WO2024063661A1

WO2024063661A1 - A pharmaceutical composition for topical application and use thereof in the treatment of hemangiomas in children

Info

Publication number: WO2024063661A1
Application number: PCT/PL2023/050078
Authority: WO
Inventors: Jerzy Niedzielski; Paweł Szymański; Kamila Czarnecka
Original assignee: Universytet Medyczny W Lodzi; Uniwersytet Medyczny w Lodzi
Current assignee: Universytet Medyczny W Lodzi; Uniwersytet Medyczny w Lodzi
Priority date: 2022-09-20
Filing date: 2023-09-20
Publication date: 2024-03-28
Anticipated expiration: 2025-03-20
Also published as: PL442316A1; EP4590288A1; PL244294B1

Abstract

The subject matter of the invention is a pharmaceutical composition for topical application containing a beta- adrenergic receptor antagonist and a corticosteroid and use thereof in the treatment of hemangiomas in children.

Description

A pharmaceutical composition for topical application and use thereof in the treatment of hemangiomas in children

Description

Technical Field

The subj ect matter of the invention is a pharmaceutical composition for topical application containing a beta- adrenergic receptor antagonist and a corticosteroid and use thereof in the treatment of hemangiomas in children .

Background Art

Hemangiomas are the most common mild tumors of early childhood that occur in children with an incidence of between 4 and 10 % . Early childhood angiomas develop on the face and neck ( approx . 80 % of cases ) as well as on the trunk and limbs . As early as in the first weeks of li fe , a phase of rapid proli ferative growth of angiomas starts which extends until the end of the child' s first year of age . The subsequent phase of slow regression di f fers in its duration and may extend until the age of 5-10 years . Approx . 10-15 % of early childhood angiomas grow to si zes that cause functional impairment of an organ or are li fe-threatening to the child . Systemic therapy and/or surgical treatment need to be started immediately in such patients . Several therapeutic modalities are available for the other cases of angiomas : laser therapy, scleroti zation, cryotherapy, surgical excision, which require the patient to undergo general anesthesia . Another approach is the oral administration of cardiac medicines (propranolol ) ( 2 ) which accelerate the regression of vascular lesions , frequently all the way to their complete disappearance . Their ef fect on angiomas was revealed by accident ; nevertheless , they are currently routinely used worldwide based on accepted standards owing to their high efficacy. An inconvenient aspect of the therapy is the need for initial hospitalization (2-3 days) , an ECG to be performed and a cardiology consultation to evaluate the patient's eligibility for therapy .

To eliminate these disadvantages, topical therapy has started to be used through the superficial application of medicines in the form of cream, gel or ointment on angiomas (1, 3, 4) . These are formulated on demand based on commercially available ready-made products, such as ophthalmic drops (timolol) (4) , oral tablets or intravenous injections (propranolol) (3) , that is, medicines that according to manufacturers' instructions should be applied via different routes. In addition, intralesional therapies are known, which involve an injection of a formulation inside a lesion; the therapy, however, requires general anesthesia and, therefore, hospitalization and involvement of specialized staff, with a potential risk of local and systemic complications.

The efficacy of any medicines administered was not tested in any of the therapeutic modalities previously used in practice, including those listed above, and their therapeutic doses considering the different application method or synergy of action, in particular combinations with corticosteroids, were not determined.

A publication of Peng Xu et al. : "A self-controlled study of intralesional injection of diprospan combined with topical timolol cream for treatment of thick superficial infantile hemangiomas"; Dermatologic Therapy 2018, 31, el2595; DOI: 10 .1111/dth .12595 disclosed a combination therapy of hemangiomas in children which involved an intralesional injection of diprospan and topical application of cream containing 0.5 % timolol.

A publication of Alahmady H. Alsmman et al. : "Combined oral propranolol with intralesional injection of triamcinolone acetonide in treatment of infantile periocular hemangiomas"; Clinical Ophthalmology 2017, 11, 2177-2181; http://dx.doi.org/10.2147/OPTH.S153121 disclosed a combination therapy of hemangiomas in children which involved oral administration of propranolol and intralesional injection of triamcinolone acetonide.

A publication of Wenhu Zhou et al. : "Formulation, characterization and clinical evaluation of propranolol hydrochloride gel for transdermal treatment of superficial infantile hemangioma"; Drug Development and Industrial Pharmacy 2014, Informa Healthcare USA, Inc.; DOI: 10.3109/03639045.2014.931968 disclosed a manufacturing process for a pharmaceutical composition in the form of gel, containing propranolol hydrochloride, HPMC, a terpene, alcohol and other excipients for treating hemangiomas in children .

A publication of Retno Danarti et al. Topical Timolol Maleate 0.5 % for Infantile Hemangioma: Its Effectiveness Compared to Ultrapotent Topical Corticosteroids - A SingleCenter Experience of 278 Cases. Dermatology 2016; 232: 566- 571 disclosed a comparison of the efficacy of steroid ointment with ointment containing timolol.

Summary of Invention

The objective of the invention was to develop a pharmaceutical composition for topical application effective in the treatment of hemangiomas in children. The subj ect matter of the invention is a pharmaceutical composition for topical application, characteri zed in that it contains a beta-adrenergic receptor antagonist , a corticosteroid and a pharmaceutically acceptable carrier, wherein the beta-adrenergic receptor antagonist is labetalol or a pharmaceutically acceptable salt thereof , and the corticosteroid is selected from a group including triamcinolone , triamcinolone acetonide , triamcinolone hexacetonide and triamcinolone diacetate .

Preferably, the pharmaceutically acceptable salt of labetalol is the hydrochloride .

Preferably, the weight ratio of the beta-adrenergic receptor antagonist to the corticosteroid is 1 : 3 .

Preferably, the concentration of the beta-adrenergic receptor antagonist in the composition is 0 . 02-0 . 5 % by weight .

Preferably, the concentration of the corticosteroid in the composition is 0 . 02-0 . 5 % by weight .

Preferably, the composition is in the form of cream, ointment or gel .

Another subj ect matter of the invention is a pharmaceutical composition as defined before for use in the treatment of hemangiomas in children .

An advantage of the invention is that one formulation is used instead of several formulations . Two substances from two distinct therapeutic groups ( a beta-adrenergic receptor antagonist and a corticosteroid) , previous ly used separately, are combined in one formulation . Therefore , concomitant use of several formulations , whose combined ef fects ( antagonism, synergy) are unknown, is avoided . This is not only convenient but first of all safe , both for the child (patient) and for the person who applies the formulations (errors may occur when several formulations are used; the dose as required is ensured) .

Another advantage of the invention is that the formulation is used externally. Even though most angiomas are located within the skin, previous therapeutic methods were based on the administration of formulations by mouth or in the form of injections, thus having systemic effects on the body; this involves a need for general anesthesia, hospitalization and hiring specialized personnel in small children. The pharmaceutical composition of the invention is used topically, that is, only in the affected site, which definitely reduces the systemic effects of medicines administered. Therefore, the risk of undesirable effects due to the medicines administered, and thus their adverse effect on the child's health, is minimized. The solution as developed facilitates the application of the formulation at home by the family. Hospitalization or involvement of medical professionals is not required, which is convenient for parents and children.

Another advantage of the invention is a reduced dosage of medicines administered. The efficacy of any therapy depends on the administration of required drug doses. For example, beta-adrenergic receptor antagonists (betablockers) administered by mouth have systemic effects, first of all on the cardiovascular system when used as indicated. After achieving the required therapeutic level in the patient's blood, they act by saturating all tissues and organs with the therapeutic substance. Vascular lesions in the skin are not the principal therapeutic target of betablockers, and a side effect is rather involved. Owing to the pharmaceutical composition of the invention, drug doses administered topically ( externally) can be reduced, because there is no need for loading the patient ' s whole body to achieve a drug concentration so that it can penetrate the lesion ( from inside ) . When administered topically, the pharmaceutical composition of the invention acts directly on the lesion . In addition, based on the results of in vitro studies into the synergy of medicines combined in appropriate concentrations ( the corticosteroid and the beta-blocker ) , the doses could be reduced to increase the strength of action compared to conventionally used concentrations of the drugs administered separately : additive synergy was achieved .

Examples

Example 1

Determination of IC50 for selected compounds

In the experiment , an angioma cell line was used : EOMA (ATCC® CRL-2586™) Hemangioendothel ioma . The cells were grown in the EMEM (ATCC ) medium and seeded in a 96-well plate for 24 hours . Subsequently, they were incubated with the test compound in selected concentrations : 50-350 pM ( the compound was dissolved in DMSO and water ) for 48 hours . The final methanol concentration in each well was 1 pL per 100 pL of the medium . The EMEM medium with 1 pL methanol added was used as control and pure methanol was the positive control . A cytotoxicity test was performed after 48 h of incubation according to the Vybrant MTT cell proli feration test kit : 10 pL of 12 mM MTT stock solution was added to the cells and further incubated at 37 ° C for 4 h . Volume of 25 pL was removed from the wells after incubation and 50 pL DMSO was added . The plates were incubated at 37 ° C for 10 min and absorbance at 540 nm was read . The test compounds were four compounds acting on beta-adrenergic receptors : propranolol hydrochloride , labetalol hydrochloride , atenolol and isoprenaline hydrochloride, and four corticosteroids: prednisolone, prednisone, hydrocortisone, and triamcinolone.

IC50 values for all eight test compounds were determined as a result of tests in cell lines. The lowest IC50 values were found for labetalol hydrochloride and triamcinolone. These were 308 pM and 844 pM, respectively.

Example 2

In vitro evaluation of the labetalol hydrochloride and triamcinolone combination in various concentrations

Within stage two of the studies, a model was developed to test the therapeutic efficacy and interactions between the effects of labetalol hydrochloride and triamcinolone in various concentrations. The study was conducted using angioma cell lines: EOMA (ATCC® CRL-2586™) Hemangioendothelioma .

A stock solution (S2) of triamcinolone with a concentration of 1 688.5 pM was prepared. Subsequently, a stock solution (SI) of labetalol hydrochloride with a concentration of 617.8 pM was prepared. The assumption was to prepare a combination of concentrations within the IC50 values determined in cell lines for the test compounds in stage one of the studies. Table 1 below shows how combinations of concentrations were prepared to determine the best synergic action of both substances.

Table 1

51 - stock solution of labetalol hydrochloride = 617 . 8 piM

52 - stock solution of triamcinolone = 1 688 . 5 piM

Sixteen combinations of various concentrations of the test substances were obtained as shown in the diagram below .

Subsequently, the resulting 16 combinations were tested in cell lines . The average viability for all combinations was determined . In the experiment , an angioma cell line was used : EOMA (ATCC® CRL-2586™) Hemangioendothel ioma . The cells were grown in the EMEM (ATCC ) medium and seeded in a 96-well plate for 24 hours . Subsequently, they were incubated with the test compound in selected concentrations as listed in Table 1 for 48 hours . The final methanol concentration in each well was 1 pL per 100 pL of the medium . The EMEM medium with 1 pL methanol added was used as control and pure methanol was the positive control . A cytotoxicity test was performed after 48 h of incubation according to the Vybrant MTT cell proli feration test kit : 10 pL of 12 mM MTT stock solution was added to the cells and further incubated at 37 ° C for 4 h . Volume of 25 pL was removed from the wells after incubation and 50 pL DMSO was added . The plates were incubated at 37 ° C for 10 min and absorbance at 540 nm was read .

Conclusions

When the concentrations calculated based on IC50 values for the compounds were used, synergy was achieved; therefore , the ef fect of two substances used in speci fic concentrations resulted in the enhanced activity of the mixture of triamcinolone and labetalol compared to the ef fect of the drugs administered separately. It was found that labetalol hydrochloride used in the IC50 concentration in combination with small amounts of triamcinolone had an effect on the reduced viability of angioma cells. Therefore, it was shown that synergy occurred already at low corticosteroid concentrations; as a result, the dose thereof in combination with the beta-blocker could be reduced while achieving the same therapeutic effect. The use of a mixture of both test substances achieved better efficacy against angioma cancer cells than when each substance was used separately. Therefore, an effect of additive synergy was achieved.

The results of studies in angioma cells in the concentration range tested for the combination of test compounds showed that the test compounds had synergy of action for all combinations of concentrations. However, the highest synergy of action was found at labetalol hydrochloride and triamcinolone concentrations as determined for mixture no. 13.

Example 3

Preparation of pharmaceutical compositions

(a) Preparation of cream

0.5 g triamcinolone and 0.166 g labetalol hydrochloride, respectively, were weighed out, placed in a mortar and micronized. Subsequently, the weighed cream base was added and ground until a homogenous mass was obtained.

If the recipe lists a solution, micronized substances are dissolved in the solution. The weighed cream bases were ground in a mortar, and a solution containing labetalol hydrochloride and triamcinolone was gradually added. The grinding was continued until a homogenous mass was obtained. Recipe: 1 : 3 mixture (0.166 g labetalol hydrochloride ;

0.5 g triamcinolone )

3 % boric acid solution 15.0

Lekobaza to 100.0

Cream was prepared .

Recipe: 1 : 3 mixture (0.166 g labetalol hydrochloride ;

0.5 g triamcinolone )

Distilled water

Hascobaza in equal parts to 100.0

Cream was prepared .

Recipe: 1 : 3 mixture (0.166 g labetalol hydrochloride ;

0.5 g triamcinolone )

Distilled water q . s .

Lanolin 20.0

Petroleum jelly to 100.0

Cream was prepared .

Recipe: 1 : 3 mixture (0.166 g labetalol hydrochloride ;

0.5 g triamcinolone )

Eucerin

Distilled water in equal parts to 100.0

(b) Preparation of ointment

0.5 g triamcinolone and 0.166 g labetalol hydrochloride, respectively, were weighed out, placed in a mortar and micronized. Subsequently, the weighed ointment base was added and ground in the mortar until a homogenous mass was obtained.

1) Recipe: 1 : 3 mixture (0.166 g labetalol hydrochloride;

0.5 g triamcinolone)

Cholesterol ointment to 100.0

Ointment was prepared.

2) Recipe: 1 : 3 mixture (0.166 g labetalol hydrochloride;

0.5 g triamcinolone) Petroleum jelly to 100.0 Ointment was prepared.

3) Recipe: 1 : 3 mixture (0.166 g labetalol hydrochloride;

0.5 g triamcinolone) Liquid paraffin 10.0

Yellow petroleum jelly to 100.0 Ointment was prepared.

4) Recipe: 1 : 3 mixture (0.166 g labetalol hydrochloride;

0.5 g triamcinolone) Resorcinol 5.0

Liquid paraffin 5.0 Ichthyol 2.0 Zinc paste to 100.0

Ointment was prepared.

(c) Preparation of gel

0.5 g triamcinolone and 0.166 g labetalol hydrochloride, respectively, were weighed out, placed in a mortar and micronized. Subsequently, the weighed gel base was added and ground until a homogenous mass was obtained.

If the recipe lists a solution, micronized substances are dissolved in the solution. The weighed gel bases were ground in a mortar, and a solution containing labetalol hydrochloride and triamcinolone was gradually added. The grinding was continued until a homogenous mass was obtained.

1) Recipe: 1 : 3 mixture (0.166 g labetalol hydrochloride;

0.5 g triamcinolone)

2 % lignocaine gel to 50.0

Gel was prepared.

2) Recipe: 1 : 3 mixture (0.166 g labetalol hydrochloride;

0.5 g triamcinolone) Methylcellulose ointment to 100.0 Gel was prepared. 3) Recipe: 1 : 3 mixture (0.166 g labetalol hydrochloride;

0.5 g triamcinolone)

Menthol 2.0

Glycerol ointment to 50.0

Gel was prepared.

4) Recipe: 1 : 3 mixture (0.166 g labetalol hydrochloride;

0.5 g triamcinolone)

Menthol 5.0

Capsicum tincture 2.0

2 % lignocaine gel Macrogol ointment in equal parts to 100.0

Gel was prepared.

Reference list

(1) Peng Xu et al. : "A self-controlled study of intralesional injection of diprospan combined with topical timolol cream for treatment of thick superficial infantile hemangiomas"; Dermatologic Therapy 2018, 31, el2595; DOI: 10.1111/dth.12595;

(2) Alahmady H Alsmman et al. : "Combined oral propranolol with intralesional injection of triamcinolone acetonide in treatment of infantile periocular hemangiomas"; Clinical Ophthalmology 2017, 11, 2177- 2181; http://dx.doi.org/10.2147/OPTH.S153121;

(3) Wenhu Zhou et al. : "Formulation, characterization and clinical evaluation of propranolol hydrochloride gel for transdermal treatment of superficial infantile hemangioma"; Drug Development and Industrial Pharmacy 2014, Informa Healthcare USA, Inc.; DOI: 10.3109/03639045.2014.931968;

(4) Retno Danarti et al. : "Topical Timolol Maleate 0.5 % for Infantile Hemangioma: Its Effectiveness Compared to Ultrapotent Topical Corticosteroids - A SingleCenter Experience of 278 Cases"; Dermatology 2016, 232, 566-571.

Claims

Claims A pharmaceutical composition for topical application, characterized in that said composition contains a beta- adrenergic receptor antagonist, a corticosteroid and a pharmaceutically acceptable carrier, wherein the beta- adrenergic receptor antagonist is labetalol or a pharmaceutically acceptable salt thereof, and the corticosteroid is selected from a group including triamcinolone, triamcinolone acetonide, triamcinolone hexacetonide and triamcinolone diacetate. The composition of Claim 1, characterized in that the pharmaceutically acceptable salt of labetalol is the hydrochloride . The composition of Claim 1 or Claim 2, characterized in that the weight ratio of the beta-adrenergic receptor antagonist to the corticosteroid is 1 : 3. The composition of any one of Claims 1 to 3, characterized in that the concentration of the beta- adrenergic receptor antagonist in the composition is from 0.02 to 0.5 % by weight. The composition of any one of the preceding Claims 1 to

4, characterized in that the concentration of the corticosteroid in the composition is from 0.02 to 0.5 % by weight. The composition of any one of the preceding Claims 1 to

5, characterized in that said composition is in the form of cream, ointment or gel. A pharmaceutical composition as defined in Claims 1 to 6 for use in the treatment of hemangiomas in children.