WO2004087176A1 - A composition (rucd) for protecting and/or repairing dna from oxidative damages and a method thereof - Google Patents
A composition (rucd) for protecting and/or repairing dna from oxidative damages and a method thereof Download PDFInfo
- Publication number
- WO2004087176A1 WO2004087176A1 PCT/IN2003/000125 IN0300125W WO2004087176A1 WO 2004087176 A1 WO2004087176 A1 WO 2004087176A1 IN 0300125 W IN0300125 W IN 0300125W WO 2004087176 A1 WO2004087176 A1 WO 2004087176A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- composition
- dna
- acid
- cow
- rcud
- Prior art date
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Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
- A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
- A61K31/19—Carboxylic acids, e.g. valproic acid
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
- A61K31/19—Carboxylic acids, e.g. valproic acid
- A61K31/192—Carboxylic acids, e.g. valproic acid having aromatic groups, e.g. sulindac, 2-aryl-propionic acids, ethacrynic acid
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K33/00—Medicinal preparations containing inorganic active ingredients
- A61K33/02—Ammonia; Compounds thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/22—Urine; Urinary tract, e.g. kidney or bladder; Intraglomerular mesangial cells; Renal mesenchymal cells; Adrenal gland
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
- A61P39/06—Free radical scavengers or antioxidants
Definitions
- a composition useful for protecting and/or repairing DNA from oxidative damages said composition comprising redistilled cow's urine distillate (RCUD) having components benzoic acid, and hexanoic acid, with ammonia content of the composition ranging between 5-15mg/L, and optionally along with anti-oxidants; and a method of protecting and/or repairing DNA from oxidative damages using composition of claim 1, said method comprising steps of estimating the amount of folded DNA in a sample, mixing the said composition to the said DNA either before or after the exposure of the DNA to the oxidatively DNA-damaging agent, and determining percentage folded DNA in the mixture showing protection and/or repair of DNA from oxidative damages.
- RCUD redistilled cow's urine distillate
- Cow's urine is one of the ingredient of Panchagavya. According to Susrut, Cow's urine has several medicinal properties (45/221, pp-61, 72, 220, 221). Its properties are also described by Charak (Solka-100 pp. 72). Cow's urine is extensively used in Ayurveda for purifying certain materials which otherwise possess toxic properties (pp-73).
- the literature and scripture does not mention urine or its distillate as a protector of cells from DNA damage and chromosomal aberration.
- Kamdhenu Ark cow urine distillate prepared and marketed by the Govigyan Anusandhan Kendra (GVAK), Nagpur (India). This is the urine distillate suggested for oral ingestion to improve the general health and to control weight gain, oedema, stomach ache, indigestion, skin disease, cardiac problem etc. It is stated in the production process of Kamdlienu Ark that the cow's urine should be fresh as well as free of ammonia. Experiments carried out in NEERI revealed that storage of cow's urine results in formation of ammonia as could be seen from the following Table 1. Table 1 : Ammonia concentration in cows urine
- the main object of the present invention is to develop a natural composition for preventing and rectifying DNA damage.
- Another main object of he present invention is to develop a method of preventing and rectifying DNA damage.
- Yet another object of the present invention is to develop a process of preparing RCUD.
- the main objective is to provide new uses of Kamdhenu Ark as a protector of DNA damage.
- a composition useful for protecting and or repairing DNA from oxidative damages said composition comprising redistilled cow's urine distillate (RCUD) having components benzoic acid, and hexanoic acid, with ammonia content of the composition ranging between 5-15mg/L, and optionally along with anti-oxidants; and a method of protecting and/or repairing DNA from oxidative damages using composition of claim 1, said method comprising steps of estimating the amount of folded DNA in a sample, mixing the said composition to the said DNA either before or after the exposure of the DNA to the oxidatively DNA-damaging agent, and determining percentage folded DNA in the mixture showing protection and/or repair of DNA from oxidative damages.
- RCUD cow's urine distillate
- a composition useful for protecting and/or repairing DNA from oxidative damages said composition comprising redistilled cow's urine distillate (RCUD) having components benzoic acid, and hexanoic acid, with ammonia content of the composition ranging between 5-15mg/L, and optionally along with anti-oxidants; and a method of protecting and/or repairing DNA from oxidative damages using composition of claim 1, said method comprising steps of estimating the amount of folded DNA in a sample, mixing the said composition to the said DNA either before or after the exposure of the DNA to the oxidatively DNA-damaging agent, and determining percentage folded DNA in the mixture showing protection and/or repair of DNA from oxidative damages.
- RCUD redistilled cow's urine distillate
- composition useful for protecting and/or repairing DNA from oxidative damages comprising redistilled cow's urine distillate (RCUD) having components benzoic acid, and hexanoic acid, with ammonia content of the composition ranging between 5-15mg/L, and optionally along with anti-oxidants.
- RCUD redistilled cow's urine distillate
- the RCUD is water-soluble. In still another embodiment of the present invention, wherein the RCUD shows specific gravity of about 1.006.
- the RCUD shows ammonical nitrogen (NH 3 N) of concentration ranging between 5-15 mg/1.
- the RCUD contains volatile fatty acids in the range of 1000 to 3000 mg/1.
- the antioxidants are selected from a group comprising volatile acids, quinoline, and hydrocarbon derivatives.
- volatile fatty acids are derivatives of acetic acid, propionic acid, and butyric acid.
- the derivatives of the volatile fatty acids are : Acetic acid- 2propenyl ester; Acetic acid mercapto methyl ester; 2 - Butenedinitrile, Fumanonitrile; Propioinc acid, 2,2,3-trichloro; Propioinc acid-3-chloro; 2- Pentenoic acid-3 methyl; Iso valeric acid; Butanoic acid, 3 hydroxy, ethyl ether; Benzoic acid , 4methyl 2-trimethyl; Acetic acid[o-(triethyl siloxy) phenyl tri ethyl silyl; Propionic acid, 3 phenyl (cinnamic acid); Acetic acid mercapto; 1-H indole-3-ol-acetate; Acetic acid phenyl ester; Quinoline, 3 -methyl.
- quinoline is selected from a group comprising quinoline 3-methylthi and quinolines 3-methyl.
- composition smells like urine.
- composition is used as medicinal composition.
- composition is used for analysis for genetic material.
- a method of protecting and/or repairing DNA from oxidative damages using composition of claim 1 comprising steps of; o estimating the amount of folded DNA in a sample, o mixing the said composition to the said DNA either before or after the exposure of the DNA to the oxidatively DNA-damaging agent, and
- DNA is damaged with compounds selected from a group comprising Actinomycin D, Manganese Oxide (MnO ), and Hydrogen Peroxide (H H 2 ).
- composition enhances anti-genotoxic effect by about 150-250 folds.
- a method of removing ammonia from cow urine distillate to obtain redistilled cow's urine distillate comprising steps of re-distillating the cow's urine distillate using Orthophosphoric acid of concentration ranging between 82% to 88%, and obtaining redistilled cow's urine distillate (RCUD).
- a pharmaceutical composition comprising antioxidants containing cow urine distillate in an amount effective' to protect DNA damage is disclosed.
- the antioxidants can be volatile fatty acid.
- the invention related to a noble use of redistilled low ammonia containing cow's urine distillate for DNA damage protection.
- DNA one of the most critical cellular targets for hazardous chemicals and wastes, may get damaged following exposure by alteration of bases of disruption of sugar phosphate backbone.
- the present invention has direct implication in facilitating the damage repair caused by DNA damaging chemical such as hydrogen peroxide (H 2 O 2 ), actinomycin-D and manganese-dioxide (MnO 2 ).
- the ammonia originates from urea present in cow's urine following enzymatic hydrolysis by the enzyme urease present in number of microorganisms. Though ammonia is toxic to human health, the dose at which Kamdhenu Ark is consumed (5 ml at a time) is not likely to exert any adverse health effect or healthy individuals. However, its removal from the Ark is desirable before human consumption at least for the sensitive population. Therefore Kamdhenu Ark procured from GVAK, Nagpur was treated with orthophosphoric acid following which distillation was again carried out. The redistillation process require addition of 10-25 ml of (85%) Orthophosphoric acid (AR grade) to 400 ml of K-Ark to bring the pH to less than 2 which will immobilize ammonia in the residue.
- Orthophosphoric acid AR grade
- the redistillation was performed at around 100°C and the distillate was tested for ammonia. It was observed that the ammonia level in the distillate varied between 5-15 mg/1. The removal of ammonia resulted in disappearance of the manifestation of cytotoxicity, which was observed with ammonia containing K-Ark prior to redistillation.
- the redistilled Kamdhenu-Ark (RCUD) with an ammonia level varying between 5 and 15 mg/1 was used for subsequent experiments.
- the present invention was the result of planned experiment which demonstrated the property of Kamdhenu-Ark as the protector of DNA damage to cells exposed to DNA damaging chemicals.
- Cow's urine one of the components of Panchagavya, is considered to cause weight loss reversal of certain cardiac problems, indigestion, stomach ache, oedema, etc.
- its role in preventing DNA damage has not been mentioned anywhere in the literature and scriptures. Therefore the applicant considered worthwhile to determine the efficiency of redistilled Kamdhenu Ark to protect DNA damage in cells exposed to DNA damaging chemicals in vitro.
- the applicants also analyzed the redistilled Kamdhenu Ark using gas chromotography-mass spectrography (GC-MS) and identified the components which are likely to act as anti-oxidants to behave like other known antioxidants which are known to protect DNA damage.
- the invention relates new uses of cow's urine distillate as a protectant of cellular DNA damaging chemicals such as actinomycin-D, hydrogen peroxide and manganese dioxide.
- the compounds identified in redistilled cow's urine acts as antioxidants which are known to protect cells from DNA damage.
- PMNs polymorphonuclear leucocytes
- DNA damage caused by the well known DNA damaging chemical hydrogen peroxide was statistically significant; (P ⁇ 0.001).Pretreatment of polymorphonuclear leucocytes (PMNs) with 60 ⁇ l of redistilled cow's urine distillate, did not result in statistically significant DNA damage in cells exposed to H 2 O 2 and redistilled cow's showing that redistilled cow urine distillate can protect cells from DNA damage.
- PMNs polymorphonuclear leucocytes
- the DNA damage caused by the well known DNA damaging chemical manganese dioxide was statistically significant (P ⁇ 0.001).
- Polymorphonuclear leucocytes (PMNs) were exposed simultaneously to MnO 2 and 60 ⁇ l of redistilled cow urine distillate, no statistically significant DNA damage was observed.
- Table 5 DNA damage induced by hydrogen peroxide (H2O 2 ) and protection by redistilled cow's urine distillate (l ⁇ l concentration).
- This invention is the result of a search for antioxidants of natural origin with a property to protect DNA damage in cells exposed to DNA damaging chemicals.
- of redistilled cow's urine distillate was experimented as a positive agent from DNA damaging chemicals.
- the redistilled cow's urine distillate was found to contain antioxidants such as 1) benzoic acid, hexanoic acid. In still another embodiment the redistilled cow's urine distillate was used as a positive agent for DNA damage caused by actinomycin D. In an embodiment, the redistilled cow's urine distillate was found to contain antioxidant such as 1) benzoic acid, hexanoic acid. In still another embodiment, the redistilled cow's urine distillate was used as a protection agent for DNA damage caused by H 2 O 2 In an embodiment, the redistilled cow's urine distillate was found to contain antioxidant. h still another embodiment the redistilled cow's urine distillate was used as a positive agent for DNA damage caused by MnO 2
- the effect of redistilled cow's urine distillate as a protective agent for DNA damaging chemical was studied after pre, simultaneous and post treatment of PMN's prior to exposure to DNA damaging chemicals.
- the Redistilled Cow Urine Distillate (RCUD) at a volume of 60 ⁇ l could result in providing protection to PMN's following exposure to actinomycin D.
- the Redistilled Cow Urine Distillate at a volume of 60 ⁇ l could result in providing protection to PMN's following exposure to Mn0 2 .
- Redistilled Cow Urine Distillate at a volume of 60 ⁇ l could result in providing protection to PMN's following exposure to H 2 O .
- Redistilled Cow Urine Distillate at a volume of 1 ⁇ l could result in providing protection to PMN's following exposure to H 2 O 2 .
- the tubes (control and three experimental) are mixed thoroughly and the mixture was equally distributed in the volume of 0.2 ml in 3 tubes designated as B, P and T, then 200 ⁇ l of solution C was added.
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- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Epidemiology (AREA)
- Cell Biology (AREA)
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- Toxicology (AREA)
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- Developmental Biology & Embryology (AREA)
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- Chemical Kinetics & Catalysis (AREA)
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- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
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- Inorganic Chemistry (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
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Abstract
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Priority Applications (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
AU2003226643A AU2003226643A1 (en) | 2003-03-31 | 2003-03-31 | A composition (rucd) for protecting and/or repairing dna from oxidative damages and a method thereof |
CNB038263750A CN100475221C (en) | 2003-03-31 | 2003-03-31 | A composition (RCUD) for protecting and/or repairing DNA from oxidative damages and a method thereof |
PCT/IN2003/000125 WO2004087176A1 (en) | 2003-03-31 | 2003-03-31 | A composition (rucd) for protecting and/or repairing dna from oxidative damages and a method thereof |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
PCT/IN2003/000125 WO2004087176A1 (en) | 2003-03-31 | 2003-03-31 | A composition (rucd) for protecting and/or repairing dna from oxidative damages and a method thereof |
Publications (1)
Publication Number | Publication Date |
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WO2004087176A1 true WO2004087176A1 (en) | 2004-10-14 |
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Application Number | Title | Priority Date | Filing Date |
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PCT/IN2003/000125 WO2004087176A1 (en) | 2003-03-31 | 2003-03-31 | A composition (rucd) for protecting and/or repairing dna from oxidative damages and a method thereof |
Country Status (3)
Country | Link |
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CN (1) | CN100475221C (en) |
AU (1) | AU2003226643A1 (en) |
WO (1) | WO2004087176A1 (en) |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2002032436A1 (en) * | 2000-10-20 | 2002-04-25 | Council Of Scientific And Industrial Research | Use of bioactive fraction from cow urine distillate ('go mutra') as a bio-enhancer of anti-infective, anti-cancer agents and nutrients |
-
2003
- 2003-03-31 CN CNB038263750A patent/CN100475221C/en not_active Expired - Fee Related
- 2003-03-31 AU AU2003226643A patent/AU2003226643A1/en not_active Abandoned
- 2003-03-31 WO PCT/IN2003/000125 patent/WO2004087176A1/en not_active Application Discontinuation
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2002032436A1 (en) * | 2000-10-20 | 2002-04-25 | Council Of Scientific And Industrial Research | Use of bioactive fraction from cow urine distillate ('go mutra') as a bio-enhancer of anti-infective, anti-cancer agents and nutrients |
Also Published As
Publication number | Publication date |
---|---|
CN100475221C (en) | 2009-04-08 |
CN1771045A (en) | 2006-05-10 |
AU2003226643A1 (en) | 2004-10-25 |
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