CN102603761A

CN102603761A - Etoposide compound with hydroxamic acid structure and preparation method and usage thereof

Info

Publication number: CN102603761A
Application number: CN201210023070XA
Authority: CN
Inventors: 吕伟; 张翾; 罗宇; 陈奕; 李佳; 李静雅; 丁健
Original assignee: Shanghai Institute of Materia Medica of CAS; East China Normal University
Current assignee: Shanghai Institute of Materia Medica of CAS; East China Normal University
Priority date: 2012-02-02
Filing date: 2012-02-02
Publication date: 2012-07-25
Anticipated expiration: 2032-02-02
Also published as: CN102603761B

Abstract

The invention provides a novel etoposide derivative with a hydroxamic acid structure as showed in the general formula I, a pharmaceutically-acceptable salt of the etoposide derivative, a preparation method of the etoposide derivative and usage of the etoposide derivative. In the general formula I, X is -NH-, -O-, -S-, -NH-CH2-, -O-CH2- or -S-CH2-; Y is aromatic ring, substituted aromatic ring, aromatic hyterocyclic ring or substituted aromatic hyterocyclic ring; Z is -O-, -NHCO- or -CONH-; and B is C1-C8 alkyl, C2-C8 alkenyl, alkynyl or cycloalkyl. The compound provided by the invention has good antitumor activity and can be applied clinically by oral administration, intravenous injection or intramuscular injection.

Description

The Etoposide compound and preparation method and the purposes that contain the hydroxamic acid structure

Technical field

The present invention relates to one type of many target spots suppressor factor that acts on topoisomerase II and histon deacetylase (HDAC), and the method and the purposes that prepare these compounds.More particularly, relate to novel epipodophyllotoxin derivatives with following structure ( I) and their preparation method and these compounds in antitumor Application for Field.

Figure 201210023070X100002DEST_PATH_IMAGE002

Background technology

The DNA topoisomerase is one type of important antitumor target spot, podophyllotoxin ( 1) be the representational antineoplastic compound that acts on topoisomerase II; From mayapple, separate the earliest and obtain from kindred plants such as Sixangular Dysosma Rhizome and Root, Tibet Podophyllum emodi var chinense and unmrellaleaf, to obtain again afterwards; It has significant antitumor and antiviral activity, but greatly its application is restricted because of strong toxicity, spinoff.

By the verivate VP of podophyllotoxin structure of modification gained (etoposide, VP-16) ( 2) and phosphoric acid salt (etoposide phosphate, or claim etopophos) and teniposide (teniposide, VM-26) ( 3) become and be applied to clinical anticancer representative medicine.They all have good curative effect to multiple cancers such as small cell lung cancer, carcinoma of testis, acute leukemia and malignant lymphomas.This compounds can act on topoisomerase II, forms cleavability mixture stable between a kind of medicine-enzyme-DNA three, disturbs the DNA topoisomerase II, causes impaired DNA not repair, thus cell death inducing.

Figure 201210023070X100002DEST_PATH_IMAGE004

A new trend of antitumor drug research is the single medicine that development can act on a plurality of target spots simultaneously.Because blocked many approach of tumour cell existence, this small-molecule drug can overcome traditional medicine and be easy to generate chemical sproof shortcoming.For example, first-generation tyrosine kinase inhibitor imatinib action target spot is comparatively single, mainly suppresses Tyrosylprotein kinase Bcr-Abl.Patient is easy to generate resistance after taking the imatinib of for some time.S-generation tyrosine kinase inhibitor nilotinib and Dasatinib have the ability of abl and the dual kinase inhibition of src, can treat imatinib is produced drug-fast patient, thereby demonstrate than the better result of treatment of first-generation medicine.

Mammiferous albumen deacetylase (HDAC) is one type of hydroamidase that contains zine ion, can be divided into the first kind (HDAC-1, HDAC-2, HDAC-3 and HDAC-8), second type (HDAC-4, HDAC-5, HDAC-7, HDAC-9, HDAC-10) and the 4th type (HDAC-11).NSC 630176 (HDACi) can suppress white blood disease and solid tumor cell propagation, and the inducing cell differentiation.In addition, hdac inhibitor can make normal cell produce a G2 phase outpost of the tax office, makes normal cell to withdraw from the cell cycle with a kind of safe mode, and this is that hdac inhibitor is to one of very little reason of Normocellular toxic side effect.

The present invention combines hdac inhibitor and podophyllotoxin analogue, has developed the novel hybride molecule, has dual anti-tumor activity.

Summary of the invention

The objective of the invention is to seek the one type of novel segmental epipodophyllotoxin derivatives of hydroxamic acid and pharmacy acceptable salt thereof of containing, this compound possesses topoisomerase II and the effect of histon deacetylase (HDAC) double inhibition.

Another object of the present invention provides a kind of method for preparing said novel Etoposide chlorins compound and pharmacy acceptable salt thereof.

Another object of the present invention provides a kind of compsn that is used to treat cancer, and said composition comprises one or more said Etoposide chlorins compounds and the pharmacy acceptable salt and the pharmaceutically acceptable carrier of treating significant quantity.Said pharmacy acceptable salt includes but not limited to the additive salt of this compound and hydrochloric acid, sulfuric acid, phosphoric acid, methylsulfonic acid, succsinic acid, toxilic acid, fumaric acid, tartrate, Hydrocerol A etc.

A purpose more of the present invention provides a kind of said Etoposide chlorins compound and the application of pharmacy acceptable salt in the preparation antitumor drug thereof.The antitumor drug of target spot more than this type has wider antitumor spectrum than original medicine, overcomes the resistance of some tumor line to these two kinds of medicines.

The invention relates to the epipodophyllotoxin new derivatives, these compounds are to introduce the hydroxamic acid fragment in the C4 position of epipodophyllotoxin, and connect through rational fragment.They have good antineoplastic activity.

The segmental Etoposide chlorins compound of hydroxamic acid and the salt concrete structure pharmaceutically thereof of containing of the present invention is shown in general formula (I):

Wherein,

X is-NH-,-O-,-S-,-NH-CH ₂-,-O-CH ₂-or-S-CH ₂-;

Y is aromatic nucleus, substituted aromatic nucleus, fragrant heterocycle or substituted fragrant heterocycle; Be preferably phenyl ring or 1,2, the 3-triazole;

Z is-O-,-NHCO-or-CONH-; Be preferably-O-;

B is the alkyl of C1-C8, alkenyl, alkynyl or the naphthenic base of C2-C8, is preferably the alkyl of C3-C7.

The invention provides a kind of described preparation method who contains segmental Etoposide chlorins compound of hydroxamic acid and pharmaceutically-acceptable salts thereof, this method comprises the steps:

Wherein, n=3-7;

A) compound I I reacts with sulfur oxychloride in methyl alcohol, gets compound III; This step temperature of reaction is 0-40 ℃;

B) under the alkali action condition, compound III and nitrophenols reaction obtain compound IV; Alkali is potassium tert.-butoxide, sodium tert-butoxide, yellow soda ash, cesium carbonate, salt of wormwood etc., is preferably salt of wormwood, and reaction solvent is THF, N, and dinethylformamide, methyl-sulphoxide etc. are preferably N;

C) under the alkali action condition, the compound IV hydrolysis gets compound V; Alkali is sodium hydroxide, Pottasium Hydroxide, Lithium Hydroxide MonoHydrate, salt of wormwood etc., is preferably sodium hydroxide; Reaction solvent is alcohol-water, methanol-water, THF-water, is preferably alcohol-water;

D) under anhydrous condition, compound V and sulfur oxychloride reaction get compound VI; Reaction solvent is methylene dichloride, toluene; Temperature of reaction is 0-80 ℃, is preferably 40-60 ℃;

E) under alkali and catalyzer acting in conjunction, compound VI and N-boc-O-TBS azanol reaction get compound VI I; Alkali is triethylamine, pyridine, Diethyl Aniline etc., is preferably triethylamine; Catalyzer is a Dimethylamino pyridine; Temperature of reaction is-20-20 ℃, be preferably-20-0 ℃;

F) under the effect of shortening system, the nitro of compound VI I is reduced, and gets compound VIII, and the said catalystsystem of this step is: adopt Pd/C hydrogenation reaction reduction nitro; Solvent is methyl alcohol or ethanol;

G) under the alkaline condition, compound VIII and 4-iodo-4-deoxidation-4 '-demethyl epipodophyllotoxin reaction gets compound I X; Alkali is triethylamine and barium carbonate; Solvent is methylene dichloride, toluene, THF, acetonitrile, N, and dinethylformamide, methyl-sulphoxide etc. are preferably acetonitrile; This step temperature of reaction is 0-40 ℃, is preferably 20-30 ℃;

H) compound I X removes the protection base under acidic conditions, gets compound I-1; Said acid is hydrochloric acid, sulfuric acid, trifluoracetic acid etc., is preferably trifluoracetic acid; Solvent is methylene dichloride, ethylene dichloride, acetonitrile, THF etc., is preferably methylene dichloride.

Compound involved in the present invention has good antineoplastic activity, and the modes such as oral, intravenous injection and intramuscular injection of can carrying out are clinically used.

Embodiment

Novel Etoposide verivate of the present invention and preparation method be narration in more detail in following embodiment, but embodiment is not construed as limiting the invention.

Embodiment 1

1.1 the preparation of compound 2

5g 4-bromo-butyric acid is dissolved in the 100 mL methyl alcohol, and room temperature dripping thionyl chloride 3.3 mL dripped off continued reaction 1 hour, behind removal of solvent under reduced pressure and the excessive sulfur oxychloride colourless liquid 5.4 g.Productive rate 99.6%.

Embodiment 2

1.2 the preparation of compound 3

Under the room temperature, 4.52 g salt of wormwood are added the N that contains p-NP 3.79 g, in the dinethylformamide solution, stirring adds 4-bromo-butyric acid methyl esters 5.4 g after half a hour, and 100 ℃ were reacted 12 hours.After being cooled to room temperature, add 100 mL water, with 100 mL ethyl acetate extractions, organic phase is with washing, and saturated common salt is washed, and anhydrous sodium sulfate drying filters, and underpressure distillation gets yellow oil.Ether is added in this oily matter, separate out a large amount of solids, filter the back, get light yellow solid 3.3 g, productive rate 46.0% after the drying with a small amount of ether washing.

Embodiment 3

1.3 the preparation of compound 4

Under the room temperature, 2.1 g are dissolved in the 10 mL THFs with 4-(4-nitrophenoxy) methyl-butyrate, add 3M aqueous sodium hydroxide solution 5 mL, and the 2 hours postcooling that reflux are to room temperature.Add 1M hydrochloric acid 20 mL, use ethyl acetate extraction, organic phase is with washing, and saturated common salt is washed, and anhydrous sodium sulfate drying filters, and underpressure distillation gets yellow solid 1.9 g, productive rate 96.0%.

Embodiment 4

1.4 the preparation of compound 5

450 mg are dissolved in 5 mL toluene with 4-(4-nitrophenoxy) butyric acid, and dripping thionyl chloride 440 μ l under the room temperature are warming up to 60 ℃ of reactions 2 hours, are cooled to revolve after the room temperature to steam to remove to desolvate.

Embodiment 5

1.5 the preparation of compound 6

With 988 mg TBDMSO-NH-boc, 1.67 mL triethylamines, 24 mg N, TMSDMA N dimethylamine yl pyridines are dissolved in 10 mL acetonitriles and are cooled to-10 ℃, drip to be dissolved with compound 5Acetonitrile solution, react after 10 minutes and to add proper silica gel and mix appearance, be the eluent column chromatography with the ethyl acetate petroleum ether, white solid 434 mg, productive rate 46%.

¹H?NMR?(CDCl ₃,?500?MHz):?0.15?(s,?6H),?1.01?(s,?9H),?1.58?(s,?9H),?2.18-2.22?(m,?2H),?3.03?(t,? J?=?7.0?Hz,?2H),?4.14?(t,? J?=?6.2?Hz,?2H),?6.95?(d,? J?=?9.2?Hz,?2H),?8.20?(d,? J?=?9.1Hz,?2H)。

Embodiment 6

1.6 the preparation of compound 7

With compound 6400 mg are dissolved in the 20 mL ethanol, add 40 mg palladium carbon (10%), hydrogenation reaction under the room temperature.2 termination reactions as a child remove by filter catalyzer, and filtrating is revolved dried product 360 mg, productive rate 98%.

¹H?NMR?(CDCl ₃,?500?MHz):?0.14?(s,?6H),?0.99?(s,?9H),?1.54?(s,?9H),?2.05-2.15?(m,?2H),?2.97?(t,? J?=?10?Hz,?2H),?3.25-3.50?(br,?2H),?3.93?(t,? J?=?5?Hz,?2H),?6.61?(d,? J?=?10?Hz,?2H),?6.71?(d,? J?=?10Hz,?2H)。

Embodiment 7

1.7 the preparation of compound 8

With 9-(4,6-O-ethylidene-β-D-glucopyranoside)-4'-demethyl epipodophyllotoxin 322 mg, TMS-Cl 239 mg, NaI 330 mg react 2h in 15 mL acetonitriles after, add TEA 195 mg, BaCO ₃325 mg stir and add compound after 10 minutes 7300 mg, stirred overnight at room temperature.Reaction solution through filter concentrate after, get said product through column chromatography for separation.

¹H?NMR?(CDCl ₃,?500?MHz):?0.17?(s,?6H),?1.02?(s,?9H),?1.69?(s,?9H),?2.10-2.20?(m,?2H),?2.95-3.20?(m,?4H),?3.60?(s,?1H),?3.81?(s,?6H),?3.95-4.70?(m,?6H),?5.47?(s,?1H),?5.97?(d,? J?=?9.5?Hz,?2H),?6.35?(s,?2H),?6.48-6.82?(m,?6H)。

Embodiment 8

1.8 the preparation of compound 9

With compound 850 mg are dissolved in the 2 mL methylene dichloride, add the acid of 1 mL, three paraldehydes, room temperature reaction 16 hours.Column chromatography for separation behind the dried solvent is revolved in decompression, compound 920 mg. ¹H?NMR?(CDCl ₃,?500?MHz):1.95-2.10?(m,?2H),?2.27-2.31?(m,?2H),?3.05-3.30?(m,?2H),?3.61-3.62?(m,?1H),?3.64?(s,?6H),?3.90-4.00?(m,?3H),?4.30-4.95?(m,?4H),?5.97?(d,? J?=?5.6?Hz,?2H),?6.40?(s,?2H),?6.52?(s,?1H),?6.68-6.80?(m,?5H);?HRMS?(ESI):?m/z?calcd?for?C ₃₁H ₃₂N ₂NaO ₁₀?[M+Na] ⁺?626.1949,?found?615.1934。

Embodiment 9 HDAC suppress active testing:

With Ac-Lys-Tyr-Lys (Ac)-AMC is substrate, adopts fluorescence detection, in the flat microwell plate in 96 holes or 384 holes, detects enzymic activity.Substrate A c-Lys-Tyr-Lys (Ac)-AMC is respectively through HDAC1, and behind the HDAC3 deacetylation, the product A MC that utilizes pancreatin hydrolysis to obtain excites under the 460nm emission light at the 355nm of fluorescence detector and can be detected fluorescent signal.Through detecting the variation of fluorescent signal in time, calculate initial velocity of reaction.

With Boc-lys (Ac)-AMC is substrate, adopts fluorescence detection, in the flat microwell plate in 96 holes or 384 holes, detects enzymic activity.Substrate B oc-lys (Ac)-AMC is behind the HDAC6 deacetylation, and the product A MC that utilizes pancreatin hydrolysis to obtain excites under the 460 nm emission light at 355 nm of fluorescence detector and can be detected fluorescent signal.Through detecting the variation of fluorescent signal in time, calculate initial velocity of reaction.

The inhibition of table 1. couple HDAC is active

Table 1 has shown that compound of the present invention has the activity suitable with SAHA to HDAC, and compound ZX-010 wherein, ZX-011 demonstrate than the better HDAC of SAHA and suppress active.

Embodiment 10The test of the cytotoxic activity of all compounds is all carried out on corresponding tumor cell line, and 6000～10000 tumour cells are arranged in the nutrient solution of per 100 μ L, place 96 orifice plates (Falcon, CA).Tumour cell is divided into three parts, with the drug-treated of gradient concentration, cultivates 72h down at 37 ℃, adopts the srb assay test.Drug concentrations IC50 used amount effect curve to calculate when 50% growth of tumour cell suppressed.

The GIA of table 2. pair HCT116 cell

Table 2 has shown that compound of the present invention has good inhibition activity to HCT116.

Claims

1. epipodophyllotoxin derivatives that contains the hydroxamic acid structure and the pharmacy acceptable salt thereof shown in the general formula I:

Wherein,

X is-NH-,-O-,-S-,-NH-CH ₂-,-O-CH ₂-or-S-CH ₂-;

Y is aromatic nucleus, substituted aromatic nucleus, fragrant heterocycle or substituted fragrant heterocycle;

Z is-O-,-NHCO-or-CONH-;

B is the alkyl of C1-C8, alkenyl, alkynyl or the naphthenic base of C2-C8.

2. the epipodophyllotoxin derivatives that contains the hydroxamic acid structure shown in the said general formula I of claim 1 and the preparation method of pharmacy acceptable salt thereof, this method comprises the steps:

Wherein, n=3-7;

A) compound I I reacts with sulfur oxychloride in methyl alcohol, gets compound III, and temperature of reaction is 0-40 ℃;

B) under the alkali action condition, compound III and nitrophenols reaction obtain compound IV;

C) under the alkali action condition, the compound IV hydrolysis gets compound V;

D) under anhydrous condition, compound V and sulfur oxychloride reaction get compound VI, and temperature of reaction is 0-80 ℃;

E) under alkali and catalyzer acting in conjunction, compound VI and N-boc-O-TBS azanol reaction get compound VI I, and temperature of reaction is-20-20 ℃;

F) under the effect of shortening system, the nitro of compound VI I is reduced, and gets compound VIII;

G) under the alkaline condition, compound VIII and 4-iodo-4-deoxidation-4 '-demethyl epipodophyllotoxin reaction gets compound I X, and temperature of reaction is 0-40 ℃;

H) compound I X removes the protection base under acidic conditions, gets compound I-1.

3. compsn that is used to treat cancer, said composition comprises described epipodophyllotoxin derivatives and pharmacy acceptable salt and the pharmaceutically acceptable carrier that contains the hydroxamic acid structure of one or more claims 1 of treating significant quantity.

4. described epipodophyllotoxin derivatives and the application of pharmacy acceptable salt in the preparation antitumor drug thereof that contains the hydroxamic acid structure of claim 1.

5. described epipodophyllotoxin derivatives and the application of pharmacy acceptable salt in preparation treatment colorectal carcinoma or lung cancer drugs thereof that contains the hydroxamic acid structure of claim 1.