BE604099A - - Google Patents

Description

       

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  New antibacterial agents.



   The present invention relates to new and useful tetracyclines and the process for producing them. More particularly, the present invention relates to 6-epi-6-deoxyteracyclins, their D-substituted analogs, their 11a-substituted analogs, and their D-11a-substituted ring analogs, as well as to the manufacturing process of these compounds.



   According to the present invention, it has been found that the hydrogenation of 6-deoxy-6-demethyl-6-methylenetetracyclines, comprising various analogs substituted on the D ring and in 11a-, is a very valuable process for producing 6-. known deoxytetracyclines and furthermore, produces a class of new and useful derivatives with different chemical, physical and biological properties, of these known 6-deoxytetracyclines as a result of configura-

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 modified stereochemical tion.

   These new tetracyclines have defined microbiological activity against a variety of Gram-positive and Gram-negative microorganisms and are correctly designated as 6-epi-6-deoxytetracyclines since the steric configuration of the 6-methyl group is the opposite of that. known 6-deoxytetracyclines. Therefore, the "6-epi" nomenclature employed herein fully conforms to the recognized nomenclature for known 4-epitetracyclines.



   According to the process of the present invention, a suitable tetracycline compound defined below is dissolved or suspended in amphoteric form, or in the form of a salt obtained by addition of acid or in the form of a complex salt with a polyvalent metal, in an inert reaction solvent in the presence of a catalytic amount of a catalyst which is a noble metal and brought into the presence of hydrogen at a suitable temperature and pressure, until the reduction of the 6- group methylene takes place.



  Thereafter, a mixture of 6-epi-6-deoxytetracycline and 6-deoxytetracycline can be recovered by the usual methods including separation of the catalyst and recovery of the solvent. This mixture can then be subjected to chromatography or other methods of separating such a mixture into its components.



   The term "inert reaction solvent" as employed herein relates to any medium which is a suitable solvent or suspending agent for the antibiotic reagent, which is stable under hydrogenation conditions and which does not alter the efficiency of the catalyst. and which does not react with the antibiotic. Polar organic solvents are generally suitable and include lower alcohols such as methanol, ethanol and butanol, etc .; water soluble, cyclic or open chain ethers, such as dioxane, tetrahydrofuran, diethylene glycol monomethyl ether, 2-ethoxyethanol, lower monocarboxylic acids such as acetic acid, propionic acid, media aqueous containing the above solvents, dilute aqueous hydrochloric acid, etc.

   Note that these and other solvents are

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 rants in the known hydrogenation techniques applied to antibiotics containing tetracyclines and thereby, are irrefutable. Better results are, however, obtained when acidic media are employed instead of basic media which may cause decompositions and thus reduce the yield of the desired product. The use of the best solvents is illustrated by the examples below.



   Temperature is not more critical in the present process than it is in other hydrogenations of antibiotics of the tetracycline series. In fact, the best temperature range is approximately 0 to 60 C, the best temperature in this range is 10 to 50 C and preferably at room temperature.



  For temperatures below 0 C the reaction is excessively slow and for temperatures above 60 C decompositions of the starting material may occur. As you would expect, the higher the temperature, the faster the reaction.



   The noble metal catalysts used in the present invention include platinum, palladium, rhenium, rhodium, and ruthenium, either with or without support, as well as their known catalytic compounds such as oxides, chlorides, etc.



  Examples of suitable catalytic supports are carbon, silica and barium sulfate. The catalysts can be pre-prepared or prepared in situ by prereduction of an appropriate salt of the catalytic compound. The best catalysts are, for example, palladium on carbon 5%, platinum on carbon 5%, rhodium on carbon 5%, platinum chloride, palladium chloride, platinum oxide and ruthenium oxide. . Products like these, in which the metal is in a combined, non-elemental form, generally require prereduction before hydrogenation can be done.

   This is done simply by suspending the catalytic precursor in the hydrogenation medium, hydrogenating it, adding the substrate and continuing.

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 EMI4.1
 1-lhyd.ro-zena-uion. On the other hand, all the components can be incorporated at once at 2 "xyàrog, en-tion can be co: aEr.cëc. Cc process F- 1, F - ,, - zznT, & this to allow 1" ipëraicùr to deter .aine r:; ec.i't'- ment the quantity of hydrogen absorbed during the prereduction, catalytic and during the hydrogenolysis phase. The advancement of the hy-
 EMI4.2
 Drogenation of the antibiotic can then be controlled more easily.



   The expression "catalytic amount" employed herein will be readily understood by those skilled in the hydrogenation of known tetracyclins, it is the less amount that that employed in
 EMI4.3
 exesplep yes follow.



   Rhodium is the best catalyst for the process of the present invention because it gives the highest overall yield.
 EMI4.4
 in 6-tpi - céoY; - and c-âéso: ytëtr4cyclines. In addition, 1-1e - i ,, law called rhodius allows the maintenance of a 7-halogen substituent when the starting material is', for example, 7-halo-6-deoxy-6-draetryl-6mePylene- 5-of-, ëtrüc; cline. When it is not necessary to preserve a 7-haloenea substituent, it is preferable to employ palladium as a catalyst. Nevertheless, the other noble lethal catalysts are perfectly suitable for obtaining 6-epi-E-.àëso- and 6-deoxtetracyclines.



   The pressure employed during the hydrogenation is not critical but depends mainly on the capabilities of the apparatus.



  In general, the pressures employed range from atmospheric pressure to pressures of 2,000 pounds / square inch (140 kg / cm2). As is known, hydrogenation at atmospheric pressure is generally carried out in an apparatus in which a determined volume of hydrogen contained in a reservoir is connected to a manometer so as to be able to measure the quantity of hydrogen consumed. A bottle of citrate or magnesia and a mechanical stirrer with a calibrated pressure gauge can also be used, or a high pressure autoclave with agitation.



  For the process of this invention, the starting materials

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 suitable cO.:lprt.:nnt::rd "those yes ront. r; prt, senL: 1; pe.r 1 ::! c, -.,. ':' \ .. 5 following:
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 EMI5.3
 where Y may be hydrogen or hydroxyl, X may be hyaline, chlorine, iodine or bromine, X1 may be hydrogen, arrain or lower alylamino, X2 may be hydrogen or nitrogen, and Z may be chlorine or fluorine. In addition, the salts obtained by adding acid and the complex salts of polyvalent metals of these compounds can be used. Note that the steric configurations of positions 5a and 11a are shown in the second formula above.



   Since the start products of the process of the present invention described above are relatively new well known tetracycline compounds, Examples I to XXXI are given below to illustrate their preparation and to provide the necessary starting materials. In general, the method of preparation comprises treating an 11a-halo-6,12-hemiketal of a suitable tetracycline with a strong acid of the dehydrating type such as sulfuric, trifluoroacetic, polyphosphoric, perchloric, hydrofluoric or other acid. of the same kind. Among these, hydrofluoric acid will be preferred. Optimum conditions are usually determined by routine.

   In general, the selected 11a-halogenated hemiketal is simply added to the selected acid and allowed to react at a temperature best between 0 and 50 C for a period of time.
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 stretching a hard '

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 hilarious of one here several deads. When the reaction is complete, the product is suitably isolated, that is to say, in the case of volatile acids, by evaporating these to obtain the residual product, and in other cases by usual methods such as, for example, by stirring with a solvent in which the product is not soluble (for example diethyl eter) in order to precipitate it.

   These O-methylene compounds can be converted before hydrogenation to salts, such as hydrochlorides, by the usual methods.



   Variations in substituents of the starting materials of the process of the present invention are also illustrated in the examples below.



  Briefly, the 7-halogenation of the starting material can be obtained by the direct halogenation of a 11a-halo-6-deoxy-6-demethyl-6-methylene-5-oxytetracycline by well known reagents such as N-chloro- , N-bromo- and N-iocosuccinimide. tour rette realization, it has been found that liquid hydrofluoric acid is an excellent reaction medium and that temperatures ranging from -40 C to 60 C are perfectly suitable. The resulting 7,11a-dihalogen compound can be used as such or can be converted to the corresponding 11a-dehalogenated derivative by the methods illustrated in Example VI.



   Direct nitration of the selected 6-deoxy-6-demethyl-6-methylenetetracycline is suitable for preparing the 9-nitro starting materials of the present invention. The starting material is reacted with nitric acid by itself or formed in situ, for example with potassium nitrate and sulfuric acid. The best results are obtained when the nitration is carried out in a solvent such as liquid hydrofluoric acid, lower alkyl acid, etc. When hydrofluoric acid is used as a solvent, it can also act as a proton donor and form nitric acid in situ with nitrates.

   It is necessary to avoid raising the temperature above 50 C, the best results being obtained at room temperature and it is necessary to work even at lower temperatures when using hydrofluoric acid as a solvent.

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 Reaction times can vary from 15 minutes to 12 hours.
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  When the desired starting material is 11a-cesnalo-6-deoxy-b-demethyl-b-methylenetetracycline, the 11a-dehalogenation can be carried out by either chemical or catalytic reduction using methods well known to those skilled in the art. Example XXXVII below illustrates the reduction by hydrogenation of a
 EMI7.2
 lla-chloro-6-deoxy-6-deaethyl-6-methylenetetracycline To obtain the corresponding lla-dechlorinated derivative and also shows that the lla-fluoro group is not susceptible to reduction by this process. On the other hand, the lla-fluoro groups are removed in the desired starting products by reduction methods illustrated in Example IV, method D, in which the reduction of the powdered zinc and of the dilute hydrochloric acid is employed day. .
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  The preparation of the products of âtaart 9-to: xii; or of the process of the present invention can be made by reduction of the corresponding 9-nitro compounds using the methods generally employed for the conversion of nitro compounds to amino compounds.



  Such methods are illustrated by Examples XIII, XIV and XXX. The acylated 9-amino starting compounds of the present invention are
 EMI7.4
 sussi prepared by the procedures customary in this case. The best method comprises the acylation of 9-nino-E-3esoxy-bdemethyl-6-sethylenetetracycline by the desired diacid anilydride or by an equivalent thereof such as an acid chloride in the presence of a base, for example a tertiary amine such as pyridine, in an inert solvent. Examples XVIII and XIX, among others, illustrate these acylation techniques.



   As stated above, reduction of the 6-methylene moiety of a starting compound selected in accordance with the present invention can produce the removal of a 7-halogen substituent and produce the removal of a. substituting lla-chloro. If we want
 EMI7.5
 that such substituents exist in the.: J.éla! 11> t: final or in each of its components, for example, the 6-epi-esoxyLetracy "cline

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 or 6-deoxytetracycline, they can be reintroduced by the chlorination technique described above and illustrated by Examples XLI-XLV. Further, assuming that a 11a-fluoro starting material is useful during reduction of the 6-methylene group, this substituent can then be removed by the method described in Example XXXVIII.



   The choice of a 9-nitro-6-deoxy-6-demethyl-6-methylene-tetracycline as the starting material for the reduction process of the present invention will cause the reduction of the 9-nitro group to 9-amino. Therefore, if the 9-ni ne of normal or epimeric configuration is desired, the 9-nitro moiety should be introduced into the product of the reduction process of the present invention by the same nitration means as described herein. above and illustrated by Examples L, LIV, LV, LVI and LVIII.



   In order to stabilize the 6-deoxy products of the process of the present invention in an acidic medium, the substituted 9-amino derivatives such as 6-epi -6-deoxy-9-amino-5-oxytetracycline can be converted into the 9- compounds. corresponding chloro. This process comprises diazotization followed by a conventional Sandmeyer reaction and illustrated in Example LIII. It will be appreciated later that this Sandmeyer reaction can be applied to the preparation of other halogenated derivatives, nitriles, etc.



   Direct nitration of the end products or starting materials of the process of the present invention promotes substitution at the 9 position, with minimal amounts of the 7-nitro derivatives sometimes being detected. To prepare 7-amino starting materials or 7-amino end products within the scope of the present invention, the best technique involves the preparation of the corresponding 7-azo aromatic derivative followed by reduction which gives the desired 7-amino derivative. , reduction preferably carried out by catalytic hydrolysis in the presence of a noble metal catalyst.

   In general, any diazotizable aromatic primary amine can be employed to prepare the 7-azo compounds, and it is essential that the

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 enol position 11, 12 is protected by an 11a-halogen group or any other suitable group which blocks. During the coupling reaction, it is preferable to use an excess of diazonium salt and to maintain the reaction temperature at 0-10 C.



   A mixture of aqueous solvents such as a 50% aqueous solution of a water soluble lower alcohol or a ketone may be suitable. t
Examples XLVI-XLVII illustrate the diazonium coupling reaction followed by reduction to obtain the 7-amino compound.



   The above description of the process of the present invention shows that the substitution on the D ring of the final product can be obtained from a judicious choice of the starting products substituted on the D ring or by a judicious substitution of the ring. D of selected 6-epi6-deoxytetracycline or 6-deoxytetracycline. In general, the best technique involves substitution; ' cycle D of the product resulting from the hydrogenation process claimed herein because the resulting yields are usually higher.



   In summarizing the process of the present invention, it will be appreciated that it not only provides a convenient means of producing new and useful 6-epi-6-deoxytetracyclines but also gives known 6-deoxytetracyclines. Although the latter compounds can be obtained by hydrogenation of a homologous tetracyclic antibiotic, i.e. containing both a 6-methyl and 6-hydroxy substituent, the process of the present invention is however. better because the yields are significantly higher than those obtained by the usual process.



   The hydrogenation of the 6-deoxy-6-demethyl-6-methylenetetracycline starting materials of the present invention, when carried out under the best acidic conditions, sometimes also produces a small amount of 4-epi derivatives. If desired, these 4-epi compounds can be separated from compounds having a normal 4-position configuration by conventional paper chromatography. In addition, the 4-desdimethylamino analogues of the pro

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 The starting materials of the present invention defined above can be used to produce the 4-desdimethylamino analogs of the products in question.



   The products of the present invention include 6-epi-6-deoxy-, 6-epi-6-deoxy-7-substituted-, 6-epi-6-deoxy-9substituted-, 6-epi-6-deoxy-11a -chloro-, 6-epi-6-deoxy-11a-fluoro-, 6-epi-6-deoxy-7, 11a-dichloro-, 6-epi-6-deoxy-7-substituted-9-substituted- , and the analogues lla-fluoro- and chloro- of the latter compound and 6-epi-6-deoxy-7-chloro-lla-fluorotetracyclines, with and without 5-hydroxyl- substituent, as well as their salts obtained by addition of diacid and their pharmaceutically acceptable metal salts, the substituents at positions 7 and 9 being selected from chloro, bromo, iodo, nitro, amino and lower alkylamino groups.

   Among these, one of the best groups of compounds includes those comprising a 5-hydroxyl substituent, since they have particularly good in vivo efficacy.



   The new compounds which have just been described, with the exception of the 9-nitro- compounds, are useful because of their high activity both in vivo and in vitro against a variety of microorganisms. The 9-nitro- compounds usually have lower activity. These nitro compounds are useful for the preparation of the corresponding 9-amino compounds as described herein.



   The table below summarizes the comparative in vitro activity of 6-epi-6-deoxy-5-oxytetracycline (1) and 6-deoxy-5-oxytetracycline hydrochloride (2) against a variety of destructive microorganisms. The minimum inhibitory concentration (MIC) is determined by the well known technique of serial dilution.

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  0i'Gf.? IIi14E * AIC (mc6fml) (1) (2) icrococcus pyogenes, var.aureus 0.78 0.78 Streptococcus pyogenes 0.09 0.09 Streptococcus fascalis 0.39 0.78 Diplococcus pneumoniae 0.19 (pi) 0.39 Erysipelothrix rhusiopathiae 0.39 0.39 Corynebacterium diphtheriae 0.19 1.56 Listeria monocytogenes 0.19 1.56 Bacillus subtilis 0.19 0.19
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 Bacterium awnoniagenes - 0.78 Aerobacter aerogenes 6.3 12.5 Escherichia coli 3.12 12.5 Proteus vulgaris 100> 200 Pseudononas aeruginosa 12.5> 200 Salmonella typhosa 6.3 6.3 Salmonella gallinarum 3.12 12.5 Klebsiella pneumoniae 3.16 6.3 Neisseria gonorrhoeae 0.09 6.3 Hemophilus influenzae 0.09 0.39 Shigella sonnei 3.12 6.3 Brucella bronchiseptica 0.78 3.12 Malleomyces mallei 0,

  78 25 Vibrio comma 0.39 6.3 Pasteurella multocida 0.19 Streptococcus agalactiae 0.19 -
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 Antibiotic resistant strains of Micrococcus pyogenes var. aureus: 376 6.3 (p.i.)> 200 400 6.3 (p.i.) 200 (p.i.) = partial inhibition.



   In the following table the properties are compared
 EMI11.4
 6-epi-6-d6soxY-5-oxy tetracycline (1) and 6-deoxY-5oxytetracycline (2): Properties of 6-deoxy-oxytetracYclines.



   (1) (2) Bioactivity against K. pneumoniage
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 (relative to ox.3rtetracTcline) Ca. 1500 y / mg Ca. 500 bzz 1 max HC1-jJ, ethanol 267 266 (= 17.300) (= 18.10Q)
351 343 (@ = 13. 200) (@ = 13.200) Rf @ 0.5 0.3
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 x Chromatography on paper saturated with aqueous buffer ¯4ac ilvain pH 3.5, developed for one hour with nitronethzne: chlorororm: pyridine 0: 10: 3 by volume.

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 EMI12.1
 



  The .10u.veaux C0 :: lpOS:; S de La ô-c? I-6-a6soxyt-.c / lL.iL-'i- ....... 1 --Y "! - .. Pf '{tf co-mùosi-.i-) ns .... ". 1' t'- piéciJ <-. pil = -. r .-. = r¯u ", 11- s in different CJ21JOS1. Ll'Wl :: .... <: - ¯0," ". to those of the homologous tetracyclines dorn. they are derived. sound: therapeutically useful in al inents or Cj: .. 't.
 EMI12.2
 growth stimulants, in veterinary practice and in agriculture.



  For therapy in men, the usual normal dose of these new compounds is from about 0.1 to about per day for an average adult. The product is formulated under the form of
 EMI12.3
 capsules or compri.nés containing 25 to 250 .1g of antibiotic on an activity basis. Suspensions and solutions in various vehicles are prepared using concentrations ranging from 5 to 125 mg ml. For intramuscular or intravenous parenteral administration, the daily dose is reduced to about 0.05 to 1.0 g. Intramuscular formulations include these solutions
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 of antibiotic at concentrations ≥1 from 50 to 100: nç / .al.

   Intravenous administration is done using isotonic solutions having an antibiotic concentration of about 10 mg ml.



  Both types of parenteral products are supplied as solid compositions for reconstitution. These products can also be used for applications on the usual growing media. In any case, the attending physician must
 EMI12.5
 Obviously, indicate the dose which is appropriate for each patient in particular. Smaller doses will be used for children.



   The present invention relates to all salts, including metal salts and those obtained by addition of acid, of newly recognized amphoteric antibiotics. The processes well
 EMI12.6
 known preparations of salts of tetracycliqa compounds are applicable in this case and are illustrated by examples below. These salts can be formed with metals and acids which are pharmaceutically acceptable, but also unacceptable. By "pharmaceutically acceptable" is meant metals and acids capable of forming salts which do not appreciably increase the toxicity of amphoteric antimirobial agents.

   The best salts are those obtained by adding acid and

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 pharmaceutically acceptable metal salts.



   The salts obtained by diacid addition, which are acceptable from a pharmaceutical point of view, are particularly advantageous from a therapeutic point of view. They include salts of mineral acids such as hydrochloric, hydriodic, hydrobromic, phosphoric, metaphosphoric, nitric and sulfuric acids, as well as salts of organic acids such as tartaric, acetic, citric, malic, benzoic acids. , glycolic, gluconic, gulonic, succinic, arylsulfonic, for example p-toluenesulfonic, and the like. Pharmaceutically unacceptable salts obtained by addition of acids, although they are not used therapeutically, can be employed for the isolation and purification of the recognized new antibiotic.

   In addition, they can be employed for the preparation of pharmaceutically acceptable salts. Of these, the most common include salts formed with hydrofluoric and perchloric acids. The hydrofluorides are particularly useful for the preparation of pharmaceutically acceptable salts, for example the hydrochloride, by solution in hydrochloric acid and crystallization of the hydrochloride obtained. The salts of perchloric acid are useful for the purification and crystallization of the new antibiotic.



   While all metal salts can be prepared and are employed for different purposes, pharmaceutically acceptable metal salts have particular value because of their therapeutic utility. Pharmaceutically acceptable metals most commonly include sodium, potassium and alkaline earth metals of atomic number less than or equal to 20, such as magnesium and calcium, and also aluminum, zinc, iron. and manganese, among others. Indeed, the metal salts include complex salts such as metal chelates, which are well known in the field of tetracycline.

   Metal salts unacceptable from the point of view

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 Pharmaceuticals most often include lithium salts and.: lethal alkaline earth metals with atomic number greater than 20, co.n.ne barium and strontium, which are used for isolation and purification, antibiotic . Because the new antibiotic is amphoteric, it also forms salts with sufficiently basic amino acids.



   It is evident that, in addition to their therapeutic qualities, the pharmaceutically acceptable acid and metal salts are also useful for isolation and purification.



   The following examples are given by way of indication and do not in any way limit the invention, many variations being possible within the scope thereof.



  EXAMPLE I. - 11a-fluoro-5-oxytetracycline-6,12-hemiketal.



   1 a mixture of 6.9 g of the anhydrous oxytetracycline base dissolved in 285: nl of methanol cooled in an ice bath, one equivalent of a 1N solution of sodium methoxide in methanol is added. The yellow sodium salt precipitates. A stream of perchloryl fluoride is passed which dissolves the sodium salt, when the mixture approaches neutral, a heavy precipitate begins to form. The excess perchoryl fluoride is removed with a stream of nitrogen, the product is filtered off, washed with cold methanol and dried under vacuum at room temperature to obtain 5.1 g of pale yellow crystals. Infrared absorption does not show carbonyl absorption below 6 microns.

   A bioassay on K. pneumoniae shows an activity of 4 meg mg on the tetracycline scale. Ultraviolet absorption shows maximums at 265 and 335 microns. Elemental analysis gives the following results after recrystallization of the product in water: Calculated for C22H23O9N2F.2H2O: C, 51.4; H, 5.25; N, 5.5 Found C, 51.2; H, 5.3; N, 5.7.

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 EMI15.1
 



  . "¯..i..¯ ': i'L .l.l.lla-cnloro-5-oxytetracycline-6,12-hemicetal.



  23 g of anhydrous oxytetracycline are dissolved in 250 μl of 1,2-imethoxyethane and then 34 N-chlorosuccinamide is added. The mixture is stirred for two minutes and then poured into 1 liter of water with stirring. The product which separates is collected by filtration, washed with water and dried. The infrared analysis of the product (KBr at a concentration of 1%) does not show any absorption due to carbonyl in the region of 5-6 microns but
 EMI15.2
 against the following main peaks: b, 12, 6.35, 6.66, 6.5, 7,; z, 7.55, 7.75, 7.92, 8.1..4., 8.36 , 8.76, 9.18, 9.43 microns, A bioassay against tetracycline activity of 4c /. .



  .: 0 # .IPLE III. lla-chloro-6-aéso -6-deeth 1-b-: nethvlane --- o tetrac cline.



  5 g of 1α-chloro-5-ox, tetracycline-b, 12-hei-ketal with 15 μl of liquid and dry hydrofluoric acid are added and the mixture is stirred for 3.5 hours at the temperature of the ice bath. . The hydrofluoric acid is evaporated off by heating under a stream of nitrogen to obtain the hydrofluoride.



   The crude hydrofluoride is dissolved in water and concentrated HCl or perchloric acid is added dropwise.
 EMI15.3
 (70;) to precipitate the hydrochloride or perchlorate. The hydrochloride is precipitated from an acetone solution of the crude hydrofluoride by addition of 47% hydriodic acid.



   Or, alternatively, the initial reaction mixture is ciluted with 6 or 7 volumes of water and ice and perchloric acid or naphthalenesulfonic acid (concentrated acid) is added to precipitate their respective salts. Acetone dilution of the initial reaction mixture, followed by addition of HI, precipitates the iohydrate.
 EMI15.4
 



  Elemental analysis of the hydrochloride gives the following values: Calculated for C22H21Ii20gCl1.FiI: C, 43.7; IM, 3.7; 11, 4.6; Cl, 5.8 Found C, 44.0; H, 4.0; N, 4.2; Cl, 5.5.

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  Ultraviolet analysis shows the following maxima: 222, 270 and 372 m. Infrared analysis shows the main peaks at 3,05,3,2, 5,7,
 EMI16.1
 6, Ô2, 6.03, 6.22, 6,., 6.88, 7.4, 7, $, 8.1, 8.9, and 9.1 microns. Perchlorate shows on ultraviolet analysis maxima at 237, 270 and 372 m.
 EMI16.2
 



  11a-fluoro-6-deoxy-6-demethyl-6-methylene-5-oxytetra-cyclin is prepared in the same way using lla-fluoro-5- 'o: tetracycline-b, 12-hemiaetal as the compound of departure.



  EXAMPLE IV - 6-deoxt-6-demethyl-6-methYlene-5-oxytetracycline.



  Method A.



   To a solution of 5 g of the product of Example III (under
 EMI16.3
 form of hydrochloride) in 125 ml of dilute hydrochloric acid (1 part of concentrated HCl in 55 parts of water) at 20 ° C., 2 g of powdered zinc are added. After stirring for 10 minutes, the zinc was removed by filtration, the filtrate was brought to a pH of 0.3 and extracted with butanol. The butanol extract is concentrated under reduced pressure until a residue is obtained which is triturated with ether. The residue insoluble in ether is crystallized from a mixture of methanol-acetone-HCl conc.-ether to obtain chlor-
 EMI16.4
 hydrate-aononethanolate (2.5 g) which melts and decomposes at 205 C.

   Ultraviolet analysis in 0.01N HCl in methanol shows the
 EMI16.5
 maxima 1 \ max 252 n / u, h '1 cm 450 and max 345 bzz' Ll '1 cm 302; in NaOR OOIN in methanol, 235 mu, ig., 442; in NAOH O'Olli in methanol, max 235 m / u, E 1 cm j42; ; 4u. 1 &.40g; At 385 m ,, U.J, 1 -: 1.329; max 2 'm / u E 1 cm, 408; max 38S m! u cm '39, ninf 2b0 m / u' El '1 cmj, 329; in 0.01N MgCl2 in methanol, At max 240 m / u, Eloem 1461; Max 277 n / u, E1 /, 1 cm 326; At max 351 m u, E1% cm, 282.



  Infrared analysis shows the main peaks at 6.03; 6.2; 6.37 and 6.87 microns. A bioassay shows a value of 2,000 to
 EMI16.6
 2,400 mcg / mg (turbimetric test on K. pneumoniae with reference to oxytetracyclinc). Elemental analysis of the product gives the following values: C, 55.0; H, 5.2; N, 5.5; Cl, 7.0; OCH3, 3.4.

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  The product shows Rf values of 0 and 0.35 respectively in the following systems:
Mobile phase Immobile phase (1) 20: 3 toluene-pyridine buffer pH 4.2 (aqueous) saturated with pH 4.2 buffer (2) 20: 10: 3 buffer pH 3.5 (aqueous) chloroform, pyridine saturated with a pH 3.5 buffer.



  Method B.



   A mixture of 1 g of the 11a-chloro product of Example III in 10 ml of methanol containing 200 mg of 5% rhodium on carbon is hydrogenated at room temperature and under a hydrogen atmosphere until that an equimolar amount of hydrogen is absorbed (2 hours). The catalyst is filtered off, the filtrate is evaporated to dryness and the residue crystallized as in method A.



  Method C.



   A mixture of 1 g of the 11a-chloro product of Example III in 70 ml of water containing 1 g of sodium hydrosulphite is stirred for 0.5 hour at room temperature. The mixture is then extracted with butanol and the butanol extract is evaporated to dryness.



  The product is crystallized from the retort residue in method A.



  Method B.



   Using the procedure of method A, we reduce
 EMI17.1
 11a-rluero-6-deoxy-6-denethyl-6-methylene-5-oxytetracycline to 6-deoxy-6-demethyl-6-methylene-5-oxytetracycline perchlorate.



   The crystalline hydrochloride-methanolate of this example can be recrystallized from isopropanol to give chlor-
 EMI17.2
 6-deoxy-b-demethyl-6-methylene-5-o-ry tetracycline hydrate. The recrystallized product shows the following peaks on infrared analysis: 3,1,3,75, 6,02, 6,23, 6,36,6,55, 6,9, 7,35, 7,7,7 , 8, 8.15, 8.26, 8.5, 9.27, 9.95, 10.55, 10.8, 11.53, 11.93 and 12.15 microns.

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 EXAMPLE V.-
 EMI18.1
 7, 11a-dichlojo-6-deoxv-6-denethyl-6-methYlene-5.-oxYtetracvcline.



  Method A.



   1.5 g of N-chlorosuccinimide are added to 5 g of hydrochloride
 EMI18.2
 1a-chloro-d-deoxy-6-demethyl-6-methylene-5-oxytetracycline in 15 ml of liquid hydrofluoric acid at the temperature of the ice bath. The solution is stirred at the temperature of the ice bath for 1.5 hours. The crude product is precipitated by the addition of 500 ml of ether and collected by filtration.



   The crude product is taken up in methanol at room temperature, the insoluble product is filtered off, the filtrate is treated with reactive carbon, filtered and concentrated under reduced pressure. The remaining residue is taken up in dilute hydrochloric acid from which the product is recrystallized as hydrochloride.



  Ultraviolet analysis in 0.01N HCl in methanol shows
 EMI18.3
 Àmax239 mu 1 m 352, À max 376 mu El% cm 1,60, \ ion z5g mou E1% cm 324. I.R. analysis shows the main bands at 5.7, 6.0 and 5.9 / u.



  Method B.-
5 g of 11a-chloro-5-oxytetracycline-6,12-hemiketal are added to 15 ml of liquid hydrofluoric acid at ice bath temperature. After having been stirred for 3.5 hours at the temperature of the ice bath, method A is applied after addition of the same weight of N-chlorosuccinimide to obtain the product.



   Another slightly better method is the following. After having removed most of the liquid hydrofluoric acid, 100 ml of water are added and then 5 g of -naphthalenesulfonic acid.



  The product precipitates as -naphthalenesulfonate and is collected by filtration.



   Another method involves diluting the initial reaction mixture in 6-7 volumes of water, followed by the dropwise addition of the concentrated acid to precipitate the perchlorate and the -
 EMI18.4
 retort naphthalenesulfonzte it is described in example III. The per-

 <Desc / Clms Page number 19>

 The crude chlorate thus obtained is crystallized in the form of long needles from isopropanol and shows on ultraviolet analysis maxima at 260 and 377 m / u and an inflection at 260 m. Infrared analysis shows Dics at 5.7, 6.0, 6.26, 6.55, 6.88, 7.2, 7.85 and 8.35 microns.



    EXAMPLE VI.-
 EMI19.1
 ? -chloro-b-deox-b-demethyl-b-methYlene-5-oxytetracycl ne.



  Method A.



   0.45 g of sodium hydrosulfite is added to a solution
 EMI19.2
 0.5 g of 7, lla-dichloro-6-deoxy-6-dexethyl-6methylene-5-oxytetracyclin.e perchlorate in 7.5 ml of water and the resulting mixture is stirred for 12 minutes. The product separates and is collected by filtration. A biological test of the product gives a value of 3,400 mcg mg (turbimetric test on K. pneumoniae with reference to
 EMI19.3
 with oxytetracycline (1000 mcg / mg)).



  Method B.



  20 g of -rphthalenesulphonate from the preceding example are suspended in 500 ml of methanol containing 5 g of 5% rhodium on carbon and the mixture is hydrogenated at room temperature under 1 atmosphere of hydrogen. When 700 ml of hydrogen had been absorbed, the reaction mixture was filtered and the filtrate evaporated to dryness to obtain 15.4 g of residue.



   The methanolic solution of 11 g of the residue is then adjusted to a pH of 6.5 with triethylamine and passed through an 8 x 100 cm column containing 2 kg of cellulose powder, water being the fixed phase. The column is eluted with water-saturated ethyl acetate and 45 ml fractions are collected. The progress of the elution is followed by chromatography on paper and the fractions 132 to 260 are collected, evaporated to dryness, stirred in ether and filtered to give 2.74 g of a pure and amorphous amphoteric product.



   The product is crystallized by dissolving 1.6 g in 40 ml of hot methanol and scraping off. Filtered and collected 890 mg of

 <Desc / Clms Page number 20>

 
 EMI20.1
 produce in basic form & -: 1;>: - 10 t ::: re. J ..: '(,. :: é; .l :: e infrared 1 <: OE'i ± the following peaks: 2.9C, 3.29, 3.4z, 6.06, 6.1, 6, 30,, 5 ?, -, 7.19, 7.4, 7.70, 3, ij, 9.06, 9.33, 1063, .. 0.92, 11.55 and 11.76 '; 1crons .

   Anal; is ultraviolet against: in 15Cl 0.011; in methanol, des: na :: i: 1 & at 247: '1 / u (106 t: 4.2 <D) and 346 ID / u (lodged 4, J2) and an inflection at 370 -. / -1 u (batch 1 3.9; 3); in O, OlN in methanol, these :: Iéxiuê. at 23 ./u (logE4.24), 253: a / u (log E. 4.22) and 389: au (log 4.12) and an inflection at 284: "/ 1.1 (log E. 4.07 ); in: 3Cl .. :: 301.-! in the '1 {té.nol, (es Jl.xi.ll6. to 241 :: 1 / 1.1 (logg4,32), 349 mu (low <4, 04) and 372 a / 1i (epauleaent) {loË,., 02).



   The product against the following Hf values in solvent systems indicated:
 EMI20.2
 Dc value Lf Phase! Lobile 2ystene c'e solvents Phase iJ1TIobilE.



  O, JS ethyl acetate aqueous phosphate buffer saturated with e4u JE 3, J, 33 ethyl acetate tcuapon de: 1etc Ilvaine saturates G'ea "01 pH 4, Un ess¯1 tiologicue (on K. PEE: u : 10nié ... e en se n.9;: ort &n; - oY'tétré :: .- cycline) conne a value of 6000 mcg / mg.
 EMI20.3
 The crystalline perchlorate of the product of the previous example is hydrogenated to give this product which crystallizes out. from methanol and perchloric acid at 70.,. The perchlorate of the product against an ultraviolet spectrum identical to that of the amphoteric base.



  Method C.



   This product can also be obtained by reduction of the starting compound of the process ± with zinc and an acid according to the process of example IV.



  EXAMPLE VII. -
 EMI20.4
 7-bro yo-lla-chloro-6-deox; y-6-demethvl-6-: uc thylene-5-oxytetrac rcline.



  This product is prepared according to processes A and B of
 EMI20.5
 1. "Example; lple V using an equivalent amount of α-broL-1osuccinimide instead of N-chlorosucciniraide.

 <Desc / Clms Page number 21>

 EXAMPLE VIII. -
 EMI21.1
 7-br: o - d soxa -, - dc: thYl-6 -,:, ë tb: l Donkey - o-r. tr trr c; cl in c.



  This product is obtained from the product of 1-E, -. ,, - ,, - LVII by treating it with the short-term hydrosulphite described in Example VI.



   The following compounds are prepared from the compounds
 EMI21.2
 1? A-halogenated by the process of e: ē.ples proceed ?: 7-chloro-lla-: luoro-6-deoxy-6-demethyl-6-: aé-thylene-5-oz.y tetrüc, cline '; -iodo-11G-chloro-6-deoxy-6-demethyl-6-: ct:; lene-5-o:., té trac; clīe 7-broo-lla-fluoro-é-désor-6-dénéth ; l - aEtbylene-5-oryt4 trac;, cline.



   These compounds are converted to their corresponding 11a-dehalogenated derivatives by the above-mentioned methods.



  EXAMPLE IX. -
 EMI21.3
 7 -cz: lo ro-9-ni tro-6-d é sox.y-6-derné thYl-ô-mé th = Tléne-5-o y té trn c¯, cl in e.



  Has a solution of 1.0 g of the base 7-chloro-6-cesox, -6- to..et¯ßl-6-methylene-5-oxtetrac3cline in: 3.: Il diacid fluorh; anhydrous liquid cricuc (3F ) at 0 C, add 220 .n #; de Le: .1f: l &r:; e is stirred for 30 minutes at 0 C and HF is removed with a stream of nitrogen. The residue is triturated in dry ether, filtered and dried.



  Below are the values of Rf in the indicated solvent systems: Rf Solvent system-
Mobile phase Immobile phase 0.2 ethyl acetate aqueous phosphate buffer saturated with water (pH on p 0.55 nitromethane, Mac Ilvaine buffer toluene, pyridine (pH = 3.5) (20: 10: 3) L ' ultraviolet analysis in 0.01 M NaOH in methanol against maxima at 248, 341 and 447 m / u and a shoulder at 275 m. A bioassay shows a value less than 100 mcg / mg (turbimetric test on K. pneumoniae in comparison with oxytetracycline).



  EXAMPLE X. -
 EMI21.4
 9-nitro-7,11a-dichloro-6-deoxv-6-demethyl-6-methylene-5-oxytetracvc7 To a solution of 600 mg of 7, lla-di- perchlorate

 <Desc / Clms Page number 22>

 
 EMI22.1
 chloI'OO-deoxy-o-èé.:. tetY.y: -o-.n.jt: -: Jl;:; ne-5-o ;; Q t.trilcycl in in .1.,: .. :: 1 decides liquid hydrofluoric ûnīy: .rc, 100 'Olt is added. de .r: t03 "The mixture is stirred at 5 ° C for 15 minutes, and the product is then precipitated with ether and collected by filtration, after washing with ether.
 EMI22.2
 



  EXE.:.1PLE I. - 9-nitro-6-deoxv-6-derethyl-6-ethylene-5-oxxtetr4c-cli. ^. E.



  Has a mixture of 500 ng of 6-deoxy-6-deEiethyl-6-methylene-5-oxytetracycline in 1.5 ml of anhydrous hydrofluoric acid and
 EMI22.3
 liçuide, 100 mg of SN03 are added. The mixture is stirred at 15 ° C for 30 minutes and then evaporated to dryness. The residue is triturated in ether and filtered to obtain the crude product which is triturated.
 EMI22.4
 in 5ro aqueous HCl and extracted with butanol. The butanolic extract is concentrated day to obtain the product. EXAMPLE XII.-
 EMI22.5
 nitro-11a-chloro-6-deso; g-6-ddaethyl-à-.aethylene-5 -> atétr-c; .cline.



  A sulution of 5 of 11a-chloro-3oxytetracycl-in-6,12-he-ketal in 10 inl of anhydrous fluoIllycric acid is allowed to stand for 3 l / h. then add 1.2 g of KNO3 to it while stirring. Stirring is continued for 1 hour at the temperature of the ice bath and the mixture is then poured into 200 ml of dry ether, filtered and washed with ether to obtain the crude product.
 EMI22.6
 



  EXE. "IPLE XIZI. - 7-chloro-9-0.nino-6-deoxy-6-demethyl-6-methylene-5-oxytetracycline. H a solution of 1.42 g of 7-chloro-9-nitro- 6-deoxy-6demethyl-6-methylene-5-oxytetracycline in 50 µl of water and 2 µl of cane HCl. 1.3 g of powdered zinc are added to 20 with stirring.



  After 15 minutes the mixture is filtered, the pH adjusted to 2.5 and the mixture is extracted with butanol (5 x 30 µl). The butanolic extract gives, after concentration, 760 mg of the product (in the form of
 EMI22.7
 hydrochloride). A bioassay (K.pneuraoniae) gives a value of 1680 mcg / mg. Ultraviolet analysis in 0.01 N HCl in MeOH
 EMI22.8
 shows maxima at 262 and 348 m / u; in 0.01A NaOH; .0E, at 264 and

 <Desc / Clms Page number 23>

 
 EMI23.1
 350 L I u.



  The product gives the values of Pffuivants:
Rf Solvent system
Mobile phase Immobile phase
0.2 ethyl acetate aqueous buffer saturated with water (pH = 3)
 EMI23.2
 0.35 nitronethane, Me Ilvaine buffer chloroform, pyridine (pH = 3.5) (20: 10: 3) This product can also be obtained by reduction (with
 EMI23.3
 aqueous sodium hydrosulfide) of 9-ritro-7, 11a-chloro-6deoxy-6-demethyl-6-methylene-5-oxyte tracycline.



  EXAMPLE XIV - 9-a.ino-é-de soxy - defe th vl-d-thylene-5-oxytetr4ccline.



   This product is obtained by reduction of the 9-nitrocorresponding derivative with sodium hydrosulfite in water or SnCl2 in this aqueous hydrochloric acid.



    EXAMPLE XV.-
 EMI23.4
  -nitro-11a-chloro-6-deo; v-to-xethyl-6-methylene-5-oxytetracycline. r a solution of 1 g of lla-chloro-6-deoxy-6-de'nëthyl-6-: ethylene-5-oxytetracyclire in 20: nl diacetic acid, 1 ml of concentrated nitric acid is added. The mixture is left to stand for 2 hours, then evaporated to a quarter of its initial volume and 200 ml of ether are then added. The product separates out as nitra. te and is collected by filtration.



   The corresponding IIa-fluorocortical derivative is prepared in the same way.



  EXAMPLE XVI. -
 EMI23.5
 -aino-6-deoxsr-6-dëétnvl-6-néthvlene-5-oxytetracycline.



   This product is obtained by reducing the product of the preceding example according to the process of example IV.



   This product can also be obtained by chemical reduction
 EMI23.6
 or catalytic 9-nitro-6-esoxy-6-emethyl-6-metiaylene-5-oxytetracycline.

 <Desc / Clms Page number 24>

 EXAMPLE XVII. -
 EMI24.1
 7 lla-dichloro- -n: itro-6-desc -6-demethT 1-6-meth lene- -ō tee: tra-cyclin. cycline. 1 g of 7,11a-dietdoro-6-deozy-6-demethyl-6-methyl-5-OxYtetracycline hydrochloride is dissolved in 20 ml of acetic acid and 1 ml of conc. HNO3 is added thereto. mixture is left to stand for 12 hours and then is concentrated to a quarter of its initial volume. 200 ml of ether are added dropwise thereto with stirring and while maintaining in an ice bath; stirring is continued for 3 hours and then the solid is collected by filtration.

   The solid is triturated several times in ether and then dried, and the product is thus obtained in the form of the hydrochloride.



   Using the same process, the following products were obtained from the corresponding initial derivatives:
 EMI24.2
 7-chloro-lla-fluoro-9-nitro-6-deoxy-6-demethyl-6-methylene-5oxytétracycline 7-brono-lla-chloro-9-nitro-6-deox3r-6-de.nétiayl-6-methylene -5-oxy.tetracycline 7-iodo-11a-fluoro-9-nitro-6-esoxy-6-denmethyl-6-methylene-5-oxytetracyline.



   These products are converted to 7-broino-, 7-iodo- and 7-
 EMI24.3
 chloro-9-amino-6-deoxy-6-demethyl-6-ethylene-5-oytetracycline by the reduction methods of the preceding examples. EXAMPLE XVIII.-
 EMI24.4
 -form lamino-6-deso. -6-demeth 1-6-aethene- -o tetrac cline A solution of 4.5 g of 1; hase 9-amino-6-deoxy-6demetal-6-; ethylene-5-oxytetracycline in 40 ml of dry pyridine, 20 ml of acetylformic acid are added at the temperature of the ice bath. The mixture is stirred for 15 minutes at ice cold temperature and then 1 liter of dry ether is added dropwise. The final product is filtered, resuspended in ether and refiltered.



   The crude product is suspended in 100 ml of water

 <Desc / Clms Page number 25>

 producing a solution of pH: 4.2 and stirred for 10 minutes in order to hydrolyze the 0-formyl groups. The pH of the mixture is then adjusted to pH = 5.5 and the solution is dried by freezing it, which gives the crude product which is then chromatographed on a cellulose column so as to obtain the purified product in the main fraction.



    EXAMPLE XIX.-
 EMI25.1
 9-aeétvla.nino-6-deoxy-6-demethvl-6-methse-5-oxytetraeycline
To a solution of 4.2 g of the starting material of Example XVII in 200 ml of dry tetrahydrofuran and 200 ml of dry methanol is added 20 ml of acetic anhydride and the mixture is stirred for 45 minutes at room temperature . Most of the solvent is then removed and the mixture is poured dropwise into dry ether, filtered and the solid magma is re-titrated in ether, filtered and dried to give the crude product.



   The crude product is chromatographed as in Example XVIII and the main fraction gives the purified product.



   Using the methods of Examples XVIII and XIX, the following products were prepared:
 EMI25.2
 9-formylamino-7-chloro-6-deoxy-6-demethyl-6-methylene-5-oxytetracyline 9-acetalamino-7-chlaro-6-deoxy-6-detethyl-6-methylene-5-axytEtraculin 9-butyrylamino- 7-chloro-6-deoxy-6-demethyl-6-methylene-5-oxytetracycline 9-propionylanino-6-deoxy-6-demethyl-6-methylene-5-oxytetracycline 9-valerylamino-6-deoxy-6-demethyl- 6-methylene-5-oxytetracycline.



  EXAMPLE XX. - lla-chlorotetracycline-6,12-hemiketal.



   This compound is prepared according to the process of Example II by substituting the oxytetracycline of Example II with an equivalent amount of tetracycline.

 <Desc / Clms Page number 26>

   : EXAMPLE XXI.-
 EMI26.1
 .7, 11a-dichlorotetracycline-6,12-hemiketal.



   This compound esc prepared according to the method of Example XX employing an equivalent amount of tetracycline and replacing the water with ether or an ether-hexane mixture to precipitate the product.



  EXAMPLE XXII.-
 EMI26.2
 11a-chloro-6-deoxy-6-demethYl-6-meth, lenettracycline.



   The lla-chlorotetracycline-6,12-hemiketal is dissolved in liquid hydrofluoric acid (at a rate of 2 g / 15 ml) at 0 C. The mixture is maintained at this temperature for 10-15 minutes, then the acid hydrofluoric is driven off. The residue is triturated with ether and a solid product, 11a-chloro-6-deoxy-6demethyl-6-methylenetetracycline, is obtained in the form of its hydrochloride which is recrystallized from methanol.



   Alternatively, the crude hydrofluoride (10 g) is dissolved in 350 ml of water with heating and stirring. An equal volume of concentrated hydrochloric acid is added to the clear solution and the product crystallizes in the form of its hydrochloride. Elemental analysis of the hydrochloride thus obtained gives the following results:
 EMI26.3
 Calculated CEzONzCi: C, 53.11; He 4.56; N, 5.63; chloride 7.13 Found C, 52.62; H, 4.63; N, 5.54; chloride 6.84.



  Infrared analysis of the product as hydrochloride in a KBr pellet at a concentration of 1% shows absorption due to carbonyl at 5.70 as well as the following significant peaks: 6.1, 6.2 ", 6, 36, 6.45 (shoulder), 6.91, 7.85, 8.14, 8.55, 10.22,
 EMI26.4
 10, 55 and 10, 9 A bioassay of the product (K. pneumoniae) shows an oxytetracycline activity of 50-100 mcg / mg. Ultra- analysis
 EMI26.5
 sample violet in 0.01 # 1 HCl-methano1 shows maxima at 376, 278 and 242 m / u.

   The product has an Rf value of 0.2 to 0.3 in the following system: Mobile phase Immobile phase 20: 3 toluene-pyridine aqueous buffer at pH 4, saturated with buffer at pH 4.2

 <Desc / Clms Page number 27>

 By examining the chromatogram in ultraviolet light, we notice that the product stain is not very fluorescent. However, when spraying aqueous sodium hydrosulfite on it, it becomes strongly fluorescent.



     The C.4 epimer of this product, present in the impurity state, shows an Rf = 0.1 in this system.



  EXAMPLE XXIII. -
 EMI27.1
 ia-fluoro-6-dôsoxz-6-àémethyl-6-zéthyienetitr c cline. ¯ 250 ag of 11a-fluoro-tet-tracycline-6,12-henicëtal is stirred in 2 ml of 63% aqueous perchloric acid. The solid dissolves on heating at 60-65 C for 15 minutes, after which the mixture is cooled and water is added to it and 11a-fluoro-6- is obtained.
 EMI27.2
 deoxy-6-dexiethyl-6-a & thylenetetracycline in the form of its hydrochloride. Ultraviolet analysis reveals absorption similar to that of Example XXII.
 EMI27.3
 



  XE: .fPL r.:CT V. The compounds below are prepared starting from the corresponding llah.?oté',râcyclines-b,12-héicétGls, the lla-fluoro compounds by the process of Example XXIII and the other 1-lahalogenated compounds by the process of example XXII:
 EMI27.4
 7, lla-dichloro-b-deoxy-6-d ¯ é t'¯.; 1-'v-é tb.rlènetetracrcline 7-chloro-lla-fluoro-6-deoxy-6-deJethyl-b-2 (thylenetetracycline 1-Bromo-Ila-chloro-6-desor-6-né.nétnyl-6-r ^ et: lylenetetrücycline 7-iodo-lla-chloro-6-dGsoxy-6-de3Lethyl-6- "iethylenetetracycline EJ #. ' iPLE XXV.- 6-dnsot-6-de¯nethyl-b-methylenetetracycline.



  Method A.



   The product of Example XXII (5 mg) is dissolved in 3 ml of methanol and a freshly prepared solution of sodium hydrosulfite (20 mg in 2 ml of water) is added thereto. The mixture is allowed to stand for 15 minutes at room temperature, after which it is freed from methanol and extracted with butanol.

 <Desc / Clms Page number 28>

 
 EMI28.1
 



  The extract:. 'I ï;: lJl7.i:, lt. is CO: 1C: ¯n I '', ¯¯our get 1 .1 '.) c' .. iit .. 7: - -C¯¯ ''. ߯'1T1 -: àt: t: jil ¯ïlP.t "irF..C;, rClî: l '. L: ¯rC" .it "v C¯;, c.llL¯ from. ^ ¯'2'¯I under 1' <; '.. ie this JJ-tOl11 - ': Í..Á-L:! 1é.t. by ic <ôiliJ = i <;', <1 .. p-toluënesulphonic. Or, it is crystallized 2: from .-, iu under forze ée chloyàrte & r 4ûci tion -11-z-Cl concentrated on a test on 1 ±, piùewioniac against naked the products of 0'o.xyttra.c.'Í activity 0'o.xyttra.c.'Ícline d-au-, oins 11'jO. 1Cs / lg.

   The y3Z'O i.; 1 tsp. ; 3.1's = hf value of 36 in the syst:;.; 1 ", described in 1'c: CF.; LE 1.Ziii r a value r, r (: e 8.9 in the following S7St; l8 : Phase; labilE: t'flL, S 1: 1.mobile? Liti, o.Jé.it; ini-chloroJfor, Je- t <, -: pon r¯c; ux 3> 5 pyridine 1 (): 3 L'0 .. :: Ji: "1? 1 ', ::'.: We C. t ;. of this substance z = es: 1 <.. 1: .C" 'mf d' -: - 1l '.'L -') 2: 1, L, and C'CfiVlrC? 1 06 oin both systems: the.
 EMI28.2
 



  Method
 EMI28.3
 , '18 .'1oc: e We refroioit a. 4C in 6th lbce ur bath ,. "option: 1 5 ') b of hydrochloride ce i14-chlor.-b-c%. :: r - :. ¯.t: ¯;' -.



  ; .1t710ne.; Trèc7cline in 500: il à 'é t' = er ... on3-ithyli:, uc or ct'u'-i en Clycol and this stirred solution neiC; 1J.s.ent is treated with 30 g due to the added zinc; Z '& Ci, lC.l.lE' '? E' = in 10 'îl.¯ "I', lvë approximately. La t - ::::. 1, t; sré :. Lure rises to 12 C during this addition.



  When the addition is finished, the temperature C () J'1 ("nce to drop." After a total reaction time of 15 minutes, the zinc
 EMI28.4
 is removed by rapid filtration and washed with a solvent. We
 EMI28.5
 add 1 liter of E: .é: .1.1 to the filtrate, gradually over about 10 minutes. The complex of the product with zinc forms a yellow sludge.



  The pl of the solution is then adjusted to 6.8 with 10% aqueous sodium hydroxide.

 <Desc / Clms Page number 29>

 



   The resulting slurry is allowed to stand for about 1 1/2 hours in an ice bath and filtered. The wet cake is then triturated in 750 ml of water and concentrated HCl is added dropwise until a clear solution is obtained. A slight excess of concentrated HCl causes rapid crystallization of 6-deoxy-6-demethyl-6-methyleneetetracycline hydrochloride in the form of shiny needles. After standing for one hour, the product is filtered and dried. The yield is 37.8 g.



  The product melts and decomposes at 213.8-214.2 C.



  EXAMPLE XXVI. -
The process of Example XXV is repeated to dechlorinate the 11a-chloro-6-methylenetetracycline from the previous examples, to obtain the following products:
 EMI29.1
 7-Bromo-6-deoxy-6-demethyl-6-methylenetetracycline 7-chloro-f-deoxy-b-demethyl-E-methylenetetracycline C.4 epimers can appear as impurities. EXAMPLE XXVII, -
 EMI29.2
 lla-chloro- -nitro-b-deso -6-demé h 1-6-mpth lénetétr c c ne
A mixture of 1 g of the product of Example XXII in 20 ml of acetic acid containing 1 ml of concentrated HNO3 is left to stand for 12 hours, after which the addition of 20 ml of H2O crystallizes the product in the form of nitrate.

   The product is collected by filtration, washed with water and dried.
 EMI29.3
 



  EXJ24PLE XKVI I I. - -nitro-6-dice 1-6-meth enetetrac cline.
This product is obtained from the corresponding 11a-chloro compound (Example XXVII) by treating 1 g in 70 ml of water with one equivalent of sodium hydrosulphite. The reaction mixture is stirred at room temperature for 1/2 hour and the desired product is obtained by butanol extraction and evaporation of the extract to dryness under reduced pressure.



  EXAMPLE XXIX.-
 EMI29.4
 7-chlçro-4-r¯i tro-6-deoxy-E-demé thZrh6-reth ,, lene té raçyçlin

 <Desc / Clms Page number 30>

 220 mg of KNO3 are added to a solution of 1.0 g of
 EMI30.1
 7-cbacro-6-desaxy-6-demethyl-6-methylenetetracycline in 8 ml of anhydrous liquid hydrofluoric acid (HF) at 19 C. The mixture is stirred for 30 minutes at 0 C and the HF evaporated under a stream of nitrogen. The residue is triturated in dry ether, filtered and dried.



  EXAMPLE XXX. -
 EMI30.2
 7-chloro-9-nmino-6-deoxv-6-demethyl-6-methylethetetracycline
1.3 g of zinc powder are added with stirring at 20 ° C. to a solution of 1.42 g of 7-chloro-9-nitro-6-descxy-6-demethyl-6-methylene-tetracycline dissolved in 50 ml. of water and 2 ml of concentrated @ Cl. After 15 minutes, the mixture is filtered, the pH is adjusted to 2.5 and the mixture is extracted with five times 30 ml of butanol. Concentration of the butanol extract gives the desired product as a dihydrochloride.



  EXAMPLE XXXI. -
 EMI30.3
 7lla-dichloro-9-r.itro-6-deoxy-6-demel-6-métthylènetétracvcline 1 g of 7, lla-dichloro-6-deoxy-6-demethyl-6-r.: Ethylenetetracyclin hydrochloride is dissolved. : in 20 ml of acetic acid and 1 ml of concentrated nitric acid is added. The mixture is allowed to stand for 12 hours and then concentrated to 1/4 of the original volume. It is added dropwise @ 200 ml of ether with stirring at ice bath temperature. Stirring is continued for 3 hours and the solid is collected by filtration. The solid is triturated several times in ether and then dried to obtain the product in the form of the hydrochloride.



   Using the same process the following products are obtained from the corresponding starting compounds: 7-chloro-lla-fluoro-9-nitro-6-deoxy-6-demethyl-6-methylenetetracycline 7-bromo-lla- chloro-9-nitro-6-deoxy-6-demethyl-6-methylenetetracycline
 EMI30.4
 7-iodo-Ila-fluoro-9-nitro-6-deoxy-6-denethyl-6-retethylenetetracy- cline

 <Desc / Clms Page number 31>

 These products are converted to 7-bromo-, 7-iodo- and 7-
 EMI31.1
 chloro-9-amino-6-fiesoxy-6-demethyl-6-methylenetetracycline by the reduction methods of the preceding examples. EXAMPLE XXXII.-
 EMI31.2
 z0 g of 6-deoxy-6-demethyl-6-ethylene-5-oxytetracycline, 5.1% of 0.01N HCl and 40 g of rhodium on 5p carbon are introduced into a common hydrogenation apparatus.

   Hydrogen is then introduced into the apparatus and maintained at a pressure of 1800 pounds / Soft square (126 kg / cm2) at a temperature of 30 C for 20 hours. Then the catalyst is filtered off and the filtrate is frozen and dried to obtain 23.9 g of solid, a mixture of 6-
 EMI31.3
 deoxy-5-cxytetracycline and 6-epi-6-deoxy-5-cxrtetracycline. The efficiency of this operation, amounting to about 60%, can be improved by employing 5% rhodium on a barium sulphate support in order to reduce absorption losses on the supporting catalyic carbon.



   A mixture of methanol and water (25 ml each) is used to dissolve 20.8 g of the crude solids from the previous operation. A solution of 20 g of sulfesalycilic acid in a mixture of methanol and water (50 ml of each) is added thereto. The
 EMI31.4
 6-desexy-5-oxSTtetracyclines readily crystallize from sulfosalycilates. The slurry is then filtered and the cake is washed with 50% aqueous methanol and dried to obtain 242 g of salt (operation yield 75%).



   Counter-current distribution is used to separate
 EMI31.5
 6-epi- & ohsoxy-5-oxytetracycline of 6-deoxy-5-oxytetracycline.



  For this purpose, 500 mg of the salt from the previous operation are dissolved in 30 ml of ethyl acetate plus 10 ml of 2M sodium acid phosphate. The two resulting phases are stirred and the pH adjusted to 6.8 by addition of Na2HPO4. The aqueous phase is separated in a separatory funnel and equilibrated with 30 ml of fresh ethyl acetate. This operation is repeated twice more with fresh portions of ethyl acetate and the aqueous phase is then put to

 <Desc / Clms Page number 32>

 
 EMI32.1
 side. Na2HPO4-KHZPO4 aqueous buffer pH 6.8 (10 ml) is equilibrated with each of the ethyl acetate layers in four vials and then set aside. This operation is repeated twice with a fresh buffer of pH 6.8.

   The four layers of ethyl acetate are combined and evaporated to dryness to give 172 mg of substantially pure 6-epi-6-deoxy-5-oxytetracycline (operation yield about 50%). The product remaining in the aqueous buffer layers is concentrated to give 6-deoxy-5, oxytetracycline (operation yield about 50%).
 EMI32.2
 



  The 6-epi-6-deoay-5-oxytetrayline from the previous operation is finally purified by dissolving it in 1 ml of ethanol and adding 3 drops of concentrated HC1 thereto. It crystallizes in the form of pure hydrochloride, the analysis of which is:
Found: C, 53.69; H, 5.81
Calculated: C, 53.86; H, 5.70
This product tested against oxytetracycline on K. pneumoniae shows 1400 units per milligram.



  EXAMPLE XXXIII.-
Hydrogen is introduced into a normal hydrogenation apparatus containing 10 g of 6-deoxy-6-demethyl- hydrochloride.
 EMI32.3
 6-methylene-5-azcytetracycline, 150 ml of methanol and 5 g of 5% rhodium on carbon. The pressure is maintained at 50 pounds / square inch (3.5 kg / cm2) with stirring at room temperature for 24 hours. The catalyst is then filtered; the cake washed with methanol and the combined filtrates are evaporated to dryness. The dried solids are triturated in ether, filtered and dried again. The resulting solids have a biological activity of 1345 units per milligram against K. pneumoniae.



   35 ml of water are used to dissolve 8.5 g of the above product and the pH is adjusted to 6 with triethylamine,
 EMI32.4
 adding enough diillethylformamide to hold the solids in solution. 2 kg of cellulose powder are suspended in water saturated with ethyl acetate and the whole is poured into a column about 9 cm in diameter and with a height of

 <Desc / Clms Page number 33>

 0.9 m. The product solution is then chromatographed on this column, eluting with approximately 12 liters of water saturated with ethyl acetate. The first fraction containing product collected under the column provides 1.85 g of 6-epi-6-deoxy-5-oxytetracycline. The following fraction contains 2.0g of 6-deoxy-
 EMI33.1
 6-demetbyl-6-methylene-5-ozytetracycline.

   The third fraction gives z8 g of 6-deoxy-5-oxytetracyclme.



  EXAMPLE XXXIV.-
Has a solution of 500 mg of 7-chloro-hydrochloride
 EMI33.2
 6-deoxy-6-demethyl-6-methylèhe-5-oxytetracycline in 25 ml of methanol, 500 mg of 5% palladium on carbon are added. The suspension is hydrogenated at atmospheric pressure until there is no more absorption of hydrogen. The catalyst is separated by filtration and the filtrate is evaporated to dryness in vacuo. Paper chromatography of the residue reveals high concentrations of 6-
 EMI33.3
 epi-6-deoxy-5-oxytetracycline and 6-deoxy-5-OxYtetracycline with traces of starting material.



   The process of the previous paragraph is repeated using as starting product the hydrochlorides of the following products:
 EMI33.4
 7-bromo-6-deoxy-6-demethyl-6-methylene-5-oxytetracycline 7-iodo-6-deoxy-6-demethyl-6-methylene-5-oxytetracycline 7-chloro-6-deoxy-6-demethyl- 6-methylene-5-oxytetracycline to obtain separable mixtures of the corresponding 7-dehalo-6-epi-6-deoxytetracyclines and 7-dehalo-6-deoxytetracycline.



   In the same way, the hydrochlorides of 7-chloro-
 EMI33.5
 9-amino-6-deoxy-6-demethyl-6-methylene-5-oxytetracycline, 7-chloro-9-acetylamino-6-esoy-6-demethyl-6-methylene-5-oxytetracyclme and 7chloro-9-amino- 6-deoxy-6-demethyl-6-methylenetetracycline to obtain separable mixtures of 7-dehalo-9-substituted-6-epi-6-deoxytetracyclines and 7-dehalo-9-substituted-6-deoxytetracyclines.



  Likewise, the hydrochlorides of? Lla-dichloro-9-ni = tro-6-deoxy-6-demethyl-6-methylenetetracycline and 7-bromo-lla-chloro-9-nitro-6-deoxy-6- demethyl-6-methylenetetracycline are reduced to separable mixtures of 9-amino-6-epi-deoxytetracycline and 9-amino-6-deoxytetracycline.

 <Desc / Clms Page number 34>

 



  EXAMPLE XXXV.-
To a solution of 500 mg of 7-chloro-6-deoxy-6-d - methyl-6-methYlene-5-oxytitracycline hydrochloride in 25 ml of methanol, 500 mg of rhodium on carbon 5% are added. The suspension is hydrogenated at a pressure of 2,000 p.s.i. until one mole of hydrogen is absorbed. The catalyst is removed by filtration and the filtrate evaporated to dryness in vacuo. Paper chromatography using the ethyl acetate-water solvent system is employed to isolate 7-chloro-6-epi-6-deoxy-5-oxytetracycline.



  EXAMPLE XXXVI.-
A suspension of 1 g of platinum dioxide in 500 ml of methanol is saturated with hydrogen. We add a 15g solution
 EMI34.1
 of 9¯nitro-b-methylene-5-oxytetracycline in 200 ml of methanol and 1 ml of concentrated hydrochloric acid. The suspension is hydrogenated at atmospheric conditions until 3 moles of hydrogen have been consumed. Removal of the catalyst by filtration and evaporation of the filtrate gives a residue which is 9-amino-6-methylene-5-oxytetracycline.



  EXAMPLE XXXVU: .-
500 mg of 5% palladium on carbon are added to solutions.
 EMI34.2
 separate tions of 500 mg of lla-chlcro-6-deoxy-6-demethyl-6-methylenetetracycline hydrochloride and the corresponding lla-fluoro- derivative in 25 ml of methanol. Each suspension is then hydrogenated at atmospheric pressure until one mole of hydrogen has been absorbed. The catalyst is removed by filtration and the filtrate evaporated to dryness in vacuo. The separate compounds smell
 EMI34.3
 respectively 6-deoxy-6-demethyl-6-methylenetetracycline and a mixture of 11a-fluoro-6-deoxytetracycline and 11a-fluoro-6-6pi-6-deoxytetracycline.



  EXAMPLE XXXVIII. - 50 g of a mixture of the products lla-fluoro-6-deoxytetra-
 EMI34.4
 cyclin and lla-fluoro-6-epi-6-deooxytetracycline of Example XXXVII dissolved in 1.25 ml of 1% aqueous HCl, 30 g of zinc are added

 <Desc / Clms Page number 35>

 powder and the mixture is stirred for 15 minutes at 20 C. The zinc is removed by filtration and the product is extracted from the reaction mixture with butanol. After eva; oration to dryness, a mixture of 6-deoxytetracycline and 6-epi-6-deoxytetracycline is obtained.



  EXAMPLE XXXIX.-
 EMI35.1
 Salts obtained by addition of 6-e i-6-deao -c acid. tetrac cline 6-épi-6-désoay-5-oaytetracycline is dissolved in methanol containing an equimolar amount of hydrochloric acid.



  The hydrochloride is then precipitated by adding ether and is collected by filtration and dried. The hydrochloride is recrystallized from butanol.



   In the same way, other salts obtained by addition
 EMI35.2
 6-epi-6-deoxY-5-OxYtetracycline acid are prepared using, instead of hydrochloric acid, sulfuric, nitric, perchloric acid, hydrobromic, phosphoric, hydrofluoric, p-toluenesulfonic, hydroiodic, tartaric acids, acetic, citric, malic, benzoic glycollic, gluconic, gulonic, succinic and sulfosalycilic.



   The salts obtained by adding acid to the products mentioned in the examples below are also prepared in the same way. These salts obtained by addition of acid, which are not acceptable from the pharmaceutical point of view, can be used in the usual way for the purification in the preparation, either of amphoteric compounds or of their salts obtained by addition of acid.
 EMI35.3
 pharmacologically acceptable. In addition, salts obtained by addition of pharmaceutically unacceptable acids are useful as intermediates in the preparation of other products of the present invention as will be seen in the examples below.



    EXAMPLE XL, - Metal salts of 6-epi-6-deoxy-5-oxytetracycline
 EMI35.4
 The sodium salt of 6-epi-6-deoxy-5-oxytetracycline is prepared by dissolving the amphoteric compound in methanol containing

 <Desc / Clms Page number 36>

 taking an equimolecular amount of sodium hydroxide and evaporating the resulting mixture in vacuo. Other metal salts are prepared in the same way, including potassium, calcium, barium, lithium and strontium salts.



   The complex-metal salts of 6-epi-6-deoxy-6-oxy-tetracycline are prepared by dissolving it in a lower aliphatic alcohol, preferably methanol and treating it with an equimolecular amount of the chosen metal salt, dissolved. preferably in the chosen alcohol. The insoluble metal salt complexes are isolated by simple filtration. However, generally speaking, most of them are soluble in alcohol and they are recovered by evaporating the solvent or by adding a non-solvent such as diethyl ether.



   In this way, the metal salt complex of 6-epi-6-deoxy-5-oxytetracycline represented mainly by compounds containing the metal and 6-epi-6-deoxytetracycline in the ratio 1/1, are prepared by employing the following metal salts: calcium chloride, cobalt chloride, magnesium sulfate, magnesium chloride, stannous chloride, zinc chloride, cadmium chloride, barium chloride, silver nitrate, stannous nitrate, strontium nitrate, magnesium acetate, palladium chloride, magnesium chloride, cerium chloride, titanium chloride, platinum chloride, vanadium chloride, lead acetate, stannous bromide zinc sulfate, chromium chloride, and nickel chloride.



   And in the same way also the complex salts of the same metals of the products of the examples below are prepared.
 EMI36.1
 



  EXI14PLg¯UI 2 - 6-é i-6-déso -Ila-ehloro- -o tetrac eline 0.35 g of N-chlorosuccinimide is added to a cold and stirred solution of 1 g of 6-epi-6-deoxy- 5-oxytetracycline in 15 ml of dimethoxyethane. After 20 minutes at 0-5 C, the product is precipitated by adding ether, filtered and washed with ether.

 <Desc / Clms Page number 37>

 
 EMI37.1
 



  EXAMPLE XLII.-. 6-epi-6-deoxy-13a-fluoro-5-oxytetrac''ccline
A suspension of 4.8 g of 6-epi-6-deoxy-5-oxytetracycline hydrochloride in 100 ml of methanol is cooled to 0 ° C. while a stream of nitrogen is passed through it. 20 ml of a normal solution of sodium methoxide in methanol are added thereto and the mixture is saturated with perchloryl fluoride. The mixture is placed in the refrigerator for 24 hours, after which time the chlorate crystals of the desired product have formed. The filtrate still gives a second jet of less pure crystals.



  EXAMPLE XLV.-
 EMI37.2
 6¯éui-b-deox9 -? - choro-5-oxsrtétacvcline (1) 1 g of the product of Example XLIII is added to 50 ml of methanol containing 100 mg of rhodium on 5% carbon. The mixture is subjected to a pressure of 3 atmospheres of hydrogen at a temperature of 32 C. The hydrogenation is stopped when no more hydrogen is absorbed; the mixture is filtered and the resulting solution is evaporated to dryness under reduced pressure. The residue is triturated in 25 ml of ether, filtered and dried, to obtain the desired product.



  (2) Equivalent amounts of 6-epi-6-deoxy-7, 11a-dichlo-
 EMI37.3
 ro-5-oxytetracycline, sodium iodide and zinc powder are refluxed in acetone for 30 minutes. The reaction mixture is cooled and concentrated under reduced pressure to obtain the desired product.



  (3) 0.45 g of sodium hydrosulfite is added to a solution
 EMI37.4
 of 0.45 g of 6-epi-6-esoxy-7, 11a-dichloro-5-oxyte-tracycline perchlorate and the resulting mixture is stirred for 12 minutes. The product separates and is collected by filtration.



  EXAMPLE XLVI.-
 EMI37.5
 6-e i-b-deso -7- hén lazo-lla-chloro- -o tetrac cline
A molar amount of aniline is dissolved in AN hydrochloric acid, at a rate of 20 ml per gram of aniline, and this composition is treated with an equivalent of sodium nitrile at temperature.

 <Desc / Clms Page number 38>

 temperature of 0 C. The resulting solution of benzene diazonium chloride is mixed at a temperature ranging from 0 C to 20 C, with an aqueous solution of an equivalent of 6-epi-6¯deoxy-llachloro-5-oxytetracycline and d 'an equivalent amount of sodium hydroxide, this solution containing sufficient sodium carbonate to neutralize the excess hydrochloric acid present in the diazotized aniline solution. The pH of the resulting solution is of the order of 8 to 10.

   Stirring is continued at 0 C for about 2 hours, during which the 6-epi-6-deoxy-7-phenylazolla-chloro-5-oxytetracycline separates. This product is filtered, washed and dried.



  EXAMPLE XLVII.-
 EMI38.1
 6-epi-6-deoxy-7-amino-5-oxytetracycline
One part by weight of 6-epi-6-deoxy-7-phenylazo-lla-chloro-5-oxytetracycline is mixed with 20 parts by weight of methanol and 1/5 of a part by weight of 5% palladium on carbon (catalyst of hydrogenation) is added thereto. The mixture is then hydrogenated at 30-45 p.s.i. in a current device at 30 C until the hydrogen absorption ceases. The catalyst is then filtered off and the filtrate evaporated to dryness. The residue is a mixture of aniline and
 EMI38.2
 of 6-epi-6.lesaxy -? - a.a: .nü-5-oxytetracycline. The aniline is removed from the residue by washing it with ether.

   EXAMPLE XLVIII.-
 EMI38.3
 6-6pi-6-deoXV-7-.nl¯ro-5-oxytetracygline 4.58 g of 6-opi-6-deoxy-7-amino-5-oxytetracycline are dissolved in 20 ml of 6N nitric acid, at 0 C, and treated with 0.7 g of sodium nitrite. To this solution is then added a suspension of copper oxide in water, prepared by reducing an aqueous solution of copper sulfate (approximately 5 g in 50 ml of water) with an alkaline solution of glucose followed by 'neutralization with acetic acid. A reaction takes place in which nitrogen is released. The mixture is strongly acidified to decompose
 EMI38.4
 pose the complexes of copper and 6-epi-6-deoxy-7-nitro-5-oxy-

 <Desc / Clms Page number 39>

 tetracycline is extracted from solution with methyl isobutyl ketone.



   This compound can also be prepared at the same time as
 EMI39.1
 6-bpi-6-deoxy-9-nitro-5-oxyteracycline by direct nitration of 6-epi-6-deoxy-5-owtetracine. In practice, 0.96 g of ¯6-epi-6-deoxy-5-ox hydrochloride is dissolved; tetracyline at 0-5 C in 10 ml of concentrated sulfuric acid then treated dropwise with 1.4 ml of a 10: 1 solution of concentrated sulfuric acid and 70% nitric acid, stirring and maintaining the same temperature for a period of 15 minutes. The solution is then ver-
 EMI39.2
 dried in 400 ml of anhydrous ether and 6-epi-6-deoxy-5-oxytetracycli. Nitrated compounds precipitate in the form of their acid sulfate. The precipitate is filtered off, washed and dried.

   This crude nitration product is recrystallized. read in about 5 ml of methanol and give a mixture of 7-nitro-
 EMI39.3
 and 9-nitro-6-epi-6-d6soXY-5-oxytetracycline. The mixture is separated by partition chromatography on a column with the t, 'luene-pyridine-water system. Since the 7-nitro compound is the least polar, it is eluted first.



   The diazonium technique has the advantage of giving the 7-nitro derivative alone and of avoiding this separation.



  EXAMPLE XLIX. -
 EMI39.4
 6i-6-deoxy -? - acetv2amino-5-ōxtetracvline, 20 ml of acetic anhydride ¯¯ is added to a solution of 4.58 g of 6-epi-6-deoxy-7-arano-5-oxytetracycline in 200 ml of tetrahydrofuran and 200 ml of dry methanol, and the mixture is stirred for 45 minutes at room temperature. The mixture is then freed from most of the solvent and poured dropwise into dry ether, filtered and the solid cake is re-titrated in ether, filtered and dried to obtain the crude product. The crude product is chromatographed on a cellulose column and the purified product is obtained in the main fraction.



   Using the same process, the following products were prepared:

 <Desc / Clms Page number 40>

 
 EMI40.1
 0 ... 01; ' 1.-t> - t $ O ") ('."' "1 ':)!" ,, \ y 1 & .111 n ..'- II; ... t!:. ty tÁ t.rac) 'cl1n- Ó..Áp! -od,: s.) r: y .. "1.but'1 ry ,,' l'1o - I) t ".cylln 6 .. 6; 11. ..d S.:J r: y-? -prop1'ny la ltlno-li..ot8 tl "aCTclln. b;: i-t5-â, 'soxy-'wt'Lé3aatixo.-oxyrttraayclW e KOEùa, X1 , .b., - ..



  ¯ .. ", 3.., I" ".." '. Ic3' 'TS'al Add * 100 18 & of .'fotass1 \ .1l nitrate. \ << n "\ .;': ': u'H. of 1? G 1ft of 6-3pi-6-d6soXr-o3-oxytetracyclin * dana 5.0 ml decides flurrir1 ('Hle liquida anhyttre. Le;, ± mixture is Aclt4. 15.C making '40 1 ! t1nl1t .. t '-nIP11 t 8vapor4 dry 1 ...: -Lidu is triturated with ml of ether and nl.tré to obtain the crude product which is triturated in ...' aclde hydrochloride & 011 .. \ 1X to 5% and then extracted with OO1; 8no1. The butlhloic extract is concentrated to obtain the product (which is tecPiD% JliS4 in water and hydrochloric acid.



  Lm1 LI. = -Ci-b- é, c.: R> -. R -Qçi'ttracvci3n We have ,; or while stirring, at 20oC, .1, 3 g of powdered zinc at an uH-'m of 2.5 5 g of 6 - <! pi-6-deaoxy-9-nitro-5'-oxyt4tracyclitie in 50 1 al and 2 al of HCl c-'ncentI 4. After 15 minutes, the mixture is filtered, the fold is adjusted to 2.5 pt the mixture is extracted with butanol. The butanhydrate extract will concentrate to give the product i4stPé in the form of the hydrochloride.



  FPLF LI 6-ti-6-âs, z - 'c- c e
 EMI40.2
 20 ml of acetylformic diacid are added at room temperature.
 EMI40.3
 rature of the ice bath, to a solution of z0 g of 6-epi-6¯deoxy- '1ino-5-oxytetracycline in 40 mi of Pl1., A dry. The mixture is 1 at this temperature for 15 minutes then poured drop By drop? 1 liter of dry ether. The product is filtered, re-triturated in re-filtered ether. The crude product is suspended in 1 liter of water making a solution of pH 4.2 and stirred for 10 minutes for 0-foryrl aroupaents. The> H of the mixture is adjusted Rlr 55 5 t solution is, frozen and dried

 <Desc / Clms Page number 41>

 
 EMI41.1
 to give: <* crude product which is chromatographed on a cellulose column and the purified product is obtained in 1 <: fraction
 EMI41.2
 main.
 EMI41.3
 



  1),; 1ItICO, the following products were prepared: 6-6pi-6-deoxy-9-acéùyInmo-5-oxyté% raoyclin * -pi-t-àsoxr..9-butrryiamiao-5 oxtrracTaiae 6-n; -6-désary - proay2, antno5-axyttracycline 6-'pi-6-désaxy-9-vaiôry1-o-5-aqù qam * EXEMPL! MH.- 6-ui-6oaoxv - <) - chioro-5-ûTVt <tMColin <To a solution, cooled in an Ilace bath, of 1.04. C of corhyrate da b-âpi-b-déso-9-ato-j-osftilqtclaae in 5 al of water and 2 ml of aclete ehtorldriqte ocra08Vi. we add. , below the surface of the l1qu1de. 150 as 4th ni% P1% O of 80àiUQ in 2 mil of water, 1st having acted% 4, ... & an '20 ir "les \ Ml adds 50 mg of urn to destroy the exo% s 4" 014 . ni% ronX. Cuprous chloride / HCl is added and the mixture is kept at room temperature for 5 minutes. It was placed in a water bath at 0 ° C for 20 minutes, cooled, and extracted ... 0 from normal butanol.



  The extract is dried over 8Od1- and drssspoi-sed sulphate.



  J..8 cuprous chloride is introduced by adding a hot solution of 600 mg of copper sulphate pentab7dra ".. t 160 1111 of sodium chloride in 2 ml of water at 160 .. of 1d.n.lf1te of sodium in 1.7 ml of 1.lN sodium hydroxide. The mixture is cooled and the solid is washed with 1 ml of water, then dissolved in 1 ml.
 EMI41.4
 of concentrated HCl used above.
 EMI41.5
 Crystalline p-toluenesulfonate is prepared by
 EMI41.6
 usual technique.
 EMI41.7
 



  E.1CEMP lJE LIV.- 7 = ChIOrO-9-ni% ro-6-éPi-4ôsoxY-5-oxvtétrjczcime 220 g of gN03 are added to a solution of 1g of base 7-chloro-6-épi-6- 5-deoxy-oxytetracycline in 8 ml of anhydrous liquid hydrofluoric acid at 0 C. The mixture is stirred for 30 minutes

 <Desc / Clms Page number 42>

 at 0 C and the hydrofluoric acid is driven off with a stream of nitrogen.



  The residue is triturated in dry ether, filtered and dried to obtain the desired product.



  EXAMPLE LV.-
 EMI42.1
 9-ni v- ad co -.6- 1-6-deo -o tetracline 100 mg of KNO3 are added to a solution of 600 mg of 7ella-dichloro-6-6pi-6-deoxy-5- perchlorate ozytetracycline in 1.5 ml of anhydrous liquid hydrofluoric acid. The mixture is stirred at 15 C for 30 minutes and then evaporated to dryness. The residue is triturated in ether and filtered to obtain the crude product which is triturated in 5% aqueous HCl and extracted with butanol.



  The butanolic extract is concentrated and the product is obtained which is recrystallized from water and hydrochloric acid.



  EXAMPLE LVI. -
 EMI42.2
 9 -e -f-deso tetrac eline A solution of 5 g of lla-chloro-6-epi-6-deoxy-5-oxy-tetracycline in 10 ml of anhydrous hydrofluoric acid is left to stand for 3 1/2 hours , after which 1.2 g of KNO3 are added with stirring. Stirring is continued for a further 1 hour at the temperature of the ice bath and the mixture is poured into 200 ml of dry ether. The precipitate formed is filtered off, washed with
 EMI42.3
 ether for oben1 the crude product. LIJ'JDI'P1.B g L ..g, Kt.rcLb-ui-6-riéc, x 5-Q.xS'tétrac9cline Was added, while agibcnt at 20 C, 1.3 g of powdered zinc! The solution is 1.4 µg of 7-colioro-9-nitro-6-6pi-6-deoxy-5oIQ ”dissolved in 50 ml of water and 2 ml of concentrated HCl.



  After 15 minutes, the mixture is filtered, the pH adjusted to 2.5 and the ntlat "t '. 38 * 7t" it 5 times with: 30 l1Ù. of butanol by extraction.



  The combined butanol compounds give, after concentration, the desired 1iJ'e i in the form of the dihydrochloride.



  The same product is obtained by reductiü..1. with sodium hydreaulfite like 3-nl.tro-'7, lla-dichlc.o-6-épi-6d6svxy - oxydtracyc: l ine.

 <Desc / Clms Page number 43>

 



  EXAMPLE LVIII.-
 EMI43.1
 7,11a-dichloro-9-nit o-6-é i-6-deo -o tetrac cline 1 g of 7, ll-dichloro-6-6p-6-deoxy-5-oxytetracycline- hydrochloride is dissolved. in 20 ml of acetic acid and 1 ml of concentrated HNO3 is added. The mixture is allowed to stand for 12 hours and then concentrated to 1/4 of its original volume. It is added dropwise to 200 ml of ether with stirring at the temperature of the ice bath. Stirring is continued for 3 hours and the solid is then recovered by filtration. The solid is triturated several times in ether and then dried to obtain the product in the form of the hydrochloride.



   Using the same process, the following products were obtained from the corresponding starting compounds;
 EMI43.2
 7-ehloro-lla-fluoro-9-nitro-6-épi-6-deoxY-5-oxytetracycline 7-brano-lla-chloro-9-nitro-6-épi-6-desc: xy-5-oxytetracycline 7 = iodo-lla-fluoro-9-nitro-6-epi-6-deoxy-5-oa3'tetracycline These products are converted to 7-ehlro-, 7-bromo-, and 7-iodo-9-amino-6-, respectively. 6pi-6-deoxy-5-OxYtetracyclines by the reduction methods of the previous examples. EXAMPLE LIX.-
 EMI43.3
 fo 1 n - -c oro-6- i-6-deo -o tetracvcline A a solution of 4.5 g of the base 9-amino-7-chloro-6- epi-6-deoxy-5-oxytetracycline in 40 ml of dry pyridine, 20 ml of acetylformic acid are added at the temperature of the ice bath.

   The mixture is stirred for 15 min at this temperature, then added dropwise to 1 liter of dry ether. The product is filtered, triturated in ether and again triturated. The crude product is then suspended in 100 ml of water producing a solution of pH 4.2 and stirred for 10 minutes to hydrolyze the C-formyl groups. The pH of the mixture is then adjusted to 5.5 and the solution is frozen and dried to obtain a crude product which is chromatographed on a cellulose column to obtain the purified product in the main fraction.

 <Desc / Clms Page number 44>

 



  EXAMPLE LX. -
 EMI44.1
 -acetvlamino-7-chloro-bib-deoxy- -trr = --- cvçline To a solution of ±, 2 g of the base ï-c.hloro- -a: nir: - epi-6-deoxy-5 -oxytetracycline in 200 ml of dry detéti'xt; ydr7iuran-ne and 200 ml of dry methanol, 20 wl of anhydrl <1t- 'ac4tiq e are added and the mixture is stirred for 45 minutes at temperature a, nblante. The mixture is then freed from most of the solvent and poured into dry ether, filtered and the solid cake is re-titrated in ether, filtered and dried to obtain the crude product. The crude product is chromatographed as in Example LIX to obtain the product purified in the -rincipale fraction.
 EMI44.2
 



  Using the methods of Example LIX set from E'x '>' 1le present, the following products are obtained: 9-butyamino -? - chloro-6-ep.b-des, xy-5-oxyté tracy cl ine 9-propionylamino-7-chloro-6epi-6-deoxy-5-oxytetraglycline 9-valeryaminO: 7-chloro-6-epi-6-deoXY-5-oxytetracycline EXE1P LE LXI.'7-A suspension of 6- epi-6-deoxy-5-oxytetracycline is prepared with the following compositions antibiotic 31.42 g aqueous sorbitol 70% 714.29 g glycerin USP 185.35 gum acacia (10% solution) 100 ml polyvinyl pyrrolidone 0.5 g butyl parahydroxybenzoate (preservative) 0.172 g propyl parahydroxybenzoate (preservative) 0.094g distilled water, to obtain 1 liter To this suspension can be added to choice of different sweetening and flavoring agents, as well as acceptable colorings.



  The suspension contains approximately 25 mg of antibiotic activity per liter.



  EXAMPLE LXII, -
A suspension of 6-epi-6-deoxy-5-oxytetracycline is

 <Desc / Clms Page number 45>

 prepared with the following compositions: antibiotic 30.22 g magnesium chloride hexahydrate 12.36 g
 EMI45.1
 wonoethanolamine 8.85 ml propylene glycol 376 g ea 94 ml
 EMI45.2
 s 1 has a concentration of 50 mg /: nl and is suitable for acLnin -1. F - tion -ntArAle and sp4etaleinent for cintramuscular administration.



  E,> ::; <:; 1t 'J., E .weI H. - A compound base prepared by mixing the following ingredients: in pr @ criions indicated by weight: sucrose U.S.P. 80.3
 EMI45.3
 a donation of tapioca. 13, 2 magnesium stearate 6.5 On this basis, surf1sa! N.! N'nt of 6-Pi-h¯.PS, -, xyttracycii.ne is mixed to obtain tablets containing? 5, 100 and 25 < )) 7 active ingredients.



  EXAMPLE LXIV, -
A mixture is prepared containing the following ingredients: Calcium carbohydrate U.S.p. 17.6 g dicalcium phosphate 18.8
 EMI45.4
 trisilic> x: .e duo magnesium U.S.P. 5.2 U. @. @. Lactone 5.2
 EMI45.5
 POltU starch; -, O '? earth 5.2 magnesium stearate @ um A 0.8 magnesium stearate B 0.35
 EMI45.6
 Tan added to this mixture zfisa'7ment of 6-epi-6-dcsox, -h-oxftcira-cyclin to obtain towels containing 25, 100 and 250 mg of active ingredients. EXAMPLE LXV. -
 EMI45.7
 1000 g of 6-epi-6-deoxy-f: -oxytetracycline are thoroughly mixed and ground with 2500 g of 900i1111 tcoroate.

   The ground and dry mixture is poured into t'l.: I, as, st4rilLs ±> with

 <Desc / Clms Page number 46>

 
 EMI46.1
 ett1 'oxide:, lr..ne and the strands sent 5 (' ell. "'t> s' ho fa.on sterile.



  For i'ad! .... iatravettneuse istr3tion, or aJout = 3ufr1q'W1!) ". L? Nt water to flasks WH ..." obtain a solution containing 0 mg f: tnr = il.errt active by .He.
 EMI46.2
 RESELL r C A T I 0 ii S. ------------------------------ 1.- Process for the preparation of a 6-deoxytetracyclinc, characterized in that hydrogen is a compound having the formula I or II given above, in the presence of a noble metal catalyst.



   2.- Process for the production of a mixture of a 6-
 EMI46.3
 desorr-5-oxytetracycline and a 6-epi-6-deoxy-5-oxytetracyc; ine, characterized in that a compound having the formula I or II given: 'above is hydrogenated, where the substance Y is OH, in the presence of a noble metal catalyst.


    

Claims (1)

3. A method according to claim 1 or claim EMI46.4 ti m 2, characterized in that the catalyst of mptpl nobip is rhodium or pailadi'n. 4. A method according to any one of the preceding claims, characterized in that the hydrogenation is carried out at a pressure between one atom and 2000 pounds square inch (140 kg / cm2). 5. - Process according to any one of the claims EMI46.5 Preces, characterized in this bare the temperature used is included in .-). and 60 C. 6.¯ Pro according to any one of the preceding claims, aractérls that the compounds 6-deoxy- and 6-epi- 5..dbsnç, rttsacycline are obtained -iréraent. 7.- Process according to -> 'one that \ 3ccr .., of the preceding claims, characterized in that utip one .1oursuit -'- à6 -1 ognation until reaction of 1 to 6 moles of hydrogen per mole of coi, ., starting. EMI46.6 8.- Process the i production ries, .7s-h.lo-5pi .- <isooy-5¯ oxytétrl1cyc.Lines, character? Rlsr fm that surely in cnncirt uno <Desc / Clms Page number 47> EMI47.1 6-epi-6-deso, -5-oxyté.tracycline with a halogenating agent in an inert solvent. 9. - Method according to claim 8, characterized in EMI47.2 that the halogenating agent is chlorine; N-chloro-alpha lower alkyl acid acids; imides of '-nydrocaibonés-dicarbox-yli c acids or N lower alkyl anilides; ! --- 3-chloro- or 3.5cii; Antoine's 5, 5-dimethylhT; ridinium perchloride alkhydrates; lower aikyl hypochlorites; or perchloryl fluoride. 10. A method according to claim 8 or 9, characterized in that the halogenating agent is used in amounts ranging from 1 to 1.2 moles per mole of tetracycline. 11. A method according to either of claims to 10, characterized in re that the halogenation is carried out in the presence of a water-miscible solvent. 12. - Process according to claim 11, characterized EMI47.3 in that the halogenating agent eii di; perchloryl fluoride and the solvent 1, n alkyl alcohol infr: .r. 13.- Procr'dé 1 ree.-dication 12, characterized in that the iiuoru1 ation is carried out in; JréE'f1Cf> of a base. I4.- r'ror.clrs s, ivarit 1 '' -'Pt '' u 1-1, K. tre dPs claims 8 to 13, characterized in that '(ùf-' .'1; a; <g <'1. veined b't tion µ 1 = ne temperature below 5 (' oC. 15.- Prncéoé following ia r ('' I'f'J <lLca + 1,> n 8 0'1 .. '9, crira-.tér-àls, 7 in that the agent ha1ogpnrd1t is pn'rï0 : r. '"; 1:', 5 of rropt. Gropings of at least 2 moles per mole of tetracycline. 16.- Prcced according to 'ur; e, "pJ ccnl -., 1 - ,,? Of the previous revP: .c? .CaW.r: s .. characterized in that @@' further treats the product obtained by a nitration agent. EMI47.4 17.- A fa-cpi-c-fscxy.-5-Gtctrac: = clir.e substlt.u <'e on cycle E, in lrquc = 1.1E l 51 bstituent is a chlorine, a hyrir0Fi.e or a nit! 'n, 4ml,,' 111, Ütyl-t!: ', l1'1 (,) irùSrir1ar group. 18.- Un (lr1 ....; ii-chloro- '' ''. ', ILIi) 3'Cf-- tif-' j> r <> <1I n; '.P the rErt vpr; j, cÉa t i c) 11 17. <Desc / Clms Page number 48> EMI48.1 11.- 6-cpi-6-deoxy-5-o: xytetracycline. 20.- 6-epi-6-deoxy-7-chloro-5-oxytetracycline. 21.- 6-epi-6-deoxy-11a-fauoro-5-oytetracycline. 22.- 6-epi-6-deoxy-9-nitro-5-oxytetracycline. 23.- 6-epi-6-deoxy-9-s.rüino-5-oaytetracycline. 24.- 6-epi-6-deoxy-9-acetylamino-5-oxytetracycline. 25. - 6-epi-6-deoxy-7-chloro-11a-fluoro-5-oxytetracycline. 26.- 6-epi-6-deoxy-11a-chloro-5-oxytetracycline. 27.- The salts of the compounds of any one of claims 17 to 26. 28. - A product according to claim 17, in substance EMI48.2 as above. described. 29. - A product according to claim 18, substantially as above. described. 30. - A method according to claim 1, in substance as described above. 31. - A method according to claim 8, in substance as described above.