FR2460953A1 - NOVEL PROCESS FOR THE PREPARATION OF ANOMERS AND ASTERIC MIXTURES OF NEW SPECTINOMYCIN ANALOGS - Google Patents

Abstract

THE INVENTION RELATES TO THE FIELD OF PHARMACEUTICAL CHEMISTRY. IT CONCERNS A NEW PROCESS FOR PREPARING NEW ANALOGUES OF SPECTINOMYCIN OF FORMULA: (CF DRAWING IN BOPI) IN THE FORM OF ANOMERAS AND ASTERIC MIXTURES.

Description

The present invention relates to anomers of certain novel analogues

  spectinomycin and their asteroid mixtures. The invention also relates to novel intermediate compounds for obtaining spectinomycin analogs. Processes for producing spectinomycin analogues are also within the scope

of the invention.

  Spectinomycin is a known antibiotic that responds to the following formula: OH

/ CH3 H

0CH3 "N 0t0 X CH3

N / \ H OH

  Until now, spectinomycin has been prepared by a microbiological process (see patent of

  United States of America N 3 234 092).

  Some spectinomycin analogs have been described by Rosenbrook Jr. et al. In "J. Antibiotics", 28, pages 953 and 960 (1975) and "J. Antibiotics", 31, page 451 (1978). In addition, Carney et al. Describe chlorodeoxy derivatives of

  spectinomycin in "J. Antibiotics", 30, 960 (1977). 9-

  epi-4 (R) -dihydrospectinomycin has further been described by Foley et al. in "J. Org. Chem.", 43, 22, pages

4355-4359 (1978).

  However, contrary to the present invention,

  No biological activity was reported for any of the analogues and derivatives of spectinomycin.

  described in the aforementioned references.

  The chemical processes of the prior art which are closest to the process of the invention teach a preferred reaction at the 5-hydroxyl group of the

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  2-deoxystreptamine (1) with tri-OO-acetyl-2 chloride

  deoxy-2-nitroso-α-D-glycopyranosyl (2) to form a

  pseudodisaccharide (3) according to the following scheme (see Lemieux, "Can J. Chem.", 51, page 53 (1973)): OH HCBz H

H +

2 ON Ac i

OH

OAc NH CBz (2)

(1)

  OH -N 0 X -OAc / NHO OAc-, HON, OAc OH H-NX OAc CBz (3)

  o CBz is a carbobenzyloxy group. Mallams and collabora-

  "J. Chem Soc." Perkin I, page 1118 (1976), extend the Lemieux reaction to the synthesis of disaccharides and trisaccharides. The elimination of oximes has been taught by Lemieux et al. In "Can. J. Chem." 51, page 19

  (1973) and by Mallams et al in "J. Chem Soc."

Perkin I, p. 1097 (1976).

  The compounds of the invention are anomers and asteroid mixtures of compounds which correspond to the formula:

3 2460953

R?

R B1 R6 R

Re _,

R10 R: 14 OHO

  Wherein R is a group of the formula:

R3 4

R4

  or R 1 to R 4 may be the same or different and are selected from hydrogen, lower alkyl, lower alkenyl, lower haloalkyl,

  amino-lower alkyl radical, an alkynyl radical

  a radical -OX and a radical - (CH2) n-OX, provided that R1 and R2 are not hydroxy radicals and that one of the symbols R3 and R4 represents a hydrogen atom, X is an atom of hydrogen or a lower alkyl, lower alkenyl or lower alkynyl radical, n is an integer having a value of 1 to 4, R5 is a hydrogen atom or a lower alkyl radical, the variables R6 to R14 are chosen between hydrogen, a lower alkyl radical, a lower alkenyl radical and a lower alkynyl radical, provided that

  one of the variables R7 and R8 is always a hydrogen atom

  gene and that one of the variables R11 and R12 is always a hydrogen atom, and B and B1 are equal or different and are selected from hydrogen and a radical hydroxy, alkoxy, o-lower alkenyl, thio, thio lower alkyl and lower thio-alkenyl; and A is an oxygen or sulfur atom; provided that when the compound meets the formula:

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OH

H H

CHN {0H CH

I OH

NCH3 -0

H

  B1 can not be a hydroxy group.

  The numbering of the carbon atoms indicated in the compound of formula I will be used in the comments

  concerning this compound in this specification.

  The compounds of the invention comprise the hydrated forms of the compounds of formula I. These compounds are hydrated at the 3 'position and correspond to the formula: R7

R - 2

B1 2

R10 OH

0N HO OH

R, R12 I

  in which A, B, B1, R and R6 to R14 have the definitions given above. The invention also relates to the pharmaceutically acceptable salts of the compounds of

formulas I and I '.

  In the designation of the variables of which it is

  question in this submission, the group "- (CH2) n"

  takes lower straight chain alkyl groups and their isomers. The term "lower alkyl" refers to methyl, ethyl, propyl, butyl, pentyl, hexyl,

  heptyl, octyl and their isomeric forms.

  "Lower alkenyl" expressioa refers to ethylidene, propylidene, butylidene, pentylidene, hexylidene, heptylidene, octylidene and their isomeric forms.

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  The term "lower alkynyl" refers to ethynyl, propynyl, butynyl, pentynyl, hexynyl,

  heptynyl, octynyl and their isomeric forms.

  The term "lower alkoxy" refers to methoxy, ethoxy, propoxy, butoxy, pentoxy, hexoxy,

  heptoxy, octoxy and their isomeric forms.

  The term "acyl" refers to formyl, acetyl, propionyl, butyryl and pentionyl groups and their isomeric forms. The term "aralkyl" refers to benzyl groups,

  phenethyl, phenpropyl, phenbutyl, phenpentyl, diphenyl-

  methyl, diphenyloctyl and their isomeric forms and the fluorenylmethyl group. The term "lower halogenalkyl" refers to a - (CH 2) n -halogeno group and its isomeric forms. This group

  contains one to three halo substituents.

  The term "amino-lower alkyl" refers to a group of the formula: lower alkyl (or H) (CH 2) n-N 1 (lower alkyl (or H)

and its isomeric forms.

  The term "aroyl" refers to the benzoyl group, a substituted benzoyl group, the naphthoyl group and a substituted naphthoyl group. The substituted benzoyl and naphthoyl groups may have one to three substituents selected from lower alkyl, lower alkoxy,

nitro and halo.

  The term "haloalkoxycarbonyl" refers to mono-, di-, tri-halomethoxycarbonyl; mono-,

  di-, tri-haloethoxycarbonyl; mono-, di-, tri-halogeno-

  propoxycarbonyl; mono-, di-, tri-halobutoxycarbonyl; mono-, di-, tri-halopentoxycarbonyl; and their lomed forms. The term "halo" denotes a fluoro radical,

chloro, bromo or iodo.

  The term "aralkoxycarbonyl" refers to groups

  benzyloxycarbonyl, phenethoxycarbonyl, phenpropoxycarbonyl

  nyl, phenbutoxycarbonyl, phenpentoxycarbonyl, diphenyl-

  methoxycarbonyl, diphenyloctoxycarbonyl and their forms

  isomers and the fluorenylmethoxycarbonyl group.

  The term "alkoxycarbonyl" refers to the isopropyloxycarbonyl, tert-butyloxycarbonyl and t-pentyloxycarbonyl groups. It should be noted that, for the purposes of this specification, when there is more than one hydroxy or alkoxy group on

  the sugar portion, these groups may be equal or different

ent.

  The invention also relates to a method

  chemical production of compounds analogous to the spectator

  mycin. The method of the invention thus highlights the importance of the stereochemistry of the glycosidic bond, that is to say the binding in position 1 'of compounds of formula I. The term "a-anomer" designates a substituent in

  position 1 'below the nucleus plane and the term "B-

  "anomeric" refers to a substituent at the 1 'position above the nucleus plane The term "g-anomer" refers primarily to anomers which have the C-1 configuration corresponding to

spectinomycin.

  Compounds of the invention that are endowed with desirable biological activity are e-anomers of the compound I. This glycosidic configuration meets

  in spectinomycin shown on page 1. As a result,

  therefore, a selectivity is desirable in the 1 'position is desirable to obtain biologically active analogs of

spectinomycin.

  Actinamines and actinamine derivatives

  take the aminocyclitols represented by formula VI

after.

  The term "sugars" refers to substitute pyrannes

  killed, natural and synthetic sugars, chiral

and achiral.

  The method of the invention is advantageous in that it offers this selectivity in a total chemical synthesis of antibacterial compounds which are spectinomycin analogues. So far, the prior art has not

  realized the possibility of obtaining a wide variety of

  spectinomycin in a process that produces a hemi-

  hexagonal ketal of structure similar to that of spectinomycin, endowed with biological activity, by transformation of

  delta-hydroxy-oximes to delta-hydroxy-ketones.

  Another advantage of the process of the invention resides in the fact that an initial reaction actinamine carrying protected amino groups is shown to be capable of reacting at the hydroxyl group at the C-5 position without additional protection of other hydroxyl groups y or a at positions C-2, C-4 or C-6 to produce the spectinomycin-like configuration. Thus, it is surprising to note that a difficult and troublesome protection of hydroxyl groups

  not in position C-5 on the reaction-actinamine

  It is unnecessary in the process described herein. Another advantage of the process of the invention lies in the selective production of a C 3 'carbonyl group,

  as such or in a masked or latent form, by a reaction

  elimination. The C-3 'carbonyl group is a

  very important, but difficult to obtain

spectinomycin analogues.

  The new process of the invention can be illustrated by the following reaction scheme: R '? "B R? R I VI N t Ris RR OH O II R6 R1, N 'R

R11 R12

  Wherein R is a group of the formula: R 1 R, R 1 where the variables R 1 to R 4 may be the same or different and are selected from hydrogen or an alkyl group. lower, lower alkenyl, lower alkynyl, halogeno lower alkyl, amino-lower alkyl, -OX and - (CH2) n-OX, provided that R1 and R2 are not hydroxy groups and that one of the variables R3 and R4 is a hydrogen atom, X is a hydrogen atom or a lower alkyl, lower alkenyl or lower alkynyl group, n is an integer having a value of 1 to 4; R5 is a hydrogen atom or a lower alkyl group; R6 to R14 are selected from hydrogen and lower alkyl, lower alkenyl and lower alkynyl; R5 is a hydrogen atom or a lower alkyl radical; R'7, R'8, R'11

  and R'12 are selected from lower alkyl, alkenyl

  lower nyl, lower alkynyl and a protecting group selected from aralkoxycarbonyl, halogenated alkoxycarbonyl and alkoxycarbonyl groups; provided that one of the variables R7 and R8 is always a hydrogen atom and that one of the variables R11 and R12 is always a hydrogen atom, and further provided that one of the variables R'7 and R'8 is always a protective group and that one

  variables R'11 and R'12 is always a protective group

  tor; R16 is an alkyl or aralkyl or aroyl group; A is an oxygen or sulfur atom, and B and B1 are the same or different and are selected from hydrogen and hydroxy, alkoxy, lower o-alkenyl, thio, thio-lower alkyl and lower thio-alkenyl. Some of the intermediate compounds and processes between compounds VI and II are described in United States Patent Application

N 020.073 of March 13, 1979.

  An example illustrating more precisely the process is given by the following scheme: ## EQU1 ## CBz the CBz

B OH H

CH * N the H'Hj.} {

H '

NCH3 OH

  Although the two anomers a and f can be formed in step 1 of the above process, the anomers 8 can be obtained preferentially by separation of the anomers from the a following any step of the process. Similarly, the use of an enantiomeric sugar available in the initial coupling reaction to form a high proportion of the anomer 3, or the epimerization of any intermediate or final product, may increase the amount obtained

  In addition, asteroid mixtures can themselves

  even be used as antibacterial agents in the

  measure that they contain biological activity as a consequence

  the presence of an active anomer.

  The term "anomers and asteroid mixtures of a compound" covers the spectinomycin analogs of the invention endowed with antibacterial activity. Although the $ configuration constitutes the active anomer of the invention, the expression "anomers and asterotic mixtures" has no limiting meaning, since new α-anomers can also be present without the activity of a mixture. asterisk is altered. Similarly, anomers have the analog of

  spectinomycin may in some cases be favorably

  anomerically into the active form of the analogue. Therefore, configuration a is not excluded at a stage

  any of the process of the invention.

  Furthermore, the compounds used according to the invention are products of formula I which have the S configuration because these anomers are endowed with antibacterial properties. The separation of the anomers can be carried out following any step of the process. Preferred anomers according to the invention are

  compounds bearing hydroxyl groups at the C-2 and C-positions

  6 and having the following formula:

R7 OH H

R

N OH O

  Ri4% R, 2 in which all the substituents have the definitions already given. Another aspect of the invention lies in

  new intermediate compounds used in the process.

  These intermediate compounds include: (1) anomers and asteroid mixtures of compounds of formula:

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R'7 R

N R3 R'a R9

X A

RI, 1 R'12

  (In which R is a group of formula: R 1 R 2 R 2 where the variables R 1 to R 4 may be the same or different

  and are selected from hydrogen and lower alkyl

  lower haloalkyl, amino-lower alkyl, lower alkenyl, lower alkynyl, -OX and - (CH2) n-OX, provided that R1 and R2 are not OH groups and that one of the variables R3 and R4 is obligatorily a hydrogen atom, X is a hydrogen atom or a lower alkyl, lower alkenyl or lower alkynyl group, n is an integer having a value of 1 to 4, R5 is a hydrogen atom or an alkyl group inferior; R6, R9, R10, R13 and R14 are selected from

  hydrogen and a lower alkyl radical, lower alkenyl

  lower alkynyl; R5 is a hydrogen atom or a lower alkyl radical; R'7, R R'11 and R'12 are

  selected from lower alkyl, lower alkenyl

  lower alkynyl and a chosen protective group

  aralkoxycarbonyl, halogenated alkoxycarbonyl

  genes and alkoxycarbonyl; provided that one of the variables R'7 and R 'is always a protecting group and that one of the variables R'1l and R'12 is always a protecting group; R15 is a hydrogen atom or an acyl group; R16 is lower alkyl, aralkyl or aroyl; A is an oxygen or sulfur atom, and B and B1 are selected from hydrogen and hydroxy, alkoxy, lower o-alkenyl, thio, thio-lower alkyl and lower thio-alkenyl; provided that when the compound is of the formula: wherein CBz is a carbobenzyloxy group, B1 does not

  may not be a hydrogen atom or a hydroxy group.

  (2) anomers and asteroid mixtures of compounds which have the formula: R's'

R '. B R6 R5 R1

R23 Bi / A 0R1 B'l 1 'A

- 'R1 RR

  IIIb (wherein A, B, B1, R1 to R3, R6, R '7, R8 R, R19 R' 11, R'12 R13 and R14 have the definitions given above and R16

  is a lower alkyl, aralkyl or aroyl radical, with

  that R1, R2 and R3 are not hydroxy groups).

  In compounds of formula IIIb, hemiketal

  may be wholly or partly in keto form

  open. These two forms can exist in equilibrium as follows:

MI "'% 1to

an iN 1 ORl oOH OR16 *, %% 0R1316

R R'10 R'12

R'1R R'12

11 12

  IIIb 'IIIb "New analogues of spectinomycin and

  new intermediary compounds necessary for their pre-

  paration can be obtained according to the method indicated above. This process is also a general process for preparing spectinomycin and a wide variety of

  of spectinomycin analogues. Spectinomycin is an anti-

  biotic aminocyclitol type with remarkable structure in that there is a single condensed sugar to actinamine, both by an eglycosidic bond and a hemiketal bond. The

  process according to the invention for the preparation of

  This remarkable fusion is a synthesis that combines a sugar derivative and a protected actinamine. Sugar can be a natural derivative or can be synthetic,

chiral or achiral.

  The process has two basic steps, as shown in the diagram above. Step 1 comprises the following secondary steps: CBz OH MeN OH

HO - OH

  NMe i. CBz VT2 + 3 zrQ # \ <OAc 0 /// OAc DAc I VIla DMF VaIII Key CHO CBz Hel diluted tE H w MeN1 OAc HO ', "OtAc - HMe OH OAc CBz IVa 1c, a [m CBz

CHO X

CH3OAc CBz Illa Sid CBZ i O H

CHN-X

VCH3 O

  Other examples shown below are: ## STR2 ##

ON = II

OCH3 VIlb jla CBz OH

I, H

CH3N -OAc 0 OAc HO 4: Ix OH OAc

NCH3 H OCH3

  1 Vb CBz Ib HC1 _.i, C% .C.L IIa illustrating step 1 are VIIc VIa CB z OH

I 1 H

  HO OOAc NCH 3 OH OAc I CBz Vc HCl I lb

CH 3 CHO

CBz OH

I 1

  ## STR2 ## ## STR2 ##

HCl / THF

H20, IIIb '1d OH CBz 1H

CH3N $ 7 H

HO -0O

NBCzH3 IIb CB pH H pH H

CH3N 'X-OH

NCH3 H CBz OH I i H CH3N 4o

HO O QA

  NCH3 OAc CBz IVc KHCO3 lcf / CBz OH H

CH3N O0

  HO yCH3 OAc CBz IIIc K2HPO4 ld CH30H N / CBz OH

I H

CH3N HO -CI lic

NCH 3

CBz | IIC

H OH H

CH3 & O.]

HO "HO

NCH3 H

have

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  Before stage la, the two amino groups of the actinamine derivative VIa are protected by blocking each at

  using a protecting group such as an aralkoxygen group

  carbonyl, haloalkyloxycarbonyl, aryloxycarbonyl or alkoxycarbonyl, these groups being well known for this purpose in practice. For example, you will find general information about the preparation and elimination of the group

  protector of carbobenzyloxy and carbottiobutyl derivatives

  amino acid oxy in the article by R. A. Boissonas, chapter entitled "Selectively Removable Protective Amino Group used in the Synthesis of Peptides", In: "Advances in

  Organic Chemistry, 3: 159: 190 (1963).

  The addition of tert-butyloxycarbonyl to block an amine is also given in ALDRICH technical bulletin BOC-OH (September 1976). of the

  information on the use of the trichloroethoxy group

  carbonyl to protect the amines are given by Windholz

  and co-workers in "Tetrahedron Letters", 2555 (1967).

  Actinamines can be prepared by methods well known in the art (see, for example, Suami et al.

  coworkers, "Bull Chem., Soc. 43, 1843 (1970).

  The sugars are commercially available or can be prepared by methods known in the art; a method of this kind is described, for example, by Mochalin et al., in "Chem Het Comp." 699 (1977) (English translation of KHIM

Geterotsiklsoedin, 867 (1977)).

  Step la involves a coupling reaction between actinamine VI and sugar VIIa, VIIb or VIIc. This step takes place in a solution in N, N-dimethylformamide

* or in a similar solvent, for example diethyl ether

  that, tetrahydrofuran or dimethoxymethane, sometimes in

  presence of a base. The reaction is conducted very efficiently

  in a nitrogen atmosphere at ambient temperatures and pressures as described for a similar reaction by Lemieux et al. in "Can. J. Chem." 51, 53

  (1973). The temperature range of the reaction will generally

  0 to 45 C with molar ratios of activated sugar at a concentration of 0.01 M to 0.5 M in an actinamine solvent in a solution at a concentration of 0.01 to

  0.5 M in the reaction mixture from 0.2 to 4. Preferably

  the reaction conditions involve a temperature

  at 20 ° C to 30 ° C when dimethylformamide is used as a solvent, the ratio of sugar to actinamine being between 3: 2 and 2: 3. The reaction time can range from 4 hours to a week, but is preferably included

between 24 and 48 hours.

  The oxime Va, Vb or Vc produced is generally isolated from the reaction mixture by concentration or by

  concentration and vigorous agitation with excess water.

  The resulting solid is taken up in chloroform,

  then the solution is evaporated to dryness, giving the oxime

  gross mediator. The anomers a and S can be further separated into fractions by chromatography on a column of silica gel eluted with methanol in chloroform in a ratio of 1:99 to 2:98. However, the use of means

  separation techniques, eg extraction, crystallisation

  and / or chromatography, are within the scope of

method of the invention.

  Step lb involves removing the oxime group from compound V to form a cyclic hemiketal compound IV above. The reaction of step 1b is carried out according to deoximation procedures similar to those described by Lemieux et al., In Can J. Chem., 51, 19 (1973) and by Mallams et al in J. Chem. Soc. " Perkins I 1097 (1976). For example, the deoximation reaction is conducted by dissolving either the crude oxime or oximes a and e separated from the compound V in a solvent with subsequent addition of acetaldehyde and hydrochloric acid. The initial concentration of the oxime in the solvent ranges from 0.01 M to 1.5 M, but it is preferably between 0.1 and 0.3 M and the molar ratio of

  the aldehyde to the oxime ranges from 1: 1 to 80: 1. Chlorhydic acid

  1N is added in a ratio with the oxime of about 2: 3. The reaction mixture is stirred at room temperature for a period of 3.75 hours to 5 days and, at

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  At this stage, sodium bicarbonate can be added, the additive

  followed by a stirring period of 15 minutes. Alternatively, the concentrate can be chromatographed directly so that the silica gel removes hydrochloric acid during purification. Intermediate compound IV is recovered by standard procedures as described above. A suggested method of separation is to prepare a filtrate which is evaporated to dryness under vacuum to obtain the crude product. Again, this product can be subjected to repeated fractionations on a silica gel chromatograph using as eluent a mixture of chloroform and methanol in a

  ratio of about 95: 5. By collecting similar fractions

  Then, by thin layer chromatographic analysis and then evaporating to dryness in vacuo, the individual α and S anomers of the hemiketal, covered by

structural formula IV.

  Bases other than sodium bicarbonate may be used in the deoximation of step 1b;

  however, the absence of hydrolytic conditions is

  at this stage to prevent there being a deviation from the cyclized intermediate compounds which are stable and useful in the preparation of analogs within the scope of the invention. Therefore, the bases used should be bases that do not attack acetates, i.e., hydroxides can not be used in excess. of the

  useful examples include benzoates or monohydro-

potassium genophosphate.

  Solvents that can be used include

  acetonitrile, tetrahydrofuran, ether or di-

  methylformamide. The solvent of choice is acetonitrile.

  Step lc removes one or two substituents from the sugar portion of compound IV and generates a C-3 'carbonyl group by removal of the C-4' position to give the compound of formula III. A second elimination may relate to the C-6 'atom giving the olefin C-4', C-S '. This

  operation is conducted in the presence of a base at temperatures

  from about 0 to 80C for a period of about

21 2460953

  2 hours to a week. Bases that can be used include potassium bicarbonate, triethylamine, pyridine and an alcoholate. A preferred basic system is the potassium bicarbonate / acetonitrile system. 1 to 20 molar equivalents of base can be used, but

use 1 to 10.

  The mode of conduct of stage lc also depends on

  particular protective groups carried by the sugar portion as well as by the actinamine portion of the

  intermediate compound IV. In general, the protection groups

  the sugar portion is less difficult to remove than the groups carried by the portion

  actinamine. Step lc is a new process allowing

  both to generate under mild and selective conditions the carbonyl group C-3 'important as such or in a form

latent, by elimination.

  The C-4 'and C-6' derivatives which can be eliminated are described in the patent application

  United States of America No. 020.073 of March 13, 1979.

  For example, acetates remove acetic acid, benzoate removes benzoic acid, benzyl ethers remove benzyl alcohol, halides remove a hydrogen halide. These are non-limiting examples

  elimination occurring in step lc.

  The intermediate compounds of the invention, especially the products of step lc, are compounds useful for the synthesis of various analogues. This synthesis is accomplished by changing functional groups of the intermediate compounds by known methods such as halogenation, reduction, oxidation, chain extension, and the like. In some cases of the invention, step 1c of elimination is accompanied by the migration of the substituent C-3 'on the oxygen atom to the oxygen atom of C-2'

  with production of a C-3 'carbonyl group. This behavior

  This is illustrated by the conversion of compound IVa to compound IIIa and compound IVc to compound IIIc. In other cases, the substituent at the C-3 'position on the oxygen atom does not migrate, so that a masked or latent C-3' carbonyl group is formed; they are enolic derivatives. As an example, the transformation of compound IVb * into compound IIIb is mentioned. The enol derivatives can exist in the form of hemiketals or open ketonic isomers or as

mixtures of these forms.

  The above two versions constitute new useful selective processes finally yielding spectinomycin analogs bearing C-3 'carbonyl groups. Intermediate compounds carrying a carbonyl group

  masked or latent at position C-3 'have chemical properties

  These are remarkable features which make them interesting to use for modification by known operations, for example halogenation, alkylation, acylation, oxidation, etc. Finally, the masked or latent carbonyl group at C-3 'is much more stable especially towards the bases, so that

  constitutes a part of the intermediate compounds

tiles and insulated more easily.

  The compound of formula III is separated from the mixture

  reaction by conventional operations, for example

  piting, crystallization or concentration followed by chromatography. Step id involves removal of the protecting groups at one or more positions of the sugar nucleus. Usually, these are the C-2 ', C-3' or C-6 'positions and, depending on the nature of the protecting groups, an acid and / or a base can be used. When a base is used as in the case of the conversion of compound IIIa to compound IIa or compound IIIc to compound IIc, the hydrolysis is conducted between -10 and 500 for a period of minutes to 40 hours. The conditions that are preferred include a temperature of 20 to 300C for a duration of 1

at 20 hours.

  The alcohols that can be used include methanol, ethanol and isopropanol, with methanol being preferable. Any base that does not alter the product can be used. These are sodium bicarbonate, potassium bicarbonate, pyridine, monohydrogenphosphate,

23 2460953

  potassium, triethylamine, sodium double tartrate

  and potassium, but the catalyst of choice is mono-

  potassium hydrogen phosphate. In the first step of a two-step process for the conversion of compound I1 'to compound IIb, the C6' acetyl group is selectively removed by the above basic alcoholysis conditions,

  while the methoxy group at the C-3 'position remains intact.

  Acidic catalysis can also be used to remove the protecting groups from the sugar nucleus. For example, after the C-6 'acetyl group has been removed from the

  compound IIIb 'by the use of a base as described above.

  above, the remaining C-3 'protection can be removed by subsequent treatment with an acid to form IIb. Alternatively, compound IIIb 'can be converted to compound IIb in a single step using a

acid catalysis.

  Elimination of the protecting group (s) under

  the action of an acid is usually conducted at a temperature of

  from 0 to 800 and preferably from 20 to 300 for a period of 1 hour to 3 days, preferably 2 hours to 2 days. One can choose acids used in the

  such as hydrochloric acid, para-toluene-

  sulfuric and phosphoric; the acid is preferably used

  hydrochloric. Solvents can be tetrahydro-

  aqueous furan, aqueous dimethoxyethane, methanol or

  ethanol. Methanol or tetra-

aqueous hydrofuran.

  In some cases, it is advantageous to combine step ld with step 2 by using a reaction medium for step 2 which satisfies the conditions of elimination of the

  or protecting groups specified for step id above

  above. For example, when step 2 is conducted in

  isopropanol with addition of pyridine, the intermediate compound

  diary lIla is converted into the antibiotic called spectino-

  mycin. This shows that in addition to the elimination of

  the nitrogen (and the C-4 ', C-5' saturation) which takes place under the influence of the presence of a palladium catalyst and hydrogen, the C 2 'acetyl group is

  eliminated by alcoholysis, because of the basic system of solvent.

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  In some particular cases, it is advantageous

  omit step id or part of step id so that the elimination of protection takes place in the

  biological system to release the active analogue.

  The particular conditions of step 2 of eliminating the protection of the actinamine portion depend on the particular groups, that is to say the group R'7 or R'8 and the group R'11 or R'12 which protect the amine from the actinamine nucleus. Likewise, by the suitable choice of R'7, R'8, R'11 and R'12 and by the suitable choice of the elimination conditions of the protective group or groups known in the art, a C 4 'olefin , C-5 'may remain intact or

  can be reduced during the elimination of protection.

  When this group is a benzyloxycarbonyl or aralkoxycarbonyl group, the elimination of the protection can be carried out under a hydrogen pressure ranging from -0.7 bar to +14 bar by passage over a conventional catalyst such as palladium black, palladium fixed on carbon, palladium fixed on barium sulfate or palladium fixed on barium carbonate, suspended in a solvent, for example isopropanol, absolute ethanol, acetate

  of ethyl, toluene or tetrahydrofuran.

  Alternatively, the elimination of the protection

  compounds in which R'7 or R'8 and R'11 or R'12 are alkoxycarbonyl or aryloxycarbonyl may be conducted in the presence of an acid in a solvent such as

  nitromethane and methylene chloride.

  When R'7 or R'8 and R'11 or R 12 are

  halogenoalkoxycarbonyl groups, the elimination of

  It is preferably carried out in the presence of zinc.

  Any suitable operation may be used to isolate an analog or an asteroidal mixture of a compound of formula I and the operations described herein are not limiting in any way. If the isolation is conducted under anhydrous conditions, compounds are obtained

  carrying a carbonyl group III '(formula I).

  If it is conducted under aqueous conditions,

  hydrated compounds in the 3 'position (formula I'). An opera-

  Such a reaction involves evaporation of the excess solvent and formation of a crystalline salt of the compound. These salts can be formed by using a solution of an acid such as toluenesulphonic acid, hydrochloric acid, hydrobromic acid, hydroiodic acid or other acids in a solvent such as water or methanol. , ethanol, isopropanol, ether, 1,2-dimethoxyethane and p-dioxane. Salt is

  isolated by filtration and direct crystallization or by evapo-

  solvent, followed by recrystallization in a

suitable solvent.

  Alternatively, the crude analogs can be purified by adsorption on a column of a weakly acid ion exchange resin such as "Amberlite IRC-50" or "CG-501" resin, followed by elution with a solvent such as water, methanol, ethanol, ether, tetrahydrofuran, 1,2-dimethoxyethane or p-dioxane containing hydrochloric acid, hydrobromic acid,

  hydroiodic acid and sulfuric acid.

  Similarly, the hydroxylated analog salts can be converted back into the free analogue by passing a solution of the salt in a solvent such as water, methanol, ethanol, tetrahydrofuran or 1,2-dimethoxyethane.

  on a basic ion exchange resin such as "Dowex 1-

  X8 "(OH- form) and evaporation of the eluate containing the

  analogs bearing substituents free base.

  Each step of the above process can be carried out on asteroidal mixtures of various anomers or on the desired B-anomer itself, obtained by resolution or separation at any stage of the process. The remaining steps can be conducted on intermediates B, this

  which gives the desired biologically active anomers.

  The method of choice consists in separating anomers f from the mixture resulting from step 1 of combining a sugar with actinamine and carrying out step 2 of the process on the B-anomers to produce only analogs.

  of spectinomycin that are biologically active.

  The separation of the anomers of aspartate mixtures

  This can be accomplished with obvious modifications for those skilled in the art using conventional splitting operations. For example, compound IV can be separated

  to obtain a desired component e, by chromato-

  on a column of silica gel eluted with a mixture of methanol in chloroform, in a ratio of 1:99 to 2:98. Similarly, the separation of the anomers e can be

  performed on an asteric mixture of compound V by

  fractions obtained on a silica gel chromatograph with a mixture of chloroform and methanol as eluent. Subsequent evaporation to dryness under vacuum gives a

  separated hemiketal having structure 6.

  Another particularly effective method in the context of the invention, and which is therefore preferred, is to obtain a relatively high concentration of an anomer e in step 1, ie the coupling reaction. , using an enantiomeric sugar that promotes the formation of the structure. For example, D-arabinose gives oximes and α in a ratio of about 4: 1. In addition, to avoid long and costly separation operations involving intermediate compounds or

  resulting products to obtain configuration B, the enan-

  The monomer may also be available in quantities that are inexpensive. Thus, the use of this enantiomer makes it possible to collect a substantially enriched mixture of the desired anomer which can be purified by an operation of the type described

  above or that can be used without further purification.

  Acid salts can be formed by neutralizing

  the compounds of formula I with the appropriate acid

  chosen up to a pH of less than about 7.0 and

  advantageously at a pH of about 2 to 6.

  used for this purpose include chlorinated

  water, sulfuric, phosphoric, sulphamic, brominated

  drique, etc. The acid and base salts of the compounds can be used for the same biological purposes as the

compound itself.

  The compounds of formula I inhibit the growth of microorganisms in various media. For example, the compounds of formula I which have the S configuration are active

27 2460953

  against Escherichia coli and can be used to reduce, stop and suppress the production of mucilage in paper mill circuits as a result of their antibacterial activity against this microorganism. These 0 anomers can also be used to prolong the life of Trichomonas fetus, Trichomonas hominis and Trichomonas vaginalis cultures by protecting them from Escherichia coli contamination. In addition, the anomers are active against Bacillus subtilis, so that they can be used to minimize or prevent the odor of fish or fish boxes due to this organism. Similarly, the anomers can be used to clean the benches and the equipment

  in a mycology laboratory. O-anomers are also

  effective against Klebsiella pneumoniae.

  The compounds of formula I are also effective for the treatment of bacterial infections such as gonorrhea and tumors in mammals, including

Human being.

  The compounds of the present invention are presented for administration to humans and animals in unit dosage forms such as tablets, capsules, pills, powders, granules, sterile parenteral solutions or suspensions, eye drops,

  solutions or oral suspensions and water-type emulsions

  in-oil containing suitable amounts of the compound of formula I. For oral administration, solid or liquid unit dosage forms can be prepared. For

  the preparation of solid compositions such as

  The compound of formula I is mixed with conventional ingredients such as talc, magnesium stearate, dicalcium phosphate, magnesium aluminum silicate, calcium sulphate, starch, lactose,

  gum arabic, methylcellulose and similar substances

  from the functional point of view, used as diluents or pharmaceutical carriers. Capsules are prepared by mixing the compound with an inert pharmaceutical diluent and loading the mixture into a hard gelatin capsule

28 2460953

  of suitable diameter. Soft gelatin capsules are prepared by machine encapsulation of a suspension of the compound in an acceptable vegetable oil, petroleum jelly

  light liquid or other inert oil.

  Liquid unit dosage forms for oral administration, such as

  syrups, elixirs and suspensions. Hydro forms

  Solubles can be dissolved in an aqueous vehicle also containing a sugar, flavors and preservatives to form a syrup. We prepare an elixir using a

  hydro-alcoholic vehicle (ethanol) containing sweetener

  suitable corollants such as sugar and saccharin in combination

with aroma.

  Suspensions can be prepared with an aqueous vehicle supplemented with a suspending agent such as gum arabic, gum tragacanth, methylcellulose, etc. For parenteral administration, liquid unit dosage forms are prepared using the compound and a sterile vehicle, water being preferable. The compound, depending on the vehicle and the concentration used, may be in suspension or solution in the vehicle. In the preparation of solutions, the compound may be dissolved in water for injection and the solution may be sterilized by filtration before being loaded into a suitable vial or ampoule which is then closed. Advantageously, adjuvants such as local anesthetic, preservative and buffering agents can be dissolved in the vehicle. To improve the stability, the omDosition can be lyophilized after being loaded into the ampoule and the water removed under vacuum. The lyophilized powder is then sealed in the ampoule and an accompanying ampoule containing

  water for injectables is provided for the purpose of

  the liquid before use. Suspensions can be

  parenterally following substantially the same procedure, with the difference that the compound is in suspension

  in the vehicle instead of being dissolved and that a sterili-

  can not be done by filtration. The compound D can be sterilized by exposure to ethylene oxide before

2460 53

  suspension in the sterile vehicle. A surfactant

  or wetting agent is advantageously included in the composition

  to facilitate uniform distribution of the compound.

  In addition, a rectal suppository can be used to deliver the active compound. This dosage form is of particular interest when the mammal can not be conveniently treated with other dosage forms, for example orally or by insufflation.

  as in the case of young children or people with

  capées. The active compound may be incorporated into any of the known suppository bases by methods known in the art. Examples of these bases include cocoa butter, polyethylene glycols ("Carbowaxes"), polyethylenesorbitan monostearate and mixtures of these bases with other compatible materials.

  to change the melting point or dissolving rate

  tion. These suppositories for rectal use can weigh about

1 to 2.5 g.

  The expression "unit dosage form"

  used in this memo refers to physical units

  These units are individually unit dose units for humans and animals, each unit containing a predetermined amount of active substance calculated to produce the desired therapeutic effect in association with the necessary pharmaceutical diluent, carrier or vehicle. The specifications for the new unit dosage forms of the invention are dictated by factors

  they depend directly, namely (a) the characteristics of

  outstanding properties of the active substance and the effect

  particular that is being sought and (b) the inherent limitations

  This invention is directed to the art of formulating such an active substance for use in humans and animals, as described in detail herein and in accordance with the present invention. Examples of suitable unit dosage forms according to the invention include tablets, capsules, pills, suppositories, powder sachets, lozenges, granules, cachets, teaspoonfuls, tablespoons, drops, ampoules, vials, aerosols, landfill

  grouped multiples of any of the forms

  above, as well as other forms described herein. An effective amount of the compound is used in the treatment. The dose of the compound for treatment depends on many factors as is well known to those skilled in the art. These factors are, for example, the route of administration and the potency of the particular compound. A dosage for humans of from about 2 to about 4000 mg of the compound in a single dose, administered parenterally or in the compositions of the invention, is

  effective for the treatment of tumors and bacterial infections

  riennes. More particularly, the single dose is from 5 to about 200 mg of compound. The oral and rectal dose goes

  from about 5 to about 5000 mg in a single dose. More particularly

  In particular, the dose is from about 10 to about 2500 mg of compound. The "Preparations" described hereinafter of spectinomycin analogs and intermediate compounds for obtaining them illustrate the present invention in a non-limiting manner, those skilled in the art will spontaneously develop variants of the procedures both in the analogues and in their precursors in the context of the novel compounds described as well as in the reaction conditions and the

  techniques for carrying out the process of the invention.

  For example, for each of the Preparations and

  for each of the Examples described below, stereo

  isomers corresponding to each named compound are considered

  in the context of the present invention.

  Preparation 1-O- (3 ', 4'-di-O-acetyl-2'-deoxy-2' oximino-D-arabinopyranosyl) -N, N'-dicarbobenzyloxyactinamine CBz ## STR2 ## ## STR2 ## ## STR2 ##

I I I

  CBz CBz A solution of 11.86 g (25.0 mmol) is added

  of N, N'-bis-carbobenzyloxyactinamine in 50 ml of dimethyl

  formamide to a solution of 7.20 g (27.10 mmol) of 3,4-di-O-acetyl-2-deoxy-2-nitroso-e-D-arabinopyranosyl chloride

  in 75 ml of dimethylformamide. The reaction mixture is stirred

  for 19.5 hours at room temperature under a nitrogen atmosphere. The mixture is poured into one liter of water while stirring vigorously. The aqueous phase is decanted and the solid material is taken up in 300 ml of chloroform, the small quantity of water is separated and the solvent is removed.

  under vacuum to obtain 12.6 g of a brittle white foam.

  The total aqueous phase is extracted three times with 100 ml portions of chloroform. The combined organic phases are washed with 50 ml of water and 50 ml of brine

  and dehydrated on sodium sulfate. The volume of soil

  The solution is reduced to 40 ml and this solution is added to 300 ml of water with stirring. The chloroformic phase of small volume is separated and the solvent is removed under vacuum to give 6.18 g of white foam which is united with the other portion. The analysis of the crude product by proton magnetic resonance shows that most of the DMF was removed by this method. 1.25 g (1.89 mmol, 7.6%) of the pure α-anomer of the

  5-0- (3 ', 41'-di-O-acetyl-2'-deoxy-2'-oximino-D-arabinopyrannosyl) -

  N, N'-dicarbobenzyloxyactinamine, then 2.86 g of B-anomire

  pure 5-O- (3 ', 4'-di-O-actyl-2'-deoxy-2'-oximino-D-arabino)

  pyrannosyl) -N, N'-dicarbobenzyloxyactinamine and another

32 2460953

  portion of 3.17 g of the anomer $ of purity equal to about%. α-anomer: Infrared spectrum (CHCl 3) 3550, 3400, 3070, 2980, 1745, 1686, 1675 (ep), 1484, 1449, 1364, 1333, 1235, 1167, 1026, 669 cm -1; proton magnetic resonance (CDCl3) 6 7.40 (s, 10, aromatic), 6.07 (s, 1, H1 '),, 95 (d, J = 3 Hz, 1, H3') 5.20 ( m, 7), 3.3-4.6 (m, 10), 3.12 and 3.08 (singlet, 6, NCH3), 2.12 and 2.03 (singlet, 6,

  CH3); mass spectrum (for the tetra-trimethyl derivative

  silyl) 991 (M +), 689, 673; magnetic resonance of carbon nuclei (d6-acetone) 6 170.1, 169.8, 157.8, 156.5,

  , 1, 138.1, 129.1, 128.8, 128.4, 91.7, 86.9, 79.1, 74.4,

  69.1, 68.6, 68.3, 67.8, 67.3, 60.3, 57.7, 31.5, 31.4, 20.8,

  , 5; melting point 130-140, decomposition. Spectrum of

  high resolution mass (tetratrimethyl-

  silyl): C45H73N3O14Si4 requires 991.4169;

found: 991.4190.

  B-anomer: Infrared spectrum (CHCl 3) 3500, 3100, 2970, 1748, 1686, 1672 (ep), 1486, 1449, 1370, 1337, 1235, 1167, 1107, 1024, 700 cm 1; proton magnetic resonance (CDCl3) δ 7.40 (s, 10, aromatic), 6.40 (s, 1, H1 ') 6.05 (d, J = 3 Hz, 1, H3'), 5.0 -5.5 (m, 6), 3.4-3.8 (m, 11), 3.07 and 3.04 (singlet, 6, NCH3), 2.08, 2.05 (singlet, 6,

  CH3); mass spectrum (for the tetratrimethyl

  silyl), 991 (M) 990, 976, 975, 689, 673; Magnetic resonance of carbon nuclei (d6-acetone) 6170.7, 169.9,

  157.7, 157.1, 149.1, 137.9, 129.1, 128.7, 128.3, 92.3, 85.3,

  74.7, 70.8, 69.4, 67.3, 62.0, 60.2, 31.4, 20.8, 20.5; melting point 140155 (decomposition). High Resolution Mass Spectrum (tetra-TMS), C45H73N3O14Si4 requires 991.4169;

found: 991.4229.

  Preparation O- (3 ', 4', 6'-tri-O-acetyl-2'-oximino-2'-deoxy-α-L-glucopyranosyl) -N, N'-dicarbobenzyloxyactinamine ## STR1 ## OAc HO OH ON A xOAc NCH3 OAc CBz

OH

  A solution of 7,09 g (21 mmol) of 3,4,6-tri-Oacetyl-2-nitroso-2-deoxy-α-L-glucopyranosyl chloride CH325 in 150 ml dimethylformamide and 14.22 g mmol) of N, N-dicarbobenzyloxyactinamine is stirred for 19 hours, after which no more pyrannosyl chloride remains. The solution is concentrated to 45 and the residue is dissolved in chloroform containing 1.5% methanol. This residue is placed on a column of silica gel (3 kg) wet packed. The column is eluted with a solvent of 1.5% methanol in chloroform. After using about M NOAc liters of solent, the main product is Hlu as CH OAc NCH3 CBZ a solution of the apprc7.09 g (21 mmol) of thin layer chromatography (5% methanol in chloroform) . The fractions which contain the pure product are collected and concentrated in

  dormant 7.86 g (48%) of 5-O- (3 ', 4', 6'-tri-O-acetyl-2'-

  3.4, tri - acetyl-2-nitroso-2-deoxy-d-e-L-glucopyranosyl) -NNdicarbobenzyloy-

  in 150 m of dimethylformamide and 14.22 g (30 mmol) of N, Ndicarbobenzyloxyactinamine is stirred for 19 hours, after which no pyrannosyl chloride remains. the solution is concentrated to 450 and the residue is dissolved in chloroform containing 1.5% methanol. This residue is placed on a column of silica gel (3 kg) wet packed. the column is eluted with a solvent consisting of 1.5% of ethanol in chloroform. After using about 1 liter of solvent, the main product is eluted as can be appreciated by thin layer chromatography (5% methanol in chloroform). The fractions which contain the pure product are collected and concentrated in

  yielding 7.86 g (48%) of 5-0- (3 ', 4'., 6'-tri-O-acetyl-2'-

  oximino-2'-deoxy-at-L-glucopyranosyl) - N, N'-dicarbobenzyloxy-

actinamine.

34 2460953

  DC (CH 3 OH) [() 926 mp + 18,900 + 1300 (a) 25-53 (C, acetone 0.9) Proton magnetic resonance (CDCl3): 2.02 (9H, s), 3.03 (6) .H, s), 5.04 (4H, s), 6.20 (1H, s), 7.32 (10H, s) Magnetic resonance- carbon cores

  (CD3COCD3): 170.8, 170.0, 169.8, 157.7, 157.0, 149.5, 137.9,

  129.1, 128.7, 128.3, 92.0, 85.8, 74.4, 70.4, 69.8, 69.4,

  68.6, 67.3, 62.5, 60.6, 60.2, 31.4, 31.1, 20.6 ppm.

* Mass spectrum, m / e (tetra-TMS): 1063 (M +),

1048, 793, 792, 689, 674, 645.

  Preparation lb-O- (4 ', 6'-Di-O-acetyl-3'-O-methyl-2'-deoxy-2'-oximino-B-D-glucopyranosyl) -N, N'-dicarbobenzyloxyactinamine

OH

CH3 i O C1 + oAi H OH OAc

NCH3 / 0 OCH3

CBz /

OH / DMF

CHN O O

CAc HO 'I v OAc OH OCH3 ICH3 CBz

  97.7 g (201 mmol) of N, N'-bis-carbo-

  benzyloxyactinamine to a solution of 60.8% (196 mmol) of

  4,6-di-O-acetyl-3-O-methyl-2-nitroso-a-D-glucopuric chloride

  yrannosyl (2) in 1.1 liter of dimethylformamide. The reaction mixture is stirred for 45 hours at room temperature under a nitrogen atmosphere and then concentrated under high vacuum to give a thick syrup. We pay c

2460953

  this syrup in portions in a total volume of 3 liters of ice water, with vigorous stirring. White solid substance

  resultant is filtered, redissolved in 3 liters of chlorine

  form, freed from residual water and dehydrated on sodium sulphate. Concentration of the chloroform phase gives 143 g of a tacky white foam containing about 7% dimethylformamide according to

  nuclear magnetic resonance integration reports.

  By chromatography on 3.5 kg of silica gel, carried out using a gradient of methanol and chloroform as eluent, a coarse separation of the products is obtained. The fraction containing the anomer B is crystallized in acetone to give 4.4 g of the pure isomer. Chromatography of the mother liquors gives another 1.5 g of

the anomer 3.

  Infrared spectrum: 3500, 3310, 2920, 1750,

  1690, 1459, 1240, 1175, 1105, 1403, 735, 711 cm -1.

  Proton magnetic resonance (CDCl3): 732s,, 41s, 5.11s, 3.5-4.5m, 3.3, 33s, 3.04s, 2.02s. Magnetic resonance of carbon cores (CDCl3): 170.6, 169.6, 157.7, 156.5, 150.1, 136.5, 128.5,

  127.8, 95.7, 92.7, 77.1, 73.2, 70.1, 67.54, 64.09, 56.8,

29.7, 20.8, 20.5 ppm.

  Mass spectrum (tetra-trimethyl derivative

silyl) M + 1035.

Melting point: 214-216.

  By following a similar procedure to Preparations 1, 1a and 1b, but using appropriately substituted actinamine and sugar, the oximes of

Tables I and II.

TABLE I

B H

  ## STR2 ## where: ## STR2 ## R ,! B, OH, R, CB, CH, OR, B, R, R, R

HO- H- CH2CO0

HO- H- CH2CO {

CH, 30-

HO- HS-

CH3 S-

CzR S-

HO- "CH c'-CH: R, 3_ CHCO-

ClHCo-

Rv, R, _5

H CH.0 {-

a N

C1ICH2-

BrCH2-

c C1CH2-

N BrCH2

"C1 CY2-

"O CH: OCH2-

cDCH2o-

CH = O-

"CHM? -OCH2-

zOZH4- H

O20CH2-

H- H-

C ': C- -CH2-

OCH: O-

CHC-O-

H

CH: C-O

CH, O-O_ O

H- GCH.-

"CH: C-

I

CHUCH2-

H

OCH2 -

H CH;

At CH, -

  HO- HO- HO- HO- HO- HO- H- B HO- CH30-

C2H50-

HS- CH3S-

C2H5S-

  H-HO-HO-HO-HO-HO-HO-HO- nu- HO- nO- HO- HO- HO- HO- HO- O- CH = - # m * HH OtH) -O-? EH) - 3He

- -OH)

  O -Hi mOH in H -3OcHO d 1, v neH 'nCeg H -H H -rHOOCH93 H zH) Oc H)

-2IID30-CHI

  -OeHi - rHO: HO -eH30eHD'O -zHDO3CH3 0O {::] {H

- -OH

-OH -SrHpe -sCHD

"-SH

-OH

"-OCH3

m -H-OH -OH-OH u -OH

-H -OH

O -OH

-H -OH

NH3 Z3

N II nVsqeVi

úS609M

  ## STR5 ## -: o THE - m.

_ZH3 D

a m a a

38 2460953

Preparation 2

  N, N'-dicarbobenzyloxy-5'-demethyl-3'-O-acetyl-4 '- (R) -acétoxy-

  3 "(R) -dihydrospectinomycin CBz OH CBz OH: H at ClinCH0 M MeN t: H HOtoh OAc CH3CN and Ot Ac NMe OH 0Ac HMe OAc CBz CBz

  2.32 g (3.30 mmol) of 5-O- (3 ', 4'-diol) were

  O-acetyl-2'-deoxy-2'-oximino-D-arabinopyrannosyl) -N, N'-dicarboxylic

  benzyloxyactinamine in 30 ml of methylnitrile and 0 ml (89.4 mmol) of acetaldehyde and 2.5 ml (2.50 moles) of 1N hydrochloric acid are added. The reaction mixture is stirred for 3.75 hours at room temperature,

  sodium sulfate and stirring is continued for 15 minutes.

  The mixture is filtered and the solvent is removed in vacuo to

  obtain 2.6 g of a white solid substance. By chromato-

  on 200 g of silica gel (methanol: chloroform),

  1.45 g (2.11 mmol, 64%) of N, N'-dicarbobenzyloxy-

  1-Dimethyl-3'-O-acetyl-4 '- (R) -acetoxy-3' (R) -dihydrospectinomycin as a white solid: mp: 135.0-142.9 Infrared spectrum (CHCl 3): 3450, 3050, 1748, 1684, 1881, 1447, 1362, 1333, 1238, 1160, 1050, 1020, 680 cm -1 Proton magnetic resonance (CDCl 3) 67.4 (s, 10, aromatic) 3.4-5.4 (m), 3.05 (s, 6, NCH3), 2.10 (s, 3, COCH3), 1.95 (s, 3, COCY3);

  Magnetic resonance of carbon nuclei (d6-

  acetone) 6 170.2, 169.8, 157.5, 156.9, 138.0, 129.1, 128.3,

  94.6, 91.5, 74.7, 72.4, 67.3, 67.0, 66.3, 65.6, 61.7, 61.2,

57.5, 31.6, 20.6

  Mass spectrum (for the tris-methyl derivative

silyl) 904 (M +).

39 2460953

  Preparation 2a ECT H t OAc CBz, 7 -CB 2 Stir at room temperature

  5 hours a solution of 7.00 g (9.03 mmol) of 5-O-

  (3 ', 41,6'-tri-O-acetyl-2'-oximino-2'-deoxy-a-L-glucopyran-

  nosyl) -N, N'-dicarbobenzyloxyactinamine, 7 ml of acetonitrile, 14.2 ml of acetaldehyde and 17.0 ml of 1N HCl. 60 ml is added

  anhydrous sodium sulfate and stirred for 10 minutes.

  The solid material is filtered and washed with acetonitrile.

  The filtrate is concentrated and taken up with 15 ml of mixture.

  chloroform and ethyl acetate 1: 1 and

  Spray the solution on 150 g of silica gel using the same solvent. The eluate is analyzed by thin layer chromatography (5% methanol in chloroform) and the pure fractions are pooled and concentrated to give 96 g (87%) of the hemiketal triacetate. (a) 27-520 (C 0.7, chloroform) Proton magnetic resonance (CDCl3): 2.06 (9H, s), 2.88 (3H, s), n 3.08 (3H, s, ) 4.90 (s), 5, 13 (4H, s), 7.38 6 (10H, s) Magnetic resonance of carbon cores

  (CD3CO CD3): 170.1, 169.4, 137.5, 137.4, 128.6, 127.9, 99.2,

  94.0, 82.2, 73.9, 70.6, 68.4, 68.0, 66.8, 65.6, 62.3, 60.4,

57.3, 30.0, 20.0 ppm.

Preparation 2b

  N, N'-dicarbobenzyloxy-3'-O-methyl-4 '- (R) -6'-diacetoxy-3' - (S) -

  dihydrospectinomycin CBz OH CBz OH II: H tN 0 Ac

CH1JN 'F {AC CH'CH0 CH.N

  H0'Yb oMAc CHCt (aq) C B CCNH Bz OH! rRz CBz

2460953

  1.2 ml of acetaldehyde and 0.18 ml of 1.0N hydrochloric acid are added to a suspension of 300 mg

  (0.40 mmol) 5-O- (4 ', 6'-di-O-acetyl-3'-O-methyl-2'-deoxy)

  2'-oximino-B-D-glucopyranosyl) -N, N'-dicarbobenzyloxyactinamine in 12 ml of acetonitrile and stirred for 60 hours. The homogeneous solution is stirred with sodium sulfate for 20 minutes and filtered. Chromatography of the product over 10 g of silica gel in a mixture of chloroform with

  % and ethyl acetate, 150 mg of N, N'-di-

  carbobenzyloxy-3'-O-methyl-4 '- (R) -6'-diacetoxy-3' - (S) dihydro

  spectinomycin. Proton magnetic resonance (CDC13): 7.32 s, 5.1 s, 4.68 s, 3.5 s, 3.1, 2.1 6

  Magnetic resonance of carbon nuclei (d6-

  acetone) 171.0, 170.3, 157.8, 138.2, 129.2, 128.5, 95.8,

  93.8, 84.5, 75.2, 73.1, 69.8, 67.4, 66.4, 65.2, 63.7, 61.2,

  57.9, 57.5, 30.8, 29.8, 2.10, 20.7 ppm.

  Mass spectrum (tri-trimethylsilyl derivative)

m / e (M +) 948 (M-15) 933.

  Using a procedure similar to that of Preparation 2 or 2a, but choosing the appropriately substituted precursor oxime in Tables I and II, the protected spectinomycin analogs of

Tables III and IV below.

H

"HtOOIH30 UH.

  ## STR3 ## ## STR2 ## ## STR2 ## ## STR2 ## ## STR2 ## -O-OH -SrHe:

-SCH 3)

SH

U -OCH3

m -H n -OH-OH n -OH n -OH -OH

N -OH

-H -OH

read _.

  ## EQU1 ## where ## EQU1 ##

úS609 Z

  ## STR5 ## OH-OH -OH -OH -OH -OH -OH -OH -OH -OH -OH -OH -OH -OH -OH -H-sf.Ht -SCHeD SH -OCH3 -OH - - = Ii -nj UE u Ug # # N n # N Àl

ZHDG; -0HD

  ## STR5 ## wherein R 1, R 3, OH, OH, OH, OH

-H -Oeinc 3 HDu-H.

-s -0 H

H -03CHO

t "- '

- = H3OJCHD

0O -H -OH

- - 0-H

-OH -OH -OH -OH -OH -OH

OH OH

Ht) -OH

H3 -OH

-SH -OH

He3 -OH OcHi -OH

-H -OH

-OH -H

-OH -SH :)

-OH -ScHi

-0H -SH

-OH -OrHe: -OH -ocr

-OH OH

i. cid z83 CH / - H i AI fnvsZ 'r,

úS6O9Z

H s -rH = '-3CH m -j'H:

AT - H.

  d S78 # m # - m 2plèi953 Preparation 2c

  N, N'-dicarbobenzyloxy-6'-acetoxyspectinomycin-3'-ether

Methylenol CBz CBz OH [I H

! 1 I H CH3N,

  CH'3 A KHCO3 Xl OAc H 'NO. A solution of 101.3 mg (0.136 mmol) was stirred at room temperature (CdH 3).

  N, N'-dicarbobenzyloxy-3'-O-methyl-4 '- (R) -6-diacetoxy-3' -

  (S) -dihydrospectinomycin in 5.5 ml of acetonitrile with 350 mg of potassium bicarbonate for 6 days at room temperature. We filter the mixture on "Celite" and we

  wash the bicarbonate with an additional volume of

  nitrile. Concentration of the filtrate gives 80.5 mg

  N, N'-dicarbobenzyloxy-6'-acetoxyspectinomycin ether

2'-méthylénol.

  Proton magnetic resonance (d6-acetone) 7.4 s, 5.1 s, 4.8 s, 4.75 s, 4.6-4 m, 3.9 s, 3.5 s, 2.0 s

  Magnetic resonance of carbon nuclei (d6-

  acetone) 171.1, 158.0, 154.2, 138.2, 129.2, 128.5, 95.7,

  95.5, 89.3, 75.3, 71.7, 67.4, 66.9, 66.5, 66.2, 60.8, 60.7,

  57.8, 55.6, 30.8, 20.75 ppm Mass spectrum (tritrimethylsilyl derivative)

m / e M + = 888, M-15 = 873.

  Using a procedure similar to that of Preparation 2c, but choosing the appropriately substituted hemiketals, the protected analogs are obtained.

  of spectinomycin of the following Tables V and VI.

--oN -c R

_- 'E ...-- J

  ## STR5 ##, ## STR2 ##

H -H30OCH)

9iH3

CED ZED

NOT

t k I L.).

H d A flVggV.l

úS6099Z

  -OH-OH -OH-OH -OH-OO-OH -O-O: n -OH-or -OH-OH-H -ScH 3 -OH-OH -SCH) -SH-OH -OH - $ HC 3 -StH 3 -SH -SHe -OCH3 -H -OH -OH -OH -OH -OH -OH -OH II-III-III-OH-OcH3-OrRDH-t <H r D3

H --H:) OD'H3

H TA flV 21 rsv St, CFeD

- I

  ## STR5 ## -OH-OH-OH-OH-O -OH-OH-OH -OH-OH-OH-OH-OH -OH-OH-OH-SiHr: 3-OCH 3 -SH] OsHr) -OCH) ## STR5 ## ## STR2 ## ## STR2 ## -

46 2460953

  Preparation 3 N, N'-dicarbobenzyloxy-5'-demethylspectinomycin CBz OH CBz OH MeN v1) K2CO3 lMeN 2

CH 3CN

  ## STR2 ##

  1.32 g (1.92 mmol) of N, N'-dicarboxylic acid are dissolved in

  benzyloxy-5-demethyl-3'-O-acetyl-4'-R-acetoxy-3 '- (R) dihydro

  spectinomycin in 20 ml of acetonitrile and add 1.00 g

  (7.24 mmol) anhydrous potassium carbonate. After stirring

  For 20 hours at room temperature, the mixture is filtered and the solid is washed with another 20 ml of acetonitrile. The solvent is removed in vacuo, ml of methanol is added and the solution is treated by adding 1.40 g (8.04 mmol) of potassium hydrogen phosphate. The mixture is stirred for 22 hours at room temperature, filtered and the solid residue is washed well with methanol. Removal of the solvent in vacuo gives 1.37 g of an orange-colored, gum-like solid. Fast filtration over 40 g of silica gel (ethyl acetate) gives 950 mg of a solid

  white. A new purification on prepa-

  2000μm silica gel (ethyl acetate) gives pure N, N'dicarbobenzyloxy-5'-demethylspectinomycin: Infrared spectrum (CHCl 3) 3600, 3100, 1748, 1695, 1443, 1333, 1156, 1111, 700 cm 1 Proton Magnetic Resonance (CDCl3) δ 7.40 (s, 10, aromatic), 5.13 (s, 4), 3.1-5.0 (m), 3.05 and 2.95 (singlet) , 6, NCH3)

  Magnetic resonance of carbon nuclei (d6-

  acetone) (relative to CD3COCD3) δ 201.5, 157.1, 156.8,

  137.5, 128.5, 128.2, 127.8, 97.7, 92.4, 74.4, 74.1, 66.6,

, 8, 65.0, 60.8, 56.7, 38.2, 30.8.

  By following procedures similar to

  those of Preparation 3, but using the hemiketal precursor

  suitably substituted cursor for the N, N-bis-carbo-

  benzoxy-3'-O-acetyl-4 '- (R) -acetoxydihydrospectinomycin, the protected analogs of spectator-

mycin from Tables VII and VIII.

  ## STR2 ## ## STR2 ## H _ -Hr3 H -OcHR

"-O: H3>

  ## STR2 ## ## STR2 ## ## STR2 ## ## STR13 ##

IIA nvsIgvi.

  ## STR5 ##

N. N

* H 3 ir _ (evDo-nHDOrH) aj N a M M N

H H

gw, & U-eHODOcH3 -rmoCH :)

N #H

at

N -OH

-SZHC3

-SCHD

N -SH

OCH)

0 -H

a -OH

* -OH

N -OH

a -OH a -OH

- -OH

-H -OH,

, 'at

CID 23

  ## STR1 ## in which

-SUH = 3

  ## STR3 ##

NCH3 OCH3 NH

  CBz CBZOH Cz 0.5 m of aqueous hydrochloric acid

  4 N to a solution of 25 mg (0.04 mmol) of N, N-diethyl ether.

  carbobenzyloxy-6'-acetoxyspectinomycin-3'-methylenol in 0.65 m tetrahydrofuran. The solution is stirred for 28 hours at room temperature. The reaction mixture is diluted with 25 ml of choroform and washed with 4 ml of saturated sodium bicarbonate and 4 ml of water. The organic phase is collected, dried over sodium sulfate, filtered and concentrated to give 22 mg of N, N'-dicarbobenzyloxy-6'-hydroxyspectinomyHFine.

NCH3 H: 0

  O CH3 CBz CBz Resonant addition of 0.5 magnesium hydrochloric acid (CDCl3): 7.25 s, 5.1 s, 4.75 s, 4.5-3.5 m, 3.0, 5s

  4 N A magnetic solution of carbon nuclei NN-di-

  (CDCl 3): 157.5, 156.5, 136.3, 128.5, 128.1, 127.8, 96.8,

  91.0, 77.0, 74.3, 74.0, 72.1, 67.6, 67.5, 65.8, 65.2, 64.4,

61.7, 57.1, 39.0, 29.7 ppm.

  Preparation 3b NN-dcarbenzyloxy-6-actoxyspectinomyine-3-methylnol in 0.65 ml of tetrahydrofuran. The solution is stirred for 28 hours at room temperature. The mixture was diluted with water (1 ml) and the reaction mixture was diluted with water (4 ml) and a solution of 90 mg (0.143 mmol) of hydrochloric acid. The organic phase is dried over sodium sulfate, filtered and concentrated to give 22 mg of NN-dicarbobenzyloxy-6-hydroxy-6-spectinomycin-3'-methylenol in the organic phase, dried over sodium sulphate. 2.2 moles of tetrahydrofuran urea. The reaction mixture (CDCl 3) is stirred: 7.25 sec, 5.1 sec, 4.75 sec, 4.5-3.5 m, 3.0, 6 sec Magnetic resonance of the carbon cores

  (CDCI 3): 157.5, 156.5, 136.3, 128.5, 128.1, 127.8, 96.8,

  91.0, 77.0, 74.3, 74.0, 72.1, 67.6, 67.5, 65.8, 65.2, 64.4,

61.7, 57.1, 39.0, 29.7 ppm.

  Preparation 3b N, N-dicarbobenzyloxy-6-hydroxyspectinomycin

CBE OH

CBz qH

C113N -, -OH '-O

  OH3 NCl1: H: 3 NCH,! CBiz CBz 1.6 ml of 4N aqueous hydrochloric acid is added

  to a solution of 90 mg (0.143 mmol) of N, N'-dicarboxylic acid

  bobenzyloxy-6-hydroxy-6-spectino.mycin-3'-methylenol in 2.2 ml of tetrahydrofuran. The reaction mixture is stirred at room temperature for 24 hours and treated

  as described in Preparation 3a. N, N'-dicar-

  Bobenzyloxy-6-hydroxyspectinomycin has the same mobility, in thin layer chromatography analysis, and the same properties as the product of Preparation 3a. The products are collected and chromatographed on silica gel using as eluent a mixture of ethyl acetate and

chloroform 1: 1.

  Using procedures similar to those of Preparations 3a and 3b, but choosing the appropriately substituted e-enol ethers, instead of the P-enol ethers used in Preparations 3a and 3b, the protected analogues of spectinomycin are obtained. Tables IX and XH.tFDO1D -OH -OH -OH -OH -OH -OH -OHR

-OH OH

S -SeHZ.-OH -ScHD -OH

-SH -OH

At -SH'r -OH

-OCHO -OH

-H -OH

-OH -H

-OH -SH = 3

-OH -ScH5

-OH -SH

-OH

-OH -OCHD

-H -OH -OH

CHD Z93

  N H tod XI nVsiovi úS609tZ H Fr :) H ZI -cHDOH H Oc3

-0 CH)

  - "HDO" H) -Hf -t0O .: ## STR2 ## II. r .. HDO. ## STR2 ## ## STR4 ## ## STR2 ## ## STR5 ##

- -OH OH

## STR5 ##

.H .: -SH -OH

- --H3 -OH

"- -OCH -O H

. u -H -OH

"". -OH -H

- -OH1r -OCH

-. -OH-5OH:

. 4 -OH -SCH1

4 4 -OH -SH

4 to 4 -OH -OH3

. . -OH -OCH3

HO 'HDOH

H -tOHOH -H -OH -OH cH3 Z83

O O N

H d x esaV

ú56MZú

ES

úS609îZ

  Preparation 3c N, N'-dicarbobenzyloxy-2'-O-acetyl-4 ', 5'didehydrospectinomycin - OH CBz OH

CH H

KHCO3 CH3N CH3

  ## STR13 ## 14.58 g (146 milli-equivalents) of anhydrous potassium bicarbonate and 384 ml of acetonitrile are heated in a flask equipped with a distillation flask. When

  ml of solvent distilled, allowed to cool

  at room temperature under a nitrogen atmosphere the remaining suspension, then the hemiketal triacetate of Preparation 2a is added. The mixture is stirred at room temperature for 41 hours; after this period, the starting material is absent as can be appreciated by thin layer chromatography (5% methanol in chloroform). The solid is filtered, washed, and the filtrate is concentrated. The residue is taken up in 40 ml of chloroform and the inorganic substances are filtered. The filtrate is concentrated giving 14.51 g of a foam. After dissolving the foam in chloroform and cooling, a mass (7.55 g) is formed which is recrystallized from 20 ml of

  chloroform giving 3.42 g of N, N'-dicarbobenzyloxy-2'-O-

  acetyl-4 ', 5'-didehydrospectinomycin melting at 149-152. Chromatography of the mother liquors collected on 800 g of silica gel, with a mixture of methanol and chloroform 1: 4, gives another 3.37 g of product (total yield of

55%).

  Ultraviolet spectrum (C2H50H): 204.5 nm

  (20,950), 206, shoulder (20,650), 267 (11,550).

  DC (CH3OH) LJ3ma10 -22,000, 1) max +33,500,

() 7D-43 (C, 1, acetone).

  Proton magnetic resonance (CDCl3): 2.07 (3H, s), 2.15 (3H, s), 3.08 (6H, s), 5.12 (4H), 5.40 (1H, s) ,, 95 (1H, s), 7.32 (10H, s). Magnetic resonance of carbon cores

(CD3COCD3):

  182.5, 172.9, 169.5, 137.6, 137.5, 128.6, 127.8, 102.7, 95.5,

  93.1, 75.0, 74.0, 66.8, 65.6, 60.3, 59.6, 56.7, 31.1,

  20.4 ppm. Exact Mass: 784,3097; the formula C38H52N2012

requires a mass of 784.3059.

  Using a procedure similar to that described in Preparation 3, but replacing the hemiketal obtained in Preparation 2a with the appropriately substituted hemiketal, protected analogues are obtained.

  didehydrospectinomycin in Tables XI and XII.

  [,; 3nq -a s a [XuotidoJdos.t O * PtHD cHi C H3 of H :) O -cX3 n -; O -X 3CHO {1 OoH i. zGD DCH lN1. ## STR2 ## wherein R 1 is -OH-OH -OH-OH -OH-OH-OH-OH -OH-OH-OH -OH-OH-OH -OH-OH-OH-OH -OH ## STR2 ##

-IH.OO-H3

  -cm10 - H3 -el. -CH3 -CH3 -eH3 -CHM -cH3 -CHD H

TABLE XII

  CH3-CH3-CH3-CH3-CH3-CH3-CH, -CH3-CH, -CH-CH3-CH3-CH3-CH:

C2H, _-

  CH3 CH, C_2H? C n), CH3-II CH3C-CH1-CHC-CH3C-CH3-CHIC-CH3C-CH3 CHC2-CH3O-

CH: CO-

o

CH2CH2.

CH.C.q: C CH3 (CH2): t

CH (CH2) C

CH3 (CH2) C

  sec-butyryl isorropionyl. tarti o-butyryl B B., HOCH: O-

C2H50-

HS- CHjS-

C2H4S-

  11- HO- HO- HO- HO- TOHO- HO- HO- HO- HO- HO- HO- HO- HO- HO- HO- HO- HOHO- HO- HO- HO- HO- HO- HO- H- Ho- CHS-

C: HSS-

  ## STR4 ## Preparation 4 N, N'-biscarbobenzyloxy-4 ', 5'didehydrospectinomycin CBzOH. CBz H CH3NMeOH CHs

K2HP04 '

NCH

NCH 0 ÈN-CH3 0H

OCCH3 Cz CBz i-CBz o

  1.0 g of N, N'-biscarbobenzyloxy-2'-O-

  acetyl-4,5'-didehydrospectinomycin to a suspension of potassium monohydrogenphosphate (0.40 g) in 20 ml of anhydrous methanol and stirred at room temperature for 1.5 hours. The solvent is removed under reduced pressure and the organic material is dissolved in methanol at room temperature.

  1.5% in chloroform, then the chromato-

  on 225 g of silica gel using 1.5% methanol in chloroform. Fractions containing the product

  are pooled and concentrated in 0.51 g (55% N, N'-bis).

  carbobenzyloxy-4 ', 5'-didehydrospectinomycin.

  DC (CH3OH): (3ma4 -8300 + 2100, ([) 26a6 + 10 500 + 2100 (aoD -56 (C, 1.0, CH3OH) Magnetic resonance of carbon cores

(CD3COCD3):

  187.6, 175.8, 157.2, 138.1, 128.4, 101.7, 99.3, 87.7, 76.3,

  64.6, 63.8, 67.3, 66.7, 66.3, 65.3, 60.8, 60.0, 31.5,

21.3 ppm.

  Mass spectrum: m / e (tri-TMS); 814 (M +), 799

(M-15).

  Using a procedure similar to that

  of Preparation 4, but replacing the N, N'-biscarbo-

  benzyloxy-2'-O-acetyl-4 ', 5'-didehydrospectinomycin by the appropriately substituted didehydrospectinomycin derivative, the protected analogues of didehydrospectinomycin are obtained

tables XIII and XIV.

  Embedded image -OH -OH -OH -OH -OH -OH -OH -S, -H -S = H3 -tH -QeHe: -OCH) -OH -OH -OH -OH

CED 293

  ## STR5 ## wherein Y is CH 2 OH 3 OH-OH 3 -OH. Embedded image ## STR2 ##

CHO --OH -H

HZ -OH -OH

11c3z -OH -OH oH3, c-O -OH

-OH OH

-OH OH

c -XScH) -OH-O- '

-HO -S'HO -0

-cH 0 -CH3 -'Hz -rH -dei

-CHD -c'H "-OH

  CH 2 OH-CH 2 OH-CH 3 OH-CH 3 OH-OH-CH 3 OH -SH 2 -HH 3 OH-OH-OH -SH-OHD-OH-SHR-CID-OH -OcHO

-CH3 -OH -OH

HN2 CHH

SH0 ZE

AIX nlVS'IsE.

úS6099

  PREPARATION 5 N, N'-Dicarbobenzyloxy-6'-hydroxyspectinomycin-3'-methylenol CBz OH ## STR1 ## wherein: ## STR1 ##

I NCH3

CBz I CBz

  103.9 mg (0.154 mmol) of ether of 3'-

  N, N'-dicarbobenzyloxy-5'-acetoxyspectinomycin methylenol in 6.8 ml of methanol and the solution is shaken with 115 mg

  of potassium bicarbonate for 24 hours at room temperature

  ambient temperature. The mixture is concentrated, redissolved in chloroform, the solution is filtered and concentrated again to obtain 90 mg of N, N'-dicarbobenzyloxy-6-hydroxyspectinomycin 3'-methylenol ether. No acetate methyl groups are observed in the proton magnetic resonance spectrum or the resonance spectrum

magnetic carbon cores.

  Proton magnetic resonance (CDC13) 7.31 s,

5.10 s, 5-3.75 m, 3.5 s, 3.16s.

  Magnetic resonance of carbon cores (CDCl3): 158, 152.5, 136.5, 128.2, 95.1, 94.9, 88.7, 77.2,

67.5, 67.2, 55.0, 34.1, 29.72 ppm.

  Mass spectrum (tetra-trimethyl derivative

silyl) m / e (M +) 918, (M-15) 903.

  Using a procedure similar to that of Preparation 5, but choosing the appropriately substituted enol ethers, the protected analogs are obtained.

  of spectinomycin in Tables XV and XVI.

-e: HDOH ZS

HDR30.

-SCH 3

  -SH -5H3 -OcH3I -H-OH-OH-OH -OH-OH-OH-OH-S-Os

H3 Z83

\ N AX NVsyT,

úS609MZ

  I. It is I OR OR i. ## STR2 ## ! úS609tz is yours - S '- 4e -oH

-SuH: -OH

,, - $ SH3 -OH

-SH -OH

-s'. -sHt1 _0S -OcH) -OH

-H -OH..DTD: "," - OH4 -OH -H

",, -OH -SOHZD

",," -OH -StH3

",., -OH -SH

. -OH -05HZ3

",, *" - OH -OcH)

- = HDOH -H -OH -OH

- O

O HD zyOa END O H

fo u 0.

IAX fnV.Lss T ú9

Example 1

  Spectinomycin H CBZ OH H OH OH CH3

I 11 0 CH3N w.

CH3.N Pd / BaSO

2 hours

NCH3 -H0 NCH3 OH0

I I

  CBz H Spectinomycin 480 mg of 10% palladium fixed on barium sulfate and 0.48 ml (5.9 mmol) of pyridine are added to

  solution of 480 mg (0.80 mmol) of N, N'-biscarbobenzyloxy-

  4 ', 5'-didehydrospectinomycin in 40 ml isopropyl alcohol

  pylique. The mixture is hydrogenated at atmospheric pressure for 5 hours, filtered and the filtrate is concentrated.

  under vacuum. The residue is redissolved in isopropyl alcohol.

  The solution is treated with 2.0 ml (2.0 mmol) of a 1 N solution of hydrogen chloride in isopropyl alcohol. By removing the solvent in vacuo, 440 mg of white solid is obtained. Recrystallization from aqueous acetone gives 164 mg (0.33 mmol, 41%) of spectinomycin pentahydrate dihydrochloride having the same physical and biological properties as the product.

natural.

  By applying the same procedure to acetate

  of enone obtained in Preparation 3c, the dichlo-

  of spectinomycin pentahydrate crystalline, in a yield of

35%.

Example 2

  -methylspectinomycin CBz OH H H H HH MeN Ht t MeN OO H0 t Pd / C H0O NMe NMe OH0

I H

  CBz A solution of 333 mg (0.57 mmol) of N, N'dicarbobenzyloxy-5'-desmethylspectinomycin in 5 ml of isopropyl alcohol at 300 mg of 10% palladium fixed on

  carbon in 45 ml of isopropanol in a Parr bomb.

  The solution is treated with 1.36 ml (1.36 mmol) of 1N hydrogen chloride solution in isopropyl alcohol and the hydrogenation is carried out for 16 hours under 0.28 MPa pressure at room temperature. The material is filtered and the catalyst is washed with isopropanol. Washing the catalyst with an additional volume of water gives more product. The material is concentrated to dryness under vacuum and the water-soluble portion is separated off. By elimination of

  solvent under vacuum, a white residue is obtained which is recreated

  tallise in aqueous acetone to obtain 112 mg of 5'-

  desmethylspectinomycin in the form of a white solid: mp 196-205 (decomposition); mass spectrum m / e (penta-TMS derivative) 678.3378; the

  Formula C28H62N2O7Si5 requires 678.3403; magnetic resonance

  carbon nuclei (D30) (compared to external TMS)

  95.3, 94.9, 93.6, 71.1, 67.4, 67.0, 62.9, 62.5, 61.0, 59.9,

, 9, 32.2, 31.7.

Example 3

  6 '-hydroxyspectinomycin CBz OH H H OH H

IBZ HI 'H

  tCH3N 'OH CHN isooDroDan lV HO) CHa Black of v HO CH3 OH CBz H

  36 mg of N, N'-dicarboxilate are shaken.

  benzyloxy-6-hydroxyspectinomycin with 20 mg of palladium black in 35 ml of isopropanol under hydrogen pressure of 0.28 MPa for 2.25 hours. The catalyst is filtered and washed with isopropyl alcohol. The filtrate is concentrated to a total volume of 10 ml and 0.12 ml of acid is added thereto.

  1.0 N hydrochloric acid in isopropyl alcohol. By concentration

  solution, a white residue is obtained which is redissolved in D20 to measure the resonance spectrum of 13C nuclei. By recovering the sample used in this

  measurement, 14 mg of material are obtained after lyophilization.

  This material is crystallized in water and acetone in

  giving 6'-hydroxyspectinomycin.

  Melting point 201-205 (decomp.) Magnetic resonance of carbon cores (D20)

* 94.9, 93.2, 73.6, 70.7, 67.2, 64.6, 62.7, 60.8, 59.6, 37.0,

  31.9, 31.4 ppm Rf of spectinomycin, 0.5 Rf of hydroxyspectinomycin, 0.25

  The immersed disc test of 6'-hydroxy-

  Raw spectinomycin shows antibacterial activity. Using similar procedures

  those of Examples 1, 2 and 3, but replacing the N, N'-bis-

  carbobenzyloxy-4 ', 5'-didehydrospectinomycin, N, N'-bis-

  carbobenzyloxy-2'-O-acetyl-4 ', 5'-didehydrospectinomycin, the

  N, N'-dicarbobenzyloxy-5'-desmethylspectinomycin and N, N'-

  dicarbobenzyloxy-6'-hydroxyspectinomycin by the spectator-

  appropriately substituted protected mycine, spectinomycin analogues shown in Tables XVII are

and XVIII.

-H3 -H

-H -tH30H

-H -: H30: H3

-H3DOH -H

-tHDQH -H

HD -H

- HDOH -H

-HDOH 'H

= H30H -H

cHDOH iH

-H30H -H

-HDOH 'H

-tHDOH -H

-CHD0H -H

-CH30H -H

- = HDOH -H

-H:) OH -H

AT HOH -

-OH OH

-OH OH

-OH OH

-OH OH

-OH OH

-OH OH

-OH OH

-s "$ Hc3 -OH -ScH: 3 -OH

-H -OH

-CHei -OH

PCHD -OH

-H -OH

-OH -H

-OH -S.H: D

-OH -SCH3

-OH -SH

-OH -OH-3

-OH -OCH: 3

-OH OH

iS

H 3'H

HO s, H N ú9 IIAX nlvsqIevl L9

úS60M9

-H

-H 'HDOH

-H -e3H -cHO

-HDOH -H

  -H rH3Gn -, HeOH -H-th'05 -H -cX30 -N

-ZH3OH -H

-H chDOH -H

2: .0H -H

-.th: OH -H -tHDOH -H

-'H = OH -H

-rHAOH -H - = fOOH -H

= HOXH -H

- = H3OH -H

-cNOOH -H

CXIOH -H

--r- -b

-OH OH

-OH -H

-OH OH

-OH OH

-OH OH

-0H -Or

-OH OH

- Sr- = D -OH ch -OH

HXS -OH

-Ocho -OH

- -OH

-OH -H

-OH -SeHrO -OH -SeHr

-OH -SH

-OH -OrHC3 -0H - -OcHD

-OH OH

-i-s 8

ItIIAX lMIYS.

úú609yz-

Example 4

  4 ', 5'-Didehydrospectinomycin dihydrochloride

OH OH

OH CBz H, H OH

CH3N 2 H

o H2 OH0.2HC1 HO HCl HO

NCH3 NCH3

  CBz H is hydrogenated under pressure of 0.21 MPa during

  1 hour and a half a mixture containing 93 mg of N, N'-biscar-

  4 '-benzyloxy, 5'-didehydrospectinomycin, 70 mg palladium

  10% dium fixed on carbon, 30 ml of isopropyl alcohol

  and 0.1 N hydrochloric acid in 2 ml of isopropyl alcohol

  pylique. At this stage, another 30 mg of catalyst is added and

  1.0 ml of 0.1 N hydrochloric acid in isopropyl alcohol is

  and the hydrogenation is repeated for 2 hours and a half. The solid substance is then filtered and the filtrate

  is concentrated to give 50 mg of 4 ', 5'-dihydrochloride

  dehydrospectinomycin in the form of a solid. This product is active against E. coli and does not contain spectinomycin or dihydrospectinomycin, as shown by various layer chromatography systems

  (CHCl3: CH3OH: NH4OH-3: 4: 2) (NH4OH at 3% in

  tone). In this system, it presents an active task po) which

  corresponds to a reference standard of 4 ', 5'-dehydrospectino-

  enantiomeric mycine having the above characteristics

  after: Mass spectrum (tri-TMS): m / e 546 (M +), 531

  (H-15), 492, 426, 401, 361, 271, 199.

  Magnetic resonance of carbon cores (D20, external TMS): 189.6, 179, 3, 102.8, 98.5, 72.6, 69.9, 67.0,

  66.5, 62.6, 61.5, 59.7, 32, 32.1, 22.0 ppm.

  Following a procedure similar to that of

  Example 4, but replacing the N, N'-biscarbobenzyloxy-

  4 ', 5'-didehydrospectinomycin by 4', 5'-didehydrospecti-

  nomycin suitably substituted, the 4 ', 5'-di-

  dehydrospectinomycins of the following Tables XIX and XX.

úS609Z

-COH - -OH

CHCO

-OH -CH

-icHc

-OH OH

-OH OH

-gii'i -cHDOH

-OH OH

-rSs0e -rt O -CHD

-OH OH

-OH

-0H -OH

_-CHDs-H

-CH -OH -OH

-CH3 -OH -OH

-cH: D-cH O -'HD -10H -HUi

-CH3 -OH -H

-CHD -OH, SCH2

## STR5 ##

-CHO -OH -OH

_cHD -o-O t H

RHO O

  L; ## STR5 ## ## STR2 ##

: HDOH -H -OH

-OHIOH-OH -OH

H '= D OH-H H

  ## STR5 ## wherein OSCH-OH-OH-OH-OH

_CH3 -OCH -OH

-.Hi -H -OH

-CHO O -OH-OH

-sHz * -OHSH

-'HD -OH-SH

-'HD. -OHçt

-CH -OH -OH

_c- '-OH -OH

-OH '8H 8C

CHH "H-H

  oH3 -O 'CH3 xx ndsqúv TL úS60otZ 72.

Claims (2)

  1. - Process for the production of anomers and asteroids of a compound of formula: ## STR2 ## characterized in that it consists of: (a) converting a compound of formula: R'7 B R6   VI to a compound of formula: ## STR2 ## and (b) removing the protection of the compound formed in step (a); in these formulas, R is a group: 73. R, R, R2 R3 R4 where the variables R1 to R4 may be the same or different and are selected from hydrogen and a lower alkyl, halogen lower alkyl, amino lower alkyl radical; alkenyl   lower alkynyl, -OX and- (CH2) n-OX and their iso-   mothers, provided that R1 and R2 are not radicals   hydroxy and that one of the variables R3 and R4 represents   preferably a hydrogen atom, X is a hydrogen atom or a lower alkyl, lower alkenyl or lower alkynyl radical, n is an integer having a value of 1 to 4; R5 is a hydrogen atom or a lower alkyl radical; the variables R6 to R14 are chosen from hydrogen and a lower alkyl, lower alkenyl and lower alkynyl radical; R5 is a hydrogen atom or a lower alkyl radical; R '7,   R'8, R'11 and R'12 are selected from an alkyl radical of less than   lower alkenyl, lower alkynyl and a group   a protector selected from an aralkoxycarbonyl, alkoxy   halogenated carbonyl and alkoxycarbonyl; provided that one of the variables R7 and R8 is always a hydrogen atom and that one of the variables R11 and R12 is always a hydrogen atom; and provided further that one of the variables R'7 and R'8 is always a protecting group and that one of   variables R'11 and R'12 is always a protective group   tor; A is selected from an oxygen atom and a sulfur atom; and B and B1 are the same or different and are selected from hydrogen and hydroxy, alkoxy, lower o-alkenyl, thio, thio-lower alkyl and lower thio-alkenyl.   2. - Method according to claim 1, characterized   in that the compound prepared is 3,5'-desmethyl-   spectinomycin, 4,5'-didehydrospectinomycin or   4,5'-didehydrospectinomycin dihydrochloride.