CN1557814A

CN1557814A - Homocamptothecin compound and its preparation method and use

Info

Publication number: CN1557814A
Application number: CNA2004100162290A
Authority: CN
Inventors: 张万年; 杨松; 姚建忠; 余建鑫; 盛春泉; 朱杰; 宋云龙; 张珉; 缪震元; 张晶
Original assignee: Second Military Medical University SMMU
Current assignee: Second Military Medical University SMMU
Priority date: 2004-02-12
Filing date: 2004-02-12
Publication date: 2004-12-29
Anticipated expiration: 2024-02-12
Also published as: CN100408582C

Abstract

The invention relates to the technical field of medicine, and relates to a new homocamptothecin compound with antitumor activity, its application and a new preparation method of the homocamptothecin compound. The _structure of _the compound of the present invention is shown in general formula (I), R ₁ is a lower haloalkyl group, a new substituted heterocycloalkyl group or a _substituent containing a substituted amino group; Substituted by group or amino group; R ₅ represents lower alkyl. The preparation method of the homocamptothecin compounds of the invention has low cost and high yield. The compound of the invention has the function of inhibiting topoisomerase, has antitumor activity and antiviral activity, and can be used as a topoisomerase inhibitor and for preparing antitumor drugs and antiviral drugs.

Description

High camptothecine compounds and its production and use

Technical field

The present invention relates to medical technical field, is a kind of new high camptothecine compounds of anti-tumor activity and its production and use that has.

Background technology

(camtptothecin is a kind of natural alkaloid that obtains that extracts from camplotheca acuminata CPT) to camptothecine, has five rings phase structure also.Natural camptothecine structure is as follows:

Camptothecine shows antiproliferative activity to the various human tumor cell line, and its anti-tumor activity is realized by suppressing the DNA topoisomerase I.

The hexa-atomic Alpha-hydroxy lactonic ring E ring of camptothecine is easy to be hydrolyzed into the carboxylate form of non-activity in vivo, and its activity in vivo is reduced greatly.After its hexavalent Alpha-hydroxy lactonic ring structure of modification is become seven yuan of beta-hydroxy lactonic ring analogues, obtained new camptothecin analogues, be called high camptothecine (homocamptothecin, hCPT) compounds, show the enhanced anti-tumor activity, also had metabolic stability enhanced advantage.Structure is shown in general formula (I).

R wherein ₁Expression H, low alkyl group, (CH ₂) NR ₆R ₇(R wherein ₆And R ₇Represent H, low alkyl group or rudimentary aralkyl independently) or unsubstituted (CH ₂) [N=X] or the group (CH that replaced by low alkyl group ₂) [N=X], wherein [N=X] represents 4 to 7 yuan of heterocyclic radicals, X represents to constitute the needed chain of heterocyclic radical and is selected from CH, CH ₂, O, S, N or NR ₉, R ₉Expression low alkyl group or aralkyl.

R ₂, R ₃, R ₄Expression H, halogen, low alkyl group, (CH ₂) NR ₆R ₇, OR ₆, or R ₃And R ₄Common methylene-dioxy or ethylenedioxy, the R of forming ₆, R ₇The expression low alkyl group.

R ₅The expression ethyl.

The complete synthesis route of existing high camptothecine compounds: 1, people's such as Olivier Lavergne complete synthesis route, can be referring to J.Med.Chem.41:5410 (1998); 2, people's such as Curran complete synthesis route can be referring to J Med Chem., 42:3018 (1999), Angew Chem Int Ed Engl.34:2683-2684 (1995).These two lines need be used some reagent that cost an arm and a leg and be difficult for obtaining, as palladium, Me ₂NCH ₂CH ₂NMeCHO, Me ₃SnSnMe ₃Deng, cost is higher, and the total recovery of these two lines all is lower than 5%.

Summary of the invention

The invention provides the high high camptothecine compounds of new anti-tumor activity, and a kind of new high camptothecine compounds preparation method that cost is low, yield is high is provided.

The The compounds of this invention structure is shown in general formula (I), but the group difference that connects, and 1, R ₁For low-grade halogenated alkyl, new hydrophilic substituted heterocycle alkyl or contain the substituting group of substituted-amino; 2 or/and at R ₂, R ₃, R ₄The position is that nitro replaces; 3 or/and with method well known in the art (as methods such as iron powder reducing, hydro-reductions) with R ₂, R ₃, R ₄Can obtain NH behind the nitroreduction of position ₂The high camptothecine that replaces.

Specifically, theme of the present invention is any mixture or its pharmaceutical salts of the compound with general formula (I), its racemic modification, enantiomeric forms or these forms of brand new.Wherein

R ₁The expression low-grade halogenated alkyl, condition is R ₂, R ₃, R ₄Have at least one to be the group beyond the hydrogen, low-grade halogenated alkyl refers to the straight or branched alkyl that contains 1-6 carbon atom, wherein contains 1-3 halogen atom (fluorine, chlorine, bromine, iodine) and replaces, as CH ₂Cl, CH ₂Br, CH ₂I, CH ₂CH ₂Cl, CH ₂CF ₃Deng; The C that preferably contains 1 halogen atom _1-3Alkyl, particularly preferably be chloromethyl;

Or R ₁Expression (CH ₂) _m[N=X], wherein [N=X] represents 4 to 7 yuan of heterocyclic radicals, X represents to constitute the needed chain of heterocyclic radical and is selected from CH, CH ₂, O, S, N or NR ₉, m represents the integer between the 0-6.Unlike the prior art be R here ₉What represent is aryl or heterocyclic radical.Aryl refers to and replaces or unsubstituted monocycle or dicyclic compound such as phenyl, naphthyl etc., the preferably phenyl that contains at least one aromatic ring; Heterocyclic radical refers to and replaces or the unsubstituted heterocyclic group that contains 1 to 3 heteroatoms (as N, S, P, O etc.) such as piperazinyl, piperidyl, pyridyl, morpholinyl etc., preferably contains a heteroatomic heterocyclic group, and particularly preferred is pyridyl;

And/or at R ₂, R ₃, R ₄In have one at least for nitro or when amino, R ₁Expression H, low alkyl group, (CH ₂) _mNR ₆R ₇, R ₆And R ₇Represent H, low alkyl group or rudimentary aralkyl independently, m represents the integer between the 0-6;

R ₂, R ₃, R ₄Can be identical or different, represent following groups: H, halogen, low alkyl group, OR respectively independently ₈(R ₈The expression low alkyl group), nitro, amino, perhaps R ₃And R ₄Form 3 or 4 yuan chain together, perhaps R ₂And R ₃Form 3 or 4 yuan chain together, wherein the element of this chain is selected from CH ₂, O, as methylene-dioxy or ethylenedioxy;

R ₅The expression low alkyl group, preferable methyl, ethyl, propyl group; Particularly preferably be ethyl.

In this article, described low alkyl group refers to the straight or branched alkyl that contains 1 to 6 carbon atom, as methyl, ethyl, sec.-propyl etc.Low-grade halogenated alkyl refers to the straight or branched alkyl that contains 1 to 6 carbon atom that 1 to 3 halogen atom replaces.Rudimentary aralkyl represents to be connected with the low alkyl group of aryl.Halogen refers to fluorine, chlorine, bromine, iodine.

Contain an asymmetric c atom in the beta-hydroxy lactonic ring of The compounds of this invention, two kinds of configurations of R and S are arranged.Its racemic modification can be by splitting method [as the Split Method of Olivier Lavergne etc., J.Med.Chem.41:5410 (1998)] obtain optically pure enantiomorph.The compounds of this invention comprises these two kinds of enantiomorph configurations and their various combinations, and racemic compound.

The present invention also provides a kind of method of the brand-new high camptothecine compounds of preparation.Synthetic route is as follows

But starting material compound A reference literature similar approach obtains [J.Med.Chem.23:554 (1980) such as Wani; J.Med.Chem.29:1553 such as Wall (1986)].

1, with general formula A compound with reductive agent reduction lactonic ring carbonyl after, melt ring with oxidizer oxygen, the Formula B compound,

Wherein n is 1 or 2, R ₅Expression contains the low alkyl group of 1-6 carbon atom, and preferred reductive agent is POTASSIUM BOROHYDRIDE or sodium borohydride, and preferred oxygenant is a periodate, particularly preferably is sodium periodate and potassium periodate.

2, with Formula B compound and R ₁₂R ₁₃YCCOOR ₁₁Represented alpha-halogen acid esters obtains general formula C compound by thunder formal thatch base well known in the art (Reformatsky) reaction,

R wherein ₁₁Be lower straight or branched-chain alkyl, R ₁₂, R ₁₃Represent H or low alkyl group respectively, Y is a halogen, and n is 1 or 2, R ₅For containing the low alkyl group of 1～6 carbon atom.

3, general formula C compound is hydrolyzed and cyclization under the condition that acid exists, obtains midbody compound D, acid here comprises organic acid and mineral acid, and preferred acid is hydrochloric acid, sulfuric acid, trifluoroacetic acid.

4, midbody compound D is obtained formula (I) structural compounds with adjacent amino aromatic aldehyde, ketone (general formula E compound) coupling of different replacements

Wherein, R ₁Groups such as expression H, low alkyl group, low-grade halogenated alkyl, aryl, rudimentary aralkyl, cycloalkyl, rudimentary hydroxyalkyl; R ₂, R ₃, R ₄Represent H, halogen, low alkyl group, OR independently ₈, nitro, perhaps R ₃And R ₄Form 3 or 4 yuan chain together, perhaps R ₂And R ₃Form 3 or 4 yuan chain together, the element of this chain is selected from CH ₂, O, N, S; R ₅And R ₈The expression low alkyl group, rudimentary hydroxyalkyl represents to have the low alkyl group of hydroxyl replacement.

The different adjacent amino aromatic ketones that replace can be according to literature method [J.Org.Chem 44:578 (1979) such as Sugasawa; J.Am.Chem.Soc.100:4842 such as Sugasawa (1978)] obtain: difference is replaced (R ₂, R ₃, R ₄) aniline and different nitrile (R ₁CN) at BCl ₃With other Lewis acid (as AlCl ₃, TiCl ₄) existence that waits down reaction obtain containing the general formula E compound of different substituents.

Below with 1,1-ethylenedioxy-5-O-(5 '-ethyl-5 '-hydroxyl-2 ' H, 5 ' H, 6 ' H-6-oxygen pyrans) [3 ', 4 '-f] Δ ^{6 (8)}Tetrahydrochysene indolizine (compound K, i.e. n=1, R ₅General formula A compound during for ethyl) for example synthetic route is done detailed description:

But starting material compound K reference literature method obtains [J.Med.Chem.23:554 (1980) such as Wani with 38% yield; J.Med.Chem.29:1553 such as Wall (1986)], do not repeat them here.

Concrete synthesis step is as follows:

1, at first, the structure of the company's of obtaining glycol was carried out oxidation with it with suitable oxygenant again after compound K reduced with appropriate reductant, obtained compound L.Here preferred reductive agent is POTASSIUM BOROHYDRIDE or sodium borohydride, and preferred oxygenant is a periodate, and wherein sodium periodate and potassium periodate are particularly preferred.Solvent for use is polar solvent such as water, alcohol, carboxylic acid, dimethyl sulfoxide (DMSO) etc. or its mixed solvent, and preferably water, alcohol, carboxylic acid or its mixed solvent particularly preferably are water, methyl alcohol, acetate or its mixed solvent.

2, with compound L and R ₁₂R ₁₃YCCOOR ₁₁Represented alpha-halogen acid esters reacts in organic solvent in the presence of metal or metal derivative, obtains compound M.R wherein ₁₁Expression contains the alkyl of the straight or branched of 1 to 6 carbon atom, is preferably methyl, ethyl, sec.-propyl, the tertiary butyl here, and wherein the tertiary butyl is particularly preferred.R ₁₂, R ₁₃Represent H or low alkyl group respectively.Y represents fluorine, chlorine, bromine, iodine.Here the preferred bromo acid esters of alpha-halogen acid esters, wherein, the alpha bromoisobutyric acid tert-butyl ester is particularly preferred.Here said metal is selected from Zn, Mg or Li etc., wherein preferred Zn and Mg, and Zn is particularly preferred.Organic solvent is selected from ether, benzene, tetrahydrofuran (THF), dimethoxy second (first) alkane, dimethyl sulfoxide (DMSO) etc., preferred tetrahydrofuran (THF) and glycol dimethyl ether, and wherein tetrahydrofuran (THF) is particularly preferred.

3, compound M is hydrolyzed and cyclization under the condition that acid exists, obtains compound N.Here acid comprises organic acid and mineral acid, and preferred acid is hydrochloric acid, sulfuric acid, trifluoroacetic acid, and wherein trifluoroacetic acid is particularly preferred.

4, compound N and E are reacted under the condition that acid exists, obtain containing the high camptothecine compounds (II) of seven yuan of beta-hydroxy lactonic rings.Here said acid comprises organic acid (as tosic acid, acetate, formic acid etc.) and mineral acid (example hydrochloric acid, sulfuric acid etc.), organic acid preferably, and wherein tosic acid is particularly preferred.Solvent can be selected from benzene, toluene, ethanol, acetate isopolarity or non-polar solvent, non-polar solvents such as preferred benzene or toluene, and particularly preferred is toluene.

Can reach 85% by compound K to the yield of intermediate N, the total recovery of compound N can reach 32%.

The yield of intermediate N and the coupling of general formula E compound is between 40%～80%, and in minimum 40%, the total recovery of high Comptothecin compounds (II) surpasses 12%.

At R ₁During for chloromethyl, II can with heterocycle or the reactions of non-heterocyclic compound such as secondary amine such as piperidines, single substituted-piperazinyl, morpholine, dimethylamine, generate R ₁The high Comptothecin compounds of beta-hydroxy lactonic ring that contains groups such as heterocycle, aliphatic amide.

Will be clear that method provided by the invention can also prepare the high camptothecine compounds that The compounds of this invention other groups in addition replace, and comprises high camptothecine compounds more of the prior art, R ₁Can be H, low alkyl group, aryl, rudimentary aralkyl, cycloalkyl, rudimentary hydroxyalkyl etc., as: intermediate N and adjacent amino aromatic aldehyde or the reaction of its Glycol Acetal can obtain R ₁High camptothecine compounds for H.

Some compound of the present invention can be prepared as the form of pharmaceutical salts according to ordinary method.Comprise its inorganic acid salt and organic acid salt: mineral acid includes, but is not limited to hydrochloric acid, sulfuric acid, phosphoric acid, bisphosphate, Hydrogen bromide, nitric acid etc., and organic acid includes, but is not limited to acetate, toxilic acid, fumaric acid, tartrate, succsinic acid, lactic acid, tosic acid, Whitfield's ointment, oxalic acid or the like.

Formula I compound of the present invention has the effect that suppresses topoisomerase, and has anti-tumor activity.Prior art hints that compound of the present invention has antiviral activity [Chiang J.Li etc., The Journal of Biological Chemistry, 269:7051 (1994)], and therefore compound of the present invention can be used for preparing the corresponding treatment medicine.

Compound of the present invention has anti-tumor activity, they can be used for treating tumour, comprise the cancer that positions such as esophagus, stomach, intestines, rectum, oral cavity, pharynx, larynx, lung, colon, mammary gland, uterus, uterine endometrium, ovary, prostate gland, testis, bladder, kidney, liver, pancreas, bone, reticular tissue, skin, eye, brain and central nervous system take place, and thyroid carcinoma, leukemia, Hokdkin disease, lymphoma and myelomatosis etc.

Compound of the present invention can also be used for the treatment of virus infection.

The pharmacologically active of The compounds of this invention makes it can be used to prepare antitumor drug or antiviral, so the present invention also comprises with these compounds and pharmaceutical salts thereof the pharmaceutical composition as activeconstituents.This pharmaceutical composition can be a solid form, also can be liquid form.

The present invention also comprises the purposes of formula I compound aspect the following medicine of preparation: the medicine that suppresses topoisomerase; Antitumor drug and antiviral aspect.

Embodiment

Below in conjunction with embodiment the present invention is described in detail, but the following example should not regarded limitation of the scope of the invention as.

Embodiment 1 preparation compound N

A, 1,1-ethylenedioxy-6-methanoyl methyl-7-propionyl-5-oxygen-1,2,3,5-tetrahydrochysene indolizine (compound L) synthetic

Reference literature method (J.Med.Chem.23:554 (1980) such as Wani; J.Med.Chem.29:1553 such as Wall (1986)) obtain starting raw material 1,1-ethylenedioxy-5-O-(5 '-ethyl-5 '-hydroxyl-2 ' H, 5 ' H, 6 ' H-6-oxygen pyrans) [3 ', 4 '-f] Δ ^{6 (8)}Tetrahydrochysene indolizine (compound K).Getting the 8g compound K is dissolved in the 170ml methyl alcohol, add the 0.96g POTASSIUM BOROHYDRIDE, heated and stirred is after 20 minutes in 50 ℃ of oil baths, add the 120ml acetate and the 30ml aqueous solution that contains the 7.2g sodium periodate, continue to stir after 20 minutes in the impouring 600ml water, after extracting with methylene dichloride 100ml * 5, drying, solvent evaporated in vacuo gets compound L 7.5g.With the tetrahydrofuran (THF) recrystallization, mp:126-7 ℃.

¹HNMR(CDCl ₃)：1.19(t，3H)，2.41(t，2H)，2.8(q，2H)，4.1(m，6H)，5.2(s，2H)，6.2(s，1H)，8.0(s，1H)。

Synthesizing of B, 3-hydroxyl-3-(6-methylol-1,1-ethylenedioxy-5-oxygen-1,2,3,5-tetrahydrochysene indolizine-7-yl)-valeric acid tert-butyl ester (compound M)

In the 250ml three-necked bottle, under the nitrogen atmosphere, add the 120ml tetrahydrofuran (THF), the 9gZn powder is put 70 ℃ of oil baths and is heated to backflow, drips the 20ml bromo-acetic acid tert-butyl, drip the 50ml tetrahydrofuran solution that contains the 5.5g compound L then, continue to stir after 20 minutes, stopped reaction adds the 100ml saturated aqueous ammonium chloride, extract with methylene dichloride 100ml * 3, solvent evaporated is crossed silicagel column, gets compound M6.9g.

¹HNMR(CDCl ₃)：0.87(t，3H)，1.35(s，9H)，1.88(m，2H)，2.37(t，2H)，2.80(dd，2H)，4.1(m，6H)，4.3(s，1H)，4.79(s，1H)，4.9(dd，2H)，6.4(s，1H)。

MS：m/z?395

C, 9-ethyl-9-hydroxyl-2,3,8,9-tetrahydrochysene-5H-6-oxa--3a-azepine-ring indenes in heptan-1,4,7-triketone (compound N) synthetic

Method 1: synthetic compound N under the mineral acid condition

In the 25ml flask, add the saturated methylene dichloride 10ml of HCl, acetone 3ml, add 220mg compound M again, after stirring is spent the night in room temperature, add the 2ml concentrated hydrochloric acid, continue reaction 10 hours, transfer pH4-5, solvent evaporated with dilute sodium bicarbonate solution, with acetone extraction, evaporate to dryness again with 50% aqueous acetone solution recrystallization, gets yellow crystals, drying gets compound N 50mg.

Method 2: synthetic compound N under the organic acid condition

Get 4.6g compound M and add the 28ml trifluoroacetic acid, in stirring at room 10 hours, solvent evaporated, with 50% aqueous acetone solution recrystallization, compound N 3.04g, yellow crystals.mp：121-123℃。

¹HNMR (deuterated acetone): 0.9 (t, 3H), 1.78-1.88 (m, 2H), 2.96 (t, 2H), 3.22-3.4 (ABq, 2H), 4.25 (t, 2H), 5.38-5.51 (ABq, 2H), 7.0 (s, 1H).

MS (EI): calculated value 477, measured value 477.

Embodiment 2

The preparation 7-chloromethyl-high camptothecine of 10-chlorine (general formula I I compound, wherein R ₁, R ₂, R ₃, R ₄Be followed successively by chloromethyl, H, Cl, H) A, 2 '-amino-2,4 '-dichloroacetophenone synthetic

Two mouthfuls of bottles of 25ml, the BCl of adding 3ml1M ₃Hexane solution is put the ice bath cooling down, drips the 3ml benzole soln of 0.32g p-Chlorobenzoic acid amide, remove ice bath after adding, add 0.19ml chloromethyl cyanide and 0.37g aluminum trichloride (anhydrous), logical nitrogen reflux 6 hours, remove oil bath, put and cool off adding 3ml2N hydrochloric acid down in the ice bath, reheat, extract with methylene dichloride 8ml * 3 to the solid dissolving, organic layer is washed with 20ml, evaporate to dryness behind the anhydrous magnesium sulfate drying gets 2 '-amino-2,4 '-dichloroacetophenone 0.2g.

Synthesizing of B, 7-chloromethyl-high camptothecine of 10-chlorine

Get 45mg2 '-amino-2,4 '-dichloroacetophenone, 60mg9-ethyl-9-hydroxyl-2,3,8,9-tetrahydrochysene-5H-6-oxa--3a-azepine-ring indenes in heptan-1,4,7-triketone (compound N) is in 40ml toluene, reflux water-dividing after 30 minutes, adds the 10mg tosic acid, continue to reflux 5 hours, stop heating, cooling back suction filtration, filter cake is washed with acetone, gets yellow product 7-chloromethyl-high camptothecine 75mg of 10-chlorine.

¹HNMR(DMSO)：0.86(t，3H)，1.85(q，2H)，3.04-3.51(ABq，2H)，5.38-5.56(m，6H)，6.05(s，1H)，7.41(s，1H)7.94(d，1H)，8.23(d，1H)，8.50(s，1H)

Embodiment 3

The preparation 7-chloromethyl-high camptothecine of 10-fluorine (general formula I I compound, wherein R ₁, R ₂, R ₃, R ₄Be followed successively by chloromethyl, H, F, H) A, 2 '-amino-2-chloro-4 '-fluoro acetophenone

According to the method for embodiment 2A, replace p-Chlorobenzoic acid amide with para-fluoroaniline, obtain 2 '-amino-2-chloro-4 '-fluoro acetophenone.

B, the 7-chloromethyl-high camptothecine of 10-fluorine

According to the method for embodiment 2B, replace 2 '-amino-2 with 2 '-amino-2-chloro-4 '-fluoro acetophenone, 4 '-dichloroacetophenone gets 7-chloromethyl-high camptothecine of 10-fluorine, is yellow solid.

¹HNMR(DMSO)：0.919(t，3H)，1.9(q，2H)，3.11-3.50(Abq，2H)，5.42-5.59(m，6H)，6.0(s，1H)，7.45(s，1H)7.86(m，1H)，8.21(m，1H)，8.32(m，1H)

Embodiment 4

The preparation 7-chloromethyl-high camptothecine of 10-bromine (general formula I I compound, wherein R ₁, R ₂, R ₃, R ₄Be followed successively by chloromethyl, H, Br, H)

Method according to A, B among the embodiment 2 describe replaces p-Chlorobenzoic acid amide with para-bromoaniline, gets yellow product 7-chloromethyl-high camptothecine of 10-bromine.

¹HNMR(DMSO)：0.86(t，3H)，1.86(q，2H)，3.07-3.46(ABq，2H)，5.39-5.55(m，6H)，5.96(s，1H)，7.42(s，1H)8.02(d，1H)，8.14(d，1H).8.63(s，1H)

Embodiment 5

The preparation 7-chloromethyl-high camptothecine of 10-methyl (general formula I I compound, wherein R ₁, R ₂, R ₃, R ₄Be followed successively by chloromethyl, H, CH ₃, H)

Method according to A, B among the embodiment 2 describe so that monomethylaniline is replaced p-Chlorobenzoic acid amide, gets yellow product 7-chloromethyl-high camptothecine of 10-methyl.

¹HNMR(DMSO)：0.88(t，3H)，1.86(q，2H)，2.50(s，3H)，3.07-3.46(ABq，2H)，5.35-5.55(m，6H)，7.39(s，1H)，7.74(d，1H)，8.1(d，1H)，8.18(s，1H)

Embodiment 6

The preparation 7-chloromethyl-high camptothecine of 10-methoxyl group (general formula I I compound, wherein R ₁, R ₂, R ₃, R ₄Be followed successively by chloromethyl, H, CH ₃O, H)

Method according to A, B among the embodiment 2 describe replaces p-Chlorobenzoic acid amide with P-nethoxyaniline, gets yellow product 7-chloromethyl-high camptothecine of 10-methoxyl group.

¹HNMR(DMSO)：0.88(t，3H)，1.86(q，2H)，3.07-3.46(ABq，2H)，4.01(s，3H)，5.36-5.55(m，6H)7.35(s，1H)，7.58(dd，1H)，7.66(d，1H)，8.11(d，1H)

Embodiment 7

The preparation 7-chloromethyl-high camptothecine of 11-methyl (general formula I I compound, wherein R ₁, R ₂, R ₃, R ₄Be followed successively by chloromethyl, H, H, CH ₃)

Method according to A, B among the embodiment 2 describe replaces p-Chlorobenzoic acid amide with a monomethylaniline, gets yellow product 7-chloromethyl-high camptothecine of 11-methyl.

¹HNMR(DMSO)：0.88(t，3H)，1.86(q，2H)，2.50(s，3H)，3.07-3.46(ABq，2H)，5.35-5.55(m，6H)，5.96(s，1H)7.39(s，1H)，7.66(d，1H)，8.0(s，1H)，8.29(d，1H)

Embodiment 8

The preparation 7-chloromethyl-high camptothecine of 11-chlorine (general formula I I compound, wherein R ₁, R ₂, R ₃, R ₄Be followed successively by chloromethyl, H, H, Cl)

Method according to A, B among the embodiment 2 describe replaces p-Chlorobenzoic acid amide with m-chloro aniline, gets yellow product 7-chloromethyl-high camptothecine of 11-chlorine.

¹HNMR(DMSO)：0.88(t，3H)，1.86(q，2H)，3.08-3.47(ABq，2H)，5.37-5.55(m，6H)，5.99(s，1H)，7.42(s，1H)，7.84(dd，1H)，8.26(d，1H)，8.44(d，2H)

Embodiment 9

The preparation 7-chloromethyl-high camptothecine of 11-fluorine (general formula I I compound, wherein R ₁, R ₂, R ₃, R ₄Be followed successively by chloromethyl, H, H, F)

Method according to A, B among the embodiment 2 describe replaces p-Chlorobenzoic acid amide with m-fluoroaniline, gets yellow product 7-chloromethyl-high camptothecine of 11-fluorine.

¹HNMR(DMSO)：0.88(t，3H)，1.86(q，2H)，3.08-3.47(ABq，2H)，5.98(s，1H)，7.42(s，1H)，7.76(m，1H)，7.96(m，1H)，8.50(m，1H)

Embodiment 10

Preparation 7-chloromethyl-10, the high camptothecine of 11-difluoro (general formula I I compound, wherein R ₁, R ₂, R ₃, R ₄Be followed successively by chloromethyl, H, F, F)

According to the method that A, B among the embodiment 2 describe, with 3, the 4-difluoroaniline replaces p-Chlorobenzoic acid amide, gets yellow product 7-chloromethyl-10, the high camptothecine of 11-difluoro.

¹HNMR(DMSO)：0.89(t，3H)，1.89(q，2H)，3.08-3.46(ABq，2H)，5.33-5.55(m，6H)，6.(s，1H)7.41(s，1H)，8.23(m，1H)，8.48(m，1H)

Embodiment 11

Preparation 7-chloromethyl-10, the high camptothecine of 11-dichloro (general formula I I compound, wherein R ₁, R ₂, R ₃, R ₄Be followed successively by chloromethyl, H, Cl, Cl)

According to the method that A, B among the embodiment 2 describe, with 3, the 4-dichlorphenamide bulk powder replaces p-Chlorobenzoic acid amide, gets yellow product 7-chloromethyl-10, the high camptothecine of 11-dichloro.

¹HNMR(DMSO)：0.87(t，3H)，1.84(q，2H)，3.07-3.46(ABq，2H)，5.38-5.60(m，6H)，5.98(s，1H)，7.41(s，1H)，8.47(s，1H)，8.70(s，1H)

Embodiment 12

Preparation 7-chloromethyl-10, the high camptothecine of 11-dimethyl (general formula I I compound, wherein R ₁, R ₂, R ₃, R ₄Be followed successively by chloromethyl, H, CH ₃, CH ₃)

According to the method that A, B among the embodiment 2 describe, with 3, the 4-xylidine replaces p-Chlorobenzoic acid amide, gets yellow product 7-chloromethyl-10, the high camptothecine of 11-dimethyl.

¹HNMR(DMSO)：0.88(t，3H)，1.86(q，2H)，2.5(m，6H)，3.07-3.46(ABq，2H)，5.33-5.55(m，3H)，7.37(s，1H)，7.97(s，1H)，8.15(s，1H)

Embodiment 13

Preparation 7-chloromethyl-9, the high camptothecine of 11-dichloro (general formula I I compound, wherein R ₁, R ₂, R ₃, R ₄Be followed successively by chloromethyl, Cl, H, Cl)

According to the method that A, B among the embodiment 2 describe, with 3, the 5-dichlorphenamide bulk powder replaces p-Chlorobenzoic acid amide, gets yellow product 7-chloromethyl-9, the high camptothecine of 11-dichloro.

¹HNMR(DMSO)：0.88(t，3H)，1.86(m，2H)，3.09-3.47(ABq，2H)，5.39-5.63(m，6H)，5.996(s，1H)，7.41(s，1H)，8.04(s，1H)，8.28(s，1H)

Embodiment 14

Preparation 7-chloromethyl-9, the high camptothecine of 11-dimethyl (general formula I I compound, wherein R ₁, R ₂, R ₃, R ₄Be followed successively by chloromethyl, CH ₃, H, CH ₃)

According to the method that A, B among the embodiment 2 describe, with 3, the 5-xylidine replaces p-Chlorobenzoic acid amide, gets yellow product 7-chloromethyl-9, the high camptothecine of 11-dimethyl.

¹HNMR(DMSO)：0.89(t，3H)，1.86(q，2H)，2.50(s，3H)，3.06(s，3H)，3.07-3.47(ABq，2H)，5.38-5.56(m，6H)，5.96(s，1H)，7.37(s，1H)，7.45(s，1H)，7.86(s，1H).

Embodiment 15

The preparation 7-chloromethyl-high camptothecine of 10-methyl isophthalic acid 1-fluorine (general formula I I compound, wherein R ₁, R ₂, R ₃, R ₄Be followed successively by chloromethyl, H, CH ₃, F)

Method according to A, B among the embodiment 2 describe replaces p-Chlorobenzoic acid amide with 3-fluoro-4-monomethylaniline, gets yellow product 7-chloromethyl-high camptothecine of 10-methyl isophthalic acid 1-fluorine.

¹HNMR(DMSO)：0.86(t，3H)，1.86(q，2H)，2.50(s，3H)，3.06-3.46(ABq，2H)，5.31-5.54(m，6H)，5.97(s，1H)，7.4(s，1H).7.9(d，1H)，8.35(d，1H).

Embodiment 16

The preparation 7-chloromethyl-high camptothecine of 10-methyl isophthalic acid 1-chlorine (general formula I I compound, wherein R ₁, R ₂, R ₃, R ₄Be followed successively by chloromethyl, H, CH ₃, Cl)

Method according to A, B among the embodiment 2 describe replaces p-Chlorobenzoic acid amide with 3-chloro-4-monomethylaniline, gets yellow product 7-chloromethyl-high camptothecine of 10-methyl isophthalic acid 1-chlorine.

¹HNMR(DMSO)：0.86(t，3H)1.86(q，2H)，2.60(s，3H)，3.06-3.46(ABq，2H)，5.36-5.56(m，6H)5.97(s，1H)，7.39(s，1H)，8.26(s，1H)，8.4(s，1H)

Embodiment 17

The preparation 7-chloromethyl-high camptothecine of 10-fluoro-11-chlorine (general formula I I compound, wherein R ₁, R ₂, R ₃, R ₄Be followed successively by chloromethyl, H, F, Cl)

Method according to A, B among the embodiment 2 describe replaces p-Chlorobenzoic acid amide with 3-chloro-4-fluoroaniline, gets yellow product 7-chloromethyl-high camptothecine of 10-fluorine 11-chlorine.

¹HNMR(DMSO)：0.86(t，3H)，1.85(q，2H)，3.08-3.46(ABq，2H)，5.38-5.55(m，6H)，7.40(s，1H)，8.41-8?48(m，2H)

Embodiment 18

The preparation 7-chloromethyl-high camptothecine of 10-methyl isophthalic acid 1-nitro (general formula I I compound, wherein R ₁, R ₂, R ₃, R ₄Be followed successively by chloromethyl, H, CH ₃, NO ₂)

Method according to A, B among the embodiment 2 describe replaces p-Chlorobenzoic acid amide with 3-nitro-4-methyl aniline, gets yellow product 7-chloromethyl-high camptothecine of 10-methyl isophthalic acid 1-nitro.

¹HNMR(DMSO)：0.88(t，3H)，1.86(q，2H)，2.74(s，3H)，3.08-3.47(ABq，2H)，5.39-5.56(m，6H)，5.99(s，1H)，7.44(s，1H)，8.50(s，1H)，8.75(s，1H).

Embodiment 19

Preparation 7-chloromethyl-9, the high camptothecine of 10-ethylenedioxy (general formula I I compound, wherein R ₁, R ₄Be followed successively by chloromethyl, H, R ₂And R ₃Common formation-OCH ₂CH ₂O-) and 7-chloromethyl-10, the high camptothecine of 11-ethylenedioxy (general formula I I compound, wherein R ₁, R ₂Be followed successively by chloromethyl, H, R ₃And R ₄Common formation-OCH ₂CH ₂O-)

A, 2-chloracetyl-4,5-ethylenedioxy aniline and 2-chloracetyl-3,4-ethylenedioxy aniline synthetic

The 50ml three-necked bottle adds 1MBCl ₃Hexane solution 10ml; dropping contains 3; the benzene liquid 10ml of 4-ethylenedioxy aniline 1.4ml; add 0.8ml chloromethyl cyanide and 1.3g aluminum trichloride (anhydrous) then; put the oil bath backflow after 12 hours; remove oil bath; add 1 water 0ml and 2N hydrochloric acid 7ml, stir after 15 minutes and divide 6 extractions, washing with methylene dichloride 80ml; anhydrous sodium sulfate drying; solvent evaporated, silica gel column chromatography separates obtaining 2-chloracetyl-4; pure product 0.9g of 5-ethylenedioxy aniline and 2-chloracetyl-3, the pure product 0.17g of 4-ethylenedioxy aniline.

¹HNMR：

2-chloracetyl-3,4-ethylenedioxy aniline (CDCl ₃): 4.23 (m, 2H), 4.35 (m, 2H), 4.75 (s, 2H), 6.22 (d, 1H), 6.87 (d, 1H).

2-chloracetyl-4,5-ethylenedioxy aniline (DMSO+CDCl ₃): 4.20 (m, 2H), 4.24 (m, 2H), 4.53 (s, 2H), 6.33 (s, 1H), 7.14 (s, 1H).

B, 7-chloromethyl-9, the high camptothecine of 10-ethylenedioxy synthetic

Get 2-chloracetyl-3,4-ethylenedioxy aniline 50mg, the compound N of 55mg embodiment 1 gained, 30ml toluene; put the oil bath reflux water-dividing, after 20 minutes, add the 10mg tosic acid, after continuing to reflux 20 minutes; solvent evaporated is crossed silicagel column, gets yellow product 75mg.

¹HNMR(DMSO)：0.88(t，3H)，1.86(q，2H)，3.06-3.46(ABq，2H)，4.7(m，4H)，5.35-5.54(m，6H)，5.94(s，1H)，7.34(s，1H)，7.51(d，1H)，7?75(s，1H).

C, 7-chloromethyl-10, the high camptothecine of 11-ethylenedioxy synthetic

Get 249mg2-chloracetyl-4,5-ethylenedioxy aniline, the compound N of 280mg embodiment 1 gained; 85ml toluene is put the oil bath reflux water-dividing, after 15 minutes; add the 30mg tosic acid, continue to reflux 50 minutes, inclining toluene; evaporated under reduced pressure adds 5ml acetone, grinds; suction filtration; get pure product 7-chloromethyl-10, the high camptothecine 361mg of 11-ethylenedioxy is the aureus solid.

¹HNMR(DMSO)：0.88(t，3H)，1.86(q，2H)，3.06-3.46(ABq，2H)，4.45(s，4H)，5.28-5.54(m，6H)，7.33(s，1H)，7.57(s，1H)，7.75(s，1H)

Embodiment 20

Preparation 7-(4-(3-trifluoromethyl) piperazine-1-ylmethyl)-10, the high camptothecine of 11-ethylenedioxy (general formula I I compound, wherein R ₁Be 4-(3-trifluoromethyl) piperazine-1-ylmethyl, R ₂Be H, R ₃And R ₄Common formation-OCH ₂CH ₂O-) and hydrochloride

A, 7-(4-(3-trifluoromethyl) piperazine-1-ylmethyl)-10, the high camptothecine of 11-ethylenedioxy synthetic

The 7-chloromethyl-10 that 30mg embodiment 19 obtains, the high camptothecine of 11-ethylenedioxy, stirring is dissolved in 3ml dimethyl sulfoxide (DMSO) (DMSO), adds the 3ml toluene liquid that contains 0.3g 1-(3-trifluoromethyl) piperazine, room temperature reaction 14 hours, the pressure reducing and steaming solvent gets brown solid, grinds with 2ml ethanol, suction filtration, get faint yellow solid, acetone is washed once, gets product 35mg.

¹HNMR(DMSO)：0.87(t，3H)，1.86(q，2H)，2.66(s，4H)，3.06(d，1H)，3.25(s，4H)，3.44(d，1H)，4.04(s，2H)，4.43(s，4H)，5.28(s，2H)，5.36-5.53(ABq，2H)，5.92(s，1H)，7.05(d，1H)，7.13(s，1H)，7.19(d，2H)，7.33(s，1H)，7.40(t，1H)，7.53(s，1H)，7.84(s，1H)。

B, preparation 7-(4-(3-trifluoromethyl) piperazine-1-ylmethyl)-10, the high camptothecine hydrochloride of 11-ethylenedioxy

Get 20mg7-(4-(3-trifluoromethyl) piperazine-1-ylmethyl)-10, the high camptothecine of 11-ethylenedioxy adds 10ml methyl alcohol, the 1ml concentrated hydrochloric acid, and stirring at room 2 hours, solvent evaporated gets its hydrochloride.

Embodiment 21

Preparation 7-(4-(3-chloro-5-trifluoromethyl-2-pyridyl) piperazine-1-ylmethyl)-10, the high camptothecine of 11-ethylenedioxy (general formula I I compound, wherein R ₁Be 4-(3-chloro-5-trifluoromethyl-2-pyridyl) piperazine-1-ylmethyl, R ₂Be H, R ₃And R ₄Common formation-OCH ₂CH ₂O-)

According to the method for embodiment 20, replace 1-(3-trifluoromethyl) piperazine with 1-(3-chloro-5-trifluoromethyl-2-pyridyl) piperazine, get 7-(4-(3-chloro-5-trifluoromethyl-2-pyridyl) piperazine-1-ylmethyl)-10, the high camptothecine of 11-ethylenedioxy.

¹HNMR(DMSO)：0.88(t，3H)，1.86(q，2H)，2.68(s，4H)，3.06-3.43(ABq，2H)，3.47(s，4H)，4.06(s，2H)，4.44(s，4H)，5.29(s，2H)，5.36-5.54(ABq，2H)，5.92(s，1H)，7.33(s，1H)，7.53(s，1H)，7.84(s，1H)，8.13(s，1H)，8.53(s，1H)。

Embodiment 22

Preparation 7-(4-(4-pyridyl) piperazine-1-ylmethyl)-10, (general formula I I compound, wherein R1 is 4-(4-pyridyl) piperazine-1-ylmethyl to the high camptothecine of 11-ethylenedioxy, R ₂Be H, R ₃And R ₄Common formation-OCH ₂CH ₂O-)

According to the method for embodiment 20, replace 1-(3-trifluoromethyl) piperazine with 1-(4-pyridyl) piperazine, get 7-(4-(4-pyridyl) piperazine-1-ylmethyl)-10, the high camptothecine of 11-ethylenedioxy.

¹HNMR(DMSO)：0.87(t，3H)，1.87(q，2H)，2.66(s，4H)，3.10(d，1H)，3.35(s，4H)，3.44(d，1H)，4.05s，2H)，4.43(s，4H)，5.29(s，2H)，5.35-5.53(ABq，2H)，5.92(s，1H)，6.83(d，2H)，7.33(s，1H)，7.53(s，1H)，7.85(s，1H)，8.15(s，2H)。

Embodiment 23

Preparation 7-(4-(4-acetyl phenyl) piperazine-1-ylmethyl)-10, the high camptothecine of 11-ethylenedioxy (general formula I I compound, wherein R ₁Be 4-(4-acetyl phenyl) piperazine-1-ylmethyl, R ₂Be H, R ₃And R ₄Common formation-OCH ₂CH ₂O-)

According to the method for embodiment 20, replace 1-(3-trifluoromethyl) piperazine with 1-(4-acetyl phenyl) piperazine, get 7-(4-(4-acetyl phenyl) piperazine-1-ylmethyl)-10, the high camptothecine of 11-ethylenedioxy.

¹HNMR(DMSO)：0.87(t，3H)，1.87(q，2H)，2.44(s，3H)，2.64(s，4H)，3.10(d，1H)，3.32(s，4H)，3.44(d，1H)，4.06(s，2H)，4.43(s，4H)，5.29(s，2H)，5.36-5.53(ABq，2H)，5.92(s，1H)，6.95(d，2H)，7.33(s，1H)，7.54(s，1H)，7.79(d，2H)，7.85(s，1H)。

The anti-tumor activity test of The compounds of this invention

Compound of the present invention has been carried out the tumor cell proliferation inhibition test.Test method adopts conventional mtt assay.

Cell strain is selected A549 (human lung carcinoma cell), LOVO (people's colon-cancer cell) for use.Nutrient solution is that DMEM+15%NBS+ is two anti-.

Sample liquid preparation: after DMSO (Merck) dissolving, adding PBS (-) is made into solution or the uniform suspension of 100 μ g/ml, with PBS (-) dilution that contains DMSO, ultimate density is respectively 10 μ g/ml, 1 μ g/ml, 0.1 μ g/ml, 0.01 μ g/ml, 0.001 μ g/ml, 0.0001 μ g/ml then.

The antitumor drug topotecan of will going on the market is made into reference substance solution with similarity condition.

It is 3 * 10 that the every hole of 96 orifice plates adds concentration ⁴The cell suspension 100 μ l of individual/ml, promptly 3000 cells/well are put 37 ℃, 5%CO ₂In the incubator.After 24 hours, add sample liquid and reference substance liquid respectively, 10 μ l/ holes, 37 ℃ act on 72 hours.Every hole adds MTT (3-(4,5-dimethylthiazole-2-yl)-2,5 phenylbenzene tetrazolium father-in-law bromides) the solution 20 μ l of 5mg/ml, act on and add lysate DMSO after 4 hours, put in the incubator in 100 μ l/ holes, and the moving microplate reader of the full order of inferior daily MK-2 is surveyed 570nm OD value.Calculation of half inhibitory concentration IC ₅₀

Test-results sees Table 1, and wherein, sample is meant the high camptothecine compounds for preparing among the corresponding embodiment, is illustrated in resulting high camptothecine compounds among the embodiment 3 as embodiment 3, in like manner analogizes.

Table 1 test compounds is to the half-inhibition concentration IC of tumour cell ₅₀(unit: μ g/ml)

Sample A549 LOVO

Embodiment 3 1.06 0.220

Embodiment 4 1.12 0.0340

Embodiment 6 0.324 0.0320

Embodiment 7 1.66 0.313

Embodiment 9 0.895 0.0343

Embodiment 14 1.57 0.0177

Embodiment 15 0.354 0.0334

Embodiment 17 0.393 0.0607

Embodiment 19B 0.0424 0.0025

Embodiment 20 0.821 0.162

Embodiment 21 4.17 0.431

Topotecan 2.47 0.0351

Above experimental result shows that compound of the present invention has good antineoplastic activity, and a plurality of compound activities are higher than the marketed drug topotecan, so The compounds of this invention and its esters can be used to prepare antitumor drug.

Claims

1. high camptothecine compounds comprises racemic form, enantiomeric forms and pharmaceutical salts thereof, and its chemical structure of general formula is suc as formula shown in (I),

It is characterized in that:

R ₁The expression low-grade halogenated alkyl, condition is R ₂, R ₃, R ₄Have at least one to be group beyond the hydrogen, described low-grade halogenated alkyl is to contain the straight or branched alkyl that contains 1-6 carbon atom that 1-3 halogen atom replaces; Or R ₁Expression (CH ₂) _m[N=X]; Or R ₁Expression H, low alkyl group, (CH ₂) _mNR ₆R ₇, condition is R ₂, R ₃, R ₄In have one at least for nitro or amino;

R ₂, R ₃, R ₄Represent H, halogen, low alkyl group, OR independently ₈, nitro, amino, perhaps R ₃And R ₄Form 3 or 4 yuan chain together, perhaps R ₂And R ₃Form 3 or 4 yuan chain together, wherein the element of this chain is selected from CH ₂, O;

R ₅The expression low alkyl group;

R ₆And R ₇Represent H, low alkyl group or rudimentary aralkyl independently, described rudimentary aralkyl is the low alkyl group that is connected with aryl;

R ₈The expression low alkyl group;

[N=X] represents 4 to 7 yuan of heterocyclic radicals, and X represents to constitute the needed chain of heterocyclic radical and is selected from CH, CH ₂, O, S, N or NR ₉R ₉Expression replaces or unsubstituted aryl or heterocyclic radical, and aryl refers to and replaces or unsubstituted monocycle or the dicyclic compound that contains at least one aromatic ring, and heterocyclic radical refers to and replaces or the unsubstituted heterocyclic group that contains 1 to 3 heteroatoms (N, S, P or O);

M represents the integer between the 0-6;

Described low alkyl group is the straight or branched alkyl that contains 1-6 carbon atom.

2. by the described compound of claim 1, it is characterized in that R ₅Be ethyl.

3. by the described compound of claim 2, it is characterized in that R ₁Expression C ₁-C ₃Low-grade halogenated alkyl, (CH ₂) [N=X].

4. by the described compound of claim 3, it is characterized in that described low-grade halogenated alkyl is a chloromethyl; R ₂And R ₃Perhaps R ₃And R ₄Formed 3 or 4 yuan of chains are methylene-dioxy or ethylenedioxy; Described [N=X] represents R ₉The piperazinyl that replaces, R ₉Expression aryl or heterocyclic radical.

5. by the described compound of claim 4, it is characterized in that described 3 yuan or 4 yuan of chains are ethylenedioxy, described aryl is a phenyl, described heterocyclic radical is a pyridyl.

6. by the described compound of claim 2, it is characterized in that the R of described compound ₁, R ₂, R ₃, R ₄Be respectively: Compound R ₄???R ₃???R ₂???R ₁ ?1??????H?????F?????H????CH ₂Cl ?2??????H?????Cl????H????CH ₂Cl ?3??????H?????Br????H????CH ₂Cl ?4??????H?????MeO???H????CH ₂Cl ?5??????H?????Me????H????CH ₂Cl ?6??????F?????H?????H????CH ₂Cl ?7??????Cl????H?????H????CH ₂Cl ?8??????Me????H?????H????CH ₂Cl ?9??????F?????F?????H????CH ₂Cl

7. the preparation method of high camptothecine compounds is characterized in that

Midbody compound D and the coupling of general formula E compound are obtained formula (I) structural compounds

Wherein, R ₁Expression H, low alkyl group, low-grade halogenated alkyl, aryl, rudimentary aralkyl, cycloalkyl, rudimentary hydroxyalkyl; R ₂, R ₃, R ₄Represent H, halogen, low alkyl group, OR independently ₈, nitro, perhaps R ₃And R ₄Form 3 or 4 yuan chain together, perhaps R ₂And R ₃Form 3 or 4 yuan chain together, the element of this chain is selected from CH ₂, O, N, S; R ₅And R ₈The expression low alkyl group, rudimentary hydroxyalkyl represents to have the low alkyl group of hydroxyl replacement.

8. the midbody compound D of the high camptothecine compounds of the described preparation of claim 7

R wherein ₅The expression low alkyl group.

9. the described midbody compound D of claim 8 is characterized in that R ₅The expression ethyl.

10. the application of the arbitrary compound in the claim 1～6 in preparation topoisomerase enzyme inhibitor, antitumor drug or antiviral.