CN102086207A - Camptothecin derivative with antitumor activity - Google Patents

Abstract

The invention discloses a camptothecin derivative with antitumor activity; the compound has good water solubility and highly-active antitumor effect, can be administrated orally and intravenously, and is applicable to antitumor therapy.

Description

One class has the camptothecin derivative of anti-tumor activity

Technical field

The present invention relates to anti-cancer drug compounds, more specifically, relate to the new camptothecin derivative of a class and their preparation and application with efficient antitumour activity.

Background technology

(camptothecin is the five rings alkaloid of separation and Extraction from Nyssaceae plant camptotheca acuminata (Camptotheca acuninata) CPT) to camptothecine, is the phytogenic anticarcinogen of unique selective inhibition DNA topoisomerase I (Topo I) effect.At present, camptothecine has become second important cancer therapy drug by plant derivation after taxol in oncotherapy.

Toxicity is big because camptothecine exists, weak curative effect has limited its clinical application.It is low further to seek toxicity, and the camptothecin derivative of high curative effect is the focus and emphasis of studying at present.The camptothecin derivative of early stage development has Hycamtin (Topotecan Top) and irinotecan (Irinotecan CPT-11), their toxicity is little, selectivity is good, especially irinotecan has vital role aspect clinical cancer therapy, on the treatment digestive tract tumor irreplaceable effect is arranged especially.At present just the clinical I-III phase test Gimatecan, Chimmitecan, Rubitecan, Exantecan etc. are arranged.Camptothecine is the five rings alkaloid, 7,9, and 10,11,12 and lactonic ring all can be modified, therefore, according to the female ring of camptothecine for the derivative on basis through existing hundreds of compound (CN101232872 of structural changes, 2006-08-08PCT/EP2006/065159, CN1587265, CN1616460).Most of camptothecin derivative all has certain pharmacologically active, and only effect is strong and weak different, but does not also surpass the compound of irinotecan at present in therapeutic action.

The present inventor finds that in to the screening system process of camptothecin antineoplastic agents the compound of the new texture that functional groups (structural formula (1)) such as introducing alkene or alkene derivatives obtain on 9 has good antitumor activity, thereby demonstrates the prospect of this compounds in drug development.This compounds belongs to the novel compound of invention at home and abroad also without any report.

Summary of the invention:

The present invention (1) provide a class new have an antitumour activity camptothecin derivative, (2) this compounds and their pharmaceutical compositions and (3) their anti-tumor activity.

(1) new camptothecin derivative basic chemical structure is as follows:

In the structural formula:

R1 representative: H, F, C ₁-C ₄The straight or branched alkyl

R2 representative: H, F, C ₁-C ₄The straight or branched alkyl

R3 representative: H, F, C ₁-C ₄The straight or branched alkyl

Described pharmaceutically useful salt be characterized as the amido and the medicinal mineral acid or the salt of organic acid formation that (4 '-piperidinyl piperidine) carbonyl oxygen group on 10 has alkalescence, these salt can make medicine become water-soluble.For example: hydrochloride, hydrobromate, phosphoric acid salt, vitriol, acetate, trifluoroacetate, Citrate trianion, maleate, oxalate, succinate, benzoate, tartrate, fumarate, mandelate, ascorbate salt, malate, mesylate, tosilate etc.

The present invention particularly preferably is following compound:

Compound 1:10-[(4 '-piperidinyl piperidine) carbonyl oxygen base]-9-vinyl camptothecine

Compound 2:10-[(4 '-piperidinyl piperidine) carbonyl oxygen base]-9-(1 ', 1 '-difluoro) vinyl camptothecine

Compound 3:10-[(4 '-piperidinyl piperidine) carbonyl oxygen base]-9-(1 ', 1 '-dimethyl) vinyl camptothecine

The present invention also provides the preparation method of compound of the present invention as shown below:

(2) the present invention also comprises the pharmaceutical composition of forming with the compound of effective dose of the present invention.Described pharmaceutical composition can be used for oral administration and drug administration by injection.

Oral administered dosage form can be tablet, capsule, oral liquid, granule, electuary, suspensoid etc.The used medical dressing of oral administered dosage form is conventional dressing, comprises thinner, seasonings, solubilizing agent, lubricant, suspension agent, tackiness agent, swelling agent etc.

But injection injection liquid, powder injection, lyophilized powder, liposome etc.The used medical dressing of injection is conventional dressing, comprises solvent, thinner, solubilizing agent, pH regulator agent etc.

Effective dose is meant the therapeutic action dosage that patient is produced beneficial effect, and its consumption is according to route of administration, dosage regimen and difference.

The camptothecin derivative anti-tumor activity that (3) one classes are new.

[1]. new camptothecin derivative is in external biological activity

The new camptothecin derivative of table 1 is in external cytotoxicity with to the effect of topoisomerase (Topo I)

A:IC ₅₀For suppressing the concentration of 50% cell growth.

B：+：＞20nM，++：5-20nM，+++：1-2nM++++++++：0.1-1nM，++++++++：＜0.1nM

Experimental technique: mtt assay: get one bottle in cell in good condition, add 0.25% trysinization liquid (MX-1 and OV-3 cell), digestion comes off attached cell, counting 5X10 ^-4Individual/m, make cell suspension.The CCRF cell uses suspension.

1. obtained cell suspension is inoculated on 96 orifice plates, and constant temperature CO is put in the 200ul/ hole ₂Cultivated 24 hours in the incubator.

2. change liquid, adding is subjected to the reagent thing, and cultivated 48 hours in the 20ul/ hole.

3. MTT is added in 96 orifice plates, the 20ul/ hole, reaction is 4 hours in the incubator

4. inhale and remove supernatant liquor, add DMSO, the 20ul/ hole, jolting is 5 minutes on the dull and stereotyped shaking table.

Is the light absorption value that the 570nm place measures every hole with enzyme-linked immunosorbent assay instrument at wavelength.

[2]. new camptothecin derivative is to the effect of human implantation's tumor treatment:

Experimental technique: laboratory animal is a nude mice, establishes control group and treatment group.Transplantation tumor is got eugonic tumor tissues, is cut into thin piece with sterile scissors, the tumor tissues of every inoculation 50mg.Treatment is beginning in the tenth day after transplantation tumor, and this moment, tumor weight was about 300mg.Every other day claim the nude mice body weight and measure gross tumor volume.Tried transplantation tumor: MX-1: human breast carcinoma, A549: people's nonsmall-cell lung cancer, OV-3: human ovarian cancer.The IV=intravenous injection, the PO=gastric infusion.

The new camptothecin derivative of table 2 is to the effect of human implantation's tumor treatment

A: tumour inhibiting rate (%)=(control group knurl volume-treatment group volume)/control group volume X100%. is in the back 30 days result of experiment.

B: tumour disappears: visual inspection exists less than tumour, in the back 30 days result of experiment.

C: body weight change is measuring in lower-most point in body weight.Body weight X100% before body weight change (%)=(body weight after body weight before the treatment-treatment group)/treatment.

Embodiment

Further specify the present invention by the following examples, but not as limitation of the present invention.

Embodiment 1

The concrete preparation method of compound 1:

The preparation of midbody compound A:

10-hydroxycamptothecine (365mg, 1 equivalent) is dissolved in 1, in the 4-dioxane (5.0ml). in this solution, add N-iodosuccinimide (225mg, 1 equivalent) at 0 ℃.Stirred at ambient temperature 3 hours.Final solution is spin-dried under vacuum, and remaining solid recrystallization in ethanol obtains pure product compd A 400mg (productive rate 82%).

The preparation of midbody compound B:

Compd A (400mg, 1 equivalent) is dissolved in 1, in the 4-dioxane (5.0ml), adds chloromethyl methyl ether (0.2ml, 3 equivalents) at 0 ℃ in this solution, and N, N '-diisopropylethylamine (0.4ml, 3 equivalents).Stirred at ambient temperature 10 hours.Final solution is spin-dried under vacuum, and final product obtains pure product compd B 395mg (productive rate 90%) with the silica gel column chromatography separation.

The preparation of midbody compound C:

Compd B (395mg, 1 equivalent), vinyl three potassium fluoborate salt (130mg, 1.2 equivalent), Potassium monofluoride (144mg, 3 equivalents), [1,1 '-two (diphenylphosphine) ferrocene] palladium chloride (30mg, 0.05 equivalent) is dissolved in 5ml Virahol and the 0.3ml triethylamine.Under 85 degrees centigrade of warm conditions, stirred 12 hours.Final solution is spin-dried under vacuum, and final product obtains pure product Compound C 180mg (productive rate 55%) with the silica gel column chromatography separation.

The preparation of midbody compound D:

Compound C (160mg, 1 equivalent) is dissolved in methylene dichloride (5.0ml) and the methyl alcohol (5.0ml), adds concentrated hydrochloric acid 0.1ml at 0 ℃ in this solution.Stirred at ambient temperature 10 hours.Final solution is spin-dried under vacuum, and final product obtains pure product compd B 120mg (productive rate 80%) with the silica gel column chromatography separation. ¹The preparation of HNMR compound 1:

Piperidinyl piperidine chloroformic acid acid amides (80mg, 1.25 equivalents) is dissolved in the 5ml methylene dichloride, and Compound D (110mg, 1 equivalent) is dissolved in the anhydrous pyridine (5.0ml), adds above-mentioned dichloromethane solution at 0 ℃ in this solution.Being stirred to reaction at ambient temperature finishes.Final solution is spin-dried under vacuum, and final product obtains pure product compound 1150mg (productive rate 94%) with the silica gel column chromatography separation. ¹HNMR(CDCl ₃)ppm：8.65(1H，s)，8.15(1H，d)，7.65(1H，s)，7.59(1H，d)，6.96(1H，dd)，5.86(1H，d)，5.74(1H，d)，5.67(1H，d)，5.34(1H，s)，5.32(2H，s)，4.12(2H，m)，3.76(1H，bs)，2.77(4H，bs)，1.82-1.96(2H，m)，1.60-1.90(2H，m)，1.05(3H，t)

Embodiment 2

The preparation of compound 2-3,

The preparation method is identical with embodiment 1, and the raw material that only is to use is and they substituting group corresponding compounds separately.

Embodiment 3

Contain the preparation of the tablet of compound 1

Become 1000 (50mg/ sheet medicines) through pelletizing press sheet.

Embodiment 4

Capsular preparation

Prescription compound 150 grams

Microcrystalline Cellulose 200 grams

Starch 250 grams

1000 of mixing dress glue capsules contain the 50mg/ grain.

Embodiment 5

Compound 2, the tablet of compound 3 and capsule are with embodiment 3 and embodiment 4.

Embodiment 6

Compound 1 injection liquid

Transfer pH3-3.8. with hydrochloric acid and sodium hydroxide

Filter packing, sterilization.

Embodiment 7

Compound 2, the injection liquid of compound 3 is with compound 1.

Claims (7)

1. a class has the compound of structure shown in the following structural formula (1), its isomer, and corresponding body or pharmaceutically useful salt:

Wherein

R1 representative: H, F, C ₁-C ₄The straight or branched alkyl

R2 representative: H, F, C ₁-C ₄The straight or branched alkyl

R3 representative: H, F, C ₁-C ₄The straight or branched alkyl.

2. the described compound of claim 1 is characterized in that, described compound is selected from the group that comprises following compound:

Compound 1:10-[(4 '-piperidinyl piperidine) carbonyl oxygen base]-9-vinyl camptothecine

Compound 2:10-[(4 '-piperidinyl piperidine) carbonyl oxygen base]-9-(1 ', 1 '-difluoro) vinyl camptothecine

Compound 3:10-[(4 '-piperidinyl piperidine) carbonyl oxygen base]-9-(1 ', 1 '-dimethyl) vinyl camptothecine.

3. the pharmacy acceptable salt of the compound of claim 1, can be the formed salt of this compound and mineral acid or organic acid, described salt be selected from hydrochloride, hydrobromate, phosphoric acid salt, vitriol, acetate, trifluoroacetate, Citrate trianion, maleate, oxalate, succinate, benzoate, tartrate, fumarate, mandelate, ascorbate salt, malate, mesylate, tosilate.

4. a pharmaceutical composition is characterized in that comprising described compound of the claim 1 for the treatment of effective dose and conventional pharmaceutical excipient.

5. the described pharmaceutical composition of claim 4 is characterized in that described pharmaceutical composition is prepared as oral or injection type.

6. the described compound of claim 1 or its pharmacy acceptable salt are used for the treatment of application in the medicine of tumor disease in preparation.

7. the described drug regimen of claim 4 is used for the treatment of application in the medicine of tumor disease in preparation.