WO2021220124A1 - A pharmaceutical composition and method of treatment using l-carnosine and glutathione, its precursors or derivatives thereof - Google Patents

A pharmaceutical composition and method of treatment using l-carnosine and glutathione, its precursors or derivatives thereof Download PDF

Info

Publication number
WO2021220124A1
WO2021220124A1 PCT/IB2021/053397 IB2021053397W WO2021220124A1 WO 2021220124 A1 WO2021220124 A1 WO 2021220124A1 IB 2021053397 W IB2021053397 W IB 2021053397W WO 2021220124 A1 WO2021220124 A1 WO 2021220124A1
Authority
WO
WIPO (PCT)
Prior art keywords
derivatives
glutathione
pharmaceutically acceptable
acceptable salts
carnosine
Prior art date
Application number
PCT/IB2021/053397
Other languages
French (fr)
Inventor
Shyam Sunder Singh
Original Assignee
Shyam Sunder Singh
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Shyam Sunder Singh filed Critical Shyam Sunder Singh
Publication of WO2021220124A1 publication Critical patent/WO2021220124A1/en

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/02—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link
    • C07K5/0202—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link containing the structure -NH-X-X-C(=0)-, X being an optionally substituted carbon atom or a heteroatom, e.g. beta-amino acids
    • 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
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/4164—1,3-Diazoles
    • A61K31/4172—Imidazole-alkanecarboxylic acids, e.g. histidine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
    • A61K38/06—Tripeptides
    • A61K38/063—Glutathione
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/08—Tripeptides
    • C07K5/0802—Tripeptides with the first amino acid being neutral
    • C07K5/0804—Tripeptides with the first amino acid being neutral and aliphatic
    • C07K5/0806—Tripeptides with the first amino acid being neutral and aliphatic the side chain containing 0 or 1 carbon atoms, i.e. Gly, Ala

Definitions

  • the present invention relates to pharmaceutical composition comprising L- Carnosine or pharmaceutically acceptable salts or derivatives thereof, and Glutathione, its precursors or pharmaceutically acceptable salts or derivatives thereof.
  • the present invention also relates to method of treating disease or disorder in patient in need of such therapy, wherein treatment comprises administration of L-Carnosine or pharmaceutically acceptable salts or derivatives thereof, and Glutathione, its precursors or pharmaceutically acceptable salts or derivatives thereof, in same or different compositions to humans or animal.
  • Oxidative stress, ion balance and pH balance are some the most critical factors involved in the major acute and chronic physiological imbalance.
  • the depletion of powerful antioxidant renders cells particularly vulnerable to oxidative stress.
  • the resulting damage is the key step in the onset and progression of many disease states.
  • endogenous and exogenous compound are known to protect against oxidative stress. These endogenous and exogenous compound are combined with other therapies like antibiotics, anti-cancer drugs to achieved combined desired therapy goals.
  • L-Carnosine is used as protective agent for many therapies.
  • Glutathione is endogenous agent involved in anti-aging and other biochemical pathways providing shielding effect upon administration. While both these compounds are known to be administrated individual for their respective mechanism of action, individual administration has a limitation of affecting only their respective physiological involvement.
  • composition comprising L-Carnosine or pharmaceutically acceptable salts or derivatives thereof, and Glutathione, its precursors or pharmaceutically acceptable salts or derivatives thereof are not known.
  • Figure 1 Effect of combination of L- camosine and S-acetyl-L-glutathione on H2O2 scavenging activity.
  • Figure 3 Effect of 1-camosine and s-acetyl -1-glutathione on MDA levels in kidney tissues.
  • Figure 4 Effect of 1-camosine and s-acetyl-l-glutathione on catalase levels in kidney tissue.
  • Figure 5 Effect of 1-camosine and s-acetyl-l-glutathione on SOD levels in kidney tissues.
  • the present invention relates to pharmaceutical composition
  • pharmaceutical composition comprising L- Carnosine or pharmaceutically acceptable salts or derivatives thereof, and Glutathione, its precursors or pharmaceutically acceptable salts or derivatives thereof.
  • the active ingredient as per present invention are used in therapeutically effective amount.
  • “Therapeutically effective amount” or “effective amount” refers to the amount of a pharmaceutically active agent when administered to a patient, is sufficient to affect such treatment for the disease.
  • the therapeutically effective amount will vary depending on the disease and its severity, and the age, weight, and other conditions of the patient to be treated.
  • compositions refers to any composition for administration to human or animal by any route and includes but are not limited to immediate release, delayed release, extended release and pulsed-release.
  • the present invention relates to a pharmaceutical composition
  • a pharmaceutical composition comprising a. therapeutically effective amount of L-Carnosine or pharmaceutically acceptable salts or derivatives thereof, and b. therapeutically effective amount of Glutathione, its precursors or pharmaceutically acceptable salts or derivatives thereof.
  • the present invention relates to a pharmaceutical composition consisting of a. therapeutically effective amount of L-Carnosine or pharmaceutically acceptable salts or derivatives thereof, and b. therapeutically effective amount of Glutathione, its precursors or pharmaceutically acceptable salts or derivatives thereof c. one or more suitable inactive ingredients.
  • the present invention relates to a pharmaceutical composition
  • a pharmaceutical composition comprising c. therapeutically effective amount of L-Carnosine or pharmaceutically acceptable salts or derivatives thereof, and d. therapeutically effective amount of S-Acetyl Glutathione or pharmaceutically acceptable salts or derivatives thereof.
  • the present invention relates to a pharmaceutical composition consisting of d. therapeutically effective amount of L-Carnosine or pharmaceutically acceptable salts or derivatives thereof, and e. therapeutically effective amount of S-Acetyl Glutathione or pharmaceutically acceptable salts or derivatives thereof f. one or more suitable inactive ingredients.
  • the present invention relates to a pharmaceutical comprising a. L-Carnosine or pharmaceutically acceptable salts or derivatives thereof in an amount equivalent to 50mg to 400mg of L-Carnosine, and b. S-Acetyl Glutathione or pharmaceutically acceptable salts or derivatives thereof in an amount equivalent to lOOmg to 600mg of
  • the present invention relates to an oral pharmaceutical composition
  • an oral pharmaceutical composition comprising a. L-Carnosine in an amount between 50mg to 400mg, and b. S- Acetyl Glutathione in an amount between lOOmg to 600mg.
  • the present invention relates to an oral pharmaceutical composition
  • an oral pharmaceutical composition comprising a. L-Carnosine in an amount of 200mg, and b. S- Acetyl Glutathione in an amount of 300mg.
  • the present invention relates to an oral pharmaceutical composition comprising a. L-Carnosine in an amount of 200mg, and b. S- Acetyl Glutathione in an amount of lOOmg.
  • the present invention relates to a method of treating disease or disorder in patient in need of such therapy, wherein treatment comprises administration of a. therapeutically effective amount of L-Carnosine or pharmaceutically acceptable salts or derivatives thereof, and b. therapeutically effective amount of Glutathione, its precursors or pharmaceutically acceptable salts or derivatives thereof, in same or different compositions to humans or animal.
  • the present invention relates to a method of treating disease or disorder in patient in need of such therapy, wherein treatment comprises administration of a. therapeutically effective amount of L-Carnosine or pharmaceutically acceptable salts or derivatives thereof, and b. therapeutically effective amount of S-Acetyl Glutathione or pharmaceutically acceptable salts or derivatives thereof, in same or different compositions to humans or animal.
  • the present invention relates to a method of treating Diabetic Nephropathy or Chronic Kidney Disease, wherein treatment comprises administration of a. therapeutically effective amount of L-Carnosine or pharmaceutically acceptable salts or derivatives thereof, and b. therapeutically effective amount of S-Acetyl Glutathione or pharmaceutically acceptable salts or derivatives thereof, in same or different compositions to humans or animal.
  • the present invention relates to a method of treating Diabetic Nephropathy or Chronic Kidney Disease, wherein treatment comprises administration of an oral pharmaceutical composition comprising a. L-Carnosine in an amount between 50mg to 400mg, and b. S- Acetyl Glutathione in an amount between lOOmg to 600mg,
  • a pharmaceutical composition as per present invention includes immediate release, delayed release, extended release or combination thereof.
  • a pharmaceutical composition as per present invention includes for oral, intravenous or other routes of administration.
  • a pharmaceutical composition as per present invention includes solid, liquid, semisolid or other dosage forms.
  • Carnosine is an endogenous dipeptide, composed of b-alanine and L- histidine. Carnosine has pH-buffering, metal-ion chelation, and antioxidant capacity as well as the capacity to protect against formation of advanced glycation and lipoxidation end-products. Carnosine is found naturally mainly in the skeletal muscles, central nervous system, olfactory neurons and in the lens of the eye in some vertebrates, including humans.
  • CNDP1 gene is responsible for formation of camosinase into the serum.
  • Carnosinase is responsible for formation of AGES (Advanced Glycation end products).
  • the AGEs blocks the nephrons during the condition of diabetes.
  • Low carnosinase secretion has been associated with low risk of developing diabetic nephropathy.
  • Low levels of carnosinase secretion means less of this enzyme in the blood, which results in higher concentration of carnosine. It is known that carnosine prevents the formation of Advanced Glycation End products (AGEs) which cause kidney damage in diabetics.
  • AGEs Advanced Glycation End products
  • L-Carnosine or pharmaceutically acceptable salts or derivatives thereof is in the equivalent therapeutically effective amount of L-Carnosine. In a preferred embodiment, L-Carnosine or pharmaceutically acceptable salts or derivatives thereof is in an amount equivalent to 50mg to 400mg of L-Camosine.
  • Glutathione g-L-glutamyl-L-cysteinylglycine, GSH
  • GSH glutathione
  • GSH Reduced glutathione
  • Glutathione derivatives are modified glutathione which may act as a prodrug or have similar pharmacological properties as glutathione.
  • S-Acetyl-glutathione is a GSH analog which is more stable in plasma, it is taken up directly by the cells and later converted to GSH.
  • S-Acetyl Glutathione or pharmaceutically acceptable salts or derivatives thereof is administered in the equivalent therapeutically effective amount of glutathione.
  • S-Acetyl Glutathione or pharmaceutically acceptable salts or derivatives thereof is in an amount equivalent to lOOmg to 600mg of S-Acetyl Glutathione.
  • Glutathione presursors are compound involved in biochemical formation of glutathione in vivo.
  • Such precursors includes but are not limited to glutamic acid or glycine or cysteine or derivatives thereof.
  • Three essential amino acids glutamate, cysteine, and glycine combine to form the tripeptide glutathione.
  • cysteine is joined to glutamate through the action of glutamate cysteine ligase to produce g-glutamylcysteine, which proceeds to link with glycine via glutathione synthase action.
  • N-Acetylcysteine is an exogenous antioxidant, which works as a free radical scavenger. It is also a glutathione precursor regarded as one of the most important intracellular antioxidants. It has been proven that supplementation with N- acetylcysteine activates antioxidant enzymes, prevents oxidative stress development, improves sensitivity towards insulin.
  • the pharmaceutical compositions are the different type of medicinal preparation designed for the administration of targeted one or more drugs.
  • the pharmaceutical compositions as per present invention includes immediate release, delayed release, extended release and pulsed-release.
  • the pharmaceutical compositions can be prepared using uniform mixture of two or more drugs.
  • pharmaceutical composition can be prepared with one or more drugs in separate compartment within a single dosage form.
  • the pharmaceutical compositions as per present invention can be administered by oral, intravenous or other routes of drug administration.
  • the pharmaceutical compositions as per present invention can be solid, liquid, semisolid or any other dosage form.
  • the pharmaceutical compositions as per present invention can be prepared as one or more drug in modified release and other drugs as immediate release in single dosage form.
  • the oral pharmaceutical dosage form are tablets, capsules, solutions, emulsions, suspensions, syrups, elixirs, powders and granules for reconstitution, lozenges, dispersible powders and granules, medicated gums, chewing tablets, effervescent tablets, multi-particulate dosage forms and the likes.
  • the multicompartment dosage form are bilayer tablets, capsule-in-capsule, tablet-in capsule and any other dosage form.
  • the pharmaceutical compositions can be formulated by any techniques known to or appreciated by a person skilled in the art
  • the oral pharmaceutical composition further includes optionally any one or a combination of one or more pharmaceutically acceptable excipients, such as but not limited to carriers, diluents, fillers, disintegrants, lubricating agents, binders, colorants, pigments, stabilizers, preservatives, antioxidants and solubility enhancers.
  • pharmaceutically acceptable excipients such as but not limited to carriers, diluents, fillers, disintegrants, lubricating agents, binders, colorants, pigments, stabilizers, preservatives, antioxidants and solubility enhancers.
  • Example 1 - H202 scavenging assay A solution of hydrogen peroxide was prepared in phosphate buffer. The concentration of hydrogen peroxide was determined by absorption at 230 nm using a spectrophotometer. Extract in distilled water was added to hydrogen peroxide and absorbance at 230 nm is determined after 10 min against a blank solution containing phosphate buffer without hydrogen peroxide. The ascorbic acid was used as standard in the assay. The test and standard solutions were prepared in millipore water. Different concentrations of the test and standard solutions were used in the assay.
  • the IC50 value of ascorbic acid was found to be 312.29 ⁇ 11.79 mM for 20 mM H202 solution.
  • the IC50 value of L-carnosine was found to be 1332.83 ⁇ 84.44 mM for 20 mM H202 solution.
  • the IC50 value of S-acetyl 1-glutathione was found to be 309.95 ⁇ 6.27 mM for 20 mM H202 solution.
  • the change in optical density of DPPH radicals was monitored.
  • the sample was diluted with methanol and DPPH solution was added. After 30 min, the absorbance is measured at 517 nm.
  • the ascorbic acid was used as standard in the assay.
  • the test and standard solutions were prepared in millipore water. Different concentrations of the test and standard solutions were used in the assay.
  • the DPPH solution (100 mM) was prepared in methanol.
  • the % inhibition of DPPH was calculated using following formula.
  • the IC50 value of ascorbic acid was found to be 7.31 ⁇ 0.068 pg/ml for 100 mM DPPH solution.
  • the IC50 value of L-carnosine was found to be 949.37 ⁇ 74.78 pg/ml for 100 mM DPPH solution.
  • the IC50 value of S-acetyl 1-glutathione was found to be 13.0 ⁇ 0.44 pg/ml for 100 mM DPPH solution.
  • kidney homogenate was used for estimation of renal oxidative stress parameters.
  • Direct kidney homogenate was used for estimation of malonaldehyde (MDA) and reduced glutathione (GSH).
  • MDA malonaldehyde
  • GSH reduced glutathione
  • Post nuclear supernatant was used for determination of catalase activity while post mitochondrial supernatant was used for determination of superoxide dismutase (SOD).
  • SOD superoxide dismutase
  • the lipid peroxide level in animal tissues was measured according to method described by Ohkawa and co-workers (Ohkawa et ah, 1979).
  • the diabetic control group showed significant increase in MDA levels when compared to normal control group.
  • the treatment with 1-carnosine for 28 days at a dose of 36 mg/kg showed significant decrease in MDA when compared to diabetic control group.
  • the animals treated with s-acetyl-l-glutathione at all selected dose levels showed significant decrease in MDA when compared with diabetic control group.
  • Combination group also showed significant decrease in MDA when compared to diabetic control animals (Figure 3).
  • Catalase activity was assayed according to the method of Luck (Luck, 1965).
  • Enzyme activity was measured using molar extinction coefficient of H2O2 (0.071) and was expressed as micromole of H2O2 decomposed/min/ mg protein)
  • the diabetic control group showed significant decrease in catalase levels when compared to normal control group.
  • the animals treated with s-acetyl-l-glutathione at all selected dose levels showed significant increase in catalase levels when compared with diabetic control group.
  • Combination group also showed significant increase in catalase when compared to diabetic control animals (Figure 4).
  • One unit of enzyme activity is defined as the concentration of enzyme that gives half maximal inhibition of nitrobluetetrazolium (NBT) reduction. Result was expressed as U/ mg protein.
  • control and treatment groups were analysed using graph pad prism V5.0 for Windows.
  • significance level between control and experimental groups was assessed by one-way analysis of variance (ANOVA) followed by Dunnett’s multiple comparison test. All data were expressed as Mean ⁇ Standard Error of Mean (SEM); the P values less than 0.05 were considered to be significant.
  • the diabetic control group showed significant decrease in superoxide dismutase (SOD) levels when compared to normal control group.
  • SOD superoxide dismutase
  • the treatment with 1- carnosine for 28 days at a dose of 36 mg/kg showed significant increase in SOD levels when compared to diabetic control group.
  • the animals treated with s-acetyl- 1-glutathione at all selected dose levels showed significant increase in SOD levels when compared with diabetic control group.
  • Combination group also showed significant increase in SOD when compared to diabetic control animals (Figure 5).
  • Example - 4 Representative tablet composition of L- Camosine and S-Acetyl Glutathione .

Landscapes

  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Biochemistry (AREA)
  • Veterinary Medicine (AREA)
  • Molecular Biology (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Genetics & Genomics (AREA)
  • Public Health (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Biophysics (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Immunology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

The present invention relates to pharmaceutical composition comprising L- Carnosine or pharmaceutically acceptable salts or derivatives thereof, and Glutathione, its precursors or pharmaceutically acceptable salts or derivatives thereof. The present invention also relates to method of treating disease or disorder in patient in need of such therapy, wherein treatment comprises administration of L-Carnosine or pharmaceutically acceptable salts or derivatives thereof, and Glutathione, its precursors or pharmaceutically acceptable salts or derivatives thereof, in same or different compositions to humans or animal. The pharmaceutical composition is helpful particularly in oral administrations. It is useful is the treatment of Diabetic Nephropathy and Chronic Kidney Disease.

Description

A PHARMACEUTICAL COMPOSITION AND METHOD OF TREATMENT USING L-CARNOSINE AND GLUTATHIONE, ITS PRECURSORS OR DERIVATIVES THEREOF.
The present invention relates to pharmaceutical composition comprising L- Carnosine or pharmaceutically acceptable salts or derivatives thereof, and Glutathione, its precursors or pharmaceutically acceptable salts or derivatives thereof. The present invention also relates to method of treating disease or disorder in patient in need of such therapy, wherein treatment comprises administration of L-Carnosine or pharmaceutically acceptable salts or derivatives thereof, and Glutathione, its precursors or pharmaceutically acceptable salts or derivatives thereof, in same or different compositions to humans or animal.
BACKGROUND OF INVENTION
Oxidative stress, ion balance and pH balance are some the most critical factors involved in the major acute and chronic physiological imbalance. The depletion of powerful antioxidant renders cells particularly vulnerable to oxidative stress. The resulting damage is the key step in the onset and progression of many disease states.
Several endogenous and exogenous compound are known to protect against oxidative stress. These endogenous and exogenous compound are combined with other therapies like antibiotics, anti-cancer drugs to achieved combined desired therapy goals.
L-Carnosine is used as protective agent for many therapies. Glutathione is endogenous agent involved in anti-aging and other biochemical pathways providing shielding effect upon administration. While both these compounds are known to be administrated individual for their respective mechanism of action, individual administration has a limitation of affecting only their respective physiological involvement.
Therefore, there is an urgent need for the improved therapy with high anti oxidant, anti-aging protective effect for many disease or disorders including diabetic nephropathy and chronic kidney disease.
Till date, pharmaceutical composition comprising L-Carnosine or pharmaceutically acceptable salts or derivatives thereof, and Glutathione, its precursors or pharmaceutically acceptable salts or derivatives thereof are not known.
DESCRIPTION OF THE DRAWINGS
Figure 1 : Effect of combination of L- camosine and S-acetyl-L-glutathione on H2O2 scavenging activity.
Figure : 2 Effect of combination of L- carnosine and S-acetyl-L-glutathione on DPPH Scavenging activity.
Figure 3: Effect of 1-camosine and s-acetyl -1-glutathione on MDA levels in kidney tissues.
Figure 4: Effect of 1-camosine and s-acetyl-l-glutathione on catalase levels in kidney tissue.
Figure 5: Effect of 1-camosine and s-acetyl-l-glutathione on SOD levels in kidney tissues.
DETAILED DESCRIPTION OF THE INVENTION
The present invention relates to pharmaceutical composition comprising L- Carnosine or pharmaceutically acceptable salts or derivatives thereof, and Glutathione, its precursors or pharmaceutically acceptable salts or derivatives thereof. The active ingredient as per present invention are used in therapeutically effective amount.
"Therapeutically effective amount" or "effective amount" refers to the amount of a pharmaceutically active agent when administered to a patient, is sufficient to affect such treatment for the disease. The therapeutically effective amount will vary depending on the disease and its severity, and the age, weight, and other conditions of the patient to be treated.
The term "pharmaceutical compositions" herein refers to any composition for administration to human or animal by any route and includes but are not limited to immediate release, delayed release, extended release and pulsed-release.
In an embodiment, the present invention relates to a pharmaceutical composition comprising a. therapeutically effective amount of L-Carnosine or pharmaceutically acceptable salts or derivatives thereof, and b. therapeutically effective amount of Glutathione, its precursors or pharmaceutically acceptable salts or derivatives thereof.
In an embodiment, the present invention relates to a pharmaceutical composition consisting of a. therapeutically effective amount of L-Carnosine or pharmaceutically acceptable salts or derivatives thereof, and b. therapeutically effective amount of Glutathione, its precursors or pharmaceutically acceptable salts or derivatives thereof c. one or more suitable inactive ingredients.
In an embodiment, the present invention relates to a pharmaceutical composition comprising c. therapeutically effective amount of L-Carnosine or pharmaceutically acceptable salts or derivatives thereof, and d. therapeutically effective amount of S-Acetyl Glutathione or pharmaceutically acceptable salts or derivatives thereof.
In an embodiment, the present invention relates to a pharmaceutical composition consisting of d. therapeutically effective amount of L-Carnosine or pharmaceutically acceptable salts or derivatives thereof, and e. therapeutically effective amount of S-Acetyl Glutathione or pharmaceutically acceptable salts or derivatives thereof f. one or more suitable inactive ingredients.
In a preferred embodiment, the present invention relates to a pharmaceutical comprising a. L-Carnosine or pharmaceutically acceptable salts or derivatives thereof in an amount equivalent to 50mg to 400mg of L-Carnosine, and b. S-Acetyl Glutathione or pharmaceutically acceptable salts or derivatives thereof in an amount equivalent to lOOmg to 600mg of
S-Acetyl Glutathione.
In a preferred embodiment, the present invention relates to an oral pharmaceutical composition comprising a. L-Carnosine in an amount between 50mg to 400mg, and b. S- Acetyl Glutathione in an amount between lOOmg to 600mg.
In a preferred embodiment, the present invention relates to an oral pharmaceutical composition comprising a. L-Carnosine in an amount of 200mg, and b. S- Acetyl Glutathione in an amount of 300mg. In a preferred embodiment, the present invention relates to an oral pharmaceutical composition comprising a. L-Carnosine in an amount of 200mg, and b. S- Acetyl Glutathione in an amount of lOOmg.
In an embodiment, the present invention relates to a method of treating disease or disorder in patient in need of such therapy, wherein treatment comprises administration of a. therapeutically effective amount of L-Carnosine or pharmaceutically acceptable salts or derivatives thereof, and b. therapeutically effective amount of Glutathione, its precursors or pharmaceutically acceptable salts or derivatives thereof, in same or different compositions to humans or animal.
In an embodiment, the present invention relates to a method of treating disease or disorder in patient in need of such therapy, wherein treatment comprises administration of a. therapeutically effective amount of L-Carnosine or pharmaceutically acceptable salts or derivatives thereof, and b. therapeutically effective amount of S-Acetyl Glutathione or pharmaceutically acceptable salts or derivatives thereof, in same or different compositions to humans or animal.
In an embodiment, the present invention relates to a method of treating Diabetic Nephropathy or Chronic Kidney Disease, wherein treatment comprises administration of a. therapeutically effective amount of L-Carnosine or pharmaceutically acceptable salts or derivatives thereof, and b. therapeutically effective amount of S-Acetyl Glutathione or pharmaceutically acceptable salts or derivatives thereof, in same or different compositions to humans or animal.
In an embodiment, the present invention relates to a method of treating Diabetic Nephropathy or Chronic Kidney Disease, wherein treatment comprises administration of an oral pharmaceutical composition comprising a. L-Carnosine in an amount between 50mg to 400mg, and b. S- Acetyl Glutathione in an amount between lOOmg to 600mg,
In one or more embodiments, a pharmaceutical composition as per present invention includes immediate release, delayed release, extended release or combination thereof.
In one or more embodiments, a pharmaceutical composition as per present invention includes for oral, intravenous or other routes of administration.
In one or more embodiments, a pharmaceutical composition as per present invention includes solid, liquid, semisolid or other dosage forms. L- CARNOSINE
Carnosine is an endogenous dipeptide, composed of b-alanine and L- histidine. Carnosine has pH-buffering, metal-ion chelation, and antioxidant capacity as well as the capacity to protect against formation of advanced glycation and lipoxidation end-products. Carnosine is found naturally mainly in the skeletal muscles, central nervous system, olfactory neurons and in the lens of the eye in some vertebrates, including humans.
CNDP1 gene is responsible for formation of camosinase into the serum. Carnosinase is responsible for formation of AGES (Advanced Glycation end products). The AGEs blocks the nephrons during the condition of diabetes. Low carnosinase secretion has been associated with low risk of developing diabetic nephropathy. Low levels of carnosinase secretion means less of this enzyme in the blood, which results in higher concentration of carnosine. It is known that carnosine prevents the formation of Advanced Glycation End products (AGEs) which cause kidney damage in diabetics.
In one or more embodiments, L-Carnosine or pharmaceutically acceptable salts or derivatives thereof is in the equivalent therapeutically effective amount of L-Carnosine. In a preferred embodiment, L-Carnosine or pharmaceutically acceptable salts or derivatives thereof is in an amount equivalent to 50mg to 400mg of L-Camosine.
GLUTATHIONE OR ITS PRECURSORS OR DERIVATIVES
Glutathione (g-L-glutamyl-L-cysteinylglycine, GSH), a water-soluble endogenous tripeptide, is the most abundant thiol present in mammalian cells. The liver has highest content of this tripeptide and is the main tissue participating in GSH biosynthesis. Glutathione helps remove the toxins and free radical scavenges from the body by the oxidation process. The kidneys are highly dependent on an adequate supply of glutathione (GSH) to maintain normal function. This is due, in part, to high rates of aerobic metabolism, particularly in the proximal tubules.
Reduced glutathione (GSH) is the preferred substrate for several enzymes in xenobiotic metabolism and antioxidant defence. It plays an important role in many cellular processes, such as cell differentiation, proliferation and apoptosis. GSH deficiency has been observed in aging and in a wide range of pathologies.
Glutathione derivatives are modified glutathione which may act as a prodrug or have similar pharmacological properties as glutathione. S-Acetyl-glutathione is a GSH analog which is more stable in plasma, it is taken up directly by the cells and later converted to GSH. In one or more embodiments, S-Acetyl Glutathione or pharmaceutically acceptable salts or derivatives thereof is administered in the equivalent therapeutically effective amount of glutathione. In a preferred embodiment, S-Acetyl Glutathione or pharmaceutically acceptable salts or derivatives thereof is in an amount equivalent to lOOmg to 600mg of S-Acetyl Glutathione.
Glutathione presursors are compound involved in biochemical formation of glutathione in vivo. Such precursors includes but are not limited to glutamic acid or glycine or cysteine or derivatives thereof. Three essential amino acids glutamate, cysteine, and glycine combine to form the tripeptide glutathione. At the beginning, cysteine is joined to glutamate through the action of glutamate cysteine ligase to produce g-glutamylcysteine, which proceeds to link with glycine via glutathione synthase action.
N-Acetylcysteine is an exogenous antioxidant, which works as a free radical scavenger. It is also a glutathione precursor regarded as one of the most important intracellular antioxidants. It has been proven that supplementation with N- acetylcysteine activates antioxidant enzymes, prevents oxidative stress development, improves sensitivity towards insulin.
PHARMACEUTICAL COMPOSITION
The pharmaceutical compositions are the different type of medicinal preparation designed for the administration of targeted one or more drugs. The pharmaceutical compositions as per present invention includes immediate release, delayed release, extended release and pulsed-release. The pharmaceutical compositions can be prepared using uniform mixture of two or more drugs. In one or more embodiments, pharmaceutical composition can be prepared with one or more drugs in separate compartment within a single dosage form. The pharmaceutical compositions as per present invention can be administered by oral, intravenous or other routes of drug administration. The pharmaceutical compositions as per present invention can be solid, liquid, semisolid or any other dosage form. The pharmaceutical compositions as per present invention can be prepared as one or more drug in modified release and other drugs as immediate release in single dosage form.
The oral pharmaceutical dosage form are tablets, capsules, solutions, emulsions, suspensions, syrups, elixirs, powders and granules for reconstitution, lozenges, dispersible powders and granules, medicated gums, chewing tablets, effervescent tablets, multi-particulate dosage forms and the likes. The multicompartment dosage form are bilayer tablets, capsule-in-capsule, tablet-in capsule and any other dosage form. The pharmaceutical compositions can be formulated by any techniques known to or appreciated by a person skilled in the art
In an embodiment, the oral pharmaceutical composition further includes optionally any one or a combination of one or more pharmaceutically acceptable excipients, such as but not limited to carriers, diluents, fillers, disintegrants, lubricating agents, binders, colorants, pigments, stabilizers, preservatives, antioxidants and solubility enhancers.
Having described the invention with reference to the different embodiments of the invention, other embodiments will become apparent to one skilled in the art from consideration of the specifications.
The innovation is further defined by reference to the following examples. It will be apparent to those skilled in the art that many modifications, both to the composition and treatment, may be practiced without departing from the scope of this invention. EXAMPLES
Example 1 - H202 scavenging assay. A solution of hydrogen peroxide was prepared in phosphate buffer. The concentration of hydrogen peroxide was determined by absorption at 230 nm using a spectrophotometer. Extract in distilled water was added to hydrogen peroxide and absorbance at 230 nm is determined after 10 min against a blank solution containing phosphate buffer without hydrogen peroxide. The ascorbic acid was used as standard in the assay. The test and standard solutions were prepared in millipore water. Different concentrations of the test and standard solutions were used in the assay.
The % scavenging of H2O2 was calculated using following formula. JC _ % Scavenging of H202 = — - - X100 iC where Ac is absorbance of Control; At is absorbance of test after reduction of blank reading from test reading.
Preparation of standard and test solution for H2O2 assay
Figure imgf000011_0001
The 40 mM solution was prepared in 50 mM phosphate buffer (pH - 7.4) Assay Procedure:
Figure imgf000012_0001
Results
The IC50 value of ascorbic acid was found to be 312.29±11.79 mM for 20 mM H202 solution.
The IC50 value of L-carnosine was found to be 1332.83±84.44 mM for 20 mM H202 solution. The IC50 value of S-acetyl 1-glutathione was found to be 309.95±6.27 mM for 20 mM H202 solution.
Effect of combination of L- carnosine and S-acetyl-L-glutathione on H2O2 scavenging activity
Figure imgf000012_0002
Figure imgf000013_0002
Data are shown as Mean± SEM (n=3).
Example 2 - DPPH Assay.
In order to evaluate the antioxidant potential through free radical scavenging by the test samples, the change in optical density of DPPH radicals was monitored. The sample was diluted with methanol and DPPH solution was added. After 30 min, the absorbance is measured at 517 nm. The ascorbic acid was used as standard in the assay. The test and standard solutions were prepared in millipore water. Different concentrations of the test and standard solutions were used in the assay. The DPPH solution (100 mM) was prepared in methanol.
The % inhibition of DPPH was calculated using following formula.
Figure imgf000013_0001
Where Ac is absorbance of Control; At is absorbance of test. Preparation of standard and test solution for DPPH assay
Figure imgf000014_0001
Assay Procedure:
Figure imgf000014_0002
Results
The IC50 value of ascorbic acid was found to be 7.31±0.068 pg/ml for 100 mM DPPH solution. The IC50 value of L-carnosine was found to be 949.37±74.78 pg/ml for 100 mM DPPH solution.
The IC50 value of S-acetyl 1-glutathione was found to be 13.0±0.44 pg/ml for 100 mM DPPH solution.
Effect of combination of L- carnosine and S-acetyl-L-glutathione on DPPH Scavenging activity
Figure imgf000015_0001
Data are shown as Mean± SEM (n=3). Example - 3 In-vivo Studies - Assessment of renal oxidative stress parameters.
The kidney homogenate was used for estimation of renal oxidative stress parameters. Direct kidney homogenate was used for estimation of malonaldehyde (MDA) and reduced glutathione (GSH). Post nuclear supernatant was used for determination of catalase activity while post mitochondrial supernatant was used for determination of superoxide dismutase (SOD). MALONDI ALDEHYDE (MDA) ASSAY
The lipid peroxide level in animal tissues was measured according to method described by Ohkawa and co-workers (Ohkawa et ah, 1979).
Procedure
Preparation of assay standard and solutions
Stock was prepared by adding 17 mΐ of TMP (1,1,3,3-tetramethoxy propane) to 983 mΐ of distilled water to give 100 mM solution of TMP. Serial dilutions were done by diluting the stock with the distilled water to get 1, 3, 10, 30, 100 and 300 mM solutions of TMP. The 2-thiobarbituric acid solution was prepared by dissolving required quantity in 0.1 M phosphate buffer (pH- 7.4). Sodium dodecyl sulphate solution and glacial acetic acid solution was prepared in distilled water. Assay procedure of MDA assay
Figure imgf000016_0001
Calibration curve of standard was plotted and unknown concentrations of the MDA in samples were calculated. The results were expressed as amount of malondialdehyde formed as nmoles of MDA/ mg protein.
Results The diabetic control group showed significant increase in MDA levels when compared to normal control group. The treatment with 1-carnosine for 28 days at a dose of 36 mg/kg showed significant decrease in MDA when compared to diabetic control group. The animals treated with s-acetyl-l-glutathione at all selected dose levels showed significant decrease in MDA when compared with diabetic control group. Combination group also showed significant decrease in MDA when compared to diabetic control animals (Figure 3).
CATALASE ASSAY
Determination of catalase activity was carried by using post nuclear supernatant. Catalase activity was assayed according to the method of Luck (Luck, 1965).
Procedure Table 1 Determination of catalase activity
Figure imgf000017_0001
Calculations
Enzyme activity was measured using molar extinction coefficient of H2O2 (0.071) and was expressed as micromole of H2O2 decomposed/min/ mg protein)
Results
The diabetic control group showed significant decrease in catalase levels when compared to normal control group. The animals treated with s-acetyl-l-glutathione at all selected dose levels showed significant increase in catalase levels when compared with diabetic control group. Combination group also showed significant increase in catalase when compared to diabetic control animals (Figure 4).
SUPEROXIDE DISMUTASE (SOD)
Estimation of SOD was carried out by using post mitochondrial supernatant (PMS). SOD was estimated according to method described by Paoletti and Mocali (Paoletti et al., 1986). Procedure
Estimation of Superoxide dismutase
Figure imgf000018_0001
Calculation
One unit of enzyme activity is defined as the concentration of enzyme that gives half maximal inhibition of nitrobluetetrazolium (NBT) reduction. Result was expressed as U/ mg protein.
Statistical analysis
The differences among control and treatment groups were analysed using graph pad prism V5.0 for Windows. The significance level between control and experimental groups was assessed by one-way analysis of variance (ANOVA) followed by Dunnett’s multiple comparison test. All data were expressed as Mean ± Standard Error of Mean (SEM); the P values less than 0.05 were considered to be significant.
Results
The diabetic control group showed significant decrease in superoxide dismutase (SOD) levels when compared to normal control group. The treatment with 1- carnosine for 28 days at a dose of 36 mg/kg showed significant increase in SOD levels when compared to diabetic control group. The animals treated with s-acetyl- 1-glutathione at all selected dose levels showed significant increase in SOD levels when compared with diabetic control group. Combination group also showed significant increase in SOD when compared to diabetic control animals (Figure 5). Example - 4 Representative tablet composition of L- Camosine and S-Acetyl Glutathione .
Figure imgf000019_0001

Claims

CLAIMS aim
1. A pharmaceutical composition comprising a. L-Carnosine or pharmaceutically acceptable salts or derivatives thereof, and b. Glutathione, its precursors or pharmaceutically acceptable salts or derivatives thereof.
2. The pharmaceutical composition as claimed in claim 1, wherein said glutathione derivative is S-Acetyl Glutathione or pharmaceutically acceptable salts or derivatives thereof
3. The pharmaceutical composition as claimed in claim 1, wherein said Glutathione derivative is S-Acetyl Glutathione or pharmaceutically acceptable salts or derivatives thereof, and wherein said L-Carnosine or pharmaceutically acceptable salts or derivatives thereof is in an amount equivalent to 50mg to 400mg of L- Carnosine, and S-Acetyl Glutathione or pharmaceutically acceptable salts or derivatives thereof is in an amount equivalent to lOOmg to 600mg of S- Acetyl Glutathione.
4. The pharmaceutical composition as claimed in claim 1, wherein said pharmaceutical composition is an oral pharmaceutical composition comprising L-Camosine in an amount between 50mg to 400mg, and S- Acetyl Glutathione in an amount between lOOmg to 600mg.
5. The pharmaceutical composition as claimed in claim 4, wherein said L- Carnosine in an amount of 200mg, and S-Acetyl Glutathione in an amount of lOOmg.
6. The pharmaceutical composition as claimed in claim 1, wherein at least one of (a) and (b) is in delayed release or extended-release form.
7. The pharmaceutical composition as claimed in claim 1, wherein (a) and (b) are in separate compartment within a single dosage form.
8. The pharmaceutical composition as claimed in claim 1, wherein said pharmaceutical composition is tablets.
9. A pharmaceutical composition comprising a. therapeutically effective amount of L-Carnosine or pharmaceutically acceptable salts or derivatives thereof, and b. therapeutically effective amount of Glutathione, its precursors or pharmaceutically acceptable salts or derivatives thereof.
10. A method of treating disease or disorder in patient in need of such therapy, wherein treatment comprises administration of a. L-Carnosine or pharmaceutically acceptable salts or derivatives thereof, and b. Glutathione, its precursors or pharmaceutically acceptable salts or derivatives thereof, in same or different compositions to humans or animal.
11. The method of treating as claimed in claim 10, wherein said treatment comprises administration of S-Acetyl Glutathione or pharmaceutically acceptable salts or derivatives thereof.
12. The method of treating as claimed in claim 10, wherein said treatment comprises administration of L-Carnosine or pharmaceutically acceptable salts or derivatives thereof is in an amount equivalent to 50mg to 400mg of L-Carnosine, and S-Acetyl Glutathione or pharmaceutically acceptable salts or derivatives thereof is in an amount equivalent to lOOmg to 600mg of S- Acetyl Glutathione.
13. The method of treating as claimed in claim 10, wherein said treatment is for treating Diabetic Nephropathy or Chronic Kidney Disease.
PCT/IB2021/053397 2020-04-26 2021-04-25 A pharmaceutical composition and method of treatment using l-carnosine and glutathione, its precursors or derivatives thereof WO2021220124A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
IN202021017799 2020-04-26
IN202021017799 2020-04-26

Publications (1)

Publication Number Publication Date
WO2021220124A1 true WO2021220124A1 (en) 2021-11-04

Family

ID=78332323

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/IB2021/053397 WO2021220124A1 (en) 2020-04-26 2021-04-25 A pharmaceutical composition and method of treatment using l-carnosine and glutathione, its precursors or derivatives thereof

Country Status (1)

Country Link
WO (1) WO2021220124A1 (en)

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20170312329A1 (en) * 2016-04-25 2017-11-02 William H. Cross, III Compositions and Methods for Treatment of Prediabetes
CN107823095A (en) * 2017-12-07 2018-03-23 安徽生物肽产业研究院有限公司 A kind of active peptide facial mask
CN108078804A (en) * 2017-12-20 2018-05-29 广东圣蜜生物科技有限公司 A kind of composite humectant and preparation method thereof
CN108635273A (en) * 2018-07-12 2018-10-12 太原紫兰科技有限责任公司 A kind of sun-proof sun screen of wrinkle resistance, whitening and preparation method thereof

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20170312329A1 (en) * 2016-04-25 2017-11-02 William H. Cross, III Compositions and Methods for Treatment of Prediabetes
CN107823095A (en) * 2017-12-07 2018-03-23 安徽生物肽产业研究院有限公司 A kind of active peptide facial mask
CN108078804A (en) * 2017-12-20 2018-05-29 广东圣蜜生物科技有限公司 A kind of composite humectant and preparation method thereof
CN108635273A (en) * 2018-07-12 2018-10-12 太原紫兰科技有限责任公司 A kind of sun-proof sun screen of wrinkle resistance, whitening and preparation method thereof

Similar Documents

Publication Publication Date Title
JP5736421B2 (en) Regulation of physiological processes and drugs useful for this
Tsay et al. Age-associated changes of superoxide dismutase and catalase activities in the rat brain
US9427419B2 (en) Compositions comprising dimethyl sulfoxide (DMSO)
Zavodnik et al. Melatonin and succinate reduce rat liver mitochondrial dysfunction in diabetes
US20190151233A1 (en) Dopa decarboxylase inhibitor compositions
EP3124023B1 (en) Durable preparation of an injectable of melatonin exhibiting long-term stability
WO2009014624A2 (en) Nrf2 inducers for treating epidermolysis bullosa simplex
US20210000777A1 (en) Dopa decarboxylase inhibitor compositions
PT1708711E (en) Compositions useful especially for treatment or prevention of metabolic syndrome
US20090312238A1 (en) Protection Against Oxidative Damage in Cells
TW201605434A (en) Use of cysteamine and derivatives thereof to treat mitochondrial diseases
US6451761B1 (en) N′N′-dichlorinated omega-amino acids and uses thereof
WO2021220124A1 (en) A pharmaceutical composition and method of treatment using l-carnosine and glutathione, its precursors or derivatives thereof
Mayayo-Vallverdú et al. The antioxidant l-Ergothioneine prevents cystine lithiasis in the Slc7a9−/− mouse model of cystinuria
AU2022339876A1 (en) Methods, systems and compositions for inhibition of cellular dysfunction and cell death with deuterated pufas
US8664237B2 (en) Spiperone derivatives and methods of treating disorders
KR100929979B1 (en) Novel herbal composition with anticancer side effects and anticancer activity
US20110003891A1 (en) Novel use of zinc n-acetyltaurinate
US20210085676A1 (en) Treatment for age- and oxidative stress-associated muscle atrophy and weakness
TWI672145B (en) Use and pharmaceutical composition for metabolic disease prevention and/or treatment
US20110118327A1 (en) In vivo assessment of toxicity and pharmacokinetics of methylglyoxal
Helal et al. Antioxidants Reveal Protection against the Biochemical Changes in Liver, Kidney and Blood Profiles after Clindamycin/Ibuprofen Administration in Dental Patients
WO2018139477A1 (en) NOVEL THERAPEUTIC AGENT FOR NEPHROGENIC ANEMIA TARGETING ERYTHROPOIETIN RECEPTOR VIA NO AND NO-pathway STIMULATION
US20050009914A1 (en) Use of a cysteine-containing substance to increase the ventilatory activity and erythropoientin production
Asagba A comparative study on the biochemical effect of ocular and oral cadmium administration in rabbits

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 21796944

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 21796944

Country of ref document: EP

Kind code of ref document: A1

32PN Ep: public notification in the ep bulletin as address of the adressee cannot be established

Free format text: NOTING OF LOSS OF RIGHTS PURSUANT TO RULE 112(1) EPC (EPO FORM 1205A DATED 07/07/2023)

122 Ep: pct application non-entry in european phase

Ref document number: 21796944

Country of ref document: EP

Kind code of ref document: A1