PL219255B1 - Polymer nanocapsule, and method for manufacturing and use thereof - Google Patents

Description

Description of the invention

The subject of the invention is new polymer nanocapsules of poly (n-butyl cyanoacrylate) obtained by interfacial polymerization with the use of oil-in-water microemulsions stabilized by sugar surfactants of the dicephalic and gemini type capable of encapsulating substances of a hydrophilic nature. Nanocapsules are intended for use in pharmaceutical formulations as carriers of active substances in cancer diagnostics and anti-cancer therapies.

The invention is also related to transparent, thermodynamic, stable and temperature-independent microemulsion systems stabilized with surfactants belonging to the group of sugar surfactants with the structure of the dicephalic -N-dodecyl-N, N-bis [(3-D-glucoheptonylamido) propyl] -amine structure (C12-DGHA) and gemini-type N, N'-bisdodecyl-N, N'-bis [(3-D-glucoheptonylamido) propyl] ethylenediamine (bis (C12-GHA)); description of the synthesis, respectively, in K. A. Wilk,

L. Syper, B. Barczyk, A. Sokołowski, B.W. Domagalska, J. Surf. Deterg. 3 (2000) 185-192 and K. A. Wilk, L. Syper, B.W. Domagalska, U. Komorek, I. Maliszewska, R. Gancarz, J. Surfact. Deterg. 5 (2002) 235-244.

One of the tasks of modern chemical technology is the search for such drug carriers and transport systems of biologically active compounds at the nano- and microscopic level, which would act in a selective and targeted manner, would limit the side effects of the drug, and at the same time would show the maximum therapeutic effect and ensure greater biological stability [R.C. Pinto, R.J. Neufeld, A.J. Ribeiro, F. Veiga, Nanomedicine; Nanotechnology, Biology, and Medicine 2 (2006) 8-21]. This task can be performed by nanoparticles: nonospheres and nanocapsules. The latter carriers deserve special attention, as they have a large loading capacity, allow long-term storage of the encapsulated compound and allow its greater accumulation at the target site, thus minimizing the side effects of the applied therapy.

The main methods of obtaining nanocapsules include methods using templating processes, in which drug carriers are prepared on the basis of templates, which are most often various types of micellar aggregates, such as micelles, emulsions, nanoemulsions, lyotropic liquid crystals, liposomes, etc. [T. Liua, Ch. Burgerb, B. Chu B., Prog. Polym. Sci. 28 (2003) 5-26]. Particularly interesting for application reasons is the use of microemulsions: transparent, thermodynamically stable mixtures of two immiscible liquids (polar and non-polar) stabilized with a surfactant or a mixture of surfactants (or surfactant and co-surfactant). One of the examples of templating processes leading to the production of nanocapsules is interfacial polymerization [H. Huang, E.E. Remsen, T. Kowalewski, K.L. Wooley, J. Am. Chem. Soc. 121 (1999) 3805-3806], which runs on the border of two immiscible phases [S.K. Karode, S.S. A.K. Suresh, R.A. Mashelkar, Chem Eng. Sci. 53 (1998) 2649-2663, V.V. Yashin, A.C. Balazs, J. Chem. Physics 121 (2004) 11440-11454]. In addition to the "unstirred interfacial polymerization" used for the preparation of artificial fibers, a similar process can be carried out at the interface of drops of one liquid dispersed in another (stirred interfacial polymerization) [W. Rempp, P. Merrill, Polymer Synthesis, Huthig and Wepf; (1991) pp. 280-287]. In this way, micro- and nanoparticles are obtained that are used as carriers for catalysts, pesticides, dyes or as drug carriers [J. Ji, R.F. Childs, M. Mehta, J. Membrane Sci. 192 (2001) 55-70]. The size of the obtained particles, their non-toxicity and easy biodegradation are extremely important issues during the preparation of nanocapsules.

Preparation of polystyrene nanocapsules using a nanoemulsion template is described by Chen et al. [Y. Chen, H. Liu, Z. Zhang, S. Wang, European Polymer Journal 43 (2007) 2848-2855]. Unfortunately, the use of this type of nanocapsule as drug carriers is impossible due to the toxicity of polystyrene.

The literature provides a description of the preparation of poly (alkyl cyanoacrylates) nanoparticles [US 4,329,332, US 4,489,055] useful for encapsulating hydrophilic substances, while there are few reports on such carriers for the encapsulation of hydrophobic compounds. Watnasirichaikul et al. [S. Watnasirichaikul, T. Rades, I.G. Tucker, N.M. Davies, Int. J. Pharmaceutics 235 (2002) 237-246] describes a method of producing nanocapsules based on a water-in-oil microemulsion stabilized with polysorbate 80 and sorbitan monooleate (3: 2 w / w), where a mixture was used as the oil phase. mono-, di- and triglycerides. The obtained nanoparticles are made of non-toxic and easily degraded polycyanoacrylates. Nanocapsules for enclosing hydrophilic insulin are described in [A. Graf, E. Ablinger, S. Peters, A. Zimmer, S. Hook, T. Rades, Int. J. Pharm. 350 (2008) 351-360]. The nanoparticles were obtained by interfacial polymerization of ethyl or butyl cyanoacrylate using as a template a water-in-oil microemulsion containing lecithin and isopropyl myristate.

Patent literature describes polycyanoacrylate nanoparticles obtained on the basis of an emulsion containing rose essence and stabilized polyoxyethylene sorbitan monolaurate - patent CN 101816911. Nanoparticles are obtained by interfacial polymerization lasting about 5 hours, the monomer is added in an amount from 0.6 to 1.6% by weight of the emulsion. The capsules obtained in this way have a size of approx. 100 nm and can be used in cosmetics or in fragrances for fabrics.

Another patent - CN 101092499 - discloses a method of preparing nanoparticles containing water-soluble active substances such as vitamin C and B, hyaluronic acid and its derivatives. Poly (ethyl or butyl cyanoacrylate) nanoparticles had a size of 100 to 500 nm and the encapsulated material showed high biocompatibility and the possibility of using it in cosmetics.

From US 2008182776, poly (cyanoacrylalkyl) nanoparticles are known. They are obtained by polymerization of nanoemulsions stabilized with non-ionic pluronic surfactants F127 or F68. These nanoparticles, compared to those obtained as a result of emulsion polymerization, were characterized by a higher degree of loading and encapsulation of the active substance.

The international application WO 2005/020933 describes a method of obtaining polymeric nanoparticles with sizes below 1000 nm, preferably from 1 to 400 nm. These particles are synthesized by emulsion polymerization.

Microemulsions are microheterophasic, transparent, thermodynamically stable mixtures of two immiscible liquids (polar and non-polar), stabilized with a surfactant or a mixture of surfactants (or surfactant and co-surfactant). After dissociation, the microemulsion components retain their chemical and physical separateness, and the droplet size of the dispersed phase (5-100 nm) ensures system stability. According to the invention, for the production of new microemulsion systems, non-ionic sugar surfactants are used, produced by simple syntheses with the use of renewable raw materials. These surfactants are biodegradable, meet the requirements of green chemistry and are more environmentally friendly than conventional surfactants. The proposed microemulsions can be used to solubilize both hydrophilic compounds (in the case of water-in-oil microemulsions, w / o) and hydrophobic (oil-in-water microemulsions, o / w) and amphiphilic (two-continuous microemulsions). ). They can be used in pharmaceutical and cosmetic formulations. In the food industry, as a medium in the polymerization reaction, in environmental protection, e.g. for remediation of contaminated soil and for the preparation of polymer nanocapsules.

In the scientific and patent literature, reports have appeared on the production and use of various types of microemulsions.

Otto Glatter et al., Sugar-Ester Nonionic Microemulsion: Structural Characterization, J. Colloid Interface Sci. 241 (2001) 215-225 and Monzer Faun, Properties of microemulsions with sugar surfactants and peppermint oil, Colloid Polym Sci. 287 (2009) 899-910 discloses that sugar surfactants can form microemulsions in the surfactant / co-surfactant / oil / water system.

From the patent literature there are known oil-in-water microemulsions or bi-continuous microemulsions containing a mixture of surfactants (in an amount up to 15% by weight) useful as a carrier for the administration of active compounds with low water solubility (such as lecithin, sphingolipids and galactolipids) - application Patent No. P325825.

Another 1999 patent application (PCT / AUS99 / 17319) discloses a fragrance-containing water-in-oil microemulsion that is miscible with an aqueous fabric softener base composition useful for the delivery of a perfume.

Polish patent 201177 (P363307) discloses an edible water-in-oil microemulsion which comprises oil, a surfactant, a co-surfactant and an aqueous phase containing water and optionally water-soluble ingredients. The content of the oil phase in the present system ranges from 25 to 97% by weight of the diglycerides, and the molar ratio of the water phase to the surfactant ranges from 5 to 70. The patent also discloses a method for producing such a microemulsion.

PL 219 255 B1

GB 2297759 describes microemulsions that can be used in the food industry. They are stabilized with polyglycerol esters of unsaturated fatty acids containing from 12 to 24 carbon atoms in the hydrophobic chain. The weight ratio of surfactant to water is less than 5: 1, the preferred amount of water in the microemulsion does not exceed 2% by weight, which makes these systems not very effective at solubilizing water.

In turn, the patent no. US 2007003581 relates to microemulsions with potential use as cosmetic and pharmaceutical formulations, which do not irritate the skin and do not show toxicity. The described microemulsions can be used to transfer active compounds (e.g. vitamins, antioxidants). These systems are formed with co-surfactants, such as polyglycerols or various monoesters, and are stabilized with polyethylene glycol-based emulsifiers. The referenced patent shows a pseudo-ternary phase diagram which is helpful in delineating the areas of occurrence of microemulsions.

From the patent literature there are also known w / o microemulsions for transporting flavor and aroma compounds in food products (international application WO 9962357). In preferred systems, the molar ratio of water to surfactant is less than 10.

In the literature, there are no known methods of producing polymer nanocapsules obtained by interfacial polymerization with the use of oil-in-water (o / w) microemulsion, stabilized with sugar surfactant of the dicephalic or gemini type.

The present invention relates to a polymer poly (alkyl cyanoacrylate) nanocapsule containing a liquid core of an oil-in-water microemulsion, characterized by being stabilized with a sugar surfactant selected from the group consisting of: N-dodecyl-N, N-bis [(3-D -glucoheptonylamido) propyl] amine or N, N'-bisdodecy N, N'-bis [(3-D-glucoheptonyl amido) propyl] ethylenediamine.

Preferably, the nanocapsule according to the invention further comprises an active substance, especially an anti-cancer active substance.

Another object of the invention is a process for the production of the above nanocapsule from poly (alkyl cyanoacrylate), characterized in that:

a. an emulsifier is obtained by mixing the sugar surfactant and iso-butanol in a weight ratio of 1: 1, using a sugar surfactant selected from the group consisting of: N-dodecylN, N'-bis [(3-D-gIucoheptonylamido) propyl] amine or N, N'-bisdodecyl-N, N'-bis [(3-D-gIukoheptonyI amido) propyl] ethylenediamine,

b. to the obtained emulsifier is added from 1 to 70% by weight of the oil phase,

c. an aqueous phase is added to the resulting mixture in an amount of 1 to 99% by weight and left at a temperature of 20 to 50 ° C to form a microemulsion,

d. the obtained microemulsion templates are mixed with a 1: 3 by volume solution of alkyl cyanoacrylate in chloroform to obtain stable nanocapsules.

In the above process, the surfactant is preferably used in an amount of 1-60 wt%, the oil phase in an amount of 0.1-20 wt%, more preferably 0.1-5 wt% is used. oil and 1-5% surfactant, according to the pseudo-ternary diagrams.

In the process according to the invention, diethyl glycol monoethyl ether is preferably used as the oil phase.

In the method according to the invention, preferably after mixing all components, the sample is equilibrated for at least 24 h at room temperature.

In the process of the invention, preferably an alkyl, more preferably ethyl or n-butyl cyanoacrylate monomer is used.

In the method of the invention, the monomer is preferably used in an amount of from 1 to 10 µM per ml of microemulsion, more preferably from 5 to 8 µM of monomer per ml of microemulsion.

In the process of the invention, preferably the interfacial polymerization process is carried out at a temperature below 10 ° C, more preferably 5 ° C.

In the process of the invention, preferably the interfacial polymerization process is carried out for at least 6 hours, more preferably 4 hours.

The interfacial polymerization process is preferably carried out under continuous agitation, with a mixing speed of from 500 to 1200 rpm, more preferably 1200 rpm.

The subject of the invention is explained in the examples of the preparation of nanocapsules on microemulsion templates stabilized with the sugar surfactant N-dodecyl-N, N-bis [(3-D-glucoheptonylamido) propyl] amine or N, N'-bisdodecyl-N, N'-bis [( 3-D-glucoheptonyl amido) propyl] ethylenediamine and

1 shows pseudo-ternary phase diagrams on the basis of which the areas of occurrence of microemulsions are determined and where: ME - microemulsion, o / w - oil-in-water. w / o - water-in-oil, bc - two-continuous system. Figures 2, 3, 4 show, respectively: the measurements of the hydrodynamic diameter of the capsules obtained by the dynamic light scattering (DLS) technique and the morphology of nanocarriers obtained using the scanning electron microscope, SEM and the atomic force microscope, AFM.

Figures 5 and 6 and Table 4 show the medical potential of the invention (exemplary results for nanocapsules). Polymerizable nanocapsules based on microemulsions containing sugar surfactants show low haemolytic activity. It is not affected by the presence of the photosensitizer.

Empty nanocapsules show low cytotoxicity: the survival of unexposed wt MCF-7 cells is 96.6%, 84% and 76.4%, respectively, ie the "dark toxicity" effect is low.

After irradiation of cells incubated with an empty nanocarrier, the survival of wt MCF-7 decreased to 23% and approx. 4% in the presence of both empty carriers and those loaded with cyanines. The resulting nanocarriers may be useful for the transport of hydrophobic photosensitizing agents through the circulation.

The results of flow cytofluorimetry (FACS) studies after incubation of MCF-7WT cells, shown in Fig. 8, prove that the nanocapsules of the invention deliver cyanine to cells much more efficiently (Fig. 8d) compared to free drug (Fig. 8). 8 b and 8c). Figure 8a shows the negative control - MCF-7 / WT cell population without nanocapsules. Figures 8d - 9g also show histograms with mean fluorescence intensity in cells incubated with free cyanine (Figure 8d), cyanine capsules IR-780 (8e) and IR-786 (Figure 8f), and nanocapsules without cyanine (Figure 8d) 8g).

P r z k ł a d I

0.25 g of N-dodecyl-N, N-bis [(3-D-glucoheptonylamido) propyl] amine and 0.25 g of iso-butanol are mixed. To the so prepared emulsifier was added 0.5 g of diethyl glycol monoethyl ether (DGME) and 9.0 g of water. It is left at 25 ° C for 24 hours for equilibrium. After this time, a transparent, thermodynamically stable microemulsion of the o / w type is obtained with an oil content of 5% and an emulsifier content of 5% and a droplet diameter of 6.8 nm. The microemulsion is cooled to 5 ° C, and n-butyl cyanoacrylate dissolved in chloroform (1: 3, v / v) is introduced into it in such an amount that there are 5 µl of monomer per milliliter of microemulsion. The obtained nanocapsules have a diameter of 340.7 nm, a polydispersity index of 0.127 and a zeta potential of -29.5 mV.

P r z x l a d II

0.5 g of N-dodecyl-N, N-bis [(3-D-glucoheptonylamido) propyl] amine and 0.5 g of isobutanol are mixed. To the so prepared emulsifier is added 0.5 g of diethyl glycol monoethyl ether (DGME) and 8.5 g of water. It is left at 25 ° C for 24 hours for equilibrium. After this time, a transparent, thermodynamically stable microemulsion of the o / w type with an oil content of 5% and an emulsifier content of 10% and a droplet diameter of 7.0 nm is obtained. The microemulsion is cooled to 5 ° C, and n-butyl cyanoacrylate dissolved in chloroform (1: 3, v / v) is introduced into it in such an amount that there are 5 µl of monomer per milliliter of microemulsion. The obtained nanocapsules have a diameter of 352.4 nm, a polydispersity index of 0.108 and a zeta potential of -29.3 mV.

P r x l a d III

0.25 g of N-dodecyl-N, N-bis [(3-D-glucoheptonylamido) propyl] amine and 0.25 g of isobutanol are mixed. To the so prepared emulsifier was added 0.5 g of diethyl glycol monoethyl ether (DGME) and 9.0 g of water. It is left at 25 ° C for 24 hours for equilibrium. After this time, a transparent, thermodynamically stable microemulsion of the o / w type is obtained with an oil content of 5% and an emulsifier content of 5% and a droplet diameter of 6.8 nm. The microemulsion is cooled to 5 ° C, and n-butyl cyanoacrylate dissolved in chloroform (1: 3, v / v) is introduced into it in such an amount that there are 8 µl of monomer for each milliliter of microemulsion. The obtained nanocapsules have a diameter of 352.5 nm, polydispersity index 0.118 and zeta potential equal to -29.8 mV.

P r x l a d IV

0.25 g of N, N'-bisdodecyl-N, N'-bis [(3-D-glucoheptonylamido) propyl] ethylenediamine and 0.25 g of iso-butanol are mixed. To the thus prepared emulsifier, 0.5 g of diethyl glycol monoethyl ether (DGME) and 9.0 g of water are added. It is left at 25 ° C for 24 hours for the purpose

Equilibrium. After this time, a transparent, thermodynamically stable o / w microemulsion with an oil content of 5% and an emulsifier content of 5% and a droplet diameter of 3.9 nm is obtained. The microemulsion is cooled to 5 ° C, and n-butyl cyanoacrylate dissolved in chloroform (ratio 1: 3, v / v) is introduced into it in such an amount that each milliliter of microemulsion contains 5 µM of monomer. The obtained nanocapsules have a diameter of 214.8 nm, polydispersity index 0.147 and zeta potential equal to -31.2 mV.

P r z k ł a d V

0.25 g of N-dodecyl-N, N-bis [(3-D-glucoheptonylamido) propyl] amine and 0.25 g of iso-butanol are mixed. To the so prepared emulsifier was added 0.5 g of diethyl glycol monoethyl ether (DGME) and 9.0 g of water. It is left at 25 ° C for 24 hours for equilibrium. After this time, a transparent, thermodynamically stable microemulsion of the o / w type is obtained with an oil content of 5% and an emulsifier content of 5% and a droplet diameter of 6.8 nm. The microemulsion is cooled to 5 ° C, and ethyl cyanoacrylate dissolved in chloroform (ratio 1: 3, v / v) is introduced into it in such an amount that for each milliliter of microemulsion there is 5 μΐ of monomer. The obtained nanocapsules with a diameter of 321.0 nm, polydispersity index 0.113 and zeta potential equal to -26.3 mV.

P r x l a d VI

0.25 g of W, W′-bisdodecyl-W, W′-bis [(3-D-glucoheptonylamido) propyl] ethylenediamine and 0.25 g of iso-butanol are mixed. To the so prepared emulsifier was added 0.5 g of diethyl glycol monoethyl ether (DGME) and 9.0 g of water. It is left at 25 ° C for 24 hours for equilibrium. After this time, a transparent, thermodynamically stable o / w microemulsion with an oil content of 5% and an emulsifier content of 5% and a droplet diameter of 3.9 nm is obtained. The microemulsion is cooled to 5 ° C, and ethyl cyanoacrylate dissolved in chloroform (ratio 1: 3, v / v) is introduced into it in such an amount that for each milliliter of microemulsion there are 5 μl of monomer. The obtained nanocapsules have a diameter of 198.2 nm, polydispersity index 0.109 and zeta potential equal to -27.5 mV.

P r o x l a d VII

0.25 g of N-dodecyl-N, N-bis [(3-D-glucoheptonylamido) propyl] amine and 0.25 g of iso-butanol are mixed. To the so prepared emulsifier was added 0.5 g of diethyl glycol monoethyl ether (DGME) and 9.0 g of water. It is left at 25 ° C for 24 hours for equilibrium. After this time, a transparent, thermodynamically stable microemulsion of the o / w type is obtained with an oil content of 5% and an emulsifier content of 5% and a droplet diameter of 6.8 nm. To the thus prepared microemulsion, cyanine IR-780 is added in such an amount that it does not dissolve completely, and the system is stirred for 4 hours. Excess cyanide is removed through a 0.2 µm syringe filter. The miltroemulsion with the solubilized photosensitizer is cooled to 5 ° C, and ethyl cyanoacrylate dissolved in chloroform (ratio 1: 3, v / v) is introduced into it in such a quantity that for each milliliter of microemulsion there are 5 μl of monomer. The obtained nanocapsules were loaded with IR-780 cyanine with a diameter of 333.0 nm, polydispersity index 0.078, zeta potential of -29.6 mV and cyanine encapsulation degree of 91.9%.

P r x l a d VIII

0.25 g of N, N'-bisdodecyl-N, N'-bis [(3-D-glucoheptonylamido) propyl] ethylenediamine and 0.25 g of iso-butanol are mixed. To the thus prepared emulsifier, 0.5 g of diethyl glycol monoethyl ether (DGME) and 9.0 g of water are added. It is left at 25 ° C for 24 hours for equilibrium. After this time, a transparent, thermodynamically stable o / w microemulsion with an oil content of 5% and an emulsifier content of 5% and a droplet diameter of 3.9 nm is obtained. To the thus prepared microemulsion, cyanine IR-780 is added in such an amount that it does not dissolve completely, and the system is stirred for 4 hours. Excess cyanide is removed through a 0.2 µm syringe filter. The microemulsion with the solubilized photosensitizer is cooled to 5 ° C, and ethyl cyanoacrylate dissolved in chloroform (1: 3, v / v) is introduced into it in such an amount that for each milliliter of microemulsion there are 5 μl of monomer. The obtained nanocapsules were loaded with cyanine IR-780 with a diameter of 234.3 nm, a polydispersity index of 0.224, a zeta potential of -31.0 mV and a cyanine encapsulation degree of 92.6%.

P r x l a d IX

0.25 g of N-dodecyl-N, N-bis [(3-D-glucoheptonylamido) propyl] amine and 0.25 g of iso-butanol are mixed. To the so prepared emulsifier was added 0.5 g of diethyl glycol monoethyl ether (DGME) and 9.0 g of water. It is left at 25 ° C for 24 hours for equilibrium.

PL 219 255 B1

After this time, a transparent, thermodynamically stable microemulsion of the o / w type is obtained with an oil content of 5% and an emulsifier content of 5% and a droplet diameter of 6.8 nm. To the thus prepared microemulsion, cyanine IR-786 is added in such an amount that it does not dissolve completely, and the system is stirred for 4 hours. Excess cyanide is removed through a 0.2 µm syringe filter. The microemulsion with the solubilized photosensitizer is cooled to 5 ° C, and ethyl cyanoacrylate dissolved in chloroform (1: 3, v / v) is introduced into it in such an amount that for each milliliter of microemulsion there is 5 μΐ of monomer. The obtained nanocapsules loaded with cyanine IR-786 with a diameter of 362.3 nm, polydispersity index 0.139, zeta potential equal to -29.8 mV and cyanine encapsulation degree equal to 87.6%.

P r z k ł a d X

0.25 g of N, N'-bisdodecyl-N, N'-bis [(3-D-glucoheptonylamido) propyl] ethylenediamine and 0.25 g of iso-butanol are mixed. To the so prepared emulsifier was added 0.5 g of diethyl glycol monoethyl ether (DGME) and 9.0 g of water. It is left at 25 ° C for 24 hours for equilibration. After this time, a transparent, thermodynamically stable o / w microemulsion with an oil content of 5% and an emulsifier content of 5% and a droplet diameter of 3.9 nm is obtained. To the thus prepared microemulsion, cyanine IR-786 is added in such an amount that it does not dissolve completely, and the system is stirred for 4 hours. Excess cyanide is removed through a 0.2 µm syringe filter. The microemulsion with the solubilized photosensitizer is cooled to 5 ° C, and ethyl cyanoacrylate dissolved in chloroform (ratio 1: 3, v / v) is introduced into it in such an amount that for each milliliter of microemulsion there is 5 μ of monomer. The obtained nanocapsules loaded with cyanine IR-786 with a diameter of 220.7 nm, polydispersity index 0.152, zeta potential equal to -31.3 mV and cyanine encapsulation degree equal to 89.0%.

The characteristics of the microemulsion (hydrodynamic diameter, DH, polydispersity coefficient, Pdl, ratio of peak 1 and 3 intensity in the fluorescence spectrum of pyrene solubilized in I1 / I3 microemulsions are summarized in Table 1 below.

T a b e l a 1

Selected properties of microemulsion templates formed in the following systems: sugar surfactant: iso-butanol / DGME / water

percentage type ME ^a) 25 ° C 40 ° C 55 ° C surfactant + co-surfactant Oil water Dh (nm) ^b) Pdl ^c) I1 / l3 ^d) Dh (nm) ^b) Pdl ^c) Dh (nm) ^b) Pdl ^c) C12-DGHA: iso-butanol / DGME / water 5 5 90 that 6.8 0.163 1.39 6.2 0.251 6.2 0.358 10 5 85 that 7.0 0.182 1.38 6.0 0.261 6.1 0.341 15 5 80 that 7.4 0.126 1.38 7.1 0.396 6.7 0.452 bis (C12-GHA): iso-butanol / DGME / water 5 5 90 that 3.9 0.113 nb 3.7 0.192 3.5 0.210 10 5 85 that 4.5 0.123 nb 4.2 0.211 4.0 0.241 15 5 80 that 5.3 0.112 nb 5.1 0.246 4.8 0.341

^a) type of microemulsion determined on the basis of conductometric measurements, ^b) DH - hydrodynamic diameter measured by DLS technique, ^c) PdI - polydispersity index; nb - not tested

Table 2 shows how the reaction time, monomer concentration and pH influenced the nanocapsule preparation process, while Table 3 shows the characteristics of the obtained nanocapsules empty and loaded with cyanine IR-780 and cyanine IR-786. Table 4 presents the medical aspect of the invention - the hemolytic potential of the obtained nanocapsules. As temlates in the presented cases, microemulsions containing 5% emulsifier and 5% oil phase were used.

PL 219 255 B1

T a b e l a 2

Influence of the process time, monomer concentration and pH on the size of nanocapsules and polymerization efficiency

microemulsion / templat response time amount of monomer (gl / ml] PH Dh [nm] ^a) Dh2 [nm] ^a) Pdl ^b) In [%] ω Ό about LU ABOUT Q o c ro -1— < = 3 _Q ABOUT N < AND ABOUT Q CM about Twenty minutes 5.0 2.5 227.3 2245.1 0.810 - Thirty minutes 5.0 2.5 250.6 2193.4 0.769 62.1 60 minutes 5.0 2.5 274.1 1843.2 0.654 74.7 2 hours 5.0 2.5 280.6 1695.9 0.421 89.5 3 hours 5.0 2.5 300.6 - 0.173 92.9 4 hours 1.0 2.5 272.3 - 0.231 90.3 3.5 2.5 289.4 - 0.198 95.4 5.0 2.5 307.9 - 0.114 98.6 8.0 2.5 352.5 - 0.118 99.0 14.0 2.5 > 1 gm - 0.983 99.1 5.0 5.0 336.3 - 0.238 98.7 5.0 6.0 382.7 - 0.531 97.6 5.0 7.0 - - - 98.5 5.0 8.0 - - - 97.9 5 hours 5.0 2.5 305.5 - 0.136 98.7 6 hours 5.0 2.5 310.3 - 0.126 99.0 8 hours V 2.5 312.8 - 0.129 99.2

^a DH - hydrodynamic diameter and ^b PdI - polydispersity index obtained from DLS measurements

Table 3 presents the parameters of empty and loaded photosensitizing poly (n-butyl cyanoacrylate) capsules obtained from microemulsions stabilized with sugar surfactants

T a b e l a 3

Size (DH), polydispersity (PdI) and zeta potential (ξ) of poly (n-butyl cyanoacrylate) capsules empty and loaded with photosensitizers, obtained from microemulsions stabilized with sugar surfactants

microemul- sja / templat Nanocapsules empty loaded with IR-780 loaded with LR-786 system Dh [nm] PdI ξ [mv] system Dh [nm] PdI ξ [mV] EE% ^a system DH [nm] PdI ξ [mV] EE% ^a C12-DGHA: iso-butanol / DGME / water K190 340.7 0.127 -29.5 K193 333.0 0.078 -29.6 91.9 K196 362.3 0.139 -29.8 87.6 bis (C12-GHA): iso-butanol / DGME / water K191 214.8 0.147 -31.2 K194 234.3 0.224 -31.0 92.6 K197 220.7 0.152 -31.3 89.0

^a Degree of encapsulation (EE%)

PL 219 255 B1

T a b e l a 4

Hemolytic potential of poly (n-butyl cyanoacrylate) nanocapsules based on microemulsions stabilized by sugar surfactants (designations as in Table 3 above)

Nanocapsules 0.5 mg / ml Sample volume (μ!) PBS μΐ Concentration μg / ml The degree of dilution in the RBC suspension A540 H. (%) EMPTY 1 49 2 1: 250 0.047 2.3 KP * 2.5 47.5 5 1: 100 0.050 2.6 5 45 10 1:50 0.054 3.1 15 35 thirty 1:17 0.057 3.5 twenty thirty 40 1: 12.5 0.066 3.8 40 10 80 1: 6.25 0.069 4.2 FULL - with the IR-780 K193 1 49 2 1: 250 0.062 4.2 2.5 47.5 5 1: 100 0.068 4.8 5 45 10 1:50 0.071 5.3 15 35 thirty 1:17 0.076 5.9 twenty thirty 40 1: 1: 12.5 0.078 6.2 40 10 80 1: 6.25 0.080 6.5 K194 1 49 2 1: 250 0.049 2.5 2.5 47.5 5 1: 100 0.052 2.9 5 45 10 1:50 0.058 3.6 15 35 thirty 1:17 0.061 4.1 twenty thirty 40 1: 12.5 0.063 4.4 40 10 80 1: 6.25 0.067 4.8

* KP nanocapsule empty - analogue of K192 without cyanine

Claims (11)

Patent claims A polymeric poly (alkyl cyanoacrylate) nanocapsule containing a liquid core of an oil-in-water microerulsion, stabilized by a sugar surfactant selected from the group consisting of: N-dodecyl-N, N-bis [(3-D-glucoheptonylamido) ) propyl] amine or N, N'-bisdodecyl-N, N'-bis [{3-D-glucoheptonyl amido) propyl] ethylenediamine. 2. The nanocapsule according to claim 1 3. A composition according to claim 1, characterized in that it further comprises an active substance, especially an anti-cancer substance. 3. A method for producing a poly (alkyl cyanoacrylate) nanocapsule according to claim 1; 1, characterized in that: a. obtaining an emulsifier by mixing sugar surfactant and iso-butanol in a weight ratio of 1: 1, using a sugar surfactant selected from the group consisting of; N-dodecyl-N, N-bis [(3-D-glucoheptonylamido) propyl] amine or N, N'-bisdodecyl-N, N'-bis [(3-D-glucoheptonyl amido) propyl] ethylenediamine, b. to the obtained emulsifier is added from 1 to 70% by weight of the oil phase, c. an aqueous phase is added to the resulting mixture in an amount of 1 to 99% by weight and left at a temperature of 20 to 50 ° C to form a microemulsion, d. the obtained microemulsion templates are mixed with a 1: 3 by volume solution of alkyl cyanoacrylate in chloroform to obtain stable nanocapsules. PL 219 255 B1 4. The method according to p. 3. The process of claim 3, characterized in that the surfactant is used in an amount of 1 - 60 wt.%, The oil phase in an amount of 0.1 - 20 wt.%, Preferably 0.1 - 5 wt.% Is used. oil and 1-5% surfactant, according to the pseudo-ternary diagrams. 5. The method according to p. The process of claim 3, characterized in that diethyl glycol monoethyl ether is used as the oil phase. 6. The method according to p. 3. The method according to claim 3, characterized in that after mixing all components, the sample is equilibrated for at least 24 h at room temperature. 7. The method according to p. The monomer according to claim 3, wherein the monomer is an alkyl, preferably ethyl or n-butyl cyanoacrylate. 8. The method according to p. The process of claim 3, wherein the monomer is used in an amount of from 1 to 10 µΐ per ml of microemulsion, preferably from 5 to 8 µΐ of monomer per ml of microemulsion. 9. The method according to p. The process of claim 3, characterized in that the interfacial polymerization process is carried out at a temperature below 10 ° C, preferably 5 ° C. 10. The method according to p. The process of claim 3, characterized in that the interfacial polymerization process is carried out for at least 6 h, preferably 4 h. 11. The method according to p. 3. The process of claim 3, wherein the interfacial polymerization process is carried out under continuous agitation at a mixing speed of 500 to 1200 rpm, preferably 1200 rpm.