Debriding composition from bromelain and methods of production thereof
Inventors
Assignees
Publication Number
US-8119124-B2
Publication Date
2012-02-21
Expiration Date
2025-11-22
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Abstract
The present invention relates to a debriding composition obtained from bromelain and to methods of producing same. Particularly, the present invention relates to a debriding composition obtained from bromelain comprising proteolytic enzymes having molecular weights of about 23 kDa, being essentially devoid of bromelain inhibitors, and to pharmaceutical compositions comprising same. The debriding compositions and the pharmaceutical compositions comprising same are particularly useful in debriding eschar tissues and in wound healing.
Core Innovation
The invention provides a debriding composition obtained from bromelain that comprises proteolytic enzymes with molecular weights of about 23 kDa and is essentially devoid of bromelain inhibitors. This composition is highly efficient in debriding non-viable tissues such as eschar tissues and promotes wound healing due to its distinct biochemical features and reproducible protein content.
The problem addressed by the invention stems from prior debriding preparations being unsatisfactory in selectively removing devitalized tissue without harming viable tissue. Previous proteolytic enzyme preparations, including purified enzymes and mixtures obtained from bromelain, suffered from issues of poor efficacy, toxic side effects, lack of selectivity, and presence of bromelain inhibitors, which reduced their debriding activity. There existed an unmet need for a biochemically characterized, reproducible, and inhibitor-free enzyme mixture from bromelain capable of efficient wound debridement.
The invention also discloses novel methods to produce such a debriding composition by combining steps including acidic extraction of bromelain with an antioxidant and filter aid addition, filtration to remove insoluble components, ammonium sulfate precipitation, pH adjustment, centrifugation, dissolution in acidic solution with antioxidant, and filtration using a membrane to retain proteolytic enzymes above about 10 kDa. The product consists predominantly of proteins of about 23 kDa molecular weight, is substantially free of bromelain inhibitors, and can be formulated into pharmaceutical compositions suitable for topical application to wounds for treatment.
Claims Coverage
The patent contains several independent claims covering compositions, methods for obtaining the composition, and methods of treatment. The main inventive features relate to the specific debriding composition from bromelain, its biochemical properties, methods of production including purification steps, and therapeutic uses.
Debriding composition with defined proteolytic enzymes and low inhibitor content
A debriding composition obtained from bromelain comprising a plurality of different proteolytic enzymes having molecular weights of about 23 kDa, including stem bromelain (SEQ ID No. 1 and 3), ananain (SEQ ID No. 2), and cysteine proteinase precursor (SEQ ID No. 4), wherein bromelain inhibitors are present in less than 10% w/w of protein content.
Method for obtaining the debriding composition with purification and filtration steps
A method comprising suspending bromelain in an acidic solution optionally with antioxidant (pH 2.4-4), adjusting pH, adding a silica filter aid, filtering to remove insolubles, precipitating proteolytic enzymes with ammonium sulfate at 40-50% saturation, incubating at 3-10°C, centrifuging to yield precipitate, dissolving precipitate in acidic solution with optional antioxidant, filtering to retain proteolytic enzymes over 10 kDa, and lyophilizing to obtain the debriding composition.
Method of treating wounds by applying the debriding composition
A method of treating wounds by debriding non-viable tissues comprising applying a therapeutically effective amount of the debriding composition as defined above, optionally including a pharmaceutically acceptable carrier, to wounds including burn wounds, ulcers, surgical wounds, traumatic wounds, infections, and scar tissue.
The claims cover the specific bromelain-derived debriding composition characterized by about 23 kDa proteolytic enzymes with minimal bromelain inhibitors, methods for producing this composition involving specific purification steps, and therapeutic methods for wound debridement and healing using this composition.
Stated Advantages
The debriding composition is highly efficient in debriding non-viable tissues compared to crude bromelain or other proteolytic mixtures.
The composition is essentially devoid of bromelain inhibitors, which contributes to improved activity and reproducibility.
The purified enzyme mixture has distinct and reproducible biochemical features advantageous for clinical use.
The composition avoids the toxic side effects and poor efficacy associated with previously used purified proteolytic enzymes.
Documented Applications
Debridement of eschar tissues associated with wounds.
Wound healing applications including partial and full thickness burns, sunburns, frostbite, ulcerative lesions such as pressure ulcers, varicose ulcers, stasis and trophic ulcers.
Treatment of wounds related to surgical procedures such as amputation, incision, circumcision, and episiotomy.
Traumatic and pyogenic wounds, vaginitis, cervicitis, pilonidal cyst wounds, and cataract scar tissue.
Preparation of skin graft sites.
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