CN1051077C - Process for preparing benzo[c] phenanthridinium derivatives and compounds prepared by said process - Google Patents

Abstract

The present invention relates to benzo[c]phenanthridinium derivative of the general formula A, wherein each group are defined in the specification. The compounds exhibit both antitumor activity and platelet aggregation inhibition activity. In addition, hydrogen salts of the present compounds have an enhanced stability, which is an advantage in formulating the same into pharmaceutical preparations.

Description

The preparation method of benzo [C] phenanthridine derivatives

The present invention addresses the novel method of a kind of preparation benzo [C] phenanthridine derivatives, and general formula is that this derivative of A has anti-tumor activity and the thrombocyte agglomerative suppresses also active thereby can be supposed to be used as medicine.The present invention also addresses and contains the antitumor agent that described derivative is made effective ingredient.

Nowadays, alkylating agent, nucleic acid metabolism antagonist, microbiotic, plant alkaloid and other thing of the like kind have been used as cancer patients's chemotherapeutic agents.

Know: thrombosis is by hematoblastic cohesion and condense and cause, and shifts with cerebral infarction, heart trouble, cancer DIC or the like and with canceration relation is arranged.

2,3-methylene-dioxy-5-methyl-7-hydroxyl-8-methoxyl group-benzo [C] phenanthridines muriate or iodide are Chem.Pharm.Bull., the known compound described in 33,1763 (1985).From Japanese patent application KOKAI Nos.2-243628 and 3-184916 open, learn that this compound has the thrombocyte congealing activity of inhibition and anti-tumor activity.In addition, Chem.Pharm.Bull., 33,1763 (1985) have also narrated a kind of method for preparing this compound, wherein make initiator with Berberine, go on foot through tens and handle to obtain the purpose compound.

In addition, has general formula A, M wherein and N respectively do for oneself hydrogen atom and X ^-Be Cl ^-Or OH ^-5-methyl-7-hydroxyl-8-methoxyl group-benzo [C] phenanthridines muriate or oxyhydroxide be J.Org.Chem; 53, the known compound described in the 1708-1713 (1988).

Therefore, expectation can provide a kind of simple and the practical and effective method has benzo [C] phenanthridine derivatives of general formula A with preparation.

To the production of benzo [C] phenanthridine derivatives, the present invention with general formula A

The people makes excessive quantity research, and M in the formula and N represent hydroxyl or lower alkoxy separately, and perhaps M and N represent hydrogen atom simultaneously or constitute a methylene-dioxy jointly; X ^-Expression acid group or acid acid group; And R represents a low alkyl group.

Result as these researchs, had now found that a kind of production method of novelty, in fact even can produce the purpose compound with good productive rate on a large scale have only the minority step, and, wherein used the initiator different initiator used with above-mentioned currently known methods.

Have been found that in addition specific new benzo [C] phenanthridine derivatives by the preparation of this production method has anti-tumor activity and suppresses thrombocyte agglomerative activity.According to the present invention, the stability of this compound can be improved by they being changed into its acid salt, and the acid salt of these compounds has value when its formula of medicine of preparation.

Therefore, the present invention addresses a kind of preparation and has general formula A: The method of benzo [C] phenanthridine derivatives.M in the formula and N represent hydroxyl or lower alkoxy separately, and perhaps M and N represent hydrogen atom simultaneously or constitute a methylenedioxy group jointly; X ^-Expression acid group or acid acid group; And R represents low alkyl group, is characterized as the compound that general formula is C:

Wherein M and N represent hydroxyl or lower alkoxy separately, and perhaps M and N represent hydrogen atom simultaneously or constitute methylene-dioxy jointly; Y represents halogen atom; And W represents protecting group, and Compound C in the presence of organo-tin compound ring closure reaction takes place, and the oxidation aromatization takes place then and obtains the compound that general formula is D:

Wherein M and N represent hydroxyl or lower alkoxy separately, and perhaps M and N represent hydrogen atom simultaneously or constitute methylene-dioxy jointly; And W represents protecting group, the N-alkylation that reacts and carry out described compound at the Compound D and the N-alkylating agent of this generation; And the N-alkylated compound generation remove-insurance revetment manipulation that generates thus and do acid treatment to generate the purpose product of salt form.

The present invention also provides a kind of pharmaceutical composition as antitumor agent, its effective constituent, and including general formula is new benzo [C] phenanthridine derivatives of A ': M and N represent hydroxyl or lower alkoxy separately in the formula, and perhaps M and N represent hydrogen atom simultaneously or constitute methylene-dioxy jointly; X ^-Expression acid group or acid acid group; And R represents low alkyl group, and should satisfy: when M and N represent hydrogen atom simultaneously or constitute methylene-dioxy X jointly ^-During the expression acid group, R represents the low alkyl group except that methyl.

According to the present invention, the acid salt of this compound has excellent stability, thereby preponderates when its pharmaceutical preparation of prescription.

As the example of lower alkoxy, C can be proposed ₁-C ₅Alkoxyl group is methoxyl group, oxyethyl group, positive propoxy, isopropoxy, n-butoxy, isobutoxy, tert.-butoxy and pentyloxy group and other similar group for example.Preferred C ₁-C ₃Alkoxyl group such as methoxyl group, oxyethyl group, positive propoxy and isopropoxy.

The example of low alkyl group is C ₁-C ₅Alkyl, for example methyl, ethyl, n-propyl, sec.-propyl, normal-butyl, isobutyl-, the tertiary butyl and amyl group or the like.Preferred C ₁-C ₃Alkyl such as methyl, ethyl, n-propyl and sec.-propyl.

Acid group refers to constitute the acid group of normal-salt, comprises halide-ions such as chlorion Cl ^-, bromide anion Br ^-, iodide ion I ^-, fluorion F ^-Or the like, also comprise sulfate ion SO ₄ ^-, nitrate ion NO ₃ ^-, tosic acid radical ion TsO ^-Or the like; And the acid acid group refers to constitute the acid group of acid salt, wherein contains one to two hydrogen atom, for example hydrogen sulfate ion HSO ₄ ^-, dihydrogen phosphate ions H ₂PO ₄ ^-And other similar ion.In order to improve the stability of this compound, preferred employing constitutes the acid group of acid salt, and (n refers to 1 or 2, Xa to be expressed as HnXa ^-Refer to SO ₄ ^2-Or PO ₄ ^-), comprise as hydrogen sulfate ion dihydrogen phosphate ions ex hoc genus anne ion.

General formula is that the compound of C is a new compound, can below show that mode is prepared.

Initiator is a known compound [Collection Czechoslovak, Chem.Commun., 53,3184 (1988)] 2,3-dihydroxyl-5-nitro-naphthalene.This initial compounds is converted into the nitro-naphthalene compound with formula F:

Wherein M and N represent hydroxyl or lower alkoxy separately, and perhaps M and N constitute methylenedioxy group jointly.Thereafter, formula F compound is reduced to formula B compound: Wherein M and N represent hydroxyl or lower alkoxy separately, and perhaps M and N represent hydrogen atom simultaneously or constitute methylene-dioxy jointly.5-amino naphthalenes (a kind of formula B compound) is known compound and is (Merk Index, 10,6250) that can get in the city.

The naphthylamines of formula B carries out condensation with 2-(protected hydroxyl)-3-methoxyl group-6-halogeno-benzene formaldehyde under dehydration conditions, condensation product west husband (Schiff) alkali.Optionally reduce this Schiff alkali to obtain formula C compound thereafter.

Then, will carry out more detailed explanation to aforesaid method.

1. general formula is the synthesis method of the compound of F

In the formula M and N constitute the general formula F compound of methylene-dioxy jointly can be with producing someway, in this method with 2,3-dihydroxyl-5-nitro-naphthalene is dissolved in non-aqueous polar solvent such as dimethyl formamide or other allied substances, then being heated to 60 ° to 110 ℃ with methylene halide (preferred methylene dichloride or methylene bromide) in the presence of fluoride salt (preferred fluorinated caesium or Potassium monofluoride) catalyzer reacts, obtain 2,3-methylene-dioxy-5-nitro-naphthalene (Compound I).

With regard to the general formula F compound that M in the formula and N represent hydroxyl or lower alkoxy separately, this compound can be by preparation someway, with 2,3-dihydroxyl-5-nitro-naphthalene is dissolved in non-aqueous polar solvent such as dimethyl formamide or other allied substances, then at alkaline catalysts such as Li in this method ₂CO ₃Or K ₂CO ₃Exist down, under 50 to 100 ℃ temperature, carry out 10 to 20 hours alkylated reaction with haloalkane (as methyl iodide, iodoethane, sec.-propyl bromination thing or other allied substances).When 1mol haloalkane and 2,3-two chloroforms-when the 5-nitro-naphthalene reacts, will obtain two kinds of monoalkoxy compounds.Will the production dialkyl compound, preferably should use the haloalkane of 2mol at least.If a kind of lower alkoxy substitution compound is produced in expectation, and wherein N and M can adopt someway mutually not simultaneously, and the monoalkoxy compound further reacts with haloalkane after separating with stepwise process again in this method.

2. the synthesis method that has the compound of Formula B

General formula is that the compound of B can obtain for amino by the nitro that reduction has in the compound of general formula F.For carrying out this reaction, using usually can to amino any reductive agent in order to the reduction nitro.A preferable methods is included in the ethanolic soln of the following heating reflux reaction thing of palladium/carbon catalyst existence of 5-10%.

2,3-methylene-dioxy-5-amino naphthalenes also can be used J.Org.Chem., and the currently known methods described in 53,1708 (1988) obtains, though described currently known methods has the productive rate of many steps and purpose compound low.

3. the synthesis method that has the compound of general formula C

Synthetic for this reason; general formula is that the naphthylamines compound of B and 2-(protected hydroxyl)-3-methoxyl group-6-halogeno-benzene formaldehyde (can be by as J.C.S.PerkinI; 1221 (1976) and J.Org.Chem.; 53; 1708 (1988) described currently known methodss make) react; reaction conditions will be for being heated to 80 to 110 ℃ and lasting 1 to 3 hour at the reactant in toluene or the benzene, then with a kind of like this method concentrated reaction mixture: the amino by product---water---that is produced with the aldehyde radical condensation is shifted out reaction system effectively by the component distillation with toluene or benzene.Preferably be operating as: concentrate the back surplusly mix with fresh toluene or benzene again, and then heat/concentration operation.By repeating heating/concentration operation 2-4 time, might obtain dehydrating condensation product (Schiff alkali) near quantitative yield.

Then, the two keys to dehydrating condensation product condensation position carry out reduction reaction and generate the purpose compound that general formula is C.When carrying out reduction reaction, can use any reductive agent that can reduce the two keys of CN.Preferably, this is reflected at as the sodium cyanoborohydride or the dimethyamine borane existence of reductive agent and carries out under-10 to 40 ℃ low reaction temperatures down.

Next, concrete grammar of the present invention is illustrated.

A. the synthesis method that has the compound of general formula D

General formula is that the compound of C carries out ring closure reaction (eliminating the condensation reaction of hydrogen halide) in organic solvent and in the presence of organotin hydride.Preferred organotin hydride is trialkyl (C ₁-C ₆) tin hydride such as triphenyltin hydride, tri-n-butyl tin hydride, triethyltin hydride or tin trimethyl hydride, or dialkyl (C ₁-C ₆) tin hydride such as tin diphenyl hydride or di-n-butyl tin hydride.Usually, tri-n-butyl tin hydride is best and often be used.Carry out this when reaction, with the 1-6 equivalent, preferably the normal organotin hydride of 2-3 is dissolved in certain organic solvent, preferred C ₆-C ₁₀Hydrocarbon solvent such as toluene, dimethylbenzene or benzene; and preferably certain free radical reaction initiator of blending as 2,2 '-azobis isobutyronitrile, 2,2 '-azo two (2-methylbutyronitrile), 2; 2 '-azo two (2, the 4-methyl pentane nitrile), benzoyl peroxide or other allied substances.Reaction mixture be heated to 60 to 150 ℃, preferably 80 to 130 ℃ temperature 2 minutes to 4 hours, preferably 5 minutes to 1 hour to finish ring closure reaction.After this, preferably from reaction mixture, do not isolate condensation product, in the presence of oxygenant, temperature is between the oxidation aromatization that preferably carries out closed hoop between 0 and 100 ℃ between 10 and 40 ℃ 1 to 120 minute, preferably 5 to 50 minutes to obtain the compound that general formula is D.

This reaction can be used various oxygenants, includes (for example) Manganse Dioxide, lead tetraacetate, ester acid mercury and DDQ (DDQ), is preferably activated manganese dioxide.

By above-mentioned all reactions, will constitute the skeleton of benzo [C] phenanthridines.

Usually use bromine atoms as the halogen atom among the general formula C.As for the protecting group W among general formula C and the D, the hydroxyl protecting group of any kind of can use and not have any special restriction, for example acyl group (C ₂-C ₈) group such as ethanoyl and benzoyl, and C ₃To C ₆Alkyl carbonyl and C ₃-C ₁₀Alkyl such as side chain (C ₃-C ₆) alkyl and replacement or substituted benzyl not.Preferred protecting group is a benzyl series protecting group as replacing or substituted benzyl not, or collateralization alkyl (C ₃-C ₅).

B has the synthesis method of the compound of general formula A

General formula is that benzo [C] phenanthridines of D carries out the N-alkylated reaction, removes protecting group from compound then, and carrying out acid treatment thereafter is benzo [C] phenanthridine derivatives of A with the product general formula.

When carrying out the N-alkylation, general formula is that compound and the alkylating agent of D can be dissolved in organic solvent, for example C ₆-C ₁₀Hydrocarbon solvent such as dry toluene, dry-out benzene, anhydrous dimethyl benzene or other allied substances perhaps can any solvents.These reactants can be in catalyst-free or heating in the presence of salt such as alkali metal halide or carbonate (preferably Potassium Bromide, Anhydrous potassium carbonate, anhydrous sodium carbonate and other allied substances).

Temperature of reaction is generally 50 ° to 180 ℃, preferably 100 to 150 ℃.Reaction times is generally 1-24 hour, is preferably 2 to 10 hours.

Any conventional reagent that is generally used for pyridine ring N-alkylated reaction all can be used as the alkylating agent of above-mentioned reaction.Preferred alkylating agent example comprises and is used for alkylating sulfonic acid low alkyl group (C ₁-C ₄) ester such as substituted benzenesulfonic acid lower alkyl esters (and for example: the tosic acid lower alkyl esters, and more to reactive reagent as 2,4-dinitrobenzene sulfonic acid lower alkyl esters and 2-nitrobenzene-sulfonic acid lower alkyl esters) or three halogen methylsulfonic acid lower alkyl esters.Alkylating agent is (for example) methyl tosylate, 2,4-dinitrobenzene sulfonic acid ethyl ester, 2-nitrobenzene-sulfonic acid methyl esters, 2-nitrobenzene-sulfonic acid n-propyl and trifluoromethanesulfonic acid methyl esters.

Remove protecting group and will consider kind removed protecting group.For example, with regard to benzyl series protecting group or sec.-propyl, these groups can by under acidic conditions (as in the presence of) at concentrated hydrochloric acid be heated to 60 ° to 150 ℃ and be removed, with 80 to 120 ℃ for well.Reaction times is generally 0.1 to 10 hour, with 0.5 to 3 hour for well.

After protecting group is removed from compound; carry out the acid treatment of compound; for example in some way, wherein this compound is dissolved in small amount of polar solvent such as methyl alcohol or other allied substances, mixes with the acid (for example hydrochloric acid, sulfuric acid, tosic acid) that dilute with water is crossed then.The PH of solution must be lower than 4.To every mole compound, the consumption of acid is about 1 to 3 mole usually.In addition, in reaction soln, add with the miscible organic solvent of water height such as acetone to generate the salt precipitation, in dry reaction soln, it is that the compound of A is yellow powder that the result obtains general formula.

Work as X ^-When being the acid ion (for example sulfate ion) that is at least two valencys, be necessary to reduce the sulfuric acid amount of adding.If used sulfuric acid, then will generate bisul-phate (X wherein than the quantitative 1.0-2.5 molar weight doubly that manys ^-Be HSO ₄ ^-).If use the sulfuric acid of half amount, then will generate normal vitriol (X wherein ^-Be 1/2SO ₄ ^2-).

According to the present invention, this compound has following chemical property.When with the alkaline purification general formula being benzo [C] phenanthridine derivatives of A, then compound will discharge the acid of monovalent from molecule.Therefore, think that this compound may also have the intramolecularly zwitter-ion structure of expressing with following formula E:

M and N represent hydroxyl or low alkyl group separately in the formula, and perhaps M and N represent hydrogen atom simultaneously or constitute methylene-dioxy jointly; And R represents low alkyl group, and should satisfy: when M and N represented hydrogen atom simultaneously or constitute methylene-dioxy jointly, then R represented the low alkyl group except that methyl.

Perhaps, be that the compound of E also can change into the compound that general formula is A to them with the acid treatment general formula.Be present in PH with the quaternary salt structure of general formula A representative and be lower than in the solution about 4, be higher than in the solution about 4 and be present in PH with the molecule inner salt structure of general formula E representative.Therefore, when when compound is lower than solution about 4 from PH, precipitating, can obtain formula A compound.

According to the present invention, for the stability of salt shape compound, it (is acid salt: X that acid salt also contains proton (hydrogen atom) ^-Expression acid acid group) external alkaline matter had resistibility.Thereby acid-salt is more stable than the normal-salt that does not contain proton.

As if for instance, even to having 2 of relatively good character (stability), 3-methylene-dioxy-5-methyl-7-hydroxyl-8-methoxyl group-benzo [C] phenanthridines muriate exists a problem, when it at room temperature during prolonged preservation, it will decompose gradually.This compound is a kind of powdery substance that has yellow color at once after it generates.It changes its color gradually to brown.After it made three months, this compound was pitch black brown and have and be different from the character that is observed when initial.This is that it is insoluble in water owing to generated a kind of degradation production at the compound lay up period.Therefore, think that this compound does not have the character of suitable prescription drug preparation.

2,3-methylene-dioxy-5-methyl-7-hydroxyl-8-methoxyl group-benzo [C] phenanthridines hydrosulfate ((the acid group X wherein of compd A-O) ^-Be hydrogen sulfate ion HSO ₄ ^-) be a kind of foundation compound of the present invention, and can obtain with the yellow powder form.When this compound room temperature storage one month, the phase color does not have change, thus this compound in fact in this with above-mentioned muriate (acid group X wherein ^-Be chlorion Cl ^-) difference.In addition, it reaches three months even if make the back storage, with regard to regard to yellow powder attitude compound of the present invention, does not observe color change and Decomposition.0.5 2 of μ M, 3-methylene-dioxy-5-methyl-7-hydroxyl-8-methoxyl group-benzo [C] phenanthridines muriate (acid group X wherein ^-Be chlorion Cl ^-) solution ph is 4.3.On the other hand, according to 2 of the present invention 0.5 μ M, 3-methylene-dioxy-5-methyl-7-hydroxyl-8-methoxyl group-benzo [C] phenanthridines hydrosulfate (acid group X wherein ^-Be hydrogen sulfate ion HSO ^-) PH of the aqueous solution is 3.9.Therefore, the hydrosulfate compound has lower PH, thereby has recognized that described compound has higher resistibility and thereby more stable to external alkaline matter.When this compounds has acid salt form (as hydrosulfate), its pharmaceutical activity is not damaged.Therefore, by seeing that the acid salt compound has practical use as medicine on its stability that has improved.

Below, be that the representative instance of benzo [C] phenanthridine derivatives of A is shown in table 1 with general formula.Yet should be noted that according to compound of the present invention and be not limited only to those listed in the table 1 compounds.

Table 1 compound sequence number compd A-0 2,3-(methylene-dioxy)-5-methyl-7-hydroxyl-8-first

Oxygen base-benzo [C] phenanthridines salt A-1 2,3-(methylene-dioxy)-5-ethyl-7-hydroxyl-8-first

Oxygen base-benzo [C] phenanthridines salt A-2 2,3-(methylene-dioxy)-5-n-propyl-7-hydroxyl-8-

Methoxyl group-benzo [C] phenanthridines salt A-3 2,8-dimethoxy-3,7-dihydroxyl-5-methyl-benzo

[C] phenanthridines salt A-4 2,7-dihydroxyl-3,8-dimethoxy-5-methyl-benzo

[C] phenanthridines salt A-5 2-isopropoxy-3,8-dimethoxy-5-methyl-7-hydroxyl

-benzo [C] phenanthridines salt A-6 2,3,7-trihydroxy--5-methyl-8-methoxyl group-benzo

[C] phenanthridines salt A-7 5-ethyl-7-hydroxyl-8-methoxyl group-benzo [C] phenanthridines

Salt A-8 5-methyl-7-hydroxyl-8-methoxyl group-benzo [C] phenanthridines

Salt

Representative instance with intermediate compound of general formula D is shown in table 2.

Table 2 compound sequence number Compound D-1 2,3-(methylene-dioxy)-7-benzyloxy-8-methoxyl group-benzene

And [C] phenanthridines D-2 2,3-(methylene-dioxy)-7-isopropoxy-8-methoxyl group-

Benzo [C] phenanthridines D-3 2,8-dimethoxy-3-isopropoxy-7-benzyloxy-benzo

[C] phenanthridines D-4 2-isopropoxy-3,8-dimethoxy-7-benzyloxy-benzo

[C] phenanthridines D-6 2,3-diisopropoxy-7-benzyloxy-8-methoxyl group-benzo

[C] phenanthridines D-7 7-benzyloxy-8-methoxyl group-benzo [C] phenanthridines

Representative instance with intermediate compound of general formula C is shown in table 3.

Table 3 compound sequence number Compound C-1 N-(2 '-benzyloxy-3 '-methoxyl group-6 '-bromobenzyl)-

6,7-(methylene-dioxy)-naphthalidine C-3 N-(2 '-benzyloxy-3 '-methoxyl group-6 '-bromobenzyl)-

6-methoxyl group-7-isopropoxy-naphthalidine C-4 N-(2 '-benzyloxy-3 '-methoxyl group-6 '-bromobenzyl)-

6-isopropoxy-7-methoxyl group-naphthalidine C-6 N-(2 '-benzyloxy-3 '-methoxyl group-6 '-bromobenzyl)-

6,7-diisopropoxy-naphthalidine C-7 N-(2 '-benzyloxy-3 '-methoxyl group-6 '-bromobenzyl)-

Naphthalidine

Test

The drug test example is shown now.According to benzo of the present invention [C] phenanthridine derivatives the tumour cell of the various artificial culture shown in following had growth inhibitory activity.In addition, this derivative also has the thrombocyte agglomerative activity that inhibition is caused by platelet activation factor.

1. to the growth inhibitory activity of tumour cell.

In 5% carbon dioxide gas, cultivated various tumour cells 24 hours for 37 ℃, then with test medicine effect 2-4 days.With 0.05% methylenum coeruleum make cell dyeing thereafter.By extracting pigment in the cell that has dyed look.With the optical density of pigment at the 660nm place that extracts serves as that basis determines the growth inhibition ratio of cell, and calculates 50% growth inhibitory concentration (IC ₅₀).Test-results is shown in table 4.

Table 4

IC ₅₀(μg/ml)

The compound sequence number	A-0 (X -＝HSO 4 -)	A-1 (X -＝HSO 4 -)	A-2 (X -＝HSO 4 -)	A-3 (X -＝Cl -)	A-4 (X -＝Cl -)	A-6 (X -＝OTs -)	A-7 (X -＝HSO 4 -)	A-8 (X -＝HSO 4 -)
									HeLa S3 MMI SHIN-3 N-231 Lu-130 Nakajima	0.15 0.12 0.192 0.036 0.055 n.t.	0.177 0.165 0.170 0.040 0.250 0.426	n.t. 0.273 0.414 0.085 0.108 0.777	3.0 n.t. n.t. n.t. n.t. n.t.	0.201 0.996 1.06 0.199 0.291 1.87	2.1 n.t. n.t. n.t. n.t. n.t.	0.69 n.t. n.t. n.t. n.t. n.t.	0.26 0.262 0.155 0.062 0.051 n.t.

In this table, dummy suffix notation is being represented following tumour cell: HeLa S3=carcinoma colli; The MMI=ovarian cancer; The SHIN-3=ovarian cancer; The N-231=small cell lung cancer; The Lu-130=small cell lung cancer; Nakajima=cancer of the stomach; And symbol " n.t. " representative " not test (N.T.) ".Except that HeLaS, these experiments are undertaken by while controlled trial method.

2. the thrombocyte agglomerative is suppressed active

Gather the platelet rich plasma (PRP) of Japanese albino rabbit (heavy 3-4 kilogram).Use final concentration as the platelet activity factor (PAF) of 10M as the thrombocyte inducement of condensing.In PRP, add test medicine.Adding PAF makes thrombocyte generation coagulation after cultivating certain hour.Reaction stops with EDTA.After the centrifugation, remove supernatant liquor and obtain the thrombocyte throw out, in the thrombocyte throw out, add distilled water, stay serotonin in the thrombocyte and the O-phthalic aldehyde reagent generation serotonin-O-phthalic aldehyde condensate that reacts whereby.This condenses reaches mensuration wavelength 475nm place and measures under excitation wavelength 360nm.Anti-PAF activity with following formula confirmed test medicament. In equation, " value " expression " serotonin value ".

The test-results of several compounds is shown in table 5

Table 5

Test medicine compound sequence number	Concentration (ug/ml)	Inhibiting rate (%)
			A-1(X -＝HSO 4 -)	100 50 25 12.5	96.5 74.1 49.6 25.2
A-8(X -＝HSO 4 -)	100 50 25	46.4 26.5 10.9
			Physiological saline		0.0

Clearly show the trial drug inhibition activity strong from table 5 to the thrombocyte agglomerative.

3. acute toxicity

In acute toxicity test, injection is according to compound of the present invention in the female CDF1 mouse vein of giving for 6 ages in week.At dosage is mouse group survival and do not demonstrate any fatal toxicity under the 100mg/kg.

When according to general formula of the present invention being the benzo [C] of A when phenanthridine derivatives is applied medicament, these compounds can be by the prescription medicine preparation, and these preparations can be used with various usual manners.That is to say that said preparation can non-enteron aisle ground, oral, internal rectum is used or the like.Said preparation can be forms such as injection form, Powdered, granular, sheet, suppository.If necessary or demand, when the compounding pharmaceutical composition, can use various medicament auxiliarys, that is, carrier and other auxiliary material such as stablizer, sanitas, pain killer, emulsifying agent or the like.

In pharmaceutical composition, the content with benzo [C] phenanthridines powder derivative of general formula A can be according to the form of preparation and is changed in very wide scope.Usually, the composition formula of is that the content of the derivative of A is 0.01 to 10% (weight), with 0.1 to 50% (weight) for well.Remaining is as the carrier of conventional medicine composition and other auxiliary.

Dosage with benzo [C] phenanthridine derivatives of general formula A is looked patient's situation and is changed, but the common every day about 50 of being grown up is to 500mg.

According to testing data shown in top, we can say according to general formula of the present invention to be that benzo [C] phenanthridine derivatives of A not only has anti-tumor activity to various tumour cells, suppress active but also thrombocyte condensed to have, then a kind of activity it is believed that with the activity that suppresses metastasis of cancer relation.Therefore, expectation can be effectively as the medicament for the treatment of cancer according to compound of the present invention.When being used with the form of salt, use the acid salt of compound comparatively favourable, because the acid salt of compound has improved the stability of the pharmaceutical preparation that contains this change thing according to compound of the present invention.

Now, with showing that down all embodiment describe the production of several typical benzo [C] the phenanthridines father-in-law derivative with general formula A in detail.But should be noted that scope of the present invention not only is confined to these embodiment.

Embodiment 1

2, the synthesis method of 3-(methylene-dioxy)-5-nitro-naphthalene (compound 1)

Will be as the 19.3g (0.094mol) 2 of raw material, 3-dihydroxyl-5-nitro-naphthalene joins in the 290ml anhydrous dimethyl formamide with 71.5g (0.47mol) cesium fluoride.Stir the gained mixture and make into solution.In this solution, mix 6.63ml (0.103mol) methylene dichloride and be heated to 110 ℃.Afterwards, added every one hour with the amount methylene dichloride, add altogether four times, reacting by heating mixture 2 hours is so that complete reaction then.After the cooling, the reaction mixture dilute with water is also used extracted with diethyl ether.Extraction liquid concentrates then with anhydrous sodium sulfate drying, filtration.Gained residuum re-crystallizing in ethyl acetate, result obtain primary crystal 10.7g and secondary crystal 1.82g.In addition, mother liquor concentrates and purifies with silica gel column chromatography, and chromatographic column is made elutriant with petroleum ether-ethyl acetate mixture (9: 1), obtains the crystal form material of 2.00g whereby.The total amount of the compound that generates 1 is 14.55g (yield 71%).

Compound 1: yellow powder.

¹H-NMR(200MHz)，DMSO-d ₆，ppm：

6.26(s，2H)，7.52(t，J＝8.0Hz，1H)，7.58(s，1H)，7.74(s，1H)，8.55(br d，J＝8.0Hz，2H)

Embodiment 2

2, the synthesis method of 3-methylene-dioxy-5-naphthylamines (compound 2)

4.687g (0.0216mol) compound 1 is dissolved in the 200ml ethanol.Mixing 10ml hydrazine hydrate and 1g5% palladium/carbon catalyst post-heating in the gained solution refluxed 80 minutes.After the cooling, the filtering palladium/carbon catalyst is also concentrated with filtrate.The coarse crystallization material that is generated is purified with silica gel column chromatography, makes elutriant with petroleum ether-ethyl acetate (1: 2) mixture, and the result obtains 3.566g compound 2 (yield 88%).

Compound 2: fallow powder.

¹H-NMR(200MHz)，DMSO-d ₆，ppm：

6.07 (s, 2H), 5.44 (br s, 2H), 6.54 (dd, J=7.2 and 1.5Hz, 1H), 6.92 (br d, J=8.0Hz, 1H), 7.02 (dd, J=8.0 and 7.2Hz, 1H), 7.12 (s, 1H), 7.47 (s, 1H).

Embodiment 3

N-[(2 '-benzyloxy-3 '-methoxyl group-6 '-bromobenzyl)]-6,7-methylene-dioxy-naphthalidine (Compound C-1) is synthetic

3.548g (18.95mmol) compound 2 and 6.304g (19.63mmol) 2-benzyloxy-3-methoxyl group-6-bromobenzene formaldehyde are dissolved in 100ml toluene.Gained solution is heated to 110 ℃ kept 1 hour, decompression concentrates in rotatory evaporator down then.The gained residuum mixes with the 80ml fresh toluene, heating gained mixture 1 hour, and under reduced pressure to concentrate with upper type.Gained residuum (Schiff alkali) mixes to be incorporated in the ice-water bath with 50ml toluene and cools off.Then in 45 minutes in cold soln the solution of agitation and dropping 1.31g (20.85mmol) sodium cyanoborohydride in 50ml methyl alcohol.After 1 hour, reaction mixture mixes with water, and uses chloroform extraction.The organic layer anhydrous sodium sulfate drying filters reconcentration then.The gained residue is dissolved in minimum of chloroform, mixes with the equivalent sherwood oil then so that be settled out crystal in batches, crystal is with filtering to isolate and drying, chromatography Compound C-1 as a result.Mother liquor is purified with silica gel column chromatography through concentrating, and makes elutriant with sherwood oil-chloroform-ethyl acetate mixture (16: 4: 1), and the result obtains 1.415g Compound C-1.The total amount of the Compound C that generates-1 is 7.089g (yield 76%).

Compound C-1: white powder.

¹H-NMR(200MHz)，CDCl ₃，(ppm)：

3.90 (s, 3H), 4.37 (br, 1H), 4.48 (s, 2H), 5.04 (s, 2H), 5.99 (s, 2H), 6.74 (dd, J=7.4 and 1.2Hz, 1H), 6.82 (d, J=8.8Hz, 1H), 7.05 (s, 1H), 7.07 (s, 1H), 7.11 (br d, J=8.0Hz, 1H), 7.33 (d, J=8.8Hz, 1H), 7.18-7.37 (m, 6H).IR(KBr)，(cm ^-1)：3396，3096，3064，3038，2998，2944，2906，2845，1621，1603，1573，1538(s)，1501，1464(vs)，1435，1400，1393，1371，1316，1285(s)，1271(s)，1251(s)，1218，1202，1194，1173，1144，1131，1099，1072，1040(s)，993，960，948，916，907，862，831，795，778，757，739，687。

Embodiment 4

N-1 (2 '-benzyloxy)-3 '-methoxyl group-6 '-bromobenzyl]-6-methoxyl group-7-isopropoxy-naphthalidine (Compound C-3) synthetic

463mg2-methoxyl group-3-isopropoxy-5-naphthylamines and 707mg2-benzyloxy-3-methoxyl group-6-bromobenzene formaldehyde is similar to the reaction of mode shown in the embodiment 3.Yield with 90% obtains 970mg Compound C-3.

Compound C-3: white powder.

¹H-NMR(200MHz)，CDCl ₃，(ppm)：

1.36 (d, J=6.0Hz, 6H), 3.88 (s, 3H), 3.94 (s, 3H), 4.53 (s, 2H), 4.57 (septet, J=6.0Hz, 1H), 4.58 (br, 1H), 5.03 (s, 2H), 6.75 (dd, J=7.5 and 1.0Hz, 1H), 6.80 (d, J=9.0Hz, 1H), 7.08 (s, 1H), 7.09 (s, 1H), 7.13 (br d, J=7.5Hz, 1H), 7.22 (t, J=7.5Hz, 1H), 7.24-7.31 (m, 3H), 7.32 (d, J=9.0Hz, 1H), 7.34-7.41 (m, 2H).

Embodiment 5

N-[(2 '-benzyloxy)-3 '-methoxyl group-6 '-benzyl bromide]-6-isopropoxy-7-methoxyl group-naphthalidine (Compound C-4) synthetic

463mg2-isopropoxy-7-methoxyl group-5-naphthylamines and 481mg2-benzyloxy-3-methoxyl group-6-bromobenzene formaldehyde is similar to the reaction of embodiment 3 modes, obtains 800mg Compound C-4 (yield 99%).

Compound C-4: white powder.

¹H-NMR(200MHz)，CDCl ₃，(ppm)：

1.43 (d, J=6.0Hz, 6H), 3.84 (s, 3H), 3.89 (s, 3H), 4.54 (s, 2H), 4.60 (br, 1H), 4.69 (septet, J=6.0Hz, 1H), 5.04 (s, 2H), 6.76 (dd, J=7.5 and 1.0Hz, 1H), 6.81 (d, J=9.0Hz, 1H), 6.97 (s, 1H), 7.105 (s, 1H), 7.113 (br d, J=7.5Hz, 1H), 7.22 (t, J=7.5Hz, 1h), 7.24-7.30 (m, 3H), 7.33 (d, J=9.0Hz, 1H), 7.32-7.41 (m, 2H).

Embodiment 6

N-[(2 '-benzyloxy)-3 '-methoxyl group-6 '-benzyl bromide]-6,7-diisopropoxy-naphthalidine (Compound C-6) synthetic

406mg2,3-diisopropoxy-5-naphthylamines and 707mg2-benzyloxy-3-methoxyl group-6-bromobenzene formaldehyde is similar to the reaction of embodiment 3 modes, obtains 756mg Compound C-6 (yield: 89%).

Compound C-6: white powder.

¹H-NMR(200MHz)，CDCl ₃，(ppm)：

1.32 (d, J=6.1Hz, 6H), 1.39 (d, J=6.1Hz, 6H), 3.88 (s, 3H), 4.49 (septet, J=6.1Hz, 1H), 4.51 (s, 2H), 4.55 (br, 1H), 4.61 (septet, J=6.1Hz, 1H), 5.03 (s, 2H), 6.72 (dd, J=7.5 and 1.1hz, 1H), 6.80 (d, J=8.8Hz, 1H), 7.08-7.40 (m, 10H).

IR(KBr)，(cm ^-1)：3420，2978，2936，1626，1595，1581，1522，1468(s)，1437，1383，1334，1277，1253(s)，1215，1182，1159，1139，1112，1079，1012，982，954，925，860，840，800，773，735，695。

Embodiment 7

N-[(2 '-benzyloxy)-3 '-methoxyl group-6 '-benzyl bromide]-naphthalidine (Compound C-7) synthetic

1.718g5-naphthylamines and 4.239g2-benzyloxy-3-methoxyl group-6-bromobenzene formaldehyde is similar to the reaction of embodiment 3 modes and obtains 3.71g Compound C-7 (yield 62.7%).

Compound C-7: white powder.

¹H-NMR(200MHz)，CDCl ₃，(ppm)：

3.90(s，3H)，4.52(s，2H)，4.80(br，1H)，5.06(s，2H)，6.80-6.85(m，2H)，7.22-7.47(m，10H)，7.68，7.81(m，2H)

IR(KBr)，(cm ^-1)：3999(s)，3070，2944，2856，1626，1581(s)，1531，1481，1467(s)，1432，1410，1372，1355，1336，1296，1272(s)，1227，1215，1200，1178，1144，1109，1088，1073(s)，983，959，911，865，795，783，764(s)，741，690。

Embodiment 8

2,3-methylene-dioxy-7-benzyloxy-8-methoxyl group-benzo [C] phenanthridines (Compound D-1) synthetic

1.968g (4mmol) Compound C-1 and 2.91g (10mmol) tributyl tin hydrogen are dissolved in 100ml toluene, and heating gained solution to 110 ℃.In this solution, add 0.8g 2,2 '-azobis isobutyronitrile.After 30 minutes,, mix the 2g activated manganese dioxide then, and stirred 20 minutes the reaction soln cool to room temperature.Then remove manganic compound with filtering separation, and concentrated filtrate under reduced pressure.By silicagel column, wherein use chloroform-sherwood oil mixture (3: 2) to make elutriant the gained residuum.Collect main distillate fraction and concentrate and obtain coarse crystal, to wherein mixing the 25ml ethyl acetate, and the vibration heating.Admixture 200ml hexane in this solution with the crystalline material that filtering separation generated, with hexane wash and dry, provides 1.018g Compound D-1 gradually.In addition, concentrating filter liquor is used silica gel column chromatography refinement, makes elutriant with sherwood oil-chloroform-ethyl acetate mixture (16: 4: 1), result's 0.183g Compound D-1 of getting back.The total amount of the Compound D of output-1 is 1.201g (yield 73%).

Compound D-1: yellowish white powder.

¹H-NMR(200MHz)，CDCl ₃，(ppm)：

4.07 (s, 3H), 5.32 (s, 2H), 6.13 (s, 2H), 7.26 (s, 1H), 7.34-7.46 (m, 3H), 7.58 (dd, J=8.0 and 1.6Hz, 2H), 7.61 (d, J=9.0Hz, 1H), 7.84 (d, J=9.0Hz, 1H), 8.34 (d, J=9.0Hz, 1H), 8.36 (d, J=9.0Hz, 1H), 8.69 (s, 1H), 9.75 (s, 1H).

¹³C-NMR(50MHz)，CDCl ₃，(ppm)：

50.03(q)，76.21(t)，101.62(t)，102.46(d)，104.68(d)，118.57(d)，118.69(d)，119.02(d)，120.27(s)，122.48(s)，127.40(d)，128.43(s)，128.67(d)，128.87(d)x2，128.96(d)x2，129.51(s)，130.11(s)，137.59(s)，140.37(s)，144.38(s)，147.22(d)，148.71(s)，148.89(s)，149.91(s)。

IR(KBr)，(cm ^-1)：

3032，3004，2964，2944，2910，

2875，2840，1640，1616，1595，

1578，1533，1495，1460(vs)，

1440(s)，1394，1378，1358，

1324，1284(s)，1277(s)，1251

(s)，1223，1202(s)，1169，1136，

1112，1081(s)，1040(s)，991，

968，958，949，920，904，875，

850，845，830，795，759，750，

697，686，667。

Embodiment 9

2,8-dimethoxy-3-isopropoxy-7-benzyloxy-benzo [C] phenanthridines (Compound D-3) synthetic

886mg Compound C-3 is similar to the reaction of embodiment 8 modes.Obtain 512mg Compound D-3 (yield 68%).Compound D-3: yellowish white powder.

¹H-NMR(200MHz)，CDCl ₃，(ppm)：

1.54(d，J＝6.1Hz，6h)，4.04(s，3H)，4.06(s，3H)，5.05(septet，J＝6.1Hz，1H)，5.31(s，2H)，7.28(s，1H)，7.34-7.47(m，3H)，7.55-7.61(m，2H)，7.59(d，J＝9.0H.，1H)，7.86(d，J＝9.0Hz，1H)，8.33(d，J＝9.0Hz，1H)，8.35(d，J＝9.0Hz，1H)，8.77(s，1H)，9.77(s，1H)

IR(KBr)，(cm ^-1)：

2976，2938，2838，1618，1576，

1525，1509，1479，1461，1434，

1411，1389，1372，1351，1324，

1282，1265(s)，1218，1203，

1168，1143，1115，1080，1068，

1026，981，953，929，915，880，

856，842，830，797，759，747，

698，685

Embodiment 10

2-isopropoxy-3,8-dimethoxy-7-benzyloxy-benzo [C] phenanthridines (Compound D-4) synthetic

720mg Compound C-4 is similar to the reaction of embodiment 8 modes.Obtain 320mg Compound D-4 (yield 53%).

Compound D-4: yellowish white powder.

¹H-NMR(200MHz)，CDCl ₃，(ppm)：

1.51(d，J＝6.0Hz，6H)，4.07(s，3H)，4.17(s，3H)，4.80(septet，J＝6.0Hz，1H)，5.32(s，2H)，7.32(s，1H)，7.34-7.47(m，3H)，7.55-7.65(m，2H)，7.62(d，J＝9.0Hz，1H)，7.86(d，J＝9.0Hz，1H)，8.36(d，J＝9.0Hz，1H)，8.39(d，J＝9.0Hz，1H)，8.72(s，1H)，9.79(s，1H)。

IR(KBr)，(cm ^-1)：

2976，2938，2840，1618，1578，

1527，1509，1478，1456，1435，

1409，1389，1375，1355，1325，

1267(s)，1220，1191，1177，

1163，1140，1112，1083，1067，

1021，995，973，942，911，872，

849，830，804，772，759，749，

698。

Embodiment 11

2,3-diisopropoxy-7-benzyloxy-8-methoxyl group-benzo [C] phenanthridines (Compound D-6) synthetic

The reaction of similar embodiment 8 modes takes place in 692mg Compound C-6.Obtain 379mg Compound D-6 (yield 64%).

Compound D-6: yellowish white powder.

¹H-NMR(200MHz)，CDCl ₃，(ppm)：

1.45(d，J＝6.1Hz，6H)，1.z49(d，J＝6.1Hz，6H)，4.04(s，3H)，4.69(septet，J＝6.1Hz，1H)，4.92(septet，J＝6.0Hz，1H)，5.31(s，2H)，7.34-7.46(m，3H)，7.36(s，1H)，7.55-7.62(m，2H)，7.58(d，J＝9.0Hz，1H)，7.83(d，J＝9.0Hz，1H)，8.31(d，J＝9.0Hz，1H)，8.34(d，J＝9.0Hz，1H)，8.78(s，1H)，9.77(s，1H)。

IR(KBr)，(cm ^-1)：

2976，2932，1617，1576，1527，

1507，1463，1439，1413，1385，

1373，1355，1323，1265(s)，

1216，1180，1166，1139，1113，

1086，1066，998，952，932，910，

876，856，831，800，743，697。

Embodiment 12

Synthesizing of 7-benzyloxy-8-methoxyl group-benzo [C] phenanthridines (Compound D-7)

3.71g the reaction of similar embodiment 8 modes takes place in Compound C-7.Obtain 1.387g Compound D-7 (yield 46%).

Compound D-7: yellowish white powder.

¹H-NMR(200MNz)，CDCl ₃，(ppm)：

4.08 (s, 3H), 5.34 (s, 2H), 7.34-7.47 (m, 3H), 7.55-7.61 (m, 2H), 7.64 (d, J=9.0Hz, 1H), 7.61-7.71 (m, 1H), 7.71-7.81 (m, 1H), 7.97 (dd ', J=8.0 and 1.3Hz, 1H), 8.00 (d, J=9.0Hz, 1H), 8.41 (d, J=9.0Hz, 1H), 8.48 (d, J=9.0Hz, 1H), 9.36 (dd, J=8.0 and 1.0Hz, 1H), 9.84 (s, 1H).

IR(KBr)，(cm ^-1)：

3025，2985，2925，2870，2830，

1617，1574，1525，1464，1442，

1425，1355，1332，1291(s)，1281

(s)，1261，1236，1204，1177，

1154，1138，1079(s)，1024，986，

958，934，901，868，843，834，809

(s)，816，744，721，695。

The following example is addressed producing of compound with general formula A.These compounds also can exist with the another kind of form that is expressed as general formula E.As long as therefore the physical properties of back one isomer can obtain just also providing in an embodiment.

Embodiment 13

2,3-methylene-dioxy-5-methyl-7-hydroxyl-8-methoxyl group-benzo [C] phenanthridines hydrosulfate (compd A-0) synthetic.

1.00g (2.44mmol) Compound D-1,4.55g (24.4mmol) methyl tosylate and 2.91g (24.5mmol) KBr under electromagnetic force stir to be assisted mutually between thorough mixing.Heat gained mixture to 130 ℃ then and continue 5.5 hours, dilute with water use dichloromethane extraction, with anhydrous sodium sulfate drying, filtration, and concentrated.The concentrated residuum of gained mixes under magnetic agitation with 4.55g methyl tosylate and the Powdered KBr of 2.91g again, and reheat to 130 ℃ continues 3 hours.

Then, mix 5ml acetate and 2.5ml concentrated hydrochloric acid and be heated to 100 ℃ and continue 2 hours in the reaction mixture.Thereafter, the reaction mixture dilute with water adds sodium bicarbonate in batches and is neutralized.Reaction mixture usefulness dichloromethane extraction, usefulness anhydrous sodium sulfate drying, filtration also concentrate.Reaction mixture uses methylene chloride-methanol mixture (9: 1) as elutriant by silica gel column chromatography then.Collect and concentrate the cut that contains pure compound E-0.

The concentrated product that will contain compd E-0 joins in the dilute sulfuric acid aqueous solution that contains 5ml 0.5M aqueous sulfuric acid, 25ml water and ice.Concentrate the gained mixture, mix the 5ml fresh water then so that majority of compounds A-0 is dissolved in the water.Thereafter, mix 200ml acetone gradually generating yellow mercury oxide in obtained aqueous solution, precipitation is then with filtering to isolate, vacuum-drying thereby obtain 750mg compd A-0 (yield 71.2%).

Compd E-0: intense violet color solid matter

¹H-NMR(200MHz)，DMSO-d ₆，(ppm)：

3.76(s，3H)，4.41(s，3H)，6.25(s，2H)，7.04(d，J＝8.0Hz，1H)，7.20(d，J＝8.0Hz，1H)，7.55(s，1H)，7.90(d，J＝9.0Hz，1H)，7.98(s，1H)，8.27(d，J＝9.0Hz，1H)，9.02(s，1H)。

¹³C-NMR(50MHz)，DMSO-d ₆，(ppm)：

48.65(q)，55.59(q)，100.04(d)，

102.29(t)，103.22(d)，105.52(d)，

118.54(s)，119.13(d)，119.36(s)，

120.64(s)，125.41(s)，127.04(d)，

128.30(d)，131.48(s)，131.63(s)，

147.90(s)，148.09(s)，151.06(d)，

151.21(s)，168.30(s)

IR(KBr)，(cm ^-1)：

3050，2964，2910，2834，1625(s)，

1584，1559，1538(s)，1511(s)，

1501(s)，1474(s)，1459，1417，

1372(s)，1336，1287，1256(vs)，

1245(vs)，127，1180，1145，

1122，1114，1098，1042(s)，968，

940，922，891，872，864，855，

828，804，790，764，745，710，

680，660。

Compd A-0: yellow powder (X ^-=HSO ₄ ^-)

¹H-NMR(200MHz)，D ₂O，(ppm)：

3.62(s，3H)，3.94(br s，3H)，5.95(br s，2H)，6.39(s，1H)，6.78(s，1H)，6.86(d，J＝9.0Hz，1H)，6.94(d，J＝9.0Hz，1H)，7.02(d，J＝9.0Hz，1H)，7.069d，J＝9.0Hz，1H)，8.56(s，1H)

¹³C-NMR(50MHz)，D ₂O，(ppm)：

54.93 (q), 58.96 (q), 105.51 (d), 106.16 (t), 108.33 (d), 115.55 (d) and (s), 119.54 (d), 121.05 (s), 125.69 (s), 125.81 (d), 127.67 (s), 131.13 (s), 133.47 (d), 133.90 (s), 146.94 (s), 147.97 (s), 150.79 (s), 151.54 (d) and (s)

IR(KBr)，(cm ^-1)：

3476，3040，3010，2950，1628，

1603，1585，1549，1493(s)，1478

(s)，1466，1447，1415，1373，

1353，1338，1297(s)，1279(s)，

1260(s)，1218(s)，1191(s)，

1156，1112，1096，1053，1038(s)，

967，919，865，847，830，778，764，

751，709。

Embodiment 14

2,3-methylene-dioxy-5-ethyl-7-hydroxyl-8-methoxyl group-benzo [C] phenanthridines hydrosulfate (compd A-1) synthetic

With 1.433g (3.5mmol) 2,3-(methylene-dioxy)-7-benzyloxy-8-methoxyl group-benzo [C] phenanthridines (Compound D-1) mixes with 0.906ml (3.7mmol) trifluoromethanesulfonic acid ethyl ester and 10ml dry toluene, and is heated to 100 ℃ and continues 4.5 hours., to reaction mixture in mix 10ml acetate and 5ml concentrated hydrochloric acid, and be heated to 100 ℃ and continue 1 hour thereafter.Afterwards, by being added to gradually, reaction mixture makes the reaction mixture dilution in the frozen water.Add sodium bicarbonate and the neutralization reaction mixture then in batches.After this carry out extracting operation with chloroform.With anhydrous sodium sulfate drying chloroform extraction layer, filtration and concentrating under reduced pressure.The resistates of gained is purified with silica gel chromatography, wherein uses chloroform-methanol mixture (9: 1) as eluant.Collect and concentrate main segmentation and obtain the intense violet color solid matter, that is the compd E-1 with structure of representing with general formula E.Then compd E-1 is dissolved in small amount of methanol, makes its acidifying by in gained solution, adding dilute sulfuric acid aqueous solution (every mole compound E-1 consumption be 1.0 to 2.0mol) gradually.Through this step processing, the color yellowing of solution or orange.Thereafter, this solution of concentrating under reduced pressure, vibration adds down acetone in batches in concentrated aqueous solutions makes and generates precipitation, and the filtering separation precipitation is also dry, and the result obtains the compd A-1 (X wherein of yellow powder shape ^-=HSO ₄ ^-) 1.171g (yield: 75.1%).

Compd E-1: intense violet color solid matter.

¹H-NMR(200MHz)，CDCl ₃+CD ₃OD(1∶1)，(ppm)：

1.65(t，J＝7.2Hz，3H)，3.95(s，3H)，4.83(q，J＝7.2Hz，2H)，6.19(s，2H)，7.30(s，1H)，7.30(d，J＝8.2Hz，1H)，7.44(d，J＝8.2Hz，1H)，7.56(s，1H)，7.81(d，J＝8.9Hz，1H)，8.18(d，J＝8.9Hz，1H)，9.36(s，1H)

¹³C-NMR(50MHz)，CDCl ₃+CD ₃OD(1∶1)，(ppm)：

16.65(q)，54.90(t)，56.36(q)，102.71(d)，103.14(t)，103.54(d)，106.56(d)，119.61(d)，119.77(s)，120.94(d)，121.47(s)，127.20(s)，128.10(s)，129.37(d)，131.33(s)，132.58(s)，149.29(s)，149.71(s)，150.84(d)，152.12(s)，168.45(s)。

IR(KBr)，(cm ^-1)：

2970，2918，1623(s)，1582，

1558，1534(s)，1502(s)，1475

(s)，1460(s)，1376(s)，1345，

1300，1285，1250(s)，1189，

1142，1115，1099，1036(s)，941，

865，794，761，736，707，673，

664。

Compd A=1: yellow powder (X ^-=HSO ₄ ^-)

¹H-NMR(200MHz)，D ₂O，(ppm)：

1.64(br t，J＝6.3Hz，3H)，3.63(s，3H)，4.27(br m，2H)，6.02(brs，2H)，6.45(s，1H)，6.55(br s，1H)，6.81(d，J＝9.0Hz，1H)，6.95(br，s，2H)，7.05(d，J＝9.0Hz，1H)，8.79(s，1H)。

¹³C-NMR(50MHz)，D ₂O，(ppm)：

19.32 (q), 58.78 (q), 59.43 (t), 104.40 (d), 105.92 (t), 108.03 (d), 115.24 (d), 115.63 (s), 119.17 (d), 120.05 (s), 125.49 (s) and (d), 127.39 (s), 130.66 (s), 132.96 (d), 133.33 (s), 146.54 (s), 147.55 (s), 150.25 (d), 150.59 (s), 151.46 (s).

IR(KBr)，(cm ^-1)：

3364，3040，2970，2930，2888，

1622，1602，1550，1491(s)，1474

(s)，1453，1412，1383，1348，

1334，1300，1279(s)，1260(s)，

1223(s)，1217(s)，1158，1128，

1115，1103，1080，1042(s)，942，

925，884，825，771。

Embodiment 15

2,3-methylene-dioxy-5-n-propyl-7-hydroxyl-8-methoxyl group-benzo [C] phenanthridines hydrosulfate (compd A-2) synthetic

Compound D-1 and 2-nitrobenzene-sulfonic acid n-propyl carry out the reaction of similar embodiment 14 modes and output compd E-2 is handled compd E-2 with dilute sulphuric acid then in the presence of methyl alcohol, obtain compd A-2 (X wherein whereby ^-=HSO ₄ ^-).

Compd A-2: yellow powder (X ^-=HSO ₄ ^-)

¹H-NMR(200MHz)，D ₂O，(ppm)：

0.97(br t，J＝7.1Hz，3H)，1.95(br m，2H)，3.79(s，3H)，4.22(brm，2H)，6.12(br s，2H)，6.55(brs，1H)，6.65(s，1H)，7.10(d，J＝9.0Hz，1H)，7.16(d，J＝9.0Hz，1H)，7.26(d，J＝9.0Hz，1H)，7.30(d，J＝9.0Hz，1H)，8.97(s，1H)

¹³C-NMR(50MHz)D ₂O，(ppm)

12.40(q)，27.92(t)，59.22(q)，65.30(t)，104.49(d)，106.31(t)，108.62(d)，116.05(d)，116.30(s)，120.09(d)，120.79(s)，126.27(d)，126.40(s)，128.26(s)，131.31(s)，133.53(d)，134.08(s)，147.41(s)，148.36(s)，151.10(d)，151.27(s)，152.02(s)

IR(KBr)，(cm ^-1)：

3394，3064，2982，1647，1647，

1620，1602，1581，1550，1493(s)，

1472(s)，1417，1387，1375，

1355，1302(s)，1277(s)，1257

(s)，1215(s)，1172(s)，1155，

1139，1117，1105，1082，1062，

1039(s)，1001，971，934，926，

886，855(s)，829，811，790，759，

707

Embodiment 16

2,8-dimethoxy-3,7-dihydroxyl-5-methyl-benzo [C] phenanthridines father-in-law's muriate (compd A-3) synthetic

With 227mg (0.5mmol) 2,8-dimethoxy-3-isopropoxy-7-benzyloxy-benzo [C] phenanthridines (Compound D-3) mixes with 932mg (5mmol) methyl tosylate and 35mg (0.25mmol) Anhydrous potassium carbonate, and gained mixture heating up to 140 ℃ is continued 3.5 hours.Mix 6ml acetate and 3ml concentrated hydrochloric acid in the mixture again, and be heated to 100 ℃ and continue 3 hours.Afterwards, reaction mixture is added to obtains the diluted mixture thing in the frozen water gradually, neutralize to wherein adding sodium bicarbonate more in batches.Mixture after the neutralization extracts with chloroform and small amount of methanol.With anhydrous sodium sulfate drying extraction liquid, filtration, concentrating under reduced pressure.The gained resistates is purified with silica gel chromatography, wherein makes elutriant with chloroform-methanol mixture (4: 1).Collect and concentrate main segmentation and provide the intense violet color solid matter, that is to have the compd E-3 that is expressed as the general formula E structure.Then compd E-3 is dissolved in small amount of methanol, and carries out acidifying to wherein adding diluted hydrochloric acid aqueous solution (consumption is every mole compound 1.5-3.0mol) gradually, the color of compound solution becomes yellow or orange as a result.Decompression concentrated solution desolvates and excessive spirit of salt to remove.The residuum compd A-3 of gained then, its X ^-=Cl ^-The amount of the compd A that is produced-3 is 98mg (yield 52.7%)

Compd A-3: yellow powder (X ^-=Cl ^-).

¹H-NMR(200MHz)，OD ₃OD(+DCl)，(ppm)：

4.09(s，3H)，4.119s，3H)，4.95(s，3H)，7.62(s，1H)，8.08(d，J＝9.0Hz，1H)，8.16(s，1H)，8.20(d，J＝9.0Hz，1H)，8.39(d，J＝9.0Hz，1H)，8.55(d，J＝9.0Hz，1H)，9.94(s，1H)，

IR(KBr)，(cm ^-1)：

3428，3082，1621，1591，1578，

1554，1504，1448，1415，1384，

1374，1352，1339，1289(s)，1276

(s)，1220，1193，1167，1157，

1114，1061，1025，1008，966，

921，865，854，818，781，748。

Embodiment 17

2,7-dihydroxyl-3,8-dimethoxy-5-methyl-benzo [C] phenanthridines muriate (compd A-4) synthetic

2-isopropoxy-3,8-dimethoxy-7-benzyloxy-benzo [C] phenanthridines (Compound D-4) is similar to the reaction of embodiment 16 modes and produces compd E-4 with methyl tosylate, further handle compd E-4 with dilute hydrochloric acid in the presence of methyl alcohol, the result obtains compd A-4, its X ^-=Cl ^-

Compd A-4: yellow powder (X ^-=Cl ^-).

¹H-NMR(200MHz)，OD ₃OD(+DCl)，(ppm)：

4.11(s，3H)，4.17(s，3H)，5.03(s，3H)，7.49(s，1H)，8.08(d，J＝9.1Hz，1H)，8.09(s，1H)，8.12(d，J＝9.1Hz，1H)，8.39(d，J＝9.1Hz，1H)，8.57(d，J＝9.1Hz，1H)，9.98(s，1H)。

¹³C-NMR(50MHz)，CD ₃OD(+DCl)，(ppm)：

52.74(q)，57.05(q)，57.75(q)，108.55(d)，113.59(d)，115.20(d)，117.09(s)，119.49(d)，120.14(s)，125.88(d)，126.98(s)，129.99(s)，132.139d)，133.20(s)，133.46(s)，147.62(s)，150.80(s)，151.11(s)x2，151.63(d)。

IR(KBr)，(cm ^-1)：

3384，3214，1619，1589，1576，

1556，1504，1457，1445，1421，

1414，1378，1350，1338，1301(s)，

1279(s)，1262(s)，1217，1168，

1155，1115，1058，1022，1006，

976，935，866，828，818，808，

785，747，723。

Embodiment 18

2,3,7-trihydroxy--5-methyl-8-methoxyl group-benzo [C] phenanthridines toluenesulfonate (compd A-6) synthetic

2,3-diisopropoxy-7-benzyloxy-8-methoxyl group-benzo [C] phenanthridines (Compound D-6) is similar to the reaction of embodiment 16 modes and produces compd E-6 with methyl tosylate, in the presence of methyl alcohol, further handle compd E-6 with tosic acid, the result obtains compd A-6, its X ^-=OTs ^-

Compd A-6: yellow powder (X ^-=OTs ^-).

¹H-NMR(200MHz)，CD ₃OD+D ₂O(+DCl)，(ppm)

4.00(s，3H)，4.67(s，3H)，7.17(s，1H)，7.73-7.80(m，3H)，7.92(br d，J＝9.0Hz，1H)，8.35(br d，J＝9.0Hz，1H)，9.44(s，1H)；

OTs ^- moiety(Ts＝p-Toluenesu-lufonyl)：

2.38(s，3H)，7.31(br d，J＝8.0Hz，2H)，7.67(br d，J＝8.0Hz，2H)。

IR(KBr)，(cm ^-1)：

3390，3200-3050，1622，1562，

1495，1459，1454，1445，1407，

1384，1352，1339，1295(s)，1274

(s)，1217，1189，1148，1118(s)，

1056，1033，1009，978，927，871，

815，784，744，717，681，671。

Embodiment 19

Synthesizing of 5-ethyl-7-hydroxyl-8-methoxyl group-benzo [C] phenanthridines hydrosulfate (compd A-7)

183mg (0.5mmol) 7-benzyloxy-8-oxygen methyl-benzo [C] phenanthridines (Compound D-7) is mixed mutually with 0.13ml (1.0mmol) trifluoromethanesulfonic acid ethyl ester and 0.9ml dry toluene, and heating gained mixture to 100 ℃ continues 5 hours., to mixture in mix 4ml acetate and 1ml concentrated hydrochloric acid, and be heated to 100 ℃ of maintenances 1 hour thereafter.Afterwards, reaction mixture is added in the frozen water gradually.Reaction mixture after the dilution neutralizes with add sodium bicarbonate to it in batches, uses chloroform extraction again.Chloroform extraction layer anhydrous sodium sulfate drying, filtration and concentrating under reduced pressure.The gained resistates is purified with silica gel chromatography, wherein with chloroform-methanol mixture (9: 1) as elutriant.Collection and concentrated main distillate fraction obtain the intense violet color solid matter, and that is the compd E-7 that has with the represented structure of general formula E.Then compd E-7 is dissolved in small amount of methanol, gained solution is by add dilute sulfuric acid aqueous solution (consumption is every mole compound 1.0-2.0mol) and acidifying to it in batches, and the color transition of compound solution is yellow or orange as a result.Concentrating under reduced pressure compound water solution then.Vibration adds acetone down in batches and precipitates to generate in strong solution, the precipitation filtering separation, and dry, the result obtains purpose compd A-7 (its X of yellow powder shape ^-=HSO ₄ ^-) 160mg (yield 79.7%).

Compd E-7: intense violet color solid matter.

¹H-NMR(200MHz)，DMSO-d ₆，(ppm)：

1.45(t，J＝7.1Hz，3H)，3.78(s，3H)，4.82(q，J＝7.1Hz，2H)，7.10(d，J＝8.0Hz，1H)，7.21(d，J＝8.0Hz，1H)，7.63-7.76(m，2H)，8.03(d，J＝8.8Hz，1H)，8.08-8.14(m，1H)，8.32-8.41(m，1H)，8.40(d，J＝8.8Hz，1H)，9.11(s，1H)，

¹³C-NMR(50MHz)，DMSO-d ₆，(ppm)：

15.96(q)，53.13(t)，55.23(q)，99.29(d)，117.77(d)，118.88(s)，120.57(d)，123.89(s)，124.56(d)，126.38(s)，126.54(s)，126.619d)，126.68(f)，128.08(d)，128.86(d)，130.38(s)，133.47(s)，148.66(d)，151.53(s)，169.42(s)。

IR(KBr)，(cm ^-1)：

3039，2986，2938，2832，1628(s)，1614(s)，1557(s)，1530(s)，1510(s)，1495，1474，1456，1423，1386，1377(s)，1354，1345，1335，1317，1287，1266(s)，1239(s)，1194，1155，1139，1101(s)，1075，1061，1046，977，945，923，907，871，823，798，792，774(s)，752，691，675。

Compd A-7: yellow powder (X ^-=HSO ₄-)

¹H-NMR(200MHz)，D ₂O，(PPM)：

1.72 (t, J=7.0Hz, 3H), 3.56 (s, 3H), 4.52 (br q, J=7.0Hz, 2H), 6.95 and 7.05 (AB, JAB=9.2Hz, each1H), 7.21 (br s, 2H), and 7.32-7.50 (m, 2H), 7.38 (br d, J=7.9Hz, 1H), 7.59 (br d, J=7.9Hz, 1H), 8.96 (s1H)

¹³C-NMR(50MHz)，D ₂O，(ppm)：

19.21 (q), 58.82 (q), 59.83 (t), 115.65 (d), 116.24 (s), 120.59 (d), 124.28 (s), 125.66 (d), 127.01 (s) and (d), 127.52 (s), 131.05 (d), 131.38 (s) and (d), 132.25 (d), 134.28 (d), 135.72 (s), 146.64 (s), 147.90 (s), 150.21 (d)

IR(KBr)，(cm ^-1)：

3419，3052，1614，1583，1540，

1497，1468，1446，1431，1384，

1361，1341，1302(s)，1277(s)，

1216(s)，1164，1150，1143，

1130，1086，1059，985，921，854，

821，767，695

Embodiment 20

Synthesizing of 5-methyl-7-hydroxyl-8-methoxyl group-benzo [C] phenanthridines hydrosulfate (compd A-8)

The raw material of Shi Yonging is for to press J.Org.Chem. herein, and 53, the compd E-8 of the preparation of method described in the 1708-1713 (1988).Compd E-8 is similar among the embodiment 14 and is the reaction of corresponding hydrosulfate mode for transforming compd E-1.Obtain yield and be 99.0% compd A-8.

Compd A-8: yellow powder (X ^-=HSO ₄ ^-).

¹H-NMR(200MHz)，D ₂O，(ppm)：

3.42 (s, 3H), 4.08 (s, 3H), 6.79 and 6.85 (AB, JAB=9.1Hz, each 1H), 7.03 (br s, 2H), 7.16-7.24 (m, 1H), 7.24-7.34 (m, 2H), 7.63-7.73 (m, 1H), 8.61 (s, 1H).

¹³C-NMR(50MHz)，D ₂O，(ppm)：

54.98 (q), 58.81 (q), 115.64 (d), 115.87 (s), 120.58 (d), 124.93 (s), 125.69 (d), 126.84 (s), 127.36 (s), 127.98 (d), 131.01 (d), 131.60 (d) and (s), 132.41 (d), 134.66 (d), 136.01 (s), 146.82,148.09 (s), 151.29 (d).

IR(KBr)，(cm ^-1)：

3480，3000，2950，1624，1617，

1583，1537，1488，1463，1450，

1430，1381，1359，1339，1318，

1295(s)，1278(s)，1263，1241，

1228(s)，1196，1182，1167，

1143，1112(s)，1085，1066，982，

922，914，875，853，823(s)，783，

766，751，702，692，670。

Embodiment 21

Pharmaceutical composition

With 1g2,3-methylene-dioxy-5-methyl-7-hydroxyl-8-methoxyl group-benzo [C] phenanthridines father-in-law's hydrosulfate (compd A-0) (its X=HSO), the poly-solvate (polysolvate) of 1g and 1gMacrogol400 are dissolved in the 100g distilled water for injection and constitute solution.After film filter filters, divide the threading ampoule with solution, and obtain the injection preparation that per ampoule contains 50mg compd A-0 with the usual manner freeze-drying.

Embodiment 22

Pharmaceutical composition

With 1g2,3-methylene-dioxy-5-ethyl-7-hydroxyl-8-methoxyl group-benzo [C] phenanthridines father-in-law hydrosulfate (compd A-1, its X=HSO), the poly-solvate (polysoloate) of 1g and 1gMacrogol400 are dissolved in the 100g distilled water for injection and constitute solution.After film filter filters, divide the threading ampoule with solution, and obtain the injection preparation that per ampoule contains 50mg compd A-1 with the usual manner freeze-drying.

Claims (7)

1. method for preparing the benzo that general formula is A [C] phenanthridine derivatives:

M in the formula and N represent hydroxyl or lower alkoxy separately, perhaps M and N represent hydrogen atom simultaneously or constitute methylene-dioxy jointly, and X represents acid group or acid acid group, and R represents low alkyl group, this method comprises: have the compound of general formula C, deposit at organo-tin compound Under carry out ring closure reaction, and then carrying out the oxidation aromizing, to generate general formula be the compound of D:

M and N represent hydroxyl or lower alkoxy separately in the C formula, and perhaps M and N represent hydrogen atom simultaneously or together constitute methylene-dioxy, and Y represents halogen atom and W represents protecting group; M and N represent that separately hydroxyl or lower alkoxy or M and N represent hydrogen atom simultaneously or together constitute methylene-dioxy in the D formula, and W represents protecting group; The N-alkylation that general formula D compound and N-alkylating agent react and carry out described compound; The N-alkylated compound that is generated then carries out deprotection reaction and uses acid treatment with production A compound.

2. method as claimed in claim 1, X wherein ^-Expression acid acid group.

3. method as claimed in claim 1, X wherein ^-Expression hydrogen sulfate ion HSO ₄ ^-

4. method as claimed in claim 1, M wherein and N constitute methylenedioxy group jointly.

5. method as claimed in claim 4, R wherein represents methyl, ethyl or n-propyl.

6. method as claimed in claim 1, its formula of C compound is under 60-150 ℃ the temperature, carry out in the presence of tri-n-butyltin hydride and free radical reaction initiator at the ring closure reaction in the presence of the organo-tin compound, need not isolate condensation product then and just carry out the oxidation aromatization with Manganse Dioxide from reaction mixture.

7. method as claimed in claim 1, ring closure reaction wherein every normal general formula C compound with 1-6 normal tri-n-butyltin hydride do to carry out under the organo-tin compound situation.