CN118126038B - Pyrazolopyridine derivative, and preparation method and application thereof - Google Patents

Pyrazolopyridine derivative, and preparation method and application thereof Download PDF

Info

Publication number
CN118126038B
CN118126038B CN202410559979.XA CN202410559979A CN118126038B CN 118126038 B CN118126038 B CN 118126038B CN 202410559979 A CN202410559979 A CN 202410559979A CN 118126038 B CN118126038 B CN 118126038B
Authority
CN
China
Prior art keywords
compound
pyrazolopyridine derivative
reaction
pyrazolopyridine
derivative according
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.)
Active
Application number
CN202410559979.XA
Other languages
Chinese (zh)
Other versions
CN118126038A (en
Inventor
殷齐坤
刘义
谭深鹏
王洪波
卢嘉国
宣轩
董智文
李蕙荣
叶泰强
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.)
Yantai University
Original Assignee
Yantai University
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 Yantai University filed Critical Yantai University
Priority to CN202410559979.XA priority Critical patent/CN118126038B/en
Publication of CN118126038A publication Critical patent/CN118126038A/en
Application granted granted Critical
Publication of CN118126038B publication Critical patent/CN118126038B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
    • C07D471/04—Ortho-condensed systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • General Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Nitrogen Condensed Heterocyclic Rings (AREA)

Abstract

The invention discloses pyrazolopyridine derivatives, a preparation method and application thereof, and belongs to the technical field of pharmaceutical chemistry. The pyrazolopyridine derivative has the following structure: Or (b) . The derivatives have good inhibition effects on human pancreatic cancer cells PANC-1, gastric cancer cells AGS, triple negative breast cancer cells HCC1806 and HCC1937, and the IC 50 value of the optimal molecule is in the range of 0.6-2.5 mu M, so that the derivatives can be used as antitumor lead molecules.

Description

Pyrazolopyridine derivative, and preparation method and application thereof
Technical Field
The invention relates to a compound and a preparation method and application thereof, in particular to pyrazolopyridine derivative and a preparation method thereof and application thereof in preparing medicines for treating pancreatic cancer, gastric cancer or breast cancer, and belongs to the technical field of pharmaceutical chemistry.
Background
Cancer is a serious disease threatening human health, is also a global serious public health problem at present, and according to World Health Organization (WHO) 2023 data statistics, the number of newly increased cancer cases in the world exceeds 2000 tens of thousands, the number of death cases is nearly 1000 tens of thousands, and the global cancer burden is increasingly increased. Based on various factors such as indication, treatment cost, safety and the like, chemotherapy is the primary strategy of the current tumor treatment, and the development of chemical small molecules with anti-tumor activity is the main strategy of the discovery of the current novel anti-tumor drugs. The nitrogen-containing heterocycle and the aromatic skeleton are one of the important structures constituting the medicine, and the chemical molecule ratio of the nitrogen-containing heterocycle and the aromatic skeleton is 88% and 87% respectively in 164 chemical small molecules approved by the U.S. Food and Drug Administration (FDA) in the last five years. Pyrazolopyridine is a common and important nitrogenous heterocyclic derivative, contains steric hindrance and electronic arrangement similar to indole, azaindole and other structures, and has various pharmacological activities such as anti-inflammatory, antibacterial, antitumor and the like. The invention is based on pyrazolopyridine parent nucleus to develop the preparation of molecules with anti-tumor activity and the application evaluation of the anti-tumor activity.
Disclosure of Invention
The invention aims at: pyrazolopyridine derivatives, a preparation method thereof and application thereof in preparing medicaments for treating pancreatic cancer, gastric cancer or breast cancer are provided.
In order to achieve the above object, the present invention adopts the following technical scheme:
pyrazolopyridine derivative has the following structure:
Or (b) ；
Wherein, R 1 is selected from any one of the following structures:
；
r 2 is selected from any one of the following structures:
。
the preparation method of the pyrazolopyridine derivative comprises the following steps:
Step 1, cyclizing: dissolving 5-bromo-2-chloronicotinonitrile in absolute ethyl alcohol, heating to 85 ℃, dropwise adding hydrazine hydrate, maintaining the temperature of a reaction system, stirring, and after the reaction is finished, sequentially carrying out low-temperature recrystallization, vacuum filtration and drying to obtain a compound 2; preferably, the dosage ratio of the 5-bromo-2-chloronicotinonitrile to the hydrazine hydrate is 23mmol:115mmol;
Step 2, carbon-carbon coupling: dissolving a compound 2, a compound A and potassium phosphate in dioxane aqueous solution, wherein the compound A is a compound 3, a compound 9, a compound 12 or a compound 15, blowing nitrogen, adding ditri-butyl phosphine palladium into a reaction system, reacting at 80-110 ℃, and sequentially carrying out diatomite vacuum filtration, ethyl acetate and water extraction and drying after the reaction is finished to obtain a compound B;
Step 3, amidation: dissolving a compound B in N, N-dimethylformamide, adding a compound C, wherein the compound C is bromoacetyl chloride, chloroacetyl chloride, 2-trichloroethyl chloroformate or isobutyryl chloride, reacting at 20 ℃, and sequentially quenching, extracting with ethyl acetate and water, performing vacuum rotary evaporation, passing through a column and drying after the reaction is finished to obtain the pyrazolopyridine derivative of claim 1;
Wherein the structures of compound 2, compound 3, compound 9, compound 12 and compound 15 are shown below, respectively:
、、、、。
preferably, in step 2, the ratio of the amounts of compound 2, compound a, potassium phosphate and ditri-butyl phosphine palladium is 14mmol:16.8mmol:28mmol:0.7mmol.
Preferably, in step 3, the ratio of compound B to compound C is 0.3mmol:0.6mmol.
More preferably, the method further comprises a methylation step, in particular:
After the step 2 is finished, the compound B is dissolved in N, N-dimethylformamide, sodium hydride is added under ice bath, then potassium iodide is added dropwise for reaction, quenching, ethyl acetate and water extraction, vacuum rotary evaporation, column passing and drying are sequentially carried out after the reaction is finished, the compound D is obtained, methylation is finished, and then the compound D is substituted for the compound B for amidation reaction.
Preferably, in the methylation step, the ratio of the amounts of compound B, sodium hydride and potassium iodide is 0.3mmol:0.45mmol:0.45mmol.
The pyrazolopyridine derivative is applied to the preparation of anticancer drugs, wherein the anticancer drugs are pancreatic cancer resistant drugs, gastric cancer resistant drugs or breast cancer resistant drugs.
The invention has the advantages that:
The pyrazolopyridine derivatives with novel structures are developed, the derivatives have good inhibition effects on human pancreatic cancer cells PANC-1, gastric cancer cells AGS and triple negative breast cancer cells HCC1806 and HCC1937, the IC 50 value of the optimal molecules is in the range of 0.6-2.5 mu M, and the optimal molecules can be used as antitumor lead molecules;
(2) The pyrazolopyridine derivative with the novel structure can be obtained by taking a commercial product (5-bromo-2-chloronicotinonitrile) as a substrate through three-step chemical reaction (cyclization, carbon-carbon coupling and amidation) or four-step chemical reaction (cyclization, carbon-carbon coupling, methylation and amidation), and the preparation method has the advantages of low cost, simple condition, easiness in separation and the like.
Drawings
FIGS. 1 and 2 are respectively the hydrogen and carbon spectra of Compound 5;
FIGS. 3 and 4 are respectively the hydrogen and carbon spectra of Compound 6;
FIGS. 5 and 6 are respectively the hydrogen and carbon spectra of Compound 7;
FIGS. 7 and 8 are respectively the hydrogen and carbon spectra of Compound 8;
FIGS. 9 and 10 are respectively the hydrogen and carbon spectra of Compound 11;
FIGS. 11 and 12 are respectively the hydrogen and carbon spectra of Compound 14;
FIGS. 13 and 14 are respectively the hydrogen and carbon spectra of Compound 17;
FIGS. 15 and 16 are respectively the hydrogen and carbon spectra of Compound 19;
Fig. 17 is a graph showing the effect of compound 6 on apoptosis of human triple negative breast cancer cells HCC1937 at various concentrations.
Detailed Description
The invention is described in detail below with reference to the drawings and the specific embodiments.
1. Structure of pyrazolopyridine derivative
The pyrazolopyridine derivative provided by the invention has the following structure:
Or (b) ；
Wherein, R 1 is selected from any one of the following structures:
；
r 2 is selected from any one of the following structures:
。
2. pyrazolopyridine derivative preparation method
Example 1
23Mmol of 5-bromo-2-chloronicotinonitrile (compound 1) was dissolved in 150mL of absolute ethanol (EtOH), the reaction system was heated to 85℃and 115mmol of hydrazine hydrate (H 2N-NH2) was added dropwise, the temperature of the reaction system was maintained and the reaction was stirred for 2H. After the reaction, the reaction system was recrystallized at low temperature in an ice-water bath, filtered under vacuum, and dried to give compound 2 (yellow crystals) in 83% crude yield.
14Mmol of compound 2, 16.8mmol of compound 3 and 28mmol of potassium phosphate were dissolved in 80mL of dioxane aqueous solution (dioxane: water=2:1, v/v), nitrogen was bubbled for 10min, 0.7mmol of di-tert-butylphosphine palladium was added to the reaction system, and the mixture was reacted at 110℃for 72h. After the reaction, the reaction system was vacuum filtered with celite, extracted with ethyl acetate and water, and dried to give compound 4 in 68% crude yield.
0.3Mmol of compound 4 was dissolved in 3mL of N, N-Dimethylformamide (DMF), and 0.6mmol of bromoacetyl chloride was added thereto for reaction at 20℃for 3 hours. After the reaction, the reaction system was quenched with water, extracted with ethyl acetate and water, and subjected to rotary evaporation under vacuum and column chromatography with V Dichloromethane (dichloromethane) /V Methanol =50:1, followed by drying to give compound 5 (white solid) in 47% yield.
The hydrogen spectrum and the carbon spectrum of the compound 5 are shown in fig. 1 and 2 respectively, and the information of each spectrum is specifically as follows:
1H NMR（d6-DMSO,400MHz）δ13.54（s,1H）,11.19（s,1H）,8.92（d,J=2Hz,1H）,8.72（d,J=2.Hz,1H）,8.02-7.95（m,4H）,4.41（s,2H）,3.49-3.42（m,1H）,1.20（s,3H）,1.18（s,3H）;
13C NMR（d6-DMSO,100MHz）δ165.5,152.2,149.4,143.7,140.2,136.0,131.7,130.0（2C）,128.2（2C）,127.5,108.1,54.8,43.4,15.8（2C）.
The calculated hydrogenation value of the molecular weight of the compound 5 (2-bromo-N- (5- (4- (isopropylsulfonyl) phenyl) -1H-pyrazolo [3,4-b ] pyridin-3-yl) acetamide) is 437.1929 and the theoretical hydrogenation value is 437.0283 through detection of high resolution mass spectrum (HR-MS).
Example 2
Compound 4 was prepared by the same method as in example 1.
0.3Mmol of compound 4 was dissolved in 3mL of N, N-Dimethylformamide (DMF), and 0.6mmol of chloroacetyl chloride was added thereto for reaction at 20℃for 3 hours. After the reaction was completed, the reaction system was quenched with water, extracted with ethyl acetate and water, and subjected to rotary evaporation under vacuum and column chromatography with V Dichloromethane (dichloromethane) /V Methanol =50:1, followed by drying to give compound 6 (white solid) in 48% yield.
The hydrogen spectrum and the carbon spectrum of the compound 6 are respectively shown in fig. 3 and 4, and the information of each spectrum is specifically as follows:
1H NMR（d6-DMSO,400MHz）δ 13.53（s,1H）,11.18（s,1H）,8.91（d,J=2Hz,1H）,8.72（d,J=2Hz,1H）,8.01-7.95（m,4H）,4.41（s,2H）,3.49-3.42（m,1H）,1.20（s,3H）,1.18（s,3H）;
13C NMR（d6-DMSO,100MHz）δ165.5,152.2,149.4,143.7,140.2,136.0,131.7,130.0（2C）,128.2,127.5,108.1,54.8,15.8 2.
the molecular weight of the compound 6 (2-chloro-N- (5- (4- (isopropylsulfonyl) phenyl) -1H-pyrazolo [3,4-b ] pyridin-3-yl) acetamide) is detected by high resolution mass spectrometry (HR-MS) and has a hydrogenation calculated value of 393.0775 and a hydrogenation theoretical value of 393.0783.
Example 3
Compound 4 was prepared by the same method as in example 1.
0.3Mmol of compound 4 was dissolved in 3mL of N, N-Dimethylformamide (DMF), and 0.6mmol of 2, 2-trichloroethyl chloroformate was added to react at 20℃for 3 hours. After the reaction was completed, the reaction system was quenched with water, extracted with ethyl acetate and water, and subjected to rotary evaporation under vacuum and column chromatography with V Dichloromethane (dichloromethane) /V Methanol =50:1, followed by drying to give compound 7 (white solid) in 49% yield.
The hydrogen spectrum and the carbon spectrum of the compound 7 are shown in fig. 5 and 6 respectively, and the information of each spectrum is specifically as follows:
1H NMR（d6-DMSO,400MHz）δ13.47（s,1H）,10.84（s,1H）,8.92（d,J=2Hz,1H）,8.64（d,J=2Hz,1H）,8.03-7.95（m,4H）,5.00（s,2H）,3.50-3.43（m,1H）,1.20（s,3H）,1.19（s,3H）;
13C NMR（d6-DMSO,100MHz）δ153.2,152.1,149.4,143.7,139.7,136.1,130.0（2C）,128.3（2C）,127.5,108.4,96.3,74.4,54.7,15.8（2C）.
The calculated hydrogenation value of the molecular weight of the compound 7 (2, 2-trichloroethyl (5- (4- (isopropyl sulfonyl) phenyl) -1H-pyrazolo [3,4-b ] pyridin-3-yl) carbamate) is 491.0108 and the theoretical hydrogenation value is 491.0109 through detection of high-resolution mass spectrum (HR-MS).
Example 4
Compound 4 was prepared by the same method as in example 1.
0.3Mmol of Compound 4 was dissolved in 3mL of N, N-Dimethylformamide (DMF), and 0.6mmol of isobutyryl chloride was added thereto to react at 20℃for 3 hours. After the reaction was completed, the reaction system was quenched with water, extracted with ethyl acetate and water, and subjected to rotary evaporation under vacuum and column chromatography with V Dichloromethane (dichloromethane) /V Methanol =50:1, followed by drying to give compound 8 (white solid) in 37% yield.
The hydrogen spectrum and the carbon spectrum of the compound 8 are shown in fig. 7 and 8 respectively, and the information of each spectrum is specifically as follows:
1H NMR（d6-DMSO,400MHz）δ13.38（s,1H）,10.72（s,1H）,8.88（d,J=2Hz,1H）,8.73（d,J=2Hz,1H）,8.00-7.95（m,4H）,3.49-3.42（m,1H）,2.81-2.74（m,1H）,1.20（s,3H）,1.19（s,3H）,1.17（s,3H）,1.15（s,3H）;
13C NMR（d6-DMSO,100MHz）δ175.9,152.2,149.2,143.9,141.1,135.9,132.2,130.0（2C）,128.3（2C）,127.2,108.2,54.8,34.5,20.0（2C）,15.8（2C）.
the calculated sodium addition for the molecular weight of compound 8 (N- (5- (4- (isopropylsulfonyl) phenyl) -1H-pyrazolo [3,4-b ] pyridin-3-yl) isobutyramide) was 409.0903 and the theoretical sodium addition was 409.1305 as determined by high resolution mass spectrometry (HR-MS).
Example 5
Compound 2 was prepared by the same method as in example 1.
14Mmol of compound 2, 16.8mmol of compound 9 and 28mmol of potassium phosphate were dissolved in 80mL of dioxane aqueous solution (dioxane: water=2:1, v/v), nitrogen was bubbled for 10min, 0.7mmol of di-tert-butylphosphine palladium was added to the reaction system, and the mixture was reacted at 80℃for 72h. After the reaction, the reaction system was vacuum filtered with celite, extracted with ethyl acetate and water, and dried to give compound 10 in 62% crude yield.
0.3Mmol of compound 10 was dissolved in 3mL of N, N-Dimethylformamide (DMF), and 0.6mmol of chloroacetyl chloride was added thereto for reaction at 20℃for 3 hours. After the reaction was completed, the reaction system was quenched with water, extracted with ethyl acetate and water, and subjected to rotary evaporation under vacuum and column chromatography with V Dichloromethane (dichloromethane) /V Methanol =50:1, followed by drying to give compound 11 (white solid) in 62% yield.
The hydrogen spectrum and the carbon spectrum of the compound 11 are shown in fig. 9 and 10 respectively, and the information of each spectrum is specifically as follows:
1H NMR（d6-DMSO,400MHz）δ13.37（s,1H）,11.09（s,1H）,8.78（d,J=2Hz,1H）,8.50（d,J=2Hz,1H）,7.62（d,J=8.8Hz,2H）,7.07（d,J=8.8Hz,2H）,4.40（s,2H）,3.80（s,3H）;
13C NMR（d6-DMSO,100MHz）δ165.4,159.5,151.7,149.1,139.6,130.8,129.7,129.1,128.7（2C）,115.2（2C）,108.2,55.7,43.4.
The calculated value of the molecular weight of the compound 11 (2-chloro-N- (5- (4-methoxyphenyl) -1H-pyrazolo [3,4-b ] pyridin-3-yl) acetamide) is 339.0617 and the theoretical value of sodium is 339.0619 through detection of high-resolution mass spectrum (HR-MS).
Example 6
Compound 2 was prepared by the same method as in example 1.
14Mmol of compound 2, 16.8mmol of compound 12 and 28mmol of potassium phosphate were dissolved in 80mL of dioxane aqueous solution (dioxane: water=2:1, v/v), nitrogen was bubbled for 10min, 0.7mmol of di-tert-butylphosphine palladium was added to the reaction system, and the mixture was reacted at 80℃for 72h. After the reaction was completed, the reaction system was vacuum filtered with celite, extracted with ethyl acetate and water, and dried to give compound 13 in 83% crude yield.
0.3Mmol of compound 13 was dissolved in 3mL of N, N-Dimethylformamide (DMF), and 0.6mmol of chloroacetyl chloride was added thereto for reaction at 20℃for 3 hours. After the reaction was completed, the reaction system was quenched with water, extracted with ethyl acetate and water, and subjected to rotary evaporation under vacuum and column chromatography with V Dichloromethane (dichloromethane) /V Methanol =50:1, followed by drying to give compound 14 (white solid) in 54% yield.
The hydrogen spectrum and the carbon spectrum of the compound 14 are shown in fig. 11 and fig. 12 respectively, and the information of each spectrum is specifically as follows:
1H NMR（d6-DMSO,400MHz）δ13.37（s,1H）,11.09（s,1H）,8.78（d,J=2Hz,1H）,8.50（d,J=2Hz,1H）,7.59（d,J=8.4Hz,2H）,7.04（d,J=8.4Hz,2H）,4.68-4.62（m,1H）,4.40（s,2H）,1.29（s,3H）,1.28（s,3H）;
13C NMR（d6-DMSO,100MHz）δ165.4,157.7,151.6,149.1,139.6,130.5,129.7,129.1,128.7（2C）,116.8（2C）,108.2,69.8,43.4,22.4（2C）.
The calculated hydrogenation value of the molecular weight of the compound 14 (2-chloro-N- (5- (4-isopropoxyphenyl) -1H-pyrazolo [3,4-b ] pyridin-3-yl) acetamide) is 34.1104 and the theoretical hydrogenation value is 345.113 through detection of high resolution mass spectrum (HR-MS).
Example 7
Compound 2 was prepared by the same method as in example 1.
14Mmol of compound 2, 16.8mmol of compound 15 and 28mmol of potassium phosphate were dissolved in 80mL of dioxane aqueous solution (dioxane: water=2:1, v/v), nitrogen was bubbled for 10min, 0.7mmol of di-tert-butylphosphine palladium was added to the reaction system, and the mixture was reacted at 80℃for 72h. After the reaction, the reaction system was vacuum filtered with celite, extracted with ethyl acetate and water, and dried to give compound 16 in 85% crude yield.
0.3Mmol of compound 16 was dissolved in 3mL of N, N-Dimethylformamide (DMF), and 0.6mmol of chloroacetyl chloride was added thereto for reaction at 20℃for 3 hours. After the reaction was completed, the reaction system was quenched with water, extracted with ethyl acetate and water, and subjected to rotary evaporation under vacuum and column chromatography with V Dichloromethane (dichloromethane) /V Methanol =50:1, followed by drying to give compound 17 (white solid) in 50% yield.
The hydrogen spectrum and the carbon spectrum of the compound 17 are shown in fig. 13 and 14 respectively, and the information of each spectrum is specifically as follows:
1H NMR（d6-DMSO,400MHz）δ13.50（s,1H）,11.16（s,1H）,8.89（d,J=2Hz,1H）,8.68（d,J=2Hz,1H）,7.96-7.89（m,4H）,7.60（s,1H）,4.41（s,2H）,3.33（s,1H）,1.13（s,9H）;
13C NMR（d6-DMSO,100MHz）δ165.5,152.1,149.4,143.7,141.8,140.1,131.3,127.9（2C）,127.8,127.7（2C）,108.1,53.9,43.4,30.3（3C）.
The calculated hydrogenation for the molecular weight of compound 17 (N- (5- (4- (N- (tert-butyl) sulfamoyl) cyclohex-2, 4-dien-1-yl) -1H-pyrazolo [3,4-b ] pyridin-3-yl) -2-chloroacetamide) was 424.1011, the theoretical hydrogenation value was 424.1205, as determined by high resolution mass spectrometry (HR-MS).
Example 8
Compound 4 was prepared by the same method as in example 1.
0.3Mmol of Compound 4 was dissolved in 3mL of N, N-Dimethylformamide (DMF), and 0.45mmol of sodium hydride was added under ice bath, followed by dropwise addition of 0.45mmol of potassium iodide for 2 hours. After the reaction was completed, the reaction system was quenched with water, extracted with ethyl acetate and water, and subjected to rotary evaporation under vacuum and column chromatography with V Dichloromethane (dichloromethane) /V Methanol =50:1, followed by drying to give compound 18 (yellow solid) in 48% yield.
0.3Mmol of compound 18 was dissolved in 3mL of N, N-Dimethylformamide (DMF), and 0.6mmol of chloroacetyl chloride was added thereto for reaction at 20℃for 3 hours. After the reaction was completed, the reaction system was quenched with water, extracted with ethyl acetate and water, and subjected to rotary evaporation under vacuum and column chromatography with V Dichloromethane (dichloromethane) /V Methanol =50:1, followed by drying to give compound 19 (white solid) in 42% yield.
The hydrogen spectrum and the carbon spectrum of the compound 19 are shown in fig. 15 and fig. 16 respectively, and the information of each spectrum is specifically as follows:
1H NMR（d6-DMSO,400MHz）δ11.24（s,1H）,8.95（d,J=2Hz,1H）,8.73（d,J=2Hz,1H）,8.01-7.96（m,4H）,4.41（s,2H）,3.01（s,3H）,3.50-3.43（m,1H）,1.20（s,3H）,1.19（s,3H）;
13C NMR（d6-DMSO,100MHz）δ165.4,150.3,149.4,143.6,138.9,136.1,132.1,130.0（2C）,128.3（2C）,127.4,108.4,54.8,43.4,34.0,15.8（2C）.
The calculated hydrogenation value of the molecular weight of the compound 19 (2-chloro-N- (5- (4- (isopropylsulfonyl) phenyl) -1-methyl-1H-pyrazolo [3,4-b ] pyridin-3-yl) acetamide) is 407.0941 and the theoretical hydrogenation value is 407.0939 through detection of high resolution mass spectrum (HR-MS).
3. Application evaluation of anticancer Activity of pyrazolopyridine derivative
1. Cell proliferation inhibition assay
The pyrazolopyridine derivatives (compound 5, compound 6, compound 7, compound 8, compound 11, compound 14, compound 17, compound 19) were tested for their ability to inhibit cell proliferation in vitro using a thiazolyl blue (MTT) colorimetric method on human pancreatic cancer cells PANC-1, human gastric cancer cell lines AGS, triple negative breast cancer cells HCC1806 and HCC 1937.
Experimental principle: MTT can be reduced into insoluble blue-violet formazan crystals by amber dehydrogenase in mitochondria of living cells, and the crystals can be dissolved by dimethyl sulfoxide (DMSO) and detected at 570nm wavelength by an enzyme-labeled instrument, and the absorbance reflects the cell survival rate.
Half maximal inhibitory concentration (IC 50) refers to: the concentration of drug required for 50% cell death during a period of tumor cell exposure to drug allows for the calculation of IC 50 values by fitting a dose response curve by measuring the MTT absorbance at different concentrations of drug administered.
The measurement method is as follows:
(1) Tumor cells were seeded at 4000 cells/well in 96 well plates and incubated in a CO 2 incubator for 24h;
(2) The compound to be tested is diluted by 25 mu M of the half-dilution and incubated with tumor cells for 48 hours, and three groups of parallel holes are arranged at each concentration;
(3) After drug treatment, 20. Mu.L MTT solution (2.5 mg/mL) was added to each well plate and incubated for 2h at 37℃in an incubator;
(4) Removing supernatant, adding 150 mu L DMSO into each well, and shaking thoroughly until formazan is completely dissolved;
(5) And detecting the absorbance of the sample at 570nm by using an enzyme-labeled instrument, fitting a dose response curve, and calculating the IC 50 value of the compound to be detected.
The results of toxicity tests on human pancreatic cancer cells PANC-1, human gastric cancer cell lines AGS, triple negative breast cancer cells HCC1806 and HCC1937 after 48 hours of administration of the above compounds are shown in Table 1.
Toxicity test results (IC 50 value, μM) of each compound of Table 1 on three tumor cells
Cell proliferation inhibition experiments show that the compounds developed by the invention have good in vitro tumor inhibition effect on pancreatic cancer, gastric cancer and triple negative breast cancer. Wherein:
(1) Compound 6 has the best inhibitory effect on triple negative breast cancer cell HCC1806 with IC 50 value of 2.4 μm;
(2) Compound 6, compound 11, compound 14, compound 17 and compound 19 have the best inhibitory effect on triple negative breast cancer cell HCC1937, with IC 50 values of 0.6 μm, 1.9 μm, 1.1 μm, 0.7 μm, 1.3 μm, respectively;
(3) Compound 5, compound 6, compound 7, compound 17 and compound 19 have the best inhibitory effect on human pancreatic cancer cells PANC-1, with IC 50 values of 2.5 μm, 2.6 μm, 1.3 μm, 0.7 μm, respectively;
(4) Compound 6 and compound 14 have the best inhibitory effect on human gastric cancer cell line AGS, with IC 50 values of 1.2 μm and 1.1 μm, respectively;
(5) The compound 6 has better inhibition effect on triple negative breast cancer cells HCC1806 and HCC1937, human pancreatic cancer cells PANC-1 and human gastric cancer cell strain AGS, and is a preferable compound.
2. Apoptosis experiments
The effect of the preferred compound (compound 6) on apoptosis in triple negative breast cancer cells HCC1937 was examined in vitro using flow cytometry.
Experimental principle: phosphatidylserine (PS) of normal cells is located inside the cell membrane, and when early apoptosis occurs, PS inside the membrane will evert to the surface of the cell membrane and bind to fluorescein-labeled Annexin V, and when late apoptosis or necrosis occurs, the cell membrane permeability increases, so that nucleic acid dye (PI) which would not enter normal cells can enter cells and stain DNA inside the cells, and early apoptosis, late apoptosis and necrotic cells can be identified and quantified by using Annexin V and PI double staining methods.
The measurement method is as follows:
(1) Tumor cells were inoculated into 5×5cm 2 culture dishes at a density of 30 ten thousand per dish, and incubated in a CO 2 incubator for 24 hours;
(2) When the cells to be tested enter the logarithmic growth phase, compound 6 with different concentrations (0 mu M, 0.5 mu M, 1.0 mu M and 3.0 mu M) is added for administration treatment;
(3) After 24h of drug administration treatment, the cells to be tested are transferred to a 15mL centrifuge tube, 1000g is centrifuged for 5min, the supernatant is discarded, the cells are collected, and PBS is used for cleaning for 2 times;
(4) 195 μl of diluted 1× Annexin V Binding Buffer resuspended cells were added to the cells;
(5) Sequentially adding 5 mu L of annexin V-FITC and 5 mu L of PI dye into the resuspended cells, and gently mixing;
(6) Incubating at 20deg.C in dark for 15min, blowing and mixing, and detecting on a machine after incubation.
The effect of compound 6 on apoptosis of triple negative breast cancer cells HCC1937 at various concentrations is shown in figure 17.
As can be seen from fig. 17, compound 6 was able to significantly induce apoptosis in triple negative breast cancer cells HCC1937, and after 24 hours of treatment with 3.0 μm, about 36.2% of triple negative breast cancer cells HCC1937 were early-phase apoptotic and 1.12% of triple negative breast cancer cells HCC1937 were late-phase apoptotic.
It should be noted that the above examples are only examples for clearly illustrating the present invention, and are not limiting to the embodiments of the present invention. Other variations or modifications of the above teachings will be apparent to those of ordinary skill in the art. Not all embodiments are exhaustive. All obvious changes or modifications which are obvious from the technical proposal of the invention are still within the protection scope of the invention.

Claims (10)

1. Pyrazolopyridine derivative is characterized in that the pyrazolopyridine derivative has the following structure:
、、
、、
、 And
。
2. The method for producing pyrazolopyridine derivative according to claim 1, comprising the steps of:
Step 1, cyclizing: dissolving 5-bromo-2-chloronicotinonitrile in absolute ethyl alcohol, heating to 85 ℃, dropwise adding hydrazine hydrate, maintaining the temperature of a reaction system, stirring, and after the reaction is finished, sequentially carrying out low-temperature recrystallization, vacuum filtration and drying to obtain a compound 2;
Step 2, carbon-carbon coupling: dissolving a compound 2, a compound A and potassium phosphate in dioxane aqueous solution, wherein the compound A is a compound 3, a compound 9, a compound 12 or a compound 15, blowing nitrogen, adding ditri-butyl phosphine palladium into a reaction system, reacting at 80-110 ℃, and sequentially carrying out diatomite vacuum filtration, ethyl acetate and water extraction and drying after the reaction is finished to obtain a compound B;
step 3, amidation: dissolving a compound B in N, N-dimethylformamide, adding a compound C, wherein the compound C is bromoacetyl chloride, chloroacetyl chloride or 2, 2-trichloroethyl chloroformate, reacting at 20 ℃, and sequentially quenching, extracting ethyl acetate and water, vacuum rotary evaporating, passing through a column and drying after the reaction is finished to obtain the pyrazolopyridine derivative of claim 1;
Wherein the structures of compound 2, compound 3, compound 9, compound 12 and compound 15 are shown below, respectively:
、、、、。
3. The method for producing pyrazolopyridine derivative according to claim 2, wherein in step 1, the ratio of 5-bromo-2-chloronicotinonitrile to hydrazine hydrate is 23mmol:115mmol.
4. The method for producing pyrazolopyridine derivative according to claim 2, wherein in step 2, the ratio of the amounts of compound 2, compound a, potassium phosphate and di-tri-t-butylphosphine palladium is 14mmol:16.8mmol:28mmol:0.7mmol.
5. The method for producing pyrazolopyridine derivative according to claim 2, wherein in step 3, the ratio of compound B to compound C is 0.3mmol:0.6mmol.
6. The method for producing pyrazolopyridine derivative according to claim 2, further comprising a methylation step, specifically:
After the step 2 is finished, the compound B is dissolved in N, N-dimethylformamide, sodium hydride is added under ice bath, then potassium iodide is added dropwise for reaction, quenching, ethyl acetate and water extraction, vacuum rotary evaporation, column passing and drying are sequentially carried out after the reaction is finished, the compound D is obtained, methylation is finished, and then the compound D is substituted for the compound B for amidation reaction.
7. The method for producing pyrazolopyridine derivative according to claim 6, wherein in the methylation step, the amount ratio of compound B, sodium hydride and potassium iodide is 0.3mmol:0.45mmol:0.45mmol.
8. The use of pyrazolopyridine derivative according to claim 1 for preparing an anti-breast cancer medicament, wherein the pyrazolopyridine derivative is compound 5, compound 6, compound 11, compound 17 or compound 19.
9. The use of pyrazolopyridine derivative according to claim 1 for preparing an anti-pancreatic cancer drug, wherein the pyrazolopyridine derivative is compound 5, compound 6, compound 7, compound 11, compound 14, compound 17 and compound 19.
10. The use of pyrazolopyridine derivative according to claim 1 in the preparation of anti-gastric cancer drugs, wherein the pyrazolopyridine derivative is compound 5, compound 6, compound 11, compound 14, compound 17 and compound 19.
CN202410559979.XA 2024-05-08 2024-05-08 Pyrazolopyridine derivative, and preparation method and application thereof Active CN118126038B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202410559979.XA CN118126038B (en) 2024-05-08 2024-05-08 Pyrazolopyridine derivative, and preparation method and application thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202410559979.XA CN118126038B (en) 2024-05-08 2024-05-08 Pyrazolopyridine derivative, and preparation method and application thereof

Publications (2)

Publication Number Publication Date
CN118126038A CN118126038A (en) 2024-06-04
CN118126038B true CN118126038B (en) 2024-07-12

Family

ID=91239260

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202410559979.XA Active CN118126038B (en) 2024-05-08 2024-05-08 Pyrazolopyridine derivative, and preparation method and application thereof

Country Status (1)

Country Link
CN (1) CN118126038B (en)

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20100057433A (en) * 2008-11-21 2010-05-31 한국화학연구원 Novel pyrazolo[4,3-b]pyridine derivatives or pharmaceutically acceptable salt thereof, preparation method thereof and pharmaceutical composition containing the same as an active ingredient
KR20140019055A (en) * 2012-07-16 2014-02-14 한국과학기술연구원 Pyrazolopyridine or indazole derivatives as protein kinase inhibitors

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2004509891A (en) * 2000-09-22 2004-04-02 スミスクライン　ビーチャム　パブリック　リミテッド　カンパニー Pyrazolopyridine and pyrazolopyridazine as antidiabetic drugs
US20090306045A1 (en) * 2005-12-22 2009-12-10 Ira Mellman Inhibition of Glycogen Synthase Kinase and Methods of Treating Autoimmune or Immune Inflammatory Disease
PE20090717A1 (en) * 2007-05-18 2009-07-18 Smithkline Beecham Corp QUINOLINE DERIVATIVES AS PI3 KINASE INHIBITORS
KR20120063283A (en) * 2010-12-07 2012-06-15 제일약품주식회사 Novel pyrazolopyridine derivatives or pharmaceutically acceptable salts thereof, process for the preparation thereof and pharmaceutical composition comprising the same
CN117820332B (en) * 2024-03-04 2024-05-14 烟台大学 Benzothiophene-thiourea compound and preparation method and application thereof

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20100057433A (en) * 2008-11-21 2010-05-31 한국화학연구원 Novel pyrazolo[4,3-b]pyridine derivatives or pharmaceutically acceptable salt thereof, preparation method thereof and pharmaceutical composition containing the same as an active ingredient
KR20140019055A (en) * 2012-07-16 2014-02-14 한국과학기술연구원 Pyrazolopyridine or indazole derivatives as protein kinase inhibitors

Also Published As

Publication number Publication date
CN118126038A (en) 2024-06-04

Similar Documents

Publication Publication Date Title
CN106220641B (en) Containing the indoles volution compound and the preparation method and application thereof for more creating blue hydrocarbon Azulene structure
CN112047950A (en) Imidazopyrazine derivative and synthesis method and application thereof
CN118126038B (en) Pyrazolopyridine derivative, and preparation method and application thereof
CN111362962B (en) Tetrafluorobenzyl norcantharidin carboxylate and synthesis method thereof
CN108484623B (en) Camptothecin derivative and preparation method and application thereof
CN106478692A (en) Copper-nitrate complex and its synthetic method and application with 1 (2 pyridine) 9 benzyl β carboline as part
CN110526854A (en) A kind of ɑ, alpha, beta-unsaturated ketone derivative, preparation method and the purposes as drug
CN106188075B (en) Indoles volution compound and the preparation method and application thereof
CN109942455A (en) Gossypol with anti-tumor activity-Eflornithine schiff base compounds and its synthetic method
CN111892596B (en) Preparation and application of beta-carboline compound containing polyfluoropyrazole structure
CN104230786B (en) Indole-structure-containing compound with anti-tumor activity and synthesis method thereof
CN111892594B (en) Preparation and application of 1- (3,4, 5-trimethoxyphenyl) -beta-carboline acylhydrazone containing substituted pyrazole unit
CN111961049B (en) Beta-carboline derivative containing 1, 3-dimethyl-5-aryloxy pyrazole and preparation method and application thereof
CN106478677B (en) The chlorination copper complex of 1 (2 pyridine) 9 (2 Phenoxyethyl) β carbolines and synthetic method and application
CN108586432B (en) 3- (indole-5-yl) -indazole derivative and application thereof
CN106478690A (en) The chlorination copper complex of 1 (2 pyridine) 9 (4 phenyl butyl) β carboline and its synthetic method and application
CN106478676B (en) The copper-nitrate complex of 1 (2 pyridine) 9 (2 Phenoxyethyl) β carbolines and synthetic method and application
CN109053732B (en) Para-fluorine substituted pyridazinone structure-containing spiro [ indolizine-pyrazoline ] derivative and preparation method and application thereof
CN109232572B (en) P-methyl-substituted pyridazinone-structure-containing spiro [ indolizine-pyrazoline ] derivative and preparation method and application thereof
CN106478678B (en) The copper-nitrate complex of 1 (2 pyridine) 9 (methyl of naphthalene 2) β carbolines and its synthetic method and application
CN106432288B (en) The copper-nitrate complex of 1 (2 pyridine) 9 (2 Benzyloxyethyl) β carbolines and synthetic method and application
CN106632415B (en) The chlorination copper complex of 1 (2 pyridine) 9 (methyl of naphthalene 2) β carbolines and its synthetic method and application
CN106478675B (en) The chlorination copper complex of 1 (2 pyridine) 9 (2 Benzyloxyethyl) β carbolines and synthetic method and application
CN106478679B (en) The copper-nitrate complex of 1 (2 pyridine) 9 Cvclopropvlmethvl β carbolines and its synthetic method and application
CN106632423B (en) The copper-nitrate complex and its synthetic method of 1- (2- pyridines) -9- (2- phenylethyls)-B-carboline and application

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
GR01 Patent grant
GR01 Patent grant