CN1034728A - The synthetic method of dimer alkaloid compounds - Google Patents

Abstract

The present invention relates to the synthetic of dimer alkaloid compounds, specifically, is from the indoles monomer, such as vincaleucoblastine (Catharanthine) and a kind of indoline monomer, such as: Vindoline 90 synthesizes those vincaleucoblastines (Vinca) family.The rapid method of the multistep that is disclosed comprises (1), first dimerization imido  intermediate and 1,1 of 4-dihydropyridine compound, 4-reductive reactions steps is to obtain enamine: the enamine that (2) obtain oxidation under the condition of control converts second imido  intermediate to; (3) second imido  intermediate of reduction is to form target dimeric alkaloid compounds.Whole process needn't separation of intermediates, can operate in one pot to obtain the target compound class.

Description

The synthetic method of dimer alkaloid compounds

The application is the pendent Application No. No.07/011 that submitted on February 6th, 1987, the continuation of 810 part.

The present invention relates to the synthetic of dimeric alkaloid compounds, specifically, be Vinca (Vinca) family, such as vincaleucoblastine, vincristine(VCR) and leurosidine, below usually abbreviates target compound or target dimeric alkaloid compound as.

Dimeric alkaloid of the present invention has significantly antiviral, antitumor and the leukemia performance.In the past, these alkaloids are from the Changchun seeds of flowering plants, specifically separate on a small quantity from Vinca (C.roseus).Recently, synthetic make progress (United States Patent (USP) 4,144,237 and 4,279,817 of James P.kutney, and this Application No. 07/011,810) of these compounds.Unfortunately, the method for these prior arts also obtains a spot of target compound.The invention reside in and overcome the difficulty that in prior art, on yield, is met with, shockingly improve the output of this target dimeric alkaloid compound.

The present invention relates to prepare the rapid method of multistep of dimeric alkaloid compound, specifically, it is Vinca (Vinca) family, such as vincaleucoblastine, vincristine(VCR) and leurosidine, as the Application No. No:07/011 that submits on February 6th, 1987,810 is described, and this discloses as reference herein.

Briefly, the inventive method may further comprise the steps:

(a) from indoles monomer with bridge nitrogen-atoms under the cooling in approximately-77 ° to about 40 ℃ of temperature, by oxidation bridge nitrogen, form N-oxide derivative class, and need not separate described derivative;

(b) be selected from a kind of comprising at least: in the presence of acetic anhydride, halo acetic anhydride and the Acetyl Chloride 98Min. group, handle described N-oxide derivative class, to finish Pu Langnuo Paderewski (Polon ovski) type scission reaction;

(c) product that needn't separating step (b), the product of described step (b) and indoline monomer are being selected from a kind of comprising at least: in the presence of acetic anhydride, halo acetic anhydride and the Acetyl Chloride 98Min. group, in making an appointment with-70 ℃, under inert conditions, carry out the Stereoselective coupling to form first imido intermediate to about 40 ℃ low temperature;

(d) by and 1, described first imido intermediate is reduced in 4-dihydropyridine compound reaction, forms a kind of enamine thus;

(e) under the aeration condition of control, the described enamine compound by the oxidation conversion reaction obtains in step (d) prepares second imido intermediate; And

(f) product that obtains in step (e) of reduction is to form target dimeric alkaloid compound.

All above-mentioned steps, can be operated in one pot without separation of intermediates from indoles monomer and indoline monomer to final product.

The present invention relates to the synthetic of dimeric alkaloid compounds, specifically, such as: the Vinca of vincaleucoblastine, vincristine(VCR) and leurosidine (Vinca) family, represented by the structural formula I:

In the structural formula I:

Alk=CH ₃Or (CH ₂) nCH ₃, n=1-5 in the formula;

R ₁=CH ₃Or CHO

R ₂=H or CO-alkyl

R ₃＝H

R ₄=COO-alkyl or CONR ₁₃R ₁₄, R in the formula ₁₃And R ₁₄Be selected from the group of the aryl of the alkyl, aryl or the replacement that comprise hydrogen, alkyl, replacement;

Z=-CH=CH-or-CH ₂-CH ₂-

R=II or II a

In the formula

R ₇=H or COO-alkyl;

R ₈=H, OH, O-alkyl OCO alkyl or alkyl,

R ₉=H, OH, O-alkyl OCO alkyl or alkyl

R ₁₀=H, OH, O-alkyl, OCO-alkyl

R ₁₁=H or COO-alkyl, and

R ₁₂=H or alkyl.

When in the structural formula I, alkyl=CH ₃, R ₁=CH ₃, R ₂=CO-CH ₃, R ₃=H, R ₄=COOH, Z=-CH=CH-, R=structural formula II, R ₇=COOCH ₃, R ₈=OH, R ₉=C ₂H ₅And R ₁₀During=H, this product is a vincaleucoblastine; As alkyl=CH ₃, R ₁=CH ₃, R ₂=CO-CH ₃, R ₃=H, R ₄=COOCH ₃, Z=-CH=CH-, R=structural formula II, R ₇=COOCH ₃, R ₈=C ₂H ₅, R ₉=OH and R ₁₀During=H, this product is a leurosidine; And as alkyl=CH ₃, R ₁=CHO, R ₂=CO-CH ₃, R ₃=H, R ₄=COOCH ₃, Z=-CH=CH-, R=structural formula II, R ₇=COOCH ₃, R ₈=OH, R ₉=C ₂H ₅And during R=H, this product is a vincristine(VCR).

In entire chapter, disclose all alk and alkyl and represent C ₁-C ₆Alkyl, and preferably be C ₁-C ₃Alkyl, aryl are represented single aryl such as benzyl, xylyl; All percentage ratio is weight percentage; All time cycles are minute, and all temperature are a degree centigrade ℃ metering, unless have outside the specified otherwise in addition.

The method of the target dimeric alkaloid of the present invention preparation comprises from the indoles monomer of being represented by the structural formula III and begins to form a kind of N-oxide derivative:

R represents hydrogen or COO alkyl and R in the formula ₁, R ₂, R ₃And R ₄, represent H, OH, O-alkyl, OCO-alkyl, alkyl or aryl independently, this indoles monomer is oxidized, and to form a kind of N-oxide derivative, (a) is represented as the structural formula III by this bridge nitrogen of oxidation:

And relevant analogue is represented as structural formula III (b):

R in the formula ₁, R ₂, R ₃, R ₄Identical with the structural formula III, and R ₅, R ₆, R ₇And R ₈Independently, represent H or by structural formula (CH ₂) nCH ₃The alkyl of expression, n=0-10 in the formula.Preferably this indoles monomer is because step (a) is that (the structural formula III is worked as R=COOCH to vincaleucoblastine (Catharanthine) ₃And R ₁, R ₂, R ₃And R ₄During for H)

In about-77 ℃ of extremely about-40 ℃ of scopes of temperature, by with a kind of peracid such as: a chlorine peroxybenzoic acid or to a nitro peroxybenzoic acid, in a kind of inert solvent such as reaction in methylene dichloride or other the many halogenated organic solvents (step (a)), the place is oxidized at bridge nitrogen, and the N-oxide intermediate of Xing Chenging can separate and be used for next step (step (b)) thus.The such N-oxide derivative of handling that obtains in step (a), through scission reaction (step (b)) and the imido intermediate and the coupling of a kind of indoline monomer that form, form thus such as: Vindoline 90 (vindoline) is represented as the structural formula IV:

According to the C that is included in center in the aliphatic series of occupying in the N-oxide derivative ₁₈And in the indoline monomer aromatics carbon C ₁₅C-C link, it has represented the binding of two respective compound (structural formula III a and structural formula IV).

The C of indoles N-oxide derivative ₅-C ₁₈The cracked scission reaction of key such as: carry out in the presence of the trifluoroacetic anhydride reagent, for the dipolymer that fully promotes next step thereafter at C ₁₈(indoles monomer) and C ₅The coupling of (indoline monomer), before scission reaction, can add the indoline unit to this mixture, a kind of reagent that replaces the trifluoacetic anhydride component, available trichlorine acetic anhydride, acetic anhydride, acetyl halide and tolylsulfonyl acid anhydride are used for cracking and coupling.These reagent cause C in the compound ₅-C ₁₈The Pu Langnuo Paderewski type cracking of key; In structural formula III a and III b, represent.It is similar that this temperature of reaction, time and pressure condition and common Pu Langnuo Paderewski type reaction are adopted, that is: in its original application, comprise: quaternary amine and heterocyclic amine by with corresponding N-oxide compound and acetic anhydride or acetyl halide (referring to Merch Index the 8th edition, 1968.p-R03) acylation reaction take off the alkane cracking, and the temperature of coupling step is from-70 ℃ to about 40 ℃ approximately, preferably from-70 ℃ to-30 ℃ approximately approximately; And best from-60 ℃ to-40 ℃ approximately approximately.The formation of N-oxide derivative in cleavage step and coupling step, can opening or under inert gas atmosphere shrouds, carry out, described rare gas element such as zero arbitrary other rare gas element of group in argon or the periodictable, such as: helium, neon etc. or nitrogen, because the reactions steps of back needs low temperature, the reaction times of step (a)-(c) can be from several minutes to several days.Typically, step (a) should be got about 5 minutes to several hours approximately, and step (b) should be got about 5 minutes to 1 hour, and step (c) should be got about 10 minutes to several hours approximately.

The linked reaction condition of foregoing description in the methods of the invention, represented prior art [for example at United States Patent (USP) 4,279,817, Helv.chim.Acta, 59 2858(1976) and at reaction process I (Reaction Scheme I)] a kind of important improvement, especially, coupling of the present invention (step (c)), consider the unsettled dihydrogen pyridine derivative relatively that in the monomeric coupling of N-oxide derivative and indoline, forms preparation with separate.

The reaction process I

This unsettled relatively indoles-indoline dimerization intermediate can form by stereotaxis coupling step (c), it is characterized in that being positioned at the imide salt function on the atom of Nb of indoles part.This unsettled dipolymer is designated hereinafter simply as the imido intermediate.This imido intermediate is represented by structural formula V or VI:

R in the formula ₁, R ₂, R ₃, R ₄, R ₅, R ₆, R ₇And R ₈As described earlier.

As former United States Patent (USP) 4,279,817 described these prior art methods, by and alkali metal borohydride (Na BH ₄, KBH ₄, Li BH ₄) reaction, reduce this unsettled imido intermediate to obtain some stable dimeric alkaloid, for example: by reacting with the basic metal hydroborates, the imido intermediate of reduction structural formula VI, obtain 3 ', 4 '-dehydrogenation vincaleucoblastine compound, as shown in the structural formula VII and as indicated in the reaction process II.

The reaction process II

Yet, in the present invention, this unsettled relatively imido intermediate (structural formula V or VI) does not need reduction, and in fact can be separated by various chromatographic techniques, for example: described imido intermediate can use the reaction mixture of step (c) directly to adopt at suitable chromatographic system, such as: separating on post, thin layer or the high-efficient liquid phase chromatogram HPLC, preferably is anti-phase and/or particle repulsion (size exclusion) separation method.The temperature of operation can change to about room temperature from about 4 ℃.In addition, in reaction mixture with imido intermediate volatile reagent and solvent together, can under reduced pressure (be lower than about 5 mmhg), and temperature is lower than preferably under-10 ℃ and removes.Then will this residual solid (structural formula V or VI), be dissolved in the suitable organic solvent, such as: halogenated hydrocarbon, ethers, alcohols, acetonitrile is like that or various aqueous buffer solution in.The pH of this buffered soln can from about 2 to about 10.Suitable liquid buffer solution comprises: for example: phosphoric acid salt, three (methylol) aminomethane hydrochloric acid (Tris) HCl and 2-(N-morpholinyl) ethane sulfonic acid (MES) buffered soln.Imido solution then can be by the chromatographic process purifying of foregoing description, and in addition, this imido midbody solution can be directly used in reaction thereafter.

Embodiment 1 show with vincaleucoblastine (Catharanthine) (work as R=CONCH ₃And R ₁, R ₂, R ₃And R ₄During=H, the indoles monomer of structural formula III) and Vindoline 90 (the indoline monomer of structural formula IV) reaction, the preparation imido.Embodiment 1.

Prepare imido intermediate (structural formula VI) by improved Pu Langnuo Paderewski method:

This reaction is carried out under anhydrous condition.All glasswares are 120 ℃ of fully oven dry.Methylene chloride and coupling agent trifluoacetic anhydride are before use all from P ₂O ₅In distilled.

To (Catharanthine) (structural formula III, 20 milligrams, 0.6 mmole) the solution of exsiccant methylene dichloride (2 milliliters) between adding under-20 ℃ of malleation atmosphere at argon-chlorine peroxybenzoic acid (132 milligrams, 0.8 mmole), this mixture was stirred 5 minutes, and to (Cathatanthine) N-muriate (III a, the R-COOCH that form ₃; R ₁, R ₂, R ₃And R ₄=H) add the solution of Vindoline 90 (vindoline) (IV, 270 milligrams, 0.6 mmole) at methylene dichloride (1 milliliter), and this mixture is cooled to-60 ℃.Trifluoacetic anhydride (0.2 milliliter, 1.5 mmoles) is added in the reaction mixture of this stirring, remain on-60 ℃ two hours.Then, remove under vacuum in-20 ℃ and to desolvate and excessive reagent, stay this intermediate of reddish-brown residue that contains the imido intermediate and can significantly distinguish (using Waters Radial-(Pak) stopping composition C from RPLC (HPLC) ₁₈Or the CN post, methanol-water-triethylamine is a solvent systems).It shows that imido intermediate yield in this reaction of structural formula VI surpasses 80%.

Then, this imido intermediate is with 1, and the 4-dihydropyridine reduces, and this imido intermediate converts enamine compound to, as an example, and as indicated in the reaction process III.

The reaction process III

As indicated in above-mentioned, this conversion both can occur in and separate after the imido intermediate, maybe can obtain reaction mass in step (c) by handling.The imido intermediate that is adopted is depended in the production of target compound.In other words, imido intermediate V causes the generation of the similar thing of vincaleucoblastine, and imido intermediate VI causes the generation of target compound vincaleucoblastine and leurosidine Leurosidine.Otherwise vincristine(VCR) is to produce by the oxidation of vincaleucoblastine, in particular oxidation R ₁The substituting group methyl group.For example: a kind of method as described methyl oxidation is by vincaleucoblastine Jones (Jones) reagent (CrO ₃) oxidation under low-down temperature in acetone and acetic anhydride, for example :-78 ℃ (Heterocycles Vol.9 such as J.P.Kutneyt, p-201,1978).Yet, obscure in order to reduce, below will discuss at the imido intermediate of structural formula VI.Yet notice that similarly step may be carried out with the imido intermediate of structural formula V.

When initial indoles monomer has a C ₃-C ₄(that is: Catharanthine structural formula III is worked as R=COOCH to two keys ₃And R ₁, R ₂, R ₃And R ₄During=H), the coupling imido intermediate of this generation comprises α, a β-undersaturated imido functional group, as representing by the structural formula VI.This imido intermediate can be through 1 in step (d), and 4-is reduced into the represented enamine of structural formula VIII:

Be used for reductive reagent and comprise 1,4-dihydropyridine compound (being called the NADH pattern) is represented as the structural formula IX:

R in the formula ₁, R ₂, R ₃, R ₄, R ₅And R ₆, independently, can be the aryl of alkyl, aryl and the replacement of H, alkyl, replacement.Two series of these compounds take effect easily, and Chem.Rev 82 232, and 1982; Chem.Rev.72 1 1972).First series is referred to as the Chinese ester (Hantzch esters) of dwelling, at the R of structure IX ₃And R ₄In be carboxylicesters, i.e. COOC ₂H ₅Second series is 1 of N-replacement, and 4-dihydro-nicotinamide (structural formula IX) is R therein ₁Be the alkyl of replacement or the aryl functional group of replacement, for example: benzyl, and R is CONR ₇R ₈In the formula: R ₇And R ₈, independently, can be the aryl of alkyl, aryl and the replacement of hydrogen, alkyl, replacement.

Be used in 1,4-reductive 1, the preferable another kind of of 4-dihydropyridine is to be selected from 1 of structural formula IX, 4-dihydropyridine compound class, R in the formula ₂, R ₄, R ₅And R ₆Be hydrogen, R ₁Be alkyl-aryl and R ₃For-CN or

-CO-N

CONH ₂

1, an other preferable class of 4-dihydropyridine is 1,4-dihydro-nicotinamide class, R in the formula ₂, R ₄, R ₅And R ₆Be hydrogen, R ₃Be CONH ₂And R ₁Be such as the functional group of alkyl-aryl, carboxylic acid esters, carbohydrate, carboxylic acid and metal carboxylate.

The best that is used for reduction reaction is 1,4-dihydro-nicotinamide class, R in the formula ₁Be selected from the carboxylicesters and the carboxylate salt that comprise the electron rich functional group.Extensive studies shown already these 1,4-dihydro-nicotinamide class possesses so electron rich functional group, can play coordination with band positively charged imido fully, (that is: structural formula VI) increases the regiospecific (that is: 1 of the two, the 4-reduction surpasses 1,2-reduction) and the reduction ratio of imido (structural formula VI), so cause the increase of enamine (structural formula VIII) yield.These preferable and best application 1, the object lesson of 4-dihydropyridine provides to IX-J) at table 1(structural formula IX-A.

Top reductive agent can maybe can be used in combination separately.This reduction can such as: carry out in the inert atmosphere of rare gas element of organizing separately in argon or the periodictable (helium, neon etc.) or nitrogen.In reduction step (d), can adopt all kinds of SOLVENTS.Suitable solvent for example comprises: alcohols, acetonitrile or more senior member that should series, and methyl-sulphoxide, dimethyl formamide, various ethers such as dioxan and tetrahydrofuran (THF) and hydrochloric ether are usually without liquid buffer.

In the process of reduction step (d), monitor, preferably, adopt reversed-phased high performace liquid chromatographic by reaction mixture direct analysis in suitable chromatography system.This method can be used for selecting for use temperature of reaction, time, pressure and the reactant density of optimization.Temperature of reaction can be at-60 ℃ to+60 ℃ approximately approximately, and preferably from-60 ℃ to+20 ℃ approximately approximately, and best from-60 ℃ to-20 ℃ approximately approximately.This reaction times changes according to other parameter, can be from several minutes to several days.

Following experimental example is according to the reaction process III, reacts by the reductive agent with the table I from the imido intermediate, carries out synthetic (embodiment 2-12, the process A to K) of enamine.

Embodiment 2:

The reduction of imido intermediate (structural formula VI) 1-benzyl-1,4-dihydro-nicotinamide [structural formula IX, R ₁=benzyl, R ₂, R ₄, R ₅And R ₆=H; R ₃=CONH ₂; (the operating process A of structural formula IX-A)]:

With 1-benzyl-1,4-dihydro-nicotinamide (135 milligrams, 0.63 mmole, 6 equivalents) under the malleation atmosphere (greater than 760 mmhg) of argon in add under the room temperature (20 ℃) 100 milligrams in imido father-in-law intermediate (VI) in the stirred solution of 5 milliliters of degassing acetonitriles through cycle of 5 hours, then, this reaction mixture detects (Waters Radial company, weighting agent C by anti-phase HPLS high performance liquid chromatography ₁₈Or the CN post, methyl alcohol/H ₂O/ triethylamine solvent systems) monitoring, indication VI convert to fully enamine III and 3 ', 4 '-dehydrogenation vincaleucoblastine (VII) becomes the mixture (yield 75%) of 1: 1 ratio.

Embodiment 3.

The reduction of imido intermediate (structural formula VI) 1-benzyl-1,4-dihydro-nicotinamide [structural formula IX, R ₁=benzyl, R ₂, R ₄, R ₅And R ₆=H; R ₃=CONH ₂(the operating process A of structural formula IX-A)]:

To an imido intermediate (VI .100 gram) temperature dropping or add 1-benzyl-1 in 0.5 hour in 0 ℃ solution in methyl alcohol (5 milliliters) keeps in batches, 4-dihydro-nicotinamide (56 milligrams, 0.26 mmole, 2.5 equivalents) is at the solution of methyl alcohol (2 milliliters), under the malleation atmosphere (greater than 760 mmhg) of argon through cycle of 5 hours, as monitoring at embodiment 1 usefulness high performance liquid chromatography (HPLC), the indication VI convert to fully enamine VIII and 3 ', 4 '-dehydrogenation vincaleucoblastine (VII) becomes the mixture (yield 75%) of 1: 1 ratio

Embodiment 4.

As in the foregoing description 2, describing, resulting enamine VIII and 3 ', 4 '-dehydrogenation vincaleucoblastine VII with excessive sodium borohydride (500 milligrams) in 0 ℃ of processing.Then, this mixture NH ₄OH makes and is alkalescence and uses ethyl acetate extraction (3 * 200 milliliters).The organic phase dried over mgso that merges.After removing organic solvent, the product that obtains is chromatography (is the methanol/ethyl acetate wash-out with solvent systems) on preparative thin-layer chromatography silica gel, this product show be a kind of unreacted 3 ', 4 '-dehydrogenation vincaleucoblastine (VII) and compound known 4 '-deoxidation vincaleucoblastine (X, R=COOCH ₃) and 4 '-deoxidation-4 '-Biao vincaleucoblastine (X1, R=COOCH ₃) mixture.The existence of compound finally provides clearly proof to the structure of enamine VIII.

Embodiment 5.

Repeat the process of embodiment 4, the mixture that obtains at embodiment 3 with excessive sodium borohydride processing replaces the mixture at embodiment 2.This product also show be unreacted 3 ', 4 '-dehydrogenation vincaleucoblastine (VII) and known compound 4 '-deoxidation vincaleucoblastine (X, R=COOCH ₃) and 4 '-deoxidation-4 '-Biao vincaleucoblastine (X1, R=COOCH ₃) mixture.The existence of compound finally provides clearly proof to the structure of enamine VIII.

Embodiment 6.

The reduction of imido intermediate (structural formula VI) is with 3,5-di ethoxy carbonyl-2,6-dimethyl-4-phenyl-1,4-dihydropyridine [structural formula IX, R ₁=H; R ₃And R ₅=COOCH ₂CH ₃; R ₂And R ₆=CH ₃; R ₄=phenyl; (the structural formula IX=B)-process B]:

To an imido (VI that stirs, 100 milligrams) in the solution of the acetonitrile that degass (3 milliliters), be added in 3 of ethanol (12 milliliters), 5-di ethoxy carbonyl-2,6-dimethyl-4-phenyl-1, (246 milligrams of 4-dihydropyridines, 6 equivalents), the malleation atmosphere (greater than 760 mmhg) of argon down and temperature approximately under-26 ° to about 40 ℃ with this reaction mixture refluxed 3 hours.Then by anti-phase (HPLC) efficient liquid phase chromatographic analysis (as mentioned above), showing has enamine VIII and 3 in other product ', 4 '-the dehydrogenation vincaleucoblastine becomes 1: 1 ratio to generate (yield 60%)

Embodiment 7.

The reduction of imido intermediate (structural formula VI) 1-diphenyl methyl-1,4-dihydro-nicotinamide [structural formula IX, R ₁=diphenyl methyl; R ₂, R ₄, R ₅And R ₆=H; R ₃=CONH ₂; (the structural formula IX-C)-operating process C]:

To an imido intermediate (VI that stirs, 100 milligrams) in the solution of the ethanol that degass (6 milliliters), be added in the 1-phenyl methyl-1 of methyl alcohol (6 milliliters), 4-dihydro-nicotinamide (structural formula IX-C) (76 milligrams, 2.5 equivalents), under the malleation atmosphere (greater than 760 mmhg) of argon, under 20 ℃ of temperature, added 1 normal speed with per 60 minutes this reductive agent is added dropwise to.Then, by reversed-phase HPLC efficient liquid phase chromatographic analysis (weighting agent C ₁₈Or the CN post, methanol-water-triethylamine is a solvent systems) indication in other product, have enamine (VIII) and 3 ', 4 '-the dehydrogenation vincaleucoblastine becomes 0.9: 1 ratio generation.(yield 60%).

Embodiment 8

The reduction of imido intermediate (structural formula VI) 1-benzyl-3-cyano group-1,4-dihydropyridine [structural formula IX, R ₁=benzyl; R ₂, R ₄, R ₅And R ₆=H; R ₃=CN ₃(the operating process D of structural formula IX-D)]:

To an imido intermediate (VI that stirs, 100 milligrams) in the solution of the methyl alcohol that degass (6 milliliters), be added in the 1-benzyl-3-cyano group-1 of methyl alcohol (10 milliliters), 4-dihydropyridine (structural formula IV-D) (200 milligrams, 10 equivalents) under the malleation atmosphere (greater than 760 mmhg) of argon, added 1 normal speed with per 60 minutes for 20 ℃ in temperature reductive agent is added in batches.Then, reversed-phase HPLC high performance liquid chromatography (as mentioned above) is analyzed and to be indicated in other products, have enamine (VIII) and 3 ', 4 '-dehydrogenation vincaleucoblastine (VII) becomes 1: 1 ratio generation (yield 40%).

Embodiment 9

The reduction of imido intermediate (structural formula VI) 1-benzyl-1,4-dihydro-nicotinamide base-(2 '-the formamyl pyrrolidyl)-acid amides [structural formula IX, R ₁=benzyl; R ₂, R ₄, R ₅And R ₆=H; R ₃=(2 '-the formamyl pyrrolidyl) carbonyl; (the operating process E of structural formula IX-E)]:

To an imido intermediate (VI; 100 milligrams) in the solution of the methyl alcohol that degass (6 milliliters), be added in the 1-benzyl-1 of methyl alcohol (5 milliliters); 4-dihydro-nicotinamide base-(2 '-the formamyl pyrrolidyl)-acid amides [structural formula IX-E) (163 milligrams; 5 equivalents); under the malleation atmosphere (greater than 760 mmhg) of argon; in 20 ℃ of temperature, added 1 normal speed with per 30 minutes reductive agent is added in batches.Then, RPLC (HPLC) (as mentioned above) indicates in other product, enamine (VIII) and 3 ', 4 '-the dehydrogenation vincaleucoblastine becomes 1.1: 1 ratio (yield 60%).

Embodiment 10

The reduction of imido intermediate (structural formula VI) is with 1,4-dihydro-1-(1-methoxycarbonyl isobutyl--niacinamide [structural formula IX, R ₁-1-methoxycarbonyl isobutyl-: R ₂, R ₄, R ₅And R ₆=H; R ₃=CONH ₂; (the operating process F of structural formula IX-F)]:

To one stir come ammonia intermediate (structural formula VI, 100 milligrams) in the solution of the methyl alcohol that degass (6 milliliters), be added in 1 of methyl alcohol (6 milliliters), 4-dihydro-1-(1-methoxycarbonyl isobutyl--niacinamide (structural formula IX-F) (150 milligrams, 6 equivalents), under the malleation atmosphere (greater than 760 mmhg) of argon, added 1 normal speed with per 30 minutes for 20 ℃ in temperature reductive agent is added in batches.Then, analyze (as mentioned above) by RPLC (HPLC) and indicate, in other product, have enamine (VIII) and 3 ', 4 '-the dehydrogenation vincaleucoblastine forms 2: 1 ratio generation (yield 65%).

Embodiment 11

The reduction of imido intermediate (structural formula VI) 1-(2 ', 3 ', 4 ', 6 '-tetra-acetylated-(β-D-glucopyranoside base)-1,4-dihydro-nicotinamide [structural formula IX, R ₁=(2 ', 3 ', 4 ', 6 '-ethanoyl-(β)-D-glucopyranoside base; R ₂, R ₄, R ₅And R ₆=H; R ₃=CONH ₂; (the structural formula IX-G)-operating process G]:

To an imido intermediate (VI that stirs; 100 milligrams) in the solution of degassing methyl alcohol (6 milliliters), be added in the 1-(2 ' of methyl alcohol (10 milliliters); 3 '; 4 '; 6 ' tetra-acetylated (β)-D-glucopyranoside base)-1,4-dihydro-nicotinamide (structural formula XX IX (238 milligrams, 5 equivalents); in the malleation atmosphere (greater than 760 mmhg) of argon, under 20 ℃ of temperature, added 1 normal speed with per 60 minutes reductive agent is added in batches.Then, indicate in other product by rp-hplc analysis (as mentioned above), have enamine (VIII) and 3 ', 4 '-the dehydrogenation vincaleucoblastine forms 1.5: 1 ratio and generates (yield 70%).

Embodiment 12

With 1,4-dihydro-1-(2 ' methoxycarbonyl sec.-propyl)-niacinamide [molecular formula IX, R ₁=2 '-methoxy carbonyl-sec.-propyl; R ₂, R ₄, R ₅And R ₆=H; R ₃=CONH ₂; (reduction of molecular formula IX-H) imido intermediate (molecular formula VI)-step H]:

Under the positive air pressure (more than the 760mmHg) of argon and 20 ℃ of temperature, with added that 1 normal speed in batches will be in methyl alcohol (7ml) in per 30 minutes 1,4-dihydro-1-(2 '-methoxy-carbonyl sec.-propyl)-(molecular formula is the (82mg of IX-H) to niacinamide, 3.5 (VI is in the degasification methanol solution (6ml) 100mg) equivalent) to join imido (iminium) intermediate in the stirring.Subsequently, analyze (as mentioned above) with reversed phase high efficiency liquid phase HPLC and show in other product, have enamine (VIII) and 3 ' 4 ' ,-dehydrogenation vincaleucoblastine (VII) formation ratio is 1.1: the 1(productive rate is 65%).

Embodiment 13

With 1,4-dihydro-1-(1 ', 2 '-dimethoxycarbonyl ethyl)-niacinamide [molecular formula IX, R ₁=1 ', 2 ,-dimethoxy carbonyl ethyl; R ₂, R ₄, R ₅And R ₆=H; R ₃=CONH ₂; (molecular formula IX-I)-step I) imido (Iminium) intermediate (molecular formula VI) is reduced.

Under the positive air pressure (greater than 760mmHg) of argon and 20 ℃ of temperature, to imido intermediate (VI, in the methyl alcohol that degass (6ml) solution 100mg), add 1 in the methyl alcohol (10ml), 4-dihydro-1-(1 ', 2 '-the dimethoxy carbonyl ethyl)-niacinamide (molecular formula IX-I) (148mg, 5 equivalents), reductive agent added 1 normal speed with per 30 minutes and adds in batches.Subsequently, analyze (as mentioned above) with RPLC HPLD and show, in other compound, have enamine (VIII) and 3 ', 4 '-dehydrogenation vincaleucoblastine (VII) forms, ratio is 1.1: the 1(productive rate is 70%).

Embodiment 14

With 1,4-dihydro-1-(isobutyl--1-carboxylic acid sodium)-niacinamide (formula IX, R ₁=isobutyl--1-carboxylic acid sodium, R ₂, R ₄, R ₅And R ₆=H; R ₃=CONH ₂; (the step J of molecular formula IX-J)] imido intermediate (molecular formula VI) is reduced.

Under argon positive air pressure (greater than 760mmHg) and 20 ℃ of temperature, to imido intermediate (VI, add 1 in the methyl alcohol that degass (6ml) solution 100mg), 4-dihydro-1-(isobutyl--1-carboxylic acid sodium)-the niacinamide ((130mg of molecular formula IX-J), 5 equivalents) methyl alcohol (6ml) solution added 1 normal speed with 30 minutes reductive agent is added in batches.Subsequently, the hplc rp-hplc analysis of carrying out (as mentioned above) shows, in other product, have enamine (VIII) and 3 ', 4 '-dehydrogenation vincaleucoblastine (VII) forms, ratio is 2.2: the 1(productive rate is 70% 

Embodiment 15

With 1,4-dihydro-1-(isobutyl--1-carboxylic acid sodium)-niacinamide [molecular formula IX, R ₁=isobutyl--1-carboxylic acid sodium; R ₂, R ₄, R ₅And R ₆=H; R ₃=CONH ₂; The step K of molecular formula IX-J)] inferior argon father-in-law intermediate (molecular formula VI) is reduced at low temperature.

Positive air pressure (greater than 760mmHg) at argon reaches under-20 ℃ of temperature, to inferior argon intermediate (VI, in the methyl alcohol that degass (6ml) solution 100mg), add 1,4-dihydro-1-(isobutyl--1-carboxylic acid sodium)-the niacinamide ((155mg of molecular formula IX-J), 6 equivalents) methyl alcohol (6ml) solution, the reductive agent that is added once adds.After 45 minutes, under this temperature, carry out hplc rp-hplc (as mentioned above) and show, in other product, have enamine (VIII) and 3 ', 4 '-dehydrogenation vincaleucoblastine (VII X) forms, ratio is 3.2: the 1(productive rate is 80%).

Embodiment 16

Except that be reflected at-40 ℃ carry out, all the other steps repeat embodiment 15.After 60 minutes, enamine (VIII) and 3 ', 4 '-the dehydrogenation vincaleucoblastine, ratio is 4.2: the 1(productive rate is 85%) this method of this example is called step L.

The result of embodiment 2-16 is shown in table 2 and 3.

Table 2

Reductive agent is to 1 of inferior argon VI, 4-and 1, the comparison of the reductive action of 2-.

Embodiment reduction step 1,4: 1, the 2-reduzate ¹Productive rate ²(%)

2，3 A 1∶1 75

6 B 1∶1 60

7 C 0.9∶1 60

8 D 1∶1 40

9 E 1.1∶1 60

10 F 2∶1 65

11 G 1.5∶1 70

12 H 1.1∶1 65

13 I 2.3∶1 70

14 J 2.2∶1 70

15 K 3.2∶1 80

16 L 4.2∶1 85

1. through the RPLC quantitative assay.

2.1 2-reduces (3,4-dehydrogenation vincaleucoblastine, VII) and 1, the summation of 4-reduction (enamine VIII) product.

Table 3

Temperature is to 1 of the imido VI, 4-and 1, the comparison of 2-reductive action

Embodiment reduction step temperature (℃) 1,4: 1, the 2-yied of redution ²(%)

Product ¹

14 J +20 2.2∶1 70

15 K -20 3.2∶1 80

16 L -40 4.2∶1 85

1. with high performance liquid chromatography (HPLC) quantitative assay.

2.1 2-reduces (3,4-dehydrogenation vincaleucoblastine, VII) and 1, the summation of 4-reduction (enamine VIII) product.

Table 2 shows that when the reductive agent that adopts molecular formula IX-I and molecular formula IX-J carried out reduction step (being respectively step I and J in embodiment 13 and 14), the productive rate of enamine under 20 ℃ of temperature (molecular formula VII) was the highest.Table 3 show adopt molecular formula be the reductive agent of IX-J to imido intermediate (molecular formula VI) when reducing, the temperature role.Table 3 clearly illustrates that, when reduction temperature (when reducing to-40 ℃ (step L, embodiment 16) by-20 ℃ (step K, embodiment 15), the result who is produced is, 1, the 4-reduzate is corresponding to 1,2-reduzate ratio rises, same, overall yield also increases.Even step J adopts 6 normal reductive agents and step K and L adopt 5 equivalent reductive agents, and step J(productive rate and 1,4-reductive agent product is less) the result still consistent with step K and L.

In a word, the reason of the increase of enamine (molecular formula VIII) productive rate is at imido intermediate (molecular formula VI) 1, adopts 1 in the 4-reduction, the 4-dihydropyridine, and molecular formula is IX, wherein R ₁Be an abundant electron substituents group, such as carboxylicesters and carboxylate salt.Reduction reaction is preferably under the inert atmosphere carries out, as in argon gas, and under the low temperature in about-60 ℃ to about+60 ℃ the scope, carry out, wherein be preferably about-60 ℃ extremely about+20 ℃ temperature range, and the best is the scope about-20 ℃ about-60 ℃.

The enamine (molecular formula VIII) that forms in the above-mentioned reduction step (d) can be directly used in the also available various chromatographic techniques of following reaction to be separated, for example, enamine can directly be used selected chromatographic system, a certain such as in chromatography column, thin-layer chromatography or the high performance liquid chromatography system will separate from the reaction mixture that step (d) obtains.Preferably adopt reversed phase chromatography and/or gel permeation chromatography separation method.The temperature that separating step adopts can change in the scope about room temperature to 4 ℃.On the other hand, volatile reagent and the solvent that is present in the reaction mixture can be preferably under the temperature that is lower than-10 ℃ and remove under decompression and cooling.The gained residue can be with above-mentioned chromatography method purifying in addition next carry out qualitative and before transforming.

According to currently known methods with alkali-metal boron hydrohalogenic acid salt (Na BH ₄, KBH ₄, Li BH ₄) the enamine VIII is handled, with produce 4 '-deoxidation vincaleucoblastine compound (molecular formula X, R=COOCH ₃) and 4 '-deoxidation-4 '-Biao-vincaleucoblastine compound (molecular formula XI, R=COOCH ₃).

Yet, under the oxidizing condition of the step (e) of present method, can be converted to second kind of imido intermediate representing for as molecular formula X VI and X VI a in the enamine VIII.

With enamine (molecular formula VIII) be converted into second kind of imido intermediate (molecular formula X VI and X VI a) available oxidation step comprise, for example:

(1) control ventilation/oxygenation;

(2) add the auxilliary acid of flavine (molecular formula XII):

[riboflavin, molecular formula XII, R=H; Vitamin B2 phosphate (FMN), molecular formula XII, R=PO ^2- ₃; Flavin adenine dinucleotide (FAD), molecular formula XII, R=(PO ₃) ^2- ₂-adenosine] then control ventilation/oxygenation

(3) flavine that adds reduced form is assisted acid (molecular formula X III);

[dihydroriboflavin, molecular formula X III, R=H; The dihydro flavine, mononucleotide (FMNH ₂), molecular formula X III, R=PO ^2- ₃; Dihydro flavin adenine dinucleotide (FADH ₂); Molecular formula X III, R=(PO ₃) ^2-Adenosine], then control ventilation/oxygenation

(4) adding is represented as molecular formula X III to produce accordingly as the flavocoenzyme of molecular formula XII representative, 1,5 dihydro flavocoenzyme, then control ventilation/oxygenation.

(5) add the flavocoenzyme analogue that has as the isoalloxazine structure of molecular formula X IV representative:

R wherein ₁, R ₂And R ₃, can be alkyl respectively, substituted alkyl, aryl and substituted aryl, then control ventilation/oxygenation;

(6) adding molecular formula X V is represented, the above-mentioned flavocoenzyme analogue of reduced form (1, the 5-dihydro):

R wherein ₁, R ₂And R ₃, can be alkyl respectively, substituted alkyl, aryl and substituted aryl, then control ventilation/oxygenation

(7) add hydrogen peroxide and/or as the hydroperoxide of molecular formula R-OOH, wherein R can be alkyl, substituted alkyl, aryl or substituted aryl;

(8) add as molecular formula R-CO ₃The peracid of H representative, wherein R can be alkyl, substituted alkyl, aryl or substituted aryl;

(9) add superoxide;

(10) introduce, the hydroxyl of Chan Shenging for example, adds hydrogen peroxide in the presence of ferrous ion in many ways; Or

(11) add the metal ion that can be good electron acceptor, for example, iron ion (Fe ^{+ 3}); Cuprous ion (Cu ^{+ 1}), mercurous ion (Hg ^{+ 2} ₂) and silver ions (Ag ^{+ 1}), then control ventilation/oxygenation.

Above-mentioned oxidation step (1) and (5)-(10) can be carried out in organic solvent, such as: molecular weight the higher person in alcohols, acetonitrile or this series compound; Dimethyl sulfoxide (DMSO); Dimethyl formamide; Various ethers are as diox, tetrahydrofuran (THF); And aromatic hydrocarbon, as benzene, toluene etc.

Oxidation step (condition (2), (3) and (4)) a kind of pH of needs that relates to flavocoenzyme is 5-9, be preferably the aqueous solution buffer reagent of pH6-8, for example, phosphoric acid salt, Tutofusin tris hydrochloric acid (Tris HCl) 2(N morpholine)-ethane sulfonic acid (MES) buffer reagent is as solvent.Can adopt organically and be total to broad dose, for example available alcohols; Molecular weight the higher person in acetonitrile or this series compound; Diox; Tetrahydrofuran (THF); Methyl-sulphoxide; Or dimethyl formamide.

Oxidation step carry out available a kind of suitable layers analysis system in the process, be preferably RPLC and reaction mixture is carried out direct analysis monitor.This method can be used for preferable reaction temperature, time, pressure and reactant concn.Temperature of reaction can be from about-60 ℃ to about+60 ℃ changes, and preferably adopt from about 4 ℃ the scope to room temperature.Reaction times can not waited to several days from several minutes, depends on, for example, temperature of reaction and special separately oxidizing condition.Reaction is carried out under barometric point usually.

In a conversion embodiment, from 1 of imido intermediate (molecular formula VI), 4-reduction enamine solution to obtain, under lower temperature (0 ℃ to-70 ℃) begin to use with react in 5-50 times of used same solvent dilution.The enamine solution of dilution carries out oxidation by a certain in the oxidation step described herein subsequently, and (step (e) becomes second kind of imido intermediate, and (molecular formula X VI and X VI are a).

The preferred oxidation step that present method adopts is:

(a) control ventilation/oxygenation, promptly under the situation that open enamine solution is stirred, bubbling air/oxygen flow in solution;

(b), but add iron trichloride as step (a);

(c) as step (a), but add as the flavocoenzyme of molecular formula XII representative, produce accordingly 1 immediately, 5-dihydro flavocoenzyme is represented as molecular formula X III, perhaps

(d) as step (a), but add hydrogen peroxide and/or as molecular formula R-OOH represent hydroperoxide/wherein R is an alkyl or aryl.

Oxidation step carries out in organic solvent, such as alcohols; Molecular weight the higher person in acetonitrile or the homologue; Dimethyl sulfoxide (DMSO); Dimethyl formamide; Ethers is as diox; Tetrahydrofuran (THF); Aromatic hydrocarbon is as benzene, methyl or the like.A kind of pH is 5-9, and the suitableeest is aqueous solution of buffer agent (for example, phosphoric acid salt, Tutofusin tris hydrochloric acid (Tris-HCl), 2(N-morpholinyl)-ethane sulfonic acid (MES) buffer reagent of the scope of 6-8) can be used as broad altogether dose.Temperature of reaction can be from about-60 ℃ changes to about+60 ℃.

The parameter that multiple oxidation conversion step (e) is arranged can be used for research to optimize the productive rate of required compound, particularly vincaleucoblastine (molecular formula I).The results are shown among the following table 4-7 of this class research.

Table 4

The effect of iron trichloride in enamine VIII production vincaleucoblastine

Fe Cl ₃The productive rate (%) of quantity oxidizing condition vincaleucoblastine

(equivalent) (temperature, time)

0 Air，0℃，5min 0

1 Air，0℃，5min 13.3

2 Air，0℃，5min 19.0

3 Air，0℃，5min 10.4

1. speed is 60ml/min.

2. after Na BH reduction, with the quantitative assay of reversed-phase HPLC high performance liquid chromatography.

3. the enamine VIII results from-40 ℃ (step L)

Table 5

The influence of time in enamine VIII production vincaleucoblastine

Time (minute) productive rate (%) of vincaleucoblastine

1 8.2

5 15.4

10 15.5

15 15.7

45 6.5

1. reaction conditions: add 2 equivalent iron trichlorides, the speed with 60ml/min under 0 ℃ is ventilated to solution.

2. use Na BH ₄After the reduction, use the reversed-phase HPLC quantitative assay.

3. the enamine VIII results from-40 ℃ (step L)

Table 6

From the enamine VIII ³When producing vincaleucoblastine, the effect of oxidizing temperature

Temperature ℃ ¹Vincaleucoblastine ²Productive rate (%)

-40 3.7

-23 6.2

0 19.6

20 20.6

45 16.0

1. reaction conditions: add 2 normal iron trichlorides, with the speed of 60ml/mim to solution ventilation 15 minutes.

2. with after the NaBH reduction, with the quantitative assay of reversed-phase HPLC high performance liquid chromatography.

3. enamine results from-40 ℃ (step L)

Table 7

From the enamine VIII ³When producing vincaleucoblastine, the effect of dilution

Extension rate ^1,4Vincaleucoblastine ²Productive rate

1 19.6

5 25.2

10 30.1

20 29.6

50 24.7

1. reaction conditions: add 2 normal iron trichlorides; Ventilated 15 minutes to solution 0 ℃ of speed with 60ml/min.

2. through NaBH ₄After the reduction, with the quantitative assay of reversed-phase HPLC high performance liquid phase.

3. enamine results from-40 ℃ (step L).

4. extension rate: the reductive agent (6 equivalent) that adds the molecular formula IX-J representative in the 6ml methyl alcohol in the 1-100mg imido VI in 6ml methyl alcohol.(cumulative volume=12ml)

Extension rate 5: cumulative volume is 6ml;

Extension rate 10: cumulative volume is 120ml;

Extension rate 20: cumulative volume is 240ml;

Extension rate 50: cumulative volume is 600ml;

Table 4 shows the influence of ferric chloride concn to vincaleucoblastine (I) productive rate, shows the rising along with ferric chloride concn, and productive rate rose before this, but descends subsequently.The concentration that produces the iron trichloride of the highest vincaleucoblastine is about 2 equivalents.

In the table 5, enumerate productive rate, show when having 2 equivalent iron trichlorides corresponding to the vincaleucoblastine (I) of different oxidization times, aeration time is to about 15 minutes about 5 minutes, vincaleucoblastine obtains maximum productive rate, and when oxidization time was 45 minutes, productive rate obviously descended.

In the table 6, enumerate of the influence of various oxidizing temperature, show when using NaBH to the productive rate of vincaleucoblastine (I) ₄The RPLC quantitative assay of reduction back, definite when temperature from about 0 ℃ during the scope to about 20 ℃, can obtain the maximum output of vincaleucoblastine.

In the table 7, enumerate of the influence of the various extent of dilution of enamine (VIII) solution, show in the presence of iron trichloride (2 equivalent), under 0 ℃ of temperature vincaleucoblastine production, when the extension rate of enamine (VIII) solution before the ventilation is about 5 to 20 left and right sides, using NaBH ₄After the reduction, determine to obtain the maximum output of vincaleucoblastine (I) through the RPLC quantitative assay.

The listed result of table of induction 4-7 as can be known, in the oxidation switching process (step (e)) of enamine, by imines father-in-law intermediate (molecular formula VI) through 1, the extension rate of used same solvent dilution when 4-reduction and the solution of the enamine that obtains are reduced, to be positioned at 5 to 20 times scope is good, wherein is preferably (8 to 12 times) about 10 times.Dilution step carry out at low temperatures (from about 0 ℃ to-70 ℃) be preferably and be lower than about-40 ℃, and at inert atmosphere, as under argon gas shrouds, carrying out.The oxidation conversion process of enamine is preferably undertaken by following data: (1) was preferable with 60ml/min speed ventilation 5 to 20 minutes with about 15 minutes; (2) carrying out to 20 ℃ of left and right sides temperature ranges about 0 ℃, is preferable with 20 ℃; (3) in the presence of 2 equivalent iron trichlorides, (molecular formula X VI and VI are a) to obtain corresponding second kind of imines father-in-law intermediate.

Step (f) by with a kind of alkali metal borohydride (Na BH ₄, KBH ₄, LiBH ₄Or the like) (molecular formula X VI and X VI a) obtain target compound vincaleucoblastine and leurosidine (Leurosidine) to reduce these second kind of imido intermediate, and by product 3 ', 4 '-dehydrogenation vincaleucoblastine (molecular formula VII), Bai Nuosheng (Leurosine) (molecular formula X VII), (catharine) (molecular formula X VIII), (vinamidine) (molecular formula X IX) and reduzate (molecular formula XX) (Vinamidine).

Reduction process is carried out in suitable solvent (organic or inorganic), and described solvent for example is the above-mentioned solvent that is used for oxidation conversion step (e).Reduction process is in lower temperature, carries out in about about-20 ℃ to 4 ℃ scope, is preferably about 0 ℃, and carries out in the pH value that is lower than 8.5, is preferably in about 7.5 to about 8 scope.Total mixed solution of reaction can be in the lower temperature range about to 10 ℃ from about 0 ℃ in extracting or before separating target compound, and vacuum and low temperature concentrates.

Following embodiment (embodiment 17-21, method 1-5) is for describing the method for oxidation enamine (molecular formula VIII) with synthetic vincaleucoblastine.

Embodiment 17

With vitamin B2 phosphate (FMN molecular formula XII, R=PO ^-2 ₃) enamine (molecular formula VIII) is oxidized into imine intermediate (X VI) and synthetic vincaleucoblastine-method 1.

(embodiment 2 by method as mentioned above by intermediate (VI) to containing, step (A) obtains being dissolved in three (methylol) aminomethane hydrochloric acid (FMN(80mg of damping fluid (2ml), 1 equivalent) adding of enamine (VIII) 100mg in the reaction mixture of stirring under the positive air pressure (greater than 760mmHg) of argon.This solution kept in Dark Place 16 hours under room temperature (20 ℃).Afterwards, replace the inert argon atmospher with air, and with reaction mixture restir 2.5 hours.Rp-hplc analysis shows that enamine (VIII) transforms into imido father-in-law intermediate X VI and other by product.Add sodium borohydride (500mg) down at 0 ℃, use NH ₄OH makes reaction mixture be alkalescence and (3 * 200ml) extract with ethyl acetate.The organic extract that merges with dried over mgso after, removal of solvent under reduced pressure obtains crude product (85mg).Thick-layer chromatography (silica gel, the methyl alcohol: ethyl acetate 1: 5) can separate the dipolymer product of following composition: vincaleucoblastine (molecular formula I, 22mg, 23%) of this crude product are used to purify; 3 ', 4 '-dehydrogenation vincaleucoblastine (molecular formula VII, 16mg, 17%); Bai Nuosheng (Leurosine) (molecular formula X VI, 8mg, 9%), (catharine) (molecular formula X VIII, 7mg, 7.5%); (Vinamidine) (molecular formula X IX, 5mg, 5.6%) and reduzate (molecular formula XX, 19mg, 20%) (Vinamidine).

Embodiment 18

With hydrogen peroxide enamine (molecular formula VIII) is oxidized to imido intermediate (molecular formula X VI) and is used for synthesizing vincaleucoblastine (molecular formula I)-method 2.

In the solution that contains enamine (VIII) 100mg that obtains by imido intermediate VI (embodiment 2, steps A), under the inert argon atmospher, add hydrogen peroxide (30%, 1.2ml, 95 equivalents).Reaction mixture was stirred under room temperature 5.5 hours.At that time, show that through rp-hplc analysis the enamine VIII transforms fully, add sodium borohydride (500mg) down at 0 ℃, and (3 * 200ml) extract the gained solution with ethyl acetate.The organic extract liquid that merges removes with dried over mgso and under vacuum and desolvates.The mix products that is obtained gets following alkaloid constituent with thick-layer chromatography (silica gel, methanol/ethyl acetate) separation: vincaleucoblastine (I, 4mg, 4%), 3 ', 4 '-dehydrogenation vincaleucoblastine (VII, 5mg, 4.8%), Bai Nuosheng (Leurosine) (X VII, 13mg, 12.5%), (catharine) (X VIII, 5mg, 4.8%), (Vinamidine) (XX of reduced form, 30mg, 27.6%).

Embodiment 19

Become imido intermediate (molecular formula X VI) to be used for synthesizing vincaleucoblastine (I)-method 3 with atmospheric oxidation enamine (VIII).

The solution that will contain enamine (VIII) 100mg that is obtained (embodiment 2, steps A) by imido intermediate VI is under the situation that is open to air, in stirring at room 3 hours.After this, add sodium borohydride (500mg) down, use NH at 0 ℃ ₄OH makes reaction mixture be alkalescence, and (3 * 200ml) extract with ethyl acetate.The organic extract liquid MgSO that merges ₄Drying, solvent is removed under vacuum.The crude product that obtains separates with thick-layer chromatography (silica gel, methanol/ethyl acetate), obtains vincaleucoblastine (I, 4mg, 4%).

Embodiment 20

In the presence of iron trichloride, become imido father-in-law intermediate (X VI) and be used for synthesizing vincaleucoblastine (I)-method 4 with atmospheric oxidation enamine (molecular formula VIII).

In the solution that contains enamine (VIII) 100mg that obtains by imido intermediate VI (embodiment 2, steps A) through stirring, under 0 ℃, add iron trichloride (1 equivalent), and in solution blowing air 0.5 hour.Add sodium borohydride (500mg) down at 0 ℃, (before 3 * 100ml) extractions, reaction mixture is used NH earlier using ethyl acetate ₄OH furnishing alkalescence.Use MgSO ₄The dry extraction liquid that merges, solvent removed in vacuo.With thick-layer chromatography (silica gel, methanol/ethyl acetate) purification crude product obtain vincaleucoblastine (I, 37mg).Be as the criterion the productive rate 70% of vincaleucoblastine with the enamine (50mg) that exists in the mixed solution.

Embodiment 21

Under high extent of dilution, under the iron trichloride existence condition, synthesize vincaleucoblastine (molecular formula) I-method 5 to imido intermediate (molecular formula X VI) with atmospheric oxidation enamine (molecular formula VIII).

To contain the solution that obtains enamine (VIII) 200mg by imido intermediate VI (embodiment 16, step L), before oxidation, dilute 5 times of (cumulative volumes: 120ml) with methyl alcohol.Then, add iron trichloride (75mg, 2 equivalents), under 0 ℃, to solvent blowing air 20 minutes.Subsequently, add sodium borohydride (200mg),, add entry (100ml) subsequently, and (3 * 200ml) extract with ethyl acetate solution for vacuum concentration.The organic extract liquid Na that merges ₂SO ₄Drying, vacuum is removed solvent.With column chromatography (silica gel, TLC(thin-layer chromatography) level, 15g) purification crude product.Use ether: chloroform (10: 7) wash-out gets 3 ', 4 '-dehydrogenation vincaleucoblastine (VII, 18mg, 11%).Use ether instead: chloroform: methyl alcohol (10: 7: 0.5) wash-out obtains vincaleucoblastine (I, 62mg, 37%).

For actual purpose, must (V, VI, VIII, X VI, X VI a) not be separated to intermediate, best for all processes of indoles unit (molecular formula III) and indoline unit (molecular formula IV) as embodiment 22 described modes, carry out one pot of (one-pot) operation.

Embodiment 22

Vincaleucoblastine (Catharanthine) (molecular formula III, R ₁, R ₂, R ₃And R ₄=H, R=COOCH ₃) and vindoline (molecular formula III) become the whole process of one pot of conversion of vincaleucoblastine (molecular formula I) and leurosidine (Leurosidine).

Under-15 ℃, the positive air pressure of argon (greater than＞760mmHg) under, between inciting somebody to action-chlorine peroxybenzoic acid (330mg, 1.9mmol) once add vincaleucoblastine (catharanthine) (500mg, 1.5mmol) dry dichloromethane (4.5ml) solution in, under 10 ℃ to-15 ℃, this mixed solution was stirred 5 minutes.Subsequently, reaction solution is cooled to-40 ℃, add exsiccant methylene dichloride (1ml) Vindoline 90 (IV, 450mg, 1mmol) solution, and then add trifluoroacetic anhydride (1ml, 7.1mmol).After-60 ℃ were placed 2 hours, vacuum was removed volatile matter (high-vacuum pump), and dry, after this system is by argon cleaning, added the methyl alcohol (12ml) that degass.Orange solution with obtaining is cooled to-40 ℃, and adds 1 under the positive air pressure of argon, 4-dihydro-1-(isobutyl--1-carboxylic acid sodium)-niacinamide (molecular formula IX-J) (1.5g, 6mmol) solution in the methyl alcohol of doing that degass (12ml).When reduction fully back (with reversed phase high efficiency liquid phase HPCL monitoring), adding cold methanol (about 300ml) remains on the temperature of this solution about-5 ℃ to about 0 ℃ scope.Subsequently, (330mg 2mmol), and fed dry air 20 minutes with the speed about 60ml/min to solution to add iron trichloride.Add sodium borohydride (1g), this solution of vacuum concentration (water off-gas pump) adds entry (100ml) then and (3 * 150ml) extract with ethyl acetate.The organic extract liquid Na that merges ₂SO ₄Drying, the solvent vaporising under vacuum obtains crude product, and is aforementioned for another example with the chromatography crude product of purifying, obtain 3 ', 4 '-dehydrogenation vincaleucoblastine (VII, 95mg, 12%), vincaleucoblastine (315mg, 39%) and leurosidine (Leurosidine) (130mg, 16%).

In a word, characteristics of the present invention are characteristics of having described unsettled intermediate V, VI, VIII, X VI, having reached X VI a, have explained several important steps clearly, and have been different from prior art significantly.These intermediates can be separated, but there is no this necessity, and described intermediate is done careful monitoring, whole processes carried out, in a reactor shown in the reaction formula VI.

Whole yields: vincaleucoblastine (42%): white promise Centime (17%): dehydration vincaleucoblastine (18%)

Principle of the present invention, preferred embodiment, and the example of operation, as described above.Then, the limited several special shapes that are not limited to above-mentioned announcement of institute of the present invention desire protection are because of the intention of these examples is to describe unrestricted.Those skilled in the art person can carry out multiple change and change according to spirit of the present invention.

Claims (26)

1, the production method of the dimeric alkaloid compound represented of a kind of following structural formula:

In the formula:

Alk=CH ₃Or (CH ₂) _nCH ₃N=1-5 in the formula;

R ₁=CH ₃Or CHO;

R ₂=H or CO-alkyl

R ₃=H；

R ₄=COO-alkyl or CONR ₁₃R ₁₄, or middle R ₁₃And R ₁₄Be selected from the group of the aryl of the alkyl, aryl or the replacement that comprise hydrogen, alkyl, replacement;

Z=-CH=CH-or-CH ₂-CH ₂-

R=II or II a

And in the formula:

R ₇=H or COO alkyl;

R ₈=H, OH, O-alkyl, OCO-alkyl or alkyl;

R ₉=H, OH, O-alkyl, OCO-alkyl or alkyl;

R ₁₀=H, OH, O-alkyl, OCO-alkyl;

R ₁₁=H or COO-alkyl; And

R ₁₂=H or alkyl

It is characterized in that comprising the steps:

(a), by the indoles monomer with bridge nitrogen-atoms under the cooling in approximately-77 ° to about 40 ℃ of temperature, by the oxidation of bridge nitrogen, form a kind of N-oxide derivative class; And needn't separate described derivatives class;

(b), be included in acetic anhydride, fontanel in the presence of a kind of in acetic anhydride and the Acetyl Chloride 98Min. group being selected from, handle described N-oxide derivative class, to finish Pu Langnuo Paderewski (Polonovski) type scission reaction to little;

(c), product that needn't separating step (b), the product of described step (b) and indoline monomer are included in acetic anhydride, fontanel in the presence of a kind of in acetic anhydride and the Acetyl Chloride 98Min. group being selected from least, in making an appointment with-70 ℃ to about 40 ℃ low temperature, under inert conditions, carry out the stereotaxis coupling, form first imido intermediate;

(d) by each 1, described first imido intermediate is reduced in 4-dihydropyridine compound reaction, forms a kind of enamine intermediate thus;

(e) under the aeration condition of control, prepare second imido intermediate by oxidation conversion resulting described enamine compound in step (d); And

(f) product that obtains in step (e) of reduction is to form target dimeric alkaloid compound.

2, the method for claim 1, it is characterized in that further comprising described enamine that step (d) is obtained with solvent cut to original 5 to 50 times with completing steps (e).

3, the method for claim 1 is characterized in that being used for step (d), and to reduce 1,4 of described first imido-compound ,-dihydropyridine compound is to be expressed from the next:

It is characterized in that R ₁, R ₂, R ₃, R ₄, R ₅And R ₆Be independently selected from the group of the aryl of the alkyl, aryl and the replacement that comprise H, alkyl, replacement.

4, method as claimed in claim 3 is characterized in that describedly 1, and the 4-dihydropyridine is a kind of Chinese (Hantzch) ester of dwelling, R in the formula ₁, R ₂, R ₃And R ₄Be independently selected from the aryl of the alkyl, aryl and the replacement that comprise hydrogen, alkyl, replacement; And R ₃And R ₅Be carboxylicesters.

5, method as claimed in claim 3 is characterized in that the described 1.4-dihydropyridine compound of being represented by the structural formula IX comprises 1 of N-replacement, 4-dihydro-nicotinamide, R in the formula ₁Become the aryl of replacement for the alkyl that replaces; And R ₂, R ₄, R ₅And R ₆Be independently selected from and comprise: the group of the aryl of the alkyl of hydrogen, alkyl, replacement, aryl and replacement, and R ₃Be CONR ₇R ₈, in the formula: R ₇And R ₈Be selected from and comprise: the group of the aryl of the alkyl of hydrogen, alkyl, replacement, aryl and replacement.

6, method as claimed in claim 5 is characterized in that R in the formula ₁Be selected from can with positively charged imido intermediate coordination banded electron rich functional group.

7, method as claimed in claim 6 is characterized in that R in the formula ₁Be selected from the group that comprises carboxylicesters and carboxylate salt.

8, method as claimed in claim 7 is characterized in that R ₁Be 1, the 2-dimethoxy-ethyl-carbonyl.

9, method as claimed in claim 7 is characterized in that R ₁Be isobutyl--1-carboxylic acid sodium.

10, the method for claim 1, the reduction that it is characterized in that described step (d) be in inert atmosphere, in approximately-60 ℃ to about 60 ℃ temperature range, be selected from a kind of solvent at least and comprising: carry out in the group of the lower hydrocarbon of low-grade alkane alcohol, acetonitrile, dimethyl sulfoxide (DMSO), dimethyl formamide, dioxan, tetrahydrofuran (THF) and chloro.

11, method as claimed in claim 10, the reduction that it is characterized in that step (d) are to carry out in about-20 ℃ to about-60 ℃ temperature range.

12, the method for claim 1 is characterized in that oxidation switch process (e) is selected from and comprises following group:

(1) the control ventilation/oxygenation that has of described enamine solution is stirring or undertaken by this solution bubbling with air/oxygen stream in air;

(2) at described enamine and a metal ion species chosen from Fe from (Fe ^{+ 3}), cupric ion (Cu ^{+ 2}), cuprous ion (Cu ^{+ 1}), mercurous ion (Hg ^{+ 2} ₂) and silver ions (Ag ^{+ 1}) the solution of group in air, stir or by this solution bubbling control ventilation/oxygenation arranged with air/oxygen stream.

(3) at described enamine with a kind of flavocoenzyme solution stirs in air or the ventilation/oxygenation of control is arranged by this solution bubbling with air/oxygen stream.

(4) at described enamine with a kind of flavocoenzyme solution stirs in air or by this solution bubbling control ventilation/oxygenation is arranged with air/oxygen stream, wherein flavocoenzyme produce in the original place corresponding 1,5-dihydro flavocoenzyme.

(5) in described enamine and a kind of group that contains hydrogen peroxide and a kind of oxyhydroxide of representing with formula R-OOH that is selected from, wherein R is that the solution of alkyl or aryl and composition thereof stirs in air or by this solution (G) bubbling control ventilation/oxygenation arranged with air/oxygen stream, described ventilation/oxygenation be in a kind of organic solvent PH5-9 and temperature of reaction approximately-60 ℃ to the scope of+60 ℃ of pacts, carry out.

13, method as claimed in claim 12 is characterized in that oxidation switch process (e) PH carries out in the 6-8 scope.

14, method as claimed in claim 12 is characterized in that adopting about two normal iron trichlorides in step (e) (2).

15, method as claimed in claim 12 is characterized in that the aeration time at oxidation switch process (e), is from 5 to 20 minutes approximately.

16, method as claimed in claim 12 is characterized in that oxidation switch process (e) is at about 0 ℃ of row during to about 20 ℃ temperature range.

17, method as claimed in claim 2, it is characterized in that the described enamine that obtains in step (d) with a kind of solvent cut to original approximately 5 to 20 times, with completing steps (e).

18, method as claimed in claim 17 is characterized in that described enamine is diluted to 8 to 12 times.

19, the method for claim 1, it is characterized in that described reduction step (f) be temperature range from about 4 ℃ to approximately-20 ℃ and PH carry out between about 7.5 and about 8.5, and the reaction mixture in step (f) in extraction with before target compound separates, under the temperature between about 0 ℃ and about 10 ℃, vacuum concentration.

20, the method for claim 1 is characterized in that being used for step (f) and carries out reductive and comprise reaction product and a kind of being selected from from step (e) are comprised Na BH ₄, KBH ₄And LiBH ₄Basic metal hydroborates contact.

21, the method for claim 1 is characterized in that step (a)-(f) need not separate any intermediate product and operate in one pot.

22, the method for claim 1 is characterized in that must separating in advance further to react at step (c), (d) and at least a intermediate that (e) forms.

23, method as claimed in claim 22 is characterized in that separating in advance all described intermediates further to react.

24, the method for claim 1 is characterized in that this target compound is a vincaleucoblastine.

25, method as claimed in claim 24 is characterized in that further comprising that the step of the described vincaleucoblastine of oxidation is to obtain the target compound vincristine(VCR).

26, the method for claim 1 is characterized in that target compound is leurosidine (Leurosidine).