CA-4 cyclen derivatives and its antitumor properties
Technical field
The present invention relates to pharmaceutical technology field, and in particular to CA-4 Macrocyclic polyamine analog derivative and its metal complex,
Or such compound pharmaceutically acceptable salt, individually or optionally combine use with other one or more pharmaceutically active compounds
In the related disease for the treatment of tumour.
Background technology
Macrocyclic polyamine is the important ligand molecular of a class, with its variable steric configuration and electronic structure, and result in one is
Row special nature.Because the nitrogen-atoms of introducing has special coordination compatibility, institute's shape to transition metal ions, heavy metal ion
Into macrocyclic complex in host-guest chemistry, chemical simulation, molecular magnet, molecular recognition and information transfer have important meaning
Justice, but also can be by the use of some metal ion match with specific structure as Model Molecule, for research vital movement
Process provides extensive possibility.
In structure, Macrocyclic polyamine complex and the macrocyclic complex existed in organism, the cooperation of such as porphyrin and corrin
Thing, with similitude, compared with general crown compound, Macrocyclic polyamine has two key characters:Macrocyclic polyamine and many gold
Category ion can form stable complex, and ring member nitrogen atoms have a strong basicity, thus can and H+Form ammonium ion;It is easier to import on ring
Carboxyl, hydroxyl, phosphonate group and other hydrophilic radicals or functional group, can obtain a variety of water miscible parts.Macrocyclic polyamine
One critically important purposes of metal complex is applied in enzyme simulation research, as carboxylate, phosphate and nucleic acid
The research of catalyzing hydrolysis.Macrocyclic polyamine molecule can form stable complex with many metal ions, and ring member nitrogen atoms have strong
Alkalescence, energy and H+Ammonium salt is formed, is easier to import carboxyl, hydroxyl, phosphonate group and other hydrophilic radicals or functional group, can obtain
A series of water soluble ligands.The research of this year, Macrocyclic polyamine and its metal complex is more and more, reports in succession both at home and abroad
The synthesis of all kinds of cyclen derivatives and its metal complex and property.The larger macrocyclic polyamine compound of current application prospect
Mainly have:Isosorbide-5-Nitrae, 7- 7-triazacyclononanes (1), Isosorbide-5-Nitrae, 7,10- tetraazacyclododecanands (cyclen, 2), Isosorbide-5-Nitrae, 8,11- tetra- nitrogen
The heterocycle tetradecane (cyclam, 3).
The elementary cells such as macrocyclic tetraaza part and the vitamin B12 in organism, chlorophyll are similar, many of life process
It is oxidation-reduction process using metal ion match as reaction center that step, which can be assumed that, and wherein macrocyclic tetraaza complex is normal
As the analogies of some enzymatic structure units in organism, therefore research macrocyclic tetraaza complex is significant.Big ring is more
Study that must to compare many be complex using Cyclen as parent in amine metal complex.
Macrocyclic polyamine part has the performance that DNA is cut, with certain anti-cancer function.The Cyclen of relatively early research coordinates
Super coiled DNA (pBR322) can be cut into the type of incising and linear DNA by thing under aerobic conditions, and pharmacological evaluation shows that this is matched somebody with somebody
Compound has active anticancer to P388 and BEL-7404 cell lines in vivo, and has interference in the G0-G1 stages to K562 cancerous cell lines
Effect.Hereafter, a variety of functional groups are connected on nitrogen-atoms of the researcher on ring so that the research of Cyclen complexs
It is more deep.The complex formed after new group is all connected on Cyclen four nitrogen-atoms, in H2O2In the presence of, Neng Gouchong
When single-stranded and double-strand scissors, super coiled DNA is cut into the type of incising and linear DNA.It is common as the electron paramagnetic of trapping bait using DMPO
Result of shaking proves that cutting mechanism may be free radical mechanism.
In addition, Macrocyclic polyamine complex also has the function of Supramolecular Recognition, nucleotides and DNA, RNA can be known
Not.Macrocyclic tetraamines such as l, 4,7, lO- tetraazacyclododecanands (Cyclen) and derivative and Zn2+The complex of formation, not only exists
Very unique activity is shown in terms of hydrolyse phosphate esters, and is also extremely to have in terms of identification nucleotides and DNA, RNA
Effect.The Cyclen complexs of Zn (II) one with side arm aromatic ring can pass through Zn2+The formation of-imido grpup anion key, there is selection
Combined with T or U in single-screw and duplex DNA or RNA to property, the work(of molecule zipper lock is played the part of in the A-T chains of DNA breakage
Energy.The complex of macrocyclic tetraamines compound with one or two arylmethyl group can selectively with natural duplex DNA
(containing 150 base-pairs) connection, with reference to generation in A_T rich regions.This selectivity and binding ability are fixed against aromatic ring π-π
Stacking ability and number of aromatic rings
And combretastatin A4 (its structure is as shown in Equation 4) is two phenyl ring connected by ethylenic linkage, with powerful π-π
Accumulation ability.It also has antitumor activity simultaneously, can directly act on endothelial cell, the endothelial cell of proliferative induction withers
Die, so as to suppress tumor neovasculature generation.
Therefore, the present invention develops a kind of new antineoplastic, and Macrocyclic polyamine complex and CA-4 are passed through into chemical bond
Organically it is combined together.It is desirable that the DNA identifications of Macrocyclic polyamine complex and dissection and CA-4 in the compound
Microtubulin-resisting synthesis is combined, and makes it have preferable antitumor activity.
The content of the invention
The invention discloses a kind of compound represented containing following formula (I) or pharmaceutically acceptable salt or its prodrug
Wherein
A rings represent the macrocyclic polyamine compound that can be arbitrarily replaced;
L is a linking group, can be one kind in following several structures:
R1Independently selected from alkyl, haloalkyl;
R2Independently selected from one or more following groups:Substituted alkyl, substituted aryl, substituted heterocyclic aryl,
Halogen, hydroxyl, amino, alkylamino, dialkylamine, acid amides, cyano group, carboxyl, alkoxy carbonyl group, alkoxy, acyloxy ,=O ,=S,
=NH;
R3Independently selected from hydrogen, substituted alkyl, substituted aryl;
N is 0,1,2.
Formula (I) compound of the present invention, one of is preferably the compound with following formulas (II a):
Wherein
L is a linking group, can be one kind in following several structures:
R1Independently selected from methyl, ethyl, difluoromethyl, 2,2,2- trifluoroethyls;
R3Independently selected from hydrogen, substituted alkyl, substituted aryl;
R4, R5Independently selected from:Substituted alkyl, substituted aryl, substituted heterocyclic aryl, halogen, hydroxyl, amino, alkane
Amino, dialkylamine, acid amides, cyano group, carboxyl, alkoxy carbonyl group, alkoxy, acyloxy;
R6Independently selected from hydrogen, substituted alkyl, substituted aryl, halogen, hydroxyl, amino, alkylamino, dialkylamine,
Acid amides, cyano group, carboxyl, alkoxy carbonyl group, alkoxy, acyloxy ,=O ,=S ,=NH;
N is 0,1,2.
Formula (I) compound of the present invention, one of is preferably the compound with following formulas (II b):
Wherein
L is a linking group, can be one kind in following several structures:
R1Independently selected from methyl, ethyl, difluoromethyl, 2,2,2- trifluoroethyls;
R3Independently selected from hydrogen, substituted alkyl, substituted aryl;
R7, R8, R9Independently selected from:Substituted alkyl, substituted aryl, substituted heterocyclic aryl, halogen, hydroxyl, ammonia
Base, alkylamino, dialkylamine, acid amides, cyano group, carboxyl, alkoxy carbonyl group, alkoxy, acyloxy;
R10Independently selected from hydrogen, substituted alkyl, substituted aryl, halogen, hydroxyl, amino, alkylamino, dialkylamine,
Acid amides, cyano group, carboxyl, alkoxy carbonyl group, alkoxy, acyloxy ,=O ,=S ,=NH;
N is 0,1,2.
Formula (I) compound of the present invention, one of is preferably the compound with following formulas (II c):
Wherein
L is a linking group, can be one kind in following several structures:
R1Independently selected from methyl, ethyl, difluoromethyl, 2,2,2- trifluoroethyls;
R3Independently selected from hydrogen, substituted alkyl, substituted aryl;
R11, R12, R13Independently selected from:Substituted alkyl, substituted aryl, substituted heterocyclic aryl, halogen, hydroxyl, ammonia
Base, alkylamino, dialkylamine, acid amides, cyano group, carboxyl, alkoxy carbonyl group, alkoxy, acyloxy;
R14Independently selected from hydrogen, substituted alkyl, substituted aryl, halogen, hydroxyl, amino, alkylamino, dialkylamine,
Acid amides, cyano group, carboxyl, alkoxy carbonyl group, alkoxy, acyloxy ,=O ,=S ,=NH;
N is 0,1,2.
Formula (I) compound of the present invention, more preferably certainly:
Formula (I) compound of the present invention, its molecule can be coordinated with the salt containing metal ion.Its cationic can
To be main group metal ion or transition metal ions, anion can common are machine or inorganic anion.It is wherein described
Metal ion preferably be selected from Zn2+, Cu2+, Fe2+, Fe3+, Pt2+, Cr3+, Ni2+, Co2+, Mn2+, Ag+, Pd2+, La3+, Eu3+, Gd3+,
Ir3+, Tb4+, Lu3+, Ca2+;Described anion preferably is selected from F-, Cl-, Br-, I-, SO4 2-, SO3 2-NO3 -, PO4 3-, PO3 3-, ClO4 -,
ClO3 -, BrO4 -, IO4 -, oxalate denominationby, maleate ion, malate ion, citrate ion, succinic from
Son.
In addition, present invention also offers the method for synthesis type (I) compound.Raw material 5 and 6 can be according to Chinese patent CN
Method or the synthesis of similar method described in 101085743 and CN 101139358 are obtained.
If n is 0 in formula (I) compound, its synthetic method is:
If n is 1 or 2 in formula (I) compound, its specific building-up process can be illustrated by following reaction equation:
The reaction is reacted with CA-4 by the method that acyl chlorides or active ester is made with corresponding halogenated carboxylic acid, is obtained CA-4
Carboxylate.By carboxylate and cycleanine or derivatives thereof, synthesis obtains target product under base catalysis again.Finally this is produced
Thing obtains final product with metal ion complexation.
The compound and pharmaceutically acceptable salt of the present invention also includes the form of solvate or hydrate.It is general next
Say, the form of solvate or hydrate is equal with non-solvated or non-hydrated form, the model in the present invention is covered in the lump
In enclosing.Some compounds in the present invention there may exist polycrystal or unbodied form.Generally speaking, all physics shapes
Formula has equal purposes, and covers within the scope of the invention.
The present invention covers all stereoisomers of the compounds of this invention in form of mixtures or in pure form.This hair
The definition of bright compound includes all possible stereoisomer and its mixture.Its very specifically comprising racemic form and
The optical isomer with given activity of separation.Racemic form can be split by physical method, the physics side
Method such as carries out fractional crystallization, separation to diastereoisomer derivative or separated by chiral column chromatography.It can pass through
Conventional method such as optically active acid form then crystallizes into salt and obtains single optical isomer from racemic modification.
Present invention additionally comprises the prodrug of the compound.Prodrug is a kind of compound being derived by parent drug, its
Once entering internal, prodrug, which is just metabolized, is changed into parent drug.Prodrug can pass through one or more functions to parent drug
Group is replaced and prepared, and its substituted radical can be discharged parent compound by enzymatic in vivo.The preparation of prodrug and
Using can be in T.Higuchi and V.Stella, " Pro-drugs as Novel Delivery System ", Vol.14of
The A.C.S.Symposium Serier and Bioreversible Carriers in Drug Design, ed.Edward
B.Roche, AmericanPharmaceutical Association and Pergamon Press, find in 1987.
The invention further relates to (I) compound of the formula comprising the effective dose and pharmacologically pharmaceutical composition of acceptable carrier,
Said composition is applied to local, enteral or parenteral administration, can be it is inorganic or organic, it is solid-state or liquid.
For oral, especially with tablet or capsule.This tablet or capsule include active component and diluent (such as lactose, glucose, sugarcane
Sugar, mannitol, sorbierite, cellulose, glycerine), lubricant (such as talcum, stearate), polyethylene glycol.Tablet can also be wrapped
Containing adhesive, starch, gelatin, methylcellulose, sodium carboxymethylcellulose and/or polyvinylpyrrolidone, it can also wrap if necessary
Containing disintegrator (such as starch, agar, alginic acid and its salt), effervescent mixture, or adsorbent, dyestuff, flavor enhancement, sweetener.These
Composition can be applicable in the form of parenteral or in the form of injection.The such preferred isotonic aqueous solution of formulation or breast
Liquid, such as in the case of the freeze-dried composition being only made up of active component and a kind of carrier (such as mannitol), such solution can be with
Using preceding preparation.These pharmaceutical compositions can be sterile, or comprising excipient, or solubilizer, regulation osmotic pressure
Salt.
The medicine of the treatment disease related to tumour is being prepared the invention provides a kind of medicine comprising formula (I) compound
In application.Tumour therein includes:Lung cancer, ED-SCLC, liver cancer, cancer of pancreas, stomach cancer, osteocarcinoma, cancer of the esophagus, mastocarcinoma,
Kidney, cholangiocarcinoma, prostate cancer, carcinoma of testis, colon cancer, oophoroma, carcinoma of urinary bladder, cervix cancer, bronchiolar carcinoma, melanoma,
Gland cancer, syringocarcinoma, papillary carcinoma, papillary adenocarcinoma, squamous cell carcinoma, basal-cell carcinoma, adenocarcinoma cystic, spongiocytoma, star
Type cytoma, medulloblastoma, neuroblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma,
Oligodendroglioma, meningioma, neurofibroma, fibrosarcoma, desmocytoma, fibroma, myxosarcoma, mucous bursa
Knurl, lipoma, lipoadenoma, chondrosarcoma, chondroma, chondromyoma, chordoma, chorioadenoma, villous vessels knurl, fine hair
Epithelioma, chorioblastoma, osteosarcoma, osteoblastoma, osteoclastoma, osteochondrofibroma, osteochondrosarcoma, stock
Cystoma, osteodentinoma, osteofibroma, the fibrosarcoma of bone, hemangioma, angiosarcoma, lymphangioendothelial sarcoma, lymphangioma, lymthoma,
Endothelioma, synovialoma, synovial sarcoma, celiothelioma, mesocytoma, ewing's tumor, liomyoma, leiomyosarcoma, rhabdomyoma,
Rhabdomyosarcoma, acute lymphatic leukemia, acute myelogenous leukemia, chronic leukemia, polycythemia, multiple bone
Myeloma.
Present invention also offers a kind of test method for being used to treating or suppressing tumour, this method is mainly by Mice Body
Experiment, determines the antitumor activity of compound.
Embodiment
In order to be best understood from the present invention, present disclosure is expanded on further with reference to embodiment, but the present invention's is interior
Appearance is not limited solely to the following examples.
The following is the definition for the term that can be used in this manual.Unless otherwise indicated, this patent is with regard to group or term
For the original definition that provides suitable for specification in the whole text the group or term, no matter being single use or making
Part for another group is used.
Term " alkyl " refers to straight chain or side chain unsubstituted alkyl, and it has 1-20 carbon atom, preferably
1-6 carbon atom, refers in particular to methyl, ethyl, propyl group (including n-propyl and isopropyl), butyl (including normal-butyl, isobutyl
Base, the tert-butyl group) etc..
Term " halogen " or " halo " refer to fluorine (fluoro), chlorine (chloro), bromine (bromo), iodine (iodo).
Term " aryl " refers to monocyclic or polycyclic aromatic hydrocarbons, such as benzene, naphthalene, anthracene, phenanthrene.
Term " heterocyclic aryl " refers to optionally substituted aromatic cyclic groups, and wherein at least contains a carbon atom by it
Its hetero atom replaces, and hetero atom includes nitrogen, oxygen, sulphur.The nitrogen and sulfur heteroatom also can be optionally oxidized, and nitrogen heteroatom also can be optional
It is quaternized.The heterocyclic group can be connected at any hetero atom or carbon atom.It is preferred that heterocyclic aryl include but is not limited to, pyrrole
It is pyridine, pyrazine, pyrimidine, pyridazine, triazine, furans, thiophene, imidazoles, triazole, tetrazolium, thiazole, isothiazole, pyrroles, pyrazoles, oxazoles, different
Oxazole, benzofuran, benzothiazole, benzothiophene, indoles, quinoline, isoquinolin, purine, carbazole, benzimidazole, pyrrolo- pyrrole
Pyridine, pyrrolopyrimidine etc..
Term " cycloalkyl " refers to the carbocyclic ring of non-aromatic, including monocyclic, condensed ring or loop coil.Cycloalkyl also includes having one
The ring of individual or multiple aromatic rings fusions (having a common key), the cycloalkyl for having one or more aromatic rings fusions can lead to
Cross aromatic rings or non-aromatic ring part is connected with other groups.
Term " Heterocyclylalkyl " refers to nonaromatic heterocycles, and wherein one or more ring member nitrogen atoms are hetero atoms, such as oxygen, nitrogen,
Sulphur atom.Heterocyclylalkyl can include monocyclic or polycyclic (if any 2,3,4 fused rings), loop coil.It is preferred that Heterocyclylalkyl include
Aziridine, azetidine, tetrahydrofuran, thiophane, pyrrolidines, oxazolidines, thiazolidine, isothiazolidine, imidazolidine, pyrazoles
Alkane, morpholine, thiomorpholine, piperazine, piperidines etc..Heterocyclylalkyl also includes the heterocycle condensed with one or more aromatic rings, example
Such as 2,3- Dihydrobenzofuranes, 1,3- benzodioxolanes, phendioxin, 4- dioxanes, benzenedicarboxamide etc..With one or many
The Heterocyclylalkyl of individual aromatic rings fusion can be connected by aromatic rings or non-aromatic ring part with other groups.
Term " amide groups " refers to group-C (=O) NH-.
Term " cyano group " refers to group-CN.
Term " alkylamino " refers to by an alkyl-substituted amino.
Term " dialkylamine " refers to group by two identical or different alkyl-substituted amino.
Term " carboxyl " refers to group-COOH.
Term " alkoxycarbonyl " refers to group-C (=O) OR15, wherein R15Refer to alkyl.
Term " alkoxy " refers to group-OR16, wherein R16Refer to alkyl.
Term " acyloxy " refers to group-OC (=O) R17, wherein R17Refer to alkyl.
" substituted " mean the group that then describes can by some common groups (such as hydrogen, halogen, hydroxyl, amino,
Sulfydryl, nitro, cyano group, aryl, heterocyclic radical, Heterocyclylalkyl, carboxyl, amide groups etc.) substitution.
" optional " means that the event then described or situation can occur or not occur, and the description is paraded one's wealth wherein
The example that the event or situation occur and the example that wherein it does not occur.
" pharmaceutically acceptable carrier " used herein includes any and whole solvent, decentralized medium, coating, antibacterium
With antifungal medicine, etc. blend absorption delaying agent etc..It is many in this area that such medium and medicament, which are used for pharmaceutically active substances,
Well known., can be pre- during its application in therapeutic combination unless any conventional media or medicament are incompatible with active component
Phase.The active component of supplement also may be incorporated into composition.
Embodiment 1
Step 1:
(Z) -1- (3,4,5- trimethoxyphenyls) -2- (3- amino-4-methoxyls phenyl) is added in 500ml there-necked flasks
2- chloracetyl chlorides are added under ethene (31.5g, 0.1mol), dichloromethane 300ml, diisopropylethylamine 20ml, stirring, ice bath
(11.3g, 0.1mol), is warming up to 25 DEG C after adding, react 3h, TLC tracking.It is water-soluble with sodium acid carbonate respectively after reaction terminates
Liquid, saturated common salt water washing, collect organic phase, and anhydrous magnesium sulfate is dried, and is filtered, concentration.Obtained crude product is directly used in lower step
Reaction.
Step 2:
The crude product that upper step is obtained is dissolved in 300ml DMF, adds PdCl2(17.7g, 0.1mol), tetrabutylammonium chloride
(27.8g, 0.1mol), cycleanine (17.2g, 0.1mol), stirring is heated to 80 DEG C, reacts 10h, TLC tracking.Reaction knot
Shu Hou, cooling.Reaction solution is poured into 500ml frozen water, extracted 3 times with 300ml dichloromethane respectively, organic phase is collected, it is anhydrous
Magnesium sulfate is dried, filtering, is concentrated to give crude product.Crude product is separated with flash column chromatography, eluant dichloromethane/methanol/triethylamine
=9/1/0.5, obtain sterling 25g, yield 47.4%.MS (M+1)=528.6.
Embodiment 2
Step 1:The preparation of Cyclen -1,4,7,10- tetraacethyls (DOTA)
Isosorbide-5-Nitrae is added in there-necked flask, 7,10- tetraazacyclododecanands (100g, 0.58mol), deionized water 1000ml is stirred
Mix, 30% KOH solution be added dropwise, then regulation pH value adds monoxone (263g, 2.78mol) to 8.5, then with 30% KOH
Solution adjusts pH value to 8.5, is heated to 80 DEG C, reacts 24h, and pH value is maintained in the process between 8.5-9.Reaction terminates
Afterwards, cool down, add concentrated hydrochloric acid and adjust pH value to 2, have white precipitate generation, filter.Filter cake is recrystallized with water-ethanol solution, is obtained
The crystal ethanol arrived, ether washing, dries, obtains DOTA crystal 183g, yield 78%.MS (M+1)=405.4.
Step 2:
DOTA (20g, 0.05mol), thionyl chloride 200ml are added in the there-necked flask that band is stirred at reflux device, is stirred, plus
Heat backflow 4h, decompression boils off thionyl chloride liquid after cooling, adds the THF of 100ml dryings, and decompression boils off solvent, consolidate
Body acyl chlorides.This obtained solid acid chloride is dissolved in the dichloromethane of 250ml dryings, diisopropylethylamine 20ml, stirring, ice is added
Bath is lower to add (Z) -1- (3,4,5- trimethoxyphenyl) -2- (3- hydroxyl -4- methoxyphenyls) ethene (4g, 0.0125mol),
Ice bath is removed, 30 DEG C are to slowly warm up to, 3h, TLC tracking is reacted.Reaction adds frozen water 100ml after terminating, and stirs 30min, decompression
Boil off organic solvent.In aqueous phase with concentrated hydrochloric acid adjust pH value to 2, have solid precipitation, be filtrated to get white solid, with ethanol-
The aqueous solution is recrystallized, and obtains target product 22.8g, yield 65%.MS (M+1)=703.8.
Embodiment 3
(Z) -1- (3,4,5- trimethoxyphenyl) -2- (3- amino-4-methoxyls phenyl) ethene (20g, 63mmol) is molten
In 200ml dichloromethane, add and add 1 under diisopropylethylamine 13ml, stirring, ice bath, 1 '-carbonyl dimidazoles (12.3g,
76mmol), DMF (100ml) solution of cycleanine (13g, 76mmol) is added after reaction 1.5h, was reacted at room temperature after adding
Night, TLC tracking.After reaction terminates, reaction solution is concentrated to dryness, separated with flash column chromatography, separation condition is eluant, eluent:Dichloro
Methane/methanol/triethylamine=9/1/0.5, obtains target product 23g, yield 71%.MS (M+1)=514.6.
Embodiment 4
Isosorbide-5-Nitrae, 7,10- tetraazacyclododecanands (20g, 0.116mol) are dissolved in 200mlDMF, add N, N- dimethyl methyls
Acid amides contracting dicarbaldehyde (20.8g, 0.17mol), stirring is heated to 100 DEG C, reacts 2h, after reaction terminates, is cooled to room temperature.Will
Reaction solution is poured into 500ml water, and stood overnight at room temperature.Then extracted 3 times, each 200ml, merged organic with chloroform
Layer, anhydrous magnesium sulfate is dried, and decompression boils off solvent, that is, obtains 1- formaldehyde-Isosorbide-5-Nitrae, 7,10- teteaazacyclododecanes.
Solid obtained in the previous step is dissolved in 350ml dichloromethane, iodomethane (49.5g, 0.35mol), hydrogen-oxygen is added
Change potassium 20g, be heated to 50 DEG C, stirring reaction 3h.After the completion of reaction, cool down, filtering is concentrated the filtrate to dry.By consolidating for obtaining
After body is dissolved with 200ml ethanol, 10% sulfuric acid solution 100ml is added, 50 DEG C, stirring reaction 5h are heated to.After reaction terminates,
Cooling, pH value is adjusted to 9.5 with 10% potassium hydroxide, is added dichloromethane and is extracted 3 times, merges organic layer, is concentrated after drying
It is extremely dry.Flash column chromatography is separated, and separation condition is eluant, eluent:Methylene chloride/methanol/triethylamine=9/1/0.5, obtains 4,7,
10- trimethyls-Isosorbide-5-Nitrae, 7,10- tetraazacyclododecanand 16.7g, yield 60%.MS (M+1)=215.4.
(Z) -1- (3,4,5- trimethoxyphenyls) -2- (3- amino-4-methoxyls phenyl) ethene is added in there-necked flask
2- chloracetyl chlorides are added under (31.5g, 0.1mol), dichloromethane 300ml, diisopropylethylamine 20ml, stirring, ice bath
(11.3g, 0.1mol), is warming up to 25 DEG C after adding, react 3h, TLC tracking.It is water-soluble with sodium acid carbonate respectively after reaction terminates
Liquid, saturated common salt water washing, collect organic phase, and anhydrous magnesium sulfate is dried, and is filtered, concentration.Obtained crude product is directly used in lower step
Reaction.
The crude product that upper step is obtained is dissolved in 300ml DMF, adds PdCl2(17.7g, 0.1mol), tetrabutylammonium chloride
(27.8g, 0.1mol), 4,7,10- trimethyls-Isosorbide-5-Nitrae, 7,10- tetraazacyclododecanands (21.4g, 0.1mol) are stirred, heating
To 80 DEG C, 10h, TLC tracking are reacted.After reaction terminates, cooling.Reaction solution is poured into 500ml frozen water, respectively with 300ml bis-
Chloromethanes is extracted 3 times, collects organic phase, and anhydrous magnesium sulfate is dried, and filtering is concentrated to give crude product.Crude product flash column chromatography point
From eluant dichloromethane/methanol/triethylamine=9/1/0.5 obtains sterling 25g, yield 47.4%.MS (M+1)=570.7.
The similar method more than, this patent is also prepared for following compound:
The preparation of zinc salt complex
1mmol compounds are dissolved in 100mL absolute ethyl alcohols, the nothing containing 1mmolZn (ClO4) 26H2O is slowly added dropwise
Hydrous ethanol solution, is stirred at room temperature 2h, and filtering, vacuum drying respectively obtains following several compounds:
The preparation of platinum salt complex
1mmol compounds are dissolved in 100mL absolute ethyl alcohols, are slowly added dropwise containing addition K2PtCl4 (0.42g, 1mmol)
Temperature is risen into 50 DEG C of lucifuges after the aqueous solution (5mL) and reacts 12h.Reaction solution adds acetone (20mL) precipitation after being concentrated under reduced pressure dark blue
Color precipitate, precipitation washed with a small amount of acetone 3 times, then it is vacuum dried after respectively obtain following several compounds:
The pharmaceutical formulation of embodiment 60
The invention provides several formulas for being used to treat or prevent the pharmaceutical composition of the disease related to protein kinase,
Its pharmaceutical composition has tablet, capsule, injection, aerosol etc..The change shown in the present invention is represented with " reactive compound " below
Compound.
Injection (a) |
(50mg/ml) |
Active material |
5.0%w/v |
1M sodium chloride solutions |
15.0%w/v |
0.1M hydrochloric acid solutions adjust pH to 7.6 |
|
PEG400 |
4.5%w/v |
Add water to 100% |
|
Injection (b) |
(10mg/ml) cushioning liquid (pH 6) |
Active material |
1.0%w/v |
Sodium phosphate |
2.26%w/v |
Citric acid |
0.38%w/v |
PEG400 |
3.5%w/v |
Add water to 100% |
|
The efficacy experiment of the Nude Mice of embodiment 61
BALB/cA-nude nude mouses, 6-7 weeks, ♀, purchased from Shanghai Slac Experimental Animal Co., Ltd..Nude mouse
Human hepatocarcinoma Bel-7402 cell, colon cancer cell line HT-29, SGC-7901 cell and non-small cell lung cancer are subcutaneously inoculated with respectively
A549 cells, treat tumour growth to 100-250mm3Afterwards, animal is grouped to (d0) at random.2-3 knurl volume is surveyed weekly, claims mouse
Weight, record data.Gross tumor volume (V) calculation formula is:
V=1/2 × a × b2 wherein a, b represent length and width respectively;
T/C (%)=(T-T0)/(C-C0) × 100 wherein T, C is the gross tumor volume at the end of experiment;
Gross tumor volume when T0, C0 start for experiment.
Institute's test agent is used to 50%PEG400 distilled water dilutings into required concentration.Application method is is administered once daily, abdomen
Chamber is injected, and is used in conjunction 21 days.Respectively to compound shown in embodiment 1,3,4,34,36,37,47,49,50 to human liver cancer Bel-
7402nd, the curative effect of the Nude Mice of SGC-7901 cell and Non-small Cell Lung Cancer A 549 is tested, its
As a result it is as shown in table 2.
The curative effect of a variety of cancer cell Nude Mices of the reactive compound of table 2
Conclusion:Compound shown in of the invention significantly inhibits human liver cancer Bel-7402, stomach cancer SGC-7901 under test conditions
The growth of cell and Non-small Cell Lung Cancer A 549 Nude Mice, especially its metal complex have more obvious suppression
The effect of tumour cell processed.