enzymes, enzymes function

1. Enzyme Therapy
Submitted To:- Submiited By:-
Dr.Udita Tiwari Sachin Singh Rawat
HOD, Department of Biochemistry MSc. 2nd Sem

2. Learning objectives
Introduction
Properties of therapeutic enzymes
Limitions of enzymes
Enzymes used in Therapy
Enzymes Effectiveness
Conclusion
Reference

3. Introduction:-
• The role of enzymes in medicine as markers, diagnostic and prognostic applications
are widely known. These enzymes are now considered as new biological drug in the
treatment of certain disorders and hence emerged as an offshoot of therapy.
• More recently, enzymes have found the application as a new class of therapeutic
agents. This new emerging field named as enzymotherapy and will provide a new
class of biological durgs in the coming days.

4. • More than 200 enzymes deficiencies related to genetic dysfuction has been reported
in humans. Research in enzyme replacement therapy and gene therapy is currently in
progress throught the world, it will take considerable time for establishment of an
efficient gene therapy for each of these deficiencies. Till then enzyme therapy is an
intermediate solution.
• Role of enzymes as a replacement therapy in inherited genetic disease, as scavengers
to limit tissue damage during in inflammatory disease and in various ischemia /
reperfusion injuries, as specific detoxification agents during acute chemical
poisoning as a temporary analgesic agent during an acute organ failure and as
antineoplastic agents.

5. Properties of therapeutic enzymes:-
• The favored kinetic properties of the enzymes are low Km and high Vmax in order to
be maximally efficient even at low concentration of enzyme and substrate.
Therapeutic enzymes preparation are available for sale as lyophilized pure
preparations with biocompatible buffering salt and mannitol as diluents.

6. Limitations of enzymes:-
• Not all patient are suitable for treatment, some organs and tissues are corrected moe
readily than other, and there are problems with gauging efficacy in these highly
variable disorders.
• Finally, the therapies are expensive, limiting access to patients from those countries
that are able to afford expensive health care.

7. Enzymes used in Therapy:-
• Digestive enzymes
Pepsin
Cellulase
Chymotrypsin
Lactase
Lipase
• Deworming enzymes
• Debriding enzymes
• Streptokinase and Urokinase
• Adenosine Deaminase
• β-lactamase
• Asparaginase

8. Digestive enzymes
• Produced by the glands such as pencreas and secreted into the stomach and SI.
• Either due to inborn enzyme deficiency or due to acquired disease, the secretary glands
will not be able to secrete the enzymes at all or sometimes not to the level they are
required to digest the food.
• Sometimes in the case of permanent removal of pencreas due to pancreatitis.
• The patient requires digestive enzymes for rest of his life.
• α-Amylase:- Salivary α-amylase in mouth begins the hydrolysis of α-1,4 glucosidic bond
of the starch removing successive maltose units from the non-reducing end of the chain,
and the processis completed by the pencreatic amylase.

9. Pepsin:-
• Digestive enzyme found in gastric juice.
• That catalyzes the breakdown of protein to peptides in the middle of the polypeptide
chain.
• Pepsin supplement is used as a digestive aid to enhance body's natural metabolism
and remove the toxins from the intestine.
• Pepsin is usally prepared from the stomach of pigs.

10. Cellulase:-
• Cellulase is enzyme that breaks down cellulose to β-glucose, produced by bacterial
fungi and grazing animals such as cow.
• Cellulose is non digestible by human because we donot produce cellulase.
• Dietary supplement of cellulase helps to break down plant material better.

11. Chymotrypsin:-
• Chymotrypsin is a proteolytic enzymes naturally produced by the pancreas of the
human body.
• Chymotrypsin breaks down protein into dipeptide and some single amino acids by
hydrolysis of peptide bond in SI.
• It is selective for peptide bonds with aromatic or large hydrophobic side chains of the
carboxyl side chains.
• It has been taken as dietary supplement to improve health and digestion and aid in
treatment of various diseases.

12. Lactase
• Found in liver, kidney and intestine.
• Lactase breaks down lactose to glucose and galactose.
• A person suffering from the lactose deficiency may develop abnormal digestion of
milk protein.
• Lactose intoleranse may result in abdominal pain, cramps, gas, and wind in babies.

13. Lipase
• Pancreatic and intestinal lipase breakdown neutral fat into glycerol and fatty acids.
• A common symptom of lipase deficiency is the muscular spasms.
• Lipase usually derived

14. Deworming enzymes
• Plant proteases such as papain from papaya and ficin from fig are used for
deworming the gastrointestinal tract against the nematodes.
Debriding enzymes
• The enzymes effectively clean open wounds by removal of foreign matter and any
surrounding dead tissues.
• This allows rapid healing of the wound.
• The enzymes are applied only topically to the affected areas.
• Trypsin, papain, collagenase are the principal debriding enzymes.

15. Streptokinase and Urokinase
• Used to treat disorders within the circulatory system that lead to the formaton of
blood clots in the circulatory system.
• Streptokinase and urokinase are secreted by several species of streptococci which can
bind and activate human plasminogen to plasmin, by hydrolytic bond cleavage
breaks down to break down into fibrin.
• These enzymes can be used to dissolve medications in some cases of myocardial
infarction.

16. Adenosine deaminase
• ADA is cytoplasmic enzyme found in high concentration in lymphoid cells.
• Deficiency of ADA causes combined immunodeficiency disease due to
lymphocytotoxic effects of adenosine and deoxyadenosine.
• Deoxyadenosine cause dATP pool expansion, which blocks DNA replication by
inhibiting ribonucleotide reductase.
• Pathology of ADA deficiency is well defined and limited to haemopoietic cells, so
enzyme replacement therapy is straight-forward.
• Two problems associated with enzyme therapy are very short circulating life of the
injected enzyme and risk of provoking allergic or other immunologic response in
chronic treatment.
• Enzyme replacement therapy in ADA deficiency disease is aproved by US FDA.

17. β-lactamase
• The enzyme β-lactamase hydrolyzes the β-lactam ring of the penicillin to produce
pencilloate, which lacks antibacterial activity.
• Penicillin Penicilloate treating pencillin allergy.
• Hypersensitivity reaction is common in patients receiving penicillin and may appear
in the absence of prior exposure to drug.
• This reaction results in rash, fever and bronchospasm and in some individuals it may
lead to severe erythema multiforme accompanised by fever, headache, conjuctivitis
and arthralgia.
• The most frequent consequence of penicillin injection is the anaphylactic shock.

18. DNAs
• Cystic fibrosis (CF) is one of the most commonly occuring genetic diseases.
• Cystic fibrosis (CF), also known as mucoviscidosis, affects most critically the lungs,
and alsothe pancreas, liver and intestine
• It is characterized by abnormal transport of chloride and sodium across an
epithelium, leading to thick, viscous secretions i.e., Underlying cause is identified to
the mulfunction of ion transport
• Major clinical symptom is the production of viscousmucus in the respiratory track.
• Patients are susceptible to frequent lung infections and some patients develop
antibiotic resistance bacteria and hence, bacteria accumulate leading to a viscous
mucous secretion, clogging the bronchia and bronchioles.

19. • Thick mucous = alginate that is secreted by the bacteria + DNA released when
bacterial cells and degenerating leucocytes that accumulate in response to infection are
lysed The role of DNase I can hydrolyse long polymeric DNA chains into shorter
oligonucleotides and the purified enzyme can be delivered in an aerosol mist to the
lungs of CF patients to prevent respiratory distress.
• The enzyme could decrease the mucus viscosity in the lungs and allow patients for
easy breathing, thus reducing the severity and pain of the patient.
• This enzyme was approved for use by the US FDA in 1994

20. Lipase (Glucocerebrosidase)
• Gaucher's disease is one of the ten known hereditary lipid storage diseases doing
damage to the nervous system.
• Children suffering this disease lack enzyme which normally breakdown lipids in the
body, glucocerebroside.
• This material then accumulates in the liver, spleen, bones producing swellings,
causing damage to the nerves and death in children before reaching the age of two.
• Glucocerebrosidase is injected to cells enforged with lipid, the could be relieved of
the Gaucher's disease.
• At, present, the enzyme is purified from human urine or placenta where it is present
in small quantity.
• The major disadvantage of this enzymotherapy is its cost.

21. Asparaginase
• Asparaginase is a chemotherapy drug used to treat acute lymphoblastic leukaemia. It
can also be used to treat some other blood disorders.
• One form of asparaginase is made from E.coli & another form of asparaginase is
made from Erwinia chrysanthemi bacteria.
• Tumour cells are unable to synthesize L- asparagine due to the deficiency of
asparaginase and these components are crucial for the develoment and proliferation
of the cell.
• Asparaginase can be introduced intravenously that in turn drastically reduces the
level of free L- asparagine, creating a starvation in tumour cells for these amino
acids.
• Enzymes such as glutaminase, serine hydratase, arginase, and leucine dehydrate are
also tested for treating tumor cells

22. DMK Enzyme Therapy
• It is a professional treatment that addresses weak, prematured aged and inflamed
skin.
• It utilizes messenger enzymes to mimic the body's own chemistry, hydrolyse dead
cell material from the skin and remove impurities through a reverse osmosis process.
• This detoxifying back flushing of the skin increases circulation, bringing oxygenated
blood and nutrients to the skin, while improving lympahtic and enhancing collagen
production.

23. Alzheimer's disease
• Researcher are investigating how they might use an enzyme called BACE2 to treat
and possibly even prevent Alzheimer's disease.
• BACE2 has been destroy beta amyloid, a protein that accumulates in the brains of
individuals of with this form dementia.

24. Enzyme Effectiveness
• It is by the amount of activity level in the bloodstream after the enzyme has been
absorbed from the SI. Enzymes lose activity in low pH(1.0 -3.0) the stomach.
• Studies show that unprotected enzymes can lose up to 100% of their activity in 30
min or less.
• Enteric coating is the only reliable method to ensure that 100% of the enzymes are
not destroyed, and is by far the most effective delivery method to prevent release of
enzymes before they reach the SI.
Enzyme Effectiveness Determination

25. Uncoated capsules and tablets
• Stomach acid breaks down tablets and capsules and prematurely releases active
enzymes.
• The highly acidic environment of the stomach destroys the majority of enzymes
activity.
• If the tablet is of poor quality the may pass through both stomach and intestine with
no absorption.

26. Delivery system
• The real differentiator among Systemic Enzymes is in method of deliver to the
stomach and utimately to the SI.
• The only method to protect and effectively deliver a systemic enzyme is a 100 %
enteric-coated product.
• Protein molecules are normally destroyed by the hydrochloric acid of the stomach for
this reason tablets, capsules, or granules are used that are enteric-coated or covered
with an acid-proof layer.
• In this way the enzymes are held safely until the section of the intestines is reached
where their resorption is possible.

27. Oral Delivery Systems
1. Powder Filled Capsule
2. Tablet
3. Enteric Coated Liquid Capsule
4. Powder Filled Capsule

28. Powder Filled Capsule
Vegetarian or Gelatin Capsule:- Powder filled capsule technology is suitable for
contents that are not suspectible to the acidity of the stomach. The capsule and its
enzymes are desolved prematurely in the stomach, resulting in up to 100% destruction
of the enzymes within 30 minutes in the extremely low pH environment.

29. Tablet Digestion
Enteric Coated Tablet:- Enteric coated tabletsare able to withstand stomach acid. They
must be introduced into a high pH environment for extended periods before dissolving.
Since individual pH levels vary, the tablet may dissolve very late in the SI or bypass it
completely. Many users also complain of synthetic coating.

30. Liquid Filled Gel Capsule
Gel Capsule Digestion:- Enteric Coated Liquid-Filled Gel Capsule: The all-natural
protective coating prevents the enzymes from being destroyed by the stomach acids.
The pancreatic juices break down the coating in the duodenum. This makes 100% of the
enzyme contents available for absorption into the bloodstream.
Enteric Coating: Enteric Coating is the best and most effective oral delivery system
today for systemic enzymes. The technology of enteric coated liquid-filled capsules is
superior to that of powdered enzymes because it uses low heat production, provides
more uniform dose distribution, and prolongs enzyme activity.

31. Therapeutic enzymes
Enzymes Therapeutic application
Asperginase Leukemia
Lysozymes Antibiotic
Uricase Gout
Urease Renal failure
Tyrosinase Liver failure

32. Conclusion
Enzyme therapy has improved the lives of many patients, by reducing the burden of
their disease. Several studies have sought to determine to what extent optimal clinical
outcomes are a function of the prescribed enzyme dose and its resultant costs. Goals of
treatment should include important clinical end points, such as enhanced quality of life
and reduced the diseases.

33. Reference
• S.N. JOGDAND, Medical biotechnology, Himalaya publishing house page no 43-48
and 138-145
• Prathibha Nallari, V.Venugopal rao, Medical biotechnology, oxford university press
2010, page no 89-95