Antibacterial gels and methods of making and using the same

Abstract

The present application provides an antibacterial gel, a preparation method and application thereof. Specifically, the antibacterial gel comprises water and hydrogen or a hydrogen donor. The gel uses hydrogen as an antibacterial and anti-inflammatory effective component. The process preparation method is simple, convenient to use, has strong biological adhesion, long retention time, can sustainably and stably release and lock hydrogen, and has antibacterial effect on various pathogenic bacteria.

Description

Technical Field

[0001] The present invention relates to the field of medicine, in particular to an antibacterial gel and a preparation method and application thereof. Background Art

[0002] The discovery of penicillin by British scientist Fleming in 1928 marked the beginning of humanity's long-standing battle with bacteria. Antibiotics are natural products or their semi-synthetic derivatives produced by microorganisms. These chemicals persist in the natural environment for a long time, forcing bacteria to evolve resistance to them to survive.

[0003] Antibiotic resistance refers to the ability of bacteria and other microorganisms to block the effects of antibiotics and other drugs, rendering standard treatments ineffective. The development of antibiotic resistance is primarily linked to the overuse and misuse of antibiotics. Antibiotic resistance causes hundreds of thousands of deaths annually, and as bacterial resistance develops, the death toll from antibiotic resistance is expected to continue to rise. Antibiotic resistance has become a major problem, posing a serious threat to human life and economic well-being.

[0004] Drug resistance of pathogens has become an important factor affecting the effectiveness of clinical anti-infection. These highly resistant strains have put clinical anti-infection treatment in a difficult situation, and even "no drugs are available". Multidrug-resistant bacteria are difficult to treat, have high mortality rates and are extremely prone to outbreaks, making multidrug-resistant bacteria a focus of hospital infection prevention and control. According to domestic and foreign reports, the problem of infection caused by multidrug-resistant bacteria is becoming increasingly serious. Traditional antibiotics mainly target enzymes necessary for bacterial growth, such as inhibiting cell wall synthesis, cell membrane function, nucleic acid synthesis, protein synthesis, and metabolic functions. In order to survive, bacteria can easily develop drug resistance. Therefore, the search for new antibacterial materials is imminent. Summary of the Invention

[0005] The present invention provides an antibacterial gel comprising micro-nano magnesium hydride and poloxamer gel, characterized by using hydrogen released by the micro-nano magnesium hydride (MgH2) as an effective antibacterial and anti-inflammatory ingredient. When the micro-nano magnesium hydride encounters liquid, it immediately absorbs moisture and undergoes a reaction: MgH2 + 2H2O → Mg(OH)2 + 2H2↑. The preparation process is simple, easy to use, exhibits strong bioadhesion, a long retention time, and can continuously and stably release and lock hydrogen. The gel exhibits temperature-dependent properties, being liquid at low temperatures and gradually gelling as the temperature rises. It has a high hydrogen loading capacity, allows for controlled, long-term hydrogen release, and exhibits high biosafety. It also exhibits antibacterial effects against a variety of pathogens.

[0006] In a first aspect of the present invention, an antibacterial gel is provided, comprising a gel carrier and an antibacterial component, wherein the antibacterial component comprises hydrogen dispersed in the gel carrier, and the hydrogen is generated by the reaction of a hydrogen donor (such as a hydride) and water dispersed in the gel carrier.

[0007] In another preferred embodiment, the gel carrier is prepared by mixing a gelling agent and water.

[0008] In another preferred embodiment, the hydrogen content in the antibacterial gel is ≥100 ppb, preferably ≥200 ppb, more preferably ≥400 ppb, for example ≥500 ppb, ≥600 ppb.

[0009] In another preferred embodiment, the hydrogen content in the antibacterial gel is ≤1000 ppb, preferably ≤800 ppb.

[0010] In another preferred embodiment, the hydrogen donor in the antibacterial gel begins to react with water to generate hydrogen, and continues to react to generate hydrogen within the effective time. The effective time is calculated from the time the hydrogen donor contacts water.

[0011] In another preferred embodiment, the antibacterial component includes hydrogen bubbles dispersed in the gel carrier.

[0012] In another preferred embodiment, the hydrogen in the gel carrier includes hydrogen bubbles in the form of bubbles (micro bubbles) and / or hydrogen in the gel carrier in a dissolved form.

[0013] In another preferred embodiment, the hydride in the antibacterial gel reacts with water to generate hydrogen and the hydrogen content in the antibacterial gel is maintained above an effective concentration (the concentration required to inhibit the growth or reproduction of microorganisms) within an effective time (the time required to inhibit the growth or reproduction of microorganisms).

[0014] In another preferred embodiment, the effective time is ≥1h, preferably ≥2h, preferably ≥5h, preferably ≥10h, more preferably ≥20h, such as ≥30h, ≥40h, ≥50h, ≥60h.

[0015] In another preferred embodiment, the effective time is ≤100h.

[0016] In another preferred embodiment, the effective concentration of hydrogen in the antibacterial gel is ≥100 ppb, preferably ≥200 ppb, more preferably ≥400 ppb, for example ≥500 ppb, ≥600 ppb.

[0017] In another preferred embodiment, the effective concentration of hydrogen in the antibacterial gel is ≤1000 ppb, preferably ≤800 ppb.

[0018] In another preferred embodiment, the hydride is selected from lithium hydride (LiH), palladium hydride (PdH), beryllium hydride (BeH2), magnesium hydride (MgH2), calcium hydride (CaH2), strontium hydride (SrH2), titanium hydride (TiH2), aluminum hydride (AlH3), boron hydride (BH3), lithium borohydride (LiBH4), sodium borohydride (NaBH4), potassium borohydride (KBH4), magnesium borohydride (Mg(BH4)2), calcium borohydride (Ca(BH4)2), lithium aluminum hydride (LiAlH4), sodium aluminum hydride (NaAlH4), potassium aluminum hydride (KAlH4), magnesium aluminum hydride (Mg(AlH4)2), calcium aluminum hydride (Ca(AlH4)2), ammonia borane (NH3BH3), lithium amide (LiNH2), lithium imide (Li2NH) and mixtures thereof.

[0019] In another preferred embodiment, the hydrogen donor is lithium hydride, calcium hydride, palladium hydride, or magnesium hydride; preferably magnesium hydride.

[0020] In another preferred embodiment, the hydride is a micro-nano hydride, preferably micro-nano magnesium hydride.

[0021] In another preferred embodiment, the particle size of the micro-nano hydride (such as micro-nano magnesium hydride) is 0.1-100 μm.

[0022] In another preferred embodiment, the gelling agent is hydroxyethyl cellulose, hydroxymethyl cellulose, hydroxypropyl guar gum, methyl cellulose, ethyl cellulose, hydroxypropyl cellulose, sodium carboxymethyl cellulose, carbomer, alginate, gelatin, agar, chitosan, poly N-isopropylacrylamide and poloxamer; preferably poloxamer.

[0023] In another preferred embodiment, the antibacterial gel comprises poloxamer gel and hydride, wherein the poloxamer gel and hydride are mixed together or exist separately.

[0024] In another preferred embodiment, the poloxamer is selected from the group consisting of poloxamer 407, poloxamer 188, poloxamer 184, or a combination thereof.

[0025] In another preferred embodiment, the weight percentage of the poloxamer is 15%-28% (w / v), preferably 20%-25% (w / v), and more preferably 23%-25% (w / v), based on the total volume of the antibacterial gel.

[0026] In another preferred embodiment, the concentration of the hydride is 0.2 mg / mL-5 mg / mL, preferably 0.5 mg / mL-3 mg / mL, based on the total volume of the antibacterial gel.

[0027] In another preferred embodiment, the antibacterial gel comprises or consists of the following components: 15%-28% (w / v) poloxamer, 0.2 mg / mL-3 mg / mL micro-nano magnesium hydride, and the remainder water or physiological saline, based on the total volume of the antibacterial gel.

[0028] In another preferred embodiment, the antibacterial component further contains antibiotics. Preferably, the antibiotics are selected from: glycopeptide antibiotics, β-lactam antibiotics, aminoglycoside antibiotics, quinolone antibiotics, polypeptide antibiotics or a combination thereof.

[0029] In another preferred embodiment, the concentration of the antibiotic (such as vancomycin, polymyxin B) is 0.1-5 μg / mL (preferably 0.2-2 μg / mL), based on the total volume of the antibacterial gel.

[0030] The second aspect of the present invention provides a method for preparing an antibacterial gel, comprising the steps of:

[0031] (1) providing a gel (preferably a poloxamer gel, more preferably a poloxamer content of 15% to 28% (w / v) in the gel);

[0032] (2) Mixing hydrogen or a hydrogen donor into the gel (the hydrogen donor is a hydride, and preferably the concentration of the hydride is 0.2 mg / mL-3 mg / mL) to obtain an antibacterial gel.

[0033] In another preferred embodiment, the hydrogen donor is micro-nano magnesium hydride.

[0034] In another preferred embodiment, poloxamer is dissolved in water or 0.9% sodium chloride solution to obtain poloxamer gel. Preferably, the dissolution is carried out at a low temperature (eg, 0-4°C).

[0035] In another preferred embodiment, step (1) further comprises sterilizing the gel.

[0036] In another preferred embodiment, step (2) is performed by vortex mixing or stirring mixing at low temperature (eg, 0-4°C).

[0037] In another preferred embodiment, the method further comprises the step of: (3) adding antibiotics to the antibacterial gel.

[0038] The third aspect of the present invention provides a preparation, which contains the antibacterial gel according to the first aspect of the present invention, or the preparation includes:

[0039] (1) a first container and a gel (preferably a poloxamer gel) placed in the first container; and

[0040] (2) A second container and hydrogen or a hydrogen donor (preferably, the hydrogen donor is a hydride) placed in the second container. When in use, the hydrogen or hydrogen donor is mixed evenly with the gel.

[0041] In another preferred embodiment, the preparation further comprises a third container and an antibiotic placed in the third container.

[0042] In another preferred embodiment, any two or three of the first container, the second container and the third container can be combined into one.

[0043] In another preferred embodiment, the poloxamer gel is a gel obtained by dissolving poloxamer in water or 0.9% sodium chloride solution. Preferably, the weight percentage of the poloxamer is 15%-28% (w / v), preferably 20%-25% (w / v), and more preferably 23%-25% (w / v), based on the total volume of the poloxamer gel.

[0044] In another preferred embodiment, the poloxamer is selected from the group consisting of poloxamer 407, poloxamer 188, poloxamer 184, or a combination thereof.

[0045] In another preferred embodiment, the hydride is micro-nano magnesium hydride. Preferably, the particle size of the micro-nano magnesium hydride is 10 nm-100 μm.

[0046] In another preferred embodiment, the concentration of magnesium hydride is 0.2 mg / mL-3 mg / mL, preferably 0.5 mg / mL-2 mg / mL, based on the total volume of the poloxamer gel.

[0047] The fourth aspect of the present invention provides the use of the antibacterial gel described in the first aspect of the present invention for preparing antibacterial drugs (such as antibacterial gel drugs).

[0048] In another preferred embodiment, the bacteria include Gram-positive bacteria and / or Gram-negative bacteria.

[0049] In a fifth aspect, the present invention provides a composition comprising a first active ingredient and a second active ingredient, wherein the first active ingredient is hydrogen or a hydrogen donor, and the second active ingredient is an antibiotic.

[0050] In another preferred embodiment, the first active component is a hydrogen donor, and the hydrogen donor is a hydride.

[0051] In another preferred embodiment, the hydride is selected from lithium hydride (LiH), palladium hydride (PdH), beryllium hydride (BeH2), magnesium hydride (MgH2), calcium hydride (CaH2), strontium hydride (SrH2), titanium hydride (TiH2), aluminum hydride (AlH3), boron hydride (BH3), lithium borohydride (LiBH4), sodium borohydride (NaBH4), potassium borohydride (KBH4), magnesium borohydride (Mg(BH4)2), calcium borohydride (Ca(BH4)2), lithium aluminum hydride (LiAlH4), sodium aluminum hydride (NaAlH4), potassium aluminum hydride (KAlH4), magnesium aluminum hydride (Mg(AlH4)2), calcium aluminum hydride (Ca(AlH4)2), ammonia borane (NH3BH3), lithium amide (LiNH2), lithium imide (Li2NH) and mixtures thereof.

[0052] In another preferred embodiment, the hydrogen donor is lithium hydride, calcium hydride, palladium hydride, or magnesium hydride; preferably magnesium hydride.

[0053] In another preferred embodiment, the first active ingredient and / or the second active ingredient is dissolved and / or suspended in a gel, and the gel is hydroxyethyl cellulose, hydroxymethyl cellulose, hydroxypropyl guar gum, methyl cellulose, ethyl cellulose, hydroxypropyl cellulose, sodium carboxymethyl cellulose, carbomer, alginate, gelatin, agar, chitosan, poly N-isopropylacrylamide and poloxamer; preferably poloxamer gel.

[0054] The sixth aspect of the present invention provides a medicine comprising the composition according to the fifth aspect of the present invention and a pharmaceutically acceptable carrier.

[0055] In another preferred embodiment, the drug is a gel preparation, in which hydrogen is dissolved and / or suspended and dispersed in the gel.

[0056] In another preferred embodiment, the antibiotic is dissolved in the gel.

[0057] The seventh aspect of the present invention provides the use of the composition described in the fifth aspect of the present invention for preparing a medicament for killing microorganisms.

[0058] An eighth aspect of the present invention provides a medicine kit, comprising:

[0059] (i) a container a, and hydrogen gas, or a hydrogen gas donor, placed in the container a; and

[0060] (ii) a container b, and an antibiotic placed in the container b.

[0061] In another preferred embodiment, the container a and the container b can be combined into one.

[0062] In another preferred embodiment, the antibiotic in the container b is a gel containing antibiotics.

[0063] According to a ninth aspect of the present invention, a method for killing microorganisms is provided, comprising the steps of applying hydrogen to living microorganisms.

[0064] In another preferred embodiment, the method comprises the step of applying hydrogen gas and antibiotics to the living microorganisms.

[0065] In another preferred embodiment, the method is a non-therapeutic method, for example, used to kill microorganisms in the environment.

[0066] In another preferred embodiment, the method is a therapeutic method, for example, for disinfecting wound surfaces.

[0067] In another preferred embodiment, the method comprises the step of applying an antibacterial gel containing hydrogen to the living microorganisms.

[0068] In another preferred embodiment, the hydrogen content in the antibacterial gel is ≥100 ppb, preferably ≥200 ppb, more preferably ≥400 ppb, for example ≥500 ppb, ≥600 ppb.

[0069] In another preferred embodiment, the hydrogen content in the antibacterial gel is ≤1000 ppb, preferably ≤800 ppb.

[0070] In another preferred embodiment, hydrogen is generated by the reaction of hydride and water in the antibacterial gel and the hydrogen content in the antibacterial gel is maintained above an effective concentration (the concentration required to inhibit the growth or reproduction of microorganisms) within an effective time (the time required to inhibit the growth or reproduction of microorganisms).

[0071] In another preferred embodiment, the effective time is ≥1h, preferably ≥2h, preferably ≥5h, preferably ≥10h, more preferably ≥20h, such as ≥30h, ≥40h, ≥50h, ≥60h.

[0072] In another preferred embodiment, the effective time is ≤100h.

[0073] In another preferred embodiment, the effective concentration of hydrogen in the antibacterial gel is ≥100 ppb, preferably ≥200 ppb, more preferably ≥400 ppb, for example ≥500 ppb, ≥600 ppb.

[0074] In another preferred embodiment, the effective concentration of hydrogen in the antibacterial gel is ≤1000 ppb, preferably ≤800 ppb.

[0075] In a tenth aspect, the present invention provides a method for enhancing the activity of antibiotics in killing microorganisms, the method comprising the steps of simultaneously using antibiotics and hydrogen to act on living microorganisms, thereby killing the microorganisms.

[0076] In another preferred embodiment, the microorganism is an antibiotic-resistant microorganism.

[0077] In another preferred embodiment, the method comprises dissolving and / or suspending hydrogen or a hydrogen donor in an antibiotic-containing gel and then allowing the gel to act on living microorganisms, wherein the hydrogen donor is a hydride, and preferably the gel is a poloxamer gel.

[0078] The present invention also provides a method for improving the antibacterial activity of magnesium hydride, comprising mixing micro-nano magnesium hydride with poloxamer gel.

[0079] The present invention also provides the use of hydrogen for:

[0080] (1) Inhibit or kill microorganisms;

[0081] (2) Preparation of drugs that inhibit or kill microorganisms.

[0082] In another preferred embodiment, the drug is a gel preparation.

[0083] In another preferred embodiment, the medicine further comprises antibiotics.

[0084] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features described in detail below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be listed here one by one.

[0085] The following will further explain the concept, specific structure and technical effects of the present invention in combination to fully understand the purpose, characteristics and effects of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0086] Figure 1 It shows that the gel has temperature-dependent properties;

[0087] Figure 2 The slow release of hydrogen from the nanomagnesium hydride gel was shown;

[0088] Figure 3 Shown are the bacterial cells observed under a transmission electron microscope. DETAILED DESCRIPTION

[0089] Through extensive and in-depth research, the inventors unexpectedly discovered that directly mixing micro-nano magnesium hydride powder with poloxamer gel can achieve a "hydrogen lock" effect, enabling magnesium hydride, which has no antibacterial effect against aerobic bacteria in a gel-free state, to exert its antibacterial effect in the gel. The micro-nano magnesium hydride gel of the present invention is simple to prepare, continuously releases and locks hydrogen, is easy to use, has strong bioadhesion, and has a long retention time. Through the gel preparation process, micro-nano magnesium hydride, which originally had no antibacterial activity, is transformed into a material with antibacterial activity.

[0090] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0091] As used herein, when used in reference to a specific recited value, the term "about" means that the value may vary by no more than 1% from the recited value. For example, as used herein, the expression "about 100" includes all values ​​between 99 and 101 (e.g., 99.1, 99.2, 99.3, 99.4, etc.).

[0092] As used herein, the terms "comprising" or "including" may be open, semi-closed, or closed. In other words, the terms also include "consisting essentially of" or "consisting of."

[0093] Hydrogen and hydrogen donors

[0094] Hydrogen (H2), the smallest molecule in nature, possesses strong reducing properties and high permeability. It is a newly discovered biogas signaling molecule and is considered an excellent antioxidant molecule that is non-toxic to normal cells. It has been reported to have broad application prospects in the treatment of many diseases, including Alzheimer's disease, stroke, and cancer. However, there are no reports of direct use of hydrogen for antibacterial purposes. Traditional methods of hydrogen administration often have drawbacks, such as difficult storage, easy release, and lack of targeting capabilities.

[0095] The term "hydrogen donor" in the present invention is defined as a substance that can provide hydrogen when needed, especially a substance that can react with water to generate hydrogen. Typical hydrogen donors include hydrides. In a preferred embodiment of the present invention, the hydride is selected from lithium hydride (LiH), palladium hydride (PdH), sodium hydride (NaH), potassium hydride (KH), beryllium hydride (BeH2), magnesium hydride (MgH2), calcium hydride (CaH2), strontium hydride (SrH2), titanium hydride (TiH2), aluminum hydride (AlH3), boron hydride (BH3), lithium borohydride (LiBH4), sodium borohydride (NaBH4), potassium borohydride (KBH4), magnesium borohydride (Mg(BH4)2), calcium borohydride (Ca(BH4)2), lithium aluminum hydride (LiAlH4), sodium aluminum hydride (NaAlH4), potassium aluminum hydride (KAlH4), magnesium aluminum hydride (Mg(AlH4)2), calcium aluminum hydride (Ca(AlH4)2), ammonia borane (NH3BH3), lithium amide (LiNH2), lithium imide (Li2NH) and a mixture of two or more thereof.

[0096] In a preferred embodiment of the present invention, the hydride is a micro-nano hydride. The term "micro-nano hydride" as used herein is defined as hydride particles having a particle size of 1 nm to 100 μm. Preferably, the particle size of the micro-nano hydride is 5 nm to 50 μm; more preferably, the particle size of the micro-nano hydride is 10 nm to 10 μm. Micro-nano hydrides can be purchased commercially or prepared in the laboratory using known methods.

[0097] antibiotic

[0098] As used herein, the term "antibiotic" is defined as a substance that inhibits the growth of microorganisms without harming the host. For example, antibiotics can inhibit cell wall synthesis, protein synthesis, nucleic acid synthesis, or alter cell membrane function. As used herein, "antibiotics" include antibiotics extracted from microbial culture fluids as well as chemically synthesized or semi-synthesized antibiotics.

[0099] Classes of antibiotics include, but are not limited to, polymyxins (polymyxin B, polymyxin E), macrolides (e.g., erythromycin), penicillins (e.g., nafcillin), cephalosporins (e.g., cefazolin), carbapenems (e.g., imipenem), monobactams (e.g., aztreonam), other β-lactam antibiotics, oxalines (e.g., linezolid), aminoglycosides (e.g., gentamicin, ethidium), chloramphenicol, and cyclophosphamides (e.g., sulfamethoxazole). , glycopeptides (e.g., vancomycin), quinolones (e.g., ciprofloxacin), cyclic lipopeptide antibiotics (e.g., daptomycin), tetracyclines (e.g., minocycline), fusidic acid, trimethoprim, metronidazole, clindamycin, mupirocin, rifamycins (e.g., rifampicin), streptococcins (e.g., quinupristin and dalfopristin), lipoproteins (e.g., daptomycin), polyenes (e.g., amphotericin B), azoles (e.g., fluconazole), and echinocandins (e.g., caspofungin acetate).

[0100] Examples of specific antibiotics include, but are not limited to, polymyxin B, colistin, erythromycin, nafcillin, cefazolin, imipenem, aztreonam, gentamicin, sulfamethoxazole, vancomycin, ciprofloxacin, trimethoprim, rifampicin, metronidazole, clindamycin, teicoplanin, mupirocin, azithromycin, clarithromycin, daptomycin, ofloxacin, lomefloxacin, norfloxacin, nalidixic acid, sparfloxacin, pefloxacin, amlofloxacin, gatifloxacin, moxifloxacin, gemifloxacin, enoxacin, fleroxacin, minocycline, linezolid, temafloxacin, tofaxan, clinafloxacin, sulbactam, clavulanic acid, amphotericin B, fluconazole, itraconazole, ketoconazole, and nystatin.

[0101] In a preferred embodiment, the antibiotics include: vancomycin, polymyxin B, linezolid, and daptomycin.

[0102] Composition

[0103] Herein, the composition comprises a first active ingredient and a second active ingredient, wherein the first active ingredient is hydrogen or a hydrogen donor, and the second active ingredient is an antibiotic.

[0104] In a preferred embodiment of the present invention, the first active ingredient is a hydrogen donor, and the hydrogen donor is a hydride, preferably magnesium hydride.

[0105] In the composition of the present invention, the first active ingredient and / or the second active ingredient are dissolved and / or suspended in a gel, preferably a poloxamer gel. The combination of hydrogen and antibiotics in the composition of the present invention can enhance antibacterial efficacy, reduce the dosage of antibiotics while achieving the same or even better antibacterial efficacy, and reduce the risk of bacterial resistance.

[0106] gel

[0107] As used herein, a gel refers to a colloidal system composed of a liquid and a solid in a thickened liquid, semi-solid, or solid state. A gel can be a composition that is physically cross-linked through entangled polymer chains or an interconnected network, or a composition that is chemically cross-linked through covalent bonds, such that it swells but does not dissolve in a liquid.

[0108] Gels are typically obtained using a gelling agent (gelling matrix). As used herein, the term "gelling agent" refers to a polymer that can form a gel when dispersed in any suitable liquid or semisolid substance. As used herein, the term "polymer" includes homopolymers and copolymers. A homopolymer is a polymer formed by the polymerization of one monomer, while a copolymer is a polymer formed by the polymerization of two or more monomers. A "block copolymer" refers to a copolymer in which similar monomer units appear in relatively long, alternating sequences along the chain. As used herein, the term "gel composition" refers to a gelling agent dispersed, dissolved, or swollen in a suitable liquid (preferably water) or semisolid substance.

[0109] In certain embodiments, the gelling agent is a pharmaceutically acceptable gelling agent.In preferred embodiments, the gelling agent forms a gel when dissolved or suspended in an aqueous liquid.

[0110] In certain embodiments, the gelling agent is present in the gel at a concentration of between about 0.1% and 10% w / v, and in some embodiments, the gelling agent is present in the gel at a concentration of between about 0.5% and 5% w / v. In certain preferred embodiments, the gelling agent is present in the gel at a concentration of between about 1% and 3% w / v, and in a preferred embodiment thereof, the gelling agent is present in the gel at a concentration of between about 1% and 2% w / v.

[0111] In a preferred embodiment, the gelling agent is selected from the group consisting of hydroxyethyl cellulose, hydroxymethyl cellulose, hydroxypropyl guar gum, methyl cellulose, ethyl cellulose, hydroxypropyl cellulose, sodium carboxymethyl cellulose, carbomer, alginate, gelatin, agar, chitosan, poly (N-isopropylacrylamide) and poloxamer.

[0112] drug

[0113] Herein, the medicine includes the composition of the present invention and a pharmaceutically acceptable carrier.

[0114] In a preferred embodiment, the drug is a gel preparation, in which hydrogen is dissolved and / or suspended and dispersed in the gel. In a preferred embodiment, the antibiotic is dissolved in the gel.

[0115] medicine box

[0116] The term "kit" as used herein refers to any delivery system for delivering materials that allows for the storage, transport or delivery of appropriate reagents (e.g., hydrogen donors, gels, antibiotics, sterile water, etc. in appropriate containers) and / or devices (e.g., swabs) and / or supporting materials (e.g., written instructions for use, etc.).

[0117] For example, a kit includes one or more accessories (e.g., boxes, bags) containing relevant reagents and / or support materials. As used herein, the term "segmented kit" refers to a delivery system comprising two or more different containers, each containing a sub-portion of the total kit components. The containers can be delivered together or individually to a desired recipient. For example, a first container can contain a hydrogen donor for a specific use, a second container can contain a gel for a specific use (which can have antibiotics pre-dissolved in the gel), and a third container can contain a sterile liquid such as water or a buffer.

[0118] In fact, any delivery system containing two or more different containers, each containing a sub-portion of the total kit components, is included in the term "segmented kit". In contrast, a "combination kit" refers to a delivery system that contains all the components of the reaction materials required for a particular use in a single container (e.g., in a single box containing each desired component). The term "kit" includes both segmented and combination kits.

[0119] application

[0120] The medicament of the present invention can be used to kill microorganisms.

[0121] As used herein, the term "microorganism" refers to any species or type of microorganism, including but not limited to bacteria, archaea, fungi, protozoa, mycoplasmas, and parasites. The present invention contemplates that many of the microorganisms contained therein are also pathogenic to the subject.

[0122] The term "bacteria" refers to all prokaryotes, including those in all phyla in the prokaryotic kingdom. The term is intended to include all microorganisms that are considered to be bacteria, including mycoplasma, Chlamydia, Actinomyces, Streptomyces, and Rickettsia. All forms of bacteria are included in this definition, including cocci, bacilli, spirochetes, spheroplasts, protoplasts, etc. Also included in this term are gram-negative or gram-positive prokaryotes. "Gram-negative" and "Gram-positive" refer to the staining patterns using Gram staining methods, which are well known in the art (see, for example, Finegold and Martin, Diagnostic Microbiology, 6th edition, CV Mosby St. Louis, 13-15 pages (1982)). "Gram-positive bacteria" are bacteria that retain the primary dye used in the Gram stain, resulting in the stained cells typically appearing dark blue to purple under the microscope. "Gram-negative bacteria" are bacteria that do not retain the primary dye used in the Gram stain but are stained by a counterstain. Therefore, Gram-negative bacteria typically appear red.

[0123] As used herein, the term "fungus" is used to refer to eukaryotic organisms such as molds and yeasts, including dimorphic fungi.

[0124] In another preferred embodiment, the "microorganism" is a drug-resistant pathogen.

[0125] In some embodiments, the antibacterial gel of the present invention further comprises one or more antibiotics. In one embodiment, the antibiotics are selected from vancomycin, polymyxin B, or a combination thereof. The antibacterial gel of the present invention, when used in combination with antibiotics, can enhance antibacterial efficacy, reduce the dosage of antibiotics while achieving the same or even better antibacterial effect, and reduce the risk of bacterial resistance.

[0126] The present invention also provides a method for increasing the antibacterial activity of magnesium hydride. By mixing micro-nano magnesium hydride with poloxamer gel, the poloxamer gel effectively locks in the hydrogen released by the magnesium hydride, preventing bacteria from generating normal energy, thereby directly achieving a bactericidal effect. Furthermore, the magnesium hydride used in the antibacterial gel of the present invention is at a relatively low concentration (0.02-0.2% w / v, i.e., 0.2-2 mg / mL). Therefore, there is no need to alter the pH of the environment to achieve sterilization. The locked hydrogen directly inhibits bacterial ATP production, achieving a strong antibacterial effect.

[0127] If necessary, the preparation of the present invention may further contain appropriate solubilizers, isotonic agents, diluents, excipients, pH adjusters, analgesics, antioxidants, etc. Solubilizers include, for example, polyoxyethylene hydrogenated castor oil, polysorbate 80, nicotinamide, polyoxyethylene sorbitan monolaurate, polyethylene glycol, and castor oil fatty acid ethyl ester. Isotonic agents include, for example, sodium chloride, potassium chloride, and calcium chloride.

[0128] Compared with existing methods, it has the following advantages:

[0129] 1. Micro-nano magnesium hydride gel is simple to prepare, continuously releases and locks hydrogen, is easy to use, has strong bioadhesion, and has a long retention time. Through the gel preparation process, micro-nano magnesium hydride, which originally had no antimicrobial activity, is transformed into a material with antimicrobial activity.

[0130] 2. The combination of the micro-nano magnesium hydride gel of the present invention and antibiotics can enhance the antibacterial effect, reduce the dosage of antibiotics, and thus reduce the risk of bacterial resistance.

[0131] 3. The concentration of micro-nano magnesium hydride used in this method is relatively low, and there is no need to sterilize by changing the environmental pH. The locked hydrogen can directly inhibit the production of bacterial ATP, thereby achieving the purpose of sterilization, with higher safety and antibacterial effect.

[0132] 4. The antibacterial gel of the present invention has temperature-dependent properties. It is liquid at low temperatures and gradually becomes gel-like as the temperature rises. It has a high hydrogen loading capacity, can regulate the long-term slow release of hydrogen, has high biosafety, and has antibacterial effects against a variety of pathogens.

[0133] The present invention is further described below with reference to specific implementation cases. The following examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. Therefore, the scope of protection of the patent of the present invention shall be subject to the appended claims. The experimental materials and reagents used in the following examples can be obtained from commercial channels unless otherwise specified. Normal temperature or room temperature refers to 4°C-25°C, preferably 15-25°C.

[0134] Example 1 Preparation of different gel matrices

[0135] Add 20-25% by mass of Poloxamer 407 to 0.9% sodium chloride solution, stir and place at 4°C overnight to dissolve, and sterilize by high pressure steam after complete dissolution. Figure 1 It is liquid at low temperatures and gradually turns into a gel as the temperature rises.

[0136] Different amounts of nanomagnesium hydride were weighed and added to the gel, vortex-mixed at low temperature, to obtain magnesium hydride gels. 1.5% agar was used as a coagulant control. The gels with different matrices were compared and evaluated in terms of viscosity, coagulation time, and skin adhesion. The results are shown in Table 1. It can be seen that the 23% poloxamer group effectively reduced gel viscosity and improved skin adhesion compared to the 1.5% agar control group; the 25% poloxamer group reduced gel viscosity, shortened coagulation time, and improved skin adhesion compared to the 1.5% agar control group.

[0137] Table 1

[0138]

[0139] Example 2 Preparation of Nano-Magnesium Hydride Gel and Analysis of Its Anti-Gram-Positive Bacteria Activity

[0140] This embodiment relates to an antibacterial gel, wherein the micro-nano magnesium hydride is mixed in poloxamer 407.

[0141] Add 23% (w / v) Poloxamer 407 to a 0.9% sodium chloride solution, stir, and dissolve overnight at 4°C. After complete dissolution, sterilize with high-pressure steam. Add nanomagnesium hydride with a particle size of 10-100 nm to the gel and vortex mix at low temperature to prepare magnesium hydride gels of varying concentrations.

[0142] The hydrogen in the nanomagnesium hydride gel prepared by this method can be released slowly. The maximum release amount of about 600 ppb of nanomagnesium hydride at 0.4 mg / mL is reached after 24 hours, and the hydrogen release process can last up to 72 hours. In contrast, if the same amount of nanomagnesium hydride is only in normal saline, the hydrogen is released very quickly, and is basically completely released after 36 hours, with most of the hydrogen gas released into the air. Figure 2 Shows the slow release of hydrogen from nanomagnesium hydride gel.

[0143] The gel was used to determine its activity against Gram-positive bacteria using the gel-mixed culture method. Gram-positive, methicillin-resistant Staphylococcus aureus (MRSA) ATCC 43300 and vancomycin-resistant Enterococcus faecalis (VRE) ATCC 51299 were used as test bacteria for antimicrobial activity. A single colony of MRSA ATCC 43300 was inoculated in 4 mL of LB medium and cultured overnight at 37°C with shaking. VRE ATCC 51299 was inoculated in 4 mL of BHI medium and cultured overnight at 37°C with shaking. The resulting dilution was then inoculated into fresh culture medium at a 1:100 ratio and cultured for another 4 hours, allowing the bacteria to reach the logarithmic growth phase.

[0144] Take a certain amount of bacterial solution and add it to the freshly prepared magnesium hydride gel or physiological saline, mix well, and make the bacterial concentration be 5×10 6CFU / mL. At the same time, the same molar concentration of MgSO4 corresponding to the highest MgH2 concentration in the test was set to exclude Mg 2+ The NaOH group (pH corresponding to the highest MgH2 concentration) was set up to eliminate the effect of pH increase caused by MgH2 dissolving in water on the growth inhibition of bacteria. The cells were placed in a 37°C incubator for 12 to 18 hours, then placed at 4°C to make them liquid. The cells were mixed with a pipette and then diluted in gradients. The bacterial count was determined by the spot plate method.

[0145] The results are shown in Table 2. Under liquid culture conditions without poloxamer gel, micro-nano magnesium hydride had no significant effect on the several aerobic bacteria tested.

[0146] The bacteria were observed under the action of 2 mg / mL MgH2 gel by transmission electron microscopy. Figure 3 The results showed that the cell structure of the control group was intact and clear, with smooth cell walls. The cytoplasm and nuclear regions were clearly visible, and the cell division septum was visible normally. After the magnesium hydride gel treatment, intracellular material was lost, cytoplasm condensed, vacuolation was obvious, and the cells deformed, shrank, and divided abnormally. This shows that magnesium hydride gel has a strong bactericidal effect.

[0147] Through the analysis of differentially expressed genes in the prokaryotic transcriptome, it was found that the effect of nano-magnesium hydride on bacteria was related to the ATP synthesis pathway, causing the related genes of this pathway to be downregulated. After qPCR verification of the differentially expressed genes, it was consistent with the transcriptomics results, indicating that the antibacterial effect of magnesium hydride gel is related to the inhibition of ATP synthesis.

[0148] Table 2

[0149]

[0150] Example 3 Preparation of Micro-Nano Magnesium Hydride Gel and Its Anti-Gram-Negative Bacteria Activity

[0151] This embodiment relates to an antibacterial gel, wherein the micro-nano magnesium hydride is mixed in poloxamer 188.

[0152] Add 20% (w / v) poloxamer 188 to a 0.9% sodium chloride solution, stir, and dissolve overnight at 4°C. After complete dissolution, sterilize with high-pressure steam. Add micro-nano magnesium hydride with a particle size of 500nm-100μm to the cryogel and vortex mix thoroughly. The gel is used to determine anti-Gram-negative bacterial activity using the gel mixed culture method. Pseudomonas aeruginosa ATCC 9027 is used as the anti-Gram-negative bacterial test bacteria, and the specific testing method is the same as in Example 2.

[0153] The results showed that after overnight magnesium hydride gel mixed culture, 1 mg / mL magnesium hydride group could reduce the bacterial concentration by 104 CFU / mL, with a sterilization rate of 99.999%, demonstrating that magnesium hydride gel possesses a strong bactericidal effect. However, the micro-nano magnesium hydride group (not formulated with poloxamer gel) and the blank poloxamer gel control group showed no bactericidal effect. This suggests that poloxamer gel imparts magnesium hydride with strong anti-Gram-negative bacterial activity.

[0154] Example 4 Preparation of Nano-MgH Gel and Anti-MRSA ATCC 43300 on Mouse Embryonic Fibroblast NIH3T3 Cell Surface

[0155] This embodiment relates to an antibacterial gel, wherein the micro-nano magnesium hydride is mixed in poloxamer 184.

[0156] A certain amount of nano-magnesium hydride of 200-1000 nm was weighed and added to a low-temperature sterile 15% poloxamer 184 gel (poloxamer dissolved in DMEM medium) according to the w / v ratio, and vortexed to mix. 200 μL of NIH3T3 cells (2*10 4 cells / well) in DMEM (10% FBS, double antibody), adhere to the wall and culture for 4 hours, remove the supernatant, wash 3 times with PBS; add 1*10 6 100 μL of MRSA bacteria (CFU / well) was added to each well and incubated in a cell culture incubator for 3 hours. The supernatant was removed, and 200 μL of DMEM poloxamer containing varying concentrations of magnesium hydride was added. The cells were then incubated in a cell culture incubator for 18 hours. Cell viability was assessed using CCK-8 reagent, and bacterial counts and sterilization rates were determined using the spot-seeding method.

[0157] CCK-8 cell viability assay results showed that when the MgH2 concentration was ≤2.0 mg / mL, cell viability was virtually unaffected, and the cell surface sterilization rate was positively correlated with the MgH2 concentration. This indicates that magnesium hydride gel has high safety and strong antibacterial activity.

[0158] Table 3

[0159]

[0160] Example 5 Preparation of Vancomycin-Nanomagnesium Hydride Gel and Its Anti-Gram-Positive Bacteria Effect

[0161] This embodiment relates to a nano magnesium hydride antibacterial gel, wherein the magnesium hydride micro-nano powder and vancomycin are mixed in the gel and used together to fight Gram-positive bacteria.

[0162] Nano-magnesium hydride with a particle size of 50-100 nm was added to a low-temperature sterile 20% poloxamer 407 gel at a w / v ratio to make the final concentration of nano-magnesium hydride 1 mg / mL. Stirring was carried out at low temperature to make it uniformly dispersed. Then, a sublethal dose of vancomycin (0.5 μg / mL) was added to prepare vancomycin-nano-magnesium hydride gel. The gel mixed culture method was used, and a MgSO4 group with the same molar concentration corresponding to the 1 mg / mL MgH2 concentration was set to exclude Mg. 2+ The NaOH group (pH corresponding to 1 mg / mL MgH2 concentration) was set to eliminate the effect of the pH increase caused by MgH2 dissolving in water on bacterial growth inhibition, and was combined with 0.5 μg / mL vancomycin. MRSAATCC 43300 was used as the test bacteria for anti-Gram-positive activity, and the specific testing method was the same as in Example 2. The results showed that the antibacterial effect of vancomycin and magnesium hydride was significantly improved compared to vancomycin or magnesium hydride alone, demonstrating that the combination of magnesium hydride and antibiotics can reduce the amount of antibiotic used and the risk of inducing bacterial resistance, while achieving the same antibacterial effect (Table 4).

[0163] Table 4

[0164]

[0165] Example 6 Preparation of polymyxin-nanomagnesium hydride gel and its anti-Gram-negative bacteria effect

[0166] This embodiment relates to a nano-magnesium hydride antibacterial gel, in which the magnesium hydride micro-nano powder is mixed with polymyxin B in the gel (the preparation method is the same as that of Example 5, wherein vancomycin (0.5 μg / mL) is replaced with polymyxin B (0.5 μg / mL)), and is used in combination to fight Gram-negative bacteria. Pseudomonas aeruginosa P.aeruginosa ATCC 9027 was used as the test bacteria for anti-Gram-negative bacteria activity, and the specific test method was the same as that of Example 2. The results showed that the antibacterial effect of polymyxin B and magnesium hydride was significantly improved compared to the use of polymyxin B or magnesium hydride alone, proving that the combination of magnesium hydride and antibiotics can reduce the amount of antibiotics used and reduce the risk of inducing bacterial resistance while achieving the same antibacterial effect (Table 5).

[0167] Table 5

[0168]

[0169]

[0170] The above describes in detail the preferred embodiments of the present invention. It should be understood that numerous modifications and variations based on the concepts of the present invention can be made by those skilled in the art without inventive effort. Therefore, any technical solution that can be derived by a person skilled in the art based on the concepts of the present invention through logical analysis, reasoning, or limited experimentation based on the existing technology should be within the scope of protection defined by the claims.

Claims

1. An antibacterial gel, characterized in that: The antibacterial gel comprises a gel carrier and an antibacterial component, wherein the antibacterial component comprises hydrogen dispersed in the gel carrier, and the hydrogen is generated by the reaction of a hydrogen donor dispersed in the gel carrier and water; The antibacterial component further contains an antibiotic, which is vancomycin or polymyxin B. The hydrogen content in the antibacterial gel is ≥100 ppb and the hydrogen content in the antibacterial gel is ≤1000 ppb. The content of the antibiotic in the antibacterial gel is 0.1-5 μg / mL, based on the total volume of the antibacterial gel. The gel carrier is poloxamer gel.

2. The antibacterial gel according to claim 1, wherein The hydrogen donor is a hydride.

3. The antibacterial gel according to claim 2, wherein The hydride is selected from lithium hydride (LiH), palladium hydride (PdH), beryllium hydride (BeH2), magnesium hydride (MgH2), calcium hydride (CaH2), strontium hydride (SrH2), titanium hydride (TiH2), aluminum hydride (AlH3), boron hydride (BH3), lithium borohydride (LiBH4), sodium borohydride (NaBH4), potassium borohydride (KBH4), magnesium borohydride (Mg(BH4)2), calcium borohydride (Ca(BH4)2), lithium aluminum hydride (LiAlH4), sodium aluminum hydride (NaAlH4), potassium aluminum hydride (KAlH4), magnesium aluminum hydride (Mg(AlH4)2), calcium aluminum hydride (Ca(AlH4)2), ammonia borane (NH3BH3), lithium amide (LiNH2), lithium imide (Li2NH) and a mixture thereof.

4. The antibacterial gel according to claim 2, wherein The hydride is lithium hydride, calcium hydride, palladium hydride, or magnesium hydride.

5. The antibacterial gel according to claim 4, wherein The hydride is magnesium hydride.

6. The antibacterial gel according to claim 2, wherein The hydride is a micro-nano hydride.

7. The antibacterial gel according to claim 6, wherein The hydride is micro-nano magnesium hydride.

8. The antibacterial gel according to claim 6 or 7, characterized in that The particle size of the micro-nano hydride is 0.1-100 μm.

9. The antibacterial gel according to claim 2, wherein The antibacterial gel comprises poloxamer gel and hydrogenated compounds, wherein the poloxamer gel and hydrogenated compounds are mixed together or exist separately.

10. The antibacterial gel according to claim 2, wherein The poloxamer is selected from the group consisting of poloxamer 407, poloxamer 188, poloxamer 184, or a combination thereof.

11. The antibacterial gel according to claim 2, wherein The weight percentage of the poloxamer is 15%-28% (w / v), based on the total volume of the antibacterial gel.

12. The antibacterial gel according to claim 10, wherein The content of the hydride is 0.2 mg / mL-5 mg / mL, based on the total volume of the antibacterial gel.

13. The antibacterial gel according to claim 2, wherein The antibacterial gel comprises the following components: 15%-28% (w / v) of poloxamer, 0.2mg / mL-3mg / mL of micro-nano magnesium hydride, and the balance of water or physiological saline, based on the total volume of the antibacterial gel.

14. Use of the antibacterial gel according to claim 1 for preparing a medicament for killing microorganisms.

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