CN114133390B - Harmine derivative as well as preparation method and application thereof - Google Patents
Harmine derivative as well as preparation method and application thereof Download PDFInfo
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- RAYHOJDYVGUYQQ-UHFFFAOYSA-N 7-methoxy-1-methyl-2,3,4,9-tetrahydro-1h-pyrido[3,4-b]indol-8-amine;hydrochloride Chemical compound [Cl-].C1CNC(C)C2=C1C1=CC=C(OC)C([NH3+])=C1N2 RAYHOJDYVGUYQQ-UHFFFAOYSA-N 0.000 title claims abstract description 28
- 238000002360 preparation method Methods 0.000 title abstract description 9
- BXNJHAXVSOCGBA-UHFFFAOYSA-N Harmine Chemical compound N1=CC=C2C3=CC=C(OC)C=C3NC2=C1C BXNJHAXVSOCGBA-UHFFFAOYSA-N 0.000 claims abstract description 55
- RERZNCLIYCABFS-UHFFFAOYSA-N Harmaline hydrochloride Natural products C1CN=C(C)C2=C1C1=CC=C(OC)C=C1N2 RERZNCLIYCABFS-UHFFFAOYSA-N 0.000 claims abstract description 23
- VJHLDRVYTQNASM-UHFFFAOYSA-N harmine Natural products CC1=CN=CC=2NC3=CC(=CC=C3C=21)OC VJHLDRVYTQNASM-UHFFFAOYSA-N 0.000 claims abstract description 23
- 239000002840 nitric oxide donor Substances 0.000 claims abstract description 13
- 238000012986 modification Methods 0.000 claims abstract description 7
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- 125000001917 2,4-dinitrophenyl group Chemical group [H]C1=C([H])C(=C([H])C(=C1*)[N+]([O-])=O)[N+]([O-])=O 0.000 description 1
- NEAQRZUHTPSBBM-UHFFFAOYSA-N 2-hydroxy-3,3-dimethyl-7-nitro-4h-isoquinolin-1-one Chemical class C1=C([N+]([O-])=O)C=C2C(=O)N(O)C(C)(C)CC2=C1 NEAQRZUHTPSBBM-UHFFFAOYSA-N 0.000 description 1
- QIWYEOCQAIGINY-UHFFFAOYSA-N 3-[[4-(benzenesulfonyl)-5-oxido-1,2,5-oxadiazol-5-ium-3-yl]oxy]propan-1-ol Chemical compound OCCCOC1=NO[N+]([O-])=C1S(=O)(=O)C1=CC=CC=C1 QIWYEOCQAIGINY-UHFFFAOYSA-N 0.000 description 1
- WGYFINWERLNPHR-UHFFFAOYSA-N 3-nitroanisole Chemical compound COC1=CC=CC([N+]([O-])=O)=C1 WGYFINWERLNPHR-UHFFFAOYSA-N 0.000 description 1
- BHVOYWSKJSRXRV-UHFFFAOYSA-N 4-[[4-(benzenesulfonyl)-5-oxido-1,2,5-oxadiazol-5-ium-3-yl]oxy]butan-1-ol Chemical compound OCCCCOC1=NO[N+]([O-])=C1S(=O)(=O)C1=CC=CC=C1 BHVOYWSKJSRXRV-UHFFFAOYSA-N 0.000 description 1
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- CLZISMQKJZCZDN-UHFFFAOYSA-N [benzotriazol-1-yloxy(dimethylamino)methylidene]-dimethylazanium Chemical compound C1=CC=C2N(OC(N(C)C)=[N+](C)C)N=NC2=C1 CLZISMQKJZCZDN-UHFFFAOYSA-N 0.000 description 1
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- NLFBCYMMUAKCPC-KQQUZDAGSA-N ethyl (e)-3-[3-amino-2-cyano-1-[(e)-3-ethoxy-3-oxoprop-1-enyl]sulfanyl-3-oxoprop-1-enyl]sulfanylprop-2-enoate Chemical compound CCOC(=O)\C=C\SC(=C(C#N)C(N)=O)S\C=C\C(=O)OCC NLFBCYMMUAKCPC-KQQUZDAGSA-N 0.000 description 1
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- 125000004194 piperazin-1-yl group Chemical group [H]N1C([H])([H])C([H])([H])N(*)C([H])([H])C1([H])[H] 0.000 description 1
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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
Abstract
The invention discloses a harmine derivative, a preparation method and application thereof, belonging to the field of pharmaceutical chemistry. The invention combines the structural modification of Harmine (HM) with NO donor to form new NO donor-harmine derivative. The NO donor-dehydroharmine derivative can release a large amount of NO in tumor cells, and the antiproliferative activity in the tumor cells is selectively and obviously enhanced along with the increase of the NO release, so that the influence on normal cells is smaller. Compared with the bulk drug HM and the NO donor, the anticancer cell proliferation effect of the novel compound NO donor-harmine derivative is obviously enhanced.
Description
Technical Field
The invention relates to the field of pharmaceutical chemistry, in particular to a harmine derivative, a preparation method and application thereof.
Background
Harmine (HM) is also known as haline or Ha Erming, is a beta-carboline alkaloid, is originally separated from Peganum harmala l, contains various alkaloids, and has a main active ingredient of Harmine, and has a wide pharmacological effect and a therapeutic effect on cardiovascular systems, respiratory systems and central nervous systems. In addition, harmine has antibacterial, antitumor, analgesic, antiinflammatory, antibacterial, antiviral, and insect repellent effects. However, the strong neurotoxicity of harmine affects the clinical application of harmine.
A nitric oxide donor refers to a compound that can release a certain amount of nitric oxide by the action of an enzyme or non-enzyme. Nitric Oxide (NO) plays an important role in various physiological and pathological processes in the body, such as participation in tumorigenesis and metastasis, maintenance of dynamic balance of micro-and macrovessels, participation in nerve signaling, regulation of immune inflammation, and the like; in the aspect of anti-tumor, the NO promotes tumor angiogenesis at low concentration, and inhibits tumor cells through direct or indirect action at high concentration; NO combines with oxygen free radical to generate a series of active nitrogen, which damages the protein, nucleic acid and other components in tumor cells to play an indirect role. It has now been found that NO donors having antitumor activity include furazan nitroxides, nitrates, azoglycol enolates, oximes, guanidines, NO-metal complexes, sterone imines, hydroxylamines, N-hydroxyureas, S-nitrosothiols, wherein furazan nitroxides are an important class of NO donors, have thermal stability, are capable of generating high levels of nitric oxide, and a variety of promising derivatives based on nitric oxide release from furazan have been investigated as anticancer drug candidates. There has been NO report on the study of the combination of harmine derivatives and NO donors.
Disclosure of Invention
The invention aims to provide a harmine derivative, a preparation method and application thereof, widen the variety of the harmine derivative and improve the antitumor activity of the harmine derivative.
In order to achieve the above object, the present invention provides the following solutions:
according to one technical scheme of the invention, the structural general formula of the harmine derivative is shown as formula I:
R 1 is NHC (CH) 3 )COOCH 3 、NHC(CH 3 ) 2 COOCH 3 Or O (CH) 2 ) n ON 2 SO 2 Ph, where n=3 or 4.
In a second technical scheme of the invention, the preparation method of the harmine derivative is that R 1 Is NHC (CH) 3 )COOCH 3 Or NHC (CH) 3 ) 2 COOCH 3 When the preparation method comprises the following stepsThe steps are as follows: carrying out structural modification on harmine to obtain the harmine derivative;
when R is 1 Is O (CH) 2 ) n ON 2 SO 2 At the time of Ph, the preparation method comprises the following steps: and carrying out structural modification on the harmine to obtain an intermediate product, and combining the intermediate product with a nitric oxide donor to obtain the harmine derivative.
Further, the structural formula of the intermediate product is shown as a formula II:
further, the NO donor includes furazan nitroxides, azodiol alkenylonium salts, organic nitrates, metal-NO complexes, stethones, N-hydroxyureas, and hydroxamic acids.
Further, the nitric oxide donor is furazan nitric oxide.
In the third technical scheme of the invention, the harmine derivative is applied to the preparation of antitumor drugs.
Further, when R 1 Is NHC (CH) 3 )COOCH 3 In the case of the anti-tumor drug, the anti-tumor drug is a drug for resisting human breast cancer MCF-7.
Further, when R 1 Is NHC (CH) 3 ) 2 COOCH 3 In the case, the antitumor drug is an anti-human gastric cancer cell BGC and/or anti-human breast cancer MCF-7 drug.
Further, when R 1 Is O (CH) 2 ) n ON 2 SO 2 In the Ph process, the antitumor drug is an antitumor drug of human liver cancer cell HepG2, human gastric cancer cell BGC, human breast cancer MCF-7 and/or human lung cancer cell A549.
The invention discloses the following technical effects:
(1) The invention takes harmine as raw material, and forms new harmine derivatives by carrying out structural modification and combining with NO donorTo improve the anti-tumor effect without neurotoxicity. All the structures are warp 1 H-NMR and MS confirmation. And a series of synthesized harmine derivatives are subjected to preliminary in vitro antitumor activity measurement by adopting a CCK-8 method.
(2) The invention synthesizes a new harmine derivative III for the first time 6a 、Ⅲ 6b 、Ⅳ 3a And IV 3b The harmine derivative III is verified by cell experiments 6a For tumor cells MCF-7, III 6b Has certain antiproliferative effect on tumor cells BGC, A549 and MCF-7; harmine derivatives IV 3a And IV 3b Has certain antiproliferative effect on tumor cells HepG2, BGC, A549 and MCF-7, wherein IV 3b The anti-tumor activity against HepG2 was best (IC 50 ＝1.79±0.16μM)。
(3) The invention widens the species of the harmine derivative, and provides a feasible way for improving the antitumor effect of the harmine derivative or the NO donor, and the harmine derivative with the modified structure is combined with the NO donor to form a novel NO donor-harmine derivative, so that a large amount of NO is released in tumor cells, and the high-concentration NO and the harmine derivative can play a synergistic antitumor effect, thereby improving the antitumor effect of the harmine in bulk drugs to a great extent. The harmine derivative prepared by the invention has NO obvious NO release in normal cells and has less influence on the normal cells.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are needed in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and other drawings can be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 shows the NO donor-harmine derivative IV prepared in example 1 3b Releasing NO in the HepG2 and LO2 liver cells;
FIG. 2 is a sample of NO donor-harmine derivative IV prepared in example 1 3b Under the condition of different concentrations, releasing NO in the HepG2 and LO2 cells of the human liver cancer cells;
FIG. 3 shows the NO donor-harmine derivative IV prepared in example 1 3b And releasing NO in the HepG2 and LO2 cells of the human liver cancer cells at different reaction times.
Detailed Description
Various exemplary embodiments of the invention will now be described in detail, which should not be considered as limiting the invention, but rather as more detailed descriptions of certain aspects, features and embodiments of the invention.
It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. In addition, for numerical ranges in this disclosure, it is understood that each intermediate value between the upper and lower limits of the ranges is also specifically disclosed. Every smaller range between any stated value or stated range, and any other stated value or intermediate value within the stated range, is also encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.
Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control.
It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the invention described herein without departing from the scope or spirit of the invention. Other embodiments will be apparent to those skilled in the art from consideration of the specification of the present invention. The specification and examples of the present invention are exemplary only.
As used herein, the terms "comprising," "including," "having," "containing," and the like are intended to be inclusive and mean an inclusion, but not limited to.
Example 1
NO donor-harmine derivative IV 3a 、Ⅳ 3b The specific synthetic route is shown below:
step 1, adding harmine HM (0.3 g,1.41 mmol) and 9mL dimethylformamide DMF into a 50mL three-necked flask, stirring until the mixture is clear under the protection of nitrogen, adding 60% NaH (0.07 g,2.82 mmol) and stirring for 0.5h under the ice bath state, dissolving 1-bromo-3-phenylpropane (2.12 mmol) in a small amount of DMF (diluting 1-bromo-3-phenylpropane to avoid too high concentration during the addition), reacting for 0.5h under the ice bath condition, removing the ice bath, stirring for 2h at room temperature, performing TLC tracking detection, extracting the reaction solution with ethyl acetate and water, concentrating under reduced pressure, purifying by silica gel column chromatography to obtain an off-white solid, namely an intermediate product II 1 。
Step 2, taking an intermediate product II 1 (0.3 g,0.91 mmol) is put into a 50mL round bottom flask, 7mL glacial acetic acid is added firstly for dissolution, then 7mL HBr is added for uniform mixing, the mixture is heated to 120 ℃ for reflux reaction for 12h, TLC tracking detection is carried out, after the reaction is finished, the reaction solution is poured into ice water, 10M sodium hydroxide aqueous solution is used for regulating pH to 6.0 under stirring, then sodium bicarbonate is used for regulating pH to 9.0, suction filtration, a large amount of water washing and silica gel column chromatography purification are carried out, and light yellow solid, namely an intermediate product II, is obtained 2 。
Step 3, intermediate product II 2 (0.05 g,0.16 mmol) was dissolved in DMF and then CS was added 2 CO 3 (0.07 g,0.24 mmol) was stirred at room temperature for 30min, and ethyl bromoacetate (0.05 mg,0.32 mmol) was added thereto to react at room temperature for 2 minh. TLC tracking detection, after the reaction is completed, extracting the reaction solution with ethyl acetate and water, concentrating under reduced pressure, purifying by silica gel column chromatography to obtain a tan solid, namely an intermediate product II 3 。
Step 4, taking an intermediate product II 3 (0.5 g,1.24 mmol) is dissolved by EA, benzyl bromide (2.1 g,12.4 mmol) is added, reflux is carried out for 12h at 90 ℃, TLC detection reaction is carried out, after the raw materials are reacted completely, cooling precipitation is carried out, suction filtration is carried out, ethyl acetate EA is used for washing for multiple times, thus obtaining white solid, and intermediate product II is obtained after drying 4 The next step was directly carried out without purification.
Step 5, taking intermediate product II 4 (0.3 g,0.6 mmol) lithium hydroxide (0.07 g,3.0 mmol), methanol 3mL, water 6mL, tetrahydrofuran (THF) 9mL were added and stirred at room temperature for 2h. TLC tracking detection, adding dilute hydrochloric acid to regulate pH to 4-5 after reaction, spin-drying on rotary evaporator to obtain intermediate product II 5 。
Step 6, taking an intermediate product II 5 (0.2 g,0.58 mmol) was dissolved in 5-6mL DMF and Et was added 3 After addition of N, TBTU (triethylamine; O-benzotriazol-N, N, N ', N' -tetramethyluronium tetrafluoroborate) (0.12 g,0.58 mmol) IV 2 The compound shown is (4- (4-hydroxybutoxy) -3- (benzenesulfonyl) -1,2, 5-oxadiazole 2-oxide, labeled iv when n=3 2a 4- (3-hydroxypropoxy) -3- (benzenesulfonyl) -1,2, 5-oxadiazole 2-oxide, labeled iv when n=4 2b ) Reacting for 3h at room temperature, purifying by column to obtain IV 3 。
When IV 2 When n in (3), the NO donor-harmine derivative IV obtained by the reaction 3 Marked as IV 3a : off-white solid, 100mg, yield 70%. 1 H NMR(600MHz,DMSO-d 6 )δ8.81(d,J＝6.5Hz,1H),8.62(d,J＝6.5Hz,1H),8.39(d,J＝8.8Hz,1H),8.01-7.97(m,2H),7.90-7.86(m,1H),7.75-7.70(m,2H),7.44-7.38(m,4H),7.23(t,J＝7.4Hz,2H),7.19-7.13(m,5H),6.04(s,2H),5.12(s,2H),4.67(d,J＝11.1Hz,2H),4.37(t,J＝6.1Hz,2H),4.30(t,J＝6.2Hz,2H),3.52(s,1H),2.95(s,2H),2.68(t,J＝7.8Hz,2H),1.15(s,3H)。
When IV 2 When n in (2) is 4, the reaction is carried outNO donor-harmine derivative IV 3 Marked as IV 3b : yellow-white solid, 80mg, yield 60%. 1 H NMR(600MHz,DMSO- d6 )δ8.80(s,1H),8.63(s,1H),8.42(s,1H),7.99(s,2H),7.88(d,J＝1.5Hz,2H),7.81-7.64(m,2H),7.50-7.31(m,4H),7.31-7.06(m,8H),6.03(s,2H),5.11(s,2H),4.80-4.61(m,2H),4.38(d,J＝6.1Hz,2H),4.24(t,J＝6.3Hz,2H),3.35(s,2H),2.96(s,3H),2.75-2.62(m,2H),2.50(s,2H),1.89-1.65(m,4H)。
The specific structural formula is as follows:
example 2
Harmine derivatives III 6a And III 6b The specific synthetic route is shown below:
the synthesis of intermediate products II 1, II 2, II 3 in steps 1,2, 3 was carried out in the same manner as in example 1.
Step 4, intermediate product II 3 (0.3 g,0.72 mmol) was mixed with lithium hydroxide (0.09 g,3.6 mmol), methanol 6mL, water 12mL and THF18 mL and stirred at room temperature for 2h. After the reaction shown by TLC is completed, dilute hydrochloric acid is added to adjust the pH value to 4-5, and the mixture is dried by spin on a rotary evaporator to obtain an intermediate product III 4 。
Step 5, taking intermediate product III 4 (0.05 g,0.14 mmol) was dissolved in 3-4mL of DMF, 2- (7-azabenzotriazol) -N, N, N ', N' -tetramethyluronium Hexafluorophosphate (HATU) (0.1 g,0.27 mmol), N, N-Diisopropylethylamine (DIPEA) (0.04 g,0.3 mmol) was added thereto, stirring at room temperature for 10min, then L-amino hydrochloride (0.16 mmol) was added thereto, the reaction was continued at room temperature for 30min, TLC was followed by detection, after completion of the reaction, the reaction solution was extracted with ethyl acetate and water, and concentrated under reduced pressure to give an off-white solid, namely intermediate III 5 . Purification by column chromatography to give intermediate III 5a And III 5b . The specific structural formula is as follows:
step 6, respectively taking the compound III 5a And III 5b Dissolving (0.3 g,0.54 mmol) with ethyl acetate, adding benzyl bromide (1.0 g,5.4 mmol) respectively, refluxing at 90deg.C for 12 hr, TLC detecting reaction, cooling to precipitate, vacuum filtering, washing with ethyl acetate for multiple times, and drying to obtain white solid harmine derivative III 6a And a pale yellow solid harmine derivative III 6b The specific structural formula is as follows:
harmine derivatives III 6a : off-white solid, 100mg, yield 95%. 1 H NMR(300MHz,DMSO-d6)δ8.80(d,J＝6.6Hz,1H),8.67(d,J＝12.1Hz,2H),8.43(d,J＝8.8Hz,1H),7.40(t,J＝7.3Hz,4H),7.19(dt,J＝13.2,6.9Hz,9H),6.03(s,2H),4.81(s,2H),4.74-4.61(m,2H),4.48-4.34(m,1H),3.63(s,3H),2.95(s,3H),2.69(t,J＝7.6Hz,2H),2.15–2.02(m,2H),1.36(d,J＝7.3Hz,2H)。
Harmine derivatives III 6b : pale yellow solid, 96mg, yield 93%. 1 HNMR(300MHz,DMSO-d 6 )δ8.79(d,J＝1.9Hz,1H),8.63(d,J＝6.5Hz,1H),8.45(dd,J＝17.9,9.2Hz,2H),7.47-7.29(m,4H),7.17(q,J＝7.5,6.9Hz,8H),6.01(s,2H),4.88(s,2H),4.75-4.59(m,2H),3.64(s,3H),2.94(s,3H),2.68(t,J＝7.6Hz,2H),2.13-2.04(m,2H),0.96-0.80(m,6H)。
Experimental example 1
Verification of harmine derivatives II 4 ，Ⅱ 5 ，Ⅲ 6a ，Ⅲ 6b ，Ⅳ 3a And IV 3b Proliferation inhibition activity on human tumor cell HepG2 (human liver cancer cell), BGC (human gastric cancer cell), A549 (human lung cancer cell), MCF-7 (human breast cancer cell), and selectingHM and Doxorubicin (DOX) were used as controls.
The method comprises the following steps:
four tumor cells were placed at 37℃and 5% CO 2 Culturing under the condition, digesting the grown cells with pancreatin containing EDTA, placing into a 5ml centrifuge tube, and centrifuging at 1200rpm for 5min. After centrifugation, the supernatant was removed, and diluted and mixed with 2ml of the culture medium. The cells in logarithmic growth phase were grown at 1X 10 5 cells/well were seeded in 96-well plates with PBS at four sides, 100. Mu.L of each well was added, and plating was completed. And (5) placing the mixture into a incubator for incubation for 24 hours. Each harmine derivative is subjected to gradient dilution, and the method comprises the following steps: HM, II 4, II 5 was diluted at an initial concentration of 200. Mu.M from a 2-fold gradient at high concentration to 6.25. Mu.M, i.e., 200. Mu.M, 100. Mu.M, 50. Mu.M, 25. Mu.M, 12.5. Mu.M, 6.25. Mu.M for a total of 6 concentration gradients; the same method as III 6a, III 6b, IV 3a, IV 3b and DOX was diluted in a 3-fold gradient from high concentration to 0.21. Mu.M at an initial concentration of 50. Mu.M, i.e., 50. Mu.M, 16.67. Mu.M, 5.56. Mu.M, 1.85. Mu.M, 0.62. Mu.M, 0.21. Mu.M for a total of 6 concentrations. . Taking out the paved 96-well plate from the incubator after 24 hours, sucking the culture medium out, adding the diluted sample into the 96-well plate with each concentration of 3 holes, respectively adding 100 mu L of the diluted sample into the 96-well plate with each hole, and putting the 96-well plate into a incubator with 37 ℃ and 5% CO 2 Is incubated for 72h. Then, the culture medium of the 96-well plate was aspirated off, and CCK-8 reagent (2- (2-methoxy-4-nitrobenzene) -3- (4-nitrobenzene) -5- (2, 4-disulfophenyl) -2H-tetrazolium monosodium salt) was added, followed by further culturing in an incubator for 30min, and detection was performed. Cell viability was calculated, cell viability (%) = (a) Sample of -A Blank space )/(A Negative of -A Blank space ) OD was measured at 450nm using a microplate reader. Results are expressed as mean+ -SD, IC was derived from inhibition by GraphPad Prism 6 fitting 50 . The results are shown in Table 1.
TABLE 1
As can be seen from Table 1, intermediate product II 4 、Ⅱ 5 IC for tumor cell HepG2 50 All are larger, explaining itThe antiproliferative activity against these two tumor cells is weak. While II 4 、Ⅱ 5 、Ⅲ 6a Exhibits stronger antiproliferative activity against BGC and MCF than HepG 2. III 6b Has better antiproliferative activity on tumor cells BGC and MCF-7 (IC) 50 ＝3.83±0.60μM；IC 50 =4.48±0.77 μm). New NO donor-harmine derivative IV formed by combining harmine derivative (intermediate product) with NO donor 3a And IV 3b Also exhibits potent antiproliferative activity on human cancer cells, wherein IV 3b The strongest antitumor activity (IC) 50 ＝1.79±0.16μM)。
Experimental example 2
Testing intermediate product II 5 Raw material drug IV of NO donor 2b NO donor-harmine derivative IV 3b And JSK (O) 2 - (2, 4-dinitrophenyl) 1- [ (4-ethoxycarbonyl) piperazin-1-yl]Azo-1-onium-1, 2-diol) releases NO in human hepatoma cell HepG2 and human normal hepatoma cell LO2 cells, specifically as follows: hepG2 and LO2 cells in logarithmic growth phase were 1X 10, respectively 7 cells/well were seeded in 96 well cell culture plates with PBS at four sides, 100. Mu.L of each well was added, plating was completed, and the cells/wells were incubated at 37℃with 5% CO 2 Incubator for 24h. After the incubation, the medium was removed with a pipette and the medium was removed at a concentration of 5. Mu.M II 5 、Ⅳ 2b 、Ⅳ 3b And JSK were added to LO2 and HepG2 cell culture plates, respectively, and continued at 37℃with 5% CO 2 Incubator incubates for 72h. After the incubation time was completed, the cell culture plate supernatant was removed, and after collecting cells, 1.5ml of 5. Mu.M DAF-FM DA (diaminofluorescein-FM diacetate, NO fluorescent probe) was added for 20min, and then washed 3 times with PBS buffer to remove excess dye. The intracellular NO level was determined by measuring the fluorescence intensity using a flow cytometer at an excitation/emission frequency of 495/515 nm. The results are shown in FIG. 1.
As can be seen from FIG. 1, intermediate product II is compared 5 NO donor drug substance IV 2b And a positive control drug JS-K, NO donor-harmine derivative IV 3b The NO level in the human hepatoma cell HepG2 is highest, the effect is strongest, and the human liver is normalThe difference in the amount of NO released by the above substances in the cells was not obvious, which indicates that NO donor-harmaline derivative IV 3b Has the strongest inhibition effect on human liver cancer cells and smaller influence on normal liver cells, and has better effect than the positive medicament JS-K.
Experimental example 3
Testing NO donor-harmine derivative IV at different concentrations 3b The conditions of releasing NO in the human liver cancer cell HepG2 and the human normal liver cell LO2 are as follows: hepG2 and LO2 cells in logarithmic growth phase were 1X 10, respectively 7 Inoculating cells/well into 96-well cell culture plate, adding 100 μl of sample into each well, plating, and placing into 37 deg.C and 5% CO 2 Incubator incubates for 24h. After the incubation, the medium was removed by pipetting gun and IV at 1,3, 9. Mu.M concentration, respectively 3b Respectively adding into LO2 and HepG2 cell culture plates, and continuing at 37deg.C, 5% CO 2 Incubator incubates for 72h. After the incubation time was completed, the cell culture plate supernatant was removed, and after collecting the cells, 1.5ml of 5. Mu.M DAF-FM DA was added for 20min, and washed 3 times with PBS buffer to remove excess dye. The intracellular NO levels were determined using flow cytometry at excitation/emission frequencies of 495/515nm to determine fluorescence intensity. The results are shown in FIG. 2.
As can be seen from fig. 2, the release of NO is dose dependent for the different dose groups, with the high dose group acting most significantly. Whereas different concentrations of NO donor-harmine derivative IV 3b The effect on human normal hepatocytes LO2 was not apparent in HepG2 cells, indicating that NO donor-harmine derivative IV 3b The effect of releasing NO in the human liver cancer cell HepG2 is strong and the influence on normal liver cells is small.
Experimental example 4
Testing NO donor-harmine derivative IV 3b Under different reaction time, the conditions of releasing NO in the HepG2 cell of the human liver cancer cell and the LO2 cell of the human normal liver cell are as follows: hepG2 and LO2 cells in logarithmic growth phase were 1X 10, respectively 7 cells/well were seeded in 96 well cell culture plates with PBS at four sides, 100. Mu.L of each well was added, plating was completed, and the cells/wells were incubated at 37℃with 5% CO 2 Incubator incubates for 24h. Incubation knotAfter the beam, the medium was removed with a pipette and IV was used at a concentration of 1. Mu.M 3b Added to LO2 and HepG2 cell culture plates, continued at 37℃with 5% CO 2 Incubating for 0h, 8h, 16h, 24h, 48h and 72h in incubator respectively. After the incubation time was completed, the cell culture plate supernatant was removed, and after collecting cells, 1.5ml of 5. Mu.M DAF-FM DA was added for 20min, and then washed 3 times with PBS buffer to remove excess dye. The fluorescence intensity was measured by flow cytometry at an excitation/emission frequency of 495/515nm, and the intracellular NO level was measured, and the results are shown in FIG. 3.
As can be seen from FIG. 3, NO donor-harmine derivative IV 3b The levels of NO release in HepG2 cells at 0h, 8h, 16h, 24h, 48h and 72h are time dependent, with significant differences starting from 16 h. Wherein the effect peaks at 72h. Whereas at different times the NO donor-harmine derivative IV 3b The effect on the normal liver cell LO2 of the human is not time dependent.
Test example 5
Acute toxicity evaluation
Kunming mice (available from Beijing Hua Biotechnology Co., ltd., license number: SCXK (Beijing) 2019-0008) were 18-22g in weight. The solvent used is physiological saline, 5% dimethyl sulfoxide (DMSO) and 5% Tween 80. After the sample is weighed, tween 80 is added to assist dissolution, and physiological saline containing 5% DMSO is added to the required concentration. Mice were randomly divided into 3 groups of 5 mice each. HM and IV 3b doses are 150.0mg/kg,50.0mg/kg,10.0mg/kg for the high, medium and low groups, respectively; the tail vein was administered 1 time, and the administration was followed by continuous observation for 7 days. The general status, activity, feeding of mice were observed to record the death status, toxic symptoms and start-stop time of toxic response of mice. Mice developed toxic response times and death times were slightly different after each subject was dosed. The results are shown in Table 2.
TABLE 2
Mice in the HM high dose (150 mg/kg) group exhibited convulsions, keratotic archesReverse tension, body stiffness, total death within 0-5 mm; administration IV 3b Mice in the high dose (150 mg/kg) group showed convulsions and dyspnea, gradually dying completely within 40 min. Two mice at the dose (50 mg/kg) in HM die within 30min and return to normal after the remaining 30 min. Administration IV 3b Medium dose (50 mg/kg) and low dose (10 mg/kg) mice exhibited reduced tremor excitability, but did not die within 7d. Compared with the bulk drug HM, the novel compound NO donor-harmine derivative obviously reduces the acute toxicity of mice.
The invention verifies the novel compound harmine derivative III through the cellular level 6a 、Ⅲ 6b 、Ⅱ 4 And II 5 New compound NO donor-harmine derivative IV 3a And IV 3b The antiproliferative effect of the novel compounds on HepG2, BGC, A549 and MCF-7 is improved to different degrees on the antiproliferative activity of the novel compounds on the four cells compared with the HM of the crude drugs according to experimental results, and meanwhile, compared with the HM, the antiproliferative effect of the novel compounds on the LO2 of normal cells is weakened, namely the toxicity on the LO2 is reduced to a certain degree. Wherein the novel compound NO donor-harmine derivative IV 3b The antiproliferative effect on HepG2 was strongest, so we further examined at the cellular level in order to explore the relationship of NO to antiproliferative activity. As shown in FIG. 1, the drug JS-K, II is compared with the positive control drug 5 ，Ⅳ 2b Compound IV with strongest antiproliferative activity compared to the blank group 3b The relative MFI was highest in the hepatoma cell line HepG 2. Thus, the amount of NO released in HepG2 cells correlates with antiproliferative activity. Under the same conditions, compound IV 3b (1.5. Mu.M, 72 h) significantly increased the relative Mean Fluorescence Intensity (MFI) of cancer cells HepG2 but not LO2 cells, indicating that NO was selectively released only in human hepatoma cells HepG2 and normal human hepatoma cells LO2 were hardly affected. As shown in FIG. 2, as the HepG2 cell concentration increases, compound IV 3b The released NO showed a more pronounced concentration dependence than LO2 cells. Also, as shown in FIG. 3, in the presence of compound IV 3b After 72h incubation, NO release was maximized in HepG2 cells, whereas NO change was not evident in LO2 cells. Above mentionedDescription of Compounds IV 3b Has certain selectivity in releasing NO to exert the antiproliferative effect, namely has obvious antiproliferative effect in HepG2 cells, and has smaller influence on LO2 of normal cells.
The above embodiments are only illustrative of the preferred embodiments of the present invention and are not intended to limit the scope of the present invention, and various modifications and improvements made by those skilled in the art to the technical solutions of the present invention should fall within the protection scope defined by the claims of the present invention without departing from the design spirit of the present invention.
Claims (4)
1. The harmine derivative is characterized by having a structural general formula as shown in formula I:
R 1 is thatWhere n=3 or 4.
2. The method for preparing a harmine derivative according to claim 1, characterized in that the method comprises the steps of: carrying out structural modification on harmine to obtain an intermediate product, and combining the intermediate product with a nitric oxide donor to obtain the harmine derivative;
the nitric oxide donor is
3. The method for preparing harmine derivatives according to claim 2, wherein the intermediate product has the structural formula shown in formula ii:
4. the use of the harmine derivative according to claim 1 for preparing an anti-tumor drug, wherein the anti-tumor drug is an anti-human liver cancer cell HepG2, an anti-human stomach cancer cell BGC, an anti-human breast cancer MCF-7 and/or an anti-human lung cancer cell a 549.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101429198A (en) * | 2007-11-09 | 2009-05-13 | 新疆华世丹药物研究有限责任公司 | Banisterine derivant and uses thereof |
| CN101775059A (en) * | 2010-02-04 | 2010-07-14 | 中国药科大学 | Novel glycyrrhetinic acid derivative, and preparation method and medicinal uses thereof |
| CN102977096A (en) * | 2012-12-07 | 2013-03-20 | 中国药科大学 | Harmine derivative anti-tumor prodrug with target characteristic |
| CN111072682A (en) * | 2019-11-07 | 2020-04-28 | 沈阳药科大学 | Chelidonine furazan nitric oxide donor derivative and preparation method and application thereof |
| CN111374975A (en) * | 2018-12-27 | 2020-07-07 | 北京大学 | Medicinal application of harmine |
-
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- 2021-12-20 CN CN202111560905.0A patent/CN114133390B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101429198A (en) * | 2007-11-09 | 2009-05-13 | 新疆华世丹药物研究有限责任公司 | Banisterine derivant and uses thereof |
| CN101775059A (en) * | 2010-02-04 | 2010-07-14 | 中国药科大学 | Novel glycyrrhetinic acid derivative, and preparation method and medicinal uses thereof |
| CN102977096A (en) * | 2012-12-07 | 2013-03-20 | 中国药科大学 | Harmine derivative anti-tumor prodrug with target characteristic |
| CN111374975A (en) * | 2018-12-27 | 2020-07-07 | 北京大学 | Medicinal application of harmine |
| CN111072682A (en) * | 2019-11-07 | 2020-04-28 | 沈阳药科大学 | Chelidonine furazan nitric oxide donor derivative and preparation method and application thereof |
Non-Patent Citations (3)
| Title |
|---|
| "Nitric oxide donor hybrid compounds as promising anticancer agents";Ding Q G 等;《Drug Discoveries & Therapeutics》;第10卷(第6期);第276-284页 * |
| "Pharmacological effects of harmine and its derivatives: a review";Zhang L等;《Archives of Pharmacal Research》;第43卷(第12期);第1259-1275页 * |
| "Synthesis and biological evaluation of nitric oxide-releasing matrine derivatives as anticancer agents";Li Qin He 等;《Chinese Chemical Letters》;第21卷(第4期);第381-384页 * |
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