CN105601903B - A kind of high-molecular compound with active anticancer, its preparation method and application - Google Patents
A kind of high-molecular compound with active anticancer, its preparation method and application Download PDFInfo
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- CN105601903B CN105601903B CN201510996205.4A CN201510996205A CN105601903B CN 105601903 B CN105601903 B CN 105601903B CN 201510996205 A CN201510996205 A CN 201510996205A CN 105601903 B CN105601903 B CN 105601903B
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- anticarcinogen
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- C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
- C08G2650/00—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule
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Abstract
The invention belongs to macromolecule coupling anticarcinogen technical field, specifically related to a kind of high-molecular compound PEG (A+G) with active anticancer, its preparation method and application, the present invention makes anticarcinogen reach cancer focus with Chk1 inhibitor " while and by a certain percentage " via ethylene glycol copolymer nano target carrier, reduce the toxicity of anticarcinogen, improve the curative effect of anticarcinogen, can solve anticarcinogen and Chk1 inhibitor clinically associated with problem, experiment is proved, above-mentioned PEG (A+G) has to the Nude Mices of Colo 205 significantly inhibits effect, there is application prospect in terms of PTS is prepared.
Description
Technical field
The invention belongs to macromolecule coupling anticarcinogen technical field, and in particular to it is a kind of with active anticancer by poly- second two
High-molecular compound, its preparation method and the application of alcohol nano target carrier conjugation anticarcinogen and Chk1 inhibitor.
Background technology
Chemotherapy is the important means for the treatment of cancer, but traditional chemotherapy is present that toxic side effect is big, the low problem of curative effect.Previous generation
Record latter stage, as Trouet and De Duve have found that cell endocytic approach can be used for drug delivery to lysosome, Ringsdorf
Propose macromolecule coupling drug model and Maeda is found that " enhancing infiltration and reservation " (EPR) effect, nanometer technology is in health
Gradually it is used widely in science, has expedited the emergence of nanosecond medical science this new disciplines, main research object is Nano medication transmission
System (NDDS), including lipidosome cream, nanoparticle suspension thing, biopolymer nanoparticles, macromolecule are treated and for gene delivery
The direction such as nano particle, the bioavilability and pharmacokinetics of medicine can be significantly increased, with high medicinal
With commercial value.Wherein, macromolecule treatment is nanosecond medical science field the most successful, the past two during the last ten years, in this field
There are 11 macromolecule coupling drugs to enter market sale by FDA approvals, wherein 10 with polyethylene glycol (PEG) or polycyclic oxygen second
Alkane (PEO) is carrier, and 5 are that macromolecule is coupled anticarcinogen, and also multiple macromolecule coupling anticarcinogens enter clinical test in addition.
Substantial amounts of preclinical test shows with clinical test：Anticancer can be greatly reduced in macromolecule coupling anticarcinogen nano target medicine
The toxicity of medicine, significantly improves its curative effect, is the breakthrough to traditional medicine, it has also become the new direction of medicament research and development.
Have much as the research of medicine macromolecule carrier about polyethylene glycol.Relatively other macromolecule carrier such as N-
For (2- hydroxypropyls) methacrylamide copolymer, polyglutamic acid etc., polyethylene glycol carrier is by U.S.'s food Drug
Office (FDA) approval progress is commercially use, has generally acknowledged safety indexes, has achieved huge success.Zhao et al.
Once single armed polyethylene glycol conjugation daunorubicin, four arm polyethylene glycol conjugation SN-38 and bifurcated polyethylene glycol conjugation four were synthesized poly-
Or eight poly- cytarabines etc., solubility, half-life period and the bioavilability of medicine has all been significantly increased, medicine is significantly reduced
The toxicity of thing, improves the anticancer live body curative effect of medicine.
Most of treatments of cancer are realized by induced DNA damage with killing cancer cell.By anticarcinogen chemotherapy or
Under gene poison stress situation caused by radiation therapy, in order to maintain the integrality of gene and the existence of cell, include at one thin
The complex network of the multiple signal path of born of the same parents' circular test point, DNA reparations, transducer and cell suicide etc. there occurs so-called
" DNA damage reaction " (DDR), causes cell cycle to stagnate temporarily so that cell has the damage of time DNA plerosis.Chk1
(Checkpoint Kinase 1:Checkpoint kinases 1) it is the core ingredient that DNA damage reacts (DDR), suppressing Chk1 can be special
Surely function of the DNA damage reagent to the selectively killing of the defective cancer cells of p53 is strengthened.
In the normal cell of mammal, stagnated as caused by tumor suppressor p 53 regulation G1 caused by DNA damage, because
And normal cell is mainly stagnated in G1 under the malicious stress situation of gene；Most of tumour cells have p53 and RB pathway deficiencies, and
The defective tumour cells of p53 are defective in G1 checkpoints, stagnate and depend critically upon S or G2 checkpoints to repair damage
DNA and the integrality that gene is kept for basic existence, thus, in the special circumstances for suppressing Chk1, eliminating S and G2 checkpoints
Under, normal cell remains to be stuck in G1 to be damaged with DNA plerosis；, then will be through having silk point all through the ages without the tumour cell of G1 checkpoints
Disaster is split, so as to cause cell suicide.Obviously, tumor cell ratio normal cell relys more on Chk1 kinases to maintain cell cycle
Stagnate, therefore, specifically selection of the enhanced sensitivity DNA damage reagent (chemotherapy or radiation therapy) to cancer cell can be used to by suppressing Chk1
Property kill.In a variety of cancers, based on p53 approach, it can set up treatment window to increase with one or more Chk1 inhibitor
Strong treatment of cancer.Therefore, research and development Chk1 inhibitor can as treating cancer new way.
Existing many companies have carried out substantial amounts of research work to Chk1 inhibitor.Abbott Laboratories are synthesized
Urea series compound such as A-690002 etc. is as Chk1 inhibitor, and Pre-clinical animal studies show such compound significantly
Ground enhances the curative effect of the anticarcinogens such as adriamycin, camptothecine, Irinotecan, and toxicity is faint.And AstraZeneca and Pfizer
The scientist of company has then synthesized AZD7762 and PF-477736 etc., is replaced respectively with gemcitabine, Yi Li in preclinical test
The various anticarcinogen combinations such as health, Docetaxel, carboplatin, significantly enhance live body curative effect.But so far, there is not yet
The report that Chk1 inhibitor phase iii clinical trials succeed, reason is probably many, such as anticarcinogen and Chk1 inhibitor
Toxic side effect, anticarcinogen and Chk1 inhibitor be difficult to " while and by a certain percentage " and reach cancer location to play drug effect etc..
The content of the invention
To solve the problems, such as that the technical scheme that above-mentioned clinical test is used is：
By anticarcinogen and Chk1 inhibitor by amino acid connects chain graft on simultaneously on polyethylene glycol nano-carrier with
Synthesizing polyethylene glycol coupling (anticarcinogen with Chk1 inhibitor), obtain a class make anticarcinogen and Chk1 inhibitor be integrated into one it is whole
The nano target medicine of body, reaches the toxic side effect that anticarcinogen and Chk1 inhibitor is greatly reduced and significantly improves its curative effect
Effect, it is true to be formed while anticarcinogen and Chk1 inhibitor " while and by a certain percentage " can be made to reach cancer location again
" therapeutic alliance " in positive meaning, solve anticarcinogen and Chk1 inhibitor clinically associated with problem so that live body curative effect is again
Significantly improve, so as to start a class PTS.
High-molecular compound of the present invention with active anticancer, its structural formula is as shown in XI：
Invention additionally discloses above-mentioned high-molecular compound XI preparation method, it is characterised in that：Comprise the following steps,
Under HBTU, HOBT, DIEA and solvent DMF, 0 DEG C to room temperature condition, by Boc-Gly and Chk1 inhibitor
AZD7762 carries out amidatioon connection, is gone through TFA after Boc protects, and the glutamic acid that the Gly-AZD7762 and Fmoc of gained are protected-
The 5- tert-butyl esters 4,6- trimethylpyridines, carry out amidatioon connection under the conditions of 0 DEG C in PyAOP and 2, are then gone with triethylamine and DMF
Fmoc is protected, then the 6-aminocaprolc acid protected with Fmoc is in PyAOP and 2,4,6- trimethylpyridines, carries out acid amides under the conditions of 0 DEG C
Change connection, then go Fmoc to protect with triethylamine and DMF conditions again, gained micromolecular compound intermediate and quadrifurcate poly- second
Glycol macromolecule carrier (4ARM-SCM-40K) carries out substituted amide connection, and gained high-molecular compound carries out uncle through TFA again
Butyl ester is deprotected, and the hydroxy-acid group of gained aromatic polymer intermediates is again through n-hydroxysuccinimide, DCC, DMAP and CH2Cl2Condition
Activated, be finally coupled under the conditions of pyridine with gemcitabine, obtain final products XI.
The invention also discloses applications of the above-mentioned high-molecular compound XI in anticarcinogen is prepared.According to anti-human Colon and rectum
Cancerous cell line Colo-205 embodiment experimental result shows, described high-molecular compound XI dosage for 700~
During 800mg/kg, experimental endpoints Relative tumor proliferation rate T/C (%) is 60% or so, can show significant antitumor work
With.
Brief description of the drawings
Fig. 1：Midbody compound VII proton nmr spectra (1H-NMR)。
Fig. 2：Intermediate high-molecular compound VIII proton nmr spectra (1H-NMR)。
Fig. 3：Intermediate high-molecular compound IX proton nmr spectra (1H-NMR)。
Fig. 4：PEG (A+G) (XI) proton nmr spectra (1H-NMR)。
Fig. 5：PEG (A+G) (XI), small-molecule drug gemcitabine and the combination of Chk1 inhibitor and the tumour life of physiological saline
Long curve.
Embodiment
Following non-limiting examples can make one of ordinary skill in the art be more fully understood the present invention, but not with
Any mode limits the present invention.
Embodiment 1
Ethylene glycol copolymer coupling (anticarcinogen gemcitabine and Chk1 inhibitor AZD7762) (for convenience of stating, is write a Chinese character in simplified form
For PEG (A+G)) synthesis
By Boc-Gly (265.8mg, 1.5176mmol, self-control), AZD7662 (500mg, 1.3796mmol, the auspicious poem in Shanghai
Chemical Co., Ltd.), HBTU (776.6mg, 2.0694mmol), HOBT (279.6mg, 2.0694mmol) is added to 250mL's
In flask, and dissolved with DMF (15mL), stirring cooling 20 minutes, are then slowly added dropwise under the conditions of mixed solution is transferred into 0 DEG C
DIEA (1.08mL, 6.2082mmol).Move to and be stirred at room temperature after 2 hours, overnight.Reaction stops, and reaction solution is poured into saturation
NaHCO3In solution (200mL), (100mL × 4) are extracted with ethyl acetate, saturation NaHCO is then used3Solution (150mL) cleaning has
Machine phase, then cleaned with saturation NaCl solution (100mL).The anhydrous MgSO of final organic phase4Dry, then filtering depressurizes dense
Solution is evaporated by contracting.Silica gel column chromatography, with 5% MeOH/CH2Cl2Elution, collects product, and evaporated under reduced pressure obtains product (for convenience
Statement, is abbreviated as compound I) 530.7mg, yield 74.1%.1H-NMR(500MHz,DMSO-d6)δ10.05-9.55(m,1H),
8.45-8.22(m,1H),8.15-7.98(m,1H),7.65-7.38(m,1H),7.34-7.18(m,1H),6.95-6.28(m,
3H),4.45-4.12(m,1H),3.90-3.68(m,3H),3.65-3.55(m,1H),3.05-2.92(m,1H),2.65-2.52
(m,1H),1.98-1.84(m,1H),1.82-1.56(m,2H),1.54-1.28(m,10H)；ITMS+c ESI：[M+Na+]
542.2173
Take compound I (500mg, 0.9630mmol) to be placed in 25mL round-bottomed flasks, add dichloromethane (5mL), then delay
It is slow that TFA (0.074mL, 9.630mmol) is added dropwise, it is stirred overnight at room temperature.It is spin-dried for, crude product methanol dissolves, adds bicarbonate
Sodium (211.2mg, 2.5137mmol) neutralizes unnecessary TFA.Solid impurity is filtered to remove, silica white (3g) is added into filtrate,
It is spin-dried for that solid solution is made, dry method loading crosses silicagel column purifying.First use 5%MeOH/CH2Cl2Elution, then use 4%NH3·H2O/
8%MeOH/CH2Cl2To 5%NH3·H2O/10%MeOH/CH2Cl2Gradient elution.Collect product, be concentrated and dried product (is
Convenient statement, is abbreviated as intermediate product II) 403.5mg, yield 100%.1H-NMR(500MHz,DMSO-d6)δ10.05-9.8
(m,1H),8.32-8.24(m,1H),8.22-8.02(m,1H),7.55-7.44(m,3H),7.28-7.22(m,1H),6.84-
6.48(m,2H),4.44-4.10(m,1H),4.01-3.68(m,3H),3.65-3.58(m,1H),3.15-2.90(m,1H),
2.82-2.60(m,1H),1.98-1.84(m,1H),1.84-1.75(m,1H),1.74-1.58(m,1H),1.56-1.35(m,
1H)；MALDI-TOF MS:[M+Na+]442.1302
By intermediate product II (403.7mg, 0.9630mmol), PyAOP (702.9mg, 1.3482mmol), Fmoc-L-
Glu-OH (5-Bu-t) (573.6mg, 1.3482mmol) is added in 250mL flask, and is dissolved with DMF (15mL), will be mixed
Solution be transferred to 0 DEG C under the conditions of stirring cooling 20min, be then slowly added dropwise 2,4,6- trimethylpyridines (0.127mL,
0.9630mmol).Stirred under the conditions of moving to 3 DEG C after reaction 4h, overnight.Reaction solution is poured into saturation NaHCO3(200mL) solution
In, it is extracted with ethyl acetate (100mL × 10).Then saturation NaHCO is used3Solution (150mL) is eluted 1 time, molten with saturation NaCl
Liquid (100mL) is eluted 1 time.Merge organic phase, be then concentrated under reduced pressure and be evaporated liquid, dissolved, added with ethyl acetate and methanol
Proper silica gel powder, dry method loading.Silica gel column chromatography, first uses 200ml CH2Cl2Elution, then use 0.5%MeOH/CH2Cl2To 2%
MeOH/CH2Cl2Elution, finally uses 5%NH3·H2O/10%MeOH/CH2Cl2Elution.Collect product be evaporated product (for convenience
Statement, is abbreviated as intermediate product III) 558.9mg (70.1%).1H-NMR(500MHz,DMSO-d6)δ10.20-9.90(m,
1H),8.35-8.25(m,1H),8.08-8.02(m,1H),7.98-7.94(m,2H),7.92-7.88(m,2H),7.80-7.70
(m,2H),7.65-7.58(m,1H),7.55-7.40(m,5H),7.38-7.32(m,2H),7.28-7.25(m,1H),6.78-
6.60(m,1H),4.30-4.28(m,1H),4.26-4.20(m,3H),4.03-3.96(m,1H),3.94-3.88(m,1H),
3.86-3.70(m,2H),2.98-2.92(m,1H),2.70-2.55(m,1H),2.40-2.22(m,2H),2.05-1.85(m,
2H),1.70-1.60(m,2H),1.52-1.30(m,11H)；MALDI-TOF MS:[M+Na+]849.2960,[M+K+]
865.2725
By intermediate product III (550.0mg, 0.6764mmol), triethylamine (68mg, 6.764mmol) is added to 150mL's
In flask, dissolved with DMF (10mL), react 3h, stop reaction.Reaction solution is poured into saturation NaHCO3In (200mL) solution, use
Ethyl acetate extracts (100mL × 4).Then saturation NaHCO is used3Solution (150mL) is eluted 1 time, uses saturation NaCl solution
(100mL) is eluted 2 times.Dried with anhydrous MgSO4, suction filtration, addition (3g) silica white in filtrate, dry method loading, silica gel column chromatography,
First use 100ml CH2Cl2Elution, then use 2%MeOH/CH2Cl2To 6%MeOH/CH2Cl2Elution, finally uses 5%NH3·H2O/
10%MeOH/CH2Cl2Elution.Collect product, be evaporated to obtain IV intermediate product 312.8mg, yield 76.7%.1H-NMR (500MHz,
DMSO-d6)δ10.05-9.95(m,1H),8.40-8.22(m,1H),8.15-7.82(m,2H),7.62-7.36(m,3H),
7.35-7.20(m,1H),6.90-6.45(m,2H),4.45-4.26(m,1H),4.25-4.10(m,1H),4.00-3.90(m,
2H),3.88-3.64(m,2H),3.25-3.15(m,1H),2.98-2.90(m,1H),2.72-2.55(m,1H),2.35-2.22
(m,2H),1.98-1.80(m,2H),1.68-1.55(m,2H),1.51-1.49(m,1H),1.48-1.40(m,9H)；MALDI-
TOF-MS:[M+H+] 605.3076, [M+Na+] 627.2876, [M+K+]643.2742
6-aminocaprolc acid (2.3047g, 17.57mmol) is added in 500mL flask, 150mL (75/ is then added
75) THF/H2O, mixed solution is stirred at 0 DEG C cooling 20 minutes, adds Na2CO3Solid (3.2475g,
35.14mmol).It is added dropwise after 20 minutes after Fmoc-Cl (5.0g, 19.327mmol) THF solution (10mL) is dripped off and moves to room temperature
1h is stirred, with 10% lemon acid for adjusting pH value to 3.Reaction solution is transferred in 1L separatory funnel, is extracted with ethyl acetate
(100mL × 4 time), (120mL × 2) are eluted with 1N HCl.Merge organic phase, use anhydrous MgSO4Dry, then suction filtration depressurizes
Liquid is evaporated by concentration, is dissolved with 5mL ethyl acetate, n-hexane (300mL) sedimentation is added while stirring, crude product is with powdered analysis
Go out.Crude product ethyl acetate is dissolved, proper silica gel powder is added, makes solid solution, dry method loading, column chromatography uses 20% second
Acetoacetic ester/petroleum ether to 80% ethyl acetate/petroleum ether carries out gradient elution.Product is collected, is concentrated under reduced pressure and is evaporated to obtain middle production
Thing V 6.209g, yield 99.0%.1H-NMR (500MHz, DMSO-d6)δ12.5-11.75(m,1H),7.99-7.80(m,2H),
7.78-7.60(m,2H),7.49-7.38(m,2H),7.37-7.29(m,2H),7.28-7.18(m,1H),4.45-4.20(m,
3H),2.20-2.15(m,2H),2.10-2.00(m,2H),1.50-1.40(m,3H),1.40-1.35(m,3H)
By intermediate product IV (550mg, 0.9095mmol), intermediate product V (482.2mg, 1.3643mmol), EDCI
(261.53mg, 1.3643mmol), HOBT (221.2mg, 1.6371mmol) is added in 250mL flask, uses CH2Cl2
(15mL) is dissolved, and solution stir under the conditions of 0 DEG C and cooled down 20 minutes, be then slowly added dropwise DIEA (0.285mL,
1.6371mmol).Move to and be stirred at room temperature after reaction 4h, overnight, stop reaction, be directly evaporated.By crude product CH2Cl2Dissolving, it is wet
Method loading, column chromatography first uses 200ml CH2Cl2Elution, then use 2%MeOH/CH2Cl2To 5%MeOH/CH2Cl2Elution, finally
Use 2%NH3·H2O/4%MeOH/CH2Cl2Elution.Collect product be evaporated product (for convenience of state, be abbreviated as intermediate product
VI) 855.2mg, yield 100%.1H-NMR (500MHz, DMSO-d6) δ 10.00-9.78 (m, 1H), 8.88-8.54 (m, 1H),
8.28-8.16(m,1H),7.98-7.86(m,2H),7.84-7.74(m,3H),7.65-7.56(m,2H),7.48-7.31(m,
3H),7.30-7.28(m,2H),7.28-7.22(m,2H),7.18-7.12(m,2H),6.68-6.51(m,1H),4.35-4.22
(m,1H),4.21-4.17(m,2H),4.15-4.08(m,1H),3.94-3.80(m,2H),3.78-3.58(m,2H),3.57-
3.48(m,3H),2.98-2.90(m,2H),2.88-2.84(m,2H),2.58-2.45(m,1H),2.25-2.13(m,2H),
2.09-1.93(m,2H),1.88-1.74(m,2H),1.70-1.48(m,4H),1.46-1.35(m,2H),1.35-1.22(m,
10H)；MALDI-TOF MS:[M+Na+]962.2350,[M+K+]978.2200
By intermediate product VI (855.0mg, 1.2120mmol), triethylamine (1.27mL, 12.12mmol) is added to 150mL
Flask in, use CH2Cl2(15mL) dissolving is stayed overnight, and solution turned cloudy adds triethylamine (2mL) to reacting complete, stops reaction,
Directly it is evaporated, uses CH2Cl2Dissolving, wet method loading, column chromatography first uses 200ml CH2Cl2Elution, then use 2%MeOH/CH2Cl2Arrive
4%MeOH/CH2Cl2Elution, finally uses 5%NH3·H2O/10%MeOH/CH2Cl2To 7.5%NH3·H2O/15%MeOH/
CH2Cl2Elution.Collect product and be evaporated to obtain product (for convenience of stating, being abbreviated as intermediate product VII) 885.1mg, yield 98.7%.
Proton nmr spectra such as Fig. 1：1H-NMR(500MHz,DMSO-d6)δ8.35-8.20(m,1H),8.15-7.78(m,3H),7.55-
7.35(m,4H),7.30-7.15(m,1H),6.85-6.42(m,2H),4.43-4.34(m,1H),4.33-4.21(m,1H),
4.20-4.12(m,1H),3.85-3.75(m,2H),3.74-3.64(m,2H),2.98-2.85(m,2H),2.64-2.53(m,
1H),2.28-2.18(m,2H),2.15-2.06(m,2H),1.93-1.84(m,2H),1.77-1.52(m,4H),1.50-1.41
(m,3H),1.40-1.35(m,12H)；MALDI-TOF MS:[M+H+] 718.3108, [M+Na+]740.2903
By intermediate product VII (360.0mg, 0.5020mmol), 4AMR-SCM-40K (2.510g, 0.06276mmol, north
Jing Jiankai Science and Technology Ltd.s) it is added in 100mL flasks, use CH2Cl2(5ml) dissolves, and is slowly stirred two weeks under room temperature condition,
It is transferred in 200mL flasks and is reacted again 7 days with DMF.Concentrated by rotary evaporation, then adds absolute ether sedimentation, filters to obtain crude product；So
Crude product dichloromethane and methanol are dissolved afterwards, ether sedimentation is added, filtration drying obtains VIII intermediate product 2.0578g, produces
Rate 78.8%.Proton nmr spectra such as Fig. 2：1H-NMR(500MHz,DMSO-d6)δ10.20-9.90(m,4H),8.40-8.20
(m,4H),8.10-7.91(m,8H),7.90-7.78(m,4H),7.65-7.58(m,4H),7.55-7.40(m,12H),7.31-
7.20(m,4H),6.82-6.51(m,4H),4.48-4.37(m,4H),4.34-4.27(m,4H),4.25-4.15(m,4H),
4.05-3.91(m,12H),3.85-3.60(m,24H),3.70-3.35(m,4199H),3.15-3.05(m,8H),3.03-
2.85(m,4H),2.35-2.20(m,8H),2.18-2.05(m,8H),1.96-1.84(m,8H),1.82-1.58(m,12H),
1.54-1.45(m,8H),1.44-1.31(m,44H)。
Intermediate product VIII (2.4g, 57.69mmol) is added in 30mL round-bottomed flasks, and uses CH2Cl2(20mL) dissolves,
Then add TFA (1mL) room temperature to be slowly stirred, stop reaction after two weeks, be directly evaporated, dissolved with dichloromethane (2mL), then
Add absolute ether precipitation.Filtering, filter cake is dried in vacuo to obtain intermediate product IX 1.6697g, yield 62.2%.Hydrogen nuclear magnetic resonance
Spectrum such as Fig. 3：1H-NMR(500MHz,CDCl3)δ9.98-9.82(m,4H),8.25-8.10(m,4H),7.78-7.55(m,4H),
7.48-7.20(m,8H),7.15-6.50(m,20H),6.20-6.10(m,4H),6.30-6.10,4.59-4.45(m,4H),
4.45-4.23(m,4H),4.23-4.02(m,12H),3.98-3.90(m,12H),3.85-3.40(m,4353H),2.54-
2.32(m,24H),2.28-2.06(m,16H),2.05-1.93(m,8H),1.86-1.70(m,8H),1.68-1.36(m,
32H)。
(1) by intermediate product IX (1.6697g, 0.0384mmol), succinyl amine alcohol (132.7mg, 1.1528mmol),
DMAP (4.7mg, 0.0384mmol) is added in 100ml round-bottomed flasks, uses CH2Cl2(400ml) dissolves, then by hybrid reaction
Liquid stirs 20min under the conditions of being placed in -5 DEG C, and DCC (237.9mg, 1.1528mmol) is added portionwise, and addition is moved to after finishing 1 hour
Room temperature, is slowly stirred reaction two weeks.Stop reaction, filtrate concentration is evaporated to obtain intermediate product X (1.6908g) by filtering.
(2) by intermediate product X (1.6908g, 0.0384mmol) and GEMCITABINE HYDROCHLORIDE (368.4mg,
1.2288mmol) it is dissolved in pyridine (10mL) in 100mL flasks, room temperature is slowly stirred two weeks, stops reaction, be directly evaporated,
Then dissolved, then be evaporated repeatedly 3 times with dichloromethane/ethyl acetate.Finally use CH2Cl2Dissolving, wet method loading uses MeOH/
CH2Cl2Elute successively, collect product point, the vacuum drying that is concentrated under reduced pressure obtains crude product, then dissolved with 2mL dichloromethane, added
Absolute ether precipitates it, filtering, is dried in vacuo to obtain finished product XI 1.2953g, yield 73.7%.For convenience of stating, by this end
Product XI is named as compound PEG (A+G).Proton nmr spectra such as Fig. 4：1H-NMR(500MHz,DMSO-d6)δ10.01-9.9
(m,4H),8.50-8.18(m,8H),8.16-7.95(m,8H),7.94-7.84(m,4H),7.82-7.70(m,8H),7.68-
7.60(m,4H),7.59-7.40(m,16H),7.38-7.20(m,4H),6.81-6.54(m,4H),6.38-6.21(m,4H),
6.00-5.30(m,4H),4.97-4.68(m,4H),4.65-4.35(m,8H),3.90-3.88(m,24H),3.85-3.42(m,
4263H),3.15-3.01(m,8H),2.87-2.72(m,16H),2.35-2.07(m,16H),2.05-1.83(m,16H),
1.80-1.58(m,8H),1.57-1.32(m,24H)；MALDI-TOF MS are in 37500-47500 scopes, top
42254.5313.Compound PEG (A+G) theoretical molecular：44497, wherein AZDD7762 theory drugloading rate are 3.257%,
And the theoretical drugloading rate of gemcitabine is 2.366%.
Embodiment 2
1. experiment reagent and experimental animal
The compound PEG (A+G) that embodiment 1 is obtained, compound are investigated on the Xenografts in nude mice model that p53 is mutated
A and antitumor drug effect associated with G.
Main agents
1640 culture medium：Hyclone, article No. SH30809.01B；
FBS：GIBCO, article No. 10099-141；
100* penicillin streptomycins are dual anti-：Invitrogen, article No. 10378-016；
PEG(A+G)：Light yellow translucent viscous solid, carries small molecule A medicines 3.257%, carries small molecule G medicines 2.366%,
Sample size about 1.3g；
A：AZD7762, molecular weight 362.4, white crystal；
G：Gemcitabine, molecular weight 263.2, GEMCITABINE HYDROCHLORIDE (G hydrochlorides) 299.65, white crystal；
The compound method of polyethylene glycol conjugation (gemcitabine and AZD7762) (being abbreviated as PEG (A+G) for convenience of statement)：
Appropriate test medicine is weighed, is dissolved, disperseed using sterile saline, laggard action thing administration is well mixed, now with existing
With.
A compound methods：Appropriate test medicine is weighed, dissolved using 12% HP- β-CD, disperse (suspension), mixed
Close uniform laggard action thing administration.
G compound methods：Appropriate test medicine is weighed, with physiological saline solution.
Experimental animal
Strain：Balb/c nude mices, SPF grades；Source：Shanghai Ling Chang bio tech ltd；Week old：5-6 weeks；Sex：
Male.
2. experimental method
PEG (A+G) tolerance studies
Under selected prescription, when dosage is 1540mg/kg, the tolerance situation to medicine is studied；Totally 4 it is naked
Mouse, i.p. is administered once, and is observed 4-5 days；
Colo-205 Xenografts in nude mice model is set up
A) recover and expand Colo-205 cells；
B) it is to be amplified to arrive enough cells, collect cell, be made into 1640 culture mediums without serum concentration for 2 ×
107Cells/ml cell suspension；
C) subcutaneous vaccination on the right side of nude mice, only, i.e., every nude inoculation cell number is 2 × 10 to 0.1ml/6It is individual；Inoculation 27.
Observation：Animal state is observed 2 times a week, goes out knurl situation, treats tumour growth to average external volume 100-200mm3Left and right
When packet administration.
Packet and administration：18 close animals of gross tumor volume are selected from 27 animals, are divided at random according to gross tumor volume
For 3 groups, respectively saline control group, PEG (A+G) group, small-molecule drug gemcitabine and the (letter of Chk1 inhibitor combinations group
It is written as A+G combinations), every group of 6 animals.The packet same day is designated as Day1, and the same day starts administration.Packet and former dosage regimen are shown in Table 1：
The animal packet of table 1. and administration
* G dosage is counted in a free form
Administration observation Day2, A+G groups 1#, 2#, 3#, 6# animal dead, 4 animals of selection add to this from standby animal
Group so that the tumor average volume of the group is suitable with saline control group；And former dosage regimen is revised, it is specifically shown in
Table 2：
Table 2:Animal packet is adjusted with administration
* G dosage is counted in a free form
Observation and detection：3-4 gross tumor volume and the weight of animals, measurement observation 14 days are measured weekly.
A+G combinations (G elder generations i.p. is administered, and A is administered in i.p. after 4h), according to former dosage regimen, (50+36.3) mg/kg gives
There is 4/6 animal dead in medicine next day；Therefore dosage is adjusted to (25+18.1) mg/kg, and according to the changes of weight of animal
Administration A, G dosing interval is adjusted to 48h (G elder generations i.p. is administered, and A is administered in i.p. after 48h).And PEG (A+G) is with 1540mg/
After kg is administered once, dosage substantially, therefore is adjusted to 770mg/kg by Body weight loss, and by dosing interval be adjusted to every 4 days to
Medicine 1 time.
3. Testing index and calculating, statistical analysis technique
Gross tumor volume (tumor volume, TV)
TV=1/2 × a × b2, wherein a, b represent length and width respectively.
Relative tumour volume (relative tumor volume, RTV)
RTV=TVt/TV1Wherein TV1(d1) gross tumor volume during for sub-cage administration, tumour when TVt is measures each time
Volume.
Relative tumor proliferation rate T/C (%)
TRTV：Treatment group RTV；CRTV：Blank control group RTV.
Statistical analysis：During experimental endpoints, the Levene that homogeneity of variance is carried out to each group relative tumour volume (RTV) (1) is examined
Test, if homogeneity of variance analyzes nonsignificance, i.e. P>0.05, using one-way analysis of variance, when variance analysis has notable meaning
Justice, i.e. P≤0.05, being examined from Dunnett will be compared between control group and other treatment group groups.(2) if Levene is examined
P≤0.05 tested, i.e. heterogeneity of variance, Logarithm conversion is carried out to the initial data without negative value, and the data after conversion carry out variance
The Levene of homogeneous is examined, if homogeneity of variance analyzes nonsignificance, i.e. P>0.05, using one-way analysis of variance, the side of working as
Difference analyses significance, i.e. P≤0.05, then is examined and will be compared between control group and other treatment group groups from Dunnett.
4. experimental result
PEG (A+G) tolerance studies
Under selected prescription, when dosage is 1540mg/kg, Body weight loss in 2 days after animal intraperitoneal injection,
Then have recovery trend, by such as table 3 of the observation of 1 week, medicine on the general state of animal without influence, including：Hair indifference；
Changes of weight no significant difference；Without nausea,vomiting,diarrhea, appetite, indigestion, dry skin, fash, red swelling of the skin, scratch
Itch, it is pain, backache, expiratory dyspnea, tired and situations such as have a sleepless night.Intracorporeal organ naked eyes viewing indifference.Mice sleep and
Enliven situation indifference；Mouse anus and defecation situation indifference, do not there is diarrhoea situation.
Influences (body weight and general state) of the PEG of table 3 (A+G) to animal
*-represent same day non-administration
Nude mice body weight and gross tumor volume result such as table 4：
The PEG of table 4 (A+G), A+G combination on the influence of Colo-205 tumor bearing nude mices body weight (data are represented using Mean ± SEM,
N=6)
a：Dosage regimen is specifically shown in Table 2；
b：The 3rd day, 2# animal deads is administered
c：It is administered the 2nd day, 1#, 2#, 3#, 6# animal deads, 4 animals is chosen from standby animal and add to the group；
RSD is respectively less than 20% in the group of each group gross tumor volume during this experiment packet, saline control during experimental endpoints
Group gross tumor volume reaches 1333.6mm3, gross tumor volume increases about 6.4 times when being relatively grouped, and meets transplanted tumor in nude mice drug efficacy study
General requirement.
The PEG of table 5 (A+G), A+G combination to Colo-205 transplanted tumor in nude mice grow inhibitory action (data using Mean ±
SEM represents, n=6)
a：Dosage regimen is specifically shown in Table 2；
b：Vs saline control groups, * p<0.05, * * p<0.01
Dosage regimen after being adjusted more than, after administration is observed 14 days, PEG (A+G) administration group shows significant anti-
Function of tumor, experimental endpoints Relative tumor proliferation rate T/C (%) is 59.9%；And the experimental endpoints of A+G combination administration groups are relative
Tumor proliferation rate T/C (%) is 72.0%.Fig. 5 is the tumour growth of PEG (A+G) group, A+G combinations group and saline control group
Curve.
The result of above-described embodiment 2 is shown：Saline control group the weight of animals is on a declining curve (with institute's lotus tumour phase
Close), and the decline of administration group the weight of animals is relatively more, is administered on the general state of animal without influence.To sum up, in Human colorectal carcinoma
On cell line Colo-205 Nude Mouse Models, under the dosage regimen of the present invention, PEG (A+G) is to Colo-205 nude mouses
Transplantable tumor, which has, significantly inhibits effect.
Claims (3)
1. a kind of high-molecular compound with active anticancer, its structural formula is as shown in XI：
2. the preparation method of high-molecular compound as claimed in claim 1, it is characterised in that：Comprise the following steps,
Under HBTU, HOBT, DIEA and solvent DMF, 0 DEG C to room temperature condition, Boc-Gly and Chk1 inhibitor AZD7762 is entered
Row amidatioon is connected, and is gone through TFA after Boc protections, the glutamic acid -5- tert-butyl esters of Gly-AZD7762 and the Fmoc protection of gained
In PyAOP and 2,4,6- trimethylpyridines, amidatioon connection is carried out under the conditions of 0 DEG C, then go Fmoc to protect with triethylamine and DMF
Shield, then the 6-aminocaprolc acid protected with Fmoc is in PyAOP and 2,4,6- trimethylpyridines, carries out amidatioon connection under the conditions of 0 DEG C,
Then Fmoc is gone to protect with triethylamine and DMF conditions again, gained micromolecular compound intermediate is high with quadrifurcate polyethylene glycol
Molecular vehicle 4ARM-SCM-40K carries out substituted amide connection, and gained high-molecular compound carries out the tert-butyl ester through TFA again and goes to protect
Shield, the hydroxy-acid group of gained aromatic polymer intermediates is again through n-hydroxysuccinimide, DCC, DMAP and CH2Cl2Condition is lived
Change, be finally coupled under the conditions of pyridine with gemcitabine, obtain final products XI.
3. application of the high-molecular compound as claimed in claim 1 in anticarcinogen is prepared.
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| CN107670048B (en) * | 2017-08-30 | 2019-04-26 | 南京明臻医药科技有限公司 | With the active intermediate drug of Synergistic anti-cancer and polyethylene glycol conjugation Synergistic anti-cancer drug, and its preparation method and application |
| CN111514309B (en) * | 2019-02-03 | 2021-09-17 | 重庆阿普格雷生物科技有限公司 | Polyethylene glycol coupled drug, preparation method and application thereof |
| CN112851928B (en) | 2019-11-28 | 2023-04-25 | 重庆阿普格雷生物科技有限公司 | Polyethylene glycol coupling drug, preparation method and application thereof |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103623394A (en) * | 2013-12-11 | 2014-03-12 | 南京安吉生物科技有限公司 | Application of PEG-HM-3 and platinum, taxol or Emtriva medicines to preparation of solid tumor medicines |
| CN104987504A (en) * | 2015-04-23 | 2015-10-21 | 南京明臻医药科技有限公司 | Pegylated lapatinib, injection and preparation method thereof |
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| CN103623394A (en) * | 2013-12-11 | 2014-03-12 | 南京安吉生物科技有限公司 | Application of PEG-HM-3 and platinum, taxol or Emtriva medicines to preparation of solid tumor medicines |
| CN104987504A (en) * | 2015-04-23 | 2015-10-21 | 南京明臻医药科技有限公司 | Pegylated lapatinib, injection and preparation method thereof |
Non-Patent Citations (4)
| Title |
|---|
| Edward Sausville et al.Phase I dose‑ * |
| escalation study of AZD7762, a checkpoint kinase inhibitor, in combination with gemcitabine in US patients with advanced solid tumors.《Cancer Chemother Pharmacol》.2014,第73卷第539-549页. * |
| Phase I, dose-escalation study of AZD7762 alone;Takashi Seto et al;《Cancer Chemother Pharmacol》;20131231;第72卷;第619–627页 * |
| Sensitization of Pancreatic;Venkatasubbaiah A. Venkatesha et al;《Neoplasia》;20121231;第14卷;第519–525页 * |
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