BG65893B1 - Hemisynthetic method and new compounds - Google Patents

Abstract

The invention relates to a process for the preparation of a compound with a fused ring structure of the formula which comprises one or more reactions starting from a 21-cyano compound of the formula wherein R 1 is usually an amidomethylene group or an acyloxymethylene group, R 5 and R 8 independently are selected from -H, -OH or -OCOCH2OH, or both R5 and R8 are keto, and ring A is a p-benzoquinone ring, both R14a and R14b are -H ozone is -H and the other is - OH, -OCH3 or -OCH2CH3, or R14a and R14b together form a keto group, and R15 and R18 are independently selected from -H or -OH, or both R5 and R8 are keto, and ring A is a p-benzoquinone ring . In modified starting compounds, the 21-cyano group may be replaced by other groups introduced using nucleophilic reagents.

Description

Technical field

The present invention relates to synthetic methods and, in particular, the present invention relates to semi-synthetic methods.

BACKGROUND OF THE INVENTION

EP 309,477 relates to ecteinascidins 729, 743, 745, 759A, 759B and 770. The ecteinascidin compounds have been described to have antibacterial and other beneficial properties. Ecteinascidin 743 is currently undergoing clinical trials as an antihumor agent.

Ecteinascidin 743 has a complex tris (tetrahydroisoquinoline-phenolic) structure of the following formula (I):

It is obtained by isolation from the extracts of the marine organism Ecteinascidin turbinate. The yield is low and alternative preparative methods are sought.

A synthetic method for the preparation of exgeinascidin compounds is described in US 5,721,362. The method according to this patent is long and complex, there are 38 examples, each of which describes a step of the synthetic sequence until it reaches ecteinascidin 743.

Claim 25 of US 5,721,362 relates to an intermediate phenolic compound of formula (11), which we call Intermediate 11 or Int-11. It has the following bis (tetrahydroisoquinolinophenol) structure (II):

where MOM is a methoxymethyl substituent and TBDPS is a 2, 3H-butyldiphenyl-silyl substituent.

From Intermediate 11 it is possible to synthesize another interesting antitumor agent, phthalacidin, see Proc. Natl. Acad. Sci. USA, 96.3496-3501.1999. Phthalacidin is a bis (tetrahydroisoquinolinophenol) derivative of formula (III):

In ecteinascidin 743, 1,4-bridge there is a structure of formula (IV):

Other known ecteinascidins include compounds with different bridged cyclic ring systems, such as in ecteinescidins 722 and 736, where the bridge is of structure of formula (V):

65893 Bl

ecteinascidins 583 and 597, wherein the bridge is of structure of formula (VI):

ecteinascidins 594 and 596, wherein the bridge is of structure of formula (VII):

The complete structure of these and the corresponding compounds is given in J. Am. Chem. Soc. (1996) 118, 9017-9023. This message is given for comparison.

Other compounds known to be lacking a cyclic ring ring system are known. These include bis (tetrahydroisoquinolinequinone) antitumor-antimicrobial antibiotics safracin and saframycin, and natural seafood renieramycin ihostomycin isolated from cultivated germs or mushrooms. They all have the usual dimeric tetrahydroisoquinoline carbon frame. These compounds can be classified into four types, types I to IV, with respect to oxidation of the aromatic nucleus.

Type I, dimeric isoquinolinquinones, is a system of formula (VIII), and for the most commonly found compounds in this class, see Table I.

Table I

Type I structure of saframycin antibiotics

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Substitutes

Compound _R ^i4a • R ^14b R ² ' _R25a R ^25b _r2 5c saframycin A N. n CN 0 0 CH ₃ saframycin B -H n n 0 0 CH ₃ saframycin C n OCH3 n 0 0 CH ₃ saframycin G n .on CN 0 0 CH3 saframycin H n n CN HE CH ₂ C0CH ₃ CH3 saframycin S n n HE 0 0 CH3 saframycin Y ₃ n n CN NHz H CH3 saframycin Us n n CN NHz H C ₂ H ₅ saframycin A n n CN 0 0 s ₂ n ₅ saframycin Υ # n n CN NHz H N. saframycin Ugh n Q * CN NHz H CH3 saframycin γ ^ n Q * CN NHz H s ₂ n ₅ saframycin AN ₂ n n CN H ^d OH ^e CH3 saframycin AN ₂ Ac n n CN H OAc CH3 saframycin AN, n n CN OH ^a H ^fl CH3 saframycin AN, Ac n n CN OAc H CH3 saframycin AR3 n n N. H OH CH3

Variables are variables ^L kvdeto group Q is of formula (IX):

o ^ u ⁿ CH ₃

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Aromatic rings of type I are given for saframycin A, B and C; G and H; and S is isolated from Streptomyces lavendulae as side compounds. The cyan derivative of saframycin A, called cyanoquinonamine, is known from Japanese Kokai JP-A2 59/225189 and 60/084288. Saframycin Y ₃ , Y _dl , A _dl and Υ _φ are obtained from Streptomyces lavendulae by direct biosynthesis in a suitably selected culture medium. Saframycin Y _2b and Υ _2Μ dimers obtained by binding nitrogen at C-25 from one unit to C-14 from another are also obtained from Streptomyces lavendulae in a suitably selected culture medium. Saframycin ARj (= AN ₂ ), a microbial reduction product of saframycin A at C-25 obtained from Rhodococcus amidophilus, was also obtained by the non-stereoselective chemical reduction of saframycin A with sodium borohydride as an 11 mixture of epimers, followed by chromatography the AC isomer is less polar). The following reducing product saframycin AR ₃ , 21-deciano-25-dihydro-saframycin A, (= 25-dihydrosaframycin B) was obtained by the same microbial conversion. Another type of microbial conversion of saframycin A using Nocardia produces saframycin B, and subsequent reduction with Mycobacterium yields saframycin AS Ac. The 25-O-acetates of saframycin AHj and saframycin AH _{2 were} also chemically obtained for biological testing.

Type I compounds of formula (X) are also isolated from sea mushrooms, see Table II.

Table II

Type I structures of marine mushroom compounds

Substitutes

R143 R14b R ²¹ R renieramycin A HE N. n -C (CH ₃ ) = CH-CH 3 renieramycin B. OS ₂ H ₅ N. n -C (CH ₃ ) = CH-CH 3 renieramycinC he 0 0 -C (CH 3) = CH-CH ₃ renieramycin D OS ₂ H ₅ 0 0 -C (CH3) = CH-CH3 reniramycin E n n he -C (CH ₃ ) = CH-CH 3 renieramycin F OCH3 n he -C (CH ₃ ) = CH-CH 3 xestomycin OCH3 n n -CH3

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Reneramycin A-D was isolated from the antimicrobial extract of a Reniera mushroom collected in Mexico, together with the biogenetically related monomeric isoquinolines rhenirone and the corresponding compounds. The structure of renieri-5 zinc A was initially determined by reverse stereochemistry at C-3, C-11 and C-13. Careful examination of the magnetic resonance data, however, for novel, corresponding renieramycin E and F compounds isolated from the same fungus collected in Palau reveals that the reenamycin ring binding is identical to that of saframycin. This result leads to the conclusion that the previously determined stereochemistry of renieramycin A-D should be the same as that of saframycin.

Xestomycin is found in a mushroom of the species Xestospongia collected from Sri Lanka waters.

Compounds of type II of formula (XI) with reduced hydroquinone ring include saf10 ramicins D and F isolated from Streptomyces lavendulae and saframycin Mx-1 and Mx-2 isolated from Myxococcus xanthus. See Table III.

Table III

Type II compounds

OSN ₃ but ^ A / CH ₃ 0 H 1 I bAan 'T ^ N-r-CH, CH ₃ \ SNZSG ¹ in T nT n R21

Substitutes

Compound R ¹⁴ ' R ^14b R ²¹ _R 3Sa _R 25b safrashcin D 0 0 N. 0 0 saframycin F 0 0 CN 0 0 saframycin MX-1 N. OCH3 HE n CH3 saframycin MX-2 n 0CH3 N. n CH3

R ² *

CH3

CH3 no

NH2

The type III skeleton has been identified in the antibiotics safracin A and B isolated from the ennobled Pseudomonas fluorescens species. These antibiotics of formula (XII) consist of tetrahydroisoquinolinquinone subunit and tetrahydroisoquinolinophenol subunit

65893 B1 where R ²¹ is -H in safracin A and -OH in safracin B.

Saframycin R, the only compound classified as a type IV skeleton, is also isolated from Streptomyces lavendulae. This compound of formula (XIII), consisting of a hydrochinone ring with a glycol ester side chain on one of the phenolic oxygen atoms, is, perhaps, a precursor of saframycin A because of its moderate toxicity.

All these known compounds have a condensed system of five rings (A) to (E), as shown in the following structure of formula (XIV):

Rings A and E are phenolic in ecteinascidins and some other compounds, while in other compounds, especially saframycin, rings A and E are quinone. In known compounds, rings B and D are tetrahydro, while ring C is perhydro.

Object of the invention

The development of new active compounds with the condensed five-ring system of the known compounds is required, as well as alternative synthetic methods for the preparation of ecteinascidin compounds and corresponding compounds. Such synthetic methods can lead to more economical pathways for the production of known antitumor agents, as well as allow the production of new active compounds.

SUMMARY OF THE INVENTION

In one aspect, the present invention relates to the use of a known compound, safracin B, also called quinonamine, in a semi-synthetic method.

Generally, the present invention relates to a semi-synthetic process for the preparation of intermediates, derivatives and corresponding structures of ecteinascidin or other tetrahydroisoquinolinophenol compounds, starting from bis (tetrahydroisoquinoline) alkaloids. Suitable starting materials for the semi-synthetic method include classes of saframycin and safracin antibiotics obtained from different grown broths, as well as classes of reineramycin and xestomycin compounds derived from sea mushrooms.

The general formula (XV) of the starting compounds is as follows:

where:

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R ¹ is an amidomethylene group, such as -CH 2 NH-CO-CR ^25a R ^25b R ^25c , in which R ^25a and R ^25b form a ketone group or one radical is -OH, -NH 2 or -OCOCCH ₃ and the other is -CO ₃ CO 3, -H , -OH or -OCOCH, provided that when R ^25a is OH or -NH 2, R ^25b is not-OH and R ^25c is -H, -CH 3 or -CH ₂ CH ₃ , or R ¹ is an acyloxymethylene group, as -CH-O-CO-R, in which R is -C (CH ₃ ) = CH-CH _{3 or} US;

R ⁵ and R ^{8 are} independently selected from -H, -OH or -OCOCON, or both R ⁵ and R ⁸ are ketogroup and ring A is a p-benzoquinone ring;

both R ^14a and R ^14b are -H or one is -H and the other is -OH, -OCH ₃ or -OCH ₂ H ₃ , or R ^14a and R ^14b together form a ketone group;

R ¹⁵ and R ^{18 are} independently selected from -H or -OH, or both R ⁵ and R ⁸ are ketogroup and ring A is a p-benzoquinone ring; and

R ²¹ is -OH or -CN.

The more general formula of this class of compounds is presented below:

in which each of the substituents defined as R _p Rp R ₃ , R ₄ , R _J , R ₆ , R _? , R ^ R _? , R _J0 e independently selected from the group consisting of H, OH, OCH _3, CN, = 0, OCH _3;

in which X are different amide or ester groups contained in said natural products;

in which each dashed circle represents one, two or three possible double bonds.

Accordingly, according to the present invention, we provide novel semi-synthetic methods for the preparation of intermediates, including intermediate 11 and hence, for the preparation of ecteinascidin compounds, as well as phthalacidin and the corresponding compounds. Each of the semi-synthetic methods of the present invention involves a number of transformation steps to reach the desired product. Each step in itself is a method according to the present invention. The present invention is not limited to exemplary methods which are illustrative, and alternative methods may be presented, for example, by appropriately changing the order of the transformation steps.

In particular, the present invention encompasses the use of a 21-cyan starting material of general formula (XVI):

OCH3 ^R18 \ χ CH3 R5: g

CH3 _x \ X x - 'R15

And -N-CH3

CH ₃ O ^X Υ ^N '' R ^s R1 CN wherein R ^1, R ^5, R ^8, R ¹⁵ and R ¹⁸ are as defined above.

Other compounds of formula (XVI) with different substituents at the 21-position may also be suitable starting materials. In general, any derivative which may be obtained by nucleophilic displacement of the 21-hydroxyl group of the compounds of formula (XV) in which R ²¹ is a hydroxyl group in the cis position. Examples of suitable 21-substituents include, but are not limited to:

mercaptogroup;

an alkylthio group (the alkyl group contains from 1 to 6 carbon atoms);

arylthio group (aryl group contains from 6 to 10 carbon atoms and is unsubstituted or substituted by from 1 to 5 substituents selected from, for example, an alkyl group containing from 1 to 6 carbon atoms, alkoxy groups containing from 1 to 6 carbon atoms , halogen atoms, mercapto groups and nitro groups);

an amino group;

a mono- or dialkylamino group (each alkyl group contains from 1 to 6 carbon atoms), a mono- or diarylamino group (each aryl group is as defined above for the arylthio groups);

65893 B1 is an alpha-carbonylalkyl group of the formula -C (R ^a ) (R ^b ) -C (= O) R ^c , wherein R ^a and R ^b are selected from hydrogen atoms, alkyl groups containing from 1 to 20 carbon atoms, aryl groups (as defined above for the arylthio groups) and aralkyl groups (in which the alkyl group containing from 1 to 4 carbon atoms is replaced by an aryl group defined above for the arylthio group), provided that one of R ^a and R ^b is a hydrogen atom;

R ^c is selected from a hydrogen atom, an alkyl group containing from 1 to 20 carbon atoms, aryl groups (as defined above in the arylthio groups), an aralkyl group (in which an alkyl group containing from 1 to 4 carbon atoms is replaced by an aryl group (defined above for the arylthio group), an alkoxy group containing from 1 to 6 carbon atoms, an amino group or a mono- or dialkylamino group, as defined above.

Therefore, in a general aspect, the present invention relates to methods in which the first step yields a 21-derivative using a nucleophilic reagent. We call these compounds 21-Nuc compounds.

The presence of the 21-cyano group is required for some of the end products, especially ecteinascidin 770 and phthalacidin, while for other end products, it acts as a protecting group that can easily be converted to another substituent, such as the 21-hydroxyl group of ecteinascidin 743 or of 21hydroxyphthalacidin. Using the 21-cyan compound as a starting material effectively stabilizes the molecule during the subsequent synthetic steps until it is separated. Other 21Nuc compounds show these and other advantages.

In one important aspect, the present invention relates to the use of a 21-cyan compound of general formula (XVI) in the preparation of bis- and tris-tetrahydroisoquinolinophenol) compounds. The products that can be obtained include intermediates, such as intermediate 11, and ecteinascidins and phthalacidin, as well as novel and known compounds of the corresponding structure.

Preferred starting materials include those compounds of formula (XV), wherein both R ^14a and R ^14b are hydrogen. Preferred starting materials also include compounds of formula (XV) or (XVI) wherein R ¹⁵ is hydrogen. Moreover, preferred starting materials include compounds of formula (XV) or (XVI), wherein ring E is a phenolic ring. Preferred starting materials also include compounds of formula (XV) or (XVI) wherein at least one, and more preferably, two or three of R ⁵ , R ⁸ , R ¹⁵ and R ¹⁸ are not hydrogen.

Examples of suitable starting materials of the present invention include saframycin A, saframycin B, saframycin C, saframycin G, saframycin H, saframycin S, saframycin Y _J5 saframycin Y _dl , saframycin A _dl , saframycin Y Y, saframycin AN ₂ , saframycin AN ₂ As, saframycin AH _p saframycin AH] Ace, Saframycin AR ₃ , Reneramycin A, Reneramycin B, Reneramycin C, Reneramycin D, Reneramycin E, Reneramycin F, Xestomycin, Saframycin D, Saframycin F, Saframycin MX-2, Saframycin MX-2 , safracin A, safracin B and saframycin R. Preferred starting materials t cyano group in position 21, for the group R ^21.

In a particularly preferred aspect, the present invention includes a semi-synthetic method in which the transformation steps are applied to safracin B:

Safracin B is a ring system very similar to ecteinascidins. This compound has the same pentacyclic structure and the same substituents in the right aromatic nucleus, ring E. Also, safracin B is very close to some of the synthetic intermediates in the general synthesis of ET-743, in particular, to intermediates

65893 Bln. 11. This intermediate can be transformed into ET-743 by using a well known method. Synthetic conversion of safracin B into intermediate 11 therefore results in a semi-synthetic method for the preparation of ET-743.

Therefore, the present invention also relates to intermediate 11 derived from safracin B and to compounds derived from intermediate 11, in particular, ecteinascidin compounds. The present invention also relates to phthalacidin derived from safracin B. The present invention also relates to the use of safracin B in the preparation of intermediate 11, phthalacidin and ecteinascidin compounds, and other intermediates according to the present invention. The present invention also relates to the compounds described herein obtained from other proposed starting materials and the use of these compounds in the preparation of said compounds.

The more preferred starting materials of the present invention have a 21-cyano group. The most preferred compound of the present invention is the compound of formula 2. This compound is obtained directly from safracin B and is considered to be a key intermediate in the semi-synthetic method.

Compound 2

In a related aspect, cyanosafracin B is obtained by fermentation of safracin B obtained from the Pseudomonas fluorescens strain and the culture broth is treated with cyanide ion. The preferred strain of Pseudomonas fluorescens is strain A2-2, FERM BP-14, used in the method according to EP 055, 299. A suitable source of cyanide ion is potassium cyanide. In the usual treatment, the broth is filtered off and an excess of cyanide ion is added. After an appropriate stirring time of 1 h, the pH was left alkaline, 9.5, and extraction with an organic solvent yielded a crude extract, which could then be purified to give cyanosafracin B.

Undoubtedly, the stereochemistry shown in the present description is based on an understanding of the true stereochemistry of natural products. Due to an error in the labeled stereochemistry, an appropriate correction of the formulas presented in this description should be made. Moreover, since the synthesis tends to be modified, the present invention is extended to stereoisomers.

The products of the present invention are generally of formula (XVIIa):

where:

R ¹ is a possibly protected or derivative aminomethylene group, possibly a protected or derivative hydroxymethylene group, with R ¹ being as defined for formula (XV);

R ⁴ eH;

or R ¹ and R ⁴ together form a group of formula (IV), (V), (VI) or (VII):

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R ⁵ is -H or -OH;

R ⁷ is -OCH 3 and R ⁸ is -OH or R ⁷ and R ⁸ together form the group -O-CH 2 -O-; both R ^14a and R ^14b are -H or one is -H and the other is -OH, -OCH ₃ or -OCH ₂ CH ₃ , or R ^14a and R ^14b together form a ketone group; and

R ¹⁵ is -H or -OH;

R ²¹ is -H, -OH or -CN;

and derivatives including their acyl derivatives, in particular where R ⁵ is acetyloxy or another acyloxy group containing up to 4 carbon atoms and including derivatives where the group -NCH ₃ - is replaced by -NH- or -NCH in the 12th position ₂ CH ₃ -, and derivatives in which the -NH ₂ group in the compound of formula (VI) can give derivatives.

In formulas (XVIIa) or (XVIIb), R ¹ is usually aminomethylene, amidomethylene or R ¹ with R ⁴ form a group (IV) or (V). Suitable amidomethylene groups include those of formula -CH2-NH-CO-CHCH3-NH2- derived from alanine and similar groups derived from other amino acids, especially D and L, glycine, valine, leucine, isoleucine, phenylalanine, tyrosine, tryptophan , methionine, cysteine, aspartate, asparagine, glutamic acid, glutamine, lysine, proline, serine, threonine, histidine and hydroxyproline. The general formula for the group R ¹ is -CH 2 -NH-aa, where aa represents an acylamino acid group.

The group R ¹ can be acylated at -NH, the group and, for example, the N-acyl derivatives can be obtained from the groups -CH ₂ NH ₂ and -CH ₁ -NH-aa. Acyl derivatives can be their N-acyl or N-thioacyl derivatives, as well as cyclic amides. The acyl groups may illustratively be alkanoyl, haloalkanoyl, arylalkanoyl, alkenyl, heterocyclylacyl, aroyl, arylaroyl, haloaroyl, nitroaroyl or other acyl groups. The acyl groups may be of the formula -CO-R ^a , where R ^a may be different groups such as alkyl, alkoxy, alkylene, arylalkyl, arylalkylene, amino acid acyl, or heterocyclyl, each of which may be substituted by halo, cyano, nitro , carboxyalkyl, alkoxy, aryl, aryloxy, heterocyclyl, heterocyclyloxy, alkyl, amino or substituted amino. Other acylating agents include isothiocyanates such as aryl isothiocyanates, especially phenylisothiocyanate. The alkyl, alkoxy or alkylene groups of R ^a are suitably containing from 1 to 6 or 12 carbon atoms and may be linear, branched or cyclic. The aryl groups are usually phenyl, biphenyl or naphthyl. The heterocyclic groups may be aromatic or partially or fully unsaturated and may suitably contain from 4 to 8 ring atoms, preferably 5 or 6 ring atoms, one or more of the heteroatoms being selected from nitrogen, sulfur and oxygen.

Without limitation, the R ^a groups typically include alkyl, haloalkyl, alkoxyalkyl, haloalkoxyalkyl, arylalkylene, haloalkylarylalkylene, acyl, haloacyl, arylalkyl, alkenyl and amino acid. For example, R ^a -CO- can be acetyl, trifluoroacetyl, 2,2,2-trichloroethoxycarbonyl, izovalerilkarbonil, trans-3- (trifluoromethyl) cinnamoylcarbonyl, heptafluorbutirilkarbonil, dekanoilkarbonil, transsinamoilkarbonil, butirilkarbonil, 3hlorpropionilkarbonil, cinnamoylcarbonyl, 4metilsinamoilkarbonil, hidrosinamoilkarbonil or transheksenoil- carbonyl or alanyl, arginyl, aspartyl, asparagyl, cystyl, glutamyl, glutaminyl, glycyl, histidyl, hydroxypropyl, isoleucyl, leucyl, lysyl, methionyl, phenylalanyl prolyl, seryl, threonyl, tyronyl, tyronyl ozil, valyl, and other more unusual amino acid acyl groups, as well as phthales11

65893 B1 cluster and other cyclic amides. Other examples can be found in the list of protecting groups.

Compounds in which R ^a -CO- is derived from an amino acid and includes an amino group can form acyl derivatives. Suitable N-acyl compounds include dipeptides that can form N-acyl derivatives.

In one variety; which relates to intermediates, ring A is modified to a structure shown as formula (XX) or (XXI) discussed later.

In another embodiment, which refers to intermediates, the group R ¹ may be -C ₁ H ₂ O-CO-CFu-CH ₂ -S-Prot ³ prepared from a compound of formula (XIX) in which Prot ³ and Fu have the meanings shown. In this case, R ⁷ and R ⁸ form an oxymethylenoxy group. The R ¹⁸ group is usually protected. Typically, R ²¹ is cyano.

R ^14a and R ^14b are preferably hydrogen. R ¹⁵ is preferably hydrogen. O-acyl derivatives are suitable aliphatic O-acyl derivatives containing from 1 to 4 carbon atoms and generally an O-acetal group, especially in the 5th position.

Suitable protecting groups for phenols and hydroxy groups include ethers and esters, such as alkyl, alkoxyalkyl, aryloxyalkyl, alkoxyalkoxyalkyl, alkylsilylalkoxyalkyl, alkylthioalkyl, arylthioalkyl, azidoalkyl, cyanoalkyl, chloroalkyl, heterocyclic, arylacyl, haloarilatsil, cycloalkylalkyl, alkenyl, cycloalkyl, alkylarylalkyl, alkoxy, arylalkyl , nitroarylalkyl, haloarylalkyl, alkylaminocarbonyl-arylalkyl, alkylsulfinylarylalkyl, alkylsilyl and other ethers, and arylacyl, arylalkylcarbonate, aliphatic carbonate, alkylsulfinylarylalkyl-carbonyl m, alkyl carbonate, arylhaloalkyl carbonate, arylalkenyl carbonate, arylcarbamate, alkylphosphinyl, alkylphosphinothioyl, arylphosphinothioyl, arylalkylsulfonate and other esters. It is possible for these groups to be replaced by the above described 1θ groups R ¹ .

Suitable carbamate groups are carbamate, and the like. arylalkenyl, nitroarylalkenyl, heterocyclylalkenyl, heterocyclyl, hydroxyheterocyclyl, alkyldi2 θ thio, alkoxy or halo, or alkylsulfinylarylalkyl, heterocyclylacyl and other carbamates, and alkanoyl Alkalis is a newcomer to the United States of America. groups. It is possible that these 2 θ groups are replaced by the R ¹ groups described above.

Examples of such protecting groups are given in the following tables.

Protection for hydroxyl group

Eteoi Abbreviation methyl methoxymethyl IOM benzyloxymethyl PTO methoxyethoxymethyl MEM 2> (trimethylsilyl) ethoxymethyl SEM methylthiomethyl MTM phenylthiomethyl RTM azidomethyl

65893 B1 cyanomethyl 2,2-dichloro-1,1-difluoroethyl 2-chloroethyl 2-bromethyl tetrahydropyranyl 1-ethoxyethyl phenacyl 4-bromophenacyl cyclopropylmethyl allyl propargyl isopropyl cyclohexyl t-butyl benzyl

2,6-dimethylbenzyl 4-methoxybenzyl o-nitrobenzyl

2,6-Dichlorobenzyl 3,4-dichlorobenzyl 4- (dimethylamino) carbonylbenzyl 4-methylsulfinylbenzyl 9-anthryl methyl

4-picolyl heptafluoro-p-tolyl tetrafluoro-4-pyridyl trimethylsilyl 1-butyldimethylsilyl t-butyldiphenylsilyl triisopropylsilyl

TNT

EE

MRM or PMB

Msib

TMS

TBDMS

TBDPS

TIPS

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Esters arylformate aryl acetate arylevulinate arylpivaloate ArOPv arylbenzoate aryl-9-fluorocarboxylate arylmethyl carbonate 1-adamantyl carbonate t-butyl carbonate BOC-OAG 4-methylsulfinylbenzylcarbonate Msz-Oar 2,4-dimethylpent-3-yl carbonate Doc-Oar aryl-2,2,2-trichloroethyl carbonate aryl vinyl carbonate arylbenzylcarbonate arylcarbamate dimethylphosphinyl Dmp-OAr dimethylphosphinothioyl Mpt-OAr diphenylphosphinothioyl Dpt-Oar arylmethanesulfonate aryltoluenesulfonate aryl-2-formylbenzenesulfonate Amino group protection Carbamate Abbreviation methyl ethyl 9-fluorenyl methyl Fmoc

9- (2-sulfo) fluorenylmethyl

9- (2,7-dibromo) fluorenylmethyl

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17-tetrabenzo [a, c, e, |] fluorenylmethyl Tbfmoc 2-chloro-3-indenylmethyl Climoc benzShinden-3-ylmethyl Bimoc 2,7-di4-butyl [9- (10,1O-dioxo-10,10,10,10- tetrahydrothioxanthyl) methyl DBD-Tmoc 2,2,2-trichloroethyl Troc 2-trimethylsilylethal Teoc 2-phenylethyl hZ 1- (1-adamantyl) -1-methylethyl Adpoc 2-chloroethyl 1,1-dimethyl-2-chloroetal 1,1-dimethyl-2-bromoethyl 1,1-dimethyl-2,2-dibromethyl DB-t-BOC 1,1-dimethyl-2,2,2-trichloroethyl TCBOC 1-methyl-1- (4-biphenyl) ethyl Bpoc 1- (3,5-di-1-butylphenyl) -1 -1-methylethyl t-Burmeoc 2- (2'-and 4'-pyridyl) ethanol Pyoc 2,2-bis (4'-nitrophenyl) ethyl Bnpeoc n- (2-pivaloylamino) -1,1-dimethylethyl 2 - [(2-nitrophenyl) diethyl] -1-phenylethyl NpSSPeoc 2- (n, n-Dicyclohexylcarboxamido) ethyl t-piston BOC 1-adamantyl 1-Adoc 2-adamantal 2-Adoc vinyl Voc allyl Aloe or Alloc 1-isopropylalyl Ipaoc synamyl Coe 4-nitrosinamyl Noah 3- (3'-pyridyl) prop-2-enyl Paloc

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8- quinolyl n-hydroxypiperidinyl alkyldithio benzyl p-methoxybenzyl p-nitrobenzyl p-bromobenzyl p-chlorobenzyl

2,4-Dichlorobenzyl 4-methylsulfinylbenzyl

9-Antryl methyl diphenylmethyl phenothiazinyl- (10) -carbonyl p-p-toluenesulfonylaminocarbonyl n'-phenylaminothiocarbonyl Amides formamide acetamide chloracetamide trifluoroacetamide phenylacetamide 3-phenylpropanamide

3- pyridyl carboxamide benzamide p-phenylbenzamide p-phthalimide p-tetrachlorophthalimide

4- nitro-p-phthalimide

Cbz or 3

Moz

PNB

Msz

TFA

TCP

65893 Bl n-Dithiasuccinimide p-2,3-diphenylmaleimide p-2,5-dimethylpyrrole p-2,5-bis (triisopropylsiloxyl) pyrrole p-1,1,4,4-tetramethyldisilyazacyclopentane aduet

1,1 D-Tetramethyl-1,3-disillaisoindoline Special -NH protected groups p-methylamine nt-butylamine p-allylamine n- [2- (trimethylsilyl) ethoxy] methylamine n-3-acetoxypropylamine p-cyanomethylamine n- (1- isopropyl-4-nitro-2-oxo-3-pyrrolin-3-yl) amine n-2,4-dimethoxybenzylamine

2-Azanorbornene n-2,4-dinitrophenylamine n-benzylamine n-4-methoxybenzylamine n-2,4-dimethoxybenzylamine n-2-hydroxybenzylamine n- (diphenylmethyl) amino p-bis (4-methoxyphenyl) methylamine n-5-dibenzosubersamine p-triphenylmethylamine p-C4-methoxyphenyl) diphenylmethyl] amino n-9-phenylflurenylamine p-ferrocenylmethylamine

Dts

BIPSOP

STABASE

BSB

SEM

Dmb

Bn

MRM

DMPM

Hbn

DPM

DBS

Tr

MMTr

Pf

Fem

65893 Bl n-2-picolylamine n'-oxide n-1,1-dimethylamometallenamine n-benzylidenamine n-p-methoxybenzylidenamine n-diphenylmethyleneamine p- (5,5-dimethyl-3-oxo-1-cyclohexenyl) amine n-nitroamine n-nitrosoamine diphenylphosphinamide Dpp dimethylthiophosphinamide Mpt diphenylthiophosphinamide Ppt dibenzylphosphoramidate

2-nitrobenzenesulfenamide Nps n-1- (2,2,2-trifluoro-1,1-diphenyl) ethylsulfenamide TDE

3- nitro-2-pyridinesulfenamide Npys p-toluenesulfonamide Ts benzenesulfonamide

Safracin B includes an alanyl side chain. In one aspect, the present invention relates to the protection of a free amino group with a Boc group, which has great advantages.

Some ecteinascidines of the present invention include compounds of formula (XVIII):

65893 B1 especially group (IV) or (V);

R ²¹ represents hydrogen, a hydroxyl group or -CN, especially a hydroxyl group or CN;

and derivatives thereof, in particular, 5-acyl derivatives including a 5-acetyl derivative.

Preparation of ecteinascidin 743 and related compounds

In general, conversion of the 21-cyan starting compound to an ecteinascidin product, for example, of formula (XVIII) includes:

(a) if necessary, conversion of the quinone system of ring E into a phenolic system;

(b) if necessary, conversion of the quinone system of ring A into a phenolic system;

c) conversion of the phenolic system of ring A into the methylenedioxy-phenol ring;

d) forming a bridged spiro ring system of formula (IV), (VI) or (VII) through a 1-position and a 4-position in ring B; and

e) suitable transformation such as acylation.

Step a) if necessary, the conversion of the quinone ring system E to the phenolic system can be carried out by conventional reduction methods. A suitable reagent is hydrogen with a palladium-on-carbon catalyst, but other reducing systems may be used.

Step b), if necessary, conversion of the quinone system of ring A into a phenolic system is analogous to Step a) and no further details are needed.

Step c) conversion of the phenolic system of Ring A into a methylenedioxyphenol ring can be carried out in several ways, possibly with Step b). For example, the quinone ring A may be demethylated at the 7-position methoxy substituent and reduced to dihydroquinone and treated with a suitable electrophilic reagent, such as dibromomethane, bromochloromethane or similar bivalent reagents, which directly produces the methylenedioxy ring system, or with a divalent ring system, or with a divalent such as thiocarbonyldiimidazole when producing a substituted methylenedioxy ring system which can be converted to the desired ring.

Step d) usually proceeds by appropriately replacing the 1st position with a bridging reagent that can assist in the formation of the desired bridge, obtaining the exo-quinone method of the 4th position and allowing the method to interact with the 1-substituent to obtain a bridging structure. Preferred bridge reagents have the formula (XIX)

PgG Fu where Fu is a protected functional group, such as -NHProt ^4a or OProt ⁴ ® groups, Prot ³ is a protecting group and the dotted line indicates a possible double bond.

The method is prepared by first inserting a hydroxyl group at the 10-position at the bond between rings A and B to give a partial structure of formula (XX):

or, preferably, a partial structure of formula (XX):

wherein the group R 'is selected for the desired group of formulas (IV), (V), (VI) or (VII). For the first two of these groups, R 'represents -CHFuCH ₂ -SProt ₃ . The protecting groups can then be separated and suitably modified to give the desired compound.

A typical method for Step d) is shown in US 5,721,362 for comparison. In particular, the passage in column 8, step (1) and Example 33 of the patent.

The modification in Step e) may include acylation, for example with the Ra-CO- group, as well as conversion of the 12-NCH ₃ group to 12-NH or

65893 Bl

12-NCHjCHj. Such conversion may be carried out before or after the other steps, by appropriate methods.

By way of illustration, it is possible to transform cyanosafracin B of formula 2 into ecte-5 insascidine ET-743, which is a shorter and more accurate method than previously used. Cyanosafracin B can be transformed into intermediate 25 and from this compound it is possible to obtain a number of cysteine derivatives, which can be further transformed into ET-743. Preferred cysteine derivatives are shown as compounds Int-29 and Int-37.

lnt-29

Int-37

The retrosynthetic analysis for the preparation of ET-743 using compound 29 is shown in Scheme I.

NHTroc

ET-743

INT-27

Scheme I

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According to Scheme I above, it is possible to obtain ET-743 in 21 linear steps. In this method, cyanosafracin B is transformed into intermediate 25 after a series of interactions involving mainly (1) the separation of a methoxy group present in ring A, (2) the reduction of ring A and the formation of methylenedioxy group in a single vessel, (3) hydrolysis of the amide functionality found at carbon 1, (4) converting the resulting amino group into a hydroxyl group. This method also achieves the protection and deprotection of primary alcohol functionality at position 1 of ring B of compound 25 by directly using cysteine residue 29 to afford intermediate 27. Cysteine derivative 29 is protected at the amino group with beta, beta , a beta-trichloroethoxycarbonyl protecting group, to obtain compatibility with existing allyl and MOM groups. Intermediate 27 is directly oxidized and cyclized. These circumstances, as well as the different strategy of deprotection in the later stages of synthesis, make the method new and more susceptible to industrial development than the method described in US 5,721,362.

The conversion of the 2-cyan compound to intermediate 25 usually involves the following steps (see Scheme P):

preparing a protected compound of formula 14 by reacting 2 with tert-butoxycarbonyl anhydride;

conversion of 14 to a disubstituted compound of formula 15 by reaction with bromomethyl ether and diisopropylethylamine in acetonitrile;

selective elimination of the methoxy group of the quinone system in 15 to obtain a compound of formula 16 by reaction with methanol sodium hydroxide solution;

transforming 16 into a methylenedioxy compound of formula 18 in the following preferred sequence: (1) the quinone group of compound 16 is reduced by 10% palladium on carbon in a hydrogen atmosphere; (2) the hydroquinone intermediate is converted to the methylenedioxy compound of formula 17 by reaction with bromo-chloromethane and casein carbonate in an atmosphere of hydrogen; (3) 17 is transformed into a compound of formula by protecting the free hydroxyl group as the OCH ₂ R group. The reaction is carried out with BrCH ₂ R and casein carbonate, where R may be aryl, CH = CH 2, OR 'and the like.

eliminating the tert-butoxycarbonyl and methyloxymethyl protecting groups of 18 to afford a compound of formula 19 by reaction with a solution of hydrogen chloride in dioxane. This reaction is also carried out by mixing 18 with a solution of trifluoroacetic acid in dichloromethane;

preparing the thiourea compound of formula 20 by reaction with phenylisothiocyanate;

converting a compound of formula 20 into an amine compound of formula 21 by reaction with a solution of hydrogen chloride in dioxane;

transformation of compound 21 into a NTgos derivative 22 by reaction with trichloroethyl chloroformate and pyridine;

preparing a protected hydroxy compound of formula 23 by reacting 22 with bromomethyl ether and diisopropylethylamine;

transformation of compound 23 into an N-H derivative 24 by reaction with acetic acid and zinc;

conversion of the compound of formula 24 into hydroxy compound 25 by reaction with sodium nitrite in acetic acid. Alternatively, nitric tetroxide may be used in a mixture of acetic acid and acetonitrile, followed by treatment with sodium hydroxide. Sodium nitrite may also be used in a mixture of acetic anhydrous acetic acid, followed by treatment with sodium hydroxide.

65893 Bl

Conversion of intermediate 25 to ET-743 using cysteine derivative 29 is generally performed in the following steps (see Scheme III): 45 transformation of a compound of formula 24 into derivative 30 by protecting the primary hydroxyl functionality with (S) -N- 2, 2, 2-trichloroethoxycarbonyl-8- (9H-fluoro-9-ylmethyl) cysteine29; 50

Scheme II conversion of the protected compound of formula 30 into phenolic derivative 31 by cleavage of the allyl group with tributyltin hydride and dichloropalladium bis (triphenylphosphine);

transformation of the phenolic compound of formula 31 into a compound of formula 32 by oxidation with benzene selenic anhydride at low temperature;

65893 B1 transformation of the hydroxyl compound of formula 32 into lactone 33 in the following sequence: (1) reacting a compound of formula 32 with 2 eq triflic anhydride and 5 eq dimethyl sulfoxide; (2) reacts with 8 eq diisopropylethylamine; (3) reacts with 4 eq t-butyl alcohol; (4) reacts with 7 eq 2-1-butyl-1, 1, 3, 3 tetramethylguanidine; (5) reacts with 10 eq acetic anhydride;

transforming lactone 33 into hydroxyl compound 34 by removing the MOM protecting group with TMSI;

cleavage of the N-trichloroethoxycarbonyl group of the compound of formula 34 to compound 35 by reaction with zinc / acetic acid;

transformation of the amine compound into the corresponding alpha-ketolactone 36 by reaction with N-methylpyridinecarboxaldehyde chloride followed by DBU;

preparing ET-770 by reacting a compound of formula 36 with 3hydroxy-4-methoxyphenylethylamine;

transformation of ET-770 into ET-743 by reacting with silver nitrate in an AcN / water mixture.

Et-770

AJNOj

CH3SNIGO

Et-743

Scheme III

65893 Bl

The above-described method for the transformation of intermediate 25 into ET-743 can be efficiently modified using other cysteine derivatives, for example, compound 37, named 2-methoxymethyloxy-3- (9H-fluorene-9-ylmethyl) thiopropene acid. This compound already has a ketone group in the form of enol ether, while in the other cysteine analogues there is an amino group that must later be transformed into a ketone group in a transamination reaction with a median yield of 55-60%. Therefore, a significant increase in linear fusion yield is possible using compound 37, since the transamination step is missed.

The conversion of an intermediate to ET-743 using cysteine derivative 37 can be carried out in the same manner and with the same reagents unlike cysteine derivative 29 in the absence of transformations (e) and (g). The reaction sequence is presented in Scheme IV:

39

Et-770 Et-743

65893 Bl

Compound 38 can also be prepared by reacting intermediate 12 described in US 5,721, 362 with intermediate 37 in improving the method described in this patent.

Preparation of phthalacidin and related compounds

According to the present invention, a class of compounds include phthalacidin and have the following general formula (XX):

OSN ₃

wherein R ¹ is an amidomethylene group; R ⁵ is a short oxy-side chain; and R ²¹ is a cyano group or a hydroxyl group. For phthalacidin, R ¹ is a phthalimidomethylene group; R ⁵ is an acetoxy group; and R ²¹ is a cyano group. The other groups for R ¹ include mono- and di-N-substituted amidomethylenes, as well as other cyclic amidomethylenes, and the other groups for R ⁵ include more C 1 -C ₄ acyl groups as well as C 1 -C ₄ alkyl groups.

Conversion of the 21-cyan compound to phthalasidine or the corresponding compound of formula (XX) typically involves the following steps:

(a) if necessary, conversion of the quinone ring system E into a phenolic system;

b) forming a -R ⁵ group at the 5 position of ring A;

c) forming an R ¹ group at the 1-position 5 of ring B; and

(d) if necessary, conversion of the quinone ring system A into a phenolic system;

e) conversion of the phenolic ring system A into the methylenedioxy-phenol ring.

These steps are closely similar to the steps shown for the preparation of ecteinascidins. Stage

c) usually involves the formation of the CH ₂ NH _{2 group} in the first position and its acylation.

Phthalacidin can be prepared by 15 using the intermediates described in the conversion of cyanosafracin B to intermediate 25. For example, intermediates 21 and 17 are suitable starting materials for the preparation of phthalacidin.

As shown below in Scheme V, the method for the synthetic preparation of phthalacidin from intermediate 21 involves the following steps:

transforming 21 into a compound of formula 25 by reaction with phthalic anhydride in dichloromethane and carbonyl diimidazole;

conversion of 27 to phthalacidin by reaction with tributyltin hydrochloride and dichloropalladium-bis (triphenylphosphine) or basic medium followed by interaction with acetyl chloride.

Scheme V

65893 Bl

As shown in Scheme VI, the method for the synthetic preparation of phthalacidin from intermediate 17 involves the following steps:

acetylation of the hydroxyl group of a compound of formula 17 with acetyl chloride and pyridine to give an acetylated intermediate of formula 42;

separation of the t-butoxycarbonyl and methyloxymethyl protecting group 42 to afford a compound of formula 43 by reaction with a solution of hydrogen chloride in dioxane. This reaction can also be carried out by mixing 42 with a solution of fluoroacetic acid in dichloromethane;

preparing a thiourea compound of formula 44 by reacting 43 with phenyl 5 isothiocyanate;

converting the compound of formula 44 into an amine compound of formula 45 by reaction with a solution of hydrogen chloride in dioxane;

The transformation of 45 into phthalicidin by reaction with phthalic anhydride in dichloromethane and carbonyl diimidazole.

PiwnmgHnqiarHt CHjCh

phthalicidin

Scheme VI

Preparation of intermediate 11 and the corresponding intermediates Retrosynthetic analysis is described in the following sequences:

Safracin B

65893 Bl

In the present invention, a key class of intermediates include intermediate 11 and have the general formula (XXI):

OSN ₃

I

PgO3 ^^ A ^ / CH3

OProt ² wherein Prot ¹ and Prot ² are hydroxyl protecting groups, preferably different. Typically, Prot ¹ is selected from [more generalisation needed]. Typically, Prot ² is selected from [more generalisation needed]. For intermediate 11 itself, the Prot ¹ group is a methoxymethyl group and Prot ² is a t-butyldiphenylsilyl group.

Conversion of the 21-cyano compound to intermediate 11 or the corresponding intermediate of formula (XXI) typically involves the following steps:

(a) if necessary, conversion of the quinone ring system E into a phenolic system;

b) forming -Oprot ¹ group at the 18th position of ring E;

c) forming a CH ₂ -OProt ² group at the 1-position of ring B; and

(d) if necessary, conversion of the quinone ring system A into a phenolic system;

(e) conversion of the phenolic ring system A into a methylenedioxyphenol ring.

Step (b), formation of the -OProt ¹ group at the 18th position of ring E, is a simple reaction of protecting a phenolic group and does not need to be explained. Appropriate conditions are chosen depending on the nature of the protecting group. The other steps are the same as the other reactions.

Step (c), formation of the CH ₂ -OProt ² group in the Eva position of ring B, is usually carried out by first obtaining the CH ₂ NH _{2 group} in the first position and then converting the amino group into a hydroxyl group and protecting. Therefore, when the starting compound has a group R ¹ which is -CH 2 NHCO-CR ^25a R ^25b R ^25c , then the Nacyl group must be separated. When the parent compound has a group R ¹ which is -CH 2 -O-CO-R, then no changes in ecteinascidin are required, where the substituent R ¹ is the same. For other products, the O-acyl group must be separated. This deacylation can be accomplished by various methods. In one, the deacylation and conversion to hydroxyl functionality are carried out in one step. The hydroxyl group may then be acylated or otherwise modified to give the appropriate group R ¹ .

US 5,721,362 describes synthetic methods for the preparation of ET-743 by long, multi-step synthesis. One of the intermediates in this synthesis is intermediate 11. Using cyanosafracin B as a starting material, it is possible to obtain intermediate 11 in a much shorter way to obtain such an intermediate and, therefore, to improve the process for preparing ET-743.

Cyanosafracin B can be converted to intermediate 25 by the methods described above. From intermediate 25 it is possible to obtain intermediate 11 using the following steps, see Scheme VII:

preparing a protected hydroxyl compound of formula 26 by reacting 25 with tert-butyldiphenylsilyl chloride in the presence of a base;

final cleavage of the allyl group with tributyltin hydride and dichloropalladiumbis (triphenylphosphine) at 26, resulting in intermediate 11.

65893 Bl

Scheme VII

One aspect of the synthetic method of the present invention for transforming safracin B into intermediate 11 is modifying and extending Scheme VIII and considering the following steps:

stereospecific conversion of the compound of formula 1 (safracin B) into the compound of formula 2 by selective substitution of a hydroxyl group with a cyano group by reaction with potassium cyanide in an acidic medium;

preparing the thiourea compound of formula 3 by reacting the compound of formula 2 with phenylisothiocyanate;

conversion of the thiourea compound of formula 3 into acetamide of formula 5 under hydrolysis in an acidic medium followed by addition of acetic anhydride. The intermediate amine of formula 4 can be isolated by fixing the hydrolysis in acidic sodium bicarbonate medium, but this intermediate is very unstable and quickly turns into a five membered cyclic imine, called compound 6;

preparing a protected compound of formula 7 by reaction with bromomethylmethyl ether and diisopropylethylamine in dichloromethane;

selective demethylation of the quatoin system's methoxy group of the compound of formula 7 in a compound of formula 8 by reaction with methanol sodium hydroxide solution;

converting the compound of formula 8 into the methylenedioxy compound of formula 9 according to the following preferred steps: (1) the quinone group of compound 8 is reduced by 10% palladium on carbon in a hydrogen atmosphere;

(2) the hydroquinone intermediate is converted to the methylenedioxy compound of formula 9 by reaction with bromochloromethane and cesium carbonate in an atmosphere of hydrogen;

(3) the compound of formula 9 is transformed into a compound of formula 10 by protecting the free hydroxyl group as an OCH ₂ R group. The reaction is carried out with BrCH ₂ R and cesium carbonate, where R may be aryl, CH = CH 2, OR'ht.h .;

converting the acetamide group of the compound of formula 10 into the corresponding hydroxyl group of the compound of formula 11 by reaction with nitric tetraoxide in a mixture of acetic acid and acetyl acetate, followed by treatment with sodium hydroxide;

or alternatively, sodium nitrite may be used in a mixture of acetic anhydride and acetic acid, followed by treatment with sodium hydroxide;

or alternatively, the acetamide group of the compound of formula 10 may be converted to a primary amino group by reaction with hydrazine or with Boc ₂ O, DMAP followed by hydrazine;

such a primary amine may be converted to the corresponding hydroxyl group (compound of formula 11) by oxidative conversion of the primary amine to the corresponding aldehyde with

65893 Bl

4-formyl-1-methylpyridinium benzenesulfonate or other pyridinium ion, followed by treatment with DBU or other base and subsequent hydrolysis and then reduction of the aldehyde to the corresponding hydroxyl group with lithium aluminum hydride or other reducing agent;

preparing a protected compound of formula 26 by reaction with t-butyldiphenylsilyl chloride and dimethylaminopyridine in dichloromethane;

transformation of the silylated compound of formula 26 into intermediate 11 by deprotecting the OCH ₂ R protecting group by reacting under reducing or acidic conditions.

Palladium black is generally used in an atmosphere of hydrogen or aqueous trifluoroacetic acid, or tributyltin hydride and dichlorobis (triphenylphosphine palladium).

In another preferred modification, the cyan compound of formula 2 can be transformed into intermediate 11 using Extended Scheme II, comprising the steps of:

preparing a protected hydroxyl compound of formula 26 by reacting 25 with tert-butyldiphenylsilyl chloride in the presence of a base;

final cleavage of the allyl group with tributyl tin hydride and dichloropalladiumbis (triphenylphosphine) at 26, resulting in intermediate 11.

Preparation of the active compounds

It is possible to transform cyanosafracin B into a number of intermediates and derivatives with potential antitumor therapeutic activity. These intermediates can be prepared from compounds already described or using alternative methods.

The intermediates described in the present invention include compound 47 and a number of amide derivatives obtained using compounds 45 or 43.

Scheme VIII illustrates the preparation of compound 47 using the following sequence:

preparing the thiourea compound of formula 3 by reacting the compound of formula 2 with phenylisothiocyanate;

conversion of the thiourea compound of formula 3 into acetamide of formula 5 under hydrolysis in acidic medium followed by addition of acetic anhydride. The intermediate amine of formula 4 can be isolated by fixing the hydrolysis in acidic sodium bicarbonate medium, but this intermediate is very unstable and quickly turns into a five membered cyclic imine, called compound 6;

preparing a protected compound of formula 7 by reaction with bromomethylmethyl ether and diisopropylethylamine in dichloromethane;

selective demethylation of the methoxy group of the quinone system of the compound of formula 7 in a compound of formula 8 by reaction with methanol sodium hydroxide solution;

converting the compound of formula 8 into the methylenedioxy compound of formula 9 according to the following preferred steps: (1) the quinone group of compound 8 is reduced by 10% palladium on carbon in a hydrogen atmosphere; (2) the hydroquinone intermediate is converted to the methylenedioxy compound of formula 9 by reaction with bromochloromethane and cesium carbonate in an atmosphere of hydrogen; (3) the compound of formula 9 is transformed into a compound of formula 10 by protecting the free hydroxyl group as an allyloxy group by reaction with allyl bromide and cesium carbonate;

transforming the compound of formula 9 into acetyl derivative 46 by reaction with acetyl chloride in pyridine;

transformation of the compound of formula 46 into unprotected compound 47 by reaction with hydrochloric acid in dioxane.

65893 Bl

Scheme VIII

Other useful derivatives of the amide intermediates are obtained from the already described intermediate 45 using the following scheme. The second stage is possible. The method is an important part of the present invention, especially when the group R is R ^a , as defined above. Even more,

Scheme VIII can be easily extended to indicate the preparation of the compounds of formula (XXIII) upon introduction of the starting material with a different group at the 5-position, or a group that remains directly in the product, or a group that may be separated or modified to obtain the desired group.

RCOCI. ru

Scheme IX

65893 Bl

From compound 45, a group of analogues can be obtained according to the following scheme:

acylating the amino group of compound of formula 45 with a number of acyl derivatives to provide the corresponding amides, as a preferable 5 Chita acyl groups are acetyl, cinnamoyl chloride, p-trifluorsinamoilhlorid, izovalerilhlorid phenyl isothiocyanate or amino acids or give contractors above examples of the groups R ^a CO-; transformation of the cyano group into a hydroxyl group by reacting with silver nitrate in an AcN / water mixture.

Other useful derivatives of amide intermediates are obtained from the already described intermediate 43 using the following scheme:

RCOC1. ru

Scheme X

From compound 43, another group of interesting derivatives can be obtained using the following scheme:

(a) acylating the amino group of compound of formula 43 with a number of acyl derivatives to provide the corresponding amides, as preferred acyl groups are acetyl, cinnamoyl chloride, p-trifluorsinamoilhlorid, izovalerilhlorid or amino acids or denier above examples of the groups R ^a CO -;

(b) transformation of the cyano group into a hydroxyl group by reaction with silver nitrate in an AcN / water mixture.

New intermediates

In the light of the above explanations, it can be seen that the present invention relates to novel intermediates. Depending on Ring A, the intermediates are of formula (XXPa):

65893 Bl

where:

R ¹ is -CHANEL, or CH ₂ OH, or a protected or derivative variant of this group and R ⁴ is -H; or R ¹ and R ⁴ together form a group of formula (IV), (VI) or (VII):

R ⁵ is a hydroxyl group or a protected or derivative variant thereof; both R ^14a and R ^14b are hydrogen or one is hydrogen and the other is hydrogen or a protected or derivative of this group, -OCH3 or OCH2CH3, or both R ^14a and R ^14b together form a ketone group; R ¹² is hydrogen, -CH 3 or -CH ₂ CH ₃ ; R ¹⁵ is a hydroxyl group or a protected or derivative thereof; R ¹⁸ is a hydroxyl group or a protected or derivative thereof; and R ²¹ is -H, -OH or -CN.

In one aspect of the present invention it is preferred that at least one of the groups R ¹ , R ⁵ , R ^14a , R ^14b , R ¹⁵ or R ¹⁸ be a protected or a derivative group.

In one aspect of the present invention, the group R ¹ is not a 3, N-butyldiphenylsilyl substituent and / or the group R ¹ is not a methoxymethyl group.

Preferably R ¹ is -CH ₂ NH ₂ or CH ₂ OH or a protected or derivative of this group and R ⁴ is -H; or R ¹ and R ⁴ together form the group:

Both R ^14a and R ^14b are preferably hydrogen.

One preferred class of intermediates according to the present invention includes the compound we identified as compound 25 of the formula:

65893 Bl

Therefore, the preferred class is of the general formula in which the IOM group is replaced by some other protecting group.

Other preferred intermediates according to the present invention include the compound we identified as compound 45 and 47. Other N-acyl derivatives can be readily prepared from compound 45 and are an important part of the present invention. Suitable acyl groups include the acyl groups described above. The corresponding 21-hydroxyl compound is also useful and finds its place among the active compounds obtained by us.

New active compounds

We have also found that some of the compounds of the present invention, which were originally obtained as intermediates, have exceptional activity in the treatment of cancer, such as leukemia, lung cancer, colon cancer, kidney cancer and melanoma.

Accordingly, the present invention relates to a method of treating mammals, especially a human, suffering from cancer, comprising administering to a diseased individual a therapeutically effective amount of a compound of the present invention, or a pharmaceutical composition thereof contains.

The present invention also relates to pharmaceutical compositions containing as active ingredient a compound or compounds of the present invention, and methods for their preparation.

Examples of pharmaceutical compositions include solids in the form of tablets, pills, capsules, granules, etc., or a liquid (solutions, suspensions or emulsions) of a suitable composition for oral, topical or parenteral administration, which may contain pure a compound or a mixture thereof with a carrier or other pharmacologically active compounds. These formulations need to be sterile when administered parenterally.

The administration of the compounds or compositions of the present invention can be accomplished by any suitable method such as intravenous infusion, oral compositions, intraperitoneal and intravenous administration. The infusion time is preferably up to 24 hours, more preferably 2 to 12 hours, and most preferably 2 to 6 hours. Short infusion times that allow treatment to be carried out without a hospital stay are particularly desirable. However, if the infusion may be required to last from 12 to 24 hours or longer, the infusion may be administered at suitable intervals of 2 to 4 weeks, the pharmaceutical compositions containing the compounds of the present invention may be delivered by liposomal or nanospheric encapsulation, in restraint compositions or by other standard means.

The dose of the compounds will vary depending on the individual composition, the route of administration and the individual situs, host, and tumor to be treated. Other factors such as age, body weight, gender, diet, administration time, rate of discharge, patient's condition, drug combination, response sensitivity, and severity of the disease are also considered. Administration may be carried out continuously or periodically with the maximum tolerated dose.

The compounds and compositions of the present invention may be used with other medicines for combination therapy. The other drugs may be part of the same composition or administered as a separate composition at the same time or at another time. The nature of the other medicine is not limited, with suitable ones including:

(a) antimitotic drugs, especially those intended for cytoskeletal elements, including microtubule modulators such as taxane drugs (such as taxol, paclitaxel, taxotere, dosetaxel), podophyllotoxins or vinca alkaloids (vincristine, vinblastine);

b) antimetabolite medicines such as 5fluorouracil, cytarabine, gemcitabine, purine analogues (such as pentostatin, methotrexate);

c) alkylating agents such as nitrogen mustards (such as cyclophosphamide or ifosfamide);

(d) drugs intended for deoxyribonucleic acid, such as anthracycline drugs adriauicin, doxorubicin, pharmorubicin or epirubicin;

65893 Bl

(e) medicines for the use of zatopoisomerases, such as etoposide;

f) hormones and hormonal agonists and antagonists such as etherogens, antiestrogens (tamoxifen and related compounds) and androgens, flutamide, leuprorelin, goserelin, cyprotrone or octreotide;

(g) drugs intended for signal transduction in tumor cells, including antibody derivatives such as herceptan;

h) alkylating agents such as platinum drugs (cis-platinum, carboplatin, oxaliplatin, paraplatin) or nitrosocarbamides;

i) drugs that potentially act on tumor metastases, such as matrix cell kletoproteinase inhibitors;

(j) gene therapy agents;

(k) antibody therapists;

(l) other bio-active compounds of marine origin, especially didemnins such as aplidine;

m) steroid analogues, in particular dexamethasone;

n) anti-inflammatory agents, in particular dexamethasone; and

o) vomiting medicines, in particular dexamethasone.

The present invention also relates to the use of the compounds of the present invention in the method of treatment and to the use of the compounds in the preparation of a pharmaceutical composition for the treatment of cancer.

Cytotoxic activity

Cell cultures. The cells were kept in logarithmic growth phase in Eagle's Minimum Essential Medium, with Earle's Balanced Salts, with 2.0 mM L-glutamine, with basic amino acids, without sodium bicarbonate (EMEM / pea); supplemented with 10% fetal calf serum (FCS), ΙΟ ' ² M sodium bicarbonate and 0.1 g / l penicillin-G + streptomycin sulfate.

A simple screening procedure was performed to determine and compare the antitumor activity of these compounds using the adapted form of the method described by Bergeron et al (1984). The tumor cells used were P388 (DBA / 2 mice lymphoid neoplasm suspension culture), A-549 (human lung carcinoma monolayer), HT-29 (human colon carcinoma monolayer) and MEL-28 (monolayer culture). from human melanoma).

P-388 cells were seeded in 16 mm wells in an amount of 10 x ⁴ cells per well in 1 ml aliquots of MEM 5FCS containing the drug at the indicated concentration. Non-drug cultures are planted in a separate set to monitor control growth and to determine whether cells remain in the exponential phase of growth. All determinations are made twice. After three days of incubation at 37 ° C, under an atmosphere of 10% carbon dioxide in 98% moisture, an estimated IC ₅₀ was determined to compare growth in drug wells with that in control wells.

A-544, HT-29 and MEL-28 cells were seeded in 16 mm wells in 1 x 10 ⁴ well cells in 1 ml aliquots of MEM 5FCS containing the drug at the indicated concentration. Non-drug cultures are planted in a separate set to monitor control growth and to determine whether cells remain in the exponential phase of growth. All determinations are made twice. After three days of incubation at 37 ° C, under a atmosphere of 10% carbon dioxide in 98% moisture, the wells were stained with 0.1% Crystal Violet. The approximate IC _{50 was determined} to compare growth in drug wells with that in control wells.

1. Raymond J. Bergeron, Paul E Cavanaugh, Jr., Steven J. Kline, Robert G. Hughes, Jr., Gary T. Elliot, Carl W. Porter. Antineopiastia and antiherpetic activity of spermidine catecholamide iron chelators. Biochem. Bioph. Really. Comm. 1984, 121 (3). 848-854.

2. Alan C. Schroeder, Robert G. Hughes, Jr., Alexander Btoch. Effects of Acyclic Pyrimidine Nucleoside Analoges. J. Med. Chem. 1981, 24, 1078-1083.

It can be seen from the data on cytotoxic activity and other perceptions that the active compounds of the present invention include a preferred class of compounds of general formula (XXIII):

where:

R ¹ is as defined for formula (XVIIb) and is preferably a medium-sized aminomethylene group;

65893 Bl

R ⁵ is as defined for formula (XVIIb) and is preferably a small size hydroxyl group;

R ¹² is as defined above and is preferably -CH ₃ and

R ²¹ is hydroxyl or cyano.

R ¹ is a suitable hydrophobic group and not free amino, hydroxyl or other hydrophilic functionalities. Typically, R ¹ is the group -CH ₂ -NH-CO-R 1 in which R ^a is as defined, but preferably has a linear chain of less than 20 atoms, more preferably less than 15 or 10 atoms, with 1,4-phenyl being counted as a chain of 4 to 5 atoms, and this applies to other cyclic groups (for example, 1,2-cyclohexyl is counted as a chain of 2 atoms) and the linear chain itself of less than 10,15 or 20 atoms may be substituted. In particular, the data show that a balance is achieved between the non-existent

1C (CM) P-388 Α-549 ΗΤ-29 MEL-28 CV-1 DU-145 OM * NH 0.009 0.018 0.018 0.018 « 0.023 cm · • & Mm 0.15 > 0.15 0.15 > 0.15 1? "* 15 1.44 1.44 1.44 1.44 j OM · '16 > 1.5 > 1.5 > 1.5 > 1.5 * \ -0 X See O 1 A π 1.4 1.4 1.4 1.4 X 1 "· ^1-0 V 1 ο ** ⁴ * '_' 18 0.01 0.01 0.01 0.01 . * 19 0.08 0.16 0.01 0.16

65893 Bl

° U l ^ b. ⁰ x 20 0.01 0.01 0.01 0.01 x “A * ^Vo 21 0.019 0.019 0.019 . 0.019 • a. oh · X “A \ Ws- You I L oA ^ ojCft 22 0.014 0.014 0.014 0.014 0.014 0.014 4? “H * Υι ^ζ ΥΓ * ' ^ι 3 * ¹¹ · Vo X. cm Nh ₀ A _e , ov =% 23 0.13 0.13 0.13 0.13 0.13 0.13 % ° * “ζ _ο / -. Χγ“ <Y'Z 24 0.18 1.8 1.8 1.8 1.8 1.8 > 1 A 1 fly » '' - “Υ. 25 0.2 0.2 02 0.2 0.2 press L · OM · ° * t ⁱⁿ 1 Aβθ \ L JTJ ** Ύ ^ 'χιφ ¹ * vq in * 35 0.008 0.008 0.008 0.008 Q E OM »° * Ύ * ή ^ΗΟ ν ^ 4 · ^'11 · *? K / JCX Vo Έν 36 0.01 0.01 0.01 0.01 In J £ n Υ W 28 0.001 0.001 0.001 · 0.001 0.001 0.001 1? * Q * £ J2 1 xC ⁰ \ 42 0.13 0.13 0.13 0.13 0.13

65893 Bl

OM · those jLI V-o ZjbN 43 0.008 0.016 0.008 0.008 0.016 ' on ^^^ m · Ote J7 T ^ YYY ^ t * * L ₀ Ι ^ δΝ ⁰ '44 0.001 0.001 0.001 0.001 0.001 ΟΜ »OAC jlT 'OTQ. ₄₅ 0.01 0.01 0.01 0.01 0.01 CM · “λ ^- β / ^γ * ^ι * Ύ ^χ ^ or CN NK K« SC <yA> _o 0.015 0.015 0.015 0.015 0.018 < 2.171 2.171 2.171 2.171 2.171 cv * mv * yy * t ⁰ s> A 5 0.005 0.005 0.005 0.005 1 ^ M · o ^ Q- _t ^ L _r Mt w JL Ifi M-1J ° 4h «o * <7 0.22 022 0.22 0.22 0.22 '? "· ^Ο · ^ · ^Ο > | ^ γ" · * ϊΫΥτ * AA -'- Z ^e k. * <A 8 > 9 > 18.1 > 18.1 > 18.1 > 18.1

65893 Bl

1 “· oh IJ Me. Us. -4 · *. V ^ r ^ * rw · ** · * ⁴ · oW>'ήη 9 > 1.77 > 1.77 > 1.77 > 1.77 > 1.77 \ _o o _v o ^ ru ζΧΧΧ ^ v _e oA 10 > 1.65 > 1.65 > 1.65 > 1.65 « > 1.65  j GAM «VUy“ · - & hStccf A « 0.016 0.016 0.016 0.016 0.016 Hmm · OAC Ie w oA 47 0.001 0.001 0.001 0.001 0.001 OM · Do it. 48 0.0008 0.001 0.0008 0.0008 0.001 OM » o * e T J lib Axy ^ T jOOO ** ° Vo I cm No 49 0.007 0.007 0.007 0.007 0.007 ohm *. MO ^ J ^ M · oi jrj m «A -. Τ ^ ΊΓ * · '> “· JL · / ^Vo 4> ΆΓ _Μ 0.0001 0.0001 0.0001 0.0001 0.0001 ohms νο ^ Χ, μ · . UDF ΊΓ J o ¥ r '/ VO CN ΜΗ οΑ * “ 51 <51 0.0001 0.0001 0.0001 0.0001 0.0001 οαμ ΟΑβ j M · ^ .Us. ίΥ ^ τ * 1 “· o'SAr '? ^ 4o & N MH 0-S * Ml h> 52 0.001 0.001 0.001 0.001 0.001

65893 Bl

owe OAC J2 j ftte -X. j **. γΥτ * 7 * Λγζ • 0 \ CH ₀ ^ γσ, me 0 53 0.0001 0.0001 0.0001 0.0001 0.0001 ' % = “* + N><-ο \ Sn _e JyNH _Y ”, me β 54 0.001 0.001 0.001 0.001 0.001 ome he \ J \ »0 \ & N NH Ο ^ γΝΗγ. », Me 0 55 0.01 0.01 0.01 0.01 0.01 QUt + r S6 0.18 0.9 0.18 0.8 0.9 Cue 57 0.14 0.14 0.14 0.14 0.14 Oita o ^/ vS ' ^, *' ^szl * '' ^u . * '• * T ~ Gy 0.001 0.001 0.001 0.001 0.001 «E Μ? * Ύγν5 £ me ο Μ 60 0.001 0.001 0.0005 0.001 0.0005 I "" X + '"t'SI _β ^ζ ' Τ ^Α Τ ^Μ 'Τ' -Ο 'CN ΗΗ« Αν, 6ΐ 0.001 0.001 0.001 0.001 0.001 ο * 6 ΊΓ | _o jpu> ν.ο χ On NH _u ^OD X “T ^ 62 0.001 0.001 0.0005 0.0005 0.001

65893 Bl

OM · OM! j ο lnn V 63 0.0001 • · · · · · · 0.0001 ~ —Rr-- · ··· ···· 0.0001 ·· '··· 0.0001 0.0001 · OMe HCs ^ Ym · OM j D NM w - »· ° 64 0.001 0.001 • 0.001 0.001 0.001 CM * CM ° \ -0 1 CH 65 0.0001 0.0005 0.0001 0.0001 0.0005 QMe MO ^ hhMe GM IT j Vo (OH 0 66 0.0001 0.0001 0.0001 0.0001 0.0001 du · ohm 17 JT V-O \ OH №1 * I 67 0.0001 0.0001 0.0001 0.0001 0.0001

the group R ^a -CO- and having a large group.

In one aspect of the present invention, it is preferred that in R ¹ there are no cyclic groups, especially aromatic groups. In the same sense, the present invention does not refer to the preparation of the compounds described in Proc. Natl. Acad. Sci. USA 96,3496-3501,1999 given for comparison. The R ¹ groups we prefer exclude the corresponding CH ₂ R ₂ substituents shown in Table 1 of the above message, especially the groups A, B, CnDsaRj.

R ⁵ is preferably an acetyl group.

In the particularly preferred compounds of the present invention, the R ¹ group is acylated on the -NH ₂ group, for example, N-acyl derivatives can be obtained from the groups -CH ₂ NH ₂ and -CH ₂ NH-aa. The acyl derivatives can be their N-acyl or N-thio acyl derivatives. The acyl groups may be of the formula -CO-R ^a in which R ^a is as defined above and is selected to satisfy the criteria shown. Suitable acyl groups include alanyl, arginyl, aspartyl, asparagyl, cystyl, glutamyl, glutaminyl, glycyl, histidyl, hydroxypropyl, isoleucyl, leucyl, lysyl, methionyl, phenylalanyl, propyl, seryl, threyl, threyl, threyl, threyl, threyl, threyl, threyl, threyl, threyl, threyl, threyl, threyl other acyl groups of amino acids. Such acyl groups of amino acids are preferably formed on the amino group to give hydrophobicity.

In one aspect of the present invention, the R ¹ group is a hydroxy methylene derivative. It may also be a derivative of an aminomethylene group.

An important method of the present invention in which the active compounds are prepared is the following:

65893 Bl

where R ⁵ for the final product is as defined for compound (ΧΧΠΙ) and may be different in the starting material and altered during the process,

R ¹⁸ is a hydroxyl group in the final product, but may be a protected hydroxyl group in the starting material and modified during the process,

R ¹² in the final product may be the same as in the starting material or may be altered during the process,

R ¹² in the final product is as defined above, and if it is a hydroxyl group, is obtained from a cyano group during the process,

R ^a is as defined above and can then be acylated as a step of the process to obtain the end product with an acylated R ^a group.

Preferably, in the starting material, R ⁵ is acetyl or another small acyl group and does not change during the course of the reaction. Preferably, the 15 in the starting material R ¹⁸ is a hydroxyl group and does not change during the reaction. It is preferred that the starting material R ¹² is CH ₃ and does not change during the reaction. In the final product, R ²¹ is as defined above and, if a hydroxyl group, is obtained from a cyano group during the process. Preferably, in the end product R ^a is as defined for the compound of formula (XXIII).

Another important method of the present invention involves the interaction of:

Another important method of the present invention involves the interaction of:

Another important method of the present invention involves an interaction wherein the R ¹ group is aminomethylene and converted to a hydroxymethylene group.

Another important method of the present invention involves reacting wherein the compound of group R ¹ which is hydroxymethylene is reacted with a reagent of formula (XIX):

65893 Bl in which Fu is a protected functional group, Prot ³ is a protecting group and the dotted line indicates a possible double bond.

Another important method of the present invention involves reacting to obtain a 21-cyan compound of formula (XVI), characterized in that it involves reacting a compound of formula (XV):

OCH ₃ R ^1s xA / CH ₃

I ⁵ n A ^£ 1 ^CH h \ /xJ.XLY^R ¹⁵

T and II 7 J ^CH 3

AA Ji M ” ^{,, R14a} ch ₃ o γγ 'r R ^14b

R8 is R21 in which the groups R ¹ , R ⁵ , R ⁸ , R ^14a , R ^14b , R ¹⁵ and R ¹⁸ are as defined above and R ²¹ is a hydroxyl group having a cyanide ion source to form the desired 21-cyan compound.

In addition, methods using other nucleophil-containing compounds to obtain similar compounds of formula (XVI) in which position 21 is protected by another nucleophilic group, the 21-NuC group, are also contemplated. For example, a 21-Nuc compound of formula (XVI) with an alkylamino substituent at the 21-position can be prepared by reacting a compound of formula (XV) in which R ²¹ is a hydroxyl group, with a suitable alkylamine. The 21-Nuc compound of formula (XVI) with an alkylthio substituent at the 21-position can also be prepared by reacting the compound of formula (XV), in which R ²¹ is a hydroxyl group, with a suitable alkanethiol. Alternatively, the 21 -Nuc compound of formula (XVI) with an alpha-carbonylalkyl substituent at the 21-position can be prepared by reacting the compound of formula (XV) in which R ²¹ is a hydroxyl group, with a suitable carbonyl compound, usually in the presence of a base. Other synthetic methods are known for other 21-Nuc compounds.

Another important reaction of the present invention involves treating the 21-cyan compound of the present invention to give the 21-hydroxyl compound. These compounds have interesting in vivo properties.

Examples of carrying out the invention

Example 1

To a solution of 2 (21.53 g, 39.17 mmol) in ethanol (200 ml) was added tert-butoxycarbonyl anhydride (7.7 g, 35.25 mmol) and the mixture was stirred for 7 h at 23 ° C. The reaction mixture was then concentrated in vacuo and the residue purified by flash chromatography (silica gel, hexane: ethyl acetate 6: 4) to give 14 (20.6 g, 81%) as a yellow solid.

Rf: 0.52 (ethyl acetate: chloroform 5: 2).

Ή NMR (300 MHz, CDC1 _3): delta 6.49 (s, IH), 6.32 (bs, IH), 5.26 (bs, IH), 4.60 (bs, IH), 4.14 (d,> 2.4 Hz, IH), 4.05 (d, J = 2.4 Hz, 1H), 3.94 (s, 3H), 3.81 (d,> 4.8 Hz, 1H), 3.7 (s, 3H), 3.34 (brd,> 7.2Hz, 1H), 3.18- 3.00 (m, 5H), 2.44 (d,> 18.3 Hz, 1H), 2.29 (s, 3H), 2.24 (s, 3H), 1.82 (s, 3H), 1.80-1.65 (m, 1H), 1.48 ( s, 9H), 0.86 (d,> 5.7Hz, 3H).

65893 ^B13 C NMR (75 MHz, CDCl3): delta 185.5,

180.8, 172.7, 155.9, 154.5, 147.3, 143.3, 141.5,

135.3, 130.4, 129.2, 127.5, 120.2, 117.4, 116.9,

80.2, 60.7, 60.3,58.5, 55.9,55.8, 54.9,54.4,50.0,

41.6, 40.3, 28.0, 25.3, 24.0, 18.1, 15.5, 8.5. ESI-MS m / z: Calculated for C ₃₄ H ₄₃ O ₈ : 649.7.

Found (M + H) ⁺ : 650.3.

Example 2

With stirring to a solution of 14 (20.6 g, 31.75 mmol) in CH ₃ CN (159 ml), diisopropylethylamine (82.96 ml, 476.2 mmol), methoxymethylene bromide (25.9 ml, 317.5 mmol) and dimethylaminopyridine (155 mg) were added at 0 ° C. , 1.27 mmol). The mixture was stirred at 23 ° C for 24 h. The reaction was fixed at 0 ° C with aqueous 0.1 N hydrochloric acid (750 ml) (pH = 5) and extracted with methylene chloride (2x400 ml). The organic phase was dried over sodium sulfate and concentrated in vacuo. The residue was purified by flash chromatography (silica gel, gradient from hexane: ethyl acetate 4: 1 to hexane: ethyl acetate 3: 2) to give 15 (17.6 g, 83%) as a yellow solid.

Rf: 0.38 (hexane-etidacetate 3: 7). ³ H NMR (300 MHz, CDCl ₃ ): delta 6.73 (s, 1H), 5.35 (bs, 1H), 5.13 (s, 2H), 4.40 (bs, 1H), 4.25 (d, J = 2.7 Hz, 1H) , 4.03 (d, J = 2.7 Hz, 1H), 3.97 (s, 3H), 3.84 (bs, 1H), 3.82-3.65 (m, 1H), 3.69 (s, 3H), 3.56 (s, 3H). 3.39-3.37 (m, 1H), 3.20-3.00 (m, 5H), 2.46 (d, J = 18 Hz, 1H), 2.33 (s, 3H), 2.23 (s, 3H), 1.85 (s, 3H) , 1.73-1.63 (m, 1H), 1.29 (s, 9H), 0.93 (d, J = 5.1 Hz, 3H).

¹³ C NMR (75 MHz, CDCl ₃ ): delta 185.4,

180.9, 172.4, 155.9, 154.5, 149.0, 148.4, 141.6, 135.1,131.0,129.9,127.6,124.4,123.7,117.3,99.1,

79.3, 60.7, 59.7, 58.4, 57.5, 56.2, 55.9, 55.0, 54.2, 50.0, 41.5, 39.9,28.0, 25.2, 24.0,18.1, 15.6, 8.5.

ESI-MS m / z: Calculated for C..HO: 693.8.

Z 47 9

Found (M + H) ⁺ : 694.3.

Example 3

65893 Bl

To a flask containing 15 (8 g, 11.5 mmol) in methanol (1.61), at 0 ° C, was added 1M aqueous sodium hydroxide solution (3.2 1). The reaction mixture was stirred for 2 h at this temperature and then fixed with 6M hydrochloric acid to pH 5. The mixture was extracted with ethyl acetate (3x11) and the combined organic layers dried over sodium sulfate and concentrated in vacuo. The residue was purified by flash chromatography (silica gel, gradient chloroform: ethyl acetate 2: 1) to give 16 (5.3 g, 68%).

Rf: 0.48 (CH ₃ SC: water 7: 3, RP-C18). Ή NMR (300 MHz, CDC1 _3): delta 6.73 (s, IH), 5.43 (bs, IH), 5.16 (s, 2H), 4.54 (bs, IH), 4.26 (d, J = 1.8 Hz, IH) , 4.04 (d,> 2.7 Hz, 1H), 3.84 (bs, 1H), 3.80-3.64 (m, 1H), 3.58 (s, 3H), 3.41-3.39 (m, 1H), 3.22-3.06 (m. 5H), 2.49 (d,> 18.6 Hz, 1H),

2.35 (s, 3H), 2.30-2.25 (m, 1H), 2.24 (s, 3H), 1.87 (s, 3H), 1.45-1.33 (m, 1H), 1.19 (s, 9H), 1.00 (br d ,> 6.6 Hz, 3H).

¹³ C NMR (75 MHz, CDCl ₃ ): delta 184.9,

180.9, 172.6, 154.7, 151.3 149.1, 148.6, 144.7,

132.9, 131.3, 129.8, 124.5, 123.7, 117.3, 116.8,

99.1, 79.4, 59.8, 58.6,57.7, 56.2, 55.6, 54.9,54.5,

50.1, 41.6, 40.1, 28.0, 25.3, 24.4, 18.1, 15.7, 8.0.

ESI-MS m / z: Calculated for C ₃₅ H _{4 J} N ₅ O ₉ :

679.7. Found (M + H) ⁺ : 680.3.

Example 4

17

To a degassed solution of compound 16 (1.8 g, 2.64 mmol) in dimethylformamide (221 ml) was added 10% palladium on carbon (360 mg) and stirred under hydrogen (atmospheric pressure) for 45 min. The reaction mixture was filtered through Celite iodine argon in a flask containing anhydrous cesium carbonate (2.58 g, 7.92 mmol). Bromochloromethane (3.40 ml, 52.8 mmol) was then added and the vessel was dissolved and stirred at 100 ° C for 2 h. The reaction mixture was cooled, filtered through a pad of celite and washed with methylene chloride. The organic layer was concentrated and dried over sodium sulfate to give 17 as a brown oil, which was used in the next step without further purification.

Rf: 0.36 (hexane: ethyl acetate 1: 5, silica gel). NMR (300 MHz, CDCl ₃ ): delta 6.68 (s, 1H), 6.05 (bs, 1H), 5.90 (s, 2H), 5.79 (s, 1H), 5.40 (bs, 1H), 5.31-5.24 ( m, 2H), 4.67 (d,> 8.1 Hz, 1H), 4.19 (d,> 2.7 Hz, 1H), 4.07 (bs, 1H), 4.01 (bs, 1H), 3.70 (s, 3H), 3.67 ( s, 3H), 3.64-2.96 (m, 5H), 2.65 (d,> 18.3 Hz, 1H), 2.33 (s, 3H), 2.21 (s, 3H), 2.04 (s, 3H), 2.02-1.95 ( m, 1H), 1.28 (s, 9H), 0.87 (d,> 6.3 Hz, 3H).

¹³ C NMR (75 MHz, CDCl ₃ ): delta 172.1,

162.6, 154.9, 149.1, 145.7, 135.9, 130.8, 130.7,

125.1, 123.1, 117.8, 100.8, 99.8, 76.6, 59.8, 59.2, 57.7,57.0,56.7,55.8,55.2,49.5,41.6,40.1,36.5, 31.9,31.6,29.7,28.2,26.3,25.0,22.6,18.2, 15.8,

14.1, 8.8.

ESI-MS m / z: Calcd. For C ₃₆ H ₄₇ N ₅ O ₉ : 693.34. Found (M + H) ⁺ : 694.3.

65893 Bl

Example 5

To a flask containing a solution of 17 (1.83 g, 2.65 mmol) in dimethylformamide (13 ml), at 0 ° C was added cesium carbonate (2.6 g, 7.97 mmol) and allyl bromide (1.15 ml, 13.28 mmol). The resulting reaction mixture was filtered through a pad of celite and washed with methylene chloride. The organic layer was dried over sodium sulfate and concentrated. The residue was purified by flash chromatography (silica gel, chloroform: ethyl acetate 1: 4) to give 18 (1.08 g, 56%) as a white solid.

Rf: 0.36 (chloroform: ethyl acetate 1: 3). NMR (300 MHz, CDCl3): delta 6.70 (s, 1H), 6.276.02 (m, 1H), 5.94 (s, 1H), 5.83 (s, 1H), 5.37 (dd, J = 1.01 Hz, J = 16.8 Hz, 1H), 5.40 (bs, 1H), 5.25 (dd, J = 1.0Hz, J = 10.5Hz, 1H), 5.10 (s, 2H), 4.91 (bs, 1H), 4.25-4.22 (m , 1H), 4.21 (d, J = 2.4 Hz,

1H), 4.14-4.10 (m, 1H), 4.08 (d, J = 2.4 Hz, 1H), 4.00 (bs, 1H), 3.70 (s, 3H), 3.59 (s, 3H), 3.563.35 (m , 2H), 3.26-3.20 (m, 2H), 3.05-2.96 (dd, ²⁰ J = 8.1 Hz, J = 18 Hz, 1H), 2.63 (d, J = 18 Hz, 1H).

2.30 (s, 3H), 2.21 (s, 3H), 2.09 (s, 3H), 1.91-1.80 (m, 1H), 1.24 (s, 9H), 0.94 (d, J = 6.6 Hz, 3H).

¹³ C NMR (75 MHz, CDCl ₃ ): delta 172.0,

154.8, 148.8, 148.6, 148.4, 144.4, 138.8, 133.7, ²⁵ 130.9, 130.3, 125.1, 124.0, 120.9, 117.8, 112.8,

112.6, 101.1, 99.2, 73.9, 59.7, 59.3, 57.7, 56.9,

56.8,56.2,55.2,40.1,34.6,31.5,28.1,26.4,25.1,

22.6, 18.5, 15.7, 14.0, 9.2.

ESI-MS m / z: Calculated for C, _o H ₍₁ N, 0 ": ol 39 51 5 9

733.4. Found (M + H) ⁺ : 734.4.

Example 6

4.3M HCl / dioxane

12h ', 23 ° C

65893 Bl

To a solution of 18 (0.1 g, 0.137 mmol) in dioxane (2 ml) was added 4.2 M hydrochloric acid / dioxane (1.46 ml) and the mixture was stirred for 1.2 h at 23 ° C. The reaction mixture was fixed at 0 ° C with saturated aqueous sodium bicarbonate solution (60 ml) and extracted with ethyl acetate (2x70 ml). The organic layer was dried over sodium sulfate and concentrated in vacuo to give 19 (267 mg, 95%) as a white solid, which was used in the next step without further purification.

Rf: 0.17 (ethyl acetate: methanol 10: 1, silica gel).

Ή NMR (300 MHz, CDC1 _3): delta 6.49 (s,

1H), 6.12-6.00 (m, 1H), 5.94 (s, 1H), 5.86 (s, 1H),

5.34 (dd, J, = 1.0 Hz, J ₂ = 17.4 Hz, 1H), 5.25 (dd, Jj = 1.0 Hz, J = 10.2 Hz, 1H), 4.18-3.76 (m, 5H).

3.74 (s, 3H), 3.71-3.59 (m, 1H), 3.36-3.20 (m, 5 4H), 3.01-2.90 (m, 1H), 2.60 (d, J = 18.0 Hz, 1H).

2.29 (s, 3H), 2.24 (s, 3H), 2.11 (s, 3H), 1.97-1.86 (m, 1H), 0.93 (d, J = 8.7 Hz, 3H).

¹³ C NMR (75 MHz, CDCl ₃ ): delta 175.5,

148.4, 146.7, 144.4, 142.4, 138.9, 133.7, 131.3, 10 128.3, 120.8, 117.9, 117.4, 113.8, 112.4, 101.1,

74.2,60.5,59.1,56.5,56.1,56.3,56.0,55.0,50.5,

41.6, 39.5, 29.5, 26.4, 24.9, 21.1, 15.7, 9.33.

ESI-MS m / z: Calculated for C ₃₂ H ₃₉ N ₅ O _6: 589.

Found (M + H) ⁺ : 590.

Example 7

NH ₂ phenylisothiocyanate

1.23 * 0

Me

Me

To a solution of 19 (250 mg, 0.42 mmol) in methylene chloride (1.5 ml) was added phenylisothiocyanate (0.3 ml, 2.51 mmol) and the mixture was stirred at 23 ° C for 1 h. The reaction mixture was concentrated in vacuo and the residue purified by flash chromatography (silica gel, hexane to 5: 1 hexane: ethyl acetate) to give 20 (270 mg, 87%) as a white solid.

Rf: 0.56 (chloroform: ethyl acetate 1: 4). Ή NMR (300 MHz, CDCl 3: δ 8.00 (bs, 1H), 7.456.97 (m, 4H), 6.10 (s, 1H), 6.08-6.00 (m, 1H), 5.92 (s, 1H), 5.89 ( s, 1H), 5.82 (s, 1H), 5.40 (dd, J = 1.5 Hz, J = 17.1 Hz, 1H), 5.38 (bs, 1H), 5.23 (dd, J = 1.5 Hz, J = 10.5 Hz. 1H), 4.42-4.36 (m, 1H), 4.19-4.03 (m, 5H), 3.71 (s, 3H), 3.68-3.17 (m, 4H), 2.90 (dd, J = 7.8 Hz, J = 18.3 Hz , 1H), ³⁰ 2.57 (d, J = 18.3 Hz, 1H), 2.25 (s, 3H), 2.12 (s,

3H), 2.10 (s, 3H), 1.90 (dd, J = 12.3 Hz, J = 16.5 Hz, 1H), (m, 1H), 0.81 (d, J = 6.9 Hz, 3H).

¹³ C NMR (75 MHz, CDCl ₃ ): delta 178.4,

171.6, 148.6, 146.8, 144.3, 142.7, 138.7, 136.2, 35 133.6, 130.7, 129.8, 126.6, 124.2, 124.1, 120.9,

117.7, 117.4., 116.7, 112.6, 112-5, 101.0, 74.0,

60.6, 59.0,57.0,56.2, 56.1,55.0,53.3,41.4,39.7,

26.3, 24.8, 18.3, 15.5, 9.2.

ESI-MS m / z: Calculated for C ₃₀ H 2 NO:

44 6 6 ₄₀ 724.8. Found (M + H) ⁺ : 725.3.

Example 8

65893 Bl

To a solution of 20 (270 mg, 0.37 mmol) in dioxane (1 ml) was added 4.2M hydrochloric acid / dioxane (3.5 ml) and the mixture was stirred at 23 ° C for 30 min. Then ethyl acetate (20 ml) and water (20 ml) were added and the organic layer was decanted. The aqueous phase was basified at 0 ° C with saturated aqueous sodium bicarbonate solution (60 ml) (pH 8) and then extracted with methylene chloride (2x50 ml). The combined organic extracts were dried over sodium sulfate and concentrated in vacuo. The residue was purified by flash chromatography (silica gel, ethyl acetate: methanol 5: 1) to give 21 (158 mg, 82%) as a white solid.

Rf: 0.3 (ethyl acetate: methanol 1: 1). 1 H NMR (300 MHz, CDCl ₃ ): delta 6.45 (s, 1H), 6.12-6.03 (m, 1H), 5.91 (s, 1H), 5.85 (s, 1H), 5.38 (dd, J = 1.2)

Hz, J = 17.1 Hz, 1H), 5.24 (dd, J = 1.2Hz, J = 10.5

Hz, 1H), 4.23-4.09 (m, 4H), 3.98 (d, J = 2.1 Hz, 5H), 3.90 (bs, 1H), 3.72 (s, 3H), 3.36-3.02 (m.

5H), 2.72-2.71 (m, 2H), 2.48 (d, J = 18.0 Hz, 1H),

2.33 (s, 3H), 2.22 (s, 3H), 2.11 (s, 3H), 1.85 (dd, J = 11.7 Hz, J = 15.6Hz, 1H).

¹³ C NMR (75 MHz, CDCI,): delta 148.4,

146.7, 144.4, 142.8, 138.8, 133.8, 130.5, 128.8,

121.5, 120.8, 118.0, 117.5, 116.9, 113.6, 112.2,

101.1, 74.3, 60.7, 59.9, 58.8, 56.6, 56.5, 55.3,

44.2, 41.8, 29.7, 26.5, 257, 15.7, 9.4.

ESI-MS m / z: Calculated for C ₂₉ H ₃₄ N ₄ O ₅ : 15 518.3. Found (M + H) ⁺ : 519.2.

Example 9

To a solution of 21 (0.64 g, 1.22 mmol) in methylene chloride (6.13 ml) at -10 ° C was added pyridine (0.104 ml, 1.28 mmol). The mixture was stirred at this temperature for 1 h, then fixed by the addition of 0.1 N hydrochloric acid (10 ml) and extracted with methylene chloride (2 x 10 ml). The organic layer was dried over sodium sulfate and concentrated in vacuo. The residue was purified by flash chromatography (silica gel, hexane: ethyl acetate 1: 2) to give 22 (0.84 g, 98%) as a white porous solid.

Rf: 0.57 (ethyl acetate: methanol 5: 1). 1 H NMR (300 MHz, CDCl ₃ ): delta 6.50 (s, 1H), 6.10-6.00 (m, 1H), 6.94 (d, J = 1.5 Hz, 1H), 5.87 (d, J = 1.5)

Hz, 1H), 5.73 (bs, 1H), 5.37 (dq, J = 1.5Hz, J = 17.1

Hz, 1H), 5.26 (dq, J = 1.8 Hz, J = 10.2 Hz, 1H), 4.60 (d, J = 12 Hz, 1H), 4.22-4.10 (m, 4H), 4.19 (d, J = 12 Hz, 1H), 4.02 (m, 2H), 3.75 (s, 3H), 3.373.18 (m, 5H), 3.04 (dd, J = 8.1 Hz, J = 18 Hz, 1H), 30 2.63 (d. J = 18 Hz, 1H), 2.31 (s, 3H), 2.26 (s, 3H),

2.11 (s, 3H), 1.85 (dd, J = 12.3 Hz, J = 15.9 Hz, 1H).

¹³ C NMR (75 MHz, CDCl 3): delta 154.3,

148.5, 146.7, 144.5, 142.8, 139.0, 138.8, 130.7,

128.7, 121.3, 120.8, 117.8, 117.7, 116.8, 112.7, 35 101.2, 77.2, 74.3, 60.7, 59.9, 57.0, 56.4, 55.3,

43.3, 41.7, 31.6, 26.4, 25.3, 22.6, 15.9, 14.1, 9.4.

ESI-MS m / z: Calculated for C ₃₂ H ₃₅ C1 ₃ N ₄ O ₇ :

694.17. Found (M + H) ⁺ : 695.2.

Example 10

65893 Bl

To a solution of 22 (0.32 g, 0.46 mmol) in CH ₃ CN (2.33 ml) at 0 ° C was added diisopropylethylamine (1.62 ml, 9.34 mmol), bromomethylmethyl ether (0.57 ml, 7.0 mmol) and dimethylaminopyridine (6 mg, 0.046 mmol). The mixture was stirred at 30 ° C for 10 h. The reaction mixture was then diluted with dichloromethane (30 ml) and poured into aqueous hydrochloric acid (10 ml) (pH = 5). The organic layer was dried over sodium sulfate and the solvent was removed under reduced pressure to give a residue which was purified by flash chromatography (silica gel, hexane: ethyl acetate 2: 1) to give 23 (0.304 g, 88%) as white porous solid.

Rf: 0.62 (hexane: ethyl acetate 1: 3). 1 H NMR (300 MHz, CDCl ₃ ): delta 6.73 (s, 1H), 6.10 (m, 1H), 5.94 (d, J = 1.5 Hz, 1H), 5.88 (d,> 1.5 Hz, 1H), 5.39 ( dq, J = 1.5 Hz, J = 17.1 Hz, 1H), 5.26 (dq, J = 1.8Hz, J = 10.2Hz, 1H), 5.12 (s, 2H), 4.61 (d,> 12 Hz, 1H). 4.55 (t,> 6.6 Hz, 1H), 4.25 (d,> 12 Hz, 1H), 4.22-4.11 (m, 4H), 4.03 (m, 2H), 3.72 (s, 3H), 3.58 (s, 3H) ), 3.38-3.21 (m, 5H), 3.05 (dd, J = 8.1 Hz, J = 18 Hz, 1H), 2.65 (d,> 18 Hz, 1H), 2.32 (s, 3H), 2.23 (s. 3H), 2.12 (s, 3H), 1.79 (dd, J = 12.3 Hz, J = 15.9 Hz, 1H).

¹³ C NMR (75 MHz, CDCl ₃ ): delta 154.3,

148.6, 148.4, 144.5, 139.0, 133.6, 130.6, 130.1, 125.07, 124.7, 124.0, 121.1, 117.7, 112.6, 101.2,

99.2,77.2,74.4,74.1,59.8,59.8,57.7, 57.0,56.8, 56.68, 55.3, 43.2, 41.5, 26.4, 25.2, 15.9, 9.3.

ESI-MS m / z: Calculated for C ₃₄ H ₃₉ Cl ₃ N ₄ O ₈ : 738.20. Found (M + H) ⁺ : 739.0.

Example 11

AsON aq. Zn

Yh, 23 ° C

To a suspension of 23 (0.304 g, 0.41 mmol) in 90% aqueous acetic acid (4 ml) was added powdered zinc (0.2 g, 6.67 mmol) and the reaction mixture was stirred at 23 ° C for 7 h. The mixture was filtered through a pad of celite, which was washed with dichloromethane. The organic layer was washed with aqueous saturated sodium bicarbonate solution (15 ml) (pH = 9) and dried over sodium sulfate. The solvent was removed under reduced pressure to give 24 (0.191 g, 83%) as a white solid.

Rf: 0.3 (ethyl acetate: methanol 5: 1). 1 H NMR (300 MHz, CDCl ₃ ): delta 6.68 (s, 1H), 6.09 (m, 1H), 5.90 (d,> 1.5 Hz, 1H), 5.83 (d,> 1.5 Hz 1H), 5.39 (dq. J = 1.5 Hz, J = 17.1 Hz, 1H), 5.25 (dq, J = 1.5 Hz, J = 10.2 Hz, 1H), 5.10 (s, 2H), 4.224.09 (m, 3H), 3.98 (d. J = 2.4 Hz, 1H), 3.89 (m,

1H), 3.69 (s, 3H), 3.57 (s, 3H), 3.37-3.17 (m, 3H),

3.07 (dd, J = 8.1 Hz, J = 18 Hz, 1H), 2.71 (m, 2H),

2.48 (d,> 18 Hz, 1H), 2.33 (s, 3H), 2.19 (s, 3H),

2.17 (s, 3H), 1.80 (dd, J = 12.3 Hz, J = 15.9 Hz, 1H).

¹³ C NMR (75 MHz, CDCl ₃ ): delta 148.5,

148.2, 144.3, 138.7, 133.7, 130.7, 129.9, 125.0,

123.9, 121.3, 117.9, 117.5, 113.6, 112.0, 101.0,

99.2, 74.0, 59.8, 59.7,58.8, 57.6, 57.0, 56.2, 55.2,

44.2, 41.5, 31.5, 26.4, 25.6, 22.5, 16.7, 14.0, 9.2.

ESI-MS m / z: Calculated for C ₃₁ H _3g N ₄ O ₆ :

562.66. Found (M + H) ⁺ : 563.1.

Example 12

65893 Bl

To a solution of 24 (20 mg, 0.035 mmol) in water (0.7 ml) and tetrahydrofuran (0.7 ml) at 0 ° C was added sodium nitrite (1.62 mg, 0.17 mmol) and 90% aqueous acetic acid (0.06 ml) and the mixture. was stirred at 0 ° C for 3 h. The reaction mixture was then diluted with dichloromethane (5 ml) and the organic layer was washed with water (1 ml), dried over sodium sulfate and concentrated in vacuo. The residue was purified by flash chromatography (silica gel, hexane: ethyl acetate 2: 1) to give 25 (9.8 mg, 50%) as a white solid.

Rf: 0.34 (hexane: ethyl acetate 1: 1). 1 H NMR (300 MHz, CDCl 3): delta 6.71 (s, 1H), 6.11 (m, 1H), 5.92 (d,> 1.5 Hz, 1H), 5.87 (d,> 1.5 Hz,

1H), 5.42 (dq, J = 1.5 Hz, J = 17.1 Hz, 1H), 5.28 (dq, J = 1.8Hz, J = 10.2Hz, 1H), 5.12 (s, 2H), 4.264.09 (m. 3H), 4.05 (d,> 2.4 Hz, 1H), 3.97 (t,> 3.0 Hz, 1H), 3.70 (s, 3H), 3.67-3.32 (m, 4H), 3.58 (s, 5 3H), 3.24 (dd, J = 2.7 Hz, J = 15.9 Hz, 1H), 3.12 (dd, J = 8.1 Hz, J = 18.0Hz, 1H), 2.51 (d,> 18Hz,

1H), 2.36 (s, 3H), 2.21 (s, 3H), 2.12 (s, 3H), 1.83 (dd, J = 12.3 Hz, J = 15.9 Hz, 1H).

¹³ C NMR (75 MHz, CDCl 3): delta 148.7, 10 148.4, 138.9, 133.7, 131.1, 129.4, 125.1, 123.9,

120.7, 117.6, 117.5, 113.2, 112.3, 101.1, 99.2,

74.0,63.2,59.8,59.7,57.9,57.7,57.0,56.5,55.2,

41.6, 29.6, 26.1, 25.6, 22.6, 15.7, 9.2.

ESI-MS m / z: Calculated for C ₃₁ H ₃₇ N ₃ O ₇ : 15 563.64. Found (M + H) ⁺ : 564.1.

Example 13

years ίι Mr- Oh Yr 2,2,2-Trichloroethyi chloroformate , Α NH3C1 NaH, THF, reflux NHTroc / == \ 4) XJ

The starting material (2.0 g, 5.90 mmol) was added to a suspension of sodium hydride (354 mg, 8.86 mmol) in tetrahydrofuran (40 ml) at 23 ° C, after which the suspension was treated with allyl chloroformate (1.135 ml, 8.25 mmol) at 23 And then heated at reflux for 3 h. The suspension was cooled, filtered, the solid was washed with ethyl acetate (100 ml) and the filtrate was concentrated. The crude oil was stirred with hexane (100 ml) and kept at 4 ° C overnight. The solvent was then decanted and the light yellow slurry treated with dichloromethane (20 mi) and precipitated with hexane (100 ml). After 10 min, the solvent was decanted again. The operation is repeated until a white solid appears. The white solid was filtered off and dried to give 29 (1.8 g, 65%) as a white solid.

NMR (300 MHz, CDCl ₃ ): delta 7.74 (d,> 7.5 Hz, 2H), 7.62 (d,> 6.9 Hz, 2H), 7.33 (t,> 7.5 Hz, 2H), 7.30 (t,> 6.3 Hz, 2H), 5.71 (d,> 7.8 Hz, 1H), 4.73 (d,> 7.8 Hz, 2H), 4.59 (m, 1H), 4.11 (t,> 6.0 Hz, 1H), 3.17 (dd, J = 6.0 Hz, J = 2.7 Hz, 2H), 3.20 (dd, J = 5.4 Hz, J = 2.1 Hz, 2H).

¹³ C NMR (75 MHz, CDCl ₃ ): delta 173.6,

152.7, 144.0, 139.7, 137.8, 126.0, 125.6, 123.4,

118.3, 73.4, 52.4, 45.5, 35.8, 33.7.

ESI-MS m / z: Calculated for C ₂₀ H ₁₈ Cl ₃ NO ₄ S:

474.8. Found (M + Na) ⁺ : 497.8.

Example 14

Me

A mixture of compound 25 (585 mg, 1.03 mmol) and compound 29 (1.47 g, 3.11 mmol) was azeotroped with anhydrous toluene (3x10 ml). To a solution of 25 and 29 in anhydrous dichloromethane (40 ml) was added DMAP (633 mg, 5.18 mmol) and EDC hydrochloride (994 mg, 5.18 mmol) at 23 ° C. The reaction mixture was stirred at 23 ° C for 3 h. The mixture was separated using a saturated aqueous sodium bicarbonate solution

65893 B1 (50 ml) and the layers were separated. The aqueous layer was washed with dichloromethane (50 ml). The combined organic layers were dried over sodium sulfate, filtered and concentrated. The crude product was purified by flash chromatography (silica gel, hexane: ethyl acetate 3: 1) to give 30 (1.00 g, 95%) as a pale yellow creamy solid.

Ή NMR (300 MHz, CDCl ₃ ): δ 7.72 (m, 2H), 7.52 (m, 2H), 7.38 (m, 2H), 7.28 (m, 2H), 6.65 (s, 1H), 6.03 (m, 1H), 5.95 (d,> 1.5 Hz, 1H), 5.79 (d,> 1.5 Hz, 1H), 5.39 (m, 1H), 5.29 (dq, J = 10.3 Hz,> 1.5 Hz, 1H), 5.10 ( s, 2H), 4.73 (d,> 11.9 Hz, 1H), 4.66 (d,> 11.9 Hz, 1H), 4.53 (m, 1H), 4.36-3.96 (m, 9H), 3.89 (t,> 6.4 Hz ,

1H), 3.71 (s, 3H), 3.55 (s, 3H), 3.33 (m, 1H), 3.20 (m, 2H), 2.94 (m, 3H), 2.59 (m, 1H), 2.29 (s, 3H) ), 2.23 (s, 3H), 2.02 (s, 3H), 1.83 (dd, J = 16.0 Hz, J = 11.9 Hz, 1H).

¹³ C NMR (75 MHz, CDCl ₃ ): delta 169.7,

154.0, 148.8, 148.4, 145.7, 144.5, 140.9, 139.0, 133.7, 130.9, 130.6, 127.6, 127.0, 124.8, 124.6,

124.1, 120.8, 119.9, 118.2, 117.7, 117.3, 112.7,

112.1, 101.3, 99.2, 74.7, 73.9, 64.4, 59.8, 57.7,

57.0, 56.8,55.4,53.3,46.7,41.4,36.5,34.7,31.5,

26.4, 24.9, 22.6, 15.7, 14.0, 9.1.

ESI-MS m / z: Calculated for

C ₅₁ H ₅₃ Cl ₃ N ₄ O ₁₀ S: 1020.4. Found (M + H) ⁺ :

1021.2.

Example 15

31

To a solution of 30 (845 mg, 0.82 mmol), acetic acid (500 mg, 8.28 mmol) and (PPh ₃ ) ₂ PdCl ₂ (29 mg, 0.04 mmol) in anhydrous dichloromethane (20 ml) was added at 23 ° C. dropwise Bu ₃ SnH (650 mg, 2.23 mmol). The reaction mixture was stirred at this temperature for 15 min, whereby bubbles were removed. The crude product was fixed with water (50 ml) and extracted with dichloromethane (3x50 ml). The organic layers were dried over sodium sulfate, filtered and concentrated. The crude product was purified by flash chromatography (ethyl acetate / hexane in a gradient of 1: 5 to 1: 3) to give compound 31 (730 mg, 90%) as a pale yellow creamy solid.

Ή NMR (300 MHz, CDCl ₃ ): δ 7.72 (m, 2H), 7.56 (m, 2H), 7.37 (m, 2H), 7.30 (m, 2H),

6.65 (s, 1H), 5.89 (s, 1H), 5.77 (s, 1H), 5.74 (s, 1H), 5.36 (d,> 5.9 Hz, 1H), 5.32 (d,> 5.9 Hz, 1H). 5.20 (d,> 9.0 Hz, 1H), 4.75 (d,> 12.0 Hz, 1H), 4.73 (m, 1H), 4.48 (d,> 11.9 Hz, 1H), 4.08 (m, 4H), 3.89 (m , 1H), 3.86 (t,> 6.2 Hz, 1H), 3.70 (s, 3H), 3.69 (s, 3H), 3.38 (m, 1H), 3.25 (m, 1H), 3.02-2.89 (m, 4H) ), 2.67 (s, 1H), 2.61 (s, 1H), 2.51 (dd, J = 14.3 Hz, J ₂ = 4.5 Hz, 1H), 2.29 (s, 3H), 2.23 (s, 3H), 1.95 ( s, 3H), 1.83 (m, 1H).

¹³ C NMR (75 MHz, CDCl ₃ ): delta 168.2,

152.5, 148.1, 146.2, 144.4, 144.3, 143.3, 134.6,

129.7, 129.6, 126.2, 125.6, 123.4, 123.3, 121.6,

118.5.116.3.110.2, 105.1, 99.4, 98.5, 75.2, 73.3,

61.7, 58.4, 57.9,56.3, 56.1, 55.1,54.7, 53.9, 51.9,

45.2, 40.1, 35.6, 33.3, 24.8, 23.3, 14.5, 7.3.

ESI-MS m / z: Calculated for

C ₄₈ H ₄₉ Cl ₃ N ₄ O ₁₀ S: 980.3. Found (M + H) ⁺ : 981.2.

65893 Bl

Example 16

32

To a solution of 31 (310 mg, 0.32 ml) in anhydrous dichloromethane (15 ml), at -10 ° C was added, via cannula, a solution of benzene selenic anhydride 70% (165 mg, 0.32 ml) in anhydrous dichloromethane (7 ml), such as the temperature is maintained at -10 ° C. The reaction mixture was stirred at -10 ° C for 5 min. At this temperature, saturated sodium bicarbonate solution (30 ml) was added. The aqueous solution was washed with more dichloromethane (40 ml). The organic layers were dried over sodium sulfate, filtered and concentrated. The crude product was purified by flash chromatography (ethyl acetate / hexane in a gradient of 1: 5 to 1: 1) to afford compound 32 (287 mg, 91%, HPLC: 91.3%) as a pale creamy solid and as a mixture of two isomers (65:35) to be used in the next step.

Ή NMR (300 MHz, CDC1 _3): delta (mixture of isomers) 7.76 (m, 4H), 7.65 (m, 4H), 7.39 (m, 4H), 7.29 (m, 4H), 6.62 (s, 1H) , 6.55 (s, 1H), 5.7915 5.63 (t, 6H), 5.09 (s, 1H), 5.02 (d,> 6.0 Hz, 1H), 4.99 (d,> 6.0 Hz, 1H), 4.80-4.63 (m , 6H), 4.60 (m, 1H), 4.50 (m, 1H), 4.38 (dd,> 12.8 Hz,> 7.5 Hz, 1H), 4.27 (dd,> 12.8 Hz,> 7.5 Hz, 1H), 4.16- 3.90 (m, 10H), 3.84 (s, 3H), 3.62 (s, 3H),

3.50 (s, 3H), 3.49 (s, 3H), 3.33-2.83 (m, 14H), 2.45-2.18 (m, 2H), 2.21 (s, 6H), 2.17 (s, 6H), 1.77 (s. 6H), 1.67 (m, 2H).

¹³ C NMR (75 MHz, CDCl ₃ ): delta (mixture of isomers) 168.6, 168.4,158.6,154.8,152.8,152.5, 25 147.3, 147.2, 146.8, 144.1, 144.0, 140.8, 139.7,

137.1, 129.8, 129.3, 128.7, 126.5, 125.5, 123.7,

123.6, 123.5, 123.4, 122.2, 121.3, 118.3, 115.8,

110.2.106.9.103.5.103.2.100.1.99.6.97.9.97.7,

93.8,73.4,70.9,69.2,64.9,62.5,59.3,58.9,58.4,

56.7, 56.3, 56.2,55.4, 55.2, 55.1,54.9, 54.7, 54.3,

54.1, 53.8, 52.8,45.5,40.5,40.0,39.8, 35.8, 35.5,

33.9, 33.7,30.1,28.4,24.2,24.1,21.2,14.5,14.4,

12.7, 6.0, 5.7.

ESI-MS m / z: Calculated for

C ₄₈ H ₄₉ Cl ₃ N ₄ O _n S: 996.3. Found (M + H) ⁺ : 997.2.

Example 17

65893 Bl

The reaction vessel was heated twice, purged with argon in vacuo and kept in an argon atmosphere for the reaction. To a solution of dimethyl sulfoxide (39.1 microl, 0.55 mmol, 5 eq) in anhydrous dichloromethane (4.5 ml) at -78 ° C was added dropwise triflic anhydride (37.3 microl, 0.22 mmol, 2 eq). The reaction mixture was stirred at -78 ° C for 20 min, then a solution of 32 (110 mg, 0.11 mmol, HPLC: 91.3%) anhydrous dichloromethane (1 ml for the main addition and 0.5 ml for washing). The temperature was maintained at -78 ° C during the addition and the color changed from yellow to brown. The reaction mixture was stirred at -40 ° C for 35 min. During this period the solution changes from yellow to dark green. IPr ₂ NEt (153 microl, 0.88 mmol, 8 eq) was then added dropwise and the reaction mixture was kept at 0 ° C for 45 min, turning the solution brown. Then t-butanol (41.6 ml, 0.44 mmol, 4 eq) and 2-1-butyl-1,1,3,3 tetramethylguanidine (154.6 microl, 0.77 mmol, 7 eq) were added dropwise and the reaction was stirred at 23 ° C for 40 min. Acetic anhydride (104.3 microl, 1.10 mmol, 10 eq) was then added dropwise and the reaction mixture was maintained at 23 ° C for another 1 h. The reaction mixture was then diluted with dichloromethane (20 ml) and washed with aqueous saturated ammonium chloride solution (50 ml), sodium bicarbonate (50 ml) and sodium chloride (50 ml). The combined organic layers were dried over sodium sulfate, filtered and concentrated. The residue was purified by flash chromatography (ethyl acetate / hexane gradient from 1: 3 to 1: 2) to give compound 33 (54 mg, 58%) as a pale yellow solid.

NMR (300 MHz, CDCl ₃ ): delta 6.85 (s, 1H), 6.09 (s, 1H), 5.99 (s, 1H), 5.20 (d,> 5.8 Hz, 1H), 5.14 (d, J = 5.3) Hz, 1H), 5.03 (m, 1H), 4.82 (d,> 12.2 Hz, 1H), 4.63 (d,> 12.0 Hz, 1H), 4.52 (m, 1H), 4.35-4.17 (m, 4H). 3.76 (s, 3H), 3.56 (s, 3H), 3.45 (m, 2H), 2.91 (m, 2H), 2.32 (s, 3H), 2.28 (s, 3H), 2.21 (s, 3H), 2.12 (m, 2H), 2.03 (s, 3H).

¹³ C NMR (75 MHz, CDCl ₃ ): delta 168.5,

167.2, 152.7, 148.1, 147.1, 144.5, 139.6, 139.1, 130.5, 129.0, 123.7, 123.5, 123.3, 118.8, 116.5,

112.1, 100.6, 97.8, 73.3, 60.5, 59.4, 59.2, 58.3,

57.6, 57.4,56.1,53.3,53.1,40.6,40.0, 31.0,22.2,

18.9, 14.4, 8.1.

ESI-MS m / z: Calculated for C ₃₆ H ₃₉ Cl ₃ N ₄ O _n S: 842.1. Found (M + H) ⁺ : 843.1.

Example 18

To a solution of 33 (12 mg, 0.014 mmol) in anhydrous dichloromethane (1.2 ml) and in pure high-performance liquid chromatography acetonitrile (1.2 ml), sodium iodide (21 mg, 0.14 mmol) was added at 23 ° C and freshly distilled. (above calcium hydride at atmospheric pressure) three methylsilyl chloride (15.4 mg, 0.14 mmol). The reaction mixture turned orange. After 15 minutes, the solution was diluted with dichloromethane (10 ml) and washed with fresh aqueous saturated sodium bisulphite solution (3x10 ml). The organic layer was dried over sodium sulfate, filtered and concentrated. Compound 34 (13 mg, quantitative) was obtained as a pale yellow solid, which was used without further purification.

NMR (300 MHz, CDCl ₃ ): delta 6.85 (s, 1H), 6.09 (s, 1H), 5.99 (s, 1H), 5.27 (d,> 5.8 Hz, 1H), 5.14 (d,> 5.3 Hz) , 1H), 5.03 (d,> 11.9 Hz, 1H), 4.82 (d,> 12.2 Hz, 1H), 4.63 (d,> 13.0 Hz, 1H), 4.52 (m, 1H), 4.34 (m, 1H) , 4.27 (bs, 1H),

65893 Bl

4.18 (m, 2H), 3.76 (s, 3H), 3.56 (s, 3H), 3.44 (m,

1H), 3.42 (m, 1H), 2.91 (m, 2H), 2.32 (s, 3H),

ESI-MS m / z: Calculated for C ₃₄ H ₃₅ N ₄ O ₁₀ S:

798.1. Found (M + H) ⁺ : 799.1.

2.28 (s, 3H), 2.21 (s, 3H), 2.03 (s, 3H).

Example 19

To a solution of 34 (13 mg, 0.016 mmol) in a mixture of acetic acid and water (90: 10.1 ml) was added zinc powder (5.3 mg, 0.081 mniol) at 23 ° C. The reaction mixture was heated at 70 ° C for 6 h. It was then cooled to 23 ° C, diluted with dichloromethane (20 ml) and washed with aqueous saturated sodium bicarbonate solution (15 ml) and aqueous triethylamine solution (15 ml). The organic layer was dried over sodium sulfate, filtered and concentrated. The residue was purified by flash chromatography on Silica-NH ₂ (eluent: ethyl acetate / hexane gradient from 0: 100 to 50:50) to give compound 35 (6.8 mg, 77% in two steps) as a pale yellow solid.

Ή NMR (300 MHz, CDC1 _3): delta 6.51 (s, ¹⁵ IH), 6.03 (dd,> 1.3 Hz,> 26.5 Hz, 2H), 5.75 (bs, IH), 5.02 (d,> 11.6 Hz, IH ), 4.52 (m, 1H), 4.25 (m, 2H), 4.18 (d,> 2.5 Hz, 1H), 4.12 (dd,> 1.9 Hz,> 11.5 Hz, 1H), 3.77 (s, 3H), 3.40 (m, 2H), 3.26 (t,> 6.4 Hz, 1H), 2.88 (m, 2H), 2.30-2.10 ²⁰ (m, 2H), 2.30 (s, 3H), 2.28 (s, 3H), 2.18 ( s, 3H), 2.02 (s, 3H).

^I3 C NMR (75 MHz, CDC1 _3): delta 174.1,

168.4, 147.8, 145.4, 142.9, 140.8, 140.1, 131.7,

130.2, 129.1, 128.3, 120.4, 118.3, 117.9, 113.8, ²⁵ 111.7, 101.7, 61.2, 59.8, 59.2, 58.9, 54.4, 53.8,

54.4, 41.3, 41.5, 34.1, 23.6, 20.3, 15.5, 9.4.

ESI-MS m / z: Calculated for C ₃₁ H ₃₄ N ₄ O _g S:

622.7. Found (M + H) ⁺ : 623.2.

Example 20

To a solution of 36 (49 mg, 0.08 mmol) and 2- [3-hydroxy-4-methoxyphenyl] ethylamine (46.2 mg, 0.27 mmol) in ethanol (2.5 ml) was added silica gel (105 mg) at 23 ° C. The reaction mixture was stirred at 23 ° C for 14 h. 45 Diluted with hexane and poured into a chromatography column (ethyl acetate / hexane 1/3 to 1/1) to give Et-770 (55 mg, 90%) as a pale yellow solid.

Ή NMR (300 MHz, CDCl ₃ ): δ 6.60 (s, 50

1H), 6.47 (s, 1H), 6.45 (s, 1H), 6.05 (s, 1H), 5.98 (s, 1H), 5.02 (d,> 11.4 Hz, 1H), 4.57 (bs, 1H), 4.32 (bs, 1H), 4.28 (d,> 5.3 Hz, 1H), 4.18 (d,> 2.5 Hz, 1H), 4.12 (dd,> 2.1 Hz,> 11.5 Hz, 1H), 3.78 (s, 3H). 3.62 (s, 3H), 3.50 (d,> 5.0 Hz, 1H), 3.42 (m, 1H), 3.10 (ddd,> 4.0 Hz,> 10.0 Hz,> 11.0 Hz, 1H), 2.94 (m, 2H) , 2.79 (m, 1H), 2.61 (m, 1H), 2.47 (m, 1H), 2.35 (m, 1H), 2.32 (s, 3H), 2.27 (s, 3H), 2.20 (s, 3H). 2.09 (m, 1H), 2.04 (s, 3H).

65893 Bl

ESI-MS m / z: Calculated for C ₄₀ H ₄₂ N ₄ O ₁₀ S:

770.7. Found (M + H) ⁺ : 771.2.

Example 22

To a solution of 21 (22 mg, 0.042 mmol) in dichloromethane (0.8 ml) was added phthalic anhydride (6.44 mg, 0.042 mmol) and the reaction mixture was stirred for 2 h at 23 ° C. Carbonyldiimidazole (1 mg, 0.006 mmol) was then added and the mixture was stirred at 23 ° C for 7 h. Carbonyldiimidazole (5.86 mg, 0.035 mmol) was then added and the reaction mixture was stirred for an additional 17 h at 23 ° C. The solution was diluted with dichloromethane (15 ml) and washed with 0.1N hydrochloric acid (15 ml). The organic layer was dried over sodium sulfate, filtered and the solvent was removed under reduced pressure. The residue was purified by flash chromatography (silica, hexane: ethyl acetate 2: 1) to give compound 27 (26.4 mg, 96%) as a white solid.

Rf: 0.58 (ethyl acetate). Ή NMR (300 MHz,

CDC1 ( ₃ ): delta 7.73-7.64 (m, 4H), 6.40 (s, 1H), ¹⁵ 6.12-6.01 (m, 1H), 5.63 (s, 1H), 5.58 (d,> 1.5

Hz, 1H), 5.37 (dd, J = 1.8 Hz, J = 17.4 Hz, 1H), 5.23 (dd, J = 1.8Hz, J = 10.5Hz, 1H), 5.12 (d,> 1.5

Hz, 1H), 4.22-4.15 (m, 3H), 4.08 (d, J = 1.8 Hz,

1H), 3.68 (s, 3H), 3.59-3.55 (m, 2H), 3.35 (d,> 8.1 ²⁰ Hz, 1H), 3.27-3.16 (m, 2H), 3.05 (dd, J = 8.1 Hz.

J = 18.3 Hz, 1H), 2.64 (d,> 18.0Hz, 1H), 2.30 (s, 3H), 2.24 (s, 3H), 2.09 (s, 3H), 1.80 (dd, J = 11.4)

Hz, J = 15 Hz, 1H).

¹³ C NMR (75 MHz, CDCl ₃ ): delta 167.7, ²⁵ 148.9, 146.4, 144.2, 142.6, 139.5, 134.0, 133.5, 132.0, 131.0, 128.3, 123.0, 121.3, 120.9, 118.1,

117.5,116.8,113.6,112.4,100.8,74.5,60.6,60.5,

57.7,56.6, 55.6,55.5,42.3,41.7,26.6,25.5,15.9, 9.46.

3 ° ESI-MS m / z: Calculated for C ₃₇ H ₃₅ N ₄ O ₇ :

648.79. Found (M + H) ⁺ : 649.3.

Example 23

To a solution of 27 (26 mg, 0.041 mmol) in dichloromethane (11 ml) at 23 ° C was added acetic acid (11 ml) and (PPh ₃ ) ₂ PdCl ₂ (2.36 mg) and Bu ₃ SnH (28 ml, 0.10 mmol). The reaction mixture was stirred at this temperature for 2 h and poured into a pressure column (silica, hexane to hexane: ethyl acetate 2: 1) to give 28 (24.7 mg, 99%) as a white solid. Rf: 0.33 (hexane: ethyl acetate 2: 1).

1 H NMR (300MHz, CDCl ₃ ): delta 7.75-7.70 (m, 2H), 7.69-7.65 (m, 2H), 6.39 (s, 1H), 5.82

65893 B1 (bs, 1H), 5.50 (d, J = 1.5 Hz, 1H), 5.0 (d,> 1.5 Hz, 1H), 4.45 (bs, 1H), 4.23-4.19 (m, 2H), 4.10-4.09 (m, 1H), 3.73 (s, 3H), 3.60-3.48 (m, 2H), 3.363.33 (m, 1H), 3.26-3.20 (m, 1H), 3.14-3.08 (m, 1H), 3.98 (d,> 14.4 Hz, 1H), 2.61 (d,> 18.3 Hz, 1H), 2.30 (s, 3H), 2.23 (s, 3H), 2.06 (s, 3H), 1.85 (dd, J = 12 Hz) , J = 15.3 Hz, 1H).

¹³ C NMR (75 MHz, CDCl ₃ ): delta 167.8,

146.4, 145.1, 143.9, 142.7, 137.1, 133.5, 131.9,

130.8, 128.4, 122.9, 120.8, 118.0, 116.8, 114.0,

113.4, 106.4, 100.4, 60.6, 60.5, 57.8, 56.6, 55.6,

55.5, 42.6, 41.5, 25.6, 25.5, 15.8, 8.9.

ESI-MS m / z: Calculated for C ₃₄ H ₃₂ N ₄ O ₇ :

608.6. Found (M + H) ⁺ : 609.2.

Example 24

CH ₃ SOC1, ru. CH ₂ C1 ₂ _

phthalicidin

Acetyl chloride (41.58 microl, 0.58 mmol) and pyridine (47.3 microl, 0.58 mmol) were added to a solution of 28 (357 mg, 0.58 mmol) in dichloromethane (3 ml) at 0 ° C. The reaction mixture was stirred at this temperature for 1 h and then the solution was diluted with dichloromethane (15 ml) and washed with 0.1N hydrochloric acid (15 ml). The organic layer was dried over sodium sulfate, filtered and the solvent was removed under reduced pressure. The residue was purified by flash chromatography (RP-18, CH ₂ CN: H ₂ O 60:40) to give phthalacidin (354 mg, 94%) as white

7.63 (m, 2H), 6.38 (s, 1H), 5.69 (d,> 1.2 Hz, 1H), 5.64 (d,> 1.2 Hz, 1H), 5.30 (bs, 1H), 4.254.21 (m, 2H ), 4.02 (d,> 2.1 Hz, 1H), 3.64-3.62 25 (m, 5H), 3.33 (d,> 8.4 Hz, 1H), 3.21-3.16 (m,

1H), 3.02 (dd, J = 8.1 Hz, J = 18 Hz, 1H), 2.76 (dd,

J = 1.8 Hz, J = 15.6 Hz, 1H), 2.63 (d,> 17.7 Hz, 1H), 2.29 (s, 3H), 2.28 (s, 3H), 2.21 (s, 3H), 2.0 (s, 3H) ), 1.73 (dd, J = 12.0 Hz, J = 15.3 Hz, 1H).

¹³ C NMR (75 MHz, CDCl ₃ ): delta 168.5,

167.6, 146.2, 144.2, 142.5, 141.0, 140.5, 133.4,

131.8, 130.7, 128.2, 120.9, 120.8, 117.9, 116.4,

113.6, 101.1, 60.4, 60.0, 57.0, 56.3, 55.6, 55.4,

41.6, 41.5, 26.5, 25.2, 20.2, 15.7, 9.4.

solid. 35

Rf: 0.37 (CH ₃ CN: H ₂ O 7: 3, RP-18). 1 H NMR (300 MHz, CDCl ₃ ): delta 7.72-7.63 (m, 2H), 7.67ESI-MS m / z: Calcd. For C ₃₆ H ₃₄ N ₄ O ₈ : 650.

Found (M + H) ⁺ : 651.2.

Example 25

65893 Bl

Acetyl chloride (30.7 microl, 0.432 mmol) and pyridine (34.9 microl, 0.432 mmol) were added to a solution of 17 (300 mg, 0.432 mmol) in dichloromethane (2 ml) at 0 ° C. The reaction mixture was stirred at this temperature for 2 h and then the solution was diluted with dichloromethane (15 ml) and washed with 0.1N hydrochloric acid (15 ml). The organic layer was dried over sodium sulfate, filtered and the solvent removed under reduced pressure to give 42 (318 mg, 100%) as a white solid, which was used in the subsequent reactions without further purification.

Rf: 0.5 (ethyl acetate: methanol 5: 1). 1 H NMR (300 MHz, CDCl ₃ ): delta 6.66 (s, 1H), 5.93 (d,> 1.2 Hz, 1H), 5.83 (d,> 1.2 Hz, 1H), 5.42 (t, J = 6.6 Hz, 1H ), 5.07 (d,> 5.7 Hz, 1H), 4.98 (d,> 5.7 Hz, 1H), 4.16 (d,> 1.8 Hz, 1H), 4.11 (d,> 2.7 Hz, 1H), 3.98 (bs. 1H), 3.73-3.61 (m, 2H), 3.64 (s, 3H), 3.52-3.48 (m, 1H), 3.50 (s, 3H), 3.33 (d,> 9.6 Hz, 1H), 3.17-3.14 ( m, 1H), 2.97-2.87 5 (m, 1H), 2.75-2.70 (d,> 16.8 Hz, 1H), 2.26 (s,

6H), 2.16 (s, 3H), 1.96 (s, 3H), 1.70 (dd, J = 11.7 Hz, J = 15.6 Hz, 1H), 1.33 (s, 6H), 0.59 (d,> 6.0 Hz, 3H ).

¹³ C NMR (75 MHz, CDCl ₃ ): delta 172.0, 10 168.3, 162.3, 148.2, 144.4, 140.4, 140.2, 130.9,

130.5, 125, 3, 123.4, 120.8, 117.6, 112.7, 111.7,

101.4, 99.1, 79.2, 59.5, 58.8, 57.5, 57.4, 56.4, 55.5,55.0,41.3,39.0,28.2,26.4,24.6,19.9,18.4,

15.4, 9.1.

ESI-MS m / z: Calcd. For C ₃₈ H ₄₉ N ₅ O ₁₀ : 735.82. Found (M + H) ⁺ : 736.3.

Example 26

To a solution of 42 (318 mg, 0.432 mmol) in dichloromethane (2.16 ml) was added trifluoroacetic acid (1.33 ml, 17.30 mmol) and the reaction mixture was stirred at 23 ° C for 3.5 h. The reaction mixture was fixed at 0 ° C with saturated aqueous sodium bicarbonate solution (60 ml) and extracted with dichloromethane (2x70 ml). The combined organic layers were dried over sodium sulfate and concentrated in vacuo. The residue was purified by flash column chromatography (silica, ethyl acetate: methanol 20: 1) to give 43 (154 mg, 60%) as a white solid.

Rf: 0.22 (ethyl acetate: methanol 5: 1). 1 H NMR (300 MHz, CDCl ₃ ): delta 6.47 (s, 1H), 6.22 (bs,

1H), 5.95 (d,> 1.2 Hz, 1H), 5.88 (d,> 1.2 Hz, 1H), 4.08-4.06 (m, 2H), 4.01 (bs, 1H), 3.69 (s, 3H), 3.49 ( d,> 3.6 Hz, 1H), 3.33 (d,> 8.1 Hz, 1H), 3.26-3.22 (m, 1H), 2.95 (dd, J = 8.1 Hz, J = 18 Hz, 1H), 2, 80- 2.76 (m, 2H), 2.58 (d,> 18 Hz, 1H), 2.29 (s, 3H), 2.27 (s, 3H), 2.21 (s, 3H), 1.96 (s, 3H), 1.77 (dd, J = 12.3 Hz, J = 15.6 Hz, 1H), 0.90 (d,> 6.9 Hz, 3H).

¹³ C NMR (75 MHz, CDCl ₃ ): delta 174.8, 169.0, 146.8, 144.4, 142.8, 140.5, 140.2, 131.1,

128.8, 120.8, 120.5, 117.1, 112.9, 111.6, 101.5, 60.3,59.0,56.5,56.3,55.6,55.1,50.2,41.6,39.5,

26.8, 26.3, 24.9, 20.2, 15.4, 9.2.

ESI-MS m / z: Calcd. For C ₃ H ₃₇ N ₃ O ₇ : 591.65. Found (M + H) ⁺ : 592.3.

65893 Bl

Example 27

44

To a solution of 43 (154 mg, 0.26 mmol) in dichloromethane (1.3 ml) was added phenylisothiocyanate (186 µl, 1.56 mmol) and the reaction mixture was stirred at 23 ° C for 2 h. The reaction mixture was concentrated in vacuo and the residue purified by flash column chromatography (silica, gradient hexane to hexane: ethyl acetate 1: 1) to give 44 (120 mg, 63%) as a white solid.

Rf: 0.41 (ethyl acetate: methanol 5: 1). 1 H NMR (300 MHz, CDCl ₃ ): delta 8.17 (s, 1H), 7.49-7.44 (m, 3H), 7.31-7.24 (m, 3H), 7.05 (d, J = 6.9 Hz, 1H), 5.98 ( d,> 1.2 Hz, 1H), 5.87 (d,> 1.2 Hz, 1H), 5.52 (bs, 1H), 4.54 (t,> 6.6 Hz, 1H), 4.15 (d,> 2.1 Hz, 1H), 4.03 (d,> 2.7 Hz, 2H), 3.80 (bs, 1H), 3.66 (s, 3H), 3.40 (bs, 1H), 3.32 (d,> 7.8 Hz, 1H), 3.16 (d,> 11.7 Hz. 1H), 2.82-2.61 (m, 3H), 2.29 (s, 3H), 2.20 (s, 3H), 2.01 (s, 3H), 1.99 (s, 3H), 1.80 (dd, 1, = 12.0 Hz. J = 15.9 Hz, 1H), 0.62 (d,> 6.0 Hz, 3H).

¹³ C NMR (75 MHz, CDCl ₃ ): delta 178.5, 171.9, 168.7, 146.7, 144.5, 142.6, 140.6, 140.3,

136.3, 131.0, 129.9, 128.9, 126.7, 124.4, 120.9, 120.6,117.7,116.6,112.7,111.9,101.4,60.4,58.7,

57.5,56.1,55.7,55.1,53.3,41.4,38.8,26.3,24.4,

20.2, 18.1, 15.3, 9.2.

ESI-MS m / z: Calculated for C „H ,,N, O, S:

726.3. Found (M + H) ⁺ : 727.3.

Example 28

NHCSNHPh

5.3N HCl and Dioxane

2.5b, 23 ° C

Me

Me

To a solution of 44 (120 mg, 0.165 mmol) in dioxane (0.9 ml) was added 5.3N hydrochloric acid / dioxane (1.8 ml) and the reaction mixture was stirred at 23 ° C for 2.5 h. Dichloromethane (10 ml) and water (5 ml) were then added to the reaction mixture and the organic layer was decanted. The aqueous phase was basified with saturated aqueous sodium bicarbonate solution (20 ml) (pH 8) at 0 ° C and then extracted with dichloromethane (2x15 ml). The combined organic extracts were dried over sodium sulfate and concentrated in vacuo to give 45 (75 mg, 87%) as a white solid, which was used in subsequent reactions without further purification.

Rf: 0.23 (ethyl acetate: methanol 5: 1). 1 H NMR (300 MHz, CDCl ₃ ): delta 6.43 (s, 1H), 5.94 (d,> 1.2 Hz, 1H), 5.87 (d,> 1.2 Hz, 1H), 4.10 (d,

65893 Bl> 2.1 Hz, 1H), 3.98 (d,> 2.4 Hz, 2H), 3.91 (bs, 1H), 3.69 (s, 3H), 3.34-3.25 (m, 2H), 3.05 (dd, J = 1.8) Hz, J = 8.1 Hz, 1H), 2.80-2.73 (m, 3H), 2.46 (d,> 18 Hz, 1H), 2.30 (s, 3H), 2.28 (s, 3H), 2.20 (s, 3H) , 1.98 (s, 3H), 1.79 (dd, J = 12.6 Hz, J = 16.2 Hz, 1H).

¹³ C NMR (75 MHz, CDCl ₃ ): delta 168.7,

146.7, 144.4, 142.9, 140.4, 130.4, 128.9, 121.1,

120.8,117.8,116.8,113.6,111.5,101.4,67.6,60.5,

59.8, 58.4, 56.6, 55.8,55.3,43.6,41.8, 31.3,25.6,

20.2, 15.6, 9.2.

ESI-MS m / z: Calculated for C _2g H ₃₂ N ₆ O ₆ :

520.58. Found (M + H) ⁺ : 521.3.

Example 29

To a solution of 45 (10 mg, 0.02 mmol) in dichloromethane (0.4 ml) was added phthalic anhydride (2.84 mg, 0.02 ramol) and the reaction mixture was stirred at 23 ° C for 2 h. Carbonyldiimidazole (0.5 mg, 0.003 mmol) was then added and the reaction mixture was stirred at 23 ° C for 7 h. Carbonyldiimidazole (2.61 mg, 0.016 mmol) was then added and the reaction mixture was stirred at 23 ° C for a further 17 h. The solution was diluted with dichloromethane (10 ml) and washed with 0.1 N hydrochloric acid (5 ml). The organic layer was dried over sodium sulfate, filtered and the solvent was removed under reduced pressure. The residue was purified by flash chromatography (RP-18, CH ₃ CN: H ₂ O 60:40) to give phthalacidin (11.7 mg, 93%) as a white solid.

Rf: 0.37 (CH ₃ CN: H ₂ O 7: 3, RP-18). ¹ H NMR (300 MHz, CDCl ₃ ): delta 7.72-7.68 (m, 2H), 7.677.63 (m, 2H), 6.38 (s, 1H), 5.69 (d,> 1.2 Hz, 1H), 5.64 (d ,> 1.2 Hz, 1H), 5.30 (bs, 1H), 4.254.21 (m, 2H), 4.02 (d,> 2.1 Hz, 1H), 3.64-3.62 (m, 5H), 3.33 (d,> 8.4 Hz, 1H), 3.21-3.16 (m, 1H), 3.02 (dd, J = 8.1 Hz, J = 18 Hz, 1H), 2.76 (dd, J = 1.8 Hz, J = 15.6 Hz, 1H), 2.63 ( d,> 17.7 Hz, 1H), 2.29 (s, 3H), 2.28 (s, 3H), 2.21 (s, 3H), 2.0 (s, 3H), 1.73 (dd, J = 12.0 Hz, J = 15.3 Hz , 1H).

¹³ C NMR (75 MHz, CDCl ₃ ): delta 168.5,

167.6, 146.2, 144.2, 142.5, 140.5, 133.4, 131.8,

130.7, 128.2, 120.9, 120.8, 117.9, 116.4, 113.6,

60.4, 60.0,57.0,56.3,55.6,55.4,41.6,41.5,26.5,

25.2, 20.2, 15.7, 9.4.

ESI-MS m / z: Calculated for C18H19N3O650.

34 4 8

Found (M + H) ⁺ : 651.2.

Example 30

65893 Bl

To a solution of 25 (18 mg, 0.032 mmol) in dimethylformamide (0.05 ml) at 0 ° C was added cat. dimethylaminopyridine (0.5 mg, 0.004 mmol), imidazole (5 mg, 0.08 mmol) and t-butyldiphenylsilyl chloride (12.5 microl, 0.048 mmol) and the reaction mixture was stirred at 23 ° C for 6 h. Water (10 ml) at 0 ° C was added and the aqueous phase was extracted with hexane: ethyl acetate 1:10 (2x10 ml). The organic layer was dried over sodium sulfate, filtered and the solvent was removed under reduced pressure. The residue was purified by flash chromatography (silica, hexane: ethyl acetate 3: 1) to give 26 (27 mg, 88%) as a white solid.,

Rf: 0.29 (hexane: ethyl acetate 3: 1). 1 H NMR (300 MHz, CDCl ₃ ): delta 7.61-7.58 (m, 2H), 7.427.28 (m, 8H), 6.71 (s, 1H), 6.19-6.02 (m, 1H).

5.78 (d, J = 1.2 Hz, 1H), 5.64 (d, J = 1.2 Hz, 1H), 5.40 (dd, Jl = 1.2 Hz, J = 17.1 Hz, 1H), 5.27 (dd, J = 1.2Hz, J = 10.2 Hz, 1H), 5.13 (s, 2H), 4.45 (d, J = 2.4 Hz, 1H), 4.24 (d, J = 2.1 Hz, 1H), 4.17-4.06 (m, 3H), 3.75 ( s, 3H), 3.64 (dd, J = 2.4 Hz, J = 9.9 Hz, 1H), 3.59 (s, 3H), 3.42-3.21 (m, 4H), 3.10 (dd, J = 8.1 Hz, J = 17.7 Hz, 1H), 2.70 (d, J = 17.7 Hz, 1H), 2.33 (s, 3H), 2.26 (s, 3H), 2.11 (s, 3H), 2.08-1.89 (m, 1H), 0.87 (s , 9H).

¹³ CNMR (75 MHz, CDCl3): delta 148.5,

148.3, 148.1, 144.0, 139.0, 135.6, 135.4, 133.8,

133.1, 132.6, 130.5, 130.3, 129.6, 129.4, 127.5,

127.4, 125.1, 124.3, 121.6, 118.5, 117.5, 112.9,

111.7,100.8,99.2,74.0,67.7,61.5,59.6,59.0,57.7,

57.1, 55.4, 41.6, 29.6, 26.6, 25.5, 18.8, 15.8, 9.2.

FSI-MS m / z: Calculated for C ₄₇ H ₅₅ N ₃ O ₇ Si:

801.3. Found (M + H) ⁺ : 802.3.

Example 31

Me

BujSnH, (PPhjljPdClj

AcOH, CH ₂ CI ₂ , 1h. 25 ° C

MOMO

OH

Me

CN OTBDPS

ET-11

To a solution of 26 (7 mg, 0.0087 mmol) in dichloromethane (0.15 ml) at 23 ° C was added acetic acid (2.5 microl, 0.044 mmol), (PPh ₃ ) 2PdCl ₂ (0.5 mg, 6.96 x 10 · ⁴ mmol) and Bu ₃ SnH (3.5 microl, 0.013 mmol). The reaction mixture was stirred at this temperature for 1 h. The solution was diluted with a mixture of hexane: ethyl acetate 5: 1 (0.5 ml) and poured into a pressure column (silica, gradient hexane: ethyl acetate from 5: 1 to 1: 1) to give ET-11 (5 mg, 75%) as a white solid.

Rf: 0.36 (hexane: ethyl acetate 1: 5, silica). Ή NMR (300 MHz, CDC1 _3): delta 7.56 (m, 2H), 7.41-7.25 (m, 8H), 6.67 (s, IH), 5.72 (d, J = 1.0 Hz, IH), 5.58 (d, J = 1.0 Hz, 1H), 5.51 (s, 1H), 5.38 (d, J = 5.75 Hz, 1H), 5.16 (d, J = 5.7 Hz,

1H), 4.57 (d, J = 2.9 Hz, 1H), 4.21 (m, 1H), 4.09 (m, 1H), 3.72 (s, 3H), 3.71 (s, 3H), 3.68 (dd, J = 2.1). Hz, J = 10.4Hz, 1H), 3.38-3.26 (m, 3H), 3.11 (dd, J = 2.5 Hz, J = 15.7 Hz, 1H), 3.01 (dd, J = 8.9 Hz, J = 17.9 Hz. 1H), 2.70 (d, J = 17.9 Hz, 1H), 2.31 (s, 3H), 2.25 (s, 3H), 2.06 (s, 3H), 1.89 (dd, J = 12.1 Hz, J = 15.7 Hz. 1H), 0.9 (s, 9H).

¹³ C NMR (75 MHz, CDCl ₃ ): delta 149.0,

147.4, 145.3, 144.3, 136.3, 135.7, 135.4, 133.2, 130.9, 130.5, 129.6, 129.5, 127.5, 125.0, 118.6,

112.5,112.1, 105.7, 100.5, 99.8, 68.5, 61.5, 59.7,

58.8, 57.7, 56.9,56.5,55.4,41.7,26.6,26.2,25.5,

18.9, 15.8, 14.2, 8.7.

ESI-MS m / z: Calcd. For C ₄₄ H ₅₁ N ₃ O ₇ Si: 761. Found (M + H) ⁺ : 762.

65893 Bl

Example 32

A solution of 2 (3.0 g, 5.46 mmol) and phenylisothiocyanate (3.92 ml, 32.76 mmol) in dichloromethane (27 ml) was stirred at 23 ° C for 1.5 h. The reaction mixture was partitioned into dichloromethane (10 ml) and water (5 ml). The organic layer was dried over sodium sulfate, filtered and concentrated. The residue was purified by flash chromatography (silica, gradient hexane to 2: 3 hexane: ethyl acetate) to give 3 (3.29 g, 88%) as a yellow solid.

Rf: 0.27 (CA3: 2, RP-C18). 1 H NMR (300 MHz, CDCl ₃ ): delta 7.77 (bs, 3H), 7.41-7.11 (m, 5H), 6.65 (d, 1H), 6.29 (s, 1H), 5.6-5.5 (m, 1H). 4.19-4.14 (m, 2H), 4.08 (d, 1H), 3.92 (s, 3H), 3.87-3.65 (m, 6H), 3.77 (s, 3H), 3.37-2.98 (m, 8H), 2.50 ( d, 1H), 2.31 (s, 3H), 2.20 (s, 3H), 1.96 (d, 1H), 1.87 (s, 3H), 1.81-1.75 (m, 1H), 0.96 (d, 3H).

^I3 CNMR (75 MHz, CDCL): delta 185.7,180, 9,178.9,172.0,155.7,147.1,143.2,142.4,136.0,

135.1, 130.5, 129.9, 129.3, 128.5, 126.9, 124.4, 120.2,117.4,116.3,77.1,60.9,58.6,56.2,55.8,55.0,

54.6, 53.5, 41.7, 40.3, 25.1, 24.5, 18.4, 15.8, 8.7.

ESI-MS m / z: Calculated for C, H, N, O, S:

40 6 6

684.8. Found (M + H) ⁺ : 685.2.

Example 33

A solution of 3 (0.143 g, 0.208 ml) in 6.5M hydrochloric acid / dioxane (150 ml) was stirred at 23 ° C for 6 h. Toluene (3 ml) was then added to the reaction mixture and the organic layer was decanted. The residue was partitioned between saturated aqueous sodium bicarbonate solution (3 ml) and chloroform (3x3 ml). The organic layers were dried and concentrated to give the title compound as a mixture of 4 and 6 (4: 6 90:10), which slowly cyclized to 6 on standing.

Rf: 0.4 (ethyl acetate: methanol 5: 1, silica ⁴⁰ ). Ή NMR (300 MHz, CDCl ₃ ): δ 6.45 (s, 1H), 4.16 (m, 1H), 4.02 (d, 1H), 3.96 (s, 3H), 3.79 (m, 2H), 3.75 (s, 3H), 3.35 (m, 1H), 3,203.00 (m, 3H), 2.87 (d, 1H), 2.75 (d, 1H), 2.43 (d, 1H), 2.34 (s, 3H), 2.30 (s , 3H), 1.93 (s, 3H), 1.72 ⁴⁵ 1.5 (m, 3H).

ESI-MS m / z; Calculated for C, H, .N.O ·

478.5. Found (M + H) ⁺ : 479.2.

65893 Bl

Example 34

A solution of 3 (0.143 g, 0.208 mmol) in 6.5M hydrochloric acid / dioxane (150 ml) was stirred at 23 ° C for 1 h. Evaporation of the solvent gave a residue which was purified by flash chromatography (ethyl acetate / methanol / triethylamine 100: 25: 0.1) to give 6 (80 mg, 83%) as a yellow solid.

Rf: 0.26 (ACOD 3: 2, RP-C18). Ή NMR (500 MHz, CDCl ₃ ): δ 6.46 (s, 1H), 5.09 (bs, 1H), 4.67 (dd, J = 8.3Hz, J = 7.8Hz, 1H), 4.24 (d.

1H), 4.16 (s, 3H), 3.93 (d, J = 2.7 Hz, 1H), 3.8 (m, 2H), 3.77 (s, 3H), 3.45 (m, 2H), 3.08 (dd, J = 17.9 Hz, J = 3.6 Hz, 1H), 2.78 (m, 1H), 2.55 (d, 1H), 2.3 (m, 1H), 2.3 (s, 3H), 2.28 (s, 3H), 1.90 (s, 3H) ¹³ C NMR (75 MHz, CDCl ₃ ): delta 186.2,

162.1, 154.9, 146.9, 145.3, 143.0, 130.1, 129.4, 128.1,125.0,121.4,116.4,116.2,66.6,60.7,60.7,60.1,

59.6, 58.8,55.6, 54.9, 41.9, 25.3, 24.7, 15.7, 8.9.

ESI-MS m / z: Calculated for C ₂₆ H ₂₈ N ₄ O ₄ :

460.5. Found (M + H) ⁺ : 461.1.

Example 35

To a solution of 3 (2.38 g, 3.47 mmol) in dioxane (5 ml) was added 5.3 M hydrochloric acid in dioxane (34 ml) and the reaction mixture was stirred at 23 ° C for 45 min. Acetic anhydride (51 ml, 539.5 mmol) was then added and the reaction mixture was stirred for 4 h. The reaction mixture was cooled to 0 ° C and partitioned between aqueous saturated sodium carbonate solution (300 ml) and ethyl acetate (300 ml) at this temperature. The organic phase was dried over sodium sulfate, filtered and concentrated. The residue was purified by flash chromatography (silica, gradient dichloromethane to dichloromethane: ethyl acetate 1: 2) to give 5 (1.75 g, 97%) as a yellow solid.

Rf: 0.53 (ACMW 3: 2, RP-C18). 1 H NMR ³⁵ (300 MHz, CDCl ₃ ): delta 6.51 (s, 1H), 5.98 (bs, 1H), 4.84 (dd, 1H), 4.17 (d, 1H), 4.00 (d, 1H), 3.99 (s , 3H), 3.85 (bs, 1H), 3.81 (m, 1H), 3.74 (s, 3H), 3.70 (d, 1H), 3.23 (m, 1H), 3.11 (dd, 1H), 3.09 (m, 1H), 2.93 (m, 2H), 2.44 (d, 1H), 2.67 (s, ⁴⁰ 3H), 2.25 (s, 3H), 1.70 (s, 3H), 1.60-1.50 (m, 2H), 1.29 ( s, 3H).

¹³ C NMR (75 MHz, CDCl ₃ ): delta 185.9,

180.8, 169.9, 160.2, 156.2, 147.0, 143.1, 140.4,

136.1, 130.6, 129.6, 127.9, 120.4, 117.2, 61.0, ⁴⁵ 60.7, 58.6, 56.1,55.7,55.1, 54.3,41.8,41.1,25.7,

23.9, 22.2, 15.7, 8.7.

ESI-MS m / z: Calculated for C ₂₈ H ₃₂ N ₄ O ₆ :

520.6. Found (M + H) ⁺ : 521.1.

65893 Bl

Example 36

To a solution of 5 (1.75 g, 3.36 mmol) in dichloromethane (17 ml) at 0 ° C was added diisopropylethylamine (11.71 ml, 67.23 mmol), dimethylaminopyridine (20 mg, 0.17 mmol) and bromomethyl ether (4.11 ml, 50.42 mmol). . After 6 h at 23 ° C, the reaction mixture was partitioned between aqueous saturated sodium bicarbonate solution (25 ml) and dichloromethane (50 ml). The organic layer was dried over sodium sulfate and the solvent was removed under reduced pressure. The crude product was purified by flash chromatography (RP-18, CH ₃ CI / water 1: 1) to give 7 (1.32 g, 70%) as a yellow solid.

Rf: 0.34 (ACOD 2: 3, RP-C18). NMR (300 MHz, CDCl3): delta 6.74 (s, 1H), 5.14 (s, 2H), 4.82 (m, 1H), 4.22 (d, 1H), 4.00 (s, 3H), 4.0 ₁₅ (m, 1H), 3.83 (m, 2H), 3.7 (s, 3H), 3.58 (s, 3H),

3.4 (m, 1H), 3.2-2.95 (m, 6H), 2.43 (d, 1H), 2.37 (s, 3H), 2.22 (s, 3H), 1.890 (s, 3H), 1.5-1.4 (m. 2H), 1.31 (s, 3H).

¹³ C NMR (75 MHz, CDCl ₃ ): delta 185.9, ₂₀ 180.7, 169.6, 156.2, 148.9, 148.5, 140.3, 136.2,

131.3, 130.1, 127.7, 124.6, 123.7, 117.3, 99.5, 99.2,60.9,59.7,58.8,57.7,56.4,55.7,55.0,54.2, 51.0,41.6,41.0,40.5,25.5,23.9,22.3,19.3,15.6,

14.6, 8.6.

ESI-MS m / z: Calculated for C ₃₀ H ₃₆ N ₄ O ₇ :

520.6. Found (M + H) ⁺ : 565.3.

Example 37

OMe

To a solution of 7 (0.37 g, 0.65 mmol) in methanol (74 ml) at 0 ° C was added a 1M solution of sodium hydroxide (130 ml). The reaction mixture was stirred for 15 min and then fixed at 0 ° C with 6M hydrochloric acid to pH 5. The reaction mixture was extracted with ethyl acetate (3 x 50 ml) and the combined organic layers dried over sodium sulfate and concentrated. under vacuum. The residue was purified by flash chromatography (RP-18, CH ₃ CN / water 1: 1) to give 8 (232 mg, 65%) as a yellow oil.

Rf: 0.5 (ACI: water 3: 2, RP-C18). NMR (300 MHz, CDCl ₃ ): delta 6.75 (s, 1H), 5.15 (s, 2H), 4.86 (m, 1H), 4.26 (d, 1H), 4.01 (d, 1H), 40 3.88-3.81 (m, 2H), 3.70 (s, 3H), 3.58 (s, 3H), 3.39 (m, 1H), 3.27-3.21 (m, 1H), 3.18-3.08 (m, 2H), 3.032.97 (m , 1H), 2.47 (d, 1H), 2.37 (s, 3H), 2.22 (s, 3H), 1.90 (s, 3H), 1.57-1.46 (m, 2H), 1.33 (s, 3H).

¹³ C NMR (75 MHz, CDCl ₃ ): delta 185.3, 45 180.6, 175.9, 170.1, 151.5, 148.9, 148.6, 143.3,

133.7, 131.5, 129.9, 124.7, 123.5, 117.1, 117.0,

99.2, 59.8,58.7,57.8,56.3,55.3,54.9,54.3,41.5,

40.7, 29.6, 25.5, 24.4, 22.2, 20.7, 15.7, 8.0.

ESI-MS m / z: Calculated for C ₂₉ H ₃₄ N ₄ O ₇ :

550.6. Found (M + H) ⁺ : 551.2.

65893 Bl

To a degassed solution of compound 8 (240 mg, 0.435 mmol) in dimethylformamide (30 ml) was added 10% palladium on carbon (48 mg) and the reaction mixture was stirred under hydrogen (atmospheric pressure) for 1 hour. The reaction mixture was filtered through a pad of selite under argon in a Sehlenk tube as a colorless solution containing anhydrous cesium carbonate (240 mg, 0.739 mmol). Bromochloromethane (0.566 ml, 8.71 mmol) was then added. The tube was sealed and stirred at 90 ° C for 3 h. The reaction mixture was cooled and filtered through Celite and washed with dichloromethane. The organic layer was concentrated and dried over sodium sulfate to give 9 as a brown oil, which was used in the next step without further purification.

Rf: 0.36 (hexane: ethyl acetate 1: 5, silica). NMR (300 MHz, CDCl ₃ ): delta 6.71 (s, 3H), 5.89 (d, 1H), 5.81 (d, 1H), 5.63 (bs, 1H).

5.33 (d, 1H), 5.17 (d, 1H), 4.97 (m, 1H), 4.20 (d,

1H), 4.09 (m, 1H), 3.99 (m, 1H), 3.68 (m, 1H), 3.65 (s, 6H), 3.59-3.47 (m, 4H), 3.37-3.27 (m, 2H).

3.14-2.97 (m, 2H), 2.62 (d, 1H), 2.32 (s, 3H), 2.20 (s, 3H), 2.08 (s, 3H), 1.72 (m, 1H), 1.36 (s, 3H) .

¹³ C NMR (75 MHz, CDCl ₃ ): delta 169.8,

149.1, 147.4, 145.3, 136.2, 130.9, 130.8, 125.0,

122.9, 117.7, 112.6, 111.8, 106.4, 100.8, 99.8,

59.8, 58.9,57.7,56.6,56.4,55.5,55.2,41,6,40.1,

29.6, 25.9, 25.0, 22.6, 15.6, 8.8.

ESI-MS m / z: Calculated for C, _n H, NO, Si:

36 4 7

564.6. Found (M + H) ⁺ : 563.3.

Example 39

To a flask containing 9 (245 mg, 0.435 mmol) in dimethylformamide (4 ml) at 0 ° C was added cesium carbonate (425 mg, 1.30 mmol) and allyl bromide (376 microl, 4.35 mmol) and the reaction mixture was stirred at 23 ° C. for 1 h. The reaction mixture was filtered through a pad of celite and partitioned between dichloromethane (25 ml) and water (10 ml). The organic phase was dried over sodium sulfate and concentrated under reduced pressure to give a residue which was purified by flash chromatography (silica, chloroform: ethyl acetate 1: 2) to give 10 (113 mg, 43%) as yellow oil.

Rf: 0.36 (hexane: ethyl acetate 1: 5). 1 H NMR (300 MHz, CDCl ₃ ): delta 6.74 (s, 1H), 6.3-6.0 (m, 1H), 5.94 (d, 1H), 5.87 (d, 1H), 5.43-5.36 (m, 2H). 5.22 (s, 2H), 5.00 (m, 1H), 4.22 (m, 1H), 4.174.01 (m, 1H), 3.98 (m, 2H), 3.71-3.67 (m, 1H), 3.69 (s. 6H), 3.62-3.51 (m, 3H), 3.58 (s, 3H), 3.393.37 (m, 1H), 3.31-3.26 (m, 3H), 3.09 (dd, 1H), 2.56 (d, 1H) , 2.36 (s, 3H), 2.21 (s, 3H), 2.11 (s,

65893 Bl

3H), 2.24-2.10 (m, 1H), 1.82-1.73 (m, 1H), 1.24 (bs, 3H).

^{, 3} C NMR (75 MHz, CDCl 3): delta 169.4,

148.8, 148.3, 139.1, 133.7, 130.9, 130.3, 125.2,

120.2,117.7,113.1,112.6,101.3,99.3,74.1,59.7,

59.3,57.8,57.0,56.1,56.1,55.2,41.6,41.0,40.9,

29.7, 26.3, 22.5, 15.6, 9.3.

ESI-MS m / z: Calculated for C ₃₃ H ₄₀ N ₄ O ₇ :

604.7. Found (M + H) ⁺ : 605.3.

Example 40

(In SA

Acetyl chloride (2.79 microl, 0.039 mmol) and pyridine (3.2ml, 0.039 mmol) were added to a solution of 9 (22 mg, 0.039 mmol) in dichloromethane (0.2 ml) at 0 ° C. The reaction mixture was stirred for 1 h and then the solution was diluted with dichloromethane (10 ml) and washed with 0.1N hydrochloric acid (5 ml). The organic layer was dried over sodium sulfate, filtered and the solvent removed under reduced pressure to give 46 (22 mg, 93%) as a white solid.

Rf: 0.4 (hexane: ethyl acetate 1: 5). Ή NMR (300 MHz, CDCl ₃ ): delta 6.74 (s, 1H), 5.97 (d,> 0.9 Hz, 1H), 5.91 (d, J = 0.9 Hz, 1H), 5.12 (d, ²⁰ > 5.7 Hz. 2H), 5.04 (d,> 5.7 Hz, 1H), 4.90 (t,> 6 Hz, 1H), 4.17 (d,> 2.7 Hz, 1H), 4.05 (d,> 2.7

Hz, 1H), 4.01 (bs, 1H), 3.71 (s, 3H), 3.57 (s, 3H), 3.50-3.44 (m, 2H), 3.38-3.36 (m, 1H), 3.30-3.26 (m. 1H), 3.00 (ddm J = 7.8, Hz, J = 18.0 Hz, 1H), ²⁵ 2.79 (d,> 12.9 Hz, 1H), 2.60 (d,> 18.0 Hz, 1H).

2.35 (s, 3H), 2.32 (s, 3H), 2.21 (s, 3H), 2.00 (s, 3H), 1.68 (dd, J = 11.7 Hz, J = 15.6 Hz, 1H).

ESI-MS m / z: Calculated for C ₃₂ H ₃₈ N ₄ O _g :

606.67. Found (M + H) ⁺ : 607.3.

Example 41

oA

Me with SA

To a solution of 46 (8 mg, 0.013 mmol) in dioxane (0.1 ml) was added 5.3N hydrochloric acid / dioxane (0.5 ml) and the reaction mixture was stirred at 23 ° C for 1 h. The solution was then diluted with dichloromethane (5 ml) and washed with 0.1N hydrochloric acid (3 ml). The organic layer was dried over sodium sulfate, filtered and the solvent removed under reduced pressure to give 47 (5 mg, 70%) as a white solid.

Rf: 0.4 (hexane: ethyl acetate 1: 5). Ή NMR (300 MHz, CDC1 _3): delta 6.51 (s, IH), 5.97 (d,> 1.2Hz, IH), 5.91 (d,> 1.2Hz, IH), 4.97 (bs, IH), 4.11 (bs , 1H), 4.04-4.02 (m, 2H), 3.75 (s,

65893 Bl

3H), 3.65 (d, > 2.1 Hz, 2H), 3.56-3.30 (m, 2H),

3.04 (dd, J = 7.5 Hz, J = 18.0 Hz, 1H), 2.80 (d,> 14.4 Hz, 1H), 2.59 (d,> 18.3 Hz, 1H), 2.33 (s,

3H), 2.24 (s, 3H), 2.00 (s, 3H), 1.76 (dd, J = 12.0)

Hz, J = 15.9 Hz, 1H), 1.33 (s, 3H), 1.25 (s, 3H).

ESI-MS m / z: Calculated for C ₃₀ H ₃₄ N ₄ O _7: 562.61. Found (M + H) ⁺ : 563.3.

Example 42

To a solution of 45 (10 mg, 0.0192 mmol) in dichloromethane (0.3 ml) at 0 ° C was added isovaleryl chloride (2.34 microl, 0.0192 mmol) and pyridine (1.55 ml, 0.0192 mmol). The reaction mixture was stirred for 1 h and then the solution was diluted with dichloromethane (5 ml) and washed with 0.1 N hydrochloric acid (3 ml). The organic layer was dried over sodium sulfate, filtered and the solvent was removed under reduced pressure. The residue was purified by flash column chromatography (silica, hexane: ethyl acetate 1: 2) to give 48 (11 mg, 95%) as a white solid.

Rf: 0.12 (hexane: ethyl acetate 1: 2). Ή NMR (300 MHz, CDC1 _3): delta 6.50 (s, IH), 5.98 (d,> 1.5 Hz, IH), 5.91 (d,> 1.5 Hz. IH), 5.75 (s, IH), 5.02 (t ,> 5.4 Hz, 1H), 4.10 (d,> 1.5 Hz, 1H), 4.06 (d,> 2.7 Hz, 1H), 4.02 (d,> 2.7 Hz, 1H), 3.77 (s, 3H), 3.76- 3.71 (m, 1H), 3.86-3.28 (m, 3H), 3.04 (dd, 1, = 8.1 Hz, J = 18.3 Hz, 1H), 2.78 (d,> 15.9 Hz, 1H), 2.55 (d, > 18 Hz. 1H), 2.32 (s, 6H), 2.26 (s, 3H), 1.98 (s, 3H), 1.84-1.68 (m, 2H), 1.36 (d,> 7.2 Hz, 2H), 0.69 (d ,> 6.6 Hz, 3H), 0.62 (d,> 6.6 Hz, 3H).

ESI-MS m / z: Calculated for C ₃₃ H ₄₀ N ₄ O _? :

604.69. Found (M + H) ⁺ : 605.3.

Example 43

Me

To a solution of 45 (10 mg, 0.0192 mmol) in dichloromethane (0.3 ml) at 0 ° C was added isovaleryl chloride (2.34 microl, 0.0192 mmol) and pyridine (1.55 microl, 0.0192 mmol). The reaction mixture was stirred for 1 h and then the solution was diluted with dichloromethane (5 ml) and washed with 0.1 N hydrochloric acid (3 ml). The organic layer was dried over sodium sulfate, filtered and the solvent was removed under reduced pressure. The residue was purified by flash column chromatography (silica, hexane: ethyl acetate 1: 2) to give 49 (12.4 mg, 96%) as a white solid.

Rf: 0.7 (ethyl acetate: methanol 10: 1). 1 H NMR (300 MHz, CDCl ₃ ): delta 6.50 (s, 1H), 5.98 (d,> 1.5 Hz, 1H), 5.91 (d,> 1.5 Hz, 1H), 5.73 (s, 1H), 5.08 (t ,> 5.4 Hz, 1H), 4.10 (d,> 1.5 Hz,

65893 Bl

1H), 4.05 (m, 1H), 4.01 (tn, 1H), 3.76 (s, 3H), 3.65-3.61 (m, 1H), 3.40-3.27 (m, 3H), 3.03 (dd, J = 8.1 Hz) , J = 18.6 Hz, 1H), 2.78 (d,> 13.2 Hz,

1H), 2.57 (d,> 18.3 Hz, 1H), 2.32 (s, 6H), 2.31 (s, 3H), 2.25 (s, 3H), 1.99 (s, 3H), 1.79 (dd, J = 12.0

Hz, J = 16.5 Hz, 1H), 1.73-1.42 (m, 4H), 1.33-1.18 (m, 10H), 1.03 (m, 2H), 0.87 (t,> 6.6 Hz, 3H).

ESI-MS m / z: Calculated for C ₃₈ H ₅₀ N ₄ O ₇ : 674.83. Found (M + H) ⁺ : 675.5.

Example 44

To a solution of 45 (14.5 mg, 0.0278 mmol) in dichloromethane (0.3 ml) at 0 ° C was added trans-3-trifluoromethylsinamoyl chloride (4.76 microl, 0.0278 mmol) and pyridine (2.25 microl, 0.0278 mmol). The reaction mixture was stirred for 1 h and then the solution was diluted with dichloromethane (5 ml) and washed with 0.1 N hydrochloric acid (3 ml). The organic layer was dried over sodium sulfate, filtered and the solvent was removed under reduced pressure. The residue was purified by flash column chromatography (silica, hexane: ethyl acetate 1: 1) to give 50 (18.7 (d,> 16.0 Hz, 1H), 6.34 (s, 1H), 6.12 (d,> 16.0 Hz, 1H), 6.07 (d,> 0.9 Hz, 1H), 5.96 (d,> 0.9 Hz, 1H), 4.39 (d,> 2.4 Hz, 1H), 4.07-4.05 (m, ₂₀ 1H), 3.81 (bs. 1H), 3.46-3.51 (m, 3H), 3.42 (s, 3H), 3.09 (brd,> 12.0 Hz, 1H), 2.94-2.85 (m, 2H), 2.74 (d,> 18.3 Hz, 1H). 2.38 (s, 3H), 2.23 (s, 3H), 2.02 (s, 3H), 1.80 (s, 3H), 1.84-1.75 (m, 1H).

¹³ C NMR (75 MHz, CDCl ₃ ): delta 168.7, ₂₅ 165.3, 146.5, 144.7, 142.6, 140.6, 138.0, 135.9, 131.0, 130 $, 129.1, 128.6, 125.8, 125.7, 124.5,

124.4, 122.7, 121.2, 117.8, 116.5, 113.0, 112.0,

101.7, 60.4, 59.1, 56.5, 56.4, 55.6, 55.3, 41.8,

40.3, 26.6, 25.1, 20.3, 15.4, 9.3.

mg, 94%) as a white solid.

Rf: 0.64 (ethyl acetate: methanol 5: 1). Ή NMR

ESI-MS m / z: Calculated for C ₃₈ H ₅₀ N ₄ O ₇ :

674.83. Found (M + H) ⁺ : 675.5.

To a solution of 43 (33 mg, 0.0557 mmol) in dichloromethane (0.4 ml) at 0 ° C was added isovaleryl chloride (6.79 ml, 0.0557 mmol) and pyridine (4.5 ml, 0.0557 mmol). The reaction mixture was stirred for 1 h and then the solution was diluted with dichloromethane (5 ml) and washed with 0.1 N hydrochloric acid (3 ml). The organic layer was dried over sodium sulfate, filtered and the solvent was removed under reduced pressure. The residue was purified by flash column chromatography (silica, hexane: ethyl acetate 1: 2) to give more

65893 Bleaching 51 (34 mg, 91%) as a white solid.

Rf: 0.09 (hexane: ethyl acetate 1: 2). 1 H NMR (300 MHz, CDCl ₃ ): delta 6.46 (s, 1H), 6.10 (bs, 1H), 5.99 (d,> 0.9 Hz, 1H), 5.90 (d,> 0.9 Hz, 1H), 5.30 (t ,> 6.0 Hz, 1H), 4.10-4.05 (m, 3H), 3.81 (bs, 3H), 3.74 (s, 3H), 3.54 (bs, 1H), 3.38-

3.36 (m, 1H), 3.29-3.21 (m, 1H), 3.00 (dd, J = 8.0 Hz, J = 18.0 Hz, 1H), 2.25 (s, 3H), 2.20 (s, 3H), 2.00 (s , 3H), 1.95-1.90 (m, 3H), 0.87 (d, J = 6.6 Hz, 6H), 0.76 (d,> 6.0 Hz, 3H).

ESI-MS m / z: Calculated for C ₃₆ H ₄₅ N ₅ O _g :

675.77. Found (M + H) ⁺ : 676.3.

Example 46

To a solution of 43 (33 mg, 0.0557 mmol) in dichloromethane (0.4 ml) at 0 ° C was added trans-3-trifluoromethylsinamoyl chloride (9.52 microl, 0.0557 mmol) and pyridine (4.5 microl, 0.0557 mmol). The reaction mixture was stirred for 1 h and then the solution was diluted with dichloromethane (5 ml) and washed with 0.1 N hydrochloric acid (3 ml). The organic layer was dried over sodium sulfate, filtered and the solvent was removed under reduced pressure. The residue was purified by flash column chromatography (silica, hexane: ethyl acetate 1: 2) to give 52 (40 mg, 92%) as a white solid.

Rf: 0.21 (hexane: ethyl acetate 1: 2). 1 H NMR (300 MHz, CDCl ₃ ): delta 7.74-7.47 (m, 4H), 6.49 (s, 1H), 6.40 (d,> 15.6 Hz, 1H), 6.00 (d,> 1.5

Hz, 1H), 5.90 (d,> 1.5 Hz, 1H), 5.47 (t,> 6.0 Hz, 1H), 4.12-4.09 (m, 3H), 3.93 (bs, 1H), 3.71 (s, 3H). 3.59-3.58 (m, 1H), 3.38 (d,> 7.8 Hz, 1H), 3.29 (d,> 12.0 Hz, 1H), 3.00 (dd, J = 8.1 Hz, 25 J = 18.3Hz, 1H), 2.79 -2.78 (m, 1H), 2.65 (d, > 18.3

Hz, 1H), 2.29 (s, 6H), 2.28 (s, 3H), 2.22 (s, 3H), 1.84-1.80 (m, 1H), 0.85-0.84 (m, 3H).

¹³ C NMR (75 MHz, CDCl 3): delta 171.9,

168.8, 164.4, 146.9, 144.6, 143.0, 140.5, 140.5, 30 139.3, 135.7, 131.1, 131.0, 129.4, 129.1, 126.0,

124.1, 124.0, 122.4, 121.1, 120.7, 120.6, 117.7,

116.9, 112.8, 112.0,101.6, 60.6, 59.3, 57.1, 56.3, 55.9,55.2,49.0,41.7,39.9,26.5,25.1,20.2,18.4, 15.7, 9.3.

ESI-MS m / z: Calculated for C ₄₁ H ₄₂ FjN ₅ O ₈ :

789.8. Found (M + H) ⁺ : 790.3.

Example 47

65893 Bl

To a solution of 43 (10 mg, 0.0169 mmol) in dichloromethane (0.2 ml) at 23 ° C was added trifluoroacetic anhydride (2.38 microl, 0.0169 mmol). The reaction mixture was stirred for 5 h and then the solution was diluted with dichloromethane (5 ml) and washed with 0.1 N hydrochloric acid (3 ml). The organic layer was dried over sodium sulfate, filtered and the solvent was removed under reduced pressure. The residue was purified by flash column chromatography (silica, hexane: ethyl acetate 3: 2) to give 55 (10.7 mg, 93%) as a white solid.

Rf: 0.57 (ethyl acetate: methanol 5: 1). 1 H NMR (300 MHz, CDCl ₃ ): delta 6.45 (s, 1H), 6.00 (d,> 1.2 Hz, 1H), 5.90 (d,> 1.2 Hz, 1H), 5.87 (bs,

1H), 5.32 (bs, 1H), 4.12 (d,> 2.1 Hz, 1H), 4.08 (d,> 1.8 Hz, 1H), 3.78-3.56 (m, 3H), 3.72 (s, 3H), 3.40 ( d,> 8.1 Hz, 1H), 3.25 (d,> 9.3 Hz, 1H), 3.00 (dd, J = 8.4 Hz, J = 18.0 Hz, 1H), 2.77 (dd, J = 2.1 Hz, J = 15.9 Hz , 1H), 2.68 (d,> 18.6 Hz, 1H), 2.30 (s, 3H), 2.28 (s, 3H), 2.22 (s, 3H), 2.00 (s, 3H), 1.75 (dd, J = 11.4 Hz, J = 15.9 Hz, 1H), 0.69 (d,> 6.3 Hz, 3H).

¹³ C NMR (75 MHz, CDCl ₃ ): delta 170.1,

168.6, 156.0, 147.0, 144.6, 143.0, 140.6, 140.4, 131.0, 129.4, 120.9, 120.7, 117.6, 116.8, 112.4,

112.1, 101.6, 60.5, 59.0, 57.1, 56.3, 55.6, 55.2, 48.7, 41.6, 39.4, 26.5, 24.9, 20.2, 17.8, 15.4, 9.2.

ESI-MS m / z: Calculated for C ₃₃ H ₃₆ F ₃ N ₅ O ₈ : 15 687.63. Found (M + H) ⁺ : 688.66.

Example 48

To a solution of 19 (11 mg, 0.0169 mmol) in dichloromethane (0.2 ml) at 23 ° C was added trifluoroacetic anhydride (2.38 microl, 0.0169 mmol). The reaction mixture was stirred for 5 h and then the solution was diluted with dichloromethane (5 ml) and washed with 0.1 N hydrochloric acid (3 ml). The organic layer was dried over sodium sulfate, filtered and the solvent was removed under reduced pressure. The residue was purified by flash column chromatography (silica, hexane: ethyl acetate 3: 2) to give 54 (10.7 mg, 93%) as a white solid.

Rf: 0.6 (ethyl acetate: methanol 5: 1). 1 H NMR (300 MHz, CDCl ₃ ): delta 7.33 (d,> 6.3 Hz, 1H),

6.04 (m, 1H), 5.95 (d,> 1.5 Hz, 1H), 5.84 (d, ₃₀ > 1.5 Hz, 1H), 5.32 (m, 2H), 5.21 (m, 1H), 4.11 (m, 4H) , 3.73 (s, 3H), 3.64 (m, 2H), 3.51 (m, 1H),

3.37 (d,> 7.8 Hz, 1H), 3.22 (m, 2H), 3.03 (dd, J = 8.1 Hz, J = 18.3Hz, 1H), 2.60 (d,> 18.3Hz, 1H), 2.29 (s. 3H), 2.24 (s, 3H), 2.08 (s, 3H), 1.86 (dd, J = 12Hz, J = 16.2Hz, 1H), 0.82 (d,> 7.2Hz, 3H).

¹³ C NMR (75 MHz, CDCl ₃ ): delta 170.0, 156.0, 148.4, 147.1, 144.3, 143.0, 138.7, 133.8,

130.5, 129.4, 120.6, 120.4, 117.6, 117.5, 117.0,

113.5, 112.5,112.4, 101.1, 74.1, 66.8, 60.4, 59.3, 56.9,56.6,56.3,55.4,48.7,41.6,40.1,26.2,25.0, 17.6, 15.4, 9.1.

ESI-MS m / z: Calculated for C ₃₅ H _3? F ₃ N ₅ O ₇ :

685.69. Found (M + H) ⁺ : 686.3.

65893 Bl

Example 49

To a solution of 54 (100 mg, 0.145 mmol) in dichloromethane (4 ml) at 23 ° C was added acetic acid (40 ml), (PPh ₃ ) ₂ PdCl ₂ (8.4 mg, 0.012 mmol) and Bu ₃ SnH (15.1 ml, 0.56 mmol). The reaction mixture was stirred for 2 h at this temperature and then poured into a pressure column (silica, gradient hexane to hexane: ethyl acetate 2: 1) to give 55 (90 mg, 96%) as a white solid. .

Rf: 0.6 (hexane: ethyl acetate 1: 2). 1 H NMR (300 MHz, CDCl ₃ ): delta 7.55 (d, J = 7.2 Hz, 1H), 6.45 (s, 1H), 5.90 (d, J = 1.2 Hz, 1H), 5.82 (d,> 1.2 Hz. 1H), 5.37 (t,> 6.0 Hz, 1H), 4.15 (d,> 2.1 Hz,

1H), 4.04 (d,> 1.8 Hz, 1H), 3.70 (s, 3H), 3.663.53 (m, 2H), 3.37-3.21 (m, 2H), 3.19-3.15 (d,> 11.7 Hz, 1H ), 3.08-3.00 (m, 2H), 2.56 (d,> 18.3 Hz, 1H), 2.30 (s, 3H), 2.24 (s, 3H), 2.04 (s, 3H), 1.91 (dd, J = 12.0) Hz, J = 15.6 Hz, 1H), 0.84 (d,> 6.9 Hz, 3H).

¹³ C NMR (75 MHz, CDCl ₃ ): delta 170.1,

156.3, 147.3, 144.9, 144.4, 143.3, 136.7, 130.7,

129.3, 120.6, 117.6, 117.4, 114.4, 112.1, 107.7, 101.0, 85.8, 60.5, 59.3, 56.5, 56.4, 56.2, 55.2,

48.9, 41.6, 40.9, 25.7, 25.3, 18.0, 15.6, 8.7.

ESI-MS m / z: Calculated for C ₃₂ H _3J F ₃ N ₅ O _7:

645.63. Found (M + H) ⁺ : 646.2.

Example 50

To a solution of 17 (200 mg, 0.288 mmol) in dichloromethane (1.44 ml) was added trifluoroacetic acid (888 microl, 11.53 mmol) and the reaction mixture was stirred for 4 h at 23 ° C. The reaction mixture was fixed at 0 ° C with saturated aqueous sodium bicarbonate solution (60 ml) and extracted with ethyl acetate (2x70 ml). The combined organic layers were dried over sodium sulfate and concentrated in vacuo to give 56 (147 mg, 93%) as a white solid, which was used in subsequent reactions without further purification.

Rf: 0.19 (ethyl acetate: methanol 5: 1). 1 H NMR (300 MHz, CH ₃ OD): delta 6.48 (s, 1H), 5.88 (d,> 0.9 Hz, 1H), 5.81 (d,> 0.9 Hz, 1H), 4.35 (d,> 2.4 Hz, 1H ), 4.15 (d,> 1.8 Hz, 1H), 3.99-3.98 (m, 1H), 3.70 (s, 3H), 3.52-2.96 (m, 7H), 2.68 (d, ₅₀ > 18.3 Hz, 1H). 2.24 (s, 3H), 2.23 (s, 3H), 2.06

65893 B1 (s, 3H), 1.85 (dd, 1 = 11.7 Hz, J ₂ = 15.6 Hz, 1H), 0.91 (d, 1 = 6.6 Hz, 3H).

¹ C NMR (75 MHz, CH ₃ OD): delta 173.2,

149.1, 145.6, 144.9, 138.0, 132.2, 130.6, 121.4,

120.6, 119.6, 117.4, 114.3, 109.2, 102.5, 82.3,

60.4,58.4,58.3,57.8,56.6,50.1,42.3,41.6,27.8,

26.2, 19.5, 15.5, 9.8.

ESI-MS m / z: Calculated for C ₂₉ H ₃₅ N ₅ O ₆ : 549.62. Found (M + H) ⁺ : 550.3.

Example 51

C ₆ H _with NCS. CH ₂ CI ₂

1.5h, 23 ° C

OMe

Me -A?

HF

NHCSNHPh

Me

To a solution of 56 (10 mg, 0.018 mmol) in dichloromethane (0.4 ml) was added phenylisothiocyanate (13 microl, 0.109 mmol) and the reaction mixture was stirred for 1.5 h at 23 ° C. The reaction mixture was concentrated in vacuo and the residue purified by flash column chromatography (silica, gradient hexane to hexane: ethyl acetate 1: 1) to give 57 (8 mg, 65%) as a white solid.

Rf: 0.57 (ethyl acetate: methanol 5: 1). 1 H NMR (300 MHz, CDCl ₃ ): delta 7.88 (bs, 1H), 7.41-7.36 (m, 2H), 7.27-7.22 (m, 1H), 7.02-7.00 (d, J = 7.8 Hz, 2H). 6.71 (d, J = 7.2 Hz, 1H), 6.31 (s, 1H), 6.17 (bs, 1H), 5.93 (d, J = 1.2 Hz, 1H), 5.83 (d, J = 1.2)

Hz, 1H), 5.55 (bs, 1H), 5.20-5.17 (m, 1H), 4.16 20 (d, J = 1.8 Hz, 1H), 4.05 (bs, 1H), 4.02 (d, J = 2.4)

Hz, 1H), 3.79 (s, 3H), 3.75-3.71 (m, 1H), 3.35 (d, J = 7.8 Hz, 1H), 3.28-3.19 (m, 2H), 3.12-2.97 (m, 2H) , 2.50 (d, J = 18.3 Hz, 1H), 2.32 (s, 3H), 2.21 (s, 3H), 2.15 (dd, J = 11.4 Hz, J = 15.9 Hz, 1H), 25 1.95 (s, 3H) ), 0.88 (d,> 6.9 Hz, 3H).

¹³ C NMR (75 MHz, CDCl ₃ ): delta 178.5,

171.7, 147.2, 145.0, 144.3, 143.3, 137.0, 135.7,

130.6, 130.4, 129.6, 127.5, 124.3, 120.6, 117.7,

117.2, 115.3, 112.1, 108.3, 100.9, 60.9, 59.5,, ₃₀ 56.7, 56.5, 56.2, 55.2,54.1,41.7,41.1,26.3,25.4,

18.5, 15.8, 9.0.

ESI-MS m / z: Calcd. _n NO, S: up to 4U ο o

684.81. Found (M + H) ⁺ : 685.3.

Example 52

OMe

Me

Me ₀ Y ^ NHCSNAcPh

Me

65893 Bl

Acetyl chloride (4.67 microl, 0.065 mmol) and pyridine (5.3 microl, 0.065 mmol) were added to a solution of 57 (45 mg, 0.065 mmol) in dichloromethane (0.5 ml) at 0 ° C. The reaction mixture was stirred for 3 h and then the solution was diluted with dichloromethane (10 ml) and washed with 0.1N hydrochloric acid (5 ml). The organic layer was dried over sodium sulfate, filtered and the solvent was removed under reduced pressure. The residue was purified by flash chromatography (RP-18, CH ₃ CI: water 40:60) to give 58 (14 mg, 28%) as a white solid.

Rf: 0.34 (CH ₃ CN: water 7:15). NMR (300 MHz, CDCl ₃ ): delta 11.90 (d,> 6.6 Hz, 1H), 7.457.40 (m, 3H), 7.18-7.15 (m, 2H), 6.58 (s, 1H).

6.00 (d,> 1.2 Hz, 1H), 5.89 (d,> 1.2 Hz, 1H),

5.70 (s, 1H), 5.37 (t,> 4.8 Hz, 1H), 4.48 (m, 1H), (bs, 3H), 4.07 (bs, 2H), 3.85-3.75 (m, 1H), 3.70 (s , 3H), 3.46-3.41 (m, 2H), 3.24-3.20 (m, 1H), 5 3.00-2.95 (m, 1H), 2.87-2.75 (m, 1H), 2.31 (s.

3H), 2.28 (s, 3H), 2.24 (s, 3H), 2.00 (s, 3H), 1.85 (dd, J = 11.4 Hz, J = 15.6 Hz, 1H), 1.66 (s, 3H), 0.82 ( d, J = 6.0 Hz, 3H).

¹³ C NMR (75 MHz, CDCl ₃ ): delta 182.6,

174.3, 171.0, 146.6, 144.6, 142.7, 142.3, 140.7,

140.2, 131.3, 129.8, 129.3, 128.9, 128.8, 121.5,

120.4, 117.3, 116.6, 112.8, 112.0, 111.3, 101.5,

60.5, 59.0, 57.6,56.2, 55.9, 55.3, 55.1,41.6, 39.4,

27.8, 26.5, 24.8, 20.2, 17.1, 15.5, 9.3.

ESI-MS m / z: Calculated for C. _n H..NO „S:

768.88. Found (M + H) ⁺ : 769.2.

Me

59

To a solution of 57 (130 mg, 0.189 mmol) in dioxane (1 ml) was added 5.3 N hydrochloric acid / dioxane (1.87 ml) and the reaction mixture was stirred at 23 ° C for 4 h. Dichloromethane (15 ml) and water (10 ml) were then added to the reaction mixture and the organic layer was decanted. The aqueous phase was basified with saturated aqueous sodium bicarbonate solution (60 ml) (pH 8) at 0 ° C and then extracted with ethyl acetate (2 x 50 ml). The combined organic extracts were dried over sodium sulfate and concentrated in vacuo to give 59 (63 mg, 70%) as a white solid.

Rf: 0.15 (ethyl acetate: methanol 5: 1). NMR 30 (300 MHz, CDCl ₃ ): delta 6.67 (s, 1H), 5.99 (d,> 0.9 Hz, 1H), 5.91 (d,> 1.2 Hz, 1H), 5.10 (bs, 1H), 4.32 ( d,> 7.2 Hz, 1H), 4.25 (dd, J = 3.6 Hz, J = 9.3 Hz, 1H), 3.7 (s, 3H), 3.71 (m, 2H), 3.50 (dd, J = 2.4 Hz, J = 15.9 Hz, 1H), 3.42-3.37 (m, 35 2H), 3.16 (dd, J = 3.6 Hz, J = 12.9 Hz, 1H), 2.57 (dd, J = 9.3 Hz, J = 12.9 Hz, 1H) , 2.27 (s, 3H), 2.11 (s, 3H), 1.91 (dd, J = 12.0 Hz, J = 15.9 Hz, 1H).

ESI-MS m / z: Calculated for C ,, HN O _d:

478.5. Found (M + H) ⁺ : 479.3.

Example 54

65893 Bl

To a solution of 43 (20 mg, 0.0338 mmol) in dichloromethane (0.3 ml) at 0 ° C was added sinamoyl chloride (5.63 ml, 0.0338 mmol) and pyridine (2.73 ml, 0.0338 mmol). The reaction mixture was stirred for 1 h and then the solution was diluted with dichloromethane (10 ml) and washed with 0.1 N hydrochloric acid (5 ml). The organic layer was dried over sodium sulfate, filtered and the solvent was removed under reduced pressure. The residue was purified by flash column chromatography (silica, ethyl acetate: methanol 20: 1) to give 60 (22 mg, 90%) as a white solid.

Rf: 0.56 (ethyl acetate: methanol 5: 1). 1 H NMR (300 MHz, CDCl ₃ ): delta 7.51 (s, 1H), 7.50-7.47 (m, 2H), 7.36-7.35 (m, 2H), 6.43 (s, 1H), 6.36 (brd,> 15.9 Hz) , 2H), 6.01 (d,> 1.5 Hz, 1H), 5.90 (brd,> 1.5 Hz, 2H), 5.42 (t, J = 6.0 Hz, 1H), 4.124.07 (m, 3H), 3.96-3.95 (m, 1H), 3.73 (bs, 3H), 3.58 (bs, 2H), 3.39 (d,> 8.7 Hz, 1H), 3.25 (d,> 11.7 Hz, 1H), 3.00 (dd, J = 7.5 Hz) , J = 17.7 Hz, 1H), 2.78 (d,> 15.9 Hz, 1H), 2.67 (d,> 16.5 Hz, 1H), 2.29 (s, 6H), 2.23 (s, 3H), 1.99 (s, 3H) ), 1.82 (dd, 1 = 11.4 Hz, J = 15.6 Hz, 1H), 0.83 (d, J = 6.0 Hz, 3H).

¹³ C NMR (75 MHz, CDCl ₃ ): delta 172.0,

165.0, 146.9, 144.6, 143.1, 141.0, 140.5, 134.8,

131.0, 129.7, 129.1, 128.8, 127.8, 125.5, 123.8,

123.0, 121.1, 120.5, 117.7, 116.9, 112.8, 112.0,

101.9, 60.6, 59.2, 57.1, 56.4, 55.9, 55.3, 48.8,

41.7, 40.0, 26.5, 25.1, 20.3, 18.5, 15.7, 9.3.

ESI-MS m / z: Calculated for C ₄₀ H ₄₃ N ₅ O _g :

721.8. Found (M + H) ⁺ : 722.3.

Example 55

To a solution of 45 (19 mg, 0.0364 mmol) in dichloromethane (0.3 ml) at 0 ° C was added heptafluorobutyryl chloride (5.44 microl, 0.0364 mmol) and pyridine (2.95 microl, 0.0364 mmol). The reaction mixture was stirred for 1 h and then the solution was diluted with dichloromethane (10 ml) and washed with 0.1 N hydrochloric acid (5 ml). The organic layer was dried over sodium sulfate, filtered and the solvent was removed under reduced pressure. The residue was purified by flash column chromatography (silica, ethyl acetate: methanol 20: 1) to give 61 (11.7 mg, 45%) as a white solid.

Rf: 0.76 (ethyl acetate: methanol 5: 1).

NMR (300 MHz, CDCl ₃ ): delta 6.46 (s,

W), 6.12 (bs, 1H), 5.98 (d,> 1.2 Hz, 1H), 5.93 (d,> 1.2 Hz, 1H), 5.72 (bs, 1H), 4.13-4.11 (m, 2H), 4.0 ( d,> 2.4 Hz, 1H), 3.98-3.96 (m, 1H), 3.73 (s, 3H), 3.39 (d,> 7.5 Hz, 1H), 3.39-3.28 (m, 2H), 3.09 (dd, J = 8.1 Hz, J = 18.0 Hz, 1H),

2.80 (d,> 16.2 Hz, 1H), 2.46 (d,> 18.3 Hz, 1H), 2.32 (s, 6H), 2.21 (s, 3H), 1.99 (s, 3H), 1.80 (dd, J = 12.0 Hz, J = 16.2Hz, 1H).

ESI-MS m / z: Calculated for C ₃₂ H ₃₁ F ₇ N ₄ O _? :

716.6. Found (M + H) ⁺ : 717.2.

65893 Bl

To a solution of 43 (24 mg, 0.04 mmol) in dichloromethane (0.3 ml) at 0 ° C was added butyryl chloride (4.15 microl, 0.04 mmol) and pyridine (3.28 microl, 0.04 mmol). The reaction mixture was stirred for 1 h and then the solution was diluted with dichloromethane (10 ml) and washed with 0.1 N hydrochloric acid (5 ml). The organic layer was dried over sodium sulfate, filtered and the solvent was removed under reduced pressure. The residue was purified by flash column chromatography (silica, ethyl acetate: methanol 20: 1) to give 62 (24 mg, 90%) as a white solid.

Rf: 0.35 (ethyl acetate: methanol 5: 1). 1 H NMR (300 MHz, CDCl ₃ ): delta 6.47 (s, 1H), 6.10 (d,> 6.5 Hz, 1H), 6.0 (d,> 1.5 Hz, 1H), 5.91 (d,> 1.5 Hz, 1H) , 5.86 (bs, 1H), 5.31 (d,> 6.9 Hz, 5H), 4.11-4.06 (m, 3H), 3.85-3.81 (m, 1H), 3.75 (s, 3H), 3.59-3.53 (m , 2H), 3.38 (d,> 7.5 Hz, 1H), 3.27 (m, 2H), 3.0 (dd, J = 7.8Hz, J = 17.4Hz, 1H), 2.79 (d,> 15.3 Hz, 1H). 2.63 (d,> 17.7 Hz, 1H), 2.31 (s, 3H), 2.0 (s, 3H), 1.80 (dd, J = 12.0 Hz, 10 J = 15.9 Hz, 1H), 1.58 (q,> 7.2 Hz , 2H), 0.89 (t,> 7.2 Hz, 3H), 0.76 (d,> 6.6 Hz, 3H).

ESI-MS m / z: Calculated for C ₃₅ H ₄₃ N _s O _g :

661.64. Found (M + H) ⁺ : 662.3.

Example 57

C ₉ H ₇ SO, Ru, CH ₂ Cl ₂ .0 °, 1h.

To a solution of 43 (19 mg, 0.0364 mmol) in dichloromethane (0.3 ml) at 0 ° C was added sinamoyl chloride (6.06 ml, 0.0364 mmol) and pyridine (2.95 microl, 0.0364 mmol). The reaction mixture was stirred for 1 h and then the solution was diluted with dichloromethane (10 ml) and washed with 0.1 N hydrochloric acid (5 ml). The organic layer was dried over sodium sulfate, filtered and the solvent was removed under reduced pressure. The residue was purified by flash column chromatography (silica, ethyl acetate: methanol 20: 1) to give 63 (20.1 mg, 85%) as a white solid.

Rf: 0.65 (ethyl acetate: methanol 5: 1). 1 H NMR (300 MHz, CDCl ₃ ): delta 7.39-7.29 (m, 5H), 6.42 (s, 1H), 6.01 (d,> 1.5 Hz, 1H), 5.92 (d,> 1.5 Hz, 30 IH). 5.73 (bs, 1H), 5.24 (t,> 6.8 Hz, 1H), 4.124.08 (m, 3H), 3.66-3.64 (m, 2H), 3.58 (bs, 3H), 3.36 (d,> 8.7 Hz , 1H), 3.29 (d,> 12.0 Hz, 1H), 2.98 (dd, J = 8.1 Hz, J = 18.0Hz. 1H), 2.33 (s, 6H), 2.29 (s, 3H), 2.01 (s. 3H), 1.84 (dd, J = 12.0 Hz, 35 J ₂ = 15.9Hz, 1H).

ESI-MS m / z: Calculated for C ₃₇ H _3g N ₄ O _7: 650.72. Found (M + H) ⁺ : 651.2.

Example 58

65893 Bl

To a solution of 43 (20 mg, 0.0338 mmol) in dichloromethane (0.3 ml) at 0 ° C was added 3chloropropionyl chloride (3.22 microl, 0.0338 mmol) and pyridine (2.73 microl, 0.0338 mmol). The reaction mixture was stirred for 1 h and then the solution was diluted with dichloromethane (10 ml) and washed with 0.1 N hydrochloric acid (5 ml). The organic layer was dried over sodium sulfate, filtered and the solvent was removed under reduced pressure. The residue was purified by flash column chromatography (silica, ethyl acetate: methanol

20: 1) to give 64 (20.5 mg, 89%) as a white solid.

Rf: 0.32 (ethyl acetate: hexane 5: 1). 1 H NMR (300 MHz, CDCl 3): delta 6.48 (s, 3H), 6.28 (m, 5H), 5.99 (d,> 1.2 Hz, 1H), 5.91 (d,> 1.2 Hz, 1H), 5.86 (bs , 1H), 5.31 (m, 1H), 4.08-4.07 (m, 3H), 3.75 (s, 3H), 3.72-3.53 (m, 5H), 3.39 (d,> 8.1 Hz, 1H), 3.24 (d ,> 12.0Hz, 1H), 3.00 (dd, J = 8.1 Hz, J = 18.0 Hz, 1H), 2.79 (d,> 13.5 Hz, 1H), 2.50 10 (t,> 6.3 Hz, 2H), 2.32 ( s, 3H), 2.28 (s, 3H), 2.25 (s, 3H), 2.0 (s, 3H), 1.79 (dd, J = 12.3 Hz, J = 14.8 Hz, 1H), 0.81 (d, J = 6.3 Hz, 3H).

Example 59

To a solution of 43 (19 mg, 0.0364 mmol) in dichloromethane (0.3 ml) at 0 ° C was added butyryl chloride (3.78 microl, 0.0364 mmol) and pyridine (2.95 microl, 0.0364 mmol). The reaction mixture was stirred for 1 h and then the solution was diluted with dichloromethane (10 ml) and washed with 0.1N hydrochloric acid (5 ml). The organic layer was dried over sodium sulfate, filtered and the solvent was removed under reduced pressure. The residue was purified by flash column chromatography (silica, ethyl acetate: methanol 20: 1) to give 64 (19 mg, 87%) as a white solid.

Rf: 0.32 (ethyl acetate: methanol 5: 1). 1 H NMR (300 MHz, CDCl ₃ ): delta 6.50 (s, 1H), 5.98 (d,> 1.5 Hz, 1H), 5.91 (d,> 1.5 Hz, 1H), 5.75 (s, 1H), 5.01 (t ,> 6.4 Hz, 1H), 4.10-4.09 (m, 1H), 4.06 (d,> 2.1 Hz, 1H), 4.03-4.02 (m, 1H), 3.76 ₃₀ (s, 3H), 3.67-3.60 (m , 1H), 3.42-3.35 (m, 2H), 3.29 (d,> 12.0 Hz, 1H), 3.02 (dd, J = 7.8 Hz,

J = 17.7 Hz, 1H), 2.79 (d,> 14.1 Hz, 1H), 2.56 (d,> 18.3 Hz, 1H), 2.32 (s, 3H), 2.31 (s, 3H), 2.25 (s, 3H) , 1.78 (dd, J = 12.0 Hz, J = 15.9 Hz, 1H), ₃₅ 1.63 (s, 3H), 1.53-1.46 (m, 2H), 1.28-1.16 (m, 2H), 0.68 (t,> 7.2 Hz, 3H).

ESI-MS m / z: Calcd. For C ₃₂ H ₃ N ₄ O ₇ : 590.67. Found (M + H) ⁺ : 591.2.

Example 60

65893 Bl

To a solution of 50 (31.7 mg, 0.044 mmol) in CH 2 CN / water (1.5 ml / 0.5 ml) was added silver nitrate (225 mg, 1.32 mmol) and the reaction mixture was stirred at 23 ° C for 17 h. Then, brine (10 ml) and aqueous saturated sodium bicarbonate solution (10 ml) were added to the reaction mixture at 0 ° C and the reaction mixture was stirred for 15 min, filtered through a pad of celite and washed with dichloromethane ( 20 ml). The solution is decanted white solid.

Rf: 0.26 (ethyl acetate: methanol 5: 1). 1 H NMR (300 MHz, CDCl ₃ ): delta 7.66-7.42 (m, 4H), 7.20 (bs, 1H), 6.44 (s, 1H), 5.97 (b,> 1.2 Hz, 1H), 5 5.90 (d. > 1.2 Hz, 1H), 5.76 (bs, 1H), 5.28 (bs, 1H), 4.54 (bs, 1H), 4.43 (bs, 1H), 4.00 (bs, 1H), 3.68-3.57 (m, 4H) , 3.47 (d,> 3.3 Hz, 1H), 3.40 (d,> 11.7 Hz, 1H), 3.17 (d,> 6.9 Hz, 1H), 2.92 (dd, J = 8.1 Hz, J = 17.7 Hz, 1H) , 2.74 (d,> 17.1 10 Hz, 1H), 2.48 (d,> 18.6 Hz, 1H), 2.32 (s, 6H) and the organic layer was dried over sodium sulfate and concentrated in vacuo. The residue was purified by flash column chromatography (silica, ethyl acetate: MeOH).

2.28 (s, 3H), 1.99 (s, 3H), 1.76 (dd, J = 12.0 Hz, J = 16.2Hz, 1H).

ESI-MS m / z: Calculated for C ₃₇ H _3g F ₃ N ₃ O _8: 709. Found (M ⁺ -17): 692.3.

ethanol 5: 1) to give 66 (16 mg, 51%) 15

Example 61

To a solution of 53 (57 mg, 0.0828 mmol) in CH ₃ CN / water (1.5 ml / 0.5 ml) was added silver nitrate (650 mg, 3.81 mmol) and the reaction mixture was stirred at 23 ° C for 24 h. Then, brine (10 ml) and aqueous saturated sodium bicarbonate solution (10 ml) were added to the reaction mixture at 0 ° C and the reaction mixture was stirred for 15 min, filtered through a pad of celite and washed with dichloromethane ( 20 ml). The solution was decanted and the organic layer was dried over sodium sulfate and concentrated in vacuo. The residue was purified by flash column chromatography (silica, ethyl acetate: methanol 5: 1) to give 67 (28 mg, 50%) as a white solid.

Rf: 0.28 (ethyl acetate: methanol 10: 1). 1 H NMR (300 MHz, CDCl ₃ ): delta 6.47 (s, 1H), 5.97 (s, 1H), 5.35 (bs, 1H), 4.51 (bs, 1H), 4.41 (bs, 1H), 4.12-4.05 ( m, 1H), 4.00 (d,> 2.7 Hz, 1H), 3.77 (s, 3H), 3.64 (bs, 1H), 3.46 (d,> 3.3 Hz, 1H), 3.34 (d,> 11.4 Hz, 1H ), 3.18 (d,> 7.5 Hz, 1H), 2.95 (dd, J = 8.4 Hz, J = 18.3 Hz, 1H), 2.70 (d, ₃₅ > 15.6 Hz, 1H), 2.48 (d,> 17.7 Hz, 1H), 2.28 (s, 3H), 2.27 (s, 3H), 2.26 (s, 3H), 1.98 (s, 3H), 1.68 (dd, J = 12.0 Hz, J = 15.6 Hz, 1H). 0.86 (d, > 6.3 Hz, 3H).

ESI-MS m / z: Calculated for C _J2 H ₃₇ F ₃ N ₄ O _9: 678.66. Found (M ⁺ -17): 661.2.

Example 62

65893 Bl

To a solution of 48 (32 mg, 0.0529 mmol) in CH 2 CN / water (1.5 ml / 0.5 ml) was added silver nitrate (270 mg, 1.58 mmol) and the reaction mixture was stirred at 23 ° C for 24 h. Then, brine (10 ml) and aqueous saturated sodium bicarbonate solution (10 ml) were added to the reaction mixture at 0 ° C and the reaction mixture was stirred for 15 min, filtered through a pad of celite and washed with dichloromethane ( 20 ml). The solution was decanted and the organic layer was dried over sodium sulfate and concentrated in vacuo. The residue was purified by flash column chromatography (silica, ethyl acetate: methanol 5: 1) to give 68 (18 mg, 56%) as a white solid.

Rf: 0.40 (ethyl acetate: methanol 5: 1). 1 H NMR (300 MHz, CDCl ₃ ): delta 6.50 (s, 1H), 5.95 (d, 5> 1.2 Hz, 1H), 5.88 (d,> 1.2 Hz, 1H), 5.23 (d,> 6.9 Hz, 1H ), 4.45 (d,> 3.3 Hz, 1H), 4.38 (s, 1H), 4.01 (d,> 2.4 Hz, 1H), 3.78 (m, 3H), 3.413.37 (m, 1H), 3.17-3.15 (m, 1H), 2.96 (dd, J = 7.8 Hz, J = 18.0 Hz, 1H), 2.70 (d,> 15.3 Hz, 1H), 10 2.40 (d,> 18.0 Hz, 1H), 2.30 (s. 6H), 2.27 (s, 3H), 1.76-1.65 (m, 1H), 1.35-1.25 (m, 2H), 0.890.82 (m, 1H), 0.69 (d,> 6.6 Hz, 3H), 0.58 ( d, J = 6.6 Hz, 3H).

Example 63

NH I

NH I cM69

To a solution of 51 (27 mg, 0.04 mmol) in CH ₃ CN / water (1.5 ml / 0.5 ml) was added silver nitrate (204 mg, 1.19 mmol) and the reaction mixture was stirred at 23 ° C for 24 h. Then, brine (10 ml) and aqueous saturated sodium bicarbonate solution (10 ml) were added to the reaction mixture at 0 ° C and the reaction mixture was stirred for 15 min, filtered through a pad of celite and washed with dichloromethane ( 20 ml). The solution was decanted and the organic layer was dried over sodium sulfate and concentrated in vacuo. The residue was purified by flash column chromatography (silica, ethyl acetate: methanol 5: 1) to give 69 (10 mg, 38%) as a white solid.

Rf: 0.38 (ethyl acetate: methanol 5: 1). 1 H NMR (300 MHz, CDCl ₃ ): delta 6.48 (s, 1H), 6.16 (bs, 30 IH), 5.98 (d,> 1.5 Hz, 1H), 5.89 (d,> 1.5 Hz, 1H), 5.33 ( t,> 6.0 Hz, 1H), 4.50 (m, 1H), 4.40 (m, 1H), 4.11-4.09 (m, 1H), 4.00 (d,> 2.6 Hz, 1H), 3.78 (s, 3H). 3.41-3.32 (m, 3H), 3.18 (d,> 8.4 Hz, 1H), 2.94 (dd, J = 8.4 Hz, J = 18.3 Hz, 1H), 35 2.70 (d,> 14.4 Hz, 1H), 2.45 (d,> 18.3 Hz, 1H), 2.31 (s, 3H), 2.28 (s, 3H), 2.27 (s, 3H), 2.04 (s, 3H), 2.01-1.86 (m, 3H), 1.73 (m , 1H), 0.87 (d,> 6.3 Hz, 6H).

Example 64

65893 Bl

To a solution of 63 (15 mg, 0.023 mmol) in CH 2 CN / water (1.5 ml / 0.5 ml) was added silver nitrate (118 mg, 0.691 mmol) and the reaction mixture was stirred at 23 ° C for 24 h. Then, brine (10 ml) and aqueous saturated sodium bicarbonate solution (10 ml) were added to the reaction mixture at 0 ° C and the reaction mixture was stirred for 15 min, filtered through a pad of celite and washed with dichloromethane ( 20 ml). The solution was decanted and the organic layer was dried over sodium sulfate and concentrated in vacuo. The residue was purified by flash column chromatography (silica, ethyl acetate: methanol 5: 1) to give 70 (20.1 mg, 85%) as a white solid.

Rf: 0.43 (ethyl acetate: methanol 5: 1). 1 H NMR (300 MHz, CDCl ₃ ): delta 7.38-7.28 (m, 5H), 6.48 (s, 1H), 5.98 (d,> 1.5 Hz, 1H), 5.91 (d, J = t.5 Hz, 1H ), 5.75 (bs, 1H), 5.38 (brd, 1H), 5.30 (bs, 1H), 4.53 (m, 1H), 4.42 (m, 1H), 4.02 (d,> 2.7 Hz, 1H), 3.78- 3.65 (m, 5H), 3.46-3.40 (m, 2H), 3.17 (d,> 7.8 Hz, 1H), 2.94 (dd, J = 7.8 Hz, J = 17.7 Hz, 1H), 2.73 (d,> 16.8 Hz, 1H), 2.45 (d,> 18.0

Hz, 1H), 2.31 (s, 3H), 2.28 (s, 3H), 1.97 (s, 3H), 1.77 (dd, J = 12.0 Hz, J = 15.3 Hz, 1H).

Example 65

To a solution of 65 (25 mg, 0.042 mmol) in CH ₃ CN / water (1.5 ml / 0.5 ml) was added silver nitrate (215.56 mg, 1.269 mmol) and the reaction mixture was stirred at 23 ° C for 24 h. Then, brine (10 ml) and aqueous saturated sodium bicarbonate solution (10 ml) were added to the reaction mixture at 0 ° C and the reaction mixture was stirred for 15 min, filtered through a pad of celite and washed with dichloromethane ( 20 ml). The solution was decanted and the organic layer was dried over sodium sulfate and concentrated in vacuo. The residue was purified by flash column chromatography (silica, ethyl acetate: methanol 5: 1) to give 71 (16 mg, 65%) as a white solid.

Rf: 0.05 (ethyl acetate: methanol 5: 2). 1 H NMR (300 MHz, CDCl ₃ ): delta 6.50 (s, 1H), 5.95 (d,> 1.5 Hz, 1H), 5.78 (s, 1H), 5.19 (bs, 1H), 4.45 (d,> 3.3 Hz) , 1H), 4.37 (bs, 1H), 4.11 (brd,> 4.8 Hz, 1H), 4.01 (d,> 2.1 Hz, 1H), 3.76 (s, 1H), 3.713.69 (m, 1H), 3.49 -3.35 (m, 1H), 3.24 (d,> 13.5 Hz, 1H), 3.15 (d,> 9.3 Hz, 1H), 2.95 (dd, J = 8.1)

Hz, J = 17.7Hz, 1H), 2.70 (d,> 15.6Hz, 1H), 2.40 (d,> 18.0 Hz, 1H), 2.31 (s, 3H), 2.29 (s, 3H), 2.28 (s. 3H), 1.96 (s, 3H), 1.75-1.66 (m, 3H), 1.521.17 (m, 2H), 0.66 (t,> 7.2 Hz, 3H).

Fermentation methods

Example A

YMP3 embryo medium containing 1% glucose, 0.25% bovine extract, 0.5% bactopeptone, 0.25% sodium chloride, 0.8% calcium carbonate was inoculated with 0.1% frozen vegetative mass of microorganisms from Pseudomonas fluorescence strain A22, and scscores and fluorescent in a rotary shaker (250 o6./min) at 27 ° C. After incubation for 30 hours, the culture was added with stirring to a fermentation vessel containing medium consisting of 2% dextrose, 4% mannitol, 2% dried beer yeast (Vitalevor® Biolux, Belgium), 1% ammonium sulfate, 0.04% acid potassium phosphate, 0.8% potassium chloride, 0.001% ferric chloride, 0.1% L-Tyr, 0.8% calcium carbonate, 0.05% polypropylene glycol PPG-2000,0.2% silicone (ASSAF-100, RHODIA UK). Sterilization was carried out at 122 ° C

65893 Bl for 30 min. The inoculated volume is 2% (v / v). The temperature is 27 ° C (0 to 16 h) and 24 ° C 16 to the end of the process (41 h). Dilution oxygen pressure is above 25%. The pH was adjusted to 6.0 using dilute sulfuric acid from the 28th hour until the end of the process. The overpressure is 0.5 bar. From the 16th hour to the end of the process (running for 2 days), 1% mannitol or sorbitol was added and 2% for the three-day fermentation process.

After 41 or 64 hours, the fermentation broth should be extracted to restore safracin B or the clarified broth treated with potassium cyanide to restore safracin B-cyano.

Example B

Preparation of safraciv B-cyano from crude extract

Solids are separated by clarification or filtration of the fermentation broth at pH 6. The clarified broth was adjusted to pH 9.5 using a dilute sodium hydroxide solution and extracted twice with 2: 1 (v / v) ethyl acetate, methylene chloride or butyl acetate. The extraction was carried out in a stirring vessel for 20 min while maintaining the temperature of the mixture from 8 to 10 ° C. The two phases are separated by liquid-liquid centrifugation. The organic phase was dried over anhydrous sodium sulfate or by freezing and then filtered to remove the ice. The organic phase (ethyl acetate layer) was evaporated to give a crude oil extract,

Example B

Preparation of safracin B-cyano from clear broth

Solids are separated by clarification or filtration of the fermentation broth at pH 6. The clarified broth was adjusted to pH 3.9 using concentrated acetic acid. 0.5 g / Ι potassium cyanide was added to the clarified broth and incubated at 20 ° C for 1 hour with stirring. The temperature was then lowered to 15 ° C and the pH adjusted to 9.5 using a dilute sodium hydroxide solution and extracted with 2: 1.5 (v / v) ethyl acetate. The extraction was carried out in a stirring vessel for 20 min while maintaining the temperature of the mixture from 8 to 10 ° C. The two phases are separated by liquid-liquid centrifugation. The organic phase was dried over anhydrous sodium sulfate. The organic phase (ethyl acetate layer) was evaporated until a crude oil extract was obtained. This extract was purified by flash chromatography (silica, gradient 20: 1 to 10: 1 to 5: 1 ethyl acetate: methanol) to quantitatively yield compound 2 as a light yellow solid,

Rf: 0.55 (ethyl acetate-methanol 5: 1): 19.9 min [HPLC, Delta Pack C4.5 microm, 300 A, 150 x 3 mm, lambda = 215 nm, flow = 0.7 ml / min, temperature = 50 ° C. gradient: aqueous CH ₃ CN NaOAc (10 mM) 85-70% (20 ')].

NMR (300 MHz, CDCl ₃ ): delta 6.54 (dd, J = 4.4 Hz, J = 8.4 Hz, 1H), 6.44 (s, 1H), 4.12 (d, J = 2.4 Hz, 1H), 4.04 (d , J = 2.4 Hz, 1H), 4.00 (s, 3H), 3.87 (bs, 3H), 3.65 (ddd, J = 1.5 Hz, J = 8.7 Hz, J = 9.9Hz, 1H), 3.35 (brd, J = 8.4Hz, 1H), 3,152.96 (m, 4H), 2.92 (q, J = 7.2 Hz, 1H), 2.47 (d, 1 = 18.3 Hz, 1H), 2.29 (s, 3H), 2.18 (s , 3H), 1.83 (s, 3H), 1.64 (ddd, J = 2.7Hz, J = 11.1 Hz, J = 14.1 Hz, 1H), 0.79 (d, J = 7.2 Hz, 3H).

¹³ CNMR (75 MHz, CDC1 ₃ ): delta 186.0 (q), 175.9 (q), 156.2 (q), 146.8 (q), 142.8 (q), 140.7 (q), 136.6 (q), 130.5 (q) , 128.8 (q), 127.0 (q), 120.5 (s), 117.4 (q), 116.5 (q), 60.8 (t), 60.4 (s), 58.7 (t), 56.2 (s), 55.7 (s) , 54.8 (s), 54.8 (s), 54.4 (s), 50.0 (s), 41.6 (t), 39.8 (d), 25.2 (d), 24.4 (d), 21.2 (t), 15.5 (t) , 8.4 (t).

ESI-MS m / z: Calculated for C ₂₉ H ₃₅ N ₅ O ₆ : 549.6. Found (M + Na) ⁺ : 572.3.

Example D

Medium (501) consisting of 2% dextrose, 4% mannitol, 2% dry brewer's yeast (Vitalevor® Biolux, Belgium), 1% ammonium sulfate, 0.04% acid potassium phosphate, 0.8% potassium chloride, 0.001% ferric chloride 6 -hydrate, 0.1% L-tyrosine, 0.8% calcium carbonate, 0.05% polypropylene glycol PPG-2000 and 0.2% silicone (ASSAF-100, RHODIA UK), poured into a fermentation vessel with a total volume of 75 l and after sterilization, was inoculated with culture (2%) of A2-2 strain (FERM BP-14) and the aerated cultivation under stirring was carried out at from 27 ° C to 24 ° C for 64 h (aeration at 75 Ι / min and stirring at from 350 to 500 rpm). The pH was controlled by automatic feeding with dilute sulfuric acid from the 27th hour until the end of the process. From the 16th hour to the end of the process, 2% is added

65893 Bl mannitol. The culture medium thus obtained (45 L), after separation of the cells by centrifugation, was adjusted to pH 9.5 using a dilute sodium hydroxide solution and extracted twice with 251 ethyl acetate. The mixture was placed in a stirring vessel at 8 ° C for 20 min. The two phases are separated by liquid-liquid centrifugation. The organic phase was frozen at -20 ° C and filtered to remove ice and evaporated to give 40 g of a dark crude oil extract. After introduction of the cyanide group and purification, 3 g of safracinB-cyano are obtained.

Example E

Medium (501) consisting of 2% dextrose, 4% mannitol, 2% dried brewer's yeast (Vitalevor® Biolux, Belgium), 1% ammonium sulfate, 0.02% acid potassium phosphate, 0.2% potassium chloride, 0.001% ferric chloride 6 -hydrate, 0.1% L-tyrosine, 0.8% calcium carbonate, 0.05% polypropylene glycol PPG-2000 and 0.2% silicone (ASSAF-100, RHODIA UK), poured into a fermentation vessel with a total volume of 75 l and, after sterilization, was inoculated with culture (2%) of A2-2 strain (FERM BP-14) and the aerated stirring cultivation was carried out at from 27 ° C to 24 ° C for 41 h (aeration at 75 Ι / min and stirring at from 350 to 500 rpm), the pH is controlled by auto mother liquor with dilute sulfuric acid from the 28th hour until the end of the process. From the 16th hour until the end of the process, 1% mannitol was added. The culture medium thus obtained (45 L), after separation of the cells by centrifugation, was adjusted to pH 3.9 with 200 ml of concentrated acetic acid. 25 g of potassium cyanide 97% were added and, after stirring for 1 h at 20 ° C, the pH was adjusted to 9.5 with the help of 1500 ml of 10% sodium hydroxide solution. It is then extracted with 3 51 ethyl acetate.

The mixture was placed in a stirring vessel at 8 ° C for 20 min. The two phases are separated by liquid-liquid centrifugation. The organic phase was dried over anhydrous sodium sulfate and evaporated to give 60 g of 10 dark oily crude extract.

Chromatography afforded 4.9 g of safracin B-cyano.

Claims (35)

Claims A process for the preparation of a compound of formula (XVIIb): characterized in that: R ¹ is an aminomethylene group, a protected aminomethylene group, a derivative of an amino30 methylene group, a hydroxymethylene group, a protected hydroxymethylene group or a derivative of a hydroxymethylene group; and R ⁴ is -H; or R ¹ and R ⁴ together form a group of formula 35 (IV), (V), (VI) or (VII): 65893 Bl R ⁵ is -H, -OH; R ⁷ is -OCH 3 and R ⁸ is -OH or R ⁷ and R ⁸ together form a -O-CH 2 -O- group; R ¹² is -H, -CH ₃ or -CH ₂ CH ₃ ; RMa _and R14b _and e _{Watts are} _H OR the ONLY β -H, in the other is -OH, -OCH ₃ or -OCH ₂ CH ₃ , or R ^14a and R ^14b together form a ketone group; R ¹⁵ is -H, -OH; R ¹⁸ is -H or -OH; R ²¹ is -H, -OH or -CN; and derivatives; of the 21-cyano compound of formula (XVIb): wherein: R ¹ is an amidomethylene group or an acyloxymethylene group; R ⁵ and R ^{8 are} independently selected from -H, -OH or -OCOCH ₂ OH, or R ⁵ and R ⁸ are both keto and ring A is a p-benzoquinone ring; R ¹² is -H, -CH ₃ or -CH ₂ CH ₃ ; R ^14a and R ^14b are both -H or one is -H and the other is -OH, -OCH ₃ or -OCH ₂ CH ₃ , or R ^14a and R ^14b together form a ketone group; and R ¹⁵ and R ^{18 are} independently selected from -H or -OH, or R ¹⁵ or R ¹⁸ are both keto and ring E is a p-benzoquinone ring; provided that at least one of rings A or E is a p-benzoquinone ring; the reactions according to the method include: a) conversion of the quinone system from ring E into a phenolic system; b) converting the quinone system from ring A into a phenolic system; c) conversion of the phenol system from ring A into a methylenedioxyphenol ring; d) forming a bridged spiro ring system of formula (IV), (V), (VI) or (VII) through positions 1 and 4 in ring B; e) obtaining derivatives; f) converting a compound in which R ¹² is -CH ₃ or into a group in which R ¹² is selected from -H and -CH ₂ CH ₃ ; and g) converting the 21-cyano substituent to a substituent -OH or -H, as appropriate; to obtain the desired compound of formula (XVIIb). A method according to claim 1, characterized in that step a) for converting the quinone system from ring E into a phenolic system is carried out by reduction using hydrogen with a palladium-on-carbon catalyst. The method according to claim 1 or 2, characterized in that step b) for converting the quinone system from ring A into a phenolic system is carried out by reduction using hydrogen with a palladium-on-carbon catalyst. A method according to claim 1, characterized in that it comprises at least steps b), c), e) and g). A method according to any one of claims 1 to 4, characterized in that R ¹ and R ⁴ together form a group of formula (IV), (V), (VI) or (VII), and step d) for forming a bridged spiro ring system of the formula through positions 1 and 4 in ring B is carried out by substitution at position 1 with a bridge-forming reagent to form exendo quinonmetide at position 4, and reaction of the methide with the 1-substituent to obtain spiral ring bridge system. A method according to claim 5, characterized in that the bridge-forming reagent is of formula (XIX) in which Fu is a protected functional group, Prot ³ is a protecting group, and the intersection shows a possible double bond. A method according to claim 6, wherein Fu is a group -NHProt ⁴³ or OProt ⁴ ", where Prot ⁴ " is an amino protecting group and Prot ⁴¹¹ is a hydroxy protecting group. A method according to claim 7, characterized in that the bridge-forming reagent is Int-29 or Int-37: 65893 Bl A method according to any one of claims 5 to 8, characterized in that the formation of the methide involves introducing a hydroxyl group at position 10 at the bond between rings A and B to form a separate structure of formula (XX): in which the group R 'is selected to form the desired group of formula (IV), (V), (VI) or (VII). A method according to claim 9, characterized in that the introduction of a hydroxyl group at position 10 at the bond between rings A and 6 results in a separate structure of formula (XXI): in claim 9. A method according to claim 9 or 10, characterized in that the group R 'is -CHFuCH ₂ -SProt ³ , wherein Fu is a protected functional group and Prot ³ is a thiol-protecting group. A method according to any one of claims 1 to 11, characterized in that step e) for the preparation of derivatives involves acylation. A method according to any one of claims 1 to 12, characterized in that 21 cyano compounds of formula (XVIb) are prepared by introducing a 21-cyano group into saframycin A, saframycin B, saframycin C, saframycin G, saframycin H , saframycin S, 5 saframycin Y ₃ , saframycin Yd _p saframycin Ad ,, saframycin Yd ₂ , saframycin AH ₂ , saframycin AH ₂ Ac, saframycin AH _p saframycin AH, Ac, saframycin AR ₃ , renieramycin A, renieramycin B, renieramycin C renieramycin D, renieramycin E, 10 renieramycin F, xestomycin, saframycin D, saframycin F, saframycin Mx-1, saframycin Mx2, safracin A, or safracin B, or saframycin R. A method according to claim 13, characterized in that the 21-cyan compound 15 of formula (XVIb) is cyanosafracin B, compound (2) of formula: compound 2 A method according to any one of claims 1 to 14, characterized in that R ⁵ in the compound of formula (XVIIb) is alkanoyloxy of 1 to 5 carbon atoms. A method according to any one of claims 1 to 15, characterized in that R ⁵ in the compound of formula (XVIIb) is acetyloxy. A method according to any one of claims 1 to 16, wherein R ¹² is -CH ₃ . A process according to any one of claims 1 to 17, characterized in that R ^14a and R ^14b in the compound of formula (XVIIb) are hydrogen. A method according to any one of claims 1 to 18, characterized in that R ¹⁵ in the compound of formula (XVIIb) is hydrogen. A method according to any one of claims 1 to 19, characterized in that R ¹⁸ in the compound of formula (XVIIb) is hydroxy. 65893 Bl A method according to any one of claims 1 to 20, characterized in that R ²¹ in the compound of formula (XVIIb) is -OH or -CN. A method according to any one of claims 1 to 21, characterized in that R ⁷ and R ⁸ in the compound of formula (XVIIb) together form a group -O-CH ₂ -O-. A method according to any one of claims 1 to 22, characterized in that R ¹ and 5 R ⁴ in the compound of formula (XVIIb) together form a group of formula (IV), (V), (VI) or (VU): A method according to any one of claims 1 to 4 and 12 to 22, characterized in that R ¹ in the compound of formula (XVIIb) is a possibly protected or modified aminomethylene group, possibly protected or modified hydroxymethylene group; and R ⁴ is -H. The method of claim 24, wherein R ¹ in the compound of formula (XVIIb) is a group -CH ₂ -NII, or -CH ₂ -NH-aa, 2 5 where aa is an acylamino acid group. 26. The method of claim 25, wherein the compound of formula (XVIIb) is an N-acyl derivative of the group R ¹ when -CH ₂ NH ₂ or -CH ₂ -NH-aa. ₃ θ The method of claim 26, wherein R ¹ in the compound of formula (XVIIb) is an N-acyl derivative in which the acyl group is of formula -CO-R ^a , wherein R ^a is alkyl, alkoxy, alkenyl , aralkyl, aralkenyl, amino acid acyl or heterocyclyl; each optionally substituted with halo, cyano, nitro, carboxyalkyl, alkoxy, aryl, aryloxy, heterocyclyl, heterocyclyloxy, alkyl, amino or substituted amino; or the acyl group is aa. A method according to claim 25,26 or 40 27, characterized in that more or more aa groups are present and are alanyl, arginyl, aspartyl, asparagyl, cystyl, glutamyl, glutaminyl, glycyl, histidyl, hydroxyprolyl, isoleucyl, leucyl, lysyl, methionyl, phenylalanil, prolyl, seryl, threonyl, tyronyl, tryptophil, tyrosyl or valyl. A method according to any one of claims 1 to 28, characterized in that in each reaction, one or more substituents are protected by a protecting group. 30. A process for the preparation of a compound of formula (XXIIb): characterized in that: R ¹ is -CH ₂ NH ₂ or -CH ₂ H or a protected or modified version of this group and R ⁴ is -H; or R ¹ and R ⁴ together form a group of formula (IV), (V), (VI) or (VII): 65893 Bl R ⁵ is -OH or a protected or modified version of this group; R ¹² is -H, -CH or -CH ₂ CH ₃ ; R ^14a and R ^14b are both -H or one is -H and the other is -OH or a protected or modified version of this group, -OCH ₃ or -OCH ₂ CH ₃ , or Ri4a _and Ri4b _together form a ketone group; R ¹⁵ is -H, -OH or a protected or modified version of this group; R ¹⁸ is -OH or a protected or modified version of this group; and R ²¹ is -H, -OH or -CN; of the 21-cyano compound of formula (XVIb): wherein: R ¹ is an amidomethylene group or an acyloxymethylene group; R ⁵ and R ⁸ , independently of one another, are selected from -H, -OH or -OCOCON, or R ⁵ and R ⁸ are both keto and ring A is a p-benzoquinone ring; R ¹² is -H, -CH ₃ or -CH ₂ CH ₃ ; R14a _and R14b _and e _{Watts are or are diene} T is -H and the other is -OH, -OCH ₃ or -OCH, or R ^14a and R ^14b together form a ketone group; and R ¹⁵ and R ¹⁸ are independently selected from -H or -OH, or R ¹⁵ and R ¹⁸ are both keto and ring E is a p-benzoquinone ring; provided that at least one of rings A or E is a p-benzoquinone ring; the reactions according to the method include: a) conversion of the quinone system from ring E into a phenolic system; b) converting the quinone system from ring A into a phenolic system; c) conversion of the phenol system from ring A into a methylenedioxyphenol ring; d) forming a bridged spiro ring system of formula (IV), (V), (VI) or (VII) through positions 1 and 4 in ring B; e) obtaining derivatives; f) conversion of a compound wherein R ¹² is -CH 3 or into a group in which R ¹² is selected from -H and -CH ₂ CH ₃ ; and g) converting the 21-cyano substituent to a substituent -OH or -H, as appropriate; to obtain the desired compound of formula (XXIb). A method according to claim 30, wherein in the compound of formula (XXIIb), R ³ is OH, R ^14a and R ^14b are H, R ¹⁵ is H and R ¹⁸ is OH. 32. The method of claim 30, wherein the compound of formula (XXIIb) is ecteinascidin 743 or ecteinascidin770: 65893 Bl 33. The method of claim 30, wherein the compound of formula (XXXX) is ecteinascidin 743: CH _{3 about} BUT 34. The method of claim 30, wherein the product is of formula (XXIII): wherein R ¹ is a hydrophobic group. The method of claim 35, wherein R 1 is a group -CH ₂ NH-COR ^a , in which R ^a has a linear chain with 5 lengths less than 20 atoms. A method according to claim 36, wherein R ^a has a linear chain of less than 15 atoms in length. 38. The method of claim 37, wherein R ^a has a linear chain of less than 10 atoms in length. A method according to any one of claims 34 to 38, wherein R ⁵ is an acetyloxy group. The method of claim 34, wherein the compound of formula (XXIII) is a phthalascidine of formula (III): wherein: R ¹ is a medium-sized aminomethylene group; R ⁵ is a small size hydroxyl group; R ¹² is -CH ₃ ; _H R ²¹ is hydroxyl or cyano. The method of claim 34, characterized by A method according to claim 25,26 or 27, wherein aa is alanyl. The method of claim 41, wherein the alanyl group is protected by a butoxycarbonyl group. A method according to any one of claims 1 to 42, characterized in that it comprises the following reaction: OMe where R ¹ and R ¹⁸ are as defined in a strong group. claim 1 or 30. 45. A method according to any one of claims 44. The method according to claim 43, characterized in that 1 to 44, characterized in that the R ¹⁸ is a protected hydroxyl, comprises the reaction: 65893 B1 where R ¹ and R ¹⁸ are as defined in claim 1 or 30. 1θ A method according to any one of claims 1 to 45, characterized in that it includes a reaction in which a compound of group R ¹ which is an aminomethylene group is converted to a compound of group R ¹ which is hydroxymethyl $ new group. 47. A compound of formula: wherein R ¹ is a possibly protected or derivative aminomethylene group; and R ¹⁸ is an ether protected hydroxyl group. A compound according to claim 49, characterized in that it has the formula: characterized in that R ¹ is 25 51. Compound of Formula: a protected or derivative aminomethylene group; and R ¹⁸ is an ether protected hydroxyl group. A compound according to claim 47, characterized in that it has the formula: 49. A compound of formula Me OMe wherein R ¹ is a possibly protected or derivative aminomethylene group; and R ¹⁸ is an ether protected hydroxyl group. A compound according to claim 51, characterized in that it has the formula: