A nanoparticle formulation containing boron nitride

Abstract

The present invention relates to a nanoparticle adjuvant formulation containing boron nitride, provided in the form of a nanosuspension prepared by a nanoprecipitation technique, and suitable for use in the healthcare field to provide bacterial inactivation as well as vaccine adjuvant activity.

Description

[0001] A NANOPARTICLE FORMULATION CONTAINING BORON NITRIDE

[0002] Technical Field

[0003] The present invention relates to a nanoparticle adjuvant formulation containing boron nitride, provided in the form of a nanosuspension prepared by a nanoprecipitation technique, and suitable for use in the healthcare field to provide bacterial inactivation as well as vaccine adjuvant activity.

[0004] Background of the Invention

[0005] Among the adjuvants currently used in vaccines both domestically and worldwide, aluminum salts, montanide, saponins, proteins, and virus-like particles are commonly employed. These vaccines have remained largely unchanged over time. However, as one of the most preferred vaccine adjuvant candidates, aluminum salts are associated with risks arising from their extensive use, including aluminum toxicity and high-level binding to transferrin in the blood, thereby reducing iron-binding capacity. Aluminum salts may also inhibit magnesium transport in cells and disrupt calcium metabolism, leading to oxidative stress. Moreover, when present in high amounts, aluminum may cross the blood-brain barrier and accumulate in the brain. Disruption of calcium metabolism may further result in transfer to bone tissue. In addition, aluminum salts may cross the placental barrier and reach the fetus and embryo during pregnancy, potentially causing toxic effects and pronounced neurotoxicity. Studies have further shown that aluminum salts stimulate only humoral immunity and do not elicit a cellular immune response, thereby necessitating their combination with other substances.

[0006] A review of the prior art further indicates that vaccines currently in use are associated with substantial costs. In addition to said adjuvants, vaccine formulations typically require the inclusion of at least one (in some cases, at least two) chemical agent for inactivation. Moreover, certain vaccines are known to contain at least two vaccine adjuvants. In the case of multi-dose formulations, the inclusion of a preservative (such as phenol or thiomersal) is also required. Collectively, these factors contribute significantly to overall vaccine costs. Furthermore, vaccines currently in use may be manufactured domestically within thenational borders (in Turkiye); however, the adjuvants used in such vaccines are imported. In contrast to existing approaches, the nanoparticle adjuvant formulation containing boron nitride disclosed herein is capable of inhibiting bacteria while also stimulating the immune system within a single formulation, thereby providing a protective effect against a disease.

[0007] Description of the Invention

[0008] In the present description, the nanoparticle adjuvant formulation containing boron nitride is provided solely for facilitating a clearer understanding of the subject matter, without any intention to impose a limiting effect.

[0009] The present invention relates to a nanoparticle adjuvant formulation containing boron nitride, which satisfies the aforementioned requirements, eliminates existing disadvantages, and provides additional advantages.

[0010] The primary object of the invention is to provide an inhibitory effect while preserving antigenic determinants and to generate an immune response in the healthcare field.

[0011] The invention provides a nanoparticle adjuvant formulation containing boron nitride, which is provided in the form of a nanosuspension using a nanoprecipitation technique and is suitable for use in the healthcare field to provide bacterial inactivation, as well as vaccine adjuvant activity.

[0012] Most preferably, the invention provides a nanoparticle adjuvant formulation containing boron nitride, which provides both bacterial inhibition and vaccine adjuvant activity.

[0013] More preferably, the invention provides a nanoparticle adjuvant formulation containing boron nitride, wherein boron derivatives increase cytokine stimulation by enabling mononuclear cell proliferation in the blood.

[0014] More preferably, the invention provides a nanoparticle adjuvant formulation containing boron nitride, wherein boron nitride, as one of the underground resources within the national borders (in Turkiye), is converted for the first time into a form applicable to worldwide use as a vaccine adjuvant.More preferably, the invention provides a nanoparticle adjuvant formulation containing boron nitride, wherein boron nitride offers cost and raw material advantages due to its abundant availability within the country’s underground boron resources.

[0015] More preferably, the invention provides a nanoparticle adjuvant formulation containing boron nitride.

[0016] The components included in the invention are as follows:

[0017] a) Boron nitride: The primary component of the adjuvant formulation, which is responsible for bacterial inactivation and immune stimulation,

[0018] b) Captex 355: A surfactant used to convert boron nitride into nanoparticle form in the present adjuvant formulation,

[0019] c) Gelucire 48 / 16: A surfactant used to convert boron nitride into nanoparticle form in the present adjuvant formulation,

[0020] d) Acetone: An organic solvent used to convert boron nitride into nanoparticle form in the present adjuvant formulation, applied during the preparation process and subsequently removed from the medium, without forming part of the formulation, e) Ethanol: An organic solvent used to convert boron nitride into nanoparticle form in the present adjuvant formulation, applied during the preparation process and subsequently removed from the medium, without forming part of the formulation, f) Dimethyl sulfoxide: An organic solvent used to convert boron nitride into nanoparticle form in the present adjuvant formulation, applied during the preparation process and subsequently removed from the medium, without forming part of the formulation,

[0021] g) Ultrapure water: An essential component used at various stages of the adjuvant formulation.

[0022] A production method of the invention is as follows:

[0023] 1. Weighing: All components are weighed using a precision balance with a sensitivity of 1 mg.

[0024] 2. Mixing: The components in the vaccine formulation are mixed at 750 rpm for 30 minutes and dissolved.

[0025] 3. Sonication: Sonication is performed at 60% power for 5 minutes at a frequency of 2 cycles per second, while mixing is maintained at 750 rpm throughout the sonication process.4. Evaporation: The acetone-ethanol mixture is evaporated under vacuum at 45°C for 5 minutes using an evaporator.

[0026] 5. Ultracentrifugation: Centrifugation is carried out at 12,500 rpm for 30 minutes using a cooled ultracentrifuge at 15°C, thereby removing residual solvents and dimethyl sulfoxide (DMSO).

[0027] 6. Sonication: Another sonication step is performed at 60% power for 5 minutes at 2 cycles per second, while mixing is maintained at 750 rpm throughout the sonication process.

[0028] 7. UV Sterilization: The resulting nanoparticle formulations are sterilized under UV at a wavelength of 260 nm for a duration of 2 hours.

[0029] 8. Bacterial Inoculation: For sterility control, samples taken from the formulation are inoculated into liquid Thioglycollate culture medium and incubated at 37°C for 14 days under aerobic, anaerobic, and microaerobic conditions.

[0030] The mixing process may be carried out at 500-1000 rpm for 15-60 minutes, preferably at 750 rpm for 30 minutes. The sonication process is required to be performed at a power level of 25-75% for 1-15 minutes and at 1-4 cycles; preferably at 60% power for 5 minutes and at 2 cycles. Evaporation process is required to be performed under vacuum ata temperature of 30-75°C for 1-15 minutes, preferably at 45°C for 5 minutes. Ultracentrifugation may be carried out at 10,000-15,000 rpm for 10-60 minutes at a temperatures of 5-25°C, preferably at 12,500 rpm for 30 minutes at 15°C.

[0031] Captex 355 employed in the formulation may be replaced with at least one surfactant selected from the group consisting of: Compritol 888 ATO, Gelot 64, isopropyl myristate, sodium lauryl sulfate, Poloxamer 188, Tweens (such as 20, 40, 60, 80, 85), Spans (such as 20, 40, 60, 80, 85), lecithin, Gelucire® varieties, Caprylol 90, Lauroglycol 90, Labrasol®, Labrafil® varieties, Captex® varieties, Brij® 72, Capmul® varieties, Poloxamer 407, or a combination thereof.

[0032] Gelucire 48 / 16 employed in the formulation may be replaced with at least one surfactant selected from the group consisting of Compritol 888 ATO, Gelot 64, isopropyl myristate, sodium lauryl sulfate, Poloxamer 188, Tweens (such as 20, 40, 60, 80, 85), Spans (such as 20, 40, 60, 80, 85), lecithin, Gelucire® varieties, Caprylol 90, Lauroglycol 90, Labrasol®, Labrafil® varieties, Captex® varieties, Brij® 72, Capmul® varieties, Poloxamer 407, or a combination thereof.Acetone employed in the formulation may be replaced with other organic solvents such as ethanol, acetonitrile, dichloromethane, ethyl acetate, or methanol.

[0033] Ethanol employed in the formulation may be replaced with other organic solvents such as acetone, acetonitrile, dichloromethane, ethyl acetate, or methanol.

[0034] Lactic acid employed in the formulation may be replaced with acetic acid.

Claims

CLAIMS1. A nanoparticle adjuvant formulation containing boron nitride, provided in the form of a nanosuspension prepared by a nanoprecipitation technique, characterized by comprising boron nitride, Captex 355, Gelucire 48 / 16, acetone, ethanol, dimethyl sulfoxide, and ultrapure water.

2. A formulation according to claim 1 , characterized in that the formulation comprises boron nitride at an amount of 0.1-25% (w / w), Captex 355 at an amount of 0.1-25% (w / w), Gelucire 48 / 16 at an amount of 0.1-25% (w / w), and ultrapure water at an amount of 1-95% (w / w).

3. A formulation according to claim 1 , characterized in that:• Captex 355 employed in the formulation can be replaced with at least one surfactant selected from the group consisting of: Compritol 888 ATO, Gelot 64, isopropyl myristate, sodium lauryl sulfate, Poloxamer 188, Tweens (such as 20, 40, 60, 80, 85), Spans (such as 20, 40, 60, 80, 85), lecithin, Gelucire® varieties, Caprylol 90, Lauroglycol 90, Labrasol®, Labrafil® varieties, Captex® varieties, Brij® 72, Capmul® varieties, Poloxamer 407, or a combination thereof.• Gelucire 48 / 16 employed in the formulation can be replaced with at least one surfactant selected from the group consisting of Compritol 888 ATO, Gelot 64, isopropyl myristate, sodium lauryl sulfate, Poloxamer 188, Tweens (such as 20, 40, 60, 80, 85), Spans (such as 20, 40, 60, 80, 85), lecithin, Gelucire® varieties, Caprylol 90, Lauroglycol 90, Labrasol®, Labrafil® varieties, Captex® varieties, Brij® 72, Capmul® varieties, Poloxamer 407, or a combination thereof.• Acetone employed in the formulation can be replaced with other organic solvents such as ethanol, acetonitrile, dichloromethane, ethyl acetate, or methanol.