FI90427B - N'-alkoxy or N'-aralkyloxy-N6-cyano-griseolinic acid derivatives which can be used as intermediate products - Google Patents

Description

.90427 Intermediate N'-alkoxy or N'-aralkyloxy-N6-cyano-griseolinic acid derivatives Somellanprodukter användbara N'-alkoxy-eller N'-aralkyloxy-N6-cyano-griseolin syraderivat 5 (Separated from the invention is directed to a number of novel griseolinic acid derivatives used as intermediates in the preparation of other compounds.

Griseolinic acid is a nucleoside-type compound comprising an adenine base and two carboxylic acid groups. It was first published in, among other things, European patent publication no. 29 329A, but at that time its structure was not yet known. Its structure was first disclosed in U.S. Pat.

15 4 460 765 (granted to the authors of this application). Subsequently, some derivatives of grise oleic acid were disclosed in U.S. Patent Application Serial No. 664,866, filed October 25, 1984, by the present applicants, and the structure of griseolic acid is also disclosed in this application. Other derivatives of griseolinic acid, in particular dihydrodesoxygriseolinic acid, its salts and esters, are disclosed in U.S. Pat. 734,868, filed May 16, 1985.

In accordance with the recommendations of the International Union of Pure and Applied Chemistry (IUPAC), the compounds of this invention are designated as derivatives of griseolic acid (or dihydrodesoxygriseolic acid), with griseolinic acid as the basic structure. Used. the numbering system is disclosed in U.S. Patent Application 664,866.

30 Griseolinic acid and application no. The grise-oleic acid derivatives of 664 866 and 734 868 and the derivatives of this invention * .. * are capable of inhibiting the activity of phosphodiesterases specific for a number of cyclic nucleotides, such as phosphodiesterases such as the 3 ', 5'-cyclic adenosine monophosphate 35 diesterase (PDE) or 3 ', 5'-cyclic guanosine monophosphate (cGMP) -

• PDEs, and thus are able to add a cyclic nucleotide such as cAMP

or cGMP, levels in the cells of a patient treated with such a compound.

2 90427

It is well known that cAMP, which is widely distributed in animal tissues, acts as a second transmitter of a large number of hormones and mediates the effect of this set of hormones; as a result, the cAMP molecule has many very important physiological and biochemical functions. In addition, it is known to affect or be involved in: cell division, proliferation, and specialization; the systolic system, especially the contraction of the heart (myocardia); blood cell formation and development (hematopoiesis); various functions of the central nervous system; immune reactions; and the release of insulin and histamine. Its concentration in tissues, and thus its effect on these different events, depends on the balance between the cAMP-synthesizing enzyme (i.e., adenylate cyclase) and the cAMP-degrading enzyme cAMP PDE. cAMP An inhibitor of the PDE enzyme increases the level of cAMP in cells and thus can be expected to have numerous medical applications, for example: in the treatment of cardiovascular diseases; as an asthma cure agent; as a smooth muscle trigger; as a psychotropic or neurotropic substance; as an anti-inflammatory agent; in the treatment of cancer; as well as in the treatment of diabetes.

To date, the activities of other cyclic nucleotides, such as cGMP, have not been elucidated in such detail. However, they are believed to have a number of effects that are similar to, but not identical to, the effects of the cAMP molecule. Thus, inhibition of PDE enzymes specific for such other cyclic nucleotides likewise results in medicinal effects similar to those resulting from inhibition of the cAMP PDE enzyme. Once the activities of these other cyclic nucleotides have been elucidated, there is a need to find inhibitors of PDE enzymes associated with these other nucleotides that have greater specificity for one of the other 30 nucleotides, rather than a cAMP PDE; in fact, the development of such inhibitors may even facilitate or encourage research into such other cyclic nucleotides.

In addition to griseolinic acid and its derivatives, other compounds known to inhibit the PDE enzymes of cAMP and cGMP include, for example, papaverine, dipyridamole, and some compounds similar to nucleic acid bases such as theophylline or M & B 22 948. [Kukovetz et al., Naunyn-Schmiedeberg's Arch. Pharmakol., 310, 129 (1979)].

Within the scope of this invention, a number of compounds have been found, such as griseolinic acid and dihydrodesoxygriseolinic acid, which are not themselves pharmaceutically active, but which are valuable intermediates in the preparation of other griseolinic acid and dihydrodesoxygriseolinic acid derivatives having griseolinic acid and dihydrodeoxyglycine and dihydrodes. Surprisingly, some of these compounds, which can be prepared from the intermediates of the invention, have a higher activity against the cGMP PDE than against the cAMP PDE.

The aim of the applicant's FI application 861655 was to provide derivatives of griseolinic acid-15, as well as its salts and esters, which are new compounds.

The specific aim of Applicant's FI application 861655 was to provide derivatives of griseolinic acid capable of inhibiting the activity of 20 cyclic nucleotide degrading PDE enzymes, such as cAMP PDE or cGMP PDE.

"The compounds of this invention are compounds of formula (XL): 25 ° C HCl

Yx '> Γ. 30 »8« 5 β

Si 35 R * (X1) 4 90427 R1 and R2 are independently selected from the group consisting of a hydrogen atom and groups of the formula -OR9; R 3 and R * are independently selected from the group consisting of carbamoyl groups and carboxy groups and lower alkoxycarbonyl groups, and these lower alkoxycarbonyl groups are optionally substituted with a phenyl or benzhydryl group; R 5 and R 6 both represent a hydrogen atom, or form an additional carbon-carbon bond between the carbon atoms to which they are attached; R9 represents a hydrogen atom, a lower alkanoyl group, a benzyl group or a benzoyl group; and R30 represents a lower alkyl group or an aralkyl group; as well as pharmaceutically acceptable salts and esters of these compounds.

The present invention provides compounds for:: treating phosphodiesterase imbalances *; animals suffering from disorders, in particular mammals (such as humans) - by administering to said animal a phosphodiesterase inhibitor, the phosphodiesterase inhibitor being selected from the group consisting of the compounds of formula (I) of FI application 861655, and pharmaceutically acceptable salts and esters thereof.

The compounds of this invention can be prepared by administering a compound of formula 30 (XXXIX) 35 5 90427

q NHCN

'Co 10 R 2, (XXXIX) [wherein R 2, R 2, R 2, R 2, R 8 and R 9 are as defined above]; react with a lower alkyl halide or aralkyl halide in the presence of an organic base; and i) optionally, any of the above groups R 1, R 2, R 31 R * 1 R 5, R 6 is replaced with any other group according to the definitions of said groups.

As used herein, the term "aryl group", either as such or as part of a larger group (e.g., an aralkyl group), means a carbon-cyclic aryl group having preferably 6-14, more preferably 6-10, ring-forming carbon atoms (e.g., phenyl or 1- or 2-naphthyl). ), which may be substituted or unsubstituted. If this group is substituted, the substituents are preferably selected from the following groups: C 1 -C 4 alkyl groups, C 1 -C 4 alkoxy groups, hydroxy groups, halogen atoms, nitro groups, amino groups, C 1 -C 4 alkylamino groups, dialkylamino groups in which each alkyl moiety is C 1 -C 3, C 1 -C 3 haloalkyl groups, C 2 -C 7 alkoxycarbonyl groups, aryl groups (which are themselves defined herein, preferably phenyl groups, substituted or unsubstituted, but if substituted, preferably non-aryl groups) and cyano groups.

When R 1 or R 2 represents a group of the formula -OR 9, and R 9 is as defined above, then -OR 9 represents a hydroxy group, a C 1-6 alkoxy group or a protected hydroxy group.

6 90427

If the -OR 9 group represented by R 1 or R 2 is said C 1 -C 6 alkoxy group, it may be a straight-chain or branched group exemplified by methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec.butoxy, t-butoxy, pentyloxy and hexyl-5-hydroxy groups.

If the group -ORR9 represented by R1, R2 or R® represents said substituted ethoxy group, this ethoxy group may then have one or more, preferably 1-3, substituents selected from the following groups: C1-C4-alkoxy groups, C C 1 -C 4 alkyl groups, halogen atoms, C 1 -C 4 alkylselenyl and arylselenyl groups (wherein the aryl moiety is as defined above). Examples of such groups include a 1-ethoxyethoxy group, a 1-methyl-1-methoxyethoxy group, a 1-isopropoxyethoxy group, a 2,2,2-trichloroethoxy group and a 2-15 phenylselenylethoxy group.

The compounds of formula (XL) comprise two carboxy groups, so that they can form mono- or di-salts as well as mono- or di-esters. In the case of di-salts and di-esters, the cationic moieties of the salts or the alcohol moieties of the esters may be the same or a different group. In practice, however, it is most easy to prepare, in particular, di-salts and di-barriers in which the two cationic moieties or the two alcohol moieties are: the same.

25 There are no particular restrictions on the nature of the alcoholic moiety of the ester. Any of the esters conventionally formed for this type of compound can also be formed with the compounds of this invention. Examples of such esters are: C 1 -C 6 alkyl esters, in particular methyl, ethyl, propyl, isopropyl, butyl, isobutyl and t-butyl esters; aralkyl esters, especially benzyl, p-nitrobenzyl, o-nitrobenzyl, triphenylmethyl, bis (o-nitrophenyl) methyl, 9-anthrylmethyl, 2,4,6-trimethylbenzyl, p-bromobenzyl -, p-methoxybenzyl, piperonyl and benzhydryl esters; O-C6-haloalkyl esters which may have one or more halo-35 gene atoms (such as chlorine, bromine, fluorine or iodine atoms) up to complete perhalogenation, for example 2,2,2-trichloro-7 90427 riethyl, 2 -chloroethyl, 2-bromoethyl, 2-fluoroethyl, 2-iodoethyl and 2,2-dibromoethyl esters; alkoxymethyl esters, wherein the alkoxy moiety is preferably C 1 -C 3, such as methoxymethyl, ethoxymethyl, propoxymethyl, isopropoxymethyl and butoxymethyl esters; aliphatic acyloxyalkyl esters (especially acyloxymethyl and acyloxyethyl esters) such as acetoxymethyl, propionyloxymethyl, butyryloxymethyl, pivaloyloxymethyl, 1-acetoxyethyl, 1-propionyloxyethyl, 1-butyryloxyethyloxyethyloxyethyl ; (C 1 -C 4 (-) alkyl) oxycarbonyloxyethyl esters such as 1-methoxy-10-carbarbonyloxyethyl, 1-ethoxycarbonyloxyethyl, 1-propoxycarbonyloxyethyl, 1-isopropoxycarbonyloxyethyl, 1-butoxycarbonyloxyethoxyethoxy and 1-isobutyloxyethyl and 1-isobutoxyethoxy such as phthalidyl esters, heterocyclyl methyl esters (wherein the heterocyclic group is preferably as defined for R9), for example (5-methyl-2-oxo-1,3-dioxolen-4-yl) methyl esters, and esters which are easily hydrolyzed which class comprises certain esters from the classes listed above [for example, aliphatic acyloxyalkyl esters, lower alkoxycarbonyloxyethyl esters, (5-methyl-2-oxo-1,3-dioxolen-4-yl) methyl esters and phthalidyl esters].

No particular limitation is imposed on the nature of the cations which form a salt with the compounds of the invention. Preferred salts are, for example, alkali metal salts (such as sodium or potassium salts) or alkaline earth metal salts (such as calcium salts).

The compounds of the invention also form acid addition salts. The nature of the acid used to form these salts is not critical. Examples of such acids include: inorganic acids such as hydrochloric acid, sulfuric acid and phosphoric acid; organic carboxylic acids such as acetic acid, oxalic acid, maleic acid, succinic acid, citric acid, tartaric acid, fumaric acid, lauric acid, stearic acid and palmitic acid; as well as certain organic sulfonic acids such as methanesulfonic acid, benzenesulfonic acid and g-toluenesulfonic acid.

35 8 90427 The molecules of the compounds of this invention have many asymmetric carbon atoms and can therefore exist as numerous steroisomers. Both individual isolated isomers and mixtures of these isomers are within the scope of this invention. As a natural product, griseolinic acid is a single isomer in which both the 2 'and 7' carbon atoms are in the R-configuration; compounds made from griseolinic acid may retain this same configuration, or may have the opposite configuration on one or more asymmetric carbon atoms. For example, when R 1 is a group or atom other than hydrogen, the configuration of the compounds at the 2 'position may be e or β. When R 2 represents a group or atom other than hydrogen, the 7 'position of the configuration may be R 5, E, or S.

The most preferred class of compounds of this invention are: 1. Compounds of formula (I) wherein R 3 and R * are independently selected from the group consisting of carboxy groups, carbamoyl groups, C 2 -C 5 alkoxycarbonyl groups, (5-methyl-2-oxo -1,3-dioxolen-4-yl) methoxycarbonyl groups, phthalidyloxycarbonyl groups and C2-C5 alkoxycarbonyl groups having at least one substituent selected from aryl groups, aliphatic carboxylic (C1-C6 acyloxy groups) and C1-C4 alkyloxy groups .

2. Compounds of formula (XL) wherein: R 1 and R 2 are independently selected from the group consisting of a hydrogen atom and a hydroxy group.

3. Compounds as defined in 2 above, wherein: R3 represents a carboxy group, a C2-C5 alkoxycarbonyl group, a (5-methyl-2-oxo-1,3-dioxolen-4-yl) methoxycarbonyl group, a phthalidyloxycarbonyl group or a C2-C5- an alkoxycarbonyl group having 1 or 2 substituents selected from C 1 -C 6 alkoxycarbonyloxy groups and C 6 -C 10 90427 carbocyclic aryl groups, the latter aryl groups being unsubstituted; and R 4 represents a carbamoyl group, or any group defined above in connection with R 3.

Examples of compounds of the invention are shown in the following formula (1-5), wherein the substituents are as defined in Table 5, respectively. Hereinafter, the compounds of the invention will be referred to in these tables by the numbers assigned to the compounds, if necessary.

The abbreviations used in the tables have the following meanings: 15 Ac acetyl

Boz benzoyl

Bu butyl

Bz benzyl

Bzh benzhydryl 20 Dox (5-methyl-2-oxo-1,3-dioxolen-4-yl) methyl

Not ethyl

Hx hexyl

We methyl

Piv pivaloyl - 25 Pn pentyl 30 '' 35 10 90427

HH

χ ».J“ '! l tumr = y_l

S ^ OOC-O »OH OH

n 90427

Table 5

Compound

5 No R3a RAb W

175 H H OMe 176 H H OBz 178 Me Me OMe 10 179 Me Me OBz 181 Bzh Bzh OMe 182 Bzh Bzh OBz 15 The following of the compounds listed above are preferred: compounds numbered 175, 176, 178, 179.

More preferred compounds are those numbered 175 and 176.

The compounds of FI application 861655 can be prepared as described in the following reaction schemes.

These reaction schemes show how the intermediates of the invention are prepared (step 30) and how they are used as drugs. . : 25.

i2 90427 NH2 f iX> Ä ¢ 0 0 _ HOOK 0 OH R2300C-C ° OH21 · OH OH21 · (Δ) (X> J phase ** | or they 2

"XrA N" VS

CNX / Άκ / “O AisHl> rtoc>»> ts, '"»' -y.

“Mm 11.1

vaiheJ

I phase 5 '

Co 0c ">

AksTh '

Η23000 - (^ 0 OR50 R OOC \ R2i. ° R

H (XIV) IXII) i3 90427 cV> (yiV) step 6 N step 7t U H X n r22 °° c ^ H7

R2300C- \ 0 OR50 H

(XV) j nh2 step 8 R22 ° 0sri- / Η00ι: νΓ ”Ο r23ooc — on50 hooc - ('o oh

H H

(XVII (0) 14 90 427

OH

r6 r5 n 0 R6 R5 „and ^ ^ c iH V £ IiL ^ R ^ OOC— \ ® OR ^ 1, R ^ OOC — OR ^ 1, R2a R2a (XVIII (XVIII) xl, T> Χ2χ ^ Η'χ ^ Η step 10 "" R6 r5 h 22ooc j— \ y • n K n'-1 IXIX) R ^ 00C— (0 gR2i · R2a / \ step 12 step 11 /

'I

rV> rY> x ^ k ^ 'q6 d5 q6 r5

LuK 14 / N

H0 ° C / \ / hooc / \ 7

H00C - ('0'X oh HOOC - /' ^ OH

V9 r29 (xxi (xxi)

X

is 90427 n ^ VNv I I y

vhihe 13 yl'N ^ 'V

(xvi in ——- R6 R5 g r22o ° cJ \! r23ooc- (s'0 OR21, 'r2s ixxii) step 15 j step 11 * H * ("VS 1jC"> 1.6 ^ O phylo R22 ° QC } \ HOOcK y

R2300C-? A ° OR? 1 * H ° 0C_r29 ° ° H

(XXII1I

(XXIV1

B

βΛ / "χ x \ X) phase 15 r6 r5 HOOC- (0 OH R29 K (XXV) 16 90 427 V1 rV \ (XV,„ I Ä g d5

„I_L / ° xJ KXVII

R 0QCV \ _ / R23OOC- \ 0 OR21 R2a

stage IQ

y 'I I)> Y2> ^ H ^' N / „6 r5 I« VII1

H00C ^ “W

Hm - \ * OH

R ^ y 17 90 427 nh2 nh2 I iC '> Γχ>

H step 19 H

t t K t I / O.

r22 ° oc z 'T_ / r22ooc h-r_7

R23 0 0C— (0 0R2t R23OOC-OH

V »V3

IXXVI1!) (XXI XI

18 90427 ΗΠ2 n | «2 | τ \ rV> ^ H step 20 ^ R6 R5 0 R5 f5„

R2200C) -R2200C, 2 \ -Y

R2300C- \ 0 OH r23 ° 0C — v7 ° 0R

r29 r29 (XXIX) (ΠΧ |.% JL.

YY> step 21 _ step 22 - R6 R5 - ~

R2200C J ~ X_J

R 23 ° C - 13 ° OH

r 29 (XXXI) 30 Ϊ * Η2 ^ '"' rS" "Ys Ύ>« ™ H 'R6 R5 0 R6 R5 o

R31 ° ocsMly r31 00C ^ mO

fl3200C — OH R3200C - ('' '9 0H

? 9 xr29 R (ΧΧΧΠΙ R (XXXIII) i9 90 427 NH2

HO-N ^ JPA

steps R step 25 (XXXII). , - «- R6 R5 0

R31 ° C

r32 ° 0C and 29 ° H

9 9 (XXXIV) nh2 R33nh N «'' Sr'-A.

1 I% il /

HS ^ N ^ H step2g R S N

r6 r5 R6 R5.

r31ooc J ~ X_J ^ oc ^ HQ1

R3200C- \ 0 OH R3Z °° C — vT'0 0H

> 29 R29 (xxxY) (xxxvii R55 rV> hs ^ N ^ * step 27 R6 R5 · n r31ooc> 0

R3200C— (® ^ OH

Xr29 (XXXVII) 20 90 427 t \ © ©

> -NH XU

Xx> step 20 c _c step 29 (XXX) - Rb Rb Q -

R2200C I

R23OOC- ° OH

V9

(XXXVIIII

0 HHCN R% | CH

• “VV” r \ k, X / step 30 R6 R5 „" 6 o r22ooc J ~ \ _y r22 °° c tO-v R23ooe-T ° oh R23oocX ^ o oh r29 R29 (XXXIX1 (XL) 2i 90427

NHOR30 NHOH

rV> rV> h2h ^ n ^ n Hzn ^ k ^ »7 step 31 step3 2 IXU" * "R6 R5 o - R6 R5 0.

R31OOC ^ y \ / R31O0Cn / ~ J

R3200C - / ^ ° OH R32OOC- <; 0 OH

R ^ r29 K (XLI) H (XLII) step 33 NHOR30 ι, ίί: ΙχΓ "\ JL · I .___ / step3 £.

R6 R5 R22OOC.V \ / ”H2

R% c-C9 ° OH liS

(XUIII h2n ^ n ^ n R6 R5 0 R3100C) ^ ^ «Μοα-Λο'οΗ ixliv) 22 9 0 4 2 7 R33 * OR30 NHOR3® rV> rV '> step 35 Η2Ν · ^ Ν' ^ 'Ν Η2Ν ^ ϊΗ ^^ Ν.

R6 R5 0 + R6 R5 0 R2200C.i \! R2300C- \ 1 ° OH R23O0C - (^ 0 oh 1.23 IXL! I "r29 IXIVI1 / nh2 - i 1 /

HjH ^ H ^ i R6 fl5 Q

R% cJ \! -Ο-Λ ° "οη r29 (XLIV) 23 90 427

* 2 OH

N Ipv step 37 HO ^^ N **

IXLIVl -— + "U

R6 R5 0 R6 R5 R31oocyHf y r31ooc J V ^ y '

r3Z ° 0C — Cq ° 0H R3200C — 0 OH

/ (XLVIII) R (XLIX)

X

Rys

HO'X ^ N ^ " 'N

R5 R5 0 {L)

r31 ° 0Cn / K-J

* 3C-C ° OH

035 rVN> HO ^ N ^ - "step 39

- Φ RS 0, UI

R% C— (O OH R29 24 9 0 4 2 7 R33 ^ OR30

Xr rV> (XLVI, V! ^ I ^ N *! Ϋϋϋ- R R 0 r% c / 1 Vy R3200C — OR37 r38 I LII! HNOR30 NH2

"^ Srv

\ A, X.>, Λ XH> I H R36-N N "

H c, - step 12 Xu step U

ft \ ~ fto _ r31 ° 0C '/ \ 7 R31oocy \ / R3200C- (0 0R37ä R 32OOC - (^ 0 0R37a r3 a (uin fl38a vest

'OH OH

„36_ν / ^ νΧ ^ Ν H2N ^ tH '^' N

H step h R6 R5 o ~ R6 R5 n r31 °° c Jryi

R ^ OOCyO QR37a HOOC- (0 OH

3 (LV) r29 (LVI! 25- 90 427 HNOR30 1 L> r39 = N-At ^ H7

step L5 step AR

(XLIII) -, c -— R6 r5

-22 ° 0Cn ^ W

R2300C—? B ° OH

D29

K (LVID

] H2 OH

Γι ') rrs r39 = n / ^ K' ^ 'N r39_h-A% (| .A. || / r6 „5 vi! IfiL„ 6 „5„ L_W "/ Le.

R R31OOCvV \ y

R2300C-Q ° oh R3200C- (u ^ OH

R <™> R29 (11X1

OH

X &

lie 1.8 H2N

- R * r5 -, LVI1

H00C v / ~ t / mK \? W

26 90 427 r37nor? ° ιΛ- «, step 49 Η2ΛΛ« va1h (XLIII) - -li

lt6 R5 Q

r22 °° Cs ^ 1Γ / R2300C — ςΌ ^ OR37 n36 R (IXI) R37NH r37nh N | Τ \ va i he 51 R6 R5 0 ~ r6 r5 r31 °° CV ^ ~ WR% CJ ~~ \ j R3700C_g0 0R37> Ac -N >> a ILXI1) R * (LXIII) step 52 -— (XLVUi) 27.90 427 ~ ί rY \, m step53 R35-N ^ K ^ «R6 R5„ r3i °° cj ~ w R3200C ^ {v 0R37a o38a (LXIV) r35 rV> β36_ν / ^ ν / ^ ν /

step SU

R6 R5 0

R% tJ ~ W

R ^ 00 - (^ 0 OR373 p38a (LXV) 28 9 0 4 2 7

Τ2 X

Ιι> νΛτ \ IXL1V) νί2ϋ11 + Χ Ν f ΐ, Κ «δ Φ 0« 3100C-M_7 r3100CsM ^ 7 R32 ° 0C — d23 ° 0H Rk00C - (^ 0 oh R ILXVI | Ra (LXV1I) // I35 / rV > / X ^ S ^ S7 s & ihe 56 "R6 R5.

R3i °° cv41y

r3200C - ("O OH

r29 (IX1X) 29 9 0 4 2 7

R6 R5 n A R6 R5 n A

H00C - ('0' ^ oh HOOC— 'OR50

OH. OH

ILXXI (UXI) R6 R5 a ΐϋίΐϋ R2200C y 59 R23ooc> 0> -jR5o

OH

uxxm

fl6 R5 0 A

R2200C) y y step 60 B23OOC — C 0 OR50 OR51 (ΙΧΧΙΙΠ

R ”R- r a R5 R- n A

step 61 mCJ ~ ty

R2300C- \ ° or50 H00C- \ 0 OH

R52 R52

ILXXIVI 'ILXXVI

30 9 0 4 2 7 l> ° 4 ILXXII) vlitle 6L r22 ° 0CsV- \ 17 v3ihe 63t

r2300c_7 ^ 0 ^ Jh50 OR53 (LXXVII

f f5 o f R6 R5 „A

R2200c 7 \ y step 64 R 2200C) Y J

R2300C- (S'0 oh R23OOC- \ ° OR51 OR53 0R53 Π-ΧΧνί!) (LXXVtni R5 R5 Q Δ step 55 R ^ QQC / \ / step 65 r23ooc - \ '0' ^ '\, OR33 r52

(LXXIX I

R5 n A

ηοοο ο- ^ ^ Λ

OH K

(LXXX) 3ΐ 90 42 7 T "

νη2 R5t | H

L 1V> vaihe> step 67 R6 R5 „n6 j ^ o R3100CJ— \ / R310DCV '/ ~~ y 7 r32ooc \ r 32ooc- \ lij Λ,' r27 V Nr27 Rl (LXXXI) (LXXXIt)« H · * 32 . 90427

In the above formulas: R 1 -R 12 and A are as defined above; R2a represents a hydrogen atom or a protected hydroxy group; R22 and R23 are the same or different group, and each represents a carboxy protecting group; R2 * represents a hydroxy protecting group; X represents a halogen atom; R27 represents a hydrogen atom, a hydroxy group or a protected hydroxy group; R28 represents a lower alkyl group, for example any lower alkyl group defined in connection with R12; XI and X2 are the same or a different group, and each represents a halo-20 gene atom; R29 represents a hydrogen atom or a hydroxy group; Y1 and Y2 are the same or a different group, and each represents a hydrogen atom, a hydroxy group, a mercapto group, an amino group, a protected amino group or a group of the formula -SR28; R30 represents an alkyl group or an aralkyl group; R31 and R32 are the same or different and each represents a hydrogen atom or a carboxy protecting group; R 33 represents a hydrogen atom, a C 1 -C 6 alkyl group, an aralkyl group, an aliphatic C 1 -C 20 acyl group or an aromatic acyl group, for example 35 groups defined above in connection with the group R 10; 33 90 427 R33a represents a C1-C6 alkyl group or an aralkyl group; R 3 * represents a hydrogen atom, a C 1 -C 6 alkyl group, a C 1 -C 20 aliphatic aliphatic group, an aromatic acyl group or an aralkyl group, for example a group as defined in connection with R 5, provided that both R 33 and R 3A do not represent a hydrogen atom; R 1 represents a group of the formula -OR 9, -NR 10 RU or -SR 9, for example a group as defined for R 7; R2 represents a hydrogen atom, an aliphatic C1-C20 acyl group, an aromatic acyl group or a trialkylsilyl group; R37 represents an aliphatic C 1 -C 10 acyl group, an aromatic acyl group or a trialkylsilyl group; R37a is any group or hydrogen atom as defined for R37; R38 represents a hydrogen atom, an aliphatic Cx-Cao acyloxy group, an aromatic acyloxy group or a trialkylsilyloxy group; R38a represents any group or hydroxy group defined in connection with R38; R39 represents a substituted methylene group (for example, as defined in connection with a substituted methylene group represented by R10 and R11); Z represents a hydroxy group or an amino group; R50 represents an aliphatic acyl group, an aromatic acyl group or a trialkylsilyl group; R 51 represents a C 1 -C 6 alkylsulfonyl group, a C 1 -C 6 fluoroalkylsulfonyl group or an arylsulfonyl group; 34. 90427 R52 represents a hydrogen atom or a halogen atom; R53 represents a tetrahydropyranyl group or a trialkylsilyl group (as defined, for example, in connection with the group R9); R5A represents a C1-C8 alkyl group; and R55 represents a hydroxy group, a halogen atom, a hydrazine group, a substituted amino group, an amino group protected by a substituted methylene group, or a group represented by the formulas -OR9 or -SR9 defined above.

In these reaction schemes, either griseolinic acid of formula (A) or dihydrodeoxy-15 griseolinic acid of formula (B) is used as the starting material, with the following formulas (A) and (B): 20 NH 2 25 3 8 'mXlQr <' '

HOOC — y 0 0H S1 OH

UI

35 35 9 Π 4 9 7 ΚΗ2

5 I S

10 ο HOOC ί \ / HOOC- '0 i „15 (Β)

For the sake of clarity, the numbering system (A) of griseolinic acid shown above shows a numbering system used throughout this publication.

As already mentioned above, griseolinic acid is a known compound disclosed, for example, in European Patent Publication No. 29,329 or 25 U.S. Patent No. 4,460,765. Dihydrodeoxyigriseoleic acid was disclosed in European Patent Publication No. 0162715, published after the filing date of the present application. Both griseolinic acid and - 'dihydrodeoxygriseolinic acid' can be produced by culturing appropriate

Microorganisms of the genus Streptomyces, in particular Streptomyces gri-: 30 seoaurantiacus strain SANK 63479 (deposited October 9, 1979 in the strain collection "Fermentation Research Institute", Agency of Industrial Science and Technology, Japan, from which it is available under the accession number. roll FERM-P5223, and on 22 October 1980 to the stock collection "Agricultu

ral Research Service ", Peoria, USA, where it is available under accession number 35 la NRRL 12314). Details of Streptomvces priseoaurantiacus strain SANK

36 9 0 4 2 7 63479 are disclosed in European Patent Publication No. 29,329A and U.S. Patent No. 4,460,765.

Step 1 In this step, griseolinic acid (A) is reacted to protect its hydroxy and carboxy groups. The nature of the reaction used will depend on the nature of the protecting groups desired, and subsequent reactions are intended only to illustrate this step. Of course, it will be appreciated that any other reaction known in the art for protecting carboxy or hydroxy groups may be used at this stage.

To protect the carboxy groups, griseolinic acid (A) is preferably reacted with a diazo compound, for example diazomethane or diphenyldiazomethane, or with a triazine compound, in particular a p-tolyltriazene derivative such as N-methyl-p-tolyltriazene. The reaction is preferably carried out in the presence of a solvent, the nature of which is not critical, provided that it does not adversely affect the reaction, and provided that the starting materials can be dissolved in this solvent, at least to some extent. Examples of suitable solvents include: ketones such as acetone; ethers such as tetrahydrofuran; amides such as dimethylformamide; and mixtures comprising water and one or more of the aforementioned organic solvents. The reaction can be carried out over a wide range of temperatures, and in particular the reaction temperature chosen is not critical, although it is generally preferred that the reaction be carried out at a temperature of -20 to + 50 ° C. The time required for the reaction will vary, depending on many factors, notably the nature of the starting materials and the reaction temperature; however, for example, at room temperature, the reaction usually takes a period of 1 to 24 hours.

The hydroxy groups of griseolic acid are similarly protected, either before or after protection of the carboxy groups. This can be accomplished, for example, by reacting griseolinic acid, or griseolin-35-acid protected with carboxy groups, with an acid halide such as acetyl chloride or benzoyl bromide, or an acid anhydride such as acetic anhydride in the presence of 37,90427 bases. The reaction is preferably carried out in the presence of a solvent, the nature of which is not critical, provided that it does not adversely affect the reaction. In general, it is preferred to use pyridine, which also acts as a base. The reaction can be carried out over a wide range of temperatures, and in particular the chosen reaction temperature is not critical; however, in general, it is preferable to carry out the reaction at a temperature of from -20 ° C to room temperature. The time required for the reaction will vary considerably, depending on many factors, notably the nature of the reagents and the reaction temperature; however, when the reaction 10 is carried out in said temperature range, a period of 1 to 15 hours is usually sufficient.

If desired, the amino group at the 6-position of the griseolic acid is likewise converted to a hydroxy group. This is preferably accomplished by reacting griseolin-15 acid, or protected griseolinic acid, with a nitric acid salt, e.g., sodium nitrite, in the presence of acetic acid. The reaction is preferably carried out in the presence of a solvent whose solvent nature is not critical, and thus an aqueous solution of acetic acid is usually preferably used. If the starting material is sparingly soluble, then an acetic acid buffer having a pH of about 4 can be used. This reaction can be carried out over a wide range of temperatures, although usually the reaction is carried out at room temperature for convenience. The time required for the reaction may vary considerably, depending on many factors, notably the nature of the reagents and the temperature employed in the reaction; However, at said temperature, the period is 15 to 50 hours. -. usually sufficient.

Step 2 In this step, the hydrogen halide is attached to both sides of the double bond in the grise-oleic acid derivative of formula (X) to give a compound of formula (XI). The nature of the hydrogen halide H-X used in this reaction depends on the nature of the halogen atom X to be attached, but it is generally preferred to use hydrochloric acid, hydrobromic acid or hydroiodic acid in the reaction. The reaction is preferably carried out in the presence of a solvent, the nature of which is not critical, provided that it does not adversely affect the reaction and that the starting materials can be dissolved therein, at least to some extent. An example of a suitable solvent is an organic acid such as acetic acid. The reaction can be carried out over a wide range of temperatures, for example from 0 to 100 ° C, although in general the reaction can be carried out easily either at a temperature from 0 ° C to room temperature or by heating the reaction mixture at a temperature in the range of 80 to 100 ° C. . The time required for the reaction will vary considerably depending on many factors, notably the reaction temperature and the nature of the solvent and reagents, but a period of from 1 to 72 hours will usually suffice.

Step 3 In this step, the halogen atom X attached in Step 2 at the 4 'position of the compound of formula (XI) is removed by reduction. The reducing agent is preferably either a trisubstituted tin hydride such as tributyltin hydride in an aromatic hydrocarbon solvent such as benzene, or zinc powder, in which case a lower aliphatic acid such as acetic acid or an alcohol such as methanol is preferably used as the solvent. If trisubstituted stannous hydride is used as the reducing agent, the reaction is preferably carried out at about the boiling point of the solvent, and the time required for the reaction is usually 2 to 10 hours. If zinc powder is used as the reducing agent, then the reaction is carried out at a temperature of 25 to 100 ° C, and the reaction time is usually 2 to 20 hours.

Step 4 In this step, griseolinic acid of formula (A) is converted to a derivative of formula (XIII).

The hydroxy group in the 2 'position can be protected by reacting griseolinic acid with an acylating agent, for example also as described in step 1 35 to attach the acyl group R50, and then the carboxy groups of griseolinic acid can also be protected. If desired, the 6-amino group of griseolinic acid can be converted to a hydroxy group using the appropriate reactions described in Step 1, and this conversion reaction can be carried out either before or after the above-mentioned protection steps.

The hydroxy group at the 7 'position is converted to the sulfonyloxy group by -OR51 by reacting the compound with a sulfonylating agent such as lower alkylsulfonyl halides (such as methanesulfonyl chloride), arylsulfonyl halides (such as E-toluenesulfonylsulfonyl) sulfonyl sulfonyl sulfide, or fluorinated lower alkyl. The reaction is preferably carried out in the presence of an acid scavenger, the function of which is to remove the hydrogen halide released in the reaction medium from the reaction medium. Suitable acid scavengers include pyridine and dimethylaminopyridine. Reaction 15 is preferably carried out in the presence of a solvent, the nature of which is not critical, provided that it does not adversely affect the reaction in any way. Suitable solvents are halogenated hydrocarbons, in particular halogenated aliphatic hydrocarbons, such as methylene chloride or chloroform. The reaction can be carried out over a wide temperature range of 20 ° C, and no restrictions are placed on the exact temperature selected; however, for convenience, the reaction is generally carried out at a temperature of -10 ° C to room temperature. The time required for the reaction will vary considerably, depending on many factors, notably the reaction temperature and the nature of the reagents; however, a period of 1-20 hours 25 is usually sufficient.

Step 5 In this step, the sulfonyloxy group at the 7'-30 position of the compound of formula (XIII) is replaced by a halogen atom (by reacting said compound with an anhydrous lithium halide in an acidic amide such as dimethylformamide) and then a hydrogen atom (with a reducing agent).

The first reaction is preferably carried out by the method described in step 60 below.

40 9 0 4 2 7

The reducing agent is preferably zinc in aqueous acetic acid, in which case the aqueous acetic acid itself may act as a reaction solvent. The reaction can be carried out over a wide range of temperatures, for example, from 0 to 150 ° C, and the time required for the reaction to vary considerably is usually 1 to 10 hours.

Steps 6 and 7 In these steps, a compound of formula (XIV) 10 prepared as described in Step 5 is first reacted with a hydrogen halide to give a compound of formula (XV), and then this compound of formula (XV) is reduced to give a compound of formula (XV). XVI). The reactions carried out in these steps are exactly the same as those described in connection with steps 2 and 3 above, and can be carried out using the same reagents and the same reaction conditions.

Step 8 In this step, the carboxy and hydroxy groups of the dihydrodeoxygriseolinic acid of formula (B) are protected, and optionally its 6-amino group is converted to a hydroxy group. These reactions are exactly the same as those described in Step 1 and can be carried out the same. . using the same reagents under the same reaction conditions.

25

Step 9 The starting material used in this step, a compound of formula (XVII), may be any of the compounds of formula (XII) or (XVI) prepared as described above. At this point, the nucleic acid base at the 1 'position is converted to an alkanoyloxy group by reacting a compound of formula (XVII) with (i) sulfuric acid or trifluoromethanesulfonic acid, (ii) a lower carboxylic acid, and (iii) its anhydride. The reaction is preferably carried out in the presence of a solvent, the nature of which is not critical, provided that it does not adversely affect the reaction in any way and that reagents 4i 90427 are soluble therein, even to some extent. The most preferred solvents are lower aliphatic carboxylic acids, which also act as reagents. The reaction can be carried out over a wide range of temperatures, for example, from 0 to 100 ° C; in general, the reaction is preferably carried out either at a temperature of 0 ° C to room temperature or by heating the reaction mixture at a temperature of 80 to 100 ° C. The time required for the reaction varies depending on many factors, notably the solvent used in the reaction and the reaction temperature, but usually a period of from 1 to 72 hours is sufficient.

10

Step 10 In this step, a sugar derivative (XVIII) prepared as described in Step 9 is subjected to a glycosidation reaction by reacting compound 15 (XVIII) with a trimethylsilylated nucleic acid base in the presence of a Lewis acid catalyst, using conventional methods described, for example, in S. Suz. ., [Chem. Pharm. Bull., 18, 172 (1970] or H. Vorbrueggen et al., [Chem. Ber., 106, 3039 (197)] to give a compound of formula (XIX).

The nucleic acid base used is a purine derivative corresponding to the portion of the nucleic acid base desired for incorporation and may have been trimethylsilylated by conventional methods described in A.E. Pierce et al., [Silylation of Organic Compounds, 434 (1968)].

.- .: 25

The nature of the Lewis acid used in the glycosidation reaction is not determined. . no special requirements, and examples of suitable Lewis acids are tin tetrachloride or trimethylsilyl trifluoromethanesulfonate. The reaction is preferably carried out in the presence of a solvent, the nature of which is not critical, provided that it has no adverse effect on the reaction. Suitable solvents are polar solvents such as 1,2-dichloroethane or acetonitrile. The reaction can take place over a wide range of temperatures, and the precise reaction temperature selected is not particularly critical. Usually, the reaction is preferably carried out at a temperature of from room temperature to 150 ° C. The time required for the reaction may vary considerably, depending on many factors, notably the nature of the reagents and the reaction temperature; however, at temperatures in said range, a period of 24 to 72 hours is usually sufficient.

If the substituent in the nucleic acid base to be silylated is a hydroxy group, a mercapto group, an alkylthio group, a halogen atom or a hydrogen atom, the nucleic acid base is silylated directly. If the nucleic acid base has an amino substituent, this amino substituent should preferably be protected by acylation first, before silylation. The ratio of yields of 7- or 9-substituted isomers varies depending on the reaction temperature. For example, glycosidation at room temperature using bistrimethylsilyl-N2-acetylguanine yields more of a 7-substituted compound than a 9-substituted compound. On the other hand, the same reaction at 80 ° C yields more of the 9-isomer than the 7-isomer.

Step 11 In this step, the acyl group or groups at the 2 'position, and optionally also at the 7' position 20, are removed to obtain free hydroxy groups, and the carboxy protecting groups are also removed. The compound of formula (XIX) is preferably dissolved in a dilute aqueous alkaline solution, for example a 0.1-1 N aqueous alkaline solution, preferably a solution of sodium hydroxide or potassium hydroxide, and the mixture is allowed to stand to remove the protecting groups. This reaction can be carried out over a wide range of temperatures, but is usually conveniently carried out at about room temperature. The time required for the reaction may vary considerably, but at said temperature a period of 1 to 10 hours is usually sufficient.

30

Step 12 In this step, a compound of formula (XIX) prepared as described in Step 10 is reacted to selectively convert a 6-halogen atom 35 to an optionally substituted mercapto group comprising an alkyl group, or to a hydrogen atom or a hydroxy group or an amino group or a protected group. an amino group. The reaction used can be carried out, for example, in the manner described in L.B. Townsend, [Nucleic Acid Chemistry, 2,693 (1978)], by reacting a compound of formula (XIX) with various nucleophiles such as sodium bisulfide, sodium alkane thiolate, sodium hydroxide, alcoholic ammonia, methylamine or dimethylamine. The reaction is preferably carried out in the presence of a solvent, the nature of which is not critical, provided that it does not adversely affect the reaction. Suitable solvents are lower alcohols such as methanol or ethanol. The reaction can be carried out over a wide range of temperatures, although in general the reaction is most conveniently carried out at a temperature of from room temperature to 150 ° C, preferably in a closed tube. The time required for the reaction may vary considerably, depending on many factors, notably the nature of the nucleophile and the reaction temperature; however, in said temperature range, a period of 2 to 20 hours is usually sufficient.

This reaction usually results in the deprotection of the protecting groups.

Step 13 This is a glycosidation reaction, and is essentially the same as the reaction described in Step · 10, and can be carried out using the same reaction conditions to give a compound of formula (XXII).

25

Step 14 In this step, the hydroxy protecting groups and carboxy protecting groups are removed, and this can be accomplished essentially as described in Step 11, using the same reagents and reaction conditions. If Y1 represents a protected amino group, for example an acetylamino group, the protecting group can be removed by a hydrolysis reaction. This is done with an alkali, for example a methanolic solution of ammonia (preferably a methanolic solution of ammonia of about 20%). Reaction 35 can be carried out over a wide range of temperatures, although the reaction can generally be carried out most conveniently at about room temperature. The time required for the reaction ω 90427 can vary considerably; however, at the proposed temperature, a period of 24-50 hours is usually sufficient.

Step 15 In this step, a sugar derivative of formula (XVIII) is reacted with a trimethylsilylated nucleic acid base according to the glycosidation reaction described in Step 10 to give a compound of formula (XXIV). The reaction conditions and reagents are essentially the same as in the reaction described in Step 10.

False 16 In this step, the protecting groups are removed to give a compound of formula (XXV) 15. This reaction is essentially the same as the reaction described in Step 11, and can be carried out using the same reagents and reaction conditions.

Step 17 In this step, a sugar derivative of formula (XVIII) is subjected to a glycosidation reaction by reacting it with a trimethyllylated nucleic acid base to give a compound of formula (XXVI). The reagents and reaction conditions are substantially as described in Step 10 above.

Step 18 In this step, the compound of formula (XXVI) prepared in Step 17 is reacted to remove the protecting groups to give the compound of formula (XXVII). The reactions required are substantially in accordance with the reactions set forth in Step 11 and can be carried out using the same reagents and reaction conditions.

35

Step 19 45.90427 In this step, a compound of formula (XXVIII), which may be any of the compounds (X), (XII), (XIV) or 5 (XVI) prepared as described above, is protected by 2'-hydroxy and 7'- the protecting groups are removed from the hydroxy groups (where RZa is a protected hydroxy group) to give a compound of formula (XXIX). This can be carried out by the hydrolyl reaction described in step 11 and can be carried out under the same conditions and using the same reagents, although care must be taken not to hydrolyze the carboxy-protecting groups at the same time. If they hydrolyze, they can be recovered by the esterification method described in Step 1.

Step 20 In this step, a compound of formula (XXIX) is converted to a N-oxide of formula (XXX).

This reaction is preferably carried out by reacting a compound of formula (XXIX) with a peroxide, preferably in a solvent. The nature of the solvent to be used in this reaction is not particularly critical, provided, however, that it does not adversely affect the reaction and that the reagents can be dissolved therein, at least to some extent. Examples of preferred solvents include lower alcohols such as methanol and ethanol. Likewise, there are no particular restrictions on the nature of the peroxide to be used, and examples include hydrogen peroxide and organic peroxides such as m-chloroperbenzoic acid. Preferably, organic peracids such as m-chloroperbenzoic acid are used. The reaction can be carried out over a wide range of temperatures, although usually the reaction is preferably carried out at a temperature in the range of 0 to 60 ° C, preferably at about room temperature. The time required for the reaction may vary considerably, depending on many factors, notably the reaction temperature and the nature of the reagents; however, in the proposed temperature range, a period of 35 to 48 hours is usually sufficient.

Step 21 46 90427 In this step, the N1 oxide is converted to an N1 alkoxy or N1 aralkyloxy compound of formula (XXXI) by reacting the Nx oxide (XXX) with a lower alkyl halide or aralkyl halide in the presence of an organic base, and preferably in the presence of a solvent. The nature of the solvent to be employed is not particularly critical, provided that it has no adverse effect on the reaction and that it can be dissolved, at least to some extent. Suitable solvents include acid amides such as dimethylformamide or dimethylacetamide. Suitable organic bases include trialkylamines such as triethylamine. No particular limitation is imposed on the nature of the halide, provided, however, that it is capable of alkylating or aralkylating a hydroxy group. Suitable lower alkyl halides include methyl iodide, and suitable lower Aral-15 alkyl halides include, for example, benzyl bromide and p-nitrobenzyl bromide. The reaction can be carried out over a wide range of temperatures, although the reaction can usually be carried out easily at temperatures in the range of 0 to 100 ° C, preferably at about room temperature. The time required for the reaction may vary considerably, depending on many factors, notably the nature of the reagents and the reaction temperature; however, a period of 1-20 hours is usually sufficient.

Step 22 In this step, a compound of formula (XXXII) is prepared by a reaction in which a ring of a N-alkoxy or N1-aralkyloxy compound of formula (XXXI) is opened, followed by removal of the resulting formyl group.

These reactions are preferably carried out by reacting a compound of formula (XXXI) with an aqueous alkaline solution. Suitable aqueous aqueous solutions include aqueous solutions of alkali metal hydroxides such as sodium hydroxide. The preferred solution is 1.5 N aqueous sodium hydroxide. The reaction can be carried out over a wide range of temperatures, for example from -10 to + 100 ° C. The time required for the reaction may vary considerably, depending on many factors, notably the nature and concentration of the reagents and the reaction temperature; however, the proposed reagents 4? Using 90427 temperatures and temperatures, a period of 6 minutes to 10 days is usually necessary. This reaction also removes the carboxy protecting groups R22 and R23. Thus, if these groups are needed, they must then be reconnected by the esterification reaction described in Step 1 5. In general, in the later steps, it is preferred that these carboxy groups be protected, so both R31 and R32 should represent a carboxy protecting group.

Step 23 In this step, the compound of formula (XXXII) is reduced to give the compound of formula (XXXIII) and, if desired, the carboxy protecting groups are removed and / or converted to amides and / or converted to salts and / or other carboxy protecting groups.

15

Reduction of a compound of formula (XXXII) to remove the -OR30 group is preferably accomplished by treating the compound with an active metal in the presence of a solvent. The nature of the solvent is not particularly critical, provided that it has no adverse effect on the reaction and that it can dissolve the starting materials at least to some extent. Suitable solvents include mixtures of a dilute aqueous solution of an acid, such as dilute aqueous hydrochloric acid, and a water-miscible organic solvent, such as acetone. The preferred solvent is a mixture comprising equal volumes of 1N aqueous hydrochloric acid and acetone. There are also no particular restrictions on the nature of the active metal used, provided that it can be used in a reduction reaction in which the acid acts as a proton donor. Raney nickel is preferably used as the active metal. This reaction can be carried out over a wide range of temperatures, although in general the reaction may be most conveniently carried out at temperatures in the range 0-50 ° C. The time required for the reaction may vary considerably, depending on many factors, notably the nature of the reagents and the reaction temperature; however, under the conditions proposed above, a period of 1 to 10 hours is usually sufficient.

35 48 9 0 4 2 7

The protecting groups may be removed if desired, the nature of the reaction used for removal depending on the nature of the protecting groups, as is well known in the art.

If a lower alkyl group is used as the carboxy protecting group, it can be removed by treating the compound with a base, especially an early methoxide hydroxide such as sodium hydroxide. It is preferable to use an aqueous solution of an alkali metal hydroxide, for example, a 1 N aqueous solution of sodium hydroxide. The reaction is preferably carried out in the presence of a solvent, the nature of which is not critical to the invention, provided, however, that it does not adversely affect the reaction. In general, an aqueous solution is effective. The reaction can be carried out over a wide range of temperatures, although in general the reaction can be carried out effortlessly at about room temperature. The time required for the reaction may vary considerably, depending on many factors, notably the nature of the reagents and the reaction temperature; however, under the conditions proposed above, a period of 1 to 15 hours is usually sufficient.

If the carboxy protecting group is a diaryl-substituted methyl group such as a diphenylmethyl group (i.e., a benzhydryl group), it is preferably removed under acidic conditions in the presence of a solvent. The nature of the solvent used in this reaction is not particularly critical, provided that it does not adversely affect the reaction. Examples of suitable solvents include aromatic hydrocarbons and aromatic ethers such as anisole. The acid is preferably a fluorinated organic acid such as trifluoroacetic acid. The reaction can be carried out over a wide range of temperatures, although the reaction can generally be carried out effortlessly at about room temperature. The time required for the reaction may vary considerably, but a period of from 30 minutes to 10 hours will usually suffice.

If the carboxy protecting group is an aralkyl group or a lower haloalkyl group, it is preferably removed by reduction. Preferred reducing agents include: in the case of lower haloalkyl groups, aqueous zinc / acetic acid; and in the case of aralkyl groups, hydrogen and 49 90427 a catalyst (such as carbon-bonded palladium or platinum) or an alkali metal sulfide (such as potassium sulfide or sodium sulfide). The reaction is preferably carried out in the presence of a solvent, the nature of which is not critical, provided that it does not adversely affect the reaction. Examples of suitable solvents include; alcohols such as methanol or ethanol; ethers such as tetrahydrofuran or dioxane; fatty acids such as acetic acid; or mixtures comprising one or more of these organic solvents and water. The time required for the reaction is not particularly critical, although usually the reaction 10 can be carried out preferably at a temperature of from 0 ° C to room temperature. At such a temperature, the reaction usually requires a period of from 5 minutes to 12 hours.

If the carboxy protecting group is an alkoxymethyl group, it can be removed by treating the compound with an acid in a solvent. Preferred acids are; hydrochloric acid; a mixture of acetic acid and sulfuric acid; or a mixture of p-toluenesulfonic acid and acetic acid. There is no particular restriction on the nature of the solvent to be employed in this reaction, provided that it has no adverse effect on the reaction. Suitable solvents include: alcohols such as methanol or ethanol; ethers such as tetrahydrofuran or dioxane; as well as a mixture comprising. . one or more of the above solvents and water. No particular limitation is imposed on the temperature of the reaction, although the reaction can usually be carried out effortlessly at a temperature in the range of 0 to 50 ° C, in which case a period of 10 minutes to 18 hours is usually sufficient.

If the carboxy protecting group is removed by treating the compound with aqueous ammonia, it will usually result in the conversion of the carboxy groups at the 8 'and 7' positions to carbamoyl groups.

If desired, the free carboxylic acid can be converted into a salt, for example an alkali metal salt, by conventional methods. For example, in a suitable reaction, the acid is dissolved in a mixture of water and a water-miscible organic solvent such as ethyl acetate, to which is added an aqueous solution of the appropriate alkali metal carbonate or bicarbonate so 90427 (such as potassium carbonate or sodium bicarbonate) at the appropriate temperature (e.g. 0 ° C to room temperature). after which the pH is adjusted so that the salt can separate by precipitation (e.g. to approximately 7).

5

If desired, the resulting hydrochloric or carboxylic acid compound can be converted to an ester in which the carboxy groups are protected with readily hydrolyzable protecting groups in vivo. The salt or acid is first dissolved in a suitable solvent, for example: an ether such as tetrahydrofuran; or a polar solvent such as dimethylformamide, dimethylsulfoxide, hexamethylphosphoric triamide or triethylphosphate. It is then reacted with at least 2 equivalents of a base, which may be an organic base (such as triethylamine or dicyclohexylamine), an alkali metal hydride (such as sodium hydride) or an alkali metal carbonate or bicarbonate (such as sodium carbonate, potassium carbonate or sodium bicarbonate), and reacting the resulting salt with a lower aliphatic acyloxymethyl halide (such as acetoxymethyl chloride or propionyloxymethyl bromide), a lower alkoxycarbonyloxyethyl halide (such as 1-methoxycarbonyloxyethyl chloride, or 1-ethoxycarbonyloxy) halodialcarbonyloxy) phenyloxyloxyethyloxyethyl 1,3-dioxolen-4-yl) methyl halide. The reaction is preferably carried out in the presence of a solvent, the nature of which is not critical, provided that it does not adversely affect the reaction. Suitable solvents are, for example, the polar solvents mentioned above. The reaction can be carried out over a wide range of temperatures, although in general the reaction can be carried out readily at temperatures in the range 0-100 ° C. The time required for the reaction can vary considerably; however, under the conditions proposed above, a period of 30 minutes to 10 hours is usually sufficient. The desired step described above can also be applied if desired in steps 24, 25 and 25, as well as in each step of steps 27, 32, 33, 34, 36, 37, 38, 50 and 54.

35

Step 24 5i 90427 In this step, a compound of formula (XXXII) prepared as described in Step 22 is catalytically reduced to remove the alkyl or aralkyl-5 group R 30 to give a hydroximino compound of formula (XXXIV). This reaction is preferably carried out in the presence of a solvent, the nature of which is not critical, provided that it does not adversely affect the reaction and that it is capable of dissolving the starting materials, even to some extent. Organic carboxylic acids are preferred, especially fatty acids such as acetic acid. There are no particular restrictions on the nature of the catalyst, and any catalyst commonly used in catalytic reduction processes can be used as well in this reaction. However, platinum oxide, carbon-bonded platinum or carbon-bonded palladium, preferably activated carbon adsorbed palladium chloride, are preferably used in the present reaction. The reaction is preferably carried out by stirring a mixture of a catalyst and a compound of formula (XXXII) in a selected solvent under a hydrogen atmosphere. The reaction can take place over a wide range of temperatures, and the particular temperature selected is not critical. In general, the reaction can be carried out effortlessly at temperatures in the range of 5 to 60 ° C. The time required for the reaction may vary considerably, depending on many factors, such as the nature of the compound (XXXII) and the reaction conditions, in particular the reaction temperature; however, under the conditions proposed above, a period of 1 to 10 hours is usually sufficient. If desired, the optional procedure described in step 23 may be performed with the resulting compound.

Step 25 In this step, a compound of formula (XXXIV) prepared as described in Step 24 is reacted with carbon disulfide to form a ring to give a compound of formula (XXXV). This reaction is preferably carried out in the presence of a solvent, the nature of which, however, is not critical, provided that it does not adversely affect the reaction and that it is capable of dissolving the starting materials, at least to some extent. However, a mixture of a weakly basic solvent (such as pyridine) and a lower alcohol (such as methanol) is preferably used in the reaction, with the volumes of the two components preferably being equal in this mixture. The amount of carbon disulfide used is preferably about one-fifth of the amount of solvent. The reaction can be carried out over a wide range of temperatures, and usually the reaction can be carried out conveniently at temperatures in the range of 0 to 150 ° C, preferably at about 80 ° C. The time required for the reaction will vary depending on many factors, notably the nature of the starting materials and the reaction temperature; however, under the conditions proposed above, a period of 10 to 30 hours is usually sufficient. If desired, the optional procedure described in step 23 may be performed with the resulting compound.

Step 26 In this step, a mercapto compound of formula (XXXV) is reacted with an alkyl halide, aralkyl halide, aliphatic acyl halide or aromatic acyl halide of formula R33X and / or R3 * X to form such an alkyl, aralkyl or acyl group. or a 6-amino group. The reaction is preferably carried out in the presence of a solvent, the nature of which is not critical, provided that it does not adversely affect the reaction and that it is capable of dissolving the starting materials at least to some extent. However, water or a lower alcohol (such as methanol) or a mixture of water and such alcohol is preferably used in the reaction. The reaction is preferably carried out in the presence of an acid scavenger, the nature of which is not critical to the acid scavenger, provided that it is capable of removing the hydrogen halide HX formed in the reaction. Suitable acid scavengers include, for example: inorganic bases, especially alkali metal hydroxides such as sodium hydroxide or potassium hydroxide; and organic bases, especially organic amines, preferably trialkylamines such as triethylamine. In general, an aqueous solution of an early metal hydroxide such as sodium hydroxide is preferably used in the reaction. The reagent R33X or R34X may be: a lower alkyl halide such as methyl iodide; An aralkyl halide such as benzyl bromide; a lower aliphatic acyl halide such as acetyl chloride; or an aromatic acyl halide, 53 9 0 4 2 7 such as benzoyl bromide. The reaction can be carried out over a wide range of temperatures, although usually the reaction is carried out preferably at temperatures in the range of 0 to 80 ° C, more preferably at about room temperature. The time required for the reaction may vary considerably, depending on many factors, notably the nature of the reagents and the reaction temperature; however, under the conditions proposed above, a period of 1 to 10 hours is usually sufficient.

If the reaction is carried out using an alkyl halide or an aralkyl-10-halide, then preferably the 2-position mercapto group is alkylated or aralkylated. On the other hand, if the reagent is an aliphatic acyl halide or an aromatic acyl halide, then the 6-amino group is primarily acylated. Thus, the same or a different group may be attached to the compound in the form of R33 and R3 *.

15

If desired, the optional procedure described in step 23 may also be performed with the resulting compound.

Step 27 In this step, the 6-amino group of the compound of formula (XXXV) is converted to a hydroxy group and, if desired, this hydroxy group is halogenated to replace it with a halogen atom; if desired, this halogen atom may then be replaced by a hydrazine group, a substituted amino group, an amino group protected by a substituted methylene group, or a group of the formula -OR9 or -SR9. In addition, if desired, the reaction associated with the optional steps described in Step 23 may also be carried out in each step and / or, if desired, in the last step, a mercapto protecting group, an amino protecting group or a hydroxy protecting group may be removed.

The conversion of the amino group to the hydroxy group is carried out as described in the same reaction of Step 1, and can be carried out using the same reaction conditions and the same reagents.

35 5 * 90 427

Halogenation in which the hydroxy group is replaced by a halogen atom can be carried out using conventional halogenating agents capable of converting a hydroxy group in a heterocyclic compound into a halogen atom. Examples are phosphorus oxyhalides such as phospho-rioxychloride or phosphorus oxybromide; as well as thionyl halides such as thionyl chloride; of these, phosphorus oxychloride is preferably used. The reaction is preferably carried out in the presence of an acid scavenger, for example; an acid amide such as dimethylformamide; an aromatic tertiary amine such as diethylaniline or dimethylaniline; or in the presence of a lower trialkylamine such as triethylamine.

The reaction is preferably carried out in the presence of a solvent, the nature of which is not critical, provided that it does not interfere with the reaction, and in general, an excess of the halogenating agent itself is preferably used as the solvent; alternatively, an excess of a halogenating agent may be used in the reaction in admixture with another organic solvent, such as: an ester such as ethyl acetate; or a halogenated hydrocarbon, preferably a halogenated aliphatic hydrocarbon such as methylene chloride. The reaction can take place over a wide range of temperatures, and the precise temperature selected is not particularly critical to the reaction; in general, the reaction is preferably carried out at the boiling point of the solvent used. The time required for the reaction may vary considerably, depending on many factors, notably the reaction time. the nature of the genes and the reaction temperature; however, under the conditions proposed above, a period of 10 minutes to 5 hours is usually sufficient for the reaction.

Thereafter, if desired, the resulting 6-position halogen atom can be converted to one of the above substituents by reacting the compound with a nucleophilic reagent belonging to one of the following classes: mono- and di-alkylamines, such as methylamine or dimethylamine; amines protected by a substituted methylene group, such as benzylideneamine or other amines corresponding to the above-mentioned substituted methyleneimino groups; Hydroxy-substituted lower alkylamines such as 2-hydroxyethylamine; amino-substituted lower alkylamines such as 2-aminoethyl-55 90 427 amine; aralkylamines such as benzylamine; arylamines such as aniline, β-naphthylamine or N-naphthylamine; hydroxylamine; lower alkoxyamines such as methoxyamine; aralkyloxyamines such as benzyloxamine; alkali metal hydroxides such as sodium hydroxy-5 di; alkali metal hydrogen sulfides such as sodium hydrogen sulfide; alkane thiolates of alkali metals such as sodium methanethiolate; as well as alkali metal alkoxides such as sodium methoxide. If the nucleophilic reagent used is an amine, the reaction can be carried out using an excess of amine without adding any acid-binding agent to the reaction mixture. However, if any of the above-mentioned non-amine nucleophilic substances are used in the reaction, the reaction is preferably carried out in the presence of an acid-binding agent which is not itself nucleophilic; suitable are trialkylamines such as triethylamine. The reaction is preferably carried out in the presence of a solvent, the nature of which is not critical, provided that it does not adversely affect the reaction. Examples of suitable solvents are: lower alcohols such as methanol or ethanol; acidic amides such as dimethylformamide or dimethylacetamide; and polar solvents such as trimethyl sulfoxide, hexamethylphosphoric triamide or triethylphosphate. The reaction can take place over a wide range of temperatures, and the precise reaction temperature selected is not particularly critical; in general, the reaction is preferably carried out at a temperature ranging from room temperature to the boiling point of the solvent used. The time required for the reaction may vary considerably, depending on many factors, notably the nature of the reagents and the reaction temperature; however, if the temperature is within the proposed range, then a period of 6-20 hours is usually sufficient.

If desired, protecting groups can be removed at this stage, and the reactions used to remove such protecting groups will vary conventionally, depending on the exact nature of the protecting group.

If a trialkylsilyl group is used as the hydroxy protecting group, it can be removed by treating the compound with a compound capable of producing fluoroanions; an example is tetrabutylammonium fluoride. The reaction is preferably carried out in the presence of a solvent, the nature of which is not particularly critical, provided that it does not adversely affect the reaction. Suitable solvents are, for example, ethers, such as tetrahydrofuran or dioxane. The reaction temperature is not particularly critical, although usually the reaction is preferably carried out at about room temperature. The time required for the reaction can vary considerably, depending on many factors; however, under the proposed conditions, a period of 10-18 hours is usually sufficient.

If an aralkyloxycarbonyl group or an aralkyl group is used as the hydroxy protecting group, it can be removed by contacting the compound with a reducing agent, for example: using a catalyst such as palladium adsorbed on activated carbon or platinum in the presence of hydrogen, preferably at room temperature; or using an alkali metal sulfide such as sodium sulfide or potassium sulfide. The reaction is preferably carried out in the presence of a solvent, the nature of which is not critical, provided that it does not adversely affect the reaction. Examples of suitable solvents are: alcohols such as methanol or ethanol; ethers such as tetrahydrofuran or dioxane; fatty acids such as acetic acid; or mixtures containing one or more of said organic solvents and water. The reaction can be carried out over a wide range of temperatures, although in general it is conveniently carried out at or below room temperature, for example at a temperature in the range of 0 ° C to room temperature. The time required for the reaction may vary considerably, depending on many factors, notably the nature of the starting materials and reducing agents; however, a period of 5 minutes to 12 hours is usually sufficient.

If an aliphatic acyl group, an aromatic acyl group or an alkoxycarbonyl group is used as the hydroxy protecting group, it can be removed by treating the compound with a base in the presence of an aqueous solvent. There is no particular restriction on the nature of the solvent to be employed, and any solvent generally used in the hydrolysis reaction may be used in this reaction. Examples of preferred solvents include water and mixtures containing water and one or more organic solvents, for example: alcohols, 57 90427 such as methanol, ethanol or propanol; or ethers such as tetrahydrofuran or dioxane. Nor are there any particular restrictions on the nature of the base, provided, however, that it does not affect other parts of the molecule. Examples of suitable bases include: carbonates of early metals such as sodium carbonate or potassium carbonate; hydroxides of early metals such as sodium hydroxide or potassium hydroxide; and methanolic ammonia solution. A methanolic solution of ammonia is a more preferred base for deprotection of the nucleic acid base; in other cases, 1 N aqueous sodium hydroxide solution is preferably used. The reaction can take place over a wide range of temperatures, and the particular reaction temperature selected is not critical; however, in general, the reaction is preferably carried out at or below room temperature, for example, at a temperature of from 0 ° C to room temperature. The time required for the reaction may vary considerably, depending on many factors, notably the nature of the starting materials and the reaction temperature; however, under the conditions proposed above, the reaction is usually complete in 1 to 10 hours.

If a tetrahydropyranyl group, a tetrahydrofuranyl group, thio analogs thereof, an alkoxymethyl group or a substituted ethyl group is used as the hydroxy-protecting group, it can be removed by treating the compound with an acid in a solvent. Examples of preferred acids include: hydrochloric acid; mixtures of acetic acid and sulfuric acid; or mixtures of p-toluenesulfonic acid and acetic acid. Reaction 25 is preferably carried out in the presence of a solvent, the nature of which is not critical, provided that it does not adversely affect the reaction. Examples of suitable solvents include: alcohols such as methanol or ethanol; ethers such as tetrahydrofuran or dioxane; or mixtures containing water and one or more of these organic solvents. The reaction can be carried out over a wide range of temperatures, for example, from 0 to 50 ° C. The time required for the reaction may vary considerably, depending on many factors, notably the nature of the starting materials and acids; however, under the conditions proposed above, a period of 10 minutes to 35 hours is usually sufficient.

58 90 427

If an alkenyloxycarbonyl group is used as the hydroxy protecting group, it can be removed by treating the compound with a base. This reaction can be carried out under the same conditions used to remove a hydroxy-protecting group which is an aliphatic acyl group, an aromatic acyl group or an alkoxycarbonyl group. If the hydroxy-protecting group is an allyloxycarbonyl group, it can be removed simply by using palladium and triphenylphosphine or nickel tetracarbonyl, and this reaction has the advantage that very few side reactions occur.

10

In order to remove the hydroxy-protecting groups, the reactions described can be carried out simultaneously with the elimination of carboxy-protecting groups, mercapto-protecting groups and amino-protecting groups, in each case according to the nature of these groups.

After carrying out these reactions, the desired compounds can be recovered from the reaction mixture by conventional means, for example, by recrystallization of the resulting compound by crystallization, preparative thin layer chromatography or column chromatography.

20

If an aliphatic acyl group or an aromatic acyl group is used as the mercapto protecting group, it can be removed by treating the compound with a base. The same reaction conditions used to remove the aliphatic acyl group or the aromatic acyl group used as the hydroxy-protecting group are used in this reaction. Simultaneously with the reaction, elimination of a carboxy-protecting group, a hydroxy-protecting group or an amino-protecting group can be carried out.

After carrying out the reaction, the desired compounds can, if desired, be recovered from the reaction mixture by conventional means, and, for example, purified by recrystallization, preparative thin layer chromatography or column chromatography.

If an aliphatic acyl group, an aromatic acyl group or a substituted methylene group is used as the amino-protecting group, it can be removed by treating the compound with a base. This reaction 59 90427 uses the same reagents and reaction conditions used to remove the aliphatic acyl groups and aromatic acyl groups used as the hydroxy protecting group. Thus, other protecting groups can be removed simultaneously.

5

If a trialkylsilyl group is used as the amino protecting group, it can be removed by the same reactions used to remove the trialkylsilyl group used as the hydroxy protecting group.

Elimination of a carboxy protecting group, a hydroxy protecting group or a mercapto protecting group can be carried out simultaneously with the reaction used to remove these amino protecting groups. After carrying out the reactions, the desired compounds can be recovered from the reaction mixture, if necessary, by conventional means, and can be purified, if desired, by recrystallization by preparative thin layer chromatography or column chromatography.

The order in which the hydroxy protecting groups, carboxy protecting groups, amino protecting groups and mercapto protecting groups are removed is not critical and the removal of these groups can be performed in any order, either sequentially or simultaneously.

Step 28 In this step, an N-oxide of formula (XXX) prepared as described in Step 20 is converted to a polycyclic compound of formula (XXXVIII) by reacting it with a cyanogen halide such as cyanogen bromide. This reaction is preferably carried out in the presence of a solvent, the nature of which is not critical, provided, however, that it does not adversely affect the reaction and that it is capable of dissolving the starting materials, at least to some extent. Examples of suitable solvents are: lower alcohols, such as methanol or ethanol. The reaction temperature is not particularly critical, although usually the reaction is preferably carried out at temperatures in the range of -10 to + 50 ° C, preferably at about room temperature. The time required for the reaction may vary considerably depending on many of the factors used, in particular the nature of the reagents and the reaction temperature; however, under the conditions proposed above, a period of 1 to 10 hours is usually sufficient.

Step 29 In this reaction, a polycyclic compound of formula (XXXVIII) is converted to a N-cyano-t-oxide of formula (XXXIX) by reacting it with a basic compound in a solvent. The nature of the solvent used is not particularly critical, provided, however, that it does not adversely affect the reaction and that it is capable of dissolving the starting materials at least to some extent. Examples of preferred solvents are C1-C4 alcohols, such as methanol or ethanol. No particular limitation is imposed on the nature of the basic compound used, although a solvent saturated with ammonia gas, preferably a methanolic ammonia solution, is usually used most readily in the reaction. The reaction can be carried out over a wide range of temperatures, for example from -10 to + 50 ° C, preferably at room temperature. The time required for the reaction may vary considerably, depending on many factors, notably the nature of the starting materials, solvents and basic compounds; however, under the conditions proposed above, a period of 1 to 5 hours is usually sufficient.

Step 30 In this step, the N-oxide (XXXIX) is converted to a N-alkoxy or N-aralkyloxy compound (XL). The reactions used are the same as those described in Step 21 above, and can be carried out using the same reagents and reaction conditions.

30

Step 31 In this step, a compound of formula (XL) prepared as described in Step 30 is converted to a compound of formula (XLI). Reaction 35 is preferably carried out as follows. First, the compound of formula (XL) is treated with an alkali in a solvent at a pH of 12-13 to open its 61 90427 pyrimidine ring (and also to remove the carboxy protecting groups by chance). The pH of the reaction mixture is then adjusted to 7.0 and the mixture is heated to form a ring again, resulting in a compound of formula (XLI) wherein the carboxy groups are unprotected (i.e., where both R31 and R32 are hydrogen). As the alkali in the first step of this reaction, aqueous solutions of alkali metal hydroxides, such as sodium hydroxide or potassium hydroxide, are preferably used. No particular limitation is imposed on the nature of the solvent used, provided, however, that it does not interfere with the reaction and that it is capable of dissolving the starting materials, at least to some extent. However, water or mixtures of water and a lower alcohol (such as methanol or ethanol) are preferably used in the reaction. Both reactions can be carried out over a wide range of temperatures, and the precise temperature selected is not particularly critical. The ring-opening reaction preferably uses approximately room temperature, and at this temperature, at a pH of about 12, the reaction is completely completed, usually in 30 to 60 minutes. The ring-forming reaction is preferably carried out at a temperature of 0 to 15 ° C, and at this temperature the reaction proceeds to completion in a total time of generally 1 to 5 hours.

If desired, after these reactions, the carboxy protecting groups can be reconnected by the esterification reaction described in Step 1.

Step 32 In this step, the alkyl or aralkyl group R30 is removed and replaced with a hydrogen atom. This reaction is the same as the reaction described in Step 24, and can be carried out under these same reaction conditions, using the same reagents. If desired, the optional procedure described in step 23 may also be performed with the resulting compound.

Step 33 35 In this step, a reaction leading to ring opening and regeneration is carried out, which is similar to the reaction described in Step 31, but in which the carboxy-protecting groups remain in place. The reaction is preferably carried out in the presence of a solvent, the nature of which is not particularly critical, provided that it does not adversely affect the reaction and that it is capable of dissolving the starting materials at least to some extent. However, the solvent is preferably such that it is able to maintain the pH of the reaction solution in the range of 6-8, and thus a mixture of a buffer, pH 6-8, and a lower alcohol (such as methanol or ethanol), preferably a buffer, pH, is preferably used in the reaction. 7.0, and a mixture of methanol. The reaction can be carried out over a wide temperature range of -10 ° C, for example 30-150 ° C. The time required for the reaction may vary considerably, depending on many factors, notably the reaction temperature, the pH, and the nature of the starting materials and solvents; however, under the conditions proposed above, a period of 3 to 10 hours is usually sufficient. If desired, the optional procedure described in step 23 can be performed with the resulting compound.

Step 34 In this step, the alkoxy or aralkyloxy group -OR30 is replaced by a hydrogen atom. The reactions used are similar to those described in Step 23 and can be carried out using the same reagents, under the same reaction conditions, to give the desired compound of formula (XLIV). If desired, the optional step 25 described in step 23 may be performed with the resulting compound.

Step 35 In this step, if the carboxy groups 30 of the compound of formula (XLI) are free (i.e., R31 and R32 represent a hydrogen atom), they can be protected by reactions such as those described in Step 1. It is particularly preferred that benzhydryl groups be added to the compound to protect the carboxy groups by reacting the compound with diphenyldiazomethane. Under the conditions used in this reaction, a protecting group such as a benzhydryl group can also be attached to the 6-amino group to give a compound of formula (XLVI).

63 90427

The reaction is preferably carried out in a solvent which is a mixture of water and a water-miscible organic solvent such as acetone having a pH of 1-2. The reaction is preferably carried out at room temperature and usually requires 1 to 10 hours.

5

Step 36 In this step, the alkoxy or aralkyloxy group -OR30 is removed; the reaction and the reaction conditions used are as described in connection with step 23. If desired, the optional procedure described in step 23 may be performed with the resulting compound.

Step 37 In this step, a compound of formula (XLIV) prepared as described in step 34 or 36 is converted to a monohydroxy compound (XLVIII) or a dihydroxy compound of formula (XLIX) by reacting it with nitrite, for example as described in step 1. If desired, the optional procedure described in step 23 may be performed with the resulting compound.

Step 38 In this step, the 6-amino group of the compound of formula (XLVIII) is converted to a halogen atom by reacting the compound with nitrite. in hydrohalic acid or haloboronic acid, as described in step 1. Preferred hydrohalic acids are, for example, hydroiodic acid, hydrobromic acid or hydrochloric acid. The preferred haloboronic acid is, for example, fluoroboric acid. The reaction can be carried out over a wide range of temperatures, for example from -30 to + 50 ° C. The time required for the reaction may vary considerably, depending on many factors, such as the nature of the starting materials and the reaction temperature; however, a period of 1-20 hours is usually sufficient. If desired, the optional procedure described in step 23 can be performed with the resulting compound.

64 90 427

Step 39 In this step, a group R35 (i.e., a group of the formula -OR9, -NR1R11 or -SR9) is attached to the 6-position of the purine base. The reaction required is similar to that described in Step 5 27, with the optional reactions described in Step 27 also being carried out if desired.

Step 40 In this step, two R36 groups are attached to the 2-amino group and the R37 group is attached to the 2'-hydroxy group using reactions similar to those described in Step 49 below to give a compound of formula (LII).

Steps 41-43 In these steps: first, one of the groups R36 attached in the previous step is removed and the group R33a is removed from the N6 position; at this point, the 2'-hydroxy protecting group and / or the 7'-hydroxy protecting group 20 and / or the carboxy protecting group may be removed. If desired, the carboxy group may be protected as described in Step 1, while the hydroxy groups may be protected as described in Step 40. Next, the alkoxy or aralkyloxy group -OR30 is removed from the N6 position; and converting the 6-amino group to a 6-hydroxy group. These reactions can be carried out as described above using the same reactions and reaction conditions.

Step 44 In this step, all protecting groups are removed, for example, as described in step 30 27.

Step 45 In this step, a compound of formula (XLIII) prepared as described in Step 33 is reacted with dimethylformamide acetal, or an aldehyde such as benzaldehyde, and an organic base 65 9 0 4 2 7 (such as triethylamine) to protect the 2-amino group. with a substituted methylene group such as N, N-dimethylaminomethylene, benzylidene, p-methoxybenzylidene, p-nitrobenzylidene, salicylidene, 5-chlorosalicylidene, diphenylmethyl-5-ethylene or (5-chloro-2-phenylphenyl) phenyl. The reaction is preferably carried out in the presence of a solvent, the nature of which is not critical, provided that it does not adversely affect the reaction and that it is capable of dissolving the starting materials, at least to some extent. Suitable solvents are acid amides such as dimethylformamide or dimethylacetamide. The reaction can be carried out over a wide range of temperatures, although in general the reaction can be carried out readily at room temperature. The time required for the reaction may vary considerably, depending on many factors, notably the nature of the reagents and the reaction temperature; however, at the temperatures proposed above, a period of 15 minutes to 5 hours is usually sufficient.

Step 46 In this step, the alkoxy or aralkyloxy group -OR30 is removed by a procedure similar to that described in Step 23 to give a compound of formula (LVIII).

Step 47 In this step, the 6-amino group is converted to a hydroxy group by giving ... reacting the compound with nitrite in a manner similar to that described in Step 1 to give a compound of formula (LIX).

Step 48 V In this step, the substituted methylene group protecting the 2-amino group on the purine base is removed together with both carboxy protecting groups R31 and R32 in analogy to the procedures described in Step 27 for the optional procedures to give the compound of formula 35 (LVI).

66 9 0 4 2 7

Step 49 In this step, a compound of formula (XLIII) is converted to a compound of formula (LXI) by protecting the nitrogen atom at the 6-position 5 of the purine base and the hydroxy group in the sugar group with the protecting groups defined above. This can be accomplished by acylation, for example, by reacting compound (XLIII) with an acylating agent, such as: an acyl halide such as an aromatic acyl halide (e.g., benzyl chloride) or a lower aliphatic acyl halide (such as acetyl-10 bromide or propionyl chloride); or an acid anhydride, for example an aromatic anhydride (such as benzoic anhydride) or a lower aliphatic acid anhydride (such as acetic anhydride or propionic anhydride). The reaction can be carried out over a wide range of temperatures, for example from -30 to + 100 ° C. The time required for the reaction may vary considerably, depending on many factors, notably the nature of the acylating agent and the reaction temperature; however, in the above conditions, a period of 30 minutes to 50 hours is usually sufficient.

Step 50 In this step, a protected compound of formula (LXI) prepared as described in step 49 is converted to a compound of formula (LXII) analogously to the actual procedure of step 23, and if desired analogously to the optional procedures described in step 27.

"Step 51 In this step, a compound of formula (LXII) is converted to a compound of formula (LXIII) by the method described in step 37, and, if desired, by the optional steps described in step 27.

35

Step S? 67 90427 In this step, a compound of formula (LXIII) prepared as described in step 51 is converted to a compound of formula (XLVIII) 5 by deprotection of the 6-amino group in the purine base and the 2'-hydroxy group in the sugar group using the optional methods described in step 27. Under these conditions, carboxy protecting groups can be removed simultaneously.

Step 53 In this step, a compound of formula (LV) prepared as described in step 43 is converted to a compound of formula (LXIV) comprising a group X at the 6-position of the purine base using a method similar to that described in the second step of step 27, and optionally using step 27 optional methods.

In this step, the group X at the 6-position of the purine base is replaced with R35 using a method similar to that described in step 39, and if desired, the optional methods of step 27.

Step 55 .. In this step, a compound of formula (XLIV) which may have been prepared as described in Step 34 or 36 is converted to a monohalogen or dihalogen compound (IXVI) or (LXVII), respectively, by methods similar to those described in Step 38, and if desired in Step 38. 23 by optional methods such as described 30.

Step 56 In this step, the dihalogen compound (LXVII) is converted to the compound of formula (LXIX) 35 by a method similar to that described in Step 39, and optionally by steps such as described in Step 27, if desired.

Step 57 In this step, only the hydroxy group is acylated at the 2 'position of the sugar moiety of the griseolinic acid derivative of formula (LXX), which derivative may have been prepared by any of the methods described above. The acylation can be accomplished by either: (i) slowly adding a base (such as sodium hydroxide) to the griseolinic acid derivative (LXX), followed by an acylating agent, which may be any of the acylating agents described in the acylation steps above, but preferably an aromatic acyl halide; Benzoyl chloride) to the reaction solution, maintaining its pH in the range of 10-13; or (ii) dissolving the griseolinic acid derivative (LXX) in a buffer solution having a pH of 10-13; after which an acylating agent is added.

20

The reaction is preferably carried out in the presence of a solvent, the nature of which is not critical, provided that it does not adversely affect the reaction and that it is capable of dissolving the reagents at least to some extent. Preferably, a mixture of water and a water-miscible solvent (especially an ester such as ethyl acetate) is used. The reaction can be carried out over a wide range of temperatures, for example from -20 to + 50 ° C. The time required for the reaction may vary considerably, depending on many factors, notably the nature of the reagents; however, a period of 1 to 10 hours is usually sufficient.

30

Step 58 In this step, the free carboxy groups 35 of the griseolic acid derivative of formula (LXXI) (prepared as described in Step 57) are protected by a method similar to the esterification reaction described in Step 1.

Step 59 69 90427 In this step, a compound of formula (LXXII) prepared as described in step 58 is converted to a compound of formula (LXXIII) by sulfonylation. The reaction is carried out by reacting a compound of formula (LXXII) with a lower alkylsulfonyl halide (such as methanesulfonyl chloride), an arylsulfonyl halide (such as n-toluenesulfonyl chloride) or a fluorinated lower alkylsulfonyl halide (such as trifluoromethanesulfone), . The reaction is preferably carried out in the presence of a solvent, the nature of which is not critical, provided that it does not adversely affect the reaction. Examples of suitable solvents are: halogenated hydrocarbons, in particular halogenated aliphatic hydrocarbons, such as methylene chloride or chloro-form. The reaction can be carried out over a wide range of temperatures, although usually the reaction is preferably carried out at a low temperature, for example from -10 ° C to room temperature. The time required for the reaction may vary, depending on many factors, notably the nature of the reagents and the reaction temperature; however, a period of 1-20 hours 20 is usually sufficient.

Step 60 In this step, the - 25 sulfonyloxy group of the compound of formula (LXXIII) at the 7 'position is replaced with a halogen atom (by reacting the compound with anhydrous lithium halide in an acidic amide such as dimethylformamide) and / or a hydrogen atom (reducing the compound with zinc and aqueous acetic acid). above). The former reaction is preferably carried out in the presence of a solvent, the nature of which is not critical, provided that it does not adversely affect the reaction. Suitable solvents are polar solvents such as dimethylformamide, dimethyl sulfoxide, triethyl phosphate or hexamethyl phosphorus triamide. The reaction can be carried out over a wide range of temperatures, for example, from 0 to 150 ° C. The time required for the reaction 35 can vary considerably, depending on many factors, but usually a period of 1 to 10 hours is sufficient.

Step 61 70. 90427 At this stage, the compound of formula (LXXIV) is deprotected by methods appropriate to the nature of the protecting groups. If, for example, a pyranyl group is used as a hydroxy protecting group, it can be removed by treating the compound with an acid such as acetic acid or p-toluenesulfonic acid, preferably pyridine E-toluenesulfonate, in a solvent mixture comprising a lower alkanol (such as ethanol) and a halogenated hydrocarbon (such as methylene). . If a trialkylsilyl group is used as the hydroxy protecting group, it can be removed by treatment with a fluorine anion-forming compound such as tetrabutylammonium fluoride with an ether (such as tetrahydrofuran) as a solvent. The reaction can be carried out over a wide range of temperatures, for example from room temperature to 100 ° C. The time required for the reaction may vary considerably depending on many factors, but usually a period of from 5 to 20 hours is sufficient. The carboxy protecting groups and / or other hydroxy protecting groups can be removed as described in step 27.

20

Step 62 In this step, the hydroxy group in the 7 'position of the compound of formula (LXXII) (prepared as described in Step 58) is protected using a number of methods, depending on the particular protecting group to be attached. If it is desired to protect the group with a pyranyl group, the compound (LXXII) is reacted with a suitable pyran derivative, for example 3,4-dihydro-α-pyran, in the presence of an acid catalyst such as hydrochloric acid. If the group is to be protected with a lower trialkylsilyl group, the compound (LXXII) is reacted with a trialkylsilyl halide such as dimethyl-t-butylsilyl chloride and imidazole. These reactions are preferably carried out in the presence of a solvent, the nature of which is not critical, provided that it does not adversely affect the reaction. Examples of suitable solvents include: halogenated hydrocarbons, especially halogenated aliphatic hydrocarbons such as chloroform; esters such as ethyl acetate 7i 90427 ti; ethers such as dioxane; and acidic amides such as dimethylformamide. The reactions can be carried out over a wide range of temperatures, and in particular the temperature chosen is not critical to the invention. The reaction is usually carried out preferably at about room temperature. The time required for the reaction will vary considerably depending on many factors, notably the nature of the reagents and solvents and the reaction temperature; however, under the conditions proposed above, a period of 1 to 30 hours is usually sufficient.

In this step, the 2'-hydroxy group is deprotected by removing the acyl or silyl group R50 from a compound of formula (IXXVI) to give a compound of formula (LXXVII). The reaction is preferably carried out by contacting the protected compound with an aqueous alkali metal hydroxide solution such as 1 N aqueous sodium hydroxide solution or a 20% v / v solution of ammonia in ethanol. The reaction can be carried out over a wide range of temperatures, for example from -20 to + 50 ° C. The time required for the reaction may vary considerably, depending on many factors, notably the nature of the reagents and the reaction temperature. However, under the conditions proposed above, a period of 10 minutes to 3 hours is usually sufficient.

Step 64-25 In this step, a compound of formula (LXXVII) is sulfonylated analogously to the method described in Step 59, and the reaction can be carried out under the same reaction conditions and using the same reagents to give a compound of formula (IXXVIII).

30

Step 65 In this step, the sulfonyloxy group at the 2 'position of the compound of formula (IXXVIII) is replaced by a halogen atom or a hydrogen atom. The reactions used are similar to those described in step 60 and can be carried out using the same reagents, under the same reaction conditions.

Step 66 In this step, a compound of formula (LXXIX) prepared in Step 65 is deprotected to give a compound of formula (LXXX). The reactions required are similar to those described in Step 61 and can be carried out using the same reagents, under the same reaction conditions.

Since each substitution reaction in steps 60 and 65 involves a Walden inversion, these reactions yield a compound whose etheric configuration is inverse to that of the parent compound. Compounds having a configuration inverse to the configuration of the compound obtained above (i.e., having a naturally occurring configuration) can be prepared, if desired: in the procedure of step 60 or step 65, a lower alkanoyloxy group is attached to the starting material R52; then this lower alkanoyloxy group is removed by the method described in step 20 61; and then the resulting compound is treated again with the procedure of step 59 or the procedure of step 64, respectively.

Step 67 25

In the first part of step 67, griseolinic acid of formula (LXXXI) or a derivative thereof is alkylated or aralkylated with an alkylating or Aralylating agent in an inert solvent. The nature of the solvent is not particularly critical, provided that it does not adversely affect the reaction. Examples of suitable solvents include: alcohols such as methanol, ethanol, isopropanol, butanol, and t-butanol; ethers such as diethyl ether, tetrahydrofuran, dioxane or ethylene glycol dimethyl ether; nitriles such as acetonitrile; amides such as dimethylformamide, dimethylacetamide or hexamethylphosphoric triamide; and 35 sulfoxides such as dimethyl sulfoxide. Amides or sulfoxides are preferably used as the solvent.

73 9 0 4 2 7

The reaction can be carried out over a wide range of temperatures, however, it is preferably carried out at a temperature of 0 to 100 ° C, preferably at a temperature of from room temperature to 70 ° C.

The time required for the reaction may vary widely, depending on many factors, notably the reaction temperature and the nature of the solvent and reagents employed. A period of 30 minutes to 10 days is usually sufficient. If the reaction is carried out, for example, at room temperature, it will generally be completely completed within 1 to 10 days; on the other hand, at a temperature of 70 ° C, it usually wears out in 1 to 20 hours.

The intermediate thus obtained can be recovered from the reaction mixture by evaporating the solvent under reduced pressure, after which the intermediate can be used in the next part of the step without further purification in the same reaction vessel. Alternatively, if desired, this intermediate can be isolated by conventional methods before being used in the next part of the step.

In the second part of step 67, the ring of the intermediate compound is opened, rearranged (rearranged) and resealed, with this reaction targeting the pyrimidine ring and the free amino group.

At this point, the residue obtained in the alkylation or aralkylation reaction is dissolved or suspended in a suitable solvent, and the pH of the resulting solution or suspension is maintained at or adjusted to at least 4, whereby said ring-opening reactions, rearrangement reactions and ring-closing reactions are carried out. The pH used in these reactions is preferably at least 5 and further preferably at least 7.

The pH is maintained at a selected value, for example, by (1) performing the reactions in a buffer solution pre-adjusted to an appropriate value, or (2) standing or heating the residue in excess aqueous alkali metal or alkaline earth metal hydroxide solution or 74 90 427 solution comprising an organic base. in water or a suitable organic solvent.

No particular limitation is imposed on the nature of the buffer solution used, provided that it is capable of maintaining an appropriate pH throughout the reaction. Any conventional buffer solution, for example acetate, phosphate, borate, ammonium bicarbonate, phthalate or citrate buffer, can be used.

Examples of suitable alkali metal and alkaline earth metal hydroxides which can be used in aqueous solution include sodium hydroxide, potassium hydroxide, lithium hydroxide and calcium hydroxide. Examples of suitable organic bases are lower alkylamines, such as monomethylamine, dimethylamine or trimethylamine.

15

In general, the pH of the reaction solution is maintained at a value, preferably in the range of 4-12, although higher pH values may be used.

There are no restrictions on the nature of the solvent used in this reaction, provided that it does not interfere with the reactions. Examples of suitable solvents are: water; alcohols such as methanol, ethanol or propanol; as well as other water-soluble solvents such as acetone, tetrahydrofuran, dioxane, dimethylformamide or dimethyl sulfoxide. Only one such solvent may be used in the reaction, or a mixture of two or more such solvents. In some cases, the organic base may act as a reaction solvent.

The reaction can be carried out over a wide temperature range, for example, from 0 to 150 ° C, preferably from 20 to 100 ° C. The selected heat-30 mode can depend on many factors. For example, heating may be advantageous when the reaction is carried out at a pH in the range of 4 to 10; on the other hand, the reaction usually proceeds satisfactorily at ambient temperature when the pH is 10 or above.

The time required for the reaction may vary considerably depending on many factors, notably the nature of the substrates, the reaction temperature of 75 90427 and the pH, and the nature of the buffer or other medium employed, especially the temperature and pH; however, in the above-mentioned preferred conditions, a period of 5 minutes to 50 hours is usually sufficient. Thereafter, if desired, the optional steps of step 5 can be taken.

After any of the above reactions is complete, the desired product of each step can be separated from the reaction mixture by conventional means. For example, the reaction mixture is washed, if necessary, with water, after which the solvent is distilled off under reduced pressure. The residue can be purified in a variety of ways, such as by recrystallization, or by various chromatographic methods, such as column chromatography or preparative thin layer chromatography, to give the desired compound.

The preparation of medicaments prepared from the compounds of this invention and the preparation of the compounds of the invention are illustrated by the following examples. The preparation of certain starting materials is illustrated in the preparations shown after the examples.

20

The following chain can be mentioned as an example in which the compound (XL) of the invention acts as an intermediate: Example 16 (XL) -> Example 18 -> Example 23 -> Example 25 -> Example 27 -> Example 27 -> Example 28 -> Example 30.

EXAMPLE 1 1 (a) PimethylH-1 * -deadenino-1 '/ 9-acetoxy-4'.5'-dihydro-02'.07*-diazate-tetrylligriseolate 30 2 ml of concentrated sulfuric acid are added to a solution comprising 500 mg Dimethyl 6-desamino-6-hydroxy-4 'β, 5'-dihydro-02', 07'-diacetyl griseolate (prepared as described in Preparation 6) in 100 ml of 4: 1 acetic acid and acetic anhydride (v / v) ), and this mixture was allowed to stand at room temperature for 14 hours under a nitrogen atmosphere. To the reaction mixture was added 15 g of sodium acetate, and the solvent 76 90 427 was evaporated off under reduced pressure. The residue was dissolved in saturated aqueous sodium bicarbonate solution, and this solution was extracted three times with methylene chloride. The methylene chloride extracts were combined and dried over anhydrous magnesium sulfate. The solvent was then evaporated under reduced pressure. The residue was purified by chromatography on a silica gel column, eluting with a 2: 1 (v / v) mixture of cyclohexane and ethyl acetate. Evaporation of the solvent from the second eluting fraction gave 292 mg of the title compound.

10

Nuclear Magnetic Resonance Spectrum (CDCl 3) δ ppm: 2.44 [1H, doublet of doublets, J = 6.4 & 14.7 Hz]; 2.65 [1H, doublet of doublets, J = 14.7 & 3.0 Hz]; 4.98 [1H, doublet of doublets, J = 2.5 & 6.4 Hz]; 5.02 [1H, doublet of doublets, J = 2.9 & 4.0 Hz]; 5.15 [1H, doublet of doublets, J = 4.0 & 2.5 Hz]; 5.72 [1H, singlet]; 6.29 [1H, doublet, J = 2.9 Hz], 20

Mass spectrum (m / e): 474 (M + 43).

Mass spectrum with fast atoms bombardment (m / e): 431 (M +).

25 Kb) Dimethyl-K-deadenino-1'-acetoxy ^ *. 5'-dihydro-O2 '.O7' -diacetyl-ligrise isolate

In the column chromatographic treatment mentioned in Example 1 (a), the first separated fraction was concentrated by evaporation under reduced pressure to give 33 mg of the title compound.

Nuclear Magnetic Resonance Spectrum (CDCl 3) δ ppm: 2.67 [1H, doublet of doublets, J = 7.4 & 15.1 Hz]; 35 2.98 [1H, doublet of doublets, J = 15.1 & 2.9 Hz]; 4.94-4.99 [2H, multiplet]; 90427 5.05-5.07 [1H, multiplet]; 5.61 [1H, singlet]; 6.38 [1H, doublet, J = 4.9 Hz], δ Mass spectrum (m / e): 474 (M + 43).

Mass spectrum with fast atom bombardment (m / e): 431 (Μ *).

1 (c) Dimethyl 1 * -deadenino-1,1'-acetoxy-4,5'-dihydro-02 '.0''-dibent-10 sowligriseolate 2 ml of concentrated sulfuric acid were added to an ice-cooled solution comprising 400 mg of dimethyl- 6-Desamino-6-hydroxy-4 ', 5'-dihydro-O 2', 07'-dibenzoyl griseolate (prepared by a method similar to that described in Preparations 1-3, 5 and 7, but using benzoyl chloride instead of acetic anhydride in Preparation 2) dissolved in 80 to 1 ml of a 4: 1 (v / v) mixture of acetic acid and acetic anhydride, and this mixture was allowed to stand at room temperature for 14 hours. The reaction mixture was then stirred with 15 g of sodium acetate and concentrated by evaporation under reduced pressure. The residue was dissolved in a mixture of methylene chloride and saturated aqueous sodium bicarbonate, and extracted three times with methylene chloride. The methylene chloride extracts were combined and dried over anhydrous magnesium sulfate, after which the solvent was evaporated under reduced pressure. The residue was purified by chromatography on a silica gel column eluting with a 2: 1 (v / v) mixture of cyclohexane and ethyl acetate, and the fractions containing the title compound were concentrated by evaporation under reduced pressure to give 243 mg of the title compound.

Nuclear Magnetic Resonance Spectrum (CDCl 3) δ ppm: 2.50-2.60 [1H, multiplet]; 2.96-3.04 [1H, multiplet]; 5.06-5.16 [2H, multiplet]; 35 5.48-5.52 [1H, multiplet]; 5.88 [1H, singlet]; 78 90 427 6.50-6.58 [1H, multiplet].

Elemental analysis: Calculated for C 27 H 26 O 2: C, 59.78%; H, 4.83%; N, O%.

Found: C, 59.59%; H, 4.80%; N, 0.01%.

Mass spectrum (m / e): 585 (M + 43).

EXAMPLE 2

Dimethyl-6-desamino-6-hydroxy-2-acetylamino-2-dehydro-4,5'-dihydro-15 O2 '. 07'-dibenzowligriseolate 200 mg of dimethyl 1'-deadenino-1'O-acetoxy-4 ', 5'-dihydro-02', 07'-dibenzoylgriseolate (prepared as described in Example 1 (c)) and 200 mg of bistrimethylsilyl Nz -acetylguanine was placed in a two-necked flask under a nitrogen atmosphere. 0.4 ml of trimethylsilyl trifluoromethanesulfonate was added to the solution obtained by dissolving the mixture in 40 ml of 1,2-dichloroethane on ice while keeping cold, after which the mixture was allowed to stand at room temperature for 4 days. The reaction mixture was worked up as described in Example 3. It was purified by chromatography on a silica gel column eluting with 3% (v / v) methanol in methylene chloride to give 54.8 mg of the title compound, which was isolated as the second eluting fraction.

Nuclear Magnetic Resonance Spectrum [(CD3) 2SO] δ ppm: 2.59-2.96 [2H, multiplet]; 5.12-5.32 [1H, multiplet]; 5.32-5.63 [1H, multiplet]; 5.83 [1H, singlet]; 35.11 [1H, doublet of doublets, J = 4.5 & 3.9 Hz]; 6.47 [1H, doublet, J = 4.5 Hz]; η .90427 8.32 [1Η, singlet].

Elemental analysis: Calculated for C 32 H 29 N 5 O 2 .2 / 2H 2 O: C, 56.14%; H, 4.50%; N, 10.23%.

Found: C, 56.28%; H, 4.53%; N, 9.93%.

EXAMPLE 3 10 6-Desamino-6-hydroxy-2-acetylamino-2-dehydro-4 ', 5'-dihydrogriseolinic acid 40 mg dimethyl-6-desamino-6-hydroxy-2-acetylamino-2-dehydro-4' The 5'-15 dihydro-O2 ', 07'-dibenzoyl griseolate (prepared as described in Example 2) was dissolved in 5 ml of 1 N aqueous sodium hydroxide solution under ice-cooling, and this mixture was allowed to stand at room temperature for 4 hours. The mixture was then adjusted to pH 1 with 1 N aqueous hydrochloric acid and subjected to column chromatography using a pre-packed RP-18 column (reverse phase type, Merck) eluted with 5% (v / v) acetonitrile in water. The actual fractions were lyophilized to give 22.0 mg of the title compound.

Nuclear Magnetic Resonance Spectrum (D 2 O) δ ppm: 2.58 [1H, doublet of doublets, J = 6.5 & 15.4 Hz]; 2.76 [1H, doublet of doublets, J = 15.4 & 1.5 Hz]; 4.60-4.75 [3H, multiplet]; Δ 5.10-5.16 [1H, multiplet]; 5.97 [1H, doublet, J = 6.8 Hz]; 8.20 [1H, singlet].

35 80 90427 EXAMPLE 4 6-Desamino-6-hydroxy-2-amino-2-dehydro-4',5'-dihydro-griseolinic acid 5 21 mg 6-desamino-6-hydroxy-2-acetylamino-2-dehydro-4 ', 5'-Dihydrogriseolinic acid (prepared as described in Example 3) was placed in a round bottom flask under a nitrogen atmosphere. To this was added 10 milliliters of methanol containing 20% v / v ammonia. The mixture was then allowed to stand at room temperature for 1 day in a container tightly closed with a stopper. The solvent was evaporated under reduced pressure, and the residue was dissolved in 0.5 N aqueous hydrochloric acid. This solution was subjected to column chromatography using a pre-packed RP-18 column (reverse phase type; Merck). The column was eluted with 3% v / v acetonitrile in water, after which the eluted solution was lyophilized to give 16 mg of the title compound.

Nuclear Magnetic Resonance Spectrum (D 2 O) δ ppm: 2.49 [1H, doublet of doublets, J = 6.3 & 15.5 Hz]; 2.64 [1H, doublet of doublets, J = 15.5 δ 1.5 Hz]; 4.54-4.77 [3H, multiplet]; 5.01-5.05 [1H, multiplet]; 5.89 [1H, doublet, J = 7.3 Hz]; 8.00 [1H, singlet].

25 EXAMPLE 5

N1-oxide of dimethyl griseoleate (starting material of formula XXXIX that can be used to prepare XL 30) 8.14 g of dimethyl griseolate (prepared as described in Preparation 1) was suspended in methanol. To this was added 6.90 g of m-chloroperbenzoic acid, and the mixture was stirred at room temperature for 24 hours. The solvent was evaporated by distillation under reduced pressure. When the solvent was almost removed, 300 ml of diethyl ether was added to the residue, and the lumps in solution 81 90 427 were broken up with a spatula until they were allowed to disappear. The mixture was filtered and the residue was washed with 100 ml of diethyl ether, and dried to give 7.81 g of a white powder. This powder was dissolved in 200 ml of methanol and 300 ml of 5 methylene chloride as completely as possible with the aid of heating, after which the solvent was evaporated under reduced pressure with a suction device so that the volume of the remaining solution was about 50 ml. The resulting crystals were separated by filtration to give 6.31 g of the title compound as white powdery crystals.

Nuclear Magnetic Resonance Spectrum [(CD3) 2SO] δ ppm: 3.68 [3H, singlet]; 3.78 [3H, singlet]; 4.60 [1H, doublet, J = 5.0 Hz]; 4.67 [1H, singlet]; 5.15 [1H, doublet, J = 2.2 Hz]; 5.90 [1H, doublet of doublets, J = 2.2 & 5.0 Hz]; Δ 6.53 [1H, singlet]; 8.53 [1H, singlet]; 8.72 [1H, singlet].

25 EXAMPLE 6

N1-oxide of griseoline sine 1.1 g of N1-oxide of dimethyl griseolate (prepared as described in Example 5 above) was dissolved in 15 ml of 0.5 N aqueous sodium hydroxide solution, and this mixture was allowed to stand at room temperature for 2 hours. The pH of the resulting solution was adjusted to 2.3, after which the solution was subjected to column chromatography using a pre-packed RP-8 column (Merck) and washed with water. The column was eluted with water containing 5% v / v acetonitrile, and the resulting actual fractions were collected and lyophilized to give 300 mg of the title compound as a white powder.

Nuclear Magnetic Resonance Spectrum [(CD3) 2SO] δ ppm: δ 4.53 [1H, singlet]; 4.61 [1H, doublet, J = 5.0 Hz]; 5.15 [1H, doublet, J = 2.2 Hz]; 5.90 [1H, doublet of doublets, J = 2.2 & 5.0 Hz]; Δ 6.52 [1H, singlet]; 8.51 [1H, singlet]; 8.67 [1H, singlet], EXAMPLE 7 15

Dimethyl N1-p-nitrobenzyl hydroxy griseolate 35 ml of dimethylformamide were added to 1.48 g of dimethyl griseolate N1 oxide (prepared as described in Example 5) and 2.27 g of p-nitro-20 benzyl bromide, and this mixture was stirred at room temperature overnight. The solvent was distilled off under reduced pressure, and the residue was crystallized by adding diethyl ether. The mixture was then filtered and the residue was dissolved in a mixture of ethyl acetate and 10% v / v aqueous sodium bicarbonate solution. The separated organic layer was washed with water and dried over anhydrous magnesium sulfate. The desiccant was separated by filtration, and the solvent was removed from the filtrate by distillation, and the residue was then purified by chromatography on a silica gel column eluting with 3% v / v methanol in methylene chloride to give 1.5 g of the title compound.

30

Nuclear Magnetic Resonance Spectrum [(CD3) 2SO] δ ppm: 3.65, 3.73 [total 3H, each singlet]; 4.56 (1H, doublet, J = 5.0 Hz]; 35 4.63 [1H, singlet]; 5.17 [1H, doublet, J = 2.2 Hz]; 83 9 0 4 2 7 5.45 [ 2H, singlet], 5.87 [1H, doublet of doublets, J = 2.2 & 5.0 Hz], 6.41 [1H, singlet], 7.6-8.43 [6H, multiplet].

5 EXAMPLE 8

Dimethyl βτ-benzyloxy griseolate A mixture of 5.35 g of dimethyl griseolate β-oxide (prepared as described in Example 5) and 7.6 ml of benzyl bromide was reacted overnight at room temperature in 90 ml of dimethylformamide. The mixture was then worked up using the procedures described in Example 7. The resulting solution was poured into 1 liter of hexane and 0.5 to 15 liters of diethyl ether with stirring. The resulting powder was filtered to give 5.4 grams of the title compound as a crude powder.

Nuclear Magnetic Resonance Spectrum [(CD3) 2SO] δ ppm: 3.64, 3.71 [total 3H, each singlet]; 4.54 [1H, doublet, J = 5.0 Hz]; 4.63 [1H, singlet]; 5.17 [1H, doublet, J = 2.2 Hz]; Δ 5.33 [2H, singlet]; 5.83 [1H, doublet of doublets, J = 2.2 & 5.0 Hz]; 6.46 [1H, singlet]; 7.3-7.7 [5H, multiplet]; 8.31, 8.39 [total 1H, each singlet].

30 EXAMPLE 9

Dibenzhydryl-1 * -deadenino-1 ', N, N-5-amino-4-CN2-p-nitrobenzyl. amiditol) imidazole-1-yl griseolate 35 84 90427 0.7 g of dimethyl N, N-nitrobenzyloxygriseolate was dissolved in 6.2 ml of 1.5 N aqueous sodium hydroxide solution, and the mixture was stirred at room temperature for 3 days. The reaction mixture was then acidified with 3N aqueous hydrochloric acid, and 20 ml of acetone and 2.5 g of diphenyldiazomethane were added. The mixture was allowed to react to esterify the acid at room temperature for 60 minutes, stirring constantly. At the end of this time, the acetone was removed by distillation, and 30 ml of methylene chloride was added to the mixture. The separated organic layer was washed with 10% w / v aqueous sodium bicarbonate solution and saturated aqueous sodium chloride solution. The solvent was removed by distillation, and the residue was dissolved in 2 ml of acetone. The resulting solution was poured into 300 mL of hexane, stirring constantly. The powder thus obtained was purified by chromatography on a silica gel column eluting with a 1: 1 v / v mixture of cyclohexane and ethyl acetate to give 0.35 g of the title compound.

Nuclear Magnetic Resonance Spectrum [(CD3) 2SO] δ ppm 4.51 [1H, doublet, J = 5.0 Hz]; 4.87 [1H, singlet]; 5.07 [2H, singlet]; 5.25 [1H, doublet, J = 2.2 Hz]; 5.56 [1H, broad singlet]; 6.15 [1H, singlet]; Δ 6.66, 6.72 [total 1H, each singlet]; 7.15-7.4 [21H, multiplet]; 7.66, 8.20 [together 2H, each doublet], 30. EXAMPLE 10

Dibenzhydryl-1 * -deadenino-1 * (β- (5-amino-4-CN 2-benzyloxyamidino) imidazol-1-yl] isolate 35 5.4 g of dimethyl N1-benzyloxygriseolate (prepared as described in Example 8) was dissolved in 53 ml of sodium hydroxide 1, 5 N aqueous 85 9 0 4 2 7 solution, and the mixture was stirred at room temperature for 3 days. The reaction mixture was then acidified with concentrated hydrochloric acid, and then 15 g of diphenyldiazomethane and 100 ml of acetone were added thereto, and it was worked up using the procedures described in Example 9. The resulting mixture was purified by silica gel column chromatography, and the column was eluted with 1% v / v methanol-containing methylene chloride to give 3.9 g of the title compound.

Nuclear Magnetic Resonance Spectrum [(CD3) 2SO] δ ppm δ 4.54 [1H, doublet, J = 5.0 Hz]; 4.90 [1H, singlet]; 4.94 [2H, singlet]; 5.26 [1H, doublet, J = 2.2 Hz]; 5.59 [1H, doublet of doublets, J = 2.2 & 5.0 Hz]; 6.20 [1H, singlet]; 6.69, 6.76 [total 1H, each singlet]; 7.1-7.5 [26H, multiplet].

EXAMPLE 11 1'-Deadenino-1'D- (5-amino-4-amidinoimidazol-1-yl) -greiseic acid 1.05 g of dibenzhydryl-1'-deadenino-1 '- [5-amino-4 - (N2-benzyloxy-25-amidino) imidazol-1-yl] griseolate (prepared as described in Example 10) was dissolved in 60 ml of acetone. To the resulting solution were added 30 ml of a 1 N aqueous hydrochloric acid solution and 6 ml of Raney nickel (W-2), stirring the solution all the time. The mixture was then stirred at room temperature for a further 60 minutes. At the end of this time, the Raney-30 nickel was removed by filtration, the acetone was removed by distillation, and the residue was extracted twice with ethyl acetate. The extract was washed with water, 10% w / v aqueous sodium bicarbonate solution, and then saturated aqueous sodium chloride solution. The extract was then dried over anhydrous magnesium sulfate. The desiccant was removed by filtration, and the solvent was distilled off from the filtrate to give 0.67 g of a residue. This residue was then dissolved in 20 ml of acetone and 20 ml of 86 90 427 water, and the resulting mixture was acidified with concentrated hydrochloric acid. To the mixture was added 1 g of diphenyldiazomethane, and the mixture was stirred at room temperature for 60 minutes. The acetone was removed by distillation, and the residue was extracted twice with ethyl acetate. The extract was washed with 10% w / v aqueous sodium bicarbonate solution and saturated aqueous sodium chloride solution, and dried over anhydrous magnesium sulfate, which was then removed by filtration. The solvent was evaporated from the filtrate and the residue was dissolved in 10 ml of acetone. The resulting mixture was poured into 200 mL of hexane, stirring constantly. The resulting powder was filtered and the residue was purified by chromatography on a silica gel column eluting with 10% v / v methanol in methylene chloride to give 0.15 g of the benzhydryl ester of the title compound.

0.06 g of this benzhydryl ester was dissolved in 0.8 ml of anisole, and 0.8 ml of trifluoroacetic acid was added thereto, keeping the solution cold on ice. To the resulting mixture was added 10 ml of toluene, and the solvent was removed by distillation. To the residue were added 5 ml of acetone and 10 ml of toluene, and the solvent was removed by distillation. This procedure was repeated twice, and then the residue was dissolved in 1 ml of acetone and 20 ml of hexane was added. The resulting powder was filtered and dissolved in 10% w / v aqueous sodium bicarbonate. The pH of the resulting solution was adjusted to 1.9 with 1 N aqueous hydrochloric acid. The solution was purified by column chromatography using 25 prepacked RP-8 columns (reverse phase type, Merck) eluting with water to give 24 mg of the title compound.

Nuclear Magnetic Resonance Spectrum [(CD3) 2SO] δ ppm 4.08 [1H, singlet]; 4.42 [1H, doublet, J = 5.0 Hz]; 4.83 [1H, doublet, J = 2.2 Hz]; 5.44 [1H, doublet of doublets, J = 2.2 & 5.0 Hz]; 6.20 [1H, singlet]; 35 7.67 [1H, singlet].

87 90 427 EXAMPLE 12

D1-benzhydryl-1'-deadenyl-1 * β- [5-amino-4- (N2-hydroxyamidino] imidazole-1-yl] griseolate 1.0 g of dlbenzhydryl-1'-deadenino-1 '[3- [5- amino-4- (N2-benzyloxyamidino) imidazol-1-yl] griseolate (prepared as described in Example 10) was dissolved in 20 ml of acetic acid, followed by the addition of 0.6 g of 10% w / w palladium adsorbed on carbon before The mixture was stirred at room temperature for 2 hours under a stream of hydrogen, the palladium-on-carbon was filtered off and the solvent was removed by distillation, and the residue was dissolved in a mixture of 30 ml of ethyl acetate and 20 ml of 10% w / w aqueous sodium bicarbonate solution. the organic layer was washed with water and dried over anhydrous magnesium sulfate, and after the desiccant was removed by filtration, the solvent was distilled off, and the residue was purified by silica gel column chromatography. eluted with 5% v / v methanol in methylene chloride to give 0.26 g of the title compound.

20

Nuclear Magnetic Resonance Spectrum [(CD3) 2SO] δ ppm 4.57 [1H, doublet, J = 5.0 Hz]; 4.90 [1H, singlet]; 5.27 [1H, doublet, J = 2.2 Hz]; 5.60 [1H, doublet of doublets, J = 2.2 & 5.0 Hz]; 6.21 [1H, singlet]; 6.70, 6.76 [total 1H, each singlet]; 7.2-7.6 [21H, multiplet]; Δ 8.95 [1H, singlet].

EXAMPLE 13 35 Dibenzhydryl-2-mercaptogriseolate 88 90427 with 0.25 g dibenzhydryl-1'-deadenino-1 '- [5 - amino-4- (N2-hydroxyamidino) imidazol-1-yl] griseolate (prepared as described in Example 12) was dissolved in a mixture of 2 ml of methanol, 2 ml of pyridine and 1 ml of carbon disulfide. The mixture was allowed to react at 80 ° C in 5 steel cylinders for 14.5 hours. The solvent was then removed by distillation. Toluene was added to the residue, and this procedure was repeated. The residue was purified by silica gel column chromatography eluting with 4% v / v methanol in methylene chloride to give 0.062 g of the title compound.

10

Nuclear Magnetic Resonance Spectrum [(CD3) 2SO] δ ppm 4.63 [1H, doublet, J = 5.0 Hz]; 4.88 [1H, singlet]; 5.26 [1H, doublet, J = 2.2 Hz]; 6.03 [1H, doublet of doublets, J = 2.2 & 5.0 Hz]; 6.39 [1H, singlet]; 6.69, 6.74 [total 1H, each singlet]; 6.9-7.5 [20H, multiplet]; 8.24 [1H, singlet].

EXAMPLE 14

Dimethyl-1'-deadenino-11B- (2-imino-f 1,2,4-oxadiazolo-f-3-hourin-7-25 v / v) griseolate hydrobromide (starting material of formula XXXIX which can be used to prepare compound (XL)) 8.46 g of dimethyl griseolate β-oxide (prepared as shown in Example 5-30) was suspended in 400 ml of methanol. To this was added 2.52 g of cyanogen bromide, and the mixture was stirred at room temperature for 1.5 hours, after which 630 mg of cyanogen bromide was added again, and it was further stirred for one hour. The solvent was removed by distillation under reduced pressure so that the volume of the remaining solution was about 100 ml. To this was added 100 ml of ethyl acetate, and distillation under reduced pressure was continued until the volume of the liquid was 1000 ml. To the remaining solution was gradually added 100 ml of ethyl acetate, stirring the mixture all the time. The suspension containing crystals was placed in the refrigerator overnight. The crystals were separated by filtration to give 9.77 g of the title compound.

Nuclear Magnetic Resonance Spectrum [(CD3) 2SO] δ ppm 3.67 [3H, singlet]; 3.73 [3H, singlet]; 4.67 [1H, doublet, J = 5.0 Hz]; 4.66 [1H, singlet]; 5.26 [1H, doublet, J = 2.2 Hz]; 5.84 [1H, doublet of doublets, J = 2.2 & 5.0 Hz]; Δ 6.76 [1H, singlet]; 9.07 [1H, singlet]; 10.23 [1H, singlet].

20 EXAMPLE 15

N1-oxide of dimethyl N6-svanogriseolate (starting material of formula XXXIX which can be used to prepare compound (XL)) 9.77 g of dimethyl-1'-deadenine-1 '/ 9- (2-imino- [1 The 2,4-oxadiazolo [3,2-i] -purin-7-yl) griseolate hydrobromide (prepared as described in Example 14) was dissolved in 50 ml of methanol. To this mixture was added 50 ml of a 20% v / v methanol solution of ammonia, after which the mixture was allowed to stand at room temperature for 60 minutes. To the reaction mixture was added 200 ml of ethyl acetate, and the solvent was removed by evaporation under reduced pressure using a suction device. After the volume of the liquid was 100 ml, 200 ml of ethyl acetate was added thereto, which was then removed by distillation under reduced pressure. After a liquid volume of 200 ml was obtained, the resulting mixture was placed in a refrigerator overnight.

90 90427

The crystals thus formed in the solution were filtered to give 8.53 g of the title compound as pale yellow fine crystals.

Nuclear Magnetic Resonance Spectrum [(CD3) 2SO] δ ppm δ 3.67 [3H, singlet]; 3.74 [3H, singlet]; 4.63 [1H, doublet, J = 5.0 Hz]; 4.66 [1H, singlet]; 5.16 [1H, doublet, J = 2.2 Hz]; 5.93 [1H, doublet of doublets, J = 2.2 & 5.0 Hz]; 6.50 [1H, singlet]; 8.33 [1H, singlet]; 8.49 [1H, singlet], 15. EXAMPLE 16

Dimethyl N-benzylloxy-N-cyanogriseolate 20 (- compound of formula (XL) according to the invention) 8.53 g of dimethyl N6-cyanogriseolate H1-olcsIdLa. (prepared as described in Example 15) was dissolved in 100 ml of dimethylformamide. To this solution were added 10 ml of benzyl bromide and 10 ml of triethyl-25 amine, and the mixture was stirred at room temperature for 4 hours. The solvent was distilled off under reduced pressure. To the residue were added 30 ml of ethanol and 30 ml of toluene, and then removed by distillation. This procedure was repeated 4 times. The oily substance thus obtained was mixed with 500 ml of diethyl ester, sonicated, stirring constantly with 30 spatulas, to give a pale yellow powder. This material was filtered off and dissolved in 300 ml of water and 500 ml of ethyl acetate. The separated organic layer was washed with 100 ml each of saturated aqueous sodium chloride solution, 0.2 N aqueous hydrochloric acid solution, 10% w / v aqueous sodium bicarbonate solution and saturated aqueous sodium chloride solution, respectively, and dried over anhydrous magnesium sulfate 90427. After the solution was treated with activated carbon, the solvent was removed by evaporation under reduced pressure to give 8.8 g of the title compound as a yellow caramel-like substance.

5

Nuclear Magnetic Resonance Spectrum [(CD3) 2SO] δ ppm 3.68 [3H, singlet]; 3.76 [3H, singlet]; 4.61 [1H, doublet, J = 5.0 Hz]; 4.67 [1H, singlet]; 5.26 [1H, doublet, J = 2.2 Hz]; 5.37 [2H, singlet]; 5.83 [1H, doublet of doublets, J = 2.2 & 5.0 Hz]; 6.57 [1H, singlet]; 7.2-7.8 [5H, multiplet]; 8.56 [1H, singlet]; 8.81 [1H, singlet].

20 EXAMPLE 17

Dimethyl N6-svano-N1-methoxygriseolate (- compound of formula (XL) according to the invention) 8.3 ml of triethylamine and 3.9 ml of methyl iodide were added to 40 ml of dimethylformamide containing 4.5 g of dimethyl-N6-cyano -griseolate β-oxide (prepared as described in Example 15) and kept cold by ice. The mixture was stirred at room temperature for 4.5 hours. The solvent was then removed by distillation, and 100 ml of diethyl ether was added to the residue, followed by obtaining a powder when the mixture was decomposed with a spatula. The insoluble material was filtered off, washed with 30 ml of diethyl ether and dissolved in a mixture of 80 ml of ethyl acetate and 20 ml of water, after which the solution was extracted repeatedly with ethyl acetate. The ethyl acetate layer was dried over anhydrous magnesium sulfate. After the drying agent was separated by filtration, the solvent was distilled off to give 3 g of the title compound.

Nuclear Magnetic Resonance Spectrum [(CD3) 2SO] δ ppm δ 3.60, 3.73 [total 3H, each singlet]; 4, IA [3H, singlet]; 4.58 [1H, doublet, J = 5.0 Hz); 4.64 [1H, singlet); 5.21 [1H, doublet, J = 2.2 Hz); 5.83 [1H, doublet of doublets, J = 2.2 & 5.0 Hz); 6.55 [1H, singlet); 8.53, 8.93 [total 1H, each singlet).

EXAMPLE 18 Preparation of a Drug from a Compound of Formula (XL).

20 2-Amino-N6-benzyloxypresolic acid 100 ml of a 0.2 N aqueous solution of sodium hydroxide was added to 400 ml of methanol containing 8.8 g of dimethyl N1-benzyloxy-N6-cyano-griseolate (prepared as described in Example 16). ), and which was stirred throughout. The pH of the reaction solution was adjusted to 12-12.5 by adding 1N aqueous sodium hydroxide solution, followed by standing at room temperature for 60 minutes. The solvent was distilled off to give a volume of about 100 ml of residual liquid. The resulting solution was mixed with 50 ml of water and 150 with 30 ml of ethanol, and heated under reflux for 2 hours. The solvent was then removed again by distillation under reduced pressure until the volume of the liquid was reduced to about 100 milliliters. To this was added 100 ml of 2N aqueous sodium hydroxide solution, and the resulting solution was allowed to stand at room temperature for 60 minutes. To the solution was added 200 ml of ethyl acetate, and the pH was adjusted to 0.5 with concentrated hydrochloric acid while stirring the solution. The aqueous layer and the 93 9 0 4 2 7 organic layer were separated, and the organic layer was washed with 500 ml of a 0.1 N aqueous hydrochloric acid solution, which was combined with the aqueous layer and treated with activated carbon. The pH of the solution was adjusted to 2.3, followed by the addition of solid sodium bicarbonate with vigorous stirring. The insoluble precipitate formed in the solution was placed in the refrigerator overnight. The resulting powder was filtered and dried to give 5.29 g of the title compound as a pale yellow powder.

Nuclear Magnetic Resonance Spectrum [(CD3) 2SO] δ ppm 4.54 [1H, singlet]; 4.55 [1H, doublet, J = 5.0 Hz]; 5.06 [2H, singlet]; 5.08 [1H, doublet, J = 2.2 Hz]; 5.83 [1H, doublet of doublets, J = 2.2 & 5.0 Hz]; 6.27 [1H, singlet]; 7.2-7.6 [5H, multiplet]; 7.79 [1H, singlet], 20. EXAMPLE 19 Preparation of a drug from a compound of formula (XL).

25 2-amino-N6-methoxygriseolinhaQQ

50 ml of a 0.2 N aqueous solution of sodium hydroxide was added to 70 ml of methanol containing 3 g of dimethyl N6-cyano-N1-methoxygriseolate (prepared as described in Example 17). The resulting mixture was stirred at room temperature for 1.5 hours, after which it was adjusted to pH 11.7 with 2 ml of 1 N aqueous sodium hydroxide solution, and the mixture was further stirred for 30 minutes. The pH of the solution was adjusted to 7.0 with concentrated hydrochloric acid and the methanol was removed by distillation. The remaining solution was stirred in 70 ml of ethanol and heated at reflux for 1.5 hours. The solvent was removed by evaporation to leave about 50 ml of solution, and the resulting solution 94 90427 was stirred in 50 ml of 1 N aqueous sodium hydroxide solution and stirred at room temperature for 60 minutes. The pH of the resulting mixture was adjusted to 1 with concentrated hydrochloric acid, and then washed with ethyl acetate. The pH of the aqueous layer was adjusted to 2.3 with 10% w / v aqueous sodium bicarbonate solution. It was then purified by column chromatography using a prepacked RP-8 column (reverse phase type, Merck) eluted with an aqueous solution containing 5% v / v acetonitrile and 0.02% v / v acetic acid to give 1.91 g of the title compound. compound.

10

Nuclear Magnetic Resonance Spectrum [(CD3) 2SO] δ ppm 3.77 [3H, singlet]; 4.51 [1H, singlet]; 4.54 [1H, doublet, J = 5.0 Hz]; 5.07 [1H, doublet, J = 2.2 Hz]; 5.87 [1H, doublet of doublets, J = 2.2 & 5.0 Hz]; 6.26 [1H, singlet]; 7.77 [1H, singlet].

EXAMPLE 20 (Preparation of a drug from the compound of Example 16) .V 25: 2-Amino-N6-hydroxygriseolinic acid; - ·] 200 mg of 2-amino-N6-benzyloxygriseolinic acid (prepared as described in Example .18) was heated in a mixture of 20 ml. methanol and 30 to 20 ml of water. The solution was then allowed to stand at room temperature under ice-cooling. To this was added 50 mg of carbon-bound palladium (10% w / w), and the mixture was stirred under hydrogen. As the reaction solution became cloudy in about 30 minutes due to the formation of a white substance *, it was clarified with 5 ml of a 1 N aqueous hydrochloric acid solution, and the resulting solution was stirred for 1.5 hours. The catalyst was removed by filtration, the methanol was removed by distillation under reduced pressure, and the pH of the solution was adjusted to 2.2 by adding saturated aqueous sodium bicarbonate solution with stirring. The reaction mixture was allowed to stand in an ice bath for 30 minutes, and the resulting yellowish solid was separated by filtration to give 5,100 mg of the title compound.

Nuclear Magnetic Resonance Spectrum [(CD3) 2SO] δ ppm 4.54 [1H, singlet]; 4.57 [1H, doublet, J = 5.0 Hz]; 5.11 [1H, doublet, J = 2.2 Hz]; 5.95 [1H, doublet of doublets, J = 2.2 & 5.0 Hz]; 6.31 [1H, singlet]; 7.83 [1H, singlet].

EXAMPLE 21 Preparation of a Drug from a Compound of Formula (XL).

20

Dimethyl 2-amino-N6-benzyloxygriseolate 5.4 g of dimethyl N1-benzyloxy-N6-cyanogriseolate (prepared as described in Example 16) was dissolved in 100 ml of methanol. To this was added 100 ml of 0.25 M phosphate buffer solution, and the mixture was heated under reflux for 4 hours. Crystals formed when methanol was distilled off. After the methanol was almost distilled, the pH of the solution was adjusted to 9 with 10% w / v aqueous sodium bicarbonate solution. The mixture was sonicated for 15-20 minutes, and the resulting crystals were filtered and dried to give 2.14 g of the title compound. The mother liquor was then adjusted to pH 11 with 2N aqueous sodium hydroxide solution and allowed to stand at room temperature overnight. The pH of the resulting solution was adjusted to 0.1 with concentrated hydrochloric acid, and the solution was treated with activated carbon, adjusted to pH 2.3 with 2N aqueous sodium hydroxide solution, and allowed to stand at 96 90427 at 5 ° C overnight. The precipitated material was filtered off to give 2.0 g of the title compound of Example 18.

Nuclear Magnetic Resonance Spectrum [(CD3) 2SO] δ ppm δ 3.63, 3.69 [total 3H, each singlet]; 4.51 [1H, doublet, J = 5.0 Hz]; 4.62 [1H, singlet]; 5.03 [2H, singlet]; 5.09 [1H, doublet, J = 2.2 Hz]; 5.83 [1H, doublet of doublets, J = 2.2 & 5.0 Hz]; 6.23 [1H, singlet]; 7.2-7.6 [5H, multiplet]; 7.70 [1H, singlet], 15 EXAMPLE 22 (Preparation of a drug from the compound of Example 16) 2 2-Aminopriseolinic acid 100 mg of 2-amino-β-benzyloxygriseic acid (prepared as described in Example 18) was dissolved in a mixture of 20 ml of a 1 N solution of hydrochloric acid. and 20 ml of acetone. To the solution was added 1 ml of Raney nickel (W-2), and the mixture was stirred at room temperature for 2.5 hours. Raney nickel was filtered off and the pH of the reaction mixture was adjusted to 2.3. It was purified by column chromatography using a prepacked RP-8 column (Merck) eluted with 3% v / v aqueous acetonitrile. The actual fraction was lyophilized to give 12 g of the title compound as a white powder.

Nuclear Magnetic Resonance Spectrum [(CD3) 2SO] δ ppm 35 4.46 [1H, singlet]; 4.55 [1H, doublet, J = 5.0 Hz]; 97 9 0 4 2 7 5.02 [1H, doublet, J = 2.2 Hz]; 5.87 [1H, doublet of doublets, J = 2.2 & 5.0 Hz]; 6.28 [1H, singlet]; 7.90 [1H, singlet].

5

Thin layer chromatography (ratio of the Rf value of the obtained compound to the Rf value of griseolinic acid, which was set to 1.0).

On silica gel plate (Merck): 0.80 (developing solution: water: methanol: aceto-10 nitrile 70:15:15 by volume) Reverse phase coated RP-8 plate: 0.79 (developing solution: water containing 2% v / v acetonitrile and 0.02% v / v acetic acid).

EXAMPLE 23 (Preparation of a drug from the compound of Example 16)

Dibenzhydryl-amino-N, N-benzyloxypriseolate 1.0 g of 2-amino-N6-benzyloxygriseolinic acid (prepared as described in Example 18) was suspended in 100 ml of acetone and 100 ml of water. Diphenyldiazomethane was added until the disappearance of its red color could no longer be observed. The reaction mixture was then stirred while adding 4 ml of a 1N aqueous hydrochloric acid solution; and diphenyldiazomethane was added again until the disappearance of its red color could no longer be observed. The mixture was then stirred for 60 minutes. The acetone was removed by distillation under reduced pressure, and water was removed by decantation. The residue was dissolved in a mixture of 50 ml of ethyl acetate and 50 ml of water, and the organic layer was washed with 30 ml of 5% w / v aqueous sodium bicarbonate solution. . and 30 ml of a saturated aqueous solution of sodium chloride, followed by drying over anhydrous magnesium sulfate. The solvent was removed by distillation under reduced pressure, and the residue was dissolved in 30 ml of 35 ethyl acetate, and the resulting solution was poured into 500 ml of hexane with stirring. The resulting insoluble material 98 90427 was collected by filtration and purified by chromatography using a prepacked silica gel column (Merck) eluted with methylene chloride containing 2.5% v / v methanol. From the two actual fractions, the one that eluted later was collected, evaporated to dryness and lyophilized from benzene to give 430 g of the title compound as a white powder.

Nuclear Magnetic Resonance Spectrum [(CD3) 2SO] δ ppm δ 4.63 [1H, doublet, J = 5.0 Hz]; 4.99 [1H, singlet]; 5.07 [2H, singlet]; 5.31 [1H, doublet, J = 2.2 Hz]; 5.97 [1H, doublet of doublets, J = 2.2 & 5.0 Hz]; 6.33 [1H, singlet]; 6.75 [1H, singlet]; 6.81 [1H, singlet]; 7.74 [1H, singlet].

20 EXAMPLE 24

Benzylhydryl 2-amino-N6-benzyloxy-N6-benzhydrylligriseolate The reaction mixture was purified by chromatography on a silica gel column following the procedure described in Example 23. The first actual fraction was collected, evaporated to dryness and lyophilized from Bentene to give 420 mg of the title compound.

Nuclear Magnetic Resonance Spectrum [(CD3) 2SO] δ ppm 4.53 [2H, broad singlet]; 4.72 [1H, doublet, J = 5.0 Hz]; 4.96 [1H, singlet]; 5.31 [1H, doublet, J = 2.2 Hz]; 6.10 [1H, doublet, J = 2.2 & 5.0 Hz]; 99 90427 6.46 [1H, singlet]; 6.75 [1H, singlet]; 6.82 [1H, singlet]; 7.69 [1H, singlet]; Δ 8.13 [1H, singlet].

EXAMPLE 25 Dibenzhydryl-2-aminogriseolate 83 mg of dibenzhydryl-2-amino-N6-benzyloxygriseolate (prepared as described in Example 23) was dissolved in a mixture of 20 ml of acetone and 10 ml of a 1N aqueous hydrochloric acid solution. To the mixture was added 1 15 ml of Raney nickel (W-2), after which the mixture was stirred vigorously at room temperature for 45 minutes. The Raney nickel was removed by filtration, and the solvent was removed by distillation under reduced pressure, continuing the distillation until the odor of acetone could no longer be detected. The resulting mixture was stirred in 30 ml of ethyl acetate and separated. The organic layer was washed with 20 ml of a 10% w / v aqueous solution of sodium bicarbonate and 20 ml of a saturated aqueous solution of sodium chloride, and dried over anhydrous magnesium sulfate. The solvent was removed by distillation under reduced pressure, and the residue was purified by chromatography on a prepacked silica gel column (Merck), eluting with 5% v / v methanol in methylene chloride. The actual fraction was collected and lyophilized from benzene to give 73 mg of the title compound as a white powder.

Nuclear Magnetic Resonance Spectrum [(CD 3) 2 SO] δ ppm 4.67 [1H, doublet, J = 5.0 Hz]; 4.92 [1H, singlet]; 5.27 [1H, doublet, J = 2.2 Hz]; 6.03 [1H, doublet of doublets, J = 2.2 & 5.0 Hz]; 35 6.37 [1H, singlet]; 6.73 [1H, singlet]; 100 90427 6.77 [1H, singlet]; 7.97 [1H, singlet], 5 EXAMPLE 26 0.56 g of dibenzhydryl-2-aminogriseolate (prepared as described in Example 25) was suspended in 5 ml of anisole and solubilized by adding 5 ml of trifluoroacetic acid suspension under ice-cold, after which the mixture was allowed to stand at room temperature. at a temperature of 10-15 minutes. To the reaction mixture was added 15 ml of toluene, and the solvent was then removed by distillation. The procedure in which a solvent mixture of 5 ml of acetone and 15 ml of toluene was added to the mixture, and this solvent mixture was then removed by distillation, was repeated twice, and the residue was suspended in 2 ml of acetone. The resulting suspension was poured into 200 mL of hexane with constant stirring. The resulting powdery material was filtered off and dissolved in 10% aqueous sodium bicarbonate. The pH of the resulting solution was adjusted to 0.6 with concentrated hydrochloric acid, and the solution was treated with activated carbon. The pH of the resulting mixture was adjusted to 2 with 10% aqueous sodium bicarbonate and allowed to stand at 5 ° C overnight. The separated crystals were separated by filtration and dried to give 0.15 g of the title compound. The mother liquor was purified by chromatography using 25 reverse phase packed columns packed with RP-8 phase (Merck) eluting with 3% v / v aqueous acetonitrile to give 0.04 g of the title compound having the same properties as in Example 1. With 22 products.

30 EXAMPLE 27

Dibenzhydryl-2-hydroxypriseolate 0.6 g of dibenzhydryl-2-aminogriseolate (prepared as described in Example 25) was dissolved in 50 ml of an 80% v / v aqueous solution of acetic acid.

The air in the vessel was displaced with nitrogen, and then 1 g of sodium nitrite was added to the mixture ιοί 90427 while keeping the mixture cold on ice, and the mixture was allowed to react at room temperature for 1.5 hours. The solvent was removed by distillation, and the residue was mixed with water, and water was similarly removed by distillation. The separated material was suspended in water and the water was removed by the same distillation. The separated material was suspended in water and collected by filtration. The residue was dissolved in 15 ml of acetone, and the pH of the solution was adjusted to 9-10 by adding concentrated aqueous ammonia solution. The resulting solution was allowed to stand at room temperature for 20 minutes, after which the acetone was removed by distillation. The residue was mixed with 50 ml of ethyl acetate and 50 ml of water and stirred thoroughly. The resulting precipitate was filtered off and purified by silica gel column chromatography (eluted with 10% v / v methanol in methylene chloride) to give 0.35 g of the title compound.

The separated ethyl acetate layer was washed with 10% w / v aqueous sodium bicarbonate solution and saturated aqueous sodium chloride solution, and dried over anhydrous magnesium sulfate. The desiccant was removed by filtration and the solvent was distilled off. The resulting residue was purified by silica gel column chromatography using 10% v / v methanol in methylene chloride as the eluent to give 0.04 g of the title compound.

Nuclear Magnetic Resonance Spectrum ((CD3) 2SO] δ ppm 4.59 [1H, doublet, J = 5.0 Hz], 4.90 [1H, singlet], 5.25 [1H, doublet, J = 2.2 Hz]; 6.05 [1H, doublet of doublets, J = 2.2 & 5.0 Hz]; 6.30 [1H, singlet]; 6.67, 6.73 [total 1H, each singlet]; .0-7.5 [20H, multiplet]; 7.93 [1H, singlet], 35 102 90427

F.STMKRKKT? R

Dibenzhydrol-6-desamino-2,6-dihydroxygriseolate In the procedure described in Example 27, the precipitated material separated by filtration from a mixture of ethyl acetate and water was chromatographed on a silica gel column. First, the title compound of Example 27 was eluted with methylene chloride containing 10% v / v methanol; the next fraction was eluted with methanol to give 0.06 g of the title compound 10.

Nuclear Magnetic Resonance Spectrum [(CD3) 2SO] δ ppm 4.47 [1H, doublet, J = 5.0 Hz); 4.85 [1H, singlet]; 5.19 [1H, doublet, J = 2.2 Hz]; 6.04 [1H, doublet of doublets, J = 5.0 & 2.2 Hz); 6.22 [1H, singlet]; 6.65, 6.73 [total 1H, each singlet]; 20.15-7.45 [20H, multiplet]; 7.56 [1H, singlet].

EXAMPLE 29 2-Hydroxygriseolinic acid 0.37 g of dibenzhydryl 2-hydroxygriseolate (prepared as described in Example 27) was dissolved in 3 ml of anisole. To this was added 3 ml of a solution of trifluoroacetic acid on ice while keeping cold, and the mixture was allowed to stand at room temperature for 30 minutes. Toluene was then added to the mixture, after which the solvent was removed by distillation. A mixture of acetone and toluene was added to the residue, which was then removed by distillation. The procedure was repeated twice and the resulting mixture was suspended in 2 ml of acetone and poured into 100 ml of hexane: 35 with constant stirring. The separated material was collected by filtration. The residue was dissolved in 10% w / v aqueous sodium bicarbonate solution, and the pH of the 103 90427 solution was adjusted to 1.2 with concentrated hydrochloric acid. The resulting mixture was purified by column chromatography using a prepacked RP-8 column (Merck) eluting with water to give 0.17 g of the title compound.

5

Nuclear Magnetic Resonance Spectrum [(CD3) 2SO] δ ppm 4.46 [1H, singlet]; 4.47 [1H, doublet, J = 5.0 Hz]; 5.05 [1H, doublet, J = 2.2 Hz]; 5.89 [1H, doublet of doublets, J = 2.2 & 5.0 Hz]; 6.24 [1H, singlet]; 7.95 [1H, singlet].

Thin layer chromatography (ratio of the Rf value of the obtained compound to the Rf value of griseolinic acid set to 1.0) Reverse phase RP-8 coated plate (Merck): 1.32 (developing solution: water containing 2% v / v acetonitrile and 0.02% v / v acetic acid).

EXAMPLE 30 6-Desamino-2,6-dihydroxygriseolinic acid 25 40 mg of dibenzhydryl-6-desamino-2,6-dihydroxygriseolinic acid (prepared as described in Example 28) was dissolved in 1 ml of anisole. To this solution was added 1 ml of a solution of trifluoroacetic acid on ice while keeping cold, and the mixture was allowed to stand at room temperature for 10-15 minutes. The procedure in which toluene was added and removed by distillation, after which a mixture of acetone and toluene removed by distillation was added to the mixture was repeated twice, and the resulting residue was suspended in 0.5 ml of acetone. After 20 ml of hexane was added to the suspension, the separated material was collected by filtration. The residue was dissolved in a 10% w / v aqueous solution of sodium bicarbonate. The pH of the solution was adjusted to 0.6 with concentrated hydrochloric acid. The resulting solution was purified by column chromatography using a 104 9 0 427 prepacked RP-8 column (reverse phase type, Merck) eluting with water to give 25 mg of the title compound.

Nuclear Magnetic Resonance Spectrum [(CD3) 2SO] δ ppm δ 4.49 [1H, singlet]; 4.58 [1H, doublet, J = 5.0 Hz]; 5.12 [1H, doublet, J = 2.2 Hz]; 5.53 [1H, doublet of doublets, J = 2.2 & 5.0 Hz]; 6.41 [1H, singlet]; 7.91 [1H, singlet].

Thin layer chromatography (ratio of the Rf value of the obtained compound to the Rf value of griseolinic acid, which was set to 1.0) 15

Silica gel coated plate (Merck): 0.57 (developing solution: water: methanol: acetonitrile 70:15:15 by volume); Reverse Phase RP-8 Coated Plate (Merck): 1.65 (developing solution: water, 20 containing 2% v / v acetonitrile and 0.02% v / v acetic acid).

EXAMPLE 31

Dimethyl N6-benzyloxy-2- (N, N'-dimethylaminomethylene-aminogriseo-25) 528 mg of dimethyl-2-amino-.alpha.-benzyloxygriseolate (prepared as described in Example 21) was dissolved in 10 ml of dimethylformamide. 24 ml of dimethylformamide dimethylacetal and the mixture was allowed to stand at room temperature for 2 hours, after the disappearance of the starting material was confirmed by thin layer chromatography, the solvent was removed by filtration under reduced pressure, the residue was dissolved in 40 ml of methylene chloride and 40 ml of water. 20 ml of a saturated aqueous solution of sodium chloride 35. All the aqueous layers obtained were combined and then mixed with 2 ml of a 5% w / v 105% aqueous solution of sodium bicarbonate and extracted twice with 20 ml of methylene chloride The organic layers were combined, dried over anhydrous magnesium sulfate and the solvent removed. by distillation under reduced pressure. The residue was chromatographed on a silica gel column (Merck) eluting with 3% v / v methanol in methylene chloride. After purification, the obtained substance was lyophilized from benzene to give 322 mg of the title compound as a white powder.

Nuclear Magnetic Resonance Spectrum [(CD3) 2SO] δ ppm δ 3.04 [3H, singlet]; 3.19 [3H, singlet]; 3.67 [3H, singlet]; 3.72 [3H, singlet]; 4.47 [1H, doublet, J = 5.0 Hz]; 4.63 [1H, singlet]; 5.01 [2H, singlet]; 5.12 [1H, doublet, J = 2.2 Hz]; 6.00 [1H, doublet of doublets, J = 2.2 & 5.0 Hz]; Δ 6.38 [1H, singlet]; 7.2-7.6 [5H, multiplet]; 7.83 [1H, singlet]; 8.50 [1H, singlet].

25 EXAMPLE 32

Dimethyl 2- (N'-N'-dimethylaminomethyleneaminoprisezolate) 3083 mg of dimethyl-N6-benzyloxy-2- (N ', N'-dimethylaminomethylene) aminogreizate (prepared as described in Example 31) was dissolved in 100 ml of acetone and 75 ml of 1 N To this was added 10 ml of Raney nickel (W-2) suspended in water, and the resulting mixture was stirred at room temperature for 30 minutes, maintaining a pH of at least 1.0, followed by a pH meter, the Raney nickel was removed by filtration, and the filtrate was concentrated. After almost complete removal of acetone, the resulting mixture was stirred in 200 ml of methylene chloride and neutralized with aqueous sodium bicarbonate solution, and the resulting insoluble material was removed by filtration After separating the organic layer by 50 ml, the aqueous layer was extracted three times. methylene chloride the mixtures were combined and dried over anhydrous magnesium sulfate. The solvent was removed by distillation under reduced pressure, and the residue was chromatographed on a silica gel column (Merck) eluting with 5% 10% v / v methanol in methylene chloride. The resulting material was purified and lyophilized from benzene to give 180 mg of the title compound as a white powder.

Nuclear Magnetic Resonance Spectrum [(CD3) 2SO] δ ppm 3.04 [3H, singlet]; 3.15 [3H, singlet]; 3.67 [3H, singlet]; 3.73 [3H, singlet]; 4.53 [1H, doublet, J = 5.0 Hz]; 4.64 [1H, singlet]; 5.13 [1H, doublet, J = 2.2 Hz]; 6.06 [1H, doublet of doublets, J = 2.2 & 5.0 Hz]; 6.47 [1H, singlet]; 25 7.33 [1H, singlet], EXAMPLE 33

Dimethyl 6-desamino-2- (N * .N * -dimethylaminomethylene) -amino-6-hydroxy-30 eisolate 477 mg of dimethyl 2- (N'-dimethylaminomethylene) aminogriseolate (prepared as described in Example 32) was dissolved in 50 ml of acetic acid 80 % v / v aqueous solution. To this was added 1.34 g of a solution of sodium nitrite-35 on ice while keeping the mixture cold, after which the mixture was allowed to stand at room temperature for 17 hours. After the disappearance of the starting material was confirmed by thin layer chromatography, the solvent was removed by distillation under reduced pressure. Ethanol was added to the residue, which was then removed by distillation, and this addition of ethanol and removal by distillation was repeated so many times that the odor of acetic acid 5 could no longer be detected. The residue was dissolved in a mixture of 50 ml of methylene chloride, 20 ml of water and 5 ml of 5% w / v aqueous sodium bicarbonate solution. The organic layer was separated and extracted three times with 30 ml of methylene chloride each time, and the resulting extracts were combined. The solvent was removed by distillation under reduced pressure. Residue 10 was purified by chromatography on a prepacked silica gel column (Merck) eluting with 10% v / v methanol in methylene chloride. The actual fractions were collected and lyophilized from benzene to give 310 mg of the title compound as a white powder.

15

Ultraviolet absorption spectrum (methanol) λ max: acidic: 292 nm neutral: 300 nm 20 basic: 279 nm.

EXAMPLE 34 2-Amino-6-desamino-6-hydroxygriseoleic acid 130 mg of dimethyl 6-desamino-2- (N ', N'-dimethylaminomethylene) amino-6-hydroxygriseolate (prepared as described in Example 33) was dissolved in 20 ml of concentrated ammonia. aqueous solution, and the mixture was allowed to stand at room temperature for 3 hours. The solution was evaporated to dryness under reduced pressure, and the residue was dissolved in 10 ml of water. The pH of the resulting solution was adjusted to 2.3, after which it was chromatographed using a prepacked RP-8 chromatography column (Merck), which was washed with water and eluted with water containing 5% v / v acetonitrile. The actual fractions were collected and lyophilized to give 67 mg of the title compound as a white powder.

108 90 427

Ultraviolet absorption spectrum (H2) λ max: acidic: 225 nm, 273 nm ("shoulder") neutral: 253 nm, 278 nm ("shoulder") 5 basic: 264 nm.

Nuclear Magnetic Resonance Spectrum [(CD3) 2SO] δ ppm 4.48 [1H, singlet]; 4.53 [1H, doublet, J = 4.9 Hz]; 5.07 (1H, doublet, J = 2.4 Hz]; 5.79 [1H, doublet of doublets, J = 2.4 & 4.9 Hz]; 6.25 [1H, singlet]; 7.87 [ 1H, singlet], 15

Thin layer chromatography (ratio of the Rf value of the compound obtained to the Rf value of griseolinic acid, set to 1.0):

Silica gel coated plate (Merck): 0.80 (developing solution water: 20 methanol: acetonitrile 70:15:15, by volume); Reverse Phase RP-8 Coated Plate (Merck): 1.44 (developing solution: water containing 2% v / v acetonitrile and 0.02% v / v acetic acid).

. 25 EXAMPLE 35

Methyl 2-amino-6-desamino-6-hydroxyeriserate 30 418 mg of silver perchlorate was added to a suspension of 400 mg of 2-amino-6-desamino-6-hydroxygriseolinic acid (prepared as described in Example 34) in 40 ml of methanol, and to the resulting mixture 0.187 ml of methyl iodide was added, the mixture being stirred at room temperature. The mixture was stirred continuously at room temperature for a further 2.5 hours, after which 0.126 ml of methyl iodide was added again. The resulting mixture was further stirred for 8,109,90427 hours, after which it was placed in the refrigerator overnight. The separated insoluble material was separated by filtration, and methanol was removed by distillation under reduced pressure. The residue was dissolved in water, and the pH of the resulting solution was adjusted to 1.38 with concentrated aqueous hydrochloric acid, and the solution was purified by chromatography through a packed RP-8 column (Merck) using 5% v / v acetonitrile in water as eluent. The obtained main fraction was freeze-dried to give 170 mg of the title compound as a white powder.

10

Ultraviolet absorption spectrum (H 2 O) λ max: acidic: 257.5 nm, 280 nm ("shoulder") neutral: 253.5 nm, 275 nm ("shoulder") 15 basic: 265 nm.

Nuclear Magnetic Resonance Spectrum [(CD3) 2SO] δ ppm 4.53 [1H, doublet, J = 5.4 Hz]; Δ 4.59 [1H, singlet]; 5.09 [1H, doublet, J = 2.0 Hz]; 5.80 [1H, doublet of doublets, J = 2.0 & 5.4 Hz]; 6.25 [1H, singlet]; 7.88 [1H, singlet].

EXAMPLE 36 2-Amino-6-desamino-6-hydroxy-acioic acid monoamide 30

A solution of 120 mg of methyl 2-amino-6-desamino-6-hydroxy-griseolate (prepared as described in Example 35) in 20 ml of a 20% v / v methanol solution of ammonia was allowed to stand overnight. The solvent was then removed by distillation under reduced pressure, and the residue was dissolved in 3 ml of 1 N aqueous sodium hydroxide solution. The pH of the resulting solution was adjusted to 1.8-1.9 by adding a concentrated aqueous solution of chlorine 90427 hydrochloric acid, keeping it cold on ice, to form a gel-like material in situ. It was then dissolved in about 20 ml of water by adjusting the pH to 0.5. The pH of the resulting solution was then adjusted to 1.0 by the addition of aqueous sodium bicarbonate solution. The insoluble material was filtered off, and the residue was purified by column chromatography through a prepacked RP-8 column (Merck), using 5% v / v acetonitrile in water as the eluent. The actual fraction was lyophilized to give 79 mg of the title compound.

10

Ultraviolet Absorption Spectrum (H 2 O) Amaxnm (ε): acidic: 257 (12100), 280 ("shoulder") (8400); neutral: 252 (13700), 277 ("shoulder") (8800); 15 basic: 264 (12400).

Nuclear Magnetic Resonance Spectrum [(CD3) 2SO] δ ppm: 4.32 [1H, singlet]; 4.50 [1H, doublet, J = 4.9 Hz]; 5.05 [1H, doublet, J = 2.0 Hz]; 5.80 [1H, doublet of doublets, J = 2.4 & 4.9 Hz]; 6.24 [1H, singlet]; 7.78 [1H, singlet], 25 EXAMPLE 37 N, N-Methyl-4-deoxy-4 ', 5'-dihydropriseolinic acid 30 1 ml of methyl iodide was added to a solution of 100 mg of 7'-deoxy-4'ct, 5'- dihydrogriseolinic acid in 20 ml of dimethylformamide, and this mixture is allowed to stand at room temperature for 24 hours in a closed vessel. The solvent was removed by distillation under reduced pressure to give a residue. To the residue were added 10 ml of acetone and 10 ml of toluene, 35 and the mixture was concentrated by evaporation under reduced pressure. This procedure was repeated twice. A solution of the resulting residue 20 in 90427 ml of 0.5 M phosphate buffer, pH 7.0 was stirred at reflux for 3 hours to give a reaction mixture which was in turn purified by column chromatography using a pre-packed RP-8 column (Merck) followed by the major fraction was lyophilized to give 67 mg of the title compound as a white powder.

Ultraviolet Absorption Spectrum (ε) Amax: pH 1.0 262 nm (17700); 10 H 2 O 264 nm (16700); pH 13,266 nm (17100).

Nuclear Magnetic Resonance Spectrum [(CD3) 2SO + D2O] δ ppm: 2.28-2.31 [2H, multiplet]; 2.80-3.03 [5H, multiplet]; 4.37-4.46 [3H, multiplet]; 6.16 [1H, singlet]; 8.26 [1H doublet]; 8.28 [1H, singlet], PREPARATION 1

Dimethyl ligise 25 700 g of griseolinic acid were dissolved in 100 ml of dimethylformamide under ice-cooling. A solution of 1.0-1.2 millimoles of diazomethane in 1 ml of diethyl ether was added with stirring to this solution until the yellow color indicated the presence of diazomethane. The mixture was then allowed to stand for 10 minutes. Acetic acid was then added to the mixture to decompose excess diazomethane, which was then removed by evaporation under reduced pressure to give a residue. The residue was dissolved in methanol, and the insoluble material was separated by filtration. The filtrate was evaporated under reduced pressure to give a residue which was recrystallized from water to give 540 mg of the title compound.

112 90 42 7

Ultraviolet absorption spectrum (methanol) Amax nm: 258 (c - 15600).

Nuclear Magnetic Resonance Spectrum [(CD3) 2SO] δ ppm: 4.60 [1H, doublet, J = 6.0 Hz]; 4.66 [1H, singlet]; 5.12 [1H, doublet, J = 3.0 Hz]; 6.06 [1H, doublet of doublets, J = 3.0 & 6.0 Hz]; 6.53 [1H, singlet]; 8.33 [1H, singlet]; 8.37 [1H, singlet].

15 PREPARATION 2

Dlmetwli-02 '. O7'-diacetylligrole isolate 10 g of dimethyl griseolate (prepared as shown in Preparation 1) was dissolved in 150 ml of pyrimidine in a round-bottomed flask, and 33 ml of acetic anhydride was added under ice-cooling. The mixture was allowed to stand at room temperature for 2 hours. At the end of this time, 15 ml of water was added to the reaction mixture while keeping it cold, and the solvent was removed by evaporation under reduced pressure. The residue was dissolved in 400 ml of methylene chloride, and the resulting solution was washed with 400 ml of 1 N aqueous hydrochloric acid, 400 ml of water, and 400 ml of saturated aqueous sodium bicarbonate solution, respectively. The solution was then extracted twice with methylene chloride. The methylene chloride extracts were dried over anhydrous magnesium sulfate, and the solvent was removed by distillation under reduced pressure to give 6.70 g of the title compound as crystals.

Nuclear Magnetic Resonance Spectrum [(CD 3) 2 SO] δ ppm: 35 5.17 [1H, doublet, J = 3.0 Hz]; 5.66 [1H, doublet, J = 6.0 Hz]; 113 90 42 7 5.73 [1H, singlet]; 6.31 [1H, doublet of doublets, J = 3.0 & 6.0 Hz]; 6.89 [1H, singlet]; 8.23 [1H, singlet]; Δ 8.36 [1H, singlet], PREPARATION 3

Dimethyl O2 ', O7'-diacetyl-6-desamino-6-hydroxysgriseolate 2.55 g of sodium nitrite was added to a solution of 1.82 g of dimethyl O2', 07'-diacetylgriseolate (prepared as described in Preparation 2) in acetic acid. In an 80% v / v aqueous solution, the solution was kept cold on ice, and the mixture was allowed to stand for 16 hours in a tightly closed vessel. At this point, it was determined by thin layer chromatography that the starting material remained in the reaction mixture. A further 1 g of sodium nitrite was added to the mixture, and the mixture was allowed to stand for 3 hours. The solution was removed by evaporation and the resulting residue was dissolved in acetone. Toluene was added to the mixture, which was then removed by distillation. 20 This procedure was repeated three times.

The residue was dissolved in a mixture of water and chloroform. The organic layer was washed with aqueous sodium bicarbonate solution and saturated sodium chloride solution. . aqueous solution, and then dried over anhydrous magnesium sulfate. Evaporation of the solvent gave a light brown glass. This material was purified by silica gel column chromatography, followed by dissolving in a small amount of acetone. An appropriate amount of benzene was added to the solution, and the mixture was allowed to stand. The resulting white crystals were collected by filtration to give 1.28 g of the title compound as fine white crystals.

Nuclear Magnetic Resonance Spectrum [(CD3) 2SO] δ ppm: 5.22 [1H, doublet, J = 3.0 Hz]; 35.62 [1H, doublet, J = 6.0 Hz]; 5.73 [1H, singlet]; 114 90427 6.13 [1H, doublet of doublets, J = 3.0 & 6.0 Hz]; 6.88 [1H, singlet]; 8.18 [1H, singlet]; 8.34 [1H, singlet].

PREPARATION 4 6-Desamino-6-hydroxygriseolinic acid 10 A solution of 5.31 g of griseolinic acid in 80% v / v aqueous acetic acid was prepared by heating and then allowing to cool to room temperature. To the mixture was added 9.60 g of sodium nitrite under a nitrogen atmosphere. The mixture was allowed to stand for 16 hours in a tightly closed container. The solvent was removed from the mixture by evaporation under reduced pressure to give a residue. Ethanol was added to this residue, followed by distillation. This procedure was repeated so many times that the odor of acetic acid could no longer be detected in the mixture. The residue was dissolved in 50 ml of water and the pH was adjusted to 1.0 with concentrated hydrochloric acid, keeping it cold on ice. The mixture was allowed to stand in a refrigerator for 16 hours, and the resulting precipitate was collected by filtration, washed with a small amount of water, and recrystallized from a mixture of water and acetone to give 1.66 g of the title compound. Concentration of the mother liquor gave 2.20 g of crude crystals, which were also recrystallized to give a further 1.2 g of the title compound.

Nuclear Magnetic Resonance Spectrum [(CD3) 2SO] δ ppm: 4.50 [1H, singlet]; 4.57 [1H, doublet, J = 6.0 Hz]; 5.12 [1H, doublet, J = 3.0 Hz]; 5.88 [1H, doublet of doublets, J = 3.0 & 6.0 Hz]; 6.50 [1H, singlet]; 8.17 [1H, singlet]; 35 8.33 [1H, singlet].

115 90427 VATMTSTUS 5

Dimethyl 6-desamino-6-hydroxy-4,1'-chloro-5'-hydro-O2'.O7'-diacetylgrisolate 5 4 g dimethyl-O2 ', Q7'-diacetyl -6-desamino-6-hydroxygriseolate (prepared as described in Preparation 3) and 40 ml of acetic acid containing 4% w / v hydrogen chloride were placed in a two-necked flask equipped with a condenser under a nitrogen atmosphere. The mixture was heated at 80 ° C for 10 hours, after which the solvent was removed by evaporation under reduced pressure to give a residue. The residue was dissolved in toluene and methylene chloride, which were then removed under reduced pressure. This procedure was repeated three times. The residue was purified by silica gel column chromatography using methylene chloride containing 4% v / v methanol-15 as an eluent to give 2.0 g of the title compound.

Nuclear Magnetic Resonance Spectrum [(CD 3) 2 SO] δ ppm: 3.32 [1H, doublet, J = 15.0 Hz]; 3.75 [1H, doublet, J = 15.0 Hz]; 5.28 [1H, doublet, J = 4.5 Hz]; 6.00 [1H, singlet]; 6.28 [1H, doublet of doublets, J = 4.5 & 5.9 Hz]; 6.55 [1H, doublet, J = 5.9 Hz]; 8.18 (1H, singlet]; 8.47 [1H, singlet], PREPARATION 6 30

Dimethyl 6-desamino-6-hydroxy-O 2 ', 7'-diacetyl-4'-bromo-5'-hydro-griseolate

A mixture of 500 mg of dimethyl-O2 ', 07'-diacetyl-6-desamino-6 to 35 hydroxyglycolate (prepared as described in Preparation 3) and 10 ml of acetic acid containing 10% w / v hydrobromic acid was placed in a 116 90427 vessel, which was sealed, and the compound was dissolved by sonication for 30 minutes. The solution was then allowed to stand at room temperature for 64 hours. The solvent was removed by distillation under reduced pressure to give a residue to which acetone and toluene were added, which solvents were then removed by distillation. This procedure was repeated three times. The mixture comprising the resulting residue in 30 ml of ethyl acetate was sonicated, and filtered to recover insoluble material. This material was dissolved in a mixture of 30 ml of ethyl acetate and 30 ml of 5% w / v aqueous sodium bicarbonate solution and separated. The organic layer was washed with 20 ml of saturated aqueous sodium chloride solution and dried over anhydrous magnesium sulfate. The solution was evaporated under reduced pressure to give a residue which was in turn purified by chromatography on a column of silica gel using methylene chloride containing 3% v / v methanol as eluent. The solvent was removed from the actual fraction by evaporation under reduced pressure, and the residue was dissolved in benzene. The benzene solution was lyophilized to give 60 mg of the title compound as a white powder.

Nuclear Magnetic Resonance Spectrum [(CD3) 2SO] δ ppm: 2.98 [1H, doublet, J = 15.6 Hz]; 3.47 [1H, doublet, J = 15.6 Hz]; 5.35 [1H, doublet, J = 4.2 Hz]; 5.57 [1H, singlet]; 6.32 [1H, doublet of doublets, J = 6.6 & 4.2 Hz]; 6.53 [1H, doublet, J = 6.6 Hz]; 8.17 [1H, singlet]; 8.47 [1H, singlet], PREPARATION 7

Dimethyl-6-desamino-6-hydroxy-1-4,5'-dihydro-O2 ', O7'-diazephenyl-imiolate 35 117 90427 500 mg dimethyl-6-desamino-6-hydroxy-4') -chloro-5'-hydro-O2 ', 07'-diazyl styligrise (prepared as described in Preparation 5) and 10 mg of 2,2'-azobisisobutyronitrile were placed in a two-necked flask and dissolved in 20 ml of benzene under a nitrogen atmosphere. To this solution was added 3.1 ml of tributyl anhydride with a plunger, and the mixture was stirred at reflux for 2 hours. At the end of this time, the solvent was removed by evaporation under reduced pressure. The residue was dissolved in methylene chloride and purified by chromatography on a silica gel column eluting with 3% v / v methanol in methylene chloride. The actual fractions were concentrated by evaporation under reduced pressure to give 350 mg of the title compound.

Nuclear Magnetic Resonance Spectrum [(CDCl 3 + D 2 O] δ ppm: δ 2.40-2.70 [2H, multiplet]; 5.00 [1H, doublet, J = 4.5 Hz]; 5.62 [1H, singlet] ; 5.88 [1H, doublet of doublets; J = 4.5 & 7.5 Hz]; 6.48 [1H, doublet, J - 7.5 Hz]; 8.18 [1H, singlet]; 8.48 [1H, singlet].

PREPARATION 8 25 Dihydrodeoxygriseolinic acid

First, 30 liters of medium with a pH of 7.0 before sterilization and the following composition were prepared (percentages are expressed by weight per unit volume, w / v): 30 glucose 5% soy flour 1% yeast extract 0.1% polypeptone 0.4% 35 meat extract 0.4% sodium chloride 0.25% 118 90427 calcium carbonate 0.5% water, make up to 100%.

15 liters of this medium were each placed in two 30-liter tank-5 fermentors, which were then sterilized under pressure at 120 ° C for 30 minutes. The culture medium was cooled, after which 150 ml (1% by volume) of a culture of Streptomyces griseoaurantiacus strain SANK 63479 (previously prepared by incubating the strain in said medium, keeping 10 cultures in a rotary shaker at 28 ° C for 72 hours) were inoculated into each fermentor. The strain was cultured at 28 ° C for 96 hours with aeration at 15 liters per minute, and stirring at 200 rpm.

The two media were then filtered to remove mycelium, and the combined filtrate, with a total volume of 28 liters and a pH of 7.0, was passed through a Diaion HP 20 column (trademark for ion exchange resin from Mitsubishi Chemical Industries Ltd.) and then adsorbed onto a pad of activated carbon. column. This column was washed with water, and then the adsorbed material was eluted with a 60:40 (v / v) mixture of acetone and water. The acetone was evaporated from the resulting solution under reduced pressure, and the remaining aqueous solution was concentrated by evaporation under reduced pressure, and lyophilized to give 150 mg of a crude powder.

This crude powder was dissolved in a small amount of distilled water and then adsorbed onto Dowex 1x4 resin (in Cl 'form; trademark of the ion exchange resin produced by Dow Chemical Company). At this point, the product was a mixture comprising griseolinic acid and dihydrodesoxygriseolinic acid. This mixture was eluted using a gradient of sodium chloride to separate the two components, after which the eluate was subjected to column chromatography using a Sephadex LH-20 column (trademark of Pharmacia), and dihydro-deoxygriseolinic acid was eluted from this column with water. Fractions containing this material were combined and adjusted to pH 2.5 by the addition of 1N aqueous hydrochloric acid. The product was then adsorbed onto a Diaion HP 20 column, washed with water and then eluted with a mixture of acetone and water 119 90427 60:40 (v / v). The eluate was allowed to stand overnight at 4 ° C, at which time dihydrodeoxygriseolinic acid separated as a strip. This material was separated from the liquid to give a total of 1.87 mg of dihydrodesoxygriseolinic acid as white strips with a melting point of 160 ° C (decomposes at this temperature, turning brown). This compound was obtained as a single spot by thin layer chromatography on silica gel (silica gel part number 5715, a product of Merck & Co. Inc.).

10

Claims (3)

A compound of formula (XL): Si R 2 (X 2) where::: R 1 and R 2 are independently selected from a group comprising the hydrogen atom and the group of formula -OR 9; R3 and R4 are independently selected from a group comprising. . carbamoyl groups and carboxy groups and lower alkoxycarbonyl groups, and these lower alkoxycarbonyl groups are optionally substituted with a phenyl or benzhydryl group; R5 and R6 are both hydrogen atoms, or they form an additional carbon-carbon bond between the carbon atoms to which they are attached; R9 represents a hydrogen atom, a lower alkanoyl group, a benzyl group or a benzoyl group; and R 30 represents a lower alkyl group or aralkyl group; 123 90427 saint pharmaceutically acceptable salts and esters of these compounds. 2. Dimethyl-N1-benzyloxy-N6-cyanoglyseolate or pharmaceutically acceptable salts and esters thereof. 5 3. Dimethyl-N6-cyano-N1-methoxygrisolate or pharmaceutically acceptable salts and esters thereof.