WO2023170183A1 - Composition for treatment of parasitic or viral infection in alimentary and/or respiratory tract - Google Patents
Composition for treatment of parasitic or viral infection in alimentary and/or respiratory tract Download PDFInfo
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- WO2023170183A1 WO2023170183A1 PCT/EP2023/055944 EP2023055944W WO2023170183A1 WO 2023170183 A1 WO2023170183 A1 WO 2023170183A1 EP 2023055944 W EP2023055944 W EP 2023055944W WO 2023170183 A1 WO2023170183 A1 WO 2023170183A1
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Classifications
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- A61K36/18—Magnoliophyta (angiosperms)
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- A61K36/54—Lauraceae (Laurel family), e.g. cinnamon or sassafras
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Definitions
- the present invention relates to compositions comprising at least one Cl - C12 organic acid and Ceylon cinnamon and its use wherein the organic acid is a carboxylic acid with acidic properties.
- Organic acids are widely used for different purposes in different combinations all over the world. It’s used as a disinfectant, as a food/feed preservation technique, as a growth promoter in production animals and many more. Organic acids have been successfully used as a growth promoter in pigs for more than 25 years and they have also been found to be effective in poultry production. From the use of organic acids in poultry and pigs, one can expect an improvement in performance similar to or better than that of antibiotic growth promoters, without the public health concern.
- the present invention aims to solve one of the aforementioned problems and provides a composition comprising at least one Cl -Cl 2 organic acid and/or its salts in combination with an iron component , Ceylon cinnamon and B-vitamins and wherein the organic acid is a carboxylic acid with acidic properties.
- the present inventors have solved the above-mentioned need by providing in a first aspect a composition comprising at least one C1-C12 organic acid and/or its salt, an iron component, Ceylon cinnamon and B-vitamins in an amount from 0 to 200 mg/g dry weight of the composition and in amounts corresponding to at least that which theoretically can be consumed during metabolism of the COOH-groups present wherein the organic acid is a carboxylic acid with acidic properties according Bronsted definition of an acid which is a molecule that can donate a proton to another molecule.
- composition does not comprise L-carnitine.
- the organic acid is a C1-C8 and/or its salts wherein the total amount of the organic acids is in the range from 66 % (w/w) to 100% (w/w) by weight based on dry weight of the composition.
- the organic acid is a C1-C8 and/or its salts wherein the total amount of the organic acids is in the range from 66 % (w/w) to 99% (w/w) by weight based on dry weight of the composition, such as from 60% w/w to 95% w/w, such as from 75% w/w to 85% w/w.
- the organic acid is a fatty acid and/or its salts.
- the salt is selected from a Sodium salt or a Potassium salt.
- the organic acid is a Cl -Cl 2 fatty acid selected from short chain fatty acids, medium chain fatty acids and long chain fatty acids.
- the organic acid is a C1-C8 fatty acid.
- the fatty acids are selected from short chained fatty acids (SCFA): formic acid (Cl), acetic acid (C2), Propionic acid (C3), Butyric acid (C4), Fumaric acid (C4); Medium-chained fatty acids (MCFA): Sorbic acid (C6), Caprylic acid (C8) and Long-chained fatty acids (LCFA): Lauric acid (C12).
- SCFA short chained fatty acids
- Cl formic acid
- C2 acetic acid
- Propionic acid C3
- Butyric acid C4
- Fumaric acid C4
- MFA Medium-chained fatty acids
- MCFA Sorbic acid
- C8 Caprylic acid
- LCFA Long-chained fatty acids
- the at least one organic acid is formic acid and its salt.
- the iron component is ferrous fumarate.
- the fatty acids are selected from Caprylic acid (C8), Sorbic acid (C6), formic acid (Cl), Propionic acid (C3), Butyric acid (C4), acetic acid (C2) and any combinations thereof.
- the iron component is in an amount of 0 to 3.5% (w/w) by weight based on dry weight of the composition and the iron compound is preferably ferrous fumarate,
- the iron component is in an amount of 0.5 to 3.5% (w/w) by dry weight of the composition.
- the iron component is in an amount of 0.5 to 3.5% (w/w) by dry weight of the composition and the iron compound is ferrous fumarate.
- the composition of B-vitamins comprises 0-10 mg/g Bl, 0-25mg/g B2, 0-40 mg/g B3, 0-25 mg/g B5, 0-60 mg/g B6, 0-25 mg/g B7, 0-60 mg/g B9 and 0-250 pg/g B12 based on dry weight of the composition.
- the composition of B-vitamins comprises 2-10 mg/g Bl, 5-25mg/g B2, 10-25 mg/g B6, 0-25 mg/g B7, 0.1-60 mg/g B9 and 1-250 pg/g B12 based on dry weight of the composition.
- the composition of B-vitamins comprises 0.5-10 mg/g Bl, 0.5-25mg/g B2, B6 0.5-25 mg/g, 0-25 mg/g B7, 0.1-60 mg/g B9 and 10-250 pg/g B12 based on dry weight of the composition.
- the amount of vitamins, B6, B9 and B 12 is in the range from 10-50mg/g dry weight of the composition and the amount of B9 and B12 should at least correspond to that which can be consumed during metabolism of the COOH-group of the carboxylic acids in particular in combination with formic acid and is salt.
- the Ceylon cinnamon is bark from Cinnamomum verum and is in an amount from 0.1 to 6% (w/w) by dry weight of the composition, preferably in an amount from 0.5 to 6% (w/w) by dry weight of the composition.
- the composition comprises the composition further comprises a desiccant in an amount of 0-6% (w/w) by dry weight of the composition, wherein the desiccant is selected from Silicone dioxide, Magnesium oxide or any combinations thereof.
- the composition comprises an antioxidant in an amount of 0-1% (w/w) by dry weight of the composition, wherein the antioxidant is preferably selected from Vitamin C, Vitamin E or any combinations thereof.
- the amount of salt and carboxylic acids will give a pH amounting to 2.0 to 7.5 when the composition is in form of a suspension.
- the amount of salt and carboxylic acids will give a pH amounting to 2.0 to 7.5 when the composition is dissolved in water.
- the amount of salt and carboxylic acids will give a pH amounting to 2.0 to 6.0 when the composition is in form of a suspension.
- the amount of salt and carboxylic acids will give a pH amounting to 2.0 to 6.0 when the composition is dissolved in water.
- the composition is a dietary supplement.
- the composition is a pharmaceutical composition.
- the composition is administrated in the state of powder, suspension, gel-suspension, supplemented directly to food or feed or is swallowed as a tablet or capsule.
- the dietary supplement for decreasing infectivity of a virus and/or a parasite in a subject .
- dietary supplement for increasing virus and/or parasite resistance in a subject.
- a pharmaceutical composition for use in reducing the risk of and/or aid in treatment of a parasitic infection and/or viral infection in a subject.
- the parasite is caused by a parasite with a hard-shelled life cycle stage where it forms a cyst and/or oocyst.
- the parasite and/or the virus is comprised in the gastrointestinal system such as the alimentary tract and/or respiratory system of the subject.
- the virus is selected from rotaviruses, adenoviruses, coronaviruses, Hepatitis A, Hepatitis E, Norovirus, Enterovirus and Poliovirus.
- the parasite is selected from coccidiosis and giardiasis.
- the fatty acids are selected from Sodium Caprylate, Potassium Sorbate, Sodium Formate, Sodium Butyrate, Sodium Acetate, Sodium Propionate and any combinations thereof wherein the composition is for use in preventing or treatment of a parasite comprised in the gastrointestinal system such as the alimentary tract and/or respiratory system of the subject.
- the fatty acids are selected from Sodium Caprylate, Potassium Sorbate, Sodium Formate, Sodium Butyrate, Sodium Propionate and any combinations thereof wherein the composition is for use in preventing or treatment of a virus is comprised in the gastrointestinal system such as the alimentary tract and/or respiratory system of the subject.
- the subject is an animal, a fish or a human.
- the animal is selected from a ruminant, a horse, a race pigeon, a dog and a cat.
- the composition is administrated as a daily dose in the range 25-500 mg dry weight / kg body weight or 0.5-10g per 20kg body weight
- the first aspect is administrated as a daily dose of about 1g per 20kg body weight for use in reducing the risk of a parasitic infection and/or viral infection in a subject.
- the composition is administrated as a daily dose of 2g- 10g per 20kg body weight for use in treatment of or aid in treatment of a parasitic infection and/or viral infection in a subject.
- a method decreasing infectivity of a virus and/or a parasite in a subject comprising administering to the subject the composition according to the first aspect in a daily dose in the range from 25 to 500 mg dry weight / kg body weight of the subject.
- a method for use in reducing the risk of and/or aid in treatment of a parasitic infection and/or viral infection in a subject comprising administering to the subject the composition according to the first aspect in a daily dose in the range from 25 to 500 mg dry weight / kg body weight of the subject.
- a composition comprising one or more C1-C12 organic acid and/or its salt, an iron component, and B-vitamins for use in reducing the risk of and/or treatment of a parasitic infection and/or viral infection in a subject, wherein the B-vitamins is in an amount of 0-200 mg/g dry weight of the composition and the amount of B-vitamins corresponding to at least that which theoretically can be consumed during metabolism of the COOH-groups of the organic acid and wherein the organic acid is a carboxylic acid wherein the organic acid is a carboxylic acid with acidic properties according Bronsted definition of an acid which is a molecule that can donate a proton to another molecule.
- the composition does not comprise D/L-carnitine.
- D/L-carnitine is derived from lysine and methionine and is involved in metabolism of fatty acids.
- the Cl -Cl 2 organic acid is a fatty acid and/or its salts.
- the iron component is ferrous fumarate.
- the fatty acids are selected from Caprylic acid (C8), Sorbic acid (C6), formic acid (Cl), Propionic acid (C3), Butyric acid (C4), acetic acid (C2) and any combinations thereof.
- the at least one organic acid is formic acid and its salt.
- the fatty acids are selected from Sodium Caprylate, Potassium Sorbate, Sodium Formate, Sodium Butyrate, Sodium Propionate and any combinations thereof
- composition is administration by inhalation or intranasal application in the state of powder, suspension, gelsuspension, supplemented directly to food/feed or is swallowed as a tablet or capsule.
- the parasite is caused by a parasite with a hard-shelled life cycle stage where it forms a cyst and/or oocyst.
- the parasite and/or the virus is comprised in the gastrointestinal system such as the alimentary tract and/or respiratory system of the subject.
- the virus is selected from rotaviruses, adenoviruses, coronaviruses, Hepatitis A, Hepatitis E, Norovirus, Enterovirus and Poliovirus.
- the parasite is selected from coccidiosis and giardiasis.
- composition is an antiviral nasal spray.
- the present invention provides a composition comprising at least one Cl -Cl 2 organic acid and/or its salts in combination with an iron component, Ceylon cinnamon and B-vitamins and wherein the organic acid is a carboxylic acid with acidic properties.
- C1-C12 organic acid refers to a fatty acid compound comprising from 1 carbon atom to 12 carbon atoms and having acidic properties.
- Acidic property is defined herein as a molecule that can donate a proton to another molecule according to Bronsted definition of acids, Bronsted, J. N. (1923).
- Cl -Cl 2 carboxylic acids are selected from short chained fatty acids (SCFA): formic acid (Cl), acetic acid (C2), Propionic acid (C3), Butyric acid (C4), Fumaric acid (C4); Medium-chained fatty acids (MCFA): Sorbic acid (C6), Caprylic acid (C8) and Long-chained fatty acids (LCFA): Lauric acid (C12).
- SCFA short chained fatty acids
- Cl formic acid
- C2 acetic acid
- Propionic acid C3
- Butyric acid C4
- Fumaric acid C4
- MCFA Medium-chained fatty acids
- MCFA Sorbic acid
- Caprylic acid C8
- LCFA Long-chained fatty acids
- At least one of the carboxylic acids is formic acid and its salt.
- the fatty acids are selected from Caprylic acid (C8), Sorbic acid (C6), formic acid (Cl), Propionic acid (C3), Butyric acid (C4), acetic acid (C2) and any combinations thereof.
- the present composition comprises Cinnamon, Sodium Caprylate, Potassium Sorbate, Sodium Formate, Sodium Butyrate, Sodium Propionate and any combinations thereof in a dietary supplement for increasing virus resistance in a subject wherein the virus is comprised in the gastrointestinal system such as the alimentary tract and/or respiratory system and wherein the virus is selected from rotaviruses, adenoviruses, coronaviruses, Hepatitis A, Hepatitis E, Norovirus, Enterovirus and Poliovirus.
- the composition comprises Cinnamon, Sodium Caprylate, Potassium Sorbate, Sodium Formate, Sodium Butyrate, Sodium Propionate and any combinations thereof and in the form of a dietary supplement for decreasing infectivity of a virus and/or a parasite in a subject virus is comprised in the gastrointestinal system such as the alimentary tract and/or respiratory system and wherein the virus is selected from rotaviruses, adenoviruses, coronaviruses, Hepatitis A, Hepatitis E, Norovirus, Enterovirus and Poliovirus.
- the composition comprises cinnamon, Sodium Caprylate, Potassium Sorbate, Iron Fumarate, Sodium Formate, Sodium Butyrate, Sodium Acetate, Sodium Propionate and any combinations thereof
- Ceylon cinnamon as applied herein is bark from the Cinnamomum verum tree. Ceylon cinnamon from the Cinnamomum verum tree is often called true cinnamon and comprises lower amount of the toxin coumarin compared to Cassia cinnamon. The most produced cinnamon oil comes from Cassia cinnamon. Ceylon cinnamon is used herein.
- Cinnamon oil can be made from either the bark or the leaves which will also give different ratios between the different essential oils. The composition of these two is very different in relation to the relationship between the different essential oils.
- Cisa cinnamon contains approx. 250x more coumarin which in excessive doses can be toxic to the liver and kidneys. Too high doses of coumarin can also prevent Vitamin K synthesis in the gut and "Vitamin K bleeding deficiency" The toxic effects of Coumarin probably affect most people.
- Cinnamon oil from leaves often contains a lot of eugenol and very little cinnamaldehyde, while it is the other way around when using bark.
- Cinnamaldehyde has health-promoting effects by reducing the activity of important enzymes that trigger inflammatory reactions and allergic reactions.
- EP 1 691 626 Bl discloses recommended amounts of B6, B12, B9 and Fe for human and animals to be included in the composition of the present invention.
- viruses that are highly dependent on the fecal- oral route include Hepatitis A, Hepatitis E, Norovirus, Corona viruses (especially feline and equine coronaviruses), Enterovirus, Poliovirus and Rotavirus.
- Decrease infectivity is the reduction of a virus or parasites ability to infect a cell in a subject, i.e. increasing virus or parasitic resistance.
- Some of the most common causes for protozoan parasitic diarrhoea in dogs and cats include coccidiosis and giardiasis.
- Isospora and Cryptosporidium species are two of the most common causes of clinical manifestation of coccidiosis in dogs and cats.
- Other species may include Besnoitia, Toxoplasma, Hammondia and Sarcocystis.
- Coccidian parasites are known for their infectious oocysts with an obligate fecal-oral route of infection.
- Giardia parasites are another protozoan parasite with similar presentation of symptoms, however their life cycle do not include an oocyst.
- Wild and farmed fish are also sometimes infected with viruses and parasites that has a hard-shelled life cycle stage. Examples include many coccidian parasites like Calyptospora, Goussia, Eimeria, Epieimeria, Cryptosporidium, Crystallospora, and Octosporella(fungi).
- coccidian parasites like Calyptospora, Goussia, Eimeria, Epieimeria, Cryptosporidium, Crystallospora, and Octosporella(fungi).
- coccidiosis in farmed fish in Norway seems to be a problem rather in lumpfish than in other farmed fish like atlantic salmon and rainbow trout.
- Atlantic salmon and rainbow trout (salmonids)
- several viral diseases poses a threat to the industry.
- Cardiomyopathy syndrome Pancreas disease, Infectious salmon anemia, infectious pancreatic necrosis, Heart and skeletal muscle inflammation, Viral hemorrhagic septicaemia, Infectious hematopoietic necrosis and Salmon gill pox virus.
- protozoan diseases There are many different antiprotozoal agents commercially available. This is a class of pharmaceuticals used in the treatment of protozoan diseases. Some of the most important protozoan diseases include amebiasis, giardiasis, cryptosporidiosis, microsporidiosis, malaria, babesiosis, trypanosomiasis, Chagas disease, leishmaniasis and toxoplasmosis. Currently, many of the treatments for these infections are limited by their toxicity and present invention may reducing the risk of and/or aid in the treatment of these.
- composition comprising as the basic component at least one C1-C12 organic acid and/or its salt, an iron component, Ceylon cinnamon and B-vitamins in 0-200mg/g dry matter and in amounts corresponding to at least that which theoretically can be consumed during metabolism of the COOH- groups present.
- composition also contains a desiccant and an antioxidant.
- composition may also be administered in a suspension with pH amounting to 2-7.5.
- the composition contains 66-100% of one or more C1-C8 organic acids and/or its salts.
- the composition is characterized by an iron component consisting of 0-3.5% (w/w) iron, and the component preferably used is ferrous fumarate.
- composition may also contain B-vitamins in amounts of 0- 200mg/g dry weight of the composition, wherein 0-10 mg/g Bl, 0-25mg/g B2, 0- 40mg/g B3, 0-25mg/g B5, 0-60mg/g B6, 0-25mg/g B7, 0-60mg/g B9 and 0-250pg/g B12.
- composition is characterized by 0-6% cinnamon, preferably from the plant Cinnamomum vera.
- composition is characterized by 0-6% w/w desiccant, preferably Silicone dioxide and/or Magnesium oxide.
- composition is characterized by 0-1% w/w antioxidant, preferably Vitamin C or Vitamin E.
- composition is used to treat or to aid in treatment against infection by parasites that contain hard-shelled life cycle stages like cysts and oocysts.
- composition is used to treat or aid in treatment against infection by viruses with an affinity towards gastrointestinal system and respiratory system like rotaviruses, adenoviruses, coronaviruses, etc.
- composition is administrated by a daily dosage which may be applied, is in the range 25-500mg/kg bodyweight or 0.5-10g per 20kg body weight.
- composition is administration by inhalation or intranasal application in the state of powder, suspension, gel-suspension, supplemented directly to food/feed or is swallowed as a tablet or capsule.
- the main feature of the composition according to the invention comprises a basic component having at least one C1-C8 organic acid and/or its salt comprising at least formic acid, an iron component preferably iron fumarate, Ceylon cinnamon, and B6, B9 and B12-vitamins in the range from 0 to 200mg/g dry weight of the composition and in an amount corresponding to at least that which theoretically can be consumed during metabolism of the COOH-groups present and wherein the organic acid is a carboxylic acid having acidic properties and wherein the carboxylic acids are fatty acids and wherein the total amount of organic acids is at least 66% (w/w) by weight based on dry weight of the composition.
- the composition in addition may comprise antioxidants and desiccants and further components depending on its intended use wherein these further components don’t change the effect of the composition.
- Mortality in fish may be caused by several factors including viruses and parasites as discussed above.
- a study conducted on 600 Atlantic salmon showed a 100% reduction in mortality rate.
- the study design was constructed as follows:
- Feed A Control carrier pellet with no additives
- Feed B Carrier pellet with 1.5% of present invention comprising Sodium diformate, Calcium butyrate and Sodium caprylate.
- the fish were 40 grams and adapted to sea water at starting point.
- Results showed a 5% mortality rate in the control group after 9 and 10 weeks compared to zero mortality in group with carrier pellet. Growth and length were significantly improved within the first 5 weeks along with hepatic weight and intestinal tissue damage significantly reduced as well.
- Non-responders to placebo were crossed to present invention.
- the present invention contains salts of organic acids with established antibacterial effects, however results may suggest an even wider efficiency across viruses and parasites. Especially those involved in gastrointestinal illness.
- Oocysts were exposed to hypochlorite for 10 min on ice to remove bacterial contaminants and then washed twice in PBS before being resuspended in RPMI (a growth medium used in cell culture) at either pH 7 or at pH 3, the latter with or without (controls) different components from present invention. Single compounds were tested at concentrations of 5 mg/mL, and combinations with sodium formate at 2.5 mg/mL for each compound.
- the experimental set-up was as shown in Figure 1 (below).
- the study was run in triplicate (three individual set-ups), with each set-up consisting of technical triplicates of each experimental procedure.
- the copy number results obtained are shown in Table 1, and indicate the mean copy numbers of the qPCR target gene of C. parvum from the three replicate experiments. Higher numbers thus indicate greater numbers of parasites following infection and replication. Lower numbers indicate either reduced infection and/or reduced replication.
- Table 1 Percentages of inhibition of Cryptosporidium parvum infection and/or replication following incubation with different components, as related to incubation in RPMI pH 3 in the absence of these compounds.
- a laboratory study was conducted to test the effect of powder from cinnamon bark from Ceylon cinnamon on the infectivity of a virus (Experiment A and B) and to test the effect on the infectivity of a virus, using a combination of the following organic acids: Sodium format, Calcium format, iron fumarate and potassium sorbate (experiment C and D).
- the virus used for the experiments was Bovine Coronavirus (BCoV).
- BCoV Bovine Coronavirus
- the infectivity of the virus was tested with the use of human colorectal adenocarcinoma cells (HCT-8) and the infectivity of the virus was evaluated using microscope.
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Abstract
The present invention relates to a combination of different components and its use, where the invention comprises as the basic component at least one organic acid and/or its salt, an iron component, Ceylon cinnamon and B-vitamins. The combination may also contain a desiccant and an antioxidant. The invention will have a pH in the range of 2.0-7.5 when dissolved into a solution and is intended for all animals including primates. The present invention can be used in animal feed production, but can also be administered through other applications or as is. The daily dosage which may be applied, is in the range 25-500mg/kg body weight or0.5- 10g per 20kg body weight.
Description
COMPOSITION FOR TREATMENT OF PARASITIC OR VIRAL INFECTION IN
ALIMENTARY AND/OR RESPIRATORY TRACT
FIELD OF THE INVENTION
The present invention relates to compositions comprising at least one Cl - C12 organic acid and Ceylon cinnamon and its use wherein the organic acid is a carboxylic acid with acidic properties.
INTRODUCTION
Organic acids are widely used for different purposes in different combinations all over the world. It’s used as a disinfectant, as a food/feed preservation technique, as a growth promoter in production animals and many more. Organic acids have been successfully used as a growth promoter in pigs for more than 25 years and they have also been found to be effective in poultry production. From the use of organic acids in poultry and pigs, one can expect an improvement in performance similar to or better than that of antibiotic growth promoters, without the public health concern.
It has been observed that animals exposed to severe stress or when high performance is demanded, suffer from fatigue, diarrhea, resistance to feed intake, anemia, etc. when they only are fed standard feed. However, it is usually difficult to define what the causes for the observed problems are, and thus which additive to use. There are known numerous additives and feed supplements, but none have proved to solve all the above problems. Some additives are primarily intended for increased growth of the animal, while others claim to improve its health. Vitamin deficiencies might be part of the problem, but then one should understand why this occur even when feed is expected to contain sufficient amounts of vitamins.
Another observation in relation to present invention is the direct effect indicated on bacteria, virus and parasites. Especially in low-pH environments like stomach and abomasum(ruminants). The effects of organic acids on bacteria are known and the mechanism without being bound by the theory to relate to the non-dissociated organic acids obtained in an environment lower than pKa value of the respective organic acid».
Less is known about the direct effects of organic acids on viruses and parasites.
The present invention aims to solve one of the aforementioned problems and provides a composition comprising at least one Cl -Cl 2 organic acid and/or its salts
in combination with an iron component , Ceylon cinnamon and B-vitamins and wherein the organic acid is a carboxylic acid with acidic properties.
SUMMARY OF THE INVENTION
The present inventors have solved the above-mentioned need by providing in a first aspect a composition comprising at least one C1-C12 organic acid and/or its salt, an iron component, Ceylon cinnamon and B-vitamins in an amount from 0 to 200 mg/g dry weight of the composition and in amounts corresponding to at least that which theoretically can be consumed during metabolism of the COOH-groups present wherein the organic acid is a carboxylic acid with acidic properties according Bronsted definition of an acid which is a molecule that can donate a proton to another molecule.
In one embodiment according to the first aspect t he composition does not comprise L-carnitine.
In one embodiment according to the first aspect the organic acid is a C1-C8 and/or its salts wherein the total amount of the organic acids is in the range from 66 % (w/w) to 100% (w/w) by weight based on dry weight of the composition.
In one embodiment according to the first aspect the organic acid is a C1-C8 and/or its salts wherein the total amount of the organic acids is in the range from 66 % (w/w) to 99% (w/w) by weight based on dry weight of the composition, such as from 60% w/w to 95% w/w, such as from 75% w/w to 85% w/w.
In one embodiment according to the first aspect the organic acid is a fatty acid and/or its salts.
In one embodiment of the first aspect the salt is selected from a Sodium salt or a Potassium salt. In one embodiment according to the first aspect the organic acid is a Cl -Cl 2 fatty acid selected from short chain fatty acids, medium chain fatty acids and long chain fatty acids.
In one embodiment according to the first aspect the organic acid is a C1-C8 fatty acid.
Preferably the fatty acids are selected from short chained fatty acids (SCFA): formic acid (Cl), acetic acid (C2), Propionic acid (C3), Butyric acid (C4), Fumaric acid (C4); Medium-chained fatty acids (MCFA): Sorbic acid (C6), Caprylic acid (C8) and Long-chained fatty acids (LCFA): Lauric acid (C12).
In one embodiment according to the first aspect the at least one organic acid is formic acid and its salt.
In one embodiment according to the first aspect the iron component is ferrous fumarate.
In one embodiment according to the first aspect the fatty acids are selected from Caprylic acid (C8), Sorbic acid (C6), formic acid (Cl), Propionic acid (C3), Butyric acid (C4), acetic acid (C2) and any combinations thereof.
In one embodiment according to the first aspect the iron component is in an amount of 0 to 3.5% (w/w) by weight based on dry weight of the composition and the iron compound is preferably ferrous fumarate,
In one preferred embodiment the iron component is in an amount of 0.5 to 3.5% (w/w) by dry weight of the composition.
In a most preferred embodiment, the iron component is in an amount of 0.5 to 3.5% (w/w) by dry weight of the composition and the iron compound is ferrous fumarate.
In one embodiment according to the first aspect the composition of B-vitamins comprises 0-10 mg/g Bl, 0-25mg/g B2, 0-40 mg/g B3, 0-25 mg/g B5, 0-60 mg/g B6, 0-25 mg/g B7, 0-60 mg/g B9 and 0-250 pg/g B12 based on dry weight of the composition.
In one embodiment according to the first aspect the composition of B-vitamins comprises 2-10 mg/g Bl, 5-25mg/g B2, 10-25 mg/g B6, 0-25 mg/g B7, 0.1-60 mg/g B9 and 1-250 pg/g B12 based on dry weight of the composition.
In one embodiment according to the first aspect the composition of B-vitamins comprises 0.5-10 mg/g Bl, 0.5-25mg/g B2, B6 0.5-25 mg/g, 0-25 mg/g B7, 0.1-60 mg/g B9 and 10-250 pg/g B12 based on dry weight of the composition.
The amount of vitamins, B6, B9 and B 12 is in the range from 10-50mg/g dry weight of the composition and the amount of B9 and B12 should at least correspond to that which can be consumed during metabolism of the COOH-group of the carboxylic acids in particular in combination with formic acid and is salt.
In one embodiment according to the first aspect the Ceylon cinnamon is bark from Cinnamomum verum and is in an amount from 0.1 to 6% (w/w) by dry weight of the composition, preferably in an amount from 0.5 to 6% (w/w) by dry weight of the composition.
In one embodiment according to the first aspect the composition comprises the composition further comprises a desiccant in an amount of 0-6% (w/w) by dry weight of the composition, wherein the desiccant is selected from Silicone dioxide, Magnesium oxide or any combinations thereof.
In one embodiment according to the first aspect the composition comprises an antioxidant in an amount of 0-1% (w/w) by dry weight of the composition, wherein the antioxidant is preferably selected from Vitamin C, Vitamin E or any combinations thereof.
In one embodiment according to the first aspect the amount of salt and carboxylic acids will give a pH amounting to 2.0 to 7.5 when the composition is in form of a suspension.
In one embodiment according to the first aspect the amount of salt and carboxylic acids will give a pH amounting to 2.0 to 7.5 when the composition is dissolved in water.
In one embodiment according to the first aspect the amount of salt and carboxylic acids will give a pH amounting to 2.0 to 6.0 when the composition is in form of a suspension.
In one embodiment according to the first aspect the amount of salt and carboxylic acids will give a pH amounting to 2.0 to 6.0 when the composition is dissolved in water.
In one embodiment according to the first aspect the composition is a dietary supplement.
In one embodiment according to the first aspect the composition is a pharmaceutical composition.
In one embodiment according to the first aspect the composition is administrated in the state of powder, suspension, gel-suspension, supplemented directly to food or feed or is swallowed as a tablet or capsule.
In one embodiment according to the first aspect there is provided use of the dietary supplement for decreasing infectivity of a virus and/or a parasite in a subject .
In one embodiment according to the first aspect there is provided use of the dietary supplement for increasing virus and/or parasite resistance in a subject.
In one embodiment according to the first aspect there is provided a pharmaceutical composition for use in reducing the risk of and/or aid in treatment of a parasitic infection and/or viral infection in a subject.
In one embodiment of the first aspect the parasite is caused by a parasite with a hard-shelled life cycle stage where it forms a cyst and/or oocyst.
In one embodiment of the first aspect the parasite and/or the virus is comprised in the gastrointestinal system such as the alimentary tract and/or respiratory system of the subject.
In one embodiment of the first aspect the virus is selected from rotaviruses, adenoviruses, coronaviruses, Hepatitis A, Hepatitis E, Norovirus, Enterovirus and Poliovirus.
In one embodiment of the first aspect the parasite is selected from coccidiosis and giardiasis.
In one embodiment of the first aspect the fatty acids are selected from Sodium Caprylate, Potassium Sorbate, Sodium Formate, Sodium Butyrate, Sodium Acetate, Sodium Propionate and any combinations thereof wherein the composition is for use in preventing or treatment of a parasite comprised in the gastrointestinal system such as the alimentary tract and/or respiratory system of the subject.
In one embodiment of the first aspect the fatty acids are selected from Sodium Caprylate, Potassium Sorbate, Sodium Formate, Sodium Butyrate, Sodium Propionate and any combinations thereof wherein the composition is for use in preventing or treatment of a virus is comprised in the gastrointestinal system such as the alimentary tract and/or respiratory system of the subject.
In one embodiment of the first aspect the subject is an animal, a fish or a human.
In one embodiment of the first aspect the animal is selected from a ruminant, a horse, a race pigeon, a dog and a cat.
In one embodiment of the first aspect the composition is administrated as a daily dose in the range 25-500 mg dry weight / kg body weight or 0.5-10g per 20kg body weight
In one embodiment of the first aspect is administrated as a daily dose of about 1g per 20kg body weight for use in reducing the risk of a parasitic infection and/or viral infection in a subject.
In one embodiment of the first aspect the composition is administrated as a daily dose of 2g- 10g per 20kg body weight for use in treatment of or aid in treatment of a parasitic infection and/or viral infection in a subject.
In a second aspect there is provided a method decreasing infectivity of a virus and/or a parasite in a subject, comprising administering to the subject the composition according to the first aspect in a daily dose in the range from 25 to 500 mg dry weight / kg body weight of the subject.
In a third aspect there is provided a method for use in reducing the risk of and/or aid in treatment of a parasitic infection and/or viral infection in a subject comprising administering to the subject the composition according to the first aspect in a daily dose in the range from 25 to 500 mg dry weight / kg body weight of the subject.
In a fourth aspect there is provided a composition comprising one or more C1-C12 organic acid and/or its salt, an iron component, and B-vitamins for use in reducing the risk of and/or treatment of a parasitic infection and/or viral infection in a subject, wherein the B-vitamins is in an amount of 0-200 mg/g dry weight of the composition and the amount of B-vitamins corresponding to at least that which theoretically can be consumed during metabolism of the COOH-groups of the organic acid and wherein the organic acid is a carboxylic acid wherein the organic acid is a carboxylic acid with acidic properties according Bronsted definition of an acid which is a molecule that can donate a proton to another molecule.
The composition does not comprise D/L-carnitine. D/L-carnitine is derived from lysine and methionine and is involved in metabolism of fatty acids.
In one embodiment according to the fourth aspect the Cl -Cl 2 organic acid is a fatty acid and/or its salts.
In one embodiment of the fourth aspect the salt is selected from a Sodium salt or a Potassium salt
In one embodiment according to the fourth aspect the iron component is ferrous fumarate.
In one embodiment according to the fourth aspect the fatty acids are selected from Caprylic acid (C8), Sorbic acid (C6), formic acid (Cl), Propionic acid (C3), Butyric acid (C4), acetic acid (C2) and any combinations thereof.
In one embodiment according to the fourth aspect the at least one organic acid is formic acid and its salt.
In one embodiment according to the fourth aspect the fatty acids are selected from Sodium Caprylate, Potassium Sorbate, Sodium Formate, Sodium Butyrate, Sodium Propionate and any combinations thereof
In one embodiment according to the fourth aspect the composition is administration by inhalation or intranasal application in the state of powder, suspension, gelsuspension, supplemented directly to food/feed or is swallowed as a tablet or capsule.
In one embodiment according to the fourth aspect the parasite is caused by a parasite with a hard-shelled life cycle stage where it forms a cyst and/or oocyst.
In one embodiment according to the fourth aspect the parasite and/or the virus is comprised in the gastrointestinal system such as the alimentary tract and/or respiratory system of the subject.
In one embodiment according to the fourth aspect the virus is selected from rotaviruses, adenoviruses, coronaviruses, Hepatitis A, Hepatitis E, Norovirus, Enterovirus and Poliovirus.
In one embodiment according to the fourth aspect the parasite is selected from coccidiosis and giardiasis.
In one embodiment according to the fourth aspect the composition is an antiviral nasal spray.
DETAILED DESCRIPTION
Where a numerical limit or range is stated herein, the endpoints are included. Also, all values and sub ranges within a numerical limit or range are specifically included as if explicitly written out.
All methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, with suitable methods and materials being described herein. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will prevail.
In the following description, various examples and embodiments of the invention are set forth in order to provide the skilled person with a more thorough understanding of the invention. The specific details described in the context of the various embodiments and with reference to the attached drawings are not intended to be construed as limitations.
As mentioning above, the present invention provides a composition comprising at least one Cl -Cl 2 organic acid and/or its salts in combination with an iron component, Ceylon cinnamon and B-vitamins and wherein the organic acid is a carboxylic acid with acidic properties. The term C1-C12 organic acid as used herein refers to a fatty acid compound comprising from 1 carbon atom to 12 carbon atoms and having acidic properties. Acidic property is defined herein as a molecule that can donate a proton to another molecule according to Bronsted definition of acids, Bronsted, J. N. (1923). "Einige Bemerkungen uber den Begriff der Sauren und Basen" [Remarks on the concept of acids and bases], Recueil des Travaux Chimiques des Pays-Bas. 42 (8): 718-728
Preferably the Cl -Cl 2 carboxylic acids are selected from short chained fatty acids (SCFA): formic acid (Cl), acetic acid (C2), Propionic acid (C3), Butyric acid (C4), Fumaric acid (C4); Medium-chained fatty acids (MCFA): Sorbic acid (C6), Caprylic acid (C8) and Long-chained fatty acids (LCFA): Lauric acid (C12).
Most preferably at least one of the carboxylic acids is formic acid and its salt.
In one embodiment the fatty acids are selected from Caprylic acid (C8), Sorbic acid (C6), formic acid (Cl), Propionic acid (C3), Butyric acid (C4), acetic acid (C2) and any combinations thereof.
In one embodiment the present composition comprises Cinnamon, Sodium Caprylate, Potassium Sorbate, Sodium Formate, Sodium Butyrate, Sodium Propionate and any combinations thereof in a dietary supplement for increasing virus resistance in a subject wherein the virus is comprised in the gastrointestinal system such as the alimentary tract and/or respiratory system and wherein the virus is selected from rotaviruses, adenoviruses, coronaviruses, Hepatitis A, Hepatitis E, Norovirus, Enterovirus and Poliovirus.
In one embodiment the composition comprises Cinnamon, Sodium Caprylate, Potassium Sorbate, Sodium Formate, Sodium Butyrate, Sodium Propionate and any combinations thereof and in the form of a dietary supplement for decreasing infectivity of a virus and/or a parasite in a subject virus is comprised in the gastrointestinal system such as the alimentary tract and/or respiratory system and wherein the virus is selected from rotaviruses, adenoviruses, coronaviruses, Hepatitis A, Hepatitis E, Norovirus, Enterovirus and Poliovirus.
In one embodiment the composition comprises cinnamon, Sodium Caprylate, Potassium Sorbate, Iron Fumarate, Sodium Formate, Sodium Butyrate, Sodium Acetate, Sodium Propionate and any combinations thereof
Ceylon cinnamon as applied herein is bark from the Cinnamomum verum tree. Ceylon cinnamon from the Cinnamomum verum tree is often called true cinnamon and comprises lower amount of the toxin coumarin compared to Cassia cinnamon. The most produced cinnamon oil comes from Cassia cinnamon. Ceylon cinnamon is used herein.
Cinnamon oil can be made from either the bark or the leaves which will also give different ratios between the different essential oils. The composition of these two is very different in relation to the relationship between the different essential oils.
Cassia cinnamon contains approx. 250x more coumarin which in excessive doses can be toxic to the liver and kidneys. Too high doses of coumarin can also prevent
Vitamin K synthesis in the gut and "Vitamin K bleeding deficiency" The toxic effects of Coumarin probably affect most people.
There is a big difference in the ratio of essential oils between cinnamon bark and cinnamon leaves. Cinnamon oil from leaves often contains a lot of eugenol and very little cinnamaldehyde, while it is the other way around when using bark. We specify that we use the bark. Cinnamaldehyde has health-promoting effects by reducing the activity of important enzymes that trigger inflammatory reactions and allergic reactions.
The applicant and owners listed in this document is also the owner of European patent no. EP 1 691 626 Bl which is fully disclosed herein.
In particular table 3-5 of EP 1 691 626 Bl discloses recommended amounts of B6, B12, B9 and Fe for human and animals to be included in the composition of the present invention.
While the effects of organic acids on different pathogenic bacteria are widely described, less is known about their effects on viruses and parasites, especially in antiseptic use-cases. The infection route of many viruses does not involve fecal-oral route
However, for some viruses the fecal-oral rout seems to be more important than for others. Examples of viruses that are highly dependent on the fecal- oral route include Hepatitis A, Hepatitis E, Norovirus, Corona viruses (especially feline and equine coronaviruses), Enterovirus, Poliovirus and Rotavirus.
In viruses that depends largely on the fecal-oral route, recent findings may suggest that present invention decrease infectivity and aid in treatment and preventive treatment of infection.
Decrease infectivity as used herein is the reduction of a virus or parasites ability to infect a cell in a subject, i.e. increasing virus or parasitic resistance.
The effects of organic acids on different pathogenic parasites also seems to be lacking knowledge.
Some of the most common causes for protozoan parasitic diarrhoea in dogs and cats include coccidiosis and giardiasis. Among the coccidian parasites, Isospora and Cryptosporidium species are two of the most common causes of clinical manifestation of coccidiosis in dogs and cats. Other species may include Besnoitia, Toxoplasma, Hammondia and Sarcocystis. Coccidian parasites are known for their infectious oocysts with an obligate fecal-oral route of infection. Giardia parasites are another protozoan parasite with similar presentation of symptoms, however their life cycle do not include an oocyst. Their infectious stage is presented with cysts
shed in the feces of its host with an obligate fecal-oral route. Cysts and oocysts share many common properties including the hard-shelled structure that envelopes them. This makes them both highly resistant to chemical interference and environmental factors. Recent findings may suggest that present invention may treat or aid in the treatment of these.
Wild and farmed fish are also sometimes infected with viruses and parasites that has a hard-shelled life cycle stage. Examples include many coccidian parasites like Calyptospora, Goussia, Eimeria, Epieimeria, Cryptosporidium, Crystallospora, and Octosporella(fungi). However, coccidiosis in farmed fish in Norway seems to be a problem rather in lumpfish than in other farmed fish like atlantic salmon and rainbow trout. In Atlantic salmon and rainbow trout (salmonids), several viral diseases poses a threat to the industry. Examples include Cardiomyopathy syndrome, Pancreas disease, Infectious salmon anemia, infectious pancreatic necrosis, Heart and skeletal muscle inflammation, Viral hemorrhagic septicaemia, Infectious hematopoietic necrosis and Salmon gill pox virus.
There are many different antiprotozoal agents commercially available. This is a class of pharmaceuticals used in the treatment of protozoan diseases. Some of the most important protozoan diseases include amebiasis, giardiasis, cryptosporidiosis, microsporidiosis, malaria, babesiosis, trypanosomiasis, Chagas disease, leishmaniasis and toxoplasmosis. Currently, many of the treatments for these infections are limited by their toxicity and present invention may reducing the risk of and/or aid in the treatment of these.
Further details of the invention:
In one aspect it is described herein a combination of different components and its use against oocysts, cysts and other hard-shelled parasitic life cycle stages, and its use against coronaviruses, rotaviruses and other viruses with affinity towards alimentary tract and respiratory tract.
In a further aspect it is described a composition comprising as the basic component at least one C1-C12 organic acid and/or its salt, an iron component, Ceylon cinnamon and B-vitamins in 0-200mg/g dry matter and in amounts corresponding to at least that which theoretically can be consumed during metabolism of the COOH- groups present.
In a further aspect the composition also contains a desiccant and an antioxidant.
In a further aspect the composition may also be administered in a suspension with pH amounting to 2-7.5.
In a further aspect the composition contains 66-100% of one or more C1-C8 organic acids and/or its salts.
In a further aspect the composition is characterized by an iron component consisting of 0-3.5% (w/w) iron, and the component preferably used is ferrous fumarate.
In a further aspect the composition may also contain B-vitamins in amounts of 0- 200mg/g dry weight of the composition, wherein 0-10 mg/g Bl, 0-25mg/g B2, 0- 40mg/g B3, 0-25mg/g B5, 0-60mg/g B6, 0-25mg/g B7, 0-60mg/g B9 and 0-250pg/g B12.
In a further aspect the composition is characterized by 0-6% cinnamon, preferably from the plant Cinnamomum vera.
In a further aspect the composition is characterized by 0-6% w/w desiccant, preferably Silicone dioxide and/or Magnesium oxide.
In a further aspect the composition is characterized by 0-1% w/w antioxidant, preferably Vitamin C or Vitamin E.
In a further aspect the composition is used to treat or to aid in treatment against infection by parasites that contain hard-shelled life cycle stages like cysts and oocysts.
In a further aspect the composition is used to treat or aid in treatment against infection by viruses with an affinity towards gastrointestinal system and respiratory system like rotaviruses, adenoviruses, coronaviruses, etc.
In a further aspect the composition is administrated by a daily dosage which may be applied, is in the range 25-500mg/kg bodyweight or 0.5-10g per 20kg body weight.
In a further aspect the composition is administration by inhalation or intranasal application in the state of powder, suspension, gel-suspension, supplemented directly to food/feed or is swallowed as a tablet or capsule.
Having generally described this invention, a further understanding can be obtained by reference to certain specific examples. The examples illustrate the properties and effects of the composition according to the invention, and are provided herein for purposes of illustration only, and are not intended to be limiting.
The skilled person will understand that the main feature of the composition according to the invention comprises a basic component having at least one C1-C8 organic acid and/or its salt comprising at least formic acid, an iron component preferably iron fumarate, Ceylon cinnamon, and B6, B9 and B12-vitamins in the range from 0 to 200mg/g dry weight of the composition and in an amount corresponding to at least that which theoretically can be consumed during metabolism of the COOH-groups present and wherein the organic acid is a carboxylic acid having acidic properties and wherein the carboxylic acids are fatty
acids and wherein the total amount of organic acids is at least 66% (w/w) by weight based on dry weight of the composition. The skilled person will further understand that the composition in addition may comprise antioxidants and desiccants and further components depending on its intended use wherein these further components don’t change the effect of the composition.
EXAMPLES
Example 1
Mortality in fish may be caused by several factors including viruses and parasites as discussed above. A study conducted on 600 Atlantic salmon showed a 100% reduction in mortality rate. The study design was constructed as follows:
Feed A: Control carrier pellet with no additives
Feed B: Carrier pellet with 1.5% of present invention comprising Sodium diformate, Calcium butyrate and Sodium caprylate.
The fish were 40 grams and adapted to sea water at starting point.
Feeds were tested on four parallel tanks (total 12 tanks) with 50 fish in each tank (total 600 fish).
Results showed a 5% mortality rate in the control group after 9 and 10 weeks compared to zero mortality in group with carrier pellet. Growth and length were significantly improved within the first 5 weeks along with hepatic weight and intestinal tissue damage significantly reduced as well.
Example 2
A study investigating the effects of present composition comprising Sodium formate, Calcium formate, Iron formate and Sodium sorbate on gastric ulcers of the squamous mucosa in trotting racehorses went public. The study was performed as a randomized, double-blinded, single center study with stratified semi crossover design with breed as stratification factors. The horses were clinically and endoscopically examined prior to start and after 3 and 5 or 7 weeks of treatment. The ulcerations were scored in accordance with Equine Gastric Ulcer Council (EGUC) recommendations on a 5 point scale and on a 10 cm Visual Analogue Scale (VAS). The patients were responder-classified after 3 weeks. Responders in need of ulcer treatment were randomly allocated to 2 or 4 weeks of additional treatment. Non-responders to placebo were crossed to present invention. The 5-point EGUC
score and VAS recorded score was significantly reduced (P < 0.01) in both placebo and present invention groups after 3 weeks of treatment. From 3 weeks to the end of treatment the score was further significantly reduced in the group with present invention(P < 0.05). After the end of treatment, 78% in the group with present invention and 54.8% in the placebo group were classified as responders. The difference was significant (P = 0.04). There are several factors that may contribute to the manifestation of Equine gastric ulcer syndrome, but increased stress from gastrointestinal illness caused by viruses, parasites and bacteria are among them.
Example 3
Feedback from hundreds to thousands of customer clients who has purchased and used the present invention indicate that increased health, especially gastrointestinal health and symptoms of diarrhea is has been tested in indicative studies. The present invention contains salts of organic acids with established antibacterial effects, however results may suggest an even wider efficiency across viruses and parasites. Especially those involved in gastrointestinal illness.
Feedback from present invention against unstable stomach function in dogs: 91.6% were free from symptoms after 2-3 days.
Indicative study with present invention against unstable stomach function in horses: 100 %
Feedback after administering in calves was overwhelmingly positive as well. Customer satisfaction rate is estimated to 100% and included cases like
“1 month old calf with diarrhea was given present invention comprising Sodium formate, Calcium formate, Iron formate and Potassium sorbate mixed with milk and was free from symptoms the next day.“
“A calf with bloody diarrhea was given two meals supplemented with present invention and was immediately free from symptoms.”
“A newborn calf with diarrhea and decreased appetite was given present invention and was completely healthy after two days.”
Example 4
A field study on calves was conducted. A group of 39 calves equally divided by random into 7 groups where 3 of these groups were given present invention comprising Sodium formate, Calcium formate, Iron formate, Potassium sorbate and
Ceylon cinnamon and 4 of these groups were control groups. The results from this study showed that none of the calves fed with present invention developed diarrhea, 50% of the calves in the control group developed diarrhea, but were free from diarrhea within 3 days after being fed with present invention comprising Sodium formate, Calcium formate, Iron formate, Potassium sorbate and Ceylon cinnamon. The cause of diarrhea in these calves was unknown.
Example 5
Present invention was tested on a livestock farm that had previously been diagnosed with a rich establishment of Cryptosporidium pathogens. The calves in the barn recently started to show the same symptoms as before and the time of year also corresponded with cryptosporidium. It was regarded likely that Cryptosporidium was the cause of symptoms yet again. The first calf displaying symptoms of Cryptosporidium was treated with antibiotics, B-vitamins and present invention comprising Sodium formate, Calcium formate, Iron formate and Potassium formate and Ceylon cinnamon. Previous experience with this treatment method excluding present invention was rather bad, so usually supportive treatments like fluid pre- and probiotics and pH-regulators were tried first. The veterinarian expressed that the first calf displaying symptoms was free from symptoms unusually fast when treated with present invention and that the effect was not expected to be as fast and sufficient. The calves that previously shared pen with the sick calf also started to display symptoms of Cryptosporidium. These were treated with present invention without antibiotics or additional B-vitamins. They were free from symptoms shortly after treatment.
Example 6
A laboratory study was conducted where present invention was tested on a parasite and a virus. The parasite chosen was Cryptosporidium (Crypto) and the virus was Bovine Coronavirus (BCoV). These pathogens were tested with the use of human colorectal adenocarcinoma cells (HCT-8).
Results from BCoV:
Results from Cryptosporidium:
In the first two rows, we see no effect of the low pH as such, but in the last two rows, we see the effect when the present invention was added at similar pH levels.
The results from this setup shows how present invention is especially effective in low pH environment like stomach or abomasum or when present invention itself has a low pH.
Example 7
A laboratory study with the purpose to investigate whether Cryptosporidium parvum oocysts (the transmission stage of C. parvum) would have their ability to infect and replicate in a commonly used cell culture cell line (human colon adenocarcinoma; HCT- 8) affected following exposure to the present invention. Infection and replication within the cell line was determined by qPCR.
Oocysts were exposed to hypochlorite for 10 min on ice to remove bacterial contaminants and then washed twice in PBS before being resuspended in RPMI (a growth medium used in cell culture) at either pH 7 or at pH 3, the latter with or without (controls) different components from present invention. Single compounds were tested at concentrations of 5 mg/mL, and combinations with sodium formate at 2.5 mg/mL for each compound.
The experimental set-up was as shown in Figure 1 (below). The study was run in triplicate (three individual set-ups), with each set-up consisting of technical triplicates of each experimental procedure.
The copy number results obtained are shown in Table 1, and indicate the mean copy numbers of the qPCR target gene of C. parvum from the three replicate experiments. Higher numbers thus indicate greater numbers of parasites following infection and replication. Lower numbers indicate either reduced infection and/or reduced replication.
Table 1. Percentages of inhibition of Cryptosporidium parvum infection and/or replication following incubation with different components, as related to incubation in RPMI pH 3 in the absence of these compounds.
Compound % Control (RPMI3) % Inhibition
I (Iron Fumarate) 6.35 93.7
C (Cinnamon) 34.69 65.3
PS (Potassium Sorbate) 25.69 74.3
SA (Sodium Acetate) 36.76 63.2
SB (Sodium Butyrate) 19.64 80.4
SC (Sodium Caprylate) 1.39 98.6
SF (Sodium Formate) 1.77 98.2
SP (Sodium Propionate) 14.16 85.8
I+SF (Iron Fumarate + Sodium Formate) 1.56 98.4
C+SF (Cinnamon + Sodium Formate) 0.75 99.3
PS+SF (Potassium Sorbate + Sodium Formate) 3.22 96.8
SUBSTITUTE SHEET (RULE 26)
SA+SF (Sodium Acetate + Sodium Formate) 1.82 98.2
SB+SF (Sodium Butyrate + Sodium Formate) 2.66 97.3
SC+SF (Sodium Caprylate + Sodium Formate) 0.65 99.4
SP+SF (Sodium Propionate + Sodium Formate) 4.05 96.0
It is clear, that exposure of oocysts to RPMI at pH 3 promotes their cultivation (infection and/or replication) as the copy number is significantly greater than that obtained from oocysts incubated in RPMI at pH 7 (without any organic acid components) prior to inoculation into the cell culture. Although incubation of the oocysts in RPMI at pH 7 resulted in more successful cultivation than that obtained from oocysts simply held in PBS, this was not significantly different. The reasons for the increase in cultivation success from incubation in RPMI at pH3 compared with incubation in RPMI at pH7 can only be speculated upon, as they were not investigated here. However, they may indicate that the low pH stimulates excystation and free sporozoites are more available and susceptible to other factors in the media (such as those in the present invention) or other external pressures. In general, exposure of Cryptosporidium oocysts to sodium formate in RPMI at pH 3, either alone or in combination with other components of present invention, had a negative effect on their ability to infect and/or replicate in vitro. Other components that, alone, clearly had a negative effect were Sodium Caprylate and Iron Fumarate. The other components alone (cinnamon, Potassium Sorbate, Sodium Acetate, Sodium Butyrate, and Sodium Propionate) showed lower copy numbers than the control oocysts in RPMI at pH 3 (reduced 6 infection and or replication), but not significantly different to values obtained from oocysts in PBS or incubated in RPMI at pH 7.
Example 8
A laboratory study was conducted to test the effect of powder from cinnamon bark from Ceylon cinnamon on the infectivity of a virus (Experiment A and B) and to test the effect on the infectivity of a virus, using a combination of the following organic acids: Sodium format, Calcium format, iron fumarate and potassium sorbate (experiment C and D). The virus used for the experiments was Bovine Coronavirus (BCoV). The infectivity of the virus was tested with the use of human colorectal adenocarcinoma cells (HCT-8) and the infectivity of the virus was evaluated using microscope.
Experiment A
Cinnamon, pH 7 % Infectivity virus
3,75 mg/ml 42
0 mg/ml 100
Experiment B
Cinnamon, pH3 % Infectivity virus 3,75 mg/ml 1 0 mg/ml 100
Experiment C
Salts, pH7 % Infectivity virus 3,75 mg/ml 88
9,4 mg/ml 54
0 mg/ml 100
Experiment D
Salts, pH3 % Infectivity virus 3,75 mg/ml 10
9,4 mg/ml 16
0 mg/ml 100
The results from this setup shows how present invention is especially effective in low pH environment like stomach or abomasum or when present invention itself has a low pH. This effect, however without being bound by the theory, is likely due to the properties of non-dissociated organic acids obtained when suspended in an environment lower than their respective pKa values of the organic acids. pH 3 in this study is lower than pKA values of the 3 organic acids included in this example
Claims
1. A composition comprising at least one C1-C12 organic acid and/or its salt, an iron component, Ceylon cinnamon and B-vitamins in an amount from 0 to 200mg/g dry weight of the composition and in amounts corresponding to at least that which theoretically can be consumed during metabolism of the COOH-groups present wherein the organic acid is a carboxylic acid with acidic properties according Bronsted definition of an acid which is a molecule that can donate a proton to another molecule.
2. The composition according to claim 1, wherein the organic acid is a C1-C8 and/or its salts in an amount of 66 % (w/w) to 100% (w/w) by weight based on dry weight of the composition.
3. The composition according to claim 1 or claim 2, wherein the organic acid is a fatty acid.
4. The composition according to any one of claims 1 to 3, wherein the fatty acids are selected from Caprylic acid (C8), Sorbic acid (C6), formic acid (Cl), Propionic acid (C3), Butyric acid (C4), acetic acid (C2) and any combinations thereof.
5. The composition for use according to any one of claims 1 to 4, wherein the iron component is in an amount of 0 to 3.5% (w/w) by weight based on dry weight of the composition and the iron compound is preferably ferrous fumarate.
6. The composition according to any one of claims 1 to 5, wherein the composition comprises 0-10 mg/g Bl, 0-25mg/g B2, 0-40 mg/g B3, 0-25 mg/g B5, 0-60 mg/g B6, 0-25 mg/g B7, 0-60 mg/g B9 and 0-250 pg/g B12 dry weight of the composition.
7. The composition according to any one of claims 1 to 6, wherein the Ceylon cinnamon is bark from Cinnamomum verum and is in an amount from 0.001 to 6% (w/w) by weight based on dry weight of the composition, preferably in an amount from 0.5 to 6% (w/w) by weight based on dry weight of the composition, and more preferably in an amount from 1 to 6% (w/w) by weight based on total dry weight of the composition.
8. The composition according to any one of claims 1 to 7, wherein the composition comprises a desiccant in an amount of 0-6% (w/w) by weight based on
dry weight of the composition, wherein the desiccant is selected from Silicone dioxide, Magnesium oxide or any combinations thereof.
9. The composition according to any one of claims 1 to 8, characterized in that the composition comprises an antioxidant in an amount of 0-1% (w/w) by weight based on dry weight of the composition , wherein the antioxidant is preferably selected from Vitamin C, Vitamin E or any combinations thereof.
10. The composition according to any one of claims 1 to 9, wherein the amount of salt and carboxylic acids will give a pH amounting to 2.0 to 7.5 when the composition is in form of a suspension.
11. A dietary supplement comprising the composition according to any one of claims 1 to 10.
12. Pharmaceutical composition comprising the composition according to any one of claims 1 to 10.
13. The dietary supplement according to claim 11, wherein the supplement is administrated in the state of powder, suspension, gel-suspension, supplemented directly to food or feed or is swallowed as a tablet or capsule.
14. Use of the dietary supplement according to claim 13 for decreasing infectivity of a virus and/or a parasite in a subject.
15. Use of the dietary supplement according to claim 13 for increasing virus and/or parasite resistance in a subject.
16. The pharmaceutical composition according to claim 12 for use in reducing risk of and/or in treatment of gastro intestinal illnesses such as gastric ulcer and diarrhea in a subject, caused by a parasitic infection and/or viral infection.
17. The pharmaceutical composition according to claim 16 for use, wherein the parasite is caused by a parasite with a hard-shelled life cycle stage where it forms a cyst and/or oocyst.
18. The pharmaceutical composition according to claim 16 for use, wherein the parasite and the virus is comprised in the gastrointestinal system such as the alimentary tract and/or respiratory system of the subject.
19. The pharmaceutical composition according to claim 16 for use, wherein the virus is selected from rotaviruses, adenoviruses, coronaviruses, Hepatitis A, Hepatitis E, Norovirus, Enterovirus and Poliovirus.
20. The pharmaceutical composition according to any one of claims 16 for use, wherein the parasite is selected from coccidiosis and giardiasis.
21. The pharmaceutical composition according to claim 16 for use, wherein the subject is an animal, a fish or a human.
22. The pharmaceutical composition according to claim 21 for use, wherein the animal is selected from a ruminant, a horse, a race pigeon, a dog and a cat.
23. The pharmaceutical composition according to claim 12, wherein the pharmaceutical composition is administrated as a daily dose in the range from 25 to 500 mg dry weight / kg body weight.
24. A method decreasing infectivity of a virus and/or a parasite in a subject, comprising administering to the subject the composition according to any one of claims 1 to 10 as a daily dose in the range from 25 to 500 mg dry weight / kg body weight of the subject.
25. A method for use in reducing the risk of and/or in treatment of a parasitic infection and/or viral infection in a subject comprising administering to the subject the composition according to any one of claims 1 to 10 a daily dose in the range from 25 to 500 mg dry weight / kg body weight of the subject.
26. A composition comprising one or more C1-C12 organic acid and/or its salt, an iron component, and B-vitamins for use in reducing the risk of and/or treatment of a parasitic infection and/or viral infection in a subject, wherein the B-vitamins is in an amount of 0-200 mg/g dry weight of the composition and the amount of B- vitamins corresponding to at least that which theoretically can be consumed during metabolism of the COOH-groups of the organic acid and wherein the organic acid is a carboxylic acid wherein the organic acid is a carboxylic acid with acidic properties according Bronsted definition of an acid which is a molecule that can donate a proton to another molecule.
27. The composition according to claim 26, wherein the fatty acids are selected from Caprylic acid (C8), Sorbic acid (C6), formic acid (Cl), Propionic acid (C3), Butyric acid (C4), acetic acid (C2) and any combinations thereof.
28. Sodium Caprylate, Potassium Sorbate, Sodium Formate, Sodium Butyrate, Sodium Propionate and any combinations thereof.
29. The composition according to any one of claims 26, wherein the composition is administration by inhalation or intranasal application in the state of powder, suspension, gel-suspension, supplemented directly to food/feed or is swallowed as a tablet or capsule.
30. The composition according to any one of claims 26 to 28 wherein the composition is an antiviral nasal spray.
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US5858356A (en) * | 1995-12-21 | 1999-01-12 | Abbott Laboratories | Lactobacillus acidophilus to inhibit cryptosporidiosis in mammals |
EP1691626A1 (en) | 2003-12-05 | 2006-08-23 | Pigeon Vitality AS | Food and feed supplement and its use |
US10478493B2 (en) * | 2015-08-31 | 2019-11-19 | Stolle Milk Biologics, Inc. | Method of treating protozoal gastrointestinal disorders in immunocompromised patients |
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2022
- 2022-03-08 WO PCT/EP2022/055947 patent/WO2023169664A1/en unknown
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- 2023-03-08 WO PCT/EP2023/055944 patent/WO2023170183A1/en unknown
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US5858356A (en) * | 1995-12-21 | 1999-01-12 | Abbott Laboratories | Lactobacillus acidophilus to inhibit cryptosporidiosis in mammals |
EP1691626A1 (en) | 2003-12-05 | 2006-08-23 | Pigeon Vitality AS | Food and feed supplement and its use |
EP1691626B1 (en) | 2003-12-05 | 2009-02-25 | Vitality Innovation AS | Food and feed supplement and its use |
US10478493B2 (en) * | 2015-08-31 | 2019-11-19 | Stolle Milk Biologics, Inc. | Method of treating protozoal gastrointestinal disorders in immunocompromised patients |
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ALMORADIE ALAIN MAE . ET AL: "Cryptosporicidal Activity of Plant Extracts against Cryptosporidium parvum and Cryptosporidium hominis Thyroid genetic disorders View project Tissue Engineering and molecular detection View project", ASIAN JOURNAL OF PHARMACOGNOSY, vol. 2, no. 3, 1 January 2018 (2018-01-01), pages 22 - 31, XP055965431, DOI: 10.13140/RG.2.2.17527.98728 * |
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GÓMEZ-GARCÍA MANUEL ET AL: "In vitro Assessment of Antiviral Effect of Natural Compounds on Porcine Epidemic Diarrhea Coronavirus", FRONTIERS IN VETERINARY SCIENCE, vol. 8, 29 March 2021 (2021-03-29), XP055965454, DOI: 10.3389/fvets.2021.652000 * |
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