WO2022144867A1 - Antimicrobial compounds based on glucoheptonic acids and their salts - Google Patents

Abstract

The invention relates to methods for disinfecting or sanitizing. The invention is also directed to methods for the preparation of certain compounds useful as a disinfectant or sanitizer, such as glucoheptonic acid, chlorhexidine diglucoheptonate, benzalkonium glucoheptonate, and combinations thereof. The invention is still further directed to methods for evaluating a compound for performance in microbial disinfecting or sanitizing.

Description

ANTIMICROBIAL COMPOUNDS BASED ON GLUCOHEPTONIC ACIDS AND THEIR SALTS

CROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to and the benefit of Provisional Application Serial No. 63/133,440, filed January 4, 2021, which is incorporated herein by reference in its entirety for all relevant purposes.

FIELD OF THE INVENTION

[0002] Provided herein are methods for disinfecting or sanitizing. Also provided herein are methods for the preparation of certain compounds useful as a disinfectant or sanitizer, such as glucoheptonic acid, chlorhexidine diglucoheptonate, benzalkonium glucoheptonate, and combinations thereof. Still further provided herein are methods for evaluating a compound for performance in microbial disinfecting or sanitizing.

BACKGROUND TO THE INVENTION

[0003] Chlorhexidine is a compound used in the medical field and in dentistry as an oral disinfectant. Benzalkonium chloride has been traditionally used in a variety of products, including laundry fabric softeners, shampoos, hair conditioners, and other personal care products. Benzalkonium chloride may also be found in a variety of pharmaceutical products such as eye, ear, or nasal drops. However, it is believed that certain microbes are beginning to develop resistance and/or tolerance to benzalkonium chloride.

[0004] There remains a need in the art to develop disinfecting or sanitizing compounds effective against a growing number of novel viruses, such as SARS-CoV-2. In connection with the growing number of novel viruses, there is also a need to develop new disinfecting or sanitizing compounds including more effective benzalkonium salts that reduce microbial populations effectively and are not prone to microbe resistance. Still further, there is a need to improve the process of making the feedstocks for preparing such new disinfecting or sanitizing compounds.

SUMMARY OF THE INVENTION

[0005] One aspect of the present invention is a method of disinfecting or sanitizing. The method comprises contacting an aqueous composition comprising a component selected from the group consisting of glucoheptonic acid, chlorhexidine diglucoheptonate, benzalkonium glucoheptonate, and combinations thereof with a microbial population. The Log Reduction is about 3 or greater, 30 seconds after application. [0006] Another aspect of the present invention is directed to a method of preparing glucoheptonic acid. The method comprises dissolving a glucoheptonate salt in water to form a solution; combining the solution with an acidic ion exchange resin to form a slurry; filtering the slurry to remove the ion exchange resin, and produce glucoheptonic acid. Optionally, the liquid from the filtered slurry may be dried to form the glucoheptonic acid. The conversion of the glucoheptonate salt to glucoheptonic acid is about 99% or greater, about 99.1% or greater, about 99.2% or greater, about 99.3% or greater, about 99.4% or greater, or about 99.5% or greater.

[0007] A further aspect of the present invention is directed to a method of preparing a benzalkonium glucoheptonate. The method comprises combining a benzalkonium salt and a glucoheptonate salt in a container and stirring the combination. Optionally, the combination is heated to a temperature of about 90° C or greater. Optionally, the combination is subjected to a nitrogen purge step during at least a portion of the heating.

[0008] An additional aspect of the present invention is directed to a method of evaluating a compound for performance in microbial disinfecting or sanitizing. The method comprises conducting a Bubble Pressure Tensiometry test of a compound; evaluating the reduction in surface tension exhibited by a compound as a function of time; and comparing the surface tension reduction to a compound known to exhibit microbial disinfecting or sanitizing properties to determine if the tested compound is useful for microbial disinfecting or sanitizing. [0009] Other objects and features will be in part apparent and in part pointed out hereinafter. BRIEF DESCRIPTION OF THE DRAWINGS

[0010] Figure 1 presents a comparison of the bubble age to surface tension for RM14A at various concentrations.

[0011] Figure 2 presents a comparison of the bubble age to surface tension for RM14B at various concentrations.

[0012] Figure 3 presents a comparison of the bubble age to surface tension for RM14C at various concentrations.

[0013] Figure 4 presents a comparison of the bubble age to surface tension for RM12A, RM 13 A, RM14A, and RM15A at super-CMC concentrations.

[0014] Figure 5 presents a comparison of the bubble age to surface tension for RM12A, RM 13 A, RM14A, and RM15A at CMC concentrations.

[0015] Figure 6 presents a comparison of the bubble age to surface tension for RM12A,

RM 13 A, RM14A, and RM15A at CMC concentrations. [0016] Figure 7 presents a comparison of bubble age to surface tension for RM12A, RM13A, RM14A, and RM15A at sub-CMC concentrations.

[0017] Figure 8 reports the conductivity for RM14A for the outer data points.

[0018] Figure 9 reports the conductivity for RM14A for the inner data points.

[0019] Figure 10 reports the conductivity for RM14A for the all data points.

[0020] Figure 11 reports the refractive index of RM12A, RM12B and RM12C.

[0021] Figure 12 reports the refractive index of RM13A, RM13B and RM13C.

[0022] Figure 13 reports the refractive index of RM14A, RM14B and RM14C.

[0023] Figure 14 reports the refractive index of RM15A.

[0024] Corresponding reference characters indicate corresponding parts throughout the drawings.

DETAILED DESCRIPTION OF THE INVENTION

[0025] In accordance with the present invention, methods for disinfecting or sanitizing have been devised that utilize disinfecting or sanitizing components previously not known to exhibit such properties and/or having surprising improvements over similar compounds. In particular, the methods of the present invention of disinfecting or sanitizing comprise contacting an aqueous composition comprising a component selected from the group consisting of glucoheptonic acid, chlorhexidine diglucoheptonate, benzalkonium glucoheptonate, and combinations thereof with a microbial population. Generally, contact with one or more of these components achieves a Log Reduction 30 seconds after application that is about 3 or greater.

[0026] Processes of the present invention are also directed to the use of diglucoheptonate salts having a certain concentration of a or [3 forms. Such processes include, for example, the use of an aqueous composition comprising chlorhexidine di-a-glucoheptonate and chlorhexidine di- P-glucoheptonate wherein the composition comprises a chlorhexidine di-a-glucoheptonate concentration, based on total isomers of chlorhexidine diglucoheptonate, of about 90% or greater. Alternatively, the microbial population is contacted with an aqueous composition comprising chlorhexidine di-a-glucoheptonate and chlorhexidine di-P-glucoheptonate wherein the composition comprises a chlorhexidine di-P-glucoheptonate concentration, based on total isomers of chlorhexidine diglucoheptonate, of about 25% or greater.

[0027] The present invention is further directed to methods of preparing glucoheptonic acid. The method provides improved conversion rates using a process that is less complex than previously used electrodialysis processes. The methods generally comprise dissolving a glucoheptonate salt in water to form a solution combining the solution with an acidic ion exchange resin to form a slurry; and filtering the ion exchange resin from the slurry to form glucoheptonic acid. Optionally, the liquid from the filtered slurry may be dried to form the glucoheptonic acid. The method achieves a conversion of the glucoheptonate salt to glucoheptonic acid of about 99% or greater, about 99.1% or greater, about 99.2% or greater, about 99.3% or greater, about 99.4% or greater, or about 99.5% or greater.

[0028] Still further, the present invention is directed to methods of preparing chlorhexidine diglucoheptonates using the glucoheptonic acid prepared in this manner. The process comprises combining chlorhexidine and water to form a slurry, adding the glucoheptonic acid to the slurry, and mixing the combination until the glucoheptonic acid is dissolved.

[0029] The present invention is also directed to methods of preparing a benzalkonium glucoheptonate. The method comprises combining a benzalkonium salt and a glucoheptonate salt in a container and stirring the combination. Optionally, the combination is heated to a temperature of about 90°C or greater. Optionally, the combination is subjected to a nitrogen purge step during at least a portion of the heating.

[0030] Further, the present invention is directed to methods of evaluating a compound for performance in microbial disinfecting or sanitizing. The method comprises conducting a Bubble Pressure Tensiometry test of a compound; evaluating the reduction in surface tension exhibited by a compound as a function of time; and comparing the surface tension reduction to a compound known to exhibit microbial disinfecting or sanitizing properties to determine if the tested compound is useful for microbial disinfecting or sanitizing.

[0031] In one embodiment of the present invention, a process for preparing a glucoheptonate salt generally comprises the reaction of a glucoheptonic acid with a free base in the reaction scheme set forth below. An example of such a reaction is the reaction of chlorhexidine with glucoheptonic acid to produce chlorhexidine diglucoheptonate.

B: + Glucoheptonic Acid - ► [BH] + [Glucoheptonate]"

[0032] Still further embodiments of the present invention are directed to reacting a glucoheptonate salt with a quaternary ammonium salt to form a desired benzalkonium glucoheptonate salt. For example, reaction of a benzalkonium salt with sodium glucoheptonate. [0033] Quaternary ammonium compounds (QAC's) are known to be effective as virucides. However, the growing preference in the global marketplace is to prepare at least a portion of such compounds from renewable resources in efforts to improve resource sustainability and reduce reliance on petrochemical feedstocks. One particular aspect of the present invention is directed to processes for preparing the quaternary ammonium salt, benzalkonium glucoheptonate from non-petrochemical components. For example, the benzalkonium component may be prepared from either coconut oil or palm kernel oil based amines, while the glucoheptonate portion of the molecule may be prepared from natural sugars that include glucose.

[0034] While particular quaternary ammonium compounds are discussed herein, it should be understood that the processes described herein are applicable to a wider variety of quaternary compounds, particularly where the quaternary ammonium components are paired with polyhydroxycarboxylic acids, using the free acid or salt form.

[0035] Still further aspects of the present invention are directed to use of the glucoheptonate salts or acids described herein for the destruction of certain undesirable microbes. For example, the destruction of yeast/fungi, viruses, bacteria, etc.

[0036] Glucoheptonate salts (e.g., sodium glucoheptonate) may be prepared from a variety of saccharide sources. For example, glucose, com syrup, molasses, molasses bottoms, black strap molasses, and combinations thereof. Generally, glucoheptonate is formed as the reaction product between a sugar and cyanide. For example, sodium cyanide.

[0037] In one embodiment, sodium glucoheptonate is formed by reacting glucose and sodium cyanide. Sodium cyanide is reacted with an approximately equimolar quantity of glucose at a temperature of 0-60°C, under mildly alkaline conditions over a several hour period. Ammonia is a byproduct of this reaction and is removed during the course of the reaction. When glucose is used to produce sodium glucoheptonate, the resulting aqueous product is composed of two glucoheptonates, in their a- and [3-isomeric forms. The general reaction scheme of glucose to sodium glucoheptonate is set forth below.

a-Isomer p-Isomer

(D-glycero-D-glucoheptonate) (D-glycero-D-idoheptonate)

[0038] In certain embodiments, the resulting mother liquor of the reaction is reduced in volume, cooled, and a lightly colored a-glucoheptonate salt (e.g., sodium a-glucoheptonate) is precipitated from the liquor. Through this precipitation process it is possible to form a solid having a purity of the a-isomer of greater than 99%. Following the precipitation step, the remaining liquor is a mixture of both a- and [3-isomer, having a majority [3-isomer product. This remaining liquor containing a mixture of both isomers is generally darker in appearance. Preparing Glucoheptonic Acid

[0039] After the saccharide has been converted to a glucoheptonate salt (e.g., sodium glucoheptonate), the salt may be converted to glucoheptonic acid. One method of the present invention is directed to preparing glucoheptonic acid using an ion exchange resin. Although discussed below in the context of a sodium glucoheptonate process, it is understood that similar processes may be employed with other glucoheptonate salts.

[0040] Previously known procedures for the conversion of a glucoheptonate salt to a glucoheptonic acid comprised the use of an electrodialysis system. However, use of such a process requires significant capital expenditure and processing costs. Further, such systems typically exhibit yields of about 98%. In contrast, the inventors have discovered that use of an ion exchange resin as described herein presents a simpler process for the conversion and results in conversion rates of up to 99% or greater. In the ion exchange process of the present invention, for sodium a-glucoheptonate samples, it was found by sodium ion analysis that the conversion was >99.3% effective. For sodium [3-glucoheptonate samples, it was found that the conversion was >99.1% effective. This additional conversion rate is significant when attempting to produce commercial scale quantities of the glucoheptonic acid.

[0041] In one process of the present invention, sodium glucoheptonate is contacted with an acidic ion exchange resin to convert the sodium salt to glucoheptonic acid. An acid ion exchange resin (as a water/resin slurry) is loaded into a flash chromatography column at ambient temperature. Sodium glucoheptonate is then dissolved in water, loaded into the column, and the column is eluted with water. The resulting eluate comprises a solution of glucoheptonic acid. Various iterations of this process may be conducted with different glucoheptonate salts. For example, in one embodiment, sodium a-glucoheptonate may be introduced into the column to produce a solution of a-glucoheptonic acid. In another embodiment, a sodium [373 -glucoheptonate solution may be introduced into the column to produce a [373 -glucoheptonic acid solution. The resulting glucoheptonic acid may be optionally dried in a forced air oven. The sodium [373 -glucoheptonate solution is a solution of sodium glucoheptonate comprising from about 60% to about 75% [3-isomer. For example, from about 61% to about 74%, from about 61% to about 73%, from about 61% to about 72%, from about 61% to about 71%, from about 62% to about 71%, from about 63% to about 71%, from about 64% to about 71%, from about 65% to about 71%, from about 66% to about 71%, from about 67% to about 71%, from about 68% to about 71%, or from about 69% to about 71% of the [3- isomer. In certain embodiments, the sodium [373-glucoheptonate solution comprises about 67- 71% [3-isomer and about 29-33% a isomer. In another embodiment, the sodium [373- glucoheptonate solution comprises about 61-73.5% [3-isomer and about 26.5-39% a isomer. In some embodiments, the sodium [373-glucoheptonate solution is a solution having a typical ratio of a to [3 isomer, having a relative absence of borate, and having a low solids content. For example, in some embodiments, the sodium [373-glucoheptonate solution has a solids content at room temperature of about 20 wt.% or less, about 15 wt.% or less, about 10 wt.% or less, about 5 wt.% or less, or about 1 wt.% or less.

[0042] In another process of the present invention, sodium glucoheptonate is dissolved in water in a first container. An acidic ion exchange resin is then added to the first container to form a slurry. The slurry is mixed by pouring the combination between a first and a second container (e.g. from the first container to a second container, from the second container to the first container, etc.) at 20 minute intervals for a period of four hours. The resulting mixed slurry is then filtered to remove the ion exchange resin and yield a solution of glucoheptonic acid. The process of pouring the combination between a first and a second container may be untaken at a variety of intervals. For example, at 5 minute intervals for a period of at least about 30 minutes, 5 minute intervals for a period of at least about 1 hour, 10 minute intervals for a period of at least about 1 hour, 10 minute intervals for a period of at least about 2 hours, 10 minute intervals for a period of at least about 3 hours, 10 minute intervals for a period of at least about 4 hours, 15 minute intervals for a period of at least about 4 hours, 20 minute intervals for a period of at least about 4 hours, or 20 minute intervals for a period of at least about 5 hours. [0043] This process may be conducted with various glucoheptonate salts. For example, in one embodiment, sodium a-glucoheptonate (i.e. a solution comprising less than 1% of the [3 isomer of sodium glucoheptonate) is combined with the ion exchange resin to produce a slurry of a- glucoheptonic acid after mixing between a first and a second container as described above. The resulting a-glucoheptonic acid is optionally dried in a forced air oven (e.g., at a temperature of about 65 °C) to form a film. After drying, in certain embodiments, the a-glucoheptonic acid concentration based on total isomers is about 90% or greater, as determined by HPLC. In other embodiments, the a-glucoheptonic acid concentration based on total isomers is about 95% or greater, about 96% or greater, about 97% or greater, about 98% or greater, about 99% or greater, about 99.5% or greater, or about 99.9% or greater.

[0044] In another embodiment, a sodium [373 -glucoheptonate solution is combined with an ion exchange resin to produce a slurry of [373-glucoheptonic acid after mixing between a first and a second container as described above. The resulting glucoheptonic acid is optionally dried in a forced air oven (e.g., at a temperature of about 65 °C) to form a film. After drying, in certain embodiments, the ratio of [3-glucoheptonic acid to a-glucoheptonic acid may be 2.61 (73.2% [3 isomer) as determined by HPLC. In other embodiments, the ratio of [3-glucoheptonic acid to a- glucoheptonic acid may be about 1.5 or greater, about 2.0 or greater, about 2.5 or greater, about 3.0 or greater, about 3.5 or greater about 4.0 or greater, or about 4.5 or greater.

[0045] The ion exchange resin used in the process of the present invention to convert a glucoheptonate salt to glucoheptonic acid may be any resin suitable for such a conversion. In certain embodiments, the ion exchange resin is an acidic resin. For example, a strong acid cation type resin. In one embodiments, the strong acid cation exchange resin has a sulfonic acid functional group. Without being bound by the theory, it is believed that the presence of a strong acid functional group aids in the more complete conversion of a glucoheptonate salt to glucoheptonic acid. In certain other embodiments, the ion exchange resin comprises a styrenedivinyl benzene matrix.

[0046] In various embodiments, the ion exchange resin has a degree of crosslinking from about 1% to about 25%. For example, from about 2% to about 25%, from about 2% to about 20%, from about 2% to about 19%, from about 2% to about 18%, from about 2% to about 17%, from about 2% to about 16%, from about 4% to about 16%, from about 4% to about 14%, from about 4% to about 12%, from about 4% to about 10%, from about 4% to about 9%, from about 5% to about 9%, from about 6% to about 9%, or from about 7% to about 9%. In some embodiments, the ion exchange resin has a degree of crosslinking of about 1% or greater, about 2% or greater, about 3% or greater, about 4% or greater, or about 5% or greater.

[0047] In certain embodiments, the ion exchange resin is macroporous.

[0048] In some embodiments the ion exchange resin has a mean particle size of from about 300 pm to about 700 pm, from about 350 pm to about 700 pm, from about 375 pm to about 700 pm, from about 400 pm to about 700 pm, from about 400 pm to about 675 pm, from about 400 pm to about 650 pm, from about 400 pm to about 625 pm, from about 400 pm to about 600 pm, from about 425 pm to about 600 pm, from about 450 pm to about 600 pm, from about 475 pm to about 600 pm, from about 500 pm to about 600 pm, from about 525 pm to about 600 pm, or from about 550 pm to about 600 pm. In certain other embodiments, the ion exchange resin has a mean particle size of from about 400 pm to about 900 pm, from about 450 pm to about 900 pm, from about 500 pm to about 900 pm, from about 550 pm to about 900 pm, from about 600 pm to about 900 pm, from about 600 pm to about 850 pm, from about 600 pm to about 800 pm, from about 600 pm to about 750 pm, or from about 650 pm to about 700 pm.

[0049] In various embodiments, the ion exchange resin comprises monodisperse ion exchange particles (e.g., beads).

[0050] In one embodiment, the ion exchange resin is a DOWEX MARATHON MSC series cation exchange resin. In another embodiment, the ion exchange resin is a LEWATIT MONOPLUS SP 112 H series cation exchange resin.

Preparing a Chlorhexidine Salt

[0051] In processes of the present invention, the glucoheptonic acid may be reacted with a base in order to prepare the desired glucoheptonate compound.

[0052] In certain processes of the present invention, glucoheptonic acid is reacted with chlorhexidine in order to prepare the desired chlorhexidine glucoheptonate compound. [0053] Other aspects of the present invention are directed to the formation and use of chlorhexidine diglucoheptonate salts. As set forth in the general reaction scheme below, chlorhexidine is reacted with glucoheptonic acid to form chlorhexidine diglucoheptonate.

[0054] In certain embodiments, the chlorhexidine diglucoheptonate of the present invention is prepared be reacting a glucoheptonic acid with chlorhexidine, wherein the process comprises a molar excess of the glucoheptonic acid. In certain embodiments, the chlorhexidine diglucoheptonate of the present invention has a molar ratio of glucoheptonic acid to chlorhexidine of from about 1.5: 1 to about 3: 1, from about 2: 1 to about 2.9: 1, from about 2: 1 to about 2.8: 1, from about 2: 1 to about 2.7: 1, from about 2: 1 to about 2.6: 1, or from about 2.1 : 1 to about 2.6: 1. In other embodiments, the chlorhexidine diglucoheptonate of the present invention has a molar ratio of glucoheptonic acid to chlorhexidine of from about 1 : 1 to about 5: 1, from about 1: 1 to about 4: 1, from about 1: 1 to about 3: 1, from about 1: 1 to about 2.5: 1, from about 1: 1 to about 2.0: 1, from about 1: 1 to about 1.9: 1, from about 1: 1 to about 1.8: 1, from about 1: 1 to about 1.7: 1, from about 1: 1 to about 1.6: 1, or from about 1: 1 to about 1.5: 1. Preparing a Benzalkonium Salt

[0055] In other embodiments, it may be more economical to utilize a counterion in the form of a salt, instead of a base. For example, unlike the free base chlorhexidine, benzalkonium chloride is a salt and presents difficulties in preparing alternate salts. Thus, where it is desired to form a benzalkonium glucoheptonate, it is preferred to react a benzalkonium salt with a glucoheptonate salt. For example, the benzalkonium salt may be benzalkonium chloride and the glucoheptonate salt may be sodium glucoheptonate.

[0056] The benzalkonium chloride used in exemplary embodiments of the present invention is generally formed by the reaction scheme set forth below, wherein R is 40% C12, 50% C14, and 10% Ci6.

[0057] Generally, the reaction of a benzalkonium salt with a glucoheptonate salt comprises combining the two compounds in a container and mixing. Optionally, the temperature of the combination may be increased (e.g,. to about 90°C). Optionally, the process may further comprise the use of a nitrogen purge to remove excess moisture.

[0058] One challenge presented by the proposed process of reacting a benzalkonium salt with a glucoheptonate salt is that the reaction yields a stoichiometric amount of a metal salt. Since the virucidal action of benzalkonium glucoheptonate relies on formation of an ion pair between the polyhydroxycarboxylate anion and the quaternary ammonium cation, the additional equivalent of metal salt must be removed. Therefore, it may be required to conduct one or more of the following process steps prior to, during, or after the reaction of the benzalkonium salt (benzalkonium chloride) with the glucoheptonate salt:

1. Ion exchange purification of one or both of the benzalkonium and glucoheptonic streams prior to mixing (aqueous).

2. Mixing the benzalkonium and glucoheptonic streams without a solvent, and then subsequently drying the medium, with or without vacuum.

3. Mixing the benzalkonium and glucoheptonic streams in a suitable solvent, and subsequently drying the mixture.

4. Centrifugation, precipitation, and/or filtration.

5. Solvent removal .

6. Drying. 7. Concentration of the benzalkonium glucoheptonate salt by removal of liquid through heating.

8. Dialysis.

9. Electrodialysis.

10. Ion exchange purification of the benzalkonium glucoheptonate salt product. [0059] For example, in one embodiment of the present invention, benzalkonium glucoheptonate is formed by the reaction scheme below, utilizing one or more of the optional processing steps set forth above.

NaCl

[0060] Any suitable solvent useful for forming benzalkonium glucoheptonate may be used in the above reaction. For example, the solvent may be selected from the group consisting of methanol, ethanol, isopropyl alcohol, n-propyl alcohol, acetone, butyl alcohol, and combinations thereof. Butyl alcohol should be understood to include one or more of n-butanol, sec-butanol, isobutanol and tert-butanol.

[0061] In certain embodiments, the benzalkonium glucoheptonate of the present invention is prepared be reacting a benzalkonium salt with a glucoheptonate salt, wherein the process comprises a molar excess of the glucoheptonate salt. For example, in one embodiment, the molar ratio of glucoheptonate salt to benzalkonium salt is from about 1: 1 to about 5: 1, from about 1 : 1 to about 4: 1, from about 1 : 1 to about 3: 1, from about 1 : 1 to about 2.5: 1, from about 1 : 1 to about 2.0: 1, from about 1 : 1 to about 1.9: 1, from about 1 : 1 to about 1.8: 1, from about 1 : 1 to about 1.7: 1, from about 1: 1 to about 1.6: 1, or from about 1: 1 to about 1.5: 1. In certain other embodiments, the molar ratio of glucoheptonate salt to benzalkonium salt is from about 1.1 : 1 to about 5: 1, from about 1.1: 1 to about 4: 1, from about 1.1: 1 to about 3: 1, from about 1.1 : Ito about 2.5: 1, from about 1.1: Ito about 2.0: 1, from about 1.1: 1 to about 1.9: 1, from about 1.1: 1 to about 1.8: 1, from about 1.1: 1 to about 1.7: 1, from about 1.1: 1 to about 1.6: 1, or from about 1.1: 1 to about 1.5: 1.

[0062] While the specific example of benzalkonium glucoheptonate has been provided herein, there are many other compounds that may be prepared with similar expectations of efficacy. The benzalkonium counterion described herein is an example of a broader class of cationics. Therefore, other suitable sources of cationic counterions may include, but are not necessarily limited to, benzethonium chloride, polymeric quaternary ammonium salts, dialkyldimethyl quaternary ammonium salts, dialkylmethyl ammonium quaternary ammonium salts with twin tails, other alkyl or hydroxyalkyl substituted quaternary ammonium salts, cetylpyridinium chloride, benzyl substituted quaternary ammonium salts, picolinium salts, imidazolinium salts, N-ethyl-Morpholinium salts, isoquinolinium salts, chlorhexidine salts, and combinations thereof. The aforementioned sources of cationic counterions may be used to prepare glucoheptonate compounds that are expected to exhibit similar efficacy, for example by reacting the aforementioned cationic counterions with a glucoheptonate salt.

[0063] For example, the glucoheptonate compound may be selected from the group consisting of benzethonium glucoheptonate, polymeric quaternary ammonium glucoheptonates, and cetylpyridinium glucoheptonate.

[0064] In certain embodiments, the glucoheptonate compound may be a quaternary ammonium salt having the formula: RIR2R3R4N⁺X", wherein Ri and R2 are independently selected from the group consisting of straight, branched, or cyclic alkyl or alkenyl groups having from about 8 to about 18 carbon atoms, and R3 and R4 are independently selected from the group consisting of straight alkyl groups having from about Ito about 4 carbon atoms, hydroxyethyl, hydroxypropyl, hydroxybutyl, -(CFbCFbC mCFbCFbOH, (CH2CHCH3 O)_mCH₂CHCH₃ OH,

(CH2CH₂O)_m(CH2CHCH3O)_nCH2CHCH3OH, or -(CH2CHCH3O)_m(CH2CH2O)nCH2CH2OH, wherein m is an integer from about 1 to about 10 and n is an integer from about 1 to about 10, and X" is glucoheptonate.

[0065] In certain embodiments, the glucoheptonate compound may be a quaternary ammonium salt having the formula: R1 R2R R4N 'X-. wherein Ri is selected from the group consisting of 3 -alkoxy-2 -hydroxypropyl ora straight, branched, or cyclic alkyl or alkenyl group having from about 8 to about 18 carbon atoms, R2, R3 and R> are independently selected from the group consisting of straight alkyl groups having from about 1 to about 4 carbon atoms, hydroxyethyl, hydroxypropyl, hydroxybutyl, -(CH2CH2O)_mCH2CH2OH, (CH₂CHCH3O)_mCH2CHCH₃OH, -(CH2CH2O)_m(CH2CHCH3O)nCH2CHCH3OH, or - (CH2CHCH3O)m(CH2CH2O)nCH2CH₂OH, wherein m is an integer from about 1 to about 10 and n is an integer from about 1 to about 10, and X" is glucoheptonate.

[0066] In another embodiment, the glucoheptonate compound may be a quaternary ammonium salt having the formula: R1 R2R R4N 'X-. wherein Ri is selected from the group consisting of a straight, branched, or cyclic alkyl or alkenyl group having from about 8 to about 18 carbon atoms, R2 is selected from the group consisting of a substituted or unsubstituted benzyl, ethylbenzyl, naphthyl, or methylnaphthyl, and R3 and R4 are independently selected from the group consisting of straight alkyl groups having from about 1 to about 4 carbon atoms, hydroxyethyl, hydroxypropyl, hydroxybutyl, -(CH2CH2O)_mCH2CH2OH, (CH2CHCH3O)mCH₂CHCH3OH, -(CH2CH2O)m(CH2CHCH3O)nCH₂CHCH3OH, or (CH2CHCH₃O)m(CH2CH2O)_nCH2CH2OH, wherein m is an integer from about 1 to about 10 and n is an integer from about Ito about 10, and X" is glucoheptonate.

[0067] In yet another embodiment, the glucoheptonate compound may be a quaternary ammonium salt having the formula RIR2R R4N⁺X", wherein Ri is selected from the group consisting of 3 -alkoxy-2 -hydroxypropyl ora straight, branched, or cyclic alkyl or alkenyl group having from about 8 to about 18 carbon atoms, R2 is selected from the group consisting of a substituted or unsubstituted benzyl, ethylbenzyl, naphthyl, or methylnaphthyl, a straight, branched, or cyclic alkyl or alkenyl group having from about 8 to about 18 carbon atoms, a straight alkyl group having from about 1 to about 4 carbon atoms, hydroxyethyl, hydroxypropyl, hydroxybutyl, -(CH2CH2O)_mCH2CH2OH, -(CH2CHCH3O)_mCH2CHCH3OH, -(CH2CH2O)_m(CH₂CHCH3 O)_nCH₂CHCH₃ OH, or -(CH2CHCH3 O)_m(CH2CH₂O)_nCH2CH2OH, wherein m is an integer from about 1 to about 10, n is an integer from about 1 to about 10, R3 and R4 are independently selected from the group consisting of a straight alkyl group having from about 1 to about 4 carbon atoms, hydroxyethyl, hydroxypropyl, hydroxybutyl, (CH2CH2O)_mCH2CH₂OH, -(CHzCHCftO^CHzCHOHOH,

-(CH2CH2O)_m(CH₂CHCH3 O)_nCH₂CHCH₃ OH, or -(CH2CHCH3 O)_m(CH2CH₂O)_nCH2CH2OH, where m is an integer from about 1 to about 10 and n is an integer from about 1 to aboutlO, and X" is glucoheptonate. [0068] For example, in some embodiments, the glucoheptonate compound may be selected from the following, wherein X" is glucoheptonate:

Applications of Glucoheptonate Compounds

[0069] The glucoheptonate compounds disclosed herein may be useful as antimicrobials or virucides. Such glucoheptonate compounds include, for example, glucoheptonic acid, chlorhexidine diglucoheptonate, benzalkonium glucoheptonate, and combinations thereof. The glucoheptonate compounds may be present in a composition in an amount sufficient to achieve the desired antimicrobial or virucidal effects. For example, it may be desirable to form a composition having a certain minimum inhibitory concentration (MIC), defined as the lowest concentration of an antimicrobial that will inhibit the visible growth of a microorganism after overnight incubation. In other embodiments, it may be desirable to form a composition having a certain minimum bactericidal concentration (MBC), defined as the lowest concentration of antimicrobial that will prevent the growth of an organism. In still other embodiments, it may be desirable to form a composition having a concentration suitable for achieving a certain Log Reduction (e.g., Log 3 Reduction after 30 seconds).

[0070] In certain embodiments, glucoheptonic acid is present in a composition comprising from about 0.5 to about 10 wt%, from about 0.5 to about 9 wt%, from about 0.5 to about 8 wt%, from about 0.5 to about 7 wt%, from about 0.5 to about 6 wt%, from about 0.6 to about 6 wt%, from about 0.7 to about 6 wt%, from about 0.8 to about 6 wt%, from about 0.9 to about 6 wt%, from about 1 to about 6 wt%, from about 2 to about 6 wt%, from about 3 to about 6 wt%, from about 4 to about 6 wt%, or from about 5 to about 6 wt% of glucoheptonic acid. [0071] In one embodiment, chlorhexidine diglucoheptonate is present in a composition comprising from about 0.005 to about 10 wt%, from about 0.005 to about 9 wt%, from about 0.005 to about 8 wt%, from about 0.005 to about 7 wt%, from about 0.005 to about 6 wt%, from about 0.006 to about 6 wt%, from about 0.007 to about 6 wt%, from about 0.008 to about

6 wt%, from about 0.009 to about 6 wt%, from about 0.01 to about 6 wt%, from about 0.011 to about 6 wt%, from about 0.012 to about 6 wt%, from about 0.013 to about 6 wt%, from about 0.014 to about 6 wt%, from about 0.015 to about 6 wt%, or from about 0.015 to about 5 wt% of chlorhexidine diglucoheptonate.

[0072] In some embodiments, benzalkonium glucoheptonate is present in a composition comprising from about 0.00025 to about 10 wt%, from about 0.0005 to about 10 wt%, from about 0.0005 to about 9 wt%, from about 0.0005 to about 8 wt%, from about 0.0005 to about

7 wt%, from about 0.0005 to about 6 wt%, from about 0.0005 to about 5 wt%, from about 0.0005 to about 4 wt%, from about 0.001 to about 4 wt%, from about 0.002 to about 4 wt%, from about 0.003 to about 4 wt%, from about 0.004 to about 4 wt%, from about 0.005 to about 4 wt%, from about 0.01 to about 4 wt%, from about 0.02 to about 4 wt%, from about 0.04 to about 4 wt%, from about 0.06 to about 4 wt%, from about 0.08 to about 4 wt%, or from about 0.10 to about 4 wt% of benzalkonium glucoheptonate.

[0073] Preparing the glucoheptonate compound from raw materials such as those described above (e.g., glucose) allows for the production of an antimicrobial compound having a desirable renewable carbon index. The glucoheptonate compounds have, for example, a renewable carbon index of about 50% or greater, about 55% or greater, about 60% or greater, about 65% or greater, about 70% or greater, about 75% or greater, about 80% or greater, about 85% or greater, about 90% or greater, or about 95% or greater.

[0074] The specific action of the antimicrobial is highly dependent on its structure. Without being bound by the theory, it is believed that the glucoheptonate moiety of the compounds of the present invention attaches itself to the outer membrane(s) of the virus by hydrogen bonding, potentially allowing for a “Trojan Horse” delivery mechanism of the quaternary ammonium or free base portion. It is further believed that the seven carbon glucoheptonate chain may offer improved efficacy over other polyhydroxycarboxylic chains such as the six carbon gluconate chain.

[0075] Glucoheptonate compounds of the present invention may also be useful as algaecides, bactericides, tuberculocides, sporicides, and/or fungicides. The affinity of glucoheptonates for calcium and other polyvalent cations may also enhance the antimicrobial efficacy of the compounds, especially when cell membranes of the target microbe contain calcium or other critical polyvalent ions in the outer layers.

[0076] Without being bound to the theory, it is believed that counterions based on polyhydrocarboxylic acids or sugar acids (e.g., glucoheptonates) are useful against Gramnegative and Gram-positive bacterial cells because these counterions have similar structure to elements of the outer membrane of the bacteria cells. Gram-negative microbes have outer structures that are composed of lipopolysaccharides, which are long chain monosaccharide units joined by glycosidic bonds. Gram-positive bacteria are composed of peptidoglycan, which is comprised of alternating polymeric amine sugars. Similarly, enveloped viruses often contain glycoproteins, which contain oligosaccharide chains. It is believed that hydrogen bonding occurs between the oligosaccharide structures and sugar acid counterions, enhancing the destructive effect of the cationic portion of the molecule. It is further believed that the associations between the cell structures and the counterion can lead to "Trojan Horse" mechanisms of cell destruction referenced above.

[0077] Although reference is made herein to specific pathogens or microbes that may be reduced or destroyed by the compounds of the present invention, it should be understood that this discussion is not limiting. As explained in Sattar, Syed, Journal of AOAC International, Vol. 90, No. 6, 2007, it is generally recognized in the field that there is a hierarchy of susceptibility of viruses to chemical disinfectants. For example, enveloped viruses are generally highly susceptible to chemical disinfectants. It is reasonably expected that a chemical disinfectant determined to be useful for a particular virus will also be useful for viruses having the same or higher levels of susceptibility to chemical disinfectants.

[0078] In certain embodiments, the compounds of the present invention (e.g., chlorhexidine diglucoheptonate, benzalkonium glucoheptonate, glucoheptonic acid, etc.) are effective at the reduction or destruction of certain undesirable microbes. The compounds of the present invention may be useful for the destruction of yeast/fungi, viruses, bacteria, etc.

[0079] For example, the compounds of the present invention may be useful for the reduction or destruction of one or more yeast/fungi selected from the group consisting of Aspergillus niger, Candida albicans, Candida tropicalis, Rhodotorula mucilaginosa, and Penicillium species.

[0080] The compounds of the present invention may be useful for the reduction or destruction of one or more species selected from the group consisting of Mastadenovirus and Aviadenovirus (i.e. adenovirus), Norovirus (e.g., Norwalk Virus)' , Betacoronavirus (e.g., Middle East respiratory syndrome-related coronavirus [MERS-CoV] and Severe acute respiratory syndrome coronavirus [e.g., SARS-CoV-2]), Ebolavirus (e.g., Zaire ebolavirus)' , Flavivirus (e.g., West Nile virus and Zika virus), Orthohantavirus (i.e. hantavirus), Varicellovirus (e.g., Equid alphaherpesvirus species and Human alphaherpesvirus 3 [i.e. Varicella-zoster virus ), Rhadinovirus (e.g., Equid gammaherpesvirus species and Human gammaherpesvirus 8), Simplexvirus (e.g., Human alphaherpesvirus species), Lymphocryptovirus (e.g., Human gammaherpesvirus 4 [i.e. Epstein-Barr virus]), Cytomegalovirus (e.g., Human betaherpesvirus 5), Roseolovirus (e.g., Human betaherpesvirus species), Rubivirus (e.g., Rubivirus rubella [i.e. rubella]), Orthonairovirus (e.g., Crimean- Congo hemorrhagic fever orthonairovirus), Alphainfluenzavirus (i.e. Influenza A virus), Betainfluenzavirus (i.e. Influenza B virus), Morbillivirus (e.g., Measles morbillivirus and Canine morbillivirus [i.e. Canine distemper virus]), Respirovirus (e.g., Human respirovirus 1 and 3 [i.e. Human parainfluenza virus 1 and o |). Orthorubulavirus (e.g., Human orthorubulavirus 2 and 4 [i.e. Human parainfluenza virus 2 and 4], Mammalian orthorubulavirus 5 [i.e. Canine parainfluenza virus 5], Mumps orthorubulavirus [i.e. mumps]), Protoparvovirus (e.g., Carnivore protoparvovirus 1 [i.e. canine parvovirus and feline panleukopenia virus]), Enterovirus (e.g., Enterovirus C [i.e. Poliovirus or polio]), Orthopneumovirus (e.g., Human orthopneumovirus [i.e. Human respiratory syncytial virus, hRSV]), Orthopoxvirus (e.g., Variola virus [i.e. smallpox]), Rotavirus (e.g., bovine rotavirus and human rotavirus), Lentivirus (e.g., Human immunodeficiency virus 1 and 2 [i.e. HIV]), Lyssavirus (e.g., Rabies lyssavirus [i.e. rabies]), and combinations thereof.

[0081] In addition, the compounds of the present invention may be useful for the reduction or destruction of the various species commonly known as echovirus (e.g., of the genera Enterovirus, Orthoreovirus, and Parechovirus), as well as hepatitis A-F (e.g., of the genera Hepatovirus, Orthohepadnavirus, Hepacivirus, Deltavirus, Orthohepevirus, and Alphavirus).

[0082] The compounds of the present invention may be useful for the reduction or destruction of one or more bacteria selected from the group consisting of Acinetobacter baumannii, Bordatella pertussis, Enterobacter aerogenes, Enterobacter cloacae, Enterococcus faecalis (VRE), Enterococcus faecium, Bordatella pertussis, Corynebacterium xerosis, Escherichia coli, Klebsiella pneumoniae, Legionella pneumophila, Listeria monocytogenes, Micrococcus luteus, Mycobacterium terrae, Propionibacterium acnes, Staphylococcus aureus, methicillin- resistant Staphylococcus aureus (MRSA), Staphylococcus epidermidis, Streptococcus pneumoniae, Streptococcus pyogenes, Streptococcus sobrinus, Salmonella enterica, Salmonella abony, Pseudomonas aerginosa, Pseudomonas maltophilia, and combinations thereof.

[0083] As detailed in the examples, Bubble Pressure Tensiometry may be used to evaluate compounds of the present invention (i.e. surface active antimicrobials) for performance in microbial testing. Bubble Pressure Tensiometry differs from other equilibrium (thermodynamic) surface tension measurement techniques in that it provides dynamic (kinetic) information, and the impact of time on the rate and magnitude of surface tension reduction can be observed. Both of these are important parameters in any process that involves surfactants and surfaces, and elucidation of such properties may allow for pre-selection of active biocides. Without being bound by the theory, it is believed that the results of a bubble pressure tensiometry test of a compound may be a useful tool for pre-screening of compounds to determine the relative usefulness as a disinfectant

[0084] Additionally, without being bound by the theory, the structural differences between gluconate, a-glucoheptonate, and [3-glucoheptonate counterions are thought to allow for chlorhexidine salts of the [3-isomer to interact more effectively with a microbial membrane. EXAMPLES

Example 1 : Preparing q-Glucoheptonic Acid

[0085] An experiment was conducted to prepare a solution of a-glucoheptonic acid.

[0086] 50 g of an acidic ion exchange resin, as a water/resin slurry, was loaded into a flash chromatography column at ambient temperature. 15 g of sodium a-glucoheptonate was dissolved in 50 g water and then loaded into the column. The column was then eluted with water using five passes of 50 mb each. The resulting eluates were combined to form the solution of a-glucoheptonic acid. The solution was a clear, light straw-colored liquid. The pH of the solution was 2.32, and the solution had a solids content of 8.8 wt.%.

Example 2: Preparing q-Glucoheptonic Acid

[0087] A further experiment was conducted to prepare a solution of a-glucoheptonic acid. [0088] 150 g of sodium a-glucoheptonate was dissolved in 750 g of water in a first container. 500 g of an acidic ion exchange resin was placed in a second container. The acidic ion exchange resins DOWEX MARATHON MSC and LEWATIT MONOPLUS SP 112 H were tested in this experiment. The liquid from the first container was poured into the second container containing the resin, and the slurry was mixed by pouring one container into the other every 20 minutes for one hour or four hours. The slurry was then filtered to remove the ion exchange resin beads and yield a solution of a-glucoheptonic acid. The solution of a-glucoheptonic acid was a clear, light straw-colored liquid. The solution had a pH of approximately 2.30 and an average solids content of 10 wt.%. The results of this experiment are set forth below in Table 1. "65°C Solids" represents the weight percentage of solids present in the solution of a- glucoheptonic acid, where the % solids is determined by oven drying at 65°C. The sodium value represents the results of residual sodium quantification, evaluated by Inductively Coupled Plasma (ICP) analysis. The data was collected with a Spectro Blue-EOP-TI instrument using a Lowest Calibration Level (LCL) of 0.25 mg/kg.

Table 1

[0089] A portion of the a-glucoheptonic acid was dried at 65 °C in a forced air oven on a flat glass plate to generate a brittle white film. NMR and IR spectra analysis were characteristic of a polyhydroxycarboxylic acid compound in its acidic form. The a-glucoheptonic acid content was determined to be >99% of total isomers by HPLC.

[0090] The film material was also heated to 850°C in a muffle furnace to determine the ash content. The ash content was determined to be 1000 ppm.

Example 3: Preparing B73-Glucoheptonic Acid

[0091] An experiment was conducted to prepare a solution of [373-glucoheptonic acid.

[0092] 50 g of an acidic ion exchange resin, as a water/resin slurry, was loaded into a flash chromatography column at ambient temperature. 33.63 g of a sodium [373 -glucoheptonate solution was loaded into the column. The column was then eluted with water and 161 g of a [373-glucoheptonic acid solution was collected. The [373-glucoheptonic acid solution was a clear, dark straw-colored liquid.

Example 4: Preparing B73-Glucoheptonic Acid

[0093] A further experiment was conducted to prepare a solution of 73-glucoheptonic acid. [0094] 336.3 g of a sodium [373 -glucoheptonate solution was dissolved in 564 g of water in a first container. 500 g of an acidic ion exchange resin was placed in a second container. The liquid from the first container was poured into the second container containing the resin, and the slurry was mixed by pouring one container into the other every 20 minutes for four hours. The slurry was then filtered to remove the beads and yield a solution of solution of P73- glucoheptonic acid. The solution of p73-glucoheptonic acid was a clear, dark straw-colored liquid. The solution had a pH of approximately 2.56 and a solids content of approximately 11.4 wt.%. The results of this experiment are set forth below in Table 2. "65°C Solids" represents the weight percentage of solids present in the solution of p73-glucoheptonic acid, where the % solids is determined by oven drying at 65°C. The sodium value represents the results of residual sodium quantification, evaluated by Inductively Coupled Plasma (ICP) analysis. The data was collected with a Spectro Blue-EOP-TI instrument using a Lowest Calibration Level (LCL) of 0.25 mg/kg.

Table 2

[0095] A portion of the p73-glucoheptonic acid was dried at 65 °C in a forced air oven on a flat glass plate to generate a brittle white film. NMR and IR spectra analysis were characteristic of a polyhydroxycarboxylic acid compound in its acidic form. The ratio of P-glucoheptonic acid to a-glucoheptonic acid was determined to be 2.73 by HPLC (73.2% p isomer). The film material was heated to 850°C in a muffle furnace to determine the ash content. This procedure yielded no recoverable ash.

Example 5: Preparing P81-Glucoheptonic Acid

[0096] An experiment was conducted to prepare a solution of P81-glucoheptonic acid.

[0097] 476 g of a liquid comprising sodium glucoheptonate having a P-isomer content of 81% (referred to herein as sodium P81 -glucoheptonate) was added to a container and mixed with 120 mL of methanol. In certain embodiments, the sodium P81 -glucoheptonate may be prepared in the manner set forth in US 3,084,188. The container was placed in a refrigerator (2-8°C) and allowed to rest overnight. 20% of the volume of the container comprised crystals after resting overnight. The supernatant was dried at 65°C in a forced air oven on a flat glass plate for a period of two days and generated a solid. 10.57 g of the solid was removed from the glass plate, dissolved in 54 g of water, and combined with 36.3 g of an acidic ion exchange resin. The slurry was stirred every 20 minutes for a period of four hours. At the conclusion of this time period, the slurry was filtered to remove the ion exchange resin. The resulting solution of P81- glucoheptonic acid was a clear, dark straw-colored liquid with a pH of 1.93 and a solids content of 11.0 wt.%.

Example 6: Preparing Benzalkonium q-Glucoheptonate

[0098] An experiment was conducted to prepare benzalkonium a-glucoheptonate.

[0099] 1000 g of benzalkonium chloride was added to a round bottom flask equipped with overhead stirrer assembly. 545 g of sodium a-glucoheptonate was then added to the flask and the flask was mixed for 45 minutes. The temperature was increased from 30°C to 90°C and maintained at about 90°C for approximately 40 hours. During the final 20 hours, a slow nitrogen purge was maintained to remove excess moisture. The target moisture content was less than 1%. The result of this reaction was a brown paste having a pH of 6.5 (5% aq.). The theoretical yield was determined to be >99% benzalkonium a-glucoheptonate.

[0100] Titration with silver nitrate was undertaken and it was determined that the resulting material comprised 10.6 wt% sodium chloride.

Example 7: Testing Glucoheptonate Salts

[0101] Solutions of chlorhexidine di-a-glucoheptonate (RM13), chlorhexidine di-[373- glucoheptonate (RM14), and chlorhexidine di-|381 -glucoheptonate (RM15) were prepared for further testing. Chlorhexidine digluconate (RM12) was also prepared.

[0102] For each solution, chlorhexidine was slurried with water, the respective acid (gluconic acid, a-glucoheptonic acid, [373-glucoheptonic acid, or [381-glucoheptonic acid) was mixed until dissolved, and the mixture was diluted with water. The resulting solutions were 0. 10 molar solutions with respect to the chlorhexidine concentration. Molar ratios of the glucoheptonic or gluconic acid to chlorhexidine varied from 2.0: 1 to 2.6: 1.

[0103] Multiple solutions were prepared utilizing differing molar ratios of the chlorhexidine and the respective acid (gluconic acid, a-glucoheptonic acid, [373-glucoheptonic acid, or [381-glucoheptonic acid). Table 3 below sets forth the theoretical masses required for preparing a 0. 10 M salt solution based on the noted molar ratio.

Table 3

Example 8: Bubble Tensionmetry

[0104] Dynamic surface tensions of certain formulations were measured using a KRUSS BP100 BPT Mobile bubble pressure tensiometer. Samples of the solution of Example 7 were tested by adding the test solution to a glass dish and lowering the disposable capillary tip and temperature probe into the solution. The dynamic setting was selected, and the instrument was set to collect 30 data points, the bubble age increasing from about 10 ms to about 30,000 ms.

[0105] Bubble tensiometry was used to calculate the Critical Micelle Concentration (CMC) value for each of the solutions. The results are set forth below in Table 4.

Table 4

[0106] Each of solutions RM12A, RM13A, RM14A-RM14C, and RM15A were tested using the above procedures and are presented in Figures 1-7.

[0107] Figure 1 presents a comparison of the bubble age to surface tension for RM14A at CMC, sub-CMC, and super-CMC concentrations. Figure 2 presents a comparison of the bubble age to surface tension for RM14B at CMC, sub-CMC, and super-CMC concentrations. Figure 3 presents a comparison of the bubble age to surface tension for RM14C at CMC, sub-CMC, and super-CMC concentrations. Figure 4 presents a comparison of the bubble age to surface tension for RM12A, RM13A, RM14A, and RM15A at super-CMC concentrations. Figure 5 presents a comparison of the bubble age to surface tension for RM12A, RM 13 A, RM 14 A, and RM15A at CMC concentrations. Figure 6 presents a comparison of the bubble age to surface tension for RM12A, RM13A, RM14A, and RM15A at CMC concentrations. Figure 7 presents a comparison of bubble age to surface tension for RM12A, RM 13 A, RM14A, and RM15A at sub-CMC concentrations.

[0108] It is notable that chlorhexidine di-[373-glucoheptonate and chlorhexidine di-[381- glucoheptonate salts appeared to cause surface tension to fall more rapidly and to a lower value than either the di-a-glucoheptonate or the comparative digluconate salt. This was an unexpected finding, and appears to indicate that the chlorhexidine di-[3-glucoheptonate salts associate in such a manner that allows for a greater reduction in surface tension. Without being bound to the theory, it is believed that there is an improvement in molecular-level kinetics or the electrostatic process, which allows for a faster reduction in the surface tension and generally a lower surface tension value.

[0109] The data further indicate that chlorhexidine di-[373-glucoheptonate and di-[381- glucoheptonate salts behave similarly. To achieve the desired results, it is believed that a [3- isomer concentration of at least about 25% should be used. In one embodiment, the composition comprises 25% [3-isomer, 75% a-isomer.

[0110] As demonstrated by the results discussed herein, it also appears that antimicrobial behavior favors chlorhexidine diglucoheptonate salts prepared with [3-isomers, and that biocidal activity exceeds the performance of the comparative standard, digluconate salts. Therefore, without being bound by the theory it is believed that dynamic surface tension may be a useful tool for pre-screening of compounds that are capable of reducing surface tension, at either sub-CMC, CMC, or super-CMC concentrations, and ultimately, in determining the usefulness of a compound as a disinfectant.

Example 9: Conductivity Testing

[oni] Conductivities were measured using a VWR 23609-216 Pure H2O Tester conductivity meter. The meter probe was submerged into the test solution and swirled gently. Once the meter settled, the value was recorded. [0112] The conductivity of the solutions of Example 7 were tested at dilutions ranging from 0.1 M to 0.001 M. The data was plotted on a logarithmic scale and analyzed in the style of Heard and Ashworth . For each sample, three graphs were reported and fitted with a trend curve : outer data points, inner data points, all data points. This data was used as set forth below to calculate the Critical Micelle Concentration (CMC). An average CMC value was determined from the three trend samples (outer data points, inner data points, all data points) in order to more accuracy predict the CMC value.

[0113] Figures 8-10 reports the conductivity for RM14A as the outer data points, inner data points, and all data points, respectively.

[0114] LINEST was utilized to determine the slope and intercept for each line, and a combination of INTERCEPT and SLOPE was used to determine the intersection of the two fitted lines. The Critical Micelle Concentration (CMC) was determined for each of these plots by the intercept of the two lines. Set forth below in Table 5 is the calculated CMC for each graph and the average CMC.

Table 5

Example 10: Refractive Index Analysis

[0115] Refractive indices were measured using a Mettler Toledo RM40 LiquiPhysics Excellence temperature-controlled refractometer. The prism was covered by several drops of test solution and the cover lowered. The refractive indices were collected at 25 °C.

[0116] Decreasing molar concentrations of each sample (i.e. 0.10 M, 0.08 M, 0.06 M, etc.) were tested to determine the refractive index of the respective sample. RM12A, RM12B, and RM12C are reported in Figure 11. RM13A, RM13B, and RM13C are reported in Figure 12. RM14A (reported as 2.0: 1), RM14B (reported as 2.3: 1), and RM14C (reported as 2.6: 1) are shown in Figure 13. RM15A is reported in Figure 14.

Example 11 : NMR Analysis

[0117] In a further experiment, the concentration of a-glucoheptonic acid and [373- glucoheptonic acid in the aqueous solutions of Example 2 and 4, respectively, was evaluated by NMR analysis. Each respective aqueous solution was combined with an approximately equal mass of deuterium oxide. An excess of sodium hydroxide (45% aqueous solution) was added and then approximately 0.17 equivalents of sodium acetate was added to the solution to form a mixture. The mixture was stirred until completely homogeneous, and subjected to NMR analysis. NMR data was collected using a BRUKER Avance-III 400 MHz.

[0118] Characteristic peaks in the 'H-NMR spectra for glucoheptonate and acetate were identified and integrated. The integrations were converted to moles using the nominal integral for each peak, and then compared in order to calculate the molar ratio of glucoheptonate to acetate. This ratio was then used to calculate the starting concentration of glucoheptonic acid in solution. A comparison of weight percentage of solids (measured at 65°C) and % activity (via 'H-NMR) for select samples is set forth below in Table 6.

Table 6

Example 12: NMR Analysis

[0119] In a further experiment, RM13A and RM14A of Example 7 were subjected to NMR analysis. RM13A or RM14A were pipetted onto separate glass dishes and allowed to dry overnight in a 65°C forced air oven. A small magnetic stirbar and deuterium oxide were then added to each dish. The mixture was stirred until completely homogeneous, and subjected to NMR analysis. NMR data was collected using a BRUKER Avance-II 300 MHz.

[0120] NMR analysis was also used to determine the glucoheptonate to chlorhexidine molar ratio in RM13A and RM14A. Aqueous samples of RM13A and RM14A were diluted to approximately 10% in deuterium oxide and subjected to NMR analysis. NMR data was collected using a BRUKER Avance-II 300 MHz. Characteristic peaks in the 'H-NMR spectra for chlorhexidine and glucoheptonate were identified and integrated. The integrations were converted to moles using the nominal integral for each peak, and then compared in order to calculate the molar ratio of glucoheptonate to chlorhexidine.

Example 13: Antimicrobial Testing - Chlorhexidine Salts

[0121] Select compounds were tested for antimicrobial activity using ASTM method E2315- 16 (Standard Guide for Assessment of Antimicrobial Activity using a Time-Kill Procedure). Six microbes were tested: Candida albicans, Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Salmonella abony, and Listeria monocytogenes .

[0122] Four chlorhexidine salts were tested: chlorhexidine di-a-glucoheptonate (Sample G), chlorhexidine di-|373 -glucoheptonate (Sample H), chlorhexidine digluconate (Sample I), and chlorhexidine diacetate (Sample P). Both chlorhexidine diglucoheptonate salts were prepared at a 2: 1 molar ratio of glucoheptonic acid to chlorhexidine. Dilutions of each salt were prepared using sterile water (20-30 ppm hardness), to achieve the desired wt.% concentration.

[0123] Table 7 sets forth the Log Reduction data at 30 seconds for varying concentrations of each sample.

Table 7

[0124] As illustrated in this table, numerous tests exceeded the >99.9% kill standard at 30 seconds by exhibiting a Log Reduction of at least about 3. EPA methodologies generally consider a product to be a sanitizer when there is a Log 3 Reduction in the microbial population at five minutes, using ASTM test method El 153. [0125] Both glucoheptonate salts demonstrated efficacy in the ASTM E2315-16 Time-Kill Test. However, at lower concentrations, the di-[373-glucoheptonate salt demonstrated better performance than the di-a-glucoheptonate salt.

[0126] The di-a-glucoheptonate salt performed very strongly against Candida albicans and Listeria monocytogenes, and, like the di-[373-glucoheptonate salt, outperformed both chlorhexidine digluconate and diacetate. The dramatic increase in performance may be related to aggregation phenomena that are known to take place with chlorhexidine salts at increasing concentrations.

[0127] Samples G, H, I, and P were also tested at a concentration of 0.60 wt% active and evaluated for the average Log Reduction in microbial population at 10 minutes, across all microbes. The results are set forth below in Table 8.

Table 8

[0128] A further test was conducted at a concentration of 1.0 wt% active and evaluated for the average Log Reduction in microbial population at 10 minutes, across all microbes. The results are set forth below in Table 9.

Table 9

Example 14: Antimicrobial Testing - Glucoheptonic Acids

[0129] Two glucoheptonic acids were evaluated for antimicrobial activity against the same six microbes as Example 13. Sample L was a-glucoheptonic acid. Sample M was [373- glucoheptonic acid. The results are set forth below.

Table 10: Sample L (a-glucoheptonic acid)

Table 11: Sample M ([373-glucoheptonic acid)

Example 15: Antimicrobial Testing - Benzalkonium Salts

[0130] Benzalkonium salts were also tested using the same procedure as set forth in Example 13. The samples were benzalkonium chloride (C12-C16) (Sample C), benzalkonium chloride (C12-C14) (Sample D), benzalkonium chloride in combination with a stoichiometric amount of a-glucoheptonate (Sample F), and benzalkonium chloride in combination with a molar excess of a-glucoheptonate (Sample N). Sample F was prepared by mixing and extensively heating a stoichiometric quantity (1: 1) of sodium a-glucoheptonate with benzalkonium chloride, typified by the C12-14 alkyl chain. Sample N was prepared by simply blending benzalkonium chloride, typified by the C12-16 chain, with a molar excess of sodium a-glucoheptonate (1.6: 1). Both Samples F and N contain sodium chloride. The results are set forth below in Table 12.

Table 12

[0131] The superior performance of Samples F and N at thirty seconds, are notable when comparing the number of tests in which the Log Reduction was at least 4 (i.e. removal of >99.99% of the microbial population). Samples F and N exceeded this threshold 19 times, whereas the comparative benzalkonium chlorides (Samples C and D) only exceeded the Log 4 threshold seven times.

[0132] A further antimicrobial test was conducted to determine the log reduction at 10 minutes. Samples C, D, F, and N are as defined above. The results are set forth in Table 13.

Table 13

[0133] Samples F and N again showed an improved result in Log Reduction as compared to the comparative benzalkonium chlorides (Samples C and D). [0134] When introducing elements of the present invention or the preferred embodiments(s) thereof, the articles "a", "an", "the" and "said" are intended to mean that there are one or more of the elements. The terms "comprising", "including" and "having" are intended to be inclusive and mean that there may be additional elements other than the listed elements.

[0135] In view of the above, it will be seen that the several objects of the invention are achieved and other advantageous results attained.

[0136] As various changes could be made in the above products and methods without departing from the scope of the invention, it is intended that all matter contained in the above description and shown in the accompanying drawings shall be interpreted as illustrative and not in a limiting sense.

Claims

- 32 -CLAIMS

1. A method of disinfecting or sanitizing, wherein the method comprises: contacting an aqueous composition comprising a component selected from the group consisting of glucoheptonic acid, chlorhexidine diglucoheptonate, benzalkonium glucoheptonate, and combinations thereof with a microbial population.

2. The method of claim 1, wherein the Log Reduction is about 3 or greater, 30 seconds after application.

3. The method of claim 1, wherein the microbial population is contacted with an aqueous composition comprising glucoheptonic acid.

4. The method of claim 3, wherein the microbial population is contacted with an aqueous composition comprising a-glucoheptonic acid and P-glucoheptonic acid and the composition comprises an a-glucoheptonic acid concentration, based on total isomers of glucoheptonic acid, of about 90% or greater.

5. The method of claim 1, wherein the microbial population is contacted with an aqueous composition comprising chlorhexidine di-a-glucoheptonate.

6. The method of claim 1, wherein the microbial population is contacted with an aqueous composition comprising chlorhexidine di-|373 -glucoheptonate.

7. The method of claim 1, wherein the microbial population is contacted with an aqueous composition comprising chlorhexidine di-|381 -glucoheptonate.

8. The method of claim 1, wherein the microbial population is contacted with an aqueous composition comprising chlorhexidine di-a-glucoheptonate and chlorhexidine di-P- glucoheptonate and the composition comprises a chlorhexidine di-a-glucoheptonate concentration, based on total isomers of chlorhexidine diglucoheptonate, of about 90% or greater.

9. The method of claim 1, wherein the microbial population is contacted with an aqueous composition comprising chlorhexidine di-a-glucoheptonate and chlorhexidine di~P~ glucoheptonate and the composition comprises a chlorhexidine di-P-glucoheptonate concentration, based on total isomers of chlorhexidine diglucoheptonate, of about 25% or greater. - 33 -

10. The method of claim 1, wherein the microbial population is contacted with an aqueous composition comprising benzalkonium glucoheptonate.

11. The method of any one of claims 1 to 10, wherein the microbial population comprises C. albicans; S. aureus, S. epidermidis, E. coli, S. abony, L. monocytogenes, or combinations thereof.

12. The method of any one of claims 1 to 10, wherein the microbial population comprises SARS-CoV-2.

13. A method of preparing glucoheptonic acid, the method comprising: dissolving a glucoheptonate salt in water to form a solution; combining the solution with an acidic ion exchange resin to form a slurry; filtering the slurry to remove the ion exchange resin to form glucoheptonic acid; wherein the conversion of the glucoheptonate salt to glucoheptonic acid is about 99% or greater, about 99.1% or greater, about 99.2% or greater, about 99.3% or greater, about 99.4% or greater, or about 99.5% or greater.

14. The method of claim 13, wherein the slurry is dried to form the glucoheptonic acid.

15. The method of claim 13, wherein the acidic ion exchange resin comprises a DOWEX MARATHON MSC and/or LEWATIT MONOPLUS SP 112 H ion exchange resin.

16. A method of preparing chlorhexidine diglucoheptonate, the method comprising: combining chlorhexidine and water to form a slurry; and adding the glucoheptonic acid of claim 13 to the slurry and mixing the combination until the glucoheptonic acid is dissolved.

17. The method of claim 16, wherein the chlorhexidine diglucoheptonate comprises a chlorhexidine di-a-glucoheptonate concentration, based on total isomers of chlorhexidine diglucoheptonate, of about 90% or greater.

18. The method of claim 16, wherein the chlorhexidine diglucoheptonate comprises a chlorhexidine di-P-glucoheptonate concentration, based on total isomers of chlorhexidine diglucoheptonate, of about 25% or greater.

19. The method of claim 16, wherein the chlorhexidine diglucoheptonate has a molar ratio of glucoheptonic acid to chlorhexidine of from about 1.5: 1 to about 3: 1, from about 2: 1 to about 2.9: 1, from about 2: 1 to about 2.8: 1, from about 2: 1 to about 2.7: 1, from about 2: 1 to about 2.6: 1, or from about 2.1 : 1 to about 2.6: 1.

20. The method of claim 10, wherein the benzalkonium glucoheptonate is prepared by a method comprising: combining a benzalkonium salt and a glucoheptonate salt in a container and stirring the combination.

21. The method of claim 20, wherein the benzalkonium salt is benzalkonium chloride and the glucoheptonate salt is sodium glucoheptonate.

22. The method of claim 20 or 21, wherein the benzalkonium salt and glucoheptonate salt are combined in the presence of a solvent.

23. A method of evaluating a compound for performance in microbial disinfecting or sanitizing, the method comprising: conducting a Bubble Pressure Tensiometry test of a compound; evaluating the reduction in surface tension exhibited by a compound as a function of time; and comparing the surface tension reduction to a compound known to exhibit microbial disinfecting or sanitizing properties to determine if the tested compound is useful for microbial disinfecting or sanitizing.

24. A method of disinfecting or sanitizing, wherein the method comprises: contacting an aqueous composition comprising a glucoheptonate salt with a microbial population; wherein the glucoheptonate salt is formed by process comprising reacting an intermediate glucoheptonate salt with a compound selected from the group consisting of benzethonium chloride, polymeric quaternary ammonium salts, dialkyldimethyl quaternary ammonium salts, dialkylmethyl ammonium quaternary ammonium salts with twin tails, other alkyl or hydroxyalkyl substituted quaternary ammonium salts, cetylpyridinium chloride, benzyl substituted quaternary ammonium salts, picolinium salts, imidazolinium salts, N-ethyl- Morpholinium salts, isoquinolinium salts, chlorhexidine salts, and combinations thereof.

25. A method of disinfecting or sanitizing, wherein the method comprises: contacting an aqueous composition comprising a glucoheptonate compound with a microbial population; wherein the glucoheptonate compound is prepared by combining a source of a glucoheptonate anion and a source of a cationic counterion selected from the group consisting of benzethonium chloride, polymeric quaternary ammonium salts, dialkyldimethyl quaternary ammonium salts, dialkylmethyl ammonium quaternary ammonium salts with twin tails, other alkyl or hydroxyalkyl substituted quaternary ammonium salts, cetylpyridinium chloride, benzyl substituted quaternary ammonium salts, picolinium salts, imidazolinium salts, N- ethyl-Morpholinium salts, isoquinolinium salts, chlorhexidine salts, and combinations thereof.

26. A method of disinfecting or sanitizing, wherein the method comprises: contacting an aqueous composition comprising a glucoheptonate compound with a microbial population; wherein the glucoheptonate compound is selected from the group consisting of benzethonium glucoheptonate, polymeric quaternary ammonium glucoheptonates, and cetylpyridinium glucoheptonate.

27. A method of disinfecting or sanitizing, wherein the method comprises: contacting an aqueous composition comprising a glucoheptonate compound with a microbial population; wherein the glucoheptonate compound is a quaternary ammonium salt having the formula: RIR2R3R4N⁺X", wherein Ri and R2 are independently selected from the group consisting of straight, branched, or cyclic alkyl or alkenyl groups having from about 8 to about 18 carbon atoms,

R3 and R4 are independently selected from the group consisting of straight alkyl groups having from about 1 to about 4 carbon atoms, hydroxyethyl, hydroxypropyl, hydroxybutyl, -(CH2CH2O)_mCH2CH2OH, -(CH2CHCH3O)_mCH₂CHCH3OH, - (CH2CH₂O)_m(CH2CHCH3O)_nCH2CHCH3OH, or - (CH2CHCH3O)m(CH2CH2O)nCH2CH₂OH-, wherein m is an integer from about 1 to about 10 and n is an integer from about 1 to about 10, and

X" is glucoheptonate. - 36 -

28. A method of disinfecting or sanitizing, wherein the method comprises: contacting an aqueous composition comprising a glucoheptonate compound with a microbial population; wherein the glucoheptonate compound is a quaternary ammonium salt having the formula: RIR2R3RIN⁺X‘, wherein Ri is selected from the group consisting of 3 -alkoxy-2 - hydroxypropyl or a straight, branched, or cyclic alkyl or alkenyl group having from about 8 to about 18 carbon atoms,

R2, R3 and R4 are independently selected from the group consisting of straight alkyl groups having from about 1 to about 4 carbon atoms, hydroxyethyl, hydroxypropyl, hydroxybutyl, -(CH2CH2O)_mCH2CH2OH, -(CH2CHCH3O)_mCH₂CHCH3OH, - (CH2CH₂O)_m(CH2CHCH3O)_nCH2CHCH3OH, or - (CH2CHCH₃O)m(CH2CH2O)_nCH2CH2OH, wherein m is an integer from about 1 to about 10 and n is an integer from about 1 to about 10, and

X" is glucoheptonate.

29. A method of disinfecting or sanitizing, wherein the method comprises: contacting an aqueous composition comprising a glucoheptonate compound with a microbial population; wherein the glucoheptonate compound is a quaternary ammonium salt having the formula: R^R^R-iN 'X-. wherein Ri is selected from the group consisting of a straight, branched, or cyclic alkyl or alkenyl group having from about 8 to about 18 carbon atoms,

R2 is selected from the group consisting of a substituted or unsubstituted benzyl, ethylbenzyl, naphthyl, or methylnaphthyl,

R3 and R4 are independently selected from the group consisting of straight alkyl groups having from about 1 to about 4 carbon atoms, hydroxyethyl, hydroxypropyl, hydroxybutyl, -(CH2CH2O)_mCH2CH2OH, -(CH2CHCH3O)mCH₂CHCH3OH, - (CH2CH₂O)_m(CH2CHCH3O)_nCH2CHCH3OH, or - (CH2CHCH3O)m(CH2CH2O)nCH2CH₂OH, wherein m is an integer from about 1 to about 10 and n is an integer from about Ito about 10, and - 37 -

X" is glucoheptonate.

30. A method of disinfecting or sanitizing, wherein the method comprises: contacting an aqueous composition comprising a glucoheptonate compound with a microbial population; wherein the glucoheptonate compound is a quaternary ammonium salt having the formula RIR2R RIN⁺X‘, wherein Ri is selected from the group consisting of 3 -alkoxy-2 - hydroxypropyl or a straight, branched, or cyclic alkyl or alkenyl group having from about 8 to about 18 carbon atoms,

R2 is selected from the group consisting of a substituted or unsubstituted benzyl, ethylbenzyl, naphthyl, or methylnaphthyl, a straight, branched, or cyclic alkyl or alkenyl group having from about 8 to about 18 carbon atoms, a straight alkyl group having from about 1 to about 4 carbon atoms, hydroxyethyl, hydroxypropyl, hydroxybutyl, - (CH2CH2O)_mCH2CH₂OH, -(CH₂CHCH3O)_mCH2CHCH₃OH, - (CH2CH₂O)_m(CH2CHCH3O)_nCH2CHCH3OH, or - (CH2CHCH3O)m(CH2CH2O)nCH2CH₂OH, wherein m is an integer from about 1 to about 10, n is an integer from about 1 to about 10,

R and R4 are independently selected from the group consisting of a straight alkyl group having from about 1 to about 4 carbon atoms, hydroxyethyl, hydroxypropyl, hydroxybutyl, -(CH2CH2O)_mCH2CH2OH, -(CH2CHCH3O)_mCH₂CHCH3OH, - (CH2CH₂O)_m(CH2CHCH3O)_nCH2CHCH3OH, or - (CH2CHCH₃O)m(CH2CH2O)_nCH2CH2OH, where m is an integer from about 1 to about 10 and n is an integer from about 1 to about 10, and

X" is glucoheptonate.

31. A method of disinfecting or sanitizing, wherein the method comprises: contacting an aqueous composition comprising a glucoheptonate compound with a microbial population; wherein the glucoheptonate compound is selected from the group consisting of

is glucoheptonate.