CZ20003451A3 - Novel cephalotaxane derivatives and process of their preparation - Google Patents

Abstract

The present invention relates to a new general process asymmetric hemisynthesis of harringtonines and their analogues which are alkaloids used in chemotherapy. It describes the procedure direct esterification of natural cephalotaxin with acylation a side chain precursor compound whose skeleton and functionalization are generally preformulated

Description

Technical field

The present invention relates to a process for the preparation of side-chain cephalotaxane derivatives.

BACKGROUND OF THE INVENTION

The term "cephalotaxanes" refers to compounds or salts thereof having a basic backbone of the formula

where p is equal to 1 or 2 (it is possible for 2 units to be the same or different and to be bound by a single bond or an oxygen atom) which may contain various oxygen-containing substituents (aliphatic or aromatic ethers, free or esterified alcohols, substituted or free enols and / or phenols, bridging ethers and more generally any substituent, usually occurring in the natural state on compounds of this type).

Harringtonines are alkaloids of great interest in anticancer therapy, particularly certain hematosarcomas, which are multidrug resistant to existing therapy. The selectivity of harringtonines, which is based on a novel mechanism of action related to protein synthesis, is such that this series is very promising for future use in anti-cancer therapy.

Some compilations of the literature provide a seemingly comprehensive overview of all cephalotaxane-related knowledge, and these compilations are arranged chronologically: [C.R.

mith, Jr., R.G. Powell and K.L. Mikolajzack, Cancer Treat Rep., Vol. 60, 1157 (1976); C.R. Smith, Jr., L. Kenneth, K.L. Mikolajzack and R.G. Powel in "Anticancer Agent Based on Natural Product Model," 391 (1980); Liang Huang and Zhi Xue in "The Alkaloids," Volume XXIII (ed. A. Brossi), 157 (1984); M. Suffness and G.A. Cordeli in "The Alkaloids, Chemistry and Pharmacology" (ed. A. Brossi), Vol. 25, 57-69, 295-298 (1987); P.J. 0'Dwyer, S.A. King, D.F. Hoth, M. Suffness and B. Leyland-Jones, Journal of Clinical Oncology, 1563 (1986); T. Hudlicky, L.D. Kwart and J.W. Reed, in "Alkaloid: Chemical and Biological Perspectives" (edited by S. W. Pelletier), Vol. 5, 639 (1987); M.A. Miah, T. Hudlicky and J. Reed in "The Alkaloids", Vol. 51, 199 (1998)].

Antiparasitic effects on hematozoon malaria have also been observed [J.M. Whaun and N.D. Brown, Ann. Trop. Copper. Par., Vol. 84,229 (1990)].

Homo-harrintonin (HHT) is the most active member of this series, active at doses equal to or greater than 2.5 mg / m ² body surface area in 24 hours, ie at doses 20 times lower than that of taxol.HHT has already passed fourteen trials in phase I and II and is the only known product capable of 70% reinduction of full haematological remission in patients suffering from chronic myeloid leukemia that has become resistant to alphainterferon [S. O'Brien, H. Kantarjian, M.Keating, M. Beran, C. Koler, LE

Robertson, J. Hester, M. Rios, M. Andreeff, and M. Talpaz, Blood, 332; Leukemia Insights, Vol. 3, No. 1 (1998)].

Harringtonins were extracted more than 35 years ago from exclusively Asian plants of the genus Cephalotaxacea known as Cephalotaxus harringtonia, according to a research program on new anticancer agents in the plant kingdom, developed by the National Cancer Institute. In fact, alkaloids of Cephalotaxus alkaloids consist essentially (at least 50%) of cephalotaxin, a biosynthetic precursor of harringtonines, the latter representing individually only a few percent of the total alkaloids.

In addition to their low concentration in the natural state in the plant starting material, harringtonines are mixed with a variety of congeners that have very similar chemical structures. Thus, the high performance liquid chromatography (HPLC) chromatogram of a semi-purified alkaloid extract determined no less than a few dozen cephalotaxin esters.

SUMMARY OF THE INVENTION

Taking into account that:

- on the one hand, the harringtonines are generally non-crystalline, as evidenced by the flexibility of their side chains, which are generally branched and aliphatic,

- on the other hand, these esters, especially harringtonine and homoharringtonine, are contaminated with congeners which are themselves biologically active and very difficult to separate, even by high-resolution HPLC; the state of the art does not allow these compounds to be feasible on an industrial scale for pharmaceutically active ingredients.

Although biosynthetically similar to the alkaloids of the genus Erythrina, cephalotaxanes are alkaloids having a substantially uniform structure, occurring only in the genus Cephalotaxus, the only genus in the Cephalotaxacea family. On the other hand, the side chains of the various harringtonine congeners are all derived from the primary carboxylic acid (2R) citric acid 3a methylhemiester (see appended Scheme 1) by substituting tertiary methyl using alkyl or aralkyl groups which may themselves be unsubstituted or substituted by tertiary hydroxyl groups. it then allows the later formation of a cyclic ether with a tertiary alcohol (anhydride derivatives).

The attached Scheme 1 shows the main examples of harringtonine congeners, all of which have significant cytostatic activity to varying degrees. None of the artificial analogs of cephalotaxin esters synthesized so far in the literature has at least the substructure 3b (see Scheme 1) and exhibits substantial cytostatic activity.

999997 «999997 99 99

It is useful to note that although botanically very similar to Cephalotaxaceas, Taxaceas contain triterpene alkaloids (taxins) accompanied by non-alkaloid triterpenes, taxanes, which are also substantially uniform in structure. Although quite different from taxanes in terms of chemical structures and anti-cancer mechanism of action, harringtonines are analogous to taxanes in more than one aspect:

- have cytostatic properties

- consist of a polycyclic backbone, which is an inactive biosynthetic precursor of the complete structure to which a side chain containing a similar combination of hydrophilic and hydrophobic substituents is grafted,

- the polycyclic portion of taxanes (broadcat baccans) and harringtonins (cephalotaxins) is relatively rich in the renewable parts of the plant, while the active molecules (harringtonins and taxanes) are 10 to 100 times less rich in that,

- Cephalotaxus is a rare tree, even rarer than the yew tree (Taxus) and much less abundant.

From the above, it follows that the following method of semisynthesis of taxanes by synthetic chain insertion into 10-deacetylbaccatin III of extracted origin and the asymmetric harringtonine semisynthesis by esterification of naturally occurring cephalotaxin is substantially valuable both economically and medically. Furthermore, the common incidence of Cephalotaxus is relatively reduced even at their original locus. Thus, during its import into Europe for decorative purposes in the last century, Cephalotaxus harringtonia is no longer in spontaneous form in eastern China and northern Japan. The use of the precursor present in the renewable part of the tree (leaves) for the semisynthetic preparation of homo-harringtonine is of major environmental concern, especially since the overall synthesis of optically active cephalotaxin has not yet been achieved, despite extensive synthetic studies in this regard however, the laborious syntheses of racemic cephalotaxin containing 10 to 15 degrees were performed: see biological review above).

It should be borne in mind that in order to meet the trade needs of homo-harringtonine (several kilograms per year), it is necessary to extract several hundred tons of this rare and slow-growing tree (even slower than Taxus sp.), While semisynthesis will only require several tons of renewable parts of the plant (leaves). Furthermore, homo-harringtonine (HHT) of natural origin, which is commercially available, is contaminated with congeners which, due to their structural nature, are very difficult to separate, even by high-performance preparative liquid chromatography.

First of all, since the use of cephalotaxin alone as a source for semisynthesis has not yet been economically justified, no procedure has yet been described for the selective extraction of this substance. Moreover, of the active compounds, only harringtonine and isoharringtonine are the subject of US patent applications for their preparation by extraction [R.G. Powel et al., US 3,793,454 and US 3,870,727]. Harringtonine is the subject of Japanese patent [JP 58-032 880] and deoxyharringtonine is the subject of US patent [US 3,959,312]. With respect to the preparation of homoharringtonine alone, it is the subject of only a few semi-synthetic studies [T. Hudlicky, L.D. Kwart and J.W. Reed in "Alkaloid: Chemical and Biological Perspectives" (ed. SW Pelletier), Vol. 5, 639 (1987) and J Reed in "The Alkaloids", Vol. 51, 199 (1998)], but no patent application was filed with respect to the semisynthetic process or even the extraction process.

Another aspect of the present invention is that cephalotexin can serve as a starting point for the synthesis of cephalotaxoids and harringtoids useful for anticancer (cancer and non-cancerous tumors), antiparasitic, antifungal, antiviral and antibacterial chemotherapy.

The harringtonines consist of a complex alkaloid polycyclic alcohol (cephalotaxin), esterified in the side chain, which has no greater biological activity in isolation than cephalotaxin but is essential for the biological activity of the whole. By saponification of the side chain under harsh conditions, cephalotaxin free base and harrintonic acid are obtained. The side chain attachment occurs at the end of biosynthesis. It has been demonstrated that catabolism leading to this reaction can be induced in vivo by affecting the environment or physiological stress exerted on the plant [N.E. Delfel, Phytochemistry, 403 (1980)].

• · · · · · ·

Cephalotaxin, a polycyclic moiety comprising 5 fused rings, has a novel arrangement that is unique in nature, ie, benzodioxoazepine, to which the spiropyrrolidinopentenediol system is fused. Cephalotaxan contains four asymmetric centers: three 'asymmetric carbons' and a heterocyclic tertiary nitrogen atom. The only reactive function is the secondary alcohol in position 3, the methyleneol ether in position 2 is potentially susceptible to hydrogen attack. The whole forms a pseudohelical structure enclosing the hydroxyl group in a tetrahydrazepine tube. Cephalotaxin bases readily form highly crystalline stable salts (e.g., hydrochlorides and perhydrochlorides).

This alkaloid is relatively insensitive to the basic environment. On the other hand, some authors describe a certain threshold of acid sensitivity and the quaternization of nitrogen with methyl iodide, leading to racemization by simultaneous inversion of 3 asymmetric carbons and nitrogen [D.J. Abraham, R.D. Rosensten and E.L. McGandy, Tetrahedron Letters, 4085 (1969)]. However, after a few days at pH 1-4 and at 20 ° C, this structure remains intact (personal observation).

This compound and its congeners that are not O-acylated at the 3-position are biologically inactive.

All harringtonine side chains that have substantial biological activity typically contain a 2-alkyl-2'-carbomethoxymethyl-2-hydroxyacetyl unit.

An alkyl chain of varying length has either a isopropyl-bearing branching (harringtonine HT and homoharringtonine HHT) at its end or a non-carrying (deoxy-homoharringtonine DHT) tertiary alcohol group or a phenyl group (for example, the recently isolated neoharringtonine). In the case of anhydroharringtonines, the chain may be closed by dehydration between two tertiary alcohols, for example by forming a substituted tetrahydropyran ring. The tertiary carboxyl of this complex diester is carried by the hydroxyl of cephalotaxin. A single chiral center on the side chain is located relative to the ester bond. It contains, in addition to the first secondary chain, hydroxyl which, due to its tertiary nature, has no possibility of epimerization.

• φ

Scheme 2 shows a synthetically known method for preparing harringtonines.

Several semisyntheses of natural cephalotaxin esters and several series of analogs having simplified chains but giving these analogs reduced cytotoxic activity, especially deoxyharringtonine and isoharringtonine, are described herein. Most of them relate to the simpler and less functionalized esters that form HT and HHT, esters that are very useful in chemotherapy [e.g., deoxyharringtonine, isoharringtonine, T. Hudlicky, L.D. Kwart and J.W, Reed in Alkaloid: "Chemical and Biological Perspectives" (edited by S. W. Pelletier), Vol. 639 (1987)].

All literature from 1972 to the present [Mikolajazack et al., Tetrahedron, 1995 (1972); T. Hudlicky, L.D. Kwart and J.W. Reed in "Alkaloid: Chemical and Biological Perspectives" (edited by S. W. Pelletier), Vol. 5, 639 (1987); M.A. Miah, T. Hudlicky J. Reed in "The Alkaloids", Vol. 51, p. 236 (1988)] reports the impossibility of still esterifying the highly sterically hindered secondary hydroxyl group of cephalotaxan 2a with the tertiary carboxyl of the alkanoyl moiety of harringtonic acid 3e totally converted to form harringtonins 4b. i.e. conversion 2a + 3e = 4b as described in the example shown in the diagram below.

Thus, most of the syntheses described so far involve the binding of the secondary side chain -CH ₂ CO ₂ Me, ie:

(1) Reformat reaction between methyl bromoacetate and carbonyl (real or possible) on the side chain pre-bound to cephalotaxin in the presence of zinc, or

2) prior formation of an organolithic agent.

Thus, all described syntheses include esterification of cephalotaxin using ketoalkanoyl chloride 7 lacking terminal hydroxyl and containing no secondary chain attached to the tertiary carboxyl group or any tertiary hydroxyl linked to the carboxyl group to yield 8, which is then converted to harringtonine 4a according to the reaction scheme described below.

In formula 8, CTX is a cephalotaxyl group of the formula:

It should be noted that this hydroxylation, which at the same time forms a chiral center on the side chain, never occurs asymmetrically.

Several synthetic routes include esterification of cephalotaxin with substituted hemisuccinyl chloride, optionally followed by subsequent introduction of tertiary hydroxyls (tertiary hydroxyls).

To date, O-acylation of cephalotaxin using fully pretreated and functionalized precursors to a tertiary carboxyl has not been performed [T. Hudlický, L.D. Kwart and J.W. Reed in "Alkaloid: Chemical and Biological Perspectives" (edited by S. W. Pelletier), Vol. 5, pp. 681-675 (1987); M.A. Miah, T. Hudlicky and J. Reed in "The Alkaloids," Volume 5, pp. 224-236 (1998)].

As a result, the methods of preparing harringtonines by semisynthesis, which have been described to date, have the following drawbacks:

• · · · · · «· · · · ·« · · · · · · · · · φ

- absence of stereoselectivity,

- weak convergence,

- average yields,

- functionalization and chain building on rare and expensive substrates,

- Chiral homo-harringtonine has not been obtained yet.

Since cephalotaxin is found in nature in a partially racemized form [personal observation; Huang et al., Scientia Snica, Volume XXIII, 835 (1980)], can lead to processes known in the art that use natural cephalotaxin as a starting material only theoretically to partially racemized harringtonines. Thus, the present invention has the advantage of obtaining enantiomerically pure harringtonines, even from racemic cephalotaxin, since:

(1) the asymmetric center on the side chain is formed before the esterification step, ie the side chain precursor can be obtained in enantiomerically pure form before being bound;

2) The diastereoisomers obtained in the case of racemic cafalotaxan can be separated by chromatography.

The present invention consists of:

esterification of the hindered cephalotaxin free alcohol or alternatively the corresponding metal oxide, using a chain in the form of a suitably substituted tertiary carboxylic oxacycloalkanoic acid that is completely transformed in terms of both skeleton and functionalization to prepare anhydro-homo-harringtonic acids by semi-synthesis.

opening of the resulting cyclic side chain to obtain the corresponding diols, i.e. harringtonines (as defined above).

- a description of a new preparation of all diastereoisomers of the dihydroxylated side chains of harringtonines in dehydrated cyclic form (anhydroharringtonic acid), or in which two hydroxyl groups are protected together by a bivalent protecting group to form a ring.

- differentiation of all harrington and anhydroharrington acids to separate them with cephalotaxins.

• ·

Thus, one part of the present invention consists of a synthesis, in particular anhydroharringtonine, harringtonine, anhydro-homo-harringtonine and homoharringtonine.

The present invention also relates to the esterification of cephalotaxins or their metal alkoxides with N-alkyl- and N-carbamoyl-2-alkylisoserine.

Subsequent observations and comparative studies conducted in the taxane series, irrespective of the spherical dimension, which is even larger than the hydroxyl of cephalotaxins, the hydroxyl group located at position 13 on the taxane ring allows acylation with a relatively large chain such as N-benozylphenylserine protected in the taxane. position 2 '(although attempts to acylate baccatin protected with a chain bearing a hydroxyl group protected benzoyl group failed).

Experimental studies of high-volume acylation, such as pivalic acid, showed the impossibility of acylating the hydroxyl group located at the 13-position of abaccatin protected by the dialkylcarbodiimide method, while the same reaction performed on cephalotaxins showed easy coupling of the pivaloyl chain.

It has also been demonstrated, again in the taxane series, that the improvement in the dynamic compactness of the alkanoyl or aralkanoyl chain by bifunctional cyclic protection substantially facilitates coupling. For example, French patent applications [J.P. Robin et al., FR 95/12739 and FR 95/15557] indicate that suitably protected linear N-benzoylephnylisoserines react for several hours at 80 ° C to form epimerization products, oxazoline or oxazolidine cyclic precursors react for less than one hour at 25 ° C.

Using the same conditions as above with a chain that was previously subjected to dehydration cyclization allows acylation of cephalotaxin or its alkoxides within a few hours at room temperature.

The ease of this acylation is all the more surprising since many authors have proposed syntheses that have the above drawbacks due to the impossibility of acetylation explained

ΦΦΦΦΦ · ·· ·· ··

Φ ΦΦ - - - - - - - - - - - - - - - - · · · · · · · · · · · · · · Steric hindrance at two sites: cephalotaxin hydroxyl and tertiary carboxyl side chain precursor [Mikolajzack et al., Tetrahedron, 1995 (1972)].

According to a preferred embodiment of the process of the invention, the hydroxyl function of cephalotaxane is esterified with a 2-carboxyl-2-alkyl-1-oxacycloalkane derivative.

The cephalotaxane hydroxyl whose skeleton corresponds to the general formula 1 or more preferably the secondary alcohol located at the 3-position or its metal alkoxide corresponding to the general formulas CTX-OH and CTX-OM, where M is a metal and CTX- is a cephalotaxyl group as defined above, is esterified with a tertiary carboxylic acid a function of a substituted cycloether corresponding to the general formula of type 3k or alternatively preferably one of its activated forms, isolated or formed in situ, corresponding to the general formula of type 3I to form 4c according to the reaction below:

R-O

-o (CH

- after ₂ h.

(CHJR -

CTXOH

-COA_ (CH

-O

R

3k

R. 31 or CTXOM

2'n

-CO ₂ CTX

4c

J

R ⁵ , R ⁶ and R ⁸ are independently from each other

- hydrogen,

a linear or branched and / or cyclic, saturated, unsaturated or aromatic hydrocarbon group, in particular an alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocycloalkyl group, wherein said hydrocarbon group bears or does not carry heteroatoms and R ⁶ and R ⁸ may be linked to each other to form a circle

an oxygenated ether bearing one of the above groups.

n is between 0 and 8.

A is a leaving group which is capable of leaving a carbocation, 31 may in particular be an acid halide, anhydride, mixed anhydride or cyclic anhydride (where R ⁵ = -CH 2 CO 2 -).

The type 3k free acid or any activated form of the type 3I acid can be used to esterify cephalotaxanes corresponding to formula 1, or, for example, CTX-OH type cephalotaxins. Alternatively, anhydride 3I can be prepared separately and isolated and then contacted with an alcohol. This is similar to the 3p cyclic anhydrides

wherein n, R ⁶ and R ⁸ are as defined above and which can be easily prepared from the corresponding dibasic acids to once again provide 3k esterification of methanol or alternatively coupled as above to the alcohol function of cephalotaxin type 2, but with less yield above, and the primary acid function is then conventionally methylated using methanol in the presence of a protonic acid or Lewis acid, or alternatively using a boron trifluoride etherate / methanol or diazomethane mixture.

Although less efficient and laborious, the method using acid chloride 3k provides the desired ester 4c.

All reactants of type 3k, 3I and 2 as well as the resulting esters of type 4c can be used alone or in enantiomerically pure form or in the form of a racemic mixture or in the form of diastereoisomeric mixtures. Intermediates may not be isolated in some cases, they may be formed in situ.

The reactions may be carried out at a temperature between 0 ° C and 140 ° C with or without an organic solvent, the solvent being used alone or a mixture of solvents.

The esterification of the cephalotaxane hydroxyl with a 2-carboxyl-2-alkyl-1-oxacycloalkane derivative can be carried out either by an acyl to alcohol alcohol or by a carbodiimide method.

The esterification reaction with an acyl carrier to an alcohol can advantageously be carried out according to the following six methods:

(a) esterification of the free acid with an alcohol under acid catalysis, (b) esterification of the acyl transporter via anhydrides or halides, (c) esterification of the acyl transporter using activated esters, (d) esterification with scandium triflate, (e) esterification with boron trifluoride etherate, ( f) esterification by the thioester method.

The esterification (a) is carried out by contacting a type 3k acid and a type 2 alcohol in solution in a co-solvent and in the presence of an acid catalyst. Equilibrium may be promoted by the addition of a dehydrating agent or by azeotropic removal or partitioning between two immiscible solvents, one being miscible with the ester formed and the other miscible with water. Different methods can be combined.

The catalyst may be a protonic acid such as sulfuric acid, hydrochloric acid, boric acid, preferably p-toluenesulfonic acid or a Lewis acid, which may or may not be supported by a polymer such as aluminum chloride, chlorotrimethylsilane or preferably boron trifluoride etherate. Advantageously, an ion exchange resin or hydrogen sulphate on graphite may be used.

For example, the dehydrating agent may be a dehydrating agent consisting of an inorganic salt which is inert to the reaction, such as magnesium sulfate, sodium sulfate, calcium chloride, or preferably molecular sieves.

The esterification (a) uses the same reaction conditions as above. However, in order to achieve the formation of an acyl ion characteristic of the process, conventional catalysts are replaced, for example, with anhydrous sulfuric acid or

99: · · · ·.:: ···.:. ·· • 999 · • ·. Superacids such as hydrofluoric acid and its derivatives or antimony fluoride.

The esterification (b) consists in using the same physical operating conditions as above, but using 3I, wherein A is a halogen (acid halide), another alkanoyl molecule that is identical (anhydride) or another (mixed anhydride), such as trifluoroacetyl, 2,4,6-trichlorobenzoyl, formyl, methoxyformyl, sulfonates, phosphates and chlorophosphates.

In a variant of the above method and in the specific case where R ⁵ = CH 2 CO 2 H, cyclic anhydride of type 3p may be used

wherein R ⁶ and R ⁸ are as defined above, which can be prepared very simply by treating the corresponding dibasic acid with acetic anhydride, for example under the mixed anhydride operating conditions described below.

The catalysis may be acidic as mentioned above or preferably alkaline, for example a tertiary base such as pyridine and / or dimethylaminopyridine (DMAP), pyrrolidinopyridine (PPY), triethylamine or a strong base such as a hydride, for example calcium hydride. The solvent may be an aprotic solvent, for example hexane, toluene, dichloromethane, tetrahydrofuran and / or pyridine.

Esterification (c) is a method similar to esterification (b). These ester preparation methods can be advantageously used and include, for example, 1-acyloxy-1,2,3-triazole or formamidinium or silyl ester or 2-acyloxypyridinium intermediates.

• 9 · 9 · 9 · 9 · · · · · ·

The carbonyldiimidazole method can also be used, wherein the Nacylimidazole intermediate can be used.

The carbodiimide method uses a dehydrating coupling agent such as carbodiimide, for example, dicyclohexylcarbodiimide (DCC), 1,3-diisopropylcarbodiimide (DIC) or 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide.

The reaction may be catalyzed, for example, by a tertiary base, such as pyridine and / or dimethylaminopyridine (DMAP), pyrrolidine pyridine (PPY), triethylamine, 4-morpholinopyridine or any suitable base. Nhydroxybenzotriazole (HOBt), N-hydroxysuccinimide (HO-Su) may also be used.

The molar ratio of 3I to 2 may be between 171 and 4/1.

The reaction can advantageously be carried out under an inert gas at a pressure close to atmospheric pressure, preferably at a temperature between 0 ° C and 110 ° C.

The solvents used are preferably organic solvents such as toluene and / or dichloromethane and / or chloroform and / or tetrahydrofuran and / or dimethylformamide.

The application of the methods described above for substituted carboxylic oxacycloalkanes to coupling their synthetic precursors of (linear) 1-hydroxy-1-methoxycarbonylmethylalkenecarboxylic acids makes it possible, contrary to all expectations, to synthesize type 4c esters in one step from type 3f ethylenic tertiary hydroxy acids instead of 3f, 3k, 31, 4c.

where m is between 0 and 3,

- ¹ 4c w in formula 4c, n, R ⁵ , R ⁶ , R ⁸ and CTX has the same meaning as above.

Indeed, the ethylene tertiary hydroxy acid 3f treated under conditions similar to the above conditions directly provides the cyclic ester of cephalotaxins of type 4c without isolation of the intermediate. In the case of the mixed anhydride 31 method, in situ formation of the corresponding lactone can be assumed based on the presence of an infrared absorption band at 1840 cm ^-1 .

These bases were confirmed by formation and isolation of 3I using 3f only under the activation conditions described above, i.e. the formation of mixed anhydride, for example in the presence of 2,4,6-trichlorobenzoyl chloride or alternatively, in the so-called DCC method described above.

Substituted carboxylic cycloethers of type 3k, substituted ethylene tertiary acids of type 3f, their activated intermediates, and cyclic anhydrides of type 3p may be coupled with cephalotaxins, either in a racemic series or more preferably in an optically active series.

In the case of coupling between one of the above types of acids in the form of racemic mixtures, with one enantiomer of cephalotaxin, relative stereoselectivity was found due to chirality and marked steric hindrance at the reaction site in the sense that the ratio between each of the two diastereoisomers is usually different than 1 .

Separation of the two resulting type 4c diastereoisomers (R ⁵ = CH 2 CO 2 Me) known as "anhydroharringtonine"

 · «____ ____ ____ ____ ____ ____ ____ ____ ____ ____ ____ ____ ____ ____ ____ ____ ____ ____ ____ ____ ____ Where n, R, R and CTX have the same meaning as above, preparative chromatography can be carried out either in the so-called normal phase, for example on natural silica gel as the stationary phase and mixtures of organic solvents as the mobile phase or preferably in reverse phase, for example, inert silica grafted with apolar groups such as organosilyl, cyanoalkyl, phenylalkyl, preferably octadecylalkylsilane, and mixtures of aqueous solvents as the mobile phase.

In the case of enantiospecific coupling, no secondary epimerization of any parent moiety is observed and the single diastereoisomer obtained can be crystallized. If this diastereoisomer does not crystallize, it is subjected to flash chromatography to remove reaction residues and then precipitated by addition to a non-solvent to be isolated as an amorphous powder.

The opening of cephalotaxin carboxylate cycloesters of type 4c to obtain the haloalcohol 4d is shown in the following scheme.

R '

4c

CO.CTX

X OH - (CH ₂ ) J-CO ₂ CTX

R ^e R ⁵

4d

In these formulas, n, R ⁵ , R ⁶ , R ⁸ and CTX have the same meanings as above, X being a heteroatom as halogen.

Cyclic ethers of type 4c may in some examples have the particular feature that they simultaneously contain methylene ol ether, for example in the 2-position of the cephalotaxins. Despite the usual inertia of true cyclic ethers (i.e., non-hemiacetal ethers), contacting solution 4c in an organic solvent, preferably a chlorinated solvent such as dichloromethane or chloroform under controlled conditions, i.e. in the presence of, for example, dilute hydrochloric acid, preferably hydrobromic acid at low temperature or additionally in the presence of halo-trialkylsilane or alternatively boron trihalide, for example boron tribromide at low temperature in an organic solvent, preferably a chlorinated solvent such as dichloromethane, dichloroethane or chloroform,

IA allows selective opening to give a haloalcohol of type 4d, such that X = halogen, in quantitative yield without any observable formation of the demethylation product, even if the cephalotaxins carry enol ethers or other functions sensitive to ether cleavage agents. In any case, even in the case of accidental demethylation, the enol can be readily selectively remethylated as described in the literature (for example, by dissolving in methanol in the presence of ptoluenesulfonic acid).

Haloalcohols of type 4d where X = halogen are of great interest as substrates necessarily leading to heteroatom analogs, in view of the very good reactivity to the halogen substitution they carry.

Another variant consists in contacting the product 4c with an aqueous acid, optionally in a miscible or immiscible co-solvent. The acid may be, for example, a protonic acid, in which case it is an organic or inorganic acid, preferably hydrochloric acid, tetrafluoroboric acid or formic acid. In this case, diols of type 4b can be isolated directly without passing through the haloalcohol stage described above.

Halogen alcohols type 4d, where X = halogen are hydrolyzed according to the following scheme:

OH

OH OH ⁸ - (CH 2)

2'n

CO, CTX (CH ₂ ) _n ·

- CO, CTX

R R

4d wherein n, R ⁵ , R ⁶ , R ⁶ and CTX have the same meaning as above.

Type 4d haloalcohols are particularly preferred for controlled hydrolysis by contacting with an aqueous inorganic base, such as dilute sodium hydroxide, sodium carbonate, or preferably barium carbonate, at a temperature between 0 ° C and 30 ° C, with stirring to give type 4b diols .

»· · ·

One variant is in situ hydrolysis of the intermediate type 4d halogenated alcohol at the end of the reaction, preferably accomplished by treating the cyclic type 4c ether in dichloromethane in the presence of hydrobromic acid in acetic acid and directly adding water or hydrolysis solution to the reaction mixture under cooling and vigorous stirring.

The process according to the invention is particularly suitable for the preparation of azaharringtonines, nitrogen analogues of harringtonines.

In order to demonstrate, for example, the flexibility of using haloalcohols 4d where X = halogen as substrates, such materials were, for example, subjected to azidolysis by treatment using, for example, an alkali azide in a solvent such as ethanol, methanol or dimethylformamide. such as an alkanol or a lower ester leads to the corresponding aminoalcohol corresponding to formula 4f (aminodeoxyharringtonine). The amine can be amidated under Schotten-Bauman conditions, i.e., in an aqueous medium in the presence of an inorganic base catalyst, to give the amidoalcohol type 4e.

X OH NH, | OH R * - ch -L-c-ctx -R * —UcH ₂ ) _n —CO ₂ CTX r ‘ R ^s r ‘ R ³

OH

- (CH

2'n

R

4e

Wherein Z = NHCOR or NHCOAr, R and Ar are as defined above, more preferably alkyl or aryl groups which may be optionally substituted. The amine 4f may be sec-alkylated to form an alkylaminodeoxyharringtonine (Z = NHR or Z '= NHAr or Z = NR ₂ or Z = NHAr ₂ , wherein it is possible for two R and Ar groups to be identical or independent) or acylated to form amides (Z = NHCOR or Z = NHCOAr) or carbamates (Z = NHCOOR) derived from the corresponding aminodeoxyharringtonine 4f, n, R ⁵ , R ⁶ , R ⁸ , R ⁹ , X and CTX are as defined above.

Alternatively, cyclic ethers of type 4c may be suitable for the Ritter reaction in the presence of a nitrile (which may serve as a solvent) at a low temperature between -100

° C and + 30 ° C in the presence of an acid such as sulfuric acid, perchloric acid or preferably tetrafluoroboric acid, to obtain acylaminodeoxyharringtonine (Z = NHCOR or Z = NHCOAr), derived from the corresponding aminodeoxyharringtonine 4f.

Extraction of cephalotaxins type 2 is carried out according to the procedure below.

Cephalotaxins of type 2 can be prepared according to methods described in the literature, either by synthesis or by extraction. In the case of extraction, since none of the methods uses direct contact of the plant starting material with aqueous acid, it has been found advantageous to describe this method in the present invention. The fresh or dry plant material is contacted for 24 hours with the acidified aqueous organic mixture using a dilute inorganic acid or a weak organic acid such that the pH is between 1 and 4, preferably 3. The inorganic acid may be, for example, sulfuric acid or hydrochloric acid and organic the acid may be citric acid, lactic acid or tartaric acid; the organic solvent may be, for example, a lower alcohol, a ketone, tetrahydrofuran or any water-miscible solvent used by a person skilled in the extraction. The water content is between 20 and 80%, preferably 50%. The solution obtained can be directly chromatographed or alkalinized in order to be countercurrently extracted since, unlike the methods described in the literature, it contains no chlorophyll and / or vegetable fat. Countercurrent extraction using a water immiscible organic solvent such as a lower ester or, preferably a halogenated hydrocarbon, more preferably dichloromethane, provides a mixture of all alkaloids as a white powder. There are several methods for the purification of Cephalotaxus alkaloids, but none of them are particularly adapted to the purification of cephalotaxins and in particular the cephalotaxins of formula 2a, especially in the reverse phase.

The present process for the purification of cephalotaxins, which forms an integral part of the new process for the semisynthesis of harringtonines, thus includes reverse phase chromatography, which has not yet been described for these purposes. Reverse phase chromatography uses as stationary phase, for example, inert S1O2 grafted with apolar groups such as organosilyl, cyanoalkyl, phenylalkyl or preferably octadecylalkylsilane chains, such as those encountered in commercial use, and a mixture of aqueous solvents such as the mobile phase, water alone (without organic

solvents); The pH is adjusted, i.e., to a value between 2 and 4 using inorganic acids such as hydrochloric acid or sulfuric acid. It is also preferred to add an additive such as aqueous ammonia or triethylamine. According to this process, which is economically very advantageous since it does not use an organic solvent and allows the stationary phase to be reused for virtually hundreds of operations, cephalotaxin is obtained in quantitative yield and with a purity of more than 95%.

The above process makes it possible to obtain not only the levorotatory cephalotaxins occurring in the plant material, but also the racemic cephalotaxins which are also present in the natural state.

Metal alkoxides corresponding to the general formula 1b (n = 1 to 12), wherein M is a metal, in particular an alkali metal such as sodium, potassium or lithium or a transition metal, for example zirconium, titanium or zinc, can be obtained by metalating one or more hydroxyls in mono- or polyhydroxycephalotaxanes corresponding to the general formula 1a (x = 1 to 12) and wherein M is more preferably an alkali or alkaline earth metal or any metal which may form an alkoxide.

This formation of alkoxides is of great value for facilitating the acylation of hindered hydroxyls of cephalotaxanes, most preferably in the coupling of this cephalotaxane with acylating side chain precursors, automatically leading to the harringtonines of the present invention.

Some methods can be used to metallate hydroxyls of cephalotaxan. For example, a metal hydride, an alkyl metal, an amide or, more generally, any agent capable of exchanging or providing a metal atom may be used.

Simple contacting of the hydroxylated cephalotaxan in an organic solution, preferably under an inert gas, with a metal hydride such as potassium hydride, lithium hydride or more preferably sodium hydride leads to a metal alkoxide of cephalotaxan which may, for example, serve as an in situ substrate to bind e.g. a suitably substituted alkyl, acyl or alkylsilyl group. The organic solvent may be a suitable aprotic solvent such as an ether, more preferably tetrahydrofuran, a liquid aromatic hydrocarbon, preferably toluene, or more preferably any organic solvent that is liquid at the temperature and pressure conditions used and which has no appreciable reactivity to the reactant. The temperature of the reaction medium may be between -90 ° C and +30 ° C.

Simply contacting the hydroxylated cephalotaxane in an organic solution, preferably under an inert gas, with a metalized hydrocarbon such as a lithiated hydrocarbon, preferably butyllithium, leads to the same alkoxides as those mentioned above. The same solvents as above can be used except that the reactivity of the metal hydrocarbons is generally greater than the reactivity of the metal hydrides, at a temperature between -100 ° C and -20 ° C, preferably between -60 ° C and -80 ° C.

The simple contacting of the hydroxylated cephalotaxane in an organic solution, preferably under an inert gas, with an amide, preferably an alkali metal amide, for example an alkali metal dialkylamide such as lithium dicyclohexylamide or lithium diisopropylamide or an alkali metal (lithium, sodium or potassium) amide to the same metal alkoxides as above. The same solvents used may be used

2a

By way of example, without departing from the generality of the present invention, cephalotaxin 2a in a stirred tetrahydrofuran solution at -70 ° C is treated with one equivalent of butyllithium or bis (trimethylsilyl) amide and is treated within a few hours.

• ·

yields lithium alkoxide 2h which, when collected in situ with acetic anhydride, provides 3-O-acetylcephalotaxin 2b.

The preparation of substituted racemic carboxylic cycloethers of type 3k is described in detail below.

According to a first variant, the preparation of the following carboxylic cycloethers can be carried out by cyclization of the substituted ethylene tertiary hydroxy acid type 3f according to the following scheme:

COjH

R-O

R • o (CH

2'n

-co ₂ a

R

COjH

R

3k wherein m, R, R ^5, R ^6, R ⁸ and A are as defined above.

AND

Substituted carboxylic cycloethers of the type 3k, such as, for example, of formula A or

B:

may be prepared from substituted ethylene tertiary acids of type 3f by simple dissolution in an organic solvent, in the presence of an acid.

According to a second variant, the preparation of these carboxylic cycloethers can be carried out by cyclization to form acyl compounds. As mentioned above, acids whose tertiary alcohol group is free, such as 3f, are cyclized spontaneously by treatment with the dehydrating agent required for certain cephalotaxin alcohol acylation techniques mentioned above and then esterifying the latter to give 4c.

In the absence of the alcohol substrate, the ethylene tertiary alcohol type 3f leads to the isolation of the acyl intermediate 31 above or, after hydrolysis, to the isolation of the type 3k acid under anhydrous working conditions.

In this case, the procedure used is exactly that described for coupling involving the formation of acyl compounds in situ but in the absence of a cephalotaxin-type substrate.

According to a third variant, these substituted 3-carboxylic cycloether ethers can be prepared by deprotecting the tertiary carboxyl of a suitably substituted 3h precursor,

RO (CHA-co ₂ gp

3k

R ⁵

3h

R ⁵ , R ⁸ , R ^{6 and} n are as defined above and GP is an acid protecting group, in the specific case GP = R.

According to a fourth variant, in a specific case in which R ⁵ = CH 2 CO ² R, suitably substituted carbocyclic cycloethers of type 3k, such as where R ⁵ = CH 2 CO ² R below,

R (CH ₂ ) -CO ₂ H

CH ₂ CO ₂ Me

3k

R ⁸ , R ⁶ and as defined above may be prepared by total saponification of the corresponding diesters 3j, such as where R ⁵ = CH 2 CO ² R, followed by fine selective methylation of the dibasic acid 3r, where R ⁵ = CH 2 CO 2 H.

R ⁸ .

RR *

RO co ₂ y

CH ₂ CO ₂ Me (CH ₂ ) (CH ₂ ), -, - ° ^C 2 ^H CH ₂ CO ₂ H 3r

3k t

3e

-CO ₂ R

CH ₂ CO ₂ H

3t

This process, carried out at room temperature and under close monitoring of the reaction kinetics, results in the selective saponification of the above primary ester; to give derivatives of the 3t type, wherein R = GP is furthermore, (CH 2 O)

--CO ₂ GP ch ₂ every ₂ h

3t (R = GP)

R ⁸ , R ⁶ , GP and n are as defined above which can be coupled with cephalotaxins using the methods described above to ensure the absence of transesterification during coupling of the tertiary acids of the present invention.

According to a fifth variant, the preparation of these carboxylic cycloethers can be carried out by regioselective methanolysis of the corresponding cyclic anhydride.

As mentioned above, the dibasic acid 3r is spontaneously dehydrated to the cyclic anhydride 3p, which is a good acylating agent for alcohols, so that 3k methanolysis also advantageously yields a product where R ⁵ = CFhCChMe, which constitutes another method of preparation.

The preparation of substituted ethylene tertiary hydroxy acids of type 3f is described in detail below.

Substituted ethylene tertiary dibasic hydroxy acids, which are precursors of type 3f monoacids, can be obtained, like their cyclic analogues of type 3k from above:

- either by selective deprotection of their type 3i precursors

or, for example, in the case of dibasic acids of the type 3s, wherein R ⁵ = CH 2 CO 2 H,

_{CH2 CO2} H, selective methylation of the primary carboxyl groups, in particular prolonged contact of the reactants with a methanol solution at room temperature using a boron trifluoride / methanol.

Dibasic acids of type 3s, where R ⁵ = CH 2 CO 2 H may be obtained by saponification of the corresponding diesters of type 3q, where R ² = ChkCCUMe, by contacting the latter with an excess of the base in an aqueous or aqueous-alcoholic medium, r ⁸

co ₂ r

Zs

co ₂ h

COjH

3s

co ₂ h

CO ₂ Me

3f m, R, R ⁸ and R ⁶ are as defined above.

The alcohol may be a lower alcohol such as methanol, ethanol or preferably isopropanol, and the base may be, for example, an alkali metal or an alkaline earth metal or a rare earth metal hydroxide or aqueous ammonia. If the reaction is carried out at a temperature between 0 ° C and 30 ° C for 15 minutes to 1 hour, the regioselective saponification of the primary ester can proceed without difficulty. By raising the reaction temperature to the boiling point of the solvent mixture and / or prolonging the reaction time, the dibasic acid is obtained in good yield without the formation of by-products.

The dibasic acids of the 3s type, where R ⁵ = CH 2 CO 2 H, due to their crystallogenic properties, can then generally be obtained in enantiomerically pure form by subsequent crystallization of the enantiomerically enriched mixture until a constant optical rotation is obtained.

The dibasic acids obtained above can then be converted to substituted ethylene tertiary hydroxy acids of type 3f, wherein R ⁵ = CH 2 CO 4 Me by selective monoesterification of their primary carboxyl group using methanol in the presence of a protonic acid or Lewis acid or alternatively using boron trifluoride / methanol or diazomethane.

As for cyclic analogs of type 3t, where R ⁵ = CH ₂ CO ₂ H from above, this process carried out at room temperature and strictly following the reaction kinetics results in selective saponification of the above-mentioned primary ester; to obtain derivatives of type 3u, wherein R ⁵ = CH 2 CO 2 H, which can be coupled with cephalotaxins using the methods described above, to ensure the absence of transesterification during the coupling of tertiary acids forming the subject of the present invention.

Substituted ethylene tertiary hydroxyesters of type 3q can be prepared according to the scheme below:

co ₂ r

RSMXor R ^S M

co ₂ r

R ^6, R ⁸ and R ⁵ have the same meaning as above.

Ethylene esters of type 3 q can be prepared according to a number of methods described in the literature for similar cases, such as the hydroxyalkylation of the corresponding 1alkyl- or 1-alkenyl-1-ketoester type 9.

By way of example and not limitation of the generality of the present invention, hydroxyalkylation of 1-alkenyl-1-ketoester of the type with lithium methoxycarbonylmethylenolate (R ⁵ M = MeOCOCHLi) or the corresponding organosinc reagent (Reformat reaction where R ⁵ MX = MeOCOChkZnBr) leads to diesters ⁵ = ChbCO 2 Me.

• ·

The same reactions applied to the chiral ester (R = R) result in a mixture of separable diastereoisomers which, upon deprotection of the tertiary acid function, lead to a diasteroisomeric pair.

In addition, the hydroxyalkylation reaction of 1-alkyl- or 1-alkenyl-1-ketoester type 9, performed in the presence of a chiral reagent such as sparteine or quinine, can provide significant enantiomeric enrichment, which can be further enhanced by fractional crystallization.

Type 9 ketoesters alone are conventionally prepared by C-semiacylation of the carbanion of the corresponding type 10 alkyl or alkenyl halides with dialkyl oxalate.

One of the advantages over the synthetic process used in the prior art that forms the present invention lies in the possibility of coupling a completely pre-formed chain with cephalotaxins. Thus, the preparation of the above anhydroharringtonic acids in enantiomerically pure 3k form is of interest since the postcopulation formation of the chiral center at the 2 'position of the harringtonines during the attachment of the secondary chain as described in the prior art leads on the one hand to an epimeric mixture on the other hand, to the loss of about 50% of the very precious cephalotaxins (not recyclable in the process for preparing medicinal substances using good manufacturing practice).

A number of methods are used to achieve this. They all use both type 3k cyclic monohydric acids or their type 3r dibasic precursors and their type 3f ethylene linear precursors, and it is also possible to apply chiral chromatographic methods to precursors that have no group capable of providing reversible chemical bonding with chiral compounds (in this case free acid function).

According to the first step of the process for the enantiospecific preparation of these acids, the epimeric mixture is formed by combination with a chiral alcohol or amine.

Reactions for esterification of the impeded secondary alcohol function of cephalotaxin with oxacycloalkanecarboxylic acids type 3k from above (including those which are

QQ · ··· ··········· · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · in situ from ethylene tertiary hydroxy acids of type 3f) may also be used to esterify another chiral alcohol to convert a racemic mixture or one of the partially enantiomerically enriched type 3k acids to a diastereoisomeric mixture to which all non-chiral separation methods can be used. The above methods are also applicable without modification to the amidation of chiral primary or secondary amines.

Thus, when oxacycloalkanecarboxylic acids of type 3k or their ethylene linear precursors react with a chiral alcohol designated ROH or an amide R 'R NH (it is possible for R' * to be replaced by hydrogen), two chemical compounds are obtained which have physicochemical properties different (e.g., NMR, melting point, solubility, chromatographic properties, enzymatic or microbiological attack, etc.). The alcohol or amide must preferably be hindered and carry its chiral center at the point of attachment to the tertiary carboxylic acid of type 3k. The alcohol may be, for example, menthol, borneol, valinol or preferably quinine. The amine may be, for example, ephedrine, more generally any commercial chiral alcohol or amide may be used.

R-O

R-O

R (CH ₂ ) -CO ₃ H (CH ₂ ) -CO ₂ AND ^R 1 ^OH (CH ₃ -O)

-co ₂ r *

R

3k

R

3 / (R = R 6) n, R ⁵ , R ⁶ , R ⁸ and A have the same meaning as above, R * has the same meaning as R, but is chiral.

By way of example, but without limiting the generality of the present invention, (-) - quinine, which, like cephalotaxins, is an alkaloid with a sterically hindered secondary alcohol function, reacts with a racemic mixture? oxacycloalkanecarboxylic acid of type 3k to give a mixture of two corresponding epimers 3v, where R ⁵ = CH ₂ CO ₂ Me and 3w, where R ⁵ = CH ₂ CO ₂ Me:

The binary epimer mixtures obtained in combination with the chiral compound can be separated, for example, by fractional crystallization, distillation, liquid-liquid countercurrent separation and, in view of the high degree of added value of these intermediates, by any general preparative chromatography, for example normal phase chromatography, size exclusion chromatography, preferably reverse phase chromatography. or grafted phase. Since these methods are synergistic, they can advantageously be combined to improve diastereoisomeric purity.

By way of example, but without limiting the generality of the present invention, a mixture of two epimers 3v (where n = 3; R ⁶ = R ⁸ = Me; R ⁵ = ChbCO 2 Me) and 3w (where n = 3; R ⁶ = R ⁸ = Me) R ⁵ = ChhCO 3 (Me) cited in the above example was separated without problems and in quantitative yield using an octadecylsilane-type graft phase and a methanol / water mobile phase.

Regeneration of type 3k oxacycloalkanecarboxylic acids in enantiomerically pure form may be accomplished by complete hydrolysis followed by selective remethylation of the primary carboxyl group of the appropriately selected diastereoisomer (see above sequence 3j (3k), or with respect to ester bond with oxygen at the benzylic position) In the latter type, the disadvantage of hydrogenolysis is largely offset by economic considerations.

By way of example, but without limiting the generality of the present invention, (-) - quinine (2'R) -anhydroharringtonate 3v (where n = 3; R ⁶ = R ⁸ = Me; R ⁵ = ChhCO 2 Me) provides ·· ·· • ··

(2R) -anhydroharringtonic acid of 3k type and dihydrodeoxyquinoline, which cannot be recycled, but this is a small disadvantage due to the low cost of this alkaloid. Alternatively, double saponification of 3v followed by selective remethylation provides a product that is completely identical to (2R) -anhydroharringtonic acid of type 3k above.

An enantiomer of the non-natural configuration (2S), after undergoing the same conversion as its (2R) enantiomer, can be used to investigate structure-activity relationships.

According to the first step of the process for the enantiospecific preparation of these acids, the racemic mixtures can be resolved by salt formation with chiral basic compounds.

Racemic mixtures of oxacycloalkanoic carboxylic acids of type 3k (including those formed in situ from ethylene tertiary acids of type 3f) can form salts with a chiral amine by simply placing them together in solution in an organic solvent. Although most of the methods described above are useful for separating esters and amides formed with 3k, for example, chromatography, since the salts formed are generally crystalline, fractional crystallization is used as a preferred method for separating 3k acids. The solvents used, alone or as a mixture, may preferably be polar organic solvents, which may or may not be combined with water, such as ketones, alcohols and lower esters. The salt-forming reaction is preferably carried out at a temperature between 0 and 100 ° C. Recrystallization can be accomplished by redissolving the salt in a mixture whose strength can be precisely adjusted by the combination of solvents and temperature used, according to standard techniques known to those skilled in the art. When diastereoisomeric enrichment is considered sufficient, the salt is decomposed in the presence of, for example, dilute aqueous acid such as hydrochloric acid or sulfuric acid. Extraction of the enantiomer of the regenerated acid may be performed using a water-immiscible solvent such as a lower ester.

By way of example, but without limiting the generality of the present invention, a racemic mixture of (2R or 2S) -anhydroharringtonic acids of type 3k may be resolved

for example by contacting (-) - ephedrine and then traction recrystallization in ethyl acetate / methanol.

The 2R-anhydroharringtonic acid of type 3k can then be regenerated by contacting the purified salt with 2N hydrochloric acid and continuously extracting the acidic aqueous phase with ethyl acetate.

The oxa-cycloalkanecarboxylic acids of type 3k described above may be subjected to preparative chiral chromatography.

The final products are purified by HPLC to obtain the final products for pharmaceutical purposes.

Despite the efficiency levels of modern methods of synthesis, semisynthesis and isolation of natural substances, it has now been established by the health organizations of industrialized countries that if the presence of impurities is greater than 1: 1000 (0.1% w / w), considered harmful to the patient.

In addition, identification followed by toxicological studies on any toxicologically unknown substance exceeding this threshold is required by law to be able to submit documentation for authorization to market medicinal products.

In addition, diastereoisomeric purity (in a specific case, enantiomeric purity) may lead to therapeutic variations; for example, it is well known that quinine (see formula above) is an anti-malaria agent, while one of its diastereoisomers is a cardiac antifibrillant.

In the therapeutic use of the compounds of the present invention, we often encounter 10 to 100 times the amount of active agent or side effects caused by changes (here involuntary) in the molecular structure.

Among the methods for achieving the desired level of purity, high-resolution industrial chromatography takes first place, its high cost being balanced by the high added value of the top active ingredients, particularly with regard to the quality it provides and the safety it offers. users.

By way of example, but without limiting the generality of the present invention, homoharringtonine 4b, (wherein n = 3; R ⁶ = R ⁸ = Me; R ⁵ = ChkCO ⁵ Me, CTX = cephalotaxyl),

OH OH

-HCH ₂ U? Co ₂ ctx

R ^'R

OH

OH • il ·

R (CH ₂ ) 2 CO ₂ CTX

Ø ⁵ (2'R) -Homoharringtonin 4b (2'S) -Epihomoharrington »n

R ⁶ , R ⁸ , R ⁵ , on CTX as defined above, can be separated from its epimer along with its impurities by preparative reverse phase chromatography using an octadecylsilane type graft reverse phase as a stationary phase and a suitably treated methanol / water mixture as mobile phase.

This process provides a product wherein the sum of related impurities is less than 0.5%, with none of the impurities individually exceeding 0.1%.

φ · · • • φ • · · · · · · · ·

SUMMARY OF THE INVENTION

The present invention relates to a process for the preparation of side chain cephalotaxan of the following formula and / or a salt thereof •-CO-O-CTX wherein Ω ("omega") is a representative terminal group of the chain moiety and -CO- is a carbonyl ester group bonded to cephalotaxan ; Ω-CO- group corresponds to:

- either of the following substituted heterocycloalkane of the general formula:

R (° ^Η Λ -cokde n is between 0 and 8;

Z is an oxygen, nitrogen or sulfur heteroatom;

R ⁵ , R ⁶ and R ⁸ are independently hydrogen;

a hydrocarbon group, saturated, unsaturated or aromatic, linear or branched and / or cyclic, in particular alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocycloalkyl, said group including or not including heteroatom (s); R ⁶ and R ⁸ may be incorporated into the cycle; an oxygenated ether bearing one of the above groups;

- or the following linear alkene of formula:

R-

ΦΦΦΦ φφ φφ ·· · φ φφφφ φφφφ φφφ φ φ φφφφ φφ φ φφφφφφ φ φ φ φφφφ φφφ φφφ φφφφ φφ φφ wherein m is between 1 and 8, R ^5, R ⁶ and R ⁸ are as defined above,

- or the following general formula

wherein n, R ⁵ , R ⁶ and R ⁸ are as defined above;

Z and Q ² are independently oxygen, nitrogen or sulfur heteroatoms;

Q ¹ is a carbon, silicon, or phosphorus atom;

R ⁹ and R ¹⁰ are independently hydrogen, alkoxy, a hydrocarbon group, whether or not including heteroarom (s), saturated, unsaturated or aromatic, linear or branched and / or cyclic, especially alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocycloalkyl ;

R ⁹ and / or R ¹⁰ may be zero or together form a heteroatom and / or form a ^1- bond with Q ¹ , R ⁹ and R ¹¹ may be zero or form between two carbon atoms carrying them a multiple bond; and

R ¹¹ is hydrogen, arylcarbonyl, alkoxycarbonyl, aryloxycarbonyl or alkylcarbonyl;

where

-Ο-CTX is the cephalotaxin moiety of the following formula, a salt thereof:

wherein p is 1 or 2;

the two types of groups -Ω and -CTX mentioned above are linked by an ester bond -CO-O36 ···· φ

• · ·

9 where the procedure includes contacting:

either carboxylic acids of the formula Ω-CO-OH or salts thereof;

or an active acid form of the formula Ω-CO-A or a salt thereof, wherein Ω-CO has the following general formula

R <CH ₂ > n -ocde n, Z, R ⁵ , R ⁶ and R ⁸ are as defined above; where Ω-CO has the following general formula:

R 2 H 0 (CH where m is between 1 and 8, Z, R ⁵ , R ⁶ and R ⁸ are as defined above; wherein Ω-CO has the following general formula:

and wherein n, Z, Q ¹ , Q ² , R ⁵ , R ⁶ , R ⁸ , R ⁹ , R ¹⁰ and R ¹¹ are as defined above A means:

a cyclic anhydride of the following general formula:

O • ···· ·· <· ·· · '* · · · · · • * «· · · •» «• ···· · · · · · wherein n, R ⁶ and R ⁸ are as defined above;

wherein the primary carboxyl group of the above compounds is methylated with:

either cephalotaxan or a salt thereof carrying at least one free hydroxyl group of the formula H-O-CTX, wherein CTX is as defined above;

or a metal alkoxide of the formula M-O-CTX, wherein GTX is as defined above and M is a metal;

or an active form of a hydroxyl group of the formula YO-CTX, wherein -O-CTX is as defined above and Y is either a leaving group to produce a negative charge on the oxygen atom by cleavage between Y- and -O-CTX, or allows carbocation by cleavage between YO - and -CTX;

optionally in the presence of one or more reaction additives to form a side chain cephalotaxan and / or a salt thereof.

Most preferably, Z is an oxygen atom and the cephalotaxan H-O-CTX is cephalotaxin of the following formula or a salt thereof:

wherein R ^1, R ^2, R ³ and R ⁴ are independently hydrogen, hydroxyl group or alkoxide.

Cephalotaxan HO-CTX as defined above is cephalotaxin or a salt thereof, wherein R ¹ is hydroxyl, R ² is methoxy, R ³ and R ⁴ are hydrogen.

• · <

o c

MeO

R ⁵ is preferably hydrogen or -CH ₂ -CO-O-Me.

The Ω-CO group is preferably such that n = 1 to 4, R ⁶ and R ⁸ are methyl.

The Ω-CO group may also be such that n is 1 or 2, R ⁶ is phenyl and R ⁸ is hydrogen.

If R ⁵ is -CH 2 -CO-O-Me, R ¹ = OH, R ² = OMe, R ³ = R ⁴ = H, the cephalotaxane is preferably such that n = 0, Z is a nitrogen atom and R ⁸ is hydrogen.

A may be a Ω-CO-CO group where Ω is as defined above, or the halide A may also be a group of a Ω-CO-A compound having the ability to generate a bond cleavage between a carbonyl group and a substituent A to provide Ω-CO * and A ’.

Further, A is a group selected from the substituents:

methoxyformyloxy of formula MeOCOOtrifluoroacetyloxy of formula CF 3 COO-, alkylsulfonyloxy of formula RSO ₂ -, phosphoxy of formula (RO) ₂ POhalophosphoxy of formula ROP (CI) Otrialkylsilyloxy of formula R 3 S 10 -, wherein R is alkyl, dimethylformamidium chloride of formula

JVIe

Or acyloxypyridinium bromide of the formula

Br

A may be 2,4,6-trichlorobenzyloxy or a group corresponding to the following formula:

In the case where A is carbonylimidazole, where A is 2,4,6-trichlorobenzoyloxy, the reactant of the formula Ω-CO-A is obtained by contacting the Ω-CO-OH acid as defined above with 2,4,6- trichlorobenzoyloxycarbonyldiimidazole in the presence of a strong base such as an alkoxide

According to the carbodiimide method, the coupling additive is a substituted carbodiimide and / or a basic additive, such as a tertiary amine.

For example, substituted carbodiimide is selected from cyclohexylcarbodiimide (DCC), 1,3-diisopropylcarbodiimide (DIC), and 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide chlorohydrate.

A cephalotaxin alkoxide corresponding to the general formula M-O-CTX, where M and CTX are as defined above, can be prepared by contacting cephalotaxin of formula H-O-CTX with a metal, amide, metal hydride or alkyl metal alone.

M may be an alkali metal such as lithium, potassium or sodium.

It is an object of the present invention to provide novel compounds such as:

- cephalotaxin lithium alkoxide corresponding to the following formula:

• ·

- cephalotaxin sodium alkoxide corresponding to the following formula:

a side chain cephalotaxan corresponding to the following formula and / or a salt thereof

R {CH

-CO-O-CTX wherein n is between 0 and 8;

Z is an oxygen, nitrogen or sulfur heteroatom;

R ⁵ , R ⁶ and R ⁸ are independently hydrogen;

a hydrocarbon group, saturated, unsaturated or aromatic, linear or branched and / or cyclic, in particular alkyl, alkenyl, alkynyl, cycloalkyl. cycloalkenyl, aryl, heterocycloalkyl, said group including or not including heteroatom (s);

an oxygenated ether bearing one of the above groups;

CTX is as defined above except for compounds wherein Z is an oxygen atom and

1 °) n = 2 or 3 and R ⁶ = R ⁸ = methyl and R ⁵ = OMe or hydroxyl, 2 °) n = 2 and R ⁶ = R ⁸ = methyl and R ⁵ = OMe or hydroxyl,

3 °) n = 3 and at the same time R ⁶ is hydroxyl when R ⁸ is methyl and R ⁵ is CH 2 CO 2 CH 3.

- side-chain cephalotaxan corresponding to the following general formula and / or a salt thereof:

R (CHJ

OH-CTX

2'mm, ^{R5, R6, R8} and CTX are as defined above;

except for the compound wherein m = 2, R ⁵ = CH 2 CO ² CH 3, R ⁶ = R ⁸ = methyl and CTX is as defined above.

R ⁵ is preferably -CH 2 CO-O-CH 3.

- side-chain cephalotaxan corresponding to the following general formula and / or a salt thereof:

CO-O-CTX wherein n, Z, Q ¹ , Q ² , R ⁵ , R ⁶ , R ⁸ , R ⁹ , R ¹⁰ and R ¹¹ are as defined above.

Preferably Q ² is an oxygen atom and / or Z is a nitrogen atom and the cephalotaxane has n = 0.

• ·

side-chain cephalotaxan corresponding to the following formula:

side-chain cephalotaxan corresponding to the following formula:

side-chain cephalotaxan corresponding to the following formula:

• · · ···

If the cyclic side chain of cephalotaxan with the side chain and / or a salt thereof is the following general formula:

R-CO-O-CTX wherein n, R ⁵ , R ⁶ , R ⁸ , CTX and Z are as defined above, said side chain is opened with a reagent and / or a protonic or non-protonic electrophilic radical E in an aqueous or non-aqueous medium to form an intermediate compounds of the following general formula:

OE

R-CO ₂ CTX wherein n, CTX, R ⁵ , R ⁶ and R ⁸ are as defined above, E is either hydrogen or a provisional or definite electrophile radical;

wherein the above intermediate compound may be attacked with a reagent or nucleophilic radical Z ', intentionally added or optionally present in the environment, and if the cyclic side chain of the side chain cefalotaxan and / or a salt thereof represents the following general formula:

wherein n, R ⁵ , R ⁶ , R ⁸ , R ⁹ , R ¹⁰ and R ¹¹ are as defined above and Z 1 is a heteroatom;

said side chain being opened by hydrolysis or cautious solvolysis in the presence of an activating agent or an opening aid.

Further, cephalotaxan with an open side chain of the following general formula is provided:

OF'

OH

RICH,

-CO ₂ CTX wherein n, CTX, R ⁵ , R ⁶ and R ⁸ are as defined above;

Z'je:

- either a halogen or a heteroatom bearing hydrogen or R ¹¹ as defined above;

or hydrogen, a hydrocarbon group wherein the radical bearing or not carrying heteroatom (s) is saturated, unsaturated or aromatic, linear or branched and / or cyclic, especially alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl or heterocycloalkyl.

For example, cephalotaxin esters of the following general formulas:

RR45

QH co ₂ ctx rQH

OH

-CH ₂ CO ₂ CTX

OH

-ÍCHJR ° CTX _{-CO2 CH2 CO2} Me

Me CH ₂ CO ₂ CTX CH ₂ CO ₂ Me wherein R ⁵ , R ⁶ , R ⁸ , Z ', X and CTX are as defined above;

- bromodeoxyharrintonin (n = 2) and bromodeoxyhomoharrintonin (n = 3)

Br

OH

CTX MeÍCHJMe _{-CO2 CH2 CO2} Me where CTX is as defined as above;

- aminodeoxyharrintonin (n = 2) and aminodeoxyhomoharrintonin (n = 3)

NH

OH

MeCO ₂ CTX

Me

CH ₂ CO ₂ Me wherein CTX is as defined above;

Further, if the cyclic side chain of cephalotaxan with the side chain and / or a salt thereof is the following general formula;

R <_{2> 2} ) _n -CO-O-CTX

Wherein n, R ⁵ , R ⁶ , R ⁸ , CTX and Z are as defined above, said chain is opened by treatment with a solution of hydrobromic acid in acetic acid in a halogenated solvent, preferably dichloromethane, followed by in sieve by hydrolysis, to give, without isolation of the intermediate, cephalotaxan with a side chain of the following general formula:

OH OH

R-CO ₂ CTX

R 'where n, CTX, R ⁵ , R ^e and R ⁸ are as defined above.

Acids corresponding to the following formula:

Ω-CO-OH wherein Ω is as defined above;

wherein said formula is equivalent to a racemic mixture comprising compounds of formulas (+) - Ω-ΟΟ-ΟΗ and (-) - Ω-ΟΟ-ΟΗ such that (+) - Ω-ΟΟ-ΟΗ represents the dextrorotatory enantiomer and (-) - Ω-ΟΟ-ΟΗ represents the levorotatory enantiomer, obtained

a) contacting said racemic mixture or one of its active forms of formula

CO-CO-A as defined above;

wherein said racemic mixture of said activated form individually generates:

- either the anion corresponding to formula (Ω-CO-O) ';

- or a cation corresponding to the formula (Ω-CO) *;

with the pure enantiomeric form of the chiral entity, said "cleavage agent" symbolized by Δ * (delta Stella) has the ability to form:

either a stable combination by covalent bonding;

or a convertible labile combination by hydrogen bonding or hydrophobic interaction;

or a labile intermediate by electrostatic interaction;

to give a diastereomeric mixture of Q-CO-O-Δ * and de Ω-CO-* *.

b) then physically separating a mixture of two diastereomers or two complex compounds or more generally two new entities physically and / or chemically different;

c) then regenerating and finally separating each enantiomer of the formula ΩCO-OH, where Ω * ("omega stel 1a") represents the general symbol of the same chiral radical in either one or other pure enantiomeric forms having the following formulas (+) - Ω-ΟΟ- ΟΗ and (-) - Ω-ΌΟ-ΟΗ, as defined above.

Preferably Ω-CO- is:

- either a group corresponding to the following general formula:

R (CH

2'n

-cokde Z, R ^6, R ⁸ and R ⁵ are as defined above;

- or a group corresponding to the following general formula:

wherein Z, R ⁶ , R ⁸ and R ⁵ are as defined above;

- or a group corresponding to the following general formula:

wherein n, R ⁵ , R ⁶ , R ⁸ , Z, Q ² , Q ¹ , R ⁹ , R ¹⁰ and R ¹¹ are as defined above.

The stable combination may be represented by an ester of the following formula Ω-CO-OA, where Ω and Δ * are as defined above, wherein the stable combination is obtained by contacting an acid with a chiral alcohol corresponding to the general formula ΗΟ-Δ *. of the process according to the invention.

The stable combination may be represented by an amide of the following formula Ω-COΝΗ-Δ, or Ω-ΟΟ-Ν-Δ * where Ω and Δ * are as defined above, wherein the stable combination is obtained by contacting the acid with a primary or secondary chiral amine corresponding to the general formula Η2Ν -Δ *, or ΝΝ = Δ *, where Δ * has the meaning given above according to the process of the invention.

The stable combination may be represented by a thioester of the following formula Ω-COS-Δ *, where Ω and Δ * are as defined above, wherein the stable combination is obtained by contacting an acid with a chiral thiol corresponding to the general formula HS-Δ *. according to the process of the invention.

Finally, the ionic combination may be represented by a salt prepared by contacting an acid with a chiral amine corresponding to one or more of the following three formulas:

Ω -CO-O '[ΝΗ-Δ *] ⁺

Ω -CO-O '[ΝΗ ₂ -Δ *] ⁺

Ω-CO-O '[ΝΗ ₃ -Δ *] ⁺ where Ω and Δ are as defined above.

An unstable combination can be prepared by chiral stationary phase chromatography.

Fractional crystallization initiated by a chiral precursor may produce a crystalline labile variant.

The chiral alcohol HO-Δ * is

- either (-) - quinine corresponding to the following formula:

- or (-) - or (+) - methyl imidate corresponding to the following formulas:

- or (-) - or (+) - menthol corresponding to the following formulas:

The chiral amine Η ₂ Ν-Δ is (-) - or (+) - ephedrine corresponding to the following formulas:

The present invention relates to the following novel compounds:

- (-) - quinidyl (2'R) - (-) - anhydrohomoharringtonate and (-) - quinidyl (2'S) - (-) anhydrohomoharringtonate corresponding to the following two formulas:

- (-) - menthyl (2'R) - (-) - anhydrohomoharringtonate and (-) - menthyl (2'S) - (-) anhydrohomoharringtonate corresponding to the following two formulas:

- (-) - methylmandelyl (2'R) - (-) - anhydrohomoharringtonate and (-) - methylmandelyl (2'S) (-) - anhydrohomoharringtonate corresponding to the following two formulas;

'' ·· 4 · · ♦ · · · · · ···

- (-) - ephedrine (2'R) - (-) - anhydrohomoharringtonate and (-) - ededrinium (2'S) - (-) anhydrohomoharringtonate corresponding to the following two formulas:

According to the process of the present invention, when the carboxylic acid is a tertiary heterocycloalkanoic carboxylic acid, corresponding to the following general formula:

R (CHJ

2> n

-CO-OH, wherein n, Z, R ^5, R ⁶ and R ⁸ are as defined above, said acid is obtained by treating in an aprotic solvent optionally in the presence of a cyclising agent ···· 4 44 4 •• ·· »t

444 · 4 4 · 4 · 4 4

And / or a dehydrating agent, said treatment being aided by the physical removal of the water produced.

- or an open tertiary ethylenic acid corresponding to the following general formula:

ZH o (CHJ

2'm where RN, Z, R ^5, R ⁶ and R ⁸ are as defined above.

- or an open tertiary ethylenic acid corresponding to the following general formula:

-.8

RZH

2'm

-CQ-OR wherein m is 1 to 8, Z, R ⁵ , R ⁶ and R ⁸ are as defined above, R ¹² is not a CTX group as defined above and is R ⁵ and / or an acid protecting group and / or a chiral group;

then R ^{12 is} removed later, either by saponification or hydrogenolysis, or more generally by a method known in the art to remove acid protecting groups.

In the absence of the cyclization additive, the cyclization reaction is carried out by heating.

Preferably, a protic acid such as sulfonic acid or formic acid or an aprotic acid included in the immobilized form is used as the cyclizing agent.

• ············ · 9 ·

In the acid preparation step described above, Z is an oxygen atom.

It is also an object of the present invention to prepare the following novel compounds:

- tertiary hydroxycycloalkanoic carboxylic acids including their salts and all racemic mixtures or various mixtures corresponding to the following general formula:

R ° (CH

2'n

-CO-OH wherein n, Z, R ⁵ , R ⁶ and R ⁸ are as defined above and R ⁵ is not hydrogen;

except for compounds where Z is an oxygen atom and,

°) n = 0 and R ⁵ is not -CH ₂ CO ₂ H or -CH ₂ CO ₂ CH 3;

2 °) n = 0 and R ⁵ is -CH ₂ CO ₂ H or -CH ₂ CO ₂ CH ₃ and R ⁶ = R ⁸ = methyl or -CH ₂ CO ₂ H or -CH ₂ CO ₂ CH 3;

3 °) n = 2 and simultaneously R ⁶ = R ⁸ = methyl and R ⁵ = OMe or hydroxyl;

4 °) n = 2 and at the same time R ⁶ = R ⁸ = methyl and R ⁵ is -CH ₂ CO ₂ H or CH ₂ CO ₂ CH ₃ or methyl;

5 °) n = 3 while R ⁶ is hydroxyl and R ⁸ is methyl and R ⁵ is CH ₂ CO ₂ CH ₃ ;

6 °) n = 3 and at the same time R ⁶ = R ⁸ = methyl and R ⁵ = OH or methyl or ethyl.

- tertiary oxacycloalkanoic carboxylic acid, including its salts and all its pure enantiomeric forms, or in racemic mixtures or various mixtures, corresponding to the following general formula:

R (CH

2'n

CO-O-H

R

Wherein n includes 0 to 8, R ⁵ , R ⁶ and R ⁸ are as defined above, but are not simultaneously hydrogen except for compounds corresponding to exceptions 1 to 6 as defined above.

- tertiary heterocycloalkanoic carboxylic acids, including their salts and all its pure enantiomeric forms or in racemic mixtures or various mixtures, corresponding to the following general formula:

R (CH

2'n

-CO-OR wherein n comprises 0 to 8, Z, R ⁵ , R ⁶ and R ⁸ are as defined above, R ⁵ is not hydrogen and R ¹² is not CTX as defined above;

except for compounds wherein Z is an oxygen atom and corresponding to exceptions 1 to 6 as defined above.

- a tertiary oxacycloalkane carboxylic hemiester, including its salts and all its pure enantiomeric forms, or in racemic mixtures or various mixtures, corresponding to the following general formula:

R - O (CH

2'n

-CO-O-H

CH ₂ CO ₂ Me wherein n comprises 0 to 8, R ⁶ and R ⁸ are as defined above, except for compounds corresponding to exceptions 1 to 6 as defined above.

- a tertiary oxacycloalkane carboxylic hemiester or anhydrohomoharringtonic acid, including their salts and all their pure enantiomeric forms or in racemic mixtures or various mixtures, corresponding to the following general formula:

R - O (CH

2'n

-co ₂ r ···· · ·· ··;

Wherein n comprises 0 to 8, R ⁶ and R ⁸ are as defined above, R ¹² is a hydrocarbon group other than CTX as defined above.

except for compounds corresponding to exceptions 1 to 6 as defined above.

- a tertiary oxacycloalkane carboxylic hemiester or anhydroharringtonic acid, including their salts and all their pure enantiomeric forms or in racemic mixtures or various mixtures, corresponding to the following formula:

Me

Me - O (CH

2'3

CO-O-H

CH ₂ CO ₂ Me

- a tertiary oxacycloalkane carboxylic hemiester or anhydroharringtonic acid, including their salts and all their pure enantiomeric forms or in racemic mixtures or various mixtures, corresponding to the following formula:

Me

Me (CH ₂ ) ₂ CO-OH

CH ₂ CO ₂ Me

- tertiary oxacycloalkanoic carboxylic acids, including their salts and all its pure enantiomeric forms or in racemic mixtures or various mixtures, corresponding to the following general formula:

Ph (CH) n-CO-OH wherein R ⁵ is as defined above, except for compounds corresponding to exceptions 1 to 6 as defined above.

- tertiary oxacycloalkanoic carboxylic acids, including their salts and all its pure enantiomeric forms or in racemic mixtures or various mixtures, corresponding to the following general formula:

Ph-O (CH

2'n

-CO-O-H wherein n is 1 to 8.

- tertiary oxacycloalkanoic carboxylic acids, including their salts and all its pure enantiomeric forms or in racemic mixtures or various mixtures, corresponding to the following formula:

Ph

R);

CO-O-H

CH ₂ CO ₂ Me where n is 0 to 8.

- tertiary oxacycloalkanoic carboxylic acid or oxahydro-neoharringtonic acid, including its salts and all its pure enantiomeric forms or in racemic mixtures or various mixtures, corresponding to the following formula:

- tertiary oxacycloalkanoic carboxylic acid or oxahydro-neoharringtonic acid, including its salts and all its pure enantiomeric forms or in racemic mixtures or various mixtures, corresponding to the following formula:

PhČH:

-CO-O-H

CH ₂ CO ₂ Me

- tertiary oxacycloalkanoic carboxylic acid, including its salts and all its pure enantiomeric forms or in racemic mixtures or various mixtures, corresponding to the following formula:

Ph-O

ČH;

-CO-O-H

- tertiary alkenoic carboxylic acids, including their salts and all its pure enantiomeric forms, or in racemic mixtures or various mixtures, corresponding to the following general formula:

* ·· e

· * * * * * * * * * * * · *

ΡΟΗ

2'm

-CO-O-H

R ⁶ wherein m includes between 1 and 8, R ⁶ and R ⁸ are as defined above, but are not simultaneously hydrogen and R ⁵ is not hydrogen or a heteroatom.

- tertiary alkenoic carboxylic acids, including their salts and all its pure enantiomeric forms, or in racemic mixtures or various mixtures, corresponding to the following general formula:

OH jCH)

2 ^of m

-CO-OH (CH ₂ CO _T R where m, R ^6, R ⁸ and R ¹² are as defined above am 'comprises between 1 and 8 respectively.

- tertiary alkenoic carboxylic acids, including their salts and all its pure enantiomeric forms, or in racemic mixtures or various mixtures, corresponding to the following general formula:

ΡΟΗ

2'm

-CO-O-H

Wherein m includes between 1 and 8, R ⁶ and R ⁸ are as defined above, but are not hydrogen.

- tertiary alkenoic carboxylic acids, including their salts and all its pure enantiomeric forms, or in racemic mixtures or various mixtures, corresponding to the following formula:

Me

Me — r

QH

-CO-O-H

CH ₂ CO ₂ Me

- tertiary alkenoic carboxylic acids, including their salts and all its pure enantiomeric forms, or in racemic mixtures or various mixtures, corresponding to the following formula:

Me

MeOH

CH;

-CO-O-H

CH ₂ CO ₂ Me

- tertiary alkenoic carboxylic acids, including their salts and all its pure enantiomeric forms, or in racemic mixtures or various mixtures, corresponding to the following formula:

\\ //

OH

ΊΟΗ ₂ )

2'm

-CO-O-H wherein m is 1 to 8, preferably m = 1.

- acid anhydrides of the general formula Ω -CO-O-CO-Ω wherein Ω is as defined above.

mixed acid anhydrides of the formula Ω-CO-A, wherein A is a group selected from the following substituents:

methoxyformyloxy of formula MeOCOO-, trifluoroacetyloxy of formula CF3COO-, alkylsulfonyloxy of formula RSO3-, phosphoxy of formula (RO) 2POhalophosphoxy of formula ROP (CI) Otrialkylsilyloxy of formula R ₃ SiO-, of the formulas where R is alkyl, dimethylformamidium chloride of formula o

Me

Cl

Me acyloxypyridinium bromide of formula

and 2,4,6-trichlorobenzyloxy.

- mixed anhydrides corresponding to the following formula:

- acid chlorides as defined above, corresponding to the general formula Ω-CO-X, where X is halogen.

- cyclic anhydrides corresponding to the following general formula:

· · · · · · · · · · · · · · · · · · · · · · · · · · ·

wherein n, R ⁶ and R ⁸ are as defined above.

- cyclic anhydrides corresponding to the following formula:

In the process of the invention, the side chain cephalotaxan is purified as a salt by chromatography using a hydrophobic reverse phase as a stationary phase and a mobile phase without an organic solvent such as a solution adjusted to pH 2-4 of a buffer prepared with acids and bases or ammonium salt and one or a plurality of additives as a silanol buffer, wherein said cephalotaxan salt is generated from a mineral acid in the form of a chlorohydrate, sulfate, phosphate, perchlorate or organic acid in the form of a tartrate, maleate, citrate or lactate.

In the process according to the invention, the side-chain cephalotaxan is purified by chromatographic purification of natural or semisynthetic or synthetic homoharringtonine for pharmaceutical use and corresponding to the following formula:

in order to remove undesirable related impurities, in particular 2'-epihomoharringtonine, arising from:

(a) either from a semi-synthetic procedure using synthetic homoharringtonic acid of inadequate enantiomeric purity, the impurity generated having an absolute configuration corresponding to the following formula:

(b) from a biosynthetic process in plants using cephalotaxin of inadequate enantiomeric purity or in the form of an artifact, by partial racemization of the cephalotaxin moiety, wherein the impurity generated exhibits strictly identical chromatographic properties with a non-chiral system, with an absolute configuration opposite to the above (enantiomer) the following formula:

in particular using one of the following chromatographic systems:

A) Stationary phase: alkyl- or phenyl- or alkylphenyl- or phenylalkylsilane, preferably n-octadecylsilane,

B) Mobile phase: water-tetrahydrofuran, water-methanol, water-acetonitrile or buffer pH 2 to 6.5 to replace water or another mobile phase with equivalent selectivity.

The purification procedure and chromatographic control of natural or semi-synthetic or synthetic homoharringtonine allows to compensate for the double deficiency in the enantiomeric purity of the semi-precursors, both the side chain precursors (said homoharringtonic acid) and cephalotaxin, both precursors independently generated by the overall synthetic or semi-synthetic process. (biosynthesis) and, in fact, removal of the unnatural enantiomer of homoharringtonine having the opposite absolute configuration, using a chiral stationary phase on a production scale.

The present invention is illustrated, but not limited, by the following schemes.

Scheme 1 in the Annex provides a definition of the main harringtonines.

Scheme 2 summarizes the prior art harringtonine preparation process. Scheme 3 provides a homoharringtonine preparation sequence corresponding to Example 25, wherein A is a 2,4,6-trichlorophenyl group, R is methyl, and R 'is a cephalotaxyl moiety.

Scheme 4 represents a variant of the process of the invention, more specifically the semisynthesis of haringtonines via oxacycloalkanoic carboxylic acids. The substituents R ⁶ , R ⁷ , R ⁸ , R, A, CTX and the letters n and m in this scheme are defined in the description.

The following definitions apply throughout the document.

Alkaloids: natural substances present in the plant kingdom with at least one cyclic or acyclic basic nitrogen (possible extension: animal kingdom, primary amine; impossible extension: amidic nitrogen, since it is not basic, for example taxanes) and often exhibiting pronounced pharmaceutical properties.

Basic or free alkaloid: an alkaloid with a tertiary amine in unionized form, usually existing at alkaline pH and soluble in aprotic organic solvents.

• ·

Alkaloid salt or just "salt": an ionized form of an amine-function alkaloid having a positive formal charge and a negative counterion, in fact more soluble in water and protic solvents and less soluble in aprotic solvents.

Cephalotaxanes 1 (see Scheme 1): this generic term indicates a basic system having differently oxygenated substituents (aliphatic or aromatic ether, free or esterified alcohol, enol and / or free or substituted phenol, bridging ether, more generally all substituents usually found in natural state for this kind of compounds).

Cephalotaxins 2: this generic term indicates cephalotaxanes with a possible at least one substituent listed above, the side chain being excluded.

Cephalotaxin 2: cephalotaxin, which is mainly found in the genus Cephalotaxus.

Cephalotaxoids: this generic term indicates cephalotaxins that are of unnatural origin.

Harringtonines: this generic term indicates cephalotaxan with at least one alcohol group, phenol or enol, esterified with a side chain and optionally one or more substituents described above.

Harringtonine (the): one of the major alkaloids carrying the side chain at position

3.

Harringtoids: this generic term indicates harringtonine of unnatural origin, where the side chain is an ester group with at least three carbon atoms.

Harringtoid side chain: this is a generic term for an ester formed between one hydroxyl group and a carboxylic acid containing at least three carbon atoms, usually with a tertiary alcohol group at position a and a hydrophobic substituent at position ω relative to the carboxyl group.

Adaptation of additive and elimination empirical nomenclature for cephalotaxans.

• 4

Prefixes commonly used in the literature are used to determine structural variations of the harringtonine side chain (see examples in the table of Scheme 1). The side chain of the reference is the harringtonines shown in Formula 3b, n = 2, R = H, R ⁷ = H, R ⁶ = 2-hydroxy isopropyl.

Homo: 1 extra carbon atom.

Bishomo: 2 extra carbon atoms.

Nor: side chain containing 1 carbon atom less.

Iso: side chain with methylene bearing a hydroxyl group at the juxtaterminal carbon site.

Deoxy: the hydroxyl group of the juxtaterminal carbon is replaced by hydrogen.

Anhydro: two tertiary hydroxyl groups lose a water molecule to yield a saturated oxygen heterocycle.

neo: R ⁶ is a phenyl group in place of 2-hydroxypropyl.

HPLC: high performance liquid chromatography.

NMR: nuclear magnetic resonance.

• ·

The following examples are provided to further illustrate the invention, and are not to be construed as limiting the scope of the invention in any way.

DETAILED DESCRIPTION OF THE INVENTION

Example 1

Preparation of ethyl 2-methoxycarbonylmethyl-2-hydroxy-6-methylhept-5-enoate or ethyl 6-desoxy-5,5-dehydrohomoharringtonate

CO ₂ Et

CH ₂ CO ₂ Me

°) Preparation of intermediate oxalate

5-Bromo-2-methyl-pent-2-ene (15.6 g, 95.6 mmol) was added dropwise to a stirred mixture of magnesium (2.32 g, 95.5 mmol) (activated with iodine crystal) in anhydrous tetrahydrofuran (75 mL). The onset of the reaction is accompanied by vigorous heating and refluxing of the reaction mixture. Reflux is maintained until most of the magnesium has reacted and the reaction mixture is then diluted with anhydrous tetrahydrofuran (150 mL). To a stirred mixture of diethyl oxalate (10.8 mL, 80 mmol) in anhydrous tetrahydrofuran (75 mL) was added Grignard's reagent at -75 ° C over 20 minutes. Stirring is maintained at -78 ° C ± 5 ° C for 30 minutes and then the temperature is raised to -10 ° C over 1.5 hours. The mixture was cooled with a 15% ammonium chloride solution (300 mL). The separated organic layer was washed with 15% ammonium chloride solution (300 mL) and evaporated to dryness. The aqueous layer was extracted with ether (2 x 300 mL). The organic layers were combined with the concentrate and washed with brine (300 mL), dried over magnesium sulfate and evaporated to dryness. The crude product was purified by distillation to give a colorless oil (10.3 g, 70%). The intermediate α-ketoester has the following characteristics:

Φ φ φ φ φ · · · · · · · · • · · · · · · · · · · · · · φ φ φ

IR (ATR) (cm ^-1 ): 2790; 2916; 1725; 1068.

¹ H NMR 400 MHz (CDCl ₃ ) (δ ppm, J Hz):

5.08 (1H, J, J = 7.2, HC =); 4.32 (2Η, q, J = 7.1, OCH ₂ ); 2.86 (2H, t, J = 7.2, _CH2 CO);

2.32 (2H, q, J = 7.2, _CH2 -C =); 1.68 (3H, s, CH ₃ ); 1.62 (3H, s, CH ₃ ); 1.37 (3 H, t, J =

7.1, OCH ₂ CH ₃ ).

2 °) Preparation of the title product

Anhydrous methyl acetate (0.6 mL, 7.5 mmol) was added at -78 ° C over 1 minute to a stirred commercial 1M solution of lithium bis- (trimethylsilylamide) in tetrahydrofuran (7.5 mL, 7.5 mmol) and the reaction Allow to proceed at -78 ± 5 ° C for 20 minutes. To a stirred mixture of ethyl 2-oxo-6-methylpent-5-enoate prepared above (480 mg, 2.6 mmol) in anhydrous tetrahydrofuran (10 mL) at 78 ° C was added lithium enolate over 5 minutes and the resulting mixture was stirred at -78 ± 5 ° C for 30 minutes. After monitoring the CCM, the freezing bath was removed and the mixture was cooled with 15% ammonium chloride solution (10 mL). The separated organic layer was washed with 15% ammonium chloride solution (10 mL) and evaporated to dryness. The aqueous layers were extracted with ether (2 x 10 mL). The organic layers were combined with the concentrate and washed with brine (10 mL), dried over magnesium sulfate and evaporated to dryness. The resulting crude product (1.13 g) was purified by column chromatography (cyclohexane / ethyl acetate (95: 5), silica (15-40 µm) 38 g) to give a colorless oil (482 mg, 72%). the product thus obtained exhibits the following characteristics:

IR (ART) (cm ^-1 ): 3508; 2969; 2919; 1732; 1438; 1193.

¹ H NMR 400 MHz (CDCl ₃ ) (δ ppm, J Hz):

5.05 (1H, t, J = 7.1, HC =); 4.27 (2H, q, J = 7.1, _OCH2); 3.70 (1 H, s, OH); 3.68 (3H, s,

OCH ₃ ); 2.92 and 2.70 (2H, 2d, J _? = 16.1. CH _? CO?); 2.12 (1 H, m); 1.88 (1 H, m); 1.72 (2 H, m); 1.67 (3H, s, CH ₃ ); 1.58 (3H, s, CH ₃ ); 1.31 (3H, t, J = 7.1, OCH ₂ CH ₂ CH ₃ ).

Example 2 · »·· • ··

Preparation of ethyl 2-methoxycarbonylmethyl-2-hydroxy-6,6-dimethyl-2-tetrahydropyran carboxylate or ethyl anhydrohomoharrinetonate

1 °) Method A p-Toluenesulfonic acid (2.06 g, 10.8 mmol) was added to a stirred solution of the ethylene ester resulting from Example 1 (2.8 g, 10.8 mmol) in toluene (30 mL) and the resulting the mixture was stirred at 65 ° C for 5 hours. After cooling to room temperature, the mixture was hydrolyzed with saturated sodium bicarbonate solution. The aqueous layer was extracted with ether (3 x 50 mL) and the organic layers were combined, washed with brine (100 mL), dried over magnesium sulfate and evaporated to dryness. The resulting crude product (2.8 g) was purified by column chromatography (cyclohexane / ether (95: 5), silica (15-40 µm) 110 g) to give a colorless oil (1.94 g, 69%). The product thus obtained exhibits the following characteristics:

¹ H NMR 400 MHz (CDCl ₃ ) (δ ppm, J Hz):

4.21 (2H, m, OCH 2 CH 3); 3.64 (3H, s, _OCH3); 2.85 and 2.60 (2H, d, J _AB = 14.0, CH 2 CO 2); 2.30 (1H, dt, J = 13.3 and 3.7); 1.87 (1H, qt, J = 13.8 and 3.6); 1.62 (1 H, m);

1.51 (2 H, m); 1.43 (1 H, m); 1.31 (3H, t, J = 7.1, OCH 2 CH 3); 1.22 (3H, s, CH _3); 1.13 (3H, s, _CH3).

2 °) Method B

To a stirred solution of the ethylene ester formed in Example 1 (50 mg, 0.19 mmol) in methanol (30 mL) was added 1N hydrochloric acid (0.5 mL) and the resulting mixture was stirred at 65 ° C for 15 hours. After dilution with dichloromethane, the organic layer was dried over magnesium sulfate and evaporated to dryness. The resulting crude product (32 mg) was purified by column chromatography (dichloromethane, then dichloromethane / methanol (9: 1), silica (15-40 µm), 2.2 g) to give • 9999 9 ·· · 9 99 • · · 9 · · 9 ····

9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 999 9999 99 99 expected intermediate diol (20 mg, 37%). The product thus obtained exhibits the following characteristics:

Me (CH ₂ ) ₃ CH ₂ CO ₂ Me

IR (ATR) (cm ^-1 ): 3490; 2966; 1731; 1193; 1177; 1152.

¹ H NMR 400 MHz (CDCl ₃ ) (δ ppm, J Hz):

4.28 (2H, q, J = 7.2, _OCH2); 3.75 (1 H, s, OH); 3.68 (3H, s, _OCH3); 2.93 and 2.69 (2H, 2d, J _AB = 16.2, CH 2 CO 2); 1.70 (2 H, m); 1.53 (1 H, m); 1.44 (1 H, m); 1.30 (3 H, t, J =

7.1, OCH ₂ CH ₃ ); 1.20 (3H, s, CH _3); 1.19 (3H, s, CH ₃ ).

To a stirred solution of the diol prepared above (19 mg, 0.069 mmol) in 1,2-dichloroethane (1.4 mL) was added anhydrous zinc chloride (10 mg, 0.069 mmol) and the resulting mixture was stirred at 80 ° C for 1.5 h. . After cooling to room temperature, the mixture was washed with water, then brine, and the aqueous layer was extracted three times with dichloromethane. The combined organic layers were dried over magnesium sulfate and evaporated to dryness to give the cyclic diester (7 mg, 40%). The product thus obtained exhibits identical characteristics to those of the product obtained by Method A.

3 °) Method C

A solution of the ethylene ester formed in Example 1 (400 mg, 1.55 mmol) in a mixture of formic acid (4 mL) and water (4 mL) was stirred at 50 ° C for 15 hours. After removal of the formic acid in vacuo, the residue was treated with 5% sodium bicarbonate solution. The aqueous layer was extracted three times with dichloromethane, then the combined organic layers were dried over magnesium sulfate and evaporated to dryness. The resulting crude product (375 mg) was purified by column chromatography (dichloromethane, then dichloromethane / methanol (98: 2), silica (15-40 µm) 16 g) to give a colorless oil (235 mg, 55%). The product thus obtained exhibits the same characteristics as the product obtained by Method A. Cyclization of the diol thus obtained. With zinc chloride, similar to Example 2, Method B above, results in cyclic diesters which exhibit identical characteristics to the product obtained in Method A.

Example 3

Preparation of 2-carboxymethyl-2-hydroxy-6-methylhept-5-enoic acid or Odemethyl-6-desoxy-5,6-dehydrohomoharringtonic acid:

CO, H ch ₂ every ₂ h

A mixture of potassium hydroxide (14.2 g, 252 mmol) in water (170 mL) was added to a stirred solution of the ethylene ester formed in Example 1 (10.95 g, 42 mmol) in methanol (300 mL) and the resulting mixture was stirred at reflux for 1.5 hours. After cooling to room temperature and removing the methanol in vacuo, the residue was treated with water (10 mL) and the resulting aqueous layer was extracted with ether (250 mL). After acidification (pH 1) with 10% hydrochloric acid, the aqueous layer was extracted with ether (3 x 250 mL). The combined organic layers were dried over magnesium sulfate and evaporated to dryness to give a solid (8.66 g, 95%). The product thus obtained exhibits the following characteristics:

IR (ATR) (cm ^-1 ): 3500; 3019; 2966; 2931; 1716; 1691; 1656; 1219; 1199; 1111.

¹ H NMR 400 MHz (CDCl ₃ ) (δ ppm, J Hz):

5.06 (1H, t, J = 6.9, HC =); 3.04 and 2.78 (2H, 2d, J _AB = 17.1, _CH? CO); 2.25 to 1.20 (4H, m, 2 x CH _2); 1.67 (3H, s, CH ₃ ); 1.60 (3H, s, _CH3).

Example 4 • ·

Preparation of 2-carboxymethyl-6,6-dimethyl-2-tetrahydro-pyrancarboxylic acid or O-demethylanhydrohomoharrine acid:

°) Method A

A mixture of potassium hydroxide (4.2 g, 75 mmol) in water (45 mL) was added to a stirred solution of the cyclic diester formed in Example 2 (1.94 g, 7.5 mmol) in ethanol (75 mL). The mixture was stirred at reflux for 5 hours. After cooling to room temperature and removing the ethanol in vacuo, the residue was treated with water (10 mL) and the resulting aqueous layer was extracted with ether (2 x 50 mL). After acidification with 2N hydrochloric acid (35 mL), the aqueous layer was saturated with sodium chloride and then extracted with ether (3 x 50 mL). The combined organic layers were washed with brine (2 x 100 mL), dried over magnesium sulfate and evaporated to dryness to give a pale yellow oil (1.66 g, 98%). The product thus obtained exhibits the following characteristics:

IR (ATR) (cm ^-1 ): 2974; 2941; 1709; 1215.

¹ H NMR 400 MHz (CDCl ₃ ) (δ ppm, J Hz):

3.01 and 2.95 (2H, 2d, J _AB = 1.16 _{CH? CO?);} 1.89 (1 H, m); 1.75 (2 H, m, CH); 1.58 (3 H, m); 1.31 (6H, s, 2 x _CH3).

2 °) Method B

To a stirred solution of the ethylene dibasic acid formed in Example 3 (50 mg, 23 mmol) in anhydrous toluene (500 μΙ) was added zinc chloride (6 mg, 0.04 mmol) and the resulting mixture was stirred at 80 ° C for 15 hours . After cooling to room temperature, the mixture was hydrolyzed with 10% hydrochloric acid and the resulting aqueous layer was extracted three times with ethyl acetate. The combined organic layers were dried over magnesium sulfate and evaporated to dryness to give a pale yellow solid (38 mg, 76%). The product thus obtained exhibits identical characteristics to the product obtained by Method A.

• ·

Method C

A solution of the ethylene dibasic acid formed in Example 3 (50 mg, 0.23 mmol) in a mixture of formic acid (500 µL) and water (500 µL) was stirred at 60 ° C for 3 hours. After cooling to room temperature and removal of formic acid in vacuo, the residue was treated with ethyl acetate. The resulting organic layer was washed with 10% hydrochloric acid and the aqueous layer was extracted three times with ethyl acetate. The combined organic layers were dried over magnesium sulfate and evaporated to dryness to give a pale yellow solid (50 mg, 100%). The product thus obtained exhibits the same characteristics as the product obtained in Method A.

Example 5

Preparation of 2-methoxycarbonylmethyl-2-hydroxy-6-methylhept-5-enoic acid and 6-desoxy-5,6-dehydrohomoharringtonic acid:

A mixture of the ethylene dibasic acid of Example 3 (500 mg, 2.3 mmol) and a commercial solution of boron trifluoride-methanol complex in methanol (4.5 mL, BF ₃ 12% w / w) was stirred at 18 ± 5 °. C for 16 hours. After carefully adding the reaction mixture to saturated sodium bicarbonate solution (50 mL), the resulting aqueous layer was washed with ether (50 mL), acidified (pH 1) with 2N hydrochloric acid (0.5 mL) and extracted with ether (3 x 50 mL). . The combined organic layers were dried over magnesium sulfate and evaporated to dryness to give a viscous yellow oil (310 mg, 58%). The crude product thus obtained exhibits the following characteristics:

IR (ATR) (cm ^-1 ): 3483; 2954; 1731; 1197; 1173.

¹ H NMR 400 MHz (CDCl 3) (δ ppm, J Hz):

5.06 (1 H, m, HC =); 4.12 (2H, br. S, CO ₂ H + OH); 3.73 (3H, s, _OCH3); 2.99 and 2.74 (2H,

2d, Jab = 16.7, CH 2 CO 9): 2.16 (1H, m); 1.98 (1 H, m); 1.85 - 1.60 (4H, m); 1.67 (3H, s, _CH3); 1.60 (3H, s, _CH3).

Example 6

Preparation of 2-methoxycarbonylmethyl-6,6-dimethyl-2-tetrahydropyranecarboxylic acid or anhydrohomoharrine acid:

1 °) Preparation from dibasic acid

A mixture of the cyclic dibasic acid formed in Example 4 (1.6 mg, 7.4 mmol) and a commercial solution of boron trifluoride-methanol complex in methanol (15.5 mL, BF ₃ 12% w / w) was stirred at 18 ± 5 ° C for 15 hours After carefully adding the reaction mixture to saturated sodium bicarbonate solution (50 mL), the resulting aqueous layer was washed with ether (2 x 50 mL) (see addition preparation below), acidified (pH 1) with 2N hydrochloric acid (15 mL) and extracted with ether (3 x 75 mL) The combined organic layers were dried over magnesium sulfate and evaporated to dryness to give a yellow oil (1.17 g, 69%). :

IR (ATR) (cm ^-1 ): 2974; 2951; 1740; 1718; 1437 ¹ H NMR 400 MHz (CDCl ₃ ) (δ ppm, J Hz):

3.70 (3H, s, _OCH3); 3.03 and 2.98 (2H, 2d, J _AB = 16.1, CH7CO7); 1.82 (1 H, m); 1.74 (3 H, m); 1.62 (1 H, m); 1.48 (1 H, m); 1.31 (3H, s, CH _3); 1.26 (3H, s, _CH3).

Additional Preparations • · · · · · · · · · · ·

a) Obtaining a diester

Me (CH ₂ ) ₃ Cl 2 CO / Zle

The combined organic layers mentioned above were dried over magnesium sulfate and evaporated to dryness to give a mixture of diester and monoesters (396 mg). After working up this mixture with saturated sodium bicarbonate solution, the aqueous layer was extracted with ether and the resulting organic layer was dried over magnesium sulfate and evaporated to dryness to give an oil (292 mg, 17%). The product thus obtained has the following characteristics:

¹ H NMR 400 MHz (CDCl ₃ ) (δ ppm, J Hz):

3.75 (3H, s, _OCH3); 3.65 (3H, s, _OCH3); 2.85 and 2.61 (2H, 2d, J _AB = 14.1, CH 7 CO 7);

1.85 (1 H, m); 1.62 (1 H, m); 1.50 (2 H, m); 1.43 (1 H, m); 1.21 (3H, s, CH _3); 1.11 (3H, s,

b) Obtaining the regio-hemiester by mono-saponification of the above diester

Me O CO, Me

Me (CC ₂ ) ₃ CH ₂ CO ₂ H

To a stirred solution of the above cyclic diester (285 mg, 1.17 mmol) in methanol (11 mL) was added a mixture of potassium hydroxide (654 mg, 11.7 mmol) in water (7 mL) and the resulting mixture was stirred at room temperature. 30 minutes. After removal of the methanol in vacuo, the residue was treated with water (7 mL) and the resulting aqueous layer was extracted three times with ether. After acidification (pH 1) with a 10% hydrochloric acid solution, the aqueous layer was extracted. The combined organic layers were dried over magnesium sulfate and evaporated to dryness. The resulting crude product (236 mg) was purified by column chromatography (dichloromethane / methanol (95: 5), silica (15-40)). 6.5 g) to give a pale yellow solid (220 mg, 82%). The product thus obtained exhibits the following characteristics:

IR (KBr) (cm ^-1 ): 3421; 2960; 2929; 1744; 1705; 1209.

¹ H NMR 400 MHz (CDCl ₃ ) (δ ppm, J Hz):

3.76 (3H, s, _OCH3); 2.76 and 2.67 (2H, 2d, J _AB = 15.3, CH? _CO?): 2.36 (1H, m, J _AB =

13.7, J ₃ -4-3.5, J ₃ -5-1.2, H-3eq); 1.85 (1H, m, J _AB -Jax-ax ⁼ 14.0, Jax-oq ⁼ 3.7, H-4ax);

1.67 (1H, m, J _AB = 14.1, _{J-3, 5} = 3.9, H-4 _eq); 1.59 (1H, m, J _AB = 13.4, J = 3.6, J ₅ = ₃ )

1.0, H-5 ( _q ); 1.49 (1H, m, J _AB -Jax-ax-13.2, Jax-oq ⁼ 4.0, H-3ax); 1.42 (1H, m, J _AB -Jaxax = 13.2, Jax-oq-4.5, H-5ax); 1.33 (3H, s, CH ₃ ); 1.16 (3H, s, _CH3).

2 °) Preparation from ethylene hemiester

To a stirred solution of the ethylene hemiester ester formed in Example 5 (4.6 g, 20 mmol) in toluene (125 mL) was added p-toluenesulfonic acid (3.8 g, 20 mmol) and the resulting mixture was stirred at 65 ° C for for 5 hours. After cooling to room temperature, the mixture was hydrolyzed with saturated sodium bicarbonate solution (100 mL). The aqueous layer was washed with ether (2 x 100 mL) and the organic layers were removed (to eliminate the diester formed). After acidification (pH 1) with 1N hydrochloric acid (35 mL), the aqueous layer was saturated with sodium chloride and then extracted with ether (3 x 100 mL). The combined organic layers were washed with brine (100 mL), dried over magnesium sulfate and evaporated to dryness. The resulting crude product (3.9 g) was purified by column chromatography (dichloromethane / methanol (99: 1), silica (15-40 µm) 160 g) to give a yellow oil (3.1 g, 67%). The crude product thus obtained exhibits the same characteristics as the product obtained above.

Example 7

Preparation of 2-carboxymethyl-2-hydroxy-6-methylhept-5-enoic acid cyclic anhydrides

Me O ll

Me (CH ₂ ) ₃ ··················· · ··· · · ··· «·· ······· · · ··

A mixture of the cyclic dibasic acid formed in Example 4 (245 mg, 1.1 mmol) and acetic anhydride (4 mL) was stirred at reflux for 16 hours. After evaporation of the reaction mixture in vacuo, the residue was treated with toluene and evaporated again in vacuo to give a viscous yellow oil (189 mg, 84%). The product thus obtained exhibits the following characteristics:

IR (ATR) (cm ^-1 ): 2976; 2951; 1732; 1188; 1170.

¹ H NMR 400 MHz (CDCl ₃ ) (δ ppm, J Hz):

3.02 (2H, s, CH 2 CO 2); 1.98 (2H, m, _CH2); 1.8-1.5 (4H, m, CH _2); 1.31 (3H, s, CH _3);

1.22 (3H, s, _CH3).

Example 8

Preparation of (-) - cephalotaxyl pivalate:

1 °) Mixed anhydride method

To a stirred mixture of pivalic acid (100 mg, 0.98 mmol) in anhydrous toluene (2 mL) at room temperature was added triethylamine (dried over potassium hydroxide) (138 µL, 0.98 mmol) and 2,4,6- trichlorobenzoyl chloride (153 μΙ, 0.98 mmol). The mixture was stirred at 18 ± 5 ° C for 1.5 hours (the disappearing starting acid was controlled by IR), 4-dimethylaminopyridine (139 mg, 1.14 mmol) was added and the reaction was allowed to react for 5 minutes and cephalotaxin (103) was added. mg, 0.33 mmol). The mixture was stirred at 18 ± 5 ° C for 15 minutes, the reaction mixture was filtered through filter paper and diluted with ether (5 mL). The resulting organic layer was washed successively with water (5 mL), saturated sodium bicarbonate solution (5 mL), again water (5 mL), and then dried over sulfate. · Magnesium and evaporate in vacuo. The resulting crude product was purified by column chromatography (dichloromethane / methanol (98: 2), silica (15 to 40 μηη)) to give a solid (130 mg, 93%).

¹ H NMR 400 MHz (CDCl ₃ ) (δ ppm, J Hz):

6.60 (1 H, s, H-17); 6.58 (1 H, s, H-14); 5.84 and 5.83 (2H, d, J _AB = 1.5, OCH ₂ O); 5.83 (1H, d, H-3); 5.02 (1 H, s, H-1); 3.77 (1H, d, = 9.6, H-4); 3.69 (3H, s, _OCH3), 3.21 (1H, m, _JAB = 14.0, J = 12.5, 7.8, H-11b); 3.09 (1 H, m, H-8a); 2.94 (1 H, td, J = 11.5,

7.1 H-10a); 2.57 (2H, m, H-8b + H-10b), 2.35 (1H, dd, J _AB = 14.5, J = 6.9, H-11a); 2.03 (1H, td, J _AB = 12.1, J = 9.7, H-6 _A ); 1.89 (1H, m, J _AB = 12.1, J = 7.9, 4.0, H-6 _A );

1.75 (2H, m, _CH2 -7); 0.83 (9H, s, C (CH _{3) 3).}

2 °) Method using DCC

To a stirred mixture of pivalic acid (50 mg, 0.49 mmol) in anhydrous toluene (2 mL) maintained under an inert atmosphere was added 1,3-dicyclohexylcarbodiimide (130 mg, 0.63 mmol). The mixture was stirred at room temperature for 10 minutes, and cephalotaxin (50 mg, 0.16 mmol) and pyrrolidine pyridine (24 mg, 0.16 mmol) were added. Stir the mixture at 18 ± 5 ° C for 2 hours and then at 50 ° C for 15 hours (reaction is controlled by CCM, eluent dichloromethane / methanol 9: 1), filter the reaction mixture on a ground glass filter, wash the cake toluene (5 mL) and the filtrate evaporated in vacuo. The resulting crude product (130 mg) was purified by column chromatography (dichloromethane / methanol (9: 1), silica (15-40 µm) 3 g) to give a white solid (36 mg, 57%). The crude product thus obtained shows the same characteristics as the product obtained through the mixed anhydride.

Example 9

Preparation of (-) - cephalotaxyl 2-methoxycarbonylmethyl-6,6-dimethyl-2-tetrahydro-pyrancarboxylate or anhydrohomoharringtonine and methyl 2cephalotaxyloxycarbonylmethyl-6,6-dimethyl-2-tetrahydropyrancarboxylate from the cyclic anhydride formed from Example 7:

To a stirred mixture of the anhydride formed in Example 7 (50 mg, 0.24 mmol) in anhydrous dichloromethane (0.5 mL) at room temperature was gradually added pyridine (250 μΙ, 3.1 mmol), pyrrolidinopyridine (10 mg, 0, 07 mmol) and cephalotaxin (76.4 mg, 0.24 mmol). The mixture was stirred at 18 ± 5 ° C for 48 hours and then added gradually

1,3-dicyclohexylcarbodiimide (100 mg, 0.48 mmol), methanol (60 μΙ, 1.5 mmol), pyrrolidinopyridine (10 mg, 0.07 mmol) and toluene (1 mL). The mixture was stirred at 18 ± 5 ° C for 24 hours (reaction monitored by CCM), the reaction mixture was filtered and the filtrate was evaporated in vacuo. The resulting crude product was purified by column chromatography (dichloromethane / methanol (99: 1), silica (15-40 µm)) to give the expected product (12 mg, two diastereomers) contaminated with regioisomer * (two diastereomers) resulting from anhydride opening. The expected product thus obtained has the following characteristics:

¹ H NMR 400 MHz (CDCl ₃ ) (δ ppm, J Hz):

6.61 (1 H, s, H-17); 6.57 (1 H, s, H-14 *); 5.91 (J 3-4 = 9.8) and 5.84 (2H, 2d, H-3); 5.84 and 5.79 (2d, J _A b = 1.4, OCH ₂ O); 5.84 and 5.82 (2d, J _AB = 1.4, OCH ₂ O); 5.04 and 5.01 (1H, 2s, H-1); 3.79 and 3.78 (1H, 2d, J 4-3 = 9.6, H-4); 3.70 and 3.65 (3H, 2s, _OCH3); 3.59 (3H, s, _OCH3); 3.15 (1H, m, J-11β); 3.09 (1H, m, H-8a); 2.94 (1 H, m, H-10a); 2.58 (2H, m, Η-8β + Η-10β); 2.37 (1 H, m, H-11a); 2.16 and 1.81 (2d, J _AB = 14.4, CH ₂ CO ₂ ); 2,13 a

1.66 (2d, J _AB = 14.3, CH ₂ CQ _2); 2.02 (1 H, m, H-6 _A ); 1.88 (1 H, m, H-6 _B ); 1.75 (2H, m, _CH2 -7); 1.8 to 1.2 (6H, m, 3 x _CH2); 1.11 and 1.02 (2s, 2 x CH ₃ ); 1.10 and 1.04 (2s, 2 x CH ₃ ).

‘The regioisomer listed above is also obtained under the following conditions:

····· · · ···· · · ···· · · · ···· · · · · ♦ ## ··· · · · · · · To a stirred mixture of the hemiester resulting from Example 6, Method C (100 mg, 0.43 mmol) in anhydrous toluene (1 mL) maintained under an inert atmosphere at 1,3-Dicyclohexylcarbodiimide (120 mg, 0.58 mmol) was added at room temperature. The mixture was stirred for 5 minutes and cephalotaxin (45 mg, 0.15 mmol) and pyrrolidine pyridine (21 mg, 0.14 mmol) were added. The mixture was stirred at 35 ° C for 45 minutes and then at 8 ° C for 15 minutes (CCM reaction control, eluent dichloromethane / methanol, 9: 1), the reaction mixture was filtered, the cake washed with toluene (5 mL) and the filtrate was evaporated in vacuo. The resulting crude product was purified by column chromatography (dichloromethane / methanol (98: 2), silica (15-40 µm) 4 g) to give the expected product (23 mg, 30%, two diastereomers). The product thus obtained exhibits the following characteristics:

¹ H NMR 400 MHz (CDCl ₃ ) (δ ppm, J Hz):

6.61 and 6.58 (1H, 2s, H-17 *); 6.57 and 6.53 (1H, 2s, H-14 *); 5.89 and 5.86 (2d, J AB = 1.5, OCH ₂ O); 5.87 and 5.85 (2d, _JAB = 1.5, _OCH2 O); 5.76 (1H, d, J _3-4 = 9.4, H-3); 5.02 (1 H,

2s, H-1); 3.73 and 3.72 (1H, 2d, J 4-3 = 9.4, H-4); 3.70 and 3.68 3H, 2s, _OCH3); 3.69 and 3.65 (3H, 2s, _OCH3); 3.15 (1 H, m, H-11a); 3.07 (1H, m, H-8a); 2.90 (1H, m, H-10a); 2.74 and 1.95 (2d, J _AB = 15.3, CH ₂ CO ₂ ); 2.56 (2H, m, Η-8β + Η-10β); 2.33 (1 H, m, H-11a);

2.28 and 2.23 (2d, J _AB = 15.4, CH 2 CO 2); 2.16 (m, H- _3'q ); 1.97 (1 H, m, H-6 _A ); 1.9 to 1.1 (5H, m, CH _2); 1.86 (1H, m, _H-6); 1.73 (2H, m, _CH2 -7); 1.14 (3H, s, CH _3); 1.03 (3H, s,

CH ₃ ).

Example 10

Preparation of (-) - cephalotaxyl 2-methoxycarbonylmethyl-6,6-dimethyl-2-tetrahydropyrancarboxylate or anhydrohomoharringtonine from tetrahydropyrancarboxylic acid resulting from Example 6:

1 °) Mixed anhydride method

To a stirred mixture of the hemiester formed in Example 6 (50 mg, 0.22 mmol) in anhydrous toluene (1 mL) at room temperature was added triethylamine (dried over potassium hydroxide) (29.4 µl, 0.22 mmol) and 2. 4,6-trichlorobenzoyl chloride (32.7 µL, 0.22 mmol). The mixture was stirred at 25 ° C for 20 hours (the disappearing starting acid was controlled by IR), 4-dimethylaminopyridine (29 mg, 0.24 mmol) was added, the reaction was allowed to react for 5 minutes, and cephalotaxin (16.5 mL) was added. mg, 0.05 mmol). The mixture was stirred at 25 ° C for 24 hours, then filtered through filter paper and diluted with ether (5 mL). The resulting organic layer was washed successively with water (5 mL), saturated sodium bicarbonate solution (5 mL), again water (5 mL), dried over magnesium sulfate and evaporated in vacuo. The resulting crude product was purified by column chromatography (dichloromethane / methanol (98: 2), silica (15-40 µm)) to give the expected product (16 mg, 56%, two diastereomers). the product obtained in Example 9.

2 °) Method using DCC

To a stirred mixture of the hemiester resulting from Example 6 (100 mg, 0.43 mmol) in anhydrous toluene (1 mL) maintained under an inert atmosphere at room temperature was added 1,3-dicyclohexylcarbodiimide (180 mg, 0.87 mmol). The mixture was stirred for 10 minutes and cephalotaxin (165 mg, 0.52 mmol) and pyrrolidine pyridine (77 mg, 0.52 mmol) were added. Stir the mixture at 18 ± 5 ° C for 18 hours, add ether, filter the reaction on a ground glass filter, and wash the cake with ether. The resulting organic layer was washed successively with 10% sodium bicarbonate solution, water, then dried over magnesium sulphate and evaporated in vacuo. The resulting crude product was purified by column chromatography (dichloromethane / methanol (98: 2), silica (15-40 µm) to give a solid (110 mg, 48%). The product thus obtained showed identical characteristics to the product obtained in Example 9 .

Example 11

4

Preparation of (-) - Cephalotaxyl (2 ' RS) -2-methoxycarbonylmethyl-6,6-dimethyl-2-tetrahydropyrancarboxylate or anhydrohomoharringtonine from ethylene acid resulting from Example 6:

1 °) Method A: via mixed anhydride, coupling by crystallization

To a stirred mixture of the ethylene ester formed in Example 5 (50 mg, 0.22 mmol) in anhydrous toluene (1 mL) at room temperature was added triethylamine (dried over potassium hydroxide) (29 μί, 0.22 mmol) and 2.4 6-trichlorobenzoyl chloride (34 μί, 0.22 mmol). The mixture was stirred at 30 minutes (the disappearing starting acid was controlled by IR), 4-dimethylaminopyridine (30 mg, 0.25 mmol) was added, the reaction was allowed to react for 5 minutes, and cephalotaxin (31 mg, 0.1 mmol) was added. . The mixture was stirred at 18 ± 5 ° C for 16 hours, then filtered through filter paper and diluted with ether (5 mL). The resulting organic layer was washed successively with water (5 mL), saturated sodium bicarbonate solution (5 mL), again water (5 mL), dried over magnesium sulfate and evaporated in vacuo. The resulting crude product is purified by column chromatography (dichloromethane / methanol (98: 2), silica (15 to 40 μιτι) to give the expected product (46 mg, 96%, two 40/60 diastereomers). as the product obtained in Example 9.

2 °) Method using DCC, crystallization coupling

To a stirred mixture of the ethylene acid formed in Example 5 (50 mg, 0.22 mmol) in anhydrous toluene (2 mL) maintained under an inert atmosphere at room temperature was added 1,3-dicyclohexylcarbodiimide (270 mg, 1.31 mmol). . The mixture was stirred for 5 minutes and cephalotaxin (70 mg, 0.22 mmol) and pyrrolidine pyridine (32 mg, 0.22 mmol) were added. The mixture is stirred at 18 ± 5 ° C for 65 hours (the reaction is controlled by CCM, eluent). • «· · · · · · · ·

9 9 9 9 9 9 (9: 9) dichloromethane / methanol 9: 1), the reaction mixture is filtered through a ground glass filter and the cake is washed with toluene (5 ml) and the filtrate is evaporated in vacuo. The resulting crude product was purified by column chromatography (dichloromethane / methanol (98: 2), silica (15-40 µm) to give a solid (40 mg, 35%). The product thus obtained showed identical characteristics to the product obtained in Example 9 .

Example 12

Preparation of purified (-) cephalotaxin from total alkaloid extract of Cephalotaxus sp

°) Preparation of total alkaloid extract

In a 30 liter flask, crushed leaves (fresh or dry) of Cephalotaxus sp (10 mg) are mixed with methanol (20 L) for 65 hours and then sieved (50 L). The solution was filtered and concentrated in vacuo to a volume of 5L. The concentrated solution was acidified with 6% aqueous tartaric acid. Then the hydroalcoholic solution was washed with dichloromethane (5 x 5 L) to remove fatty materials and pigments. The aqueous solution was basified with aqueous ammonia (2.5%) to pH 9, then extracted with dichloromethane (5 x 5 L). After concentration under reduced pressure, the crude alkaloid extract was recovered as white solid crystals (24.5 g). The cephalotaxin content was 71% (HPLC).

2 °) Isolation and chromatographic purification of (-) - cephalotaxin from crude alkaloid extract:

The above crude extract is dissolved in the mobile phase (triethylamine (1.55 / 1000) in deionized water and pH adjusted to 3 with phosphoric acid. The solution is then filtered and injected into a preparative high performance liquid chromatography chromatograph equipped with axial compression and high pressure pump ( stationary phase: n-octadecylsilane, 15 µm, porosity 100, 1 kg) Elution is carried out at a rate of 0.2 l / min The fractions are monitored by UV detector and TLC The remaining fractions are checked by HPLC, then pooled, basified with 2 0.5% aqueous ammonia and extracted with dichloromethane (4 x 400 mL) After concentration under reduced pressure, the obtained resin was triturated with methanol to give (-) - cephalotaxin (18 g) as a white crystalline solid (HPLC purity = 99, The product thus obtained exhibits the following characteristics:

[α] ο ^2θ : -174.1 (c = 0.20; CHCl 3) ¹ H NMR 400 MHz (COCl ₃ ) (δ ppm, J Hz):

6.68 (TH, s, H-17 &lt; + &gt;); 6.65 (TH, s, H-14 &lt; + &gt;); 5.91 and 5.90 (2H, 2d, J _AB = 1.5, OCH ₂ O); 4.93 (1 H, s, H-1); 4.77 (1H, dd, J ₃ -4 = 9.4, J ₃ -oh = 3.4, H-3); 3.73 (3H, s, _OCH3); 3.68 (1H, d, J 4.3 = 9.4, H-4); 3.35 (1H, m, J _AB = 14.3, J = 12.2 and 7.9, H-11β); 3.08 (1 H, m, J =

9.1 and 4.9, H-8a); 2.92 (TH, td, J = 11.6 and 7.1, H-10a); 2.59 (2H, m, H-8a + H-10a); 2.35 (TH, dd, J _AB = 14.4, J = 6.9, H-11a); 2.02 (1H, td, J _AB = 12.1, J = 9.7, H-6 _A ); 1.87 (1H, m, J _AB = 12.1, J = 7.9 and 4.4, H- _6B ); 1.74 (2H, m, _CH2 -7); 1.62 (1H, d, J ₃ , _OH = 3.5, 3-OH).

Example 13

Preparation of lithium alcoholate (-) cephalotaxin (captured as 3-O-acetyl derivative)

1 °) Butyllithium method

A commercial solution of butyllithium in hexane (0.44 mL, 1.6 M in hexane, 0.70 mmol) was added to a stirred mixture of (-) - cephalotaxin (200 mg, 0.63 mmol) in anhydrous tetrahydrofuran (6.8 mL). ). The reaction mixture is maintained at -60 ° C for 20 minutes, then at -48 ° C for 30 minutes, acetic anhydrides (90 µL, 0.095 mmol) are added over 8 minutes and the mixture is maintained at -48 ° C. for 20 minutes and then at 0 ° C for 1 hour. The mixture was quenched with ammonium chloride solution (5 mL) and then extracted with ethyl acetate (3 x 8 mL). The combined organic layers were washed with brine (15 mL), dried over magnesium sulfate and evaporated to dryness. Resulting raw ···· · 99 ··

99999

999 9 · 9 9 9 9 9

The product was purified by column chromatography (dichloromethane / methanol (98: 2) silica (15-40 µm) 6 g) to give a white solid (60 mg, 26%). the product thus obtained exhibits the following characteristics:

¹ H NMR 400 MHz (CDCl ₃ ) (δ ppm, J Hz):

6.60 (1 H, s, H-17); 6.57 (1 H, s, H-14); 5.89 and 5.86 (2H, 2d, J _AB = 1.4, OCH ₂ O); 5.80 (1H, d, J _4.3 = 9.3, H-3); 5.05 (1 H, s, H-1); 3.77 (1H, d, = 9.4, H-4); 3.72 (3H, s, _OCH3); 3.23 (1H, m, J _A b = 14.3, J = 12.3 and 7.9, β-11β); 3.08 (1 H, m, H-8a); 2.92 (1H, td, J = 11.5 and 7.1, H-10a); 2.57 (2H, m, Η-8β + Η-10β) _; 2.36 (1H, dd, J _AB = 14.4, J = 7.0, H-11a); 2.02 (1H, td, J _AB = 12.1, J = 9.7, H-6 _A ); 1.88 (1H, m, J _AB = 12.1, J = 8.0 and 4.0, _H-6); 1.74 (2H, m, _CH2 -7), 1.57 (3H, s, OAc).

2 °) Lithium bis- (trimethylsilyl) amide (LHDS) method

A commercial solution of 1M lithium bis- (trimethylsilylamide) in tetrahydrofuran (0.95 mL, 0.95 mmol) was added at -40 ° C to a stirred solution of (-) - cephalotaxin (200 mg, 0.63 mmol) in anhydrous tetrahydrofuran. The mixture was stirred for 5 minutes, acetic anhydride (90 µL, 0.95 mmol) was added and the reaction was worked up in a similar manner as above at 1 °). The product thus obtained exhibits identical characteristics to the product obtained in the n-butyllithium method.

3 °) Lithium diisopropylamide (LDA) method

A commercial solution of 2M lithium diisopropylamide in tetrahydrofuran (0.35 mL, 0.70 mmol) was added over 20 minutes at -60 ° C to a stirred solution of (-) - cephalotaxin (200 mg, 0.63 mmol) in anhydrous tetrahydrofuran (6, 8 ml). The mixture was stirred at -60 ° C for 20 minutes, then at -48 ° C for 30 minutes and acetic anhydride (90 µL, 0.95 mmol) was added. The solution was stirred at -48 ° C for 20 minutes and then at 0 ° C for 1 hour and the reaction mixture was worked up according to Method 1) above. The product obtained has the same characteristics as the product prepared by the butyllithium method.

4 °) Sodium hydride method ··

To a stirred mixture of sodium hydride (1.5 g) in freshly distilled dimethylformamide (3 mL) at -60 ° C was added a solution of cephalotaxin (200 mg, 0.63 mmol) in dimethylformamide (3 mL) and acetic anhydride (90 μΙ, 0 , 95 mmol). After stirring at ambient temperature for 24 hours, the reaction mixture was treated at 0 ° C with water (3 mL) and extracted with ether (3 x 5 mL). The combined organic layers were dried over magnesium sulfate and evaporated in vacuo. The product obtained exhibits identical characteristics to the product obtained in the butyl lithium method.

Example 14

Preparation of (-) - cephalotaxyl 2-methoxycarbonylmethyl-6,6-dimethyl-2-tetrahydropyran carboxylate or anhydrohomoharringtonine via cephalotaxin lithium alcolate:

To a stirred solution of (-) - cephalotaxin lithium alcolate (158 mg, 0.5 mmol) in anhydrous tetrahydrofuran prepared according to Example 13 was added at -50 ° C over 10 minutes the mixed anhydride resulting from Example 10 (0.75 mmol). Stir the mixture at -50 ° C for 30 minutes, then at 0 ° C for 2 hours, quench the reaction mixture with saturated ammonium chloride solution (5 mL) and extract with ethyl acetate (3 x 10 mL). The combined organic layers were washed with brine (15 mL), dried over magnesium sulfate and evaporated in vacuo. The resulting crude product was purified by column chromatography (dichloromethane / methanol (98: 2), silica (15-40 μΓη) 7 g) to give a white solid (48 mg). The product obtained has the same characteristics as the product obtained in Example 9.

Example 15

Preparation of the diastereomeric mixture of (-) - quinidyl 2-methoxycarbonylmethyl-6,6-dimethyl-2-tetrahydropyrancarboxylates from tetrahydropyrancarboxylic acid resulting from Example 6:

1 °) Method A: via mixed anhydride

To a stirred mixture of the acid formed in Example 6 (458 mg, 1.99 mmol) in anhydrous toluene (8 mL) at room temperature was added triethylamine (dried over potassium hydroxide) (270 µL, 1.92 mmol) and 2.4, 6-trichlorobenzoyl chloride (300 µL, 1.91 mmol). The mixture was stirred for 3 hours (the disappearing starting acid was controlled by IR), 4-dimethylaminopyridine (352 mg, 2.88 mmol) was added, the reaction was allowed to react for 5 minutes, and quinine (936 mg, 2.88 mmol) was added. The mixture was stirred at 18 ± 5 ° C for 65 hours, then filtered through filter paper and diluted with ether (15 mL). The resulting organic layer was washed successively with water (15 mL), saturated sodium bicarbonate solution (15 mL), again water (15 mL), dried over magnesium sulfate and evaporated in vacuo. The resulting crude product was purified by column chromatography (dichloromethane / methanol (99: 1), silica (15-40 µm) to give the expected product (930 mg, 84%, two 50/50 diastereomers). :

¹ H NMR 400 MHz (CDCl ₃ ) (δ ppm, J Hz):

8.73 (1H, m, H- _qn ), 8.0 and 7.98 (1H, 2d, J = 9.2, H-8 _qn ); 7.63 and 7.50 (1H, 2br.s); 7.45 (br. S) and 7.39 (d, J = 4.5) (1H, H-3 _qn ); 7.36 (1H, dd, J = 9.1 and 2.6, H-7 _qn ); 6.50 (1 H, br.s); 5.89 (1H, m, = CH _qn ); 5.03 (2H, m, = _CH2qn ); 3.99 and 3.97 (3H, 2s, OCH ₃ ); 3,54 a

3.33 (3H, 2br.s, _OCH3); 3.2 to 1.0 (m, 7 x _CH2 + 3CH); 2,92 and 2,67 (2d, J _AB = 14,9, φ φ φ φ φ · · φ φ

φφφ

CH2CO2); 2.87 (d, Jab = 14.8, CH 2 CO 2); 1.17 and 0.99 (2s, 2 x CH ₃ ); 1.03 and 0.42 (2br.s., 2xCH _3).

2 °) Method B: DCC

To a stirred mixture of the tetrahydrocarboxylic acid formed in Example 6 (200 mg, 0.87 mmol) in anhydrous toluene (4 mL) maintained under an inert atmosphere at room temperature was added 1,3-dicyclohexylcarbodiimide (239 mg, 1.16 mmol). The mixture was stirred for 5 minutes, quinine (94 mg, 0.29 mmol) and pyrrolidine pyridine (43 mg, 0.29 mmol) were added. The mixture was stirred at 18 ± 5 ° C for 65 hours (reaction monitored by CCM, eluent dichloromethane / methanol; 9: 1), the reaction mixture was filtered on a ground glass filter, and the cake was washed with toluene (5 mL) and the filtrate was washed. Evaporate in vacuo. The resulting crude product was purified by column chromatography (dichloromethane / methanol (9: 1), silica (15-40 µm)) to give the expected product (96 mg, 60%, two 50/50 diastereomers). The product thus obtained exhibits identical characteristics to the product obtained above.

Example 16

Preparation of (-) - menthyl 2-methoxycarbonylmethyl-6,6-dimethyl-2-tetrahydropyrancarboxylate, tetrahydropyrancarboxylic acid resulting from Example 6:

To a stirred mixture of the acid formed in Example 6 (50 mg, 0.22 mmol) in anhydrous toluene (1 mL) maintained under an inert atmosphere at room temperature was added 1,3-dicyclohexylcarbodiimide (90 mg, 0.44 mmol). The mixture was stirred for 5 minutes, (-) 88 menthol (68 mg, 0.44 mmol) and pyrrolidinopyridine (64 mg, 0.44 mmol) were added. The mixture was stirred at 30 ° C for 1 hour and then at 8 ° C for 15 hours (reaction controlled by CCM, eluent dichloromethane / methanol; 9: 1), the reaction mixture was filtered on a ground glass filter and the cake was washed with toluene (5 mL). and the filtrate was evaporated in vacuo. The resulting crude product was purified by column chromatography (cyclohexane / ethyl acetate (95: 5 then 90:10), silica (15-40 µm) 4 g) to give the expected product (40 mg, 50%, two 60/40 diastereomers). . The product thus obtained exhibits the following characteristics:

¹ H NMR 400 MHz (CDCl ₃ ) (δ ppm, J Hz):

4.68 (1H, m, H-1 _mo m); 3.64 (3H, s, _OCH3); 2.84 and 2.64 (2d, J _AB = 14.6, CH 2 CO 2); 2.83 and 2.63 (2d, J _AB = 14.3, _{CH? CO);} 2.29 (1H, m, H- _30q ); 2.1 to 0.8 (m, CH and _CH2); 1.21 (3H, 2s, CH _3); 1.17 and 1.16 (3H, 2s, CH ₃ ); 0.9 and 0.88 (6H, 2d, J = 6.4, 2 * _CH3mon ); 0.74 and 0.72 (3H, 2d, J = 6.8, _CH3mon ).

Example 17

Preparation of (-) - 2-methoxycarbonylmethyl-6,6-dimethyl-2-tetrahydropyrancarboxylate (-) methylmandelate from tetrahydropyrancarboxylic acid resulting from Example 6:

To a stirred mixture of tetrahydropyranecarboxylic acid formed in Example 6 (226 mg, 0.98 mmol) in anhydrous toluene (4 mL) maintained under an inert atmosphere at room temperature was added 1,3-dicyclohexylcarbodiimide (261 mg, 1.2 mmol). The mixture was stirred for 5 minutes and menthylmandelate (53 mg, 0.32 mmol) and pyrrolidinopyridine (47 mg, 0.32 mmol) were added. The mixture was stirred at 18 ± 5 ° C for 12 hours, then the reaction mixture was filtered through a ground filter, the cake was washed with toluene (5 mL), and the filtrate was evaporated in vacuo. The resulting crude product was purified by column chromatography (dichloromethane / methanol (9: 1), silica (15 to 40 µm)) to give a colorless oil (64 mg, 17%, two diastereomers).

¹ H NMR 400 MHz (CDCl ₃ ) (δ ppm, J Hz):

7.47 (2 H, m, Ph); 7.38 (3 H, m, Ph); 5.96 (1 H, s, CH); 3.73 and 3.72 (3H, 2s, OCH ₃ ); 3.54 (3H, 2s, _OCH3); 2.88 and 2.72 (2d, J _A b = 14.4 (CH 2 CO 2); 2.85 and 2.65 (2d, J _AB =

14.2, CH 2 CO 2); 2.35 (1H, m, H- _30q ); 2.0- 1.15 (5H, m, CH _2); 1.23 and 1.22 (3H, 2s, CH ₃ ); 1.19 and 1.07 (3H, 2s, CH ₃ ).

Example 18

Separation of (-) - quinyl (2'R) -anhydrohomoharringtonate and (-) - quinyl (2'S) anhydrohomoharringtonate from the diastereomeric mixture resulting from Example 15:

The diastereomeric mixture of (-) - quinyl (2'R) -anhydrohomoharringtonate and (-) - quinyl (2'S) anhydrohomoharringtonate (5 g) was subjected to preparative HPLC. The above mixture is dissolved in a buffer (triethylamine (1.55 / 1000)) in deionized water and orthophosphoric acid to adjust the pH to 3. The solution is then filtered and injected into a preparative high performance liquid chromatography chromatograph equipped with axial compression and high pressure pump (stationary). phase: noctadecylsilane, 15 μηη, porosity 100, 1 kg; mobile phase: buffer / acetonitrile 70/30). Elution was performed at a rate of 0.2 L / min. The fractions were monitored by U.V. detector and TLC. The remaining fractions were checked by HPLC, then combined, basified with 2.5% aqueous ammonia and extracted with dichloromethane (4 x 400 mL). After concentration under reduced pressure, two separated isomers were obtained as a white crystalline solid corresponding to (-) - quinyl (2'R) -anhydrohomoharringtonate (2 g) and (-) - quinyl (2'S) anhydrohomoharringtonate (2.2 g) . The products thus obtained exhibit the following characteristics:

1 °) Diastereomer 2 ·

IR (NaCl film) (cm ^-1 ): 2947; 2871; 1743; 1626; 1509.

¹ H NMR 400 MHz (CDCl ₃ ) (δ ppm, J Hz):

8.73 (1H, d, J = 4.4, H-2 _qn ), 8.0 (1H, d, J = 9.2, H-8 _qn ); 7.50 (1 H, br.s); 7.39 (1H, d, J = 4.5, H-3 _qn ); 7.36 (1H, dd, H-7 _qn ); 6.39 (1 H, br.s); 5.88 (1H, m, = CH _qn ); 5.03 (2H, m, = _CH2qn ); 3.97 (3H, s, _OCH3); 3.31 ((3H, br.s, OCH ₃ ); 3.5 to 1.2 (m, 7 x CH ₂ + CH 3);

2.86 and 2.64 (2H, 2d, J _A b = 15.0, CH 2 CO 2); 1.17 (3H, s, CH _3); 0.99 (3H, s, _CH3).

2 °) Diastereomer 2'S

IR (NaCl film) (cm ^-1 ): 2947; 2871; 1743; 1626; 1509.

¹ H NMR 400 MHz (CDCl 3) (δ ppm, J Hz):

8.74 (1H, d, J = 4.4, H-2 _qn ); 7.99 (1H, d, J = 9.2, H-8 _qn ); 7.65 (1H, br.s, H-3 _qn ); 7.44 (1H, br. S, H-5 _qn ); 7.36 (1H, dd, J = 9.2 and 2.7, H-7 _qn ); 6.55 (1 H, br.s); 5.89 (1H, m, = CH _qn ); 5.05 (2H, m, = _CH2qn ); 3.99 (3H, s, _OCH3); 3.54 (3H, s, _OCH3); 3.1 - 1.0 (m, 7 x CH ₂ + 3 CH); 2.91 and 2.67 (2H, 2d, J _AB = 15.0, CH 2 CO 2); 1.03 <3H, br s, CH _3); 0.44 (3H, br. _S , CH ₃ ).

Example 19

Preparation of (2R) -anhydrohommoharringtonic acid from (-) - quinyl (2'R) anhydrohomoharringtonate resulting from Example 18:

1 °) Obtained by hydrogenolysis

To a stirred solution of chinyl (2'R) -anhydroharringtonate (100 mg, 0.19 mmol) in ethyl acetate (11 mL) was added 10% palladium on charcoal (40 mg). The resulting mixture was stirred at room temperature under hydrogen pressure (50 psi) for 20 hours, and after CCM control, the reaction mixture was filtered and the resulting organic layer was treated with saturated sodium bicarbonate solution. The aqueous layer was washed with ethyl acetate and, after acidification with 1 N hydrochloric acid, extracted with ethyl acetate. The combined organic layers were washed with brine, dried over magnesium sulfate and evaporated to dryness to give a yellow solid (20 mg, 50%). The product thus obtained exhibits the following characteristics:

[α] _D ²⁰ : -23 (c = 0.38; CHCl ₃ ).

IR (NaCl film) (cm ^-1 ): 2974; 2951; 1740; 1718; 1437.

¹ H NMR 400 MHz (CDCl ₃ ) (δ ppm, J Hz):

^{The 1} H NMR spectra of the product thus obtained are identical to those described in Example 6-1.

2) Obtained by total saponification and then selective methylation

A mixture of potassium hydroxide (396 mg, 7.1 mmol) in water (8 mL) was added to a stirred solution of chinyl (2'R) -anhydroharringtonate (396 g, 0.72 mmol) in ethanol (15 mL). The mixture was stirred at reflux for 24 hours. After cooling to room temperature and removing the ethanol in vacuo, the residue was treated with water (10 mL) and the resulting aqueous layer was extracted with ether (4 x 15 mL). After acidification (pH 1) with 2N hydrochloric acid and saturation with sodium chloride, the aqueous layer was extracted with ethyl acetate (3 x 15 mL). The combined organic layers were dried over magnesium sulfate and evaporated to dryness to give a yellow solid (110 mg, 72%). The intermediate dibasic acid thus obtained exhibits the following characteristics:

[α] D ²⁰ : -14 (c = 0.54; CHCl ₃ ).

IR (NaCl film) (cm ^-1 ): 2975; 2941; 1716; 1217.

¹ H NMR 400 MHz (CDCl ₃ ) (δ ppm, J Hz):

^{The 1} H NMR spectra of the product thus obtained are identical to those described in Example 4.

A mixture of the (2R) -cyclic dibasic acid mentioned above (110 mg, 0.5 mmol) and a technical solution of boron trifluoride-methanol complex in methanol (1.1 mL, BF ₃ 12% w / w) is stirred at 18 ± 5 ° C for 15 hours. After carefully adding the reaction mixture to saturated sodium bicarbonate solution (20 mL), the resulting aqueous layer was washed with ether (3 x 15 mL), acidified (pH 1) with 2N hydrochloric acid, and extracted with ether (3 x 15 mL). The combined organic layers were dried over magnesium sulfate and evaporated to dryness to give a viscous yellow oil (69 mg, 59%). The product thus obtained exhibits the same characteristics as the product according to method 1 above.

Example 20

Preparation of (2S) -anhydrohommoharringtonic acid from (-) - quinyl (2'R) anhydrohomoharringtonate resulting from Example 18:

9 4 4 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25

4 · ««

1 °) Obtained by hydrogenolysis

To a stirred solution of chinyl (2'S) -anhydroharringtonate (100 mg, 0.19 mmol) in ethyl acetate (11 mL) was added 10% palladium on charcoal (40 mg). The resulting mixture was stirred at room temperature under hydrogen pressure (50 psi) for 20 hours. After CCM control, the reaction mixture was filtered and the resulting organic layer was treated with saturated sodium bicarbonate solution. The aqueous layer was washed with ethyl acetate and, after acidification with 1 N hydrochloric acid, extracted with ethyl acetate. The combined organic layers were washed with brine, dried over magnesium sulfate and evaporated to dryness to give a yellow solid (23 mg, 53%). The product thus obtained exhibits the following characteristics:

[α] ₀ ^2θ : +30 (c = 0.36; CHCl ₃ ).

IR (NaCl film) (cm ^-1 ): 2975; 2951; 1740; 1718; 1439.

¹ H NMR 400 MHz (CDCl 3) (δ ppm, J Hz):

^{The 1} H NMR spectra of the product thus obtained are identical to those described in Example 6-1.

2) Obtained by total saponification and then by selective methylation

A mixture of potassium hydroxide (430 mg, 7.7 mmol) in water (9 mL) was added to a stirred solution of chinyl (2'S) -anhydroharringtonate (447 g, 0.81 mmol) in ethanol (16 mL) and the resulting mixture was stirred at 24 hours. After cooling to room temperature and removing the ethanol in vacuo, the residue was treated with water (10 mL) and the resulting aqueous layer was extracted with ether (4 x 15 mL). After acidification (pH 1) with 2N hydrochloric acid and saturation with sodium chloride, the aqueous layer was extracted with ethyl acetate (3 x 15 mL). The combined organic layers were dried over magnesium sulfate and evaporated to dryness to give a yellow solid (140 mg, 80%). The intermediate dibasic acid thus obtained exhibits the following characteristics:

• • · · · · <· [a] _D ^20: +8 (c = 0.19, CHCl _3).

IR (NaCl film) (cm ^-1 ): 2975; 2945; 1717.

¹ H NMR 400 MHz (CDCl ₃ ) (δ ppm, J Hz):

^{The 1} H NMR spectra of the product thus obtained are identical to those described in Example 4.

The mixture of the (2S) -cyclic dibasic acid mentioned above (136 mg, 0.62 mmol) and a technical solution of boron trifluoride-methanol complex in methanol (1.3 mL, BF ₃ 12% w / w) is stirred at 18 ± 5 ° C for 15 hours. After carefully adding the reaction mixture to saturated sodium bicarbonate solution (20 mL), the resulting aqueous layer was washed with ether (3 x 15 mL), acidified (pH 1) with 2N hydrochloric acid, and extracted with ether (3 x 15 mL). The combined organic layers were dried over magnesium sulfate and evaporated to dryness to give a viscous yellow oil (81 mg, 63%). The product thus obtained exhibits the same characteristics as the product according to method 1 above.

Example 21

Preparation of anhydrohomoharringtonine by esterification of cephalotaxin with (2R) - (+) anhydrohomoharringtoninic acid

To a stirred mixture of the hemiester (R) formed in Example 19 (65 mg, 0.28 mmol) in anhydrous toluene (1 mL) at room temperature was added triethylamine (dried over potassium hydroxide) (38 μΙ, 0.28 mmol) and 2 mL. , 4,6-trichlorobenzoyl chloride (43 μΙ, 0.28 mmol). The mixture was stirred at 30 ° C for 1.5 hours (control of vanishing starting acid IR), dimethylaminopyridine (50 mg, 0.41 mmol) was added and the reaction was allowed to react for 5 minutes and cephalotaxin (129 mg, 0) was added. , 41 mmol). The mixture was stirred at 30 ° C for 18 hours, then the reaction mixture was filtered through filter paper and diluted with ether (5 mL). The resulting organic layer was washed successively with water (5 mL), saturated sodium bicarbonate solution (5 mL), again with water, then dried over magnesium sulfate and evaporated in vacuo. The resulting crude product was purified by column chromatography (dichloromethane / methanol (99: 1), silica (15-40 µm)) to give a solid (65 mg, 43%). The product thus obtained exhibits the following characteristics:

¹ H NMR 400 MHz (CDCl ₃ ) (δ ppm, J Hz):

6.61 (1 H, s, H-17 *); 6.58 (1 H, s, H-14); 5.92 (1H, d, J ₃ _ ₄ = 9.6, H-3); 5.87 and 5.79 (2H, 2s, OCH ₂ O); 5.04 (1H, br. S, H-1); 3.80 (1H, d, = 9.2, H-4); 3.70 (3H, s, _OCH3); 3.59 (3H, s, _OCH3); 3.12 (2H, m, H-11β + H-8a); 2.95 (1 H, m, H-10a); 2.60 (2H, m, Η-8β + Η-β); 2.38 (1H, m, H-1 [alpha]); 2.13 and 1.66 (2H, 2d, J AB ⁼ 14.3, _CH? CO); 2.02 (1 H, m, H-6 _A ); 1.90 (1 H, m, H- _6B ); 1.76 (2H, m, CH ₂ -7), 1.8-1.2 (6H, 'm, 3 x _CH2);

1.10 (3H, s, CH ₃ ).

Example 22

Preparation of anhydrepihomoharringtonine by esterification of cephalotaxin with (2S) - (+) anhydrohomoharringtoninic acid

· Ί · · <<<<<<<

9 9 '9 9 9 9 «·» · · · · · · · · ·

Κ triethylamine (dried over potassium hydroxide) (52 µL, 0.38 mmol) was added to a stirred mixture of the hemiester (S) formed in Example 20 (87 mg, 0.38 mmol) in anhydrous toluene (1.7 mL) at room temperature. and 2,4,6-trichlorobenzoyl chloride (57 µL, 0.38 mmol). The mixture was stirred at 30 ° C for 1.5 hours (control of vanishing starting acid IR), dimethylaminopyridine (70 mg, 0.57 mmol) was added and the reaction was allowed to react for 5 minutes and cephalotaxin (180 mg, 0) was added. , 57 mmol). The mixture was stirred at 30 ° C for 18 hours, then the reaction mixture was filtered through filter paper and diluted with ether (5 mL). The resulting organic layer was washed successively with water (5 mL), saturated sodium bicarbonate solution (5 mL), again with water, then dried over magnesium sulfate and evaporated in vacuo. The resulting crude product was purified by column chromatography (dichloromethane / methanol (99: 1), silica (15-40 µm)) to give a solid (101 mg, 50%). The product thus obtained exhibits the following characteristics:

¹ H NMR 400 MHz (CDCl 3) (δ ppm, J Hz):

6.61 (1 H, s, H-17 *); 6.57 (1 H, s, H-14 *); 5.84 (3H, m, H-3) + OCH ₂ O); 5.04 (1 H, H-1); 3.78 (1H, d, J 4-3 = 9.7, H-4); 3.65 (3H, s, _OCH3); 3.59 (3H, s, _OCH3); 3.23 (1 H, m, H 11β); 3.09 (1 H, m, H-8a); 2.93 (1 H, m, H-10a); 2.58 (2H, m, Η-8β + Η-10β); 2.39 (1H, dd, Jab = 14.4, J = 7.0, H-11a); 2.16 and 1.83 (2H, 2d, J _AB = 14.5, CH ₂ CO ₂ ); 2.06 (1H, m, _H-6); 1.88 (1 H, m, H-6 _B ); 1.74 (2H, m, CH ₂ -7), 1.5-1.2 (6H, m, 3 x _CH2);

1.11 (3H, s, CH _3); 1.02 (3H, s, CH ₃ ).

Example 23

Preparation of 6-bromo-6'-desoxy-homoharringtonine from anhydrohomoharringtonine produced in Example 21:

>> ·> · »» »» »· · · · · · · · · · · · · · · · · · ·» »»

To a stirred solution of the product formed in Example 21 (60 mg, 0.114 mmol) in anhydrous dichloromethane (300 μΐ) was added a technical solution of hydrobromic acid in acetic acid (205 µL, 1.02 mmol, HBr 30 wt. % (w / w).

The mixture was stirred at -10 ° C for 3 hours and saturated sodium bicarbonate solution was added to pH 8. The resulting aqueous layer was extracted three times with dichloromethane and the combined organic layers were dried over magnesium sulfate and evaporated to dryness to give a yellow oil (60 mg). , 87%). The crude product obtained has the following characteristics:

IR (ATR) (cm ^-1 ): 2957; 1744; 1653; 1487; 1223.

¹ H NMR 400 MHz (CDCl 3) (δ ppm, J Hz):

6.63 (1 H, s, H-17); 6.54 (1 H, s, H-14 *); 5.99 (1H, d, J ₃ _ ₄ = 9.8, H-3); 5.87 (2H, m, OCH ₂ O); 5.05 (1H, s H-1); 3.78 (1H, d, J 4-3 = 9.8, H-4); 3.69 (3H, s, _OCH3); 3.58 (3H, s, OCH 3); 3.54 (1H, s, 2 &apos;-OH); 3.10 (2H, m, J-11β + H-8a); 2.94 (1 H, m, H-10a);

2.60 (2H, m, Η-8β + Η-10β); 2.39 (1H, dd, J _AB = 14.0, J = 6.8, H-11a); 2.26 and 1.89 (2H, 2d, J _AB = 16.5, PDO2); 2.03 (1 H, m, H-6 _A ); 1.91 (1 H, m, H- _6B ); 1.75 (2H, m, CH ₂ -7), 1.74 (3H, s, CH ₃ ); 1.72 (3H, s, CH _3); 1.6 to 1.2 (6H, m, 3 x _CH2).

Example 24

Preparation of 6'-bromo-6'-desoxy-epihomoharringtonine from anhydroepihomoharringtonine produced in Example 22:

To a stirred solution of the product formed in Example 22 (60 mg, 0.114 mmol) in anhydrous dichloromethane (300 μΙ) was added a technical solution of hydrobromic acid in acetic acid (205 μΙ, 1.02 mmol, HBr 30 wt. % (w / w). The mixture was stirred at -10 ° C for 3 hours and saturated sodium bicarbonate solution was added to pH 8. The resulting aqueous layer was extracted three times with dichloromethane and the combined organic layers were dried over magnesium sulfate and evaporated to dryness to give a yellow oil (63 mg). , 91%). The crude product obtained has the following characteristics:

IR (ATR) (cm ^-1 ): 2957; 1744; 1653; 1487; 1223.

¹ H NMR 400 MHz (CDCl 3) (δ ppm, J Hz):

6.64 (1 H, s, H-17); 6.59 (1 H, s, H-14); 5.97 and 5.87 (2H, d, J _AB = 1.1, OCH ₂ O); 5.95 (1H, d, J 3-4 = 9.7, H-3); 5.04 (1H, s H-1); 3.78 (1H, d, J 4-3 = 9.7, H-4); 3.67 (3H, s, OCH 3); 3.66 (3H, s, OCH 3); 3.49 (1 H, s, 2-OH); 3.10 (2H, m, J-11β + H-8a); 2.93 (1 H, m, H-10a); 2.62 and 2.54 (2H, 2d, J _AB = 16.5, CH _{2 CO);} 2.60 (2H, m, Η-8β + H10β); 2.40 (1H, m, H-11a); 2.03 (1 H, m, H-6 _A ); 1.89 (1 H, m, H- _6B ); 1.74 (2H, m, _CH2 -7); 1.72 (3H, s, CH _3); 1.72 (3H, s, CH _3); 1.70 (3H, s, CH ₃ ); 1.6 to 0.7 (6H, m, 3 x _CH2).

Example 25

Preparation of homoharringtonine from 6'-bromo-6'-desoxyhomoharringtonine produced in Example 23

1. Method A:

• J # 1 * ·· φ φ φ φ · · · · · · φ φ <

A 5% solution of sodium bicarbonate (3 mL) was added to a stirred solution of the product of Example 23 (60 mg, 0.099 mmol) in acetone (1.5 mL). The mixture was stirred at room temperature for 2 hours, then evaporated to dryness in vacuo and the remaining aqueous layer was extracted three times with dichloromethane. The combined organic layers were dried over magnesium sulfate and evaporated to dryness. The resulting crude product (55 mg) was purified by column chromatography (dichloromethane, then dichloromethane / methanol (99: 1 then 95: 5), silica (15 to 40 µm) 2.75 g) to give homoharringtonine (29 mg, 47 %). The product thus obtained exhibits the following characteristics:

[α] _D ²⁰ : -110 (c = 0.24; CHCl ₃ ).

IR (NaCl film) (cm ^-1 ): 3468; 2961; 1656; 1487; 1224; 1033.

¹ H NMR 400 MHz (CDCl ₃ ) (δ ppm, J Hz):

6.63 (1 H, s, H-17); 6.55 (1H, s, H-14 '); 6.01 (1H, d, J ₃ = 9.8, H-3); 5.87 (2H, m, OCH ₂ O); 5.05 (1H, s H-1); 3.78 (1H, d, J 4-3 = 9.6, H-4); 3.68 (3H, s, _OCH3); 3.58 (3H, s, _OCH3); 3.54 (1H, s, 2 &apos;-OH); 3.10 (2H, m, H-11β + H-8a); 2.95 (1 H, m, H-10a); 2.59 (2H, m, Η-8β + Η-10β); 2.38 (1H, d, J _AB = 14.0, J = 6.7, H-11a); 2.27 and 1.90 (2H, 2d, J _AB = 16.5, CH7CO7); 2.02 (1 H, m, H-6 _A ); 1.90 (1 H, m, H- _6B ); 1.76 (2H, m, _CH2 -7); 1.5- 1.15 (6H, m, 3 x _CH2); 1.30 (1 H, s, 6-OH); 1.19 (6H, 2s, 2 x _CH3).

2) Method B:

Saturated calcium carbonate solution (3 mL) was added to a stirred solution of the product formed in Example 23 (60 mg, 0.099 mmol) in acetone (3 mL). The mixture was stirred at room temperature for 2 hours to give the same product as in Method A (CCM specified).

2) Method C:

A saturated solution of calcium carbonate (9 mL) was added to a stirred solution of the product formed in Example 23 (60 mg, 0.099 mmol) in acetone (3 mL). The mixture was stirred at room temperature for 2 hours to give the same product as in Method A (CCM specified).

? · **

100 ALIGN!

4) Method D:

To a stirred solution of the product of Example 23 (60 mg, 0.099 mmol) in acetone / water (3/2, 2.15 mL) was added silver nitrate (25 mg, 0.149 mmol). The mixture was stirred at room temperature for 2 hours to give the same product as in the method

A (CCM specified).

Example 26

Preparation of epihomoharringtonine:

1 °) from the 6'-bromo-6'-desoxy-epihomoharringtonine formed in Example 24:

(a) Method A:

A 5% solution of sodium bicarbonate (3 mL) was added to the stirred solution of Example 24 (60 mg, 0.099 mmol) in acetone (1.75 mL). The mixture was stirred at room temperature for 2 hours, the reaction mixture was evaporated in vacuo and the remaining aqueous layer was extracted three times with dichloromethane. The combined organic layers were dried over magnesium sulfate and evaporated to dryness. The resulting crude product (60 mg) was purified by column chromatography (dichloromethane, then dichloromethane / methanol (99: 1 then 97: 3), silica (15-40 µm) 3 g) to give epihomoharringtonine (29 mg, 47%). . The product thus obtained has the following characteristics:

[α] _D ²⁰ : -92 (c = 0.29; CHCl ₃ ).

IR (NaCl film) (cm ^-1 ): 3514; 2961; 1744; 1655; 1488; 1223; 1035.

101 ¹ H NMR 400 MHz (CDCl ₃ ) (δ ppm, J Hz):

6.65 (1 H, s, H-17); 6.60 (1 H, s, H-14 *); 5.95 (1H, d, H-3), 5.95 and 5.86 (2H, 2d,

OCH 2 O); 5.05 (1H, s H-1); 3.78 (1H, d, J 4-3 = 9.7, H-4); 3.68 (3H, s, _OCH3); 3.66 (3H, s, OCH 3); 3.52 (1H, br. S, 2-OH); 3.13 (2H, m, H-11β + H-8a); 2.97 (1 H, m, H-10a); 2.63 (2H, m, Η-8β + Η-10β); 2.61 and 2.52 (2H, 2d, J _AB = 16.5, CH 2 CO 2); 2.40 (1H, dd, J _AB = 13.8, J = 6.3, H-11a); 2.04 (1 H, m, H-6 _A ); 1.94 (1 H, m, H- _6B ); 1.78 (2H, m, _CH2 -7); 1.45 to 0.7 (6H, m, 3 x _CH2); 1.16 (3H, s, CH _3); 1.15 (3H, s, _CH3).

2) Method B:

A saturated solution of calcium carbonate (3 mL) was added to a stirred solution of the product formed in Example 24 (60 mg, 0.099 mmol) in acetone (3 mL). The mixture was stirred at room temperature for 2 hours to give the same product as in Method A (CCM specified).

2) Method C:

Saturated solution of calcium carbonate (9 mL) was added to a stirred solution of the product of Example 24 (60 mg, 0.099 mmol) in acetone (3 mL). The mixture was stirred at room temperature for 2 hours to give the same product as in Method A (CCM specified).

4) Method D:

To a stirred solution of the product of Example 24 (60 mg, 0.099 mmol) in acetone / water (3/2, 2.15 mL) was added silver nitrate (25 mg, 0.149 mmol). The mixture was stirred at room temperature for 2 hours to give the same product as in Method A (CCM specified).

2 °) from the anhydroepihomoharringtonine formed in Example 22

To a stirred solution of the anhydroepihomoharringtonine formed in Example 22 (58 mg, 0.109 mmol) in anhydrous dichloromethane (0.3 mL) was added a technical solution of hydrobromic acid in acetic acid (0.195 mL, 0.98 mmol, HBr 30%) at -10 ° C. w / w). The mixture was stirred at 0 ° C for 3 hours, water (2.8 mL) was added and then the temperature was raised to 20 ° C. The mixture was stirred at 20 ° C for 3 hours, added

102 sodium bicarbonate solution (0.76M, 6 mL) to pH 8. The resulting aqueous layer was saturated with sodium chloride, extracted with dichloromethane (3 x 10 mL), and the combined organic layers were dried over magnesium sulfate and evaporated to dryness to give epihomoharringtonine (45 mL). mg, dry, 75%). The crude product thus obtained shows the same characteristics as the product obtained by Method A.

Example 27

Preparation of homoharringtonine for pharmaceutical use from crude semi-synthetic homoharringtonine resulting from Example 25 by preparative high performance liquid chromatography:

homoharringtonine

°) Method A

Crude homoharringtonine (35 g) is dissolved in a buffer (triethylamine (1.55 / 1000) in deionized water and the pH is adjusted to 3 with phosphoric acid. The solution is then filtered and injected into a preparative high performance liquid chromatography chromatograph equipped with axial compression and high pressure pump. (stationary phase: n-octadecylsilane, 15 µm, porosity 100, 1 kg; mobile phase: buffer / tetrahydrofuran 85/15) .Elution is performed at a rate of 0.2 l / min. The fractions are monitored by UV detector and TLC. The reaction mixture was concentrated by vacuum, then combined, basified with 2.5% aqueous ammonia, and extracted with dichloromethane (4 x 400 mL). Concentration under reduced pressure gave homoharringtonine as a pale yellow resin which was triturated in 8/2 water / water. methanol and pure

103 homoharringtonine as a white crystalline solid (melting point = 127 ° C), purity (HPLC) &gt; 99.8%).

• 9 • 9 • 9

2 °) Method B

Purification was carried out as in Method A, using the mobile phase buffer / methanol (68/32) instead of buffer / tetrahydrofuran.

2 °) Method C

Purification was carried out as in Method A, using the mobile phase buffer / acetonitrile (85/15) instead of buffer / tetrahydrofuran.

Example 28

Preparation of homoharringtonine for pharmaceutical use from semi-purified natural cephalotaxin

The crude homoharringtonine prepared according to Example 25 from partially racemized natural cephalotaxin and purified by chromatography and crystallization according to Method A of Example 27 provides homoharringtonine wherein the content of the unnatural enantiomeric epi-harringtonine is less than 0.05%.

Example 29

Preparation of 2'-de- (methoxycarbonylmethyl) - (2-O-3 ') - dehydroneoharringtonine or cephalotaxylphenylglycidate by esterification of cephalotaxin with phenylglycidic acid

104 ♦ · · * · φ φ φ φ φ

Κ a stirred mixture of (2R, 3R) -cis-phenylglycidic acid (78 mg, 0.48 mmol) in anhydrous toluene (2 mL) was added 1,3-dicyclohexylcarbodiimide (130 mg, 0.63 mmol). The mixture was stirred at room temperature for 10 minutes, and cephalotaxin (50 mg, 0.16 mmol) and pyrrolidine pyridine (24 mg, 0.16 mmol) were added. The mixture was stirred at 18 ± 5 ° C for 15 hours (reaction control C-CM, eluent dichloromethane / methanol; 8: 2), the reaction mixture was filtered on a ground glass filter, the cake was washed with toluene (5 mL) and the filtrate was washed with water. Evaporate in vacuo. The resulting crude product (200 g) was purified by column chromatography (dichloromethane / methanol (98: 2), silica (15-40 µm) 4 g) to give the expected product (19 mg, 27%). The product thus obtained has the following characteristics:

¹ H NMR 400 MHz (CDCl ₃ ) (δ ppm, J Hz):

7.27 (3 H, m, Ph); 7.18 (2 H, m, Ph); 6.63 (1 H, s, H-17 *); 6.40 (1 H, s, H-14 *); 5.96 and 5.85 (2H, 2d, J _AB = 1.5, OCH ₂ O); 5.73 (1H, d, J _3-4 = 9.4, H-3); 5.01 (1 H, s, H-1); 4.01 (1H, d, J ₃ ', _2' = 4.6, H-3 '); 3.65 (3H, s, _OCH3); 3.62 (1H, d, = 9.3, H-4); 3.40 (1H, d, J _{2 '} , ₃ ' = 4.5, H-2 '); 3.27 (1H, m, J _AE = 14.3, J = 12.1 and 7.8, H-11β); 3.05 (1 H, m, H-8a);

2.91 (1H, td, J = 11.7 and 7.4, H-10a); 2.57, m, Η-8β + Η-10β); 2.43 (1H, dd, J _AB = 14.5,

J = 7.0, H-11a); 1.93 (1 H, m, H-6 _A ); 1.84 (1 H, m, H- _6B ); 1.68 (2H, m, CH 2 - 7).

Example 30

Preparation of 2'-de- (methoxycarbonylmethyl) neoharringtonine by hydrogenolysis of cephalotaxylphenylglycidate formed in Example 29

To a stirred solution of cephalotaxylphenylglycidate formed in Example 29 (200 mg, 0.433 mmol) in methanol (10 mL) was added 10% palladium on charcoal (100 mg).

Tf tftf tftf tftf tftf tftf tftf tftf tftf tftf tftf tftf tftf tftf tftf tftf tftf tftf tftf tftf tftf tftf tftf tftf tftf

The resulting mixture was stirred at room temperature under hydrogen pressure (50 psi) for 4 hours and the reaction mixture was filtered and evaporated to dryness. The resulting crude product (175 mg) was purified by column chromatography (dichloromethane / methanol (99: 1 then 98: 2), silica (15-40 µm) 5.5 g) to give an amber solid (86 mg, 43%). ).

The product thus obtained exhibits the following characteristics:

IR (lozenge KBr) (cm ^-1 ). 3436; 2937; 1747; 1655; 1487; 1224 and 1035.

¹ H NMR 400 MHz (CDCl ₃ ) (δ ppm, J Hz):

7.25 (3H, m, mp-Ph); 7.0 (2H, m, o-Ph); 6.65 (1 H, s, H-17 *); 6.63 (1 H, s, H-14); 5.98 (1H, d, J _3,4 = 9.3, H-3); 5.85 (2H, 2d, J _AB = 1.2, OCH ₂ O); 5.09 (1 H, s, H-1); 4.17 (1 H, m, H-2 '); 3.85 (1H, d, J _{4 '} = 9.6, H-4); 3.71 (3H, s, _OCH3); 3.20 (1 H, m, H-11β); 3.10 (1 H, m, H-8a); 2.95 (1 H, m, H-10a); 2.60 (2H, m, Η-8β + Η-10β); 2.39 (2H, m, H-11a + H-3a); 2.04 (1 H, m, H-6 _A ); 2.0 (1H, dd, J _AB = 14.3, J ₃ , _{B. 2} = 9.5, H-3'B); 1.91 (1H, m, H- _6b ); 1.77 (2H, m, _CH2 -7).

Example 31

Preparation of 2'R-de- (methoxycarbonylmethyl) -3'S-azidoneoharringtonine from cephalotaxylphenylglycidate resulting from Example 29

To a stirred solution of cephalotaxylphenylglycidate formed in Example 29 (100 mg, 0.217 mmol) in methanol / water (8/1, 1.27 mL) was added sodium azide (70 mg, 1.08 mmol) and methyl formate (174 µL, 2 µL). , 82 mmol). The mixture was stirred at 50 ° C for 68 hours and cooled to ambient temperature and 5% sodium bicarbonate solution was added to adjust the pH to 8. The resulting aqueous layer was extracted three times with dichloromethane and

106 · · «« * * * '' '' '' '' '' '' '' '' '' '' '' '' '' The combined organic layers were dried over magnesium sulfate and evaporated to dryness.

The resulting crude product (120 mg) was purified by column chromatography (dichloromethane / methanol (99: 1) silica (15-40 µm) 3.5 g) to give a viscous yellow oil (84 mg, 76%). The product thus obtained exhibits the following characteristics:

IR (ATR) (cm ^-1 ): 3488; 2935; 2105; 1748; 1654; 1486; 1223; 1034.

¹ H NMR 400 MHz (CDCl 3) (δ ppm, J Hz):

7.38 (3H, m, m, [beta] -Ph); 7.29 (2H, m, o-Ph); 6.74 (1H, s, H-17 '); 6.67 (1H, s, H-14);

6.08 (1H, d, J ₃ -4 = 9.8, H-3); 5.90 (2H, 2d, J _AB = 1.4, OCH ₂ O); 5.08 (1 H, s, H-1); 4.07 (1H, d large, H-2 '); 3.85 (1H, d, J 4-3 = 9.7, H-4); 3.78 (1H, br. S, H-3 '); 3.69 (3H, s,

OCH3); 3.23 (1 H, m, H-11β); 3.11 (1 H, m, H-8a); 2.98 (1 H, m, H-10a); 2.90 (1 H, d,

J _2' -OH ⁼ 8.2, 2-OH); 2.63 (2H, m, Η-8β + Η-10β); 2.47 (1H, dd, J _AB = 14.2, J = 6.9, H11a); 2.05 (1 H, m, H-6 _A ); 1.92 (1 H, m, H- _6B ); 1.78 (2H, m, _CH2 -7).

Example 32

Preparation of 2 'R-de- (methoxycarbonylmethyl) -3' S-aminoneoharringtonine by hydrogenolysis of the azide resulting from Example 31

To a stirred solution of the product of Example 29 (80 mg, 0.158 mmol) in ethyl acetate-methanol (9/1, 10 mL) was added 10% palladium on charcoal (490 mg). The resulting mixture was stirred at room temperature under hydrogen pressure (50 psi) for 15 hours, and after CCM control, the reaction mixture was filtered and evaporated to dryness.

107 ·· ΦΦ φ φ · · Φ · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · to give a white solid (67 mg, 88%). The crude product thus obtained has the following characteristics:

IR (ATR) (cm ^-1 ): 3299; 2935; 1740; 1654; 1222 and 1034.

¹ H NMR 400 MHz (CDCl 3) (δ ppm, J Hz):

7.27 (5 H, m, Ph); 6.69 (1 H, s, H-17 *); 6.67 (1 H, s, H-14 *); 6.0 (1H, d, J _3-4 = 9.7, H-3); 5.85 (2H, m, OCH ₂ O); 5.09 (1H, br. S, H-1); 4.06 (1H, d, J = 1.2, H-2 '); 3.86 (1H, d, J 4 ₃ = 9.5, H-4); 3.72 (3H, s, _OCH3); 3.38 (1H, br.s); 3.25 (1H, m, Η-11β); 3.14 (1 H, m, H-8a); 2.99 (1 H, m, H-10a); 2.64 (2H, m, Η-8β + Η-10β); 2.49 (1 H, m, H-11a); 2.05 (1H, m, _H-6); 1.94 (1 H, m, H- _6B ); 1.79 (2H, m, _CH2 -7).

Example 33

Preparation of 2'-de- (methoxycarbonylmethyl) -3'-hydroxyneoharringtonine acetonide by esterification of cephalotaxin 1

0.078 mmol) in anhydrous toluene (1 mL) was added 1,3-dicyclohexylcarbodiimide (25 mg, 0.12 mmol). The mixture was stirred at room temperature for 10 minutes and cephalotaxin (75 mg, 0.24 mmol) and pyrrolidine pyridine (12 mg, 0.08 mmol) were added. The mixture was stirred at 18 ± 5 ° C for 15 hours (CCM reaction control, eluent dichloromethane / methanol; 8: 2), the reaction mixture was filtered through a ground glass filter, the cake was washed with toluene (5 mL) and the filtrate was evaporated in vacuo. vacuum. The resulting crude product was purified by column chromatography (dichloromethane, then dichloromethane / methane (98: 2), silica (15-40 µm)) to give the expected product (22 mg, 53%).

i ·· • · · φ · φ φ φ φ φ · φ

108 ¹ Η NMR 400 MHz (CDCl 3) (δ ppm, J Hz):

7.27 (5Η, Ph); 6.63, 6.62, 6.60 and 6.57 (1H, 4s, H-14 *); 6.51, 6.49, 6.42 and 6.41 (1H, 4s, H-17 *); 5.93 (J = 9.6 -4 _3), 5.89, 5.43 (J _4/3 = 9.5) and 5.31 (J _4.3 = 9.3) (1H, 4d, H-3); 5.89 (s), 5.87 + 5.84 (2d, J _AB = 1.5), 5.85 + 5.80 (2d), and 5.84 + 5.77 (2d, J _AB = 1) 5) (2H, OCH 2 O); 5.23 (J = 7.3 -4 _5), 5.20 (J5--4- = 7.4), 4.58 (J5--4- = 8.0) and 4.49 (J ₅ · ^ =

6.2) (1H, 4d, H-5 &apos;); 5.07, 5.03 and 4.83 (1H, 3s, H-1); 4.32 (J ₄ -5 = 7.4), 4.21 (J _{4 -5} = =

6.2), 4.18 (J = ₄ .5- 7.4) and 3.75 (1H, 4d, H-4); 3.86 (J + ₃ = 9.6), 3.76 and 3.60 (1H = 9.5) (1H, 4d, H-4); 3.76, 3.75, 3.70 and 3.43 (3H, 4s, OCH ₃ ); 3.3-1.6 (1H, m); 1.66 + 1.41, 1.65 + 1.37, 1.51 +1.44 and 1.47 + 1.22 (6H, 8s, 2 x CH ₃ )

Example 34

Preparation of cephalotaxyl N-benzyl-3-phenylaziridine-1-carboxylate by esterification of cephalotaxins

To a stirred mixture of (2S, 3S) -cis-N-benzyl-3-phenylaziridine-1-carboxylic acid (360 mg, 1.42 mmol) in anhydrous toluene (5 mL) was added 1,3-dicyclohexylcarbodiimide (390 mg, 1.9 mmol). The mixture was stirred at room temperature for 5 minutes and cephalotaxin (150 mg, 0.47 mmol) and pyrrolidine pyridine (70 mg, 0.47 mmol) were added. The mixture was stirred at 18 ± 5 ° C for 2 hours (CCM reaction control, eluent dichloromethane / methanol; 8: 2), the reaction mixture was filtered on a ground glass filter, the cake was washed with toluene (15 mL) and the filtrate evaporated in vacuo. vacuum. The resulting crude product (785 mg) was purified by column chromatography (dichloromethane / methanol (98: 2), silica (15-40 gm) 23 g) to give a solid (240 mg, 92%). The product thus obtained has the following characteristics:

109 ·· ♦ · 9 · 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 · · 99 ¹ Η NMR 400 MHz (CDCl 3) (δ ppm, J Hz):

7.24 (10H, m, 2xPh); 6.63 (1 H, s, H-17 *); 6.60 (1 H, s, H-14 *); 5.85 and 5.80 (2H, 2d, J _AB = 1.4, OCH ₂ O); 5.64 (1H, d, = 9.3, H-3); 4.97 (1 H, s, H-1); 3.92 and 3.20 (2H, 2d,

Jab = 13.7, CH _? Ph); 3.71 (1H, d, J 4-3 = 9.4, H-4); 3.56 (3H, s, _OCH3); 3.25 (1 H, m, H 11β); 3.07 (1 H, m, H-8a); 2.93 (1 H m, H-10a); 2.86 (1H, d, J ₃ ', _2' = 6.8, H-3 '); 2.57 (2H, m, Η-8β + Η-10β); 2.38 (1H, dd, J _AB = 14.4, J = 7.0, H-11a); 2.07 (1H, d, J ₂ - ₃ =

6.8, H-2); 1.96 (1 H, m, H-6 _A ); 1.82 (1 H, m, H- _6B ); 1.70 (2H, m, _CH2 -7).

Example 35

Preparation of N, O-anhydro-2'-de- (methoxycarbonyl) -3'-benzamidone-harringtonine or cephalotaxyl N, O-anhydro-N-benzoylphenylisoserinate by esterification of cephalotaxin

To a stirred mixture of (4S, 5R) -2,4-diphenyl-4,5-dihydrooxazole-5-carboxylic acid (510 mg, 1.91 mmol) in anhydrous toluene (7 mL) was added 1,3-dicyclohexylcarbodiimide (525). mg, 2.54 mmol). The mixture was stirred at room temperature for 15 minutes and cephalotaxin (200 mg, 0.63 mmol) and pyrrolidine pyridine (95 mg, 0.64 mmol) were added. The mixture was stirred at 18 ± 5 ° C for 3.5 hours (CCM reaction control, eluent dichloromethane / methanol; 9: 1), the reaction mixture was filtered on a ground glass filter, the cake was washed with toluene (15 mL), and the filtrate was washed with water. Evaporate in vacuo. The resulting crude product (1 g) was purified by column chromatography (dichloromethane / methanol (98: 2), silica (15-40 µm) to give a yellow solid (330 mg, 91%). The product thus obtained had the following characteristics:

¹ H NMR 400 MHz (CDCl ₃ ) (δ ppm, J Hz):

·· · »► ^f i ·«

I

110 ’···· ···

8.0 (2H, d, J = 7.3, o-PhC = N); 7.52 (1H, t, J = 7.4, p-PhC = N); 7.44 (2H, t, J = 7.5, mPhC = N); 7.32 (2H, t, J = 7.2, m-Ph); 7.26 (1 H, m, p-Ph); 7.15 (2H, d, J = 7.1, o-Ph); 6.58 (1 H, s, H-17 *); 6.51 (1 H, s, H-14 *); 5.98 (1H, d, J _3-4 = 9.5, H-3); 5.85 and 5.76 (2H, 2d, JAB = 1.3, _OCH2 O); 5.08 (1H, s, H-1); 4.67 (1H, d, J ₄ ', ₅ ' = 5.6, H-4 '); 4.52 (1H, d, J ₅ -4 = 5.6, H-5 '); 3.85 (1H, d, J = 9.6, H-4); 3.70 (3H, s, _OCH3); 3.17 (1H, m, H-11a); 3.08 (1H, m, H-8a); 2.93 (1H, m, H-10a); 2.57 (2H, m, Η-8β + H) -10β), 2.31 (1H, dd, JAB = 14.2, J = 6.8, H-11a); 2.04 (1H, m, H-6 _A), 1.91 (H, m , H-6 _B), 1.75 (2H, m, _CH2 -7).

Example 36

Preparation of N, O-methoxymethylene-2'-de- (methoxycarbonylmethyl) -3'-benzamidoneoharringtonine or cephalotaxyl N, O-methoxymethylene-N-benzoylphenylisoserinate by esterification of cephalotaxin

To a stirred mixture of (4S, 5R) -3-N-benzoyl-2-p-methoxyphenyl-4-phenyloxazolidine-5-carboxylic acid (165 mg, 0.5 mmol) in anhydrous toluene (2 mL) was added 1,3dicyclohexylcarbodiimide (140 mg). , 0.68 mmol). The mixture was stirred at room temperature for 5 minutes and cephalotaxin (53 mg, 0.17 mmol) and pyrrolidine pyridine (25 mg, 0.17 mmol) were added. Stir the mixture at 18 ± 5 ° C for 15 hours (CCM reaction control, eluent dichloromethane / methanol; 9: 1), filter the reaction mixture on a ground glass filter, wash the cake with toluene. (5 ml) and the filtrate evaporated in vacuo. The resulting crude product (230 mg) was purified by column chromatography (dichloromethane / methanol (98: 2), silica (15-40 µm) 7 g) to give the expected product (90 mg, 86%). The product thus obtained has the following characteristics:

111 ¹ H NMR 400 MHz (CDCl ₃ ) (δ ppm, J Hz):

7.66 (2H, br. S, o-BzN); 7.41 (4H, m, BzN + Ph); 7.32 (2 H, m, Ph); 7.26 (2 H, m, Ph); 6.56 (1 H, s, H-17); 6.54 (1 H, s, H-14 *); 5.89 (1H, d, J _4.3 = 9.5, H-3); 5.83 and 5.80 (2H, 2m, OCH 2 O); 5.76 (1H, br. S, H-2 '); 5.10 (1 H, s, H-1); 4.85 (1H, br. S, H-4 '); 4.42 (1H, br. S, H-5 '); 3.84 (1H, d, = 9.5, H-4); 3.72 (3H, s, _OCH3); 3.28 (3H, br. _S , 2'-OCH ₃ );

3.19 (1 H, m, H-11β); 3.09 (1 H, m, H-8a); 2.93 (1 H, m, H-10a); 2.60 (2H, m, Η-8β + Η-10β); 2.37 (1H, dd, J _AB = 14.4, J = 6.6, H-11a); 2.03 (1 H, m, H-6 _A ); 1.90 (1 H, m,

Jab = 12.2, J = 7.8 and 4.4, H-6 _B ); 1.76 (2H, m, _CH2 -7).

Example 37

Preparation of 2'-de- (methoxycarbonylmethyl) -3'-benzamidoneoharringtonine or cephalotaxyl N-benzoyl-phenylisoserinate

1 °) Despite acid hydrolysis of the product of Example 35

To a stirred solution of cephalotaxyl (4S, 5R) -2,4-diphenyl-4,5-dihydrooxazole-5-carboxylate formed in Example 35 (300 mg, 0.53 mmol) in methanol / tetrahydrofuran (50/50 (10 mL)) was added. Add 1N (3.2 mL) hydrochloric acid at room temperature, stir at 18 ± 5 ° C for 3 hours (CCM reaction control), add saturated sodium bicarbonate solution (19 mL) and methanol / tetrahydrofuran 50 / 50 (50 mL) After stirring at 18 ± 5 ° C for 20 hours (CCM reaction control), the reaction mixture was treated with ethyl acetate and water, the resulting aqueous layer was extracted with ethyl acetate, and the combined organic layers were dried over magnesium sulfate and evaporated to a residue. The resulting crude product (170 mg) was purified by column chromatography (dichloromethane / methanol (98: 2), silica (15-40).

112 g 8 g 8 g 8 g 8 g 8 g ) to give a white solid (180 mg, 58%; HPLC purity 92.3%). The product thus obtained has the following characteristics:

[α] _D ²⁰ : -119.2 (c = 0.141; CHCl ₃ ) ¹ H NMR 400 MHz (CDCl ₃ ) (δ ppm, J Hz):

7.75 (2H, d, J = 7.3, o-BzN); 7.51 (1H, t, J = 7.3, [beta] -Bzn); 7.13 (2H, t, J = 7.3, m-BzN);

Ph); 7.27 (5 H, m, Ph); 6.88 (1H, d, J ₃ - _NH = 7.9, 3'-NH); 6.59 (1 H, s, H-17 *); 6.57 (1 H, s, H-14 *); 5.93 (1H, d, J _4.3 = 9.7, H-3); 5.78 and 5.69 (2H, 2d, J _AB = 1.5, OCH ₂ O); 5.06 (1 H, s, H-1); 4.98 (1H, dd, J ₃ , _NH = 7.9, H-3 '); 4.22 (1H, br. S, H-2 '); 3.81 (1H, d, _{J 4 -3} =

9.6, H-4); 3.58 (3H, s, _OCH3); 3.19 (1H, m, J = 12.8, 7.9, H-11β); 3.07 (1 H, m, H-8a);

2.93 (1 H, m, H-10a); 2.72 (1H, br. S, 2'-OH); 2.58 (2H, m, H-8b + H-10b); 2.43 (1H, dd, Jab = 14.2, J = 7.0, H-11a); 2.01 (1H, m, H- _6A ); 1.88 (1H, m, J _AB = 12.0, J = 7.8 and

3.8, H- _6B ); 1.75 (2H, m, _CH2 -7).

2 °) Despite the amidation of the product formed in Example 32

To a stirred solution of the product of Example 32 (60 mg, 0.125 mmol) in ethyl acetate (850 µL) was added saturated sodium bicarbonate solution (850 µL) and benzoyl chloride (19 µL, 0.163 mmol). A white precipitate forms during the reaction. The mixture was stirred at room temperature for 1 hour, then the reaction mixture was diluted with ethyl acetate and the organic layer was washed with saturated sodium bicarbonate solution. The resulting aqueous layer was extracted three times with ethyl acetate and the combined organic layers were dried over magnesium sulfate and evaporated to dryness. The resulting crude product (65 mg) was purified by column chromatography (dichloromethane / methanol (98: 2), silica (15-40 µm) 2.5 g) to give a white solid (41 mg, 56%). The product thus obtained shows the same characteristics as the product obtained in the above method.

Example 38

Preparation of N-tert-butoxycarbamoyl-2'-de- (methoxycarbonylmethyl) -3'-aminoneoharringtonine or cephalotaxyl N-tert-butoxycarbamoylphenylisoserinate via amidation of the product formed in Example 32

113

To a stirred solution of the product of Example 32 (60 mg, 0.125 mmol) in dichloromethane (850 µL) was added saturated sodium bicarbonate solution (850 µL) and dibutyl dicarbonate (27 mg, 0.125 mmol). The mixture was stirred at room temperature for 1 hour, the reaction mixture was diluted with dichloromethane and the organic layer was washed with brine. The resulting aqueous layer was extracted three times with dichloromethane and the combined organic layers were dried over magnesium sulfate and evaporated to dryness. The resulting crude product (70 mg) was purified by column chromatography (dichloromethane / methanol (98: 2), silica (15-40 µm) 3 g) to give a white solid (40 mg, 55%). The product thus obtained exhibits the following characteristics:

¹ H NMR 400 MHz (CDCl ₃ ) (δ ppm, J Hz):

7.27 (3H, m, m, [beta] -Ph); 6.94 (2H, d, J = 6.6, o-Ph); 6.71 (1 H, s, H-17 *); 6.66 (1 H, s, H 14); 6.01 (1H, d, J _4.3 = 9.7, H-3); 5.90 (2H, s, OCH ₂ O); 5.06 (1 H, s, H-1); 5.05 (1 H, m, NH); 4.56 (1 H, m, H-3 '); 4.15 (1 H, m, H-2 '); 3.81 (1H, d, J 4-3 = 9.7, H-4); 3.69 (3H, s, 0CH _3); 3.19 (1H, m, H-11β); 3.10 (1H, m, H-8a); 2.93 (1 H, m, H-10a); 2.61 (2H, m, Η-8β + Η-1Όβ); 2.51 (1H, m, H-11a); 2.05 (1 H, m, H-6 _A ); 1.89 (1H, m, H- _6B ); 1.77 (2H, m, _CH2 -7); 1.44 (9H, s, OC (CH3) _3).

Example 39

Preparation of target. butyl 2-methoxycarbonylmethyl-2-hydroxy-6-methylhept-5-enoate

114 • ···· • · · • and ·· and · 44 4444

94 • · a · 9 4 4

4 4 4

4 4 4 ·· ··

Me / ^ Me / ^QH (ch

2'2

CO ₂ 'But

CH ₂ CO ₂ Me

1 °) Preparation of intermediate oxalate

5-Bromo-2-methylpent-2-ene (1.34 g, 8.2 mmol) was added dropwise to a stirred mixture of magnesium (240 mg, 10 mmol) (activated with an iodine crystal) in anhydrous tetrahydrofuran (8 mL). The course of the reaction is accompanied by a strong overheating and reflux of the reaction mixture. The reflux was maintained until most of the magnesium had reacted and the reaction mixture was diluted with anhydrous tetrahydrofuran (16 mL). To a stirred mixture of tert-butyl ethyl oxalate (1.4 g, 8 mmol) in anhydrous tetrahydrofuran (8 mL) was added the resulting Grignard reagent at -78 ° C over 20 minutes. The temperature was allowed to rise to -15 ° C over 2 hours and the mixture was quenched with 1N hydrochloric acid. The separated organic was washed three times with brine, dried over magnesium sulfate and evaporated to dryness. The resulting crude product (2 g) was purified by column chromatography (cyclohexane / ethyl acetate (98: 2), silica 15 to 40 µη 80 g) to give an oil (660 mg, 39%). The intermediate α-keto ester has the following characteristics:

¹ H NMR 400 MHz (CDCl 3) (δ ppm, J Hz):

5.08 (1 H, m, H-5); 2.80 (2H, t, J = 7.3, _CH2 -3); 2.28 (2H, m, _CH2 -4); 1.68 (3H, s, CH ₃ ); 1.62 (3H, s, CH ₃ ); 1.54 (9H, s, O-t-Bu).

2 °) Preparation of the title compound

To a stirred solution of 1M lithium bis (trimethylsilylamide) in tetrahydrofuran (9 mL, 9 mmol) was added anhydrous methyl acetate (0.7 mL, 8.75 mmol) at -78 ° C over 1 minute and allowed to proceed at -78. ± 5 ° C for 20 minutes. To the stirred mixture

115 • · · · ··· ··· ··· ···· «<

tert-butyl 2-oxo-6-methylhept-5-enoate prepared above (640 mg, 3 mmol) in anhydrous tetrahydrofuran (10 mL) was added lithium enolate at -78 ° C over 5 minutes and the resulting mixture was stirred at -78 ± 5 ° C for 30 minutes. After monitoring in CCM, the freezing bath was removed and the mixture was cooled with 15% ammonium chloride solution (10 mL). The separated organic layer was washed with 15% ammonium chloride solution (10 mL) and evaporated to dryness. The aqueous layer was extracted with ether (2 x 10 mL). The organic layers were combined with the concentrate and washed with brine (10 mL), dried over magnesium sulfate and evaporated to dryness. The resulting crude product (1.3 g) was purified by column chromatography (cyclohexane / ethyl acetate (95: 5), silica (15-40 µm) 60 g) to give an oil (222 mg, 26%). The product thus obtained exhibits the following characteristics:

¹ H NMR 400 MHz (CDCl 3) (δ ppm, J Hz):

5.07 (1 H, m, HC =); 3.67 (3H, s, _OCH3); 3.66 (1 H, s, OH); 2.86 and 2.67 (2H, 2d, J _AB = 15.8 CH 2 CO 2); 2.13 (1H, m, CH ₃ ); 1.85 (1H, m, CH ₃ ); 1.67 (3H, s, CH ₃ ) and (2H, m, CH ₃ ); 1.59 (3H, s, CH ₃ ); 1.51 (9H, s, t-BuO).

Example 40

Preparation of ethyl 2-N - [(4'S) -isopropyl-2'-oxazolidino] carbonylmethyl-2-hydroxy-6-methylhept-5-enoate:

116 • «

Technical solution of 1M lithium bis- (trimethylsilylamide) in tetrahydrofuran (1,28 ml,

1.28 mmol) was added at -78 ° C to a stirred solution of (4S) -3-acetyl-4-isopropyl-2-oxazolidinone (200 mg, 1.17 mmol) in anhydrous tetrahydrofuran and the reaction was allowed to proceed at -78 ° C. for 30 minutes. A solution of ethyl 2-oxo-6-methylhept-5-enoate (323 mg, 1.75 mmol) in anhydrous tetrahydrofuran (5 mL) was then added and the resulting mixture was stirred at -78 ° C for 1 hour. After CCM monitoring, the mixture was cooled with 15% ammonium chloride solution (5 mL). The separated organic layer was washed with 15% ammonium chloride solution (10 mL), then brine (10 mL), dried over magnesium sulfate and evaporated to dryness. The ¹ H NMR spectra of the crude product showed a diastereomeric mixture of about 2.5 / 1. The resulting crude product (516 mg) was purified by column chromatography (cyclohexane / ethyl acetate (90:10 to 80:20), silica (15 to 40 µm) 25 g) to give the minor diastereomer as a yellow oil (55 mg, 13, 7%) showing the following characteristics:

¹ H NMR 400 MHz (CDCl ₃ ) (δ ppm, J Hz):

5.06 (1 H, m, H-3 '); 4.41 (1H, m, H-4 '); 4.25 (4H, m, CH ₂ -5 "and OCH ₂ CH ₃ ); 3.72 (1 H, s, OH); 3.52 and 3.41 (2H, 2d, J _AB 17.9, _CH2 -3); 2.36 (1H, m, H-6 '); 2.16 (1H, m, CH);

1.92 (1H, m, CH); 1.75 (2H, m, CH); 1.67 (3H, s, CH3); 1.59 (3H, s, CH 3); 1.30 (3H, t, J = 7.1, OCH 5 CH 3); 0.89 (3H, d, J = 7.0, CH3); 0.87 (3H, d, J = 6.9, CH3).

The major diastereomer as a yellow oil (93 mg, 23.2%) shows the following characteristics:

¹ H NMR 400 MHz (COCl ₃ ) (δ ppm, J Hz):

5.06 (1H, m, H-3 '); 4.40 (1H, m, H-4 '); 4.23 (4H, m, CH ₂ -5 "and OCH ₂ CH ₃ ); 3.68 (1 H, s, OH); 3.46 (2H, s, _CH2 -3); 2.33 (1H, m, H-6 '); 2.16 (1H, m, CH); 1.91 (1H, m, CH);

1.75 (2H, m, CH); 1.67 (3H, s, CH3); 1.59 (3H, s, CH 3); 1.28 (3H, t, J = 7.1, OCH3 CH2); 0.90 (3H, d, J = 7.0, CH3); 0.87 (3H, d, J = 6.9, CH3).

Example 41

Preparation of ethyl 2- (1'R, 2'S, 5'R) -menthoxycarbonylmethyl-2-hydroxy-6-methylhept-5enoate:

• »

117 • · · ·

99

To a stirred solution of technical 1M lithium bis (trimethylsilylamide) in tetrahydrofuran (14 mL, 14 mmol) was added menthyl (1 R, 2S, 5R) - (-) acetate (2.8 g, 14.1) at -78 ° C. mmol) and allowed to proceed at -78 ° C for 30 minutes.

To a stirred mixture of ethyl 2-oxo-6-methylhept-5-enoate prepared above (867 mg, 4.7 mmol) in anhydrous tetrahydrofuran (12 mL) was added lithium enolate at -78 ° C over 15 minutes and the resulting mixture was stirred at -78 ° C. -78 ° C for 3 minutes (CCM monitoring). The mixture was quenched with 1N hydrochloric acid (30 mL). The separated aqueous layer was extracted with tert-butyl methyl ether (2 x 15 mL) and the combined organic layers were washed with brine (3 x 15 mL), dried over magnesium sulfate and evaporated to dryness. The ¹ H NMR spectra of the crude product showed a diastereomeric mixture of about 6/4. The resulting crude product was purified by column chromatography (cyclohexane / ethyl acetate (98: 2), silica (15-40 µm) 70 g) to give the desired products (1 g, 57%). Separate diastereomers having the following characteristics:

Major diastereomer:

¹ H NMR 400 MHz (CDCl ₃ ) (δ ppm, J Hz):

5.03 (1H, t, J = 7.0, H-3 '); 4.69 (1H, td, J = 10.9 and 4.3, H-1 _Men ); 4.24 (2H, q, J = 7.0, OCH 2 CH 3); 3.77 (1 H, s, 2-OH); 2.91 and 2.67 (2H, 2d, J _AB = 16.4, CH ₂ CO ₂ ); 2.13 (1H, m, CH 1); 1.97 (1H, m, H-6eq _Me ); 1.85 (2H, m, CH 1 and H-7 _Men ); 1.75-1.6 (4H, m, CH ₂ 'and H-3eq, 4EQ _Men); 1.67 (3H, s, CH ₃ '); 1.58 (3H, s, CH ₃ '); 1.45 (1H, m, H-5 _Men ); 1.35 (1H, m, H-2max Men); 1.30 (3H, t, = 6.9, OCH ₂ CH ₃ ); 1.03 (1H, m, H-3 max M _en ); 0.93 (1H, m, H 6ax _Men ); 0.89 (6H, d, J = 6.8, 2 x CH _{3 (Me} n); 0.87 (1H, m, H-4 max M _en ); 0.73 (3H, d, J =

6.9, CH ₃ (Men))

118 φ ·

Minor diastereomer:

¹ H NMR 400 MHz (CDCl ₃ ) (δ ppm, J Hz):

5.05 (1H, t, J = 7.0, H-3); 4.67 (1H, td, J = 10.7 and 4.7, H-1 _M en); 4.25 (2H, m, OCH 2 CH 3); 3.74 (1 H, s, 2-OH); 2.92 and 2.65 (2H, 2d, J _AB = 15.9, CH 2 CO 2); 2.12 (1 H, m, CH); 1.97 (1H, m, H-6eq _M en); 1.86 (2H, m, CH 1 and H-7 _Me n); 1.75-1.6 (4H, m, CH ₂ 'and H-3eq, 4EQ _Men); 1.67 (3H, s, CH ₃ '); 1.58 (3H, s, CH ₃ '); 1.48 (1H, m, H-5 _M- en); 1.36 (1H, m, H-2axMen); 1.31 (3H, t, = 6.9, OOH 7 CH 3); 1.15 -0.8 (3H, m, H-3ax, 6ax, 4ax _Me ); 0.89 (6H, d, J = 6.9, 2 x CH _{3 (Me} n); 0.76 (3H, d, J = 7.0, CH _{3 (Me} n)).

Example 42

Preparation of ethyl 2 - [(R) -1 ', 2', 2'-triphenylethan) -2'-ol] ethoxycarbonylmethyl-2-hydroxy-6-methylhept-5-enoate

A technical solution of 1M lithium bis- (trimethylsilylamide) in tetrahydrofuran (6 mJ, 6 mmol) was added at -78 ° C to a stirred solution of R - (+) - 1,1,2-triphenyl-1,2,2-ethanediol 2-acetate ( 665 mg, 2 mmol) in anhydrous tetrahydrofuran (6 mL). The temperature was then allowed to rise to 0 ° C over 3 hours, and anhydrous heptane (10 mL) was added. To this stirred mixture was added a solution of ethyl 2-oxo-6-methylhept-5-enoate (368 mg, 2 mmol) in anhydrous tetrahydrofuran (2 mL) at -78 ° C and the temperature was allowed to rise to -40 ° over 1 hour. C. After monitoring the CCM, the freezing bath was removed and the mixture was cooled with 15% ammonium chloride solution (10 mL). The separated organic layer was washed with 15% ammonium chloride solution (10 mL) and evaporated to dryness. The aqueous layers were extracted with dichloromethane (2 x 10 mL). The organic layers are combined with the concentrate and

119 ··· · · · «· · · · · · • · · · · · · · · Wash with brine (10 ml), dry over magnesium sulphate and evaporate to dryness.

The ¹ H NMR spectra of the crude product showed a diastereomeric mixture of about 60/40. The resulting crude product (820 mg) was purified by column chromatography (cyclohexane / ethyl acetate (97: 3 to 95: 5), silica (15 to 40 µm) 80 g) to give the desired products (361 mg, 35%). Separate diastereomers obtained as crystalline compounds having the following characteristics:

Minor diastereomer:

¹ H NMR 400 MHz (CDCl 3) (δ ppm, J Hz):

7.66 (2H, d, J = 7.5, o-Ph); 7.43 (2H, t, J = 7.7, m-Ph); 7.35 - 7.0 (11H, m, Ph); 6.72 (1H, s, H-1 '); 4.95 (1H, m, H-3 '), 4.41 (2H, m, _OCH? CH _3); 3.42 (1 H, s, 2-OH); 2,90 a

2.67 (2H, 2d, J _AB = 16.5, CH ₂ -3); 2.53 (1H, s, 2'-OH); 1.98 (1H, m, CH _2); 1.8 to 1.5 (3H, m, CH _2); 1.63 (3H, s, CH ₃ ); 1.52 (3H, s, CH ₃ ); 1.38 (3H, t, J = 7.1, _OCH? CH _3).

Majority diastereomer:

¹ H NMR 400 MHz (CDCl 3) (δ ppm, J Hz):

7.52 (2H, d, J = 7.5, o, Ph); 7.36 (2H, t, J = 7.6, m-Ph); 7.27 (1H, t, J = 7.3, p-Ph); 7.2-7.0 (10 H, m, Ph); 6.59 (1H, s, H-1 '); 4.98 (1H, m, H-3 '); 3.90 and 3.34 (2H, 2m, OCH 2 CH 3); 3.56 (1 H, s, 2-OH); 3.22 (1H, s, 2'-OH); 2.88 and 2.69 (2H, 2d, J _AB = 16.7, CH ₂ -3); 2.06 and 1.79 (2H, 2m, _CH2) 1.7-1.5 (2H, m, _CH2); 1.64 (3H, s, CH _3); 1.54 (3H, s, CH ₃ ); 0.99 (3H, t, J = 7.1, _OCH? CH _3).

Example 43

Preparation of tert-butyl 2 - [(R) -1 ', 2', 2'-triphenylethan) -2'-ol] ethoxycarbonylmethyl-2-hydroxy-6-methylhept-5-enoate

120 ·· ·· · β

9

9 9 9 ·

9 9 9 9 9

9999 98 99

A technical solution of 1M lithium bis- (trimethylsilylamide) in tetrahydrofuran (3 mL, 3 mmol) was added at -78 ° C to a stirred solution of R - (+) - 1,1,2-triphenyl-1,2-ethanediol 2-acetate (330 mg, 1 mmol) in anhydrous tetrahydrofuran (3.5 mL). The temperature was then allowed to rise to -10 ° C over 3 hours, then anhydrous heptane (5 mL) was added. To this stirred reaction mixture was added a solution of tert-butyl 2-oxo-6-methylhept-5enoate (276 mg, 1.5 mmol) in anhydrous tetrahydrofuran (2 mL) at -78 ° C and the temperature was allowed to rise to 1 h over 1 h. —40 ° C. After monitoring the CCM, the freezing bath was removed and the mixture was cooled with 15% ammonium chloride solution (5 mL). The separated organic layer was washed with 15% ammonium chloride solution (5 mL) and evaporated to dryness. The aqueous layers were extracted with dichloromethane (2 x 10 mL). The organic layers were combined with the concentrate and washed with brine (10 mL), dried over magnesium sulfate and evaporated to dryness. The ¹ H NMR spectra of the crude product showed a diastereomeric mixture of about 75/25. The resulting crude product (550 mg) was purified by column chromatography (cyclohexane / ethyl acetate (96: 4, 95: 5 then 90:10), silica (15 to 40 µm) 60 g) to give the desired products (217 mg, 40 mg). %). The major diastereomer thus obtained as a white crystalline compound exhibits the following characteristics:

¹ H NMR 400 MHz (CDCl ₃ ) (δ ppm, J Hz):

7.53 (2H, d, J = 7.4, o-Ph); 7.36 (2H, t, J = 7.6, m-Ph); 7.28 (1H, t, J = 7.3, [beta] -Ph); 7.27.0 (10H, m, Ph); 6.66 (1H, s, H-1 '); 5.00 (1H, m, H-3 '); 3.50 (1 H, s, 2-OH); 2.94 (1H, s, 2'-OH); 2.76 and 2.61 (2H, 2d, J _AB = 16.3, CH 2 -CO 2); 2.06 and 1.78 (2H, 2m, _CH2); 1.65 (3H, s, CH _3); 1.55 (3H, s, CH ₃ and 2H, m, CH ₂ ); 1.23 (9H, s, t-BuO).

Example 44 «·

121

Preparation of 2-carboxymethyl-6,6-dimethyl-2-tetrahydropyrancarboxylic acid or O-demethylanhydrohomoharringtonic acid:

(Method D)

A solution of the ethylene dibasic acid formed in Example 3 (1.5 g, 6.94 mmol) in formic acid (2.6 mL) was stirred at 60 ° C for 16 hours. Thereafter, the temperature of the solution was adjusted to room temperature, the formic acid was removed in vacuo and the resulting crude product was dried at 40 ° C under vacuum for 20 hours (1.5 g, 100%).

Example 45

Preparation of 2-methoxycarbonylmethyl-6,6-dimethyl-2-tetrahydropyrancarboxylic acid or anhydrohomoharringtonic acid from the product obtained in Example 39:

A solution of tert-butyl 2-methoxycarbonylmethyl-2-hydroxy-6-methylheptenoate formed in Example 39 (50 mg, 0.175 mmol) in formic acid (0.5 mL) was stirred at room temperature for 9 days. After removal of the formic acid in vacuo, the residue was treated with 5% sodium bicarbonate solution to pH 8. The aqueous layer was washed with ethyl acetate and then acidified (pH 1) with 1N hydrochloric acid and extracted with ethyl acetate. The organic layer was dried over magnesium sulfate and evaporated to dryness to give anhydrohomoharringtonic acid (20 mg, 50%).

Example 46

122

Preparation of purified (-) cephalotaxin from total alkaloid extract of Cephalotaxus sp

Partially racemized cephalotaxin [H. Wenkui; L. Yulin; P. Xinfu, Scientia Sinica; 23; 7; 835 (1980)] ¹ H NMR of two portions of cephalotaxin (extracted under the same conditions as above) with the optically active NMR sliding agent europium (III) tris [3 (heptafluoropropylhydroxymethylene) - (+) - camphran (1 eq) shows the following results:

Lot A: ¹ H NMR 400 MHz (CDCl ₃ ) (δ ppm): 6.06 (1H, OCH ₂ O (+) - cephalotaxin) and 5.82 (1H, OCH ₂ O (+) - cephalotaxin); 5.99 (1H, OCH ₂ O (-) - cephalotaxin) and 5.76 (1H, OCH ₂ O (-) - cephalotaxin). ·

Presence of 11 ± 5% de (+) - cephalotaxin.

[α] ²² = -134.0 ° (c = 0.214; CHCl ₃ ): calculated ratio of 25 ± 5%

Dose B: slightly racemized (1%) [α] ¹⁹ = -173.3 ° (c = 0.208; CHCl 3)

Enantiomeric enrichment of natural cephalotaxin:

Crude chromatographed cephalotaxin (20 g) was dissolved at 55 ° C in dry methanol (100 mL). Crystallization is effected by cooling on a rotary evaporator and after filtration the product thus obtained shows a purity of 99.9% (HPLC).

[α] D ²⁰ = -130 ° C (C1, CHD ₃ ) corresponds to 10% racemization. The crystalline product thus obtained (20 g) was redissolved in hot methanol (100 ml).

«· • ·

123 *> »

· · ·

Slow cooling of the solution gives a translucent prism composed of pure enantiomeric (-) - cephalotaxin [α] D ²⁰ = -185 ° (C1, CHCl 3) ·

After filtration, the mother liquors are allowed to slowly evaporate at room temperature and the crystals in the form of double needles are exclusively composed of racemic [α] ϋ ^2θ = 0.5 ° (C1; CHCl 3).

After filtration, the second mother liquors provide prisms composed of (-) - cephalotaxin, which is identical to the cephalotaxin obtained in the first crystallization.

After filtration, the mother liquors still provide double needles composed of (±) -cephalotaxin. This cycle is repeated three times. The combined prismatic crystals are recrystallized once to give enantiometrically pure (-) - cephalotaxin, while the coupled double needles treated in the same manner provide 100% racemic cephalotaxin.

Chemical evaluation of enantiomerically pure natural cephalotaxin.

A sample of partially racemized natural cephalotaxin is introduced into the process whose sequence is described in Examples 1,2,3,4,5,6,15,19 and 21 using pure (2R) homoharringtonic acid generated in Example 19. HPLC analysis of diastereomeric mixture the anhydro-homoharringtonine thus obtained shows a significant enantiomeric epihomoharringtonine ratio (11% ± 3%) corresponding to the content of (+) - cephalotaxin in the racemic mixture (it was demonstrated that two antipodes of homoharringtonic acid react in a stoichiometric manner comparable to pure enantiomeric cephalotaxin).

Example 47

Preparation of homoharringtonine from anhydro-homoharringtonine:

• · «·

123 «« 9 '«9

9 ·· 9 »9

• a ··

1 °) A technical solution of hydrobromic acid in acetic acid (17.4 mL, 86.6 mmol, 30% HBr w / w) was added at -10 ° C to a stirred solution of anhydrohomoharringtonine formed in Example 21 (50.8 g, 9 , 63 mmol) in anhydrous dichloromethane. After stirring at -10 ° C for 3 hours, water (240 mL) was added and the reaction mixture became viscous. The temperature was allowed to rise to room temperature, and after stirring for 2.5 hours, sodium bicarbonate (0.76M (406 mL) was added to pH 8. The resulting aqueous layer was saturated with sodium chloride and then extracted with dichloromethane (3 x 230 mL). and the combined organic layers were dried over magnesium sulphate and evaporated to dryness to give a foam After reverse phase chromatography below, 4.03 g of homoharringtonine (77%) was obtained, and the product thus obtained exhibited the same characteristics as in Example 25.

2 °) Method B

To a stirred solution of the anhydrohomoharringtonine formed in Example 21 (214 mg, 0.406 mmol) in anhydrous dichloromethane (1.1 mL) was added a technical solution of hydrobromic acid in acetic acid (0.728 mL, 3.6 mmol, 30% HBr) at -10 ° C. w / w). The mixture was stirred at -10 ° C for 3 hours, water (13 mL) was added and then the temperature was raised to 20 ° C. After stirring at 20 ° C for 3 hours, sodium carbonate solution (0.76M; 31.5 mL) was added to pH 8. The resulting aqueous layer was saturated with sodium chloride, extracted with dichloromethane (3 x 20 mL), and combined. the organic layers were dried over magnesium sulfate and evaporated to dryness. The resulting crude product was purified by reverse phase chromatography below to give homoharringtonine (166 mg, 75%). The product thus obtained shows the same characteristics as the product obtained in Example 25.

125 · '·

Description and definition of harringtonines

CEFALOTAXANY

Examples of harrington acids (R = H), 3

Examples of harringtonines (R = CTX), 4

OH

RO

CEPHALOTAXINES

R ⁴

Examples of cephalotaxins: cephalotaxin = CTXOH

2a

R ¹ = OH, R ² = R 4 = H, R ³ = OMe; acetvlcephalotaxin, 2b

^R1 = Ac, R2 = R4 = H, ^R3 = OMe; cephalotaxinone, 2c

R ^1, R ² = O, R ³ = OMe, R ⁴ is -H:

demethylcephalotaxin

2d

R ¹ = OH, R ² = R ⁴ = H, R ³ = OH;

demethylcephalotaxinone

2e

R ^1, R ² = O. R ³ = OH. R ⁴ = H;

11-hydroxycephalotaxin

2L + 2b = R ⁴ = OH;

2R

2R

CO ₂ H

- after ₂ h

3a citric acid

MeR- (ch ₂ ) 3b co ₂ r

-CO ₂ Me

Harringtonic acid R = H

EXAMPLES OF HARRINGTONQE ACIDS (not isolated in natural form as such)

R ' R ⁷ n R H (CH ₃ ) ₂ C (OH) - 2 H harringtonic acid H (CH ₃ ) ₂ C (OH). 3 H homoharringtonic acid H (CH ₃ ) ₂ C (OH) - 1 H norharringtonic acid H _{(CH3) 2} CH-, 2 H desoxyharringtonic acid -C (CH ₃ ) ₂ -O- 3 H anhydrohomoharringtonic acid · H Ph- 1 H neoharringtonic acid HARRINGTONIN EXAMPLES (natural) H (CH ₃ ) ₂ C (OH) - 2 CTX Harringtonin: HT H (CH ₃ ) _from C (OH). 3 CTX Homoharringtonine: HHT H (CH ₃ ) ₂ C (OH) - 1 CTX Norharringtonin H _{(CH3) 2} CH-, 2 CTX Desoxyharringtonin H _{(CH3) 2} CH-, 1 CTX Nordesoxyharringtonin H _{(CH3) 2} CH-, 3 CTX Homodesoxyharringtonin H _{(CH3) 2} CH-, 4 CTX Bishomodesoxyharringtonin -C (CH ₃ ) ₂ -O- 2 CTX Anhydroharringtonine H Ph- 1 CTX Neoharringtonin H Ph- 2 CTX Homoneoharringtonin

·· • ·

126 • · <

• t · tf · tf

STATE OF THE ART

MeO ₂ C

MeO, C— \ Η ² .Σ> ο

Me- ^ Wn Me β i H) unc OH h ² A ^OH

Me 2 CH ₂ ) _n Me 3 3 HH

Me - ^ 2X10 Me

MeO ₂ C

Face

MeY ^H

Me

H, COOMe

^β ^ ΜθΗ Me

in)

OCTX

4a

(i) impossible due to steric inhibition [K. L. Mikolajczak et al., Tetrahedron, 1995, (1972)]; ii) HCN, H +; iii) MeOH, H &lt; + &gt;; (i) silver salt 3c [K. L. Mikolajczak et al., T Med. Chem., 63, (1975)]; iv) 2a / pyridine-dichloromethane; v) according to [T. R. Kelly et al. Tetrahedron Lett., 3501 (1973)].

Origin of anhydroharringtonines

C. fortunei

Isolation

CO ₂ CTX

CO ₂ Me anhydroharringtonine

Natural homoharringtonine

CO ₂ CTX

CO ₂ Me anhydrohomoharringtonine [DZ Wang et al., Yaoxue Xuebao, p. 173, (1992)] [DZ Wang et al., Yaoxue Xuebao, p. 178, (1992)]

Claims (87)

PATENT CLAIMS A process for the preparation of side-chain cephalotaxan of the following general formula and / or a salt thereof CO-CO-O-CTX characterized in that Ω ("omega") is a representative terminal group of the chain moiety and -CO- is a carbonyl ester group bonded to cephalotaxan; Ω-CO- group corresponds to: - either of the following substituted heterocycloalkane of the general formula: R (CH 2 * n -COwhere n is between 0 and 8; Z is an oxygen, nitrogen or sulfur heteroatom; R ⁵ , R ⁶ and R ⁸ are independently hydrogen; a hydrocarbon group, saturated, unsaturated or aromatic, linear or branched and / or cyclic, in particular alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocycloalkyl, said group including or not including heteroatom (s); R ⁶ and R ⁸ may be incorporated into the cycle; an oxygen ether carrying one of the above groups; - or the following linear alkene of formula: RZH oΛ (t H 2'πι Where m is between 1 and 8, R ⁵ , R ⁶ and R ⁸ are as defined above, - or the following formula wherein n, R ⁵ , R ⁶ and R ⁸ are as defined above; Z and Q ² are independently oxygen, nitrogen or sulfur heteroatoms; Q ¹ is a carbon, silicon, or phosphorus atom; R ⁹ and R ¹⁰ are independently hydrogen, alkoxy, a hydrocarbon group, including or not including heteroarom (s), saturated, unsaturated or aromatic, linear or branched and / or cyclic, especially alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heterocycloalkyl; R ⁹ and / or R ¹⁰ may be zero or together form a heteroatom and / or form a ^1- bond with Q ¹ , R ⁹ and R ¹¹ may be zero or form between two carbon atoms carrying them a multiple bond; and R ¹¹ is hydrogen, arylcarbonyl, alkoxycarbonyl, aryloxycarbonyl or alkyl carbonyl; where -O-CTX is the cephalotaxin moiety of the following formula, a salt thereof: wherein p is 1 or 2; the two types of groups -Ω and -CTX mentioned above are linked by an -CO-O129 ester bond 4 '4 4 4' 4 '4 4' 4 4 '4' 4 '4 4 4 4 44 44 4 4i · 4 · · 4 4 4 4 4 4 4 4 · 4 4 · ··· 44 4 44 · 4 44 4 4 4 4 4 where the procedure includes contacting: either carboxylic acids of the formula Ω-CO-OH or salts thereof; or an active acid form of the formula Ω-CO-A or a salt thereof, wherein Ω-CO has the following general formula R (CH 2'n -COkde n, Z, R ^5, R ⁶ and R ⁸ are as defined above; where Ω-CO has the following general formula: ZH (CH 2'm Ff wherein m is between 1 and 8, Z, R ⁵ , R ⁶ and R ⁸ are as defined above; where Ω-CO has the following general formula: and wherein n, Z, Q ¹ , Q ² , R ⁵ , R ⁶ , R ⁸ , R ⁹ , R ¹⁰ and R ¹¹ are as defined above A means: a cyclic anhydride of the following general formula: O Wherein n, R ⁶ and R ⁸ are as defined above; wherein the reaction is terminated by methylation of the resulting primary carboxyl group with: either cephalotaxan or a salt thereof carrying at least one free hydroxyl group of the formula H-O-CTX, wherein CTX is as defined above; or a metal alkoxide of the formula M-O-CTX, wherein CTX is as defined above and M is a metal; or an active form of its hydroxyl group of the formula YO-CTX, wherein -OCTX is as defined above and Y is either a leaving group to form a negative charge on an oxygen atom by cleavage between Y- and -O-CTX, or allows carbocation by cleavage between YO- and -CTX; optionally in the presence of one or more reaction additives to form a side chain cephalotaxan and / or a salt thereof. 2. A process according to claim 1 wherein Z is an oxygen atom and the cephalotaxan H-O-CTX is cephalotaxin of the following formula or a salt thereof: wherein R ^1, R ^2, R ³ and R ⁴ are independently hydrogen, hydroxyl group or alkoxide. 3. The method of claim 2, wherein said cephalotaxane. HO-CTX, such as cephalotaxin or a salt thereof, wherein R ¹ is hydroxyl, R ² is methoxy, R ³ and R ⁴ are hydrogen. 131 ·: ·::::::::::::::::: A process according to any one of claims 1 to 3, wherein R ⁵ is hydrogen. A process according to any one of claims 1 to 3, wherein R ⁵ is -CH ₂ -CO-O-Me. A process according to any one of claims 1 to 5, wherein n is 1 to 4, R ⁶ and R ⁸ are methyl. 7. The method of any of claims 1-5, characterized mares s c s t s m, n is 1 or 2, ^R6 is phenyl and ^R8 is hydrogen. 8. The method according to claim 1, c h Ujica SET in that R ⁵ is - CH ₂ -CO-O-Me, said cefalotaxan is as stated in claim 3, n = 0, Z is a nitrogen atom and R ⁸ is hydrogen. A method according to any one of claims 1 to 8, wherein A is Ω-CO-O, wherein Ω is as defined in claim 1. A process according to any one of claims 1 to 8, wherein A is a halide. A process according to any one of claims 1 to 8, wherein A is a group of a Ω-CO-A compound having the ability to generate a bond cleavage between a carbonyl group and a substituent A to provide Ω-CO * and AND'. 132 A method according to any one of claims 1 to 8, wherein A is a group selected from substituents: methoxyformyloxy of formula MeOCQO-, trifluoroacetyloxy of formula CF 3 COO-, alkylsulfonyloxy of formula RSO ₂ -, phosphoxy of formula (RO) ₂ POhalophosphoxy of formula ROP (CI) Otrialkylsilyloxy of formula R 3 S 10 -, dimethylformamidium chloride of formula x x. .Me I Cl Me or acyloxypyridinium bromide of formula JAx .A O I Br 'of the formula wherein R is alkyl, The process according to any one of claims 1 to 8, wherein a A is a 2,4,6-trichlorobenzyloxy group 14. A process according to claim 13, wherein the reagent of formula Ω-COA, wherein A is 2,4,6-trichlorobenzyloxy, is obtained by contacting the ΩCO-OH as defined in claim 1. with 2,4,6-trichlorobenzoyl chloride in the presence of one or more O-acylation additives. A process according to any one of claims 1 to 8, wherein the group A corresponds to the following formula: 133 • · · · ······ φ φ · ΦΦΦΦ ·· ♦ '··· φφφ ΦΦΦΦ ΦΦ ΦΦ 16. A process according to claim 15, wherein the component of formula Ω-CO-A is obtained by contacting the acid Ω-CO-CH as defined in claim 1 with carbonyldiimidazole in the presence of strong bases. 17. The method of claim 16, wherein the strong base is an alkoxide. A method according to the preceding claims, characterized in that the coupling additive is a substituted carbodiimide and / or a basic additive, such as a tertiary amine. 19. The method of claim 18 wherein the substituted carbodiimide is selected from cyclohexylcarbodiimide (DCC), 1,3-diisopropylcarbodiimide (DIC), and 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide chlorohydrate. A method according to any one of the preceding claims, wherein the cephalotaxin alkoxide corresponding to the general formula MO-CTX, wherein M and CTX are as defined in claim 1, is prepared by contacting cephalotaxin of the formula HO-CTX, wherein CTX is as defined in of claim 1 with a metal, amide, metal hydride or alkyl metal alone. A process according to any one of claims 1 to 20, wherein M is an alkali metal such as lithium, potassium or sodium. 22: Cephalotaxin lithium alkoxide corresponding to the following formula: 134 ·· · · · · · · · · 23. Sodium cephalotaxin sodium alkoxide corresponding to the following formula: 24. Cephalotaxan with a side chain corresponding to the following general formula and / or a salt thereof: R (CH 2'n -CO-O-CTX wherein n is between 0 and 8; Z is an oxygen, nitrogen or sulfur heteroatom; R ⁵ , R ⁶ and R ⁸ are independently hydrogen; a hydrocarbon group, saturated, unsaturated or aromatic, linear or branched and / or cyclic, in particular alkyl, alkenyl, alkynyl, cycloalkyl. cycloalkenyl, aryl, heterocycloalkyl, said group including or not including heteroatom (s); an oxygenated ether bearing one of the above groups; CTX is as defined in claims 1 to 3 except for compounds wherein Z is an oxygen atom and 1 °) n = 2 or 3 and R ⁶ = R ⁸ = methyl and R ⁵ = OMe or hydroxyl, 2 °) n = 2 and R ⁶ = R ⁸ = methyl and R ⁵ = OMe or hydroxyl, 3 °) n = 3 and at the same time R ⁶ is hydroxyl when R ⁸ is methyl and R ⁵ is CH 2 CO 2 CH 3 135 • «· * '· · 9 9 9 9 9 9 9 25. Side-chain cephalotaxan corresponding to the following formula and / or a salt thereof: R (CH 2 ^H O-CTX) wherein m, R ⁵ , R ⁶ , R ⁸ and CTX are as defined in claim 1 and CTX is as defined in any one of claims 1 to 3 except for the compound where m = 2, R ⁵ CH 2 CO 2 CH 3, R ⁶ = R ⁸ = methyl and CTX is as defined in claim 3. The cephalotaxan of claim 25, wherein R ⁵ is -CH 2 CO-O-CH 3. 27. Side-chain cephalotaxan corresponding to the following formula and / or a salt thereof: wherein n, Z, Q ¹ , Q ² , R ⁵ , R ⁶ , R ⁸ , R ⁹ , R ¹⁰ and R ¹¹ are as defined in claim 1 and CTX is as defined in any one of claims 1 to 3. Cephalotaxan according to claim 27, wherein Q ² is an oxygen atom. 29. The cephalotaxan of claim 28 wherein Z is a nitrogen atom. The cephalotaxan of claim 29, wherein n = 0. 31. Cephalotaxan with a side chain corresponding to the following formula: 136 32. Side chain cephalotaxan corresponding to the following formula: 33. Cephalotaxan with side chain corresponding to the following formula: 137 34. A method according to any one of claims 1 to 21, wherein the cyclic side chain of cephalotaxan and / or a salt thereof of the following general formula: R (CH 2'n -CO-O-CTX wherein n, R ⁵ , R ⁶ , R ⁸ , CTX and Z are as defined in claim 1, said side chain being opened with a reagent and / or a protonic or non-protonic electrophilic radical E in an aqueous or non-aqueous medium to form Intermediate compounds of the following general formula: OE TWt R ⁶ R ⁵ CO ₂ CTX wherein n, CTX, R ⁵ , R ^6, and R ⁸ are as defined above, E is either hydrogen or a provisional or definite electrophile radical; 138 ···· · ·· ·· · · · · φ · φ · φ · Wherein the above-mentioned intermediate compound may be attacked with a reagent or nucleophilic radical Z 'intentionally added or optionally present in the environment, and - if the cyclic side chain of cephalotaxan with the side chain and / or its salt is represented by the following general formula: wherein n, R ⁵ , R ⁶ , R ⁸ , R ⁹ , R ¹⁰ and R ¹¹ are as defined in claim 1 and Z 1 is a heteroatom; said side chain being opened by hydrolysis or by careful solvolysis in the presence of an activating agent or a facilitating agent, optionally; - provides cephalotaxan with an open side chain of the following general formula: Z 'OH - (CH - CO ₂ R ⁶ CTX ^R where n, CTX, R ^5, R ⁶ and R ⁸ are as defined in claim 1; Z'je: - either a halogen or a heteroatom bearing hydrogen or R ¹¹ as defined in claim 1; or hydrogen, a hydrocarbon group wherein the radical bearing or not carrying heteroatom (s) is saturated, unsaturated or aromatic, linear or branched and / or cyclic, especially alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl or heterocycloalkyl. 139 ···· « · * · * * * * * * * «« « 9 · 9 · · ··· The method of claim 34, wherein the cyclic side chain of cephalotaxan with the side chain and / or a salt thereof of the following formula: R (CH 2'n -CO-O-CTX wherein n, R ⁵ , R ⁶ , R ⁸ , CTX and Z are as defined in claim 1, said chain is opened by treatment with a solution of hydrobromic acid in acetic acid in a halogenated solvent, preferably dichloromethane, followed by in situ hydrolysis, to give, without isolation of the intermediate, cephalotaxan with a side chain of the following formula: QH QH R ^CH2> n -CO ₂ CTX where n, CTX, R ^5, R ⁶ and R ⁸ are as defined in claim 1. 36. A process according to any one of claims 1 to 21 and claim 34, wherein the acids are as follows: Ω-CO-OH wherein Ω is as defined in claim 1; wherein said formula is equivalent to a racemic mixture comprising compounds of formulas (+) - O-CO-H and (-) - Ω-ΟΟ-ΟΗ such that (+) - Ω-ΟΟ-ΟΗ represents the dextrorotatory enantiomer and (-) - O-CO-OH is the levorotatory enantiomer, obtained by a) contacting said racemic mixture or one of its active forms of formula CO-CO-A as defined in claim 1; 140 · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · the racemic mixture of said activated form is individually generated by: - either the anion corresponding to formula (Ω-CO-O) '; - or a cation corresponding to the formula (Ω-CO) *; with the pure enantiomeric form of the chiral entity, said "cleavage agent" symbolized by Δ * (delta Stella) has the ability to form: either a stable combination by covalent bonding; or a convertible labile combination by hydrogen bonding or hydrophobic interaction; or a labile intermediate by electrostatic interaction; to provide a diastereomeric mixture of Ω-CO-O-A and Ω-ΟΟ-Δ *, b) then physically separating a mixture of two diastereomers or two complex compounds or more generally two new entities physically and / or chemically different; c) then regenerating and finally separating each enantiomer of the formula ΩCO-OH, where Ω («omega Stella») represents the general symbol of the same chiral radical in either one or other pure enantiomeric forms having the following formulas (+) - Ω-0Ο-ΟΗ; (-) - Ω-ΟΟ-ΟΗ as defined above. 37. The method of claim 36, wherein Ω-CO- is a group corresponding to the following general formula: R (CH ₂ ) n - wherein Z, R ⁶ , R ⁸ and R ⁵ are as defined in claim 1 141 38. The method of claim 36, wherein Ω-CO- is a group corresponding to the following general formula: R (CH3) ZH Wherein Z, R ⁶ , R ⁸ and R ⁵ are as defined in claim 1; 39. The method of claim 36, wherein Ω-CO- is a group corresponding to the following general formula: wherein n, R ⁵ , R ⁶ , R ⁸ , Z, Q ² , Q ¹ , R ⁹ , R ¹⁰ and R ¹¹ are as defined in claim 1. The method of any one of claims 36 to 39, wherein the stable combination is an ester of the following formula Ω-COΟ-Δ, wherein Ω and Δ are as defined in claim 36, wherein the stable combination is obtained by contacting an acid. with a chiral alcohol corresponding to the general formula HOΔ *, where Δ * is as defined in claim 36 according to the process of claim 1. 41. The method of any one of claims 36 to 39, wherein the stable combination is represented by an amide of the following formula Ω-COΝΗ-Δ, or Ω-ΩΟ-Ν-Δ * wherein Ω and Δ * are as defined in claim 36, wherein the stable combination is obtained by contacting the acid with a primary or secondary 142 ····························· A chiral amine corresponding to the general formula Η2Ν-Δ *, or ΝΝ = Δ *, where Δ * has the meaning defined in claim 36 according to the process of claim 1. 42. The method of any one of claims 36 to 39, wherein the stable combination may be represented by a thioester of the following formula Ω-ΟΟ-δ-Δ *, where Ω and Δ * are as defined in of claim 36, wherein the stable combination is obtained by contacting the acid with a chiral thiol corresponding to the general formula Ηδ-Δ *, where Δ * is as defined in claim 36 according to the process of claim 1. The method of any one of claims 36 to 39, wherein the ionic combination may be a salt prepared by contacting an acid with a chiral amine corresponding to one or more of the following three formulas: Ω -CO-O '[NH-Δ] ⁺ Ω -CO-O '[ΝΗ ₂ -Δ *] ⁺ Ω-CO-O '[ΝΗ ₃ -Δ *] ⁺ where Ω and Δ * are as defined in claim 36. A method according to any one of claims 36 to 39, characterized in that an unstable combination is prepared by chiral stationary phase chromatography. 45. The method of any one of claims 36 to 39, wherein the chiral precursor-initiated fractional crystallization is prepared by an interatomic or intermolecular crystalline labile variant. 46. The method of claim 40, wherein the chiral alcohol ΗΟ-Δ * is (-) - quinine corresponding to the following formula: 143 • · · ··· 47. The method of claim 40, wherein the chiral alcohol ΗΟ-Δ * is (-) - or (+) - methylmandelate corresponding to the following formulas: 48. The method of claim 40, wherein the chiral alcohol ΗΟ-Δ * is (-) - or (+) - menthol, corresponding to the following formulas: 49. The method of claim 43, wherein the chiral amine hhN-A * is (-) - or (+) - ephedrine corresponding to the following formulas: 144 • 9 • 9 50. (-) - Quinidyl (2'R) - (-) - anhydrohomoharringtonate and (-) - quinidyl (2'S) - (-) anhydrohomoharringtonate corresponding to the following two formulas: 51. (-) - Menthyl (2'R) - (-) - anhydrohomoharringtonate and (-) - menthyl (2'S) - (-) anhydrohomoharringtonate corresponding to the following two formulas: 52. (-) - Methylmandelyl (2'R) - (-) - anhydrohomoharringtonate and (-) - methylmandelyl (2'S) - (-) - anhydrohomoharringtonate corresponding to the following two formulas: 145 • · · 9 53 (-) - Ephedrine (2'R) - (-) - anhydrohomoharringtonate and (-) - ephedrine (2'S) - (-) anhydrohomoharringtonate corresponding to the following two formulas: 54. The process according to any one of claims 1 to 21 and 34 to 49, wherein the carboxylic acid is: - either a tertiary heterocycloalkanoic carboxylic acid corresponding to the following general formula: R (CH 2'n CO-O-H 146 · '· wherein n, Z, R ^5, R ⁶ and R ⁸ are as defined in claim 1, said acid is obtained by treating in an aprotic solvent optionally in the presence of a cyclizing agent, and / or a dehydrating agent, wherein said treatment is supported by physical removing emerging water. - or an open tertiary ethylenic acid corresponding to the following general formula: Wherein m, Z, R ⁵ , R ⁶ and R ⁸ are as defined in claim 1. - or an open tertiary ethylenic acid corresponding to the following general formula: oh i ^ZH ₂₎ -CO-R ¹² wherein m is 1-8, Z, R ^5, R ⁶ and R ⁸ are as defined in claim 1, R ¹² is not a CTX group as defined in claim 1 and R ⁵ and / or an acid protecting group and / or a chiral group; then R ^{12 is} removed later, either by saponification or hydrogenolysis, or more generally by a method known in the art to remove acid protecting groups. 55. The process of claim 54, wherein the cyclization additive is carried out in the absence of the cyclization additive and the cyclization reaction is performed by heating. 147 · · · · · · · · · · · · · · · · · · 56. The process of claim 54 wherein the cyclizing agent is a protic acid or an aprotic acid included in immobilized form. 57. The process of claim 54, wherein the acid is sulfonic acid or formic acid. A process according to any one of claims 54 to 57, wherein Z is an oxygen atom. 59. Tertiary hydroxycycloalkanoic carboxylic acid including its salts and all racemic mixtures or various mixtures corresponding to the following general formula: (CH 2'n CO-OH wherein n, Z, R ⁵ , R ⁶ and R ⁸ are as defined in claim 1 and R ⁵ is not hydrogen; except for compounds where Z is an oxygen atom and, 1 °) n = 0 and R ⁵ is not -CH 2 CO 2 H or -CH 2 CO 2 CH 3; 2 °) n = 0 and R ⁵ is -CH 2 CO 2 H or -CH 2 CO 2 CH 3 and R ⁶ -R ⁸ = methyl or -CH 2 CO 2 H or -CH 2 CO 2 CH 3; 3 °) n = 2 and simultaneously R ⁶ = R ⁸ = methyl and R ⁵ = OMe or hydroxyl; 4 °) n = 2 and simultaneously R ⁶ = R ⁸ = methyl and R ⁵ is -CH 2 CO 2 H or CH 2 CO 2 CH 3 or methyl; 5 °) n = 3 while R ⁶ is hydroxyl and R ⁸ is methyl and R ⁵ is CH 2 CO 2 CH 3; 6 °) n = 3 and at the same time R ⁶ = R ⁸ = methyl and R ⁵ = OH or methyl or ethyl. 148 • o € · » 9 9 60. Tertiary oxacycloalkanoic carboxylic acid, including its salts and all its pure enantiomeric forms, or in racemic mixtures or in various mixtures, corresponding to the following general formula: R ⁸ ( ^CH 2) n -CO-OH wherein n comprises 0 to 8, R ⁵ , R ⁶ and R ⁸ are as defined in claim 59 but are not simultaneously hydrogen. 61. Tertiary heterocycloalkanoic carboxylic acid, including its salts and all its pure enantiomeric forms, or in racemic mixtures or in various mixtures, corresponding to the following general formula: R ₄ CH ₂ ) -CO-OR wherein n comprises 0 to 8, Z, R ⁵ , R ⁶ and R ⁸ are as defined in claim 59, R ⁵ is not hydrogen and R ¹² is as defined in claim 54. 62. Tertiary oxacycloalkane carboxylic hemiester, including its salts and all its pure enantiomeric forms, or in racemic mixtures or in various mixtures, corresponding to the following general formula: R-O (CH-CO-O-H CH ₂ CO ₂ Me wherein n comprises 0 to 8, R ⁶ and R ⁸ are as defined in claim 59. 149 • · · · 63. Tertiary oxacycloalkane carboxylic hemiester, including its salts and all its pure enantiomeric forms, or in racemic mixtures or in various mixtures, corresponding to the following general formula: R-O • COjR CH ₂ CO ₂ Me wherein n comprises 0 to 8, R ⁶ and R ⁸ are as defined in claim 59, R ¹² is as defined in claim 54. 64. Tertiary oxacycloalkane carboxylic hemiester or anhydro-harringtonic acid, including their salts and all their pure enantiomeric forms, or in racemic mixtures or in various mixtures, corresponding to the following formula: Me Me-CO-O-H CH ₂ CO ₂ Me 65. Tertiary oxacycloalkane carboxylic hemiester or anhydro-harringtonic acid, including their salts and all their pure enantiomeric forms, or in racemic mixtures or in various mixtures, corresponding to the following formula: Me Me-O {CH _2/2 -CO-O-H CH ₂ CO ₂ Me 150 • 9 • 99 9 9 9 9 99 · 9 9 9 99 9 9 9 9 9 9 9 9 9 9 9 99 999998 99 9 999599 99 66. Tertiary oxacycloalkanoic carboxylic acid, including its salts and all its pure enantiomeric forms, or in racemic mixtures or in various mixtures, corresponding to the following general formula: Ph (CH ₂ ) CO-OH wherein n is between 0 and 8 and R ⁵ is as defined in claim 59. 67. Tertiary oxacycloalkanoic carboxylic acid, including its salts and all its pure enantiomeric forms, or in racemic mixtures or in various mixtures, corresponding to the following general formula: Ph (CH 2'n -CO-O-H wherein n is 1 to 8. 68. Tertiary oxacycloalkanoic carboxylic acid, including its salts and all its pure enantiomeric forms, or in racemic mixtures or in various mixtures, corresponding to the following general formula: Ph-O (CH 2'n -CO-O-H CH ₂ CO ₂ Me where n is 0 to 8. 69. Tertiary oxacycloalkanoic carboxylic acid or oxahydroneoharringtonic acid, including their salts and all their pure enantiomeric 151 9 Sep 9 9999 · «9 ·· ·· · 9 9 · 9 9 9 9 9 • · · · · 9 9 9 9 9 • 9 9 9 9 9 9 9 9 9 • 9 9 9 9 9 99 999 · 99 9999 99 99 forms or in racemic mixtures or in different mixtures, corresponding to the following formula: 70. Tertiary oxacycloalkanoic carboxylic acid or oxahydroneoharringtonic acid, including their salts and all their pure enantiomeric forms, or in racemic mixtures or in various mixtures, corresponding to the following formula: Ph-Q ČH: -CO-O-H CH ₂ CO ₂ Me 71. Tertiary oxacycloalkanoic carboxylic acid, including its salts and all its pure enantiomeric forms, or in racemic mixtures or in various mixtures, corresponding to the following formula: PhČH: -CO-O-H 72. Tertiary alkenic carboxylic acid, including its salts and all its pure enantiomeric forms, or in racemic mixtures or in various mixtures, corresponding to the following general formula: ΡΟΗ -CO-O-H ΊΟΗ ₂ ), 2'm Ř 152 wherein m includes between 1 and 8, R ⁶ and R ⁸ are as defined in claim 1, but are not simultaneously hydrogen and R ⁵ is not hydrogen or a heteroatom. 73. Tertiary alkenic carboxylic acid, including its salts and all its pure enantiomeric forms, or in racemic mixtures or in various mixtures, corresponding to the following general formula: Κ ⁰ ΕOH -CO-OH ^(No. ₂ h) ^ _CO2 R wherein m is between 1 and 8, each supporting ring is 1-8, R ⁶ and R ⁸ are as defined in claim 1 and R ¹² is as defined in claim 54th 74. Tertiary alkenic carboxylic acid, including its salts and all its pure enantiomeric forms, or in racemic mixtures or in various mixtures, corresponding to the following general formula: , 8 ROH t ^CH 2) 2'm -CO-O-H Wherein m includes between 1 and 8, R ⁶ and R ⁸ are as defined in claim 1, but are not hydrogen. 75. Tertiary alkenic carboxylic acid, including its salts and all its pure enantiomeric forms, or in racemic mixtures or in various mixtures, corresponding to the following formula: Me MeOH * fCHJ 2'2 -CO-O-H CH ₂ CO ₂ Me 153 ···· · ·> · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · ··· ··· ··· ···· ee ·· 76. Tertiary alkenic carboxylic acid, including its salts and all its pure enantiomeric forms, or in racemic mixtures or in various mixtures, corresponding to the following formula: Me MeOH -CH; -CO-O-H CH ₂ CO ₂ Me 77. Tertiary alkenic carboxylic acid, including its salts and all its pure enantiomeric forms, or in racemic mixtures or in various mixtures, corresponding to the following general formula: w / 7 ^ CH OH 2'm -CO-O-H wherein m is 1 to 8. The compound of claim 77, wherein m = 1. An acid anhydride according to any one of claims 58 to 70 of the formula Ω -CO-O-CO-Ω wherein Ω is as defined in claim 1. A mixed acid anhydride according to any one of claims 58 to 70 of the formula Ω-CO-A, wherein A is a group as defined in any one of claims 12, 13 or 15. 81. Mixed anhydride corresponding to the following formula: 154 • ΦΦ · Φ · ·· φφ ·· φ · · φ · φ · φφφφ · ··· φ · · φ 'φ · φ φφ φ φφφφφφφ Acid chlorides according to any one of claims 58 to 70, corresponding to the general formula Ω-CO-X, wherein X is halogen. 83. A cyclic anhydride corresponding to the following general formula: wherein n, R ⁶ and R ⁸ are as defined in claim 1. 84. A cyclic anhydride corresponding to the following formula: 85. The process of any one of claims 1 to 21 and 34 to 49, wherein the side chain cephalotaxane is purified as a salt by chromatography using a hydrophobic reverse phase as a stationary phase and a mobile phase without an organic solvent such as a solution adjusted to a pH of 2 to 4.5 with a buffer prepared from an acid and a base or an ammonium salt and one or more additives as a silanol effect buffer, wherein said cephalotaxan salt is generated from a mineral acid in a 155 in the form of a chlorohydrate, sulfate, phosphate, perchlorate or organic acid in the form of tartrate, maleate, citrate or lactate. 86. A process according to any one of claims 1 to 21 and 34 to 49, comprising the step of chromatographic purification of natural or semi-synthetic or synthetic homoharringtonine for pharmaceutical use and corresponding to the following formula: to remove undesirable related impurities, namely 2'-epihomoharringtonine, resulting from: (a) either from a semi-synthetic procedure using synthetic homoharringtonic acid of inadequate enantiomeric purity, the impurity generated having an absolute configuration corresponding to the following formula: (b) from a biosynthetic process in plants using cephalotaxin of inadequate enantiomeric purity or in the form of an artifact, by partial racemization of the cephalotaxin moiety, wherein the impurity generated exhibits strictly identical chromatographic properties with a non-chiral system, with an absolute configuration opposite to the above (enantiomer) the following formula: In particular using one of the following chromatographic systems: A) Stationary phase: alkyl- or phenyl- or alkylphenyl- or phenylalkylsilane, preferably n-octadecylsilane, B) Mobile phase: water-tetrahydrofuran, water-methanol, water-acetonitrile or buffer pH 2 to 6.5 to replace water or another mobile phase with equivalent selectivity. 87. A method of purification and chromatographic control according to claim 86 of natural or semisynthetic or synthetic homoharringtonine, which makes it possible to compensate for the double deficiency in the enantiomeric purity of the semi-precursors of both the side chain precursor (said homoharringtonic acid) and cephalotaxin, both said precursors being independently generated by an overall synthetic or semi-synthetic or natural process in the plant (biosynthesis) and, in fact, removal of the unnatural enantiomer of homoharringtonine exhibiting the opposite absolute configuration, using a chiral stationary phase at the production scale. 1/4