CN102153549A

CN102153549A - Gamma carboline compound as well as preparation method and application thereof

Info

Publication number: CN102153549A
Application number: CN201110030648XA
Authority: CN
Inventors: 邓卫平; 刘建文; 刘力锋; 金惠; 安娜; 梁雷; 皮红军; 刘华; 应俊
Original assignee: East China University of Science and Technology
Current assignee: East China University of Science and Technology
Priority date: 2010-12-14
Filing date: 2011-01-21
Publication date: 2011-08-17
Anticipated expiration: 2031-01-21
Also published as: CN102153549B

Abstract

The invention relates to a gamma carboline compound as well as a preparation method and application thereof, in particular relates to a histone deacetylase inhibitor based on gamma carboline as a skeleton as well as a preparation method and application thereof. The inhibitor in the invention is a compound having a structural formula as follows (shown in the specification). According to the invention, the synthesis method is simple, the condition is mild, and the compound has high tumor inhibiting rate and is hopeful to become a non-cytotoxins antineoplastic medicament.

Description

γ carboline compound, preparation method and purposes

Technical field

The present invention relates to medical technical field, be specifically related to a kind of γ carboline compound, preparation method and purposes, be used to prepare NSC 630176, can be used to prepare non-cytotoxicity class anti-tumor drug.

Background technology

In most eukaryotes, DNA and histone are bundled together formation chromatin, further concentrate to form karyomit(e).The reversible acetylize of the terminal specific lysine residue of histone has vital role in regulate gene expression.This reversible adjusting is by two kinds of enzymes that action function is opposite: histone acetyl based transferase (HAT) and histon deacetylase (HDAC) (HDACs) are controlled.Behind the ε amino acid of the specific lysine residue of acetylize, remove its positive charge, therefore reduced the avidity between electronegative DNA and the histone.This has quickened the close of various transcription factors and DNA, activates tumor suppressor gene.Therefore, in most cases acetylation of histone is strengthened transhipment and dna methylase inhibitor inhibition transhipment.

According to the zymic homologous relationship, people source histon deacetylase (HDAC) can be able to be divided into four classes: I and II class is Zn ²⁺Dependent form, the III class is a NAD dependent form, and IV class (including only HDAC11) had both had the character that HDAC I also has HDAC II.The activity of inhibition of histone deacetylase raises acetylizad histone level, can bring out the reaction of a series of specific cells, and for example apoptosis, differentiation and increment inhibition etc. show that histon deacetylase (HDAC) is a potential cancer therapy target.

NSC 630176 has been used to study their treatment tumor effect.Nowadays, comprise short chain fatty acid, cyclic tetrapeptide and hydroximic acid each interior serial inhibitor entered clinical before or clinical study.SAHA and FK228 (Drugs of Today, 2009,45,787) successively obtained drugs approved by FDA treatment epidermis T lymphocyte tumour at 2006 and 2009, confirm that the inhibition of histone deacetylase is the available strategy of treatment cancer.Except cancer therapy, NSC 630176 also can be used for treating inflammation, nerve degenerative diseases (Current Pharmaceutical Design, 2009,15,3940), cardiovascular disorder (Pharmacol.Res.2010,62,3) etc.

HDLP (Nature, 1999,401,188) and HDAC 8 crystalline structure disclose, Zn ²⁺Be positioned at not only narrow but also dark active pocket bottom, with the carbonyl and the hydroxyl generation sequestering action of hydroximic acid.The NSC 630176 pharmacophore feature that has been found that comprises with the lower section: metal-chelating district, link zone and surperficial cog region.The mechanism of action of illustrating to designing new hdac inhibitor and research inhibitor of the discovery of this crystalline structure and pharmacophore feature provides the foundation.

Although had more hdac inhibitor to be synthesized and to have applied for patent (Expert Opinion on Therapeutic Patents at present; 2009; 19,1727), but from present clinical result; its antitumor curative effect is still waiting to improve; drug effect is better, and side effect is littler, and the better compound of pharmacokinetic properties still remains to be found; also need more alternative NSC 630176 clinically, the present invention comes therefrom.

Contain the female ring of carboline (CN 101175754A, Biorg.Med.Chem.2008,16,4908) in many medicines structures, report less relatively but the carboline compounds is used for the research of NSC 630176.Butler in 2010; K.V etc. have reported that the building stone that the carboline ring is used for surperficial cog region carries out the design (J.Am.Chem.Soc.2010 of NSC 630176; 132; 10842); this class inhibitor reaches 15nM to the half inhibition activity of HDAC 6 enzymes, the selectivity of HDAC 6 is compared with HDAC 1 can reach 1000 times.We are synthetic as the design that link zone carries out NSC 630176 with the carboline ring in this patent; designed the compound of a series of novel structures, and it is active and to the enzyme inhibition activity of histon deacetylase (HDAC) to have tested its vitro inhibition to tumour cell.

Summary of the invention

The purpose of this invention is to provide a kind of γ carboline compound, is a kind of NSC 630176 of γ carboline skeleton furtherly.

Purpose of the present invention also provides a kind of intermediate of synthetic above-claimed cpd.

Purpose of the present invention also provides a kind of preparation method of above-mentioned γ carboline compound.

Another object of the present invention provides a kind of purposes of above-mentioned γ carboline compound, is used to prepare NSC 630176, owing to have the tumors of higher inhibiting rate, being expected preparation becomes non-cytotoxicity class anti-tumor drug.

γ carboline compound of the present invention has following structural formula:

Wherein:

R ¹Be C _1～6Alkyl, C _2～6Thiazolinyl, C _2～6Alkynyl, C _6～10The C that aryl replaces _1～4Alkyl, C _6～10The C that heteroaryl replaces _1～4Alkyl ,-C (O)-C _6～10Aryl ,-C (O)-C _1～2Alkylidene group-C _6～10Aryl ,-S (O) ₂-C _1～6Alkyl ,-S (O) ₂-C _6～10Aryl; Wherein said C _6～10Aryl and C _6～10Heteroaryl can be replaced by one or more substituting groups, and described substituting group comprises: hydrogen, carboxyl, halogen, C ₁～C ₄Alkyl, C ₁～C ₄Alkoxyl group, amino, hydroxyl, nitro or cyano group; C _6～10Heteroaryl is meant and contains 1～2 N, S or the heteroatomic C of O _6～10Aryl;

L ₁Expression C ₁～C ₄Alkylidene group;

R ², R ³Be C ₁-C ₆Alkyl, C ₁-C ₆The assorted alkyl that contains N, S, O atom, each other can be identical or inequality, or each other in encircling into key.

The invention provides a kind of synthetic key intermediate, structural formula such as following as preceding γ carboline compound:

R wherein ¹, R ², R ³, L ₁Definition described as claim 1, R ⁴Expression C ₁～C ₅Alkyl.

Compound of the present invention realizes that by the following method typical reaction formula 1 is as follows:

The Fischer prepared in reaction takes place by para-bromophenyl-hydrazine hydrochloride and 4-piperidone hydrochloride to obtain (J.Heterocyclic Chem in the tetrahydrochysene γ carboline I-1 that 8 bromines of the initial raw material of the prepared compound of the present invention replace under 90 ℃ of reflux conditionss in the saturated hydrogen chloride solution of alcoholic acid, 2006,43,571).To I-1 at sodium bicarbonate (NaHCO ₃) make alkali, methyl alcohol is done under the condition of solvent the N atom is carried out obtaining I-2 after the protection of Boc acid anhydrides.Compound I-2 again with methyl acrylate take place Heck react Compound I-3a, the solvent that this step is used is a dimethyl formamide, acid binding agent is N; the N-diisopropylethylamine; catalyzer is palladium, triphenylphosphine, and reaction needs to carry out under 110 ℃ of conditions under nitrogen protection.Make alkali at sodium hydroxide, Tetrabutyl amonium bromide is done under the phase-transfer catalyst condition and to be introduced the alkane hydrogen that bromine replaces get Compound I-4a on indole ring, obtains Compound I-5a remove the Boc protecting group under acidic conditions after.Make solvent at acetonitrile; salt of wormwood is done under the condition of alkali; on the N atom, introduce the benzyl that replaces; acyl group; alkylsulfonyl and alkyl etc. get Compound I-6a～m; the reduction amination reaction also can be taken place by the aldehyde of I-5a and replacement and obtain in I-6a～m in the presence of reductive agent, reductive agent is sodium triacetoxy borohydride, sodium cyanoborohydride or sodium borohydride.I-6a～m does under the condition of alkali and various primary amine or secondary amine R at salt of wormwood again ²R ³Nucleophilic substitution reaction takes place under 90 ℃ of reflux conditionss NH introduces various substituted amidos and prepares ester class key intermediate I-7a～x.Ester class intermediate is made alkali at potassium hydroxide, and methyl alcohol is done the following 70 ℃ of reflux generation hydrolysis reaction of solvent condition and obtained corresponding acid, and the solvent that need adopt during acid extraction is first-selected to be ethyl acetate.0 ℃ of nitrogen protection, triethylamine is made alkali, tetrahydrofuran (THF) is done under the solvent condition, the azanol room temperature reaction that acid and Vinyl chloroformate reaction obtain mixing behind the acid anhydride with the tetrahydropyrans protection again obtains Compound I-9a～x, and Compound I-9a～x also can be obtained by azanol condensation under the condition of condensing agent existence of acid and tetrahydropyrans protection; (hydrogenchloride/ethyl acetate solution) removes the tetrahydropyrans protecting group and obtains target compound I-10a～x under acidic conditions.

Preparation method of the present invention further describes as follows:

1) the tetrahydrochysene γ carboline I-1 that the Fischer reaction obtained 8 bromines replacements in 3～5 hours takes place in para-bromophenyl-hydrazine hydrochloride and 4-piperidone hydrochloride under 80～100 ℃ of conditions in the saturated hydrogen chloride solution of alcoholic acid; The mol ratio of described para-bromophenyl-hydrazine hydrochloride and 4-piperidone hydrochloride is 1: 0.8～1.2;

2) in water or organic solvent, alkali exists down, under 20～90 ℃ the N atom of Compound I-1 is carried out protective reaction and obtains Compound I-2 in 0.5～4 hour; Protecting group reagent first-selection is the Boc acid anhydrides; The mol ratio of Compound I-1, alkali and Boc acid anhydrides is 1: 1～2.2: 0.9～1.2; Described Boc represents tertbutyloxycarbonyl;

3) in the presence of organic solvent, acid binding agent and palladium catalyst, Compound I-2 and CH ₂CHCOOR ⁴The Heck linked reaction taking place 8～10 hours at 90～120 ℃, obtains Compound I-3; Palladium catalyst is the mixture of tetra-triphenylphosphine palladium, palladium and triphenylphosphine or the mixture of Palladous chloride and triphenylphosphine, described Compound I-2, CH ₂CHCOOR ⁴, acid binding agent and palladium, triphenylphosphine mol ratio be 1: 2～10: 1～3: 0.05～0.15: 0.15～0.45; I-2, CH ₂CHCOOR ⁴, acid binding agent and tetra-triphenylphosphine palladium mol ratio be 1: 2～10: 1～3: 0.05～0.15; R ³Definition as shown in the right 2;

4) in the presence of alkali and phase-transfer catalyst, Compound I-3 and excessive XL ₁X gets Compound I-4 at 40～60 ℃ of reaction 12～36h, and alkali is sodium hydroxide, the aqueous solution of potassium hydroxide, and concentration is 2～12M; Described Compound I-3, XL ₁The mol ratio of X, alkali and phase-transfer catalyst is 1: 2～10: 2～10: 0.1～1; Described phase-transfer catalyst is quaternary ammonium salt and quaternary alkylphosphonium salt;

5) at 20～60 ℃, Compound I-4 is dissolved in the organic solvent, removes the reaction of Boc protecting group and obtained Compound I-5 in 0.5～4 hour in the presence of acid reagent, and acid reagent comprises trifluoroacetic acid, hydrochloric acid, sulfur oxychloride or tosic acid; The mol ratio of Compound I-4 and acid reagent is 1: 2～10;

6) at 0～40 ℃, organic solvent and alkali exist down, Compound I-5 and R ¹X reaction 2～24h gets Compound I-6; X is a halogen, R ¹, L ₁Definition described as claim 1; I-5, R ¹L ₁The mol ratio of X 1: 1～2;

Perhaps Compound I-5 and R ¹CHO got Compound I-6 in 6～36 hours with reductive agent generation reductive amination process in organic solvent; Compound I-5, R ¹The mol ratio of CHO and reductive agent is 1: 1～1.5: 1～3; Described reductive agent is sodium triacetoxy borohydride, sodium cyanoborohydride or sodium borohydride;

7) at 40～100 ℃, in the presence of alkali and organic solvent, Compound I-6 and R ²R ³NH reaction 4～24h obtains ester class intermediate compound I-7; R ², R ³Definition described as claim 1; Described Compound I-6, R ²R ³The mol ratio of NH and alkali is 1: 1～5: 1～4;

8) at 20 ℃～100 ℃, ester class intermediate compound I-7 alkaline condition issue unboiled water separate the reaction 2～8h obtain corresponding sour I-8; The mol ratio of described ester class intermediate compound I-7 and alkali is 1: 1～5;

9) under 0 ℃ of protection of inert gas, in the presence of organic solvent and acid binding agent, I-8 and Vinyl chloroformate reaction 0.2～0.5h obtain mixing acid anhydride; The azanol room temperature reaction 2～6h that adds the tetrahydropyrans protection again obtains Compound I-9; I-8, acid binding agent, Vinyl chloroformate, tetrahydropyrans mol ratio are 1: 1～3: 1～3: 1～5;

Perhaps in the organic solvent of 20～40 ℃ of temperature, the azanol of Compound I-8 and tetrahydropyrans protection reacts 12～36h condensation and obtains Compound I-9 in the presence of condensing agent, described condensing agent is N, N '-dicyclohexylcarbodiimide, 1-ethyl-3-(3-dimethylamine propyl) carbodiimide hydrochloride, N-hydroxy benzo triazole; The azanol of I-8, tetrahydropyrans protection, the mol ratio of condensing agent are 1: 1～4: 1～4;

10) at 0～60 ℃, organic solvent and acid exist down, and Compound I-9 reaction removed the tetrahydropyrans protecting group in 0.5～2 hour and obtains target compound I-10; Acid reagent is hydrochloric acid, acetic acid, boric acid or boron trifluoride diethyl etherate; The mol ratio 1: 1～10 of I-9, acid;

The structural formula of Compound I-1, I-2, I-3, I-4, I-5, I-6, I-8 and I-9 is as follows respectively:

Wherein X is a halogen; R ¹, R ², R ³, R ⁴And L ₁As previously mentioned; Shown in the key intermediate of the same γ carboline compound of Compound I-7 structural formula, Compound I-10 structural formula is shown in the γ carboline compound.

Typical compound of the present invention is as shown in table 1.

Table 1 Compound I-10a～x table look-up

γ carboline compound of the present invention is a kind of NSC 630176 of γ carboline skeleton, has investigated the application of carboline histone deacetylases inhibitor I-10a～x in the preparation antitumor drug.Preliminary in-vitro screening finds that they are to various tumor cell strains; comprise human breast cancer cell strain (MCF-7), human lung carcinoma cell line (A549); human colon cancer cell strain (HCT116), human cervical carcinoma cell strain (HeLa) and human leukemia cell line (K562) all have inhibition increment effect (shown in the table 2) in various degree, and IC is measured in the effect of carboline histone deacetylases inhibitor respectively after 48 hours ₅₀Value, most compounds obviously are better than positive control medicine SAHA (popular name: vorinostat) or suitable with it to the restraining effect of 5 kinds of tumour cells.Compound I-10b wherein, I-10e, I-10g, I-10h, I-10l is for the IC of various tumour cells ₅₀Value obviously is better than SAHA, is example with Compound I-10h, and it is that 30 times of SAHA reach 0.152 μ M to the HCT116 cell inhibiting activity, reaches about 2～10 times of SAHA for the activity of other tumour cell.Also tested part of compounds among I-10a～t to the enzyme inhibition activity of HDACs and HDAC 1 hypotype.The result shows (shown in the table 3), this series compound for the enzyme inhibition activity of HDACs and HDAC 1 and SAHA quite or a little less than SAHA.With Compound I-10h is example, and its half-inhibition concentration to HDACs reaches 0.063 μ M, reaches 0.021 μ M for the half-inhibition concentration of HDAC 1.Further pharmacological experiment to Compound I-10l shows, this series compound can be induced the apoptosis of neuroblastoma SHSY5Y, in the G1 phase, the transplanted tumor cell animal model of mouse proves that also this series compound can have the tumors of higher inhibiting rate with cell-cycle arrest.γ carboline compound of the present invention is expected to prepare and becomes non-cytotoxicity class and the cytodifferentiation disease medicament relevant with propagation.

What the present invention is directed to is meant solid tumor or non-solid tumor, comprises relative diseases such as metastases, comprises malignant melanoma, malignant lymphoma, Alimentary tumour (cancer of the stomach, intestinal cancer, liver cancer, carcinoma of gallbladder, cancer of bile ducts, carcinoma of the pancreas), lung cancer, breast cancer, carcinoma of testis, ovarian cancer, uterus carcinoma, prostate cancer, maxilla cancer, tongue cancer, oral carcinoma, laryngocarcinoma, thyroid carcinoma, brain tumor, each sarcoid, osteosarcoma, leukemia, nervous system neoplasm, tumor of bladder, skin carcinoma, skin accessory organ's cancer, metastatic carcinoma of skin and medically affiliated all kinds of tumours and transfer thereof.

Description of drawings

Fig. 1 Hoechst Study on dyeing different concns I-10l is to the influence of SHSY5Y form.

Fig. 2 different concns I-10l effect 48h is to the influence of the cell cycle of SHSY5Y.

Figure 31 μ M I-10l effect different time is to the influence of the cell cycle of SHSY5Y.

Fig. 4 different concns I-10l effect 48h is to the influence of SHSY5Y apoptosis induction.

Figure 51 μ M I-10l effect different time influences the SHSY5Y apoptosis induction.

Nomenclature

Fig. 2, the G0-G1 among Fig. 3, S, G2-S represent the intermitosis 1 in the cell division cycle respectively, DNA synthesis phase and intermitosis 2. ^*Expression p＜0.05, ^*Expression p＜0.01.All experiments are all carried out more than three times and three times.

Fig. 4, the PapE among Fig. 5, PapL, total represent the early apoptosis rate respectively, late period apoptosis rate and total apoptosis rate. ^*Expression p＜0.05, ^*Expression p＜0.01, all experiments are all carried out more than three times and three times.

Embodiment

The present invention is further described in conjunction with the embodiments.Following embodiment is that explanation is of the present invention, rather than limits content of the present invention by any way.

The preparation of embodiment 1 Compound I-10a～v

Step 1

8-bromo-2,3,4, the preparation of 5-tetrahydrochysene-1H-pyridine [4,3-b] indole hydrochlorides (I-1)

Para-bromophenyl-hydrazine hydrochloride 22.35g (0.1mol) and 4-piperidone hydrochloride 15.36g (0.1mol) are dissolved in the saturated hydrogen chloride solution of 400mL alcoholic acid, and the adularescent solid is separated out behind the back flow reaction 3h.Be cooled to room temperature, the white solid of separating out is filtered, the filter cake absolute ethanol washing obtains 22.2g white solid product I-1, yield 77% after the vacuum drying oven drying.

¹H?NMR(400MHz，DMSO)：δ(ppm)：11.51(s，1H)，9.74(s，2H)，7.69(s，1H)，7.31(d，J＝8.6Hz，1H)，7.19(dd，J＝8.6，1.6Hz，1H)，4.25(s，2H)，3.42(t，J＝5.3Hz，2H)，3.04(t，J＝5.3Hz，2H).

¹³C?NMR(100MHz，DMSO)δ135.01，132.93，127.21，124.08，120.52，113.52，111.90，102.02，41.04，40.10，20.54.

Step 2

8-bromo-3, the preparation of 4-dihydro-1H-pyridine [4,3-b] indoles-2 (5H)-carboxylic acid tert-butyl ester (I-2)

Under the room temperature 28.8g (0.1mol) Compound I-1 is dissolved in the methanol solution of 350mL, adds 26.2g (0.12mol) tert-Butyl dicarbonate (Boc acid anhydrides) again behind adding 20.2g (0.24mol) sodium bicarbonate in batches, 60 ℃ are reacted 1h down.Reaction finishes, and the methyl alcohol decompression is spin-dried for the back uses dichloromethane extraction, combining extraction liquid, the saturated common salt water washing, anhydrous sodium sulfate drying is crossed the silica gel short column after concentrating and is obtained faint yellow Compound I-229.9g, yield 85%, M.p.170-171 ℃.

¹H?NMR(400MHz，CDCl ₃)：δ(ppm)：7.95(brs，1H)，7.57(s，1H)，7.23(d，J＝8.6Hz，1H)，7.17(d，J＝8.6Hz，1H)，4.58(s，2H)，3.86-3.76(m，2H)，2.82(t，J＝4.6Hz，2H)，1.51(s，9H).

¹³C?NMR(100MHz，DMSO)δ154.71，134.99，127.39，123.43，120.05，113.28，111.55，79.42，28.57.

Step 3

(E)-and 8-(1-methyl acrylate base)-3, the preparation of 4-dihydro-1H-pyridine [4,3-b] indoles-2 (5H)-carboxylic acid tert-butyl ester (I-3a)

Under the nitrogen protection; to being dissolved with 14.05g (0.04mol) Compound I-2; 0.896g (4mmol) palladium adds 14.4mL (0.08mol) diisopropyl ethyl amine in 250mL dimethyl formamide (DMF) solution of 3.15g (12mmol) triphenylphosphine, 36mL (0.4mol) methyl acrylate.Reaction solution is reacted 8h down at 110 ℃, remove unnecessary solvent under reduced pressure after reaction finishes, column chromatography obtains yellow solid Compound I-3a12.0g, yield 84%, M.p.207-208 ℃.

¹H?NMR(400MHz，CDCl ₃)：δ(ppm)：8.13(s，1H)，7.83(d，J＝15.9Hz，1H)，7.61(s，1H)，7.37(dd，J＝8.5，1.3Hz，1H)，7.29(d，J＝8.4Hz，1H)，6.41(d，J＝15.9Hz，1H)，4.64(s，2H)，3.86-3.77(m，5H)，2.88-2.77(m，2H)，1.51(s，9H).

¹³C?NMR(100MHz，DMSO)δ167.62，154.73，147.19，137.73，125.92，125.27，121.33，119.59，114.08，111.87，79.42，51.59，28.56.

Step 4

(E)-and 5-(2-bromotrifluoromethane)-8-(1-methyl acrylate base)-3, the preparation of 4-dihydro-1H-pyridine [4,3-b] indoles-2 (5H)-carboxylic acid tert-butyl ester I-4a (n=1)

3.2g (9mmol) Compound I-3 is dissolved in the 30mL methylene dichloride, adds aqueous sodium hydroxide solution, 14mL glycol dibromide and the 150mg Tetrabutyl amonium bromide of 14mL 9M.60 ℃ of back flow reaction 36h of reaction solution postcooling to room temperature, is used dichloromethane extraction, merge organic phase, the saturated common salt water washing, column chromatography obtains 2.32g faint yellow solid Compound I-4 (n=1), yield 56%, M.p.142-144 ℃ behind the anhydrous sodium sulfate drying.

¹H?NMR(400MHz，CDCl ₃)：δ(ppm)：7.83(d，J＝15.9Hz，1H)，7.63(s，1H)，7.43(d，J＝8.6Hz，1H)，7.28(d，J＝8.6Hz，1H)，6.42(d，J＝15.9Hz，1H)，4.64(s，2H)，4.43(t，J＝7.1Hz，2H)，3.89-3.83(m，2H)，3.81(s，3H)，3.57(t，J＝7.1Hz，2H)，2.88-2.81(m，2H)，1.51(s，9H).

(E)-and 5-(3-bromopropyl)-8-(1-methyl acrylate base)-3, the preparation of 4-dihydro-1H-pyridine [4,3-b] indoles-2 (5H)-carboxylic acid tert-butyl ester I-4a (n=2)

The preparation method of reference compound I-4a (n=1) replaces with 1 with the raw material glycol dibromide, 3-dibromopropane, yellow oily Compound I-4a (n=2), yield 62%.

¹H?NMR(400MHz，CDCl ₃)：δ(ppm)：7.85(d，J＝15.9Hz，1H)，7.64(d，J＝1.2Hz，1H)，7.44(dd，J＝8.5，1.2Hz，1H)，7.36(d，J＝8.5Hz，1H)，6.44(d，J＝15.9Hz，1H)，4.66(s，2H)，4.24(t，J＝6.7Hz，2H)，3.87(t，J＝4.8Hz，2H)，3.83(s，3H)，3.37(t，J＝6.7Hz，2H)，2.86(t，J＝4.8Hz，2H)，2.35-2.29(m，2H)，1.54(s，9H).

Step 5

(E)-and 5-(2-bromotrifluoromethane)-8-(1-methyl acrylate base)-2,3,4, the preparation of 5-tetrahydrochysene-1H-pyridine [4,3-b] indole hydrochloride I-5a (n=1)

2.32g (5mmol) Compound I-4a (n=1) is dissolved in the 20mL methylene dichloride, adds hydrogenchloride/ethyl acetate solution of 10mL6M, stirring at room reaction 0.5h has a large amount of brown solid to separate out.With the solid filtering of separating out, filter cake obtains 1.95g brown solid I-5a (n=1), yield 98% with the petroleum ether after drying.

¹³C?NMR(100MHz，CDCl ₃)δ168.17，146.27，136.98，134.15，126.23，125.99，121.35，119.15，114.70，109.28，108.63，51.56，44.44，42.66，41.51，29.48，22.39.

(E)-and 5-(3-bromopropyl)-8-(1-methyl acrylate base)-2,3,4, the preparation of 5-tetrahydrochysene-1H-pyridine [4,3-b] indole hydrochloride I-5a (n=2)

The preparation method of reference compound I-5a (n=1) replaces with I-4a (n=2) with raw material I-4a (n=1), obtains white-yellowish solid I-5a (n=2), yield 98%.

Step 6

(E)-preparation of 3-(5-(2-bromotrifluoromethane)-2-benzyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl) methyl acrylate (I-6l)

800mg (2mmol) Compound I-5a (n=1) and 829mg (6mmol) salt of wormwood are dissolved in the 20mL acetonitrile solution, slowly drip 0.25mL (2.1mmol) bromobenzyl.Behind the room temperature reaction 2h,, merge organic phase, saturated common salt water washing, concentrating under reduced pressure behind the anhydrous sodium sulfate drying with the reaction solution dichloromethane extraction.The concentrated solution column chromatography is obtained 707mg weak yellow foam shape solid product I-6l, yield 78%, M.p.117-118 ℃.

¹H?NMR(400MHz，CDCl ₃)：δ(ppm)：7.80(d，J＝15.9Hz，1H)，7.54(s，1H)，7.45-7.23(m，7H)，6.39(d，J＝15.9Hz，1H)，4.40(t，J＝7.3Hz，2H)，3.81(s，2H)，3.79(s，3H)，3.71(s，2H)，3.54(t，J＝7.3Hz，2H)，2.93(t，J＝5.5Hz，2H)，2.87(t，J＝5.5Hz，2H).

¹³C?NMR(100MHz，CDCl ₃)δ168.02，146.58，138.34，137.39，134.72，129.17，128.45，127.32，126.39，126.10，121.11，119.18，114.51，109.80，109.15，62.33，51.52，50.00，49.50，44.68，29.13，22.92.

(E)-preparation of 3-(5-(2-bromotrifluoromethane)-2-o-cyanobenzyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl) methyl acrylate (I-6a)

The preparation method of reference compound I-6l replaces the raw material bromobenzyl with adjacent cyano-benzyl bromide, faint yellow oily thing I-6a, yield 76%.

(E)-preparation of 3-(5-(2-bromotrifluoromethane)-2-to nitrobenzyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl) methyl acrylate (I-6b)

The preparation method of reference compound I-6l uses the raw material bromobenzyl to the replacement of nitro bromobenzyl, faint yellow oily thing I-6b, yield 76%.

¹H?NMR(400MHz，CDCl ₃)δ8.23(d，J＝8.5Hz，2H)，7.81(d，J＝15.9Hz，1H)，7.62(d，J＝8.5Hz，2H)，7.55(s，1H)，7.41(d，J＝8.4Hz，1H)，7.30-7.28(m，1H)，6.40(d，J＝15.9Hz，1H)，4.43(t，J＝7.1Hz，2H)，3.91(s，2H)，3.81(s，3H)，3.74(s，2H)，3.59(t，J＝7.1Hz，2H)，3.00-2.88(m，4H).

(e)-preparation of 3-(5-(2-bromotrifluoromethane)-2-to bromobenzyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl) methyl acrylate (I-6c)

Under the nitrogen protection, with 100mg (0.25mmol) Compound I-5 (n=1), 69mg (0.375mmol) p-bromobenzaldehyde and 106mg (0.5mmol) sodium triacetoxy borohydride join in the reaction tubes of 10mL, add the 5mL methylene dichloride again, room temperature reaction 24h.After reaction finishes reaction solution is added the water dichloromethane extraction, merge organic phase, the saturated common salt water washing, column chromatography obtains the faint yellow oily thing of 86mg I-6c, yield 65% behind the anhydrous sodium sulfate drying.

(E)-preparation of 3-(5-(2-bromotrifluoromethane)-2-α methylnaphthalene-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl) methyl acrylate (I-6d)

The preparation method of reference compound I-6l replaces white solid I-6d, yield 80% with the raw material bromobenzyl with α brooethyl naphthalene.

¹H?NMR(400MHz，CDCl ₃)：δ(ppm)：8.35-8.33(m，1H)，7.87-7.77(m，3H)，7.54-7.43(m，5H)，7.37(dd，J＝8.5，1.0Hz，1H)，7.24(d，J＝8.1Hz，1H)，6.38(d，J＝15.9Hz，1H)，4.37(t，J＝7.3Hz，2H)，4.20(s，2H)，3.80(s，2H)，3.79(s，3H)，3.52(t，J＝7.3Hz，2H)，2.99(t，J＝5.5Hz，2H)，2.81(t，J＝5.2Hz，2H).

(E)-preparation of 3-(5-(2-bromotrifluoromethane)-2-β methylnaphthalene-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl) methyl acrylate (I-6e)

The preparation method of reference compound I-6l replaces faint yellow oily thing I-6e, yield 82% with the raw material bromobenzyl with β brooethyl naphthalene.

(E)-preparation of 3-(5-(2-bromotrifluoromethane)-2-to methoxy-benzyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl) methyl acrylate (I-6f)

The preparation method of reference compound I-6l uses the raw material bromobenzyl to the replacement of methoxyl group bromobenzyl, yellow oil I-6f, yield 49%.

(E)-preparation of 3-(5-(2-bromotrifluoromethane)-2-p-toluenesulfonyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl) methyl acrylate (I-6g)

Under the room temperature condition, 300mg (0.75mmol) I-5 (n=1) is dissolved in the 25mL methylene dichloride, adds 0.31mL (2.25mmol) triethylamine, add 286mg (1.5mmol) Tosyl chloride more in batches.Behind the reaction 1h,, use dichloromethane extraction, merge organic phase the reaction solution thin up, the saturated common salt water washing, anhydrous sodium sulfate drying, column chromatography behind the concentrating under reduced pressure is received solid phase prod I-6g 289mg, yield 75%.

(E)-preparation of 3-(5-(2-bromotrifluoromethane)-2-n-propyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl) methyl acrylate (I-6h)

The preparation method of reference compound I-6l replaces the raw material bromobenzyl with iodopropane, faint yellow oily thing I-6h, yield 32%.

¹H?NMR(400MHz，CDCl ₃)：δ(ppm)：7.82(d，J＝15.9Hz，1H)，7.58(s，1H)，7.39(d，J＝7.8Hz，1H)，7.26(d，J＝7.8Hz，1H)，6.41(d，J＝15.9Hz，1H)，4.40(t，J＝7.2Hz，2H)，3.80(s，3H)，3.70(s，2H)，3.53(t，J＝7.2Hz，2H)，2.95-2.83(m，4H)，2.60(t，J＝7.6Hz，2H)，1.70-1.66(m，4H)，0.98(t，J＝7.3Hz，3H).

(E)-preparation of 3-(5-(2-bromotrifluoromethane)-2-methylsulfonyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl) methyl acrylate (I-6i)

The preparation method of reference compound I-6g replaces with methylsulfonyl chloride with Tosyl chloride, yellow oil I-6i, yield 98%.

(E)-preparation of 3-(5-(2-bromotrifluoromethane)-2-benzoyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl) methyl acrylate (I-6j)

The preparation method of reference compound I-6g replaces with Benzoyl chloride with Tosyl chloride, faint yellow solid I-6j, yield 91%.

¹H?NMR(400MHz，CDCl ₃)δ7.82-7.64(m，1H)，7.48-7.34(m，7H)，7.24(d，J＝8.6Hz，1H)，6.49-6.23(m，1H)，4.92(s，1H)，4.63(s，1H)，4.40(t，J＝7.0Hz，3H)，4.15(s，1H)，3.80-3.70(m，4H)，3.55(t，J＝7.0Hz，3H)，2.99-2.83(m，2H).

(E)-preparation of 3-(5-(2-bromotrifluoromethane)-2-phenylacetyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl) methyl acrylate (I-6k)

The preparation method of reference compound I-6g replaces with phenyllacetyl chloride with Tosyl chloride, faint yellow solid I-6k, yield 61%, M.p.153-154 ℃.

(E)-preparation of 3-(5-(3-bromopropyl)-2-benzyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl) methyl acrylate (I-6m)

The preparation method of reference compound I-6l replaces with I-5 (n=2) with raw material I-5 (n=1), pale brown look oily matter I-6m, yield 62%.

¹H?NMR(400MHz，CDCl ₃)：δ(ppm)：7.80(d，J＝15.9Hz，1H)，7.53(s，1H)，7.42(d，J＝7.0Hz，2H)，7.35(t，J＝7.3Hz，3H)，7.30(d，J＝8.0Hz，2H)，6.38(d，J＝15.9Hz，1H)，4.18(t，J＝6.7Hz，2H)，3.80(s，2H)，3.79(s，3H)，3.71(s，2H)，3.35(t，J＝6.7Hz，2H)，2.91(t，J＝5.0Hz，2H)，2.85(t，J＝5.0Hz，2H)，2.32-2.25(m，2H).

Step 7

(E)-preparation of 3-(5-(2-(piperidines-1-yl) ethyl)-2-benzyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl) methyl acrylate (I-7l)

With 707mg (1.56mmol) Compound I-6l, 647mg (4.68mmol) salt of wormwood, 310mg (1.87mmol) potassiumiodide joins in the round-bottomed flask of 50mL, adds 20mL acetonitrile and 3mL (31.2mmol) piperidines again, back flow reaction 6h.After reaction finishes reaction solution is cooled to room temperature, uses dichloromethane extraction, merge organic phase, the saturated common salt water washing, column chromatography obtains 604m g weak yellow foam solid I-7l, yield 85% behind the anhydrous sodium sulfate drying.

¹H?NMR(400MHz，CDCl ₃)：δ(ppm)：7.80(d，J＝15.9Hz，1H)，7.52(s，1H)，7.44-7.39(m，J＝7.4Hz，2H)，7.37-7.31(m，3H)，7.30-7.21(m，J＝19.3，7.7Hz，2H)，6.37(d，J＝15.9Hz，1H)，4.10(t，J＝7.5Hz，2H)，3.78(s，2H)，3.78(s，3H)，3.70(s，2H)，2.89(t，J＝5.2Hz，2H)，2.85-2.79(t，J＝5.2Hz，2H)，2.55(t，J＝7.5Hz，2H)，2.48-2.38(m，4H)，1.64-1.53(m，4H)，1.47-1.37(m，2H).

¹³C?NMR(100MHz，CDCl ₃)δ168.07，146.96，138.63，137.65，135.14，129.12，128.40，127.22，126.16，125.48，120.59，119.17，113.94，109.34，109.02，62.42，58.51，55.08，51.42，50.16，49.72，41.24，26.02，24.26，22.97.

(E)-preparation of 3-(5-(2-(piperidines-1-yl) ethyl)-2-o-cyanobenzyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl) methyl acrylate (I-7a)

The preparation method of reference compound I-7l, I-6l replaces with I-6a with raw material, yellow oily liquid I-7a, yield 64%.

¹H?NMR(500MHz，CDCl ₃)δ7.80(d，J＝15.9Hz，1H)，7.70-7.65(m，1H)，7.61-7.55(m，1H)，7.42-7.34(m，2H)，7.29-7.26(m，2H)，6.38(d，J＝15.9Hz，1H)，4.14(t，J＝7.5Hz，2H)，4.00(s，2H)，3.79(s，3H)，3.75(s，2H)，3.01(t，J＝5.4Hz，1H)，2.89(t，J＝5.4Hz，2H)，2.58(t，J＝7.5Hz，2H)，2.49-2.38(m，4H)，1.62-1.56(m，4H)，1.49-1.41(m，2H).

¹³C?NMR(100MHz，CDCl ₃)δ168.01，146.83，142.81，137.58，135.03，132.91，132.72，129.93，127.69，125.99，125.50，120.66，119.01，117.83，113.98，112.91，109.39，108.56，59.87，58.45，55.06，51.41，50.50，49.44，41.28，25.98，24.21，22.89.

(E)-preparation of 3-(5-(2-(piperidines-1-yl) ethyl)-2-to nitro base benzyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl) methyl acrylate (I-7b)

The preparation method of reference compound I-7l, I-6l replaces with I-6b with raw material, yellow oil I-7b, yield 73%.

¹H?NMR(400MHz，CDCl ₃)δ8.23(d，J＝8.5Hz，2H)，7.82(d，J＝15.9Hz，1H)，7.63(d，J＝8.5Hz，2H)，7.54(s，1H)，7.39(d，J＝8.5Hz，1H)，7.29(d，J＝7.9Hz，1H)，6.39(d，J＝15.9Hz，1H)，4.16(t，J＝7.4Hz，2H)，3.91(s，2H)，3.80(s，3H)，3.75(s，2H)，2.98-2.86(m，4H)，2.64-2.58(m，2H)，2.51-2.42(m，4H)，1.65-1.57(m，4H)，1.50-1.41(m，2H).

¹³C?NMR(100MHz，CDCl ₃)δ168.02，147.22，146.78，146.75，137.60，134.94，129.41，125.97，125.60，123.62，120.75，118.98，114.09，109.42，108.53，61.42，58.50，55.11，51.44，50.38，49.77，41.35，26.00，24.21，22.92.

(E)-preparation of 3-(5-(2-(piperidines-1-yl) ethyl)-2-to bromobenzyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl) methyl acrylate (I-7c)

The preparation method of reference compound I-7l, I-6l replaces with I-6c with raw material, white solid I-7c, yield 65%.

¹H?NMR(500MHz，CDCl ₃)δ7.80(d，J＝15.9Hz，1H)，7.53(s，1H)，7.49-7.45(m，2H)，7.36(d，J＝8.5Hz，1H)，7.32-7.29(m，2H)，7.28-7.24(m，1H)，6.38(d，J＝15.9Hz，1H)，4.13(t，J＝7.5Hz，2H)，3.79(s，3H)，3.74(s，2H)，3.70(s，2H)，2.91-2.87(m，2H)，2.86-2.81(m，2H)，2.57(t，J＝7.5Hz，2H)，2.49-2.38(m，4H)，1.61-1.57(m，4H)，1.47-1.41(m，2H).

¹³C?NMR(100MHz，CDCl ₃)δ167.99，146.86，137.77，137.60，135.04，131.45，130.69，126.07，125.50，120.94，120.64，119.08，113.99，109.37，108.80，61.57，58.43，55.04，51.41，50.14，49.64，41.22，25.97，24.21，22.93.

(E)-preparation of 3-(5-(2-(piperidines-1-yl) ethyl)-2-α methylnaphthalene-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl) methyl acrylate (I-7d)

The preparation method of reference compound I-7l, I-6l replaces with I-6d with raw material, yellow oil I-7d, yield 76%.

¹H?NMR(400MHz，CDCl ₃)δ8.47-8.34(m，1H)，7.91-7.87(m，1H)，7.86(s，1H)，7.83(d，J＝6.0Hz，1H)，7.60-7.46(m，5H)，7.38(d，J＝8.5Hz，1H)，7.31-7.28(m，1H)，6.41(d，J＝15.9Hz，1H)，4.23(s，2H)，4.14(t，J＝7.5Hz，2H)，3.84(s，2H)，3.82(s，3H)，3.01(t，J＝5.5Hz，2H)，2.85(d，J＝5.5Hz，2H)，2.59(t，J＝7.5Hz，2H)，2.49-2.41(m，4H)，1.66-1.53(m，4H)，1.51-1.40(m，2H).

¹³C?NMR(100MHz，CDCl ₃)δ168.16，147.00，137.61，135.35，134.45，133.97，132.65，128.48，128.14，127.42，126.18，125.94，125.70，125.53，125.26，124.84，120.61，119.18，113.98，109.36，109.09，60.45，58.49，55.11，51.48，50.18，50.00，41.28，26.03，24.26，22.97.

(E)-preparation of 3-(5-(2-(piperidines-1-yl) ethyl)-2-β methylnaphthalene-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl) methyl acrylate (I-7e)

The preparation method of reference compound I-7l, I-6l replaces with I-6e with raw material, yellow oil I-7e, yield 53%.

¹H?NMR(400MHz，CDCl ₃)δ7.90-7.79(m，5H)，7.62(d，J＝8.4Hz，1H)，7.55(s，1H)，7.52-7.49(m，2H)，7.38(d，J＝8.5Hz，1H)，7.29(d，J＝4.9Hz，1H)，6.40(d，J＝15.9Hz，2H)，4.16(d，J＝7.4Hz，2H)，3.99(s，2H)，3.81(s，3H)，3.80(s，2H)，2.99(t，J＝5.2Hz，2H)，2.89(t，J＝5.2Hz，2H)，2.65-2.59(m，2H)，2.51-2.43(m，4H)，1.68-1.57(m，4H)，1.52-1.41(m，2H).

(E)-preparation of 3-(5-(2-(piperidines-1-yl) ethyl)-2-to methoxy-benzyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl) methyl acrylate (I-7f)

The preparation method of reference compound I-7l, I-6l replaces with I-6f with raw material, yellow oil I-7f, yield 79%.

¹H?NMR(400MHz，CDCl ₃)：δ(ppm)：7.80(d，J＝15.9Hz，1H)，7.54(s，1H)，7.36-7.32(m，3H)，7.24(d，J＝7.9Hz，1H)，6.89(d，J＝8.5Hz，2H)，6.38(d，J＝15.9Hz，1H)，4.13(t，J＝7.5Hz，2H)，3.82(s，3H)，3.79(s，3H)，3.74(s，2H)，3.69(s，2H)，2.90(t，J＝5.2Hz，2H)，2.84(t，J＝5.2Hz，2H)，2.57(t，J＝7.5Hz，2H)，2.45(m，4H)，1.61-1.57(m，4H)，1.45(m，2H).

(E)-preparation of 3-(5-(2-(piperidines-1-yl) ethyl)-2-p-toluenesulfonyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl) methyl acrylate (I-7g)

The preparation method of reference compound I-7l, I-6l replaces with I-6g with raw material, faint yellow solid I-7g, yield 68%, M.p.90-92C.

¹H?NMR(400MHz，DMSO)δ7.89(s，1H)，7.76-7.70(m，3H)，7.47(d，J＝8.5Hz，1H)，7.44-7.39(m，J＝8.4Hz，3H)，6.56(d，J＝15.9Hz，1H)，4.31(s，2H)，4.11(t，J＝6.4Hz，2H)，3.72(s，3H)，3.47(t，J＝5.3Hz，2H)，2.92-2.85(m，2H)，2.42-2.36(m，5H)，2.35-2.28(m，4H)，1.46-1.39(m，4H)，1.38-1.32(m，2H).

¹³C?NMR(100MHz，CDCl ₃)δ167.82，146.45，143.54，137.43，133.98，133.88，129.69，127.53，125.69，125.31，120.98，118.76，114.28，109.59，105.88，58.27，54.95，51.39，43.48，43.02，41.27，25.93，24.12，22.54，21.43.

(E)-preparation of 3-(5-(2-(piperidines-1-yl) ethyl)-2-n-propyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl) methyl acrylate (I-7h)

The preparation method of reference compound I-7l, I-6l replaces with I-6h with raw material, yellow oil I-7h, yield 71%.

¹H?NMR(400MHz，CDCl ₃)：δ(ppm)：7.82(d，J＝15.9Hz，1H)，7.57(s，1H)，7.36(d，J＝8.1Hz，1H)，7.25(d，J＝8.1Hz，1H)，6.39(d，J＝15.9Hz，1H)，4.12(t，J＝7.3Hz，2H)，3.79(s，3H)，3.70(s，2H)，2.94-2.80(m，4H)，2.61-2.54(m，4H)，2.50-2.36(m，4H)，1.70-1.64(m，2H)，1.63-1.51(m，4H)，1.49-1.37(m，2H)，0.98(t，J＝7.3Hz，3H).

¹³C?NMR(100MHz，CDCl ₃)δ181.93，168.03，146.93，137.62，135.08，126.14，125.38，120.46，119.05，113.82，109.27，108.99，60.06，58.44，55.01，51.35，50.60，49.49，41.16，25.94，24.18，22.92，20.74，12.02.

(E)-preparation of 3-(5-(2-(piperidines-1-yl) ethyl)-2-methylsulfonyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl) methyl acrylate (I-7i)

The preparation method of reference compound I-7l, I-6l replaces with I-6i with raw material, yellow solid I-7i, yield 71%, M.p.172-173 ℃.

¹H?NMR(400MHz，CDCl ₃)：δ(ppm)：7.81(d，J＝15.9Hz，1H)，7.58(s，1H)，7.42(d，J＝8.6Hz，1H)，7.30(d，J＝8.6Hz，1H)，6.41(d，J＝15.9Hz，1H)，4.57(s，2H)，4.13(t，J＝7.0Hz，2H)，3.81(s，3H)，3.75(t，J＝5.7Hz，2H)，2.97(t，J＝5.5Hz，2H)，2.88(s，3H)，2.57(t，J＝7.0Hz，2H)，2.46-2.34(m，4H)，1.59-1.53(m，4H)，1.47-1.38(m，2H).

¹³C?NMR(100MHz，CDCl ₃)δ167.85，146.35，137.45，134.12，125.91，125.33，121.17，118.90，114.50，109.68，106.04，58.41，55.04，51.45，43.17，42.80，41.40，36.38，25.96，24.10，22.55.

(E)-preparation of 3-(5-(2-(Pyrrolidine-1-yl) ethyl)-2-benzoyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl) methyl acrylate (I-7j)

The preparation method of reference compound I-7l, I-6l replaces with I-6j with raw material, and piperidines replaces with Pyrrolidine, light yellow solid I-7j, yield 78%.

¹H?NMR(400MHz，CDCl ₃)δ7.90-7.67(m，1H)，7.55-7.38(m，7H)，7.32(d，J＝8.0Hz，1H)，6.53-6.24(m，1H)，5.04-4.89(m，1H)，4.68(s，1H)，4.25-4.10(m，3H)，3.91-3.61(m，4H)，3.05-2.83(m，2H)，2.78(t，J＝8.0Hz，2H)，2.66-2.48(m，4H)，1.91-1.66(m，4H).

(E)-preparation of 3-(5-(2-(Pyrrolidine-1-yl) ethyl)-2-phenylacetyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl) methyl acrylate (I-7k)

The preparation method of reference compound I-7l, I-6l replaces with I-6k with raw material, and piperidines replaces with Pyrrolidine, pale brown look oily matter I-7k, yield 77%.

(E)-preparation of 3-(5-(3-(piperidines-1-yl) propyl group)-2-benzyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl) methyl acrylate (I-7m)

The preparation method of reference compound I-7l, I-6l replaces with I-6m with raw material, pale yellow oily liquid body I-7m, yield 91%.

¹H?NMR(400MHz，CDCl ₃)：δ(ppm)：7.81(d，J＝15.9Hz，1H)，7.53(s，1H)，7.42(d，J＝7.2Hz，2H)，7.37-7.29(m，5H)，6.38(d，J＝15.9Hz，1H)，4.06(t，J＝6.9Hz，2H)，3.80(s，2H)，3.79(s，3H)，3.73(s，2H)，2.90(t，J＝5.0Hz，2H)，2.85(t，J＝5.0Hz，2H)，2.38-2.27(m，4H)，2.23(t，J＝6.9Hz，2H)，1.92-1.86(m，2H)，1.60-1.56(m，4H)，1.48-1.38(m，2H).

(E)-preparation of 3-(5-(2-(dimethylin) ethyl)-2-benzyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl) methyl acrylate (I-7n)

The preparation method of reference compound I-7l replaces with dimethylamine with the raw material piperidines, pale yellow oily liquid body I-7n, yield 57%.

(E)-preparation of 3-(5-(2-(Pyrrolidine-1-yl) ethyl)-2-benzyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl) methyl acrylate (I-7o)

The preparation method of reference compound I-7l replaces with Pyrrolidine with the raw material piperidines, pale yellow oily liquid body I-7o, yield 73%.

¹H?NMR(400MHz，CDCl ₃)δ7.82(d，J＝15.9Hz，1H)，7.55(s，1H)，7.46-7.42(m，2H)，7.40-7.35(m，3H)，7.33-7.28(m，2H)，6.39(d，J＝15.9Hz，1H)，4.18(t，J＝7.7Hz，2H)，3.82(s，2H)，3.80(s，3H)，3.73(s，2H)，2.96-2.90(m，2H)，2.88-2.83(m，2H)，2.78(t，J＝7.7Hz，2H)，2.65-2.54(m，4H)，1.87-1.79(m，4H).

¹³C?NMR(100MHz，CDCl ₃)δ168.07，146.92，138.60，137.64，135.01，129.12，128.40，127.22，126.15，125.53，120.67，119.18，114.00，109.30，109.15，62.43，55.71，54.53，51.44，50.15，49.71，42.67，23.58，22.97.

(E)-preparation of 3-(5-(2-(morphine quinoline) ethyl)-2-benzyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl) methyl acrylate (I-7p)

The preparation method of reference compound I-7l replaces with the morphine quinoline with the raw material piperidines, pale yellow oily liquid body I-7p, yield 72%, M.p.132-133 ℃.

¹H?NMR(400MHz，CDCl ₃)δ7.82(d，J＝15.9Hz，1H)，7.56(s，1H)，7.47-7.25(m，7H)，，6.40(d，J＝15.9Hz，1H)，4.14(t，J＝7.2Hz，2H)，3.82(s，2H)，3.81(s，3H)，3.75-3.69(m，6H)，2.98-2.83(m，4H)，2.64(t，J＝7.2Hz，2H)，2.49(t，J＝4.5Hz，4H)

(E)-preparation of 3-(5-(2-(diethylin) ethyl)-2-benzyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl) methyl acrylate (I-7q)

The preparation method of reference compound I-7l replaces with diethylamine with the raw material piperidines, pale yellow oily liquid body I-7q, yield 65%.

¹H?NMR(400MHz，CDCl ₃)δ7.80(d，J＝15.9Hz，1H)，7.53(s，1H)，7.44-7.28(m，7H)，6.38(d，J＝15.9Hz，1H)，4.08(t，J＝7.6Hz，2H)，3.81(s，2H)，3.79(s，3H)，3.72(s，2H)，2.94-2.83(m，4H)，2.68(t，J＝7.6Hz，2H)，2.58(q，J＝7.1Hz，4H)，1.02(t，J＝7.1Hz，6H).

¹³C?NMR(101MHz，CDCl ₃)δ168.12，146.94，138.56，137.62，135.05，129.14，128.39，127.22，126.13，125.48，120.65，119.16，113.95，109.23，109.01，62.39，52.71，51.44，50.12，49.71，47.78，42.36，22.98，12.05.

(E)-preparation of 3-(5-(3-(diethylin) propyl group)-2-benzyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl) methyl acrylate (I-7r)

The preparation method of reference compound I-7l, I-6l replaces with I-6m with raw material, and piperidines replaces with diethylamine, yellow oily liquid I-7r, yield 99%.

¹H?NMR(400MHz，CDCl ₃)：δ(ppm)：7.81(d，J＝15.9Hz，1H)，7.54(s，1H)，7.43(d，J＝7.2Hz，2H)，7.38(t，J＝7.2Hz，3H)，7.31-7.29(m，2H)，6.38(d，J＝15.9Hz，1H)，4.05(t，J＝7.3Hz，2H)，3.80(s，2H)，3.79(s，3H)，3.73(s，2H)，2.91(t，J＝5.1Hz，2H)，2.84(t，J＝4.8Hz，2H)，2.50(q，J＝7.1Hz，4H)，2.44(t，J＝6.9Hz，2H)，1.90-1.82(m，2H)，0.99(t，J＝7.1Hz，6H).

(E)-preparation of 3-(5-(3-(Pyrrolidine-1-yl) propyl group)-2-benzyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl) methyl acrylate (I-7s)

The preparation method of reference compound I-7l, I-6l replaces with I-6m with raw material, and piperidines replaces with Pyrrolidine, faint yellow oily thing I-7s, yield 98%

¹H?NMR(400MHz，CDCl ₃)：δ(ppm)：7.81(d，J＝15.9Hz，1H)，7.54(s，1H)，7.43(d，J＝7.3Hz，2H)，7.36(t，J＝7.3Hz，3H)，7.31-7.28(m，2H)，6.38(d，J＝15.9Hz，1H)，4.09(t，J＝7.0Hz，2H)，3.80(s，2H)，3.79(s，3H)，3.73(s，2H)，2.90(t，J＝5.3Hz，2H)，2.84(t，J＝5.3Hz，2H)，2.50-2.44(m，4H)，2.41(t，J＝7.0Hz，2H)，1.95-1.90(m，2H)，1.80-1.75(m，4H).

(E)-preparation of 3-(5-(3-(morphine quinoline base) propyl group)-2-benzyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl) methyl acrylate (I-7t)

The preparation of reference compound I-7l, I-6l replaces with I-6m with raw material, and piperidines replaces with the morphine quinoline, faint yellow oily thing I-7t, yield 99%

¹H?NMR(400MHz，CDCl ₃)：δ(ppm)：7.80(d，J＝15.9Hz，1H)，7.54(s，1H)，7.42(d，J＝7.2Hz，2H)，7.37-7.28(m，5H)，6.38(d，J＝15.9Hz，1H)，4.09(t，J＝6.7Hz，2H)，3.80(s，2H)，3.79(s，3H)，3.73(s，2H)，3.72(t，J＝4.2Hz，4H)，2.90(t，J＝5.0Hz，2H)，2.84(t，J＝5.0Hz，2H)，2.42-2.35(m，4H)，2.27(t，J＝6.7Hz，2H)，1.93-1.88(m，2H)

¹³C?NMR(100MHz，CDCl ₃)δ168.02，146.86，138.59，137.59，135.11，129.11，128.42，127.25，126.18，125.58，120.65，119.20，114.07，109.31，109.17，66.91，62.42，58.10，54.07，51.45，50.14，49.69，41.06，23.05.

(E)-preparation of 3-(5-(2-(piperidines-1-yl) ethyl)-2-propargyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl) methyl acrylate (I-7u)

The preparation method of reference compound I-7l, faint yellow oily thing I-7u, yield 71%.

¹H?NMR(400MHz，CDCl ₃)δ7.83(d，J＝15.9Hz，1H)，7.58(s，1H)，7.38(dd，J＝8.6，1.3Hz，1H)，7.27(d，J＝8.6Hz，1H)，6.40(d，J＝15.9Hz，1H)，4.13(t，J＝7.5Hz，2H)，3.85(s，2H)，3.80(s，3H)，3.61(d，J＝2.3Hz，2H)，3.01(t，J＝5.6Hz，2H)，2.90(t，J＝5.3Hz，2H)，2.56(t，J＝7.5Hz，2H)，2.47-2.43(m，4H)，2.33(t，J＝2.3Hz，1H)，1.64-1.54(m，4H)，1.47-1.42(m，2H).

¹³C?NMR(100MHz，CDCl ₃)δ168.10，146.84，137.63，134.49，126.02，125.55，120.65，119.10，114.03，109.36，108.70，78.91，73.41，58.45，55.07，51.46，49.33，48.15，46.51，41.24，25.94，24.18，22.98.

(E)-preparation of 3-(5-(2-(piperidines-1-yl) ethyl)-2-allyl group-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl) methyl acrylate (I-7v)

The preparation method of reference compound I-7l, faint yellow oily thing I-7v, yield 65%.

¹H?NMR(400MHz，CDCl ₃)δ7.81(d，J＝15.9Hz，1H)，7.55(s，1H)，7.34(dd，J＝8.6，1.2Hz，1H)，7.22(d，J＝8.5Hz，1H)，6.38(d，J＝15.9Hz，1H)，6.06-5.90(m，1H)，5.38-5.15(m，2H)，4.07(t，J＝7.5Hz，2H)，3.77(s，3H)，3.68(s，2H)，3.27(d，J＝6.5Hz，2H)，2.91-2.80(m，4H)，2.52(t，J＝7.5Hz，2H)，2.43-2.37(m，4H)，1.61-1.51(m，4H)，1.46-1.38(m，2H).

¹³C?NMR(100MHz，CDCl ₃)δ168.06，146.91，137.61，135.50，134.97，126.10，125.45，120.55，119.10，117.98，113.90，109.30，108.94，61.09，58.46，55.04，51.41，50.05，49.48，41.19，25.95，24.19，22.89.

(E)-preparation of 3-(5-(2-(piperidines-1-yl) ethyl)-2-(2-(5-bromo-1H-indoles-1-yl) ethyl)-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl) methyl acrylate (I-7w)

The preparation method of reference compound I-7l, faint yellow oily thing I-7w, yield 58%.

¹H?NMR(400MHz，CDCl ₃)δ7.74(d，J＝15.9Hz，1H)，7.65(d，J＝1.0Hz，1H)，7.46(s，1H)，7.27(dt，J＝8.3，2.1Hz，1H)，7.19-7.16(m，3H)，7.09(d，J＝3.1Hz，1H)，6.33(s，1H)，6.31(d，J＝12.0Hz，1H)，4.22(t，J＝6.8Hz，2H)，4.00(d，J＝7.4Hz，2H)，3.70(s，3H)，3.69(s，2H)，2.90(t，J＝6.8Hz，2H)，2.76-2.68(m，4H)，2.45(d，J＝7.4Hz，2H)，2.37-2.29(m，4H)，1.54-1.44(m，4H)，1.39-1.30(m，2H).

¹³C?NMR(101MHz，CDCl ₃)δ168.12，146.85，137.58，134.95，134.68，130.31，129.38，125.99，125.63，124.29，123.46，120.73，119.04，114.13，112.68，110.81，109.46，108.41，100.96，58.47，57.03，55.11，51.52，50.86，49.88，45.21，41.31，26.00，24.22，22.89.

(E)-preparation of 3-(5-(2-(piperidines-1-yl) ethyl)-2-(2-(phthalic imidine-2-yl) ethyl)-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl) methyl acrylate (I-7x)

The preparation method of reference compound I-7l, faint yellow oily thing I-7x, yield 60%.

¹H?NMR(400MHz，CDCl ₃)δ7.79-7.70(m，3H)，7.60-7.55(m，2H)，7.49(s，1H)，7.30-7.26(m，1H)，7.16(d，J＝8.5Hz，1H)，6.34(d，J＝15.9Hz，1H)，4.01(t，J＝7.2Hz，2H)，3.90(t，J＝6.5Hz，2H)，3.74(s，3H)，3.72(s，2H)，2.98-2.83(m，4H)，2.78-2.71(m，2H)，2.47(t，J＝7.3Hz，2H)，2.41-2.31(m，4H)，1.57-1.48(m，4H)，1.45-1.31(m，2H).

¹³C?NMR(100MHz，CDCl ₃)δ168.26，167.99，146.83，137.44，135.18，133.74，132.10，126.00，125.42，123.06，120.47，119.07，113.90，109.27，108.65，58.40，55.00，54.90，51.38，49.88，49.72，41.18，35.72，25.96，24.20，22.78.

Step 8

(E)-preparation of 3-(5-(2-(piperidines-1-yl) ethyl)-2-benzyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl) vinylformic acid (I-8l)

91mg (0.2mmol) Compound I-7l is dissolved in the 2mL methanol solution, adds the aqueous solution of the potassium hydroxide of 1mL (2mmol) 2M.Behind the back flow reaction 2h,, merge organic phase, saturated common salt water washing, concentrating under reduced pressure behind the anhydrous sodium sulfate drying with the reaction solution ethyl acetate extraction.Crude product is directly used in the next step without purification after the vacuum drying oven drying.

The preparation method of the preparation reference compound I-8l of Compound I-8a～x.

Step 9

(E)-preparation of 3-(5-(2-(piperidines-1-yl) ethyl)-2-benzyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl)-N-((tetrahydrochysene-2H-pyrans-2-yl) oxygen base) acrylamide (I-9l)

Method 1: under the condition of ice bath, 44.5mg (0.1mmol) Compound I-8l is dissolved among the 10mLTHF, adds 0.03mL (0.2mmol) triethylamine and 0.014mL (0.15mmol) Vinyl chloroformate.Azanol (the NH that adds the protection of 23.5mg (0.2mmol) tetrahydropyrans behind the reaction 0.5h again ₂OTHP), reaction solution is moved to room temperature reaction 2h.After reaction finishes, add proper silica gel toward reaction solution in, decompression is spin-dried for afterwards that the sample column chromatography obtains 31mg oily product I-9l, yield 57% on the dry method.

Method 2: with 88mg (0.2mmol) Compound I-8l; 42mg (0.22mmol) 1-ethyl-3-(3-dimethylamine propyl) carbodiimide hydrochloride; 13mg (0.5mmol) N-hydroxyl benzotriazole; the azanol of 48mg (0.4mmol) tetrahydropyrans protection joins in the 25mL round-bottomed flask, adds the 5mL dimethyl formamide again as reaction solvent.Room temperature condition down behind the reaction 24h, adds the water ethyl acetate extraction toward reaction solution, merges organic phase, after decompression is spin-dried on the dry method sample column chromatography obtain 30mg oily product I-9l, yield 28%.

Method 1 in the preparation of the preparation reference compound I-9l of Compound I-9a～x.

Step 10

(E)-preparation of 3-(5-(2-(piperidines-1-yl) ethyl)-2-benzyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl)-N-hydroxyacrylamide dihydrochloride (I-10l)

In 10mL single port pyriform bottle, 15.3mg (0.033mmol) Compound I-9l is dissolved in 1mL exsiccant 1, in 4-dioxy six alkane, stir adding 4M dioxy six heptane hydrochloride solution 0.5mL down, there are a large amount of white precipitates to separate out, continue at room temperature to stir 2h.Reaction finishes, be evaporated to dried, in resistates, add anhydrous diethyl ether 3mL, standing demix behind the high degree of agitation 30min, and drain ether layer, and then add the 3mL anhydrous diethyl ether, drain again after stirring 30min, repeat 3 times after, solid is evaporated to dried, get 8.7mg pink crude product I-10l, yield 64%.

¹H?NMR(400MHz，DMSO)：δ(ppm)：11.48(s，2H)，7.75-7.70(m，3H)，7.66(s，1H)，7.55-7.48(m，4H)，7.42(d，J＝8.6Hz，1H)，6.42(d，J＝15.8Hz，1H)，4.68(t，J＝7.9Hz，2H)，4.60-4.46(m，4H)，4.36-4.29(m，2H)，3.54-3.43(m，2H)，3.34-3.20(m，4H)，2.99-2.89(m，2H)，1.89-1.68(m，6H).

(E)-preparation of 3-(5-(2-(piperidines-1-yl) ethyl)-2-o-cyanobenzyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl)-N-hydroxyacrylamide dihydrochloride (I-10a)

The preparation method of reference compound I-10l, I-9l replaces with I-9a with raw material, gets white-yellowish solid I-10a, yield 74%.

¹H?NMR(400MHz，DMSO)δ11.80(s，1H)，11.41(s，1H)，8.21(s，1H)，8.01(d，J＝7.3Hz，1H)，7.86(t，J＝7.6Hz，1H)，7.78-7.67(m，3H)，7.53(d，J＝15.7Hz，1H)，7.43(d，J＝8.0Hz，1H)，6.43(d，J＝15.6Hz，1H)，4.78-4.61(m，4H)，4.58-4.37(m，2H)，3.88-3.21(m，8H)，3.04-2.88(m，2H)，1.90-1.69(m，6H).

(E)-preparation of 3-(5-(2-(piperidines-1-yl) ethyl)-2-to nitrobenzyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl)-N-hydroxyacrylamide dihydrochloride (I-10b)

The preparation method of reference compound I-10l, I-9l replaces with I-9b with raw material, gets beige solid I-10b, yield 70%.

¹H?NMR(400MHz，DMSO)δ11.68(s，1H)，11.26(s，1H)，10.68(s，1H)，8.36(d，J＝8.3Hz，2H)，8.03(d，J＝7.8Hz，2H)，7.78-7.63(m，2H)，7.51(d，J＝15.4Hz，1H)，7.42(d，J＝8.5Hz，1H)，6.40(d，J＝15.6Hz，1H)，4.70(d，J＝33.1Hz，4H)，4.36(s，2H)，3.59-3.19(m，8H)，3.02-2.84(m，2H)，1.92-1.64(m，6H).

(E)-preparation of 3-(5-(2-(piperidines-1-yl) ethyl)-2-to bromobenzyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl)-N-hydroxyacrylamide dihydrochloride (I-10c)

The preparation method of reference compound I-10l, I-9l replaces with I-9c with raw material, gets yellow solid I-10c, yield 68%.

¹H?NMR(400MHz，DMSO)δ11.79(s，1H)，11.54(s，1H)，7.76-7.67(m，5H)，7.52(d，J＝15.7Hz，1H)，7.41(d，J＝8.4Hz，1H)，6.43(d，J＝15.7Hz，1H)，4.74-4.62(m，2H)，4.62-4.44(m，4H)，4.41-4.19(m，2H)，3.56-3.40(m，2H)，3.34-3.18(m，4H)，3.00-2.87(m，2H)，1.93-1.66(m，6H).

(E)-preparation of 3-(5-(2-(piperidines-1-yl) ethyl)-2-α methylnaphthalene-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl)-N-hydroxyacrylamide dihydrochloride (I-10d)

The preparation method of reference compound I-10l, I-9l replaces with I-9d with raw material, gets white-yellowish solid I-10d, yield 64%.

¹H?NMR(400MHz，DMSO)δ11.45(s，1H)，10.94(s，1H)，8.48-8.33(m，1H)，8.11(d，J＝8.0Hz，1H)，8.09-8.03(m，1H)，7.98(d，J＝6.7Hz，1H)，7.81-7.61(m，5H)，7.51(d，J＝15.2Hz，1H)，7.43(d，J＝8.4Hz，1H)，6.40(d，J＝15.7Hz，1H)，5.15-4.97(m，2H)，4.75-4.61(m，2H)，4.54-4.42(m，2H)，3.94-3.70(m，2H)，3.62-3.23(m，6H)，3.00-2.87(m，2H)，1.95-1.67(m，6H).

(E)-preparation of 3-(5-(2-(piperidines-1-yl) ethyl)-2-β methylnaphthalene-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl)-N-hydroxyacrylamide dihydrochloride (I-10e)

The preparation method of reference compound I-10l, I-9l replaces with I-9e with raw material, gets brown solid I-10e, yield 65%.

¹H?NMR(400MHz，DMSO)δ11.54(s，1H)，11.42(s，1H)，8.23(s，1H)，8.05(d，J＝8.5Hz，1H)，8.03-7.95(m，2H)，7.90(d，J＝8.3Hz，1H)，7.77-7.58(m，4H)，7.50(d，J＝15.5Hz，1H)，7.42(d，J＝8.1Hz，1H)，6.39(d，J＝15.8Hz，1H)，4.83-4.61(m，4H)，4.45-4.34(m，2H)，3.94-3.86(m，2H)，3.77-.21(m，6H)，3.02-2.87(m，2H)，1.95-1.66(m，6H).

(E)-preparation of 3-(5-(2-(piperidines-1-yl) ethyl)-2-to methoxy-benzyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl)-N-hydroxyacrylamide dihydrochloride (I-10f)

The preparation method of reference compound I-10l, I-9l replaces with I-9f with raw material, gets faint yellow solid I-10f, yield 79%.

¹H?NMR(400MHz，DMSO)δ11.56(s，2H)，7.74(d，J＝8.5Hz，1H)，7.68-7.62(m，3H)，7.52(d，J＝15.7Hz，1H)，7.41(d，J＝8.3Hz，1H)，7.04(d，J＝8.3Hz，2H)，6.44(d，J＝15.8Hz，1H)，4.73-4.61(m，2H)，4.58-4.21(m，6H)，3.80(s，3H)，3.56-3.45(m，2H)，3.34-3.19(m，4H)，3.00-2.87(m，2H)，1.92-1.65(m，6H).

(E)-preparation of 3-(5-(2-(piperidines-1-yl) ethyl)-2-p-toluenesulfonyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl)-N-hydroxyacrylamide dihydrochloride (I-10g)

The preparation method of reference compound I-10l, I-9l replaces with I-9g with raw material, gets yellow solid I-10g, yield 63%.

¹H?NMR(400MHz，DMSO)δ11.20(s，1H)，7.76(d，J＝8.0Hz，2H)，7.71(s，1H)，7.65(d，J＝8.5Hz，1H)，7.52(d，J＝15.6Hz，1H)，7.44(d，J＝8.0Hz，2H)，7.37(d，J＝8.5Hz，1H)，6.44(d，J＝15.7Hz，1H)，4.61-4.55(m，2H)，4.27(s，3H)，3.58-3.42(m，4H)，3.23-3.12(m，2H)，3.04-2.83(m，4H)，1.88-1.67(m，6H).

(E)-preparation of 3-(5-(2-(piperidines-1-yl) ethyl)-2-n-propyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl)-N-hydroxyacrylamide dihydrochloride (I-10h)

The preparation method of reference compound I-10l, I-9l replaces with I-9h with raw material, gets yellow solid I-10h, yield 63%.

¹H?NMR(400MHz，DMSO)δ11.47(s，2H)，7.96(s，1H)，7.77(d，J＝8.5Hz，1H)，7.55(d，J＝15.6Hz，1H)，7.43(d，J＝8.3Hz，1H)，6.47(d，J＝15.5Hz，1H)，4.9-4.75(m，2H)，4.55-4.39(m，2H)，4.12-4.02(m，2H)，3.62-3.52(m，2H)，3.33-3.18(m，6H)，3.07-2.94(m，2H)，1.92-1.79(m，2H)，1.58-1.36(m，4H)，1.22-1.14(m，2H)，0.87-0.79(m，3H).

(E)-preparation of 3-(5-(2-(piperidines-1-yl) ethyl)-2-methylsulfonyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl)-N-hydroxyacrylamide dihydrochloride (I-10i)

The preparation method of reference compound I-10l, I-9l replaces with I-9i with raw material, faint yellow solid I-10i, yield 60%.

¹H?NMR(400MHz，DMSO)δ11.34(s，1H)，7.73-7.66(m，2H)，7.54(d，J＝15.6Hz，1H)，7.40(d，J＝8.5Hz，1H)，6.45(d，J＝15.8Hz，1H)，4.69-4.61(m，2H)，4.44(s，2H)，3.66-3.58(m，2H)，3.51(d，J＝11.2Hz，2H)，3.32-3.20(m，2H)，3.07-3.01(m，2H)，2.99(s，2H)，2.96-2.87(m，2H)，1.97-1.69(m，6H).

(E)-preparation of 3-(5-(2-(Pyrrolidine-1-yl) ethyl)-2-benzoyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl)-N-hydroxyacrylamide hydrochloride (I-10j)

The preparation method of reference compound I-10l, I-9l replaces with I-9j with raw material, gets yellow solid I-10j, yield 45%.

¹H?NMR(400MHz，DMSO)δ11.39(s，1H)，10.63(s，1H)，7.85-7.20(m，9H)，6.52-6.31(m，1H)，4.89-4.61(m，2H)，4.59-4.49(m，2H)，4.10-3.65(m，2H)，3.55-3.38(m，4H)，3.08-2.88(m，4H)，2.06-1.94(m，2H)，1.89-1.81(m，2H).

(E)-preparation of 3-(5-(2-(Pyrrolidine-1-yl) ethyl)-2-phenylacetyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl)-N-hydroxyacrylamide hydrochloride (I-10k)

The preparation method of reference compound I-10l, I-9l replaces with I-9k with raw material, gets brown solid I-10k, yield 56%.

¹H?NMR(400MHz，DMSO)δ11.19(s，1H)，10.72(s，1H)，7.99-7.17(m，9H)，6.58-6.36(m，1H)，4.89-4.78(m，1H)，4.77-4.62(m，1H)，4.61-4.47(m，2H)，3.98-3.84(m，2H)，3.59-3.44(m，6H)，3.15-2.80(m，4H)，2.09-1.97(m，2H)，1.96-1.82(m，2H).

(E)-preparation of 3-(5-(3-(piperidines-1-yl) propyl group)-2-benzyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl)-N-hydroxyacrylamide dihydrochloride (I-10m)

The preparation method of reference compound I-10l, I-9l replaces with I-9m with raw material, gets yellow solid I-10m, yield 74%.

¹H?NMR(400MHz，DMSO)δ11.83(s，1H)，10.71(s，1H)，7.80-7.74(m，2H)，7.63(s，1H)，7.59(d，J＝8.6Hz，1H)，7.54-7.46(m，4H)，7.39(d，J＝8.5Hz，1H)，6.43(d，J＝15.8Hz，1H)，4.63-4.47(m，2H)，4.40-4.31(m，2H)，4.29-4.14(m，2H)，3.83-3.45(m，2H)，3.35-3.29(m，2H)，3.09-2.93(m，4H)，2.89-2.70(m，2H)，2.23-2.09(m，2H)，1.85-1.58(m，6H).

(E)-preparation of 3-(5-(2-(Pyrrolidine-1-yl) ethyl)-2-benzyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl)-N-hydroxyacrylamide dihydrochloride (I-10o)

The preparation method of reference compound I-10l, I-9l replaces with I-9o with raw material, gets khaki color solid I-10o, yield 58%.

¹H?NMR(400MHz，DMSO)δ11.71(s，1H)，11.56(s，1H)，7.76-7.68(m，3H)，7.66(s，1H)，7.56-7.46(m，4H)，7.42(d，J＝8.0Hz，1H)，6.42(d，J＝15.9Hz，1H)，4.68-4.45(m，4H)，4.42-4.25(m，2H)，3.83-3.20(m，8H)，3.07-2.86(m，2H)，2.01-1.92(m，2H)，1.90-1.78(m，2H).

(E)-preparation of 3-(5-(2-(morphine quinoline base) ethyl)-2-benzyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl)-N-hydroxyacrylamide dihydrochloride (I-10p)

The preparation method of reference compound I-10l, I-9l replaces with I-9p with raw material, gets faint yellow solid I-10p, yield 45%.

¹H?NMR(400MHz，DMSO)δ12.34-12.19(m，2H)，11.54(s，1H)，7.75-7.70(m，3H)，7.67(s，1H)，7.55-7.48(m，4H)，7.43(d，J＝8.5Hz，1H)，6.42(d，J＝15.8Hz，2H)，4.68(t，J＝7.8Hz，2H)，4.60-4.46(m，2H)，4.39-4.26(m，2H)，4.04-3.94(m，2H)，3.91-3.80(m，2H)，3.54-3.26(m，6H)，3.21-3.10(m，2H).

(E)-preparation of 3-(5-(2-(diethylin) ethyl)-2-benzyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl)-N-hydroxyacrylamide dihydrochloride (I-10q)

The preparation method of reference compound I-10l, I-9l replaces with I-9q with raw material, gets brown solid I-10q, yield 60%.

¹H?NMR(400MHz，DMSO)δ11.70(s，1H)，11.61(s，1H)，7.79-7.69(m，4H)，7.65(s，1H)，7.51(d，J＝16.0Hz，5H)，7.42(d，J＝8.5Hz，1H)，6.43(d，J＝15.7Hz，1H)，4.69(t，J＝7.7Hz，2H)，4.61-4.46(m，2H)，4.40-4.26(m，2H)，3.51-3.10(m，10H)，1.25(t，J＝7.1Hz，6H).

(E)-preparation of 3-(5-(3-(diethylin) propyl group)-2-benzyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl)-N-hydroxyacrylamide dihydrochloride (I-10r)

The preparation method of reference compound I-10l, I-9l replaces with I-9r with raw material, gets powder yellow solid I-10r, yield 86%.

¹H?NMR(400MHz，DMSO)δ11.80(s，1H)，10.71(s，1H)，7.78-7.73(m，2H)，7.66-7.61(m，2H)，7.55-7.47(m，4H)，7.40(d，J＝8.4Hz，1H)，6.43(d，J＝15.8Hz，1H)，4.64-4.21(m，6H)，3.49-3.15(m，4H)，3.04(s，6H)，2.15-2.02(m，2H)，1.18(t，J＝7.2Hz，3H)，1.11(t，J＝7.2Hz，3H).

(E)-preparation of 3-(5-(3-(Pyrrolidine-1-yl) propyl group)-2-benzyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl)-N-hydroxyacrylamide dihydrochloride (I-10s)

The preparation method of reference compound I-10l, I-9l replaces with I-9s with raw material, gets pale brown look solid I-10s, yield 52%.

¹HNMR(400MHz，DMSO)δ11.74(s，1H)，11.14(s，1H)，7.79-7.73(m，2H)，7.63(s，1H)，7.60(d，J＝8.6Hz，1H)，7.55-7.47(m，4H)，7.39(d，J＝8.4Hz，1H)，6.42(d，J＝15.9Hz，1H)，4.63-4.45(m，2H)，4.40-4.16(m，4H)，3.52-3.41(m，2H)，3.35-3.07(m，4H)，3.04-2.84(m，2H)，2.16-2.04(m，2H)，1.96-1.81(m，4H).

(E)-preparation of 3-(5-(3-(morphine quinoline) propyl group)-2-benzyl-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl)-N-hydroxyacrylamide dihydrochloride (I-10t)

The preparation method of reference compound I-10l, I-9l replaces with I-9t with raw material, gets red-brown solid I-10t, yield 76%.

¹H?NMR(400MHz，DMSO)δ11.65(s，1H)，11.39(s，1H)，7.77-7.72(m，1H)，7.64(s，1H)，7.60(d，J＝8.7Hz，1H)，7.55-7.48(m，3H)，7.40(d，J＝8.5Hz，1H)，6.42(d，J＝15.8Hz，1H)，4.60-4.48(m，2H)，4.40-4.17(m，4H)，3.97-3.85(m，4H)，3.84-3.72(m，4H)，3.37-3.28(m，2H)，3.13-3.06(m，2H)，3.05-2.91(m，2H)，2.24-2.09(m，2H).

(E)-preparation of 3-(5-(2-(piperidines-1-yl) ethyl)-2-propargyl-2,3,4, pyridine [4, the 3-b] indoles-8-yl of 5-tetrahydrochysene-1H))-N-hydroxyacrylamide dihydrochloride (I-10u)

The preparation method of reference compound I-10l, I-9l replaces with I-9u with raw material, gets pale brown look solid I-10u, yield 76%.

¹H?NMR(400MHz，MeOD)δ7.67(d，J＝15.3Hz，1H)，7.58(s，1H)，7.49-7.34(m，2H)，6.41(d，J＝15.9Hz，1H)，4.40-4.27(m，2H)，3.92-3.74(m，2H)，3.67-3.57(m，2H)，3.14-3.04(m，2H)，3.01-2.93(m，2H)，2.88-2.67(m，7H)，1.78-1.68(m，4H)，1.62-1.51(m，2H).

(E)-preparation of 3-(5-(2-(piperidines-1-yl) ethyl)-2-allyl group-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl)-N-hydroxyacrylamide dihydrochloride (I-10v)

The preparation method of reference compound I-10l, I-9l replaces with I-9v with raw material, gets pale brown look solid I-10v, yield 65%.

¹H?NMR(400MHz，DMSO)δ7.56-7.49(m，2H)，7.39(d，J＝8.3Hz，1H)，7.28(d，J＝8.1Hz，1H)，6.39(d，J＝15.5Hz，1H)，6.01-5.85(m，1H)，5.27(d，J＝17.1Hz，1H)，5.18(d，J＝10.1Hz，1H)，4.18-4.10(m，2H)，3.64-3.53(m，2H)，3.23(t，J＝12.8Hz，2H)，2.85-2.74(m，4H)，2.56-2.44(m，2H)，2.36-2.28(m，4H)，1.50-1.39(m，4H)，1.38-1.30(m，2H).

(E)-preparation of 3-(5-(2-(piperidines-1-yl) ethyl)-2-(2-(5-bromo-1H-indoles-1-yl) ethyl)-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl)-N-hydroxyacrylamide dihydrochloride (I-10w)

The preparation method of reference compound I-10l, I-9l replaces with I-9w with raw material, gets pale brown look solid I-10w, yield 70%.

¹H?NMR(400MHz，DMSO)δ11.65(s，1H)，11.39(s，1H)，7.74(d，J＝15.9Hz，1H)，7.65(d，J＝1.0Hz，1H)，7.46(s，1H)，7.27(dt，J＝8.3，2.1Hz，1H)，7.19-7.16(m，3H)，7.09(d，J＝3.1Hz，1H)，6.33(s，1H)，6.31(d，J＝12.0Hz，1H)，4.22(t，J＝6.8Hz，2H)，4.00(d，J＝7.4Hz，2H)，3.69(s，2H)，2.90(t，J＝6.8Hz，2H)，2.76-2.68(m，4H)，2.45(d，J＝7.4Hz，2H)，2.37-2.29(m，4H)，1.54-1.44(m，4H)，1.39-1.30(m，2H).

(E)-preparation of 3-(5-(2-(piperidines-1-yl) ethyl)-2-(2-(phthalic imidine-2-yl) ethyl)-2,3,4,5-tetrahydrochysene-1H-pyridine [4,3-b] indoles-8-yl)-N-hydroxyacrylamide dihydrochloride (I-10x)

The preparation method of reference compound I-10l, I-9l replaces with I-9x with raw material, gets pale brown look solid I-10x, yield 75%.

¹H?NMR(400MHz，DMSO)δ11.65(s，1H)，11.39(s，1H)，7.79-7.70(m，3H)，7.60-7.55(m，2H)，7.49(s，1H)，7.30-7.26(m，1H)，7.16(d，J＝8.5Hz，1H)，6.34(d，J＝15.9Hz，1H)，4.01(t，J＝7.2Hz，2H)，3.90(t，J＝6.5Hz，2H)，3.72(s，2H)，2.98-2.83(m，4H)，2.78-2.71(m，2H)，2.47(t，J＝7.3Hz，2H)，2.41-2.31(m，4H)，1.57-1.48(m，4H)，1.45-1.31(m，2H).

Embodiment 2 compounds of the present invention are to the inhibited proliferation of various tumor cell strains:

1. experiment material:

Cell strain and culture condition: human breast cancer cell strain (MCF-7), human colon cancer cell strain (HCT116), human lung carcinoma cell line (A549), human cervical carcinoma cell strain (HeLa) and human leukemia cell line (K562), above cell strain are all bought from cell resource center of Shanghai Sheng Ke institute of the Chinese Academy of Sciences.MCF-7 is incubated at and contains 10% foetal calf serum (FBS, fetal bovine serum) modified form perfect medium (DMEM, Dulbecco ' s Modified Eagle Medium) in the substratum, A549 is incubated in the DMEM that contains 10%FBS: F12 (1: the 1) substratum, HCT116, HeLa and K562 are incubated in the RPMI1640 substratum that contains 10%FBS, each substratum is all available from GIBCO, and to contain final concentration be the penicillin of 100U/mL and the Streptomycin sulphate of 100U/mL.

2. experimental technique:

(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method detects the inhibited proliferation of each compound to various tumor cell strains to adopt tetrazolium bromide 3-.Collect the logarithmic phase cell, be inoculated in 96 well culture plates with 2 * 104/mL, every hole adds 100 μ L, and in 5% carbonic acid gas (CO ₂), hatch 24h in 37 ℃ of constant incubators after, the compound that adds different concns, 3 multiple holes are established in every hole, and establish blank group (substratum group), negative control group (no medicine group) and positive controls (SAHA group), after hatching 48 hours jointly, every hole adds 20 μ L MTT solution (5mg/mL), continues to cultivate 4h.Stop cultivating, inhale and go nutrient solution in the hole, every hole to add 100 μ L methyl-sulphoxides (DMSO), put low-speed oscillation 10min on the shaking table, crystallisate is fully dissolved.Measure the light absorption value in each hole at enzyme-linked immunosorbent assay instrument OD492/630nm place.With the negative control group is reference, calculates the propagation inhibition IC of different concns compound to tumor cell line ₅₀Value.The calculation formula of cell inhibitory rate is:

Obtain IC with the Bliss method ₅₀

3. experimental result

Compound effects was measured IC respectively after 48 hours among the present invention ₅₀Value.Most compounds obviously are better than positive control medicine million woods letter SAHA (popular name: vorinostat) or suitable with it to the restraining effect of 5 kinds of tumour cells.Compound I-10b wherein, I-10e, I-10g, I-10h, I-10l is for the IC of various tumour cells ₅₀Value obviously is better than SAHA, is example with Compound I-10h, and it is that 30 times of SAHA reach 0.152 μ M to the HCT116 cell activity, reaches about 2～10 times of SAHA for the activity of other tumour cell.They to the vitro inhibition exercising result of tumour cell referring to table 2.

48 hours inhibition activity of table 2 compound effects for tumour cell

Embodiment 3 compounds of the present invention are to the enzyme inhibition activity of external HDACs and HDAC1

Adopt histon deacetylase (HDAC) (HDACs) to suppress active detection kit-histon deacetylase (HDAC) fluorescence detection reagent kit (HDAC Fluorimetric Assay/Drug Discovery Kit; AK-500; Biomol) and histone deacetylase 1 (HDAC1) suppress active detection kit histone deacetylase 1 fluorescence detection reagent kit (HDAC1 Fluorimetric Assay/Drug Kit; AK-511; Biomol), with fluorogenic substrate (Fluror de lys ^TM) be substrate, detect among the present invention compound respectively to HDACs and HDAC1 half-inhibition concentration (IC ₅₀).All operations all provides operational guidance to carry out according to test kit, and the concrete operations step is as follows: configuration waits to try the compound mother liquor, and carries out the gradient concentration dilution.HDACs and HDAC1 assaying reaction are as follows: respectively Hela nucleus extract and HDAC1 enzyme are added in the 96 hole fluorescent plates that test kit provides, and add different concns wait to try compound and Fluror de lys ^TMCarry out hybrid reaction, 3 multiple holes are established in every hole, hatch 45min under the room temperature, add extension reaction solution termination reaction, and adopt the multi-functional microplate reader of high-performance (BioTek, USA) fluorescence intensity (Ex=380nm, Em=480nm).

Adopt Prism Graph pad 5.0 to carry out non-alignment curve match, and calculate IC ₅₀Value, each compound is to the half-inhibition concentration IC of HDACs and HDAC1 ₅₀Value sees Table 3.

Table 3 compound is to the inhibition activity of HDACs and HDAC 1

Wherein nd represents compound is not carried out the test of respective item.

Test result shows, Compound I-10b wherein, and I-10h suppresses IC to HDACs ₅₀Value is better than SAHA, Compound I-10b, and I-10c, I-10g, I-10h, I-10l, I-10o suppresses IC to HDAC1 ₅₀Value is better than or approaches SAHA, demonstrates the restraining effect of good HDACs and HDAC1, has the potentiality that are developed to antitumor drug candidate.

Compound is induced the change (with reference to Taghibiglou, reported method such as C., Nat.Med.2009,15,1399) of neuroblastoma cell SHSY5Y form among embodiment 4 the present invention

To be in logarithmic phase cell (2 * 10 ⁵) be inoculated in 24 orifice plates, add different concns I-10l, control group adds isopyknic RPMI RPMI-1640.Handle centrifugally after 48 hours, remove upper strata RPMI 1640 substratum, wash 2 times with damping fluid (PBS), centrifugal go supernatant (1500rpm, 5min).Add stationary liquid (methyl alcohol: acetate=3: 1), 4 ℃ of fixing 10min.Adopt nucleus dyeing Hoechst 33258 (5 μ L/ml) dyeing 5min, observe under the 356nm burst of ultraviolel, fluorescent microscope and take pictures.

Different concns I-10l sees accompanying drawing 1 to the influence of neuroblastoma SHSY5Y cellular form, and the concentration that I-10l adopts in the accompanying drawing is respectively 0.1 μ M, 0.5 μ M and 1 μ M.

The result shows that the SHSY5Y cell is compared with control group after acting on 48 hours, cell presents that nuclear chromatin concentrates, nuclear fragmentation and typical apoptosis feature (Fig. 1) such as apoptotic body occurs, and with the raising of concentration, the apoptosis feature is obvious all the more, presents dose-dependently; And control group does not have morphological change, and institute fluoresces evenly (Fig. 1), prompting I-10l energy inducing cell generation apoptosis, and this may be one of its mechanism of action.

Compound is to the influence of the cell cycle of neuroblastoma cell line SHSY5Y (with reference to Atadja, reported method such as P., Cancer Research 2004,64,689) among embodiment 5 the present invention

With neuroblastoma SHSY5Y cell (2 * 10 ⁵) be inoculated in six orifice plates, after the I-10l effect, collecting cell also cleans the back with PBS and adopts 70% ethanol fixedly to spend the night in 4 ℃.Discard ethanol after fixing, and with the PBS cleaning, be suspended in 500 μ L iodate third pyrroles (PI) and the ribonuclease A (RNAseA), in dark, hatch 30min for 4 ℃, with flow cytometer (FACS) test sample, Cell Quest software (Becton) and Modifit software obtain and analytical data.

Different concns I-10l sees Fig. 2 and Fig. 3 to the data that influence and the concentration of the cell cycle of SHSY5Y is 1 μ M.

The result shows that I-10l can block cell the phase in G1, and S phase and G2 phase cell all have minimizing in various degree; And the concentration with I-10l strengthens, and G1 phase retardation is obvious all the more.

Compound is to the apoptosis-induced effect of neuroblastoma cell SHSY5Y (with reference to Atadja, reported method such as P., Cancer Research 2004,64,689) among embodiment 6 the present invention

Cell (2 * 10 ⁵) be inoculated in six orifice plates, after the I-10l effect, collecting cell and clean with PBS after, (Annexin V-FITC/PI Promega) detects to adopt the apoptosis test regent box.Provide guide to operate according to the reagent merchant, as follows: as with the cell after the I-10l effect, to be resuspended in Annexin V-FITC, to add 5 μ l fluorescent mark phospholipids incorporate albumen (AnnexinV-FITC), mixings gently in conjunction with in the liquid (1 *); Room temperature (20-25 ℃) lucifuge is hatched 10min, and the centrifugal 5min of 1000g abandons supernatant, adds 195 μ l Annexin V-FITC in conjunction with liquid (1 *) re-suspended cell gently, adds 10 μ l propidium iodide staining fluids (PI), mixing gently, and the ice bath lucifuge is placed 10min; With FACS flow cytometer test sample, Cell Quest software (Becton) and Modifit software obtain and analytical data.

Different concns I-10l sees Fig. 4 and Fig. 5 to the data that influence and the concentration of the cell cycle of neuroblastoma SHSY5Y is 1 μ M.

The result shows that I-10l obviously inducing cell produces apoptosis, and 1 μ M I-10l can induce and produce apoptosis rate near 50%, and demonstrating I-10l can remarkable apoptosis-induced generation, Zong and apoptosis rate increase with the increase of dosage, also demonstrate the regular hour dependency.

Compound is to the tumor inhibition effect of transplanted tumor animal model among embodiment 7 the present invention

With the tumor tissues of well-grown Lewis lung cancer tumor-bearing mice, shred and grind after 80 mesh filter screens filter, be prepared into 5 * 10 ⁶The single cell suspension of cell/mL, the right side armpit that is inoculated into mouse (C57BL/6 mouse, male, SPF level, 4～6 ages in week) is subcutaneous, and every 0.2mL makes the modeling of transplanted tumor animal model, sets up the operation group to give physiological saline separately; Be divided into 5 groups at random behind the inoculation 24h, be respectively positive SAHA group (50mg/kg), negative control group, administration high dose group (50mg/kg), middle dosage group (25mg/kg) and low dose group (12.5mg/kg), after treating a week, it is 100mm that the model group tumour is grown to volume ³Intraperitoneal injection, operation group and negative control group all give isopyknic physiological saline.

Weigh every day after the administration, and observe animal model physiological status and sign.Drug withdrawal after 8 days is weighed in the 24h, and puts to death animal and claim knurl heavy, calculates that respectively to organize average knurl heavy, calculates inhibition rate of tumor growth and carries out the T check by following formula.

1. experimental result:

A. animal is observed

The equal outward appearance of each treated animal is normal after the administration, and fur is glossy, and animal appearance health, active situation are good, and the water of ingesting is normal, no toxic side effects performance.

B. body weight change

Each treated animal body weight all has in various degree to be increased, and it is 4 as follows that statistics sees Table:

Animal model body weight result of variations in table 4 body

The result shows, the operation group is the normal control group, injecting normal saline only, body weight has certain increase, and other tumor model group body weight also have increase, but not statistically significant, but with the increase of dosage, the weight increase amount reduces, and the characteristic of this type of epigenetics medicine high-efficiency low-toxicity is described, with the bibliographical information of positive drug SAHA is consistent, illustrates that also I-10l has the excellent development prospect.

C. tumor control rate calculates

Animal model tumor control rate result in table 5 body

Interpretation of result: from the tumor control rate result as seen, the low dosage tumor control rate has been higher than positive drug SAHA, and each administration group more all has the significance meaning with negative group, and strengthens with dosage, and tumor control rate obviously increases.

Claims

1. the γ carboline compound with following structural formula and pharmaceutically acceptable salt and comprise the pharmaceutical composition of this compound:

Wherein:

L ₁Expression C ₁～C ₄Alkylidene group;

R ², R ³Be C ₁-C ₆Alkyl, C ₁-C ₆The assorted alkyl that contains N, S or O atom, each other can be identical or inequality, or each other in encircling into key.

2. synthetic compound intermediate as claim 1 γ carboline compound, its structural formula such as following:

3. pharmaceutical composition wherein contains the compound that requires just like right 1 and acceptable carrier pharmaceutically.

4. a compound as claimed in claim 1 is used to prepare NSC 630176 or antitumor, leukemic medicine.

5. the preparation method of compound as claimed in claim 1 or 2 is characterized in that realizing as follows:

Wherein X is a halogen; R ¹, R ², R ³, L ₁According to claim 1; R ⁴As described in claim 2; Compound I-7 structural formula is with shown in the intermediate in the claim 2, and Compound I-10 structural formula is shown in claim 1.

6. method as claimed in claim 5 is characterized in that step 2), 3), 6), 7), 8), 9) and 10) organic solvent described in the step is tetrahydrofuran (THF), methylene dichloride, acetonitrile, methyl alcohol, ethanol, dimethyl formamide, toluene and N-Methyl pyrrolidone.

7. method as claimed in claim 5 is characterized in that step 2), 3), 6), 7) and 8) described in alkali or acid binding agent be triethylamine, N, N-diisopropylethylamine, pyridine, salt of wormwood, sodium bicarbonate, sodium hydroxide or yellow soda ash.

8. method as claimed in claim 5 is characterized in that step 3), 9) be under protection of inert gas, to carry out.

9. method as claimed in claim 5 is characterized in that the quaternary ammonium salt described in the step 4) is Tetrabutyl amonium bromide, 4-butyl ammonium hydrogen sulfate or benzyltriethylammoinium chloride.