CN113321651B

CN113321651B - Pyrazolopyridine hydroxamic acid compound, and preparation method and application thereof

Info

Publication number: CN113321651B
Application number: CN202110692076.5A
Authority: CN
Inventors: 师健友
Original assignee: Sichuan Peoples Hospital of Sichuan Academy of Medical Sciences
Current assignee: Sichuan Peoples Hospital of Sichuan Academy of Medical Sciences
Priority date: 2021-06-22
Filing date: 2021-06-22
Publication date: 2022-06-14
Anticipated expiration: 2041-06-22
Also published as: CN113321651A

Abstract

The invention relates to the technical field of chemical synthesis, and particularly relates to a pyrazolopyridine hydroxamic acid compound, and a preparation method and application thereof. The pyrazolopyridine hydroxamic acid compound comprises a compound shown in a formula (1), an isomer thereof, a hydrate thereof or a pharmaceutically acceptable salt thereof,

Description

Pyrazolopyridine hydroxamic acid compound, and preparation method and application thereof

Technical Field

The invention relates to the technical field of chemical synthesis, and particularly relates to a pyrazolopyridine hydroxamic acid compound, and a preparation method and application thereof.

Background

The traditional antitumor drug treatment has the defects of large toxic and side effects, low selectivity, multi-drug resistance and the like, so that the small-molecule targeted therapeutic drug becomes a research hotspot of the antitumor drugs in recent years. In recent years, more and more studies have shown that Histone Deacetylases (HDACs) are closely related to the development of tumors. Histones play important roles in gene expression, cell growth, differentiation and apoptosis in vivo by participating in structural modification of chromatin and in regulation of other proteins.

The hydroxamic acid compound is used as a histone deacetylase inhibitor, a matrix metalloproteinase inhibitor, a 5-lipoxygenase inhibitor and the like in medicine to play an anti-tumor effect, and the hydroxamic acid compound and derivatives thereof have strong physiological and pharmacological activities, so that the hydroxamic acid compound and derivatives thereof with novel structures are designed and a synthesis method thereof is developed, and the hydroxamic acid compound and derivatives thereof have important significance in the fields of synthetic chemistry, pharmaceutical chemistry, pharmacology and the like.

In view of this, the invention is particularly proposed.

Disclosure of Invention

The invention aims to provide a pyrazolopyridine hydroxamic acid compound, a preparation method and application thereof. Disclosed is a novel compound which can inhibit the proliferation of cancer cells well and has an excellent therapeutic effect on tumors.

The invention is realized by the following steps:

in a first aspect, the present invention provides a pyrazolopyridine hydroxamic acid compound comprising a compound represented by formula (1), an isomer thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof,

wherein n is an integer, and R is any one of a substituted or unsubstituted aromatic ring group and a substituted alkyl group.

In a second aspect, the present invention provides a method for preparing pyrazolopyridine hydroxamic acid compounds according to the above embodiments, wherein the pyrazolopyridine hydroxamic acid compounds are synthesized according to the following synthetic route,

wherein X is halogen.

In a third aspect, the present invention provides a use of the pyrazolopyridine hydroxamic acid compound according to the previous embodiment in the preparation of a medicament for treating any one of leukemia, lung small cell lung cancer, liver cancer, stomach cancer, prostate cancer, pancreatic cancer, ovarian cancer, bladder cancer, kidney cancer, melanoma, breast cancer, myeloma, lymphoma, esophageal cancer, rectal cancer, thyroid cancer, cervical cancer, and colon cancer.

The invention has the following beneficial effects: the embodiment of the invention forms pyrazolopyridine into the hydroxamic acid compound, so that the pyrazolopyridine hydroxamic acid compound can well inhibit cancer cell proliferation, and further has an excellent treatment effect on tumors.

Detailed Description

In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are conventional products which are not indicated by manufacturers and are commercially available.

The invention provides a pyrazolopyridine hydroxamic acid compound, which comprises a compound shown as a formula (1), an isomer, a hydrate or a pharmaceutically acceptable salt thereof,

wherein n is an integer and R is a substituted or unsubstituted aromatic ring group.

Specifically, n is 1 to 20, preferably 1 to 10, more preferably 5 or 6. N may be any integer such as 1, 2, 3, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, in addition to 5 or 6.

Further, R is an unsubstituted unfused aromatic or heteroaromatic group, for example a thiophene group or a thiazole group, in particular

The unsubstituted unfused aromatic ring group or the aromatic heterocyclic group may be any of unsubstituted unfused aromatic ring groups which can be grafted onto the parent nucleus of formula (1) such as a benzene ring, a pyran ring, or a piperazine ring group, and which have a good antitumor effect.

Further, R is an unsubstituted or substituted fused aromatic or heteroaromatic group, for example

Further, R is a substituted-unfused aromatic ring group or aromatic heterocyclic group, such as a substituted benzene ring, more preferably a mono-substituted benzene ring or a di-substituted benzene ring; wherein, the mono-substituted benzene ring is an ortho-substituted benzene ring, a meta-substituted benzene ring or a para-substituted benzene ring; the disubstituted benzene ring is a3, 4-substituted benzene ring; the disubstituted benzene ring may be a3, 5-substituted benzene ring, a 2, 4-substituted benzene ring or a 2, 5-substituted benzene ring, and the substituted benzene ring may be a trisubstituted benzene ring, a tetrasubstituted benzene ring or other polysubstituted benzene rings.

Further, the substituent of the substituted benzene ring is selected from alkyl or amino; wherein the amine group is a tertiary amine group, more preferably

Wherein R is₁And R₂Are each an alkyl group; alkyl of substituted benzene ringRadical, R₁And R₂Are respectively and independently C1-C10 alkyl, preferably C1-C10 straight-chain alkyl or C3-C10 branched-chain alkyl, more preferably C1-C4 straight-chain alkyl or C4-C8 branched-chain alkyl, and more preferably methyl, n-butyl or tert-butyl.

R may also be a substituted alkyl group such as, for example,

wherein R is₃Is halogen (e.g., bromine, chlorine, iodine, etc.) or alkyl (e.g., methyl, ethyl, propyl, t-butyl, n-butyl, etc.), but may of course also be

n is 1 to 5.

Specifically, the pyrazolopyridine hydroxamic acid compound is selected from any one of the compounds shown in the following structural formula:

this example also provides a method for preparing the pyrazolopyridine hydroxamic acid compound, specifically, the pyrazolopyridine hydroxamic acid compound is synthesized by referring to the following synthetic route,

wherein X is halogen, e.g. bromine or chlorine, etc., and X₂Or PX₅May be a different halogen, e.g. X₂Is Br₂And PX₅Is PCl₅. Wherein, when compound 7 is formed: the mol ratio of the compound 5 to the compound 6 to the condensing agent to the base is 1:1-1.2:2.2-2.5: 1.8-2.2. To form compound 9: the molar ratio of the compound 7 to the compound 8 is 1: 1.2-1.5. The above molar ratio is further favorable for the progress of each reaction and the formation of the most favorable product. Wherein the condensing agent can be HATU, or other condensation in the prior art, and the alkali can be TEA, or other alkaline substances in the prior art.

Furthermore, the embodiment of the present invention also provides an application of the pyrazolopyridine hydroxamic acid compound in the preparation of a medicament for treating any one of leukemia, lung small cell lung cancer, liver cancer, stomach cancer, prostate cancer, pancreatic cancer, ovarian cancer, bladder cancer, kidney cancer, melanoma, breast cancer, myeloma, lymphoma, esophageal cancer, rectal cancer, thyroid cancer, cervical cancer, colon cancer and other cancers. The pyrazolopyridine hydroxamic acid compound has excellent capacity of inhibiting cancer cell proliferation, has excellent activity of inhibiting cancer cell proliferation on human chronic myelogenous leukemia cells (K562), cervical cancer cells (Hela), human colon cancer cells (HCT116), non-small cell lung cancer cells (A549), melanoma cells (A375), lymphoma cells (Raji), breast cancer cells (MCF-7), liver cancer cells (HepG2), prostate cancer cells (PC-3) and pancreatic cancer cells (SW1990), and further has a good treatment effect on the diseases.

The features and properties of the present invention are described in further detail below with reference to examples.

Example 1

This example provides pyrazolopyridine hydroxamic acids (nos. 5-71) of the formula:

the present embodiment provides a preparation method of 5 to 71, which includes:

s1, synthesis of compound 2 by reference to the following synthesis route;

20 g of 2-chloro-3-cyanopyridine (144.3 mmol) were dissolved in about 60 ml of glacial acetic acid and reacted under reflux for about 45 h. And (4) tracking and monitoring by thin-layer chromatography, and after the reaction is completed, a large amount of white solid is separated out after the reaction liquid is cooled. Filtration, washing of the precipitate with a small amount of glacial acetic acid and drying gave compound 2 in 60% yield.

S2, synthesis of compound 3 by reference to the following synthesis route;

5 g of 2-hydroxy-3-cyanopyridine (1.0 eq, 41.6 mmol) was dissolved in glacial acetic acid, 3.2 ml of liquid bromine (1.5 eq, 62.4 mmol) was slowly added dropwise to the reaction mixture and allowed to react at room temperature (25 ℃) for about 2h (the inventors found that the reaction was achieved at 0.5-24 h). And (5) carrying out tracking monitoring by thin layer chromatography, and adding 10% sodium sulfite solution into the reaction liquid to quench excessive liquid bromine after the reaction is completed. 100 ml of water was added, extracted with ethyl acetate (50 ml. times.3), the organic layers were combined and washed with saturated brine, dried over anhydrous magnesium sulfate for half an hour, and the solvent was recovered to obtain compound 3 in 85% yield.

S3, synthesis of compound 4 by reference to the following synthesis route;

5 g of 2-hydroxy-3-cyano-5-bromopyridine (1 equivalent, 25 mmol) are added to 50 ml of phosphorus oxychloride, and 6.2 g of phosphorus pentachloride (1.2 equivalent, 30 mmol) are added in portions to the reaction mixture under reflux. And (3) tracking and monitoring by thin-layer chromatography, cooling the reaction liquid to room temperature after the reaction is completed, adding a small amount of the reaction liquid into a large amount of ice water for many times, stirring, adjusting the pH value to be neutral by using sodium hydroxide, and separating out a white precipitate. Filtration, washing of the precipitate with water and drying gave compound 4 in 52% yield.

S4, synthesis of compound 5 by reference to the following synthesis route;

3.2 g of 2-chloro-3-cyano-5-bromopyridine (1 eq, 14.7 mmol) and 2.15 ml of hydrazine hydrate (3 eq, 44.2 mmol) are dissolved in ethanol and reacted under reflux. And (3) tracking and monitoring by thin-layer chromatography, after the reaction is completed, spin-drying the reaction solution, adding a proper amount of water into a reaction bottle for ultrasonic treatment, filtering the obtained yellow precipitate, and washing with water to obtain a compound 5 with the yield of 83%.

S5 Synthesis of Compound 7 by reference to the following Synthesis route;

1 equivalent of compound 5 and 1.2 equivalents of compound 6 were dissolved in N, N-dimethylformamide, and 2.5 equivalents of HATU and 2 equivalents of triethylamine were added thereto, followed by reaction at room temperature (about 25 ℃ C.). And (5) tracking and monitoring by thin-layer chromatography, and adding a large amount of ice water into the reaction solution after the reaction is completed to precipitate and separate out. Filtration, washing of the precipitate with water and drying gave compound 7 in 78% yield.

S6, synthesis of compound 9 by reference to the following synthesis route;

1 equivalent of Compound 7 and 1.5 equivalents of Compound 8 were dissolved in acetonitrile, and 3 equivalents of cesium carbonate were added to conduct reaction at ordinary temperature. Monitoring by thin layer chromatography, after the reaction is completed, spin-drying the reaction solution, adding water and ethyl acetate for extraction, combining organic layers, washing with saturated saline solution, drying over anhydrous magnesium sulfate for half an hour, and recovering the solvent to obtain compound 9 with a yield of 63%.

S7, synthesizing pyrazolopyridine hydroxamic acid compounds by referring to the following synthetic route;

dissolving 1 equivalent of compound 9 in methanol, adding 10 equivalents of compound 10 and 5 equivalents of sodium hydroxide under the condition of ice bath at 0 ℃, and standing for 5 minutes for reaction at normal temperature. And (4) tracking and monitoring by thin layer chromatography, after the reaction is completed, spin-drying the reaction solution, adjusting the pH to 6-7 by using dilute hydrochloric acid, and precipitating. Filtration, washing of the precipitate with water and drying gave compound 11, i.e. 5-71, in 52% yield.

Pyrazolopyridine hydroxamic acid compounds were characterized according to the following characterization data:¹H NMR(400MHz,DMSO-d₆)δ11.35(s,1H),10.31(s,1H),8.65(s,1H),7.92(d,J＝5.5Hz,1H),6.82(s,1H),6.73(s,1H),6.05(s,1H),3.15(s,2H),2.06–1.83(m,2H),1.67–1.07(m,6H).

HRMS(ESI)m/z calcd for C₁₇H₂₁N₇O₃+H372.1785,found 372.1782.

example 2

This example provides pyrazolopyridine hydroxamic acids (nos. 5-72) of the formula:

the synthesis method is the same as that of example 1, except that: replacement of Compound 6 of example 1 with

The characterization data are as follows:¹H NMR(400MHz,DMSO-d₆)δ10.34(s,1H),8.66(s,1H),8.10(t,J＝5.9Hz,1H),7.19(d,J＝6.1Hz,1H),7.05–6.98(m,2H),3.18(dd,J＝8.2,5.9Hz,2H),2.28(d,J＝4.5Hz,6H),1.99–1.90(m,2H),1.49(dp,J＝14.6,7.5Hz,4H),1.28(p,J＝7.2,6.7Hz,2H).

HRMS(ESI)m/z calcd for C₂₁H₂₆N₆O₃+H411.2145,found 411.2154.

example 3

This example provides pyrazolopyridine hydroxamic acids (nos. 5-74) of the formula:

The characterization data are as follows:¹H NMR(400MHz,DMSO-d₆)δ10.38(s,1H),8.59(s,1H),8.33(s,1H),7.74(d,J＝8.1Hz,2H),7.26(d,J＝8.1Hz,2H),3.22(q,J＝6.7Hz,2H),2.59(s,2H),2.00–1.89(m,2H),1.64–1.43(m,6H),1.38–1.19(m,4H),0.91(s,3H).

HRMS(ESI)m/z calcd for C₂₃H₃₀N₆O₃+H 439.2458,found 439.2454.

example 4

This example provides pyrazolopyridine hydroxamic acids (nos. 5-77) having the following structural formula:

The characterization data are as follows:¹H NMR(400MHz,DMSO-d₆)δ10.35(s,1H),8.66(s,1H),8.47(t,J＝5.4Hz,1H),7.74(dd,J＝9.6,4.1Hz,2H),7.21–7.03(m,1H),3.21(q,J＝6.8Hz,2H),2.03–1.87(m,2H),1.51(h,J＝8.4Hz,4H),1.27(p,J＝7.1,6.0Hz,2H).

HRMS(ESI)m/z calcd for C₁₇H₂₀N₆O₃S+H389.1397,found 389.1395.

example 5

This example provides pyrazolopyridine hydroxamic acids (nos. 5-710) having the following structural formula:

The characterization data are as follows:¹H NMR(400MHz,DMSO-d₆)δ10.35(s,1H),8.66(s,1H),8.08(t,J＝5.5Hz,1H),7.71(d,J＝8.8Hz,2H),6.69(d,J＝8.9Hz,2H),3.20(q,J＝7.6,7.1Hz,2H),2.96(s,6H),2.01–1.90(m,2H),1.50(h,J＝7.7,7.3Hz,4H),1.35–1.20(m,2H).

HRMS(ESI)m/z calcd for C₂₁H₂₇N₇O₃+H 426.2254,found 426.2252.

example 6

This example provides pyrazolopyridine hydroxamic acids (nos. 5-712) of the formula:

The characterization data are as follows:¹H NMR(400MHz,DMSO-d₆)δ10.33(s,1H),8.66(s,1H),8.35(t,J＝5.5Hz,1H),7.87–7.68(m,2H),7.46(d,J＝8.4Hz,2H),3.23(q,J＝6.9Hz,2H),2.02–1.85(m,2H),1.59–1.43(m,4H),1.32–1.21(m,11H).

HRMS(ESI)m/z calcd for C₂₃H₃₀N₆O₃+H 439.2458,found 439.2466.

example 7

This example provides pyrazolopyridine hydroxamic acids (nos. 5-715) of the formula:

The characterization data are as follows:¹H NMR(400MHz,DMSO-d₆)δ10.35(s,1H),8.67(s,1H),8.29(t,J＝5.5Hz,1H),7.43(d,J＝8.0Hz,2H),6.98(d,J＝8.2Hz,1H),4.06(qd,J＝6.9,3.7Hz,4H),3.21(q,J＝6.7Hz,2H),1.95(t,J＝7.4Hz,2H),1.51(h,J＝7.0Hz,4H),1.41–1.17(m,9H).

HRMS(ESI)m/z calcd for C₂₃H₃₀N₆O₅+H 471.2357,found 471.2353.

example 8

This example provides pyrazolopyridine hydroxamic acids (nos. 5-716) of the formula:

The characterization data are as follows:¹H NMR(400MHz,DMSO-d₆)δ10.38(s,1H),8.90–8.36(m,2H),7.63(d,J＝4.0Hz,1H),7.17(d,J＝4.0Hz,1H),2.11–1.85(m,2H),1.49(dq,J＝14.2,7.1Hz,3H),1.37–1.10(m,5H).

HRMS(ESI)m/z calcd for C₁₇H₁₉ClN₆O₃S+H 423.1007,found 423.1015.

example 9

This examplePyrazolopyridine hydroxamic acids (numbered 5-718) are provided having the following structural formula:

The characterization data are as follows:¹H NMR(400MHz,DMSO-d₆)δ13.56(s,1H),10.41(s,1H),8.58(s,2H),6.66(s,1H),3.85(s,3H),3.32(s,4H),2.57–2.44(m,6H),2.33(s,3H).

HRMS(ESI)m/z calcd for C₁₈H₂₄N₈O₃+H 401.2050,found 401.2046.

example 10

This example provides pyrazolopyridine hydroxamic acids (nos. 5-720) having the following structural formula:

the synthesis method is the same as that of example 1, except that: substitution of Compound 6 of example 1

Is composed of

The characterization data are as follows:¹H NMR(400MHz,DMSO-d₆)δ10.35(s,1H),8.65(s,1H),8.43–8.24(m,1H),7.51(d,J＝8.4Hz,1H),7.02–6.85(m,2H),3.77(s,3H),1.95(t,J＝6.9Hz,2H),1.50(h,J＝6.9Hz,4H),1.39–1.07(m,4H).

HRMS(ESI)m/z calcd for C₂₀H₂₃BrN₆O₄+H 491.1043,found 491.1036.

example 11

This example provides pyrazolopyridine hydroxamic acids (nos. 5-721) of the formula:

the synthesis method is the same as in example 1, except that: replacement of Compound 6 of example 1 with

The characterization data are as follows:¹H NMR(400MHz,DMSO-d₆)δ10.34(s,1H),8.67(s,1H),8.13–7.94(m,1H),7.67(d,J＝8.9Hz,2H),6.63(d,J＝9.0Hz,2H),3.19(dd,J＝13.1,5.1Hz,4H),1.95(t,J＝7.4Hz,2H),1.50(dp,J＝15.2,8.0,7.5Hz,4H),1.26(dt,J＝12.1,6.1Hz,3H),1.10(t,J＝7.0Hz,7H).

HRMS(ESI)m/z calcd for C₂₃H₃₁N₇O₃+H 454.2567,found 454.2567.

example 12

This example provides pyrazolopyridine hydroxamic acids (nos. 5-724) having the following structural formula:

The characterization data are as follows:¹H NMR(400MHz,DMSO-d₆)δ10.42(s,1H),8.57(t,J＝5.5Hz,1H),8.11(s,1H),7.89–7.76(m,2H),7.77–7.67(m,2H),7.53(dt,J＝18.9,7.8Hz,3H),7.41(t,J＝7.3Hz,1H),3.17(s,2H),1.96(t,J＝7.4Hz,2H),1.54(dq,J＝15.0,7.6Hz,4H),1.30(p,J＝9.2,8.7Hz,2H).

HRMS(ESI)m/z calcd for C₂₅H₂₆N₆O₃+H 459.2145,found 459.2153.

example 13

This example provides pyrazolopyridine hydroxamic acids (nos. 5-726) of the formula:

The characterization data are as follows:¹H NMR(400MHz,DMSO-d₆)δ8.57–8.46(m,1H),7.78(d,J＝8.5Hz,2H),7.67(d,J＝8.5Hz,2H),3.23(q,J＝6.8Hz,2H),3.17(d,J＝4.0Hz,2H),1.95(t,J＝7.4Hz,2H),1.51(p,J＝8.1,7.1Hz,3H),1.27(dt,J＝14.6,7.3Hz,2H).

HRMS(ESI)m/z calcd for C₁₉H₂₁BrN₆O₃+H 461.0938,found 461.0944.

example 14

This example provides pyrazolopyridine hydroxamic acid compounds (nos. 5-727) having the following structural formula:

The characterization data are as follows:¹H NMR(400MHz,DMSO-d₆)δ12.18(s,1H),10.33(s,1H),8.66(s,1H),8.43–8.35(m,1H),8.09–7.97(m,1H),7.41–7.21(m,4H),5.66(s,1H),3.39(s,2H),3.02(q,J＝6.7Hz,2H),1.93(t,J＝7.4Hz,2H),1.55–1.32(m,4H),1.30–1.13(m,2H).

HRMS(ESI)m/z calcd for C₂₀H₂₃ClN₆O₃+H 431.1599,found 431.1596.

example 15

This example provides pyrazolopyridine hydroxamic acids (nos. 6-73) of the formula:

Replacement of Compound 8 with

The characterization data are as follows:¹H NMR(400MHz,DMSO-d₆)δ10.33(s,1H),8.65(s,1H),8.35–8.18(m,1H),7.41(d,J＝24.9Hz,2H),6.98(s,1H),6.09(s,2H),3.27–3.13(m,2H),1.94(t,J＝7.3Hz,2H),1.48(q,J＝7.0Hz,4H),1.40–1.13(m,4H).

HRMS(ESI)m/z calcd for C₂₁H₂₄N₆O₅+H441.1887,found 441.1893.

example 16

This example provides pyrazolopyridine hydroxamic acids (nos. 6-74) of the formula:

the synthesis method is the same as that of example 15, except that: replacement of Compound 6 of example 15 with

The characterization data are as follows:¹H NMR(400MHz,DMSO-d₆)δ10.33(s,1H),8.70(s,1H),8.35(d,J＝8.7Hz,1H),7.74(d,J＝8.1Hz,2H),7.26(d,J＝8.1Hz,2H),3.26–3.13(m,3H),2.62(t,J＝7.8Hz,2H),1.99–1.90(m,2H),1.62–1.40(m,6H),1.37–1.21(m,6H),0.89(t,J＝7.6Hz,3H).

HRMS(ESI)m/z calcd for C₂₄H₃₂N₆O₃+H453.2615,found 453.2608.

example 17

This example provides pyrazolopyridine hydroxamates (numbered 6-77) having the following structural formula:

The characterization data are as follows:¹H NMR(400MHz,DMSO-d₆)δ10.33(s,1H),8.66(s,1H),8.44(t,J＝5.5Hz,1H),7.73(t,J＝3.7Hz,2H),7.13(dd,J＝4.8,3.8Hz,1H),3.21(q,J＝6.7Hz,2H),1.94(t,J＝7.3Hz,2H),1.65–1.39(m,4H),1.38–1.13(m,4H).

HRMS(ESI)m/z calcd for C₁₈H₂₂N₆O₃S+H 403.1553,found 403.1559.

example 18

This example provides pyrazolopyridine hydroxamic acids (nos. 6-78) of the formula:

The characterization data are as follows:¹¹H NMR(400MHz,DMSO-d₆)δ10.34(s,1H),8.66(s,1H),8.06(t,J＝5.2Hz,1H),7.73(dd,J＝7.7,1.8Hz,1H),7.43(ddd,J＝8.8,7.3,1.9Hz,1H),7.17–7.07(m,1H),7.01(td,J＝7.5,1.0Hz,1H),4.06(t,J＝6.4Hz,2H),3.28(q,J＝6.7Hz,2H),2.02–1.86(m,2H),1.87–1.71(m,2H),1.64–1.40(m,4H),1.30(ddt,J＝20.5,14.3,7.1Hz,4H),1.01(t,J＝7.4Hz,3H).

HRMS(ESI)m/z calcd for C₂₃H₃₀N₆O₄+H 455.2408,found 455.2404.

example 19

This example provides pyrazolopyridine hydroxamates (Nos. 6-71)0) The structural formula is as follows:

The characterization data are as follows:¹H NMR(400MHz,DMSO-d₆)δ10.31(s,1H),8.65(s,1H),8.17–7.89(m,1H),7.71(d,J＝8.3Hz,2H),6.69(d,J＝8.9Hz,2H),3.20(q,J＝6.5Hz,2H),2.96(s,6H),1.94(t,J＝7.3Hz,2H),1.57–1.39(m,4H),1.37–1.18(m,4H).

HRMS(ESI)m/z calcd for C₂₂H₂₉N₇O₃+H440.2426,found 440.2423.

example 20

This example provides pyrazolopyridine hydroxamic acids (nos. 6-712) of the formula:

The characterization data are as follows:¹H NMR(400MHz,DMSO-d₆)δ10.33(s,1H),8.66(s,1H),8.34(t,J＝5.5Hz,1H),7.76(d,J＝8.4Hz,2H),7.46(d,J＝8.4Hz,2H),3.23(d,J＝6.4Hz,2H),1.94(t,J＝7.4Hz,2H),1.55–1.43(m,4H),1.27(s,13H).HRMS(ESI)m/z calcd for C₂₄H₃₂N₆O₃+Na475.2433,found 475.2440.

example 21

This example provides pyrazolopyridine hydroxamic acids (nos. 6-715) of the formula:

The characterization data are as follows:¹H NMR(400MHz,DMSO-d₆)δ10.33(s,1H),8.65(s,1H),8.27(t,J＝5.5Hz,1H),7.43(d,J＝8.2Hz,2H),6.98(d,J＝8.2Hz,1H),4.06(qd,J＝6.8,3.9Hz,4H),3.21(q,J＝6.9Hz,2H),1.94(t,J＝7.3Hz,2H),1.61–1.40(m,4H),1.40–1.15(m,10H).

HRMS(ESI)m/z calcd for C₂₄H₃₂N₆O₅+Na 507.2331,found 507.2333.

example 22

This example provides pyrazolopyridine hydroxamic acids (nos. 6-721) of the formula:

The characterization data are as follows:¹H NMR(400MHz,DMSO-d₆)δ10.37(s,1H),8.67(s,1H),8.00(t,J＝5.2Hz,1H),7.67(d,J＝8.8Hz,2H),6.63(d,J＝8.6Hz,2H),3.19(d,J＝10.4Hz,3H),2.51(s,3H),1.94(t,J＝7.3Hz,2H),1.59–1.39(m,4H),1.37–1.20(m,4H),1.10(t,J＝6.9Hz,6H).

HRMS(ESI)m/z calcd for C₂₄H₃₃N₇O₃+Na 490.2542,found 490.2548.

example 23

This example provides pyrazolopyridine hydroxamic acids (nos. 6-722) of the formula:

The characterization data are as follows:¹H NMR(400MHz,DMSO-d₆)δ11.17(s,1H),10.30(s,1H),8.56(dd,J＝12.7,2.2Hz,1H),8.04(d,J＝3.7Hz,1H),7.48(d,J＝8.3Hz,2H),7.20(d,J＝8.3Hz,2H),3.75(s,1H),3.18(s,2H),3.02(s,2H),1.99–1.84(m,2H),1.58–1.29(m,4H),1.22(s,4H).

HRMS(ESI)m/z calcd for C₂₁H₂₅BrN₆O₃+H 489.1251,found 489.1248.

example 24

This example provides pyrazolopyridine hydroxamic acids (nos. 6-724) having the following structural formula:

The characterization data are as follows:¹H NMR(400MHz,DMSO-d₆)δ8.58(t,J＝5.5Hz,1H),8.12(s,1H),7.82(dd,J＝12.4,7.9Hz,2H),7.77–7.66(m,2H),7.53(dt,J＝18.9,7.8Hz,3H),7.40(t,J＝7.3Hz,1H),3.18(s,2H),1.95(t,J＝7.3Hz,2H),1.52(dt,J＝14.1,7.7Hz,4H),1.40–1.16(m,4H).

HRMS(ESI)m/z calcd for C₂₆H₂₈N₆O₃+Na 495.2120,found 495.2122.

example 25

This example provides pyrazolopyridine hydroxamic acids (nos. 6-729) of the formula:

The characterization data are as follows:¹H NMR(400MHz,DMSO-d₆)δ10.36(s,1H),8.63(d,J＝38.3Hz,1H),8.25(t,J＝5.4Hz,1H),7.47–7.09(m,2H),6.90(d,J＝8.2Hz,1H),4.27(q,J＝4.8Hz,4H),3.20(q,J＝6.7Hz,2H),1.94(t,J＝7.3Hz,2H),1.62–1.39(m,4H),1.40–1.14(m,4H).

HRMS(ESI)m/z calcd for C₂₂H₂₆N₆O₅+H 455.2044,found 455.2047.

activity study

Experimental Material

K562 cells (human chronic myelogenous leukemia cells), Hela cells (human cervical cancer cells) and HCT116 cells (human colon cancer cells), A549 (non-small cell lung cancer cells), A375 (melanoma cells), Raji (lymphoma cells), MCF-7 (breast cancer cells), HepG2 (liver cancer cells), PC-3 (prostate cancer cells), SW1990 (pancreatic cancer cells), DMEM medium, RPIMI1640 medium, extra-fetal bovine serum, PBS buffer, CCK-8, trypsin, sterile filter.

Collecting logarithmic phase cells, adjusting cell suspension concentration to 1 × 10⁴-2×10⁴Cell/ml (different cell concentrations were prepared depending on the cell type and growth rate). The cells were inoculated in 96-well plates at 100. mu.l per well, incubated overnight, and treated the next day with different concentrations of test compound (concentration gradient 80, 40, 20, 10, 5, 2.5, 1.25, 0.625. mu.M) at 100. mu.l per well in 6 duplicate wells. Setting solvent control group and blank control group, and selecting SAHA as positive control compound. After culturing at 37 ℃ for 48 hours in a 5% carbon dioxide incubator, 20. mu.l of CCK8 solution was added to each well, and the culture was continued for 4 hours, followed by measuring the absorbance using a microplate reader (. lamda. 450 nm). The average value of 6 multiple wells of each group of drug concentration is calculated according to the formula of inhibition rate: inhibition (%) - (control absorbance-experimental absorbance)/(control absorbance)Value-blank absorbance value) × 100%, IC50 values were calculated.

See table below for results.

As can be seen from the above tables, the compounds provided in the examples of the present invention have excellent proliferation inhibitory effects on leukemia cells, cervical cancer cells, colon cancer cells, non-small cell lung cancer cells, melanoma cells, lymphoma cells, breast cancer cells, liver cancer cells, prostate cancer cells, and pancreatic cancer cells, and can be used for the treatment of tumors. In addition, the experimental results also show that the compound provided by the embodiment of the invention also has similar inhibition effects on the proliferation of other cancer cells (such as cancer cells including gastric cancer cells, ovarian cancer cells, bladder cancer cells, renal cancer cells, myeloma cells, esophageal cancer cells, rectal cancer cells, thyroid cancer cells and the like), and further shows that the compound provided by the embodiment of the invention also has a certain therapeutic effect on other cancers.

In vitro HDAC inhibitory Activity assay

In vitro HDAC inhibitory activity assays were performed using fluorescence assays. First, a substrate is added to a sample of HDACs, which allows for deacetylation of the substrate and activation of the substrate. The substrate is then hydrolysed with Trypsin, which produces AMC groups. Finally, by measuring the fluorescence intensity of AMC group, we can calculate the inhibition rate of the compound to enzyme according to the fluorescence intensity, as shown in the following table.

Inhibition of HDACs by Compounds at a concentration of 1. mu.M

Two compounds with better activity were subjected to selective assay for HDAC isoforms, IC₅₀The (nM) results were as follows:

the above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A pyrazolopyridine hydroxamic acid compound characterized by comprising a compound represented by the formula (1) or a pharmaceutically acceptable salt thereof,

wherein n is 1-20, R is

And a substituted benzene ring, wherein,

n in (1) to (5);

the substituents of the substituted benzene ring are selected from

Or C1-C10 alkyl;

R₁and R₂Each independently is C1-C10 alkyl;

R₃is any one of halogen, methyl, ethyl, propyl, tertiary butyl and n-butyl.

2. The pyrazolopyridine hydroxamic acid compound according to claim 1, wherein n in formula (1) is 1 to 10.

3. A pyrazolopyridine hydroxamic acid compound according to claim 1, wherein n in formula (1) is 5 or 6.

4. A pyrazolopyridine hydroxamic acid compound according to claim 1, wherein R is a mono-substituted benzene ring or a di-substituted benzene ring.

5. The pyrazolopyridine hydroxamic acid compound according to claim 4, wherein the mono-substituted benzene ring is an ortho-substituted benzene ring, a meta-substituted benzene ring or a para-substituted benzene ring; the disubstituted benzene ring is a3, 4-substituted benzene ring.

6. Pyrazolopyridine hydroxamic acids according to claim 1, wherein alkyl, R of the substituted benzene ring₁And R₂Are respectively and independently C1-C10 straight-chain alkyl or C3-C10 branched-chain alkyl.

7. A pyrazolopyridine hydroxamic acid compound according to claim 6, wherein said alkyl group of substituted benzene ring, R₁And R₂Are respectively and independently C1-C4 straight-chain alkyl or C4-C8 branched-chain alkyl.

8. Pyrazolopyridine hydroxamic acids according to claim 1, wherein alkyl, R of the substituted benzene ring₁And R₂Each independently is methyl, n-butyl or tert-butyl.

9. A pyrazolopyridine hydroxamic acid compound, wherein the pyrazolopyridine hydroxamic acid compound is selected from any one of compounds represented by the following structural formulas:

10. a process for preparing pyrazolopyridine hydroxamic acid compound according to claim 1, characterized in that the pyrazolopyridine hydroxamic acid compound is synthesized by the following synthetic route,

wherein X is halogen, PX₅Wherein X is halogen, the condensing agent is HATU, the base is TEA, the catalyst used for forming the compound 9 by using the compound 7 and the compound 8 is cesium carbonate, and the compound 1 reacts with glacial acetic acid to form the compound 2.

11. The method of claim 10, wherein when compound 7 is formed: the molar ratio of the compound 5 to the compound 6 to the condensing agent to the base is 1:1-1.2:2.2-2.5: 1.8-2.2.

12. The method of claim 10, wherein, when compound 9 is formed: the molar ratio of the compound 7 to the compound 8 is 1: 1.2-1.5.

13. Use of the pyrazolopyridine hydroxamic acid compound according to claim 1 in the preparation of a medicament for the treatment of any one of leukemia, lung small cell lung cancer, liver cancer, stomach cancer, prostate cancer, pancreatic cancer, breast cancer, cervical cancer and human colon cancer.