WO2012074440A2

WO2012074440A2 - Pharmacological composition intended for intra-nasal administration with the aim of supplying a pharmacologically active component to the brain, and method for producing said composition

Info

Publication number: WO2012074440A2
Application number: PCT/RU2011/000944
Authority: WO
Inventors: Михаил Валентинович КОВАЛЬЧУК; Татьяна Николаевна БОРОДИНА; Илья Владимирович ДЕГТЕВ; Юлия Владимировна МОИСЕЕВА; Ирина Валерьевна МАРЧЕНКО; Наталия Валерьевна ГУЛЯЕВА; Татьяна Владимировна БУКРЕЕВА
Original assignee: Учреждение Российской Академии Наук Институт Кристаллографии Им. А.В.Шубникова Ран
Priority date: 2010-12-01
Filing date: 2011-11-30
Publication date: 2012-06-07
Also published as: RU2010149002A; WO2012074440A3; RU2475233C2

Abstract

The claimed group of inventions relates to the field of pharmacology and medicine. The pharmacological composition is intended for intra-nasal administration and enables an active substance to be supplied directly to the brain, by-passing the blood-brain barrier. The composition comprises a base container, a pharmacologically active component and a surface-modifying substance. Porous particles of an inorganic substance are used as the container, and the surface of the container is modified with surfactants, polymers or with a combination thereof. The containers made of an inorganic substance have a size in the range from 300 to 5000 nm. Inorganic substances selected from a group consisting of metal salts or oxides, for example calcium carbonate or titanium oxide, in the form of porous particles of a micron or sub-micron size, are used as the base container. For the surface-modifying substance, use is made of surfactants selected from polysorbates, or of polymers, selected from a group consisting of glycosaminoglycans, polypeptides, or combinations thereof. The method for producing the designated composition comprises producing the base container, sorbing the pharmacologically active substance in volumes of the pores in said container, and modifying the surface of said container. The base container is synthesized from inorganic substances in the form of porous particles of a micron or sub-micron size, and the surface of the designated particles of the base container is additionally modified with polymers selected from the group consisting of glycosaminoglycans or polypeptides, or with surfactants selected from the group consisting of polysorbates, or with combinations thereof.

Description

PHARMACOLOGICAL COMPOSITION

INTENDED FOR INTRASAL INTRODUCTION FOR THE DELIVERY OF THE PHARMACOLOGICALLY ACTIVE COMPONENT IN THE BRAIN,

AND METHOD FOR ITS PRODUCTION

The claimed group of inventions relates to the field of pharmacology and medicine and is intended for use in the treatment of diseases in which direct delivery of the active substance to the brain is required, bypassing the blood-brain barrier (BBB), for example, in the treatment of Parkinson's disease.

Targeted delivery of compounds to the brain is a major problem, fundamental both from the point of view of fundamental science and from the point of view of application in medicine. The unsolved problem significantly limits the development of modern approaches to the diagnosis and treatment of cerebral pathologies. Currently, the intranasal method of introducing a number of drugs (primarily of a peptide nature) is of great interest to researchers. It is believed that substances introduced intranasally are able to penetrate directly into the central nervous system (CNS) through the olfactory system, bypassing the BBB. The advantages of this route of administration are obvious: the introduction is non-invasive, and the bioavailable substance is rapidly absorbed and begins to act in a few minutes, which is important with emergency care.

However, not all substances can be effectively introduced in this way, so the expansion of the arsenal of substances administered intranasally is certainly relevant and necessary. Lipophilic compounds with a low molecular weight have good nasal bioavailability. Peptides and proteins with a molecular mass of more than 1000 Da are able to cross the nasal membrane only in small quantities. At the same time, a pronounced protective mechanism - cleansing of the ciliary epithelium - limits the time spent in the nasal cavity of both fluids and powders, the half-life of excretion is 15-30 minutes [1]. To reduce the rate of purification and increase the bioavailability of drugs, special “absorption enhancers” are used. Their action in

SUBSTITUTE SHEET (RULE 26) mainly consists in changing the permeability of the epithelial cell membrane by modifying the phospholipid bilayer, removing proteins from the membrane, or even removing the outer layer of the mucous membrane. The use of such auxiliary compounds can increase the bioavailability of many drugs, including peptides such as insulin.

Currently known intravenous administration of compositions based on nanoparticles containing the active substance, in which overcoming the BBB.

Known pharmacological composition intended for introduction into the brain of the active component with penetration through the BBB, containing a container substance, a pharmacologically active component and a surfactant (US patent N ° 61 17454, "Drag targeting to the nervous system by nanoparticles", IPC A61K 9/51 publ. 12.09.2000).

The disadvantages of the known composition are:

- the need to use some expensive components, namely cyanoacrylates, acrylamides, polylactates, alkylcyanoacrylates, polyanhydrides, etc .;

- polymer particles are mechanically unstable, which may complicate their storage and transportation;

- the release of the active component occurs due to the destruction of the polymer particles, thereby limiting the time of release of the active component to the time of destruction of the particles;

- One of the decay products of polyalkylcyanoacrylate particles is formaldehyde, which is toxic and has a strong negative effect on the central nervous system.

The objective of the proposed technical solution is to create a composition that uses an inexpensive base container, and the introduction of the composition is carried out intranasally.

The technical result is to obtain a composition in which inexpensive ingredients are used that do not have a harmful effect on the human body.

The technical task and result are achieved by the fact that in

SUBSTITUTE SHEET (RULE 26) a pharmacological composition intended for introduction into the brain of the active component, bypassing the BBB, containing a container base and a pharmacologically active component, porous particles of an inorganic substance were used as the container base, and the surface of the container was modified with surfactants, polymers, or a combination thereof. Inorganic matter containers have sizes ranging from hundreds of nanometers to tens of microns (100 to 5000 nm). Inorganic substances selected from the group of metal salts or metal oxides, for example, calcium carbonate or titanium oxide, in the form of porous micron or submicron particles can be used as the base container. As a surface modifying agent, surfactants selected from the group of polysorbates or polymers selected from the group consisting of glycosaminoglycans, polypeptides, or combinations thereof can be used.

A known method of obtaining a pharmacological composition intended for introduction into the brain of the active component with penetration through the BBB, including the production of nanoparticles with sorption of a pharmacologically active substance and the modification of their surface (US patent N ° 61 17454, "Drag targeting to the nervous system by nanoparticles", IPC A61K 9/51, publ. 12.09.2000). As the nanoparticles in the known method, synthetic polymer nanoparticles, for example, polyalkyl cyanoacrylate, albumin, polylactide, and other particles, are used.

The disadvantages of this method are:

- sterilization of polyalkyl cyanoacrylate particles is a difficult process. Autoclaving these particles leads to a significant increase in their size. The preparation of polyalkyl cyanoacrylate particles obtained as a result of lyophilization is very difficult to resuspend, which creates great difficulties in obtaining a suspension for injection;

- scaling of the method for producing polyalkyl cyanoacrylate particles can be difficult because centrifuging and dialysis processes carried out during the purification of polyalkyl cyanoacrylate particles during synthesis cause their aggregation;

- obtaining polylactide particles associated with the use of organic

SUBSTITUTE SHEET (RULE 26) solvents (acetone), which significantly limits the scope of their use, in particular for biomedical purposes;

- the production of polymer particles is associated with the use of laborious methods of polymer chemistry (emulsion polymerization, surface polymerization, etc.);

- in the synthesis of albumin particles, glutaraldehyde is used, which is a toxic substance that causes damage to the mucous membranes of the eyes, nasopharynx;

- with intranasal administration, the release of the active component from polyalkyl cyanoacrylate particles is difficult, because they are stable at acidic pH values (nasal pH is 4.5-6.0).

The objective of the proposed technical solution is to create a method that allows you to overcome these disadvantages of the known method.

The technical result is the development of a reliable and inexpensive way to obtain a pharmacological composition intended for intranasal administration into the brain of a pharmacologically active component.

The technical task and result are achieved by the fact that in the method for producing a pharmacological composition intended for delivery to the brain of a pharmacologically active component, including obtaining a container base, sorption of a pharmacologically active substance in its pore volumes and modification of its surface, the container base is synthesized from inorganic substances in the form of porous particles of micron or submicron size. Then, the surface of these particles is further modified with polymers selected from the group of glycosaminoglycans or polypeptides, or surfactants selected from the group of polysorbates, or combinations thereof. A container base can be obtained by mixing aqueous solutions containing Ca ²⁺ and Co ₃ ^{2 ′} ions, removing unbound ions by washing the precipitate and drying the formed microparticles. It is possible to obtain a container base by reacting titanium carbide powder with nitric acid. As the pharmacologically active substance of the composition, loperamide can be used. Modification of the surface of the base container can be performed as follows: microparticles

SUBSTITUTE SHEET (RULE 26) the substance used as the base container is placed in a solution of hyaluronic acid in an aqueous solution of NaCl, kept under stirring in this solution, the excess hyaluronic kistole is removed by washing with an aqueous solution of NaCl, and then dried.

The essence of the proposed group of inventions is illustrated by diagrams and photographs presented in the figures:

- figure 1: diagram of a method for producing a pharmacological composition;

- figure 2: image of a particle of calcium carbonate obtained using scanning electron microscopy;

- Fig.Z: images of particles of titanium dioxide obtained using transmission electron microscopy;

- figure 4: the results of the w ν / νο-test of particles of titanium dioxide, polybutyl cyanoacrylate and calcium carbonate loaded with loperamide:

* - significant differences from the control group at p <0.05

+ - significant differences from the control group at p <0, 1

p is the statistical significance of the result in the Kruskal-Wallis test;

- figure 5: the results of the in νί ^' νο test of unmodified particles of calcium carbonate loaded with loperamide, and the same particles coated with hyaluronic acid.

The scheme of operations. upon receipt of the proposed pharmacological composition is illustrated in figure 1.

The first operation is the synthesis of particles that will serve as a container that is a carrier of the active substance. As such particles, porous particles of calcium carbonate and titanium dioxide were formed.

Calcium carbonate

Formation of particles of calcium carbonate ^"- the subject of numerous scientific studies due to the widespread use of this material in the ^'various fields of industry, technology, medicine for the synthesis of porous SaS0 ₃ microparticles bsh used method [2], which is in direct mixing of solutions containing ions of Ca ^{2. +} and C0 ₃ ^{2 "} . In this case, first an amorphous precipitate is formed from CaC0 ₃ nanoparticles, and then in

SUBSTITUTE SHEET (RULE 26) The volume of their aggregates gives rise to microparticle growth centers. The quality of the obtained particles substantially depends on the experimental conditions, such as the type of salts used, their concentration, pH, temperature, the rate of mixing of the solutions, and the intensity of mixing of the reaction mixture. The particles had a fairly narrow size distribution (from 4 to 5 μm) and a mesoporous channel-like structure. The particle structure is visible in the photograph shown in figure 2. The CaCO3 powder was stored in a dry atmosphere, since in the presence of moisture, porous microspherulites formed by vaterite gradually pass into rhombohedral calcite crystals.

Titanium dioxide

Mesostructured particles of titanium dioxide were obtained by the interaction of titanium carbide powder with 5 M nitric acid [3]. According to transmission electron microscopy, the particles are formed by elongated nanocrystallites (Fig.Z). Using dynamic light scattering, it was determined that the average particle size is 1.5 microns.

Loperamide was adsorbed onto the obtained calcium carbonate and titanium dioxide particles by placing a weighed portion of these particles in an alcoholic solution of loperamide, followed by mixing of the suspension on a shaker. The amount of loperamide included in the containers was determined spectrophotometrically by the difference in intensity of the absorption peak of the supernatant at 259 nm before and after adsorption.

To modify the surface of the containers with hyaluronic acid, particles with included loperamide were placed in a solution of hyaluronic acid in an aqueous NaCl solution and kept under stirring with a shaker. Upon completion of the modification process, the particles were dried and stored as such.

For comparative purposes. tests were made polybutylcyanoacrylate particles with loperamide. Such particles were synthesized by emulsion polymerization of butyl cyanoacrylate in the presence of loperamide.

An example of a specific implementation of the method for producing the composition

Preformed inorganic nano or microparticles

SUBSTITUTE SHEET (RULE 26) incubated in an alcoholic solution of loperamide for 1-3 hours with stirring on a shaker (IKA-VIBRAX-VXR, IKA, Germany). After incubation, the particles were precipitated by centrifugation (ROTINA 38 / 38R, Hettich, Germany) and the supernatant was separated. The loperamide concentration in the supernatant was determined on a spectrophotometer (PerkenElmer Lambda 650, USA).

The obtained nano- or microcontainers were placed in an aqueous solution of a polymer, surfactant, or combinations thereof and incubated for 15-30 minutes with stirring on a shaker (IKA-VIBRAX-VXR, IKA, Germany). After incubation, the containers were besieged by centrifugation (ROTINA 38 / 38R, Hettich, Germany).

The obtained modified containers were dried for 2 hours at 50-60 ° C. Storage of the composition was carried out in a sealed tube at room temperature.

To test the effectiveness of the developed composition, an “in vivo” test was performed to evaluate the delivery of the active component to the brain. For this purpose, a formalin test using loperamide was used [4-6]. Rats were used as experimental animals. It is known that loperamide is not able to penetrate the BBB. Previously targeted delivery of loperamide to the central nervous system of the mouse was successfully accomplished by intravenous administration of polybutyl cyanoacrylate nanoparticles. In a comparative study, polybutyl cyanoacrylate particles containing loperamide were administered intranasally and data on the effectiveness of the administration of loperamide using porous calcium carbonate particles and titanium dioxide were compared with data for polybutyl cyanoacrylate particles.

Conclusions about the delivery of loperamide to the central nervous system of rodents were made on the basis of changes in their pain sensitivity. A suspension of particles loaded with loperamide was intranasally administered to Wistar rats weighing 200-250 g. Then, after subcutaneous administration of formaldehyde in the rat paw, the animal was placed in a transparent aquarium for observation. After a certain period of time, the behavior was recorded and the pain sensitivity of the rat was evaluated according to the point system. All time points were averaged over the group, data

SUBSTITUTE SHEET (RULE 26) processed using the STATISTICA-7 program. As a control, a solution of loperamide in a 5% particle-free glucose solution was used, as well as a suspension of the particles themselves without loperamide.

The scores in the control groups of animals did not significantly differ from each other, so they were combined and for further calculations took an average control.

All used particles loaded with loperamide reduced the pain sensitivity of rats (figure 4). This means that containers based on porous inorganic particles, as well as polybutyl cyanoacrylate particles, ensure the delivery of loperamide to the central nervous system of rats. The dynamics of reducing pain sensitivity is different for different types of particles. Particles of titanium dioxide, like polybutyl cyanoacrylate particles, are most effective at short durations, although polymer containers begin to act faster. Calcium carbonate particles showed good efficacy over long periods of action (42-60 min).

Modification of particles by a mucoadhesive compound led to an increase in the effectiveness of the containers at the beginning of the test and at large times (Fig. 5). When using calcium carbonate particles coated with hyaluronic acid, the pain sensitivity of rats is almost halved at the 6th minute and from the 48th to 66th minute. Considering that, at large times, CaC0 ₃ particles with loperamide act more efficiently than polybutyl cyanoacrylate particles, it can be concluded that containers based on calcium carbonate coated with hyaluronic acid from the studied systems are most effective for delivering loperamide to the central nervous system of rats.

Thus, the pharmacological composition and the developed method for its preparation make it possible to ensure the delivery of functional compounds to the brain during their intranasal administration. The simplicity of the composition in combination with the advantages of the intranasal route of administration of drugs provides the promise of the proposed system for medical applications.

Information sources -

1. Soane R.J., Frier M., Perkins A.S., Jones N.S., Davis S.S., Ilium L. // Int. J. Pharm. 1999. V. 178. P. 55-65.

SUBSTITUTE SHEET (RULE 26) 2, Volodkin DV, Petrov AI, Prevot M., Sukhorukov GB // Langmuir. 2004. V. 20. P. 3398-3406.

3. Shieh D.-L., Li Jr-Sh., Shieh M.-Jr, Lin J.-L. // Microporous and Mesoporous Materials. 2007. V. 98. P. 339-343.

4. Matthies B.K., Franklin K.B. // Pain. 1992. V. 51. P. 199-206.

5. Matthies B.K., Franklin K.B. // Behav. Brain Res. 1995. V. 67. P. 59-66.

6. Rosland J.H., Tjolsen A., Maehle B., Hole K. // Pain. 1990. V. 42. P. 235-242.

SUBSTITUTE SHEET (RULE 26)

Claims

CLAIM

1. A pharmacological composition intended for intranasal administration in order to deliver to the brain a pharmacologically active component, comprising a container base, a pharmacologically active component and a surface modifying substance, characterized in that porous particles of an inorganic substance are used as the container, and the surface of the container is modified with surfactants, polymers or a combination thereof.

2. The composition according to claim 1, characterized in that the containers of inorganic substances have a size in the range from 100 to 5000 nm.

3. The composition according to claim 1, characterized in that inorganic substances selected from the group of metal salts or metal oxides, for example, calcium carbonate or titanium oxide, in the form of porous micron or submicron particles are used as the base container.

4. The composition according to claim 1, characterized in that surfactants selected from the group of polysorbates or polymers selected from the group comprising glycosaminoglycans, polypeptides, or combinations thereof are used as surface-modifying substances.

5. A method of obtaining a pharmacological composition intended for intranasal administration in order to deliver a pharmacologically active component to the brain, including obtaining a container base, sorption of a pharmacologically active substance in its pore volumes, modification of its surface, characterized in that the container base is synthesized from inorganic substances in the form of porous particles of micron or submicron size, and the surface named. particles of the base container are further modified with polymers selected from the group of glycosaminoglycans or polypeptides, or surfactants selected from the group of polysorbates, or combinations thereof.

6. A method for producing a pharmacological composition according to claim 5, characterized in that the container base is prepared by mixing aqueous solutions containing Ca ²⁺ and Co ₃ ^{2 ~} ions, removing unbound ions by washing the precipitate and drying the formed microparticles.

SUBSTITUTE SHEET (RULE 26)

7. The method of obtaining the pharmacological composition according to claim 5, characterized in that the base container is obtained by the interaction of titanium carbide powder with nitric acid.

8. The method of obtaining the pharmacological composition according to claim 5, characterized in that loperamide is used as the pharmacologically active substance.

9. The method of obtaining the pharmacological composition according to claim 5, characterized in that to modify the surface of the base container, the microparticles of the substance are placed in a solution of hyaluronic acid in an aqueous solution of NaCl, kept under stirring in this solution, the excess hyaluronic acid is removed by washing with an aqueous solution of NaCl, and then dried.

SUBSTITUTE SHEET (RULE 26)