CN117567361A - Aryl alkyne compound as well as preparation method and application thereof - Google Patents

Aryl alkyne compound as well as preparation method and application thereof Download PDF

Info

Publication number
CN117567361A
CN117567361A CN202311519441.8A CN202311519441A CN117567361A CN 117567361 A CN117567361 A CN 117567361A CN 202311519441 A CN202311519441 A CN 202311519441A CN 117567361 A CN117567361 A CN 117567361A
Authority
CN
China
Prior art keywords
compound
acid
cancer
stirring
drying
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN202311519441.8A
Other languages
Chinese (zh)
Inventor
徐学军
徐春萍
杨玉坡
杨争艳
徐红运
段超群
裴梦富
刘亚青
汤善顺
李岑
傅凯
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Henan Radiomedical Science And Technology Co ltd
Original Assignee
Henan Radiomedical Science And Technology Co ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Henan Radiomedical Science And Technology Co ltd filed Critical Henan Radiomedical Science And Technology Co ltd
Priority to CN202311519441.8A priority Critical patent/CN117567361A/en
Publication of CN117567361A publication Critical patent/CN117567361A/en
Pending legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
    • C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
    • C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D213/24—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with substituted hydrocarbon radicals attached to ring carbon atoms
    • C07D213/54—Radicals substituted by carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
    • C07D213/56—Amides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/02—Antineoplastic agents specific for leukemia
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
    • C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
    • C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D213/60—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D213/78—Carbon atoms having three bonds to hetero atoms, with at the most one bond to halogen, e.g. ester or nitrile radicals
    • C07D213/81—Amides; Imides
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
    • C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
    • C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D213/60—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D213/78—Carbon atoms having three bonds to hetero atoms, with at the most one bond to halogen, e.g. ester or nitrile radicals
    • C07D213/81—Amides; Imides
    • C07D213/82—Amides; Imides in position 3
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D215/00—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems
    • C07D215/02—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom
    • C07D215/16—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D215/48—Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D217/00—Heterocyclic compounds containing isoquinoline or hydrogenated isoquinoline ring systems
    • C07D217/22—Heterocyclic compounds containing isoquinoline or hydrogenated isoquinoline ring systems with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to carbon atoms of the nitrogen-containing ring
    • C07D217/26—Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D241/00—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings
    • C07D241/02—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings not condensed with other rings
    • C07D241/10—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings not condensed with other rings having three double bonds between ring members or between ring members and non-ring members
    • C07D241/14—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings not condensed with other rings having three double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D241/24—Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/06—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a carbon chain containing only aliphatic carbon atoms
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00—Technologies relating to chemical industry
    • Y02P20/50—Improvements relating to the production of bulk chemicals
    • Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Hematology (AREA)
  • Oncology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

The invention discloses an aryl alkyne compound, a preparation method and application thereof, wherein the structural formula is shown in a general formula I:wherein the A ring is selected from The B ring is selected from

Description

Aryl alkyne compound as well as preparation method and application thereof
Technical Field
The invention belongs to the field of tumor targeted therapy, and particularly relates to an aryl alkyne compound, a preparation method and application thereof.
Background
Signal transducers and transcriptional activators (Signal transducer and activators of transcriptions, STATs) are a family of seven transcription factors (STAT 1, STAT2, STAT3, STAT4, STAT5, STAT6 and STAT 7) that regulate expression of key proteins encoding the entire proteome and thus are involved in regulating cellular metabolism, intercellular communication, immune response and cell cycle. Activation of STATs is typically initiated by ligand binding of a cell surface receptor, followed by kinase-dependent phosphorylation of tyrosine (pY) residues. This results in Src 2 homology (SH 2) domain mediated STAT-STAT dimerization, rapid translocation to the nucleus, binding to DNA elements and initiation of transcription of downstream genes. Recent studies have shown that STATs can not only signal transcription, but also regulate mitochondrial anabolism and catabolism and can affect functional partitioning of the nucleus and genome integrity. Studies have reported that mutation of the function of STATs proteins (i.e. somatic or desired variation) is the basis for tumor cell carcinogenesis. In human lung cancer, breast cancer, prostate cancer, colorectal cancer, liver cancer, blood tumor and the like, there are driving mutations of STAT3 pathway, resulting in abnormal enhancement of STAT3 activity, and are closely related to malignant progress and bad survival prognosis of tumor. Targeting STAT3 is considered a well-promising tumor treatment strategy. Currently, only three JAKs inhibitors are internationally used for treating immune diseases, and the application of JAK/STATs signal inhibitors in tumors is still in a research and development stage.
In order to develop STAT3 targeted antitumor drugs, an aryl alkyne compound with a brand new structural formula is recently synthesized. Through analysis of some biological technologies, the compounds can obviously inhibit phosphorylation and activation of STAT3 signals, down regulate the expression of key proteins related to cell cycle and apoptosis process, inhibit the proliferation of various tumor cell lines such as gastric cancer, pancreatic cancer, breast cancer and the like, and show extremely strong tumor inhibition activity.
The invention aims to disclose an anti-tumor effect and a potential pharmacological mechanism of aryl alkyne compounds and derivatives thereof, and potential application of the compounds in clinical treatment of gastric cancer, pancreatic cancer, breast cancer, liver cancer, lung cancer, colon cancer, leukemia, lymphoma and multiple myeloma.
Disclosure of Invention
The invention aims to provide aryl alkyne compounds, and a preparation method and application thereof.
Based on the above purpose, the invention adopts the following technical scheme:
an aryl alkyne compound with a structural formula shown in a general formula I:
wherein the A ring is selected from
The B ring is selected from
The aryl alkyne compound is specifically a compound with the following structure:
the above arylalkyne compound can be a biologically acceptable salt with at least one of acetic acid, dihydrofolic acid, benzoic acid, citric acid, sorbic acid, propionic acid, oxalic acid, fumaric acid, maleic acid, hydrochloric acid, malic acid, phosphoric acid, sulfurous acid, sulfuric acid, vanillic acid, tartaric acid, ascorbic acid, boric acid, lactic acid and ethylenediamine tetraacetic acid.
The preparation method of the aryl alkyne compound comprises the following synthetic routes:
the synthesis steps are as follows
(1) Dissolving the compound 1, the compound 2, DIEA and HBTU in DMF, stirring at room temperature to react completely, pouring the reaction solution into water, precipitating, filtering, collecting solid and drying to obtain a compound 3;
(2) Compound 3, compound 4, triethylamine and Pd (PPh 3 ) 2 Cl 2 Dissolving CuI in DMF, stirring at 70-90 ℃ for complete reaction, diluting the reaction solution with ethyl acetate, washing with saline solution, drying the organic phase, spin-drying, stirring, and passing through a column to obtain a compound 5;
(3) Dissolving the compound 5 in methanol, adding an ethyl acetate solution of hydrogen chloride, stirring at room temperature to react completely, and directly spin-drying the reaction solution to obtain a compound 6;
(4) Dissolving the compound 6 and potassium carbonate in DMF, adding the compound 7, stirring at 40-60 ℃ for complete reaction, diluting the reaction liquid with ethyl acetate, washing with saline solution, drying the organic phase, spin-drying, stirring, and passing through a column to obtain the compound shown in the formula I.
Preferably, in step (1), the molar ratio of compound 1, compound 2, DIEA and HBTU is 1:1:2 (1.0-1.5); in step (2), compound 3, compound 4, triethylamine and Pd (PPh 3 ) 2 Cl 2 And CuI in a molar ratio of 1:2:3:0.02:0.01; in the step (3), the molar ratio of the compound 5 to the hydrogen chloride is 1:10; in the step (4), the molar ratio of the compound 6 to the potassium carbonate to the compound 7 is 1 (2-4): 1.0-1.5.
The application of the aryl alkyne compound and the biologically acceptable salt thereof in preparing an anti-tumor drug, wherein the anti-tumor drug is a drug for treating tumors related to STAT3 signaling.
Preferably, the drug for treating tumors related to STAT3 signaling refers to drugs for treating gastric cancer, pancreatic cancer, breast cancer, liver cancer, lung cancer, colon cancer, leukemia, lymphoma and multiple myeloma.
It is another object of the present invention to provide a class of small molecule compounds with targeted antitumor activity.
The tumor may be specifically a STAT3 high-expression or constitutively activated tumor including, but not limited to, gastric cancer, pancreatic cancer, breast cancer, liver cancer, lung cancer, colon cancer, leukemia, lymphoma, multiple myeloma, and the like.
Specifically, the invention synthesizes aryl alkyne compounds RDn001, RDn002, RDn with brand new structure
003, RDn004, RDn005, RDn006, RDn007, RDn008, RDn009, RDn010, RDn011, RDn012, RDn013, RDn014 and RDn015. The proliferation inhibition effect of the compounds on tumor cells is detected by an MTT method, and the inhibition effect of the compounds on STAT3 signal transduction is proved by an immunoblotting method.
The results show that the compounds RDn001, RDn002, RDn003, RDn004, RDn005, RDn006, RDn of the invention
007, RDn008, RDn009, RDn010, RDn011, RDn012, RDn013, RDn014 and RDn015 can effectively inhibit proliferation of gastric cancer, pancreatic cancer, breast cancer, liver cancer, lung cancer, colon cancer, leukemia, lymphoma and multiple myeloma cells.
In summary, the present invention provides a novel aryl alkyne compound and its derivatives for use in tumor therapy and potential molecular mechanisms.
Drawings
FIG. 1 is a Western blot of RDn001 (0, 10, 30 and 100 nM) after 24h treatment of various tumor cell lines.
Detailed Description
In order to make the technical purpose, technical scheme and beneficial effect of the present invention more clear, the technical scheme of the present invention is further described below with reference to the accompanying drawings and specific embodiments.
In the process according to the invention for the synthesis of the compounds of the formula I, the various starting materials used for the reaction are preparable by the person skilled in the art according to the prior art, or can be prepared by methods known from the literature, or can be obtained commercially. The intermediates, raw materials, reagents, reaction conditions and the like used in the above reaction schemes may be appropriately changed according to the knowledge already known to those skilled in the art.
In the present inventionUnless otherwise indicated, wherein: (i) The temperature is expressed in degrees centigrade (DEG C), and the operation is performed in a room temperature environment; more specifically, the room temperature is 20-30 ℃; (ii) Drying the organic solvent by a common drying method, evaporating the solvent by a rotary evaporator under reduced pressure, wherein the bath temperature is not higher than 50 ℃; the volume ratio of the developing agent to the eluent is equal; (iii) the reaction process is followed by Thin Layer Chromatography (TLC); (iv) The final product has satisfactory proton nuclear magnetic resonance 1 H-NMR)。
EXAMPLE 1 Synthesis of RDn004 Compounds
The compound RDn004 is named (4- (4- (2, 2-trifluomethoxy) benzyl) piperazin-1-yl) (6- ((4- (trifluormethyl) phenyl) ethyl) pyridin-3-yl) methane,
the synthetic route is as follows:
step 1.Tert-butyl 4- (6-bromoninoyl) piperazine-1-carboxylate (intermediate 3)
Raw material 1D (1 g,4.95mmol,1.0 eq), raw material 2 (0.92 g,4.95mmol,1.0 eq), DIEA (1.28 g,9.90mmol,2.0 eq) and HBTU (2.26 g,5.94mmol,1.2 eq) were dissolved in 20ml dmf and stirred at room temperature for 2 hours; TLC (PE/ea=2/1, rf/product=0.25) showed that the starting material was reacted completely, with a new spot; the reaction solution was poured into 100mL of water, precipitated, filtered, and the solid was collected and dried to give intermediate 3D (1.60 g, 87.4%) as a white solid.
Step 2.Tert-butyl 4- (6- ((4- (trifluoromethyl) phenyl) ethyl) nicotinyl) piperazine-1-carboxylate (intermediate 5)
Intermediate 3D (1 g,2.70mmol,1.0 eq), starting material 4D (0.92 g,5.40mmol,2.0 eq), triethylamine (0.82 g,8.10mmol,3.0 eq), pd (PPh) 3 ) 2 Cl 2 (38 mg,0.054mmol,0.02 eq) and CuI (20 mg,0.108mmol,0.01 eq) were dissolved in 20 mM LDMF and reacted at 80℃with stirring for 12 hoursWhen in use; TLC (DCM/meoh=20/1, rf/product=0.35) showed that the starting material was reacted completely, with a new spot; the reaction was diluted with 100mL of ethyl acetate and washed 3 times with saturated brine (100 mL x 3), the organic phase was dried, spin-dried and passed through the column and rinsed with (DCM/meoh=100/1-20/1) to give intermediate 5D (620 mg, 50.2%) as an off-white solid.
Step 3.1- (4- (2- (4- (((4-phosphoraphtalen-1-yl) oxy) methyl) phenyl) ethyl) piperazin-1-yl) -2-phenylethan-1-one (intermediate 6)
Intermediate 5D (620 mg,1.35mmol,1.0 eq) was dissolved in 20mL of methanol, ethyl hydrogen chloride acetate solution (4 mol/L,3.37mL,13.5mmol,10.0 eq) was added, and the reaction was stirred at room temperature for 2 hours; TLC (DCM/meoh=20/1, rf/product=0.01) showed that the starting material was reacted completely, a new spot was generated; the reaction solution was directly dried to give intermediate 6D (284 mg, 99%) as a white solid.
Step 4. (4- (4- (2, 2-trifluomethoxy) benzyl) piperazin-1-yl) (6- ((4- (trifluormethyl) phenyl) ethyl) pyridin-3-yl) methane (RDn 004)
Intermediate 6D (400 mg,1.11mmol,1.0 eq) and potassium carbonate (463mg, 3.34mmol,3.0 eq) were dissolved in 20mL DMF, then starting material 7 (300 mg,1.34mmol,1.2 eq) was added and reacted for 12 hours at 50deg.C with stirring; TLC (DCM/meoh=20/1, rf/product=0.2) showed that the starting material was reacted completely, a new spot was generated; the reaction was diluted with 100mL ethyl acetate and washed 3 times with saturated brine (100 mL x 3), the organic phase was dried, spin-dried and passed through the column and rinsed with (DCM/meoh=100/1-20/1) to give RDn004 (420 mg, 68.9%) as an off-white solid.
1 H NMR(CDCl3,400MHz)δ:8.16(d,J＝8.7Hz,2H),7.69(d,J＝8.6Hz,1H),7.32(d,J＝8.0Hz,2H),7.24(s,1H),7.20(d,J＝8.8Hz,2H),6.99(dd,J＝8.5,1.8Hz,1H),6.94(d,J＝8.4Hz,2H),4.37(q,J＝8.1Hz,2H),3.87-3.77(m,7H),3.56-3.54(m,2H),2.53-2.50(m,4H).
RDn001, RDn002, RDn003, RDn005, RDn006, RDn007, RDn008, RDn009, RDn010, RDn011, RDn012, RDn013, RDn014, and RDn015 synthetic methods were described in example 1.
RDn001, white solid, yield 75.3%, 1 H NMR(CDCl3,400MHz)δ:8.86(d,J＝8.7Hz,2H),8.53(d,J＝8.6Hz,1H),7.30(d,J＝8.0Hz,2H),7.26(s,1H),7.14(d,J＝8.8Hz,2H),6.80(dd,J＝8.5,1.8Hz,1H),6.90(d,J＝8.4Hz,2H),4.37(q,J＝8.1Hz,2H),3.87-3.77(m,7H),3.56-3.54(m,2H),2.53-2.50(m,4H).
RDn002, pale yellow solid, yield 65.1%, 1 H NMR(CDCl3,400MHz)δ:8.15(d,J＝8.7Hz,2H),7.68(d,J＝8.6Hz,1H),7.30(d,J＝8.0Hz,2H),7.28-7.25(m,4H),7.24(s,1H),7.20(d,J＝8.8Hz,2H),6.92(d,J＝8.4Hz,2H),4.37(q,J＝8.1Hz,2H),3.87-3.77(m,7H),3.56-3.54(m,2H),2.53-2.50(m,4H).
RDn003, white solid, yield 55.7%, 1 H NMR(CDCl3,400MHz)δ:8.88(d,J＝8.7Hz,2H),8.52(d,J＝8.6Hz,1H),7.34(d,J＝8.0Hz,2H),7.24(s,1H),7.12(d,J＝8.8Hz,2H),6.82(dd,J＝8.5,1.8Hz,1H),6.66(d,J＝8.4Hz,2H),4.37(q,J＝8.1Hz,2H),3.87-3.77(m,7H),3.56-3.54(m,2H),2.53-2.50(m,4H).
RDn005, brown solid, yield 32.7%, 1 h NMR (cdcl 3,400 mhz) delta 8.18 (d, j=8.7 hz, 2H), 7.68 (d, j=8.6 hz, 1H), 7.30 (d, j=8.0 hz, 2H), 7.26 (s, 1H), 7.10 (s, 1H), 6.96 (s, 1H), 6.90 (d, j=8.4 hz, 2H), 4.37 (q, j=8.1 hz, 2H), 3.87-3.77 (m, 7H), 3.56-3.54 (m, 2H), 2.53-2.50 (m, 4H) RDn006, white solid, yield 68.9%, 1 H NMR(CDCl3,400MHz)δ:8.88(d,J＝8.7Hz,2H),8.52(d,J＝8.6Hz,1H),7.34-7.24(m,3H),7.12(d,J＝8.8Hz,2H),6.82(dd,J＝8.5,1.8Hz,1H),6.66(d,J＝8.4Hz,2H),4.37(q,J＝8.1Hz,2H),3.87-3.77(m,7H),3.56-3.54(m,2H),2.53-2.50(m,4H).
RDn007, off-white solid, yield 70.2%, 1 H NMR(CDCl3,400MHz)δ:8.86(d,J＝8.7Hz,2H),8.53(d,J＝8.6Hz,1H),7.30(d,J＝8.0Hz,2H),7.26(s,1H),7.14(d,J＝8.8Hz,2H),6.80(dd,J＝8.5,1.8Hz,1H),6.90(d,J＝8.4Hz,2H),4.37(q,J＝8.1Hz,2H),3.87-3.77(m,7H),3.56-3.54(m,2H),2.53-2.50(m,4H).
RDn008, an off-white solid, a yield of 55.6%, 1 H NMR(CDCl3,400MHz)δ:8.15(d,J＝8.7Hz,2H),7.68(d,J＝8.6Hz,1H),7.30(d,J＝8.0Hz,2H),7.28-7.25(m,3H),7.24(s,1H),7.20(d,J＝8.8Hz,2H),6.92(d,J＝8.4Hz,2H),4.37(q,J＝8.1Hz,2H),3.87-3.77(m,7H),3.56-3.54(m,2H),2.53-2.50(m,4H).
RDn009, ashWhite solid, yield 45.8%, 1 H NMR(CDCl3,400MHz)δ:8.17(d,J＝8.7Hz,2H),7.65(d,J＝8.6Hz,1H),7.35(d,J＝8.0Hz,2H),7.30-7.25(m,3H),7.24(s,1H),7.20(d,J＝8.8Hz,2H),6.92(d,J＝8.4Hz,2H),4.37(q,J＝8.1Hz,2H),3.87-3.77(m,7H),3.56-3.54(m,2H),2.53-2.50(m,4H).
RDn010, white solid, yield 45.8%, 1 H NMR(CDCl3,400MHz)δ:8.88(d,J＝8.7Hz,2H),8.52(d,J＝8.6Hz,1H),7.34(d,J＝8.0Hz,2H),7.24(s,1H),7.12(d,J＝8.8Hz,2H),6.82(s,1H),6.66(d,J＝8.4Hz,2H),4.37(q,J＝8.1Hz,2H),4.19(s,3H),3.87-3.77(m,7H),3.56-3.54(m,2H),2.53-2.50(m,4H).
RDn011, white solid, yield 72.8%, 1 H NMR(CDCl3,400MHz)δ:8.88(d,J＝8.7Hz,2H),8.52(d,J＝8.6Hz,1H),7.34(d,J＝8.0Hz,2H),7.24(s,1H),7.12(d,J＝8.8Hz,2H),6.82(s,1H),6.66(d,J＝8.4Hz,2H),4.37(q,J＝8.1Hz,2H),3.87-3.77(m,7H),3.56-3.54(m,2H),2.85(s,6H),2.53-2.50(m,4H).
RDn012, white solid, yield 72.8%, 1 H NMR(CDCl3,400MHz)δ:8.16(d,J＝8.7Hz,2H),7.69(d,J＝8.6Hz,1H),7.32(d,J＝8.0Hz,2H),7.24(s,1H),7.20(d,J＝8.8Hz,2H),6.99(dd,J＝8.5,1.8Hz,1H),6.94(d,J＝8.4Hz,2H),4.37(q,J＝8.1Hz,2H),3.87-3.77(m,7H),3.56-3.54(m,2H),2.53-2.50(m,4H).
RDn013, white solid, yield 58.3%, 1 H NMR(CDCl3,400MHz)δ:8.88(d,J＝8.7Hz,2H),8.52(d,J＝8.6Hz,1H),7.34(d,J＝8.0Hz,2H),7.24(s,1H),7.12(d,J＝8.8Hz,2H),6.82(s,1H),6.66(d,J＝8.4Hz,2H),4.37(q,J＝8.1Hz,2H),4.19(s,3H),3.87-3.77(m,7H),3.56-3.54(m,2H),2.53-2.50(m,4H).
RDn014, light yellow solid, 67.9% yield, 1 H NMR(CDCl3,400MHz)δ:8.88(d,J＝8.7Hz,2H),8.52(d,J＝8.6Hz,1H),7.34(d,J＝8.0Hz,2H),7.24(s,1H),7.12(d,J＝8.8Hz,2H),6.82(dd,J＝8.5,1.8Hz,1H),6.66(d,J＝8.4Hz,2H),4.37(q,J＝8.1Hz,2H),3.87-3.77(m,7H),3.56-3.54(m,2H),2.53-2.50(m,4H).
RDn015, pale yellow solid, yield 67.9%, 1 H NMR(CDCl3,400MHz)δ:8.15(d,J＝8.7Hz,2H),7.68(d,J＝8.6Hz,1H),7.30(d,J＝8.0Hz,2H),7.28-7.25(m,3H),7.24(s,1H),7.20(d,J＝8.8Hz,2H),6.92(d,J＝8.4Hz,2H),4.37(q,J＝8.1Hz,2H),4.18(s,3H),3.87-3.77(m,7H),3.56-3.54(m,2H),2.53-2.50(m,4H).
example 2: proliferation inhibition of cells such as RDn001, RDn002, RDn003, RDn004, RDn005, RDn006, RDn007, RDn008, RDn009, RDn010, RDn011, RDn012, RDn013, RDn014, and RDn015 against gastric cancer, pancreatic cancer, breast cancer, liver cancer, lung cancer, colon cancer, leukemia, lymphoma, and multiple myeloma
Respectively collecting tumor cells in logarithmic growth phase, and adjusting cell suspension concentration to 5×10 4 Each mL was added to a 96-well cell culture plate at a volume of 100uL per well. The novel aryl alkyne compounds RDn001, RDn002, RDn003, RDn004, RDn005, RDn006, RDn007, RDn008, RDn009, RDn010, RDn011, RDn012, RDn013, RDn014 and RDn015 are diluted by DMSO and then added into culture wells, so that the final concentration of the compounds in the system is 0.1, 0.3, 1, 3, 10, 30, 100 and 300 (mu mol/L). After further culturing for 48h, 10. Mu.L of MTT solvent (5 mg/mL) was added to each well, incubated at 37℃for 4h, the culture supernatant was aspirated and discarded, 150. Mu.L of MSO was added to each well, shaking and decolorizing was performed for 10min on a shaker, the reading was performed on the microplate reader, the OD at an absorbance wavelength of 490nm was measured, the results were recorded, and the cell growth curve was drawn with the dose of the compound as the abscissa and the absorbance as the ordinate. The statistical results of half-maximal inhibition (IC 50 value) of tumor cells by RDn001, RDn002, RDn003, RDn004, RDn005, RDn006, RDn007, RDn008, RDn009, RDn010, RDn011, RDn012, RDn013, RDn014, RDn015 and the like are shown in table 1.
Table 1: IC50 values (nM) of RDn series compounds in different tumor cell lines
The results in table 1 show that: RDn001, RDn002, RDn003, RDn004, RDn005, RDn006, RDn007, RDn008, RDn009, RDn010, RDn011, RDn012, RDn013, RDn014 and RDn015 have good proliferation inhibition effect on tumor cells such as gastric cancer, pancreatic cancer, breast cancer, liver cancer, lung cancer, colon cancer, leukemia, lymphoma and multiple myeloma, and particularly, RDn001 has the strongest tumor inhibition activity on lymphoma OCI-LY 3.
Example 3: RDn001 can down regulate STAT3 phosphorylation and target protein expression in OCI-LY3 cells
1. Cell culture and drug addition: taking OCI-LY3 cells in logarithmic growth phase, adjusting to 4×10 density 5 individual/mL of single cell suspension was seeded into 6-well plates at 2mL of cell suspension per well. 37 ℃ and 5% CO 2 Incubators were incubated overnight and RDn001 was added at various concentrations (0, 10, 30, 100 nM). After further incubation for 24h, cells were lysed with RIPA lysate and proteins were collected.
2. Cell collection and lysis: a. the upper medium was discarded and the cells were washed twice with pre-chilled PBS. 100. Mu.L of precooled RIPA cell lysate (protease inhibitor and PMSF were added to the lysate at a ratio of 1:100) was added per well. b. Lysing on ice for 3min, scraping off the cells with a cell scraper and collecting into a 1.5mL EP tube; the mixture was then placed on ice for 30min and vortexed once every 6 min. c.4℃and 12000g for 10min. d. Cell supernatants were transferred to new EP tubes. Cell supernatants are divided into two parts: 5 mu L of the sample is added into an EP tube of 1.5mL for BCA protein content measurement, and 45 mu L of 1 XPBS is added for even mixing; the remaining cell supernatants were quantified by taking 80. Mu.L, adding 5X SDS Loading Buffer. Mu.L, mixing well, boiling in boiling water for 10min, centrifuging, and loading or storing in a refrigerator at-20deg.C.
e. Protein concentration determination: (1) BCA working solution preparation: the total required amount of A and B mixed working fluid is calculated according to the number of the standard substance and the sample to be tested. The volume ratio of the solid agent A to the solid agent B is 50:1, preparing working solution, and uniformly mixing by vortex oscillation for standby.
(2) 1 XPBS diluted protein standard:
(3)
protein standards and sample supernatants diluted with PBS (10-fold dilution) were each taken at 25 μl and added to a new 96-well plate. Then 200. Mu.L of BCA working solution prepared in advance was added respectively and mixed well. The bubbles are not generated by blowing, the cover of the 96-well plate is tightly covered, and the reaction is carried out for 30min in a constant temperature box at 37 ℃. (4) Taking out the 96-well plate, recovering to room temperature for 3-5 min, measuring the absorbance value of A562 on an enzyme label instrument, copying out the obtained value and storing the obtained value in an Excel table. A standard curve was made and the protein content of 1 μl per sample was calculated for protein loading.
3. SDS-PAGE: (1) The gel plate was fixed and 10% SDS-PAGE separating gel was prepared.
The release gel was prepared according to the following table: 10mL
Deionized water 4.0mL
30％(m/v)Acrylamide 3.3mL
1.5M Tris-HCl (pH 8.8) buffer 2.5mL
10％(m/v)SDS 0.1mL
10％(m/v)APS 0.1mL
TEMED 4μL
Total 10mL
(2) And (3) adding the mixed separating glue into 2 glue plates respectively, adding the glue plates to a position 1.0cm away from the top, filling the glue plates with absolute ethyl alcohol, and standing for 30-45 min. (3) After the gel is separated, the residual absolute ethyl alcohol is poured out, and the residual absolute ethyl alcohol is sucked clean by filter paper. (4) 5mL of 5% concentrated gel was prepared according to the following Table
Deionized water 2.77mL
30％(m/v)Acrylamide 830μL
0.5M Tris-HCl (pH 6.8) buffer 1.26mL
10％(m/v)SDS 50μL
10％(m/v)APS 50μL
TEMED 5μL
Total 5mL
(5) Slowly adding the prepared concentrated glue into a glue plate to avoid generating bubbles, inserting a comb, and standing for 30-45 min.
(6) The protein sample was removed and heated in a water bath at 100deg.C for 5min at 10000rpm and centrifuged for 5min. (7) The gel plate was fixed in an electrophoresis tank, SDS-PAGE electrophoresis buffer was added, the comb was pulled out, and the treated protein samples were sequentially added to the sample tank, 50. Mu.g per well of protein. (8) 80V electrophoresis for 40min. (9) Changing the voltage to 120V for electrophoresis for about 1.5 hours until bromophenol blue goes out of the colloid;
4. western-blot: (1) And (3) placing the SDS-PAGE gel subjected to electrophoresis into TBST buffer solution for rinsing once, and placing the protein gel into transfer buffer solution for soaking. (2) Soaking a layer of cotton pad in a membrane transfer buffer solution, clamping onto a membrane transferring instrument by using forceps, placing a blackboard, the cotton pad, filter paper, albumin glue, PVDF membrane, filter paper, the cotton pad and a whiteboard in order, clamping, and placing into the membrane transferring instrument. If bubbles exist between each two layers, the bubbles are removed by using a glass solid tube to lightly roll. And (3) opening a film transfer instrument, and performing constant-current transfer printing for 80 minutes at 300 mA. (4) The membranes were placed in TBST buffer and rinsed 3 times for 8min each. (5) blocking with 20mL of 5% BSA-TBST blocking solution at room temperature for 2h. (6) Primary antibody was added and incubated at4℃at 60rpm overnight. (7) The membranes were washed three times with TBST at room temperature, 60rpm, 10min each. (8) adding a secondary antibody, and incubating for 1h at room temperature. (9) The membranes were washed three times with TBST at room temperature, 60rpm, 10min each. (10) 1mL of each of chemiluminescent substrate solid solution A and chemiluminescent substrate solid solution B is taken and developed for 2min at room temperature. (11) The liquid on the membrane was blotted with filter paper and exposed on the machine, the results are detailed in FIG. 1. 5. And (3) preparation of a reagent:
(1) 10% SDS: 1g of high purity (electrophoretic grade) SDS was weighed into a 10mL centrifuge tube, added with about 8mL deionized water, dissolved by heating, and stored at room temperature to a volume of 10 mL.
(2) 10% ammonium persulfate (Ammonium persulfate, AP): 1g of ammonium persulfate was weighed, dissolved by stirring after adding about 10mL of deionized water, and stored at4 ℃.
(3) 5 Xrunning buffer: 15.1g of Tris, 94g of Glycine and 5.0g of SDS are weighed in a beaker, added with 1L of double distilled water for dissolution, stored at room temperature and diluted 5 times with time.
(4) Transfer buffer: 5.8g of Tris, 11.6g of glycine and 0.75g of SDS are weighed into a beaker, 700mL of double distilled water is added, the volume is fixed to 800mL after dissolution, and finally 200mL of methanol is added.
(5) 1.5mol/L Tris-HCl,100mL: then 18.15gtris is dissolved in 80mL water and adjusted to 8.8 with 4N HCl to a volume of 100 mL.
(6) 0.5mol/L Tris-HCl,1000mL: 60.5 g of tris base was weighed, water was added to 850ml, concentrated hydrochloric acid was added and stirred until all dissolved, the pH was adjusted to 6.8, and water was added to 1L.
(7) TBS buffer: 8.8g of NaCl was weighed out in 800mL of distilled water, dissolved, 10mL of 1mol/L TrisHCl (pH 7.5) was added, the volume was fixed to 1L, and the mixture was stored at room temperature.
(8) TBST buffer: to 1L TBS buffer, 500. Mu.L of 20% Tween20 was added to give a final concentration of 0.1% Tween20, and the mixture was prepared immediately.
(9) Blocking solution, antibody dilution: 5% skim milk powder or BSA was added to TBST buffer and ready-to-use.
The results in FIG. 1 show that RDn001 treatment at 10nM, 30nM and 100nM is effective in down-regulating the expression levels of p-STAT3 (Y705), p-STAT3 (S727) and the target proteins Cyclin D1 and BCL-XL.
Taken together, the results show that the aryl alkyne compound represented by RDn001 can obviously inhibit the growth of gastric cancer, pancreatic cancer, breast cancer, liver cancer, lung cancer, colon cancer, leukemia, lymphoma and multiple myeloma cells, has obvious inhibition effect on STAT3 and related proteins thereof, and shows good anticancer effect. According to the general way of drug development (conventional anti-tumor in-vitro screening is carried out firstly and then targeted research is carried out), the compound can be applied to cancer treatment drugs related to abnormal cell proliferation, and the anti-tumor drugs can be prepared by salifying with human bodies or mixing with a medicinal carrier.
Finally, what should be said is: the above embodiments are only for illustrating the technical solution of the present invention, but any equivalent replacement of the present invention and modification or partial replacement without departing from the spirit and scope of the present invention should be covered in the scope of the claims of the present invention.

Claims (7)

1. An aryl alkyne compound is characterized in that the structural formula is shown in a general formula I:
wherein the A ring is selected from
The B ring is selected from
2. The aryl alkyne compound according to claim 1, wherein the compound has the following structure:
3. the biologically acceptable salt of an arylalkyne compound of claim 1 or 2 with at least one of acetic acid, dihydrofolate, benzoic acid, citric acid, sorbic acid, propionic acid, oxalic acid, fumaric acid, maleic acid, hydrochloric acid, malic acid, phosphoric acid, sulfurous acid, sulfuric acid, vanilloid acid, tartaric acid, ascorbic acid, boric acid, lactic acid, and ethylenediamine tetraacetic acid.
4. The process for the preparation of aryl acetylenes according to claim 1, characterized in that the synthetic route is as follows:
the synthesis steps are as follows
(1) Dissolving the compound 1, the compound 2, DIEA and HBTU in DMF, stirring at room temperature to react completely, pouring the reaction solution into water, precipitating, filtering, collecting solid and drying to obtain a compound 3;
(2) Compound 3, compound 4, triethylamine and Pd (PPh 3 ) 2 Cl 2 Dissolving CuI in DMF, stirring at 70-90 ℃ for complete reaction, diluting the reaction solution with ethyl acetate, washing with saline solution, drying the organic phase, spin-drying, stirring, and passing through a column to obtain a compound 5;
(3) Dissolving the compound 5 in methanol, adding an ethyl acetate solution of hydrogen chloride, stirring at room temperature to react completely, and directly spin-drying the reaction solution to obtain a compound 6;
(4) Dissolving the compound 6 and potassium carbonate in DMF, adding the compound 7, stirring at 40-60 ℃ for complete reaction, diluting the reaction liquid with ethyl acetate, washing with saline solution, drying the organic phase, spin-drying, stirring, and passing through a column to obtain the compound shown in the formula I.
5. The process for producing an arylalkyne compound according to claim 4, wherein in step (1), the molar ratio of compound 1 to compound 2 to DIEA to HBTU is 1:1:2, (1.0 to 1.5); in step (2), compound 3, compound 4, triethylamine and Pd (PPh 3 ) 2 Cl 2 And CuI in a molar ratio of 1:2:3:0.02:0.01; in the step (3), the molar ratio of the compound 5 to the hydrogen chloride is 1:10; in the step (4), the molar ratio of the compound 6 to the potassium carbonate to the compound 7 is 1 (2-4): 1.0-1.5.
6. Use of an arylalkyne compound according to any one of claims 1 to 3, and of its biologically acceptable salts, for the manufacture of an antitumor medicament for the treatment of diseases associated with abnormal STAT3 cell signalling.
7. The use according to claim 6, wherein the medicament for treating diseases associated with abnormal STAT3 cell signaling is a medicament for treating gastric cancer, pancreatic cancer, breast cancer, liver cancer, lung cancer, colon cancer, leukemia, lymphoma and multiple myeloma.
CN202311519441.8A 2023-11-15 2023-11-15 Aryl alkyne compound as well as preparation method and application thereof Pending CN117567361A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202311519441.8A CN117567361A (en) 2023-11-15 2023-11-15 Aryl alkyne compound as well as preparation method and application thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202311519441.8A CN117567361A (en) 2023-11-15 2023-11-15 Aryl alkyne compound as well as preparation method and application thereof

Publications (1)

Publication Number Publication Date
CN117567361A true CN117567361A (en) 2024-02-20

Family

ID=89892858

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202311519441.8A Pending CN117567361A (en) 2023-11-15 2023-11-15 Aryl alkyne compound as well as preparation method and application thereof

Country Status (1)

Country Link
CN (1) CN117567361A (en)

Similar Documents

Publication Publication Date Title
US20220098162A1 (en) Aphthylamine compound and biologically acceptable salt thereof, preparation method therefor, and application thereof
CN113603707B (en) Tricyclic heteroaryl-containing compounds
CN114702439B (en) Naphthyl urea-piperazine compound and preparation method and application thereof
EP4011867B1 (en) Crystal forms c and e of pyrazin-2(1h)-one compound and preparation method therefor
KR102507328B1 (en) Crystal forms and salt forms of TGF-βRI inhibitors and methods for their preparation
CN114269723B (en) D crystal form of pyrazine-2 (1H) -ketone compound and preparation method thereof
CN111253339B (en) Synthetic preparation method of curcumin derivatives and application of curcumin derivatives in cancer treatment
EP4011866A1 (en) A crystalline form and b crystalline form of pyrazine-2(1h)-ketone compound and preparation method thereof
WO2020253875A1 (en) Salts of compound, crystalline forms thereof, preparation method therefor and application thereof
CN109369620B (en) Pyridine compound, preparation method thereof and application thereof in resisting gastric cancer
CN110437156B (en) Paeonol dihydropyrimidinone derivative, preparation method and application thereof
CN117567361A (en) Aryl alkyne compound as well as preparation method and application thereof
CN114292270B (en) BTK inhibitor and preparation method and application thereof
JP7101781B2 (en) Salt morphology as an Akt inhibitor and its crystalline morphology
CN108530337B (en) Indoleamide compound capable of selectively inhibiting gastric cancer cells
CN108047182B (en) Daphnoretin derivative and application thereof
CN108623511A (en) A kind of indole amides class compound can be used for treating cancer
CN110078706B (en) Imatinib derivative and preparation method and application thereof
CN117263851A (en) Pyridine-naphthyl urea-piperazine derivative, and preparation method and application thereof
CN116178248A (en) Naphthyl urea-piperidine compounds, and preparation method and application thereof
JP2022533440A (en) Crystal forms of c-MET/AXL inhibitors
CN118063410A (en) Piperazine naphthalimide compound, and preparation method and application thereof
CN115304605B (en) Oxetane derivatives with antitumor activity, and preparation method and application thereof
CN114105892B (en) FAK/PLK1 double-target quinazoline derivative and preparation method and application thereof
CN114853751B (en) Group of phenothiazine derivatives and application thereof

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination