NATURAL ANTIBACTERIAL AND ANTIVIRAL BIOLOGICAL COMPOSITIONS, COMPOUNDS, METHOD FOR OBTAINING SAME, AND USE THEREOF

Abstract

A natural antibacterial and antiviral biological composition is provided for use in hospitals and homes, in particular a composition that includes ricinoleic acid derived from the extraction of castor oil, diethanolamine and propolis, a method for obtaining the composition, and the use thereof. The invention can be used to control various pathogenic microorganisms, such as bacteria belonging to the Staphylococcus, Pseudomonas, Enterobacter and Bacillus genera; as well as hospital-environment fungi (filamentous and yeast-like) and phytopathogenic fungi (Colletrotichum fragariae). The product can be sprayed on biotic and abiotic surfaces, and non-woven fabric can subsequently be used to remove organic matter, including microorganisms. The product can also be used to disinfect medical, dental and hospital items by immersion followed by quick rinsing.

Claims

1. A natural antibacterial and antiviral biological composition comprising: ricinoleic acid selected from a group consisting of: fatty acids, diethanolamine selected from a group consisting of amine dialcohol, and propolis selected from a group consisting of prenylated compounds.

2. The composition according to claim 1, wherein the composition is obtained by reacting ricinoleic acid, 2.0 to 4.0% by weight, diethanolamine from 1.0 to 1.5% by weight and propolis extract, from 0.2 to 1.0% by weight.

3. The composition according to claim 1, wherein the propolis used to prepare the extract is of a BRPX type.

4. The composition according to claim 1, wherein the propolis extract comprises one or more prenylated compounds, such as pcoumaric acid and derivatives.

5. The composition according to claim 1, wherein the composition is presented in a form of gel, ointment, lotions and liquid solution.

6. A method comprising: (a) weighing 0.1 to 0.3 g of propolis in a previously weighed 100 mL beaker; (b) weighing 2.0 to 7.0 g of ricinolamide in a previously weighed 50 mL beaker; (c) heating the ricinolamine to 50° C.; (d) pouring the ricinolamide into the 100 mL beaker where the propolis was weighed; (e) mixing gently with an aid of a glass stick; (f) placing water at 40° C. in the 50 mL beaker until the 50 mL beaker is almost full; (g) after the ricinolamide is fully mixed with the propolis, adding the water contained in the 50 mL beaker little by little to the mixture, gently shaking; (h) transferring the mixture to a 100 mL flask; (i) adding water to the 50 mL beaker and pouring into the 100 mL flask with an aid of a Pasteur pipette; (j) hitting a meniscus with water; and (k) gently shaking and transferring to a labeled flask.

7. A compound according to claim 1, wherein the compound is ricinolamide p-coumarate and has a structural formula: ##STR00007##

8. The compound according to claim 1, wherein the compound is ricinolamide 3-prenyl-4-hydroxycinnamate and has a structural formula: ##STR00008##

9. The compound according to claim 1, wherein the compound is ricinolamide 3,5-diprenyl-4-hydroxycinnamate and has a structural formula: ##STR00009##

10. The compound according to claim 1, wherein the compound is ricinolamide 2,2-dimethyl-8-prenyl-2H-1-benzopyran-6-propenoate and has a structural formula, ##STR00010##

11. The compound according to claim 7, wherein the compound has the following physicochemical properties (a) concentration of ricinolamide 2.0 to 7.0%; (b) propolis concentration 0.1 to 3.0%; (c) electrical conductivity 926.00±56.72 μS/cm; (d) Color 8.539±0.192 mm Pfund; (e) Density 1.0100±0.0027 g/cm.sup.3; (f) Fluidity (viscosity)—Runoff time: 28 s Value (0.49*(t-35)) CsT: 3.43 CentiPoise: 3.44; (g) pH of 7.5 to 8.5, pH between 8 and 8.3.

12. The compound according to claim 7, wherein the compound for disinfecting Staphylococcus, Pseudomonas, Enterobacter and Bacillus microorganisms; hospital-environment fungi (filamentous and yeast), phytopathogenic fungi (Colletrotichum fragariae), C. albicans, poliovirus type 1 of poliomyelitis, influenza, HIV, H1N1, viral hepatitis, herpes virus types 1 and 2, dengue hemorrhagic virus and coronavirus of animal origin.

13. A method comprising: utilizing the composition of claim 1 for preparing a biocide to eliminate pathogenic microorganisms.

Description

BRIEF DESCRIPTION OF DRAWINGS

[0040] Some of the embodiments will be described in detail, with references to the following Figures, wherein like designations denote like members, wherein:

[0041] FIG. 1 shows a delimited area to be tested;

[0042] FIG. 2 shows microbial growth collected on a surface of a surgical lighting after application of a biocide of embodiments of the invention (left) and without application (right);

[0043] FIG. 3 shows an analysis of the sample by high performance liquid chromatography (HPLC);

[0044] FIG. 4 shows main bioactive compounds in contact with water that form micellar structures;

[0045] FIG. 5A shows a container of ricinolamide;

[0046] FIG. 5B shows a container of ricinolamide with a soft extrast of propolis without alcohol;

[0047] FIG. 5C shows a container of an aqueous solution of ricinolamide with propolis; and

[0048] FIG. 5D shows a container of alcoholic tincture in water.

DETAILED DESCRIPTION

[0049] Embodiments of the present invention provide a totally clear natural biocidal composition resulting from the association of ricinoleic acid from the extraction of castor oil, diethanolamine and propolis in order to enhance antibacterial and antiviral activity. The compound resulting from the association is completely clear, as the propolis is easily incorporated into the mixture of ricinoleic acid and diethanolamine, without forming micelles that could result in a cloudy solution. Due to the detergent properties of the biocide, the dissolution of the propolis occurs, forming a clear solution, and a greater amount of propolis can be used in the formulation, depending on the use.

[0050] The composition comprises 1.4 to 5.0%, more 2.0 to 4.0%, by weight of ricinoleic acid. Ricinoleic acid is added to diethanolamine. The reaction of ricinoleic acid (A) with diethanolamine (B), produces an amide (C), called ricinolamide. In this reaction, the formation of the amide and the loss of water occur.

[0051] Below is the structure of the mixture of ricinoleic acid and diethanolamine, as well as the product Ricinolamide formed:

##STR00001##

[0052] Chemical reaction of ricinoleic acid (A) with diethanolamine (B) forming the amide called ricinolamide (C).

[0053] The ricinoleic acid is selected from commercial batches. The compound comprises from 1.4 to 5.0%, more 2.0 to 4.0%, of ricinoleic acid.

[0054] Diethanolamine is selected from commercial batches. The compound comprises from 0.5 to 2.0%, more from 1.0 to 1.5%, of diethanolamine.

[0055] Another aspect of embodiments of the present invention relates to a method for obtaining an amide compound with antibacterial and antiviral effect, comprising the steps of: [0056] (a) 25% diethanolamine (A) and 75% ricinoleic acid (B) are added to a reactor with mechanical agitation; [0057] (b) The two components (A)+(B) are mixed for 2 hours, monitoring the temperature, as it is an exothermic reaction; [0058] (c) The crude product is diluted in warm water, from 40° C. to 60° C., until its complete dissolution. The reaction is carried out without a catalyst, not exceeding 60° C., in a controlled environment at 20° C.; [0059] (d) Final dilution is done in water, resulting in a 50% solution.

[0060] If we consider the equimolar reaction (1 mol of ricinoleic acid reacts with 1 mol of diethanolamine forming 1 mol of the amide and 1 mol of water) we have: 75% ricinoleic acid (RA) at 95% purity reacting with 25% diethanolamine (DIET). As the molecular weight of RA is 298.46 g/mol and of DIET 105.14 g/mol and correcting the mass of RA (discounting impurities) we have: 71.25 g of RA which is 0.24 mol and 25.0 g of DIET which is 0.24 mol. Therefore, the compounds react in equimolar amounts to form 0.24 mol of the amide (C) plus 0.24 mol of water. We have the results shown in Table 6.

TABLE-US-00006 TABLE 6 Stoichiometric calculation of the Ricinolamide formation reaction. Reagents Products Name Weight (g) Name Weight (g) Yield (%) RA 71.25 Amide 92.54 95.5 DIET 25.00 Water 4.32 Impurities 3.75 Impurities 3.14 Total 100.0 Total 100.00

[0061] Therefore, the yield of the reaction of ricinoleic acid with diethanolamine, forming the amide, is 95.5%.

[0062] Chemical Characteristics of Propolis:

[0063] The propolis used in embodiments of the present invention is called BRPX, rich in prenylated compounds, as described below:

##STR00002##

[0064] Main markers found in BRPX propolis, with DHCA (Artepillin-C) being the majority.

[0065] Below are the quality parameters referring to the sample used in the present study: total phenolic compounds: 9.0 to 10.0%, total flavonoids between 2.5 and 3.5%, insoluble residues between 35.0 and 45.0%, ethanol soluble solids between 45.0 and 50.0%, moisture (loss on desiccation) between 4.0 and 5.0% and wax content between 9.0 and 10.0%.

[0066] In FIG. 3, analysis of the same sample by high performance liquid chromatography (HPLC) is shown.

[0067] The quantitative analysis of these markers showed levels of: p-coumaric acid (1) between 1.5 and 2.0%; PHCA (2) between 0.3 to 0.5%; Artepillin-C (3) between 3.0 and 5.0; DPB (4) between 0.3 to 5.0%. The antioxidant activity, evaluated by the DPPH radical decolorization method (Veiga et al., 2017) was between 5.0 and 15.0 ug/mL.

[0068] Propolis has great difficulty in solubilizing in water, forming micelles and clouding the solution. Some companies have launched propolis-based products on the market, stating on the label that they are aqueous extracts.

[0069] As the main characteristic of propolis is that it is a resinous substance, whose bioactive compounds are practically insoluble in water, except for p-coumaric acid and caffeoylquinic acids, it is not to be expected that clear solutions will be present. Due to the nonpolar characteristic of the main bioactive compounds, these in contact with water form micellar structures, as shown in FIG. 4. One way to solubilize propolis to form a clear solution is to add a base, such as sodium hydroxide, to water.

[0070] The acids in propolis (major compounds) react with the base, forming salt and water, but the final solution, although clear, has basic characteristics, with a pH between 8.0 and 9.0, not being suitable for human consumption.

[0071] FIG. 4—The circled area corresponds to a “cream” formed on the walls of the container, indicating the incompatibility of propolis with water.

[0072] The propolis should be present at 0.1 to 3.0%, more from 0.2 to 1.0%, by weight of the composition.

[0073] The biocidal compound, object of embodiments of this invention, can be used to develop various sanitizing detergent products for domestic, agricultural (cleaning machinery, floors, surfaces and animals), hospital (sanitization and disinfection) use. Examples include the presentation of products in the form of lotions, gels, sprays, shampoos, etc.

[0074] Antibacterial and antiviral benefits are provided by the fast-acting compound of embodiments of the invention and therefore it is especially suitable for incorporation into biocidal products.

[0075] It is desired that the compound of embodiments of the invention be formulated to have a pH of 7.5 to 8.5, between 8 and 8.3, wherein the effectiveness of the synergistic interaction between ricinoleic acid, diethanolamine and propolis is seen as maximum.

[0076] The biocidal compound can also be used to clean and disinfect topical areas such as wounds and therefore can be formulated as a gel or ointment.

Formulations of the Invention

[0077] Control ricinolamide at 10% (aqueous solution): 10 g of the product in 100 mL (final volume in a volumetric flask, the meniscus is adjusted with water). Amount of the active: 9.55%.

[0078] Control propolis at 3% (solution in ethanol or DMSO): 3 g of the product in 100 mL (final volume in a volumetric flask, the meniscus is adjusted with ethanol or DMSO). Amount of the actives: p-coumaric acid (1) from 0.045 to 0.06% (450 to 600 ug/mL); 3-prenyl hydroxycinnamic acid (2) from 0.009 to 0.015% (90 to 150 μg/mL); 3,5-diprenyl hydroxycinnamic acid (3) from 0.09 to 0.15% (900 to 1500 μg/mL); 2,2-dimethyl-8-prenyl-2H benzopyran-6-propenoic acid (4) between 0.009 to 0.15% (90 to 1500 μg/mL).

[0079] Biocidal Composition 1: 0.2% propolis and 5% ricinolamide: 5 g of ricinolamide and 0.2 g of propolis. Amount of the active ricinolamide: 4.8%. Amount of actives: p-coumaric acid (1) from 0.003 to 0.004% (30 to 40 μg/mL); 3-prenyl-4-hydroxycinnamic acid (2) from 0.0006 to 0.001% (6 to 10 μg/mL); 3,5-diprenyl-4-hydroxycinnamic acid (3) between 0.006 to 0.01% (60 to 100 μg/mL); 2,2-dimethyl-8-prenyl-2H-1-benzopyran-6-propenoic acid (4) between 0.0006 to 0.01% (6 to 100 μg/mL).

[0080] Biocidal Composition 2: 0.5% of propolis and 3% of ricinolamide: 3 g of ricinolamide and 0.5 g of propolis. Amount of the active ricinolamide: 2.86%. Amount of actives: p-coumaric acid (1) from 0.0075 to 0.01% (75 to 100 μg/mL); 3-prenyl-4-hydroxycinnamic acid (2) from 0.0015 to 0.0025% (15 to 25 μg/mL); 3,5-diprenyl-4-hydroxycinnamic acid (3) between 0.015 to 0.025% (150 to 250 μg/mL); 2,2-dimethyl-8-prenyl-2H-1-benzopyran-6-propenoic acid (4) between 0.0015% to 0.025% (15 to 250 μg/mL).

[0081] Biocidal Composition 3: 0.1% propolis and 7% ricinolamide: 7 g of ricinolamide and 0.1 g of propolis. Amount of the active ricinolamide: 6.68%. Amount of actives: p-coumaric acid (1) from 0.0015 to 0.002% (15 to 20 μg/mL); 3-prenyl-4-hydroxycinnamic acid (2) from 0.0003 to 0.0005% (3 to 5 μg/mL); 3,5-diprenyl-4-hydroxycinnamic acid (3) between 0.003 to 0.005% (30 to 50 μg/mL); 2,2-dimethyl-8-prenyl-2H-1-benzopyran-6-propenoic acid (4) between 0.0003 to 0.005% (3 to 50 μg/mL).

[0082] Biocidal Composition 4: 3% propolis and 2% ricinolamide: 2 g of ricinolamide and 3 g of propolis. Amount of the active ricinolamide: 1.90%. Amount of actives: p-coumaric acid (1) from 0.045 to 0.06% (450 to 600 μg/mL); 3-prenyl-4-hydroxycinnamic acid (2) from 0.009 to 0.015% (90 to 150 μg/mL); 3,5-diprenyl-4-hydroxycinnamic acid (3) from 0.09 to 0.15% (900 to 1500 μg/mL); 2,2-dimethyl-8-prenyl-2H-1-benzopyran-6-propenoic acid (4) between 0.009 to 0.15% (90 to 1500 μg/mL).

[0083] Obtaining Method:

[0084] a) Weigh 0.1 to 3.0 g of propolis in a previously weighed 100 mL beaker;

[0085] b) Weigh 2.0 to 7.0 g of ricinolamide in a previously weighed 50 mL beaker;

[0086] c) Heat the ricinolamide to 50° C.;

[0087] d) Pour the ricinolamide into the 100 mL beaker where the propolis was weighed;

[0088] e) Mix gently with the aid of a glass stick;

[0089] f) Place water at 40° C. in the 50 mL beaker until it is almost full;

[0090] g) After the ricinolamide is fully mixed with the propolis, add the water contained in the 50 mL beaker little by little to this mixture, gently shaking;

[0091] h) Transfer the mixture to a 100 mL flask;

[0092] i) Add water to the 50 mL beaker and continue pouring into the 100 mL flask with the aid of a Pasteur pipette;

[0093] j) Hit the meniscus with water;

[0094] k) Shake gently and transfer to a labeled flask.

[0095] Molecular Structures of Compounds

[0096] Compound 1: C.sub.31H.sub.49NO.sub.6—Exact mass: 531.36 g/mol

##STR00003##

ricinolamide p-coumarate

[0097] Compound 2: C.sub.36H.sub.57NO.sub.6—Exact mass: 599.42 g/mol

##STR00004##

ricinolamide 3-prenyl-4-hydroxycinnamate

[0098] Compound 3: C.sub.41H.sub.65NO.sub.6—Exact mass: 667.48 g/mol

##STR00005##

ricinolamide 3,5-diprenyl-4-hydroxycinnamate

[0099] Compound 4: C.sub.4H.sub.65NO.sub.6—Exact mass: 667.48 g/mol

##STR00006##

Ricinolamide 2,2-dimethyl-8-prenyl-2H-1 -benzopy ran-6-propenoate

[0100] Ricinolamide (FIG. 5A) mixes easily with the dry ethanolic extract of propolis, called “soft extract”. It forms a solution with oily characteristics (FIG. 5B). The process of preparing the mixture must be done with very gentle agitation so as not to incorporate air bubbles.

[0101] FIG. 5—Containers showing the ricinolamide (FIG. 5A), the ricinolamide with the soft extract of propolis without alcohol (FIG. 5B), the aqueous solution of the ricinolamide with propolis (FIG. 5C) and the alcoholic tincture of propolis in water (FIG. 5D).

[0102] It was found, quite unexpectedly, that the mixture of ricinolamide with the soft extract of propolis without alcohol and then diluted in water, forms a clear solution, as seen in FIG. 5C unlike the alcoholic extract of propolis in contact with water, FIG. 5D

[0103] Obtaining this clear solution FIG. 5C is characterized by the physical-chemical parameters mentioned in Table 2.

[0104] Although the present invention has been disclosed in the form of embodiments and variations thereon, it will be understood that numerous additional modifications and variations could be made thereto without departing from the scope of the invention.

[0105] For the sake of clarity, it is to be understood that the use of ‘a’ or ‘an’ throughout this application does not exclude a plurality, and ‘comprising’ does not exclude other steps or elements.