Title of Invention	A PROCESS OF PREPARING A NEW ANTI-OXIDANT COMPOSITION FOR THE TREATMENT OF CHRONIC RENAL FAILURE AND HEART FAILURE
Abstract	This invention provides a process of preparing a new antioxidant composition for the treatment of chronic renal failure and also for heart failure comprising mixing the following anti-oxidants in the proportions indicated there against: Coenzyme Q10 - 10-40% by wt. Vitamin E - 30-45% by wt. Vitamin C - 30-45% by wt. and the balance if any adding a conventional filler for preparing the tablet or capsule.

Title of Invention

A PROCESS OF PREPARING A NEW ANTI-OXIDANT COMPOSITION FOR THE TREATMENT OF CHRONIC RENAL FAILURE AND HEART FAILURE

Abstract

This invention provides a process of preparing a new antioxidant composition for the treatment of chronic renal failure and also for heart failure comprising mixing the following anti-oxidants in the proportions indicated there against: Coenzyme Q10 - 10-40% by wt. Vitamin E - 30-45% by wt. Vitamin C - 30-45% by wt. and the balance if any adding a conventional filler for preparing the tablet or capsule.

Full Text	FORM-3 A THE PATENTS ACT, 1970 i COMPLETE SPECIFICATION SECTION 10 Title:- A PROCESS OF PREPARING A NEW ANTI-OXIDANT COMPOSITION FOR THE TREATMENT OF CHRONIC RENAL FAILURE AND HEART FAILURE, Applicant (8): - PROF, RAM BHADUR SINGH AND DR.MOHAMMAD ARIZ NIAZ, SAFE ENTERPRISES, 6 CITIZEN APARTMENT 30TH ROAD, BANDRA, MUMBAI - 400 050, •MAHARASHTRA, INDIA. The following Specification particularly deadbea and ascertains the nature of this Invention and the manner in which it is to be performed :- This invention relates to a process of preparing a new anti-oxidant composition for the treatment of chronic renal failure and heart failure. BACKGROUND Superoxide and hydroxyl radicals and hydrogen peroxide are important reactive oxygen species in the pathogenesis of renal damage resulting into renal failure.(l-4) Evidence supporting a role for oxidative stress in renal injury has been derived predominantly from studies using a variety of antioxidants such as superoxide dismutase,glutathione peroxidase and catalase.(2-7).Case control studies (8,9) indicate that there is deficiency of antioxidant vitamins A, E and C and beta-carotene as well as coenzyme Q10, lycopene, zinc, selenium in patients with chronic renal failure with and without dialysis as well as in heart failure. However, all studies have not reported a deficiency of antioxidants in renal failure and heart failure. Increased levels of thiobarbituric acid reactive substances (TBARS), and malondialdehyde (MDA) which are indicators of free radical damage have also been described in patients with chronic renal failure and heart failure. However, no study has examined the role of antioxidants in patients with chronic renal failure associated with heart failure. The role of coenzyme Q10 in heart failure is known however the dosages and Efficacy is controversial. The object of this invention is to examine the role of anti-oxidants in patients with chronic renal failure and in a substudy to find out whether it is also beneficial in heart failure and thereafter develop a composition of anti-oxidants for the treatment of chronic renal failure and heart failure. To achieve the said objective, the effects of individual anti-oxidants in moderate, higher, in combination were studied by administering various anti-oxidants in randomized, double blind trials without a control group for a period of four weeks to study the synergistic effects, as given in tables 1-5. A randomized, double blind and controlled trial was conducted in 97 patients to confirm our findings both in renal failure and heart failure (Tables 6-10). Effects of individual antioxidants in moderate doses: We administered coenzyme Q10 (100 mg/day), vitamin E (300 mg/day) and vitamin C(300 mg/day) after randomizing the subjects into three groups (Table 1). There was a modest reduction in serum creatinine and blood urea in the coenzyme Q10 group, doubtful beneficial effect in the vitamin E group and no effect in vitamin C group. The changes were non-significant. Effects of individual antioxidants in higher doses: Subjects were randomized in to three groups in a double blind fashion. They were administered, coenzyme QIO (200 mg/day) or vitamin E (600 mg/day) or vitamin C (600 mg/day) in the concerned group for a period of 4 weeks. Treatment with coenzyme Q10 was associated with substantial reduction in blood urea nitrogen and serum creatinine and increase in creatinine clearance and urine output (Table 2). Treatment with vitamin E also caused substantial changes but slightly lower than coenzyme Q10. Vitamin C had neutral effect. There was significant beneficial effect in patients with heart failure in the coenzume Q10 group with little good effect in vitamin E group and no effect in vitamin C group. Effects of two combined antioxidant formulation in higher doses: Subjects were randomized into three groups (after a wash out period of 4 weeks) in a double blind fashion. They were administered coenzyme QlO+Vitamin E, Coenzyme QlO+Vitamin C and vitamin E + Vitamin C. There was a substantial reduction in blood urea nitrogen and serum creatinine and an increase in creatinine clearance and urine output indicating that some synergistic effect is possible. Patients with heart failure (2 in each group) showed substantial beneficial effect in the first two group and non-significant benefit in the third group. Effect of three combined antioxidant formulations in various doses: Subjects were randomized into three groups as given in table 4 and were administered various combinations of formulations in a double blind fashion for 4 weeks. High dose combination of coenzyme Q10 (200 mg) + vitamin E (600 mg) + vitamin C (600 mg) per day showed a marked reduction in blood urea nitrogen and serum creatinine and an increase in creatinine clearance and urine output as well as in heart function. Moderate dose combination also caused substantial improvement in renal function, and heart function (in patients with heart failure) but lower compared to high dose combination. However lowest dose combination showed only mild nonsignificant benefit in renal function indicating failure of this dosage schedule in reversing renal dysfunction. High and moderate dose groups also caused significant benefit in heart function in patients with heart failure. Finally the results with high dose combination were examined in a randomized, controlled trial in 97 patients of chronic renal failure including 33 with heart failure (Table 6-10).There was a significant benefit in patients with renal failure as well as in patients with heart failure(Table 10).Side effects were mild(Table 9). In view of the above study, a process for preparing a new anti-oxidant composition for the treatment of chronic renal failure and heart failure was developed. According to this invention, the said process of preparing a new anti-oxidant composition for the treatment of chronic renal failure and heart failure comprising mixing the following anti-oxidants in the proportion indicated there against: Coenzyme Q10(Ubiquinone)- 10-40% by wt. Vitamin E - 30-45% by wt. Vitamin C - 30-45% by wt. and the balance if any adding conventional fillers for preparing the tablet or capsule. The filler is used to make the quantity of the composition acceptable and also to fill the capsule. The said filler used is Psvlliul. The proportions of the ingredients used in the preparation of anti-oxidant composition are as follows Coenzyme QIO - 20%bywt. Vitamin E - 40% by wt. Vitamin C - 40%bywt. The proportions of the ingredients in the preparation of anti-oxidant composition are as follows , Coenzyme QIO - 30%bywt. Vitamin E - 30% by wt. Vitamin C - 30% by wt. Filler - 10%bywt. The invention will now be described with reference to the following examples; Example 1: The test agent anti-oxidants, coenzymes QIO, Vitamin C and vitamin E were purchased from the market. These ingredients were mixed in the following proportions and thereafter they were filled in capsules without filler. The capsules prepared were of 250 mg. Coenzyme QIO - 50mg. Vitamin E - lOOmg. Vitamin C - lOOmg Example 2: . The test agent anti-oxidants, coenzymes QIO, Vitamin C and vitamin E were purchased from the market. These ingredients were mixed in the following proportions and thereafter they were filled in capsules with filler (Psylliul). The capsules prepared were of 250 mg. Coenzyme Q10 - 75mg Vitamin E - 75mg. Vitamin C - 75mg Psylliul - 25mg. CLINICAL TRIAL: All the patients with the available record of diagnosis of chronic renal failure of minimum 12 week duration were prospectively studied for 4 weeks while receiving all other supportive treatment during which laboratory and clinical data were obtained to confirm the diagnosis of chronic renal failure. The study was approved by the ethics committee of human studies in our centre. After written informed consent, all patients with the diagnosis of chronic renal failure were randomized by the pharmacist to receive either antioxidant capsules or placebo capsules. The physician examining the patients and technicians analyzing the blood were blinded to treatment groups. All patients were asked to select blindly one of the cards, each enclosed in a sealed envelope and marked either group A or B. The intervention group A (n=48) received antioxidant formulation (2 cap thrice daily) and the placebo group B (n=49) received inert fibre cellulose (2 cap thrice daily, 500 mg each) for a period of 12 weeks. All other treatments such as furosemide, iron, calcium, vitamin D3, erythropoitin and blood transfusion were administered to both the groups. All patients were followed weekly for 4 weeks and then every 4 weekly for 12 weeks. Both groups were also encouraged to decrease the frequency or stop dialysis if there was an increase in urine output and decrease in serum creatinine of at least 2.0mg/dl. Study Methods: Clinical, ultrasonographic, echocardiaographic.radiologic and laboratory data were recorded at admission before entry to the study. Hypertension was defined as blood pressure > 140/90 mmHg and hypotension as systolic blood pressure pressure. Hypertension was treated when blood pressure was >140 mmHg systolic and or >90 mmHg diastolic with an attempt to maintain blood pressure 9.0mg/dl with or without other complications such as coma, acidosis and acute pulmonary oedema, heart failure and/or hyperkalemia (serum potassium > 6.0mEq/L). Laboratory data: A blood sample after at least 10 hours of fasting was drawn at entry and then 4, 8 and 12 week of follow up in the morning and was analysed for blood count, hemoglobin, urea, nitrogen, glucose, creatinine, sodium, potassium, vitamin, E and C and beta-carotene and thiobarbituric acid reactive substance (TBARS), diene conjugates and malondialdehyde (MDA) (16-20). Urine analysis and creatinine clearance were also done in all the patients. Laboratory personnel analysing the blood were blind to the treatment groups. Statistical analysis: The two-sample t-test using one-way analysis of variance and the Z score for proportions were used to measure the statistical significance between the two groups. Tukeys post-hoc test was also conducted for multiple comparisons. Only a P. Value Results: Of all 133 patients of chronic renal failure who were studied at our centre, 36 were excluded and 97 were randomized to either antioxidant (intervention group A. n=48) or placebo (control group B, n=49) group after stratification into hemodialysis and no hemodialysis group.Subjects in both groups were divided into those on hemodialysis and no dialysis for comparison. Table 1 shows data in 4 subgroups indicating that mean age, body weight and body mass index showed no significant difference between the concerned groups. The proportion of subjects with male sex, low haemoglobin, hypertension, diabetes mellitus and billateral contracted kidneys were comparable in the two subgroups. Treatment given before the trial indicated that the percentage of subjects given dialysis, furosemide, erythropoitin and blood transfusions for anaemia were comparable in the concerned subgroups. However, treatment given during the 12 week of trial period showed that the subjects given dialysis were significantly less in the antioxidant subgroup than control subgroup (12 vs 24, p Table 2 shows that baseline laboratory data were comparable in the two groups except for little higher serum creatinine and blood urea in the intervention group. Treatment with antioxidants was associated with significant reductions in serum creatinine and blood urea nitrogen with an increase in creatinine clearance and urine output in the antioxidant group than control group(Table 3). Plasma concentrations of vitamin E and C and beta-carotene were low and TBARS, malondialdehyde and diene conjugates which are indicators of oxidative damage were higher in all the subgroups without any significant group differences at entry to study compared to normal values observed in our laboratory in Indians.(8) After 12 weeks treatment, vitamin E and C and beta-carotene showed a significant increase and TBARS, malondialdehyde and diene conjugates showed a significant reduction in the antioxidant subgroups compared to control subgroups. Changes in sodium and potassium were not significant. Effective Range of Dosages of Various Agents: Table 5 shows that optimal dosages for reversing renal dysfunction in patients with chronic renal failure were: coenzyme Q10, 200 mg/day, vitamin E (600 mg/day) and vitamin C (600 mg/day). It is clear that the effective ranges of doses of these agents in a combination should be: coenzyme Q10, 150-200 mg/day. vitamin E, 500-600 mg/day, vitamin C 500-600 mg/day. In mg/kg body weight the dosages could be coenzyme Q10, 3.5, vitamin E, 10.0 mg and vitamin C, 10.0 mg day. Effect On Heart Failure: There were 16 patients in the intervention group A and 17 in control group B who had either enlargement of heart (4 vs 5) or congestive heart failure (12 vs 12) Heart dysfunction was demonstrated by two dimensional echocardiography, which was done before and after the treatment in all the patients. At baseline, left ventricular wall thickness, left ventricular mass, sphericity index, ejection fraction and end diastolic volume as well as end systolic volume showed no significant difference between the two groups. However, after 12 weeks follow up, there was a significant reduction in end diastolic and end systolic volume in the intervention group than control group. Left ventricular wall thickness and left ventricular mass showed significant decline in the intervention group compared to control group. Post treatment values in the treatment group showed significant beneficial changes in the intervention group without such effects in the control group. (Table 10). The findings suggested that treatment with an antioxidant combination of coenzyme Q10 +- vitamin E + vitamin C is beneficial in patients with heart failure. The dosages given in Table 5 are also applicable in heart failure. Discussion: This study shows that the proposed newly developed antioxidant composition with new dosages was effective and the contents have synergistic effects. Treatment with this combined antioxidant formulation containing coenzyme Q10, vitamin E and vitamin C in patients with chronic renal failure was associated with a significant decline in serum creatinine and blood urea nitrogen with an increase in creatinine clearance and urine output after 12 weeks of follow up in the antioxidant subgroups compared to control subgroups (Table 3). Treatment in patients with Heart failure was associated with significant benefit in heart function as observed clinically as well as in echocardiographic changes. Table 1: Effects of individual antioxidants in moderate doses in patients with chronic renal failure. Datum (Normal range) "Coenzyme Q10 (100 mg/day) Vitamin E (300 mg/day) Vitamin C (300 mg/day) Baseline(n=5) After 4 week Baseline(n=5) After 4 week Baseline(n=5) After 4 week Vitamin E (0.6-0.95 mg/dl) 0.61 0.67 0.64 0.66 0.63 0.64 Vitamin C (0.28-0.53 mg/dl) 0.23 0.25 0.26 0.25 0.22 0.24 Beta-carotene (18.8-29.5mg/dl) 18.5 19.1 18.6 18.8 18.6 18.7 TBARS (0.1-0.6 nmol/ml) 1.43 1.21 1.44 1.24 1.46 1.43 MDA (1.2-3.5 nmol/ml) 2.83 2.12 2.79 2.23 2.81 2.82 Diene conjugates (20-25,OD) 27.5 25.5 28.0 27.5 27.6 27.5 Blood urea nitrogenf 10-20 mg/dl) 69.5 60.2 70.1 68.2 71.0 70.8 Scrum creatininc(().2-1.1 mg/dl) 7.6 5.9 8.0 7.5 7.9 7.8 Creatinine clearance(>75ml/min) 25.5 28.2 26.4 28.0 27.0 27.4 Urine output(2.0-3.0 Litres/day) 2.2 2.4 2.1 2.2 2.0 2.0 Benefit + + Values are means IBARS - thiobarbituric acid reactive substances. MDA Malondialdehyde Table 2: Effects of individual antioxidants in higher doses in patients with chronic renal failure. Coenzyme Q10 (200 mg/day) Vitamin E .(600mg/day) Vitamin C(600 mg/day) Baseline After 4 week Baseline After 4 week Baseline After 4 week Vitamin E (mg/dl) 0.63 0.70 0.64 0.70 0.64 0.65 Vitamin C (mg/dl) 0.26 0.31 0.27 0.30 0.27 0.31 Beta-carotene (mg/dl) 19.60 21.70 19.0 20.5 19.2 19.3 TBARS (nmol/ml) 1.31 1.02 1.32 1.06 1.31 1.26 MDA (nmol/ml) 2.41 2.00 2.42 2.18 2.39 2.31 Diene conjugates (OD) 26.4 24.0 26.1 25.2 26.0 25.5 Blood urea nitrogen (mg/dl) 65.2 60.8 64.8 61.6 65.6 64.2 Serum creatinine (mg/dl) 7.4 6.4 7.0 6.4 7.2 7.1 Creatinine clearance (>75ml/day)27.5 28.5 27.2 28.4 27.1 27.5 Urine output (2 Litre/day) 2.2 2.5 2.1 2.3 2.2 2.3 Benefit ++ + Values are means Table 3: Effects of two combined antioxidant formulation in higher doses in patients with chronical renal failure Vitamin E (600 mg/day) Vitamin C (600 mg/day) Baseline After 4 week Coenzyme Q10 (206mg/day) Coenzyme QIO (200 mg/day) Vitamin E (600 mg/day) Vitamin C (600 mg/day) Baseline After 4 week Baseline After 4 week Vitamin E 0.66 0.72 Vitamin C 0.28 0.36 Beta-carotene 20.0 23.1 TBARS 1.6 1.4 MDA 2.8 2.5 Diene conjugates 28.8 26.0 Blood urea nitrogen 68.8 62.5 Serum creatinine 6.8 5.5 Creatinine clearance 28.0 30.1 Urine output 2.0 2.2 Benefit ++ 0.64 0.76 0.68 0.78 0.29 0.35 0.28 0.36 19.1 22.3 19.5 22.5 1.4 1.2 1.5 1.3 2.6 2.3 2.6 1.9 29.0 25.5 28.8 24.5 66.1 60.0 65.1 59.0 6.8 5.3 7.0 5.5 27.5 29.6 26.0 29.2 2.1 2.4 2.0 2.4 ++ + + Values are means Dosages: Coenzyme Q10, 3-4mg/kg body weight Vitamin C 9-1 2 mg'kg body weiglit. , vitamin E = 9-12 mg/kg body weight, Table 4: Effect of three combined antioxidant formulations in various doses in patients of chronic renal failure. Coenzyme Q10(200mg/day) Coenzyme Q10( 150 mg/day) Coenzyme Q10(100mg/day) Vitamin E (600 i mg/day) Vitamin E (500 mg/day) Vitamin E (300 mg/day) Vitamin C (600 mg/day) Vitamin C (500 mg/day) Vitamin C (300 mg/day) Baseline After 4 week Baseline After 4 week Baseline After 4 week Vitamin E 0.62 0.78 0.61 0.73 0.63 0.66 Vitamin C 0.20 0.38 0.19 0.31 0.21 0.25 Beta-carotene 17.5 25.5 18.6 21.6 19.0 20.5 TBARS 1.52 1.0 1.55 1.31 1.56 1.42 MDA 2.86 1.6 2.91 2.41 2.92 2.67 Diene conjugates 29.5 24.2 29.8 26.5 29.0 27.5 Blood urea nitrogen 70.5 60.1 71.0 65.4 69.5 68.5 Serum creatinine 7.8 5.2 7.7 6.7 7.6 7.0 Creatinine clearance 24.4 31.2 23.8 24.8 23.0 23.5 Urine output 1.9 2.4 1.8 2.0 1.8 1.9 Benefit +++ ++ + Values are means. Table 5: Effective range of dosages of various agents used in the new formulation. Agents Total dose Dosage schedule Content Percentage mg/kg Effective (mg/kg) thrice daily per of each agent body range (mg) capsule(mg) per capsule weight (mg/day) Coenzyme Q10 200 66.6 33.3 14.3 3.5 150-200 Vitamin E 600 200 100 42.8 10.0 500-600 Vitamin C 600 200 100 42.8 10.0 500-600 Total Quantity 1400 466.6 233.3 99.9 TabIe-6: Clinical characteristics of subjects in the coenzyme QIO and control groups. Datum Hemodialysis No dialysis Hemodialysis No dialysis (n=21) (n=27) (n=24) (n=25) Mean age (years) 51.5±11.8 46.8+12.4 48.4+13.1 46.3+11.7 Body weight (kg.) 57.0+9.6 58.8±13.6 56.0+11.5 56.8+12.3 Body Mass intake (kg/m2) 21.0+3.0 n(%) 22.3±3.1 20.0+2.4 n(%) 20.8+3.0 Men 16(76.2) 18(66.6) 17(70.8) 18(72.0) Low haemoglobin ( Hypertension (> 140/90 mmHg) 19(90.4) 14(51.8) 18(75.0) 20(80.0) Diabetes mellitus (known) 8(38.1) 7(25.9) 9(37.5) 9(36.0) Billateral contracted kidneys 20(95.2) 23(85.2) 21(87.5) 22(88.0) (Ultrasonography) Etilogy of renal failure Diabetes mellitus 8(38.1) 9(33.3) 8(33.3) 9(36.0) Hypertension 3(14.2) 2(7.4) 3(12.5) 2(8.0) Chronic glomerulonephritis 6(28.5) 9(33.3) 9(37.5) 9(36.0) Chronic pyeloephritis 2(9.5) 5(18.5) 2(8.3) 3(12.0) Unknown 2(9.5) 2(7.4) 2(8.3) 2(8.0) Treatment receiving before trial. Total number of dialysis in the last one week before entry. 46(219) — 54(225) — Furosemide(40-120mg/day) 5(23.8) 24(88.8) 3(12.5) 25(100) Blood transfusion 17(80.9) 16(59.2) 22(91.6) 14(56.0) Erythropointin 20(95.2) 13(48.1) 22(91.6) 14(56.0) (2000-4000 units/week) Treatment given during the trial. Total number of dialysis in the 24(114)* — 62(258) — last one week after entry. Hemodialysis 12(57.0)* — 24(100) — Furosemide(40-120mg/dl) 20(95.2) 25(92.6) 20(83.2) 25(100) Blood transfusion 20(95.2) 25(92.6) 21(87.5) 15(60.0) Erythropointin 19(90.4) 13(48.1) 23(95.8) 15(60.0) (2000-4000 units/week) * = p Table-7: Biochemical data in the antioxidant and control group at baseline. Datum Antioxidant group (n=48) Hemodialysis No dialysis (n=21) (n=27) Control group (n=49) Hemodialysis No dialysis (n=24) (n=25) Vitamin E (0.6-0.95 mg/dl) 0.71+0.07 Vitamin C (0.28-0.53 mg/dl) 0.29+0.06 Beta-carotene (18.8-29.5ng/dl) 21.0+3.7 TBARS (0.1-0.6 nmol/ml) 1.5+0.28 Malondialdehyde (3.6-8.7 nmol/ml) 2.9+0.48 Diene conjugates(20-25. OD units) 31.0+3.2 Blood urea nitrogen (10-20 mg/dl) 88.2+26.0* Scrum creatinine (0.2-1.1 mg/dl) 9.5+2.3* Creatinine clearance (>75ml/min) 40.0+10.9 Urine output (2.0-2.5 litres/24 hours) 1.3+0.37 0.70+0.04 0.28+0.07 22.8±4.0 1.49+0.33 2.9±0.6 31.6±3.7 67.5+30.6 7.1+2.6 42.9±9.5 1.4+0.42 0.67+0.03 0.28+0.07 18.2+3..23 1.7±0.34 3.1±0.48 31.9+2.4 95.1±42.5* 10.8+1.8* 38.2+12.8 1.3+0.16 0.64+0.05 0.25+0.05 19.4+3.94 1.7+0.28 3.2+0.4 34.1+6.7 66.9+17.8 6.9+2.05 42.0+29.6 1.5+0.15 Values are mean + standard deviation. * = P Table 8: Biochemical data in the antioxidant and control subgroups after treatment. Datum Antioxidant Hemodialysis (n=2!) group No dialysis (n=27) Control group Hemodialysis No dialysis (n=24) (n=25) Changes (Antioxidant) Hemodialysis No hemodialysis (n=21) (n=27) Changes (Control) Hemodialysis No hemodialysis (n=24) (n=25) 0.73+0.03* 0.33+0.05* 24 + 5.7* 1.3+0.3* 89+27.88 1.44+0.17 ■ Vitamin E (mg/dl) Vitamin C (mg/dl) Beta-carotene (ug/dl) TBARS (nmol/ml) Malondialdehyde(nmol/ml)l .9+0.48* Diene conjugates 26+2.2* (OD units) Blood urea nitrogen 79.8+2 6.82 (mg/dl) Serum creatinine 6.7+1.6* (mg/dl) 54.9+9.6* Creatinine clearance (ml/min) 1.92+0.35* Urine output (Litre/24 hours) 0.73+0.03* 0.33+0.03* 24.9+3.5* 1.1+0.3* 2.0+0.6* 25.7+3.9* 63.5+15.12 5.4+2.1* 53.6+8.41 1.75+0.38* 0.64+0.04 0.58+0.05 0.02+0.041 0.20+0.07 0.23+0.06 0.04+0.05 16.3+ 4.66 17.1+4.91 03+4.22* 1.8+0.37 1.8+0.30 -0.2+0.31 2.9+0.65 3.2±5.3 -01+0.31* 33.7+4.25 30.2+4.37 -05+2.20* 80.1+32.6 -8.4+31.10* 11.2+1.88 8.0+2.79 -2.8+2.19* 30.7+16.76 36.2+24.16 45.4+7.51* 1.35+0.28 0.36+0.33' 0.03+0.04* -0.03±0.05 -0.06+0.05 0.05+0.06* -0.08+0.07 0.02+0.04 2.1+3.46' -1.9+3.41 -2.3+4.2 -0.39+0.4* 0.12+0.26 0.1+0.21 -0.9±0.28* -0.2±.I1 0.16+0.09 -5.9+2.10* 1.83+2.63 -1.76+2.03 -4.0+18.62 83.2+33.7 13.2 + 22.3 -1.7+1.61* 0.5+1.23 l.t+1.60 10.7+4.9* -7.5+4.8 -5.8+413 0.35+0.31* 0.04±0.14 -0.1+0.26 Values are mean+ standard deviation, * = P Table 9: Adverse effects of antioxidant formulation. Manifestations Antioxidant group Control group Hemodialysis No dialysis Hemodialysis No dialysis (n=21) (n=27) (n-24) (n=25 Fulness Nausea Vomiting Loose stools Headache Bodyache Palpitation 4(19.0) 2(7!4) 5(20.8) 2(8.0) 7(33.3) 5(18.5) 8(33.3) 4(16.0) 2(9.5) 2(7.4) 3(12.5) 2(8.0) 4(19.0) 2(7.4) 1(4.7) 1(4.1) 2(9.5) - 1(4.1) Table 10: Changes in left ventricular function on two-dimensional echocardiography. Coenzyme Q10 group (n=16) Control group (n: =17) Baseline After treatment Baseline After treatment Left ventricular 13.4±2.7 11.2+1.8* 14.1+3.1 14.8+3.3 wall thickness(mm) Left ventricular 242+70 205+52* 245±75 254+78 mass (g) Sphericity index 1.62±0.25 1.63±0.28 1.62+0.31 1.34+0.18* Ejection fraction 47.5±5.6 55.2+6.5* 45.6+4.2 48.2+5.3 End diastolic 108.2+3.2 95.5+1.8* 107.5±3.0 105.2+2.8 Volume(ml) End systolic volume(ml) 62.6+1.6 53.4+1.5* 63.0+1.7 61.4+1.6 * =P - fable 10: Changes in left ventricular function on two-dimensional echocardiography. Coenzyme Q10 group (n=16) Control group (n= 17) Baseline After treatment Baseline After treatment Left ventricular 13.4±2.7 11.2±1.8* 14.1±3.1 14.8±3.3 wall thickness(mm) Left ventricular 242±70 205+52* 245+75 254+78 mass (g) Sphericity index 1.62.+0.25 1.63±0.28 1.62+0.31 1.34±0.18* Ejection fraction 47.5±5.6 55.2±6.5* 45.6±4.2 48.2±5.3 End diastolic 108.2+3.2 95.5+1.8* 107.5±3.0 105.2±2.8 Volume(ml) End systolic volume(ml) 62.6+1.6 53.4±1.5* 63.0+1.7 61.4+1.6 * =P Conclusion: Independent action of these anti-oxidants was modest but non-significant, whereas in combination the action was good indicating synergistic action of these molecules. It is possible that the synergistic effect due to bio-energetic and ATP (adenosine tri phosphotase) producing activity of coenzyme Q10. The activity of coenzyme Q10 is potentiated by the activity of Vitamin E and Vitamin C. Coenzyme Q10 also has a sparing effect on intracellular levels of Vitamin E since Q 10 has multiple beneficiary activities it should have decreased the anti-oxidants conjunction by the cells of the body and potentiated the overall composition. We Claim: 1. A process of preparing a new antioxidant composition for the treatment of chronic renal failure and heart failure comprising mixing the following anti-oxidants in the proportions indicated there against: Coenzyme Q10 (Ubiquinone) - 10-40% by wt. Vitamin E - 30-45% by wt. Vitamin C - 30-45% by wt. and the balance if any adding a conventional filler fof preparing the tablet or capsule. 2. A process as claimed in claim 1 wherein said filler used is Psylliul. 3. A process as claimed in claim 1 wherein the proportions of the ingredients in the anti-oxidant composition are as follows Coenzyme Q10 - 20% by wt. Vitamin E - 40%bywt. Vitamin C - 40% by wt. 4. A process as claimed in claim 1 wherein the proportions of the ingredients in the anti-oxidant composition are as follows Coenzyme Q10 - 30%bywt. Vitamin E - 30% by wt. Vitamin C - 30% by wt. Filler - 10%bywt. 5. A process of preparing a new anti-oxidant composition for the treatment of chronic renal failure and heart failure substantially as herein described with reference to the foregoing examples. Dated this 12th day of July, 2000 of Anand & Anand, Advocates Agents for the Applicants

Full Text

FORM-3 A
THE PATENTS ACT, 1970
i
COMPLETE SPECIFICATION
SECTION 10
Title:- A PROCESS OF PREPARING A NEW ANTI-OXIDANT COMPOSITION
FOR THE TREATMENT OF CHRONIC RENAL FAILURE AND HEART FAILURE,
Applicant (8): - PROF, RAM BHADUR SINGH AND
DR.MOHAMMAD ARIZ NIAZ,
SAFE ENTERPRISES, 6 CITIZEN APARTMENT 30TH ROAD, BANDRA, MUMBAI - 400 050, •MAHARASHTRA, INDIA.
The following Specification particularly deadbea and ascertains the nature of this Invention and the manner in which it is to be performed :-

This invention relates to a process of preparing a new anti-oxidant composition for the treatment of chronic renal failure and heart failure.
BACKGROUND
Superoxide and hydroxyl radicals and hydrogen peroxide are important reactive oxygen species in the pathogenesis of renal damage resulting into renal failure.(l-4) Evidence supporting a role for oxidative stress in renal injury has been derived predominantly from studies using a variety of antioxidants such as superoxide dismutase,glutathione peroxidase and catalase.(2-7).Case control studies (8,9) indicate that there is deficiency of antioxidant vitamins A, E and C and beta-carotene as well as coenzyme Q10, lycopene, zinc, selenium in patients with chronic renal failure with and without dialysis as well as in heart failure. However, all studies have not reported a deficiency of antioxidants in renal failure and heart failure. Increased levels of thiobarbituric acid reactive substances (TBARS), and malondialdehyde (MDA) which are indicators of free radical damage have also been described in patients with chronic renal failure and heart failure. However, no study has examined the role of antioxidants in patients with chronic renal failure associated with heart failure. The role of coenzyme Q10 in heart failure is known however the dosages and Efficacy is controversial.
The object of this invention is to examine the role of anti-oxidants in patients with chronic renal failure and in a substudy to find out whether it is also beneficial in heart failure and thereafter develop a composition of anti-oxidants for the treatment of chronic renal failure and heart failure.
To achieve the said objective, the effects of individual anti-oxidants in moderate, higher, in combination were studied by administering various anti-oxidants in randomized, double blind trials without a control group for a period of four weeks to study the synergistic effects, as given in tables 1-5. A randomized, double blind and controlled trial was conducted in 97 patients to confirm our findings both in renal failure and heart failure (Tables 6-10).

Effects of individual antioxidants in moderate doses:
We administered coenzyme Q10 (100 mg/day), vitamin E (300 mg/day) and vitamin C(300 mg/day) after randomizing the subjects into three groups (Table 1). There was a modest reduction in serum creatinine and blood urea in the coenzyme Q10 group, doubtful beneficial effect in the vitamin E group and no effect in vitamin C group. The changes were non-significant.
Effects of individual antioxidants in higher doses:
Subjects were randomized in to three groups in a double blind fashion. They were administered, coenzyme QIO (200 mg/day) or vitamin E (600 mg/day) or vitamin C (600 mg/day) in the concerned group for a period of 4 weeks. Treatment with coenzyme Q10 was associated with substantial reduction in blood urea nitrogen and serum creatinine and increase in creatinine clearance and urine output (Table 2). Treatment with vitamin E also caused substantial changes but slightly lower than coenzyme Q10. Vitamin C had neutral effect. There was significant beneficial effect in patients with heart failure in the coenzume Q10 group with little good effect in vitamin E group and no effect in vitamin C group.
Effects of two combined antioxidant formulation in higher doses:
Subjects were randomized into three groups (after a wash out period of 4 weeks) in a double blind fashion. They were administered coenzyme QlO+Vitamin E, Coenzyme QlO+Vitamin C and vitamin E + Vitamin C. There was a substantial reduction in blood urea nitrogen and serum creatinine and an increase in creatinine clearance and urine output indicating that some synergistic effect is possible. Patients with heart failure (2 in each group) showed substantial beneficial effect in the first two group and non-significant benefit in the third group.

Effect of three combined antioxidant formulations in various doses:
Subjects were randomized into three groups as given in table 4 and were administered various combinations of formulations in a double blind fashion for 4 weeks. High dose combination of coenzyme Q10 (200 mg) + vitamin E (600 mg) + vitamin C (600 mg) per day showed a marked reduction in blood urea nitrogen and serum creatinine and an increase in creatinine clearance and urine output as well as in heart function. Moderate dose combination also caused substantial improvement in renal function, and heart function (in patients with heart failure) but lower compared to high dose combination. However lowest dose combination showed only mild nonsignificant benefit in renal function indicating failure of this dosage schedule in reversing renal dysfunction. High and moderate dose groups also caused significant benefit in heart function in patients with heart failure. Finally the results with high dose combination were examined in a randomized, controlled trial in 97 patients of chronic renal failure including 33 with heart failure (Table 6-10).There was a significant benefit in patients with renal failure as well as in patients with heart failure(Table 10).Side effects were mild(Table 9).
In view of the above study, a process for preparing a new anti-oxidant composition for the treatment of chronic renal failure and heart failure was developed. According to this invention, the said process of preparing a new anti-oxidant composition for the treatment of chronic renal failure and heart failure comprising mixing the following anti-oxidants in the proportion indicated there against:
Coenzyme Q10(Ubiquinone)- 10-40% by wt.
Vitamin E - 30-45% by wt.
Vitamin C - 30-45% by wt.
and the balance if any adding conventional fillers for preparing the tablet or capsule. The filler is used to make the quantity of the composition acceptable and also to fill the capsule. The said filler used is Psvlliul.

The proportions of the ingredients used in the preparation of anti-oxidant
composition are as follows
Coenzyme QIO - 20%bywt.
Vitamin E - 40% by wt.
Vitamin C - 40%bywt.
The proportions of the ingredients in the preparation of anti-oxidant composition are
as follows ,
Coenzyme QIO - 30%bywt.
Vitamin E - 30% by wt.
Vitamin C - 30% by wt.
Filler - 10%bywt.
The invention will now be described with reference to the following examples;
Example 1:
The test agent anti-oxidants, coenzymes QIO, Vitamin C and vitamin E were purchased from the market. These ingredients were mixed in the following proportions and thereafter they were filled in capsules without filler. The capsules prepared were of 250 mg.
Coenzyme QIO - 50mg.
Vitamin E - lOOmg.
Vitamin C - lOOmg
Example 2: .
The test agent anti-oxidants, coenzymes QIO, Vitamin C and vitamin E were purchased from the market. These ingredients were mixed in the following proportions and thereafter they were filled in capsules with filler (Psylliul). The capsules prepared were of 250 mg.

Coenzyme Q10 - 75mg
Vitamin E - 75mg.
Vitamin C - 75mg
Psylliul - 25mg.
CLINICAL TRIAL:
All the patients with the available record of diagnosis of chronic renal failure of minimum 12 week duration were prospectively studied for 4 weeks while receiving all other supportive treatment during which laboratory and clinical data were obtained to confirm the diagnosis of chronic renal failure. The study was approved by the ethics committee of human studies in our centre. After written informed consent, all patients with the diagnosis of chronic renal failure were randomized by the pharmacist to receive either antioxidant capsules or placebo capsules. The physician examining the patients and technicians analyzing the blood were blinded to treatment groups. All patients were asked to select blindly one of the cards, each enclosed in a sealed envelope and marked either group A or B. The intervention group A (n=48) received antioxidant formulation (2 cap thrice daily) and the placebo group B (n=49) received inert fibre cellulose (2 cap thrice daily, 500 mg each) for a period of 12 weeks. All other treatments such as furosemide, iron, calcium, vitamin D3, erythropoitin and blood transfusion were administered to both the groups. All patients were followed weekly for 4 weeks and then every 4 weekly for 12 weeks. Both groups were also encouraged to decrease the frequency or stop dialysis if there was an increase in urine output and decrease in serum creatinine of at least 2.0mg/dl.
Study Methods:
Clinical, ultrasonographic, echocardiaographic.radiologic and laboratory data were recorded at admission before entry to the study. Hypertension was defined as blood pressure > 140/90 mmHg and hypotension as systolic blood pressure
pressure. Hypertension was treated when blood pressure was >140 mmHg systolic and or >90 mmHg diastolic with an attempt to maintain blood pressure 9.0mg/dl with or without other complications such as coma, acidosis and acute pulmonary oedema, heart failure and/or hyperkalemia (serum potassium > 6.0mEq/L).
Laboratory data:
A blood sample after at least 10 hours of fasting was drawn at entry and then 4, 8 and 12 week of follow up in the morning and was analysed for blood count, hemoglobin, urea, nitrogen, glucose, creatinine, sodium, potassium, vitamin, E and C and beta-carotene and thiobarbituric acid reactive substance (TBARS), diene conjugates and malondialdehyde (MDA) (16-20). Urine analysis and creatinine clearance were also done in all the patients. Laboratory personnel analysing the blood were blind to the treatment groups.
Statistical analysis:
The two-sample t-test using one-way analysis of variance and the Z score for proportions were used to measure the statistical significance between the two groups. Tukeys post-hoc test was also conducted for multiple comparisons. Only a P. Value Results:
Of all 133 patients of chronic renal failure who were studied at our centre, 36 were excluded and 97 were randomized to either antioxidant (intervention group A. n=48)

or placebo (control group B, n=49) group after stratification into hemodialysis and no hemodialysis group.Subjects in both groups were divided into those on hemodialysis and no dialysis for comparison. Table 1 shows data in 4 subgroups indicating that mean age, body weight and body mass index showed no significant difference between the concerned groups. The proportion of subjects with male sex, low haemoglobin, hypertension, diabetes mellitus and billateral contracted kidneys were comparable in the two subgroups. Treatment given before the trial indicated that the percentage of subjects given dialysis, furosemide, erythropoitin and blood transfusions for anaemia were comparable in the concerned subgroups. However, treatment given during the 12 week of trial period showed that the subjects given dialysis were significantly less in the antioxidant subgroup than control subgroup (12 vs 24, p Table 2 shows that baseline laboratory data were comparable in the two groups except for little higher serum creatinine and blood urea in the intervention group. Treatment with antioxidants was associated with significant reductions in serum creatinine and blood urea nitrogen with an increase in creatinine clearance and urine output in the antioxidant group than control group(Table 3).
Plasma concentrations of vitamin E and C and beta-carotene were low and TBARS, malondialdehyde and diene conjugates which are indicators of oxidative damage

were higher in all the subgroups without any significant group differences at entry to study compared to normal values observed in our laboratory in Indians.(8) After 12 weeks treatment, vitamin E and C and beta-carotene showed a significant increase and TBARS, malondialdehyde and diene conjugates showed a significant reduction in the antioxidant subgroups compared to control subgroups. Changes in sodium and potassium were not significant.
Effective Range of Dosages of Various Agents:
Table 5 shows that optimal dosages for reversing renal dysfunction in patients with chronic renal failure were: coenzyme Q10, 200 mg/day, vitamin E (600 mg/day) and vitamin C (600 mg/day). It is clear that the effective ranges of doses of these agents in a combination should be: coenzyme Q10, 150-200 mg/day. vitamin E, 500-600 mg/day, vitamin C 500-600 mg/day. In mg/kg body weight the dosages could be coenzyme Q10, 3.5, vitamin E, 10.0 mg and vitamin C, 10.0 mg day.
Effect On Heart Failure:
There were 16 patients in the intervention group A and 17 in control group B who had either enlargement of heart (4 vs 5) or congestive heart failure (12 vs 12) Heart dysfunction was demonstrated by two dimensional echocardiography, which was done before and after the treatment in all the patients.
At baseline, left ventricular wall thickness, left ventricular mass, sphericity index, ejection fraction and end diastolic volume as well as end systolic volume showed no significant difference between the two groups. However, after 12 weeks follow up, there was a significant reduction in end diastolic and end systolic volume in the intervention group than control group. Left ventricular wall thickness and left ventricular mass showed significant decline in the intervention group compared to control group. Post treatment values in the treatment group showed significant beneficial changes in the intervention group without such effects in the control group. (Table 10).

The findings suggested that treatment with an antioxidant combination of coenzyme Q10 +- vitamin E + vitamin C is beneficial in patients with heart failure. The dosages given in Table 5 are also applicable in heart failure.
Discussion:
This study shows that the proposed newly developed antioxidant composition with new dosages was effective and the contents have synergistic effects. Treatment with this combined antioxidant formulation containing coenzyme Q10, vitamin E and vitamin C in patients with chronic renal failure was associated with a significant decline in serum creatinine and blood urea nitrogen with an increase in creatinine clearance and urine output after 12 weeks of follow up in the antioxidant subgroups compared to control subgroups (Table 3).
Treatment in patients with Heart failure was associated with significant benefit in heart function as observed clinically as well as in echocardiographic changes.

Table 1: Effects of individual antioxidants in moderate doses in patients with chronic renal failure.
Datum (Normal range) "Coenzyme Q10 (100 mg/day) Vitamin E (300 mg/day) Vitamin C (300 mg/day)
Baseline(n=5) After 4 week Baseline(n=5) After 4 week Baseline(n=5) After 4 week

Vitamin E (0.6-0.95 mg/dl) 0.61 0.67 0.64 0.66 0.63 0.64
Vitamin C (0.28-0.53 mg/dl) 0.23 0.25 0.26 0.25 0.22 0.24
Beta-carotene (18.8-29.5mg/dl) 18.5 19.1 18.6 18.8 18.6 18.7
TBARS (0.1-0.6 nmol/ml) 1.43 1.21 1.44 1.24 1.46 1.43
MDA (1.2-3.5 nmol/ml) 2.83 2.12 2.79 2.23 2.81 2.82
Diene conjugates (20-25,OD) 27.5 25.5 28.0 27.5 27.6 27.5
Blood urea nitrogenf 10-20 mg/dl) 69.5 60.2 70.1 68.2 71.0 70.8
Scrum creatininc(().2-1.1 mg/dl) 7.6 5.9 8.0 7.5 7.9 7.8
Creatinine clearance(>75ml/min) 25.5 28.2 26.4 28.0 27.0 27.4
Urine output(2.0-3.0 Litres/day) 2.2 2.4 2.1 2.2 2.0 2.0
Benefit + +
Values are means IBARS - thiobarbituric acid reactive substances. MDA Malondialdehyde

Table 2: Effects of individual antioxidants in higher doses in patients with chronic renal failure.
Coenzyme Q10 (200 mg/day) Vitamin E .(600mg/day) Vitamin C(600 mg/day)
Baseline After 4 week Baseline After 4 week Baseline After 4 week

Vitamin E (mg/dl) 0.63 0.70 0.64 0.70 0.64 0.65
Vitamin C (mg/dl) 0.26 0.31 0.27 0.30 0.27 0.31
Beta-carotene (mg/dl) 19.60 21.70 19.0 20.5 19.2 19.3
TBARS (nmol/ml) 1.31 1.02 1.32 1.06 1.31 1.26
MDA (nmol/ml) 2.41 2.00 2.42 2.18 2.39 2.31
Diene conjugates (OD) 26.4 24.0 26.1 25.2 26.0 25.5
Blood urea nitrogen (mg/dl) 65.2 60.8 64.8 61.6 65.6 64.2
Serum creatinine (mg/dl) 7.4 6.4 7.0 6.4 7.2 7.1
Creatinine clearance (>75ml/day)27.5 28.5 27.2 28.4 27.1 27.5
Urine output (2 Litre/day) 2.2 2.5 2.1 2.3 2.2 2.3
Benefit ++ +
Values are means

Table 3: Effects of two combined antioxidant formulation in higher doses in patients with chronical renal failure

Vitamin E (600 mg/day)
Vitamin C (600 mg/day)
Baseline After 4 week

Coenzyme Q10 (206mg/day) Coenzyme QIO (200 mg/day)
Vitamin E (600 mg/day) Vitamin C (600 mg/day)
Baseline After 4 week Baseline After 4 week

Vitamin E
0.66 0.72
Vitamin C 0.28 0.36
Beta-carotene 20.0 23.1
TBARS 1.6 1.4
MDA 2.8 2.5
Diene conjugates 28.8 26.0
Blood urea nitrogen 68.8 62.5
Serum creatinine 6.8 5.5
Creatinine clearance 28.0 30.1
Urine output 2.0 2.2
Benefit
++

0.64 0.76 0.68 0.78
0.29 0.35 0.28 0.36
19.1 22.3 19.5 22.5
1.4 1.2 1.5 1.3
2.6 2.3 2.6 1.9
29.0 25.5 28.8 24.5
66.1 60.0 65.1 59.0
6.8 5.3 7.0 5.5
27.5 29.6 26.0 29.2
2.1 2.4 2.0 2.4
++ + +

Values are means Dosages: Coenzyme Q10, 3-4mg/kg body weight Vitamin C 9-1 2 mg'kg body weiglit.

, vitamin E = 9-12 mg/kg body weight,

Table 4: Effect of three combined antioxidant formulations in various doses in patients of chronic renal failure.

Coenzyme Q10(200mg/day) Coenzyme Q10( 150 mg/day) Coenzyme Q10(100mg/day)
Vitamin E (600 i mg/day) Vitamin E (500 mg/day) Vitamin E (300 mg/day)
Vitamin C (600 mg/day) Vitamin C (500 mg/day) Vitamin C (300 mg/day)
Baseline After 4 week Baseline After 4 week Baseline After 4 week
Vitamin E 0.62 0.78 0.61 0.73 0.63 0.66
Vitamin C 0.20 0.38 0.19 0.31 0.21 0.25
Beta-carotene 17.5 25.5 18.6 21.6 19.0 20.5
TBARS 1.52 1.0 1.55 1.31 1.56 1.42
MDA 2.86 1.6 2.91 2.41 2.92 2.67
Diene conjugates 29.5 24.2 29.8 26.5 29.0 27.5
Blood urea nitrogen 70.5 60.1 71.0 65.4 69.5 68.5
Serum creatinine 7.8 5.2 7.7 6.7 7.6 7.0
Creatinine clearance 24.4 31.2 23.8 24.8 23.0 23.5
Urine output 1.9 2.4 1.8 2.0 1.8 1.9
Benefit +++ ++ +
Values are means.

Table 5: Effective range of dosages of various agents used in the new formulation.
Agents Total dose Dosage schedule Content Percentage mg/kg Effective
(mg/kg) thrice daily per of each agent body range
(mg) capsule(mg) per capsule weight (mg/day)
Coenzyme Q10 200 66.6 33.3 14.3 3.5 150-200
Vitamin E 600 200 100 42.8 10.0 500-600
Vitamin C 600 200 100 42.8 10.0 500-600
Total Quantity 1400 466.6 233.3 99.9

TabIe-6: Clinical characteristics of subjects in the coenzyme QIO and control groups.

Datum Hemodialysis No dialysis Hemodialysis No dialysis
(n=21) (n=27) (n=24) (n=25)
Mean age (years) 51.5±11.8 46.8+12.4 48.4+13.1 46.3+11.7
Body weight (kg.) 57.0+9.6 58.8±13.6 56.0+11.5 56.8+12.3
Body Mass intake (kg/m2) 21.0+3.0
n(%) 22.3±3.1 20.0+2.4 n(%) 20.8+3.0
Men 16(76.2) 18(66.6) 17(70.8) 18(72.0)
Low haemoglobin ( Hypertension (> 140/90 mmHg) 19(90.4) 14(51.8) 18(75.0) 20(80.0)
Diabetes mellitus (known) 8(38.1) 7(25.9) 9(37.5) 9(36.0)
Billateral contracted kidneys 20(95.2) 23(85.2) 21(87.5) 22(88.0)
(Ultrasonography)
Etilogy of renal failure
Diabetes mellitus 8(38.1) 9(33.3) 8(33.3) 9(36.0)
Hypertension 3(14.2) 2(7.4) 3(12.5) 2(8.0)
Chronic glomerulonephritis 6(28.5) 9(33.3) 9(37.5) 9(36.0)
Chronic pyeloephritis 2(9.5) 5(18.5) 2(8.3) 3(12.0)
Unknown 2(9.5) 2(7.4) 2(8.3) 2(8.0)
Treatment receiving before trial.
Total number of dialysis in the
last one week before entry. 46(219) — 54(225) —
Furosemide(40-120mg/day) 5(23.8) 24(88.8) 3(12.5) 25(100)
Blood transfusion 17(80.9) 16(59.2) 22(91.6) 14(56.0)
Erythropointin 20(95.2) 13(48.1) 22(91.6) 14(56.0)
(2000-4000 units/week)
Treatment given during the trial.
Total number of dialysis in the 24(114)* — 62(258) —
last one week after entry.
Hemodialysis 12(57.0)* — 24(100) —
Furosemide(40-120mg/dl) 20(95.2) 25(92.6) 20(83.2) 25(100)
Blood transfusion 20(95.2) 25(92.6) 21(87.5) 15(60.0)
Erythropointin 19(90.4) 13(48.1) 23(95.8) 15(60.0)
(2000-4000 units/week)
* = p
Table-7: Biochemical data in the antioxidant and control group at baseline.

Datum

Antioxidant group (n=48)
Hemodialysis No dialysis
(n=21) (n=27)

Control group (n=49)
Hemodialysis No dialysis
(n=24) (n=25)

Vitamin E (0.6-0.95 mg/dl) 0.71+0.07
Vitamin C (0.28-0.53 mg/dl) 0.29+0.06
Beta-carotene (18.8-29.5ng/dl) 21.0+3.7
TBARS (0.1-0.6 nmol/ml) 1.5+0.28
Malondialdehyde (3.6-8.7 nmol/ml) 2.9+0.48
Diene conjugates(20-25. OD units) 31.0+3.2
Blood urea nitrogen (10-20 mg/dl) 88.2+26.0*
Scrum creatinine (0.2-1.1 mg/dl) 9.5+2.3*
Creatinine clearance (>75ml/min) 40.0+10.9
Urine output (2.0-2.5 litres/24 hours) 1.3+0.37

0.70+0.04
0.28+0.07
22.8±4.0
1.49+0.33
2.9±0.6
31.6±3.7
67.5+30.6
7.1+2.6
42.9±9.5
1.4+0.42

0.67+0.03 0.28+0.07 18.2+3..23 1.7±0.34 3.1±0.48 31.9+2.4 95.1±42.5* 10.8+1.8* 38.2+12.8 1.3+0.16

0.64+0.05
0.25+0.05
19.4+3.94
1.7+0.28
3.2+0.4
34.1+6.7
66.9+17.8
6.9+2.05
42.0+29.6
1.5+0.15

Values are mean + standard deviation. * = P
Table 8: Biochemical data in the antioxidant and control subgroups after treatment.

Datum

Antioxidant Hemodialysis (n=2!)

group No dialysis (n=27)

Control group
Hemodialysis No dialysis
(n=24) (n=25)

Changes (Antioxidant)
Hemodialysis No hemodialysis
(n=21) (n=27)

Changes (Control)
Hemodialysis No hemodialysis
(n=24) (n=25)

0.73+0.03* 0.33+0.05* 24 + 5.7* 1.3+0.3*
89+27.88
1.44+0.17
■ Vitamin E (mg/dl) Vitamin C (mg/dl) Beta-carotene (ug/dl) TBARS (nmol/ml)
Malondialdehyde(nmol/ml)l .9+0.48*
Diene conjugates 26+2.2*
(OD units)
Blood urea nitrogen 79.8+2 6.82
(mg/dl)
Serum creatinine
6.7+1.6*
(mg/dl)
54.9+9.6*
Creatinine clearance
(ml/min)
1.92+0.35*
Urine output
(Litre/24 hours)

0.73+0.03*
0.33+0.03*
24.9+3.5*
1.1+0.3*
2.0+0.6*
25.7+3.9*
63.5+15.12
5.4+2.1*
53.6+8.41
1.75+0.38*

0.64+0.04 0.58+0.05 0.02+0.041
0.20+0.07 0.23+0.06 0.04+0.05
16.3+ 4.66 17.1+4.91 03+4.22*
1.8+0.37 1.8+0.30 -0.2+0.31
2.9+0.65 3.2±5.3 -01+0.31*
33.7+4.25 30.2+4.37 -05+2.20*
80.1+32.6 -8.4+31.10*
11.2+1.88
8.0+2.79 -2.8+2.19*
30.7+16.76 36.2+24.16 45.4+7.51*
1.35+0.28 0.36+0.33'

0.03+0.04* -0.03±0.05 -0.06+0.05
0.05+0.06* -0.08+0.07 0.02+0.04
2.1+3.46' -1.9+3.41 -2.3+4.2
-0.39+0.4* 0.12+0.26 0.1+0.21
-0.9±0.28* -0.2±.I1 0.16+0.09
-5.9+2.10* 1.83+2.63 -1.76+2.03
-4.0+18.62 83.2+33.7 13.2 + 22.3
-1.7+1.61* 0.5+1.23 l.t+1.60
10.7+4.9* -7.5+4.8 -5.8+413
0.35+0.31* 0.04±0.14 -0.1+0.26

Values are mean+ standard deviation, * = P
Table 9: Adverse effects of antioxidant formulation.

Manifestations Antioxidant group Control group
Hemodialysis No dialysis Hemodialysis No dialysis
(n=21) (n=27) (n-24) (n=25
Fulness
Nausea
Vomiting
Loose stools
Headache
Bodyache
Palpitation
4(19.0) 2(7!4) 5(20.8) 2(8.0)
7(33.3) 5(18.5) 8(33.3) 4(16.0)
2(9.5) 2(7.4) 3(12.5) 2(8.0)
4(19.0) 2(7.4)
1(4.7)
1(4.1)
2(9.5) - 1(4.1)

Table 10: Changes in left ventricular function on two-dimensional echocardiography.

Coenzyme Q10 group (n=16) Control group (n: =17)
Baseline After treatment Baseline After treatment
Left ventricular 13.4±2.7 11.2+1.8* 14.1+3.1 14.8+3.3
wall thickness(mm)
Left ventricular 242+70 205+52* 245±75 254+78
mass (g)
Sphericity index 1.62±0.25 1.63±0.28 1.62+0.31 1.34+0.18*
Ejection fraction 47.5±5.6 55.2+6.5* 45.6+4.2 48.2+5.3
End diastolic 108.2+3.2 95.5+1.8* 107.5±3.0 105.2+2.8
Volume(ml)
End systolic volume(ml) 62.6+1.6 53.4+1.5* 63.0+1.7 61.4+1.6
* =P
- fable 10: Changes in left ventricular function on two-dimensional echocardiography.
Coenzyme Q10 group (n=16) Control group (n= 17)
Baseline After treatment Baseline After treatment

Left ventricular 13.4±2.7 11.2±1.8* 14.1±3.1 14.8±3.3
wall thickness(mm)
Left ventricular 242±70 205+52* 245+75 254+78
mass (g)
Sphericity index 1.62.+0.25 1.63±0.28 1.62+0.31 1.34±0.18*
Ejection fraction 47.5±5.6 55.2±6.5* 45.6±4.2 48.2±5.3
End diastolic 108.2+3.2 95.5+1.8* 107.5±3.0 105.2±2.8
Volume(ml)
End systolic volume(ml) 62.6+1.6 53.4±1.5* 63.0+1.7 61.4+1.6
* =P Conclusion:
Independent action of these anti-oxidants was modest but non-significant, whereas in combination the action was good indicating synergistic action of these molecules. It is possible that the synergistic effect due to bio-energetic and ATP (adenosine tri phosphotase) producing activity of coenzyme Q10. The activity of coenzyme Q10 is potentiated by the activity of Vitamin E and Vitamin C. Coenzyme Q10 also has a sparing effect on intracellular levels of Vitamin E since Q 10 has multiple beneficiary activities it should have decreased the anti-oxidants conjunction by the cells of the body and potentiated the overall composition.

We Claim:
1. A process of preparing a new antioxidant composition for the treatment
of chronic renal failure and heart failure comprising mixing the following
anti-oxidants in the proportions indicated there against:
Coenzyme Q10 (Ubiquinone) - 10-40% by wt.
Vitamin E - 30-45% by wt.
Vitamin C - 30-45% by wt.
and the balance if any adding a conventional filler fof preparing the tablet or capsule.
2. A process as claimed in claim 1 wherein said filler used is Psylliul.
3. A process as claimed in claim 1 wherein the proportions of the ingredients in the anti-oxidant composition are as follows
Coenzyme Q10 - 20% by wt.
Vitamin E - 40%bywt.
Vitamin C - 40% by wt.
4. A process as claimed in claim 1 wherein the proportions of the
ingredients in the anti-oxidant composition are as follows
Coenzyme Q10 - 30%bywt.
Vitamin E - 30% by wt.
Vitamin C - 30% by wt.
Filler - 10%bywt.
5. A process of preparing a new anti-oxidant composition for the treatment
of chronic renal failure and heart failure substantially as herein described
with reference to the foregoing examples.

Dated this 12th day of July, 2000
of Anand & Anand, Advocates
Agents for the Applicants

Documents:

668-mum-2000-abstract(17-07-2000).doc

668-mum-2000-abstract(17-07-2000).pdf

668-mum-2000-cancelled pages(13-11-2000).pdf

668-mum-2000-claims(granted)-(12-11-2000).doc

668-mum-2000-claims(granted)-(12-11-2000).pdf

668-mum-2000-correspondence(16-11-2000).pdf

668-mum-2000-correspondence(ipo)-(14-06-2007).pdf

668-mum-2000-form 1(17-7-2000).pdf

668-mum-2000-form 3(17-7-2000).pdf

668-mum-2000-form 3a(2)(granted)-(12-11-2000).doc

668-mum-2000-form 3a(2)(granted)-(12-11-2000).pdf

668-mum-2000-other document(17-7-2000).pdf

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Patent Number

207585

Indian Patent Application Number

668/MUM/2000

PG Journal Number

43/2008

Publication Date

24-Oct-2008

Grant Date

14-Jun-2007

Date of Filing

17-Jul-2000

Name of Patentee

PROF. RAM BAHADUR SINGH

Applicant Address

SAFE ENTERPRISES, 6 CITIZEN APARTMENT, 30TH ROAD, BANDRA, MUMBAI - 400 050, MAHARASHTRA, INDIA.

Inventors:

#	Inventor's Name	Inventor's Address
1	PROF. RAM BAHADUR SINGH	SAFE ENTERPRISES, 6 CITIZEN APARTMENT, 30th ROAD, BANDRA, MUMBAI - 400 050, MAHARASHTRA, INDIA.
2	DR. MOHAMMAD ARIF NIAZ	SAFE ENTERPRISES, 6 CITIZEN APARTMENT, 30th ROAD, BANDRA, MUMBAI - 400 050, MAHARASHTRA, INDIA.

PCT International Classification Number

C09K 15/00

PCT International Application Number

N/A

PCT International Filing date

PCT Conventions:

#	PCT Application Number	Date of Convention	Priority Country
1			NA