CN110372680A

CN110372680A - A kind of novel protein inhibitor and its preparation method and application

Info

Publication number: CN110372680A
Application number: CN201810327536.2A
Authority: CN
Inventors: 曹文杰; 程鹏
Original assignee: Changzhou Long Run Medical Technology Co Ltd
Current assignee: Changzhou Long Run Medical Technology Co Ltd
Priority date: 2018-04-12
Filing date: 2018-04-12
Publication date: 2019-10-25

Abstract

The present invention provides a kind of novel protein inhibitor and its preparation method and application.Specifically, the present invention provides a kind of such as following formula I compound represented；Wherein, the definition of each group is as noted in the discussion.The compound of the present invention has good Bcl-2 family protein inhibitory activity, can be used for preparing a series of drug of the relevant disease for the treatment of Bcl-2 family protein activity.

Description

A kind of novel protein inhibitor and its preparation method and application

Technical field

The invention belongs to drug fields, in particular it relates to a kind of quinoline azoles as Bcl-2 family protein inhibitor Quinoline class compound, and its preparation and application.

Background technique

Apoptosis is the process of a height control, it guides cell to carry out self degradation to eliminate the thin of malfunction Born of the same parents.During tumour is continued to develop to precarious position, it must suppress the effect of Apoptosis.It is special in tumour cell Property active cell apoptosis provide a kind of scheme of novel neoplasm targeted therapy.The biological targets of product are researched and developed in this project Aiming at the major control point in apoptosis process.

Apoptosis be divided in external two main paths.Inherent approach is withered by the rush of control mitochondrial outer membrane integrality Die and two class Bcl-2 albumen of anti-apoptotic between interaction and regulate and control.Rush apoptosis member (Bax, Bak, Bad, Bik, Bid, Bim, Hrk, Bmf, Noxa and Puma) inducing mitochondrial membrane permeability increase, discharge cytochrome c, activate Guang door Protease is to start apoptotic process.Anti-apoptotic member (Bcl-2, Bcl-xL, Bcl-w, Mcl-1, and Bcl2-A1) is by combining Promote apoptosis member thus the generation for preventing Apoptosis with being isolated.Cancer cell often over-express the Bcl-2 family of anti-apoptotic at Member inhibits Apoptosis, improves cells survival rate, is formed and is resisted in chemotherapy.Inhibit the effect of anti-apoptotic Bcl-2 family member It has been studied the effective means for being proved to be and inducing Apoptosis in cancer cell again, has been confirmed to be the important of clinical treatment intervention Biological targets.It includes acute lymphoblastic leukemia (ALL) that the cancer of anti-apoptotic Bcl-2 albumen is over-expressed in cell, slowly Property lymphocytic leukemia (CLL), Huppert's disease, mankind's follicular lymphoma, Small Cell Lung Cancer (SCLC), uterine cancer, Oophoroma, bladder cancer, prostate cancer, cancer of pancreas, gastric cancer, colorectal cancer and breast cancer etc..

Therefore, protein-protein interaction (PPI) inhibitor for developing apoptosis of tumor cells approach has become anti-swell The forward position focus of tumor medicine research.

Summary of the invention

The object of the present invention is to provide a kind of novel targeted Bcl-2 family protein inhibitors.

The first aspect of the present invention provides a kind of such as following formula I compound represented or its pharmaceutically acceptable salt:

Wherein,

Ring A is group selected from the group below:

R¹And R²It is each independently selected from: H, halogen, NO₂、CN、CF₃SO₂Or CF₃；

R³And R⁴It is each independently selected from: H, substituted or unsubstituted C1-C8 alkyl, C (O) OR⁵、CONR⁶R⁷、C(O)R⁸, replace Or unsubstituted 5~8 yuan of aryl, substituted or unsubstituted 5~8 unit's heteroaryl, it is substituted or unsubstituted 3~12 yuan saturation or not Saturated heterocyclic or carbocyclic ring；Wherein, the heteroaryl includes 1-3 hetero atoms selected from the group below: N, O or S；The heterocycle packet Containing 1-3 hetero atoms selected from the group below: N, O or S；

R⁵、R⁶、R⁷And R⁸It is each independently selected from: H, substituted or unsubstituted C1-C8 alkyl, substituted or unsubstituted C1-C8 alkane Oxygroup, substituted or unsubstituted C1-C6 alkylidene-amido, substituted or unsubstituted C1-C6 alkylidene-hydroxyl, substitution do not take 5~8 yuan of aryl in generation or substituted or unsubstituted 5~8 unit's heteroaryl；Substituted or unsubstituted 3~12 yuan of saturated heterocyclics or carbon Ring；Wherein, the heteroaryl includes at least one hetero atom selected from the group below: N, O or S, and the heterocycle includes at least one A hetero atom selected from the group below: N, O or S；

Above-mentioned any " substitution " refers to that one or more hydrogen atoms on group are replaced by substituent group selected from the group below: halogen, OH, NH₂, CN, unsubstituted or halogenated C1-C8 alkyl, C1-C8 alkoxyl, unsubstituted or halogenated C2-C6 alkenyl, unsubstituted or halogen It is the C2-C6 alkynyl in generation, unsubstituted or halogenated C2-C6 acyl group, unsubstituted or halogenated 5~8 yuan of aryl, unsubstituted or halogenated 5~8 unit's heteroaryls, unsubstituted or halogenated 3~12 yuan of saturated heterocyclics or carbocyclic ring；Wherein, the heteroaryl includes 1-3 choosing From the hetero atom of the following group: N, O or S, the heterocycle include 1~3 hetero atom selected from the group below: N, O or S.

In another preferred example, ring A is

In another preferred example, work as R¹It is selected from: halogen, NO₂、CN、CF₃SO₂Or CF₃When, R²For H.

In another preferred example, work as R¹When for H, R²It is selected from: halogen, NO₂、CN、CF₃SO₂Or CF₃。

In another preferred example, work as R¹For NO₂Or CF₃SO₂When, R²For H.

In another preferred example, work as R²For NO₂Or CF₃SO₂When, R¹For H.

In another preferred example, R³And R⁴It is each independently selected from: H, substituted or unsubstituted C1-C8 alkyl.

In another preferred example, R³Or R⁴In one of them be H and another is the C1-C8 alkyl replaced.

In another preferred example, A, R¹、R²、R³Or R⁴For group corresponding in particular compound described in embodiment.

In another preferred example, the compound that the compound of formula I is as follows:

The second aspect of the present invention provides a kind of preparation method of compound of formula I as described in the first aspect of the invention, Comprising steps of

(a) it in atent solvent, is reacted with -13 compound of Formulas I and -14 compound of Formulas I, obtains compound of formula I；

In the above formulas, each group is as defined above.

In another preferred example, in (a) step, the atent solvent is selected from the group: methylene chloride, methanol, second Alcohol, isopropanol, n-butanol, the tert-butyl alcohol, isobutanol, dioxane, THF, DMF, DMSO, NMP, or combinations thereof.

In another preferred example, in (a) step, the reaction carries out in the presence of a base.

In another preferred example, in (a) step, the alkali is selected from the group: triethylamine, diisopropylamine, two Wopropyl ethyl amine, Na₂CO₃、K₂CO₃、Cs₂CO₃, LiHMDS, NaHMDS, KHMDS, sodium tert-butoxide, potassium tert-butoxide, or combinations thereof.

In another preferred example, the preparation method is further comprising the steps of:

(b) it in atent solvent, is reacted with -12 compound of Formulas I and -5 compound of Formulas I, obtains -13 compound of Formulas I；

In the above formulas, each group is as defined above.

In another preferred example, in (b) step, the atent solvent is selected from the group: toluene, dioxane, THF, DMF, or combinations thereof.

In another preferred example, in (b) step, the reaction carries out in the presence of palladium catalyst.

In another preferred example, in (b) step, the palladium catalyst is selected from the group: Pd (PPh₃)₄、Pd₂ (dba)₃、Pd(dba)₂、Pd(OAc)₂、Pd(PPh₃)₂Cl₂、Pd(dppe)Cl₂、Pd(dppf)Cl₂、Pd(dppf)Cl₂· CH₂Cl₂, or combinations thereof.

In another preferred example, in (b) step, the reaction carries out in the presence of mantoquita.

In another preferred example, in (b) step, the mantoquita is selected from the group: CuI, Cu, CuCl, Cu₂O、 CuO、Cu(OAc)₂、CuSO₄·5H₂O、Cu(acac)₂、CuCl₂, CuSCN, or combinations thereof.

In another preferred example, in (b) step, the reaction carries out in the presence of a base.

In another preferred example, in (b) step, the alkali is selected from the group: K₂CO₃、K₃PO₄、Cs₂CO₃Or its Combination.

The third aspect of the present invention provides a kind of purposes of compound as described in the first aspect of the invention, is used for:

(a) drug of preparation treatment disease relevant to Bcl-2 family protein activity or expression quantity；

(b) the targeted inhibition agent of Bcl-2 family protein is prepared；

(c) inhibit to external non-therapeutic the activity of Bcl-2 family protein；

(d) inhibit tumor cell proliferation to external non-therapeutic；And/or

(e) treatment disease relevant to Bcl-2 family protein activity or expression quantity.

In another preferred example, the Bcl-2 family protein is selected from the group: Bcl-2, Bcl-xL, Bcl-w, Mcl-1, Bcl2-A1, or combinations thereof.

In another preferred example, the tumour cell is leukemia cell line；Preferably lymphocytic leukemia is thin Born of the same parents' strain；It is more preferably acute lymphoblastic leukemia cell strain MV-4-11 cell.

The fourth aspect of the present invention, provides a kind of pharmaceutical composition, and the pharmaceutical composition includes: (i) effective quantity Compound of formula I or its pharmaceutically acceptable salt；(ii) pharmaceutically acceptable carrier.

In another preferred example, the effective quantity refers to therapeutically effective amount or inhibits effective quantity, preferably 0.01~ 99.99%.

In another preferred example, the pharmaceutical composition is used to inhibit the activity of Bcl-2 family protein.

In another preferred example, the pharmaceutical composition is for treating and Bcl-2 family protein activity or expression quantity phase The disease of pass.

The fifth aspect of the present invention provides a kind of active method of inhibition Bcl-2 family protein, comprising steps of to suppression Object application processed inhibits a effective amount of compound of formula I as described in the first aspect of the invention or its pharmaceutically acceptable salt, or Inhibit a effective amount of pharmaceutical composition as described in the fourth aspect of the present invention to inhibiting object to apply.

In another preferred example, the inhibition is the inhibition of external non-therapeutic.

In another preferred example, when to inhibiting, object applies a effective amount of above-mentioned compound of formula I of inhibition or it pharmaceutically may be used When the salt of receiving, the inhibition effective quantity is 0.001-500nmol/L, preferably 0.01-200nmol/L.

The sixth aspect of the present invention provides a kind of method of extracorporeal suppression tumor cell, which comprises to inhibition Object application inhibits a effective amount of compound of formula I or as described in the fourth aspect of the present invention as described in the first aspect of the invention Pharmaceutical composition.

It should be understood that above-mentioned each technical characteristic of the invention and having in below (eg embodiment) within the scope of the present invention It can be combined with each other between each technical characteristic of body description, to form a new or preferred technical solution.As space is limited, exist This no longer tires out one by one states.

Specific embodiment

The present inventor's in-depth study by long-term is prepared for a kind of compound with structure shown in Formulas I, and finds It is with Bcl-2 family protein inhibitory activity.And the compound is at extremely low concentration (can be down to≤100nmol/L), i.e., Inhibiting effect is generated to a series of Bcl-2 family proteins, inhibitory activity is quite excellent, thus can be used for treating and Bcl-2 family Race's protein active or the relevant disease of expression quantity such as tumour.Based on above-mentioned discovery, inventor completes the present invention.

Term

As used herein, term " C1-C6 alkyl " refers to the linear or branched alkyl group with 1~6 carbon atom, such as methyl, second Base, propyl, isopropyl, butyl, isobutyl group, sec-butyl, tert-butyl, or similar group.Term " C1-C8 alkyl " refers to 1~8 The linear or branched alkyl group of a carbon atom, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl group, sec-butyl, tert-butyl, Or similar group.Term " C1-C8 alkoxyl " refers to C1-C8 straight or branched alkoxyl, refers to the alkane containing 1-8 carbon atom Oxygroup, including but not limited to methoxyl group, ethyoxyl, positive propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, Tert-butoxy, amoxy, hexyloxy, oxygroup in heptan or octyloxy.

As used herein, term " C2-C6 acyl group " finger-type such as " linear or branched alkyl group-carbonyl with 1~6 carbon atom The substituent group of base " structure, such as acetyl group, propiono, bytyry, or similar group.

Term " C1-C6 alkylidene " refers to that C1~C6 alkyl as described above loses the base that a hydrogen atom is formed later Group, such as-CH₂-、-CH₂-CH₂, or similar group.Term " C2-C6 alkenyl " refer to 2~6 carbon atoms and include to The linear chain or branched chain alkyl of a few double bond.Term " C2-C6 alkynyl " refers to 2~6 carbon atoms and includes at least one The linear chain or branched chain alkyl of three keys.

Term " C6-C10 arlydene " refers to that the aryl with 6-10 carbon atom loses the group of hydrogen atom formation, packet Include monocycle or two ring arlydene, such as phenylene, naphthylene, or similar group.

Term " 6 yuan of aryl " refers to phenyl.

Term " 5~8 yuan of aryl " refers to the substituent group of the carbon unsaturation system of 5-8 member ring, such as phenyl, or similar group.

Term " 5 yuan~8 unit's heteroaryl ", which refers in the ring system with 5-8 member, has one or more selected from the miscellaneous of O, S, N or P The unsaturation ring system substituent of atom, such as pyridyl group, thienyl, or similar group.

Term " 3~12 yuan of saturated carbon rings " refers to the saturated carbon ring with 3-12 carbon atom, such as cyclohexyl, or similar base Group.Term " 3~12 yuan of unsaturated carbocyclics " refers to the unsaturated carbocyclic with 3-12 carbon atom, such as cyclohexenyl group or similar Group.

Term " 3~12 yuan of saturated heterocyclics ", which refers in the ring system with 3-12 member, has one or more selected from O, S, N or P Heteroatomic saturation ring system substituent, such as piperidyl, THP trtrahydropyranyl, morphine forest base, or similar group." 3~12 yuan are not or not term Saturated heterocyclic " refers to, and there are one or more heteroatomic unsaturated ring systems selected from O, S, N or P to take in the ring system with 3-12 member Dai Ji, such as pyrrole radicals, or similar group.

Term " halogen " refers to F, Cl, Br and I.

In the present invention, term " containing ", "comprising" or " comprising " indicate that various composition can be applied to of the invention mix together It closes in object or composition.Therefore, term " mainly by ... form " and " consist of " were included in term " containing ".

In the present invention, term " pharmaceutically acceptable " ingredient refers to suitable for people and/or animal and without excessive bad pair It reacts (such as toxicity, stimulation and allergy), that is, has the substance of reasonable benefit/risk ratio.

In the present invention, amount or table that term " effective quantity " refers to therapeutic agent treatment, alleviates or prevent target disease or situation Reveal the detectable amount for treating or preventing effect.The figure of the object is depended on for the accurate effective quantity of certain an object and is good for The combination of therapeutic agent and/or therapeutic agent that health situation, the property and degree of illness and selection are given.Therefore, standard is preassigned True effective quantity is useless.However, can determine the effective quantity with routine experiment for the situation that Mr. Yu gives, face Bed doctor can judge.

Herein, except place is illustrated, term " substitution " refers to that one or more hydrogen atoms on group are selected from down The substituent group of group replaces: halogen, unsubstituted or halogenated C1-C6 alkyl, unsubstituted or halogenated C2-C6 acyl group, it is unsubstituted or Halogenated C1-C6 alkyl-hydroxyl, OH, NH₂, CN, unsubstituted or halogenated C1-C8 alkyl, C1-C8 alkoxyl, unsubstituted or halogen It is the C2-C6 alkenyl in generation, unsubstituted or halogenated C2-C6 alkynyl, unsubstituted or halogenated 5~8 yuan of aryl, unsubstituted or halogenated 5~8 unit's heteroaryls, unsubstituted or halogenated 3~12 yuan of saturated heterocyclics or carbocyclic ring；Wherein, the heteroaryl includes 1-3 choosing From the hetero atom of the following group: N, O or S, the heterocycle include 1~3 hetero atom selected from the group below: N, O or S.

Unless stated otherwise, in the present invention, the compound occurred is intended to including all possible optical isomer, Such as the compound of single chiral or the mixture (i.e. racemic modification) of various different chipal compounds.All chemical combination of the invention Among object, each asymmetric carbon atom can be optionally the mixture of R configuration or S configuration or R configuration and S configuration.

As used herein, term " the compounds of this invention " refers to Formulas I compound represented.The term further includes and Formulas I chemical combination Various crystalline forms, pharmaceutically acceptable salt, hydrate or the solvate of object.

As used herein, term " pharmaceutically acceptable salt " refers to that the compounds of this invention and acid or alkali are formed by suitable use Make the salt of drug.Pharmaceutically acceptable salt includes inorganic salts and organic salt.A kind of preferred salt is the compounds of this invention and acid The salt of formation.The acid for suitably forming salt includes but is not limited to: hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, phosphoric acid etc. are inorganic Acid, formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, Picric acid, methanesulfonic acid, benzene methanesulfonic acid, the organic acids such as benzene sulfonic acid；And the acidic amino acids such as aspartic acid, glutamic acid.

Compound of formula I

Wherein,

Ring A is group selected from the group below:

In another preferred example, ring A is

In another preferred example, work as R¹For NO₂Or CF₃SO₂When, R²For H.

In another preferred example, work as R²For NO₂Or CF₃SO₂When, R¹For H.

The preparation method of compound of formula I

The present invention provides a kind of the method for preparing compound of formula I, the described method comprises the following steps:

In the above formulas, each group is as defined above.

In another preferred example, the step carries out in the presence of a base.

In another preferred example, the alkali is selected from the group: triethylamine, diisopropylamine, diisopropylethylamine, Na₂CO₃、 K₂CO₃、Cs₂CO₃, LiHMDS, NaHMDS, KHMDS, sodium tert-butoxide, potassium tert-butoxide, or combinations thereof.

In the above formulas, each group is as defined above.

In another preferred example, the step carries out in the presence of palladium catalyst.

In another preferred example, the palladium catalyst is selected from the group: Pd (PPh₃)₄、Pd₂(dba)₃、Pd(dba)₂、Pd (OAc)₂、Pd(PPh₃)₂Cl₂、Pd(dppe)Cl₂、Pd(dppf)Cl₂、Pd(dppf)Cl₂·CH₂Cl₂, or combinations thereof.

In another preferred example, the step carries out in the presence of mantoquita.

In another preferred example, the mantoquita is selected from the group: CuI, Cu, CuCl, Cu₂O、CuO、Cu(OAc)₂、 CuSO₄·5H₂O、Cu(acac)₂、CuCl₂, CuSCN, or combinations thereof.

In another preferred example, the step carries out in the presence of a base.

In another preferred example, the alkali is selected from the group: K₂CO₃、K₃PO₄、Cs₂CO₃, or combinations thereof.

(c) it in atent solvent, is reacted with -10 compound of Formulas I and -11 compound of Formulas I, obtains -12 compound of Formulas I；

In the above formulas, each group is as defined above.

In another preferred example, in (c) step, the atent solvent is selected from the group: toluene, dioxane, THF、DMF、H₂O, or combinations thereof.

In another preferred example, the step carries out in the presence of ligand.

In another preferred example, the ligand is monodentate phosphine ligand or bidentate phosphine ligands；More preferably, ligand choosing From the following group: triphenylphosphine, trimethylphenyl phosphine, tricyclohexyl phosphine, tri-tert-butylphosphine, X-Phos, S-Phos, dinaphthalene diphenylphosphine, 1,1'- bis- (diphenylphosphine) ferrocene, 1,2- bis- (diphenylphosphino) ethane, Xant-Phos, or combinations thereof.

In another preferred example, the step carries out in the presence of a base.

In another preferred example, the alkali is selected from the group: Na₂CO₃、K₂CO₃、Cs₂CO₃、K₃PO₄, or combinations thereof.

(d) it in atent solvent, is reacted with -8 compound of Formulas I and -9 compound of Formulas I, obtains -10 compound of Formulas I；

In the above formulas, each group is as defined above.

In another preferred example, in (d) step, the atent solvent is selected from the group: methylene chloride, methanol, second Alcohol, isopropanol, n-butanol, the tert-butyl alcohol, isobutanol, dioxane, THF, DMF, DMSO, NMP, or combinations thereof.

In another preferred example, the step carries out in the presence of a base.

The application of compound of formula I

The compound of formula I can be used for one or more purposes below:

(b) the targeted inhibition agent of Bcl-2 family protein is prepared；

(c) inhibit to external non-therapeutic the activity of Bcl-2 family protein；

(d) inhibit tumor cell proliferation to external non-therapeutic；And/or

Compound of formula I of the invention can be used for preparing a kind of pharmaceutical composition, and the pharmaceutical composition includes: that (i) has The compound of formula I of effect amount or its pharmaceutically acceptable salt；(ii) pharmaceutically acceptable carrier.

In another preferred example, the effective quantity refers to therapeutically effective amount or inhibits effective quantity.

Compound of formula I of the invention can be also used for inhibiting the active method of Bcl-2 family protein, comprising steps of to suppression Object application processed inhibits a effective amount of compound of formula I as described above or its pharmaceutically acceptable salt, or applies to inhibition object With a effective amount of pharmaceutical composition as described above of inhibition.

In another preferred example, when to inhibition object application a effective amount of compound of formula I as described above of inhibition or its medicine On when acceptable salt, the inhibition effective quantity is 0.001-500nmol/L, preferably 0.01-200nmol/L.

The method that compound of formula I of the invention can be also used for extracorporeal suppression tumor cell, which comprises to inhibition Object application inhibits a effective amount of compound of formula I as described above or pharmaceutical composition as described above.

Pharmaceutical composition and method of administration

Since the compounds of this invention has the excellent inhibitory activity to Bcl-2 family protein such as Bcl-2, Bcl-xL, The compounds of this invention and its various crystal forms, pharmaceutically acceptable inorganic or organic salt, hydrate or solvate, and contain The compounds of this invention be main active pharmaceutical composition can be used for treating, prevent and alleviate by with Bcl-2 family egg White activity or the relevant disease of expression quantity.According to the prior art, the compounds of this invention can be used for treating following disease: endometrium Cancer, breast cancer, gastric cancer, bladder cancer, myeloma, liver cancer etc..

Pharmaceutical composition of the invention include safe and effective amount within the scope of the compounds of this invention or its be pharmacologically subjected to Salt and pharmacologically acceptable excipient or carrier.Wherein " safe and effective amount " refers to: the amount of compound is enough obviously Improve the state of an illness, and is unlikely to generate serious side effect.In general, pharmaceutical composition contains 1-2000mg the compounds of this invention/agent, More preferably, containing 5-200mg the compounds of this invention/agent.Preferably, described is " one " for a capsule or tablet.

" pharmaceutically acceptable carrier " refers to: one or more biocompatible solids or liquid filler or jello Matter, they are suitable for people's use and it is necessary to have enough purity and sufficiently low toxicity." compatibility " referred to herein as combines In object each component energy and the compound of the present invention and they between mutually admix, and significantly reduce the drug effect of compound.Medicine Acceptable carrier part example has cellulose and its derivates (such as sodium carboxymethylcellulose, ethyl cellulose sodium, fibre on Tie up plain acetic acid esters etc.), gelatin, talcum, solid lubricant (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oil (such as soya-bean oil, sesame Sesame oil, peanut oil, olive oil etc.), polyalcohol (such as propylene glycol, glycerol, mannitol, sorbierite), emulsifier (such as)、 Wetting agent (such as lauryl sodium sulfate), colorant, flavoring agent, stabilizer, antioxidant, preservative, apirogen water.

The method of application of the compounds of this invention or pharmaceutical composition is not particularly limited, and representative method of application includes (but being not limited to): in oral, tumor, rectum, parenteral (intravenous, intramuscular or subcutaneous) and local administration.

Solid dosage forms for oral administration includes capsule, tablet, pill, powder and granule.In these solid formulations In type, reactive compound is mixed at least one conventional inert excipients (or carrier), such as sodium citrate or Dicalcium Phosphate, or with Following compositions mixing: (a) filler or expanding material, for example, starch, lactose, sucrose, glucose, mannitol and silicic acid；(b) it bonds Agent, for example, hydroxymethyl cellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and Arabic gum；(c) moisturizer, example Such as, glycerol；(d) disintegrating agent, for example, agar, calcium carbonate, potato starch or tapioca, alginic acid, certain composition silicates, And sodium carbonate；(e) retarding solvent, such as paraffin；(f) absorbsion accelerator, for example, quaternary ammonium compound；(g) wetting agent, such as spermaceti Pure and mild glycerin monostearate；(h) adsorbent, for example, kaolin；(i) lubricant, for example, talcum, calcium stearate, tristearin Or mixtures thereof sour magnesium, solid polyethylene glycol, lauryl sodium sulfate,.In capsule, tablet and pill, dosage form also may include Buffer.

Coating and shell material preparation can be used in solid dosage forms such as tablet, sugar-pill, capsule, pill and granule, such as casing and Other materials well known in the art.They may include opacifying agent, also, reactive compound or compound in this composition Release can discharge in certain a part in the digestive tract in a delayed fashion.The example of adoptable embedding component is polymeric material And wax material.When necessary, reactive compound can also be with one of above-mentioned excipient or a variety of formation microencapsulation forms.

Liquid formulation for oral administration includes pharmaceutically acceptable lotion, solution, suspension, syrup or tincture. In addition to active compounds, liquid dosage form may include the inert diluent routinely used in this field, such as water or other solvents, increase Solvent and emulsifier, example know, ethyl alcohol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-BDO, dimethyl formyl The mixture of amine and oil, especially cottonseed oil, peanut oil, maize germ, olive oil, castor oil and sesame oil or these substances Deng.

Other than these inert diluents, composition also may include auxiliary agent, such as wetting agent, emulsifier and suspending agent, sweet taste Agent, tender taste agent and fragrance.

In addition to active compounds, suspension may include suspending agent, for example, ethoxylation isooctadecane alcohol, polyoxyethylene Sorbierite and Isosorbide Dinitrate, microcrystalline cellulose, aluminium methoxide and agar or the mixture of these substances etc..

Composition for parenteral injection may include physiologically acceptable sterile, aqueous or anhydrous solution, dispersion liquid, Suspension or lotion, and the aseptic powdery for re-dissolving into sterile Injectable solution or dispersion liquid.It is suitable aqueous and Nonaqueous carrier, diluent, solvent or excipient include water, ethyl alcohol, polyalcohol and its suitable mixture.

The dosage form of the compounds of this invention for local administration includes ointment, powder, patch, stock solution and inhalant. Active constituent aseptically with physiologically acceptable carrier and any preservative, buffer, or when necessary may need Propellant be mixed together.

The compounds of this invention can be administered alone, or be administered in combination with other pharmaceutically acceptable compounds.

It is the mammal that the compounds of this invention of safe and effective amount is applicable to treatment when using pharmaceutical composition (such as people), wherein dosage is the effective dosage pharmaceutically thought when application, for the people of 60kg weight, day is to medicament Amount is usually 1~2000mg, preferably 5~500mg.Certainly, specific dosage be also contemplated that administration route, patient health situation etc. because Element, within the scope of these are all skilled practitioners technical ability.

Main advantages of the present invention include:

1. providing a kind of compound shown in formula I.

2. providing Bcl-2 family protein inhibitor and its preparation and the application of a kind of structure novel, the inhibitor It can inhibit the activity of all kinds of Bcl-2 family proteins at much lower concentrations.

3. providing the pharmaceutical composition of a kind for the treatment of and Bcl-2 family protein activity related diseases.

Present invention will be further explained below with reference to specific examples.It should be understood that these embodiments are merely to illustrate the present invention Rather than it limits the scope of the invention.In the following examples, the experimental methods for specific conditions are not specified, usually according to conventional strip Part, or according to the normal condition proposed by manufacturer.Unless otherwise stated, otherwise percentage and number are calculated by weight.

Embodiment 1

Compound 1 (6.3g, 41.0mmol) and compound 2 (8.5g, 82.0mmol) are added to glycol monoethyl ether (40mL) In, it is stirred at reflux at 140 DEG C 18 hours.After completion of the reaction, it is concentrated to dryness, is diluted and neutralized with 0.01 mole of ammonium hydroxide, have solid Body is precipitated, and filters, and washes, dry, obtains compound 3 (5.6g, 83%).LC MS:165(M+H⁺), RT=1.10min.

Compound 3 (5.6g, 34.1mmol) is added in fuming nitric aicd (12mL) and the concentrated sulfuric acid (12mL), 100 DEG C of stirrings 1 are small Shi Hou is cooled to room temperature.Reaction solution is poured into ice water, the crude product of filtering, crude product is recrystallized with acetic acid.Compound 4 (3.48g, 49%).LC MS:210(M+H⁺), RT=1.26min.

Compound 4 (200mg, 1mmol) is added in thionyl chloride (4mL), 1 drop DMF is subsequently added into.The return stirring at 85 DEG C 3 hours.LCMS detects fully reacting, and concentration takes remaining thionyl chloride out of with toluene, obtaining compound 5, (200mg, crude product are direct For in next step).LC MS:228(M+H⁺), RT=1.58min.

Compound 6 (1.86g, 10.6mmol) and compound 7 (4.6g, 53.1mmol) are added in water (10mL), at 100 DEG C Under be stirred overnight.After completion of the reaction, it is cooled to room temperature, there is solid precipitation, filters, washing, toluene is washed, and it is dry, obtain compound 8 (2.28g, 89%).LC MS:242(M+H⁺), RT=0.30min.

By compound 8 (138mg, 0.575mmol), compound 9 (143mg, 0.575mmol) and DIEA (111mg, It 0.863mmol) is added in DMF (3mL), reaction is stirred at room temperature overnight.After completion of the reaction, it is concentrated, crude product is chromatographed through column Purifying, obtains compound 10 (142mg, 60%).LC MS:410(M+H⁺), RT=1.30min.

By compound 10 (140mg, 0.341mmol), compound 11 (65mg, 0.41mmol), palladium acetate (8mg, It 0.0341mmol) is added in DMF (2mL) and water (1mL) with sodium carbonate (55mg, 0.512mmol), it is small that reaction 20 is stirred at room temperature When.LCMS detects fully reacting, and ethyl acetate and saturated ammonium chloride is added to extract, and organic phase saturated common salt water washing is dry, mistake Filter, concentration.Crude product obtains compound 12 (133mg, 88%) through column chromatographic purifying.LC MS:442(M+H⁺), RT=1.35min.

By compound 12 (200mg, 0.45mmol), compound 5 (103mg, 0.45mmol), tetra-triphenylphosphine palladium (104mg, 0.09mmol), dry dioxane is added in cuprous iodide (34mg, 0.18mmol) and cesium carbonate (220mg, 0.68mmol) In (10mL).Microwave reaction (160 DEG C, 1 hour), LCMS detect fully reacting.Filtering, is washed with DCM:MeOH (10:1) mixed liquor It washs, crude product obtains compound 13 (92mg, 32%) through column chromatographic purifying.LC MS:633(M+H⁺), RT=1.39min.

Compound 13 (40mg, 0.06mmol), compound 14 (8mg, 0.07mmol) and triethylamine (12mg, 0.12mmol) are added Enter in ethyl alcohol (8mL).It is stirred to react at 100 DEG C 5 hours, LCMS detects fully reacting.Concentration, crude product are pure through preparation TLC Change, obtains compound 15 (15mg, 34%).LC MS:728(M+H⁺), RT=1.38min.

Compound 15:4- (4- ((4 '-chloro- biphenyl -2- base) methyl) piperazine -1- base)-N- (6- nitro -7- (((tetrahydro - 2H- pyrans -4- base) methyl) amino) quinazoline -4- base) benzsulfamide

¹H NMR(CDCl₃,400MHz)δ(ppm)9.19(s,1H),8.22(s,1H),7.95(s,1H),7.82(s,2H), 7.49(s,1H),7.35(s,6H),7.14(s,1H),6.85(m,3H),6.61(s,1H),4.01(s,2H),3.36(s,4H), 3.26(s,6H),2.48(s,4H),2.01(s,2H),1.72(s,1H),1.37(s,2H).。

Following compound can be obtained using similar approach:

4- (4- ((4 '-chloro- biphenyl -2- base) methyl) piperazine -1- base)-N- (7- (((tetrahydro -2H- pyrans -4- base) methyl) ammonia Base) -6- (trifyl) quinazoline -4- base) benzsulfamide

¹H NMR(CDCl₃,400MHz)δ(ppm)9.21(s,1H),7.96(s,1H),7.88(s,1H),7.82(s,2H), 7.48(s,1H),7.35(s,6H),7.14(s,1H),6.85(m,3H),6.59(s,1H),4.01(s,2H),3.34(s,4H), 3.23(s,6H),2.49(s,4H),2.00(s,2H),1.71(s,1H),1.36(s,2H).

4- (4- ((4 '-chloro- biphenyl -2- base) methyl) piperazine -1- base)-N- (7- ((3- morpholine base propyl) amino) -6- (trifyl) quinazoline -4- base) benzsulfamide

¹H NMR(CDCl₃,400MHz)δ(ppm)9.20(s,1H),7.96(s,1H),7.89(s,1H),7.83(s,2H), 7.48(s,1H),7.34(s,6H),7.15(s,1H),6.85(m,3H),6.58(s,1H),4.01(s,2H),3.34(s,4H), 3.23(s,6H),2.70(s,4H),2.60(m,2H),2.47(s,4H),1.68(m,2H).

4- (4- ((4 '-chloro- biphenyl -2- base) methyl) piperazine -1- base)-N- (8- nitro -7- (((tetrahydro -2H- pyrans -4- Base) methyl) amino) quinazoline -4- base) benzsulfamide

¹H NMR(CDCl₃,400MHz)δ(ppm)¹H NMR(CDCl₃,400MHz)δ(ppm)9.19(s,1H),8.02(s,1H), 7.97(s,1H),7.82(s,2H),7.35(s,6H),7.14(s,1H),7.03(s,1H),6.85(m,3H),6.58(s,1H), 4.02(s,2H),3.36(s,4H),3.26(s,6H),2.48(s,4H),2.01(s,2H),1.73(s,1H),1.37(s,2H).

4- (4- ((4 '-chloro- biphenyl -2- base) methyl) piperazine -1- base)-N- (7- (((tetrahydro -2H- pyrans -4- base) first Base) amino) -8- (trifyl) quinazoline -4- base) benzsulfamide

¹H NMR(CDCl₃,400MHz)δ(ppm)9.21(s,1H),7.98(s,1H),7.86(s,1H),7.80(s,2H), 7.35(s,6H),7.13(s,1H),7.01(s,1H),6.85(m,3H),6.60(s,1H),4.01(s,2H),3.34(s,4H), 3.23(s,6H),2.49(s,4H),2.01(s,2H),1.71(s,1H),1.35(s,2H).

4- (4- ((2- (4- chlorphenyl) -5,5- Dimethyl-cyclohex -1- alkenyl) methyl) piperazine -1- base)-N- (7- (((four Hydrogen -2H- pyrans -4- base) methyl) amino) -8- (trifyl) quinazoline -4- base) benzsulfamide

¹H NMR(CDCl₃,400MHz)δ(ppm)9.18(s,1H),7.96(s,1H),7.85(s,1H),7.80(s,2H), 7.34(s,2H),7.13(s,1H),6.99(s,1H),6.85(m,3H),6.60(s,1H),3.51(s,2H),3.33(s,4H), 3.21(s,6H),2.48(s,4H),2.06(s,2H),2.02(s,2H),1.98(s,2H),1.69(m,3H),1.35(s,2H), 1.05(s,6H).

4- (4- ((2- (4- chlorphenyl) -5,5- Dimethyl-cyclohex -1- alkenyl) methyl) piperazine -1- base)-N- (8- nitro - 7- (((tetrahydro -2H- pyrans -4- base) methyl) amino) quinazoline -4- base) benzsulfamide

¹H NMR(CDCl₃,400MHz)δ(ppm)9.20(s,1H),8.01(s,1H),7.96(s,1H),7.80(s,2H), 7.34(s,2H),7.15(s,1H),7.02(s,1H),6.86(m,3H),6.61(s,1H),3.52(s,2H),3.33(s,4H), 3.21(s,6H),2.48(s,4H),2.05(s,2H),2.01(s,2H),1.95(s,2H),1.69(m,3H),1.37(s,2H), 1.03(s,6H).

4- (4- ((2- (4- chlorphenyl) -5,5- Dimethyl-cyclohex -1- alkenyl) methyl) piperazine -1- base)-N- (7- ((3- Morpholine base propyl) amino) -8- nitro-quinazoline -4- base) benzsulfamide

¹H NMR(CDCl₃,400MHz)δ(ppm)9.19(s,1H),8.02(s,1H),7.96(s,1H),7.82(s,2H), 7.34(s,2H),7.14(s,1H),7.01(s,1H),6.86(m,3H),6.59(s,1H),3.51(s,2H),3.34(s,4H), 3.23(s,6H),2.70(s,4H),2.60(m,2H),2.47(s,4H),2.05(s,2H),2.00(s,2H),1.68(m,4H), 1.02(s,6H).

4- (4- ((2- (4- chlorphenyl) -5,5- Dimethyl-cyclohex -1- alkenyl) methyl) piperazine -1- base)-N- (7- ((2- Methoxy ethyl) amino) -8- nitro-quinazoline -4- base) benzsulfamide

¹H NMR(CDCl₃,400MHz)δ(ppm)9.20(s,1H),8.01(s,1H),7.96(s,1H),7.81(s,2H), 7.33(s,2H),7.15(s,1H),7.02(s,1H),6.86(m,3H),6.61(s,1H),3.65(m,2H),3.49(s,2H), 3.30(s,3H),3.21(s,6H),2.48(s,4H),2.05(s,2H),2.02(s,2H),1.69(m,2H),1.02(s,6H).

4- (4- ((2- (4- chlorphenyl) -5,5- Dimethyl-cyclohex -1- alkenyl) methyl) piperazine -1- base)-N- (7- ((2- Methoxy ethyl) amino) -6- nitro-quinazoline -4- base) benzsulfamide

¹H NMR(CDCl₃,400MHz)δ(ppm)9.20(s,1H),8.21(s,1H),7.96(s,1H),7.81(s,2H), 7.49(s,1H),7.33(s,2H),7.14(s,1H),6.85(m,3H),6.60(s,1H),3.65(m,2H),3.50(s,2H), 3.30(s,3H),3.22(s,6H),2.48(s,4H),2.04(s,2H),2.00(s,2H),1.69(m,2H),1.01(s,6H).

4- (4- ((2- (4- chlorphenyl) -4,4- Dimethyl-cyclohex -1- alkenyl) methyl) piperazine -1- base)-N- (7- ((2- Methoxy ethyl) amino) -6- nitro-quinazoline -4- base) benzsulfamide

¹H NMR(CDCl₃,400MHz)δ(ppm)9.19(s,1H),8.22(s,1H),7.96(s,1H),7.81(s,2H), 7.48(s,1H),7.35(s,2H),7.14(s,1H),6.83(m,3H),6.60(s,1H),3.66(m,2H),3.50(s,2H), 3.30(s,3H),3.22(s,6H),2.48(s,4H),2.05(s,2H),2.00(s,2H),1.68(m,2H),1.01(s,6H).

4- (4- ((2- (4- chlorphenyl) -4,4- Dimethyl-cyclohex -1- alkenyl) methyl) piperazine -1- base)-N- (6- nitro - 7- (((tetrahydro -2H- pyrans -4- base) methyl) amino) quinazoline -4- base) benzsulfamide

¹H NMR(CDCl₃,400MHz)δ(ppm)9.21(s,1H),8.21(s,1H),7.96(s,1H),7.81(s,2H), 7.49(s,1H),7.33(s,2H),7.14(s,1H),6.85(m,3H),6.58(s,1H),3.51(s,2H),3.32(s,4H), 3.21(s,6H),2.49(s,4H),2.03(s,2H),2.00(s,2H),1.95(s,2H),1.71(m,3H),1.37(s,2H), 1.03(s,6H).

4- (4- ((2- (4- chlorphenyl) -4,4- Dimethyl-cyclohex -1- alkenyl) methyl) piperazine -1- base)-N- (8- nitro - 7- (((tetrahydro -2H- pyrans -4- base) methyl) amino) quinazoline -4- base) benzsulfamide

¹H NMR(CDCl₃,400MHz)δ(ppm)9.19(s,1H),8.01(s,1H),7.96(s,1H),7.82(s,2H), 7.33(s,2H),7.13(s,1H),7.02(s,1H),6.84(m,3H),6.59(s,1H),3.51(s,2H),3.32(s,4H), 3.22(s,6H),2.49(s,4H),2.04(s,2H),2.00(s,2H),1.95(s,2H),1.70(m,3H),1.37(s,2H), 1.01(s,6H).

4- (4- ((2- (4- chlorphenyl) -4,4- Dimethyl-cyclohex -1- alkenyl) methyl) piperazine -1- base)-N- (7- ((2- Methoxy ethyl) amino) -8- nitro-quinazoline -4- base) benzsulfamide

¹H NMR(CDCl₃,400MHz)δ(ppm)9.18(s,1H),8.02(s,1H),7.96(s,1H),7.81(s,2H), 7.36(s,2H),7.14(s,1H),7.01(s,1H),6.84(m,3H),6.60(s,1H),3.65(m,2H),3.50(s,2H), 3.30(s,3H),3.22(s,6H),2.48(s,4H),2.05(s,2H),2.01(s,2H),1.68(m,2H),1.02(s,6H).

4- (4- ((2- (4- chlorphenyl) -4,4- Dimethyl-cyclohex -1- alkenyl) methyl) piperazine -1- base)-N- (7- ((3- Morpholine base propyl) amino) -8- nitro-quinazoline -4- base) benzsulfamide

¹H NMR(CDCl₃,400MHz)δ(ppm)9.20(s,1H),8.01(s,1H),7.96(s,1H),7.82(s,2H), 7.32(s,2H),7.14(s,1H),7.02(s,1H),6.85(m,3H),6.59(s,1H),3.51(s,2H),3.32(s,4H), 3.23(s,6H),2.69(s,4H),2.61(m,2H),2.47(s,4H),2.03(s,2H),2.00(s,2H),1.69(m,4H), 1.03(s,6H).

4- (4- ((2- (4- chlorphenyl) -4,4- Dimethyl-cyclohex -1- alkenyl) methyl) piperazine -1- base)-N- (7- (((four Hydrogen -2H- pyrans -4- base) methyl) amino) -8- (trifyl) quinazoline -4- base) benzsulfamide

¹H NMR(CDCl₃,400MHz)δ(ppm)9.18(s,1H),7.95(s,1H),7.85(s,1H),7.81(s,2H), 7.33(s,2H),7.15(s,1H),7.00(s,1H),6.85(m,3H),6.60(s,1H),3.52(s,2H),3.32(s,4H), 3.21(s,6H),2.49(s,4H),2.05(s,2H),2.00(s,2H),1.96(s,2H),1.71(m,3H),1.35(s,2H), 1.04(s,6H).

2 compound of embodiment is measured in the binding affinity of molecular level and Bcl-2 and Bcl-xL

1. test method (fluorescent polarization assay)

External Bcl-2 binding assay

The 21- residue B id BH3 peptide that 6-6- Fluoresceincarboxylic acid succinimide base ester (FAM) will be used to mark on the end N- (QEDIIRNIARHLAQVGDSMDR) (SEQ ID NO:1) is used as fluorescent marker (F1u-Bid-21).Have confirmed the fluorescent peptide With K_dFor the high binding affinity of 15.74nM.The Bcl-2 used in this measuring method is the His- fusion soluble egg of recombination It is white.

The test compound of DMSO will be dissolved in and in measurement buffer (100mM phosphoric acid clock, pH 7.5；100 μ g/ml ox bloods Clear gamma globulin, 0.02% Sodium Azide are purchased from Invitrogen Corporation, Life Technologies) in It is black that Bcl-2 protein (0.120 μM) and the 5 μ l samples of Flu-Bid-21 peptide (0.010 μM) precincubation are added to the hole Dynex 96- In color round bottom plate (Fisher Scientific), so as to generate the final volume of 125 μ 1.For each measurement, each measurement It include the binding peptide containing Bcl-2 and Flu-Bid-21 peptide (being equivalent to 0% inhibition) and containing only Flu-Bid-21 (phase on plate When in 100% inhibit) free peptide reference substance.After combining incubate within 4 hours when reaching balance, read using Ultra plate Device (Tecan U.S.Inc., Research Triangle Park, NC) is in the excitation wavelength of 485nm and the transmitted wave of 530nm Strong point measures the polarization value (mP) in terms of (mi1ipo1arization) unit that polarizes in the least.It is analyzed using nonlinear least square method IC is determined from curve graph with the curve being fitted with GraphPad Prism@software₅₀, that is, replace inhibitor when 50% binding peptide Concentration.

External BcI-xL binding assay

In order to measure the binding affinity with Bcl-xL protein, His- is recombinated using the people Bcl-xL without C- terminal hydrophobic tail The protein of label and the Bak-16mer BH3 peptide marked with 6- Fluoresceincarboxylic acid succinimide base ester (FAM).This peptide is Rendered K out_dThe binding affinity of=9.79nM.According to identical mode, use and 60nM described in Bcl-2 protein The compound of Bcl-xL and 5nM Flu-Bak peptide precincubation is containing 50mM Tris-Bis, pH 7.4；0.01% cow's serum third Being at war with property binding assay in the measurement buffer of kind globulin.

2. result

Part IC is provided in following table₅₀Data.Symbol+represent IC₅₀Less than or equal to 1uM, symbol ++ represent IC₅₀Greater than 1uM, N/A Represent temporary no data

。

Influence test of 3 compound of embodiment to acute lymphoblastic leukemia cell strain MV-4-11 ability of cell proliferation

1, test method

Compound detects the growth inhibition of acute lymphoblastic leukemia cell strain MV-4-11 cell (being purchased from ATCC cell bank) CCK-8 Cell counting Kit (Dojindo) detection.Specific step is as follows: the MV-4-11 cell in logarithmic growth phase is by conjunction Suitable density is seeded in 96 well culture plates, every 90 μ L of hole, and after overnight incubation, the compound effects 72hr of various concentration is added, and Setting solvent control group (negative control).After compound effects cell 72h, the influence of compound on intracellular proliferation uses CCK- 10 μ L CCK-8 reagents are added in 8 Cell counting Kits (Dojindo) detection, every hole, and it is small to be placed in placement 2-4 in 37 DEG C of incubators Shi Hou, with all-wave length decline orifice plate microplate reader SpectraMax 190 read, measurement wavelength be 450nm.It uses with following equation Calculate the inhibiting rate (%) of compound on tumor cell growth: inhibiting rate (%)=(OD negative control hole-OD dosing holes)/OD yin Property control wells × 100%.IC₅₀Value uses the random bundled software of microplate reader to acquire with the recurrence of four parametric methods.

2. result

The IC of part of compounds is provided in following table₅₀Value.Symbol+represent IC₅₀Less than or equal to 1uM, and symbol ++ represent IC₅₀For 1uM to 10uM, and symbol +++ represent IC₅₀Greater than 10uM

。

All references mentioned in the present invention is incorporated herein by reference, independent just as each document It is incorporated as with reference to such.In addition, it should also be understood that, after reading the above teachings of the present invention, those skilled in the art can To make various changes or modifications to the present invention, such equivalent forms equally fall within model defined by the application the appended claims It encloses.

Claims

1. a kind of such as following formula I compound represented or its pharmaceutically acceptable salt:

Wherein,

Ring A is group selected from the group below:

2. compound of formula I as described in claim 1, which is characterized in that the compound that the compound of formula I is as follows:

3. the preparation method of compound of formula I as described in claim 1, which is characterized in that comprising steps of

In the above formulas, the definition of each group is as described in the appended claim 1.

4. method as claimed in claim 3, which is characterized in that the method also includes steps:

5. the purposes of compound as described in claim 1, which is characterized in that be used for:

(b) the targeted inhibition agent of Bcl-2 family protein is prepared；

(c) inhibit to external non-therapeutic the activity of Bcl-2 family protein；

(d) inhibit tumor cell proliferation to external non-therapeutic；And/or

6. purposes as claimed in claim 5, which is characterized in that the Bcl-2 family protein is selected from the group: Bcl-2, Bcl- XL, Bcl-w, Mcl-1, Bcl2-A1, or combinations thereof；And/or

The tumour cell is leukemia cell line；Preferably lymphoblastic leukemia cell lines；It is more preferably acute leaching Bar cell leukemia cell line MV-4-11 cell.

7. a kind of pharmaceutical composition, which is characterized in that the pharmaceutical composition includes: (i) a effective amount of compound of formula I, or Its pharmaceutically acceptable salt；(ii) pharmaceutically acceptable carrier.

8. a kind of method for inhibiting Bcl-2 protein active, which is characterized in that comprising steps of to inhibiting object application to inhibit effective The compound of formula I as described in claim 1 or its pharmaceutically acceptable salt of amount, or inhibit effective quantity to inhibiting object to apply Pharmaceutical composition as claimed in claim 7.

9. a kind of method of extracorporeal suppression tumor cell, which is characterized in that the described method includes: to inhibiting object application to inhibit to have The compound of formula I as described in claim 1 or pharmaceutical composition as claimed in claim 7 of effect amount.