Title of Invention

PREPARATION OF DIGESTIVE ENZYMES CONTAINING DRY SYRUP

Abstract This invention provides a process for preparation of Dry Syrup containing digestive enzymes as free flowing granules having sufficient shelf life, comprising of (i) a pharmaceutically active digestive enzymes, (ii) water soluble sweetener, (iii) suspending agent, and (iv) stabilizer; which exhibits good stability and reconstitution into palatable syrup used for digestive disorders.
Full Text FORM 2
THE PATENT ACT 1970
(39 of 1970)
&
The Patents Rules, 2003
COMPLETE SPECIFICATION
(See section 10 and rulel3)
1. TITLE OF THE INVENTION:
Preparation of Digestive Enzymes containing Dry Syrup
2. APPLICANT (S)
(a) NAME:
(b)NATIONALITY:
(c) ADDRESS:
3. PREAMBLE TO THE DESCRIPTION
The following specification particularly describes the invention and the manner in
which it is to be performed.

Technical field of the Invention:
This invention provides a process for preparation of Dry Syrup containing digestive enzymes as free flowing granules having sufficient shelf life.
Background of the Invention:
Enzymes are proteins whose biological function is the catalysis of chemical reactions in living systems. Enzymes are composed of two parts, a protein portion called as apoenzyme and a nonprotein portion, either a coenzyme (organic) or cofactor (inorganic). Enzymes combine with the substances on which they act called as Substrate to form intermediate enzyme-Substrate complex, which is then converted to a reaction product, and liberated enzyme, which continues its catalytic function. Enzymes are highly specific; a few exhibit absolute specificity and catalyze only one particular reaction, while others are specific for a particular type of chemical bond, functional group or Stereo isomeric structure.
The Commission of Enzymes (EC) of the International Union of Biochemistry has classified the enzymes into six general groups:
1) Hydrolyses,
2) Isomerases,
3) Ligases,
4) Lyases,
5) Oxidoreductases
6) Transferases
The enzymes involved in food decomposition and in the digestive process are hydrolases. When raw food is ingested, enzymes present within the food are released, thereby assisting the body's digestive processes in breaking down the food into its simplest components for utilization within the body.
There are several categories of food enzymes such as:
1) Lipase which breaks down fats into free fatty acids and glycerol
2) Protease which breaks down long protein chain into smaller amino acid chains
2

3) Amylase, which reduces large carbohydrates to disaccharide including sucrose, lactose and maltose.
4) Lactase which breaks down lactose
Several factors are known to contribute to the development of digestive problems. Poor eating habits, such as inadequate chewing of food, "eating on the run," or eating late in the day, can result in inadequate enzyme production. Certain dietary choices, such as excessive consumption of alcohol, refined carbohydrates and fat, as well as a high meat and cooked food diets with few raw, enzyme-rich foods may also place stress on the body's digestive capacity. In addition, problems with digestion can occur simply as a result of aging. It is fairly common for elderly individuals to experience both a decrease in hydrochloric acid production as well as a general decline in digestive enzyme secretion. With an increasing aging population burdened with unhealthful diets and stressful life styles, it is likely that healthcare professionals will see more and more patients developing digestive problems. Currently, it is estimated that fifty-eight percent of the population suffers from some type of digestive disorder. A lack of optimal digestive function associated with enzyme inadequacy may lead to malabsorption and a host of related conditions. Consequences of malabsorption can include impaired immunity, allergic reaction, poor wound healing, skin problems and mood swings. Supplemental enzymes can improve the level of digestion and help assure that the maximum level of nutrient absorption is attained.
Perhaps one largely overlooked factor that may contribute to problems with digestion may be the lack of raw, uncooked foods present in the average diet. Raw foods contain many enzymes that may help facilitate digestion. When they are eaten, mastication breaks down cell membranes allowing the release and activation of the naturally occurring enzymes present in the food. These enzymes may play a beneficial role by initiating the breakdown of the food and giving the body a head start on the digestive process. Unfortunately, in this part of the world we tend to eat mostly cooked foods. When a food is cooked at 118°F or greater the natural enzymes are destroyed. Even though cooking destroys these helpful, naturally occurring enzymes, it may not be reasonable to suggest that a total raw foods diet is appropriate. For food safety reasons alone, this practice would not be advisable. Furthermore, some foods such as carrots and broccoli provide adequate nutrient value when cooked and may
3

actually cause fewer gastrointestinal symptoms in patients who have difficulty in digesting cellulose. Inclusion of appropriate raw foods combined with enzyme replacement may be the most balanced and rational way to support those with digestive enzyme insufficiencies.
In addition to consuming adequate levels of raw foods, a common approach to supporting the patient with digestive enzyme inadequacy is oral enzyme replacement. This entails providing enzyme supplements, whether they be animal or nonanimal derived, in the quantity necessary to maintain adequate digestive capacity and facilitate absorption of essential nutrients. Oral enzyme replacement has proven to be helpful over many years of use. In addition to the well known benefits associated with enzyme replacement, it has been suggested that oral supplementation with enzymes may have a sparing effect on the body's own digestive enzymes.
Supplemental enzymes of microbial and plant origin work at the pH found in the upper stomach. Food sits in the upper portion of the stomach for as long as an hour before gastric secretions begin their action. Several studies have shown that the enzymes in saliva continue their digestive activity in the upper stomach and can digest up to 30% of the ingested protein, 60% of ingested starch and 10% of ingested fat during the 30 to 60 minutes after consumption. Although salivary enzymes accomplish a significant amount of digestion, their activity is limited to a pH level above 5.0. Supplemental microbial enzymes, and some plant enzymes, are active in the pH range of 3.0 to 9.0 and can facilitate the hydrolysis of a much larger amount of protein, carbohydrates and fat before Hydrochloric Acid is secreted in sufficient amounts to neutralize their activity. Obviously, these enzymes can contribute significantly in improving food nutrient utilization.
The stabilization of enzymatic activity is a standing problem in all areas of technology where enzymes are likely to be used. Stability in this sense stands for resistance to decrease in enzymatic activity prior to usage e.g. under storage conditions. Enzyme stability problems are most important when the enzyme containing composition or additive formulated with water or is used in aqueous solutions.
4

In aqueous condition the enzyme preparations denatures due to extremes of pH or temperature upon storage. So proposed dry syrup is solid dosage form where enzymes in solid state remained stable for long time and also stabilizers are added to stabilize the enzymes on storage condition.
References Cited
Diehl, et al.; United States Patent 4011169, March 1997.
Bilton, et al.; United States Patent 4447412, May 8, 1984.
Vidal, et al.; United States Patent 4116772, September 26, 1978.
Liliana N. Ceci, et al.; "Effect of pH, Heat and Calcium ions on amylase", Food
Technol. Biotechnol. 40,33-38(2002)
Indian Pharmacopoeia, 1996.
Remington's Pharmaceutical Sciences, 18th edition, 1990.
Schlenker ED. Nutrition In Aging. 2nd ed. St. Louis, MO: Mosby, 1993;88-91.
The Trends Journal, Summer Issue, 1994.
Howell E. Enzyme Nutrition-The Food Enzyme Concept. Wayne, NJ: Avery
Publishing Group 1985; 29.
Alternative Medicine, the Definitive Guide. Future Medicine Publishing: Puyallup,
WA. 1993; 215-22.
Mangels AR, et al. The bioavailability to humans of ascorbic acid from oranges,
orange juice and cooked broccoli is similar to that of synthetic ascorbic acid. J Nutr
1993; 123:1054-101.
Warren KW, et al. Life after total pancreatectomy for chronic pancreatitis. Ann. Surg
1966; 164:830-34.
Griffin SM, et al. Acid resistant lipase as replacement therapy in chronic pancreatic
exocrine insufficiency: a study in dogs. Gut 1989; 30:1012-15.
Schneider MU, et al. Pancreatic enzyme replacement therapy: comparative effects of
conventional and enteric-coated microspheric pancreatin and acid-stable fungal
enzyme preparations on steatorrhea in chronic pancreatitis. Hepato-gastroenterol
1985; 32:97-102.
DiMagno EP, et al. Comparative effects of antacids, cimetidine and enteric coating on
the therapeutic response to oral enzymes in severe pancreatic insufficiency. NEJM
1977; 297(16): 854-58.
5

Graham DY. Enzyme replacement therapy of exocrine pancreatic insufficiency in
man.NEJM 1977;296:1314-17.
Liebow C, Rothman SS. Enteropancreatic circulation of digestive enzymes, Science
1975; 189:472-74.
Murray M, Pizzorno J. Encyclopedia of Natural Medicine, (Prima Publishing,
Rocklin, CA. 522-23, 1991).
Prochaska LJ and Piekutowski WV "On the synergistic effects of enzymes in food
with enzymes in the human body. A literature survey and analytical report." Medical
Hypotheses 42: 355-62 (1994).
Schwimmer S. Source Book of Food Enzymology. (Westport, CT: The AVI
Publishing Company, Inc., 1981.
Whitney EN, Cataldo CB, and Rolfes SR. Understanding Normal and Clinical
Nutrition.
(St. Paul, New York, Los Angeles, San Francisco: West Publishing Company, 1991.)
Summary of the Invention
This invention provides a process for preparation of Dry Syrup containing digestive enzymes as free flowing granules having sufficient shelf life.
Digestives enzymes in dry powder form are stable for upto 2-3 years if stored at controlled room temperature, but they are quite unstable in aqueous condition even at controlled room temperature. Many available digestive enzyme preparations are in liquid dosage forms, which are not stable upon storage. Therefore it is the object of the present invention to provide the stable formulation for digestive disorders. In the present invention, formulation developed is dry syrup, which is free flowing granular form, and it is to be reconstituted when required. It contains digestive enzymes, preservative, water soluble sweetener, suspending agent, glidant, stabilizer, colour and flavor. The excipients used are relatively inexpensive and easily commercially available. Also the process adopted is simple and cost effective.
Detailed description of the Invention:
Dry syrup is defined as prepared solid mixture intended for reconstitution just before use with appropriate vehicle. The reconstituted syrup is the formulation of choice
6

when drug stability is major concern and this dosage form is formulated to avoid the physical stability problems often encountered in conventional syrups. Also formulation for reconstitution reduces the weight of the final product because the aqueous vehicle is absent and, consequently, transportation expenses may be reduced. The dry syrup may be shipped without regard to seasonal temperatures because its physical stability is less susceptible to temperature extremes as compared with conventional syrups.
Preferred digestive enzymes in the context of the invention are: amylase, protease, peptidase, lipase, lactase, cellulase, maltase, invertase, pepsin, bromelain, papain, chymotrypsin, trypsin, Pancreatine, glucoamylase, maltdistase, sucrase. The formulation also include at least one water soluble sweetner selected from the group consisting of sucrose, dextrose, mannitol, Sorbitol, xylitol in 50-95 weight parts per 100 weight parts of the granules and at least one suspending agent from the group of natural gums consisting of acacia, guar gum, tragacanth, Xanthan gum in 0.5 to 1.5 weight parts per 100 weight parts of the granules.
The formulation prepared by wet granulation method include at least one water soluble binder from the group consisting of sucrose syrup, povidone, Hydroxy propyl cellulose, Hydroxy propyl methyl cellulose, hydroxy ethyl cellulose, Sodium Carboxy methyl cellulose in 0.5 to 5 weight parts per 100 weight parts of the granules and solvent used from the group consisting of water, Iso propyl alcohol.
Stabiliser used includes at least one from the group consisting of calcium chloride, calcium Gluconate, calcium lactate, and calcium propionate in 0.01 to 1 weight parts per 100 weight parts of the granules. The formulation also include at least one glidant from the group consisting of colloidal silicon dioxide, starch, talc, magnesium trisilicate in 0.1 to 1 weight parts per 100 weight parts of the granules and also include suitable FDA approved preservative, water soluble colourants and powder flavours.
Digestive enzymes used in present invention are fungal alpha amylase and pepsin where label claim of formulation is Each 5 ml of reconstituted syrup contains Fungal alpha amylase derived from Aspergillus oryzae ( 1:1200) 50 mg and Pepsin (1:3000) 10 mg. Appropriate overages of enzymes added to compensate the losses on storage.
7

These digestive enzymes formulated by two method, wet granulation and dry granulation.
In wet granulation method, digestive enzymes are mixed with sweetener and sugar syrup is used as binder and wet mass is dried at 40-45°C to form dry granules and remainder excipients are added after drying. In case of dry granulation method, digestive enzymes and other excipients are mixed in dry form. Here stabilizer used is Calcium chloride, which act as cofactor and stabilizes the enzymes. (Ref: "Effect of pH, Heat and Calcium ions on amylase"; Liliana N. Ceci, Jorge E.Lozano, Food Technol. Biotechnol. 40(2002) 33-38)
The invention will be further described by reference to the following detailed examples wherein fungal alpha amylase and Pepsin was obtained from Biocon India; other excipients used are of Pharmacopoeial grades.
The concentration of Fungal Alpha amylase and pepsin was estimated by method given by Indian Pharmacopoeia where the method estimates the substrate, remaining after the assay time interval for enzyme hydrolysis under defined conditions to evaluate the enzymatic activity.
Example 1: Granules preparation by wet granulation technique
90 gm of fungal alpha amylase and 18 gm of Pepsin passed through 40 # sieve and mixed homogenously. Sifted 1.362 kg of Pharma grade sucrose through 40 # sieve and blend of enzymes mixed with sucrose in geometric fashion.
In 120 ml purified water dissolved 2.4 gm of sodium benzoate, 2.4 gm of Calcium chloride and 80 gm Sucrose by heating at 90 - 100°C, then solution cooled down to 37°C and to it added 0.5 gm Quinoline Yellow Colour and stirred to dissolve Colour completely. This solution added to above powder blend and mixed to form coherent mass. Then this mass is passed through 8 # sieve and dried at 40 - 45 °C for 2 hours. The dried granules passed through 18 # sieve.
8

10 gm of Guar gum, 5 gm of Colloidal Silicon Dioxide and 30 gm of Pineapple powder flavour passed through 40 # and added to dried granules and mix for 5 mins in cone blender. Light yellow, pineapple flavoured free flowing granules obtained with moisture content 0.45 %.
Long term stability carried out at 30°C±2°C / 65 % RH ± 5 % RH and results are shown in Table I.
Table I:

Time in months % Retention of Fungal Alpha amylase % Retention of Pepsin
0 141.23% 140.39 %
3 134.30% 135.42%
6 129.18% 130.29%
9 121.67% 123.53 %
12 116.92% 120.19%
18 105.35 % 104.08 %
24 93.80 % 91.98%
Example 2: Granules preparation by dry granulation technique
14.37 kg of Pharma grade sucrose and 50 gm of Colloidal Silicon Dioxide passed through 40 # sieve and mixed in cone blender for 15 minutes.
900 gm of fungal alpha amylase and 180 gm of Pepsin passed through 40 # sieve and mixed homogenously. Then blend of enzymes mixed with sucrose in geometric fashion.
Triturated 5 gm Sunset Yellow Colour with 150 gm of Guar gum and then passed through 60 #. 0.3 kg Orange powder flavor, 0.024 kg of sodium Benzoate and 0.024 kg of Calcium Chloride passed through 40 #. All bends are mixed in cone blender for
9

20 minutes. Light orange coloured, orange flavoured free flowing granules obtained with moisture content 0.28 %.
Long term stability carried out at 30°C±2°C / 65 % RH ± 5 % RH and results are shown in following Table II.
Table II:

Time in months % Retention of Fungal Alpha amylase % Retention of Pepsin
0 148.00 % 144.29 %
3 141.69% 138.21 %
6 135.88% 132.47 %
9 129.22% 126.04 %
12 123.50% 120.14%
18 110.29% 108.57%
24 98.08 % 96.12%
The stability of this preparation was considered to be excellent which is stable over 2 years.
Example II granules shown faster reconstitution and better palatability than Example I.
10

16 gm of Example 2 granules reconstituted to 60 ml with boiled and cooled drinking water and kept for long term study at 30°C±2°C / 65 % RH ± 5 % RH and results are shown in Table III.
Table III

Time in days % Retention of Fungal Alpha amylase % Retention of Pepsin
0 148.00 % 144.29 %
5 130.23 % 135.87%
10 120.91 % 127.33 %
15 105.78% 110.11 %
20 93.56 % 105.45 %
The reconstituted syrup is stable upto 15 days.
Many variations and changes may be made while remaining within scope of the Invention.
11

We Claim,
1. This invention provides a process for preparation of Stable Dry Syrup containing
digestive enzymes as free flowing granules comprising of (i) a pharmaceutically active digestive enzymes, (ii) water soluble sweetener, (iii) suspending agent, and (iv) stabilizer.
2. The method of claim 1 wherein the formulation is dry syrup containing free
flowing granules in a bottle, which is to be reconstituted at the time of use with boiled and cooled drinking water up to the mark given on the bottle.
3. The method of claim 1 wherein the digestive enzymes are fungal alpha amylase and
pepsin where label claim of formulation is Each 5 ml of reconstituted syrup contains Fungal alpha amylase derived from Aspergillus oryzae (1:1200) 50 mg and Pepsin (1:3000) 10 mg.
4. The method of claim 1 wherein the water soluble sweetener is Pharma grade
sucrose, incorporated in an amount of 50 to 95 weight parts per 100 weight parts of the granules.
5. The method of claim 1 wherein the suspending agent is natural gums, incorporated
in an amount of 0.5 to 1.5 weight parts per 100 weight parts of the granules.
6. The method of claim 1 wherein the stabilizer is Calcium salts, incorporated in an
amount of 0.01 to 1.0 weight parts per 100 weight parts of the granules.
7. The method of claim 1 wherein the reconstituted syrup has pH in the range of 3 -7.
8. The method of claim 1 wherein the moisture content is not more than 2 %.
9. The method of claim 1 wherein the granules are stable upto 2 years when stored at cool and dry place.
12

10. The method of claim 1 wherein the reconstituted syrup is stable for 15 days when stored at cool place.
Dated this 29th day of November 2005

Dr. Gopakumar G. Nair Agent for the Applicant
13

Abstract:
This invention provides a process for preparation of Dry Syrup containing digestive enzymes as free flowing granules having sufficient shelf life, comprising of (i) a pharmaceutically active digestive enzymes, (ii) water soluble sweetener, (iii) suspending agent, and (iv) stabilizer; which exhibits good stability and reconstitution into palatable syrup used for digestive disorders.
14

Documents:

1480-MUM-2005-ABSTRACT(29-11-2005).pdf

1480-MUM-2005-ABSTRACT(GRANTED)-(28-7-2011).pdf

1480-mum-2005-abstract.doc

1480-mum-2005-abstract.pdf

1480-MUM-2005-CANCELLED PAGES(27-6-2011).pdf

1480-MUM-2005-CLAIMS(16-9-2009).pdf

1480-MUM-2005-CLAIMS(29-11-2005).pdf

1480-MUM-2005-CLAIMS(AMENDED)-(16-9-2009).pdf

1480-MUM-2005-CLAIMS(AMENDED)-(26-5-2011).pdf

1480-MUM-2005-CLAIMS(AMENDED)-(27-6-2011).pdf

1480-MUM-2005-CLAIMS(GRANTED)-(28-7-2011).pdf

1480-MUM-2005-CLAIMS(MARKED COPY)-(27-6-2011).pdf

1480-mum-2005-claims.doc

1480-mum-2005-claims.pdf

1480-MUM-2005-CORRESPONDENCE(26-5-2011).pdf

1480-MUM-2005-CORRESPONDENCE(27-11-2007).pdf

1480-MUM-2005-CORRESPONDENCE(IPO)-(28-7-2011).pdf

1480-mum-2005-correspondence-received-ver-291105.pdf

1480-mum-2005-correspondence-received.pdf

1480-mum-2005-description (complete).pdf

1480-MUM-2005-DESCRIPTION(COMPLETE)-(16-9-2009).pdf

1480-MUM-2005-DESCRIPTION(COMPLETE)-(29-11-2005).pdf

1480-MUM-2005-DESCRIPTION(GRANTED)-(28-7-2011).pdf

1480-MUM-2005-FORM 1(16--9-2009).pdf

1480-MUM-2005-FORM 1(16-9-2009).pdf

1480-MUM-2005-FORM 1(27-6-2011).pdf

1480-MUM-2005-FORM 1(29-11-2005).pdf

1480-MUM-2005-FORM 18(27-11-2007).pdf

1480-mum-2005-form 2(16-9-2009).pdf

1480-MUM-2005-FORM 2(COMPLETE)-(29-11-2005).pdf

1480-MUM-2005-FORM 2(GRANTED)-(28-7-2011).pdf

1480-MUM-2005-FORM 2(TITLE PAGE)-(16-9-2009).pdf

1480-MUM-2005-FORM 2(TITLE PAGE)-(27-6-2011).pdf

1480-MUM-2005-FORM 2(TITLE PAGE)-(29-11-2005).pdf

1480-MUM-2005-FORM 2(TITLE PAGE)-(GRANTED)-(28-7-2011).pdf

1480-MUM-2005-FORM 3(16--9-2009).pdf

1480-MUM-2005-FORM 3(16-9-2009).pdf

1480-MUM-2005-FORM 3(26-5-2011).pdf

1480-MUM-2005-FORM 3(29-11-2005).pdf

1480-mum-2005-form-1.pdf

1480-mum-2005-form-2.doc

1480-mum-2005-form-2.pdf

1480-mum-2005-form-26.pdf

1480-mum-2005-form-3.pdf

1480-MUM-2005-REPLY TO FIRST EXAMINATION REPORT(16--9-2009).pdf

1480-MUM-2005-REPLY TO HEARING(27-6-2011).pdf


Patent Number 248622
Indian Patent Application Number 1480/MUM/2005
PG Journal Number 30/2011
Publication Date 29-Jul-2011
Grant Date 28-Jul-2011
Date of Filing 29-Nov-2005
Name of Patentee MERCURY LABORATORIES LIMITED
Applicant Address 2/13 & 2/14, GORWA INDUSTRIAL ESTATE, GORWA, BORADO -390 016
Inventors:
# Inventor's Name Inventor's Address
1 RAYASA RAMCHANDRA MURTHY SRINIVASA PHARMACY DEPARTMENT, FACULTY OF TECHNOLOGY AND ENGINEERING, THE M.S. UNIVERSTIY OF BORODA, BORODA- 390001
2 SHAH RANJENDRA RAMANLAL 2/13 & 2/14, GROWA INDUSTRIAL ESTATE, GORWA, BORODA - 390 016
3 PATIL JAIDEEP TRYAMBAK 2/13 & 2/14, GORWA INDUSTRAIL ESTATE, GORWA, BORADO -390 016
PCT International Classification Number A61K9/14
PCT International Application Number N/A
PCT International Filing date
PCT Conventions:
# PCT Application Number Date of Convention Priority Country
1 NA