CN103570689A - Para-nitro aromatic methyl crizotinib hypoxia-activated prodrug for anticancer drugs - Google Patents

Para-nitro aromatic methyl crizotinib hypoxia-activated prodrug for anticancer drugs Download PDF

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CN103570689A
CN103570689A CN201310608734.3A CN201310608734A CN103570689A CN 103570689 A CN103570689 A CN 103570689A CN 201310608734 A CN201310608734 A CN 201310608734A CN 103570689 A CN103570689 A CN 103570689A
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fluorophenyl
piperidin
pyrazol
dichloro
pyridin
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CN103570689B (en
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何银杰
李万湖
齐放
张坤
孙学英
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JINAN TRIO PHARMACEUTICAL TECHNOLOGY Co Ltd
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Abstract

The invention relates to a para-nitro aromatic methyl crizotinib hypoxia-activated prodrug for anticancer drugs, and the chemical name is 5-(1-(1-(4-nitro aromatic methyl) piperidine-4-group)-1H-pyrazol-4-group)-3-((R)-1-(2,6-dichloro-3-fluorophenyl) ethyoxyl) piperidine-2-amine. A para-nitro aromatic methylation modifier is utilized to react with crizotinib for para-nitro aromatic methylation in an alkaline condition so as to prepare the crizotinib hypoxia-activated prodrug, para-nitro aromatic methyl crizotinib. The para-nitro aromatic methyl crizotinib hypoxia-activated prodrug is used as a main effective ingredient for preparing the anticancer drugs, and particularly for preparing the anticancer drugs for curing hepatocellular carcinoma.

Description

What be used for antitumor drug activates prodrug to nitro arylmethyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine anoxic
Technical field:
(7-ethyl-10-is to nitro benzyloxy (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine to nitro arylmethyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine anoxic activation prodrug to the present invention relates to new type antineoplastic medicine, abbreviation is to nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine, HAP-Crizotinib) preparation method, and their therapeutic actions to human body cancer especially liver cancer.
The advantage of nitro arylmethyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine is to it can activate under anoxic microenvironment specifically in tumor tissues, is converted into cytotoxic medicine.
Compare with (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine, nitro arylmethyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine is had to higher tumor-targeting and lower toxic side effect.
Background technology:
Crizotinib serving as antitumor molecular targeting medicament, has another name called Crizotinib, and chemical name is:
3-[(1R)-1-(the chloro-3-fluorophenyl of 2,6-bis-) oxyethyl group]-5-[1-(4-piperidines)-1H-pyrazoles-4-yl]-2-pyridine amine, molecular formula is:
Figure BDA0000422023210000011
(R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine is kinase inhibitor more than, can suppress Nucleophosmin-anaplastic lymphoma kinase (ALK), hepatocyte growth factor receptor (c-MET, HGFR) and the activity of tyrosine kinase receptor (RON), and then the propagation of these kinase whose tumour cells is expressed in blocking-up, induce its apoptosis, reach object (the Ou SH.Crizotinib:a novel and first-in-class multitargeted tyrosine kinase inhibitor for the treatment of anaplastic lymphoma kinase rearranged non-small cell lung cancer and beyond.Drug Des Devel Ther.2011 that suppresses tumor growth, 5:471-85.).
The above-mentioned kinases of kinds of tumor cells positive expression is found in research.
For example, in liver cancer patient; compare with surrounding normal liver organization; the liver cancer tissue high expression level c-MET of 30-100%; it will be a treatment target spot likely (Goyal L, et al.Targeting the HGF/c-MET Pathway in hepatocellular carcinoma.Clinical Cancer Research2013 that prompting suppresses c-Met; 19:2310-8.).
The activity that suppresses c-MET has significantly suppressed growth (You H, the et al.c-Met represents a potential therapeutic target for personalized treatment in hepatocellular carcinoma.Hepatology2011 of liver cancer tumour; 54:879-9).
Stomach cancer cell is high expression level c-MET also; therefore; (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine reaches by suppressing the activity of c-MET object (Okamoto W, et al.Antitumor action of the MET tyrosine kinase inhibitor crizotinib (PF-02341066) the in gastric cancer positive for MET amplification.Mol Cancer Ther2012 that suppresses Gastric Carcinoma Growth; 11:1557-64.).
The most significant example is nonsmall-cell lung cancer (the non-small-cell lung cancer that (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine has significantly suppressed the ALK positive, NSCLC) growth of cell, and in clinical trial, it has significantly improved the advanced NSCLC patient's of ALK high expression level and gene rearrangement survival rate (Helen Y Z, et al.Cancer Res., 2007,67 (9), 4408-4417).
On August 26th, 2011, (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine was used for the treatment of advanced Non-small cell lung by FDA (Food and Drug Adminstration) (FDA) approval, was current unique a kind of medicine for the treatment of such disease.
Yet as the first ALK inhibitor for oral use in the whole world, the common adverse reactions (>=25%) of (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine is visual disorder, feel sick, diarrhoea, vomiting, oedema and constipation.
Liver power checking once found that patient ALT and the total bilirubin of taking (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine raise simultaneously, if these indexs continue to raise, need to temporarily stop, and lowered dosage, or forever stopped taking.
The appearance of resistance situation simultaneously has seriously limited its clinical application; In addition, (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine solution is unstable at normal temperatures, water-soluble poor, and in aqueous medium, within the scope of pH1.6～pH8.2, (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine solubleness is reduced to and is less than 0.1mg/mL from being greater than 10mg/mL.
Therefore, in order to reduce toxic side effect and the serious adverse reaction of (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine, increase tumor-targeting, improve its stability and water-soluble, increase its bioavailability, we have carried out structural modification to (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine and have had the (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine derivative compared with high biological activity and stability to obtaining.
Summary of the invention:
This invention for be the anoxic microenvironment comprising in nearly all mankind's noumenal tumour and a kind of novel (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine anoxic researched and developed activates prodrug.
The object of the invention is synthetic a kind of new type antineoplastic medicine (to nitro arylmethyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine) based on antineoplastic target small-molecule drug (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine.
This new synthetic anoxic activates prodrug and compares with (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine and has better water-soluble and biologically stable, and it is not normally activated oxygen level in the situation that in body member tissue, low to the toxic side effect of liver and enteron aisle; And under the inner distinctive anoxic microenvironment of noumenal tumour (comprising the modal liver cancer of China, cancer of the stomach, lung cancer, colorectal cancer, mammary cancer, intestinal cancer, prostate cancer etc.), be selectively activated, be converted into the (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine with killing functions of immunocytes, thereby reach the object of optionally killing cancer cells.
Therefore, compare with the (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine of current clinical application, have advantages of that tumor-selective is strong, toxicity is low, bioavailability is high.
The chemical name to nitro arylmethyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine anoxic activation prodrug for liver cancer treatment provided by the present invention is: 5-(1-(1-(4-nitro arylmethyl) piperidin-4-yl)-1H-pyrazoles-4-yl)-3-((R)-1-(2, the chloro-3-fluorophenyl of 6-bis-) oxyethyl group) pyridine-2-amine; Referred to as to nitro arylmethyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine or 4-nitro arylmethyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine.
Its structural formula is as follows:
Figure BDA0000422023210000041
Y=H,F,Cl,Br,R,OR,SR,CN,CONH 2,COR,COOR,CF 3,SO 2R,NO 2or
Figure BDA0000422023210000042
In above formula, Y can replace on 2 or 3 positions of phenyl ring, can be also two replacement; Y can be as fluorine, chlorine, bromine, methyl, methoxyl group, first sulfydryl etc.; Y can be also cyano group, amide group, formyl radical, methoxycarbonyl, trifluoromethyl, methylsulfonyl and nitro etc.
Y can also be dimethylin, piperidyl, N '-methylpiperazine base, the pyridyl (side chain position can be 2,3,4) that 0-6 methylene radical connects.
R is C1-C6 fat group or fragrant group.
By changing the substituting group on phenyl ring, change the redox potential of nitro arylmethyl, since regulate nitro arylmethyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine emitted to the difficulty or ease of (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine under anaerobic environment, reach for the best anoxic of different carcinoma cell screening optionally to nitro arylmethyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine, simultaneously by increasing substituent polarity, improve that it is water-soluble.
Prodrug---the preparation method to nitro arylmethyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine is as follows in (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine anoxic activation of the present invention:
Utilization, to nitro arylmethyl modifier, under alkaline condition, is reacted with (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine, to it to nitro arylmethyl.
Its chemical equation is as follows:
Figure BDA0000422023210000051
In said structure formula, X can be fluorine, chlorine, bromine or iodine; Y can replace on 2 or 3 positions of phenyl ring, can be also two replacement; Y can be as fluorine, chlorine, bromine, methyl, methoxyl group, first sulfydryl etc.; Y can be also cyano group, amide group, formyl radical, methoxycarbonyl, trifluoromethyl, methylsulfonyl and nitro etc.
Y can also be dimethylin, piperidyl, N '-methylpiperazine base, the pyridyl (side chain position can be 2,3,4) that 0-6 methylene radical connects.
R is C1-C6 fat group or fragrant group.
Described is to utilize nitro arylmethyl modifier under alkaline condition to nitro arylmethyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine, reacts it is made nitro arylmethyl with (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine.
The above-mentioned preparation process to nitro arylmethyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine is as follows:
To be dissolved in solvent nitro arylmethyl modifier and (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine, add alkali at 25 ℃, and add latter 25 ℃ and stir 3 hours; Add water extraction, separate organic phase, water layer dichloromethane extraction, merge organic phase anhydrous sodium sulfate drying, removal of solvent under reduced pressure, obtains methylene dichloride and isopropyl ether recrystallization for thick product, it is yellow solid that filtration obtains sterling, is nitro arylmethyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine.
In above-mentioned preparation method, what use can be to nitro arylmethyl muriate, bromide, iodide or methylsulfonic acid virtue methyl esters to nitro arylmethyl modifier; To the mol ratio of nitro arylmethyl modifier and (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine, can be 1.0-2.0; The alkali using can be salt of wormwood, diisopropylethylamine, sodium carbonate and triethylamine; Solvent can be selected ether, methylene dichloride, tetrahydrofuran (THF) and acetonitrile etc.
Now take unsubstituted (Y=H) simply to nitro arylmethyl modifier---to the modification reaction of nitrobenzyl modifier and (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine, be example, inquire into its concrete preparation process and processing condition, owing to having impurity in reaction process, generate, so the kind of modifier, solvent, alkali and consumption proportion are carried out preferably:
Its reaction formula is as follows:
1) to nitrobenzyl modifier kind preferably:
First, by the exploration of tentative experiment, tentative alkali is triethylamine, 2 moles of consumptions; Methylene chloride, consumption is 15 mls/g of Crizotinibs; To nitrobenzyl modifier and Crizotinib mol ratio, be 1.5:1; Temperature of reaction is 25 ℃, and the time is 3 hours; Test determination difference is the impact on product yield on nitrobenzyl modifier.
Result is as table 1:
The impact of the different types of modifier of table 1. on product yield
Figure BDA0000422023210000062
Figure BDA0000422023210000071
As can be seen from Table 1, the activity of p-nitrobenzyl bromide is higher, and reaction yield is also higher, although the activity to nitro iodate benzyl is higher than p-nitrobenzyl bromide, but because it is too active, thus some can be generated by the alkylating impurity in other positions, and cause yield to reduce.
2) p-nitrobenzyl bromide consumption is preferred:
Confirm that p-nitrobenzyl bromide is preferred modifier, above-mentioned 1) in the constant situation of other conditions, the preferred consumption of p-nitrobenzyl bromide.
Result is as table 2:
The impact of table 2. nitro benzyl bromide consumption on product yield
Figure BDA0000422023210000072
As can be seen from Table 2, the consumption of p-nitrobenzyl bromide is larger, and yield is higher, and impurity also can be more.
When surpassing 1.5 moles, yield increases not obvious, considers cost factor, and the consumption of p-nitrobenzyl bromide is preferably 2 moles.
3) kind of alkali is preferred:
Determine that p-nitrobenzyl bromide is 1.5 moles of modifier consumptions; In the constant situation of other conditions, use the alkali of different varieties in reaction, the product yield of acquisition is in Table 3.
The impact of the different types of alkali of table 3. on product yield
Figure BDA0000422023210000081
As can be seen from Table 3, when using triethylamine to do alkali, its alkalescence is better than diisopropylethylamine, can make reaction yield be significantly improved.
When using mineral alkali sodium carbonate and salt of wormwood, because its solvability is bad, so yield is not used organic bases yield high.
4) triethylamine consumption is preferred:
In the situation that other conditions are constant, the consumption of preferred triethylamine.
The results are shown in Table 4.
The impact of table 4. triethylamine consumption on product yield
As can be seen from Table 4, triethylamine consumption should be preferably 2 moles.
5) solvent kind is preferred:
Determine that p-nitrobenzyl bromide is preferred modifier, its Determination of quantity is 2 moles, in the situation that other conditions are constant, and preferred solvent kind.
Result is as table 5:
The impact of table 5. solvent on product yield
Figure BDA0000422023210000091
As can be seen from Table 5, this reaction needed is carried out in middle polarity non-protonic solvent, can reduce the selectivity of reaction with the large solvent of polarity, and the solvent that polarity is less is unfavorable for again the carrying out of reaction.
With the very large acetonitrile of polarity or polarity ether less than normal, all can reduce the yield of reaction, only select methylene dichloride yield the highest.
6) solvent load is preferred
Determine that methylene dichloride is solvent, in the situation that other conditions are constant, preferred solvent consumption.
The results are shown in Table 6:
The impact of table 6. solvent load on product yield
Figure BDA0000422023210000092
As can be seen from Table 6, solvent load is on the low side, reacts influenced, and yield is on the low side, because concentration is when larger, the selectivity of reaction is poor, and impurity generates more.
Consumption yield when 15ml/g (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine is higher, and when consumption is increased to 20ml/g (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine, yield has downtrending on the contrary, when to be that solvent load is too high cause purifying products to reclaim, causes damage.
7) p-nitrobenzyl bromide and triethylamine usage ratio is preferred
Determine that methylene dichloride is solvent, in the situation that other conditions are constant, preferred p-nitrobenzyl bromide and triethylamine usage ratio.
The results are shown in Table 7:
Table 7. p-nitrobenzyl bromide and the impact of triethylamine usage ratio on product yield
Figure BDA0000422023210000101
As can be seen from Table 7, p-nitrobenzyl bromide and triethylamine consumption are more approaching better, but reaction needed just can carry out under alkaline environment, because alkalescence is too strong, the content of impurity can increase.
In order to keep the alkalescence of system, the consumption of triethylamine must be higher than the consumption of p-nitrobenzyl bromide, and the two ratio is preferably 0.75:1.
By above-mentioned optimization test, contrast, confirm that preparation method's of the present invention preferred processing condition is as follows:
Nitrobenzyl modifier is selected to p-nitrobenzyl bromide, and consumption is 1.0-1.8 mole (being optimized for 1.5 moles); The variety adoption triethylamine of alkali, consumption is 1.0-2.5 mole (being optimized for 2 moles); P-nitrobenzyl bromide and triethylamine usage ratio are preferably 0.75:1; Solvent is selected methylene dichloride, and consumption is (5-20ml/g (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine) (preferably 15ml/g (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine); Temperature of reaction is optimized for 25 ℃, 3 hours time; What under above-mentioned optimum condition, reaction made is more satisfactory to nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine yield.
Of the present invention nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine be can be used as to main effective constituent for the preparation of anti-tumor agents; Especially for the anti-tumor agents of preparing Hepatoma therapy treatment.
Beneficial effect to nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine of the present invention is as follows:
This invention nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine is compared with the (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine that is applied at present cancer therapy, there is significantly lower toxic side effect, be mainly manifested in the degree that loses weight after, diarrhoea mild degree low to the infringement of intestinal mucosa, treatment little, low to liver function impairment.
Give after two kinds of medicines, the change in concentration of the (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine in blood plasma is little, but significantly high than (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine treatment group to (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine concentration in nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine treatment group tumor tissues, and to (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine concentration in nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine treatment group liver and small intestine significantly lower than (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine treatment group.
Show that the present invention has less toxic side effect and stronger tumor-targeting to nitrobenzyl to nitrobenzyl.
Accompanying drawing explanation:
Fig. 1 is the nucleus magnetic hydrogen spectrum figure to nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine
Fig. 2. be rat liver cancer tumor growth correlation curve
Fig. 3. be the content balance of HAP-Crizotinib and Crizotinib in mouse implantation tumor, liver and small intestine and blood
Fig. 4. be Mouse Weight correlation curve figure
Fig. 5. be mouse jejunal mucous membrane damage comparative analysis
Fig. 6. be the comparative analysis of mouse liver function
Embodiment:
1. pairs of nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amines of embodiment and preparation method thereof
1) chemical name to nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine:
5-(1-(1-(4-nitrobenzyl) piperidin-4-yl)-1H-pyrazoles-4-yl)-3-((R)-1-(2, the chloro-3-fluorophenyl of 6-bis-) oxyethyl group) pyridine-2-amine.
Chemical structural formula to nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine:
Figure BDA0000422023210000121
2) the preferred preparation method to nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine:
3.25 grams of p-nitrobenzyl bromides and 4.5 grams of (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amines are dissolved in 67.5 milliliters of methylene dichloride, drip 2.02 grams of triethylamines at 25 ℃, add latter 25 ℃ and stir 3 hours.
75 ml waters, separate organic phase, dichloromethane extraction for water layer (50 milliliters * 3).
Merge organic phase anhydrous sodium sulfate drying, removal of solvent under reduced pressure, obtains methylene dichloride and isopropyl ether recrystallization for thick product, and filtering and obtaining sterling is 5.09 grams of yellow solids, yield 87%.
3) as follows to the nucleus magnetic hydrogen spectrum of nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine: (seeing accompanying drawing 1)
1HNMR(600MHz,DMSO-d 6)δppm8.22(dt,J=9.0,1.8Hz,2H),7.98(s,1H),7.75(d,J=1.5Hz,1H),7.62(d,J=8.4Hz,2H),7.57(dd,J=9.0,4.8Hz,1H),7.53(s,1H),7.45(t,J=9.0Hz,1H),6.90(d,J=1.8Hz,1H),6.08.(q,J=6.6Hz,1H),5.65(s,2H),4.11_4.16(m,1H),3.66(s,2H),2.88(d,J=11.4Hz,2H),2.19(td,J=10.8,3.0Hz,2H),1.94_2.01(m,4H),1.80(d,J=6.6Hz,3H).
Other preparation method's contrasts of 2. pairs of nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amines of embodiment:
4.73 grams are dissolved in 90 milliliters of tetrahydrofuran (THF)s nitro iodate benzyl and 4.5 grams of (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amines, drip 3.23 grams of diisopropylethylamine at 25 ℃, add latter 25 ℃ and stir 3 hours.
75 ml waters, separate organic phase, dichloromethane extraction for water layer (50 milliliters * 3).
Merge organic phase anhydrous sodium sulfate drying, removal of solvent under reduced pressure, obtains methylene dichloride and isopropyl ether recrystallization for thick product, and filtering and obtaining sterling is 3.16 grams of yellow solids, yield 54%.
The effect of 3. pairs of nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amines of embodiment and with the contrast of (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine
1) comparative analysis to the evaluation of nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine antitumour activity and standard drug (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine.
At female Balb/c nude mice in 6 week age left side flank hemostasis 5 * 10 6the human liver cancer cell MHCC97-H of logarithmic phase and Hep3B cell are in left side side of body portion.
Wherein, MHCC97-H cell high expression level c-MET; and the low expression of Hep3B cell c-MET(You H, et al.c-Met represents a potential therapeutic target for personalized treatment in hepatocellular carcinoma.Hepatology2011; 54:879-9).
When tumour grows to 100mm 3(day0), animal is divided into three groups at random, it is control group, (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine group and to nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine group, give respectively intubate stomach and raise 0.5% the Vltra tears aqueous solution, (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine (50mg/kg) and to nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine (50mg/kg), once a day, continues 3 weeks.
Tumor size calculates by measuring major diameter (L) and the minor axis (W) of tumour: LW 2/ 2.
Tumor size and body weight do not have three days and measure once.
From growth curve chart (Fig. 2 A and B), can find out, (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine and to nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine treatment group, the growth of the Hep3B liver cancer tumour of low expression c-MET is slower than control group (P value is all less than 0.05) (Fig. 2 A); (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine and to nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine treatment group, the growth of the MHCC97-H liver cancer tumour of high expression level c-MET is significantly slower than control group (P value is all less than 0.001) (Fig. 2 B).
Illustrate that (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine can suppress by suppressing the activity of c-MET the growth of liver cancer tumour.
Yet two kinds of tumor sizes, at (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine and almost identical to nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine treatment group, do not have significant difference, illustrate nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine similar to (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine in the curative effect of Hepatoma therapy.
2) (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine and to the tissue distribution comparative analysis in animal body of nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine.
Above-mentioned 1) accept (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine and the mouse of nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine treatment is put to death in treatment for 21 days, leaving and taking blood, liver, small intestine and tumor tissues.
Utilize high-efficient liquid phase chromatogram technology to measure the concentration of (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine in tumour, liver, small intestine and serum.
As shown in Figure 3, in (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine treatment group animal plasma, (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine concentration is 1198ng/mL, to nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine treatment group (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine concentration, be 875ng/mL, (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine treatment group, higher than to nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine treatment group, is 1.37 times (P<0.05) (Fig. 3 A) of the latter; (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine concentration in nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine treatment group tumor tissues is significantly higher than to (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine treatment group, 1.68 times (P<0.001) (Fig. 3 B) that the former is the latter; Yet the concentration of the (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine in (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine treatment group liver and small intestine is significantly higher than nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine treatment group (P<0.001) (Fig. 3 C); This results verification, has stronger tumor-selective to nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine; Make its in tumor tissues under anaerobic environment oxidized reduction discharge (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine.
3) (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine and the comparative analysis to Mouse Weight, diarrhoea, mucous membrane of small intestine infringement and liver function to nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine.
To above-mentioned 1) accept (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine and the body weight of the mouse of nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine treatment is carried out to record.
As shown in Fig. 4 A and B, compare with control group, (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine and nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine treatment group Mice Body weight average is obviously declined, and the body weight of (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine treatment group mouse is starkly lower than nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine treatment group (P value is all less than 0.05).
In order further to observe the diarrhea of mouse that (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine causes, the present invention tests as follows: 8 week age, female C57B/L mouse was divided into three groups (10 every group): control group, (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine group and to nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine group, carrying out respectively intubate stomach raises: 0.5% the Vltra tears aqueous solution, (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine (200mg/kg) and to nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine (200mg/kg), once a day, continue 1 week.
The 7th day, observe animal diarrhoea situation, and the severity of diarrhoea is carried out to sxemiquantitative counting by following standard.
As shown in table 8, control animals stool is normal, does not occur any unusual phenomenon; And (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine with nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine treatment group animal all be there is to the diarrhoea that degree is different; To the diarrhoea degree of nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine treatment group animal significantly lower than (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine treatment group (P<0.05).
The variation of the weight of animals and diarrhoea degree have been reacted the function that pipe intestinal digesting absorbs, and for the further damage to small intestine, do objective analysis, and the present invention has carried out following detection to the jejunum of above-mentioned mouse: the height of measuring jejunum villi under microscope; Measure the activity of γ-GGT in intestinal tissue.
As shown in Figure 5A, (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine and to the height of nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine treatment group mouse jejunal mucous membrane fine hair all lower than control group, and the highly significant of nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine treatment group mouse jejunal mucous membrane fine hair is greater than to (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine treatment group (P<0.05).
(R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine and organize the activity of γ-GGT all lower than control group to nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine treatment group mouse jejunum, and organize the activity of γ-GGT to be significantly greater than (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine treatment group (P<0.05) (Fig. 5 B) to nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine treatment group mouse jejunum.
Liver power checking finds that patient ALT and the total bilirubin of taking (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine raise, and therefore, we check the Serum ALT of above-mentioned mouse and total bilirubin.
As shown in Figure 6A, (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine and to nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine treatment group mice serum ALT level all higher than control group, and to nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine treatment group ALT level significantly lower than (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine treatment group (P<0.05).
(R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine and to nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine treatment group mice serum total bilirubin level all lower than control group, and to nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine treatment group total serum bilirubin significantly lower than (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine treatment group (P<0.05) (Fig. 6 B).
Above results suggest, compares with (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine, and nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine is obviously reduced the detrimental effect of gi tract and liver.
Table 8. diarrhea of mouse grade
Figure BDA0000422023210000151

Claims (10)

1. one kind is activated prodrug for antitumor drug to nitro arylmethyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine anoxic, its chemical name is: 5-(1-(1-(4-nitro arylmethyl) piperidin-4-yl)-1H-pyrazoles-4-yl)-3-((R)-1-(2, the chloro-3-fluorophenyl of 6-bis-) oxyethyl group) pyridine-2-amine;
Chemical structural formula is:
Figure FDA0000422023200000011
Y=H,F,Cl,Br,R,OR,SR,CN,CONH 2,COR,COOR,CF 3,SO 2R,NO 2or
Figure FDA0000422023200000012
R is C1-C6 fat group or fragrant group.
2. as claimed in claim 1 to nitro arylmethyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine anoxic activation prodrug, it is characterized in that utilizing nitro arylmethyl modifier, under alkaline condition, react with (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine (Crizotinib), make it to nitro arylmethyl, make (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine anoxic and activate prodrug---to nitro arylmethyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine (HAP-Crizotinib).
3. the preparation method who nitro arylmethyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine anoxic is activated to prodrug as claimed in claim 1, is characterized in that to nitro arylmethyl modifier and (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine to be dissolved in solvent, adds alkali at 25 ℃, adds latter 25 ℃ and stirs 3 hours; Add water extraction, separate organic phase, water layer dichloromethane extraction, merge organic phase anhydrous sodium sulfate drying, removal of solvent under reduced pressure, obtains methylene dichloride and isopropyl ether recrystallization for thick product, it is yellow solid that filtration obtains sterling, is nitro arylmethyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine.
4. the preparation method who nitro arylmethyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine anoxic is activated to prodrug as claimed in claim 1, is characterized in that used can be to nitro arylmethyl muriate, bromide, iodide or methylsulfonic acid virtue methyl esters to nitro arylmethyl modifier; To the mol ratio of nitro arylmethyl modifier and (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine, can be 1.0-1.8; The alkali using can be salt of wormwood, diisopropylethylamine, sodium carbonate and triethylamine; Solvent can be selected ether, methylene dichloride, tetrahydrofuran (THF) and acetonitrile.
5. as claimed in claim 1 or 2 nitro arylmethyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine anoxic is activated to prodrug, it is characterized in that utilizing p-nitrobenzyl bromide for modifier, react and make with (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine, its chemical name is:
5-(1-(1-(4-nitrobenzyl) piperidin-4-yl)-1H-pyrazoles-4-yl)-3-((R)-1-(2, the chloro-3-fluorophenyl of 6-bis-) oxyethyl group) pyridine-2-amine; Abbreviation is to nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine or 4-nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine;
Chemical structural formula is:
Figure FDA0000422023200000021
6. as claimed in claim 5 to nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine anoxic activation prodrug, it is characterized in that the nucleus magnetic hydrogen spectrum of nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine as follows:
1HNMR(600MHz,DMSO-d 6)δppm8.22(dt,J=9.0,1.8Hz,2H),7.98(s,1H),7.75(d,J=1.5Hz,1H),7.62(d,J=8.4Hz,2H),7.57(dd,J=9.0,4.8Hz,1H),7.53(s,1H),7.45(t,J=9.0Hz,1H),6.90(d,J=1.8Hz,1H),6.08.(q,J=6.6Hz,1H),5.65(s,2H),4.11_4.16(m,1H),3.66(s,2H),2.88(d,J=11.4Hz,2H),2.19(td,J=10.8,3.0Hz,2H),1.94_2.01(m,4H),1.80(d,J=6.6Hz,3H)。
7. the preparation method who nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine anoxic is activated to prodrug as claimed in claim 5, is characterized in that processing condition are as follows:
Nitrobenzyl modifier is selected to p-nitrobenzyl bromide, and consumption is 1.0-1.8 mole; The variety adoption triethylamine of alkali, consumption is 1.0-2.5 mole; P-nitrobenzyl bromide and triethylamine usage ratio are preferably 0.75:1; Solvent is selected methylene dichloride, and consumption is (5-20) ml/g (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine; Temperature of reaction is optimized for 25 ℃, 3 hours time.
8. as described in claim 5 or 7, nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine anoxic is activated to the preparation method of prodrug, it is characterized in that processing condition are as follows:
Nitrobenzyl modifier is selected to p-nitrobenzyl bromide, and consumption is 1.5 moles; The variety adoption triethylamine of alkali, consumption is 2 moles; P-nitrobenzyl bromide and triethylamine usage ratio are preferably 0.75:1; Solvent is selected methylene dichloride, and consumption is 15ml/g (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine; Temperature of reaction is optimized for 25 ℃, 3 hours time.
9. the preparation method who nitrobenzyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine anoxic is activated to prodrug as claimed in claim 5, is characterized in that concrete preparation method is as follows:
3.25 grams of p-nitrobenzyl bromides and 4.5 grams of (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amines are dissolved in 67.5 milliliters of methylene dichloride, at 25 ℃, drip 2.02 grams of triethylamines, add latter 25 ℃ and stir 3 hours, 75 ml waters, separate organic phase, dichloromethane extraction for water layer (50 milliliters * 3), merge organic phase anhydrous sodium sulfate drying, removal of solvent under reduced pressure, obtains methylene dichloride and isopropyl ether recrystallization for thick product, it is 5.09 grams of yellow solids that filtration obtains sterling, yield 87%.
10. the purposes to nitro arylmethyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine anoxic activation prodrug as described in claim 1 or 5, is characterized in that as main effective constituent for the preparation of anti-tumor agents, especially for the anti-tumor agents of preparation treatment hepatocellular carcinoma.
CN201310608734.3A 2013-11-25 2013-11-25 For antitumor drug to nitro arylmethyl (R)-3-(1-(2,6-dichloro-3-fluorophenyl)ethoxy)-5-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)pyridin-2-amine anoxic activation prodrug Active CN103570689B (en)

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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101321527A (en) * 2005-12-05 2008-12-10 辉瑞产品公司 Method of treating abnormal cell growth
CN102731519A (en) * 2012-06-26 2012-10-17 济南精合医药科技有限公司 P-nitro aryl methoxycamptothecine anoxic activated prodrug used for antitumor drug

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101321527A (en) * 2005-12-05 2008-12-10 辉瑞产品公司 Method of treating abnormal cell growth
CN102731519A (en) * 2012-06-26 2012-10-17 济南精合医药科技有限公司 P-nitro aryl methoxycamptothecine anoxic activated prodrug used for antitumor drug

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