HU226845B1 - Process for the preparation of an injection formulation comprising tuberculin as agent - Google Patents

Abstract

The injection preparation comprises purified freeze-dried tuberculin (1-50 TE/ml) and liquid vehicle, and optionally contains additives. The vehicle is sterilized distilled water, normal saline solution or lidocaine (0.5-1 mass/mass%). ACTIVITY : Dermatological; Virucide. No biological data given. MECHANISM OF ACTION : None given.

Description

The present invention relates to a process for the preparation of an injection composition containing the active ingredient tuberculin.

The treatment and treatment of warts caused by human papillomavirus (HPV) is still a major problem for dermatologists [Jablonska S., Majewski S., Obalek S., Orth G., Cutaneous warts, Clin. Dermatol. 1997, 15 (3): 309-19.].

Warts are predominantly located on the hands and feet, but in some cases atypical localization may occur. Particularly problematic is the fact that a very large number of warts are located on the same area of skin. Without adequate immunity against HPV, there is a high risk of recurrence, whichever method of treatment is used.

Physical removal of viral warts is the primary treatment option. The most common method is freezing with liquid nitrogen [Conolly M., Bazmi K., O'Connell M., Lyons J.F., Bourke J.F. Cryotherapy of viral warts: sustained 10 freeze is more effective than the conventional method, Br. J. Dermatol. 2001, 145 (4) 554-7.].

In this method, a blast forms a blister due to the sudden cold and if done properly, that is, for a sufficiently long time to freeze, a deep freezing effect results in a second degree frostbite and the resulting blister lid contains the vast majority of virus particles.

The disadvantage of this method is that it is quite painful and causes a lot of discomfort, as it takes several days or even weeks for the wound to heal to be removed after removal of the bladder lid.

It is more effective when, after a brief local freeze or after local anesthesia, a Volkmann canal or electrocauter is used to mechanically remove the wart [Jablonska S., Majewski S., Obalek S., Orth G., Cutaneous warts, Clin. Dermatol. 1997, 15 (3): 309-19.].

This procedure is less painful due to local anesthesia, but here too, wait at least one week for proper wound healing and before the affected part of the body is spared. This can often lead to partial disability during recovery.

Local irritants and sensitizers, such as dinitrochlorobenzene (DNCB) or squaric acid, can very often lead to an extremely pronounced inflammatory reaction [Buckley, DA, Du Vivier, AW: The Therapeutic Use of Topical Contact Sensitizers in Benign Dermatol]. . 2001, 145 (3): 385-405.].

A further disadvantage of the procedure is that DNCB, as a carcinogen, is no longer applicable for human use in many countries.

Numerous contradictory studies have reported the use of the preferred hydrogen receptor blocker cimetidine in the treatment of group-wise, therapeutically resistant warts (Yilmaz, E., Alpsoy, E., Basaran, E. Cimetidine therapy tor warts: a placebo-controlled, double-blind study, J. Am. Acad. Dermatol. 1996, 34 (6) 1005-7.].

Similarly, for treatment-resistant, a large number of warts predominantly located on the foot may be given oral acitretin or previously etretinate (Boyle, J., Dick, D.C., MacKie, R.M.) Treatment of extensive viral warts with etretinate (Tigason) in a patient with sarcoidosis. Clin Exp Dermatol 1983, 8 (1): 33-6.].

Due to its many side effects (pronounced palpation of the palms and soles, hair loss, dryness of the skin and lips, sweating, nail plate growth) and its ineffective therapeutic effect, this product is rarely chosen.

Local immunotherapy is quite difficult to carry out and the costs are high.

Topical interferon preparations [Gross G., Rogozinski T., Schofer H., Jablonska S., Roussaki A., Wohr C., Brzoska J.: Recombinant interferon beta gel as an adjuvant in the treatment of recurrent genital warts: results of a placebo-controlled double-blind study in 120 patients. Dermatology 1998, 196 (3): 330-4.] And, more recently, imiquimod-containing exteriors (Hengge UR, Esser S., Schulteworter T., Behrendt C., Meyer T., Stockfleth E., Goos M.). : Self-administered topical 5% imiquimod treatment of common warts and molluscum contagiosum (Br J Dermatol 2000, 143 (5): 1026-31.).

Bacillus-CalmétteGuerin (BCG) vaccination is mandatory in Hungary, which increases the body's immunity to Mycobacterium tuberculosis. IV. type of immunity develops. Its potency can be measured at mandatory times or in the case of disease using the so-called Mantoux test (tuberculin test) under standard conditions [Mazurek GH, LoBue PA, Daley CL, Barnardo J., Lardizabal AA, Bishai WR, lademarco MF, Rothel JS: Comparison of a whole-blood interferon gamma assay with tuberculin skin testing tor detecting latent Mycobacterium tuberculosis infection. JAMA 2001,286 (14): 1740].

During the test, highly purified cell wall antigens of mycobacterium tuberculosis hominis and bovis are inoculated intracutaneously. The method is one of the most reliable in the IV. type of immunity.

However, a tubular injection of tuberculin has not yet been reported for use in the treatment of skin disorders caused by human papilloma, such as warts, by direct injection into the wart.

WO9955347 proposes treatment with mycobacterium, BCG, to treat infection by human papillomaviruses.

W00048615 proposes the use of Mycobacterium vaccae for the treatment of viral infections.

US 4657761 describes the use of an injection formulation containing combined vaccines by subcutaneous administration.

The disadvantages of conventional formulations and methods of treating warts are as follows.

HU 226 845 Β1

The most common mechanical interventions are: long-lasting, harder to heal wounds, which are painful and significantly affect the way you feel and work.

Serious side effects (dryness of the eyes and skin, hair loss, liver dysfunction) may occur if systemic vitamin A derivatives are used.

In case of topical immunotherapy, pronounced inflammation may occur.

It can be stated that the efficacy of the known compositions and their application is rather low.

It is an object of the present invention to provide a formulation for injection which is highly effective in the treatment of warts caused by human papillomaviruses by topical application, i.e., by injection directly into the wart.

The present invention therefore relates to a process for the preparation of an injection composition containing the active ingredient tuberculin.

The method is characterized in that the tuberculin active ingredient, the liquid carrier and optionally additives are prepared in a manner known per se to formulate an injection preparation for the topical treatment of papillomavirus warts.

The injection preparation according to the invention contains 1-50 TE (tuberculin unit) of purified, preferably lyophilized, tuberculin.

Preferably, the composition contains 25 TE purified lyophilized tuberculin.

Tuberculin, also known as PPD, is a purified and selected protein derivative of the Mycobacterium bacterium strain.

TE (1 unit of tuberculin) corresponds to 20 ng of PPD.

This definition can be found in Kuwawara S .: J. Bioi. Chem. 250, p. 2556-2562 (1975).

Preferably, the composition of the invention comprises sterile distilled water as the liquid carrier.

In another embodiment, the composition prepared comprises a physiological saline solution or a sterile 0.5 to 1% solution of lidocaine as a liquid carrier.

The composition of the invention may contain as an additive magnesium sulfate or calcium sulfate or a mixture of magnesium sulfate and calcium sulfate.

The advantage of using the composition according to the invention over the methods used so far is as follows.

The composition according to the invention is administered by injection and the stinging only causes very slight pain and no wound is formed.

After the procedure, no bandage is required and no precautions are taken, with mild redness occurring only 24-72 hours after the prick.

The resulting composition is applied topically to the wart.

On average, studies in groups of patients require two injections with an interval of three weeks, and 82% of the treated warts disappear completely during treatment. And 57% of non-responding warts disappear completely after the third treatment.

The preparation according to the invention and its use are illustrated by the following examples.

Example 1

Purified tuberculin is prepared from a filtrate of cultures of human and bovine Mycobacterium tuberculosis strains grown on Sauton's medium by purification.

The solution thus obtained is lyophilized.

Dissolve 25 TE of lyophilized purified tuberculin in 1 ml of distilled water.

The mixture is homogenized and ampouled.

Example 2

The procedure of Example 1 was followed, but sterile physiological saline was used instead of distilled water.

Example 3

Example 1, but using 1% sterile lidocaine solution instead of distilled water.

Example 4

The procedure of Example 1 was followed, however, using a solution containing 10 mg / ml of magnesium sulfate instead of distilled water.

Example 5

The same procedure as in Example 4 was used, but instead of distilled water, a solution containing 10 mg / ml of calcium sulfate was used.

Example 6

The same procedure as in Example 4 was used, but instead of distilled water, a solution containing 10 mg / ml of a mixture of magnesium sulfate and calcium sulfate was used.

Example 7

The procedure of Example 2 is followed, however, with a mixture of magnesium sulfate and calcium sulfate at a concentration of 2 mg / ml in sterile physiological saline.

Example 8

EXAMPLE OF EFFECTS Different groups of patients are treated partly with the injection composition according to the invention and partly with conventional liquid nitrogen freezing. Partially solvent-only injection.

The treatment with the composition of the invention was carried out using a novel method, which is unconventional and which is as follows.

The area to be treated is photographed. The area to be treated is then wiped with a 70% alcoholic sterile gauze pad. The wart is injected with 0.1 ml of the injectable preparation of the invention or as a solvent alone control.

The area is wiped again with an alcohol gauze pad.

HU 226 845 Β1

A follow-up examination will take place after 3 weeks, during which the first treatment will be followed by inoculation of the warts that have not disappeared, followed by a further photograph of the area.

Further control examinations are performed after 3 weeks, at which time warts that have not disappeared from the previous treatment are re-injected and photographic documentation is prepared.

The conventional treatment is as follows.

Warts formed on the scalp are treated with liquid nitrogen by applying a cotton ball to the liquid nitrogen and treating the warts and the surrounding 1 mm intact area with a squeeze for 15 seconds on each wart. The bladder formed after the treatment was removed the next day and the wound was treated with 3% hydrogen peroxide brush and covered with boric acid powder.

For warts that have not disappeared, the treatment is repeated after 3 weeks as above.

Warts on the soles are also treated with liquid nitrogen as described above. The bladder formed after treatment was removed the next day and the wound was treated with 3% hydrogen peroxide brush and covered with boric acid powder. Criteria excluding treatment with a composition of the invention: generalized dermatitis, a history of allergic reaction to tuberculin vaccine or lidocaine, immunosuppressive status, pregnancy, breastfeeding.

Patient groups:

I. Treatment of warts on hands:

For warts on the hands, 52 patients (24 males and 28 females) were examined.

The average age is 17.1 years (5-32 years).

The mean age of participants in conventional (freezing) treatment was 15.4 years (20 persons), while the mean age of patients receiving the lyophilisate diluted in sterile distilled water was 18.7 years (16 persons) and the saline diluted in saline was 16 years. those with an average age of 16.7 years.

The number of warts per capita was 3.4 in the conventional group, compared with 5.2 in the immunotherapy group treated with the lyophilisate diluted in distilled aqueous solution and 5.5 in the saline lyophilisate treated with the saline diluent. number of warts.

For plantar warts, 65 patients (32 males and 33 females) were enrolled. Age average is 24.5 years (9-51 years).

The average age of participants in conventional freezing treatment was 20.9 years (19 persons), while in the group receiving immunotherapy, distilled water diluent and treatment with magnesium sulphate and calcium sulphate (10 mg / ml of each compound were dissolved in the solvent) ) 29.6 years (12 persons) 27.6 years (13 persons) in saline diluent and cases treated with magnesium sulphate and calcium sulphate as compared to lyophilisate diluted in 0.5% lidocaine solution The average age was 24.1 years (21 persons).

In the placebo (saline) alone group, the mean age was 20.5 years (8 persons).

The average number of warts per capita in the conventional freeze treatment group was 6.9 warts / person physiological saline diluent and in the immunotherapy group treated with distilled water diluent and treated with magnesium sulfate and calcium sulfate. The number of warts per capita was 6.9 in the group treated with the magnesium sulfate and calcium sulfate additive and 7.1 in the group treated with the lyophilisate diluted with 0.5% lidocaine solution.

The mean number of warts was 3.7 per person in the placebo group, ie, physiological saline.

There are 7 different groups during treatment:

Group 1: Freezing of warts on the head

Group 2: Immunotherapy of warts on the head with 25 TE / ml in sterile distilled water

Group 3: Immunotherapy of cervical warts with 25 TE / ml in sterile physiological saline

Group 4: Freezing of warts on soles

Group 5: Immunotherapy of plantar warts with 25 TE / ml of tuberculin dissolved in sterile distilled water and supplemented with 10 mg / ml of magnesium sulphate and 10 mg / ml of calcium sulphate.

Group 6: Immunotherapy of plantar warts with 25 TE / ml of tuberculin dissolved in sterile physiological saline and supplemented with 10 mg / ml of magnesium sulphate and 10 mg / ml of calcium sulphate.

Group 7: Immunotherapy of plantar warts with 25 TE / ml tuberculin diluted in 0.5% lidocaine solution.

Group 8: Treatment of plantar warts with sterile 0.9% saline.

The results of the treatments are as follows.

Of the patients undergoing conventional treatment of palpable warts, three patients did not complete the study because of the pronounced side effects because of the extreme post-freezing burning sensation and blistering. Everyone underwent immunotherapy until the end of the study.

At the visit 3 weeks after the first freeze or injection, the average number of patients per patient was 2.5 after freezing (group 1), after the first immunotherapy in group 2 1.9 and in group 3 , Was 0 pieces.

At the second control trial, that is, after another three weeks, the mean number of warts after freezing was 2.0, compared with 0.4 for Immunotherapy and 0.5 for Group 3.

Among the patients involved in the traditional treatment of warts on the soles, the side effect is pronounced4

Because of many, we did not complete the study in five patients because of the pronounced post-freezing burning sensation, blistering, and gait difficulty.

All patients underwent immunotherapy until the end of the study. 5

At the visit three weeks after the first freeze or injection, the mean number of patients was 2.5 after freezing (group 4) and 3.0 after the first immunotherapy in groups 5 and 6. 10

In group 7, the mean number of patients was 3.2.

In group 8, the average number of warts remained 3.7. At the second control trial, that is, after another three weeks, the mean number of warts after freezing was 2.3 (Group 4), compared with 0.7 for Immunotherapy and 0.5 for Group 6. in group 7 it was 0.8. In Group 8, the average number of warts was 3.5. 20

Sixteen weeks after the first treatment, the number of warts was re-examined and each group, except for the freezing procedure, showed further improvement and steady-state results.

The results are summarized in Table I. 25

Side effects:

During the treatment of warts on the scalp, three patients in the freezing group developed superficial wounds. Three patients who received immunotherapy had more severe redness, which required overnight application of icing. In warts on the soles, one patient developed a wound superfusion after freezing, whereas in immunotherapy, only one patient reported a burning sensation at 48 hours after stinging, which ceased to occur after icing.

The complaints of patients in both groups, whether palm or plantar, were significantly different.

The burning sensation during and after freezing was extremely unpleasant and made it difficult to bear the blisters on their hands because it greatly hindered them in their work and daily activities. Freezing on the soles caused similar discomfort.

In contrast, the groups treated with the composition of the invention did not develop similar complaints except for the small number of side effects described above. No adverse reactions were observed in patients treated with saline only.

Table I

Number of groups 1 2 3 4 5 6 7 8 Mean number of warts before treatment 3.4 5.2 5.5 2.9 6.7 6.9 7.1 3.7 Mean number of warts after 1st treatment 2.5 1.9 2.0 2.5 3.0 3.0 3.2 3.7 Mean number of warts after 2nd treatment 2.0 0.4 0.5 2.3 0.7 0.5 0.8 3.5 Average number of warts 16 weeks after first treatment 2.2 0.4 0.4 2.1 0.6 0.5 0.6 3.5

From the above results, it is apparent that the treatment with the composition of the invention 40, although involving a small puncture, is still much more advantageous than the freeze method most traditionally used.

The composition of the present invention is more effective in treating viral warts and has a much narrower and less severe spectrum of side effects.

Claims (8)

PATENT CLAIMS CLAIMS 1. A method of preparing an injection formulation containing the active ingredient tuberculin, comprising mixing a tuberculin, a liquid carrier and optionally an additive prepared and known in the art to form an injection formulation for topical treatment of papillomavirus warts. 2. The method of claim 1 wherein the purified lyophilized tuberculin is 1-50 TE. The process of claim 1, wherein the liquid carrier is sterile distilled water. 4. The process of claim 1 wherein the liquid carrier is physiological saline. 5. The method of claim 1 wherein the liquid carrier is a sterile 0.5 to 1% solution of lidocaine. 6. A process according to claim 1 wherein the additive is magnesium sulfate. 7. The process of claim 1, wherein the additive is calcium sulfate. 8. The process of claim 1 wherein the additive is a mixture of magnesium sulfate and calcium sulfate.