ENZYME COMPOSITION FOR MANAGEMENT OF METABOLIC HEALTH

Abstract

The present invention discloses compositions containing digestive enzymes for the management and maintenance of metabolic health. Specifically, the invention discloses the use of digestive enzymes comprising -amylase, lactase, lipase, cellulase and Neutral protease or Acid protease in weight management, reducing cholesterol levels, maintaining healthy gut and improving quality of life.

Claims

1. A method for management and improvement of metabolic health in mammals, said method comprising step of administering effective concentration of enzyme blend comprising of -amylase, cellulase, lipase, lactase and neutral protease or acid protease to mammals in need of such therapy.

2. The method as in claim 1, wherein the improvement in metabolic health is brought about by a) reducing and/or maintaining constant body weight and BMI, b) reducing lipid levels, and c) improving gut health and quality of life.

3. The method as in claim 1, wherein the effective concentration of the enzyme blend is 20-200 mg, with concentrations of the individual enzymes being a) -amylase: not less than 24000 DU/g, b) cellulase: not less than 1100 CU/g, c) lipase: not less than 200 FIP/g, d) lactase: not less than 4000 ALU/g and e) neutral or acid protease: not less than 6000 PC/g.

4. The method as in claim 1, wherein the composition is formulated with pharmaceutically/nutraceutically acceptable excipients, adjuvants, diluents or carriers and administered orally as a stand-alone or in combination with standard treatment, in the form of tablets, capsules, syrups, gummies, powders, suspensions, emulsions, chewables, candies and eatables.

5. The method as in claim 1, wherein the standard treatment involves administration of one or more drugs selected from the group consisting of antibiotics, statins, cholesterol absorption inhibitors, Insulin sensitizing agents, GLP-1 receptor agonists and DPP-4 inhibitors.

6. The method as in claim 1, wherein the mammal is preferably human.

7. A method for reducing hyperlipidemia in mammals, said method comprising step of administering effective concentration of enzyme blend comprising of -amylase, cellulase, lipase, lactase and neutral protease or acid protease to said mammals to bring about the effect of reduction in the levels of a) total cholesterol, b) LDL and LDL-Cholesterol, and c) cholesterol:HDL ratio.

8. The method as in claim 7, wherein the effective concentration of the enzyme blend is 20-200 mg, with concentrations of the individual enzymes being a) -amylase: not less than 24000 DU/g, b) cellulase: not less than 1100 CU/g, c) lipase: not less than 200 FIP/g, d) lactase: not less than 4000 ALU/g and e) neutral or acid protease: not less than 6000 PC/g.

9. The method as in claim 7, wherein the composition is formulated with pharmaceutically/nutraceutically acceptable excipients, adjuvants, diluents or carriers and administered orally as a stand-alone or in combination with standard treatment, in the form of tablets, capsules, syrups, gummies, powders, suspensions, emulsions, chewables, candies and eatables.

10. The method as in claim 7, wherein the standard treatment involves administration of one or more drugs selected from the group consisting of antibiotics, statins, cholesterol absorption inhibitors, Insulin sensitizing agents, GLP-1 receptor agonists and DPP-4 inhibitors.

11. The method as in claim 7, wherein the mammal is preferably human.

12. A method for the improvement in gut health in mammals, said method comprising steps of administering effective amounts of enzyme blend comprising of -amylase, cellulase, lipase, lactase and neutral protease or acid protease to mammals showing symptoms of gastrointestinal discomfort.

13. The method as in claim 12, wherein the symptoms associated with gastrointestinal discomfort include presence of E. coli in stool, gastric reflux, abdominal pain, indigestion, diarrhoea and constipation.

14. The method as in claim 12, wherein the effective concentration of the enzyme blend is 20-200 mg, with the concentration of the individual enzymes being a) -amylase: not less than 24000 DU/g, b) cellulase: not less than 1100 CU/g, c) lipase: not less than 200 FIP/g, d) lactase: not less than 4000 ALU/g and e) neutral or acid protease: not less than 6000 PC/g.

15. The method as in claim 12, wherein the composition is formulated with pharmaceutically/nutraceutically acceptable excipients, adjuvants, diluents or carriers and administered orally as a stand-alone or in combination with standard treatment, in the form of tablets, capsules, syrups, gummies, powders, suspensions, emulsions, chewables, candies and eatables.

16. The method as in claim 12, wherein the standard treatment involves administration of one or more drugs selected from the group consisting of antibiotics, statins, cholesterol absorption inhibitors, Insulin sensitizing agents, GLP-1 receptor agonists and DPP-4 inhibitors.

17. The method as in claim 12, wherein the mammal is preferably human.

Description

EXAMPLE 1

Methods

[0017] Study Objectives:

[0018] The objective of the study is to assess the efficacy of enzyme blend in the management of metabolic health. The primary objectives include evaluating the change in the body weight and the body mass index (BMI) by the enzyme blend and changes in biochemical parameters (Serum lipids, Stool Analysis). The changes in the gut health will be evaluated using a questionnaire. The study also assesses the quality of life as a secondary objective.

[0019] Selection of Subjects

Number of subjects: 30

Nature of Population

[0020] Male and female healthy subjects between the age group of 18-65 years with BMI between 30 to 34.9 kg/m.sup.2 were screened for this study. The screening procedures included medical history, medication history, treatment history, co-morbid conditions, physical examination, vital signs and urine pregnancy test for women, lipid profile, hematology, biochemistry and stool analysis. Subjects who met the inclusion criteria or for whom the abnormalities in laboratory parameters are not clinically significant were selected for the study.

[0021] Inclusion Criteria

[0022] Subject who met the below mentioned inclusion criteria were enrolled in the study

Male and/or female.
Age between 18 to 65 years.
BMI between 30 to 34.9 Kg/m.sup.2
Willing to come for regular followup visits.
Able to give written informed consent.

[0023] Exclusion Criteria

[0024] Subject who met any of the below mentioned exclusion criteria were excluded from the study.

Intake of over the counter weight loss agents, centrally acting appetite suppressants in the previous six months.
Pathophysiologic/genetic syndromes associated with obesity (Cushing's syndrome, Turner's syndrome, Praderwilli syndrome).
Patients on prolonged (>6weeks) medication with corticosteroids, antidepressants, anticholinergics, etc. or any other drugs that may have an influence on the outcome of the study.
Patients suffering from major systemic illness necessitating long term drug treatment (Rheumatoid arthritis, Tuberculosis, Psycho-Neuro-Endocrinal disorders, etc.)
Symptomatic patient with clinical evidence of Heart failure.
History of HIV and other viral infections.
Alcoholics and/or drug abusers.
Prior surgical therapy for obesity
H/o hypersensitivity to any of the trial drugs or their ingredients.
Pregnant/lactating, woman.

[0025] Study Design

[0026] Subjects were screened only after obtaining their informed consent. Participants who met the inclusion criteria and did not violate the exclusion criteria were administered with a capsule containing the enzyme blend(DigeZymeSabinsa Corporation, USA) for a period of 90 days. Table 1 provides the constituents on the enzyme blend

TABLE-US-00001 TABLE 1 Active ingredient of Capsule S. No. Enzyme Source 1. Alpha-amylase Aspergillus oryzae 2. Lactase Aspergillus oryzae 3. Lipase Rhizopus oryzae 4. Cellulase Trichoderma longibrachiatum, Trichoderma reesei 5. Neutral Protease Bacillus subtilis Acid Protease Aspergillus oryzae

[0027] Screening Visit

[0028] A total of 30 participants were enrolled after obtaining informed consent, medical & medication history, treatment history, co-morbid conditions, physical examination, demographic data, vital signs, blood sample for laboratory analysis like haematology, lipid profile & biochemistry, urine sample for urine pregnancy test & stool analysis (routine examination , culture count & sensitivity for E. Coli & Bifidogenic bacteria) & Anthropometric measurements (the size, weight, and proportions of the body hips & waist) were performed and recorded.

[0029] Day 0 (Visit 1)

[0030] Evaluations for Inclusion & exclusion criteria were carried out and eligible subjects were enrolled into the study. Subjects were instructed on their daily dose of study supplement. Subjects were provided with study visit plan. Vital signs & anthropometric measurements were documented in the respective CRFs. Concomitant medications, if any, were recorded.

Study supplements were provided to all subjects as per the randomization code, subjects were instructed to take daily the enzyme blend capsules on days not coming to the clinic. Assessments using subjective questionnaires were performed.

[0031] Day 45 (Visit 2)

[0032] Study supplements were provided to all subjects as per the randomization code, subjects were instructed to take daily the enzyme blend supplement capsules on days not corning to the clinic. Demographics, Physical examination, vital signs & anthropometric measurements & lipid profile were documented in the respective CRFs. Photographs of the subjects were taken. Adverse Events and concomitant medications, if any, were recorded. Assessments using subjective questionnaires were performed.

[0033] Day 90 (Final Visit)

[0034] Demographics, Physical examination, vital signs & anthropometric measurements were documented in the respective CRFs. Urine pregnancy test for women were performed. Stool analysis, haematology; serum biochemistry and lipid profile tests were performed. Adverse Events and concomitant medications, if any, were recorded by the site personnel. Concomitant medication, if any, was recorded. Photographs of the subjects were taken. Assessments using subjective questionnaires were performed.

[0035] Follow Up Visit

Telephonic Follow up (15 days from the final visit for the overall general well being of the subject)

[0036] Efficacy and Safety Variables

Efficacy Evaluation

[0037] Efficacy was analysed by

[0038] Changes in the body weight and the body mass index.

[0039] Changes in the lab investigations (Serum lipids, Stool Analysis).

[0040] Changes in the gut health questionnaire

[0041] Improvement in Quality of Life as per Quality of Life Questionnaire

[0042] All the subjects enrolled in the study were evaluated for the effect of study supplement in healthy volunteers for metabolic health during the end of study or at the time of discontinuation of the treatment in comparison with baseline.

[0043] Safety Data

[0044] Safety measurements included monitoring of AEs (including worsening of gastrointestinal symptoms), physical examination results and vital signs were performed

[0045] Statistical Methods

[0046] The primary efficacy measure was the decrease in the body weight and BMI, changes in the lab investigations (serum lipids & stool analysis), changes in the gut health questionnaire from baseline to end of the study. Secondary efficacy measures included Improvement in Quality of Life as per Quality of Life Questionnaire, tolerability of the study drug by assessing for the adverse events and other physical signs or symptoms by the Investigator. For statistical analysis purposes the baseline values was compared to that of all visit values by statistical analysis in terms of mean, standard deviation. The results between the first and the subsequent visiting were analyses using paired t test.

EXAMPLE 2

Results

[0047] Baseline and Demographic Characteristic

[0048] Thirty healthy subjects with BMI between 30 to 34.9 Kg/m.sup.2 who were deemed eligible to have met the study requirements were administered the digestive enzyme blend for a period of 90 days. The demographic data and baseline characteristics of the enrolled patient population are summarized in Table 2.

TABLE-US-00002 TABLE 2 Baseline and demographic characteristic Enzyme Parameter Statistics blend (N = 30) Age (years) Mean SD 36 9.77 Median 35 Min; Max 22, 59 Gender, n (%) Male 16 (53.33) Female 14 (46.67) Body weight (kg) Mean SD 80 9.62 Median 82 Min; Max 64, 98 Height (cm) Mean SD 159 9.95 Median 162 Min; Max 143, 175

[0049] Efficacy Evaluation

[0050] Decrease in the Body Weight and the Body Mass Index (BMI)

[0051] The changes in the body weight and the body mass index (BMI) are presented in Table 3.

TABLE-US-00003 TABLE 3 Changes in bodyweight and BMI Body weight (kg) BMI (kg/m.sup.2) Baseline (Day 0) 80.12 9.62 31.77 1.07 Visit 2 (Day 45) 79.55 9.59* 31.55 1.11* Visit 3 (Day 90) 78.67 9.67* 31.17 1.73* *Statistically significant (P < 0.05) compared to baseline (Paired t-test)

[0052] The results indicated that the digestive enzyme supplement showed a slight decrease in bodyweight and BMI. The enzyme blend also helps in maintaining constant body weight with no signs of increase in BMI.

[0053] Changes in the Lab Investigations

[0054] The lipid profile was estimated at the baseline and during each visit. The results are tabulated in table 4.

TABLE-US-00004 TABLE 4 Lipid profile in subjects supplemented with enzyme blend Baseline Visit 2 Visit 3 (Day 0) (Day 45) (Day 90) Total cholesterol 188.97 32.11 182.67 28.45* 176.03 26.73* (mg/dl) Triglycerides 157.47 57.59 169.27 57.43 157.53 48.88 (mg/dl) HDL (mg/dl) 38.5 8.45 41.57 9.58 37.23 7.33 LDL (mg/dl) 129.27 26.32 120.47 25.95* 113.53 25.05* VLDL (mg/dl) 21.2 10.49 21.57 8.30 27.37 20.45 Cholesterol:HDL 5.08 1.20 4.57 1.05* 4.79 1.27* ratio *Statistically significant (P < 0.05) compared to baseline (Paired t-test)

[0055] The results indicated that the enzyme blend reduced the total cholesterol. LDL levels and cholesterol:HDL ratio

[0056] Stool Analysis

[0057] In many Clinical studies the reduced numbers of Bifidobacterium and Bacteroides and increased numbers of Staphylococcus, Enterobacteriaceae and Escherichia coli were detected in overweight compared with normal-weight population. E. coli numbers are reported to be higher in population with excessive weight gain than in population with normal weight. The present study analysed the E. coli numbers in stool and its relationship with body weight and weight gain. The E. coli numbers in stools was tabulated in Table 5.

TABLE-US-00005 TABLE 5 Presence of E. coli in stool E. coli cells (No.) Baseline (Day 0) 70.83 15.10 Visit 3 (Day 90) 63.08 14.54* *Statistically significant (P < 0.05) compared to baseline (Paired t-test)

[0058] E. coli numbers was found to be lower in visit 3 indicating a change in the gut microbiota as a result of enzyme supplementation. This reduction correlated with the change in body weight in BMI indicating that the change in gut microbiota reduces bodyweight and helps in the management of metabolic health.

The change in gut microbiota also resulted in the improvement of gut health, which is indicated below:

[0059] Gut Health Assessment

[0060] The gut health questionnaire helps in determining the imbalance in the digestive system. In the present study, it was assessed in baseline, visit 3 and final visit and the result are tabulated in table 6.

TABLE-US-00006 TABLE 6 Gut health assessment score Gut health assessment Score Baseline (Day 0) 26.57 11.35 Visit 2 (Day 45) 20.13 6.45* Visit 3 (Day 90) 14.33 4.78* *Statistically significant (P < 0.05) compared to baseline (Paired t-test)

[0061] In the baseline visit the mean of Gut Health Assessment score was found to be 26.57+11.35 and significantly reduced to 20.136.45 in visit 2 and 14.334.78 in final visit indicating that the enzyme supplement significantly improved the gut health.

[0062] Gastrointestinal Symptom Rating Scale (GSRS)

[0063] The GSRS is a disease-specific instrument of 15 items combined into five symptom clusters depicting reflux, abdominal pain, indigestion, diarrhea and constipation. The GSRS has a four-point graded scale where 0 represents absence of troublesome symptoms and 3 represent very troublesome symptoms. In the current investigation, the GSRS was in baseline, visit 2 and final visit. The results are indicated in table 7.

TABLE-US-00007 TABLE 7 Change in Gastrointestinal Symptom Rating Scale Gut health assessment Score Baseline (Day 0) 6.9 2.59 Visit 2 (Day 45) 5.13 1.78* Visit 3 (Day 90) 3.57 1.48* *Statistically significant (P < 0.05) compared to baseline (Paired t-test)

[0064] The results indicated a significant reduction in GSRS score in the follow up visits when compared to the initial visit indicating that the enzyme blend supplement improved the gastrointestinal function.

[0065] Improvement in Quality of Life

[0066] Physically, sonic of the problems associated with obesity are hypertension, coronary arteriosclerosis, elevated cholesterol, type 2 diabetes, joint problems, stroke, and certain types of cancers. Psychologically, obesity is associated with a myriad of problems including lower self-concept, negative self evaluation, and decreased self-image. Socially, obese individuals often encounter discrimination and prejudice, which further perpetuate negative economic and social consequences. In general, obesity is associated with decrements in overall quality of life whether it is physical, psychological, or social. The lower the score, the more quality of life was thought to be impaired. The impact of being overweight and obese was studied on Baseline, Visit 2 and Visit 3 using a questionnaire. The results are presented in Table 8.

TABLE-US-00008 TABLE 8 Mean change in Quality of Life Quality of life score Baseline (Day 0) 30.93 3.42 Visit 2 (Day 45) 31.4 3.09 Visit 3 (Day 90) 35.23 2.82* *Statistically significant (P < 0.05) compared to baseline (Paired t-test)
The results indicated that the enzyme blend significantly improved the quality of life.

[0067] Overall, supplementation of the enzyme blend containing -amylase, protease, lipase, cellulose, and lactase, was effective in decreasing/maintaining body weight and BMI, improving gut health and gastrointestinal efficiency by reducing E. coli and alleviating the symptoms of gut disorders. The study indicated that the enzyme blend can be effectively used as a supplement for the management and maintenance of metabolic health. Since, dietary supplements also improve the efficacy of the treatment drugs, the enzyme blend can also be supplemented with the standard drugs used for the treatment of metabolic syndrome.

EXAMPLE 3

Compositions/Formulations Containing Digestive Enzymes

[0068] Tables 9-13 provide illustrative examples of formulations containing digestive enzyme blend. The digestive enzyme blend disclosed in the current application is available in the name DigeZyme, from Sabina Corporation, USA.

TABLE-US-00009 TABLE 9 DigeZyme Capsule Active Ingredients DigeZyme 20-200 mg containing -amylase 24,000 DU/g, lactase 4000 LAC/g, cellulase 1100 CU/g, lipase 200 FIP/g, acid protease 10,000 HUT/g Excipients Maltodextrin, Magnesium stearate

TABLE-US-00010 TABLE 10 DigeZyme Capsule Active Ingredients DigeZyme 20-200 mg containing -amylase 24,000 DU/g, lactase 4000 LAC/g, cellulase 1100 CU/g, lipase 200 FIP/g, neutral protease 6000 PC/g Excipients Maltodextrin, Magnesium stearate

TABLE-US-00011 TABLE 11 DigeZyme Capsule Active Ingredients DigeZyme 20-200 mg containing -amylase 24,000 DU/g, lactase 4000 LAC/g, cellulase 1100 CU/g, lipase 200 FIP/g, neutral protease 7000 PC/g Excipients Maltodextrin, Magnesium stearate

TABLE-US-00012 TABLE 12 DigeZyme Capsule Active Ingredients DigeZyme 20-200 mg containing -amylase 24,000 DU/g, lactase 30,000 LAC/g, cellulose 1100 CU/g, lipase 200 FIP/g, neutral protease 7000 PC/g Excipients Maltodextrin, Magnesium stearate

TABLE-US-00013 TABLE 12 DigeZyme Drink Mix Active Ingredients DigeZyme (-amylase, lactase, lipase, cellulase, Neutral protease or Acid protease) - 20 to 200 mg Beet root extract, Taurine, Vitamin B6, Vitamin B 12 Excipients Maltodextrin, Citric acid, Sucralose, Flavouring agents

[0069] The above formulations are merely illustrative examples, any formulation containing the above active ingredient intended for the said purpose will be considered equivalent.

[0070] Other modifications and variations to the invention will be apparent to those skilled in the art from the foregoing disclosure and teachings. Thus, while only certain embodiments of the invention have been specifically described herein, it will be apparent that numerous modifications may be made thereto without departing from the spirit and scope of the invention.