CN105693729B

CN105693729B - Indoles simultaneously [3,2 a] carbazole derivates and its application

Info

Publication number: CN105693729B
Application number: CN201510893281.2A
Authority: CN
Inventors: 刘晟; 本大卫·雅科夫; 肖潇; 徐梅; 杨超; 梁丽娜; 范田运; 韩飞; 刘唐婧君; 郝小江
Original assignee: Key Laboratory of Natural Product Chemistry of Guizhou Academy of Sciences
Current assignee: Key Laboratory of Natural Product Chemistry of Guizhou Academy of Sciences
Priority date: 2015-12-08
Filing date: 2015-12-08
Publication date: 2017-12-22
Anticipated expiration: 2035-12-08
Also published as: CN105693729A

Abstract

The present invention relates to medicinal chemistry art, specially discloses a kind of such as formulaShown indoles simultaneously [3,22 a] carbazole derivates are as the application for preparing antineoplastic, each substituent is defined in the specification in formula, to human erythroleukemia cell HEL, leukaemia K 562 1, human melanoma cell WM 91, breast cancer cell MDA MB 231, human lymphoblastoid TK 6, prostate gland cancer cell PC 3 are respectively provided with good activity, have preferable Prospect of R ＆ D.

Description

Indoles simultaneously [3,2-a] carbazole derivates and its application

Technical field

The present invention relates to chemical medicine field, specifically using tryptophan methyl ester as starting material, synthesizes a series of Yin Diindyl simultaneously [3,2-a] carbazole derivates and derivative salt and its application in antineoplastic is prepared.

Background technology

Tumour always is the big threat of human life, and the struggle of the mankind and tumour is continuing always, wherein the most Effective method is exactly drug therapy, and the phenomenon of poorly efficient high poison in many medicines for the treatment of tumour be present, thus exploitation and Research is always an important directions of present field of medicaments with antitumor activity and the relatively low medicine of toxicity, association area Researchers be directed to finding the compound of a kind of effectively low toxicity always, this great problem of the mankind is addressed with it.

Simultaneously [3,2-a] carbazole alkaloid is more novel natural products to indoles, until ability in 2002 from It is found in marine organisms sponge, there is extensive bioactivity particularly antitumor activity, enjoy relatively broad pass always Note, but because synthetic method is less and natural origin is limited, the medicine and activity research so far around this compound are seldom. The present inventor has developed a synthetic method for such carbazole alkaloid in the recent period, can conveniently obtain different groups Substituted indoles simultaneously [3,2-a] carbazole derivates.In research before, to indoles simultaneously [3,2-a] The activity against organisms research of carbazole alkaloid is seldom, especially relatively fewer to the report of its antitumor activity, to antitumor Species it is also relatively fewer, in order to expand the chemical space of such compound and biology space, utilize indoles simultaneously [3,2 - a] as molecular template, one is designed and synthesizes by introducing pharmacophore carbazole this more novel natural products parent nucleus The compound of series, it is desirable to which on the basis of keeping activity and reducing toxicity, synthesis has preferable physicochemical property and metabolic stability Property medicine as lead compound, and study its bioactivity to kinds of tumor cells, be the exploitation of anti-cancer agent Material base is provided.

The content of the invention

The purpose of the present invention is to utilize indoles simultaneously this more novel natural products parent nucleus of [3,2-a] carbazole As molecular template, a series of compound is designed and synthesized by introducing pharmacophore, prepares new, various structures indoles And [3,2-a] carbazole derivates and its pharmaceutically acceptable salt, research, which has, resists various tumor promotions, is swollen to be anti- The preparation of knurl new drug provides material base.

A kind of indoles of the present invention simultaneously [3,2-a] carbazole derivates, it is characterised in that its general structure is formula （1）：

Formula（1）In：

R₁For hydrogen, hydroxyl,；

R₂For hydrogen, halogen atom, hydroxyl, the alkoxies of 1~C of C 4, the alkanoyls of 1~C of C 4；

R₃For by formula（ 2 ）Shown substituted radical：

Formula（ 2 ）In, n=1~6, R is selected from alkylamino, amide groups, Heterocyclylalkyl, aryl, substituted-phenyl, virtue Heterocyclic radical；Simultaneously [3,2-a] carbazole derivates abbreviation derivative also includes its salt generated to signified indoles；Signified Derivative or derivative salt have the activity for suppressing tumour.

Simultaneously 4. [3,2-a] carbazole derivates derivative salt is that can pharmaceutically connect to a kind of indoles indicated above Hydrochloride, sulfate, mesylate, tosilate, maleate, fumarate, the tartrate received；Specifically Above-mentioned formula（1）Middle R₁Represent R₁Represent hydrogen, hydroxyl,；

R₂Represent hydrogen, methoxyl group, chlorine or bromine；R₃Represent, And derivative salt is pharmaceutically acceptable salt hydrochlorate, derivative or derivative salt have antitumor activity；The part of synthesis Compound or its salt such as table 1, wherein it is preferred that the compound C 1 synthesized, C 3, C 11, C 12, C 13, C 14, C 15, C 16, C 27, C 28 or its hydrochloride have significant antitumor activity；This kind of indoles simultaneously [3,2-a] carbazole The application of derivative, it is to be used to prepare anti-human erythroleukemia cell HEL, myelogenous leukemia cells K 562-1, human melanoma Cell WM9-1, breast cancer cell MDA 231, B lymphoblasts TK-6, FV-P induction erythroleukemia (mouse cell) The medicine or medicament of these plastidogenetic tumours of DP 17-17, prostate gland cancer cell PC-3.

Table 1：The part of compounds of synthesis

The nuclear magnetic data of the compound of 2 table of table 1 characterizes

It is preferred that C 1, C 3, C 11, C 12, C 13, C 14, C 15, C 16, C 27, C 28, excellent to this ten kinds Simultaneously [3,2-a] carbazole derivates will carry out embodiment explanation to the indoles of choosing.

The antitumor experiment of in vitro and in vivo is carried out to compound, shows that it has the significant activity for suppressing tumour, because This compounds of this invention can be used for preparing anti-tumor drug or medicament.

Invention effect：In the compound that the present invention has synthesized, preferably go out C 1, C 3, C 11, C 12, C 13, C 14th, C 15, C 16, C 27, C 28 or its salt are representative, to human erythroleukemia cell HEL, myelogenous leukemia cells K 562-1, human melanoma cell WM9-1, breast cancer cell MDA 231, B lymphoblasts TK-6, FV-P induce red Leukaemia (mouse cell) DP 17-17, prostate gland cancer cell PC-3 kinds of tumor cells carry out drug study, find equal With significant antitumor action.

Figure of description

Fig. 1 is raw material F synthetic route；

Fig. 2 is raw material M synthetic route；

Fig. 3 is raw material P synthetic route；

Fig. 4 is raw material U synthetic route；

Fig. 5 is influences of the C 1 to the cell growths of K 562 in embodiment 12；It can be seen that compound C 1 Concentration has more obvious inhibitory action with being proportionate to the inhibiting rate of the cell growths of K 562, to the cells of K 562；

Fig. 6 is influences of the C 1 in 72 h to the cell cycles of K 562 in embodiment 13；Therefrom can be with Find out that C 1 has obvious influence to the cell cycles of K 562, block in the G2 phases；

Fig. 7 is C 1 in embodiment 14（1 mg/kg) two weeks to infect F-MuLV mouse spleens The influence of weight；It can be seen that the spleen of ill mouse returns to normal, also just say compound C 1 to infection F- It is more substantially therapeutic action that MuLV mouse, which has,；

Fig. 8 is C 1 in embodiment 14（1 mg/kg) two weeks are red thin to infection F-MuLV mouse The influence of born of the same parents' specific volume, there it can be seen that the hematocrit value of the mouse of disease returns to normal, illustrate compound C 1 to sense It is more substantially therapeutic action that dye F-MuLV mouse, which has,.

Embodiment

The present invention is elaborated, it is necessary to which explanation is with reference to instantiation, following embodiments are only for It is bright, and it is not intended to limit the present invention.The various change that those skilled in the art are made according to the teachings of the present invention all should be Within protection domain required by the application claim.

Raw material F preparation

Fig. 1 is prepare compound F technology path.Specific method：Weigh Compound A（ 4.03 mmol ）It is molten In the super dry tetrahydrofurans of 10 mL, triethylamine is added（4.8 mmol, 1.2 eq）, then added at -78 DEG C secondary Chloric acid tertiary butyl ester（ 4.83 mmol, 1.2 eq ）, after reacting 40 min, sequentially add compound B（ 8.06 mmol, 2.0 eq ）And boron trifluoride^.Ether（ 16.09 mmol, 4.0 eq ）, 5 h of reaction are warmed to room temperature, are slowly added to saturation Sodium bicarbonate aqueous solution adjust pH=7, ethyl acetate extraction（ 3 × 10 mL ）, merge organic layer, anhydrous magnesium sulfate is done Dry, filtering, be concentrated under reduced pressure to obtain crude product, then obtains compound C through column chromatographic isolation and purification；By compound C（ 0.97 mmol ）It is dissolved in ethanol/methylene（ v / v = 1:1）In the mixed solvent, add 80% hydrazine hydrate（3.88 mmol, 4.0 eq）, react at room temperature overnight, filtering, take filtrate decompression to be concentrated to give crude product, then obtained through column chromatographic isolation and purification Compound D；Compound D（ 0.70 mmol ）It is dissolved in acetonitrile/acetate buffer solution（ 4 mL, v / v = 1:1）It is mixed In bonding solvent, Zinc vitriol is added（ 0.35 mmol ）And sodiam glyoxlate（ 7.0 mmol ）, 0.5 h is reacted at room temperature, Then add 10 mL water quenchings to go out reaction, ethyl acetate extraction（ 3 × 10 mL ）, merge organic layer, anhydrous Na₂SO₄It is dry It is dry, filter, be concentrated under reduced pressure to obtain crude product, this crude product is dissolved in 3mL Isosorbide-5-Nitrae-dioxane, then adds 0.1 ML trifluoroacetic acids, the h of heating reflux reaction 5-8, after cooling, are concentrated under reduced pressure, and mesh compound is obtained through column chromatographic isolation and purification E ；Weigh Compound E（ 0.38 mmol）It is dissolved in MeOH/H₂O / DMSO （ 2 mL , v / v / v = 2:1: 1 ）In the mixed solvent, add 3.8 mmol NaOH, react at room temperature 26-30 h（TLC monitoring reactions are complete）, Ran Houjia Enter the % of 3 mL 5 hydrochloric acid solution, ethyl acetate extraction, merge organic phase, anhydrous Na₂SO₄Dry, filtering, take filtrate decompression Enriched compound F.

Embodiment 1

C 1 preparation：

The technology path for preparing C 1 is：

Concrete operation method is：0.2 mmol compounds F is dissolved in dry DMF, adds 0.25 mmol HOBt·H₂O and 0.38 mmol EDCI, 0.5 mmol Et are sequentially added after 20 min are stirred at room temperature₃N and 0.25 Mmol compound G, are stirred overnight at room temperature.20 mL saturations Na are added into reaction system₂CO₃, ethyl acetate is extracted twice （ 2 × 10 mL ）, merge organic phase, washing, anhydrous Na₂SO₄Dry, filtering, take filtrate decompression to be concentrated to give crude product, then Purify to obtain the mg of white solid 59.3 through column chromatography, yield is 80 %;

C 1 NMR data are shown in Table 2.

Embodiment 2

C 3 preparation：

The technology path for preparing C 3 is：

Concrete operation method is：0.2 mmol compounds F is dissolved in dry DMF, adds 0.25 mmol HOBt·H₂O and 0.38 mmol EDCI, 0.5 mmol Et are sequentially added after 20 min are stirred at room temperature₃N and 0.25 Mmol compound H, are stirred overnight at room temperature.20 mL saturations Na are added into reaction system₂CO₃, ethyl acetate is extracted twice （ 2 × 10 mL）, merge organic phase, washing, anhydrous Na₂SO₄Dry, filtering, take filtrate decompression to be concentrated to give crude product, then Purify to obtain the mg of yellow solid 57.8, yield 73% through column chromatography;

C 3 NMR data are shown in Table 2.

Raw material M preparation

Fig. 2 is raw material M technology of preparing route concrete operation method：Weigh Compound I（ 4.03 mmol ）It is molten In the super dry tetrahydrofurans of 10 mL, triethylamine is added（4.8 mmol, 1.2 eq）, then added at -78 DEG C secondary Chloric acid tertiary butyl ester（ 4.83 mmol, 1.2 eq ）, after reacting 40 min, sequentially add compound B（ 8.06 mmol, 2.0 eq ）And boron trifluoride^.Ether（ 16.09 mmol, 4.0 eq ）, 5 h of reaction are warmed to room temperature, are slowly added to saturation Sodium bicarbonate aqueous solution adjust pH=7, ethyl acetate extraction（ 3 × 10 mL ）, merge organic layer, anhydrous magnesium sulfate is done Dry, filtering, be concentrated under reduced pressure to obtain crude product, then obtains compound J through column chromatographic isolation and purification；By compound J（ 0.97 mmol ）It is dissolved in ethanol/methylene（ v / v = 1:1）In the mixed solvent, add 80% hydrazine hydrate（3.88 mmol, 4.0 eq）, react at room temperature overnight, filtering, take filtrate decompression to be concentrated to give crude product, then obtained through column chromatographic isolation and purification Compound K；Compound K（ 0.70 mmol ）It is dissolved in acetonitrile/acetate buffer solution（ 4 mL, v / v = 1:1）It is mixed In bonding solvent, Zinc vitriol is added（ 0.35 mmol ）And sodiam glyoxlate（ 7.0 mmol ）, 0.5 h is reacted at room temperature, Then add 10 mL water quenchings to go out reaction, ethyl acetate extraction（ 3 × 10 mL ）, merge organic layer, anhydrous Na₂SO₄It is dry It is dry, filter, be concentrated under reduced pressure to obtain crude product, this crude product is dissolved in 3mL Isosorbide-5-Nitrae-dioxane, then adds 0.1 ML trifluoroacetic acids, the h of heating reflux reaction 5-8, after cooling, are concentrated under reduced pressure, and mesh compound is obtained through column chromatographic isolation and purification L；Weigh Compound L（ 0.38 mmol）It is dissolved in MeOH/H₂O / DMSO （ 2 mL , v / v / v = 2:1: 1 ）In the mixed solvent, add 3.8 mmol NaOH, react at room temperature 26-30 h（TLC monitoring reactions are complete）, Ran Houjia Enter the % of 3 mL 5 hydrochloric acid solution, ethyl acetate extraction, merge organic phase, anhydrous Na₂SO₄Dry, filtering, take filtrate decompression Enriched compound M.

Embodiment 3

C 11 preparation

The technology path for preparing C 11 is：

Concrete operation method is：0.2 mmol compounds M is dissolved in dry DMF, adds 0.25 mmol HOBt·H₂O and 0.38 mmol EDCI, 0.5 mmol Et are sequentially added after 20 min are stirred at room temperature₃N and 0.25 Mmol compound G, are stirred overnight at room temperature.20 mL saturations Na are added into reaction system₂CO₃, ethyl acetate is extracted twice （ 2 × 10 mL ）, merge organic phase, washing, anhydrous Na₂SO₄Dry, filtering, take filtrate decompression to be concentrated to give crude product, then Purify to obtain the mg of white solid 50.2, yield 65% through column chromatography;

C 11 NMR data are shown in Table 2.

Embodiment 4

C 12 preparation

The technology path for preparing C 12 is：

Concrete operation method is：0.2 mmol compounds M is dissolved in dry DMF, adds 0.25 mmol HOBt·H₂O and 0.38 mmol EDCI, 0.5 mmol Et are sequentially added after 20 min are stirred at room temperature₃N and 0.25 Mmol compound H, are stirred overnight at room temperature.20 mL saturations Na are added into reaction system₂CO₃, ethyl acetate is extracted twice （ 2 × 10 mL）, merge organic phase, washing, anhydrous Na₂SO₄Dry, filtering, take filtrate decompression to be concentrated to give crude product, then Purify to obtain the mg of yellow solid 58.6 through column chromatography, yield is 71 %;

C 12 NMR data are shown in Table 2.

Raw material P preparation

Fig. 3 is prepare compound P technology path.Concrete operation method：Weigh Compound I（ 4.03 mmol ） It is dissolved in the super dry tetrahydrofurans of 10 mL, adds triethylamine（4.8 mmol, 1.2 eq）, then added at -78 DEG C Hypochlorous acid tertiary butyl ester（ 4.83 mmol, 1.2 eq ）, after reacting 40 min, sequentially add compound B（ 8.06 mmol, 2.0 eq ）And boron trifluoride^.Ether（ 16.09 mmol, 4.0 eq ）, 5 h of reaction are warmed to room temperature, are slowly added The sodium bicarbonate aqueous solution for entering saturation adjusts pH=7, ethyl acetate extraction（ 3 × 10 mL ）, merge organic layer, anhydrous sulphur Sour magnesium is dried, and filtering, be concentrated under reduced pressure to obtain crude product, then obtains compound J through column chromatographic isolation and purification；By compound J（ 0.97 mmol ）It is dissolved in ethanol/methylene（ v / v = 1:1）In the mixed solvent, add 80% hydrazine hydrate（3.88 mmol, 4.0 eq）, react at room temperature overnight, filtering, take filtrate decompression to be concentrated to give crude product, then obtained through column chromatographic isolation and purification Compound K；Compound K（ 0.70 mmol ）It is dissolved in acetonitrile/acetate buffer solution（ 4 mL, v / v = 1:1）It is mixed In bonding solvent, Zinc vitriol is added（ 0.35 mmol ）And sodiam glyoxlate（ 7.0 mmol ）, 0.5 h is reacted at room temperature, Then add 10 mL water quenchings to go out reaction, ethyl acetate extraction（ 3 × 10 mL ）, merge organic layer, anhydrous Na₂SO₄It is dry It is dry, filter, be concentrated under reduced pressure to obtain crude product, this crude product is dissolved in 3mL Isosorbide-5-Nitrae-dioxane, then adds 0.1 ML trifluoroacetic acids, the h of heating reflux reaction 5-8, after cooling, are concentrated under reduced pressure, and mesh compound is obtained through column chromatographic isolation and purification L；Compound L and compound N are dissolved in 3 mL acetone, add potassium carbonate（4.0 eq ）, tetrabutylammonium iodide（1.2 eq ）, 5 h are heated to reflux, decompression boils off acetone, adds 10 mL water, ethyl acetate extraction（3 × 10 mL）, it is associated with Machine layer, anhydrous Na₂SO₄Dry, filtering, be concentrated under reduced pressure, compound O, Weigh Compound O are purified to obtain through column chromatography（ 0.38 mmol）It is dissolved in MeOH/H₂O / DMSO （ 2 mL , v / v / v = 2:1:1 ）In the mixed solvent, add 3.8 Mmol NaOH, react at room temperature 26-30 h（TLC monitoring reactions are complete）, then add the % of 3 mL 5 hydrochloric acid solution, second Acetoacetic ester extracts, and merges organic phase, anhydrous Na₂SO₄Dry, filtering, take filtrate decompression to be concentrated to give compound P.

Embodiment 5

C 13 preparation：

The technology path for preparing C 13 is：

Concrete operation method is：0.20 mmol compounds P is dissolved in dry DMF, adds 0.25 mmol HOBt·H₂O and 0.38 mmol EDCI, 0.5 mmol Et are sequentially added after 20 min are stirred at room temperature₃N and 0.25 Mmol compound G, are stirred overnight at room temperature.20 mL saturations Na are added into reaction system₂CO₃, ethyl acetate is extracted twice （ 2 × 10 mL）, merge organic phase, washing, anhydrous Na₂SO₄Dry, filtering, take filtrate decompression to be concentrated to give crude product, then Purify to obtain the mg of yellow solid 47.5, the % of yield 52 through column chromatography.

C 13 NMR data are shown in Table 2.

Embodiment 6

C 14 preparation：

The technology path for preparing C 14 is：

Concrete operation method is：0.20 mmol compounds P is dissolved in dry DMF, adds 0.25 mmol HOBt·H₂O and 0.38 mmol EDCI, 0.5 mmol Et are sequentially added after 20 min are stirred at room temperature₃N and 0.25 Mmol compound H, are stirred overnight at room temperature.20 mL saturations Na are added into reaction system₂CO₃, ethyl acetate is extracted twice （ 2 × 10 mL）, merge organic phase, washing, anhydrous Na₂SO₄Dry, filtering, take filtrate decompression to be concentrated to give crude product, then Purify to obtain the mg of yellow solid 39.6, the % of yield 41 through column chromatography.

C 14 NMR data are shown in Table 2.

Raw material U preparation

Fig. 4 is prepare compound U technology path.Concrete operation method：Weigh Compound A（ 4.03 mmol ） It is dissolved in the super dry tetrahydrofurans of 10 mL, adds triethylamine（4.8 mmol, 1.2 eq）, then added at -78 DEG C Hypochlorous acid tertiary butyl ester（ 4.83 mmol, 1.2 eq ）, after reacting 40 min, sequentially add compound Q（ 8.06 mmol, 2.0 eq ）And boron trifluoride^.Ether（ 16.09 mmol, 4.0 eq ）, 5 h of reaction are warmed to room temperature, are slowly added The sodium bicarbonate aqueous solution for entering saturation adjusts pH=7, ethyl acetate extraction（ 3 × 10 mL ）, merge organic layer, anhydrous sulphur Sour magnesium is dried, and filtering, be concentrated under reduced pressure to obtain crude product, then obtains compound R through column chromatographic isolation and purification；By compound R（ 0.97 mmol ）It is dissolved in ethanol/methylene（ v / v = 1:1）In the mixed solvent, add 80% hydrazine hydrate（3.88 mmol, 4.0 eq）, react at room temperature overnight, filtering, take filtrate decompression to be concentrated to give crude product, then obtained through column chromatographic isolation and purification Compound S；Compound S（ 0.70 mmol ）It is dissolved in acetonitrile/acetate buffer solution（ 4 mL, v / v = 1:1）Mixing In solvent, Zinc vitriol is added（ 0.35 mmol ）And sodiam glyoxlate（ 7.0 mmol ）, 0.5 h is reacted at room temperature, so After add 10 mL water quenchings and go out reaction, ethyl acetate extraction（ 3 × 10 mL ）, merge organic layer, anhydrous Na₂SO₄Dry, Filter, be concentrated under reduced pressure to obtain crude product, and this crude product is dissolved in 3mL Isosorbide-5-Nitrae-dioxane, then add 0.1 mL tri- Fluoroacetic acid, the h of heating reflux reaction 5-8, after cooling, is concentrated under reduced pressure, and mesh compound T is obtained through column chromatographic isolation and purification；Claim Take compound T（ 0.38 mmol）It is dissolved in MeOH/H₂O / DMSO （ 2 mL , v / v / v = 2:1:1 ）'s In the mixed solvent, 3.8 mmol NaOH are added, react at room temperature 26-30 h（TLC monitoring reactions are complete）, then add 3 The % of mL 5 hydrochloric acid solution, ethyl acetate extraction, merges organic phase, anhydrous Na₂SO₄Dry, filtering, take filtrate decompression to concentrate Obtain compound U.

Embodiment 7

C 15 preparation：

The technology path for preparing C 15 is：

Concrete operation method is：0.2 mmol compounds U is dissolved in dry DMF, adds 0.25 mmol HOBt·H₂O and 0.38 mmol EDCI, 0.5 mmol Et are sequentially added after 20 min are stirred at room temperature₃N and 0.25 Mmol compound G, are stirred overnight at room temperature.20 mL saturations Na are added into reaction system₂CO₃, ethyl acetate is extracted twice （ 2 × 10 mL ）, merge organic phase, washing, anhydrous Na₂SO₄Dry, filtering, take filtrate decompression to be concentrated to give crude product, then Purify to obtain the mg of white solid 64.9, yield 81% through column chromatography;

C 15 NMR data are shown in Table 2.

Embodiment 8

C 16 preparation：

The technology path for preparing C 16 is：

Concrete operation method is：0.2 mmol compounds U is dissolved in dry DMF, adds 0.25 mmol HOBt·H₂O and 0.38 mmol EDCI, 0.5 mmol Et are sequentially added after 20 min are stirred at room temperature₃N and 0.25 Mmol compound H, are stirred overnight at room temperature.20 mL saturations Na are added into reaction system₂CO₃, ethyl acetate is extracted twice （ 2 × 10 mL）, merge organic phase, washing, anhydrous Na₂SO₄Dry, filtering, take filtrate decompression to be concentrated to give crude product, then Purify to obtain the mg of yellow solid 72.5 through column chromatography, yield is 85 %;

C 16 NMR data are shown in Table 2.

Embodiment 9

C 27 preparation：

Concrete operation method is：Weigh 50 mg C 13（ 0.11 mmol ）It is dissolved in 2 mL CH₂Cl₂, add 0.5 mL The hydrochloric acid solution of dioxane, 30 min are stirred at room temperature, are spin-dried for solvent and just obtain the mg of yellow solid 54, the % of yield 100.

Embodiment 10

C 28 preparation：

Concrete operation method is：Weigh 50 mg C 14（0.10 mmol ）It is dissolved in 2 mL CH₂Cl₂, add 0.5 mL The hydrochloric acid solution of dioxane, 30 min are stirred at room temperature, are spin-dried for solvent and just obtain the mg of yellow solid 57.4, yield 100 %。

The beneficial effect of medicine of the present invention is expanded on further below by way of pharmacodynamics test and contrast test.

Tumor cell line the human erythroleukemia cell HEL, myelogenous leukemia cells K 562 that present invention experiment uses- 1, human melanoma cell WM9-1, breast cancer cell MDA 231, B lymphoblasts TK-6, FV-P induction erythroleukemia (mouse cell) DP 17-17, prostate gland cancer cell PC-3 provides for Shanghai cell bank.

Embodiment 11

Preferable C 1, C 3, C 11, C 12, C 13, C 14, C 15, C 16, C are determined using MTT methods 27th, the antitumor activity of this ten kinds of compounds of C 28.

Specific method：Take the logarithm people human erythroleukemia cell HEL, the myelogenous leukemia cells K 562-1 in growth period, Human melanoma cell WM9-1, breast cancer cell MDA 231, B lymphoblasts TK-6, FV-P induction erythroleukemia (mouse cell) DP 17-17, prostate gland cancer cell PC-3, it is inoculated with respectively with 2 × 104/mL cell densities In 96 well culture plates, 100 μ L/hole, every kind of various 4 blocks of plates of cell.37 DEG C are put, 5 % CO₂Trained in incubator Support 12 h.Supernatant is abandoned in suction, is then respectively adding the testing compound of 200 μ L various concentrations, while sets positive control drug group With blanc cell control group, every group sets 4 multiple holes.After cultivating 72 h, the 5 mg/mL μ L of MTT 20 are added / hole, continue to abandon supernatant after cultivating 4 h, add the μ L of DMSO 150/hole, vibrate 10 min in microoscillator, will try Agent control zeroing, the OD values of cell controls group and each medicine group are measured at 550 nm wavelength with automatic ELIASA, are taken each Class mean, repeat experiment 3 times.Inhibiting rate IR=(1-medicine of each group medicine to cell is calculated with following formula Group OD values/cell controls group OD values) × 100 %, while calculate IC₅₀Value（Experimental result is shown in Table 3）.From Table 3 can be seen that the compound preferably gone out to HEL（Human erythroleukemia cell）, K 562-1（Myelomatosis Cell）, WM9-1（Human melanoma cell）, MDA 231（Breast cancer cell）, TK -6（B lymphoblasts ）, DP17-17（FV-P induction erythroleukemia (mouse cell)）, PC -3（Prostate gland cancer cell）Have relatively good Activity.

External internal antitumor experiment has been carried out to C 1：

Embodiment 12

Influences of 3 days to the cell growths of K 562, specific side are acted on trypan blue cell counting detection C 1 Method cleans tally and cover glass with alcohol, is then gently dried with blotting paper, with Trypsin Induced single-layer culturing cell or Suspended culture cell is collected, individual cells suspension is made, cover glass is covered among the groove of tally two, is gently blown and beaten with suction pipe thin Born of the same parents' suspension, draw a small amount of cell suspension and mixed with equal proportion Trypan Blue liquid, cover glass side refinement born of the same parents are hanged on tally Liquid, under the microscope, the cell number in the block plaid of tally corner is observed with 10 times of object lens, counts the cell not being colored, When cell presses center line, only meter left side and top person, disregard right side and lower section person.

Fig. 5 is influence curve figures of the C 1 to the cell growths of K 562；It can be seen that compound C's 1 is dense Degree shows have good inhibiting effect to the cells of K 562 with being proportionate to the inhibiting rate of cell.

Embodiment 13

Influences of 72 h to the cell cycles of K 562 is acted on Flow cytometry C 1.

Fig. 6 is the influence to the cell cycles of K 562 when C 1 acts on 72 h, it can be seen that C 1 There is obvious influence to the cell cycles of K 562, block in the G2 phases.

Embodiment 14

To the mouse injection Friend virus viruses of firm birth one day, continuously the medicines of C 1 are used after 35 days（ 1 mg / kg ) two weeks, then dissect, survey its spleen weight.Hematocrit value is surveyed with micro-capillary tubes specific volume method, specific method is with anti-freezing Blown out after agent moistening capillary tube inner wall, allow in wall natural air drying or after being dried in 60 DEG C~80 DEG C drying boxes it is stand-by； Take blood：Routine disinfection, cardiac blood is drawn, blood is entered the 2/3 of capillary（About 50 mm）Place；Centrifugation：Use alcohol The molten envelope of lamp or plasticine, paraffin block its end of not sucking blood, then block and are outwards put into special horizontal capillary pipe centrifuge, with 12000 r/min the min of centrifugation 5, red blood cell post and whole blood pillar height degree are measured with graduated scale respectively（Unit mm ）.Its ratio is calculated, that is, draws hematocrit value.

Fig. 7 is C 1（1 mg/kg) influence of the two weeks to infection F-MuLV mouse spleen weights, In general normal spleen weight is 0.1 or so, and illness is then 0.4 or so, from Fig. 5 it can be seen that C 1 is to illness Mouse has apparent therapeutic action；

Fig. 8 is C 1（1 mg/kg) influence of the two weeks to infection F-MuLV mouse red blood cell specific volumes, Hematocrit value normally is 50 %, and ill hematocrit value is 25 % or so, it can be seen that C 1 from figure There is therapeutic action to the Friend virus leukaemia induced.

It is above-mentioned test result indicates that：Indoles simultaneously [3,2-a] carbazole compound or its is pharmaceutically acceptable Salt shows significant inhibitory action to different tumor cell lines, therefore can be used for preparing antineoplastic.The present invention is Develop antineoplastic and provide new chemical entities or lead compound, have for the Chinese conventional medicament of utilization important Meaning.

Claims

1. a kind of indoles simultaneously [3,2-a] carbazole derivates and its salt of generation, it is characterised in that its general structure is formula (1)：

In formula (1)：

R₁For hydrogen, hydroxyl,

R₂For hydrogen, halogen atom, hydroxyl, C1~C4 alkoxies, C1~C4 alkanoyls；

R₃For：

A kind of 2. indoles according to claim 1 simultaneously [3,2-a] carbazole derivates, it is characterised in that signified derivative salt For pharmaceutically acceptable salt hydrochlorate, sulfate, mesylate, tosilate, maleate, fumarate, tartaric acid Salt.

A kind of 3. indoles according to claim 1 simultaneously [3,2-a] carbazole derivates, it is characterized in that R₁Represent hydrogen, Hydroxyl,R₂Represent hydrogen, methoxyl group, chlorine or bromine；R₃RepresentIt is derivative Thing salt is pharmaceutically acceptable salt hydrochlorate.

4. a kind of indoles according to claim 1 or 2 or 3 simultaneously [3,2-a] carbazole derivates, it is characterized in that compound or its Salt is as follows：

A kind of 5. indoles according to claim 4 simultaneously [3,2-a] carbazole derivates, it is characterized in that the compound synthesized C1, C3, C11, C12, C13, C14, C15, C16, C27, C28 or its hydrochloride.

A kind of 6. application of indoles as described in claim 1 or 2 or 3 simultaneously [3,2-a] carbazole derivates, it is characterized in that for making Standby anti-tumor drug.

A kind of 7. application of indoles as claimed in claim 4 simultaneously [3,2-a] carbazole derivates, it is characterized in that anti-swollen for preparing The medicine of knurl.

A kind of 8. application of indoles according to claim 6 simultaneously [3,2-a] carbazole derivates, it is characterized in that anti-for preparing Human erythroleukemia cell HEL, myelogenous leukemia cells K 562-1, human melanoma cell WM9-1, breast cancer cell MDA 231, B lymphoblast TK-6, FV-P induction erythroleukemia mouse cell DP 17-17, what prostate gland cancer cell PC-3 was formed swells The medicine of knurl.

A kind of 9. application of indoles according to claim 7 simultaneously [3,2-a] carbazole derivates, it is characterized in that anti-for preparing Human erythroleukemia cell HEL, myelogenous leukemia cells K 562-1, human melanoma cell WM9-1, breast cancer cell MDA 231, B lymphoblast TK-6, FV-P induction erythroleukemia mouse cell DP 17-17, what prostate gland cancer cell PC-3 was formed swells The medicine of knurl.