Celecoxib nanocrystal, process for producing celecoxib nanocrystal, pharmaceutical composition, use of a celecoxib nanocrystal, and method of treatment for relief of pain- and inflammation-related conditions

Abstract

The present invention relates to a technology that aims to optimise the onset of action of a molecule having analgesic and anti-inflammatory properties, specifically celecoxib or derivatives thereof. The approach involves the production of nanocrystals of celecoxib or derivatives thereof stabilised with a low-viscosity polymer for a faster onset of action of the active ingredient, as well as the production process therefor, pharmaceutical compositions, uses and treatment methods comprising same.

Description

Description Title of the Invention: CELECOXIB NANOCRYSTAL, PROCESS FOR PRODUCING CELECOXIB NANOCRYSTAL, PHARMACEUTICAL COMPOSITION, USE OF A CELECOXIB NANOCRYSTAL AND TREATMENT METHOD FOR RELIEF OF PAIN AND INFLAMMATION-RELATED CONDITIONS Field of Invention

[0001] The present invention relates to celecoxib nanocrystals or its derivatives stabilized with low-viscosity polymers, for a faster onset of action of this anti-inflammatory drug, as well as its production process, pharmaceutical compositions, uses and treatment methods comprising the same. Background of the Invention

[0002] Celecoxib (CLX) is a nonsteroidal anti-inflammatory drug (NSAID), a COX-2 inhibitor, whose mechanism of action is to reduce the production of substances associated with pain and inflammation in the human body. Therefore, it has been indicated to relieve pain, tenderness, swelling, and stiffness caused by osteoarthritis (arthritis caused by breakdown of the joint lining), rheumatoid arthritis (arthritis caused by swelling of the joint lining), as well as to treat juvenile rheumatoid arthritis (a type of arthritis that affects children) in children 2 years of age and older.

[0003] In many cases, faster pharmacological action may be desirable to reduce patient suffering and improve treatment adherence (Baryakova, TH, Pogostin, BH, Langer, R. et al. Overcoming barriers to patient adherence: the case for developing innovative drug delivery systems. Nat Rev Drug Discov 22, 387-409 (2023)). Considering the different routes of administration, the intravenous route offers 100% bioavailability and immediate action and, therefore, would be the most efficient for treatments requiring the fastest pharmacological action.

[0004] However, the most commonly used route of administration is oral, as it does not require sterility, is safer, more convenient, and allows for self-administration (Alqahtani MS, Kazi M, Alsenaidy MA and Ahmad MZ (2021) Advances in Oral Drug Delivery. Front. Pharmacol. 12:618411.).

[0005] Typically, in the state of the art, when rapid oral action is desired, liquid pharmaceutical forms based on solutions are used, which eliminate the dissolution step and are readily absorbed, such as syrups and drops. However, solution pharmaceutical forms require that the active ingredient be readily soluble in aqueous medium, as is the case with molecules with Biopharmaceutical Classification (BCS) I or III. This does not apply to molecules with BCS II, which are poorly soluble in aqueous medium, as is the case with CLX, which exhibits low solubility and high permeability, with variable bioavailability due to its in vivo dissolution. It is also a weak acid with a pKa of 11.1.

[0006] Numerous alternatives have been researched to provide the appropriate application of the drug, for example, the use of nanocrystals.

[0007] Nanocrystals are non-matrix particulate drug systems on the nanoscale, stabilized by surfactants, polymers, or a mixture of both. Generally, the size of these particles is less than 1000 nm, typically between 100 and 500 nm. The term nanocrystal refers to the stable nanometric structure of the active ingredient, and can be applied to either the liquid or solid external phase. When in liquid form, the terms nanosuspension or nanodispersion can also be used.

[0008] The most widely used technology for producing nanocrystals is called top-down and consists of initially starting with large particles and then, by supplying high energy, causing breaks in their structure, resulting in smaller particles. This technique is widely used mainly because it does not involve organic solvents. Its most commonly used forms are wet milling and high-pressure homogenization (Raida Al-Kassas, Mahima Bansal, John Shaw, Nanosizing techniques for improving bioavailability of drugs, Journal of Controlled Release, Volume 260, 2017, Pages 202-212, ISSN 0168-3659), but other methodologies for solid fragmentation can also be cited, such as sonication, cavitation, shearing, compression, impact, and cutting.

[0009] According to the present invention, the top-down method includes wet or dry grinding with a ball mill, high-energy grinding, high-pressure homogenization, and similar comminution processes. In the case of the high-pressure homogenizer, homogenization is carried out in a pressure range of 100-2000 bar, using 1 to 20 cycles.

[0010] For the production of nanocrystals using the wet ball mill technique, the ratio between celecoxib and the grinding beads is 1:1 to 1:10 (active ingredient: beads), the rotation speed is 1000-1800 RPM, and the processing time is 60-240 minutes.

[0011] In contrast, there is another technology called bottom-up, which consists of starting with the molecular (soluble) form and forming the particles in the medium so that they are structured in situ. Both technologies reach nanometric sizes and favor increased solubility, improved dissolution rate, absorption, and action of the active ingredient. Furthermore, the following advantages and distinctions of the top-down technique compared to the bottom-up technique in the production of nanocrystals can be highlighted: Size Control

[0012] Top-down: Offers precise control over the size of the nanocrystals, allowing for specific adjustments and the production of crystals of uniform size.

[0013] Bottom-up synthesis can result in variations in nanocrystal size due to the complexity of synthesis from molecules or atoms. Efficiency in Mass Production

[0014] Top-down: This is often more efficient in the mass production of nanocrystals, as it involves reducing larger materials to the desired size.

[0015] Bottom-up: It can be challenging and less efficient when trying to produce large volumes of nanocrystals due to the synthesis of individual molecules. Scale and Reproducibility

[0016] Top-down: It is easier to scale up for large-scale production, and the approach generally offers good reproducibility in the properties of the nanocrystals.

[0017] Bottom-up: It can be more complicated to scale up, and reproducibility can be a challenge due to sensitivity to synthesis conditions. Standardization and Uniformity

[0018] Top-down: Allows for greater standardization and uniformity in the nanocrystals produced, which is crucial in many applications.

[0019] Bottom-up: Uniformity can be a challenge, especially when the synthesis involves the self-organization of molecules. Versatility in Starting Materials

[0020] Top-down: It can be applied to a variety of starting materials, allowing the production of nanocrystals from different sources.

[0021] Bottom-up: May be more limited by the availability of specific precursors and ideal synthesis conditions. Less Material Waste

[0022] Top-down: Generally involves less material waste, as it starts with larger materials that can be processed efficiently.

[0023] Bottom-up: This can generate unwanted byproducts, resulting in greater material waste during the synthesis process.

[0024] The state of the art teaches some nanometric systems for drugs that exhibit low solubility. The international application WO201 3057518, from MW Encap, for example, discloses processes for the manufacture of suspensions comprising nanocrystals of one or more water-soluble or water-insoluble pharmaceutical or nutraceutical active ingredients with a particle size in the range of 0.01 to 10 microns. More specifically, the suspensions prepared by this process can be used to formulate pharmaceutical compositions, especially in liquid-filled capsules.

[0025] International patent application WO2017072774, from Solubest, discloses a pharmaceutical composition for transmucosal administration of an active lipophilic compound through the oral mucosa comprising a lipophilic active compound, a polymeric matrix formed by two or more water-soluble polymers and a fast-dissolving agent. At least one of the water-soluble polymers is an amphiphilic polymer and at least one is a hydrophilic polymer or an amphiphilic polymer with a hydrophobic-hydrophilic balance different from the first amphiphilic polymer. Furthermore, the polymeric matrix is ​​not cross-linked and there is no covalent interaction between the two or more polymers and between the polymers and the lipophilic active compound, which is intertwined with the aforementioned polymeric matrix. The said lipophilic active compound is selected from a group that includes anti-inflammatory agents. Examples are given in the descriptive report where the active ingredient is presented in nanocrystal form.

[0026] International patent application W00018374, from Elan Pharma International, discloses controlled-release nanoparticulate formulations comprising a nanoparticulate agent to be administered and a rate-control polymer that functions to prolong the release of the agent after administration. The claimed compositions release the agent after administration for a period ranging from about two to about twenty-four hours or more. It is mentioned in the descriptive report that the drug can be selected from anti-inflammatory, anthelmintic, antiarrhythmic, and antiasthmatic agents.

[0027] International patent application WO2018187728, from Maa Laboratories, discloses methods for preparing nanotherapeutic compounds and compositions comprising nanotherapeutic compounds. The nanotherapeutic compounds prepared according to the methods described are useful for the treatment of diseases, for example, cancer, in a subject in need. It is claimed that the therapeutic agent is selected from a group that includes anti-inflammatories.

[0028] International patent application WO2013132457, from the National Institute of Pharmaceutical Education & Research, discloses nanocrystalline solid dispersion compositions with discrete particles, wherein each discrete particle comprises crystals of at least one pharmaceutical active ingredient; veterinary active ingredient; nutraceutical active ingredient dispersed in a matrix of at least one crystallization inducer and / or coexisting with crystals of crystallization inducer, optionally together with pharmaceutically acceptable excipients. The present invention also encompasses a novel one-step process for generating nanocrystalline solid dispersions. The said active ingredient is selected from a group that includes non-steroidal anti-inflammatory agents.

[0029] Patent document CN108542886, from Liaocheng University, discloses a method for preparing celecoxib nanocrystals from the following components in weight percentage: 60-85 percent celecoxib, 10-30 percent a spatial stabilizer, and 1-10 percent an ionic stabilizer. According to the description, a suspension of celecoxib nanocrystals is prepared by a wet milling method, followed by lyophilization or spray drying to produce a solid powder.

[0030] Patent document BR 0210665, from Pharmacia Corporation, discloses a method for systemic administration of a hydrophobic substance to a mammal, comprising distributing the substance to the mammal's dermis, the substance having a molecular mass of 1000 daltons or less, which would include valdecoxib, celecoxib, or parecoxib.

[0031] Patent document CN 114917188, Shenyang Pharmaceutical University, discloses a long-acting celecoxib nanosuspension, which is prepared by mixing celecoxib, a stabilizer, a small molecule suspending agent, and water using a wet milling method. The celecoxib nanocrystal exhibits a rod-shaped morphology.

[0032] International patent application WO2009 / 114695, from Elan Pharma International, discloses a nanoparticulate angiogenesis inhibitor composition comprising particles of an angiogenesis inhibitor or a salt having an average particle size of less than 2000 nm; and associated with the surface thereof by at least one surface stabilizer, wherein the angiogenesis inhibitor may be celecoxib.

[0033] Patent document CN113402463, from Jiangnan University, discloses a preparation of microcrystalline celecoxib and a stabilizing agent, for treating osteoarthritis or rheumatoid arthritis. The preparation comprises dissolving the stabilizing agent in water and dissolving celecoxib in absolute ethyl alcohol.

[0034] Thus, it is observed that the preparation of celecoxib microcrystals or nanocrystals has not been aimed at obtaining a faster pharmacological action. Summary of the Invention

[0035] The present invention aims to provide stabilized celecoxib nanocrystals or its derivatives for faster pharmacological action than traditional formulations containing this active ingredient.

[0036] Celecoxib nanocrystals or their derivatives according to the present invention are stabilized with uncharged polymers, particularly of low viscosity, for a faster onset of action of this anti-inflammatory drug.

[0037] In other embodiments, the low-viscosity polymer may be based on polyvinyl alcohol, low molecular weight polyethylene glycol (PEG), methacrylic acid and methyl methacrylate copolymers, Copolymers of ethylene oxide and propylene oxide, polyacrylic acid (carbomers), hydroxypropylcellulose (HPC), hydroxypropylmethylcellulose (HPMC), methylcellulose (MC), carboxymethylcellulose (CMC), polyethylene glycol (PEG), polyvinylpyrrolidone (PVP) and their copolymers.

[0038] According to a particular object of the present invention, celecoxib nanocrystals or their derivatives are nanomized and stabilized with a low-viscosity polymer by a wet milling process, using a ball mill, through the collision of zirconium oxide (ZrO2) spheres.

[0039] Pharmaceutical compositions, uses, and treatment methods associated with celecoxib nanocrystals or their derivatives according to the present invention are also objects of the present invention. Brief Description of the Figures

[0040] In order to better define and clarify the content of this patent application, the following figures are presented:

[0041] [Fig. 1] Figure 1 shows the particle size distribution profile of the formulations obtained with low viscosity PVA (NCR-CLX-PVA_4-88), medium viscosity PVA (NCR-CLX-PVA-20-88) and high viscosity PVA (NCR-CLX-PVA_40-88).

[0042] [Fig. 2] Figure 2 shows the celecoxib nanosuspension (NCR-CLX-SUSP) obtained on the DeltaVital.

[0043] [Fig. 3] Figure 3 presents the results of the stability study of the CLX+PVA nanosuspension over 180 days, subjected to 4, 25 and 40°C, analyzed for diameter and polydispersity index.

[0044] [Fig. 4] Figure 4 shows the dissolution profile of the Celecoxib nanocrystal suspension (NCR-CLX-SUSP), evaluated in 900 ml of phosphate buffer pH 6.8, using apparatus 2, at a speed of 75 rpm and a test time of 60 minutes.

[0045] [Fig. 5] Figure 5 shows SEM images of dry formulation samples containing celecoxib, where A) Wide-field microscopy of the commercially available formulation, 2 mm scale bar, B) Microscopy of A) Wide-field microscopy of the NCR-CLX-LF formulation, 2 mm scale bar; B) Wide-field microscopy of the NCR-CLX-SDSE formulation, 500 µm scale bar; C) Detailed microscopy of the commercial formulation, 100 µm scale bar; D) Detailed microscopy of the NCR-CLX-LF formulation, 100 µm scale bar; E) Detailed microscopy of the NCR-CLX-SDSE formulation, 100 µm scale bar.

[0046] [Fig. 6] Figure 6 shows the laser diffraction of the formulations before drying (NCR-CLX-SUSP), after spray drying (NCR-CLX-SDSE), and after fluidized bed drying (NCR-CLX-LF).

[0047] [Fig. 7] Figure 7 shows mean celecoxib concentration values ​​in mg / mL, measured after 48 hours of contact under agitation in commercially available formulations (CLX-Line), after spray drying (NCR-CLX-SDSE) and after fluidized bed drying (NCR-CLX-LF).

[0048] [Fig. 8] Figure 8 shows the average percentage concentration values ​​of celecoxib released in pH 6.8 medium over 24 hours of contact with the medium, under agitation at 75 RPM in the standard formulations (CLX-FOXIS), after spray drying without excipient (NCR-CLX-SPRAY WITHOUT EXCIPIENT), after spray drying with excipient (NCR-CLX-SPRAY WITH EXCIPIENT) and after fluidized bed drying (NCR-CLX-FLUIDIZED BED).

[0049] [Fig. 9] Figure 9 shows the pharmacokinetic study in rats, presenting the plasma concentration of celecoxib in rats as a function of time after oral administration for 24 h (n=4), in detail the plasma concentration of celecoxib in the first two hours after administration of the formulations.

[0050] [Fig. 10] Figure 10 presents a panel with the pharmacokinetic results of commercial celecoxib formulations (FOXIS - Blue) and celecoxib nanocrystals obtained by spray drying, with the presence of drying excipients (NCR-CLX-SDCE - Red).

[0051] [Fig. 11] Figure 11 shows a panel with the analgesic efficacy assay results of celecoxib physical mixture (CLX-RM) and celecoxib nanocrystals (NCR-CLX), administered 1 hour and 15 minutes before the paw withdrawal time (PWL) assay performed after half an hour, 1 hour and 2 hours of carrageenan administration.

[0052] [Fig. 12] Figure 12 shows paw thickness after carrageenan injection, where paw thickness was checked after 30 minutes, 1, 2, 3, and 5 hours for the celecoxib nanocrystal suspension (NCR-CLX), physical mixture (RM), and control (vehicle) formulations. All formulations were administered 15 minutes before carrageenan injection. Data expressed as Mean±SEM, n=8. *p<0.05, **p<0.01, ***p<0.001 compared with the vehicle group using two-way ANOVA followed by Dunnett's multiple comparison test. Detailed Description of the Invention

[0053] Celecoxib is a selective COX-2 inhibitor with low water solubility and high permeability through biological membranes (Biopharmaceutical Classification System (BCS) class II - Dolenc, A., Kristi, J., Baumgartner, S., & Planinsek, O. (2009). Advantages of celecoxib nanosuspension formulation and transformation into tablets. International Journal of Pharmaceutics, 376(1-2), 204-212; Martinez, LM, Cruz-Angeles, J., Vázquez-Dávila, M., Martinez, E., Cabada, P., Navarrete-Bernal, C., & Cortez, F. (2022). Co-Crystals, or Polymorphs.

[0054] It was observed that nanomization of celecoxib promoted faster solubility and absorption, and increased drug efficacy.

[0055] The average particle size of celecoxib nanocrystal according to the present invention can vary between 10 and 1000 nm, preferably 200 nm to 503 nm.

[0056] The nanocrystals according to the present invention additionally exhibit the characteristics of (i) polydispersity index (PDI) ranging from 0.1 to 0.5, preferably 0.2, (ii) Zeta potential from -30 to +30 mV and (iii) maintenance of the crystallinity of the material after the comminution process, preferably maintaining the main XRD diffraction peaks and crystal shape by Electron Microscopy.

[0057] Thus, the present invention relates, in a first embodiment, to celecoxib nanocrystals or its derivatives, stabilized with polymers, preferably polyvinyl alcohol (PVA), particularly of low viscosity, for a faster onset of action of this anti-inflammatory drug. According to the present invention, low viscosity polymers are understood to be those having a viscosity of 1.0 - 21.9 mPa.s, particularly 3.4 - 4.6 mPa s, when in 40 g / L of water. Without imposing limitation, the PVA according to the preferred embodiment of the present invention is PVA 4-88.

[0058] The polymer features according to the present invention (1) a degree of hydrolysis (USP) ranging from 70 - 95%, preferably 85 - 89%, (2) a molecular weight ranging from 10,000 - 100,000 Da, preferably 13,000 - 23,000 Da, (3) a pH of 3 - 8, preferably 5.0 - 6.5, and (4) a linear or branched structure, preferably linear.

[0059] The nanocrystal according to the present invention can be in liquid form as a nanosuspension / nanodispersion or in solid form.

[0060] The term celecoxib derivatives covered by the present invention means any molecular forms of celecoxib according to the prior art.

[0061] The influence of PVA viscosity on the formation, interaction, and stabilization of nanocrystals obtained according to the present invention was observed. It can be inferred that as the viscosity of the PVA solution is reduced, the collisions of the zirconia oxide spheres are more effective, promoting a more effective nanomization of the crystals. Furthermore, Furthermore, because they have a shorter polymer chain, the polymer-nanocrystal interactions become more effective.

[0062] PVA is a polymer traditionally used for coating tablets and granulating powders, as well as increasing viscosity in liquid formulations and forming a matrix in semi-solid formulations. The use of PVA for spherical stabilization, located at the interface between nanocrystals and the aqueous medium (nanosuspension), of a nanocrystal formulation has not been described in the prior art.

[0063] Another important factor in choosing the formulation containing low-viscosity PVA was the absence of surfactants in its composition, as these can represent a risk for administration in biological systems through solubilization of the plasma membrane or formation of pores that destabilize the ionic balance of cells. Due to its chemical structure, low-viscosity PVA has a high degree of hydrolysis and does not have well-characterized hydrophilic and hydrophobic regions, thus having a reduced capacity to form micelles when compared to less hydrolyzed PVAs. Consequently, it has a reduced capacity to form pores in the plasma membrane of cells (Umemoto, Y., Ilchida, S., Yoshida, T., Shimada, K., Kojima, H., Takagi, A., Tanaka, S., Kashiwagura, Y., & Namiki, N. (2020). An effective polyvinyl alcohol for the solubilization of poorly water-soluble drugs in solid dispersion formulations. Journal of Drug Delivery Science and Technology, 55.).

[0064] In general, the state of the art teaches formulations containing nanocrystals stabilized by polymers and surfactants in order to maintain the system stable in liquid form and redispersible in solid form. The formulation that is the subject of the present invention fulfills both requirements without any presence of surfactants or filled polymers.

[0065] Anionic and cationic polymers tend to undergo ionization effects, which cause changes in the rate of dissolution and absorption along the gastrointestinal tract (e.g., carboxymethylcellulose (CMC) and acrylates). Few nonionic polymers, such as HPMC, PVP, and PVPVA, do not exhibit adjustable ratios of hydrogen bond donors and acceptors.

[0066] PVA has an adjustable density of hydrogen bond donor and hydrogen bond acceptor hydroxyl groups throughout its polymeric structure, yet it still has pH-independent solubility. In theory, this would allow it to function strongly as a solubility enhancer like anionic polymers, but it would also have the low variability of nonionic polymers.

[0067] The low-viscosity PVA used as a stabilizer according to the present invention is surfactant-free and pH-independent for use as a solubility enhancer, thus providing improved particle size, polydispersity index, and stability results for 6 months at 25°C.

[0068] The nanocrystal according to the present invention exhibits at least a 2-fold increase in the aqueous solubility of the active ingredient. Furthermore, it provides a rapid onset of anti-inflammatory and analgesic action observed at least 30 minutes after administration.

[0069] In another embodiment, the present invention is also related to the process of preparing celecoxib nanocrystals or its derivatives, which involves a top-down methodology using wet milling, particularly with a ball mill, through the collision of zirconium oxide (ZrO2) spheres. This preserves most of the active ingredient's crystallinity, reducing the formation of amorphous particles that may undergo structural alteration during storage. (Malamatari, Maria, et al. "Agglomerates of indomethacin nanoparticles: the role of poloxamers and matrix former in their resolution and aerosolization efficiency." International Journal of Pharmaceutics 495.1 (2015): 516-526; Colombo, M., Orthmann, S., Bellini, M. et al. Influence of Drug Brittleness, Nanomilling Time, and Freeze-Drying on the Crystallinity of Poorly Water-Soluble Drugs and Its Implications for Solubility Enhancement. AAPS PharmSciTech 18, 2437- 2445 (2017).).

[0070] In a particular embodiment, the active powder and a low-viscosity PVA solution in water were collided with ZrO2 spheres in a double centrifuge for approximately 4 hours, through rotational and translational movements, under refrigeration. This process was optimized through experimental design to obtain the optimal parameters; the input variables analyzed were CLX concentration (10% to 20%), PVA solution concentration (5% to 10%), and the quantity of Zirconia spheres (500 mg to 900 mg).

[0071] The optimal formulation selected according to the process of the present invention was that with about 15% celecoxib, about 5% to 10% PVA and about 500 to 900 mg of zirconia oxide maintained in a grinding process for about 240 minutes at about 1500 RPM.

[0072] The process according to the present invention may include a drying step that can be carried out by freeze-drying, vacuum evaporation / drying, air drying, atomization drying, spray drying, fluidized bed drying, convection drying (ovens), rotary evaporation or adsorption.

[0073] In another embodiment, the present invention relates to pharmaceutical compositions comprising celecoxib nanocrystals or derivatives thereof according to the present invention in a mixture with pharmaceutically acceptable excipients.

[0074] Suitable excipients according to the present invention may include, but are not limited to, excipients such as lactose and its variations, cellulose and its variations, maltitol, sucrose and its variations, sorbitol, maltose and its variations, starch and its variations, magnesium carbonate and its variations, magnesium oxide and its variations, dextrose and its variations, acrylates and their variations, alginates and their variations, dextrins and their variations, erythritol, fructose, talc and its variations, trehalose and its variations. variations, xylitol and its variations, glycine and its variations, carbonate, phosphate, silicate, calcium sulfate, alginic acid, alginate and its variations, cellulose and its variations, chitosan and its variations, colloidal silicon dioxide, starch and its variations, croscarmellose and its variations, crospovidone and its variations, sodium docusate, glycine, guar gum, xanthan gum and its variations, sodium lauryl sulfate and its variations, benzethonium chloride and its variations, benzalkonium chloride and its variations, cetylpyridine chloride and its variations, phospholipids and their variations, sodium docusate, hypromellose and its variations, poloxamer and its variations, polyoxyethylene and its variations, sorbitan esters, tricaprylin, vitamin E and its variations.Finally, binders such as povidone, alginate and its variations, chitosan, chlorpheniramine maleate, copovidone, phthalates and their variations, dibutyl sebacate, lactate and its variations, cellulose and its variations, gelatin and its variations, hypromellose and its variations, maleic acid and its variations, maltodextrin, dextrose and its variations, polyethylene glycol, polyethylene and its variations, acrylate and its variations, or mixtures thereof, are included.

[0075] In another embodiment, the present invention also relates to the use of celecoxib nanocrystals or their derivatives according to the present invention.

[0076] The uses and treatment methods associated with celecoxib nanocrystals or their derivatives according to the present invention are also objects of the present invention. Without imposing limitations, celecoxib nanocrystals or their derivatives according to the present invention are suitable for the treatment of pain relief, tenderness, swelling and stiffness caused by osteoarthritis, rheumatoid arthritis or juvenile rheumatoid arthritis in children 2 years of age and older. Examples

[0077] The examples shown here are intended to illustrate some of the numerous ways of carrying out the invention, without, however, limiting the scope. its purpose is to assist in proving the technical effect, in a direct and comparative way. Example 1 - Comparative formulations

[0078] Comparative formulations containing different proportions of celecoxib and stabilizers were developed. The responses evaluated were diameter and polydispersity index (Table 1).

[0079] These nanocrystals were prepared by a wet milling process through the collision of zirconium oxide (ZrO2) spheres. Briefly, the active powder and the PVA solution in water were collided with ZrO2 spheres in a double centrifuge for 4 hours, through rotational and translational movements, under refrigeration. The resulting nanosuspension was removed using a sieve with a 100 micrometer cutoff and deposited in a flask and kept under refrigeration.

[0080] [Table 1] Compositions of the nanocrystals. *active **Stabilizers (Soluplus - polyvinyl caprolactam - polyvinyl acetate grafted with polyethylene glycol copolymer; PVA 4-88 - Polyvinyl alcohol with an average viscosity of 4 mPas and 88% hydrolysis; PVA 26-88 - Polyvinyl alcohol with an average viscosity of 26 mPas and 88% hydrolysis; PVA 40-88 - Polyvinyl alcohol with an average viscosity of 40 mPas and 88% hydrolysis; Tween 80 - Polysorbate 80; Povacoat - Polyvinyl alcohol / Acrylic acid / Methyl methacrylate copolymer; Eudragit RS 30D - Ethyl acrylate, methyl methacrylate copolymer; PVP - Polyvinylpyrrolidone; Kollicoat MAE - Methacrylic acid / ethyl acrylate copolymer dispersion).** a Unstable for up to 1 month when stored at 25°C. NA - Not applicable, refers to formulations for which it was not possible to obtain a response, whether due to the gelation process, foam formation, or failure to form a suspension.

[0081] The results shown in Table 1 indicate that various types of polymers and surfactants were used to identify the most suitable one for developing a formulation with the highest possible active ingredient load, smallest particle size, and PDI, while remaining stable for the longest time.

[0082] As can be seen from comparative tests, low viscosity PVA, i.e., PVA 4-88, was the stabilizer that best met these characteristics, as it has an ideal viscosity of 3.4 - 4.6 mPa s when in 40 g / L of water.

[0083] The ideal viscosity has a strong impact on nanocrystal formation, as it allows for more effective collisions of the beads (grinding spheres) with the drug crystals, resulting in better and more homogeneous nanomization of the API (Active Pharmaceutical Ingredient), which can be seen through the particle size and PDI (Potential Dioxide Design) of the samples. The same can be observed when increasing the... The concentration of polymer in the medium causes the viscosity to increase.

[0084] Tests with PVAs of different viscosities showed their strong impact on the nanomization of crystals, as seen for the formulations containing PVA 20-88 (F19) and PVA 40-88 (F20). The particle size results showed that the formulations containing PVA with intermediate (20 mPas) and high (40 mPas) viscosity had a larger diameter and were more polydisperse. The three formulations obtained using PVA with the three different viscosities were the following: NCR-CLX-PVA_4-88 (low viscosity), NCR-CLX-PVA_20-88 (medium viscosity), and NCR-CLX-PVA_40-88 (high viscosity), and their particle size profile after 21 days of stability is shown in Figure 1.

[0085] Figure 1 shows the influence of PVA viscosity on the formation, interaction, and stabilization of the obtained nanocrystals. It can be inferred that as the viscosity of the PVA solution is reduced, the collisions of the zirconia oxide spheres become more effective, promoting a more effective nanomization of the crystals. Furthermore, due to their shorter polymer chain, the polymer-nanocrystal interactions become more effective. Example 2 - Characterization of NCRs

[0086] Particle size and zeta potential analyses of NCR-CLX-SUSP suspensions were performed.

[0087] To investigate the influence of PVA on the desired response (size, appearance, PDI) of the formulation, a Box-Behnken Design (BBD) with 3 factors at 3 levels was initially performed according to Table 1, in which the range of variation of each excipient was determined based on previously developed formulations. The experimental groups resulting from the combinations of the independent variables are shown in Table 2.

[0088] [Table 2] Variables and levels in the Box-Behnken method for the preparation of CLX nanocrystals. Celecoxib; ZIR: Zirconia spheres; PVA: Polyvinyl alcohol.

[0089] The hydrodynamic diameter, reported as Z-average, and polydispersity index (Pdl) of the nanosuspensions were determined by dynamic light scattering. The appearance was determined subjectively by visual analysis (Table 3).

[0090] [Table 3] Different proportions of compositions of NCR-CLX-SUSP suspensions tested in the BBD model with variations in the amount of celecoxib (CLX), polyvinyl alcohol (PVA) and zirconia oxide spheres (ZrO2).

[0091] Formulation 7 (Table 3) presented the most satisfactory results and, therefore, was selected to continue the experimental optimization of the process for obtaining nanodispersions in a double centrifuge by varying the process time, the number of rotations per minute, and the number of spheres by weight through a new BBD test (Table 4).

[0092] Based on the results obtained in the development of the formulations and the optimization of the process parameters, the selected formulation was that with 15% Celecoxib, 5% PVA and 900 mg of zirconia oxide maintained in a grinding process for 150 minutes at 1500 RPM (Table 5), an example of the nanosuspension can be seen in Figure 2.

[0093] [Table 4] Average hydrodynamic diameter (DH) and polydispersity index (PDI) of celecoxib nanocrystals in the test array of the response surface design of experiments for the variables Test time in minutes, Rotations per Minute (RPM) and number of beads in milligrams.

[0094] [Table 5] Nanosuspension composition and production process parameters. Example 3 - Stability

[0095] The optimized nanosuspension of celecoxib stabilized with PVA was evaluated for its stability at three different temperatures (4°C, 25°C and 40°C). The parameters analyzed were diameter and polydispersity index (Figure 3).

[0096] Figure 3 shows that the size of the nanocrystals and the homogeneity, expressed as polydispersity, remained constant during the 180 days of the study when stored at different temperatures (4, 25, and 40°C). This allows us to infer that this drug delivery system exhibits high stability. Example 4 - Dissolution profile

[0097] The dissolution profile of the CLX-NC nanosuspension was compared with the dissolution profiles of the physical mixture suspension of Celecoxib+PVA (PM). After particle size reduction, it was possible to quantify CLX in phosphate buffer pH 6.8. In the dissolution profile of the nanocrystals, we observed a release of up to 27.78% after 60 minutes of testing, a higher value when compared to the physical mixture (PM), which had a release of less than 2% of the drug after 60 min (Figure 4). Example 5 - Drying of NCR-CLX-SUSP nanosuspensions

[0098] The nanocrystals obtained through the wet milling process were dried using two different techniques. The first was by spray drying the suspension in a fluidized bed (NCR-CLX-LF) with excipients from the commercial formulation (lactose monohydrate, croscarmellose sodium, sodium lauryl sulfate, and povidone). The second drying technique used was spray drying, which consists of drying the nanocrystal suspension to obtain a dry powder. In this second technique, two formulations were tested: the first in which no excipients were used for the drying process (NCR-CLX-SDSE), and the second in which mannitol and sodium lauryl sulfate were used as excipients during drying (NCR-CLX-SDCE).

[0099] Besides the methods used (spray dryer and fluidized bed), other techniques are possible for obtaining a powder formulation from... The methods of drying a liquid include convection drying (ovens), rotary evaporation, freeze-drying, and liquid adsorption. Example 6 - Characterization of the obtained powders

[0100] The size and morphology of the NCR-CLX powder particles were determined by different techniques, presented here by Scanning Electron Microscopy (SEM) in Figure 4 (AF) and Laser Diffraction (LD). The first was performed on the spray-dried powder, while the second technique was determined in aqueous medium using the refractive index of the outermost layer (PVA), as shown in Figure 6 and Table 6. Solubility was determined by dispersing excess celecoxib in phosphate buffer solution pH 6.8 for 48 hours in an orbital shaker followed by filtration through a 0.45 µm PTFE filter and quantification by UV-Vis HPLC (Figure 7). The dissolution of the samples was carried out in Apparatus II dissolution apparatus in phosphate buffer pH 6.8 with 0.1% sodium lauryl sulfate, with collections at times 15, 30, 60, 120, 240, 480 and 1440 minutes. The aliquots were filtered through a 0.45 µm PTFE filter and quantified by HPLC UV Vis (Figure 8).

[0101] [Table 6] Average particle size and dispersibility (Span) values ​​obtained by laser diffraction of the formulations before drying (NCR-CLX-SUSP), after spray drying (NCR-CLX-SDSE) and after fluidized bed drying (NCR-CLX-LF).

[0102] The powder characterization results showed that the spray-dried nanoparticle clusters had superior solubility, with greater and faster dissolution than the other groups analyzed. Therefore, they were chosen to proceed with in vivo pharmacokinetic assays in dogs. Example 7 - Pharmacokinetic study in rats

[0103] The oral bioavailability of celecoxib nanosuspension was evaluated in rats. The comparative groups administered to the animals were: physical mixture (non-nanomized CLX + PVA) and CLX nanocrystals.

[0104] The objective of this study was to evaluate the increase in the bioavailability of the active ingredient offered by the technology. After administration of the samples by gavage, the plasma of the animals was collected (0.25; 0.5; 1; 2, 4, 8, 12 and 24h) and the mean plasma concentration of celecoxib quantified versus time is shown in Figure 9.

[0105] Pharmacokinetic parameters such as Cmax, Tmax, T1 / 2, AUC1ast, MRTO1ast, and bioavailability are presented in Table 7. Oral absorption of CLX in the nanosuspension increased compared to the physical mixture sample (Figure 9). The Cmax value in the nanosuspension was 4.868 ± 0.807 pg / mL and in the physical mixture it was 2.697 ± 0.679 pg / mL, demonstrating the increase in bioavailability and increased onset offered by the technology.

[0106] [Table 7] Pharmacokinetic parameters of Celecoxib physical mixture (MF-CLX) and Nanocrystal Suspension (NCR-CLX-SUSP) after oral administration in rats (n=4, mean ± SD). Example 8 - Pharmacokinetic study in dogs

[0107] The oral bioavailability of nanomized and spray-dried CLX was evaluated in dogs. Comparative groups were administered to the animals. The samples consisted of: a standard product (Foxis 200 mg) and CLX nanocrystals dried using a spray dryer.

[0108] The objective of this study was to evaluate the increase in the bioavailability of the active ingredient offered by the technology after spray drying. After sample administration, animal plasma was collected (0.25, 0.5, 1, 2, 4, 8, 12, and 24 h) and the mean plasma concentration of celecoxib was quantified versus time, as shown in Figure 10. Pharmacokinetic parameters such as Cmax, Tmax, T1 / 2, AUC1ast, MRTO1ast, and bioavailability are presented in Table 8.

[0109] [Table 8] Pharmacokinetic parameters of celecoxib after oral administration in rats (n=4, mean ± SD).

[0110] Pharmacokinetic studies in Beagle dogs showed that the capsule formulation with spray-dried celecoxib nanocrystal powder with drying excipients (NCR-CLX-SDCE) had superior and significantly higher absorption and bioavailability than the commercial celecoxib formulation (FOXIS). When comparing Cmax, we observed a surprising 2.8-fold increase offered by the technology (4713 ng / mL versus 1685 ng / mL) (Table 8 and Figure 10). Example 9 - Efficacy Study in Rats

[0111] After verifying the higher oral bioavailability of celecoxib in both rats and dogs, an efficacy assay was performed to observe a possible increase in the analgesic and anti-inflammatory efficacy of celecoxib when nanosized (NCR-CLX). The model used was the induction of inflammation by means of carrageenan, and paw thickness (swelling) and paw withdrawal time after thermal stimulation (thermal hyperalgesia) were evaluated.

[0112] Formulations containing celecoxib nanocrystals (NCR-CLX) and a physical mixture of celecoxib and PVA (CLX-RM) were tested. The formulations were administered 1 hour or 15 minutes (0.25 h) before inflammatory stimulation with carrageenan. Figure 11 shows the results obtained after thermal stimulation, where the time to remove the paw from a heated plate was measured after 0.5, 1, and 2 hours.

[0113] In parallel with the thermal stimulus test, paw swelling was also measured by evaluating its thickness over 5 hours, verifying the anti-inflammatory power of each of the formulations (Figure 12).

[0114] The NCR-CLX formulation showed greater anti-inflammatory capacity, significantly reducing paw swelling at all points when compared to the physical mixture (CLX-RM) and control (vehicle) groups. This assay was performed only on the groups that received the formulations 15 minutes before the carrageenan injection.

[0115] The NCR-CLX formulation showed a more favorable onset for paw removal from a heated plate (sensitization test), meaning the animal kept its paw on the plate for a longer time (less sensitivity / pain), with a statistically significant difference exclusively for the NCR-CLX group after 0.5 hours of carrageenan administration. This demonstrates a superior, surprising, and rapid onset of action.

[0116] The results of the comparative tests showed that the formulations were obtained through a reproducible scientific methodology, demonstrating pharmaceutical advantages over products currently on the market.

[0117] Reducing the size of celecoxib crystals increased the dissolution of the active ingredient by an average of 25 times at physiological pH (6.8) and 28 times the Cmax in a pharmacokinetic study in dogs.

[0118] Furthermore, it was possible to dry the nanosuspensions using fluidized bed drying and spray drying techniques.

[0119] The developed CLX nanosuspension formulation showed a faster onset of action than the physical mixture of celecoxib (active ingredient + PVA) when administered at the same concentration.

[0120] CLX nanocrystals increased the oral bioavailability, Cmax, and Tmax of CLX when compared to the currently available product on the market, Foxis, demonstrating superior efficacy.

[0121] Those skilled in the art will appreciate the knowledge presented here, which allows them to reproduce the invention in the forms presented and in other variations covered by the appended claims.

Claims

Claims

1. CELECOXIB NANOCRYSTAL characterized by being stabilized with low viscosity polymers.

2. NANOCRYSTAL, according to claim 1, characterized in that the low viscosity varies between 1.0 - 21.9 mPa s when in 40 g / L of water.

3. NANOCRYSTAL, according to claim 2, characterized in that the low viscosity varies between 3.4 and 4.6 mPa s when in 40 g / L of water.

4. NANOCRYSTAL, according to claim 1, characterized in that the polymer has (1) a degree of hydrolysis (USP) ranging from 70 to 95%, (2) a molecular weight ranging from 10,000 to 100,000 Da, (3) a pH of 3 to 8, and (4) a linear or branched structure.

5. NANOCRYSTAL, according to claim 4, characterized in that the polymer has (1) a degree of hydrolysis (USP) ranging from 85 to 89%, (2) a molecular weight ranging from 13,000 to 23,000 Da, (3) a pH of 4% in H2O of 5.0 to 6.5 and (4) a linear structure.

6. NANOCRYSTAL, according to any one of claims 1 to 5, characterized in that the low viscosity polymer is based on polyvinyl alcohol (PVA), low molecular weight polyethylene glycol (PEG), methacrylic acid and methyl methacrylate copolymers, ethylene oxide and propylene oxide copolymers, polyacrylic acid (carbomers), hydroxypropylcellulose (HPC), hydroxypropylmethylcellulose (HPMC), methylcellulose (MC), carboxymethylcellulose (CMC), polyethylene glycol (PEG), polyvinylpyrrolidone (PVP) and their copolymers.

7. NANOCRYSTAL according to claim 1, characterized by being in liquid form as a nanosuspension / nanodispersion or in solid form.

8. NANOCRYSTAL, according to claim 1, characterized in that the particle size of the celecoxib nanocrystal varies between 10 and 1000 nm.

9. NANOCRYSTAL, according to claim 8, characterized in that the average particle size of the celecoxib nanocrystal varies between 100 and 500 nm.

10. NANOCRYSTAL, according to claim 1, characterized in that (i) the polydispersity index (PDI) ranges from 0.1 to 0.5, (ii) the Zeta potential is from -30 to +30 mV and (iii) the crystallinity of the material is maintained after the comminution process.

11. NANOCRYSTAL, according to claim 1, characterized in that (i) the polydispersity index (PDI) varies between 0.1 and 0.3 and (iii) the main XRD diffraction peaks and the crystal shape by Electron Microscopy are maintained.

12. NANOCRYSTAL, according to claim 1, characterized in that it includes celecoxib derivatives.

13. PROCESS FOR PRODUCING CELECOXIB NANOCRYSTAL, as defined in any one of claims 1 to 12, characterized by using top-down methods.

14. PROCESS, according to claim 13, characterized in that the top-down method includes wet or dry grinding with a ball mill, high-energy grinding, high-pressure homogenization and similar comminution processes.

15. PROCESS, according to claim 14, characterized in that the high-pressure homogenizer utilizes a pressure range of 100 to 2000 bar and a number of 1 to 20 cycles.

16. PROCESS, according to claim 14, characterized in that the ball mill uses a ball filling of 75% to 90%, an agitator speed of 2000 rpm to 3600 rpm and a pump flow rate of 30 to 185 rpm.

17. PROCESS, according to claim 13, characterized in that to obtain a nanocrystal powder, freeze-drying, vacuum evaporation / drying, surface drying (air drying), atomization drying (spray drying), fluidized bed drying, convection drying (ovens), rotary evaporation, solid fragmentation techniques and liquid adsorption techniques are used.

18. PHARMACEUTICAL COMPOSITION characterized in that it comprises celecoxib nanocrystal, as defined in any one of claims 1 to 12, and pharmaceutically acceptable excipients.

19. COMPOSITION, according to claim 18, characterized in that the pharmaceutically acceptable excipients are selected from the group comprising: lactose and its variations, cellulose and its variations, maltitol, sucrose and its variations, sorbitol, maltose and its variations, starch and its variations, magnesium carbonate and its variations, magnesium oxide and its variations, dextrose and its variations, acrylates and their variations, alginates and their variations, dextrins and their variations, erythritol, fructose, talc and its variations, trehalose and its variations, xylitol and its variations, glycine and its variations, calcium carbonate, phosphate, silicate, calcium sulfate, alginic acid, alginate and its variations, cellulose and its variations, chitosan and its variations, colloidal silicon dioxide, starch and its variations, croscarmellose and its variations, crospovidone and its variations, sodium docusate, glycine, guar gum, xanthan gum and their variations,Sodium lauryl sulfate and its variations, benzethonium chloride and its variations, benzalkonium chloride and its variations, cetylpyridine chloride and its variations, phospholipids and their variations, sodium docusate, hypromellose and its variations, poloxamer and its variations, polyoxyethylene and its variations, sorbitan esters, tricaprylin, Vitamin E and its variations; and the binders shall be selected from the group comprising povidone, alginate and its variations, chitosan, chlorpheniramine maleate, copovidone, phthalates and their variations, dibutyl sebacate, lactate and its variations, cellulose and its variations, gelatin and its variations, hypromellose and its variations, maleic acid and its variations, maltodextrin, dextrose and its variations, polyethylene glycol, polyethylene and its variations, acrylate and its variations, mannitol, or mixtures thereof.

20. USE OF A CELECOXIB NANOCRYSTAL, as defined in any one of claims 1 to 12, for the treatment of conditions related to pain and inflammation.

21. TREATMENT METHOD FOR RELIEVING CONDITIONS RELATED TO PAIN AND INFLAMMATION, characterized by comprising a step of providing the patient in need with celecoxib nanocrystals as defined in any one of claims 1 to 12.

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