ES2437690B1 - Inhalation formulations in the form of solutions or dry powders, for the removal of mucous secretions from the respiratory system - Google Patents

Abstract

Inhalation formulations are described in the form of hypertonic solutions or dry powders free of preservatives or excipients containing an osmolarity agent chosen from NaCl or mannitol and hyaluronic acid of PM 300,000-350,000 obtained by fermentation of the bacterial strain Streptococcus equi, subspecies zooepidemicus, nonhemolytic

Description

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DESCRIPTION

Inhalation formulations in the form of solutions or dry powders, for the removal of mucous secretions from the respiratory system.

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The present invention relates to inhalation formulations in the form of hypertonic solutions or dry powders containing an osmolarity agent and hyaluronic acid of PM 300,000-350,000 obtained by fermentation of the bacterial strain Streptococcus equi, subspecies zooepidemicus, non-hemolytic or its salts .

10 State of the art

Mucociliary clearance is surely the main respiratory protection system. The mucus should thus have ideal characteristics to fully develop its important function of "entrapment" of foreign substances that enter the respiratory tract.

15 In different pathologies of the respiratory system the stagnation of the secretions in the respiratory tract and its consequent dehydration determines the establishment of conditions favorable to bacterial colonization and by
both to the worst evolution of the pathology in that place. The elimination of purulent mucus secretions represents a pharmacological objective of major importance.

20 In various genetic diseases, such as cystic fibrosis or Kartagener syndrome, this stagnation of dehydrated mucus in the respiratory tract determines a rapid deterioration of lung function, deterioration that can lead to early death. In other pathologies, such as in the BPCO, the stagnation of secretions and the subsequent bronchial obstruction occurs at a later age,

25 but in any case it can seriously compromise respiratory function.

Also in acute pathologies of viral origin, such as an insignificant flu, the stagnation of the mucous secretions is frequently responsible for the negative evolution of the respiratory picture.

30 Various mucolytic drugs, with mucolytic mechanism, have been used for years. Such drugs act by depolymerizing mucin, thanks to the presence of SH groups, thus reducing the viscosity of secretions.

Other molecules act as mucus kinetics by increasing the volume of fluid in the airways and thus inducing cough, thereby facilitating the separation of mucus from the airways.

35 Finally, active ingredients such as bromhexine and ambroxol have also been used for years, which stimulate the production of surfactant and thus facilitate the movement of mucus in the respiratory tract.

These mucoactive drugs have an even limited efficacy against cases of obstruction of moderate entity and

40 of acute evolution, but they are not very effective in diseases in which bronchial obstruction is more severe and especially chronic.

In cystic fibrosis, the malfunction of the CFTR channel protein, assigned to the ionic exchange of Na and chlorine between the cell and the extracellular environment, determines a reduction of the pericilliary fluid layer that covers the

45 respiratory mucosa. This surface liquid layer, kept constant with respect to the water entering and leaving the cell following the flow of the Na and Cl ions, is normally about 10 _M.

This superficial aqueous layer allows the cilia of the epithelium to move completely submerged in the periciliar fluid layer and thereby fully develop the task of eliminating secretions and all that there is

50 been caught by them for disposal. In subjects affected by CF, the pericilliary fluid layer is reduced by more than 50% to 4 _M, so that this reduction in volume reduces the ability of the cilia to move properly and therefore reduces, if it does not reach to cancel, mucociliary transport.

Because of the reduction in mucociliary clearance, there is a blockage of mucus in the pathways

55 as a result of dehydration, bacterial colonization and the development of ideal conditions for chronic colonization by pathogenic flora, especially Gram-.

The consequent activation of the immune system and the impossibility of eradicating the bacterial infection, lead to an inflammatory process in charge of the respiratory tract and to the progressive loss of respiratory function.

60 In other genetic pathologies, for example DCP, the causes that lead to the clearance deficit can be diverse, but the result is not different from that of CF, inflammation and chronic infection.

In fact, the evolution of CF, as well as that of DCP and chronic obstructive pathology (BPCO), is characterized by a marked inflammatory process that accompanies the infectious condition. In practice, in these

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Pathologies establish a vicious circle known as obstruction - infection - inflammation, a vicious cycle in which it is not yet clear what the exact sequence may be and in which the progressive self-feeding is clearly produced.

For many years now, systemic therapy has been accompanied by aerosol therapy for example with antibiotics such as tobramycin, colimycin, ciprofloxacin, aztreonam and with mucoactive agents such as, for example, dornase alfa, an enzyme capable of fragmenting the contained DNA. in mucus thus reducing viscosity.

The advantage of this route of administration is to reach the site of interest by reducing systemic participation, an important fact considering the chronicity of the treatment and the intrinsic toxicity of many of the drugs used.

All molecules used by inhalation have potentially risks for two reasons: they can favor the appearance of bronchospasm, and they can be irritating to the respiratory tract and therefore induce risks of undermining the scope of its pharmacological action, aggravating the pulmonary inflammatory condition. This risk is not only linked to the active substance only, but also to the formulation itself, to its physicochemical characteristics, as well as to the presence of possible excipients.

The formulation available by aerosols must be sterile to avoid the risk of transporting potential pathogens in the respiratory tract, must have a precise range of pH, osmolarity, and must give rise to particles of mean aerodynamic diameter less than 6 microns so that can reach the peripheral airways.

Among the most effective treatments to facilitate the secretion of the airways and used for a long time, solutions or dry powders of osmotic action have been used, such as saline hypertonic solutions

or mannitol The principle of operation of these treatments is linked to their ability to extract water in the respiratory tract, favoring the replacement of a superficial aqueous film with the consequent recovery of mucociliary clearance.

Treatment with solutions or powders of osmotic action is particularly effective particularly in pathologies such as CF and PCD, which are characterized by a marked bronchial obstruction due to a restoration of clearance. In these subjects, the high ionic concentration of NaCl or the high concentration of mannitol can exert osmotic pressure over time since it is not eliminated by mucociliary clearance, as in a normal subject, or as occurs in a subject affected by a lower mucociliary slowdown, for example chronic obstructive bronchitis.

Solutions of 3 to 9% NaCl for nebulization by means of a jet or mesh nebulizer, have long been used to hydrate the airways, improve ciliary clearance and therefore facilitate the elimination of mucopurulent secretions. More recently, dry powder formulations have been proposed to be inhaled, based on mannitol. The advantage of these dry powder formulations is that they reduce the time needed for inhalation and can potentially improve treatment compliance, especially in patients in whom the therapy should be prolonged.

The inhalation of hyperosmolar solutions of NaCl, as well as the inhalation of dry NaCl powder or the inhalation of dry mannitol powder, easily determine bronchial obstruction, throat irritation and cough also of considerable importance, effects that reduce the possibility If the airways are reached in depth, they reduce the respirable fraction of the product due to the alteration of the respiratory airflows and the consequent reduction in the effectiveness of the treatment.

It should also be noted that the taste of the hyperosmolar saline solution and the acceptability of inhalation of dry NaCl and mannitol powders are somewhat limited by the irritation and bronchoconstrictor effects induced by inhalation, thus more than 30% of Patients leave the treatment early.

In fact, a large number of patients do not risk regularly carrying out the treatment due to the unwanted effects that accompany the nebulization of saline hypertonic solutions or dry powders of osmotic agents.

The use of excipients and preservatives is highly advisable in view of the intended use and therefore the operating margins to improve the quality of the formulations seem somewhat limited. The excipients of normal use to improve the fluidity of the powders or to adjust the pH of the solutions or to increase the expiration date of the formulations in both solutions and powders, can develop an action favorable to inflammation on the respiratory epithelium and They are therefore inadvisable in aerosol formulations.

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Hyaluronic acid is a linear polysaccharide composed of glucuronic acid and N-acetylglucosamine belonging to the glycosaminoglycan family.

Hyaluronic acid is a natural component of the extracellular matrix and is present everywhere in many tissues. Hyaluronic acid has been used for years in medicine, in orthopedics, for intra-articular injection to increase the viscosity of synovial fluid, in ophthalmology, in otolaryngology and in cosmetic surgery.

Hyaluronic acid, as such, is not toxic and lacks allergenic effects.

For each of these activities different hyaluronic acids have been recommended either by their origin or by their molecular weight. More recently, high molecular weight hyaluronic acid has been suggested for respiratory use by virtue of its ability to reduce stress-induced bronchospasm (G. Petrigni, L. Allegra, Pulmonary Pharmacology & Therapeutics 19 (2006) 166-171 ).

In fact, another author (L.I.Z. Kunz et al., Pulmonary Pharmacology & Therapeutics 19 (2006) 286-291) showed that resorting to a lower PM hyaluronic acid had no activity in this regard.

The potential properties of hyaluronic acid, which must nevertheless still be confirmed, have been suggested by various authors with an objective such as pulmonary emphysema, chronic bronchitis and streptococcal infection.

DESCRIPTION OF THE INVENTION

The need for inhalation formulations with precise characteristics has so far limited the availability of products with ideal characteristics.

It has now been found that the addition of hyaluronic acid, or of its sodium salt, to formulations based on NaCl, in solution or dry powder, or of dry mannitol powder, improves the tolerability characteristics of inhalation by reducing induced bronchospasm by inhalation and, in the case of the solution, also the unpleasant salty taste that remains long after inhalation, causing nausea in the vast majority of patients.

It has been found in particular that it is possible to improve the tolerability of the inhalation of the osmotic substance by adding to it a possibly salified hyaluronic acid of PM 300,000-350,000 obtained by fermentation of the bacterial strain Streptococcus equi, subspecies zooepidemicus, nonhemolytic. Hyaluronic acid of extractive or animal or vegetable origin has proved surprisingly less effective, probably due to the presence of contaminating substances. In turn, it has been found that the specific hyaluronic acid allows a rapidly nebulizable solution to be obtained, characterized by particles with optimum mean aerodynamic diameter. Improved antibroncoconstrictive and anti-irritant efficacy is achieved. Such a finding contrasts with what has been reported in the literature that shows that hyaluronic acid is effective only at higher PM (approximately 1,000 KDa)

The exotoxin content of the formulations of the invention must be less than 0.1 ppm.

The hypertonic solution contains NaCl in a percentage between 6 and 12% w / v, preferably between 6 and 9% w / v and even more preferably 7% w / v. The biotechnological hyaluronic acid of PM comprised between

300,000 and 350,000 is between 0.05% and 0.3%, preferably between 0.05% and 0.2%, even more preferably equal to 0.1% w / v.

The solution to be inhaled is divided into small single dose bottles of 4-5 ml, and lacks excipients and buffers.

It has then been found that the solution of the invention, if distributed in single-dose vials of PELLD (low density polyethylene), is less susceptible to variations in pH with respect to polyethylene. In this way it was possible not to insert tampons.

The formulations of the invention in the form of hypertonic solutions contain NaCl and biotechnological hyaluronic acid of low molecular weight and lack excipients guaranteeing an optimum pH between 5 and 6.5, stable over time, and a longer duration of the formulation.

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It has also been found that, when inhaled as micronized dry powder formulations, osmolarity agents (NaCl and mannitol) are better tolerated, less bronchoconstrictors, and less toxigenic when associated with low molecular weight hyaluronic acid (LMW) in ratios. Osmolarity agent: acid

5 hyaluronic between 100: 0.05 and 100: 1.

The addition of hyaluronic acid described above also improves the creep of the powders by improving the pulmonary distribution.

10 Therefore, the object of the invention is formulations for inhalation in the form of hypertonic solutions or dry powders free of excipients such as preservatives, which contain an osmolar agent chosen from NaCl and mannitol and hyaluronic acid of PM 300,000-350,000 obtained by fermentation of the bacterial strain Streptococcus equi, subspecies zooepidemicus, non-hemolytic or its salts. The formulations in solution form are preferably distributed in single-dose containers of low density polyethylene while the

Powder formulations are preferably distributed in capsule.

Dust fogging can be carried out with any commercial device, both DPI and pMDI: DPI is preferred, that is devices activated by the patient's inspiratory act.

20 To evaluate the suitability of a nebulizer formulation, both the efficacy aspects and the possibility of carrying out the therapy in an easy, as well as fast, way should be taken into account, especially in those cases in which chronic use imposes considerable time effort on the patient.

The efficacy of different 7% NaCl solution formulations to which several hyaluronic acids are added has been evaluated in vitro, using Anderson Cascade Impactor technology through the assessment of

25 normalized parameters in the nebulizer, such as the average aerodynamic diameter, the particles being larger than 5 micrometers too thick to reach the secondary airways and easily impacting the larynx.

The respirable fraction indicates the percentage of nebulized active ingredient that effectively reaches the lower respiratory tract and is therefore available for therapeutic action.

30 The achievement of the desired action has been evaluated in vivo by the subjective assessment of the subject treated. The final criterion is expressed through the overall assessment of the suitability of the mist and its ability to perform the action in vivo.

Vitro: With the 4 formulations containing different hyaluronic acids, 10 healthy volunteers have been treated who have been required to express an opinion of 1-10 where 1 indicates a marked effect or negative opinion on the taste and 10 no effect or taste very Well, about the following aspects:

Fog time 5 ml using eflow rapid

Mean aerodynamic diameter Breathable dose% dose

7% NaCl AH at 500,000 - 700,000 Da

0.1% P.M 27 minutes > 7 twenty%

7% NaCl AH at 500,000 Da

0.1% P.M 23 minutes > 7 30%

7% NaCl at AH at 350,000 Da

0.1% P.M 10 minutes 4,5 75%

7% NaCl AH at 200,000 Da

0.1% P.M 10 minutes 5 75%

7% NaCl AH at

0.1% P.M 14 minutes 5 60%

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350,000 Da Animal origin

5 Protusigenic effect Bronchoconstriction Irritating effect on the mucosa Taste As a control solution, the nebulization of a hypertonic solution of 7% sodium chloride has been used.

10 The following table shows the average opinion of the 10 patients:

Cough

Bronchoconstriction Irritation Taste

7% NaCl

2 4 4 2

7% NaCl at 0.1% PM 500,000 Da

5 5 5 6

7% NaCl 0.1% AH 350,000 Da

8 8 10 9

7% NaCl 0.1% AH 200,000 Da

4 4 4 5

7% NaCl at 0.1% PM 350,000 Da Animal origin

7 2 4 7

As can be seen from the in vitro and in vivo data, the nebulized produced by the association of the 7% hypertonic solution with hyaluronic acid of fermentative origin that has an average PM of 15 350,000 has proved to be the best along with the PM 200,000 from the in vitro analysis, but without a doubt the best with respect to others in terms of the opinion of efficacy of the treated subjects.

The compositions of some formulations of the invention are set forth below by way of example.

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Examples

1. Small single-dose 5 ml low dose PE bottle

25 7% NaCl w / v

0.1% sodium hyaluronate (from Streptococcus equi, subspecies zooepidemicus, non-hemolytic, PM

300,000 - 350,000).

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Solution at pH 4.5-5.5.

2. Small 4 ml single dose bottle

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7% NaCl 0.1% Sodium Hyaluronate (from Streptococcus equi, subspecies zooepidemicus, nonhemolytic, PM

300,000 - 350,000).

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solution at pH 4.5 - 6.5

3. Small 5 ml single dose bottle

15 9% NaCl w / v 0.1% Sodium Hyaluronate (from Streptococcus equi, subspecies zooepidemicus, nonhemolytic, PM

300,000 - 350,000).

20 4. Small single dose bottle of 4 or 5 ml 6% NaCl w / v 0.1% sodium hyaluronate (from Streptococcus equi, subspecies zooepidemicus, nonhemolytic, PM

25 300,000 - 350,000).

pH of the solution 4-6.5.

5. Micronized powder inhalation capsule

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NaCl 350 mg

0.1% hyaluronic acid (from Streptococcus equi, subspecies zooepidemicus, non-hemolytic, PM 300,000 - 350,000).

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6. Micronized powder to inhale distributed in NaCl capsules, 350 mg

0.1% hyaluronic acid (from Streptococcus equi, subspecies zooepidemicus, non-hemolytic, PM 300,000 - 345,000) 0.35 mg.

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7. Micronized powder to inhale distributed in NaCl capsules, 250 mg

45 0.1% hyaluronic acid (from Streptococcus equi, subspecies zooepidemicus, non-hemolytic, PM 300,000 - 350,000) 0.25 mg.

8. Capsules to be inhaled from micronized powder, mannitol, 300 mg.

50 0.1% hyaluronic acid (from Streptococcus equi, subspecies zooepidemicus, non-hemolytic, PM 300,000 - 350,000)

9. Micronized powder to be inhaled distributed in 55 mannitol capsules, 420 mg

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0.1% hyaluronic acid (from Streptococcus equi zooepidemicus, non-hemolytic, PM 300,000 - 350,000)

Claims (7)

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Inhalation formulations in the form of hypertonic solutions or dry powders free of preservatives or excipients containing an osmolar agent chosen from NaCl or mannitol and 5 hyaluronic acid of PM 300,000-350,000 obtained by fermentation of the bacterial strain Streptococcus equi, subspecies zooepidemicus, nonhemolytic or one of its salts.
Inhalation formulations according to claim 1, in the form of hypertonic saline solutions in which sodium chloride is present in a percentage between 6 and 12% w / v, and the acid
Hyaluronic acid or one of its salts is present in a percentage between 0.05% and 0.3%. 3. Formulations according to claim 2, distributed in small single dose bottles of low density polyethylene. Formulations according to claim 2 or 3, wherein the osmolarity agent is present in percentages between 6 and 9%, preferably 7%.
5. Formulations according to claim 1, in the form of dry powders. twenty

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Formulations according to claim 5, wherein the ratio of osmolarity agent: hyaluronic acid is between 100: 0.05 and 100: 1.

7.

Formulations according to claim 5 or 6, wherein the osmolarity agent is sodium chloride.

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8.

Formulations according to claim 5 or 6, wherein the osmolarity agent is mannitol.

9.

Formulations according to any one of claims 1-8, for the removal of bronchial secretions.

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