TW202142259A - Use of nanoscale silicate platelet (nsp) for interrupting virus infection pathway - Google Patents

Abstract

The instant invention deals with the uses of nanoscale silicate platelets (NSP) for interrupting virus infection pathway. The high surface area and intensive negative charges on surface renders NSP physical adsorption and chemical charge exchanging neutralization, consequently reducing the virus infection.

Description

Methods of resisting viral infections with nano-silicon wafers

The present invention relates to a method for resisting viral infections with nano-silicon wafers, especially viruses with positive charges on the surface.

The basic structure of a virus is a protein shell coated with ribonucleic acid (RNA). Because the virus has the characteristics of rapid mutation and spread, it can be said to be the greatest enemy of mankind. Take the new type of coronavirus that causes COVID-19 as an example. Its basic structure includes internal ribonucleic acid (RNA) and external protein. Among the external proteins, the HIV-1 Gag protein is positively charged after dissociation and can bind to negatively charged host cell receptors and cause infection.

The literature has proposed the use of sodium lauryl sulfonate (SLS) to prevent HIV-1 infection, see: Piret J.; Désormeaux, A. & Bergeron, MG (2002). "Sodium lauryl sulfate, a microbicide effective against Enveloped and nonenveloped viruses". Curr. Drug Targets. 3(1): 17-30, and Piret J.; Lamontagne, J.; Bestman-Smith, J.; Roy, S.; Gourde, P.; Désormeaux, A .; Omar, RF; Juhász, J. & Bergeron, MG(2000). "In vitro and in vivo evaluations of sodium lauryl sulfate and dextran sulfate as microbicides against herpes simplex and human immunodeficiency viruses". J.Clin.Microbiol.38 (1): 110-119.

For positively charged viruses, there are also literatures suggesting that the viruses can be combined with negatively charged sodium dodecyl sulfonate (SDS) and nano-silicon wafers. (nanoscale silicate platelets, NSP) contact, and produce electrical neutralization, by blocking the route of infection, to achieve the purpose of epidemic prevention. See: Liang JJ, Wei JC, Lee YL, Hsu SH, Lin JJ, Lin YL. "Surfactant-Modified Nanoclay Exhibits an Antiviral Activity with High Potency and Broad Spectrum". J Virol. 2014 Apr; 88(8): 4218- 28. However, the document believes that the use of nano-silicon wafers (NSP aqueous solution, 0.001 wt%) alone cannot effectively prevent the Japanese encephalitis virus from multiplying.

It is known that nano-silicon wafer (NSP) is a kind of silicate with sheet-like geometry, which has a high surface area and a high density of negative charges. Figure 1 is a schematic diagram of the structure of a nano-silicon wafer, where (≡Si-O) is the main network structure, (≡SiOH) (siloxanol) has a hydrophilic hydrogen bond, and (≡SiO ^- Na ⁺ ) is anion/yang Or negative/positive ion pair. The composition of the latter two is conducive to hydrophilicity, so that the nano-silicon wafer can be dispersed and floated in the water, and has the surface energy to adsorb viruses. Nano silicon wafers can be manufactured from silicate clay, so the cost is low. In addition, nano silicon wafers can form a uniformly dispersed silicon wafer liquid in various concentrations in water, which increases the convenience of transportation and use. The most important thing is that nano-silicon wafers are not biologically toxic, and their cytotoxicity to humans is approximately equal to that of table salt (rat acute toxicity test LD ₅₀ > 5700 mg/kg). In other words, nano-silicon wafers still have high potential and R&D value in resisting viral infections. Therefore, the inventor further proposed the method of the present invention based on the characteristics of the nanosilicon wafer and the virus.

The purpose of the present invention is to provide a method for resisting viral infections with nano-silicon wafers, which includes the following steps:

(i) Provide an aqueous solution of nano-silicon wafers, the concentration of the nano-silicon wafers is 0.01wt%~10.0wt%, the nano-silicon wafers are completely delaminated silicic acid clay, and the wafer diameter ratio is (50~300)× (50~300)×1nm ³ , the surface area is 300m ² /g~800m ² /g, the surface structure includes ≡SiOH and ≡SiO ^- Na ⁺ , the cation exchange capacity is 1.0mequiv/g~1.5mequiv/g; and

(ii) Bring the nanosilicon wafer into contact with the charged virus, and absorb the virus by one or more of van der Waals force, hydrogen bond, and neutralization of electricity, so as to interrupt the free movement of the virus. Blocking the route of infection may even cause the virus to lose its activity and achieve the purpose of epidemic prevention.

As shown in Figure 1, there are three forces on the surface of each piece of NSP at the same time: (1) The flake geometry produces a very high surface area, and the following van der Waals force can adsorb any particles of nanometer size. Including viruses. (2) The hydrophilicity of ≡SiOH, which adsorbs viruses through hydrogen-bonded water media. (3) Ion exchange chemical reaction, similar to acid-base neutralization reaction, negative ion NSP can forcibly neutralize the positive valence structure of the virus, that is, the N ⁺ quaternary amino group of the protein. Therefore, NSP has high mobility and activity in water, and can capture, adsorb and neutralize viruses, physically making them lose their original infectivity. In other words, NSP can block the infection mechanism or probability of any virus entering the host cell, and it is even more so for "normal" viruses.

In addition, the anti-virus mechanism of nano-silicon wafers (NSP) depends on its own dispersion in water. In comparison, clay with poor dispersion is more prone to agglomerate state, the surface area cannot be expressed, the affinity with the virus (that is, Van der Waals force and ionic bond) is reduced, and it cannot exhibit nano-microscopic effects. Compared with layered clay, the dispersibility of nano-silicon flakes in water is significantly improved. NSP surface with a large amount of positive charge (Na ⁺⁾ and negative charges (SiO ^-), with bis characteristics and maintain electrical neutrality of balance, so that it can react with anionic / cationic dispersing agent or intercalant respectively, help For the dispersibility of NSP in water. The particle size of NSP and the raw clay montmorillonite (MMT) in the aqueous solution was analyzed with a dynamic light scattering (DLS). The results are shown in Table 1. The particles of NSP have become finer and have improved dispersibility.

In the method of the present invention, the concentration of the nano-silicon wafer is preferably 0.01 wt% to 0.1 wt%, more preferably 0.05 wt% to 0.1 wt%. The aqueous solution of nanosilicon chips can still include an anionic surfactant, such as sodium dodecyl/lauryl sulfonate (SDS/SLS), sodium stearate (sodium octadecanoate), dextran sulfate (dextran sulfate sodium, DSS), C12-14 alkyl ethoxy sulfate ((C12-14)alkyl ethoxy sulfate), C10-16 alkylbenzenesulfonic acid ((C10-C16)alkylbenzenesulfonic acid), ten Sodium dodecylbenzene sulphonate (SDBS), sodium xylene sulfonate (sodium xylene sulfonate), sodium toluene sulfonate (sodium toluene sulfonate), sodium cumene sulfonate (sodium cumene sulfonate), alkylbenzene Compounds such as calcium alkyl benzene sulfonate and sodium bis-naphthalene sulfonate, among which sodium dodecyl sulfonate (SDS) is preferred. The weight ratio of NSP to anionic surfactant is 10/90~90/10, preferably 10/90~70/30.

In order to promote the uniform dispersion of nano-silicon wafers in water, the aqueous solution of nano-silicon wafers can still include a non-ionic surfactant, such as Triton ^® X-100 (TEDIA, octylphenol polyethoxylate), SINOPOL ^® series (China-Japan Synthetic Chemical Company, polyethylene glycol ether, polyoxyethylene ether), etc.

The nano-silicon wafer aqueous solution can be directly or in the form of spray with the charged Virus contact, or make the nano-silicon wafer aqueous solution first adhere to the surface of an object, or evenly dispersed in the cytoplasm, to catch or interrupt the virus invasion and contact. By one or more of van der Waals force, hydrogen bond and electricity price neutralization, the activity of bacteria or viruses can be removed or reduced to achieve the function of epidemic prevention.

Figure 1 is a schematic diagram of the structure of the nano-silicon wafer.

Figure 2 shows the experimental results of the nanosilicon wafer and sodium dodecyl sulfonate in Example 1.

Figure 3 shows the experimental results of nanosilicon wafers and sodium lauryl sulfonate in Example 2.

The materials used in the preferred embodiments and comparative examples of the present invention include:

1. Nanoscale silicate platelets (NSP): Silicate clay, such as bentonite or montmorillonite, is a sheet-like structure formed by complete delamination. The geometric appearance has a high sheet diameter ratio of about (50~300)×(50~300)×1nm ³ , and the surface area can be as high as 720m ² /g. The surface ion density is about 18,000 ions/sheet, and the cation exchange capacity is about 1.2 mequiv/g. The manufacturing method of the nano-silicon wafer can refer to the methods of US Patent No. 7,022,299B2, No. 7,094,815B2, No. 7,125,916B2, and Announcement No. 2005-0239943-A1, and Application No. 11/464,495. The following is one of the methods, including the following steps:

(a). Disperse montmorillonite (10g) in hot water (1L) at 80°C and stir vigorously for 4 hours to make the aqueous solution form a stable and uniform dispersion of earthy color.

(b). Take p-cresol (27.2g) and D2000 (757.6g) at a molar ratio of 2:3, reflux in toluene at 90°C for 3 hours, then add formaldehyde solution (37wt%, 61.4g), and increase the temperature React at 130°C for 5 hours, and stop the reaction after forming a gel. The resulting product is Amine-termination Mannich Oligomer (AMO). GPC analysis showed that the three peak positions were Mw=3,142, 6,221 and 9,246; the amine titration results were: primary amine=0.4meq/g, secondary amine=0.56meq/g, no tertiary amine. AMO was dissolved in water (575g) and mixed with concentrated hydrochloric acid (35wt%, 36g) at 80°C for 30 minutes to form acidified AMO. Pour the acidified AMO into the above Na ⁺ -MMT dispersion and stir vigorously at 80°C for 5 hours to complete the intercalation reaction and obtain an AMO/Clay mixed solution.

(c). Add buffered aqueous solutions of different pH values to the AMO/Clay mixed solution. At this time, the AMO/Clay mixed solution will form a light yellow, emulsified viscous liquid.

(d). Add ethanol (7.5L) to the viscous liquid and filter it, then add ethanol (10L) to the filtered solid, stir evenly, add the equivalent of NaOH (9.2g) and filter it, you can get a light yellow, translucent AMO/NSP flake silicate mixture, the organic/inorganic ratio (organic/inorganic, O/I) of this mixture is about 40/60.

(e). Add the above AMO/NSP mixture to ethanol (10L) and stir evenly and equivalent NaOH, then add water (10L) and stir evenly, and finally add toluene (10L) and mix well. After standing for one day, it is divided into three layers, the top layer is toluene and AMO (recyclable), the middle layer is ethanol, and the bottom layer is the desired product NSP suspension.

2. Anionic surfactant: Sodium dodecyl sulfonate (SDS) is used in the examples.

i. Simulated virus: A cationic surfactant is used to simulate a virus with a positive charge on the surface. The example uses tallow alkyl amine + HCl . Other cationic surfactants that can be used include dimethyl benzyl ammonium chloride, alkyl(C32)trimethyl ammonium chloride, hexadecyl Trimethyl ammonium chloride (cetyl trimethyl ammonium chloride), dodecyl dimethyl ammonium chloride (dodecyl dimethyl ammonium chloride), SINOTEX ^® series (Central Japan Synthetic Chemical Company, quaternary ammonium salt, poly(quaternary distearyl amidoammonium) chloride)).

The operation steps of the preferred embodiment of the present invention are as follows.

Example 1:

Test tube (A): Simulate NSP anti-virus test

i. Prepare NSP aqueous solution (0.05wt%, 8mL).

ii. Water bath 50℃~80℃, add simulated virus (20wt%, 0.1mL), let stand for 10 minutes, oscillate with ultrasound for 0.5~1.0 hours, and then stand for 10 minutes.

Test tube (B): Simulate NSP/SDS anti-virus test

i. Prepare NSP aqueous solution (0.05wt%, 8mL), add SDS (20wt%, 0.1mL), The weight ratio of NSP/SDS is 15/85. After standing for 10 minutes, shake with ultrasound for 0.5 to 1.0 hours, and then stand for 10 minutes.

ii. Water bath 50℃~80℃, add simulated virus (20wt%, 0.1mL), let stand for 10 minutes, shake with ultrasound for 0.5~1.0 hours, and then stand for 10 minutes.

Example 2

Test tube (A): pure NSP solution

i. Prepare NSP aqueous solution (0.05wt%, 10mL).

Test tube (B): Simulate NSP anti-virus test

i. Prepare NSP aqueous solution (0.05wt%, 10mL).

ii. Add simulated virus (20wt%, 60μL), water bath 50℃~80℃, ultrasonic vibration for 0.5~1.0 hours.

Test tube (C): Simulate NSP/SDS anti-virus test

i. Prepare NSP aqueous solution (0.05wt%, 500ppm, 10mL), add SDS (20wt%, 60μL), the weight ratio of NSP/SDS is about 29/71.

ii. Add simulated virus (20wt%, 60μL), water bath 50℃~80℃, ultrasonic vibration for 0.5~1.0 hours.

Figure 2 shows the result of Example 1, where the solution in the test tube (A) is uniform and translucent, while the upper half of the test tube (B) is white and the lower half is relatively transparent. The white color indicates that the nano-silicon wafer interacts with cations to condense/fix the movement of the virus. Therefore, it can be expected that the nano-silicon wafer can effectively capture the virus and make it inactive.

Figure 3 shows the results of Example 2, where the pure NSP solution in the test tube (A) is in a uniform and transparent state, and the test tube (B) is in a uniform and translucent state, similar to the test tube (A) in Example 1. The test tube (C) is uniformly white. The appearance of white indicates that the nano-silicon wafer interacts with cations, so it can be expected that the nano-silicon wafer can effectively capture the virus.

In order to increase the dispersibility of NSP in water, non-ionic surfactants can also be added as appropriate, such as Triton ^® X-100 (TEDIA, octylphenol polyethoxylate), SINOPOL ^® series (synthesized in China and Japan) Chemical company, polyethylene glycol ether, polyoxyethylene ether), etc.

Since nano-silicon wafers can be uniformly dispersed in water, different concentrations can be configured depending on the situation. For example: 3wt%~10wt% is a gel form that is convenient for transportation, and when diluted to 0.01wt%~0.1wt%, it can be used as a dry hand or disinfectant spray.

Claims (12)

A method for resisting viral infections with nano-silicon wafers, including the following steps: (i) Provide an aqueous solution of nano-silicon wafers, the concentration of the nano-silicon wafers is 0.01wt%~10.0wt%, the nano-silicon wafers are completely delaminated silicic acid clay, and the wafer diameter ratio is (50~300)× (50~300)×1nm ³ , the surface area is 300m ² /g~800m ² /g, the surface structure includes ≡SiOH and ≡SiO ^- Na ⁺ , the cation exchange capacity is 1.0mequiv/g~1.5mequiv/g; and (ii) Bring the nanosilicon wafer into contact with the charged virus, and absorb the virus by one or more of van der Waals force, hydrogen bond, and neutralization of electricity, so as to interrupt the free movement of the virus. Blocking the route of infection may even cause the virus to lose its activity and achieve the purpose of epidemic prevention. The method according to claim 1, wherein the virus is positively charged. The method according to claim 1, wherein the concentration of the nano-silicon wafer is 0.01 wt% to 0.1 wt%. The method according to claim 1, wherein the aqueous nanosilicon wafer solution further includes an anionic surfactant. The method according to claim 4, wherein the anionic surfactant is sodium dodecyl/lauryl sulfonate (SDS/SLS), sodium stearate (sodium octadecanoate), dextran Sulfuric acid (dextran sulfate sodium, DSS), C12-14 alkyl ethoxy sulfate ((C12-14)alkyl ethoxy sulfate), C10-16 alkylbenzenesulfonic acid ((C10-C16)alkylbenzenesulfonic acid), Sodium dodecylbenzenesulphonate (SDBS), sodium xylene sulfonate (sodium xylene sulfonate), sodium toluene sulfonate (sodium toluene sulfonate), sodium cumene sulfonate (sodium cumene sulfonate), alkyl Calcium alkyl benzene sulfonate or Sodium bis-naphthalene sulfonate. The method according to claim 4, wherein the anionic surfactant is sodium dodecyl sulfonate (SDS). The method according to claim 4, wherein the weight ratio of the NSP to the anionic surfactant is 10/90 to 90/10. The method according to claim 4, wherein the weight ratio of the NSP to the anionic surfactant is 10/90~70/30. The method according to claim 1, wherein the aqueous nanosilicon wafer solution further includes a non-ionic surfactant. The method according to claim 1, wherein the aqueous solution of nano-silicon wafers is in direct contact with charged bacteria or viruses. The method according to claim 1, wherein the aqueous solution of nano-silicon wafers is contacted with charged bacteria or viruses in a spray form. The method according to claim 1, wherein the aqueous solution of nano-silicon wafers is first attached to the surface of an object, and then comes into contact with charged bacteria or viruses.

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