KR20010014163A - Proliposome Powders for Inhalation Stabilised by Tocopherol - Google Patents

Abstract

The present invention provides a proliposomal powder comprising separated particles comprising (1) a biologically active ingredient, (2) a stabilizing portion of tocopherol, and (3) a lipid or lipid mixture having a phase transition temperature of 37 ° C. or less in a single phase. do. Also disclosed are methods of preparing proliposomal powders and pharmaceutical uses.

Description

Proliposome Powders for Inhalation Stabilised by Tocopherol

Liposomes are membrane like vesicles consisting of a series of concentric lipid bilayers that alternate with hydrophilic compartments. Liposomes can be made from a variety of natural and synthetic lipids, such as natural and synthetic phosphoglycerol lipids, spinolipids and digalactosylglycerol lipids. One of the main uses of liposomes has been the use of different kinds of pharmaceutical active ingredients as carriers to improve drug delivery and minimize the side effects of some treatments. The pharmaceutical active ingredient may be encapsulated in the hydrophilic compartment of the liposome (if the active ingredient is water soluble) or encapsulated into the lipid bilayer if the active ingredient is lipophilic and incorporated into the liposome.

One of the major problems associated with liposome pharmaceutical formulations is their long term stability. Aqueous liposome dispersions have limited stability due to aggregation, loss of the encapsulated active ingredient to the external phase, chemical degradation of the active ingredient or lipid material, and the like.

These problems can be mostly overcome if a solid composition is used. Such solid compositions may comprise liposome powders, ie dry liposome dispersions or proliposomal powders.

International publication WO96 / 19199 refers to various liposome and proliposomal documents and describes proliposomal powders. The powder contains discrete particles containing biologically active ingredients and lipids or lipid mixtures with a phase transition temperature (Tc) of 37 ° C. or less in a single phase.

It has finally been found that the stability of the proliposomal powder of International Publication No. WO96 / 19199 can be increased to a considerable extent.

The present invention relates in particular to inhaled proliposomal powders, to methods for producing proliposomal powders, to proliposomal powder-containing compositions and methods of use thereof.

The present invention provides a proliposomal powder comprising separated particles comprising (1) a biologically active ingredient, (2) a stabilizing portion of tocopherol, and (3) a lipid or lipid mixture having a phase transition temperature of 37 ° C. or less in a single phase. do.

Preferably the tocopherol is α-tocopherol, more preferably racemate α-tocopherol.

The powder is particularly suitable for administration by inhalation.

Single phase powders may alternatively be described as comprising biologically active ingredients, lipids or mixtures of lipids with a phase transition temperature of 37 ° C. or less, and homogeneous molecular mixtures of tocopherols.

It will be appreciated from the terms "single phase" and "uniform molecular mixture" that there is no separate crystalline phase of the active ingredient, lipid or tocopherol in the powders of the invention.

Single phase powders may be inhaled directly and simultaneously, for example into the upper or lower portion of the respiratory tract, and will form liposomes incorporating biologically active ingredients.

In general, any amphiphilic lipid or lipid mixture known to be suitable for preparing liposomes by known methods can be used in the present invention. The lipid or lipid mixture must have a phase transition temperature below body temperature (37 ° C.) for proliposomal powder products that can be hydrated under physiological conditions (ie to be able to form liposomes in the respiratory tract). Phase transition temperatures of different lipid mixtures are readily measured using satisfactorily established methods, for example DSC (see, eg, J. Suurkuusk et al., Biochemistry, vol. 15, no. 7, p. 1393 (1976)). can do. In general, any natural or synthetic lipid or lipid mixture having a phase transition temperature of 37 ° C. or lower is usable in the present invention.

Examples of potentially useful lipids may include natural and synthetic lipids, such as natural and synthetic phosphoglycerol lipids, spinolipids and digalactosylglycerol lipids. Spinolipids (SL), such as spingomierin (SM), ceramide and cerebromide, among natural lipids; Galactosylglycerol lipids such as digalactosyldiacylglycerol (DGalDG); Phosphoglycerol lipids such as egg yolk phosphatidylcholine (e-PC) and soybean phosphatidylcholine (s-PC); Mention may be made of lecithin, such as egg yolk lecithin (e-lecithin) and soybean lecithin (s-lecithin). Among the synthetic lipids are dimyristoyl phosphatidylcholine (DMPC), dipalmitoyl phosphatidylcholine (DPPC), distearoyl phosphatidylcholine (DSPC), dilauryl phosphatidylcholine (DLPC), 1-myristoyl-2-palmitoyl phosphatidylcholine (MPPC) ), 1-palmitoyl-2-myristoyl phosphatidylcholine (PMPC), and dioleoyl phosphatidylcholine (DOPC). Among lipid mixtures, SM / PC, SM / cholesterol, ePC / cholesterol, sPC / cholesterol, PC / PS / cholesterol, DMPC / DPPC, DMPC / DPPC / CH, DMPC / CH, DPPC / DOPC, DPPC / DOPC / CH, DLPC / DPPC, DLPC / DPPC / CH, DLPC / DMPC, DLPC / DMPC / CH, DOPC / DSPC, DPSM / DMSM, e-lecithin / cholesterol and s-lecithin / cholesterol may be mentioned. In addition to dimyristoyl phosphatidylglycerol (DMPG), diphosphalmitoyl phosphatidylglycerol (DPPG), dimyristoyl phosphatidic acid (DMPA), dipalmitoyl phosphatidic acid (DPPA) or stearylamine (SA) Charged lipids.

Lipids of particular interest in the present invention are DPPC and / or DMPC. Preference is given to mixtures of DPPC and DMPC comprising at least 10% by weight DMPC (eg 10-50% DMPC). Particularly preferred are mixtures of DPPC and DMPC containing one or more charged lipids, for example DMPG, DPPG, DMPA or SA in an amount up to 5% by weight. Another preferred mixture includes DPSM and DMSM, optionally containing one or more charged lipids, and a mixture of e-lecithin or s-lecithin and cholesterol, optionally containing one or more charged lipids, with Tc less than 37 ° C. Other mixtures can be readily selected by those skilled in the art, for example with reference to Gregor Cevc, Phospholipids Handbook, Marcel Dekker, New York (1993) pp 427-435.

Tocopherol is present in a proportion of preferably 0.05 to 1.0% by weight, more preferably 0.1 to 0.6% by weight of the total single phase containing the lipid (s) and the biologically active component.

The active ingredient preferably has a molecular structure that can be incorporated into the lipid bilayer to assist in encapsulation in liposomes upon hydration. Examples of such molecular structures are fatty acid esters with long chain hydrocarbons sufficient to act as hydrophobic groups.

Suitable active ingredients can be readily recognized by those skilled in the art and include anti-inflammatory and bronchiolytic agents and antihistamines, cyclooxygenase inhibitors, leukotriene synthesis inhibitors, leukotriene antagonists, phospholipase-A2 (PLA2) inhibitors, platelet aggregation factor (PAF) ) Antagonists, asthma preventive agents, and the like. Antiarrhythmic medications, mental stabilizers, cardiac glucosides, hormones, antihypertensive medications, antidiabetics, antiparasitic and anticancer agents, sedatives, analgesics, antibiotics, antirheumatics, inflammatory drugs, antifungals, antihypertensives, vaccines, antiviral agents, proteins Peptides and vitamins may also be of interest.

Specifically, budesonide, fluticasone propionate, ciclesonide, for example loplefonide as its palmitate, for example mometasone, tripredan, RPR 106541, dexamethasone, Glucocorticosteroids and derivatives thereof such as betamethasone, fluorinolone, flumethasone, triamcinolone acetonide, gluronisonide, beclomethasone and 16, 17-acetal of pregnane derivatives; And β-2 agonists such as terbutalin, salmeterol, salbutamol, formoterol, phenoterol, clenbuterol, procaterol, bitolterol, and broxaterol, can be used in the present invention. The active ingredient may also be a mixture of pharmaceutically active ingredients, for example a mixture of glucocortico-steroids and bronchodilators such as formoterol, salmeterol, terbutalin or salbutamol may be used. For the avoidance of doubt when referred to herein for any active ingredient, the above reference is intended to include references to pharmaceutically acceptable esters, salts and hydrates thereof.

When the active ingredient is a steroid, steroid esters are preferred.

The active ingredient is preferably a steroid, preferably a steroid esterified at the 21-position with saturated or unsaturated fatty acids of at least 8, for example at least 10 or at least 12 carbon atoms. Fatty acids may have, for example, up to 24 carbon atoms, for example up to 20 carbon atoms or up to 18 carbon atoms. More preferably, the active ingredient is steroid-21-palmitate, myristate, laurate, caprate, caprylate or stearate. The most preferred active ingredient according to the invention is (22R) -16α, 17α-butylidenedioxy-6α, 9α-difluoro-11β-hydroxy-21-palmitoyloxypregin-4-ene-3,20 -Dione, lofloponide palmitate compound.

When the active ingredient is an ester it must be able to hydrolyze into the main active material. Surprisingly, the single phase proliposomal powders of the present invention promote the required hydrolysis in the reaction system, whereas the crystalline esters will generally not hydrolyze.

When delivery by inhalation is desired, as much of the proliposomal powder of the invention as possible is particles less than 10 microns in diameter, for example 0.01-10 microns or 0.1-6 microns, for example 0.1-5 microns, or It must consist of agglomerates of particles. Preferably at least 50% of the powder forms particles in the preferred size range. For example, at least 60%, preferably at least 70%, more preferably at least 80% and most preferably at least 90% of the powders form particles or agglomerates of these particles within the desired size range.

The proliposomal powder of the present invention does not need to contain other ingredients. However, pharmaceutical compositions containing the powders of the present invention may also include other pharmaceutically acceptable additives such as pharmaceutically acceptable adjuvants, diluents and carriers. These may be added to the proliposomal composition after any micronization or before any micronization if the solvent is completely removed. Any carrier is preferably a crystalline hydrophilic material. Preferred carriers are crystalline lactose monohydrate. Other suitable carriers include glucose, fructose, galactose, trehalose, sucrose, maltose, raffinose, maltitol, melezitose, stachiose, lactitol, pallatinite, starch, xylitol, mannitol, myoinositol and the like and the like Hydrates and amino acids such as alanine and betaine.

The amount of additive present in the formulation can vary over a wide variety of ranges. In some cases, little or no additive may be required, while it is often desirable to dilute the powder with additives, for example, to improve the powder properties used in the inhaler. In the latter case, at least 50%, for example at least 70% or at least 80% of the formulation may consist of additives, with the remainder being proliposomal powders. The percentage of additives may also depend on the efficacy of the biologically active compound and the optimum amount of powder for inhalation.

When an additive, for example a carrier, is present, the total composition may be in the form of particles of a size within the respirable particle size range. Alternatively, the carrier may comprise coarser particles of mass median particle size greater than 20 microns, for example, or comprise agglomerates of smaller particles, so that in either case a prescribed mixture of proliposomes and carriers can be formed The mass may, for example, have a mass median particle diameter of greater than 20 microns.

A further object of the present invention relates to the provision of a process for the preparation of the proliposomal powder of the invention, i.

Thus, the present invention also relates to a crystalline solvent matrix in the free state by dissolving lipids or mixtures of lipids, tocopherols and lipophilic biologically active components in solvents and freezing below the phase transition temperature on lipids, with phase transition temperatures up to 37 ° C. And obtaining a single lipid phase and evaporating the frozen solvent below the phase transition temperature of the lipid phase.

Freezing of the solution and solvent evaporation can be carried out by conventional methods, for example in a conventional freeze-dryer. For example, a solution of lipids, tocopherols and biologically active ingredients is poured onto a shelf in a freeze dryer and frozen by lowering the temperature. Solvent evaporation can then be achieved, for example, by lowering the pressure of the freeze drying chamber, and the resulting powder can be stripped from the shelf of the chamber and optionally passed through a sieve.

The freeze-dried powder may be further processed if necessary to obtain particles within the respirable particle size range, for example, the freeze-dried powder may be micronized using, for example, an air jet mill to provide respirable particles. Can be.

Freezing of the solution of the biologically active component, tocopherol and lipids is carried out by the method of producing a single lipid phase in the frozen solvent matrix. The production of a single lipid phase is controlled by the final temperature and the rate of freezing of the solution, and the optimum rate of freezing of any particle solution may be between the time needed for crystallization of the solvent in question and the time required for crystallization of lipids, tocopherols and active ingredients. This will simply be determined by one skilled in the art through trial and error. The optimal final temperature should be 10-20 ° C. below the glass transition temperature of the lipid phase. For example, powder X-ray methods can be used to monitor crystallinity and differential scanning calorimetry can be used to monitor the extent of incorporation of biologically active ingredients into liposomes after hydration.

Since it is essential that all ingredients are in solution prior to freezing to avoid precipitation or phase separation which causes the powder to have more than one phase, the solvent must have the ability to dissolve the lipids, tocopherols and biologically active ingredients completely. In addition, the solvent should be toxicologically acceptable and have a suitable freezing point and preferably high vapor pressure. The solvent can be, for example, an organic solvent, for example an alcohol, or a mixture of aqueous and organic solvents. Preferred solvents for use in the present invention are tert-butanol.

The powder can optionally be agglomerated into small spheres to control the cohesiveness of the powder. The spheres should preferably not be larger than 1 mm in diameter, and spheres larger than this can be removed, for example, by sieving. Any agglomerates are brittle and they can easily deaggregate, for example, in the air stream from the powder inhaler.

Proliposomal powders of the invention are useful for local or systemic treatment of a disease and may be administered through the upper and lower portions of the respiratory tract, including, for example, the route of the nose. As such, the present invention also provides the above proliposomal powder for therapeutic use, the use of the proliposomal powder in the manufacture of a medicament for the treatment of diseases through the respiratory tract, and a therapeutically effective amount of the proliposomal powder of the present invention to the patient. Provided are methods of treating a patient in need thereof.

For example, the proliposomal powder of the present invention can be used for the treatment of inflammatory diseases of the respiratory tract, such as asthma, rhinitis, alveolitis, bronchitis, and bronchitis.

Administration to the respiratory tract may be carried out using, for example, a dry powder inhaler or a pressurized aerosol inhaler.

Suitable dry powder inhalers are intended for use in multiple doses, such as, for example, single dose inhalers known under the trademark Monohaler® and for example inhalers known under the trademark Turbuhaler®. Breath-acting dry powder inhaler.

While the proliposomal powders of the present invention are particularly applicable for administration by inhalation, they may also be included in the formulations applied for other forms of delivery. For example, oral, topical and injectable preparations can be prepared, for example, for the treatment of inflammatory joint diseases such as arthritis, skin diseases and visceral diseases.

The following examples are intended to illustrate but not limit the scope of the invention. Parts are parts by weight.

<Example 1>

Loplefonide palmitate (10 parts), DPPC (63 parts), DMPC (24 parts), NaDPPG (3 parts) and racemate α-tocopherol (0.1 parts) at 80 ° C. in tert-butanol (1300 parts) Dissolved. The solution was poured into a rack of freeze dryer cooled to -35 ° C. The solution reached this temperature after about 30 minutes and then reduced the pressure in the freeze dryer to induce sublimation of the solvent. The sublimation rate can be controlled by decreasing the pressure and increasing the temperature, while the process temperature is kept no higher than -10 ° C. Freeze drying was continued until all solvent was removed. The resulting powder was stripped off the shelf of the freeze dryer and passed through the sieve.

The powder was micronized in an air jet mill so that the powder particle size was less than 5 μm. The micronized powder was mixed with lactose monohydrate (20 parts powder: 80 parts lactose monohydrate) by a sieving process, and the mixture was further homogenized by micronization in a jet mill at low pressure.

The powder mixture was agglomerated into spheres no greater than 1 mm using standard techniques. Larger spheres were removed by sieving. The agglomerated powder was filled into a Turbuhaler® dry powder inhaler.

In a separate experiment the amount of α-tocopherol in the formulation was changed to 0.06 parts and 0.6 parts, respectively.

It was surprisingly found that the proliposomal formulation of Example 1 was more stable than the corresponding formulation containing other antioxidants.

Powder Analysis

X-ray powder diffraction was performed on the powder mixture of Example 1 to find that no crystalline state was present in the powder.

Incorporation of active ingredients into liposomes

The proliposomal powders of Examples 1 and 2 were hydrated and the degree of incorporation of the active ingredient was measured using differential scanning calorimetry. DSC showed that the active ingredient was incorporated sufficiently into the liposome.

Claims (27)

A proliposomal powder comprising separated particles comprising (1) a biologically active ingredient, (2) a stabilizing portion of tocopherol, and (3) a lipid or lipid mixture having a phase transition temperature of 37 ° C. or less in a single phase. The proliposomal powder of claim 1, wherein the tocopherol is α-tocopherol. The proliposomal powder of claim 2, wherein the tocopherol is racemate α-tocopherol. The proliposomal powder according to any one of claims 1 to 3, wherein the tocopherol is present in a proportion of 0.05 to 1.0% by weight of the single phase. The proliposomal powder of claim 4, wherein the tocopherol is present at a ratio of 0.1 to 0.6% by weight. The proliposomal powder according to any one of claims 1 to 5, comprising at least one lipid selected from the group consisting of natural and synthetic phosphoglycerol lipids, spinolipids and digalactosylglycerol lipids. The method according to any one of claims 1 to 6, wherein spingomierine (SM) / phosphatidylcholine (PC), SM / cholesterol, egg yolk phosphatidylcholine (ePC) / cholesterol, soybean phosphatidylcholine (sPC) / cholesterol, PC / PS / Cholesterol, dimyristoyl phosphatidylcholine (DMPC) / dipalmitoyl phosphatidylcholine (DPPC), DMPC / DPPC / CH, DMPC / CH, DPPC / dioleoyl phosphatidylcholine (DOPC), DPPC / DOPC / CH, dilauryl phosphatidylcholine (DLPC ) / DPPC, DLPC / DPPC / CH, DLPC / DMPC, DLPC / DMPC / CH, DOPC / Distearoyl phosphatidylcholine (DSPC), DPSM / DMSM, e-lecithin / cholesterol and s-lecithin / cholesterol Proliposomal powder comprising the selected lipid mixture. 8. The proliposomal powder of claim 1, comprising DPPC, DMPC, or a mixture of DPPC and DMPC. 9. The proliposomal powder of claim 8, wherein the mixture comprises at least 10% DMPC. 10. The proliposomal powder according to any one of claims 1 to 9, further comprising charged lipids. The group of claim 10 wherein the charged lipids comprise dimyristoyl phosphatidylglycerol (DMPG), diphosphalmitoyl phosphatidylglycerol (DPPG), dimyristoyl phosphatidic acid (DPPA) and stearylamine (SA). Selected proliposomal powder. The proliposomal powder according to any one of claims 1 to 11, wherein the active ingredient comprises a glucocorticosteroid. 13. The proliposomal powder of claim 12, wherein the active ingredient comprises a glucocorticosteroid that is esterified at position 21 with a fatty acid of at least 8 carbon atoms. The proliposomal powder of claim 13, wherein the active ingredient comprises a glucocorticosteroid that is esterified at position 21 with a fatty acid of at least 10 carbon atoms. The proliposomal powder of claim 14, wherein the active ingredient comprises glucocorticosteroid-21-palmitate. 16. The proliposomal powder of claim 15, wherein the active ingredient comprises roplefonide palmitate. The proliposomal powder of claim 1, wherein at least 50% of the powder consists of particles less than 10 microns in diameter. 18. The proliposomal powder of claim 17, wherein at least 50% of the powder consists of particles of 0.01-10 microns in diameter. The proliposomal powder of claim 18, wherein at least 50% of the powder consists of particles of 0.1-6 microns in diameter. 20. The proliposomal powder according to any one of claims 17 to 19 consisting of agglomerated particles. A pharmaceutical composition comprising the proliposomal powder of claim 1 and a pharmaceutically acceptable carrier. The pharmaceutical composition of claim 21, wherein the carrier is crystalline lactose monohydrate. The pharmaceutical composition of claim 21 or 22, wherein the carrier consists of particles of mass median particle size less than 10 microns or agglomerates of these particles. A dry powder inhaler device containing the proliposomal powder of claim 1. The dry powder inhaler device of claim 24, wherein the inhaler is a single dose inhaler. A predetermined biologically active compound comprising administering to a patient in need of such treatment a therapeutically effective amount of the proliposomal powder of claim 1 or the pharmaceutical composition of any one of claims 21-23. Method of treatment of patients in need of treatment. Use of the proliposomal powder of claim 1 in the preparation of a formulation for use in therapy.