IL27223A - Tetracycline derivatives,a process for their preparation and therapeutical use thereof - Google Patents

Description

C O H E N Z E D E K S P I S B A C H R E G D PA TE NT A TT O R N EYS LEVONTIN ST 1169 1 E AV I V P A T E N T S D E S I G N S O R D I N A N C E SPECIFICATION Dgfi A PROCESS FOR PREPARATION ND USE Cia an Italian Joint Stock of Vie DO HEREBY DECLARE the nature of this invention and in what manner the same is to be performed to be particularly described and ascertained in and by the following This application relates to novel and useful tetracycline derivatives having particular and to a process for their preparation as well as to their in It is known that tetracyclines react with formaldehyde and primary and secondary amines 1959 German Patent 044 and with formaldehyde and carboxylic aminoacids armaceutico Belgiaa Patent 5 and with sole formaldehyde Patent 1 to give water soluble An object of the present invention consists of novel condensation products of with formaldehyde and a particular type of amines containing a sulfuxated acid such products being never before described and having revealed surprising pharmacological Another object of the invention consists of the preparation process of the products which are the primary object of the present A further object of the invention consists of the as components of pharmaceutical particularly as antibacterial The compounds of this invention are reaction products of a tetracycline with formaldehyde ana a particular type of amine having a sulfurated acid function in its molecule characterizea by the following general wherein can De hydrogen or oxyalkyl with no more than carbon Z is or X is OH when Z or X is ii or SH when Z deriving from this general formula and useful to the invention are for example thane sulfonic me e hane sulfonic ethylaminome thane sulfonic uhane sulfonic methane sulfonic thane sulfonic any those which can be obtained varying the general formula within the previously defined When the above amines are reacted in suitable conditions with formaldehyde and a there are obtained the compounds which are the of this As to the molecular constitution of said it can be stated that elemental analysis to this chiefly that of S is being the new characteristic indicates in these novel derivatives a molar ratio amine tetracycline of The infrared spectrum indicates the presence of the band of secondary which would demonstrate that tetracycline is bound to the amine through the group by means of a methylene since the molar ratios between amine and tetracycline are respected with both the amines where H as well as those where or hydr this suggests that a molecule of tetracycline is bound to the amine as and the second molecule of tetracycline has a different not yet definitely determined The above constitutes our latest view on the intimate molecular constitution of the products of the a constitution which had been previously p ovisionally briefly established on the basis of preliminary approximative In any with regard to the structural we do not intend to bind ourselves with theoretical the important thing being chiefly the fact that operating according to the process which is the object of this invention tnere are obtained products which have a definite and constant for which the infrared spectrum shows always the above mentioned absorption and the elemental analyses vary merely within the limits of the experimental errors and any way indicate a molar ratio tetracycline which is the above that is Tetracyclines particularly useful to the invention hlorotetra eye line also as well as their can be transformed according to the process of the In the reaction the acid group from the amine having a sul urated acid can be free or salified with potassium or an including antibiotics of a basic nature as kanamycin the products salified with sodium are The novel derivatives are generall very water soluble and some of them show a stability in consequently they lend themselves very well to be used by both mouth as well as parenteral since they are well absorbed no matter the way of adminis ration In the following tables there are indicated some properties of some preferred products of the it is to be understood that we have followed similar experiments also with the other products which are the object of this and we have obtained qualitatively even if quantitatively results Table 1 Protection by mouth of white rats infected with Klebsiella pneumoniae All expressed as of chlor hyd in order to allow a direct Product in m Chlo otetracycline hydrochloride oxyethyl The reaction product between methane sulfonic formaldehyde and chlo tetracycline according to this 58 The Reaction between thane sulfonic formaldehyde and cycline according to this bO The reaction product between 1 mi ethane thiosulfonic and tracycline according to this The acute and chronic toxicities are lower not only in comparison with the starting antibiotics example chlor otetracycline but also in comparison with already known derivatives of such antibiotics 2 Intravenous administration to Swiss white All are expressed as chlor ote racycline hydrochloride in order to allow a direct Product Chlo ote racycline hydrochloride otetracycline Patent 1 196 117 The reaction product between thane sulfonic formaldehyde and chlor cycline according to this The reaction product between thane sulfonic formaldehyde and tracycline according to this 142 The reaction product between thane thiosulfonic formaldehyde and chlor tracycline according to this 138 Table 3 Chronical toxicity Intraperitoneal adminis ration to Swiss white mg per body Doses are all expressed as otetracycline Mortality 10 days days Chlor otetracycline hydrochloride The reaction product between aminome thane sulfonic dehyde and chlor otetracycline according to this The reaction product between sulfinic formaldehyde and chlo otetracycline according to this The reaction product between thtosulfonic formaldehyde and tracycline according to this Table Chronical toxicity Administration by mouth to Swiss white mg per kg Doses are expressed as tetracycline hyd ochlo Average weight od uc the owing 50 Chlor otetracycline chloride 21 22 The reaction product between thane sulfonic formaldehyde and c or o tracyc line according to this 21 The reaction product between thane thiosulfonic formaldehyde and chlorotetracycline according to this Untreated control animals 24 To these evident and remarkable representing per se already a progress in the present state of there are added other ones which are at all surprising and absolutely novel for the antibiotics of the tetracycline It is known that among the chief inconveniences due to tetracyclines there is a liver toxicity due to a fatty degeneration of particularly with tetracycline and oxyte acyc and this toxicity may lead in certain cases to the 523 New England of 99 A of Internal Medicine 271 To no tetracycline derivative has to such a severe side on the contrary with the products of this invention we have obtained surprising with a considerable reduction of t fat percent in The following data are a demonstration of the above Table 5 Percent of fat in liver Dose of 100 per Kg intravenously to Wistar white The percentages represent the increase of fat in comparison with untreated control Doses are expressed as chlo otetracycline hyd ochloride Percent of fat in liver 48 and 2 hours after P oduct the drug administration 48 hours hours Chlo otetracycline hyd ochloride bO The reaction product between sulfinic and chlo otetracycline according to this The reaction product between thane sulfonic formaldehyde and chlorotetracycline according to this These novel products are prepared by a process which consists tically of the condensation reaction of the starting substances which have been aiscussed the that is a formaldehyde and aa amine containing a sulfurated acid The condensation can be effected in various in solvent types which do not take part ta the such as isopropyl lf tetrahydrof both per se as well as in the mixture one The molar ratios of the reacting substances can be varied within fairly wide however the preferred reaction ratios amine ormaldehyde tetracycline range from to In such a for every the optimum of the reaction may be o can be employed indif in aqueous in methanol solution or as a however the preferred form is thejaqueous wnich is also the cheapest Tetracycline can be utilized both as an amphoteric as well as under the form of an addition salt with an acid the hydrochloride or under the form of an alkaline salt sodium The reaction temperature can range from to an optimum temperature is comprised and According to the product type and the reaction the product can separate spontaneously during the course of the or it can be isolated by it with wherein it is not or by concentrating the reaction mixture by means of the solvent distilla The following illustrative serve to make clear the method and to demonstrate how one can vary the conditions within the previously stated it is clear to the experts in the art tnat these examples can be simply extended also to not described but falling in the scope of this invention by small routine The do not coustitute any limitation to the Example 1 g ha OH and aminome hane sulfonic acid are dissolved in of a mixture of methanol and aqueous 0 formaldehyde after 15 tetracycline dissolved in 80 are After some a precipitation stirring is continued still for minutes at the product is washed with methyl alcohol and dried at under reduced It is a light yellow very soluble in an aqueous solution has a pH of Analysis for over under high calculated C N Found C 5 H S Example 2 aminome tha e sulfonic acid is dissolved in 80 methanol containing there is 10 chlorotetracycline dissolved in methanol and dimet o The mixture is filtered in order to avoid possible insoluble and there is added aqueous and the mixture is left then to react for 60 mxnutes at room After some minutes the product it is washed with methanol and dried at under Analysis for calculated S found 3 3 thane sulfonic acid aOfi are dissolved in 60 ml to the solution there is added aqueous f the mixture is left to react for 5 then there is added tetracycline dissolved in 60 ml and 40 ml the mixture is kept under stirring at room temperature for minutes and during this time tne separation of the product takes which is collected and washed with opr The analysis for has calculated Example and sulfonic acid are dissolved in 110 after adding aqueous formaldehyde the mixture is unaer stirring for 10 and then there is 10 g aissolved in a mixture of 60 ml ml after some minutes the product precipitates It is left still for minutes to then it is filtered and washed with For calculated S found Example After having dissolved and g e hylaminome sulfonic acid in 40 to the solution there is adaed successively aqueous after cycline aissolved in a mixture consisting of isopropyl alcohol methanul and ormamide The whole is left to react at room temperature for minutes and then the separate product is For calculated found and Example 6 aqueous yde is aaded to a solution of mole sodium salt of thane sulfonic acid in ml after 5 minutes there is added a solution of cnlo otetracycline in a mixture of 100 ml isopropanol and d ime thy mid e The whole is left under at room temperature for 30 minutes and then the separated product is filtered washed with isopropyl alcohol and dried at unaer 7 and oxyethylamino ethane sulfonic acid are dissolved in 120 ml there is added ml aqueous f or Id e the mixture is left for 15 minutes at room temperature and chlor dissolved in 60 ml d i me thy If or ma mid e and ml is added to the and allowed to react for minutes at The reaction precipitates is then washed with isopropanol and dried under vacuum at The product is very water an aqueous o solution has of An analytical dried over under high vacuum has given the following for a found CI calculated CI Example 8 g g thane sulfinic acid and 3 aqueous 40 formaldehyde are dissolved in 160 ml methanol at room mixture is left for minutes at room temperature and then there added tetracycline dissolved in 80 ml It is stirred minutes at and then the precipitated product is filtered washed with methanol and dried under vacuum at 4 The product is water a aqueous solution has pH of In it shows absorption peaks at 0 and The rotatory power is An analytical dried under high vacuum over during some yields the following values for calculated C H S found C H S Na The antibacterial activity in vitro against Sarcina while using tetracycline hydrochloride as the is of Example 9 and sulfinic acid are dissolved in the solution is treated with aqueous f ormaldeh de and the mixture is left for 5 at room is added g chlorotetracycline the dissolved in 80 ml ormamide and 60 and the mixture is left to react for minutes at the product separates spontaneously and is washed on the filter with methanol and dried under vacuum at It is very water A solution has a pH between and In N NaOH it shows absorption peaks at 253 2 The rotatory power is in The infrared spectrum presents the 1526 band of ituted For the elemental the sample was dried under high vacuum over For Calculated G CI S Na C and H and N and CI and S Na and The antibacterial activity in vitro against while using chlorote racycline cnloride as the is of g example 10 g and g tha e thiosulfonic acid dissolved in 80 ml are treated with aqueous 4 after 5 with a solution of 9 g tetracycline he dissolved in 40 ml while leaving under stirring at room temperature a yellow product which is filtered out after washed with methanol and dried under A na lys is f or found C H Na calculated C S An aqueous solution has a pH In N it shows absorption peaks at 270 and 382 The in e antibacterial activity in vitro against while using tetracycline hgrdr ochloride as the is of Example 11 mole sodium thane thiosulf onate is reacted for 5 minutes at room temperature with aqueous formaldehyde in 80 ml there is added a solution of 10 tetracycline in 40 ormamide and ml After some minutes the product the reaction mass is held for a total of minutes at and then it is the product is washed with methanol and dried under Analysis for found C H CI S calculated C H N CI S Ha An aqueous solution has a pH a a pH In N it shows absorption D peaks at 253 285 rotatory power is 0 1 in The infrared spectrum presents the of monosubstituted The antibacterial activity in vitro against Sarcina while using chlo tracycline the is of 900 12 g ox and hane sulfinic acid are dissolved in a mixture of 60 ml methanol and ml methoxyethanol after all substances are the mixture is filtered and ml aqueous formaldehyde is added to the after some minutes the reaction product The mixture is stirred for JO minutes at room then the product is washed with methanol and dried at under reduced For C calculated found S 13 To a solution of mole sodium ethane sulfinate in ml methanol there are added ml aqueous formaldehyde after five a solution of cline prepared by suspending 5 demeth lchlo o e racycline hydrochloride in 20 ml and neutralizing it with g in 25 ml The mixture is left to react at for minutes and the separated product is filtered It is dried under vacuum at Analysis f or calculated S Example 14 NaOH and thane thiosulf acid are dissolved in 40 oil there is added oxyte chloride in ml after the substance is the mixture is filtered to eliminate possible present then the filtrate is diluted with ml isopropanol and treated with ml aqueous It is stirred for 60 minutes at room After some minutes the product begins to it is washed with isopropanol and dried under a reduced pressure at found S gxample 15 A solution of mole sodium thane thiosulf ona e in 40 ml methanol is reacted for 5 minutes at room temperature with aqueous formaldehyde and then is treated with a solution of mole bhyltetracycline dissolved in 20 ml oy and 25 ml after some minutes the reaction product The reaction mass is stirred at room temperature for a total of then it is filtered and the product is washed with After drying over under the product yields the following data on analysing for calculated found A solution of sulfinic acid and aqueous formaldehyde in 5 methanol is To this solution there is added a second one prepared from g tetracycline hyd ochloride suspended in 20 ml ha ol and neutralized with g NaOH in 15 ml The resulting solution is diluted with 20 ml isopropyl alcohol and left to react at room temperature for JO the separated proauGt is filtered washed with methanol and dried under vacuum at found calculated Example 17 g sulfinic acid and ml aqueous formaldehyde are dissolved in ml a separated suspension of g cycline hydrochloride in a mixture of ml methanol and 40 ml is neutralized with g NaOH dissolved in methanol and the resulting solution is filtered and is added to the solution containing the and then it is le to react for minutes at The precipitated product is washed with isopropyl alcohol and dried at under a reduced a 1 The product is identic to that of iixample Various analyzed samples 18 To a solution containing aqueous and mole sodium sulfonate in 60 ml methanol there is added a solution prepared by neutralizing S cycline hyd ochloride suspended in 40 ml dimethy with in ml it is left under stirring for 30 the precipitated product is filtered and dried at under calculated The product is equal to that of Example Example 19 hanolaminome thane sulfonic acid and ml aqueous formaldehyde are dissolved in 160 ml After 5 minutes there is added 20 chlorotetra c cline dissolved in 80 ml and ml the resulting solution is held for 1 hour at filtered to eliminate possible undissolved and diluted with ml isopropyl the precipitated product is washed on the filter with isopropyl and dried under vacuum at Various analysed samples gave the following results calculated S found S insufficientOCRQuality

Claims (1)

1. NOW particularly described and tained the nature of our said invention and in what manner the same is to be we declare that wha we is A process for the preparation of novel derivatives of antibiotics of the tetracycline comprising the reaction in a suitable solvent of a tetracycline with formaldehyde and an amine containing a sulfurated acid function having the following general wherein B can be hydrogen or or hydroxyalkyl with no more than carbon Z is or X is when Z or X is H or when Z is A process according to Claim comprising the reaction of 1 to moles formaldehyde and 1 to 2 moles tetracycline per every mole A process according to Claims 1 and comprising the reaction of the amine containing a sulfurated acid as defined in Claim under the form of a sodium A process according to Claims 2 and comprising the use of methanol in mixture with t o mid or as the A process according to Claims 2 and comprising the use of a mixture o amide or opr as the A process according to Claims 1 to comprising the use of formaldehyde in an aqueous s A process according to Claims 1 to comprising carrying out of the reaction at a temperature between and A chemical compound characterized by the fact that it is obtained by a process according to Claims 1 to when a tetracycline selected from the group consisting of and e racycline is reacted with formaldehyde and an amine containing a sulfurated acid function as defined in Claim A compound according to Claim wherein the antibiotic is tetracycline and the amine is sodium a ane s A compound according to Claim wherein the antibiotic is chlor otetracycline and the amine is sodium a t hane s one t e A compound according to Claim wherein the antibiotic is chlor otetracycline and the amine is sodium A compound according to Claim wherein the antibiotic is chlo otetracycline and the amine is sodium e hane A compound according to Claim wherein the antibiotic is tetracycline and the amine is sodium laminome thane 3 A compound according to Claim wherein the antibiotic is chlorotetracycline and the amine is sodium me th e compound according to Claim wherein the antibiotic is chlorotetracycline and the amine is sodium A compound according to Claim wherein the antibiotic is tetracycline and the amine is sodium sulfinate A compound according to Claim 8 wherein the antibiotic is chlo otetracycline and the amine is sodium ethane inate A compound according to Claim 8 wherein the antibiotic is tetracycline and the amine is sodium onate A compound according to Claim wherein the antibiotic is and the amine is sodium thiosulf ona te A compound according to Claim wherein the antibiotic is ox tetracycline and the amine is sodium thane A compound accordin to Claim wherein the antibiotic is and the amine is sodium A compound according to Claim wherein the antibiotic is eye line and the amine is sodium tha e ona A compound according to Claim wherein the antibiotic is and the amine is sodium ona te A drug composition characterized by the fact that it alone or in association with other one or more compounds according to Claims 8 to Attorneys or Applicants insufficientOCRQuality