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Title:
USE OF GLUTEN FOR WOUND TREATMENT AS ACTIVE HEALING INGREDIENT
Document Type and Number:
WIPO Patent Application WO/2007/084127
Kind Code:
A1
Abstract:
A treatment protocol for chronic wounds, such as ulcers, using gluten as the active treatment ingredient. A plant material containing gluten protein is processed to obtain a compound of at least 70% gluten protein. The processed gluten can be applied directly to the wound or combined with a base topical ointment or cream as the active component. The gluten comprises at least 2.5% of the topical ointment or cream by weight, or can be applied directly to the wound as 100% gluten. The gluten is applied at least twice per day. Conventional treatment of rinsing, cleaning, and debriding as required are maintained as well as follow-up monitoring of the patient by a physician at least twice per week until healed.

Inventors:
PETERS, Verdon (P.o.box 397, Floresville, Texas, 78114, US)
OLIVER, Larry (7538 Fair Oaks Parkway, Boerne, Texas, 78015, US)
ANDERSON, Michael (25615 Lewis Ranch Road, New Braunfels, Texas, 78132, US)
Application Number:
US2006/001889
Publication Date:
July 26, 2007
Filing Date:
January 20, 2006
Export Citation:
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Assignee:
PETERS, Verdon (P.o.box 397, Floresville, Texas, 78114, US)
OLIVER, Larry (7538 Fair Oaks Parkway, Boerne, Texas, 78015, US)
ANDERSON, Michael (25615 Lewis Ranch Road, New Braunfels, Texas, 78132, US)
International Classes:
A61K31/74
Download PDF:
Claims:

Attorney's Docket No. PlOOl

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CLAIMS

1. A method for treatment of chronic wounds comprising the steps of: applying a topical treatment material containing processed gluten as the active ingredient to the wound at least twice daily in an amount equal to at least .2cc of processed gluten per square centimeter of wound area, said processed gluten comprised of at least 70% gluten protein.

2. The method for treatment of chronic wounds of Claim 1, further comprising the step of: monitoring the wound by a medical professional for granulation and improvement at least twice per week; and cleaning and rinsing the wound as required.

3. The method for treatment of chronic wounds of Claim 2, further comprising the step of: debriding the wound of necrotic tissue as required.

4. The method for treatment of chronic wounds of Claim 1, further comprising the step of: obtaining gluten protein material from an unprocessed gluten protein source containing at least 56% pure protein, the remainder consisting of no more than 1% fat and 14% moisture.

Attorney's Docket No. PlOOl

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5. The method for treatment of chronic wounds of Claim 1, further comprising the steps in the order of: treating an aqueous slurry of gluten plant material with one or more enzymes comprised of the group of proteases and amylases; filtering the slurry to remove solubilized carbohydrates and treating the insoluble residue with one or more proteases to solubilize the protein components; adjusting the pH with acid; filtering the slurry before drying, centrifuging the slurry before drying, or allowing the reaction mixture to dry without filtering to yield a gluten protein extract water soluble at > 90% at 10g/100ml.

6. The method for treatment of chronic wounds of Claim 4, further comprising the step of: preparing corn gluten hydrolysates according to the following steps in the following order: diluting liquid corn gluten comprised of solids in the 15% to 20% range with water to a solids concentration in the range of approximately 5% to 20%; adjusting the pH to between 6.0 to 8.0; adding amylase in the amount of 0.1 to 1.0% dry basis; cooking the slurry at 280° to 340° F for 3 to 4 minutes; adjusting the cooked slurry pH to between about 4.0 to 0.5;

Attorney's Docket No. PlOOl

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cooling the slurry to 140°F and adding a saccharifying amylase in the amount of .01% to 0.1% dry basis and the slurry maintained at 140°F for 8-16 hours; filtering the slurry and then washing filtrate; reslurrying the filter cake in to between 5% to 20% solids.

7. The method for treatment of chronic wounds of Claim 6, further comprising the step of: preparing corn gluten hydrolysates according to the following additional steps in the following order: adjusting the slurry pH to between 7.5 to 9.0 with Ca(OH) 2 adding a alkaline protease in the amount 0.1% to 1.0% dry basis and maintaining the slurry at 50° to 6O 0 C for 2 to 6 hours or until the pH remains constant; adjusting the slurry pH to between 6.0 to 6.8; filtering out the precipitated Ca 3 (PCW) 2 and insoluble residue; drying the clear filtrate to yield a dry solid product having greater than 70% protein with a nitrogen content in the range of 8% to 16%.

8. The method for treatment of chronic wounds of Claim 1, wherein the treatment material comprises between 1% and 100% processed gluten by weight applied to a wound either in pure form directly or as an ingredient in a base ointment or cream, said base comprising a smooth cream or ointment with a good slip upon

Attorney's Docket No. PlOOl

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skin application having solubilizing and penetration enhancement, and exhibiting colorless and odorless properties and exhibiting good spreading capabilities

9. The method for treatment of chronic wounds of Claim 1, wherein the treatment material comprises at least 2.5% processed gluten by weight as an ingredient in a base cream or ointment, said base comprising a smooth cream or ointment with a good slip upon skin application having solubilizing and penetration enhancement, and exhibiting colorless and odorless properties and exhibiting good spreading capabilities

10. The method for treatment of chronic wounds of Claim 1, wherein the processed gluten protein comprises at least 8% nitrogen.

Attorney's Docket No. PlOOl

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11. A treatment substance for direct application to a wound comprised of: a topical treatment material comprising at least 2.5% processed gluten applied directly to the wound at least twice per day; and said processed gluten comprised of at least 70% gluten protein with a nitrogen content of between 8% and 16%.

12. The treatment substance for direct application to a wound of Claim 11, further comprising: a granulated form of processed gluten, with said processed gluten added to a base topical ointment or cream in an amount to comprise between 2.5% and 75% by weight added to the base topical ointment or cream and between 1 micron and 5 microns in diameter, and applied to the wound.

13. The treatment substance for direct application to a wound of Claim 11, further comprising: a granulated form of processed gluten, with said processed gluten added to a base topical ointment or cream to comprise at least 2.5% gluten protein and between 1 micron and 5 microns in diameter, and applied to the wound in the amount of .2cc of gluten protein per square centimeter of wound size.

Attorney's Docket No. PlOOl

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14. The treatment substance for direct application to a wound of Claim 12, wherein the base topical ointment or cream comprises one of the group of:

Lipoderm™;

Xenaderm™; an emollient cream; vanishing cream; petrolatum; pluronic lecithin organasol; and

Aquaphor™.

15. The treatment substance for direct application to a wound of Claim 13, wherein the base topical ointment or cream comprises one of the group of:

Lipoderm™;

Xenaderm™; an emollient cream; vanishing cream; petrolatum; pluronic lecithin organasol; and

Aquaphor™

Attorney's Docket No. PlOOl

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16. The treatment substance for direct application to a wound of Claim 11, wherein the topical treatment application comprises one or more of the following forming an emollient cream base: polyoxyl 40 stearate; propylene glycol; cetyl alcohol; stearly alcohol; isopropyl myristate; and sodium benzoate.

Attorney's Docket No. PlOOl

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17. A method for direct application of a healing topical substance containing gluten protein as the active ingredient onto a wound comprising the steps of: processing a quantity of gluten protein obtained from a plant source to obtain a processed gluten product comprising at least 70% gluten protein with a nitrogen content of at least 8%; and adding the processed gluten to a base topical cream or ointment to comprise at least 2.5% of a treatment cream or ointment by weight as the active ingredient and having a diameter of between 1 micron and 5 microns.

18. The method for direct application of a healing topical substance onto a wound of Claim 17, wherein the treatment material comprises base ointment, cream, or lotion, said base comprising a smooth cream or ointment with a good slip upon skin application having solubilizing and penetration enhancement, and exhibiting colorless and odorless properties and exhibiting good spreading capabilities

19. The method for direct application of a healing topical substance onto a wound of Claim 17, wherein the treatment material comprises between 1% and 75% gluten by weight of a quantity of a base ointment, cream, or lotion, said base comprising a smooth cream or ointment with a good slip upon skin application having solubilizing and penetration enhancement, and exhibiting colorless and odorless properties and exhibiting good spreading capabilities.

Attorney's Docket No. PlOOl

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20. The method for direct application of a healing topical substance onto a wound of Claim 17; applying treatment cream or ointment at least twice daily to obtain a dosage of at least .2cc processed gluten per square centimeter of wound size.

Description:

USE OF GLUTEN FOR WOUND

TREATMENT AS

ACTIVE HEALING INGREDIENT

Technical Field of the Invention

A method for wound treatment employing gluten as the active healing ingredient, especially for chronic wounds.

BACKGROUND OF THE INVENTION

A wound is a disruption of tissue integrity often associated with trauma. Deep injuries into the muscle tissue, the skeletal system, or the inner organs are defined as complicated wounds and usually require emergency medical treatment. Chronic wounds are classified as chronic when they heal very slowly or fail to heal at all. Generally, wounds that do not heal in a timely fashion (generally 6 to 8 weeks) are classified as chronic wounds.

Chronic wounds often result from other serious health problems such as diabetes, heart disease and circulation disorders. Other contributing factors include poor nutrition and abnormal sustained pressure on skin over bony areas. The most common types of chronic wounds are leg ulcers and diabetic foot ulcers. An ulcer is an open sore, usually associated with circulatory problems. In diabetes, poor blood flow and high blood sugar levels can damage nerves in the foot causing a person not to feel pain as well as inhibit healing in general. A cut that goes unnoticed can then go on to develop into a foot ulcer. Diabetic ulcers usually occur on the foot or leg as a result of poor arterial circulation and nerve damage. Venous stasis ulcers are the most common type of leg ulcer and results from poor venous blood circulation. Pressure ulcers are another type of chronic wound and are usually caused by unrelieved pressure. Patients who are elderly, malnourished, bedridden or paralyzed are more prone to this type of wound because they spend long periods of time in contact with a surface without relief of the pressure. Arterial ulcers usually result from "poor circulation" or "hardening of the arteries," and typically occur on the toes or feet. These types of chronic wounds typically need special treatment to prevent more serious health problems.

Every wound to the human body initiates the body's healing mechanisms that are designed to restore tissue integrity through formation of new structures that more or less match the original. While wound healing is often viewed as a locally regenerative process, it should be

more properly viewed as determined by the overall health of the afflicted person. On this level, wound healing depends on diverse factors such as age, nutrition, medication, immunologic status, metabolic condition, and circulatory status. Compromised circulation is perhaps the greatest single contributory factor for people with chronic wounds that are slow healing or that refuse to heal.

The complex interrelationship between the wound and the patient's overall health becomes especially obvious when wound healing is impaired. Wound healing proceeds in three interrelated dynamic phases regardless of the wound type and the degree of tissue damage. These phases are clinically distinguished as an inflammatory or exudative phase for the detachment of deteriorated tissue and for wound cleansing, a proliferative phase for the development of granulation tissue and a differentiation or regeneration phase for maturation, scar formation and epithelialisation. The three phases can be designated as cleansing phase, granulation phase, and epithelialisation phase.

It is obvious that the best prognosis for successful wound healing is found with smooth, closely abutting incision wounds without substantial tissue loss or presence of foreign bodies in areas with good circulatory characteristics. More difficulty ensues for wound healing in those situations where tissue defects have to be refilled or when pus formation interferes with the direct reassociation of the wound edges. In those situations, the wound surfaces are not closely adjacent and are split away from each other. To close these types of wound, new tissue, the so- called granulation tissue, must be grown. For many types of chronic ulcers, this is the type of chronic wound that develops. Healing in these situations is even more problematic in the event of circulatory issues, and such wounds under these conditions can progress to such a degree that amputation of an affected extremity must be performed.

Current treatment options for chronic wounds are somewhat limited. For pressure ulcers, this can include managing the pressure loads leading to the wound. It can also include nutritional assessment and support, wound care, management of bacterial colonization and infection, compression bandages, medication, surgery (e.g. including amputation) and (more recently) hyperbaric oxygen therapy.

Under current practice, chronic wound care requires careful cleaning and usually dressings. There are many options for dressings, but in general non-adhesive absorbent dressings are best in the early stages when the ulcer is moist and as healing progresses, dry nonadherent dressings may be used. Frequency of dressing is determined by how much ooze and material there is in the ulcer. If there is a lot of pus, daily dressings may be needed. As the ulcer heals and dries, less frequent changes (e.g. weekly) are adequate. Topical antibiotics applied to the ulcer are generally not helpful, because bacteria is always present in ulcers. Unless there is infection spreading beyond the ulcer, antibiotics are generally not considered helpful. Pressure bandaging (e.g. elastic stockings) have been shown to speed healing if properly applied, but great care needs to be taken because of the possibility of further compromising already impaired circulation. The regular use of moisturizers and careful use of steroid creams may help, but steroid creams should not be applied directly to the ulcer. Surgery short of amputation can include skin grafting and removal of dead tissue. Some promise has been shown by using maggots especially grown in a sterile environment for cleaning away dead tissue, avoiding surgerical or very painful debridement.

These various treatment options are not entirely satisfactory and may not significantly reduce healing time. These current procedures are difficult to implement often times, expensive, and still require weeks or month of very slow progress before significant healing occurs. There

is a need for a method and procedure that results in rapid healing without requiring radical and expensive treatment options such as surgery or hyperbaric oxygen therapy.

SUMMARY OF THE INVENTION

The invention provides a wound care product that can be used on all types of human or animal ulcers of the skin. Plant protein hydrolysates, preferably selected from the group consisting of corn gluten hydrolysate, wheat gluten hydrolysate, soy protein hydrolysate, and mixtures thereof, have been found to actively promote healing in patients with chronic wounds. Corn gluten hydrolysate, in the form of dry corn gluten, prepared as disclosed herein has been observed to heal ulcers in six to eight weeks.

The invention uses gluten as a unique active ingredient primarily for treating chronic wounds (e.g. ulcers). The invention uses a topical application comprised of between 100% and 2.5% gluten. Gluten can be applied to the wound either as a pure micro powder paste of about 1 and 5 microns in diameter or is combined with a base topical cream or ointment. Lipoderm™, pluronic F 127, Xenaderm™, Aquaphor™, patrolaneum, and an emollient cream were all used in treatment testing. Preferred treatments include ground 100% gluten powder paste (e.g. 80%- 90% gluten after processing commercially available gluten) for direct application on a wound, a topical ointment of 2.5 g per 100 g (e.g. 2.5% processed gluten) mixed with a Lipoderm™ base to form a topical treatment ointment of 2.5% gluten by weight for application to a wound, and a developed emollient cream made from polyoxyl 40 stearate, propylene glycol, cetyl alcohol, stearly alcohol, isopropyl myristate, sodium benzoate, and comprised of 2.5% gluten by weight. Other topical creams or ointments may be used containing gluten added as the active ingredient to comprise between 2.5% and 75% by weight.

Although the present invention has been exemplified primarily by reference to corn, wheat and soy hydrolysates and corn meal gluten, due to the similarity in the amino acid

compositions of the various plant proteins, it is believed that a wide variety of plant protein hydrolysates would be effective in the practice of the invention, including hydrolysates from other grains, rice, and legumes.

BRIEF DESCRIPTION OF THE DRAWINGS

The objects and features of the invention will become more readily understood from the following detailed description and appended claims when read in conjunction with the accompanying drawings in which like numerals represent like elements and in which:

Fig. 1 is a process chart summarizing the invention.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

The invention utilizes a plant protein hydrolysate, corn gluten, as the active ingredient for various wound care options. Gluten is an amorphous ergastic protein found combined with starch in the endosperm of some cereals, notably wheat, rye, and barley. It is also found in corn, soybeans, rice and legumes, and it constitutes about 80% of the proteins contained in wheat. It is composed of the proteins gliadin and glutenin. Gluten is generally used in the food industry and is responsible for the elasticity of kneaded dough, which allows it to be leavened, as well as the "chewiness" of baked products like bagels. It has been associated with damage to the intestine in some people possibly leading to ulcers, but it has not been traditionally been associated with topical medical treatment options. However, corn starch has been known to yield positive results for babies and elderly patient for several years.

Corn gluten meal is commercially available as a byproduct of corn milling. It is made by drying the liquid gluten stream separated from corn during corn wet milling processing. In the wet milling process of corn, the following fractions are obtained: corn starch, corn oil, defatted corn germ, corn hulls, corn steep liquor, and corn gluten (the protein fraction). Corn gluten is typically separated from the starch stream by centrifugation to yield a thick, yellow slurry of corn gluten containing 15% to 20% solids. Conventionally, the corn gluten is filtered and dried to produce solid corn gluten meal, which is generally sold as an animal feed product. This is the source of the gluten experimentally used leading to the claimed invention.

Gluten was used in several experimental medical case studies to determine the efficacy of using gluten as the active ingredient for treating chronic wounds in patients. Granulated 60% corn gluten meal was used manufactured by Grain Processing Corporation (GPC). Table 1 provides GPCs manufacturing specification for the product.

TABLE l: Corn Gluten Specification

The present plant protein hydrolysates are preferably prepared by a process comprising treating an aqueous slurry of a plant protein such as corn or wheat gluten or soy protein with acid or with one or more enzymes. In the process used in developing the invention, the base material was the 60% corn gluten meal. Preferably, the plant protein is treated with one or more proteases, and most preferably, is pre-treated with one or more amylases. For example, the proteinaceous slurry may be treated with amylases, followed by filtration to remove the solubilized carbohydrates. The insoluble residue is then treated with one or more proteases to solubilize the protein components. After pH adjustment with acid, the slurry is filtered and/or centrifuged. The effluent is dried in a conventional manner to yield "corn gluten hydrolysate", "wheat gluten hydrolysate", or "soy protein hydrolysate", which is essentially water soluble (>90% at 10 g/100ml). Alternatively, the protein slurry can be treated with proteases alone and the entire reaction mixture dried, or the reaction mixture may be centrifuged or filtered and the supernatant or filtrate dried in an appropriate manner, to yield a soluble plant protein hydrolysate.

To prepare corn gluten hydrolysates, the liquid corn gluten (15%-20% solids) is preferably diluted with water to a solids concentration in the range of approximately 5% to 20% and the pH adjusted to about 6.0 to 8.0, preferably to about pH 6.5. The appropriate amylase is added (0.1 to 1.0% dry basis (DB)) and the slurry jet cooked at 280° to 340° F, preferably at 320° F for 3-4 minutes. The cooked slurry is then adjusted to about pH 4.0 to 0.5, cooled to

140° F, and optimally, a saccharifying amylases (glucoamylase) is added (.01% to 0.1% DB) and the slurry maintained at 140 0 F for 8-16 hours, preferably 12 hours.

The slurry is then filtered and washed and the filtrate and washings discarded. The filter cake is reslurried in water to 5% to 20% solids (preferably approximately 10%) and adjusted to pH 7.5 to 9.0 with Ca(OH) 2 .. An alkaline protease is then added (0.1% to 1.0% DB) and the slurry is maintained at 50° to 60°C for 2 to 6 hours, or until the pH remains constant. The slurry is then adjusted to pH 6.0 to 6.8 (preferably pH 6.2), and the precipitated Ca 3 (PO 4 ) 2 and insoluble residue is removed by filtration. The clear filtrate is then dried in an appropriate manner (i.e. spray drying, drum drying, etc.) to yield a dry solid product having greater than about 80%-90% protein (Kjeldahl nitrogren), and which is essentially water-soluble at 10 wt~% concentration. On a dry basis, the corn gluten hydrolysate will have a nitrogen content of at least about 8%, in the range of 8%-16%, and most preferably at about 14.4%. The gluten was finely reduced to a ground powder form of approximately 1 to 5 microns in diameter.

Four different gluten types were prepared and used to develop the invention as follows:

1) "GLUTEN #1": Pure plant form that is a filtered base and then freeze-dried.

2) "GLUTEN #2": Comprised of soluble and insoluble filtrates approximately 69% protein.

3) "GLUTEN #3": Appropriately filtrated to increase the protein purity.

4) "GLUTEN #4": The most water soluble and pure form is greater than 90% at 10g/100ml.

After preparation, the gluten was applied to wounds as a topical active treatment using several different techniques. This included direct application of the prepared gluten to the wound and mixing with several different therapeutic base preparations that included Lipoderm™, emollient cream, vanishing cream, petrolatum, pluronic lecithin organasol, Aquaphor. The case studies using this treatment options have demonstrated that the gluten protein possesses a bacteriostatic and bacteriological effect as well as acting as a healing stimulant on open ulcers and other

wounds. The active component is the protein component of the corn gluten meal, but it is believed that any source of gluten prepared as above will provide the same treatment effectiveness.

Generally, the composition of the unprocessed gluten is 15% gluten protein, 70% starch, and the remaining 15% salts, polypeptides, and various amino acids. After processing, the gluten composition comprised at least 70% protein and generally should consist of 80% to 90% gluten protein with the remaining additional 10% to 30% comprising varying amounts of amino acids, salts, polypeptides, and starch. The ratio for medical application is 0.2cc of gluten to 1.0 cm of wound surface. This can be applied twice daily or as directed by the treating physician. Treatment application varies and is not to exceed two to three days without reapplication. The most active form of gluten observed was gluten #2 in a Lipoderm™ transdermal base.

Medically, when this processed form of gluten is topically applied to an ulcer, the ulcer responds with granulation and rapid decrease in odor and drainage. The fibrin character of a chronic ulcer is also observed to rapidly diminish as well. This positive cellular synergistic stimulation decreases colonization of bacteria until the granulation reaches the surface (skin height). At that point, epithelization begins at the ulcer margins over the next several weeks as the ulcer continues to close and heal. This healing process has been observed to occur over a period of approximately 4 to 8 weeks, depending on the size of the ulcer and other systemic conditions of the patient. Typical bases (dispersant vehicles for the gluten) included or can include Lipoderm™, emollient cream, vanishing cream, petrolatum, pluronic lecithin organasol (e.g. pluronic F 127), Xenaderm™, and Aquaphor™. Lipoderm™ was used as the base ointment that demonstrated the best results.

Lipoderm™ is a brand item comprised of a liposomal component that provides a high chemical penetration enhancer value, allowing medication to reach the circulatory system and

local tissue quickly. Xenaderm™ is a brand item comprised of Trypsin 90 units, Balsam Peru 87mg, Castor Oil 788 mg, and inactive ingredients Safflower Oil, and aluminum magnesium hydroxide stearate. It is an effective capillary bed stimulant that increases circulation in the wound site area. Pluronic F 127 is a polaxamer emulsifier with soap-like characteristics that performs as a carrier of water-soluble drugs for percutaneous absorption of active medical ingredients. It is odorless, colorless, and non-greasy. When applied to the skin, it thickens rapidly before penetrating and drying. Aquaphor™ is a petrolatum base that is widely used as a dermatological base. It is a neutral anhydrous base for compounding stable emulsifers, highly miscible with both aqueous solutions and oil based substances. Petrolatum is a colorless to yellowish white hydrocarbon mixture obtained by fractional distillation of petroleum. It is used in preparations of medicinal ointments and lubrication. AU of these inactive base ointments are a smooth cream with a good slip upon skin application having solubilizing and penetration enhancement properties. They are essentially colorless, odorless, and exhibit good spreading capabilities. Adjunctive therapies used along with the gluten treatment included Metronidazole, Gentamycin, Mupricion, Papain Urea, Silver Sulfur compounds, Clotrimazole, Copper complex sodium, Diflucan, Nystatin, Triamcinolone, Nifedipine, and Pentoxifylline.

Approximately 40 patients were treated in informal medical trials involving gluten as a topical treatment option. Five exemplary case studies are presented as follows showing the effectiveness of gluten. CASE STUDY #1

The first case study involved a patient with an ulcer on left ankle and foot of approximately two years duration and with a history of extremely poor hygiene and unhealthy living conditions. The patient was first examined on November 15, 2002 suffering from an ulcer. Over the course of the next two years, the patient received standard chronic wound care

treatment that included cleaning of the effected area, spraying with .25% acetic acid, applying various dressings, debriding necrotic tissue, rinsing with .09% sterile normal saline, treating with antifungal cream, and treating with antibiotics between November 15, 2002 and March 30, 2004.

On March 30, 2004, the patient received the first treatment of gluten applied directly to the wound. Positive results were noted by April 2, 2004. On April 9, 2004, it was noted that wound was showing granulation, and after removal of maggots on April 14, 2004 excellent granulation was noted and gluten was applied. During the period April 2, 2004 to September 24, 2004 granulation and hearing was noted along with improvement in appearance to complete healing. Both 100% gluten was applied to the wound and a 50/50 mix of gluten and silver sulfadiazine were used in the treatment regimen in addition to traditional cleaning and rinsing.

On June 1, 2004, it was noted that the ulcer was not exhibiting an additional healing activity and treatment of the 50/50 mix was discontinued in favor of the 100% gluten application. Increase granulation was again present on June 4, 2004, indicating better results with 100% gluten compared to the 50/50 mix. By June 15, 2004, it was noted that skin was rapidly bridging the ulcers. Both ulcers were noted as essentially healed except for a small area on August 10, 2004. Total closure of the ulcers was noted on August 17, 2004, with both ulcers completely healed and the patient discharged on September 24, 2004. CASE STUDY #2

The patient presented on July 20, 2004 with ulcers on both feet. Conventional treatment consisted of cleaning with techncare, debriding dead tissue using jetox, and treating with Xenaderm™ cream containing 2.5% gluten. Ulcer sizes and locations were noted on August 13, 2004 as being a 20mmx20mmxlmm ulcer on left shin, a scabbed 15mmx5mm wound on the left foot, and a scabbed 20mmxl0mm wound on the right foot. The ulcers initially showed slow

steady improvement over the course of the several weeks of treatment and were completely closed and healed by August 20, 2004. CASE STUDY #3

A 57-year old diabetic female presented August 9, 2004 with a Wagner grade IV ulcer of 15mrnx30mrnxl 9mm on her lower back region. She was medically evaluated and started on hyperbaric treatment (2.0 ATA, 100% oxygen x 90 minutes). Treatment also included application of accuzyne and dressings. This treatment protocol was used approximately thirty times. The wound showed good response to treatment to the 20 th dive, however, little wound healing was evident until Gluten was started on August 31, 2004. As of August 22, 2004, Hyperbaric Oxygen therapy was discontinued and the patient was converted to wound care only with Gluten. The conventional treatments resulted in some improvement to the wound including granulation, but significant healing progress did not occur. She was seen three times per week from August 31, 2005 and treated with Gluten, and as of October 18, 2004, the wound was totally healed and she was discharged. CASE STUDY #4

A 60-year Insulin dependent diabetic male presented with a three-week history of infection of his transmetatarsal amputation. He was referred by his family physician on July 23, 2004 for evaluation of hyperbaric oxygen therapy and wound care management. The patient was started on hyperbaric treatment (2.0 ATA, 100% oxygen x 90 minutes) on July 26, 2004, and wound care at this time was started using Gluten #2. By July 28, 2004, rapid granulation of the wound was noted, and conventional therapy continued with cleaning and debridement of necrotic tissue as well as gluten and hyperbaric treatment.

The treatment regimen was switched to Gluten #1 on August 23, 2004 by treating the upper half of wound with that gluten preparation and treating the lower half with flagyl-triple

(50/50) along with conventional debridement. On August 26, 2004, the size of the wound was observed to be 30rnmx30mm with continued positive response, and the treatment was switched to 100% gluten powder on the wound. By September 2, 2004, the wound size was recorded as 23mmx30mm with granulation still improving.

The wound showed rapid improvement in response to the gluten powder applied directly to the wound, and by September 9, 2004 the wound had shrunk to 20mmx30mm with very good granulation. Rapid healing was noted from September 2, 2004 to September 23, 2004 with wound size reduced to 13mmxl8mm by September 23, 2004. By October 7, 2004, the wound size was 10mmxl4mm, and on October 8, 2004 it was noted that epithelization of the edge of the wound had increased. Between October 15 and October 22, 2004, the wound closure had improved by 50%. The patient progressed slowly but with positive progress with the wound, and on October 29, 2004 hyperbaric treatment was discontinued after sixty treatments. Patient was placed on conventional wound care using gluten alone. The ulcer showed complete closure on December 27, 2004 when the patient was discharged. CASE STUDY #5

The patient first presented on May 6, 2004 suffering from an ulcer on the ankle. The ulcer slowly improved with conventional treatment options utilized. The first treatment of gluten was applied on May 26, 2004, and by June 2, 2004 the ulcer measured 20mmx20mmx3mm in size with positive healing progress. On June 9, 2004, granulation was noted with slow improvement of the ulcer and further treatment using gluten. On June 9, 2004, the ulcer measured 20mmx25mmx.5mm in size, and the ulcer was treated with conventional rinsing and cleaning along with gluten.

On July 9, 2004, the ankle was noted as having inflamed cellulites and swollen with edema but there was no drainage. Gluten 2.5% with a combination of mycolig nystatine and

bactroban in Aquaphor™ (e.g. referred to hereafter as "Gluten triple") was applied. By July 14, 2004, the right ankle was noted as dry and healing, while left foot exhibited a partially detached toenail. Gluten triple was applied along with a dressing. The size of the ulcer on the ankle was 20mmx20mmx2rnrn by July 21, 2004. The ulcer continued to be treated using Gluten 2.5% with a combination of mycolig nystatine and bactroban in Aquaphor, and on July 28, 2004 the size of the ulcer was noted as 17rnmxl7mmx3mm.

On August 11, 2004, the ulcer was noted as clean with decreased erythema and no drainage. The size of the ulcer was noted as 17mmxl7rnmx2mm, and the ulcer was cleaned and rinsed and treated with Gluten #2. The patient exhibited a history of failing to wear a protective pad as directed, and by September 15, 2004, it was noted that the size of the ulcer had increased to 18rnmxl8mmx2mrn. Gluten #2 was applied, and by October 20, 2004, progress had been noted and the size of the wound had substantially reduced to 15mmxl3mm. On November 10, 2004, the ulcer was debrided and measured 6mmx4mmx2mm. By November 17, the ulcer was almost healed with a good scab cover, and on December 8, 2004, the ulcer was healed. OTHER CASE STUDIES SUMMARY

A total of about 40 case studies were performed using gluten as the active component of a topical treatment regimen that included conventional treatment options. The results of the various treatments showed positive signs of healing in the form of granulation within a week of the first treatment of gluten followed rapidly by noticeable epithelialisation of the tissue and shrinkage of the wound. This response was consistent even among patients with ulcers of long duration that had proven very resistant and non-responsive to other conventional treatments, such as found in Case Study #1. Wound healing progressed from within four weeks to as long as six months. In most patients, wound healing was essentially complete within eight weeks from beginning the gluten treatment.

TREATMENT PROTOCOL, INGREDIENTS, AND FORMULATION

The gluten used in the case studies was granulated 60% corn gluten meal manufactured by Grain Processing Corporation (GPC) and processed as stated above. Corn gluten is a commercially available material extracted from meal by drying the liquid gluten stream separated from corn during wet milling processing. While the specific composition can vary, generally commercially available gluten consists of about 60% protein with the remainder made up lipid, carbohydrate, and ash material. The following table sets forth the typical protein composition of corn gluten in this commercial form:

TABLE 2: Corn Gluten Protein Content

The processed gluten product used in the testing consisted of micro powdered gluten milled to a size of between 1 to 5 microns in diameter. The processed gluten was comprised of at least 70% gluten protein, generally in the range of 80% to 90% protein and containing about 12% nitrogen. The gluten was then mixed with glycerin to form a paste and generally added to a base topical preparation in the form of a cream or ointment. However, the pure gluten protein

paste was also used in testing. The following tables set forth additional information on the gluten formulations.

TABLE 3: Gluten Micro Powder Composition

TABLE 4: Lipoderm™ Base + Gluten Product

TABLE 5: Xenaderm™ Base + Gluten Product

TABLE 6: Pluronic F127 Base + Gluten Product

TABLE 7: Aquaphor™ Base + Gluten Product

Recommended Suggested Compounding Procedures

1. At a temperature of 80°-85°C, mix all the ingredients of Step 1 through 3. Remove from heat and continue mixing until temperature drops to 45°C.

2. Dissolve the Sodium Benzoate in 5ml purified water, USP and disperse into the cream. Let age for one or two days.

3. Weigh, and using a mixer, add enough purified water, USP to make 450grams.

4. Add at least 11.25 grams of gluten micro powder to form a topical cream containing at least 2.5 grams per 100 grams of cream. Use mixer to thoroughly mix and disperse the powder throughout the mixture.

Table 8: Suggested Formula for Emollient Cream

Treatment with 2.5% gluten powder in Lipoderm™ base demonstrated the best observed healing response. Rapid granulation and healing began within one week of the first application and provided positive visual results upon the next visit after the first application. An active base amount of gluten less than 1.0% demonstrated little or no improvement to a wound after treatment and was not regarded as an effective treatment amount. Gluten as a percent of total amount of application that obtained noticeable and desired healing results ranged from greater than 1% to 100%. These best results were also obtained from an application of a gluten topical treatment of at least 2.5% applied twice daily in an amount sufficient to cover the entire wound. Optimal results were obtained for a topical treatment concentration equaling .2cc processed

gluten protein per 1.0 cm 2 wound surface area applied twice daily and not exceeding two to three days without reapplication. It is important that continued application under this protocol occur until a scab forms over the wound, indicating continued healing with adequate circulation and clearing of active bacterial infection.

The gluten can be used alone or as a component of a treatment cream or ointment, with a composition of 1% to 100% by weight, preferably in the range of 2.5% to 75% as an ingredient to a cream or ointment base. Although Lipoderm™ is preferred, any base topical ointment is acceptable. Topical ointment bases used in the case studies showing acceptable wound healing included Xenaderm™, petrolatum, Pluronic F 127, Aquaphor™, and the emollient cream described above, but it is believed that any topical base with similar characteristics provides acceptable results as long as a gluten micro powder of at least 70% gluten protein in the size of 1 to 5 microns is present to form a >2.5% gluten treatment composition ointment.

The preferred treatment procedure is to apply the gluten treatment composition to the wound at least twice daily along with other conventional treatment protocols as required. For example, debridement of necrotic tissue, once or twice a week, should be continued, with care exercised not to remove granulation tissue. Rinsing and cleaning of the wound as required should also be performed. Patients should also use prescribed protective covers, but the gluten application does not require that any protective covering be used such as a dressing. The prescribed protective covering is primarily needed to prevent the topical gluten treatment from being wiped away from the wound and to protect the wound from painful contact with clothing while it heals. The application protocol of at least .2cc gluten per 1.0 cm wound area applied at least twice daily is the preferred treatment protocol. Examination by a treating physician or other medical professional should also be performed at least twice weekly to monitor healing and perform other treatments, such as debridement of necrotic tissue, when required.

Figure 1 is a process flow chart summarizing the invention. In step 5, gluten protein is obtained from a plant source, such as corn, wheat, rye, barley, soybeans, rice, or legumes. The gluten protein is processed in step 10 to obtain a gluten product comprising at least 70% gluten protein in granulated powdered form. This gluten protein should also comprise at least 8%, generally between 8% and 16%, nitrogen. The powdered gluten is mixed with glycerin to form a paste in step 15. In step 20, the gluten paste is mixed with a topical base ointment or cream or left in pure form, hi step 25, the gluten topical treatment product is milled to ensure a gluten treatment product of beween 1 and 5 microns in diameter. In step 30, the gluten treatment product is applied at least twice daily to a wound in sufficient quantity to equal at least .2cc of gluten protein per centimeter of wound surface area. Preferrably, the gluten topical treatment will comrpise between 2.5% and 75% gluten protein by weight.

While the invention has been particularly shown and described with respect to preferred embodiments, it will be readily understood that minor changes in the details of the invention may be made without departing from the spirit of the invention. Having described the invention, we claim: