CN107141244B - Indolebutyric acid class inhibitors of histone deacetylase and its preparation method and application - Google Patents
Indolebutyric acid class inhibitors of histone deacetylase and its preparation method and application Download PDFInfo
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- CN107141244B CN107141244B CN201710316533.4A CN201710316533A CN107141244B CN 107141244 B CN107141244 B CN 107141244B CN 201710316533 A CN201710316533 A CN 201710316533A CN 107141244 B CN107141244 B CN 107141244B
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D209/00—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom
- C07D209/02—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom condensed with one carbocyclic ring
- C07D209/04—Indoles; Hydrogenated indoles
- C07D209/10—Indoles; Hydrogenated indoles with substituted hydrocarbon radicals attached to carbon atoms of the hetero ring
- C07D209/18—Radicals substituted by carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
- C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
- C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
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- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
- C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
- C07D417/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
Abstract
The invention belongs to field of pharmaceutical chemistry technology more particularly to indolebutyric acid histone deacetylases inhibitor and its preparation method and application.The present invention provides a kind of potent histon deacetylase (HDAC) inhibitors to further relate to its pharmaceutically acceptable salt, solvate and prodrug the present invention relates to the compound with structure formula (I).The invention further relates to pharmaceutical compositions and its pharmaceutical applications containing structure formula (I) compound.The disease of histone deacetylase activity unconventionality expression can be effectively treated,
Description
Technical field
The invention belongs to field of pharmaceutical chemistry technology more particularly to indolebutyric acid histone deacetylases inhibitor and
Preparation method and application.
Background technique
Histon deacetylase (HDAC) (HDACs) is the hydrolase of a kind of function complexity.In nucleus, wound by DNA chain
Octameric histone constitute nucleosome be constitute chromosome structural unit, histon deacetylase (HDAC) (HDACs) energy
(such as reaction equation I) is fallen into acetyl group hydrolysis on lysine residue terminal amino group in histone, just so as to cause histone
Charge density increases, and the affinity of histone and electronegative DNA is then caused to enhance, and genetic transcription is suppressed, (referring to
Christian,A.H.,et al.Curr.Opin.Chem.Biol.,1997,1,300；Kouzarides,T.,
Curr.Opin.Genet. Dev.,1999,9,40)；Wolffe,A.P.Sci.Washington,1996,272,371.In addition,
The deacetylation of nucleosome histone is also assembled with chromatin, and DNA reparation and recombination are closely related, (referring to Polo, S.E., et
al.Cancer Lett., 2005,220,1；Vidanes,G.M.,et al.Cell,2005,121,973).Recently, increasingly
More nonhistones substrates for being proved to be HDACs, such as transcription factor, cytoskeletal protein, molecular chaperones etc., (referring to
Glozak,M.A.,et al.Gene, 2005,363,15).Have the function of such complexity, its expression just because of HDACs
It is closely related with many diseases with activity imbalance, comprising: cancer, neurodegenerative disease, virus infection, inflammation, leukaemia, malaria
With diabetes etc., wherein cancer, which is undoubtedly, threatens disease the most serious to human life and health.Studies have shown that HDACs and swollen
Oncocyte occurrence and development are closely related, such as: inhibiting tumor cell differentiation and apoptosis, promote tumor cell proliferation, migration and blood vessel
Generate, enhancing tumour cell to the resistance etc. of chemotherapeutics, (referring to Witt, O., et al.Cancer Letter., 2009,
277,8)。
Reaction equation I
Have found that HDACs family there are 18 members in human body at present, according to its structure, the difference of function and distribution can divide
For four classes.Wherein, I class (HDAC1,2,3 and 8), II class (IIa:HDAC4,5,7 and 9；IIb:HDAC6,10), IV class
(HDAC11) belong to zinc ion dependence hydrolase, and Group III HDACs (SIRT 1-7) is NAD+Dependence.Studies have shown that
Closely related mainly zinc ion dependence HDACs, hdac inhibitor (HDACs Inhibitors, HDACi) energy with tumour
Effectively inhibit cancer cell multiplication, promotes Apoptosis.Moreover, HDACi has antitumor spectra wide, the low advantage of toxic side effect, they
To solid tumor, leukaemia, lymthoma all has good inhibitory activity.Therefore, for HDACs be shot design inhibitor
Hot spot as anti-tumor drug research.
The HDACi pharmacophore reported at present mostly includes following three part: Zinc Ions Chelated group (ZBG), hydrophobicity
Long-chain (Linker) and protein surface cog region (Surface Recognition Domain).Zinc Ions Chelated group can be with
The zinc ion for chelating the activated centre HDACs, to inhibit the activity of enzyme.The activity being currently known is most strong, most widely used zinc from
Sub- chelation group is hydroxamic acid group.However, many compounds in clinical research do not have good drug effect now, although
They show very excellent activity in preclinical research.Moreover, the hdac inhibitor (SAHA and FK228) of listing
Drug effect is very poor in terms for the treatment of solid tumor, and there is also half-life shorts for they, and hardly possible absorbs and pharmacokinetic profile is poor
The shortcomings that.
Summary of the invention
In view of the deficiencies of the prior art, the present invention provides a kind of inhibitors of histone deacetylase and preparation method thereof and answer
With the present invention uses hydroxamic acid group for Zinc Ions Chelated group, and indolebutyric acid is a kind of auxin, rendered
Going out antitumor potentiality, its activity can be significantly improved by being introduced into HDACi structure, and improve its lipid,
Promote the absorption of drug.
Technical scheme is as follows:
Histon deacetylase (HDAC) inhibitor with structural formula I, pharmaceutically acceptable salt, solvate or preceding
Medicine,
In general structure I:
X is-NH- ,-NHCH2-；Y is-CO- ,-CH=CHCO-；
Ar is phenyl ring, pyridine ring, thiazole ring；R is-F ,-Cl ,-OH.
Indolebutyric acid class inhibitors of histone deacetylase of the present invention, is one of following compounds:
4- (4- (1H- indol-3-yl) amide-based small)-N- hydroxybenzamide (I1)；
4- ((4- (1H- indol-3-yl) amide-based small) methylene)-N- hydroxybenzamide (I2)；
(E)-N- (4- (3- (hydroxyl amido) -3- carbonyl propyl- 1- alkene -1- base) phenyl) -4- (1H- indol-3-yl) butyryl
Amine (I3)；
4- (4- (1H- indol-3-yl) amide-based small) fluoro- N- hydroxybenzamide (I4) of -3-；
6- (4- (1H- indol-3-yl) amide-based small)-N- hydroxy nicotinoyl amine (I5)；
4- (4- (1H- indol-3-yl) amide-based small) chloro- N- hydroxybenzamide (I6) of -3-；
3- (4- (1H- indol-3-yl) amide-based small)-N- hydroxybenzamide (I7)；
4- (4- (1H- indol-3-yl) amide-based small)-N, 3- dihydroxy benzoyl amine (I8)；
2- (4- (1H- indol-3-yl) amide-based small)-N- hydroxyl thiazole -4-carboxamide (I9).
The present invention also provides these compounds to prevent or treat and histone deacetylase activity unconventionality expression phase
Application in the drug of the mammalian diseases of pass.The lactation related to histone deacetylase activity unconventionality expression
Animal diseases include: cancer, neurodegenerative disease, virus infection, inflammation and diabetes etc..
Therefore, the invention further relates to the pharmaceutical compositions containing Compounds of structural formula I.
Detailed description of the invention
Definition and term used
Term and definition meaning used herein is as follows:
" pharmaceutically acceptable salt " refers to that compound has curative effect and nontoxic salt form.It can be by any acidic-group
(such as carboxyl) forms anion salt, or forms cationic salts by any basic group (such as amino).It is known in the art it is many in this way
Salt.The cationic salts formed on any acidic-group (such as carboxyl), or formed on any basic group (such as amino)
Anion salt.These salt there are many be it is known in the art, if cationic salts include that alkali metal (such as sodium and potassium) and alkaline earth are golden
Belong to the salt and organic salt (such as ammonium salt) of (such as magnesium and calcium).It can also be convenient by using (I) of corresponding acid processing alkaline form
Ground obtains anion salt, and such acid includes inorganic acid such as sulfuric acid, nitric acid, phosphoric acid etc.；Or organic acid such as acetic acid, propionic acid, hydroxyl
Acetic acid, 2 hydroxy propanoic acid, Acetylformic acid, oxalic acid, malonic acid, succinic acid, maleic acid, fumaric acid, malic acid, tartaric acid, 2-
Hydroxyl -1,2,3- the third three acid, methanesulfonic acid, ethanesulfonic acid, benzene methanesulfonic acid, 4- toluenesulfonic acid, cyclohexyl sulfinic acid, 2- hydroxy benzenes first
Acid, 4- amino-2-hydroxybenzoic acid etc..These salt are known to those of skill in the art, and those skilled in the art can prepare ability
Any salt provided by domain knowledge.In addition, those of skill in the art can take certain according to factors such as solubility, stability, easy preparations
It plants salt and gives up another salt.The measurement of these salt and optimization are in the experience range of those of skill in the art.
" prodrug " refer to that drug obtains after modifying for chemical structure it is inactive in vitro or active it is smaller, pass through in vivo
The conversion of enzyme or non-enzymatic releases active medicine and plays the compound of drug effect.
Structure formula (I) compound can by it is other it is protected in the form of or derivative in the form of exist, these forms to this
It is it will be apparent that should be included within the scope of the present invention for the technical staff of field.
The preparation method of the indolebutyric acid class inhibitors of histone deacetylase, reaction step and reaction equation are as follows:
Preparation method includes the following steps:
Synthetic route:
Using 3- indolebutyric acid as raw material, advanced condensation reaction is re-introduced into hydroxamic acid and obtains final product, and reaction equation is such as
Under:
Reagent in 1 reaction equation of said synthesis route: (1) TBTU, corresponding amino acid methyl ester, Et3N, DCM；(2) NHOK,
CH3OH。
The intermediate of the inhibitors of histone deacetylase is prepared, intermediate includes: methyl 4- (4- (1H- indoles -3-
Base) amide-based small) benzoic ether, methyl 4- ((4- (1H- indol-3-yl) amide-based small) methylene) benzoic ether, (E)-first
Base 3- (4- (4- (1H- indol-3-yl) amide-based small) phenyl) acrylate, methyl 4- (4- (1H- indol-3-yl) butyramide
Base) -3- fluorobenzoate, methyl 6- (4- (1H- indol-3-yl) amide-based small) nicotinate, methyl 4- (4- (1H- indoles -3-
Base) amide-based small) -3- chlorobenzoic acid ester, methyl 3- (4- (1H- indol-3-yl) amide-based small) benzoic ether, methyl 4- (4-
(1H- indoles -3- base) amide-based small) -3- hydroxybenzoate, methyl 2- (4- (1H- indol-3-yl) amide-based small) thiazole -
4- formic acid esters.
Those skilled in the art can change above-mentioned steps to improve yield, they can knowing substantially according to this field
Know the route for determining synthesis, such as selects reactant, solvent and temperature, it can be by using various GPF (General Protection False bases to avoid secondary anti-
The generation answered is to improve yield.These conventional guard methods can be found in such as T.Greene, Protecting Groups
in Organic Synthesis.。
Due to the high homology of each hypotype catalytic center of zinc ion dependence histon deacetylase (HDAC) (HDACs), I
Select the Hela cell extract (comprising HDAC1, HDAC2, HDAC3 and HDAC8) containing histon deacetylase (HDAC) come into
The test of row enzymatic activity.HDACs active fluoro analysis method (two-step method), energy is quick, facilitates detection HDACs activity, and operation is simple
It is single, high sensitivity.The first step, the lysine HDACs fluorogenic substrate (Boc-Lys (acetyl)-containing an acylated chains
AMC), with the Hela cell extract sample incubation containing histon deacetylase (HDAC), make substrate deacetylate, activate bottom
Object.Second step generates this fluorophor (i.e. chromophore) of AMC, in launch wavelength/excitation with pancreatin hydrolysis Boc-Lys-AMC
Wavelength (390nm/460nm) measures fluorescence intensity, so that inhibiting rate is calculated according to inhibitor group and the fluorescence intensity of control group, and
It asks and calculates IC50 value.Enzymatic activity test philosophy is shown in reaction equation II.
The test of the cell activity of compound uses Thiazolyl blue detection method (mtt assay), human tissue cell's lymphoma cell
Strain (U937), human erythroleukemia cell's strain (K562), the cell suspension of people's acute leukemia cells strain (HL60) are inoculated in respectively
96 orifice plates, the culture medium that the compound containing various concentration is added in every hole is dyed after being incubated for MTT, after continuing incubation, in enzyme
The absorbance (OD value) for measuring every hole on instrument at 570nm is marked, inhibitory rate of cell growth is calculated, so that it is determined that the work of compound
Property.
The external suppression enzyme test of the compound of logical formula (I) proves such compound for the suppression of effective histon deacetylase (HDAC)
Preparation.
Pharmaceutical composition containing the compounds of this invention
Partial derivatives of the invention can exist in a free form or in the form of salts.Manyization known to those skilled in the art
Pharmaceutically acceptable salt of polymer type and preparation method thereof.Pharmaceutically acceptable salt includes conventional avirulent salt,
The quaternary ammonium salt formed including such chemical combination alkaloids and inorganic or organic acid.
The compound of the present invention can form hydrate or solvate.It is known to those skilled in the art by compound and water one
It rises and is formed by hydrate or the in the solution method with formation solvate when the concentration of suitable organic solvent when freeze-drying.
The present invention includes the drug containing therapeutic dose the compounds of this invention and one or more pharmaceutically acceptable carriers
And/or the pharmaceutical composition of excipient.Carrier includes such as salt water, buffered saline, glucose, water, glycerol, the knot of ethyl alcohol and they
Object is closed, is more fully hereinafter discussed.If desired, the composition can also include small amount of wetting agent or emulsifier or pH
Buffer.The composition can be liquid, suspension, emulsion, tablet, pill, capsule, extended release preparation or powder.The group
Suppository can be configured to traditional binder and carrier such as triglyceride by closing object.Oral preparation may include standard vector such as
The mannitol of drug grade, lactose, starch, magnesium stearate, saccharin sodium, cellulose and magnesium carbonate etc..Optionally preparation and
Fixed, preparation can design mixing, granulation and compression or solvent components.In another approach, the composition can be configured to receive
Rice grain.
The pharmaceutical carrier used can be solid or liquid.
Typical solid carrier includes lactose, land plaster, sucrose, talcum, gel, agar, pectin, Arabic gum, tristearin
Sour magnesium, stearic acid etc..Solid carrier may include it is one or more may be used as fumet simultaneously, lubricant, solubilizer hangs
Floating agent, filler, glidant, compression aid, the substance of adhesive or tablet-disintegrating agents；It may also is that encapsulating material.In powder
In, carrier is the solid of fine crushing, the mixing of it and the active constituent of fine crushing.In tablets active constituent with have must
The carrier for the compression property wanted mixes in proper proportions, is compressed with the shapes and sizes of needs.Powder and tablet preferably comprise
At most 99% active constituent.Suitable solid carrier includes, for example, calcium phosphate, magnesium stearate, talcum, sugar, lactose, dextrin form sediment
Powder, gel, cellulose, methylcellulose, sanlose, polyvinylpyrrolidone alkanone, low melt wax and ion
Exchanger resin.
Typical liquid-carrier includes syrup, peanut oil, olive oil, water etc..Liquid-carrier is used to prepare solution, suspends
The composition of liquid, emulsion, syrup, tincture and sealing.Active constituent can dissolve or be suspended in pharmaceutically acceptable liquid and carry
Body such as water, organic solvent, the mixture of the two or pharmaceutically acceptable oils or fat.Liquid-carrier may include other conjunctions
Suitable medicated premix such as solubilizer, emulsifier, buffer, preservative, sweetener, fumet, suspending agent, thickener, pigment,
Viscosity modifier stablizes shape or osmo-regulators.Suitable example packet for oral and parenteral administration liquid-carrier
Include water (partly comprising such as above-mentioned additive, such as cellulose derivative, preferably carboxymethyl cellulose sodium salt solution), alcohol
(including monohydric alcohol and polyalcohol, such as ethylene glycol) and their derivative and oils (such as fractionated coconut oil and peanut oil).
Carrier for parenteral administration can also be grease such as ethyl oleate and isopropyl myristate.Sterile liquid-carrier is used
In the sterile fluid composition of parenteral administration.Liquid-carrier for pressurized compositions can be halogenated hydrocarbons or other pharmacy
Upper acceptable propellant.Sterile solution or aaerosol solution composition of liquid medicine can be used to, for example, intravenously, intramuscular, abdomen
In film or it is subcutaneously injected.It single push-in or can be gradually injected when injection, enter perfusion in 30 minutes passages through which vital energy circulates.The compound can be with
It is administered orally in the form of liquid or solid composite.
Carrier or excipient may include time delay material known in the art, such as glycerin monostearate or distearyl
Acid glyceride may also include wax, ethyl cellulose, hydroxypropyl methyl cellulose, methylmethacrylate etc..When preparation is used for mouth
When taking, it is recognized that (phosphatide (phospholipid) and 1,2-PD are concentrated PHOSALPG-50, A.Nattermann&Cie.
GmbH 0.01% Tween 80 in) is used for the preparation of the acceptable oral preparation of other compounds, is adapted to the present invention
The preparation of various compounds.
Miscellaneous medicament forms can be used when giving the compounds of this invention.If preparation can using solid carrier
Think tablet, the powder or piller form or pastille or Lozenge forms being placed into hard capsule.The amount of solid carrier is very big
Change in degree, it is preferred that from about 25mg to about 1.0g.If preparation can be syrup, emulsion, flexible glue using liquid-carrier
Capsule, aseptic injectable solution or suspension in ampoule or bottle or non-aqueous liquid suspension.
In order to obtain stable water-soluble dosage form, compound or its pharmaceutically acceptable salt can be dissolved in it is organic or
The aqueous solution of inorganic acid, 0.3M succinic acid or citric acid solution.Selectively, acid derivative can be dissolved in suitable alkalinity
Solution.If cannot get soluble form, compound can be dissolved in suitable cosolvent or their combination.It is such suitable total
The example of solvent includes but are not limited to, ethyl alcohol of the concentration range from 0-60% total volume, propylene glycol, Liquid Macrogol,
Polysorbate 80, glycerol, polyoxyethylene fatty acid ester, fatty alcohol or glycerol hydroxy fatty acid ester etc..
Various release systems are administration that is known and can be used for compound or other various preparations, these preparation packets
Include tablet, capsule, the solution of injectable, the capsule in liposome, particle, microcapsules, etc..The method of introducing include but not
It is confined to skin, it is intradermal, it is intramuscular, it is intravenous in peritonaeum, it is subcutaneous, in nasal cavity, lung, it is peridural, eyes
(generally preferable) oral route.Compound can be administered by any convenient or other approach appropriate, such as logical
Cross injection or bolus injection, by epithelium or mucous membrane route (for example, oral mucosa, rectum and intestinal mucosa, etc.) absorb or
It can be administered by the bracket of carrying medicament and together in other biological activities agent.It can be administered either systemically or locally.For nose,
When the treatment or prevention of bronchus or lung disease, preferred administration route is oral, nasal administration or bronchus smoke agent or spraying
Device.
Indolebutyric acid class compound in the present invention is better than positive control drug to the inhibitory activity of histon deacetylase (HDAC),
With good development prospect, and it can be used as the lead compound for finding new and effective histon deacetylase (HDAC) inhibitor.This
Outside, such compound exhibits improvements over positive control Vorinostat's (SAHA) in the test of anti-tumour cell proliferative in vitro
Activity has good development prospect.
Detailed description of the invention
Attached drawing 1 is that HDACs active fluoro analysis method tests enzymatic activity, and wherein Histone deacetylase is group egg
White deacetylase, Trypsin are trypsase, and 4-amino-7-methylcoumarin is 4- amino -7- methylcoumarin
Element.
Specific embodiment
The present invention will be further explained with reference to the examples below, but not limited to this.
The synthesis of 1. the compounds of this invention of embodiment
Methyl 4- (4- (1H- indol-3-yl) amide-based small) benzoic ether
Indolebutyric acid (1.0g, 5mmol) is dissolved in 25mL tetrahydrofuran, is added triethylamine (0.55g, 5.5mmol), is added
O- benzotriazole-N, N, N', N'- tetramethylurea tetrafluoro boric acid (TBTU) (1.8g, 5.5mmol).Room temperature reaction 20 minutes
Afterwards, methyl p-aminobenzoate hydrochloride (0.94g, 5mmol) is added, then plus triethylamine (0.5g, 5mmol).It is small to react at room temperature 6
The tetrahydrofuran in reaction solution is evaporated off in Shi Hou, with ethyl acetate lysate, uses the citric acid solution of 1mol/L, saturation respectively
Sodium bicarbonate solution, saturated salt solution respectively wash 3 times, and anhydrous magnesium sulfate is dry, and solvent evaporated obtains crude product, and crude product is through ethyl acetate
Recrystallize to obtain the shallow white solid of 1.1g.Yield: 42%, ESI-MS m/z:337.4 [M+H+]。
4- (4- (1H- indol-3-yl) amide-based small)-N- hydroxybenzamide (I1)
Azanol potassium (NH2OK) the preparation of solution: the saturation absolute methanol solution of 14mL potassium hydroxide is added drop-wise to 24mL and contains
In the absolute methanol solution of 4.67g (67mmol) hydroxylamine hydrochloride, controls interior temperature and be lower than 40 DEG C, be added dropwise, cooling reaction solution, filter
Except white potassium chloride precipitates, gained filtrate is closed to be saved backup.
Methyl 4- (4- (1H- indol-3-yl) amide-based small) benzoic ether (0.67g, 2mmol) is dissolved in 10mL anhydrous methanol
Afterwards, the above-mentioned azanol potassium (NH of 3.5mL is added thereto2OK) solution.After 0.5 hour, methanol, the hydrochloric acid solution acid of 2mol/L is evaporated off
Change to pH3-4, be then extracted with ethyl acetate, saturated common salt water washing is used after combined ethyl acetate layer, it is dry through anhydrous magnesium sulfate
Dry, solvent evaporated obtains crude product, and crude product obtains 0.34g white powder through re-crystallizing in ethyl acetate.Yield: 49%.ESI-MS m/z:
338.4[M+H+]。1H NMR(400MHz,DMSO)δ11.07(s,1H),10.77(s,1H),10.08 (s,1H),8.91(s,
1H), 7.70-7.63 (m, 4H), 7.52 (d, J=7.9Hz, 1H), 7.33 (d, J=8.1Hz, 1H), 7.13 (d, J=2.1Hz,
1H), 7.05 (dd, J=11.1,4.0Hz, 1H), 6.96 (dd, J=10.9,3.9Hz, 1H), 2.72 (dd, J=20.2,
12.8Hz, 2H), 2.38 (dd, J=20.0,12.6Hz, 2H), 2.12-1.81 (m, 2H)
The synthesis of 2. the compounds of this invention of embodiment
Methyl 4- ((4- (1H- indol-3-yl) amide-based small) methylene) benzoic ether
Indolebutyric acid (1.0g, 5mmol) is dissolved in 25mL tetrahydrofuran, is added triethylamine (0.55g, 5.5mmol), is added
O- benzotriazole-N, N, N', N'- tetramethylurea tetrafluoro boric acid (TBTU) (1.8g, 5.5mmol).Room temperature reaction 20 minutes
Afterwards, methyl 4- (amino methyl) benzoic ether hydrochloride (1.0g, 5mmol) is added, then plus triethylamine (0.5g, 5mmol).Room temperature
After reaction 6 hours, the tetrahydrofuran in reaction solution is evaporated off, it is molten with the citric acid of 1mol/L respectively with ethyl acetate lysate
Liquid, saturated sodium bicarbonate solution, saturated salt solution respectively wash 3 times, and anhydrous magnesium sulfate is dry, and solvent evaporated obtains crude product, crude product warp
Re-crystallizing in ethyl acetate obtains the shallow white solid of 1.3g.Yield: 57%, ESI-MS m/z:351.4 [M+H+]。
4- ((4- (1H- indol-3-yl) amide-based small) methylene)-N- hydroxybenzamide (I2)
Methyl 4- ((4- (1H- indol-3-yl) amide-based small) methylene) benzoic ether (0.81g, 2mmol) is dissolved in 10mL
After anhydrous methanol, the above-mentioned azanol potassium (NH of 3.5mL is added thereto2OK) solution.After 0.5 hour, methanol is evaporated off, 2mol/L's
Hydrochloric acid solution is acidified to pH3-4, is then extracted with ethyl acetate, and saturated common salt water washing is used after combined ethyl acetate layer, through nothing
Water magnesium sulfate is dry, and solvent evaporated obtains crude product, and crude product obtains 0.41g white powder through re-crystallizing in ethyl acetate.Yield: 48%.
ESI-MS m/z:352.4 [M+H+]。1H NMR(500MHz,dmso)δ11.15(s,2H),10.74(s,1H), 8.98(s,
2H), 8.36 (t, J=5.9Hz, 1H), 7.79 (dd, J=96.4,8.2Hz, 2H), 7.45 (t, J=20.7Hz, 1H), 7.32
(dd, J=21.5,12.1Hz, 3H), 7.09 (d, J=1.5Hz, 1H), 7.04 (t, J=7.5Hz, 1H), 6.94 (dd, J=
18.3,11.1Hz, 1H), 4.44-4.16 (m, 2H), 2.67 (t, J=7.5Hz, 2H), 2.24 (dd, J=34.7,27.3Hz,
2H),1.96–1.80(m,2H).
The synthesis of 3. the compounds of this invention of embodiment
(E)-methyl 3- (4- (4- (1H- indol-3-yl) amide-based small) phenyl) acrylate
Indolebutyric acid (1.0g, 5mmol) is dissolved in 25mL tetrahydrofuran, is added triethylamine (0.55g, 5.5mmol), is added
O- benzotriazole-N, N, N', N'- tetramethylurea tetrafluoro boric acid (TBTU) (1.8g, 5.5mmol).Room temperature reaction 20 minutes
Afterwards, (E)-methyl 3- (4- aminophenyl) acrylate (1.1g, 5mmol), then plus triethylamine (0.5g, 5mmol) is added.Room temperature
After reaction 6 hours, the tetrahydrofuran in reaction solution is evaporated off, it is molten with the citric acid of 1mol/L respectively with ethyl acetate lysate
Liquid, saturated sodium bicarbonate solution, saturated salt solution respectively wash 3 times, and anhydrous magnesium sulfate is dry, and solvent evaporated obtains crude product, crude product warp
Re-crystallizing in ethyl acetate obtains the shallow white solid of 1.2g.Yield: 49%, ESI-MS m/z:363.4 [M+H+]。
(E)-N- (4- (3- (hydroxyl amido) -3- carbonyl propyl- 1- alkene -1- base) phenyl) -4- (1H- indol-3-yl) butyryl
Amine (I3)
(E)-methyl 3- (4- (4- (1H- indol-3-yl) amide-based small) phenyl) acrylate (0.82g, 2mmol) is dissolved in
After 10mL anhydrous methanol, the above-mentioned azanol potassium (NH of 3.5mL is added thereto2OK) solution.After 0.5 hour, methanol, 2mol/ is evaporated off
The hydrochloric acid solution of L is acidified to pH3-4, is then extracted with ethyl acetate, and saturated common salt water washing is used after combined ethyl acetate layer,
Dry through anhydrous magnesium sulfate, solvent evaporated obtains crude product, and crude product obtains 0.35g white powder through re-crystallizing in ethyl acetate.Yield:
44%.ESI-MS m/z:364.4 [M+H+]。1H NMR(500MHz,dmso)δ10.76(s,1H),10.68(s,1H), 10.03
(s, 1H), 8.98 (s, 1H), 7.64 (d, J=8.3Hz, 2H), 7.50 (dd, J=16.6,8.1Hz, 3H), 7.38 (d, J=
15.7Hz, 1H), 7.32 (d, J=8.1Hz, 1H), 7.11 (d, J=15.2Hz, 1H), 7.06 (dd, J=16.8,9.5Hz,
1H), 6.96 (t, J=7.4Hz, 1H), 6.34 (d, J=15.8Hz, 1H), 2.88-2.61 (m, 2H), 2.38 (t, J=
7.4Hz,2H), 2.03–1.81(m,2H).
4 target compound inhibition of histone deacetylase activity test (In vitro) of embodiment
Histon deacetylase (HDAC) (HDACs) active fluoro analysis method is mainly in two steps: the first step contains an acetylation
The lysine HDACs fluorogenic substrate (Boc-Lys (acetyl)-AMC) of side chain is thin with the Hela containing histon deacetylase (HDAC)
Born of the same parents' extract sample (including HDAC1, HDAC2, HDAC3 and HDAC8) is incubated for, and is made substrate deacetylate, is activated substrate.Second
Step generates this fluorophor (i.e. chromophore) of AMC, in launch wavelength/excitation wavelength with pancreatin hydrolysis Boc-Lys-AMC
(390nm/460nm) measures fluorescence intensity, to calculate inhibiting rate according to inhibitor group and the fluorescence intensity of control group, and asks calculation
IC50Value.Enzymatic activity test philosophy is shown in patent specification part related content.Experimental result is shown in Table 1.
Table 1. presses down enzyme test result in vitro
| Compound | I1 | I2 | I3 | I4 | I5 | I6 | I7 | I8 | I9 | SAHA |
| IC50(μM) | 0.14 | 0.21 | 0.19 | 0.22 | 0.35 | 0.17 | 0.19 | 0.25 | 0.46 | 1.05 |
SAHA trade name Zolinza, general entitled Vorinostat are U.S. Food and Drug Administration (FDA)
In the histon deacetylase (HDAC) inhibitor of approval listing in 2006.
Above-mentioned test result shows that indolebutyric acid class compound shows inhibition stronger to histon deacetylase (HDAC) and lives
Property, and positive control drug Vorinostat (SAHA) is better than to the inhibitory activity of histon deacetylase (HDAC) in testing, tool
There is good development prospect, and can be used as the lead compound for finding new and effective histon deacetylase (HDAC) inhibitor.
The activity test (In vitro) of 5 target compound of embodiment inhibition cell Proliferation
The external activity test for inhibiting cancer cell multiplication, knot are carried out to indolebutyric acid histone deacetylases inhibitor
Fruit is shown in Table 2.
Term explanation:
U937: human tissue cell's lymphoma cell strain.
K562: human erythroleukemia cell's strain.
HL60: people's acute leukemia cells strain.
SAHA: trade name Zolinza, general entitled Vorinostat are U.S. Food and Drug Administration (FDA)
In the histon deacetylase (HDAC) inhibitor of approval listing in 2006.
DMSO: dimethyl sulfoxide.
IC50: half-inhibitory concentration.
[1. material] U937, K562, HL60 cell strain, Methyl thiazoly tetrazolium assay MTT, 10% fetal calf serum, 96 orifice plates.
[2. method]
Tri- kinds of tumor cell lines of cell culture U937, K562, HL60 all use routine culture.Test Shi Junyong logarithmic growth
Phase cell.
Cell growth detection (mtt assay) U937, K562, HL60 cell suspension is adjusted to 1 × 105/ ml, is inoculated in respectively
96 orifice plates (50 hole μ l/), 5000 cells/wells.After bed board 4h, the culture of 50ul compound containing various concentration is added in every hole
Base makes final compound concentration in hole be respectively as follows: 1000,200,40,8,1.6,0.32ug/ml, and each concentration sets three multiple holes, no
Make blank when the hole reading of refinement born of the same parents, compound blank well is made in the hole that compound is not added in refinement born of the same parents, and it is right that SAHA makees the compound positive
According to.48h is incubated in 37 DEG C, 5% carbon dioxide, the MTT dyeing liquor of 10 μ l 0.5% is added in every hole, continue after being incubated for 4h,
2500rpm is centrifuged 30min, then abandons culture medium in plate hole, and dimethyl sulfoxide, the hole 200ul/ is added.In microplate reader in
The absorbance OD value in every hole is measured at 570nm, inhibitory rate of cell growth is calculated as follows:
2 cell proliferation experiment result of table
aNumerical value is the average value tested three times in table, and the numerical tabular after " ± " shows standard deviation.
Upper table test data shows multiple indolebutyric acid histone deacetylases inhibitors antitumor cell in vitro
The activity of positive control SAHA is exhibited improvements in the test of proliferation, and there is good development prospect.
Claims (7)
1. indolebutyric acid class inhibitors of histone deacetylase, chemical structure with structure formula (I) and its can pharmaceutically connect
The salt received,
In general structure I:
X is-NH- ,-NHCH2-；Y is-CO-；
Ar is phenyl ring, pyridine ring, thiazole ring；R is-F ,-Cl ,-OH.
2. indolebutyric acid class inhibitors of histone deacetylase, it is characterised in that: be one of following compounds:
4- (4- (1H- indol-3-yl) amide-based small)-N- hydroxybenzamide (I1)；
4- ((4- (1H- indol-3-yl) amide-based small) methylene)-N- hydroxybenzamide (I2)；
4- (4- (1H- indol-3-yl) amide-based small) fluoro- N- hydroxybenzamide (I4) of -3-；
6- (4- (1H- indol-3-yl) amide-based small)-N- hydroxy nicotinoyl amine (I5)；
4- (4- (1H- indol-3-yl) amide-based small) chloro- N- hydroxybenzamide (I6) of -3-；
3- (4- (1H- indol-3-yl) amide-based small)-N- hydroxybenzamide (I7)；
4- (4- (1H- indol-3-yl) amide-based small)-N, 3- dihydroxy benzoyl amine (I8)；
2- (4- (1H- indol-3-yl) amide-based small)-N- hydroxyl thiazole -4-carboxamide (I9).
3. the preparation method of indolebutyric acid class inhibitors of histone deacetylase as claimed in claim 1 or 2, it is characterised in that: packet
Include following steps:
Using 3- indolebutyric acid as raw material, condensation reaction is first carried out, hydroxamic acid is re-introduced into and obtains final product, reaction equation is as follows:
Reagent in 1 reaction equation of said synthesis route: (1) TBTU, corresponding amino acid methyl ester, Et3N, DCM；(2) NHOK, CH3OH。
4. the preparation method of indolebutyric acid class inhibitors of histone deacetylase according to claim 3, it is characterised in that:
The intermediate of the indolebutyric acid class inhibitors of histone deacetylase is prepared, intermediate includes: methyl 4- (4- (1H- indoles-
3- yl) amide-based small) benzoic ether, methyl 4- ((4- (1H- indol-3-yl) amide-based small) methylene) benzoic ether, methyl
4- (4- (1H- indol-3-yl) amide-based small) -3- fluorobenzoate, methyl 6- (4- (1H- indol-3-yl) amide-based small) cigarette
Acid esters, methyl 4- (4- (1H- indol-3-yl) amide-based small) -3- chlorobenzoic acid ester, methyl 3- (4- (1H- indol-3-yl) fourth
Amide groups) benzoic ether, methyl 4- (4- (1H- indol-3-yl) amide-based small) -3- hydroxybenzoate, methyl 2- (4- (1H-
Indol-3-yl) amide-based small) 4-thiazolecarboxylic acid ester.
5. indolebutyric acid class inhibitors of histone deacetylase of any of claims 1 or 2 is in preparation prevention or treatment and group egg
Application in the drug of the relevant mammalian diseases of white deacetylation enzymatic activity unconventionality expression.
6. application according to claim 5, it is characterised in that: described with histone deacetylase activity unconventionality expression
Related mammalian disease include: cancer, neurodegenerative disease, virus infection, inflammation and diabetes.
7. it is a kind of suitable for the oral pharmaceutical composition for giving mammal, include indolebutyric acid class group described in claim 1,2
Albumen deacetylase inhibitor and one or more pharmaceutically acceptable carriers or excipient.
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| CN103467359A (en) * | 2013-09-27 | 2013-12-25 | 山东大学 | Cinnamon amides histone deacetylase inhibitor with benzpyrole and preparation method and application of same |
| CN105348169A (en) * | 2015-11-16 | 2016-02-24 | 青岛大学 | Histone deacetylase inhibitor (E)-3-(2-(1-(4-chlorobenzoyl)-5-methoxy-2-methyl-1H-indole-3-yl)acetamido)-N-hydroxy-butyl-2-enamide and its preparation method and use |
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