to form homodimers, which enter the cell nucleus and bind to specific DNA sequences, thereby regulating the transcription and expression of downstream target genes. After STAT3 completes signal transduction, nucleoprotein tyrosine dephosphorylase dephosphorylates phosphorylated STAT3 and returns it to the cytoplasm (Journal of Clinical Investigation, 2002, 109(9): 1139-1142.).

[0004] Under normal circumstances, STAT3 exists in the cytoplasm in an inactive form. Recent studies have demonstrated that STAT3 signaling regulates the self-renewal, differentiation, and apoptosis of tumor stem cells. STAT3 plays a crucial role in maintaining the self-renewal of tumor stem cells; therefore, STAT3 is also a novel target for targeted therapy of tumor stem cells. Summary of the Invention

[0005] The purpose of this invention is to provide a BBI608 derivative, its preparation method, and its application in the preparation of drugs for treating STAT3-mediated diseases. The BBI608 derivative provided by this invention exhibits high inhibitory activity against HepG2 cells and can be used as a drug for treating STAT3-mediated diseases.

[0006] To achieve the above-mentioned objectives, the present invention provides the following technical solution:

[0007] This invention provides a BBI608 derivative, characterized in that it has the structure shown in Formula I, II, or III:

[0008]

[0009] In formulas I, II, and III: X is an O, N, or S atom;

[0010] R1 is independently one of hydroxyl, substituted or unsubstituted C1-C10 alkylamine, substituted or unsubstituted C1-C10 alkylolamine, substituted or unsubstituted C3-C7 cycloalkylamine, substituted or unsubstituted 5-10 membered heterocyclic amine or heteroaromatic amine;

[0011] R2 is independently one of C1-C6 alkyl or C1-C6 alkoxy.

[0012] Preferably, the substituted or unsubstituted 5- to 10-membered heterocyclic amine or heteroaromatic amine contains 1 to 3 heteroatoms, wherein the heteroatoms are N, O, or S.

[0013] Preferably, R1 is independently hydroxyl, substituted or unsubstituted methylamino, substituted or unsubstituted diethylamino, D-valine methyl ester, substituted or unsubstituted isopropylamino, substituted or unsubstituted isopropanolamino, substituted or unsubstituted diethanolamino, substituted or unsubstituted isobutylamino, substituted or unsubstituted cyclopentanamino, substituted or unsubstituted cyclohexylamino, substituted or unsubstituted piperidinyl, substituted or unsubstituted piperazine, substituted or unsubstituted morpholino, substituted or unsubstituted pyrazine, substituted or unsubstituted piperazine, substituted or unsubstituted tetrahydropyrrole, substituted or unsubstituted tetrahydrofuran, or substituted or unsubstituted furanyl.

[0014] Preferably, R1 is independently -OH, -N(CH3)2, -N(CH2CH3)2, -NCHOHCH3, -N(CH2CH2OH)2, N-BOC ethylenediamine, D-proline methyl ester, D-valine methyl ester, tetrahydropyrrolyl, 2-(pyrrolidine-1-yl)ethane-1-amino, 2-(piperidin-1-yl)ethane-1-amino, 2-(morpholino-1-yl)ethane-1-amino, piperidinyl, (S)-piperidin-3-carboxylic acid ethyl ester, (R)-piperidin-3-carboxylic acid ethyl ester, (S)-piperidin-3-ylcarbamate tert-butyl ester. One of (R)-piperidin-3-ylcarbamate tert-butyl ester, 4-aminopiperidin-1-carbamate tert-butyl ester, piperidin-4-carboxylic acid ethyl ester, piperidin-4-carboxylic acid methyl ester, 1-(bis(4-fluorophenyl)methyl)piperazinyl, 1-(bis(phenyl)methyl)piperazinyl, piperazin-1-carboxylic acid tert-butyl ester, 1-benzylpiperazinyl, 1-methylpiperazinyl, 2-(piperazin-1-yl)ethane-1-olyl, 1-(piperazin-1-yl)ethane-1-oneyl, (1-(2,2-diphenylacetyl)piperazin-2-carbonyl)-L-valine methyl ester, diphenylamino, and dibenzylamino.

[0015] Preferably, R2 is independently one of C1-C4 alkyl or C1-C4 alkoxy.

[0016] Preferably, it has any one of the structures shown in SG-1 to SG-27:

[0017]

[0018]

[0019] This invention provides a method for preparing the above-mentioned BBI608 derivatives, comprising the following steps:

[0020] (i) When the BBI608 derivative has the structure shown in Formula I, the preparation method includes the following steps:

[0021] Compound a undergoes a first nucleophilic substitution reaction with a nitrating agent to give compound b;

[0022] Compound b and compound 1 undergo a first cladding reaction to yield compound c;

[0023] Compound c undergoes a second nucleophilic substitution reaction with a nitrating agent to give compound d;

[0024] The compound d undergoes a first reduction reaction to yield compound e;

[0025] Compound e and compound 2 undergo a second ring-cladding reaction to yield compound f;

[0026] The compound f undergoes a first hydrolysis reaction to obtain compound g;

[0027] The compound g undergoes a first oxidation reaction to yield compound h;

[0028] When R1 is a hydroxyl group, compound h is a BBI608 derivative having the structure shown in Formula I;

[0029] When R1 is one of the following: substituted or unsubstituted C1-C10 alkylamine, substituted or unsubstituted C1-C10 alkylolamine, substituted or unsubstituted C3-C7 cycloalkylamine, substituted or unsubstituted 5-10 membered heterocyclic amine or heteroaromatic amine, in the presence of 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate and N,N-diisopropylethylamine, compound h undergoes a first acylation reaction with compound 3 to obtain a BBI608 derivative having the structure shown in Formula I;

[0030] Compound 1; PhNOS compound 2; R1H compound 3;

[0031]

[0032] (ii) When the BBI608 derivative has the structure shown in Formula II, the preparation method includes the following steps:

[0033] Compound g and compound 4 undergo a second acylation reaction to yield compound i;

[0034] Compound i undergoes a first electrophilic substitution reaction with a Grignard reagent to obtain compound j;

[0035] Compound j undergoes a second reduction reaction to yield compound k;

[0036] The compound k undergoes a second oxidation reaction to yield compound l;

[0037] When R1 is a hydroxyl group, compound 1 is a BBI608 derivative having the structure shown in Formula II;

[0038] When R1 is one of the following: a substituted or unsubstituted C1-C10 alkylamine group, a substituted or unsubstituted C1-C10 alkylolamine group, a substituted or unsubstituted C3-C7 cycloalkylamine group, a substituted or unsubstituted 5-10 membered heterocyclic amine group, or a heteroaromatic amine group, the compound l undergoes a third nucleophilic substitution reaction with a chlorinating reagent to give compound m.

[0039] The compound m undergoes a fourth nucleophilic substitution reaction with the compound 3 to obtain a BBI608 derivative having the structure shown in Formula II.

[0040] Compound 4; R1H compound 3;

[0041]

[0042] (iii) When the BBI608 derivative has the structure shown in Formula III, the preparation method includes the following steps:

[0043] Compound e and compound 5 undergo a third ring-cladding reaction to yield compound n;

[0044] The compound n undergoes a second hydrolysis reaction to yield compound o;

[0045] Compound o undergoes a third oxidation reaction to yield compound p;

[0046] When R1 is a hydroxyl group, compound p is a BBI608-type derivative having the structure shown in Formula III;

[0047] When R1 is one of the following: substituted or unsubstituted C1-C10 alkylamine, substituted or unsubstituted C1-C10 alkylamino, substituted or unsubstituted C3-C7 cycloalkylamine, substituted or unsubstituted 5-10 membered heterocyclic amine or heteroaromatic amine, in the presence of 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate and N,N-diisopropylethylamine, compound p and compound 3 undergo a third acylation reaction to obtain BBI608 derivatives having the structure shown in Formula III;

[0048] Compound 5; R1H compound 3;

[0049]

[0050] This invention provides the application of the above-mentioned BBI608 derivatives in the preparation of drugs for treating STAT3-mediated diseases.

[0051] Preferably, the STAT3-mediated diseases include one or more of colorectal cancer, lung cancer, melanoma, breast cancer, prostate cancer, kidney cancer, ovarian cancer, liver cancer, pancreatic cancer, multiple myeloma, and leukemia.

[0052] This invention provides a medicament for treating STAT3-mediated diseases, the active ingredient of which includes the above-mentioned BBI608 derivatives and their pharmaceutically acceptable salts.

[0053] This invention provides a BBI608-like derivative. Because the benzene ring of BBI608 is replaced with 1,2,5-thiadiazole and pyrazine, additional hydrogen bonding interactions can be formed with STAT3 proteins, resulting in most compounds exhibiting better activity than BBI608. The results of the examples show that the BBI608-like derivatives provided by this invention exhibit better activity against HepG2 IC50 proteins. 50 The concentration ranged from 0.08 μM to 4.81 μM, exhibiting strong inhibitory activity. Experimental results showed that SC-7 exhibited the strongest inhibitory activity against HepG2, with an IC50 value of [missing value]. 50 It is 0.08 μM. Attached Figure Description

[0054] Figure 1 The reaction route for BBI608 derivatives having the structure shown in Formula I;

[0055] Figure 2 The reaction route for BBI608 derivatives with the structure shown in Formula II;

[0056] Figure 3 The reaction route is for BBI608 derivatives with the structure shown in Formula III. Detailed Implementation

[0057] This invention provides a BBI608 derivative, characterized in that it has the structure shown in Formula I, II, or III:

[0058]

[0059] In formulas I, II, and III: X is an O, N, or S atom;

[0060] R1 is independently one of hydroxyl, substituted or unsubstituted C1-C10 alkylamine, substituted or unsubstituted C1-C10 alkylolamine, substituted or unsubstituted C3-C7 cycloalkylamine, substituted or unsubstituted 5-10 membered heterocyclic amine or heteroaromatic amine;

[0061] R2 is independently one of C1-C6 alkyl or C1-C6 alkoxy.

[0062] In this invention, R1 is independently one of a hydroxyl group, a substituted or unsubstituted C1-C10 alkylamine group, a substituted or unsubstituted C1-C10 alkylolamine group, a substituted or unsubstituted C3-C7 cycloalkylamine group, a substituted or unsubstituted 5-10 membered heterocyclic amine group, or a heteroaromatic amine group. In this invention, the C1-C10 alkylamine group is preferably one of methylamino, ethylenediamine, isopropylamino, isobutylamino, propylamino, butylamino, dimethylamino, ethylamino, or di-n-butylamine; the C1-C10 alkylolamine group is preferably one of isopropanolamine, diethanolamine, propanolamine, or ethanolamine; the C3-C7 cycloalkylamine group is preferably cyclopentanolamine or cyclohexylamino; in this invention, the substituted or unsubstituted 5-10 membered heterocyclic amine group or heteroaromatic amine group contains 1-3 heteroatoms, and the heteroatoms are N, O, or S; in this invention, the heteroatoms are preferably S.

[0063] In this invention, R1 is independently preferably hydroxyl, substituted or unsubstituted methylamino, substituted or unsubstituted diethylamino, D-valine methyl ester, substituted or unsubstituted isopropylamino, substituted or unsubstituted isopropanolamino, substituted or unsubstituted diethanolamino, substituted or unsubstituted isobutylamino, substituted or unsubstituted cyclopentanamino, substituted or unsubstituted cyclohexylamino, substituted or unsubstituted piperidinyl, substituted or unsubstituted piperazine, substituted or unsubstituted morpholino, substituted or unsubstituted pyrazine, substituted or unsubstituted piperazine, substituted or unsubstituted tetrahydropyrrole, substituted or unsubstituted tetrahydrofuran, or substituted or unsubstituted furanyl.

[0064] In this invention, R1 is independently preferably -OH, -N(CH3)2, -N(CH2CH3)2, -NCHOHCH3, -N(CH2CH2OH)2, N-BOC ethylenediamine, D-proline methyl ester, D-valine methyl ester, tetrahydropyrrolyl, 2-(pyrrolidine-1-yl)ethane-1-amino, 2-(piperidin-1-yl)ethane-1-amino, 2-(morpholino-1-yl)ethane-1-amino, piperidinyl, (S)-piperidin-3-carboxylic acid ethyl ester, (R)-piperidin-3-carboxylic acid ethyl ester, (S)-piperidin-3-ylcarbamate tert-butyl. One of the following: ester group, (R)-piperidin-3-ylcarbamate tert-butyl ester group, 4-aminopiperidin-1-carbamate tert-butyl ester group, piperidin-4-carboxylic acid ethyl ester group, piperidin-4-carboxylic acid methyl ester group, 1-(bis(4-fluorophenyl)methyl)piperazinyl, 1-(bis(phenyl)methyl)piperazinyl, piperazin-1-carboxylic acid tert-butyl ester group, 1-benzylpiperazinyl, 1-methylpiperazinyl, 2-(piperazin-1-yl)ethane-1-olyl, 1-(piperazin-1-yl)ethane-1-oneyl, (1-(2,2-diphenylacetyl)piperazin-2-carbonyl)-L-valine methyl ester group, diphenylamino, and dibenzylamino.

[0065] In this invention, R2 is independently one of C1-C4 alkyl or C1-C4 alkoxy; more preferably, it is methyl.

[0066] In this invention, the BBI608 derivatives have any one of the structures shown in SG-1 to SG-27:

[0067]

[0068] In this invention, the Chinese names of the structures shown in SG-1 to SG-27 are as follows:

[0069] N,N-Diethyl-5,9-dioxo-5,9-dihydrothiophene[2,3-g]quinoxaline-7-carboxamide (SG-1), (S)-1-(5,9-dioxo-5,9-dihydrothiophene[2,3-g]quinoxaline-7-carbonyl)piperidine-3-carboxylic acid ethyl ester (SG-2), (R)-1-(5,9-dioxo-5,9-dihydrothiophene[2,3-g]quinoxaline-7-carbonyl)piperidine-3-carboxylic acid ethyl ester (SG-3), 7-(piperidine-1-carbonyl)thiophene[2,3-g]quinoxaline-5,9-dione (SG-4), (2-(5,9-dioxo-5,9-dihydrothiophene[2,3-g]quinoxaline-7-carboxamido)ethyl)tert-butyl carbamate (S) G-5), 5,9-dioxo-N-(2-(piperidin-1-yl)ethyl)-5,9-dihydrothieno[2,3-g]quinoxaline-7-carboxamide (SG-6), 5,9-dioxo-N-(2-(pyrrolidone-1-yl)ethyl)-5,9-dihydrothieno[2,3-g]quinoxaline-7-carboxamide (SG-7), N-(2-morpholinoethyl)-5,9-dioxo-5,9-dihydrothieno[2,3-g]quinoxaline-7-carboxamide (SG-8), (S)-(1-(5,9-dioxy-5,9-dihydrothieno[2,3-g]quinoxaline-7-carbonyl)piperidin-3-yl)tert-butyl carbamate (SG-12), (R)-(1-(5,9-di... 5,9-dihydrothieno[2,3-g]quinoxaline-7-carbonyl)piperidin-3-yl)tert-butyl carbamate (SG-13), 4-(5,9-dioxo-5,9-dihydrothieno[2,3-g]quinoxaline-7-carboxamido)piperidin-1-carboxylic acid tert-butyl ester (SG-14), 7-(4-(bis(4-fluorophenyl)methyl)piperazine-1-carbonyl)thieno[2,3-g]quinoxaline-5,9-dione (SG-15), 7-(4-benzylpiperazine-1-carbonyl)thieno[2,3-g]quinoxaline-5,9-dione (SG-16), 4-(5,9-dioxo-5,9-dihydrothieno[2,3-g]quinoxaline-7-carbonyl)piperazine-1-carboxylic acid tert-butyl Ester (SG-18), 7-(4-benzylpiperazine-1-carbonyl)thieno[2,3-g]quinoxaline-5,9-dione (SG-19), 7-(4-methylpiperazine-1-carbonyl)thieno[2,3-g]quinoxaline-5,9-dione (SG-22), N-benzyl-5,9-dioxo-5,9-dihydrothieno[2,3-g]quinoxaline-7-carboxamide (SG-23), N,N-dibenzyl-5,9-dioxo-5-9-dihydrothieno[2,3-g]quinoxaline-7-carboxamide (SG-24), 6-(1-(4-methylpiperazine-1-yl)ethyl)thieno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-4,8-Diketone (SD-1), 6-(1-(4-(2-hydroxyethyl)piperazin-1-yl)ethyl)thieno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-4,8-dione (SD-2), 6-(1-(4-(pyrimidin-2-yl)piperazin-1-yl)ethyl)thieno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-4,8-dione (SD-3), (3S)-1-(1-(4,8-dioxo-4,8-dihydrothieno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-6-yl)ethyl)piperidine-3-carboxylic acid ethyl ester (SD-4), 1-(1-( 4,8-Dioxo-4,8-dihydrothieno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-6-yl)ethyl)piperidine-4-carboxylic acid ethyl ester (SD-5), 6-(1-(bis(2-hydroxyethyl)amino)ethyl)thieno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-4,8-dione (SD-6), 6-(4-methylpiperazine-1-carbonyl)thieno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-4,8-dione (SC-1), N,N-dimethyl-4,8-dioxo-4,8-dihydrothieno[2',3':4,5]benzo[1,2-c] [1,2,5]Thiadiazole-6-carboxamide (SC-2), 6-(pyrrolidine-1-carbonyl)thieno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-4,8-dione (SC-3), (R)-1-(4,8-dioxo-4,8-dihydrothiadiazole[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-6-carbonyl)piperidine-3-carboxylic acid ethyl ester (SC-4), 6-(4-acetylpiperazine-1-carbonyl)thieno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-4,8-dione (SC-5), (S)-1-(4,8-dioxo-4,8-dihydrothiadiazole) Benzo[1,2-c][1,2,5]thiadiazole-6-carbonyl)piperidine-3-carboxylic acid ethyl ester (SC-6), 4,8-dioxo-N-(2-(piperidin-1-yl)ethyl)-4,8-dihydrothiophene[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-6-carboxamide (SC-7), N-(2-hydroxypropyl)-4,8-dioxo-4,8-dihydrothiophene[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-6-carboxamide (SC-8), methyl(4,8-dioxo-4,8-dihydrothiophene[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-6-carboxamide (SC-8), methyl(4,8-dioxo-4,8-dihydrothiophene[2',3':4,5]benzo[1,2-c][1,2,5] Thiadiazole-6-carbonyl)-D-valine (SC-9), 6-(piperidin-1-carbonyl)thieno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-4,8-dione (SC-10), methyl(4,8-dioxo-4,8-dihydrothieno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-6-carbonyl)-D-proline (SC-11), N,N-diethyl-4,8-dioxo-4,8-dihydrothieno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-6-carboxamide (SC-13), 6- (4-(bis(4-fluorophenyl)methyl)piperazine-1-carbonyl)thieno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-4,8-dione (SC-14), 4-(4,8-dioxo-4,8-dihydrothieno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-6-carboxamido)piperidine-1-carboxylic acid tert-butyl ester (SC-15), 6-(4-diphenylmethylpiperazine-1-carbonyl)thieno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-4,8-dione (SC-16), 4-(4,8-dioxo-4, 8-Dihydrothiopheno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-6-carbonyl)piperazine-1-carboxylic acid tert-butyl ester (SC-17), 6-(4-benzylpiperidine-1-carbonyl)thiopheno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-4,8-dione (SC-22), 1-(4,8-dioxo-4,8-dihydrothiopheno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-6-carbonyl)piperidine-4-carboxylic acid ethyl ester (SC-23), (S)-(1-(4,8-dioxo-4,8-dihydrothiopheno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-6-carbonyl)piperidine-4-carboxylic acid ethyl ester (SC-23), (S)-(1-(4,8-dioxo-4,8-dihydrothiopheno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-6-carbonyl)piperidine-4-carboxylic acid tert-butyl ester [3':4,5]Benzo[1,2-c][1,2,5]thiadiazole-6-carbonyl)piperidin-3-yl)tert-butyl carbamate (SC-24), methyl 1-(4,8-dioxo-4,8-dihydrothiopheno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-6-carbonyl)piperidin-4-carboxylic acid (SC-25), (4-(4,8-dioxo-4,8-dihydrothiapheno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-6-carbonyl)-1-(2,2-diphenylacetyl)piperazine-2-carbonyl)-L-valine methyl ester (SC-27).

[0070] This invention provides a method for preparing the above-mentioned BBI608 derivatives, comprising the following steps:

[0071] (i) When the BBI608 derivative has the structure shown in Formula I, the preparation method includes the following steps:

[0072] Compound a undergoes a first nucleophilic substitution reaction with a nitrating agent to give compound b;

[0073] Compound b and compound 1 undergo a first cladding reaction to yield compound c;

[0074] Compound c undergoes a second nucleophilic substitution reaction with a nitrating agent to give compound d;

[0075] The compound d undergoes a first reduction reaction to yield compound e;

[0076] Compound e and compound 2 undergo a second ring-cladding reaction to yield compound f;

[0077] The compound f undergoes a first hydrolysis reaction to obtain compound g;

[0078] The compound g undergoes a first oxidation reaction to yield compound h;

[0079] When R1 is a hydroxyl group, compound h is a BBI608 derivative having the structure shown in Formula I;

[0080] When R1 is one of the following: substituted or unsubstituted C1-C10 alkylamine, substituted or unsubstituted C1-C10 alkylolamine, substituted or unsubstituted C3-C7 cycloalkylamine, substituted or unsubstituted 5-10 membered heterocyclic amine or heteroaromatic amine, in the presence of 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate and N,N-diisopropylethylamine, compound h undergoes a first acylation reaction with compound 3 to obtain a BBI608 derivative having the structure shown in Formula I;

[0081] Compound 1; PhNOS compound 2; R1H compound 3;

[0082]

[0083] In this invention, compound a undergoes a first nucleophilic substitution reaction with a nitrating reagent to obtain compound b. In this invention, the nitrating reagent preferably includes concentrated nitric acid, KNO3 / H2SO4, HNO3 / Ac2O, or ammonium nitrate / trifluoroacetic anhydride, more preferably concentrated nitric acid. In this invention, the molar ratio of compound a to the nitrating reagent is preferably 1:(3-10), more preferably 1:5. In this invention, the temperature of the first nucleophilic substitution reaction is preferably -10 to 20°C, more preferably 0°C, and the time is preferably 2 to 4 hours, more preferably 3 hours. After the first nucleophilic substitution reaction, in this invention, the resulting reaction solution is preferably poured into water, and after the yellow solid precipitates, it is filtered to obtain compound b.

[0084] After obtaining compound b, the present invention performs a first ring-closing reaction on compound b and compound 1 to obtain compound c. In the present invention, the first ring-closing reaction is preferably carried out under an alkaline environment. In the present invention, the base used for the first ring-closing reaction preferably includes triethylamine, pyridine, or N,N-diisopropylethylamine, potassium carbonate, cesium carbonate, potassium phosphate, and more preferably potassium carbonate; in the present invention, the molar ratio of compound b, compound 1, and base is preferably 1:(1.1-2):(1.1-3); more preferably 1:1.5:1.5. In the present invention, the first ring-closing reaction is preferably carried out in the presence of DMF. In the present invention, the temperature of the first ring-closing reaction is preferably 50-80°C, more preferably 70°C, and the time is preferably 6-12 h, more preferably 8 h. After the first ring-closing reaction, the present invention preferably pours the obtained reaction solution into water, precipitates a yellow solid, and then filters it to obtain compound c.

[0085] In this invention, compound c undergoes a second nucleophilic substitution reaction with a nitrating reagent to obtain compound d. Preferably, the nitrating reagent comprises one or more of concentrated nitric acid, KNO3 / H2SO4, HNO3 / Ac2O, and ammonium nitrate / trifluoroacetic anhydride, more preferably concentrated nitric acid. The molar ratio of compound c to the nitrating reagent is preferably 1:(3-10), more preferably 1:6. The temperature of the second nucleophilic substitution reaction is preferably -10 to 20°C, more preferably 0°C, and the time is preferably 2 to 4 hours, more preferably 3 hours. After the second nucleophilic substitution reaction, the resulting reaction solution is preferably poured into water, and after the yellow solid precipitates, it is filtered to obtain compound d.

[0086] In this invention, compound d undergoes a first reduction reaction to obtain compound e. In this invention, the reducing agent used in the first reduction reaction is preferably hydrogen, and the catalyst is preferably Pd-C. In this invention, the molar ratio of compound d to the catalyst is preferably 1:(0.1-0.2), more preferably 1:0.1. In this invention, the first reduction reaction is preferably carried out in the presence of anhydrous ethanol. In this invention, the temperature of the first reduction reaction is preferably 25-70°C, more preferably 60°C, and the time is preferably 10-15 h, more preferably 12 h. After the first reduction reaction, in this invention, the reaction solution is preferably cooled to room temperature, filtered, and the filtrate is concentrated under reduced pressure to obtain compound e.

[0087] In this invention, compound e and compound 2 undergo a second ring-closing reaction to obtain compound f. Preferably, the second ring-closing reaction is carried out in an alkaline environment. The base used in the second ring-closing reaction preferably includes one or more of triethylamine, pyridine, or N,N-diisopropylethylamine, pyridine, potassium carbonate, cesium carbonate, and potassium phosphate, more preferably pyridine. Preferably, the temperature of the second ring-closing reaction is 80–120°C, more preferably 100°C, and the time is preferably 6–12 hours, more preferably 8 hours. After the second ring-closing reaction, the resulting reaction solution is preferably poured into water, and after the yellow solid precipitates, it is filtered to obtain compound f.

[0088] In this invention, compound f undergoes a first hydrolysis reaction to obtain compound g. In this invention, the first hydrolysis reaction is preferably carried out under an alkaline environment. In this invention, the base used in the first hydrolysis reaction preferably includes one or more of triethylamine, pyridine, or N,N-diisopropylethylamine, potassium carbonate, sodium hydroxide, potassium hydroxide, cesium carbonate, and potassium phosphate, more preferably sodium hydroxide. In this invention, the molar ratio of compound f to the base is preferably 1:(4-10), more preferably 1:5.

[0089] In this invention, the first hydrolysis reaction is preferably carried out in the presence of water. The temperature of the first hydrolysis reaction is preferably 50–70°C, more preferably 60°C, and the time is preferably 4–6 hours, more preferably 5 hours. After the first hydrolysis reaction, the pH is adjusted to 1 with 6N HCl, and the mixture is filtered to obtain compound g.

[0090] In this invention, compound g undergoes a first oxidation reaction to obtain compound h. In this invention, the oxidant used in the first oxidation reaction is cerium ammonium nitrate; in this invention, the molar ratio of compound g to the oxidant is preferably 1:(2-5), more preferably 1:3.

[0091] In this invention, the organic solvent used in the first oxidation reaction is preferably acetonitrile. In this invention, the temperature of the first oxidation reaction is preferably -10 to 0°C, more preferably 0°C, and the time is preferably 4 to 6 hours, more preferably 5 hours. After the first oxidation reaction, the solvent is removed under reduced pressure. In this invention, the resulting reaction solution is preferably poured into water, and after the solid precipitates, it is filtered to obtain compound h. In this invention, when R1 is a hydroxyl group, compound h is a BBI608 derivative having the structure shown in Formula I.

[0092] When R1 is one of the following: a substituted or unsubstituted C1-C10 alkylamine, a substituted or unsubstituted C1-C10 alkanolamine, a substituted or unsubstituted C3-C7 cycloalkylamine, a substituted or unsubstituted 5-10 membered heterocyclic amine, or a heteroaromatic amine, in the presence of 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate and N,N-diisopropylethylamine, compound h undergoes a first acylation reaction with compound 3 to obtain a BBI608 derivative having the structure shown in Formula I. In this invention, compound 3 is a substituted or unsubstituted C1-C10 alkylamine, a substituted or unsubstituted C1-C10 alkanolamine, a substituted or unsubstituted C3-C7 cycloalkylamine, a substituted or unsubstituted 5-10 membered heterocyclic amine, or a heteroaromatic amine, with the general structural formula R1H. In this invention, the molar ratio of compound h to compound 3 is preferably 1:(1.5 to 2.0), and more preferably 1:2.

[0093] In this invention, the molar ratio of compound h, 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate, and N,N-diisopropylethylamine is preferably 1:(1.2-1.7):(2-3), more preferably 1:1.2:2.4. In this invention, the temperature of the first acylation reaction is preferably 25-35°C, more preferably 30°C; the time is preferably 1-3 h, more preferably 2 h. After the first acylation reaction, the resulting reaction solution is preferably poured into water, extracted with dichloromethane, and then successively washed with 1 mol / L hydrochloric acid, 5 wt% sodium carbonate aqueous solution, water, saturated brine, and dried with anhydrous sodium sulfate. The solution is then filtered, and the filtrate is concentrated under reduced pressure and subjected to column chromatography to obtain BBI608 derivatives with the structure shown in Formula I.

[0094] In this invention, the BBI608 derivative having the structure shown in Formula I is preferably compound SC-1 to 11, SC-13 to 17, SC-22 to 25 or SC-27.

[0095] In this invention, the reaction route of the BBI608 derivative having the structure shown in Formula I is as follows: Figure 1 As shown.

[0096] (ii) When the BBI608 derivative has the structure shown in Formula II, the preparation method includes the following steps:

[0097] Compound g and compound 4 undergo a second acylation reaction to yield compound i;

[0098] Compound i undergoes a first electrophilic substitution reaction with a Grignard reagent to obtain compound j;

[0099] Compound j undergoes a second reduction reaction to yield compound k;

[0100] The compound k undergoes a second oxidation reaction to yield compound l; when R1 is a hydroxyl group, compound l is a BBI608 derivative having the structure shown in Formula II;

[0101] When R1 is one of the following: a substituted or unsubstituted C1-C10 alkylamine group, a substituted or unsubstituted C1-C10 alkylolamine group, a substituted or unsubstituted C3-C7 cycloalkylamine group, a substituted or unsubstituted 5-10 membered heterocyclic amine group, or a heteroaromatic amine group, the compound l undergoes a third nucleophilic substitution reaction with a chlorinating reagent to give compound m.

[0102] The compound m undergoes a fourth nucleophilic substitution reaction with the compound 3 to obtain a BBI608 derivative having the structure shown in Formula II.

[0103] Compound 4; R1H compound 3;

[0104]

[0105]

[0106] In this invention, compound g and compound 4 undergo a second acylation reaction to obtain compound i. In this invention, the reaction conditions and post-treatment methods used in the second acylation reaction are preferably consistent with those of the first acylation reaction in step (i), and will not be repeated here.

[0107] In this invention, compound i undergoes a first electrophilic substitution reaction with a Grignard reagent to obtain compound j. In this invention, the Grignard reagent is preferably magnesium methyl bromide; the molar ratio of compound i to the Grignard reagent is preferably 1:(1.2-2), more preferably 1:1.5. In this invention, the organic solvent used in the first electrophilic substitution reaction preferably includes one or more of acetonitrile, dichloromethane, and tetrahydrofuran, more preferably tetrahydrofuran. In this invention, the temperature of the first electrophilic substitution reaction is preferably -25 to 0°C, more preferably -10°C, and the time is preferably 2 to 4 hours, more preferably 3 hours. After the first electrophilic substitution reaction, water is added, the solvent is removed under reduced pressure, ethyl acetate is added for extraction, the mixture is dried, filtered, and concentrated to obtain compound j.

[0108] In this invention, compound j undergoes a second reduction reaction to obtain compound k. In this invention, the reducing agent is preferably one or more of sodium borohydride, sodium cyanoborohydride, and sodium borohydride acetate, more preferably sodium borohydride. In this invention, the molar ratio of compound j to the reducing agent is preferably 1:(1.2-2), more preferably 1:1.5. In this invention, the organic solvent used in the second reduction reaction is preferably one or more of acetonitrile, dichloromethane, and tetrahydrofuran, more preferably tetrahydrofuran. In this invention, the temperature of the second reduction reaction is preferably 0-25°C, more preferably 10°C, and the time is preferably 2-4 hours, more preferably 3 hours. After the second reduction reaction, water is added, the solvent is removed under reduced pressure, ethyl acetate is added for extraction, the mixture is dried, filtered, and concentrated to obtain compound k.

[0109] In this invention, compound k undergoes a second oxidation reaction to obtain compound l. In this invention, the reaction conditions and post-treatment methods used in the second oxidation reaction are preferably consistent with those of the first oxidation reaction in step (i), and will not be repeated here. In this invention, when R1 is a hydroxyl group, compound l is a BBI608 derivative having the structure shown in Formula II.

[0110] When R1 is one of the following: a substituted or unsubstituted C1-C10 alkylamine group, a substituted or unsubstituted C1-C10 alkylolamine group, a substituted or unsubstituted C3-C7 cycloalkylamine group, a substituted or unsubstituted 5-10 membered heterocyclic amine group, or a heteroaromatic amine group, compound l undergoes a third nucleophilic substitution reaction with a chlorinating reagent to obtain compound m. In this invention, the chlorinating reagent is preferably thionyl chloride and / or phosphorus trichloride. In this invention, the molar ratio of compound l to the chlorinating reagent is preferably 1:(1.2-2), more preferably 1:1.5. In this invention, the organic solvent used for the third nucleophilic substitution reaction is preferably one or more of acetonitrile, dichloromethane, and tetrahydrofuran, more preferably dichloromethane. In this invention, the temperature of the third nucleophilic substitution reaction is preferably 0-25°C, more preferably 10°C, and the time is preferably 2-4 h, more preferably 3 h. After the third nucleophilic substitution reaction, water is added, the solvent is removed under reduced pressure, ethyl acetate is added for extraction, the mixture is dried, filtered, and concentrated to obtain compound m.

[0111] In this invention, compound m undergoes a fourth nucleophilic substitution reaction with compound 3 to obtain a BBI608-type derivative having the structure shown in Formula II. In this invention, compound 3 is a substituted or unsubstituted C1-C10 alkylamine, a substituted or unsubstituted C1-C10 alkanolamine, a substituted or unsubstituted C3-C7 cycloalkylamine, a substituted or unsubstituted 5-10 membered heterocyclic amine, or a heteroaromatic amine, with the general structural formula R1H.

[0112] In this invention, the fourth substitution reaction is preferably carried out under alkaline conditions. The base used in the fourth nucleophilic substitution reaction preferably includes one or more of triethylamine, pyridine, or N,N-diisopropylethylamine, pyridine, potassium carbonate, cesium carbonate, and potassium phosphate, more preferably potassium carbonate. In this invention, the molar ratio of compound m, compound 3, and base is preferably 1.0:(1.2-1.5):(:2-5), more preferably 1:1.3:3. In this invention, the temperature of the fourth nucleophilic substitution reaction is preferably 50-80°C, more preferably 60°C, and the time is preferably 2-4 hours, more preferably 3 hours. After the fourth nucleophilic substitution reaction, in this invention, the resulting reaction solution is preferably poured into water, the solvent is removed under reduced pressure, ethyl acetate is added for extraction, dried, then filtered, concentrated, and column chromatography is performed to obtain BBI608 derivatives having the structure shown in II.

[0113] In this invention, the BBI608 derivative having the structure shown in II is preferably SD-1 to 6.

[0114] In this invention, the reaction route of the BBI608 derivative having the structure shown in Formula II is as follows: Figure 2 As shown.

[0115] (iii) When the BBI608 derivative has the structure shown in Formula III, the preparation method includes the following steps:

[0116] Compound e and compound 5 undergo a third ring-cladding reaction to yield compound n;

[0117] The compound n undergoes a second hydrolysis reaction to yield compound o;

[0118] Compound o undergoes a third oxidation reaction to yield compound p;

[0119] When R1 is a hydroxyl group, compound p is a BBI608-type derivative having the structure shown in Formula III;

[0120] When R1 is one of the following: substituted or unsubstituted C1-C10 alkylamine, substituted or unsubstituted C1-C10 alkylamino, substituted or unsubstituted C3-C7 cycloalkylamine, substituted or unsubstituted 5-10 membered heterocyclic amine or heteroaromatic amine, in the presence of 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate and N,N-diisopropylethylamine, compound p and compound 3 undergo a third acylation reaction to obtain BBI608 derivatives having the structure shown in Formula III;

[0121]

[0122] In this invention, compound e and compound 5 undergo a third ring-closing reaction to obtain compound n. In this invention, the molar ratio of compound e to compound 5 is preferably 1:(2-5), more preferably 1:3. In this invention, the organic solvent preferably includes acetonitrile, acetone, and tetrahydrofuran, and the reaction is preferably carried out in the presence of tetrahydrofuran. In this invention, the temperature of the third ring-closing reaction is preferably 60-90°C, more preferably 80°C, and the time is preferably 2-8 hours, more preferably 3 hours. After the third ring-closing reaction, the resulting reaction solution is preferably concentrated under reduced pressure to obtain compound n.

[0123] In this invention, compound n undergoes a second hydrolysis reaction to obtain compound o. In this invention, the reaction conditions and post-treatment methods used in the second hydrolysis reaction are preferably consistent with those of the first hydrolysis reaction in step (i), and will not be repeated here.

[0124] In this invention, compound o undergoes a third oxidation reaction to yield compound p. When R1 is a hydroxyl group, compound p is a BBI608-type derivative having the structure shown in Formula III.

[0125] In this invention, the reaction conditions and post-treatment methods used in the third oxidation reaction are preferably the same as those in the first oxidation reaction in step (i), and will not be repeated here.

[0126] When R1 is one of the following: a substituted or unsubstituted C1-C10 alkylamine, a substituted or unsubstituted C1-C10 alkanolamine, a substituted or unsubstituted C3-C7 cycloalkylamine, a substituted or unsubstituted 5-10 membered heterocyclic amine, or a heteroaromatic amine, in the presence of 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate and N,N-diisopropylethylamine, compound p and compound 3 undergo a third acylation reaction to obtain BBI608 derivatives having the structure shown in Formula III. In this invention, compound 3 is a substituted or unsubstituted C1-C10 alkylamine, a substituted or unsubstituted C1-C10 alkanolamine, a substituted or unsubstituted C3-C7 cycloalkylamine, a substituted or unsubstituted 5-10 membered heterocyclic amine, or a heteroaromatic amine, with the general structural formula R1H. In this invention, the molar ratio of compound p to compound 3 is preferably 1:(1.5-2.0), more preferably 1:2. In this invention, the molar ratio of compound p, 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate, and N,N-diisopropylethylamine is preferably 1:(1.2-1.7):(2-3), more preferably 1:1.2:2.4. In this invention, the temperature of the third acylation reaction is preferably 25-35°C, more preferably 30°C; the time is preferably 1-3 h, more preferably 2 h. After the third acylation reaction, the resulting reaction solution is preferably poured into water, extracted with dichloromethane, and then successively washed with 1 mol / L hydrochloric acid, 5 wt% sodium carbonate aqueous solution, water, saturated brine, and dried with anhydrous sodium sulfate. The solution is then filtered, and the filtrate is concentrated under reduced pressure and subjected to column chromatography to obtain BBI608 derivatives with the structure shown in Formula III.

[0127] In this invention, the BBI608 derivatives having the structure shown in III are preferably SG-1 to 8, SG-12, SG-13, SG-15, SG-16, SG-18, SG-19, and SG-22 to 24.

[0128] In this invention, the reaction route of the BBI608 derivative having the structure shown in Formula III is as follows: Figure 3 As shown.

[0129] This invention provides the application of the above-mentioned BBI608 derivatives in the preparation of drugs for treating STAT3-mediated diseases.

[0130] In this invention, the STAT3-mediated diseases include one or more of colorectal cancer, lung cancer, melanoma, breast cancer, prostate cancer, kidney cancer, ovarian cancer, liver cancer, pancreatic cancer, multiple myeloma, and leukemia.

[0131] This invention provides a medicament for treating STAT3-mediated diseases, the active ingredient comprising the aforementioned BBI608 derivatives and their pharmaceutically acceptable salts. In this invention, the salt is an organic or inorganic acid salt of the aforementioned BBI608 derivatives, wherein the inorganic acid is preferably one or more of hydrochloric acid, hydrobromic acid, and sulfuric acid, and the organic acid is preferably one or more of maleic acid, citric acid, fumaric acid, and tartaric acid.

[0132] The following detailed description, in conjunction with embodiments, illustrates the BBI608 derivatives provided by this invention, their preparation methods, and their application in the preparation of drugs for treating STAT3-mediated diseases. However, these descriptions should not be construed as limiting the scope of protection of this invention.

[0133] Example 1 Synthesis of 3,6-dimethoxy-2-nitrobenzaldehyde

[0134] Add concentrated nitric acid (8 mL, 179.02 mmol) to a 100 mL three-necked flask, place the reaction solution in a cold trap, and cool to 0 °C. ℃ Ac₂O (8 mL, 84.24 mmol) was slowly added, followed by 2,5-dimethoxybenzaldehyde (4 g, 24.07 mmol). The addition was exothermic, and the reaction temperature was controlled at approximately 0°C. The addition was completed over 20 minutes. After reacting at room temperature for 1 hour, TLC analysis confirmed the reaction was complete. The reaction solution was poured into 50 mL of ice water and washed once with DCM (40 mL × 3), saturated Na₂CO₃ (20 mL × 4), and saturated saline (10 mL). The solution was dried over anhydrous magnesium sulfate for 4 hours, filtered, and concentrated to obtain 4.78 g of crude product. The sample was mixed with 5.2 g of silica gel, packed into a 20 g silica gel column, and the mobile phase was EA:PE = 1:10 to obtain the above impurity spot 2.30 g (2.30 g, 45.28%). The mobile phase was EA:PE = 1:10 to obtain (2.52 g, 49.62%). 1 H NMR (400MHz, CDCl3): δ (ppm) 10.25 (s, 1H), 7.69 (d, J = 9.4Hz, 1H), 7.48 (d, J = 9.4Hz, 1H), 3.95 (s, 3H), 3.86 (s, 3H).

[0135] Example 2 Synthesis of methyl 4,7-dimethoxybenzo[b]thiophene-2-carboxylate

[0136] 3,6-Dimethoxy-2-nitrobenzaldehyde (1.0 g, 4.73 mmol), methyl mercaptoacetate (0.55 g, 5.21 mmol), and K₂CO₃ (0.72 g, 5.21 mmol) were added to a 100 mL single-necked flask. The mixture was heated to 70 °C and reacted for 8 h. TLC analysis showed that the reaction was complete. The reaction solution was poured into 50 mL of ice water, stirred for 0.5 h, filtered, and concentrated to obtain (0.80 g, 67.22%). 1 H NMR (400MHz, CDCl3): δ (ppm) 8.20 (s, 1H), 6.76 (d, J = 8.4Hz, 1H), 6.66 (d, J = 8.4Hz, 1H), 3.95 (s, 3H), 3.93 (s, 3H), 3.91 (s, 3H). ESI-MS: m / z 253.00[M+H] + .

[0137] Example 3 Synthesis of methyl 4,7-dimethoxy-5,6-dinitrobenzo[b]thiophene-2-carboxylate

[0138] 10 mL of concentrated nitric acid was added to a 50 mL three-necked flask. The reaction solution was placed in a cold trap and cooled to 0 °C. Methyl 4,7-dimethoxybenzo[b]thiophene-2-carboxylic acid (1.0 g, 3.96 mmol) was slowly added. The reaction was exothermic during the addition. The reaction temperature was controlled at around 0 °C. The addition was completed in 15 minutes. After reacting at room temperature for 1 hour, the reaction was confirmed to be complete by TLC. The reaction solution was poured into 50 mL of ice water and filtered to obtain a yellow solid (0.65 g, 48.0%). 1 H NMR (400MHz, CDCl3): δ (ppm) 8.26 (s, 1H), 4.17 (s, 3H), 4.16 (s, 3H), 4.02 (s, 3H). ESI-MS: m / z 3433.00 [M+H] + 365.0 [M+Na] + .

[0139] Example 4 Synthesis of methyl 5,6-diamino-4,7-dimethoxybenzo[b]thiophene-2-carboxylate

[0140] 4,7-Dimethoxy-5,6-dinitrobenzo[b]thiophene-2-carboxylic acid methyl ester (0.9 g, 2.63 mmol), EA (20 mL), and Pt (0.18 g) were added to a 100 mL single-necked flask. H2 was introduced, and the mixture was stirred at 50 °C for 16 h. The reaction was confirmed to be complete by TLC. The reaction solution was filtered through diatomaceous earth and concentrated under reduced pressure to obtain the product (0.60 g, 81.08%). 1H NMR (400MHz, DMSO-d6): δ7.84(s,1H),7.52(s,1H),4.67(s,2H),3.83(s,3H),3.77(s,3H),3.75(s,3H).ESI-MS:m / z282.07[M+H] + .

[0141] Example 5 Synthesis of methyl 4,8-dimethoxythiopheno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-6-carboxylate

[0142] Methyl 5,6-diamino-4,7-dimethoxybenzo[b]thiophene-2-carboxylate (0.73 g, 2.58 mmol), Py (10 mL), and PhNSO (0.79 g, 5.19 mmol) were added to a 100 mL single-necked flask. The mixture was stirred at 120 °C for 12 h. The reaction was confirmed to be complete by TLC. The reaction solution was poured into 20 mL of 3NHCl, and a red solid precipitated out. The product (0.72 g, 92.30%) was obtained by filtration. 1 H NMR (400MHz, DMSO-d6): δ8.24(s,1H),4.50(s,3H),4.23(s,3H),3.97(s,3H).ESI-MS:m / z311.0[M+H] + 333.0[M+Na] + .

[0143] Example 6 Synthesis of 4,8-dimethoxythiopheno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-6-carboxylic acid

[0144] 4,8-Dimethoxythiophene[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-6-carboxylate (3.37 g, 10.87 mmol), THF (40 mL), LiOH (0.91 g, 21.74 mmol), and water (40 mL) were added to a 250 mL single-necked flask. The mixture was stirred at 25 °C for 2 h. The reaction was confirmed to be complete by TLC. The reaction solution was poured into 80 mL of 3N HCl, and a red solid precipitated. The product (1.60 g, 64.78%) was obtained by filtration. 1 H NMR(400MHz, CDCl3): δ8.36(s,1H),4.54(s,3H),4.45(s,3H).ESI-MS:m / z 264.1[MH] - .

[0145] Example 7 Synthesis of 4,8-dioxo-4,8-dihydrothiopheno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-6-carboxylic acid

[0146] Add 0.2 g (0.675 mmol) of 4,8-dimethoxythiophene[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazol-6-carboxylic acid, acetonitrile (5 mL), and water (5 mL) to a 50 mL three-necked flask. Cool to 0 °C and add dropwise a solution of 5 mL of acetonitrile containing CAN (0.85 g, 1.82 mmol), completing the addition over 10 min. TLC analysis confirmed the reaction was complete. Add 10 mL of water to the reaction mixture, filter, and obtain the product (0.16 g, 90.0%). ESI-MS: m / z 264.1 [MH] - .

[0147] Example 8 Synthesis of 6-(4-methylpiperazine-1-carbonyl)thieno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-4,8-dione (SC-1)

[0148] Add 0.19 g (0.71 mmol) of 8-dioxo-4,8-dihydrothiopheno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazol-6-carboxylic acid, 5 mL of dry DCM, 0.32 g (0.85 mmol) of HATU, and 0.22 g (1.7 mmol) of DIPEA to a 25 mL single-necked flask. After 0.5 h, the reaction was completed by TLC. Remove the solvent under reduced pressure, and add 71 mg of N-methylpiperazine dropwise. 1.07 mmol), after 1 h, the reaction was detected by TLC to be complete. The solvent was removed under reduced pressure, and 10 mL of water was added to the reaction solution. The reaction solution was poured into 10 mL of water, washed once with DCM (10 mL × 3) and saturated saline (20 mL), dried over anhydrous magnesium sulfate for 4 h, filtered, concentrated, and 0.22 g of pale yellow solid SC-1 was obtained. The sample was mixed with 0.30 g of silica gel, packed into a column with 1 g of silica gel, and the mobile phase was EA:PE = 2:1 to obtain SC-1 (60 mg, 25%). mp 225-227℃ 1 H NMR (400MHz, CDCl3): δ (ppm) 7.85 (s, 1H), 3.78 (s, 4H), 2.51 (t, J = 8.4Hz, 4H), 2.36 (s, 3H). 13 C NMR (100MHz, CDCl3): δ171.64,170.41,160.65,156.60,156.47,147.78,147.39,142.52,126.53,65.51,54.80,45.93.

[0149] Example 9: Synthesis of N,N-dimethyl-4,8-dioxo-4,8-dihydrothiophene[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-6-carboxamide (SC-2)

[0150] Following the method described in Example 8, a white solid, SC-2, was obtained from dimethylamine as a raw material. The yield was 6 mg, with a recovery rate of 30.0%, and the mp value was 185-187°C. 1 H NMR (400MHz, CDCl3): δ (ppm) 7.92 (s, 1H), 3.30 (s, 3H), 3.19 (s, 3H) 13 C NMR (100MHz, CDCl3): δ171.74,170.51,161.74,156.67,156.54,148.69,147.50,142.61,126.78,39.65,36.66.

[0151] Example 10 Synthesis of 6-(pyrrolidine-1-carbonyl)thieno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-4,8-dione (SC-3)

[0152] Following the method described in Example 8, a white solid, SC-3, was obtained from tetrahydropyrrole as a raw material. The yield was 72 mg, the recovery rate was 32.0%, and the mp value was 205-207°C. 1 H NMR (400MHz, CDCl3): δ (ppm) 8.04 (s, 1H), 3.84 (t, J = 6.44Hz, 2H), 3.72 (t, J = 6.72Hz, 2H), 2.12-2.00 (m, 4H) 13 C NMR (100MHz, CDCl3): δ171.87,170.66,159.32,156.71,156.58,150.39,147.76,142.95,126.63,49.22,47.92,26.76,24.00.

[0153] Example 11 Synthesis of (R)-1-(4,8-dioxo-4,8-dihydrothiopheno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-6-carbonyl)piperidine-3-carboxylic acid ethyl ester (SC-4)

[0154] Following the method described in Example 8, a white solid, SC-4, was obtained from (R)-piperidine-3-carboxylic acid ethyl ester as a raw material. The yield was 89 mg, the recovery rate was 31%, and the mp was 195-197 °C. 1H NMR (400MHz, CDCl3): δ (ppm) 7.88 (s, 1H), 4.49-4.15 (m, 3H), 3.99-3.96 (m, 1H), 3.67-3.40 (m, 2H), 2.64-2.60(m,1H),2.16-2.13(m,1H),1.87-1.86(m,2H),1.69-1.64(m,1H),1.27(t,J=7.1Hz,3H). 13 C NMR (100MHz, CDCl3): δ172.42,171.64,170.44,161.03,156.63,156.50,147.99,147.33,142.54,126.43,61.04,41.22,27.10,14.19.

[0155] Example 12 Synthesis of 6-(4-acetylpiperazine-1-carbonyl)thieno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-4,8-dione (SC-5)

[0156] Following the method described in Example 8, a white solid, SC-5, was obtained from 1-(piperazin-1-yl)ethane-1-one as a raw material. The yield was 77 mg, with a recovery rate of 29%, and the mp value was 200-202 °C. 1 H NMR (400MHz, CDCl3): δ (ppm) 8.04 (s, 1H), 3.84 (t, J = 6.44Hz, 2H), 3.72 (t, J = 6.72Hz, 2H), 2.12-2.00 (m, 4H) 13 C NMR (100MHz, CDCl3): δ171.87,170.66,159.32,156.71,156.58,150.39,147.76,142.95,126.63,49.22,47.92,26.76,24.00.

[0157] Example 13 Synthesis of (S)-1-(4,8-dioxo-4,8-dihydrothieno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-6-carbonyl)piperidine-3-carboxylic acid ethyl ester (SC-6)

[0158] Following the method described in Example 8, a white solid, SC-6, was obtained from (S)-piperidine-3-carboxylic acid ethyl ester as a raw material. The yield was 80 mg, with a yield of 28% and an mp of 196-197°C. 1H NMR (400MHz, CDCl3): δ (ppm) 7.88 (s, 1H), 4.49-4.15 (m, 3H), 3.99-3.96 (m, 1H), 3.67-3.40 (m, 2H), 2.64-2.60(m,1H),2.16-2.13(m,1H),1.87-1.86(m,2H),1.69-1.64(m,1H),1.27(t,J=7.1Hz,3H). 13 C NMR (100MHz, CDCl3): δ172.42,171.64,170.44,161.03,156.63,156.50,147.99,147.33,142.54,126.43,61.04,41.22,27.10,14.19.

[0159] Example 14 Synthesis of 4,8-dioxo-N-(2-(piperidin-1-yl)ethyl)-4,8-dihydrothiopheno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-6-carboxamide (SC-7)

[0160] Following the method described in Example 8, a white solid, SC-7, was obtained from 2-(piperidin-1-yl)ethane-1-amine as a raw material. The yield was 72 mg, the recovery rate was 27%, and the mp value was 186-187 °C. 1 H NMR (400MHz, CDCl3): δ (ppm) 7.88 (s, 1H), 7.59 (s, 1H), 3.59 (d, J = 3.9Hz, 2H ),2.67(t,J=5.6Hz,2H),2.55(s,4H),1.70-1.66(m,4H),1.54-1.53(m,2H). 13 C NMR (100MHz, CDCl3): δ171.73,170.63,159.68,156.65,156.51,149.81,148.18,143.26,125.44,56.66,54.29,36.42,25.52,23.97.

[0161] Example 15: Synthesis of 5N-(2-hydroxypropyl)-4,8-dioxo-4,8-dihydrothiopheno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-6-carboxamide (SC-8)

[0162] Following the method described in Example 8, a white solid, SC-8, was obtained from 1-aminoprop-2-ol as a raw material. The yield was 57 mg, with a recovery rate of 25%, and the mp value was 198-200 °C. 1H NMR (400MHz, CDCl3): δ (ppm) 9.05 (t, J = 5.6Hz, 1H), 8.47 (s, 1H), 4.80 (d, J = 3.7Hz, 2H), 3.83-3.78 (m, 1H), 3.24-3.20 (m, 2H), 1.09 (d, J = 6.2Hz, 3H). 13 C NMR (100MHz, CDCl3): δ172.83,171.84,160.01,157.60,157.37,149.32,148.28,144.02,126.36,65.31,47.82,21.62.

[0163] Example 16 Synthesis of methyl(4,8-dioxo-4,8-dihydrothieno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-6-carbonyl)-D-valine (SC-9)

[0164] Following the method described in Example 8, a white solid, SC-9, was obtained from D-valine methyl ester as a raw material. The yield was 70 mg, the recovery rate was 26%, and the mp value was 189-191 °C. 1 H NMR (400MHz, CDCl3): δ (ppm) 8.13 (s, 1H), 6.84 (d, J = 8.44Hz, 1H), 4.75 (dd, J = 8.44Hz, 4.8Hz, 1H), 3.82 (s, 3H), 2.34-2.29 (m, 1H), 1.04-1.01 (m, 6H). 13 C NMR (100MHz, CDCl3): δ171.92,171.63,170.49,159.54,156.59,156.48,148.69,148.66,143.13,125.51,57.95,52.62,31.63,18.97,17.93.

[0165] Example 17 Synthesis of 6-(piperidine-1-carbonyl)thieno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-4,8-dione (SC-10)

[0166] Following the method described in Example 8, a white solid, SC-10, was obtained from piperidine as a raw material. The yield was 71 mg, the recovery rate was 30%, and the mp value was 193-195 °C. 1 H NMR (400MHz, CDCl3): δ (ppm) 7.83 (s, 1H), 3.68 (s, 4H), 1.76-1.69 (m, 6H) 13C NMR (100MHz, CDCl3): δ171.69, 170.45, 160.57, 156.67, 156.56, 148.50, 147.16, 142.60, 126.19, 24.34.

[0167] Example 18 Synthesis of methyl(4,8-dioxo-4,8-dihydrothieno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-6-carbonyl)-D-proline (SC-11)

[0168] Following the method described in Example 8, a white solid, SC-11, was obtained from D-proline methyl ester as a raw material. The yield was 64 mg, the recovery rate was 24%, and the mp was 125-127 °C. 1 H NMR (400MHz, CDCl3): δ (ppm) 8.11 (s, 1H), 4.72 (m, 1H), 4.06 (m, 1H), 3.93 (m, 1H), 2.35 (m, 1H), 2.24 (m, 1H), 2.14-2.11 (m, 2H), 13 C NMR (100MHz, CDCl3): δ171.76x2,170.59,159.67,156.69,156.54,149.10,148.17,142.91,127.33,60.66,52.56,49.58,28.85,22.51.

[0169] Example 19 Synthesis of N,N-diethyl-4,8-dioxo-4,8-dihydrothiopheno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-6-carboxamide (SC-13)

[0170] Following the method described in Example 8, a white solid, SC-13, was obtained from diethylamine as a raw material. The yield was 68 mg, the recovery rate was 30%, and the mp value was 125-127 °C. 1 H NMR (400MHz, CDCl3): δ (ppm) 7.86 (s, 1H), 3.57 (q, J = 7.12Hz, 4H), 1.31 (t, J = 6.8Hz, 6H) 13 C NMR (100MHz, CDCl3): δ171.67,170.48,161.16,156.67,156.55,149.05,147.09,142.74,125.76,14.45,12.95.

[0171] Example 20 Synthesis of 6-(4-(bis(4-fluorophenyl)methyl)piperazine-1-carbonyl)thieno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-4,8-dione (SC-14)

[0172] Following the method described in Example 8, a white solid, SC-14, was obtained from 1-(bis(4-fluorophenyl)methyl)piperazine as a raw material. The yield was 76 mg, the recovery rate was 21%, and the mp value was 245-247 °C. 1 H NMR (400MHz, CDCl3): δ (ppm) 7.80 (s, 1H), 7.36 (s, 4H), 7.00 (t, J = 8.52Hz, 4H), 4.31 (s, 1H), 3.77 (s, 4H), 2.48 (s, 4H) 13 C NMR (100MHz, CDCl3): δ171.57,170.73,163.29,160.85,160.57,156.60,156.48,147.73,147.41,142.50,142 .50,137.22,192.30,192.30,129.22,129.22,126.54,115.85,115.85,115.64,115.64,74.19,51.65,51.65.

[0173] Example 21 Synthesis of 4-(4,8-dioxo-4,8-dihydrothiopheno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-6-carboxamido)piperidine-1-carboxylic acid tert-butyl ester (SC-15)

[0174] Following the method described in Example 8, a white solid, SC-15, was obtained from tert-butyl 4-aminopiperidine-1-carboxylate as a raw material. The yield was 73 mg, the recovery rate was 23%, and the mp was 193-195 °C. 1 H NMR (400MHz, CDCl3): δ (ppm) 8.08 (s, 1H), 6.35 (d, J = 6.68, 1H), 4.15 (d, J = 11. 76,2H),2.94-2.88(m,2H),2.06(s,1H),2.03(s,1H),1.90(s,3H),1.48(s,9H) 13C NMR (100MHz, CDCl3): δ171.77,170.51,158.95,156.56,156.49,154.70,149.50,148 .50,143.11,125.06,79.99,53.43,48.16,42.68,31.92,31.23,28.44,28.25,24.12.

[0175] Example 22 Synthesis of 6-(4-diphenylmethylpiperazine-1-carbonyl)thieno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-4,8-dione (SC-16)

[0176] Following the method described in Example 8, a white solid, SC-16, was obtained from 1-benzylpiperazine as a raw material. The yield was 74 mg, with a recovery rate of 21%, and the mp value was 232-235 °C. 1 H NMR (400MHz, CDCl3): δ (ppm) 7.80 (s, 1H), 7.43 (s, 2H), 7.42 (s, 2H), 7.30 (t, J=7.28,4H),7.23(s,1H),7.21(s,1H),4.31(s,1H),3.77(s,4H),2.51(s,4H) 13 C NMR (100MHz, CDCl3): δ171.57,170.38,160.52,156.60,156.48,147.91,147.34,142.48,141.71,128.74,127.86,127.37,126.49,75.82,51.78.

[0177] Example 23 Synthesis of 4-(4,8-dioxo-4,8-dihydrothiopheno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-6-carbonyl)piperazine-1-carboxylic acid tert-butyl ester (SC-17)

[0178] Following the method described in Example 8, a white solid, SC-17, was obtained from piperazine-1-carboxylic acid tert-butyl ester as a raw material. The yield was 68 mg, the recovery rate was 22%, and the mp was 188-190 °C. 1 H NMR (400MHz, CDCl3): δ (ppm) 7.85 (s, 1H), 3.74 (s, 4H), 3.57-3.54 (m, 4H), 1.49 (s, 9H) 13C NMR (100MHz, CDCl3): δ171.55,170.36,161.00,156.59,156.45,154.36,147.59,147.30,142.56,126.68,80.77,43.56,28.36.

[0179] Example 24 Synthesis of 6-(4-benzylpiperidin-1-carbonyl)thieno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-4,8-dione (SC-22)

[0180] Following the method described in Example 8, a white solid, SC-22, was obtained from 4-benzylpiperidine as a raw material. The yield was 69 mg, with a recovery rate of 23% and an mp of 202-204 °C. 1 H NMR (400MHz, CDCl3): δ7.81(s,1H),7.32-7.30(m,2H),7.23-7.20(m,1H),7.16-7.14(m,2H), 4.59-4.09(m,2H),3.02(s,2H),2.61(d,J=7.0Hz,2H),1.93-1.79(m,3H),1.37-1.24(m,2H). 13 C NMR (100MHz, CDCl3): δ171.68,170.44,160.58,156.63,156.51,148.35,147.18,14 2.54,139.51,129.08,128.55,126.26,126.24,48.58,42.78,38.15,32.63,31.73.

[0181] Example 25 Synthesis of 1-(4,8-dioxo-4,8-dihydrothiopheno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-6-carbonyl)piperidine-4-carboxylic acid ethyl ester (SC-23)

[0182] Following the method described in Example 8, a white solid, SC-23, was obtained from piperidine-4-carboxylic acid ethyl ester as a raw material. The yield was 63 mg, the recovery rate was 23%, and the mp value was 186-188 °C. 1 H NMR (400MHz, CDCl3): δ7.84(s,1H),4.36-4.16(m,4H),3.17(s,2H),2.68-2.23m,1H),2.06-2.04(m,2H),1.88-1.78(m,2H),1.27(t,J=7.1Hz,3H). 13C NMR (100MHz, CDCl3): δ173.62,171.03,170.41,160.74,156.62,156.49,147.88,147.35,142.56,126.38,60.92,40.61,28.03,14.22.

[0183] Example 26 Synthesis of (S)-(1-(4,8-dioxo-4,8-dihydrothieno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-6-carbonyl)piperidin-3-yl)tert-butyl carbamate (SC-24)

[0184] Following the method described in Example 8, a white solid, SC-24, was obtained from (S)-piperidin-3-ylcarbamate tert-butyl ester as a raw material. The yield was 73 mg, the recovery rate was 23%, and the mp was 191-193 °C. 1 H NMR (400MHz, CDCl3): δ7.96(s1H),4.62(s,1H),4.00(s,1H),3.75(s,2H),3.61(s,1H) ),3.50-3.49(m,1H),2.04-2.00(m,1H),1.86-1.83(m,1H),1.66(s,2H),1.92(s,9H). 13 C NMR (100MHz, CDCl3): δ171.54,170.46,161.43,156.68,156.47,155.02,147.88, 147.39,142.65,126.90,80.17,52.46,48.18,46.97,43.63,29.90,28.34,22.40.

[0185] Example 27 Synthesis of 1-(4,8-dioxo-4,8-dihydrothiopheno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-6-carbonyl)piperidine-4-carboxylic acid methyl ester (SC-25)

[0186] Following the method described in Example 8, a white solid, SC-25, was obtained from piperidine-4-carboxylic acid methyl ester as a raw material. The yield was 64 mg, the recovery rate was 23%, and the mp value was 184-186 °C. 1 H NMR (400MHz, CDCl3): δ7.83(s,1H),4.30(s,2H),3.73(s,3H),3.28(s,2H),3.27(s,2H),2.72-2.65(m,1H),2.08-2.04(m,2H),1.88-1.79(m,2H). 13C NMR (100MHz, CDCl3): δ174.06,171.61,170.41,160.71,156.61,156.48,147.82,147.34,142.55,126.37,52.08,47.06,42.60,40.47,28.17.

[0187] Example 28 Synthesis of 4-(4,8-dioxo-4,8-dihydrothiopheno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-6-carbonyl)-1-(2,2-diphenylacetyl)piperazine-2-carbonyl)-L-valine methyl ester (SC-27)

[0188] Following the method described in Example 8, a white solid, SC-27, was obtained from (1-(2,2-diphenylacetyl)piperazine-2-carbonyl)-L-valine methyl ester, with a yield of 97 mg and a recovery rate of 20%, at mp 255-257 °C. 1 H NMR(400MHz, CDCl3): δ8.00(s 1H),7.38-7.22m,10H),6.66(br,1H),5.33-5.26(m,2H),4.69-4.66(m,1H),4.48-4.43(m,1H),4.32-4.30(m,1H),3.97-3.94(m,1H ),3.80-3.71(m,3H),3.49-3.40(m,2H),2.90-2.77(m,1H),2.26-2.11(m,1H),1.62(s,1H),1.33-1.21,(m,1H),0.90-0.77(m,6H). 13 C NMR (100MHz, CDCl3): δ173.37,172.08,171.66,171.62,171.16,170.51, 156.66,156.50,147.75,147.69,142.40,138.35,138.30,138.16,129.06 ,128.92,128.89,128.86,128.83,128.71,127.76,127.68,127.50,127.0 5,60.40,57.30,55.22,55.17,52.31,43.04,42.97,30.39,19.14,19.07.

[0189] Example 30: Synthesis of (4,8-dimethoxythiopheno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazol-6-yl)(morpholino) methyl ketone

[0190] Add 0.2 g (0.67 mmol) of 8-dimethoxythieno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-6-carboxylic acid to a 25 mL single-necked flask, along with 10 mL of dry DCM and 0.12 g (1.01 mmol). After 2 h, the reaction was completed by TLC. The thionyl chloride was removed under reduced pressure, and then 10 mL of dry DCM and 0.10 g (1.0 mmol) of TEA were added. 1 mmol), after 1 h TLC detection of complete reaction, 10 mL water was added to the reaction solution, the reaction solution was poured into 10 mL water, washed once with DCM (10 mL × 3) and saturated saline (20 mL), dried with anhydrous magnesium sulfate for 4 h, filtered, concentrated, and a pale yellow solid M-11 0.30 g was obtained, mixed with 0.40 g silica gel, packed with 1 g silica gel, and the mobile phase was EA:PE = 1:1 to obtain (0.20, 83.0%). 1 H NMR (400MHz, CDCl3): δ (ppm) 7.63 (s, 1H), 4.45 (s, 3H), 4.43 (s, 3H), 3.80 (d, J = 2.8Hz, 4H), 3.78 (s, 4H).

[0191] Example 31 Synthesis of 1-(4,8-dimethoxythieno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazol-6-yl)ethane-1-one

[0192] Add (4,8-dimethoxythiopheno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazol-6-yl)(morpholino) methyl ketone (0.1 g, 0.27 mmol) to a 50 mL three-necked flask, redistill THF (10 mL), cool to -30 °C, and slowly add methyl magnesium bromide (39 mg, 0.328 mmol). After the addition is complete, move to room temperature and react. TLC detection after 2 h is as follows: Add 10 mL of water to the reaction solution, pour the reaction solution into 10 mL of water, wash once with DCM (10 mL × 3) and saturated saline (20 mL), dry with anhydrous magnesium sulfate for 4 h, filter, concentrate, and obtain a pale yellow solid M-12 0.30 g. Mix with 0.40 g silica gel, pack 1 g silica gel into a column, and use EA:PE = 1:30 as the mobile phase to obtain (50 mg, 63.29%). 1 H NMR (400MHz, CDCl3): δ (ppm) 8.11 (s, 1H), 4.55 (s, 3H), 4.41 (s, 3H), 2.69 (s, 3H).

[0193] Example 31 Synthesis of 1-(4,8-dimethoxythiopheno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazol-6-yl)ethane-1-ol

[0194] Add 1-(4,8-dimethoxythieno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazol-6-yl)ethane-1-one (0.2 g, 0.68 mmol) to a 50 mL three-necked flask, redistill THF (10 mL), cool to 0 °C, slowly add sodium borohydride (31 mg, 0.82 mmol), and after the addition is complete, move to room temperature for reaction. After 1 h, the TLC detection is as follows: Add 10 mL of saturated ammonium chloride water to the reaction solution, wash once with DCM (15 mL × 3) and saturated saline (20 mL), dry with anhydrous magnesium sulfate for 4 h, filter, concentrate, and obtain a pale yellow solid (0.15 g, 75.0%). 1 H NMR (400MHz, CDCl3): δ (ppm) 7.32 (d, J = 0.9Hz, 1H), 5.18-5.13 (m, 1H), 4.41 (s, 3H), 4.37 (s, 3H), 1.68 (d, J = 6.5Hz, 3H), ESI-MS: m / z 297 [M+H] + 319.0 [M+Na] + .

[0195] Example 32 Synthesis of 6-(1-hydroxyethyl)thiopheno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-4,8-dione

[0196] Add 1-(4,8-dimethoxythiopheno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazol-6-yl)ethane-1-ol (1.73 g, 5.84 mmol) and acetonitrile (20 mL) to a 100 mL three-necked flask. Cool to 0 °C and slowly add 20 mL of acetonitrile solution containing CAN (6.29, 13.44 mmol). After the addition is complete, allow the mixture to react at room temperature. After 1 h, the reaction mixture is analyzed by TLC as follows. Add 50 mL of water to the reaction mixture and filter to obtain a pale yellow solid (1.32 g, 85.0%). 1 H NMR (400MHz, CDCl3): δ (ppm) 7.62 (d, J = 0.8Hz, 1H), 5.28-5.23 (m, 1H), 1.69 (d, J = 6.5Hz, 3H), ESI-MS: m / z 267 [M+H] + 289.0 [M+Na] + .

[0197] Example 33 Synthesis of 6-(1-chloroethyl)thiopheno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-4,8-dione

[0198] 6-(1-hydroxyethyl)thiopheno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-4,8-dione (1.0 g, 3.76 mmol) and dry DCM (20 mL) were added to a 100 mL three-necked flask. The mixture was cooled to 0 °C and thionyl chloride (0.54, 4.51 mmol) was added dropwise. The mixture was then allowed to react at room temperature. After 4 h, the reaction was confirmed to be complete by TLC. 10 mL of water was added to the reaction mixture, and the mixture was washed once with DCM (20 mL × 3) and saturated saline (30 mL). The mixture was dried over anhydrous magnesium sulfate for 4 h, filtered, and concentrated to obtain a yellow solid (0.88 g, 82.0%). 1H NMR (400MHz, CDCl3): δ (ppm) 7.74 (d, J = 0.8Hz, 1H), 5.38-5.33 (m, 1H), 2.0 (d, J = 6.8Hz, 3H), ESI-MS: m / z 306.9 [M+Na] + .

[0199] Example 34 Synthesis of 6-(1-(4-methylpiperazin-1-yl)ethyl)thiopheno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-4,8-dione (SD-1)

[0200] Add 0.15 g (0.53 mmol) of 6-(1-chloroethyl)thiopheno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-4,8-dione, 5 mL of dry acetonitrile, 0.15 g (1.06 mmol) of K2CO3, 9 mg (0.053 mmol) of KI, and 64 mg (0.64 mmol) of N-methylpiperazine to a 25 mL single-necked flask. Protect with Ar and heat to 60 °C. After 4 h, the reaction was detected by TLC to indicate that the reaction was complete. Remove the acetonitrile under reduced pressure, wash once with DCM (10 mL × 3) and saturated saline (30 mL), dry with anhydrous magnesium sulfate for 4 h, filter, concentrate, and obtain 0.18 g of yellow solid SD-1. Mix with 0.25 g of silica gel, pack 1 g of silica gel into a column, and use EA:PE = 1:1 as the mobile phase to obtain SD-1 (65 mg, 36.11%). mp184-186℃. 1 H NMR (400MHz, CDCl3): δ (ppm) 7.56 (d, J = 1.0Hz, 1H), 4.01-3.96 (m, 1H), 2.70-2.64 (m, 8H), 2.54 (s, 3H), 1.48 (d, J = 6.8Hz, 3H), 13 C NMR (100MHz, CDCl3): δ172.23,170.78,164.55,156.88,156.73,145.49,143.75,122.84,54.69,55.14,45.82,15.42.

[0201] Example 35 Synthesis of 6-(1-(4-(2-hydroxyethyl)piperazin-1-yl)ethyl)thiopheno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-4,8-dione (SD-2)

[0202] Following the method described in Example 34, a white solid, SD-2, was obtained from 2-(piperazin-1-yl)ethane-1-ol as a raw material. The yield was 60 mg, the recovery rate was 30%, and the mp value was 190-192 °C. 1 H NMR (400MHz, CDCl3): δ (ppm) 7.56 (s, 1H), 4.02-3.97 (m, 1H), 3.64 (t, J = 4.8Hz, 2H), 2.68-2.60 (m, 12H), 1.49 (d, J = 6.6Hz, 3H). 13 C NMR (100MHz, CDCl3): δ172.22,170.78,164.29,156.88,156.74,145.50,143.75,122.89,59.70,59.24,57.67,53.01,15.52.

[0203] Example 36 Synthesis of 6-(1-(4-(pyrimidin-2-yl)piperazin-1-yl)ethyl)thiopheno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-4,8-dione (SD-3)

[0204] Following the method described in Example 34, a white solid, SD-3, was obtained from 2-(piperazine-1-yl)pyrimidine as a raw material, with a yield of 70 mg and a recovery rate of 32%, at mp 195-197°C. 1 H NMR (400MHz, CDCl3): δ (ppm) 8.30 (d, J = 4.7Hz, 2H), 7.59 (s, 1H), 6.49 (t, J = 4.7H z,1H),4.06-4.05(m,1H),3.89(s,4H),2.71-2.64(m,4H),1.51(d,J=6.1Hz,3H). 13 C NMR (100MHz, CDCl3): δ172.20,170.22,163.94,161.52,157.73,156.86,156.72,145.63,143.72,123.04,110.03,59.98,49.18,43.84,15.41.

[0205] Example 37 Synthesis of (3S)-1-(1-(4,8-dioxo-4,8-dihydrothiopheno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazol-6-yl)ethyl)piperidine-3-carboxylic acid ethyl ester (SD-4)

[0206] Following the method described in Example 34, a white solid, SD-4, was obtained from (S)-piperidine-3-carboxylic acid ethyl ester as a raw material. The yield was 77 mg, the recovery rate was 36%, and the mp was 191-193 °C. 1 H NMR (400MHz, CDCl3): δ7.55(s,1H),.4.19-4.10(m,2H),4.09-4.02(m,1H),3.04-2.86(m,1H),2.66(s,1H),2.62-2 .57(m,1H),2.45(s,1H),2.24-2.20(m,1H),1.97(s,1H),1.78(s,1H),1.62(s,2H),1.49(s,3H),1.27-1.22(m,3H). 13 C NMR (100MHz, CDCl3): δ173.84,172.23,170.74,164.73,156.88,156.73,145.58, 143.73,122.67,60.50,60.17,53.42,51.12,42.19,41.91,26.87,24.83,14.22.

[0207] Example 38 Synthesis of 1-(1-(4,8-dioxo-4,8-dihydrothiopheno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazol-6-yl)ethyl)piperidine-4-carboxylic acid ethyl ester (SD-5)

[0208] Following the method described in Example 34, a white solid, SD-5, was obtained from piperidine-4-carboxylic acid ethyl ester as a raw material, with a yield of 75 mg and a recovery rate of 35%, at mp 197-199 °C. 1 H NMR (400MHz, CDCl3): δ7.55 (s, 1H), 4.14 (q, J = 7.2Hz, 2H), 4.03-3.98 (m, 1H), 2.91-2.88 (m, 1H), 2.82-2.79 (m, 1H), 2. 46-2.41(m,1H),2.33-2.21(m,2H),1.94-1.92(m,2H),1.87-1.75(m,2H),1.46(d,J=6.9Hz,3H),1.27(t,J=7.1Hz,3H). 13C NMR (100MHz, CDCl3): δ174.77,172.27,170.74,156.91,156.75,145.53,143.72, 129.62,122.61,60.42,59.97,50.46,47.31,41.01,28.59,28.31,14.05,14.23.

[0209] Example 39 Synthesis of 6-(1-(bis(2-hydroxyethyl)amino)ethyl)thiopheno[2',3':4,5]benzo[1,2-c][1,2,5]thiadiazole-4,8-dione (SD-6)

[0210] Following the method described in Example 34, a white solid, namely SD-6, was obtained from diethanolamine as a raw material, with a yield of 68 mg and a recovery rate of 37%, at mp 193-195°C. 1 H NMR (400MHz, CDCl3): δ7.60 (d, J = 1.0Hz, 1H), 4.28 (q, J = 6.4Hz, 1H), 3.80-3.74 (m, 2H), 3.72-3.67 (m, 2H), 2.78-2.75 (m, 6H), 1.57 (d, J = 6.7Hz, 3H). 13 C NMR (100MHz, CDCl3): δ172.15,170.63,163.18,156.78,156.67,145.60,143.55,123.67,60.54,56.84,51.90,19.68.

[0211] Example 40 Synthesis of methyl 5,9-dimethoxythiopheno[2,3-g]quinoxaline-7-carboxylate

[0212] Add 50 mL of THF to a 500 mL single-necked flask containing methyl 5,6-diamino-4,7-dimethoxybenzo[b]thiophene-2-carboxylate (3.74 g, 13.26 mmol). Heat to 60 °C, add glyoxal (3.85 g, 26.52 mmol). After 0.5 h, the reaction is confirmed by TLC to be complete. Evaporate the THF under reduced pressure, pour the residue into 20 mL of water, and extract three times with 30 mL of DCM. Combine the organic phases, wash once with 20 mL of saturated NaCl, dry with anhydrous magnesium sulfate for 4 h, filter, concentrate, and obtain 4.14 g of crude product. Mix with 4.97 g of silica gel, pack a 40 g silica gel column, and use EA:PE = 1:5 as the mobile phase to obtain the product (2.4 g, 60%). 1H NMR (400MHz, CDCl3): δ8.84(d,J=1.4,1H,), 8.79(d,J=1.4,1H), 8.34(s,1H), 4.29(s,3H), 4.27(s,3H), 3.93(s,3H).ESI-MS: m / z 305.0[M+H] + 327.0 [M+Na] + , 631.1[2M+Na] + .

[0213] Example 41 Synthesis of 5,9-dimethoxythiopheno[2,3-g]quinoxaline-7-carboxylic acid

[0214] Add 50 mL of THF containing 3.35 g (11.02 mmol) of 5,9-dimethoxythiophene[2,3-g]quinoxaline-7-carboxylic acid methyl ester (THF) to a 250 mL single-necked flask, followed by 0.92 g (22.04 mmol) of NaOH. After this, slowly add 50 mL of H₂O. After the addition is complete, the reaction is monitored by TLC after 30 min to ensure completeness. Adjust the pH to 1-2 with 1 N HCl under ice bath conditions, evaporate the THF under reduced pressure, pour the solution into 30 mL of ice water, and filter to obtain (2.63 g, 82.19%). ESI-MS: m / z 291.00 [M+H] + 290.0 [MH] - .

[0215] Example 42 Synthesis of 5,9-dioxo-5,9-dihydrothiopheno[2,3-g]quinoxaloline-7-carboxylic acid

[0216] Add 0.5 g (1.72 mmol) of 5,9-dimethoxythieno[2,3-g]quinoxaloline-7-carboxylic acid and 10 mL of acetonitrile to a 50 mL three-necked flask. Cool to 0 °C, and add 10 mL of H₂O containing 1.86 g (3.96 mmol). After addition, the reaction is confirmed to be complete by TLC. Quench the solution with 20 mL of ice water, filter, and obtain a yellow solid (0.39 g, 88.64%). ESI-MS: m / z 261.0 [M+H] + 283.0 [M+Na] + .

[0217] Example 43 Synthesis of N,N-diethyl-5,9-dioxo-5,9-dihydrothiophene[2,3-g]quinoxaline-7-carboxamide (SG-1)

[0218] Add 5,9-dioxo-5,9-dihydrothiopheno[2,3-g]quinoxaloline-7-carboxylic acid (0.20 g, 0.77 mmol), dry DCM (5 mL), HATU (0.35 g, 0.92 mmol), and DIPEA (0.24 g, 1.8 mmol) to a 100 mL single-necked flask. After 0.5 h, add diethylamine (67 mg, 0.92 mmol) dropwise. After 1 h, the reaction is confirmed to be complete by TLC. Remove the solvent under reduced pressure and add 1... 0 mL of water was added to 10 mL of water. The solution was washed once with DCM (10 mL × 3), 1 N HCl (10 mL × 2), saturated NaCO3 (10 mL × 1), and saturated saline (20 mL). The solution was dried over anhydrous magnesium sulfate for 4 h, filtered, and concentrated to obtain 0.36 g of pale yellow solid SG-1. The solution was mixed with 0.43 g of silica gel and packed into a column with 1 g of silica gel. The mobile phase was EA:PE = 2:1 to obtain SG-1 (82 mg, 34%), mp 184-187℃. 1 H NMR (400MHz, CDCl3): δ (ppm) 9.08 (s, 2H), 7.89 (s, 1H), 3.57 (m, 4H), 1.31 (t, 6H), 13 C NMR (100MHz, CDCl3): δ176.55,175.30,161.37,148.64,148.61,148.43,145. 67,145.31,141.74,125.72,77.37,77.05,76.73,43.79,41.32,14.47,12.84.

[0219] Example 44 Synthesis of (S)-1-(5,9-dioxo-5,9-dihydrothieno[2,3-g]quinoxaloline-7-carbonyl)piperidine-3-carboxylic acid ethyl ester (SG-2)

[0220] Following the method described in Example 43, a white solid, SG-2, was obtained from (S)-piperidine-3-carboxylic acid ethyl ester as a raw material. The yield was 98 mg, the recovery rate was 34%, and the mp was 189-191 °C. 1 H NMR (400MHz, CDCl3): δ9.08(m,2H),7.90(s,1H),4.23-4.12(m,3H),4.02-3.99(m,1H),3.42-3.36(m,1 H),2.65-2.60(m,1H),2.16-2.14(m,1H),1.89-1.86(m,2H),1.67-1.63(m,2H),1.27(t,J=7.1Hz,3H). 13C NMR (100MHz, CDCl3): δ176.52,175.25,172.44,161.22,148.64,148.46,14 5.92,145.61,145.28,141.55,126.37,61.03,41.29,27.14,24.42,14.19.

[0221] Example 45 Synthesis of (R)-1-(5,9-dioxo-5,9-dihydrothiopheno[2,3-g]quinoxaloline-7-carbonyl)piperidine-3-carboxylic acid ethyl ester (SG-3)

[0222] Following the method described in Example 43, a white solid, SG-3, was obtained from (R)-piperidine-3-carboxylic acid ethyl ester as a raw material, with a yield of 95 mg and a recovery rate of 31%, at mp 186-188°C. 1 H NMR (400MHz, CDCl3): δ9.08(s,2H),7.90(s,1H),4.24-4.14(m,3H),4.02-3.99(m,1H),3.41-3.36(m,1 H),2.64-2.60(m,1H),2.16-2.14(m,1H),1.89-1.86(m,2H),1.67-1.63(m,2H),1.27(t,J=7.1Hz,3H). 13 C NMR (100MHz, CDCl3): δ176.52,175.25,172.44,161.22,148.65,148.46,145.93,145.61,145.28,141.55,126.37,61.04,41.29,27.14,14.19.

[0223] Example 46 Synthesis of 7-(piperidine-1-carbonyl)thieno[2,3-g]quinoxaline-5,9-dione (SG-4)

[0224] Following the method described in Example 43, a white solid, SG-4, was obtained from piperidine as a raw material, with a yield of 81 mg and a recovery rate of 32%, at mp 187-189°C. 1 H NMR (400MHz, CDCl3): δ9.08-9.07(m,2H),7.85(s,1H),3.69(s,4H),1.78-1.70(m,6H). 13C NMR (100MHz, CDCl3): δ176.56,175.25,160.75,148.60,148.43,148.08,145.73,145.65,145.30,141.58,126.11,25.77,24.36.

[0225] Example 47 Synthesis of tert-butyl 2-(5,9-dioxo-5,9-dihydrothieno[2,3-g]quinoxaloline-7-carboxamido)ethyl)carbamate (SG-5)

[0226] Following the method described in Example 43, a white solid, SG-5, was obtained from NB℃ ethylenediamine as a raw material, with a yield of 93 mg and a recovery rate of 30%, at mp 188-190℃. 1 H NMR (400MHz, CDCl3): δ9.11-9.02(m,2H),8.37(brs,1H),8.34(s,1H),6.94(t,J=5.4Hz,3H),3.34-3.16(m,2H),3.14-3.13(m,2H),1.38(m,9H). 13 C NMR (100MHz, CDCl3): δ182.19,181.22,164.98,160.93,153.42,153.37,153.13,157.68,151.01,150.75,147.68,131.01,82.98,33.45,25.98.

[0227] Example 48 Synthesis of 5,9-dioxo-N-(2-(piperidin-1-yl)ethyl)-5,9-dihydrothiopheno[2,3-g]quinoxaline-7-carboxamide (SG-6)

[0228] Following the method described in Example 43, a white solid, SG-6, was obtained from 2-(piperidin-1-yl)ethane-1-amine as a raw material. The yield was 94.0 g, with a recovery rate of 33%, and the mp value was 191-193 °C. 1 H NMR (400MHz, DMSO-d6): δ (ppm) 9.58 (s, 1H), 9.11 (q, J = 2.28Hz, 2H), 8.52 (s, 2H), 3.71 (d, J = 5. 60Hz,2H),3.53(s,2H),3.25(s,2H),2.95-2.89(m,2H),1.80(s,2H),1.70(s,1H),1.41(s,1H) 13C NMR (100MHz, DMSO-d6): δ177.36,176.49,160.57,148.66,148.41,147.88,147.19, 146.23,146.01,142.93,126.79,55.39,52.72,49.06,41.74,34.76,22.85,21.79.

[0229] Example 49 Synthesis of 5,9-dioxo-N-(2-(pyrrolidone-1-yl)ethyl)-5,9-dihydrothiopheno[2,3-g]quinoxaline-7-carboxamide (SG-7)

[0230] Following the method described in Example 43, a white solid, SG-7, was obtained from 2-(pyrrolidone-1-yl)ethane-1-amine as a raw material. The yield was 93 mg, the recovery rate was 34%, and the mp value was 190-192 °C. 1 H NMR (400MHz, DMSO-d6): δ (ppm) 9.43 (s, 1H), 9.11 (s, 2H), 8.50 (s, 1H), 3.62 (d, J = 3.24Hz, 2H), 3.20-3.17 (m, 6H), 1.90 (s, 4H), 13 C NMR (100MHz, DMSO-d6): δ177.37,176.48,160.51,148.66,148.40,148.11,147.10,146.22,146.00,142.39,126.72,55.39,53.75x 2,37.07,23.19.

[0231] Example 50 Synthesis of N-(2-morpholinoethyl)-5,9-dioxo-5,9-dihydrothiopheno[2,3-g]quinoxaline-7-carboxamide (SG-8)

[0232] Following the method described in Example 43, a white solid, SG-8, was obtained from 2-morpholinoethanol-1-amine as a raw material. The yield was 89 mg, the recovery rate was 31%, and the mp value was 194-196 °C. 1 H NMR (400MHz, DMSO-d6): δ (ppm) 9.58 (s, 1H), 9.11 (q, J = 2.28Hz, 2H), 8.52 (s, 2H), 3.71 (d, J = 5. 60Hz,2H),3.53(s,2H),3.25(s,2H),2.95-2.89(m,2H),1.80(s,2H),1.70(s,1H),1.41(s,1H) 13C NMR (100MHz, DMSO-d6): δ177.36,176.49,160.57,148.66,148.41,147.88,147.19, 146.23,146.01,142.93,126.79,55.39,52.72,49.06,41.74,34.76,22.85,21.79.

[0233] Example 51 Synthesis of (S)-(1-(5,9-dioxy-5,9-dihydrothieno[2,3-g]quinoxaline-7-carbonyl)piperidin-3-yl)tert-butyl carbamate (SG-12)

[0234] Following the method described in Example 43, a white solid, SG-12, was obtained from (S)-piperidin-3-ylcarbamate tert-butyl ester as a raw material. The yield was 105 mg, the recovery rate was 31%, and the mp was 185-187 °C. 1 H NMR (400MHz, CDCl3): δ (ppm) 9.07 (d, J = 1.29Hz, 2H), 7.98 (s, 1H), 4.65 (s, 1H), 4.00 (s, 1H), 3.75 ( s,2H),3.64(s,1H),3.49(s,1H),2.04-2.01(m,1H),1.84-1.83(m,1H),1.70(s,2H),1.42(s,9H), 13 C NMR (100MHz, CDCl3): δ176.39,175.26,161.62,155.03,148.63,148.40,147.48,145. 99,145.58,145.30,141.66,126.80,80.18,46.94,31.24,29.97,29.70,28.35,22.69.

[0235] Example 52 Synthesis of (R)-(1-(5,9-dioxy-5,9-dihydrothieno[2,3-g]quinoxaline-7-carbonyl)piperidin-3-yl)tert-butyl carbamate (SG-13)

[0236] Following the method described in Example 43, a white solid, SG-13, was obtained from (R)-piperidin-3-ylcarbamate tert-butyl ester as a raw material. The yield was 100 mg, the recovery rate was 29%, and the mp was 188-190 °C. 1H NMR (400MHz, CDCl3): δ (ppm) 8.07 (d, J = 2.00Hz), 7.98 (s, 1H), 4.65 (s, 1H), 4.00 (s, 1H), 3.75 (s, 2H) ),3.64(s,1H),3.49(s,1H),2.04-2.00(m,1H),1.83-1.73(m,1H),1.72-1.68(m,2H),1.42(s,2H), 13 C NMR (100MHz, CDCl3): δ176.40,175.26,161.63,155.03,148.63,148.40,147.48,145. 99,145.59,145.31,141.66,126.80,80.18,46.94,31.24,29.96,29.70,28.36,22.69.

[0237] Example 53 Synthesis of 4-(5,9-dioxo-5,9-dihydrothieno[2,3-g]quinoxaline-7-carboxamido)piperidine-1-carboxylic acid tert-butyl ester (SG-14)

[0238] Following the method described in Example 43, a white solid, SG-14, was obtained from tert-butyl 4-aminopiperidine-1-carboxylate as a raw material. The yield was 102 mg, the recovery rate was 30%, and the mp value was 189-191 °C. 1 H NMR (400MHz, DMSO-d6): δ (ppm) 9.10 (d, J = 1.00Hz, 2H), 8.84 (d, J = 7.68Hz, 1H), 8.46 ( s,1H),3.96-3.93(m,3H),2.87(s,2H),1.85-1.82(m,2H),1.47(s,2H),1.42(s,9H), 13 C NMR (100MHz, DMSO-d6): δ177.45,159.28,154.41,148.70,146.64,148.40,1 47.10,146.24,145.97,142.98,126.26,79.19,47.50,42.48,31.56,28.56.

[0239] Example 54 Synthesis of 7-(4-(bis(4-fluorophenyl)methyl)piperazine-1-carbonyl)thieno[2,3-g]quinoxaline-5,9-dione (SG-15)

[0240] Following the method described in Example 43, a white solid, SG-15, was obtained from 1-(bis(4-fluorophenyl)methyl)piperazine as a raw material. The yield was 106 mg, the recovery rate was 26%, and the mp was 225-227 °C. 1 HNMR (400MHz, CDCl3): δ (ppm) 9.06 (s, 2H), 7.82 (s, 1H), 7.36 (s, 4H), 7.00 (s, 4H), 4.31 (s, 1H), 3.77 (s, 4H), 2.48 (s, 4H) 13 CNMR (100MHz, CDCl3): δ176.46,175.19,163.26,160.81,160.75,148.65,148.47,147.35,145.97,14 5.59,145.30,141.48,137.26,137.23,129.28,129.20,126.46,115.82,115.61,74.19,51.62,29.70.

[0241] Example 55 Synthesis of 7-(4-benzylpiperazine-1-carbonyl)thieno[2,3-g]quinoxaline-5,9-dione (SG-16)

[0242] Following the method described in Example 43, a white solid, SG-16, was obtained from 1-benzylpiperazine as a raw material, with a yield of 95 mg and a recovery rate of 25%, at mp 213-215°C. 1 H NMR (400MHz, CDCl3): δ (ppm) 9.06 (s, 2H), 7.82 (s, 1H), 7.42 (d, J = 7.32, 4H), 7.30 (t, J = 7.28Hz, 6H), 4.31 (s, 1H), 3.78 (s, 4H), 2.51 (s, 4H) 13 C NMR (100MHz, CDCl3): δ176.46,175.20,160.72,148.62,148.44,147.54,145.92,145 .61,145.25,141.75,141.49,128.73,128.41,127.86,127.36,126.42,75.84,51.78.

[0243] Example 56 Synthesis of 4-(5,9-dioxo-5,9-dihydrothiopheno[2,3-g]quinoxaline-7-carbonyl)piperazine-1-carboxylic acid tert-butyl ester (SG-18)

[0244] Following the method described in Example 43, a white solid, SG-18, was obtained from piperazine-1-carboxylic acid tert-butyl ester as a raw material. The yield was 79 mg, the recovery rate was 24%, and the mp was 181-183 °C. 1 HNMR (400MHz, CDCl3): δ (ppm) 9.08 (d, J = 0.76Hz, 2H), 7.81 (s, 1H), 3.76-3.74 (m, 4H), 3.57-3.55 (m, 4H), 1.49 (s, 9H).

[0245] Example 57 Synthesis of 7-(4-benzylpiperazine-1-carbonyl)thieno[2,3-g]quinoxaline-5,9-dione (SG-19)

[0246] Following the method described in Example 43, a white solid, SG-19, was obtained from 1-benzylpiperazine as a raw material, with a yield of 90 mg and a recovery rate of 28%, at mp 182-184 °C. 1 H NMR (400MHz, CDCl3): δ (ppm) 9.07 (s, 2H), 7.84 (s, 1H), 7.32-7.15 (m, 5H), 4.61 (s, 1H), 4.1 1(s,1H),3.09-2.92(m,2H),2.61(d,J=6.96Hz,2H),1.83-1.79(m,2H),1.37-1.29(m,2H).

[0247] Example 58 Synthesis of 7-(4-benzylpiperazine-1-carbonyl)thieno[2,3-g]quinoxaline-5,9-dione (SG-22)

[0248] Following the method described in Example 43, a white solid, SG-22, was obtained from 1-methylpiperazine as a raw material. The yield was 72 mg, the recovery rate was 23%, and the mp value was 187-189 °C. 1 H NMR (400MHz, DMSO-d6): δ (ppm) 9.10 (s, 2H), 7.94 (s, 1H), 3.66 (t, J = 4.68Hz, 4H), 2.43 (t, J = 4.48Hz, 4H), 2.24 (s, 3H). 13 C NMR (100MHz, CDCl3): δ177.21,176.26,160.90,150.13,148.69,148.42,146.12,146.00,145.93,142.09,126.90,54.65,45.86.

[0249] Example 59 Synthesis of N-benzyl-5,9-dioxo-5,9-dihydrothiopheno[2,3-g]quinoxaline-7-carboxamide (SG-23)

[0250] Following the method described in Example 43, a white solid, SG-23, was obtained from diphenylmethylamine as a raw material. The yield was 68 mg, the recovery rate was 20%, and the mp value was 192-194 °C. 1 HNMR (400MHz, DMSO-d6): δ (ppm) 9.83 (d, J = 8.44Hz, 1H), 9.10 (s, 2H), 8.71 (s, 1H), 7.39-7.38 (m, 8H), 7.33-7.28 (m, 2H), 6.38 (d, J = 8.4Hz, 1H). 13 C NMR (100MHz, DMSO-d6): δ177.43,176.46,159.59,148.65,148.41,148.11,147 .33,146.25,145.99,143.04,141.88,128.95,128.13,127.78,127.04,57.34.

[0251] Example 60 Synthesis of N,N-dibenzyl-5,9-dioxo-5-9-dihydrothiopheno[2,3-g]quinoxaline-7-carboxamide (SG-24)

[0252] Following the method described in Example 43, a white solid, SG-24, was obtained from dibenzylamine as a raw material, with a yield of 68 mg and a recovery rate of 20%, at mp 195-197°C. 1 H NMR (400MHz, CDCl3): δ (ppm) 9.05 (s, 2H), 7.89 (s, 1H), 7.42-7.25 (m, 10H), 4.71 (s, 4H). 13 C NMR (100MHz, CDCl3): δ176.36,175.22,162.87,148.63,148.42,147.61,1 46.07,145.57,145.25,141.67,135.42,129.15,128.20,127.02,126.46.

[0253] Test Example 1

[0254] 1. Cell line

[0255] Human hepatocellular carcinoma HepG2 cells were purchased from the cell bank of the Shanghai Institute of Cell Biology, Chinese Academy of Sciences.

[0256] 2. Experimental Materials

[0257] Cell Counting Kit-8 (CCK-8) (Beyotime, Shanghai, China), 96-well cell culture plates (Costar), 10% fetal bovine serum (TBD Science, Tianjin, China), penicillin / streptomycin (Sigma, St. Louis, MO, USA), culture medium (gibco, NY, USA), microplate reader (PerkinElmer EnSpire).

[0258] 3. Experimental Procedure

[0259] Cell culture and cell viability assay

[0260] Human liver cancer HepG2 cells were cultured in Dulbecco modified medium containing high concentrations of glucose, supplemented with 10% fetal bovine serum and penicillin / streptomycin, and placed in a 37°C, 5% CO2 incubator. Cells were passaged every 3-4 days, and cells in the logarithmic growth phase were selected for experiments.

[0261] HepG2 cells were cultured at 4 × 10⁻⁸ using Cell Counting Kit-8 (CCK-8). 3 K562 cells were seeded at a density of 4 × 10⁶ cells / well in 96-well plates and grown for 24 h. 3 Cells were seeded at a density of 10 cells / well in 96-well plates. After adding different concentrations of the test compound, the cells were cultured for 72 hours. Then, 10 μl of CCK-8 reagent was added to each well, and the culture plate was incubated at 37°C for 4 hours. The optical density (OD) at 530 / 590 nm was measured using a microplate reader. Cell viability was expressed as a percentage of the untreated control. Each treatment was performed in triplicate.

[0262] The method for calculating the inhibition rate of tumor cell growth by the test compound is as follows:

[0263] Tumor cell growth inhibition rate % = 1 - (OD experiment - OD blank) / (OD control - OD blank).

[0264] in:

[0265] The OD experiment represents the absorbance value of the sample well (cells + analyte compound + CCK-8);

[0266] OD control indicates the absorbance of the control well (cells + DMSO + CCK-8);

[0267] OD blank represents the absorbance value of the zeroing well (culture medium + DMSO + CCK-8);

[0268] OD control = OD570 control - OD630 control;

[0269] OD blank = OD570 blank - OD630 blank.

[0270] Fitting of the inhibition curve of the tested compound on tumor cell growth and IC50. 50 Calculation;

[0271] 4. Experimental Results

[0272] Table 1. Inhibitory activity of SC, SD and SG series compounds against HepG2

[0273]

[0274] As shown in Table 1, the SC, SD, and SG series compounds provided by this invention are effective against HepG2 IC. 50 The values ​​ranged from 0.08 nM to 4.81 μM, exhibiting excellent inhibitory activity. Experimental results showed that SC-7 exhibited the strongest inhibitory activity against HepG2, with an IC50 value of [missing value]. 50 The concentration was 0.08 μM. Experimental results showed that all compounds exhibited superior inhibitory activity against HepG2 compared to the lead compound BBI608, demonstrating excellent development potential.

[0275] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A BBI608-type derivative, characterized in that, Having the structure shown in SD-1, SD-2, SD-3, SD-4, SD-5, SD-6, SC-1, SC-2, SC-3, SC-4, SC-5, SC-6, SC-7, SC-8, SC-9, SC-11, SC-15, SC-16, SC-17, SC-23, SC-24 or SC-27:

2. The method for preparing the BBI608 derivative of claim 1, comprising the following steps: (i) When the BBI608 derivative has the structure shown in SC-1, SC-2, SC-3, SC-4, SC-5, SC-6, SC-7, SC-8, SC-9, SC-11, SC-15, SC-16, SC-17, SC-23, SC-24 or SC-27, the preparation method includes the following steps: Compound a undergoes a first nucleophilic substitution reaction with a nitrating agent to give compound b; Compound b and compound 1 undergo a first cladding reaction to yield compound c; Compound c undergoes a second nucleophilic substitution reaction with a nitrating agent to give compound d; The compound d undergoes a first reduction reaction to yield compound e; Compound e and compound 2 undergo a second ring-cladding reaction to yield compound f; The compound f undergoes a first hydrolysis reaction to yield compound g; The compound g undergoes a first oxidation reaction to yield compound h; In the presence of 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate and N,N-diisopropylethylamine, compound h undergoes a first acylation reaction with compound 3 to give BBI608 derivatives. PhNOS compound 2; R1H compound 3; (ii) When the BBI608 derivative has the structure shown in SD-1, SD-2, SD-3, SD-4, SD-5 or SD-6, the preparation method includes the following steps: Compound g and compound 4 undergo a second acylation reaction to yield compound i; Compound i undergoes a first electrophilic substitution reaction with a Grignard reagent to obtain compound j; Compound j undergoes a second reduction reaction to yield compound k; The compound k undergoes a second oxidation reaction to yield compound l; Compound l undergoes a third nucleophilic substitution reaction with a chlorinating reagent to give compound m; Compound m undergoes a fourth nucleophilic substitution reaction with compound 3 to obtain BBI608-type derivatives; R1H compound 3; 3. The use of the BBI608 derivative of claim 1 or the BBI608 derivative prepared by the preparation method of claim 2 in the preparation of drugs for treating STAT3-mediated diseases; wherein the STAT3-mediated diseases are one or more of colorectal cancer, lung cancer, melanoma, breast cancer, prostate cancer, kidney cancer, ovarian cancer, liver cancer, pancreatic cancer, multiple myeloma, and leukemia.

4. A medicament for treating STAT3-mediated diseases, the active ingredient comprising the BBI608 derivative of claim 1 and a pharmaceutically acceptable salt thereof.

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