PH26472A - A method for the prophylaxis or treatment of infections using benzylamine derivatives - Google Patents

Description

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ANTI-ADHESIVE FROPHYLACTIC AHD FHARMACEUTICAL

COMPOSITIONS CONTAINING BENZYLAMINE DERIVATIVES

U.S. latents Nos. 3,336,308, 3,536,713, and 4,113,777 describe benzylamine derivatives ~ and the pharmacologically acceptable acid addition salts thereof with inorganic or organic acids which have secretolytic properties, in particular, in addition to an anti-tussive etfiect. The two compounds bruihexine hydrochloride [N-methyl-N-(2- amino-3,5-ditromotenzyl)-cyclohexylamine hydro- chloride/ nd ambroxol hydrochloride [/F-(?-amino- 3,5-dibromobenzyl )-trans-4-hydroxy-cyclohexyl- amine hydrochloride/ have been used as secretoly- tics lor therapeutic purposes, 1t has now been surprisingly found that benzylamines of the tormula

Me

Br CH, - I :

NC (1) i". . it > 1 ir wherein

Ry represents a nydroxyl group in the 2- or 4-pcsitlon or an amino group in the 2-poslition; : - 2 -

BAD ORIGINAL

— anna fat. 2647% 26472

Ry, Tepresents a hydroRen atom or a methyl group; and

Rs represents a cyclohexyl or hydroxy- cyclohexyl group, and the pharmacologically accept- . 5 able acid addition salts thereof with d¢norganic or organic acids, inhibit or prevent the colonigation of surfaces by microorganisus, particularly in the oral and pharyngeal cavity, in the lungs, in the urogenital tract, in the lnammary glands, on the oo skin, and in the conjunctival membranes,

Preferred compounds of Formula 1 are those wherein R, and Ry together with the nitrogen atom between them represent the N-methyl-cyclohexyl- amino, cis-3phydroxy-cyclohexylamino, or trang-4¢ hydroxy-cyclohexylamino group. ihe compounds

N-methyl-N-(2-amino-3,5-dibromo-tenzyl )-

Ccyclohexylamine,

N-(2-amino-3, 5-dibromo-benzyl)-trans-4- hydroxy-cyclohexylamine,

N-(3,5-dibromo-4-hydroxy-benzyl )-cig-3- hydroxy-cyclohexylamine, and

N-(3,5-d1ibromo-2-hydroxy-benzyl)-trans-4- hydroxy-cyclohexylamine, : and the pharmacologically acceptable acid addition salts thereoil with inorganic or organic acids, are especially preierred.

. ful. 2472

The compounds of Formula I may further- more be converted into their addition salts, especially into thetr pharmacologically acceptable salts with inorganic or organic acids. Suitable acids may be, for example, hydrochloric acid, hydrobromic acid, gulfuric acid, phosphoric acid, lactic acid, citric acid, tartaric acid, succinic acld, maleic acid, or fumaric acid.

Thus, according to the invention, by applying a compound of Formula 1, the risk of in- : fection uy microbes is reduced or eliminated by a reduction in the ability of the microbes to adhere to the cell walls. This is particularly true with regard to the oral and pharyngeal cavities, the lungs, vagina, and mammary glands. ror example, this prophylactic activity of the compounds

A = N-methyl-N-(2-amino-3, 5-dibromo- bengyl)-cyclohexylamine hydrochloride and

B = N-(2-amino-3,5-dibromo-benzyl)-trans- 4-hydroxy-cyclohexylamine hydrochloride was tested as follows: 1. Direct activity in the in vitro system (a) The titre of the living cells, e.g.» human epithelial cells from the lungs such as ATCC, .

fat. 26472

CCL5, L-132 (with which the compounds of Formula I have been shown to have a very close affinity com- parable with that of the intact organ) is deter- mined by vital staining with trypan blue and count- ing in a Fuchs-Rosenthal Counting chamber and by : diluting with culture medium, e.g., iFinimum Essential

Medium plus 10% foetal calves' serum plus 0.1% kanamycin, adjusted to 7 x 10 cells/ml. Three milliliter portions of tiis suspension are poured into glass scintillation vials which have been prepared by the usual treatment for the adhesion of epithelial cells to the surface of the glass. : they are incubated for three days at 37°C with 3%

Co, until a sealed monolayer is formed. A monolayer corresponds to 5 x 10° cells per scintillation vial. (b) The bacteria, e.g., Klebsiella pneumo- niae Weingarten 14, are cultured in Brain Heart

Infusion Broth (BHI) in two passages lasting 24 hours each at 37°C until the logarithmic growth phase is reached. During the second passage, radio- active labelling is carried out by the addition of 1,2-*4c-acetate to the culture medium. Then the culture is washed twice with PBS (phosphate bufferred saiine) to eliminate any non-incorporated radioactivity. Using PBS of pH 5,5, the culture is adjusted to a transmission of 35 % in a photometer, corresponding to a titre of 5 x 108 bacteria/ml,

ful. 26¢72— by determination of the number of living pathogens. (¢c) The bacterial culture is centrifuged, and the PBS ie taken off and replaced by the same volume of benzylamine solution in PBS of pH 5.5.

For the control, FBS of pH 5.5 is used instead of the benzylamine solution.

The culture medium of the cells is removed, the monolayer (= 5 x 10° cells) is rinsed twice with PB5 of pi 5.5%, and is covered with 1 ml of bacterial benzyiamine solution (z= 5 x 108 baceteria).

Incubation is carried out .or three hours at 37°C,

Then the bacterial benzylamine solution is removed, and the monolayer 1s rinsed twice with

PBS of pli 5.5 to remove any non-adhering bacteria.

After addition of INSTAGrL (registered trade mark of Messrs. Fackard, USA, the composition was not disclosed) the radioactivity of the bonded bacteria is measured in a scintillation counter. (d) Hesults: alth Compound A (50 pg/ml) 1Y % fewer bacteria were detected on the epithelial cells, by comparison with the control, while with Compound

B (50 ug/ml) 20 % fewer were found. 2, Indirect activit in the ex vivo system (a) The epithelial cells are grown as described above.

fof. 76473 (b) The bacteria are grown as described above, (¢) Male SPF rats (Chbb:THOM) are treated orally for five days with 25 mg of benzylamine per kg of body weight or with TYLOSE (R) a water- soluble cellulose ether available trom the

Hoechst Corp., as control. Lung lavage material is obtained vy brouchoalveolar lavage with isotonic urea solution and then pooled. Une hundred milli- liters of thie iluid are divided into 30 protein- containing gel fractions by preparative isoelectric focussing. After the isdelectric point of indi- vidual fractions has been determined with a surface electrode, the proteins are eluted from the gel with 3 ml of PBS, pil 5.5, for each fraction. The 30 fractions are adjusted to a pH of 5.5, (d) The bacterial culture is centrifuged and PBS is removed and replaced by the same volume : of bronchoalveclar lavage fraction. For the control, instead of the bronchoalveolar lavage fractions,

PBS of pH 5.5 and isotonic urea solution, treated in the same way, are used. After two hours, incubation at 57°C the bacterial culture is centri- fuged again, bronchoalveolar lavage fractions or

PBS or urea are removed and replaced by the same volume of PBS of pH 5.5.

od poy7-

The culture medium of the cells is renoved, the monolayer (= 9 Xx 10° cells) is washed twice with PBS of pH 5.5, and 1 ml of bacterial sus- pension (= 5 x 10° bacteria) is poured dver it.

Incubation is carried out tor two hours at 37°C.

The bacterial suspension 1s then removed, and the monolayer is rinsed twice with I'Bs of pH 5.5 to remove any non-adhering bacteria. After the addition of INSTAGEL, the radioactivity of the bound bacteria is measuredin a scintillation counter. (e) Results?

In repeated testswith Compound B, adhesion- increasing mediator proteins were detected in 8 fractions with reproducible igoelectric points.

In treated animals, this mediator-caused increased in adhesiveness was gignificantly reduced compared with the control animals. The results are agsembled in the tollowing table: -& -

td. 26472

TABLE 1

Isoelectric l'oint of the Inhibition of bronchoalveolar lavage Adhesiveness (%) 4.0 13 4.4 65 4.9 65 5.2 61 5.8 39 6.7 67 7.6 79 8.6 ; 72 3. Activity in the in vivo system (a) The bacteria were grown as described above, although without radioactive labelling. (b) Female albino Wistar rats ot female

SPF mice (Chbb:NMR1) were treated with 2 to 200 mg, preferably 20 to 40 mg of benzylamine per kg of body weight or TYLOSE (control) by i.p., i.v., i.m., or p.o. route, preferably by ium. route, 8 to 10 hours before, 15 minutes before and 4 to 5 ’ hours after the infection. The bacterial suspension 8 - (5 x 10 -10 3 bacteria/ml of physiological saline solution) is atomiged in an atomizing chamber at the rate of 0.35 ml per minute for ten minutes.

Alternatively, 10° bacteria in 0.05 ml of physiological saline solution are administered faf . peg? j.n. into the nasal passage with an inhaled anaesthetic, e.g., with chloroform. pach group of animals 1s dissected 1 to 10 hours, preferably 10 hours after the infection, the lungs are removed under steril conditions, and the bacteria are quantitatively determined in homogenized lung by growing on Mclonkey agar.

The bacterial concentration is determined in numbers of baucteria/gm of lung. ; LiaBLi 2

Substance Number of Bacteria/gm _ of Lun ;

TYLOSL 100

Bromhexine hydro- chloride 49

Ambroxol hydro- chloride 31

According to the invention, the compounds of Formula 1 above inhibit colonigation by bacteria such as Actinomyces, pacteroides, Bordetella per- tussis, Corynebacterium diphteriae, Escherichia coli, Haemophilus intluengae, Klebsiella, Myco- bacterium tuberculosis, Mycoplasma pneumoniae, : Nocardia, Yeptococel, Peptostreptococci, Pneumo- cocci, Froteus mirabilis, Fseudomonas aeruginosa;

Serratia, Staphylococci, and Steptococci and tungi such as Aspergillus and Candida.

laf. 26477

The compounds of formula I and the pharma-~ cologically acceptable acid addition salts thereof are non-toxic and therefore well tolerated. Thus, for example, in the mouse : - N-methyl-N-(2-amino-3,5-d1bromo-bensyl )- cyclohexylamine hydrochloride has an

Lbg, of »10 gm/xg p.o., - 8-(2-amino-3, 5-dibromo-benzyl )-trans-4- hydroxy-cyclohexylamine hydrochloride

KE 10 has an Lg, of 2.72 gm/kg p.o., and ~ the other compounds have an LD, of

YL gm/kg p.o.

In view of their inhibiting effect on the adhesiveness of microorganisms, discovered accord- ing to the invention, the compounds of Formula I are sultable for the prophylaxis of infections such +, as infections of the respiratory tract, urogenital tract, mammary glands, akin, and conjunctival membranes, and particularly also for hygiene of the oral and pharyngeal cavities.

To achieve the activity according to the invention, the compounds of Formula I and the pharmacologically acceptable acid addition salts Te thereof with inorganic or organic acids may be administered, optionally in combination with other fad. 26472 active ingredients, preferably substances which =~ additionally guard against infection by microbes, such es sulfonamides or antibiotics, e.g., tetra- cycline, doxycycline, ampicillin, amoxycillin,

cephalexin, or erythromycin, to warm-blooded animals perorally, parenterally, topically, or rectally as active ingredients in customary pre- paration forms suitable for the intended purposes, that is, compuszitions consisting essentially of an inert pharmaceutical carrier and an effective amount ot an inert pharmaceutical carrier and an effective amount of the active ingredient, such as tablets, coated tablets, capsules, granulates, suppositories, solutions, suspensions, emulsions,

ampules, drops, intments, gels, pastes, creams, powders, wound dressings, and sprays.

Advantage- ously the active ingredient or a mixture of dif- ferent active ingredients of Formula I may be administered orally to both humans or animals, in a single dose of trom about 1 to 200 mg (0.013 to

2.67 mg/kg ol body weight), preterably from about © to 150 mg (0.11 to 2.0 mg/kg of body weight), several tives, e.g., from one to four times, daily and externally in a concentration of from about 0.01 to 20 mg per ml or gm of prepara- tion.

A daily dose is therefore trom about 1 to $00 wg (from about 0.013 to 10.7 mg/kg of body weight), preferably from about © to 600 mg (from fuf. 2477 about 0.11 to ..0 mg/kg of body weight). Depending upon the type and body weight of the patient to be treated, on the type and severity of the afflict- ion, on the type of preparation, and on the route of administration as well as on the period or interval over which the administration takes place, it may, however, be necessary to deviate from the

Co above dosages. Thus, it may sufficient in some cases to administer less than the above-mentioned amounts of active ingredient and in some cases the amountg must be exceeded. I'he optimum dosage and route of administratipn of the active ingredients which are necessary in each case can easily be determined by one skilled in the art.

The following examples are intended to illustrate the invention and should not be construed ap limiting the invention thereto.

Jud 76972

SA AHP LLES

Example 1

Tablets Containing 8 mg of Bromhexine lydrochloride gach tavlet has the tollowlng composition:

Component Amount (mg)

Bromhexine hydrochloride ......... 5.0

LaCtOB@ +.veeassoosassoaorosnsnons 50.0

Fotabto Starch ..eecececvacsseesnse 49.9

Polyvinyl pyrrolidone .......ececceo 2.0 :

Magnesium stearate ......coceocees 1.0 t TOTAL: 110.0

Method of Preparation: ‘he active substance is mixed with lactose and potato starch, and the resulting mixture is moistened evenly with an ajueous polyvinyl pyrroli- done solution. {he moistened mass is passed through a 1.5 mm screen, and the granulate is dried at about 40°C in a circulating air dryer and then passed through a screen with a mesh size of 1.0 mm, After the magnesium ateafate lubricant has been added, the tablets are compressed. . Tunch: 7 mm, biplanar, faceted on both gides, dividing notch on one side weight of tablet: 110 mg.

‘ / fof 26475

Example 2

Tablets Containing 60 mg of Ambroxol Hydrochloride sach tablet has the following composition:

Component Amount (mg)

Ambroxol hydrochloride .....ccc.00 60.0

LACLOBE +vvessssroeronanossssvoncen 50.0

Potato SBLATCh sevice eesrsaccvecrene 47.00

Polyvinyl pyrrolidone ............ 2.0

Magnesium stearate .....ccoveeeene 1.0 , IuTab: 160.0

Method of Preparation:

Ine tablets are prepared using a procedure analogous to that of rxumple 1.

Yunch: & mm, biplenar, faceted on both gides, dividing notch on one pide weight of tablet: 160 mg.

Example J . Ampules tontaining o Mg oi Bromhexine ilydrozhkloride “Each ampule has the following contents:

Component Amount

Bromhexine hydrochloride ...ccceccses 8.0 mg

Tartaric acid ..ovecerocecscccscrocne 4.0 ng

Glucogne PIN § LORY me ’ water for Injection .....ccees QeBe ad 4,0 ml tad. 247%

Method of Preparation (100 liters):

The corresponding Juantities of tartaric acid and bromhexine hydrochloride are dissolved in 90 liters of warm water at about 80°C under e current of ti, and with stirring. After cooling to ambient temperature under a current of Ko» the + lucose ig dissolved, and the mixture ig then made up to 100 liters. After sterile filtration and decantation into 2 wl ampules, the ampules are sterilized lor 20 minutes at 120°C. rpxample 4 .

Ampules containing 13 Mg of Ambroxol Hydrochloride wach ampule has the following contents:

Component Amount (1) Ambroxol h,drochloride ....cecce 15.00 mg (2) Citric BCLA eeeoeanesasereeerrt 1.83 mg (3) Sodium piphosphate ceeeeecercr” 7.24 mg (4) Sodium chloride soeesescosereers 14.40 mg } water for injection «..-- q.s. ad 2,00 ml

Method of preparation:

Components (2), (1), 3), and (4) are successively dissolved in heated water. After cooling to ambient temperature under a current of

Ny» the mixture 18 nade up to 100 liters. The resulting mixture is filtered gterile and transferred into 2 ml amjules, and the amnpules are. then

Co sterilized for 20 minutes at 120°C.

Example 5

Syrup tontaining 1.6 mg/ml of Bromhexipe Hydrochloride

One hundred viilliliters of syrup has the 9 following composition: component Amount

Bromhexine hydrochloride ,....... 0.16 gm

Citric acid ¢ * 2 0 0 es Pee ers eee bso 0.50 £m

Sodium hydroxide Je..eeeesesesces 0.04 gm

Sodium DENZOALE + env eennneronnes 0.20 gm

Glycerol ..eievecececosesansnoens 20.00 gm 70% sortitol solution .......¢s.. 70.00 gm

LEhanol cee eeeaseroeesssanncancas 5.00 gm

Haphthol red 5 ...eeeecorccrconcs 0.0003 gm

Plum 11lavoring ..eceeecececasosss 0.20 gm

Distilled water ...........4.8.ad 100.00 ml

Method of lreparation:

The distilled water is heated to 80°¢c, and then the citric acid, sodium nydroxide, bromhexine hydrochloride, glycerol, and sorbitol solution are added successively with stirring and dissolved therein. After cooling to ambient temperature, the sodium beugzoate and naphthol red 5 are added - 17 =

Pd 2047> and dissolved, and the solution of the flavoring in the given quantity of =lcohol is added thereto.

Example 6

Capsules Containing 12 mg oi bromhexine Hydrochloride and 250 mg oi letracycline i ]

Each capsule has the tollowing contents

Component Amount (mg)

Bromhexine hydrochloride ....... 12.0

Tetracycline ....ceceovncernnens 250.0

Dried corn starch ....ceceosnaos 56.0

Magnesium stearate ........oc00. 2,9

TOTAL: 320.0

Method of Preparation: . The bromhexine hydrochloride is triturated with the corn starco to form a homogeneous powder, which is passea through a screen with a mesh size ot 0.3% mm. ietracycline ie wdded to the bromhexine hydrochloride mixture, which is again screened.

Aiter the addition of magnesium stearate, the resulting mixture is homogeneously mixed in a suit- able apparatus, Size 1 capsules are filled with the finished mixture in a suitable capsule-filling machine.

Jof- F477

Lxample 7

Supp

Suppositories for Children Containing B mg of

Bromhexine Hydrochloride _ bach suppository has the following composition:

Component Amount (gm)

Bromhexine hydrochlieride ..... 0.0080

Iolyethyleneglycol 1500 ...... 0.3236

Polyethyleneglycol 6000 ...... 0.2726

Polysorbate 60 (TWLEN ® 60).. 0.4850 : 10 Disodium biphosphate dihydrate 0.0106

TOTAL: 1.10

Method of lreparation:

The bromhexine hydrochloride and disodium biphosphate are ground separately. Lhe polysorbate 60 18 heated, and then the disodium biphosphate dihydrate and bromhexine hydrochloride are sus- pended therein, one after the other,

The susiension of active substance 1s added to the melt of the two polyethylene glycols, which has been brought to the same temperature, and this final mixture is homogenized before : being poured into chilled suppository molds. weight of suppository: 1.1 gm. ’

. / ful. p6072 ’ Lxample 8

Ambroxol Hydrochloride 5olution for Inhalation

Containing 7.5 mg/ml _ _ _ _

One hundred milliliters of solution has the following composition:

Component Amount “mbroxol hydrochloride ...ceoven 0.75 gm 85% ELlycerol ...eceseecesacssvse 5.00 gm

Prim. sodium phosphate ....vce0. 0.60 gm ‘

Sec. sodium phosphate .s...eeeese 0.055 gm

BENzOLC ACLA eveeevnreennneasses 0.10 gm

Distilled water ........ q.8, ad 100,00 ml

Method of Preparation:

The 854 glycerol, prim, and sec. sodium . phosphate, aud benzoic acid are dissolved until clear in distilled water at 50° to 55°C. After cooling to 25°¢, the ambroxol hydrochloride is dissolved therein, and the mixture is then filtered. sxample Y wound Rinse Solution containing 10 mg/ml of

Ambroxol Hydrochloride

One hundred milliliters of solution has the tollowing composition:

Component Amount

Ambroxol hydrochloride ......e000 1.00 gm 70% sorbitol solution .......... 2.00 gm

Citric acid. H,0 oe * ss ee tas esas 0.40 gm

Sodium hydroxide ...cecevocevncs 0.23 gm

Shdium chloride sev. ceriecvienne 0.10 gm

Benzallkonium chloride ......e000 0.025 gm

Distilled water ....... q.8. ad 100.0 ml fiethod of Preparation:

The 704 sorbitol solution, citric acid, sodiwn chloride, and benzalkonium chioride are I dissolved until clear, with stirring, in distilled water. After a completely clear solution is obtained, sodlum hydroxide is added and dissolved therein. pt = 6.2; lsotonic.

Lxample 10

Ointment Containing 4 mg/gm of Bromhexine Hydro- chloride and 200 mg/gm of byythromyein

One hundred grams of ointment has the following composition:

fal. 6472

Component Amount (gm)

Bromhexine hydrochloride ........ 0.40

Erythromycin ...ceeeeeeecececcceee 20.00

Cetyl alcohol ..eeeevessocveacnns 0.50

WOOL WAX veeiorosasssacsssconsonse 3.45

Thin paraffin oll .....cevvceenn. 13,60 white vaseline ......ceeuvoncenns 62.25

TOTAL: 100.00

Method of Preparation:

At 60°C the cetyl alcohor, wool wax, thin paraftin oil, and white vuseline are inelted, tiltered, and cooled to about 45°C. At this temperature the bese is still liyuid. Lhe active substances bromhexine hydrochloride and erythromycin are iv thopoughly incorporated therein, the mass is stirred until cold, and air 1s removed, if necessary.

Example 11

Mastlitis Cintment for Animals tontaining 12 mg/gm of Ambroxol hyd drochloride and »0 mg/gm Tetracycline

One hundred grams of ointment has the following composition:

Jol. 26472 ~ vomponent Amount (gm)

Ambroxol hydrochloride ......... 1.20

Tetracycline ....oieeccececoccos 5.00 white vaseline a 25.15

WOOL WAX yecsecevoasesseaesonerac 2.75 lsopropylmyristate ....coceenee 32.95

Thick parafiin oil ..e.ecececees 19.50

Thin paraffin oil ..eceeeeecoces 13.45

ToT ab: 100.00

Method of Preparation:

At 50°¢ white, vaseline, wool wax, iso- propylinyriestate, and thick and thin paraffin oils are melted and thoroughly mixed. Ihe ambroxol hydrochloride and tetracycline are then homo- / genized therein, and the mix.ure is cooled with gtirring.

Example 12

Capsules containing 12 wg of N-(3,5-Dibromo-2- hydroxy -benzyl)-trans-4-hydroxy-cyclohexylanine llydrochioride and 250 mg of tetracycline gach capsule has the following contents:

Component Amount (mg)

Active subStAnNCe «eeeeececoee 12.0

Tetracycline es.eeeeseecvocones 250.0 /25 Dried corn starch ececeeeeeeeee 56.0

Magnesium gtearate ....eseroe _ 2.0 \ TOrAL: 320.0

If. 96477

Method of lreparation:

The capsules are prepared using a procedure analogous to that of Example 6.

Lxample 12

Tablets tor Sucking Containing 30 mg of Ambroxol

Hiydrochloride rach tavlel has the iollowing composition: vowponent fmount (mg)

Ambroxol hydrochloride ......... 30.0

Sorbitol, suitable for tablet )

MBKINE eeecvsecscsccosaarsdoe 666.5

Magnesium stearate cieeeosceasene 349

LOLAL: 700.0

Method of Preparation:

The active substance is homogeneously mixed with Lhe same yuantity oi sorbitol, the remaining components are added, ana again a homogeneous mixture is prepared (mixture ready lor compressing).

This is compressed in a suitable tablet-making machine to fora tablets. weight: 700 mg

Diameter: 13 mm, radius of curvature 16 mm

Appearance: white, round, biconvex.

Ss fut 26477

Example 14

Tablets for Sucking Containing 8 mg of Bromhexine liydrochloride .

Each tablet has the following composition:

Component Amount (mg)

Bromhexine hydrochloride «....... 8.0

Sorbitol, suitable for tablet

MAKLINE seessoeasensassoerecsos 686, 5

Magnesium stearate ......ccccv00es __ 3.9

TOTAL: 700.0

Method of Preparation: .

The tablets are prepared using a procedure analogous to that of Example 13.

Lxample 15

Film-coated Tablets Containing 60 mg of Ambroxol liydrochloride and 500 mg of Cephalexin kach tablet core has the following composition: component Amount (ag) (1) Ambroxo) nydrochloride .........c... 60.0 (2) Cephalexin ..oeceeececconcenecenrees 500.0 (3) LactoBe ...cesessescsconcoraacoconee 70.0 (4) Dried corn starch (...oeevvececenene 80.0 (5) Polyvinyl pyrrolidone .............. 16.0 (6) Magnesium stearate .....ceeeecevocns 4.0 __

TOTAL: 7230.0 ’

Method of Preparation: components (1), (2), (3), and (4) are mixed together and moistened with a 10% solution of component (5) in 70% ethanol. Lhe moistened mass is passed through a screen with a mesh size of 1.5 mm, dried, and then passed through the same screen again. Component (6) is then added, and tablet cores are produced from the mixture useing a tablet press.

Coating:

A coating consisting of a solution of ethylcellulose in ethanol is applied to the pressed tablet cores. lhe weight is increased by 1.5% by this tilm-coating. weight of tablet core: 730 ing

Punch; 1p mm

Example 16

Dry granulate for Syrup Containing 15 mg of Ambroxdl

Hydrochloride and 250 mg of Cephalexin _

Five milliliters of Syrup has the following

Composition:

Component Amount

Ambroxol nydrochloride ........ce0s 15.0 mg

Cephalexin ...coeveeceereenceannens 250.0 mg

Sodium citrate .....ceeesensacvoans 50.0 mg

Dioctylsodiumsullosuccinate cece 1.0 mg

Polyvinyl pyrrolidone ........eooees 50.0 mg jiagnesium aluminum silicate ....... 10.0 mg

Sodium saccharin ...ceceeiveaccacss 5.0 mg

Sodium salt of methyl p-hydroxy-

DENZOALE seveaseoossosnsoncsancs 6.0 mB

Sodium salt of propyl p-hydroxy-

DENZOALE ceevevsaeroassrocesscne 1.5 mg

Powdered 11lavorifg ...ceeecacccnens 10.0 ng oilicone antitroam agent ....c.e.ene 0.25ng 19 Mannitol ea 6 6 00 8s ss se BOER ON q.8. ad 2.0 gin

Method of reparation: (a) Solution tor granulation

Ihe dioctylsodiumsul fosuccinate, silicone antifoamer, and some of the polyvinyl pyrrolidone are dissolved ar dispersed in the juantity of water required tor granulation. : (b) Mixing of powders . All the remaining components are home#ien- ously mixed, with the exception of the flavoring.

(¢) Granulation

Ihe mixture of powders is moistened with the liquid for granulating. lhe moist mass is screened to a maximum 2 mm, dried, and screened again. (d) Final filixture

The ilavoring is adued to the granulate and homogeneously distributed. forty milligrams of granulate plus 7b ml oi water yield about 100 . mL of syrup ready lor use.

Five milliliters ol syrup, which is equal to one dose, contain 15 mg of ambroxol hydrochloride and 250 mg ol cephalexin. example 17 1% Dry Granulate ror urops Containing 15 mg/ml of

Ambroxol liydrochioride and 250 mg/ml of Cephalexin

One milliliter of drop suspension has the following composition:

$2 {

Component Amount

Ambroxol hydrochloride ........... 15.0 mg

Cephalexin .eeeieeceiracrnacecces 250.0 ng

Sodium citrate .......i cies 30.0 mg 9 Methyl celluloBSe ..ccociseviceaans 2.0 mg tolyethyleneoxide ....cveceveranns 2.0 mg

Sodium salt oi sethyl p-hydroxy-

DEeNZOBLE sei eeeesrnsansccassnoss 1.2 mg

Sodium salt ot prop,l p-hydroxy- benzoate .ieeeevesrssrscsnssonns 0.3% mg

Powdered flavoring ......coceseses 3.0 mg

Silicone anltilo. mung agent ....... 0.05mg

Powdered sugar eer ies. g.8..8d 0.404gm ilethod of Preparation:

Solution for greunulating:

Polyethylene oxide, silicone antifoamer, and some of the methyl cellulcse are dissolved or dispersed in the juancity of waler required for sranulation. the remainder of the process lis as in wxample 16. rorty grams of granulate plus 75 ml of water yleld about 100 ml of drop suspension ready {or use. x, upe iillliiter of drop suspension, which is eyual to one dos?, contadns 15 ng ol ambroxol hydro- chloride and 290 wg of cephaiexin.

Id. starr -

Example 18

Oblong Tablets Containing 30 mg of Ambroxol Hydro- chioride and 500 mg_of Ampicillin wach tablet has the following composition:

Component Amount (mg) (1) Ambroxol hydrochioride ......... 30.0 (2) Ampicillin cieeaceriecerovocecnes 500.00 (3) Powdered Lactose ....ecveccracons 365.0 (4) Dried corn Starch .....cceeeoeee 180.0 (5) Polyvinyl pyrrolidone .ces.eeeane 21.0 (6) Magnesium stearate Chee scnanans 4.0

TOTAL: 1100.0

Method of Preparation:

Components (1), (2), (3), and (4) are mixed together and moistened with a 103 edlution of com- ponent (5) in 7k ethonol. LIne moist mass is further treated as in vxample 15. weight ot tublet: 1100mg

Mold: 8 x 17 mm oblong. rxample 1) ‘ Ampules Containing 6 mg/ml of Ambroxol Hydrochloride . and 100 mg/mi of impicillin a

The ampule solvent has he following composition:

Conpnosnk Mente }

Ambyrizo] hvAr~=hl>rids . . ... ial; BRETT

S~diwm hydeoxids =aluticn Lo. 2.5 ng

Tortaric acid oe oa 5.7 mg

Polvethylena oxide oo... oo TR.O wg

Water for injection oo... rl Loong

Dry ampule:

Sodium ampicillin: 00.90 mg

Mathod of Preparation:

Thie ambroxol hydrochloride and tartaric acid are dissolved with stirring. the prlyethylene oxide is dissolved, and the solution is adjusted to a pH of 4.9 with sodium hydroride aolntion. It is filtered sterile through membrane filters, 156 transferred into ampules, and gtertlized for ten minutes at 120°C.

Example 29

Ampules Containing 156 mg/ml of Ambroxol Hydro- chloride and 20 me/ml of Noxvave | ing llvdrochloride

Each ampula has the following contents: ar

BAD ORIGINAL

- ee

Component Amount.

Doxycycline hydrochloride ...... 100.00 ny

Ambroxel hydrochloride ..._._... 75.0 mg

Magneaivm oxide oo... LL... 9.1 mg

Sodium pyrosulphite ............ HhH9o0.9 mg

Polyethylene oxide ......._...... 500.©® mg

Water for injection ...._.gq.s8. ad Rh. ml

Method of Preparation:

The substances sre dissolved succensively in water and filtered through membrane filters. The solution is transferred into ampules under a current. ot nitrogen. The solution cannot be sterilized.

Example 2)

Capsules Containing 150 mg of Ambroxol Hydrochloride and 109 me of Dozyevellve

Each capsule bar Lhe following contents:

BAD ORIGINAL edd -

(uf. 26472

Component Amount. (mg) 40X spray pellets, consisting of wax mixture and active substance¥ .. 375.0

Btarch .....coeccccacaacaaacneneaenanan- ho.0

Lactose .......... ccouecanacacnacanonns 50.0

Highly dispersed silicone dioxide .... 7.0

Magnesium stearate _.................. aa

TOTAL: 400.0 —

The active substance is suspended in the melt heated to 70°C by means of an uLTRA-TURRAX(B) high frequency mixing device and sprayed in a suitable apparatus. ot

Method of Preparation: 16 The mixture is transferred into size © long gelatine capsules, in a suitable capsule-making machine equipped with a tablet insertion station, together with a coated 6 mm core containing 100 mg of doxycyline and conventional tablet-making excipients.

Woight of capsule: 490 mg.

Example 22

Tablets Containing 60 mg of Ambroxol Hydrochloride and 6800 mg of Ervthromyein

Each tablet has the following composition:

Component Amount. (mz) (1) Ambroxol hydrochloride ...._....... 60.0 (2) Erythromycin ........_.._._.._....... 600.0 (3) Powdered lactose ..._.............. 866.0 (4) Dried corn starch .._..__.._.._......... 265.0 (6) Polyvinyl pyrrolidone ...._.._...... 21.0 (8) Magnesium stearate ......___...... ___ 4.0

TOTAL: 1900.0 ‘

Method of Prepavation:

Components (1),+(2), (3), and (4) are mixed together and moistened with a 10% solution of component (5) in ethancl. The moist mass is further treated as in RKxample 15.

Weight of tablet: 1300 mg 16 Mole: 8.5 x 18 mn oblong.

Example 23

Children’s Tablets Containing 7.5 mg of Ambroxol

Hydrochloride and 1650 mg of Ervthromycio

Rach tablet has the following composition:

Component Amount (mg) (1) Ambroxol hydrochloride ...._......._.. 7.5 (2) Erythromycin ....................... 150.0 oo (3) Powdered lactose ........_.___....... 96.0 - 34 i

/ ful. 260.77 (4) Dried corn starch ..............--.. 68.25 (6) Polyvinyl pyrrolidone .............. 5.2b (6) Magncmsium stearate o.oo... 1.00 _

TOTAL: 3s. 00

Method of Preparation:

Components (1), (2), (3), and (1) are mixed together and moistened with a 19% soluLion of component (b) in ethanol. The moist muss is further treated as in Example 1b. 19 Weight of tablet: 325 mg

Diameter of punch: 19 nm

Example 24 .

Dry Granulale for Drops Containing 15 mg/ml of 156 One milliliter of drop suspension has the following composition:

Liomponenb Amount

Ambroxol hydrochloride ............ 15.9 mg , Erythromycin base equivalent ...... 300.9 mg - 20 SOdiUN CLLEBLE «ners eaneannennaes J0.0 mg

Sodium lauryl sulfate ............. 1.¢ mg

Melby] cellulose ..............a-.- 2.96 ng

Sodium saccharin ... cue anan 1.0 ng

Sodium salt of methyl p hydroxy benzoate . ....-ccc-ecnacacnanann= 1.2 mg — db : .

BAD ORIGINAL

Mined

Sodium salt of propyl p-hydroxy-

BONZOBLES - eee eecinncnnancana- 0.3 wg

Polyvinyl pyrrolidone ............. 3.0 we

Powdered flavoring ............-...- 3.0 mg §1licone antifoamer ............... ©.85 wg

Powdered sugar .......----9.8. ad @.4 gn

Method of Preparation:

The preparation is analogous to that in

Example 17. DPorty grams of granulate and 75 ml of water vield about 100 ml of drop suspension ready for ume. One milliliter of drop suspension, which is equal to one dose, containm 15 mg of ambroxol hydrochloride and 300 mg of erythromycin base equivalent. 156 Example 25

Dry Granulate for Byrup Containing 8 mg/ml of

Five milliliters of syrup have the following compsition:

Component Amount:

Ambroxol hydrochloride ... ......... 390.0 mp

Erythromycin base equivalent ...... 620.0 mg

Sodium citrate . ......cccceceu-.... 100.0 ng

Hydroxyethyl cellulose ............ 2.0 mg ff. 2447

Magnesium aluminum silicate ....... 15.0 mg

Aluminum oxide .................... 15.0 mg

Sodium saccharin .................. 4.0 mg

Sodium salt of methyl p-hydroxy- benzoate ........._............ 8.2 mg

Sodium nalt of propyl p-hydroxy-- benzoate ..........c.oeoorannn 1.5 mg

Powdered flavoring ................ 10.0 mg

Silicone antifoamer ............... 0.15 mg

Powdered sugar ............Q.8. ad 2.00 gm

Method of Preparation:

The preparation is analogous to that iu

Example 136, with the eXcopbion that instead of polyvinyl pyrrolidone, some of the hydroxyethyl cellulose is dissolved in water together with the silicone antifoamer. Forty grams of granulate plus 75 ml of water yleld about 100 ml of syrup ready for use.

Five milliliters of syrup, which is equal to one dosu, contain 30 mg of ambroxol hydrochloride and 600 mg of orythromycin base equivalent.

Example 26

Drops Containing 7.5 mst/ml of Ambroxn). Hydrochloride

One hundred milliliters of drop solution has the following composition:

fod. 21472

Component Amount:

Ambroxol hydrochloride ............ ©.75 gm 85% glycerol fee eee aaa. 5.00 gm

Citric acid monohydrate eee. 0.20 gm

Sodium biphosphate dihydrate ....._. 0.315 gm

Sodium chloride Sree eee. 0.59 gm

Sodium saccharin Chee enema. 0.025 gm

Flavoring Cree n eee e et ieemeeom mean. 0.05 gm

Benzoic acid _......._.._..._. 9.20 gm

Distilled water ceceacr.....q.8. ad 109.00 ml

Mathod of Preparation:

The drop solution in prepared using a procedure analogous to that of Example 25.

Example 27 186 Tablets Containing 15 ml of Ambroxol Hydrochloride

Each tablet has the following composition: ’

Companeunt Amount, (1) Ambroxol hydrochloride crrensracean 165.0 (2) Amoxycillin fee eds aaa. 750.0 (3) Lactose ee ee ee em ee 400.0 (4) Dried corn starch eee aan aaa. 9305.0 (5) Polyvinyl pyrrolidone eee eee 265.0 (8) Magnesium stearate ee em aa 5.0

TOTAL: 1500.0 rl (ol 26472

Method of Preparation:

Components (1), (2), (3), and (4) are mixed together and moistened with a 10% solution of component (5) in ethanol. The moist mass is further treated analogously to the procedure of

Example 15.

Weight of tablet: 1500 ng

Mcld: f! x 17 mm, oblong.

The examples above illustrate several pharmaceutical compositions comprising a compound of the present invention as sn active ingredient and represent the hesl modes contemplated of using the oo invention. It should be understood that the active o substance present in each pharmaceutical composition could be replaced by onc or wore other compounds of

Formula I.

The preceding specific embodiments are 11lustrative of tho practice of the invention. It is to be understood, however, that other expedients known to those skilled in the art or disclosed herein, may be employed without departing from the spirit of the invention or tha scope of the appended claims.

Claims (1)

1. CLAIMS: : . :

   l. A method for the prophylaxis or treatment of . infections in a warm-blooded host in need of such treatment which comprises perorally, parenterally, rectally or topically administer- ing to said host an efiective amount of at least one compound of the formula (I). Par B CH, ~ N Rs 2 ~N Pp Ry 1 TT Br wherein Ry represents a hydooxyl group in the 2- or 4-position or an amino group in the 2- position; R, represents a hydrogen atom or a methyl group; and Ry represents a cyclohexyl or hydrocyclo- hexyl group, or a pharmacelogically accept- able acid addition salt thereof with an inorganic or organic acid.

   fal. 2by.72

   2. The method of claim 1, wherein the compound is a compound in which Ro and Rg together with the nitrogen atom between them represent an N-methyl- cyclohexylamino or cis-3--hydroxy--cyclohexylamino Broup, or a pharmacologically acceptable acid addition salt thereof.

   3. The method of claim 1, whorein the compound is N-mathyl-N-(2-amino--3.5 -dibromo-benzyl )-cyclo- hexyvlamine, or a vharmacologically acceptable 1e acid addition salt thereof.

   4. The method of claim 1, wherein the compound is N-(2-amino -3,5-dibromo-benzyl )-trans—4-hydroxy— cyclohexylamine, or a pharmacologically aoccept- able acid addition salt thereof. 156 6. The method of claim 1, wherein the compound is N-(3,5-djbromn—-4-hydrosy-benzyl )-cis-3-hydroxy—- cyclohexylamine, or 5 pharmacologically accept- able acid addition malt thereof.

   8. The method of claim 1, wherein the compound is N-(3,65-dibromo-2-hydroxy—-benzyl )--trans--4- hydroxycyclohexylamine, or a pharmacologically acceptable acid addition salt thereof.

   7. The method of claim 1 which comprises adminis- toring from about 1 to about 200 mg of a com--

   Jaf. 2477 pound of formula I or a pharmacologically acceptable acld addition salt thereof.

   8. The method of claim 1 which comprises topic- ally applying to a host a preparation contain- ing from about 9.01 to 20 mg/ml or gm of a compound of formula I or a pharmacologically ’ acceptable acid addition salt thereof.

   9. The method of claim 1 which comprises administer— ing an effective amount of a compound of formula J or a pharmacologically acceptable salt thereof and an antibiotic. oo

   10. The method of claim 1, wherein the infections are infections of the respiratory tract, the urogenital tract, the mammary glands, the skin, the conjuctiva, or the oral or pharyngeal Te cavities. CHRISTIAN KRUGER —- ANGELIKA MARIA ERNE ROLF-GUNTHER WERNER HANS GOETH : HAZARD ZIEGLER Inventors