Download Therapeutic Enzymes

In The Name Of God
The Most Compassionate
The Most Merciful
Therapeutic Enzymes
By
Ali Jahanian Najafabadi
Doctor of pharmacy (Pharm. D.)
Ph.D of Pharmaceutical Biotechnology
Introduction
Enzymes: definition
Enzymes are the catalysts of biological processes
Enzymes speed up the rate of biochemical reactions by several
orders of magnitude and function under relatively mild conditions of
pH and temperature
Several thousand enzymes have been identified to date
Today, a myriad enzymes are employed for industrial, analytical,
and therapeutic purposes
Introduction
Enzymes: General features
Almost all enzymes are protein-based
They are globular proteins
soluble in aqueous-based solutions
Sensitive to environmental conditions such as changes in temperature
and pH
Enzymes are high-molecular mass molecules, ranging from 13 to 500
kDa, although many fall within the 30 to 100 kDa range
They generally display a high degree of specificity with respect to the
substrate(s) with which they interact
The region of the enzyme where the substrate is transformed to form the
product is known as the active site
Some enzymes also require the presence of a nonprotein cofactor at the
active site in order to maintain catalytic activity
Introduction
Applied Enzymology
Today, enzymes find a broad range of applications within:
•Industry
•Medicine
•Analytical reagents
Introduction
Applied Enzymology
Therapeutic Enzymes
Applications of therapeutic enzymes
To replace enzymes which are missing or defective as a
consequence of inherited genetic disease
To replace enzymes that are deficient or present only in inadequate
quantity as a consequence of acquired disease in the organ(s) where
they are normally synthesized
To provide a specific biological effect which is dependent on the
catalytic activity of the enzyme
Therapeutic Enzymes
Requirements of effective therapeutic enzymes
They must reach their site of action in the body and tissue
compartment
They must be active under the conditions present at the intended site
of action. This includes substrate and cosubstrate/coenzyme availability,
appropriate redox potential, adequate pH value for activity, and absence
(or saturation) of inhibitors.
They must be sufficiently stable to ensure adequate pharmacokinetics,
i.e., the required activity level for the necessary time period
They must be sufficiently soluble to allow application as a solution if
administration is via the intravenous, intramuscular, or subcutaneous
route
Therapeutic Enzymes
Requirements of effective therapeutic enzymes (continued)
They must be sufficiently pure to avoid eliciting unwanted side
reactions caused by contaminants, e.g., microbial endotoxins,
pyrogens, or other harmful materials
Therapeutic Enzymes
Challenges remain for the future development of therapeutic enzymes
Improving stability
Coupling of enzymes to polyethylene glycol (PEGylation)
Chemical cross-linking
Development of sustained-release forms and depot systems
e.g., by microencapsulation of the enzyme in polymers such as
poly(lactide)– poly(glycolide) copolymers, or by entrapment
within artificial liposomes or red blood cell ghosts; as well as the
removal of receptor binding sites by designing appropriate
enzyme muteins.
Therapeutic Enzymes
Challenges remain for the future development of therapeutic enzymes
Improve tissue penetration
Decreasing molecule size (muteins)
Fusion to sequences mediating membrane permeation
TAT protein from human immunodeficiency virus-1
VP22 protein from Herpes simplex virus
Therapeutic Enzymes
Challenges remain for the future development of therapeutic enzymes
Diminish immunogenicity
By chemical (PEGylation) or genetic modification to cover or
remove epitopes on the protein surface recognized by the
immune system
Achieve specific targeting to tissues/cells
Modification with sugar residues which are recognized by
hepatocyte receptors
Genetic fusion to ‘‘targeting devices’’ such as recognition
peptides or antibody fragments
Therapeutic Enzymes
Sources and production of therapeutic enzymes
For many decades, enzymes obtained from natural nonhuman
sources such as bacteria, fungi, animals, and snake venoms have
been used for therapeutic purposes
Due to their immunogenicity, these enzymes usually are suitable
only for external applications (e.g., digestive or topical)
Therapeutic Enzymes
Sources and production of therapeutic enzymes (continued)
A number of human enzymes obtained from blood, urine, placenta,
or human cell culture have been introduced into clinical practice, such
as blood fractions and coagulation factors, urokinase, alglucerase,
and others
Limited supply of starting material
Reliable elimination of viral contamination
Since the 1980s, human enzymes are preferentially obtained by
recombinant DNA technology
Therapeutic Enzymes
Sources and production of therapeutic enzymes (continued)
Whereas
initially
basic
replacement
protein
products,
i.e.,
recombinant proteins identical with the native human enzymes
were developed, there is now a trend towards engineered ‘‘secondgeneration’’ protein products whose amino acid sequence has been
deliberately altered to improve properties relevant for therapeutic
efficacy or safety, or to improve the manufacturing process
Therapeutic Enzymes
Sources and production of therapeutic enzymes (continued)
Expression systems used for production of recombinant enzymes
Human cell lines
Mammalian cell lines (CHO, BHK)
Yeast
E. coli
Expression by transgenic plants or in the milk of transgenic
animals
Therapeutic Enzymes
Engineering therapeutic enzymes
In addition to engineering the traditional features of enzymes, such
as stability, specificity, and catalytic activity, engineering
therapeutic enzymes aims to improve:
•Serum half life
•Efficacy of different therapeutic mechanisms
•Potential for deleterious side effects and increased immunogenicity
•Endogenous protease susceptibility,
•Create a faster or slower acting product
Therapeutic Enzymes
Engineering therapeutic enzymes
Therapeutic enzymes engineering can involve:
•Alteration of the enzyme’s primary aa sequence
•Modification of the glycocomponent of a glycosylated protein
•Covalent attachment of chemical entities such as PEG
Therapeutic enzymes
at a glance
Therapeutic Enzymes
Enzymes used therapeutically
Therapeutic Enzymes
Asparaginase
In the late 1970s, researchers illustrated that serum transferred from healthy
guinea pigs into mice suffering from leukemia contained some agent capable of
inhibiting the proliferation of the leukaemic cells
A search revealed the agent to be asparaginase
Asparaginase is an enzyme capable of catalysing the hydrolysis of Lasparagine, yielding aspartic acid and ammonia
Therapeutic Enzymes
Asparaginase
Most healthy (untransformed) mammalian cells are capable of directly
synthesizing asparagine from glutamine
Hence, asparagine is generally classified as a non-essential amino acid
However, many transformed cells lose the ability to synthesize asparagine
themselves. For these, asparagine becomes an essential amino acid
Therapeutic Enzymes
Asparaginase
In the case of the leukaemic mice, the guinea pigs’ asparaginase deprived the
transformed cells of this amino acid by hydrolysing plasma asparagines
This approach has been successfully applied to treating some forms of human
leukemia
Sources: E. coli , Erwinia chrysanthemi, Pseudomonas and Acinetobacter
Therapeutic Enzymes
Asparaginase
Trade name: Oncaspar™ (pegaspargase)
Approval Date: 1994
Description: Pegaspargase is a modified version of the antineoplastic enzyme
L-asparaginase. L-asparaginase is modified by covalently conjugating units of
monomethoxypolyethylene glycol (PEG), molecular weight of 5kDa, to the
enzyme, forming the active ingredient PEG-L-asparaginase.
Production system: Escherichia coli
Dosage form: a solution (1 mg active/ml) to be administered by inhalation with a
nebulizer system
Therapeutic Indications: Treatment of patients with acute lymphoblastic
leukemia (ALL) who are hypersensitive to native forms of Lasparaginase
Therapeutic Enzymes
Asparaginase
Trade name: Erwinaze™ (asparaginase from Erwinia chrysanthemi)
Approval Date: 2011
Description: contains an asparaginase specific enzyme derived from Erwinia
chrysanthemi. L-asparaginase is a tetrameric enzyme consisting of four identical
subunits, each having a molecular weight of about 35 kDa.
Production system: Erwinia chrysanthemi
Dosage form: Each vial contains 10,000 International Units of asparaginase
Erwinia chrysanthemi
Therapeutic Indications: is indicated as a component of a multi-agent
chemotherapeutic regimen for the treatment of patients with acute lymphoblastic
leukemia (ALL) who have developed hypersensitivity to E. coli-derived
asparaginase
Therapeutic Enzymes
Asparaginase
side effects: severe nausea, vomiting and diarrhoea and compromised liver
and kidney function
Side effects are probably due to a transient asparaginase deficiency in various
tissues
Under normal circumstances, dietary-derived plasma asparagine levels are
sufficient to meet normal tissue demands, and the cellular asparagine biosynthetic
pathway remains repressed.
Therapeutic Enzymes
Asparaginase
Reduced plasma asparagine levels result in the induction of cellular asparagine
synthesis. High-dose asparaginase administration will immediately reduce plasma
asparagine levels. However, the ensuing initiation of cellular asparagine synthesis
may not occur for several hours. Thus, a more suitable therapeutic regimen may
entail initial low-dose asparaginase administration, followed by stepwise
increasing dosage levels
Therapeutic Enzymes
Dornase alfa
Recombinant DNase preparations have been used in the treatment of cystic
fibrosis since the end of 1993
A number of clinical symptoms characterize cystic fibrosis. Predominant
among these is the presence of excess sodium chloride in cystic fibrosis
patient sweat. Indeed, measurement of chloride levels in sweat remains the
major diagnostic indicator of this disease
The production of an extremely viscous, custard-like mucus in various body
glands/organs that severely compromises their function
Therapeutic Enzymes
Dornase alfa
Particularly affected organs are:
The pancreas: the mucus blocks its ducts in 85 per cent of cystic fibrosis
patients
The reproductive tract: which can render males subfertile or infertile
The liver: in which bile ducts can become clogged
The small intestine: can become obstructed by mucus mixed with digesta
Therapeutic Enzymes
Dornase alfa
Particularly affected organs are:
The lungs: which mucus compromises respiratory function. Also render this
tissue susceptible to frequent and recurrent microbial infection which leads
to immune reactions. Finally large quantities of DNA are released from
damaged microbes and neutrophils at the site of infection. High molecular
mass DNA is itself extremely viscous and increases substantially the viscosity
of the respiratory mucus
Therapeutic Enzymes
Dornase alfa
The genetic basis of this disease:
Mutations in a gene located on chromosome7
This gene is expressed largely by cells present in sweat glands, the lung,
pancreas, intestine and reproductive tract
70 percent of all cystic fibrosis patients exhibit a specific three-base-pair
deletion in the gene, which results in the loss of a single amino acid
(phenylalanine 508) from its final polypeptide product
The gene product is termed cystic fibrosis transmembrane conductance
regulator (CFTR), and it codes for a chloride ion channel
The relatively recent innovation in cystic fibrosis therapy is the use of DNase
to reduce the viscosity of respiratory mucus
Therapeutic Enzymes
Dornase alfa
Trade name: Pulmozyme™
Approval Date: 1993 (U.S.)
Description: it is a 37-kDa, 260-amino acid recombinant human
deoxyribonuclease I (DNase I) that selectively cleaves DNA
Production system: Chinese hamster ovary (CHO)
Dosage form: a solution (1 mg active/ml) to be administered by inhalation with a
nebulizer system
Therapeutic Indications: cystic fibrosis
Therapeutic Enzymes
Dornase alfa
 The most common side effects:
Voice alteration
Rhinitis
Pharyngitis
Laryngitis
Rash
Therapeutic Enzymes
Imiglucerase
Gaucher's disease is an autosomal-recessive genetic disorder and is the most
common of the lysosomal storage diseases resulting from a deficiency of the
lysosomal enzyme glucocerebrosidase which affects lipid metabolism
Glucocerebrosidase: a lysosomal hydrolase
Glucosylceramide  glucose and ceramide
The disease is characterized by accumulation of glucocerebroside in tissue
macrophages, which become enlarged and are generally termed Gaucher's
cells
Gaucher's cells accumulate in spleen, liver, and bone marrow, resulting in
anemia, thrombocytopenia and sometimes mental retardation
Therapeutic Enzymes
Imiglucerase
Therapeutic Enzymes
Imiglucerase
Trade name: Cerezyme™
Approval Date: 1994 (U.S.), 1997 (E.U.)
Description: is a recombinant modified (macrophage targeted) human bglucocerebrosidase enzyme
Production system: Chinese hamster ovary (CHO)
Dosage form: lyophilized form, (200 IU activity per vial) to be reconstituted and
diluted before administration by intravenous infusion
Therapeutic Indications: Gaucher's disease
Therapeutic Enzymes
Imiglucerase
Cerezyme demonstrates an altered glycosylation pattern: The manufacturing
process includes an in vitro, post-translational enzymatic processing step using an
exoglycosidase
The native enzyme’s sugar side-chains are capped with a terminal sialic acid
or galactose residue which results in removing of more than 95 percent of
injected glucocerebrosidase from the circulation by the liver via binding to
hepatocyte surface lectins. Thus, the intact enzyme is not available for uptake by
the tissue macrophages (which display high levels of surface mannose receptors)
Therapeutic Enzymes
Imiglucerase
Treatment of native glucocerebrosidase with exoglycosidases, removes terminal
sugar residues, and exposes mannose residues of their sugar side-chains,
resulting in their binding to and uptake by the macrophages. In this way, the
‘mannose-engineered’ enzyme is selectively targeted to the affected cells.
Therapeutic Enzymes
Imiglucerase
Side effects:
Rare severe hypersensitivity reactions
pulmonary hypertension
anaphylactoid reactions
Tachycardia
Cyanosis
Cerezyme is contraindicated during pregnancy and lactation
Therapeutic Enzymes
TALIGLUCERASE ALFA
Trade name: ELELYSO™
Approval Date: 2012
Description: It is a monomeric glycoprotein containing 4 N-linked glycosylation
sites (Mr = 60,800). Taliglucerase alfa differs from native human βglucocerebrosidase by two amino acids at the N terminal and up to 7 amino
acids at the C terminal. Taliglucerase alfa is a glycosylated protein with
oligosaccharide chains at the glycosylation sites having terminal mannose sugars
Production system: carrot plant root cells
Dosage form: A sterile, non-pyrogenic, lyophilized product
Therapeutic Indications: Long-term enzyme replacement therapy (ERT) for
adults with a confirmed diagnosis of Type 1 Gaucher disease
Therapeutic Enzymes
Laronidase
Mucopolysaccharidoses
are
rare,
inherited
genetic
diseases
characterized
by
incomplete degradation of specific mucopolysaccharides in the body
Mucopolysaccharides (glycosaminoglycans) refer to the polysaccharide side chains of
a class of biological molecules known as proteoglycans
When dissolved in water, proteoglycans form very viscous solutions
They usually function to support fibrous and cellular elements of tissue and help
maintain water and salt balance in the body
Therapeutic Enzymes
Laronidase
The major glycosaminoglycans found in the body include
Hyaluronic acid
Chondroitin sulphate
Keratan sulphate
Dermatan sulphate
Heparan sulphate
Heparin
Mucopolysaccharidoses are characterized by the accumulation of the oligosaccharide
component of proteoglycans due to a deficiency in one or more of the lysosomal
hydrolysases normally responsible for degrading these molecules
Therapeutic Enzymes
Laronidase
MPS I patients
Therapeutic Enzymes
Laronidase
Mucopolysaccharidosis I (MPS I) is characterized by a deficiency of α-Liduronidase, which functions to catalyse the hydrolysis of the terminal α-Liduronic acid residue from dermatan sulphate and heparan sulphate
As a consequence, both dermatan
and heparan sulphate accumulate
throughout the body, leading to widespread organ and tissue dysfunction
Therapeutic Enzymes
Laronidase
Trade name: Aldurazyme™
Approval Date: 2003 (U.S.)
Description: Laronidase is a polymorphic variant of human α–L-iduronidase, a
lysosomal hydrolase that catalyses the hydrolysis of terminal α -L-iduronic acid
residues of dermatan sulphate and heparan sulphate. The mature enzyme is a
628-amino acid, 83- kDa monomeric glycoprotein, containing 6 N-linked
oligosaccharide side chains
Production system: Chinese hamster ovary (CHO)
Dosage form: a concentrate solution for intravenous infusion. The laronidase
concentration is 0.58 mg/ml, with a specific activity of approximately 172 U/mg
Therapeutic Indications: Mucopolysaccharidosis I
Therapeutic Enzymes
Laronidase
Two of the recombinant enzyme’s six oligosaccharide side chains terminate in
mannose-6-phosphate residues therefore:
Much of the enzyme administered is taken up by cells and targeted to
lysosomes by binding to specific mannose-6-phosphate receptors
Therapeutic Enzymes
Laronidase
Side effects:
Infusion-related hypersensitivity reactions
Upper-respiratory tract infections
Rash
Injection-site reactions
It has not been determined if the drug is excreted in human milk
Therapeutic Enzymes
Agalsidase beta
Fabry's disease is a genetic disease characterized by the deficiency of the
lysosomal enzyme α-galactosidase A, leading to an inability to break down
certain glycolipids, particularly the globotriaosylceramide (GB3)
The result of the disorder is an accumulation of glycolipids in the walls of
vascular cells, particularly in the kidney, heart, and nervous system
Therapeutic Enzymes
Agalsidase beta
Therapeutic Enzymes
Agalsidase beta
Fabrazyme proved its efficacy in the treatment of Fabry's disease with significant
reduction of GB-3 in the vascular endothelium of kidney, skin, and heart tissue
Due to the rarity of the disease (an estimated 500 to 1000 patients in the E.U.)
and the nature of the medicinal product, approval was granted after only
preliminary studies, and long-term clinical trials are still ongoing. No other
curative treatment is available for patients with Fabry's disease.
Most of the patients developed antibodies against Fabrazyme, but no reduction
in efficacy was observed
In order to prevent organ damage that could be difficult to reverse,
administration of Fabrazyme before the disease develops should be
considered
Therapeutic Enzymes
Agalsidase beta
Trade name: Fabrazyme™
Approval Date: 2001 (E.U.)
Description: it is a recombinant human α-galactosidase A. The 429 amino acid
glycoprotein spontaneously dimerizes, yielding the 100-kDa biologically active
enzyme
Production system: Chinese hamster ovary (CHO)
Dosage form: a lyophilized form (35 mg/vial) to be reconstituted and diluted
before use as an intravenous infusion
Therapeutic Indications: long-term ERT of Fabry's disease
Therapeutic Enzymes
Agalsidase beta
Side effects:
Chills and fever
Infusion-related reactions
Hypersensitivity
Cardiovascular symptoms
Gastrointestinal symptoms
Fabrazyme should not be administered with chloroquine, amiodarone, benoquin,
or gentamicin.
No safety and efficacy studies were carried out on patients under 16 or over 65
years old.
Fabrazyme is contraindicated during pregnancy and lactation
Therapeutic Enzymes
Agalsidase alpha
Trade name: Replagal™
Approval Date: 2001 (E.U.)
Description: it is a recombinant human a-galactosidase A, a lysosomal
hydrolase. The enzyme is a glycosylated homodimer, with each 50-kDa, 398amino acid subunit containing three N-linked oligosaccharides
Production system: continues human cell line
Dosage form: a concentrated solution (1 mg/ml) for intravenous infusion over
40 minutes
Therapeutic Indications: long-term ERT of Fabry's disease
Therapeutic Enzymes
Rasburicase
Uric acid is the end-product of purine metabolism in humans, other primates,
birds and reptiles
It is produced in the liver by the oxidation of xanthine and hypoxanthine
Humans do not break down uric acid, which is instead removed by the kidney
In the presence of high levels of uric acid, crystals precipitate in the kidney,
leading gout or urate stones in the urinary tract and finally to severe renal failure
Therapeutic Enzymes
Rasburicase
Significantly elevated serum uric acid concentrations can also be associated with
rapidly proliferating cancers or, in particular, with onset of chemotherapy
Purine metabolism in some mammals is characterized by a further oxidation of
uric acid to allantoin by the enzyme urate oxidase
Allantoin is significantly more water soluble than uric acid and is also freely
excreted via the renal route
Therapeutic Enzymes
Rasburicase
Summary overview of purine metabolism
Therapeutic Enzymes
Rasburicase
Trade name: Fasturtec™ (trade name E.U.), Elitek™ (trade name U.S.)
Approval Date: 2001 (E.U.); 2002 (U.S.)
Description: it is a recombinant form of the Aspergillus flavus urate oxidase,
the enzyme that converts uric acid to allantoin. The biologically active form of the
enzyme is a tetramer, consisting of four identical (34-kDa, 301- amino acid)
polypeptides
Production system: S. cerevisiae
Dosage form: a lyophilized form (1.5 mg/vial) to be reconstituted and diluted
before use as an intravenous infusion
Therapeutic Indications: treatment and prophylaxis of acute hyperuricaemia to
prevent acute renal failure in patients with hematological malignancy with a high tumor
burden and patients at high risk of a rapid tumor lysis or shrinkage at initiation of
chemotherapy
Therapeutic Enzymes
Rasburicase
Side effects
Fever
Nausea
Vomiting
Allergic-type reactions and the production of antirasburicase antibodies
 Fasturtec should not be readministered
It should not be administered in case of metabolic disorders such as G6PD
deficiency, because the production of H2O2 as a product of degradation of uric
acid, if not eliminated, could cause hemolytic anemia
Fasturtec is contraindicated during pregnancy and lactation
Therapeutic Enzymes
Superoxide dismutase
Under normal circumstances in aerobic metabolism, oxygen is reduced by four
electrons, forming H2O. Although this usually occurs uneventfully, incomplete
reduction will result in the generation of oxygen radicals and other reactive
species. These are: the superoxide radical O2- , hydrogen peroxide (H2O2) and
the hydroxyl radical (OH-)
The superoxide and hydroxyl radicals are particularly reactive and can attack membrane
components, nucleic acids and other cellular macromolecules, leading to their
destruction/modification
O2- and OH- radicals are believed to be amongst the most mutagenic substances
generated by ionizing radiation
Therapeutic Enzymes
Superoxide dismutase
Oxygen-utilizing organisms have generally evolved specific enzyme-mediated
systems that serve to protect the cell from such reactive species. These enzymes
include SOD and catalase or glutathione peroxidase (GSH-px), which catalyse
the following reactions:
Therapeutic Enzymes
Superoxide dismutase
three types of SOD have been identified:
A cytosolic eukaryotic dismutase, generally a 31 kD dimer, containing both
copper and zinc
At least three types of SOD have been identifi ed:; a 75 kD mithocondrial
form and a 40 kD bacterial form, each of which contains two manganese
atoms
An iron-containing form found in some bacteria, blue–green algae and many
plants
The metal ions play a direct role in the catalytic conversion, serving as transient
acceptors/donors of electrons
Therapeutic Enzymes
Superoxide dismutase
In humans, increased generation of O2 and/or reduced SOD levels have been
implicated in a wide range of pathological conditions including:
Ageing
Asthma
Accelerated tumour growth
Neurodegenerative diseases
Inflammatorytissue necrosis
SOD isolated from bovine liver or erythrocytes has been used medically as an
anti-inflammatory agent. Human SOD has also been expressed in several
recombinant systems, and is currently being evaluated to assess its ability to
prevent tissue damage induced by exposure to excessively oxygen-rich blood.
Therapeutic Enzymes
Debriding agents
Debridement refers to the process of cleaning a wound by removal of foreign
material and dead tissue
Cleansing of the wound facilitates rapid healing and minimizes the risk of
infection due to the presence of bacteria at the wound surface
The formation of a clot, followed by a scab, on a wound surface can trap
bacteria, which then multiply (usually evidenced by the production of pus), slowing
the healing process
Although debridement may be undertaken by physical means (e.g. cutting away
dead tissue, washing/cleaning the wound), proteolytic enzymes are also often
used to facilitate this process
Trypsin, papain, collagenase and various microbial enzymes have been used in
this regard.
Therapeutic Enzymes
Debriding agents
Trypsin: a 24 kDa proteolytic enzyme synthesized by the mammalian pancreas
in an inactive zymogen form: trypsinogen
Upon its release into the small intestine, it is proteolytically converted into trypsin
by an enteropeptidase
Active trypsin plays a digestive role, hydrolysing peptide bonds in which the
carboxyl group has been contributed by an arginine or lysine
Trypsin used medically is generally obtained by the enzymatic activation of
trypsinogen, extracted from the pancreatic tissue of slaughterhouse animals.
Therapeutic Enzymes
Debriding agents
Papain: a cysteine protease isolated from the latex of the immature fruit and
leaves of the plant Carica papaya
It consists of a single 23.4 kda, 212 amino acid polypeptide, and the purifi ed
enzyme exhibits broad proteolytic activity
Collagenase: a protease that can utilize collagen as a substrate. Although it can
be produced by animal cell culture, certain microorganisms also produce this
enzyme, most notably certain species of Clostridia (the ability of these pathogens
to produce collagenase facilitates their rapid spread throughout the body)
Collagenase used therapeutically is usually obtained from cell fermentation
supernatants of Clostridium histolyticum. Such preparations are applied topically
to promote debridement of wounds, skin ulcers and burns
Therapeutic Enzymes
Debriding agents
Papain: a cysteine protease isolated from the latex of the immature fruit and
leaves of the plant Carica papaya
It consists of a single 23.4 kda, 212 amino acid polypeptide, and the purifi ed
enzyme exhibits broad proteolytic activity
Collagenase: a protease that can utilize collagen as a substrate. Although it can
be produced by animal cell culture, certain microorganisms also produce this
enzyme, most notably certain species of Clostridia (the ability of these pathogens
to produce collagenase facilitates their rapid spread throughout the body)
Collagenase used therapeutically is usually obtained from cell fermentation
supernatants of Clostridium histolyticum. Such preparations are applied topically
to promote debridement of wounds, skin ulcers and burns
Therapeutic Enzymes
Digestive enzyme
A number of enzymes may be used as digestive aids. In some instances, a
single enzymatic activity is utilized, whereas other preparations contain multiple
enzyme activities
These enzyme preparations may be used to supplement normal digestive
activity, or to confer upon an individual a new digestive capability
Therapeutic Enzymes
Digestive enzyme
The majority have been derived from animal (pancreatic extract), microbial
(e.g., Aspergillus oryzae), and plant (e.g., barley) sources
As the product never enters the blood stream, the product purity need not be
as stringent as enzymes (or other proteins) administered intravenously.
Most digestive enzymes are, at best, semi-pure preparations
Therapeutic Enzymes
Digestive enzyme
Amylase are enzymes that catalyze the hydrolysis of α(1-4)-glycosidic linkages
of polysaccharides such as starch and glycogen
Amylases are secreted by the pancreas and salivary glands in humans
Amylase is administered as a digestive aid to improve digestion of dietary
carbohydrate
Therapeutic Enzymes
Digestive enzyme
Cellulase is not produced by humans and is administered as a digestive
supplement to alleviate flatulence and to improve overall digestion, especially in
high-fiber diets
Cellulase can be derived from fungal (Aspergillus niger) and bacterial
(Bacillus) sources
Therapeutic Enzymes
Digestive enzyme
Invertase is utilized as a digestive aid in alleviating the symptoms of sucrase–
isomaltase deficiency.
Invertase alleviates symptoms by hydrolyzing sucrose to glucose and
fructose, which are absorbed to portal blood
The enzyme can be derived from Aspergillus or Saccharomyces species
Therapeutic Enzymes
Digestive enzyme
α-Galactosidase, which is not produce by the body, catalyzes the hydrolysis of
the α(1-6) linkages in α-galactoside carbohydrates such as melibiose and
raffinose.
The enzyme can be derived from selected strains of Aspergillus niger and
Saccharomyces cerevisiae.
α-Galactoside carbohydrates are widely found in legumes and cruciferous
vegetables
including
beans,
peas,
broccoli,
and
cabbage.
These
carbohydrates are fermented by bacteria in the colon, with the accompanying
production of gas.
Supplemental α-galactosidase catabolizes the oligiosaccharides prior to
reaching the colon and prevents flatulence and the symptoms associated with it
Therapeutic Enzymes
Digestive enzyme
Papain is a highly active plant protease which has a broad range of proteolytic
activity and is less expensive to produce than microbial enzymes
Pepsin is derived from the zymogen pepsinogen, which is secreted by gastric
mucosal cells, and it hydrolyzes dietary protein to shorter polypeptides and
peptides. Pepsin used as a digestive aid is typically derived from the gastric
mucosa of slaughterhouse animals
Bromelain is a complex natural mixture of proteolytic enzymes derived from
pineapple stems. Bromelain is used as a digestive aid to promote healthy
digestion by assisting in the hydrolysis of dietary protein
Therapeutic Enzymes
Digestive enzyme
β-galactosidase or lactase, hydrolyzes lactose, forming glucose and
galactose.
Lactose, or milk sugar, is a disaccharide found in the milk of most mammals
Lactose intolerance and infantile colic are two lactose-related conditions
whose symptoms can be relieved by dietary hydrolysis of the lactose load via the
use of exogenous β-galactosidase enzyme
Therapeutic Enzymes
Digestive enzyme
Pancreatin is a pancreatic extract usually obtained from the pancrease of
slaughterhouse animals
Pancreatic enzymes are essential for digestion of macromolecules such as
proteins, fats, and carbohydrates in the small intestine
Pancreatic enzyme supplementation may be deemed necessary in conditions
such as chronic pancreatitis, pancreatic carcinoma, gastric surgery, and
cystic fibrosis
Pancreatin is a pancreatic enzyme preparation derived from porcine pancreas.
Pancreatin comprises the pancreatic enzymes trypsin, amylase, nucleases, and
lipase
Therapeutic Enzymes
Digestive enzyme
Enzymes that are used as digestive aids
Bioreactors
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