Pharmaceutical compositions comprising modified fucoidan for the treatment of fibrous adhesions and other diseasesThe application is a divisional application, the application date of the original application is 2010, 7 months and 27 days, the application number is 201080042840.4, and the invention name is 'the pharmaceutical composition containing the modified fucoidin for treating fibrous adhesion and other diseases'.
CROSS-REFERENCE TO RELATED APPLICATIONS
This application claims priority from U.S. provisional patent application No.61/228,929 filed on 27/7/2009. This application and all other documents listed herein are incorporated by reference into this specification.
Directory
The following is a list to aid in the review of the present application.
CROSS-REFERENCE TO RELATED APPLICATIONS
Background
Disclosure of Invention
Detailed Description
General discussion of exemplary drugs
Fucoidin (fucan)
Film
Gel
Drip agent
Discussion of quantitative efficacy of anti-fibrous adhesion Agents
Other exemplary embodiments
Examples
Claims
Abstract
Background
A fibrous adhesion is a scar that typically forms between two parts of the body after surgery (surgical adhesion). Fibrous blocking can cause serious problems. For example, fibrous adhesions involving the female reproductive organs (ovaries and fallopian tubes) can cause infertility, dyspareunia, and severe pelvic pain. Fibrous adhesions that occur in the intestine can cause ileus or obstruction, and can also form elsewhere, such as around the heart and spine, and the hands. In addition to surgery, fibrous adhesions may also be caused, for example, by endometriosis, infection, chemotherapy, radiation, trauma, and cancer.
Various fibrous adhesions are discussed in this document. Terms such as surgical adhesions, post-surgical adhesions, adhesions due to pelvic inflammatory disease, adhesions due to mechanical injury, adhesions due to radiation therapy, adhesions due to trauma, and adhesions due to the presence of foreign bodies all refer to the adhesion of tissues to one another by similar mechanisms and are all included in the term fibrous adhesions.
Fibrous adhesion formation is a complex process in which tissues that are normally separated in the body grow together. Surgical adhesions (also known as post-surgical adhesions) are formed by an otherwise normal wound healing response of tissue to trauma and have been reported to occur in more than two thirds of all abdominal surgical patients (Ellis, h., surg. gynecol. obstet.133:497 (1971)). The results of these fibrous adhesions vary and depend on the surgical site or other sites involved (e.g., the site of disease). Problems may include chronic pain, ileus and even an increased risk of death following cardiac surgery (DiZerega, G.S., prog.Clin.biol.Res.381:1-18 (1993); DiZerega, G.S., Fertil.Steril.61:219-235 (1994); Dobell, A.R., Jain, A.K., Ann.Thorac.Surg.37:273-278 (1984)). Fibrous adhesions involving the uterus, fallopian tubes or ovaries are estimated to account for about 20% of all infertility cases in women of childbearing age (Holtz, G., Fertil. Steril.41: 497-353 (1984); Weibel, M.A.and Majno, G.Am.J.Surg.126:345-353 (1973)).
The process of fibrous adhesion formation initially involves the construction of a fibrin framework and normal tissue repair. The normal repair process allows fibrinolysis and mesothelial repair. However, during the formation of fibrous adhesions, the fibrin matrix matures as fibroblasts proliferate into a network and angiogenesis occurs, resulting in the formation of organized fibrous adhesions within about 3-5 days (Buckman, R.F., et al., J.Surg.Res.21:67-76 (1976); handoff, A.T., J.Ant.129: 659-664 (1979)). Inflammatory processes include neutrophil activation in the injured tissue, fibrin deposition and adhesion of adjacent tissues, macrophage invasion, fibroblast proliferation into areas, collagen deposition, angiogenesis and formation of permanent fibrous adhesion tissue.
Various attempts have been made to prevent surgical adhesions. These include pharmacological approaches directed at affecting biochemical and cellular activities that occur with trauma and barrier approaches to isolating the affected tissue. For example, all methods of using peritoneal lavage, heparinized solutions and procoagulants, improving surgical techniques (e.g., using microscopic or laparoscopic surgical techniques), removing talc from surgical gloves, using smaller sutures and using physical barriers (films, gels or solutions) to minimize apposition of serosal surfaces have been attempted. At present, prophylactic therapy also includes the prevention of fibrin deposition, the reduction of inflammation (steroidal and non-steroidal anti-inflammatory drugs) and the removal of fibrin deposits.
Interventional attempts to prevent post-surgical adhesion formation have included the use of hydrated floatation techniques or barrier devices. The hydroplaning technique involves instillation of large volumes of polymer solution, such as dextran (Adhesion study Group, Fertil Stel.40: 612-. Regeneration of cellulose by oxidation (e.g. interceded)TM) And polytetrafluoroethylene (Gore-tex surgical membrane) and modified hyaluronic acid/carboxymethylcellulose (HA/CMC) composition (Seprafilm)TM) Manufactured fully absorbable membranes have also been used in animals and humans to reduce post-surgical adhesion formation (Burns, j.w., et al, eur.j.surg.suppl.577:40-48 (1997); burns, J.W., et al, Fertil. Steril.66:814-821 (1996); becker, J.M., et al, J.Am.Coll.Surg.183: 297-. The success of these HA/CMC membranes derives from their ability to provide tissue separation when fibrous adhesions form during peritoneal wound repair. The films were observed to form a clear, tacky coating on the damaged tissue for 3-5 days after application, which is consistent with the time course of adhesion formation after surgery (Ellis, h., br.j.surg).50:10-16(1963)). Unfortunately, only limited success has been seen with these approaches.
Peritonitis involves inflammation of the peritoneum. Peritonitis can cause serious problems. For example, abdominal pain, abdominal tenderness, and abdominal strain. Peritonitis may include spontaneous, anatomical, and/or peritoneal dialysis-related inflammation. Peritonitis may involve infections such as perforation of the hollow organs, peritoneal rupture, and idiopathic bacterial peritonitis, and systemic infections may lead to infection and peritonitis. Peritonitis may also involve no infection, e.g., leakage of sterile body fluids into the peritoneum, and sterile abdominal surgery may result in peritonitis. Various attempts have been made to prevent and/or treat peritonitis. For example, general supportive measures (such as intravenous fluid supplementation), antibiotics, and surgery. There is an unmet need for compounds, compositions, methods, etc. (including methods of delivery) that inhibit or treat and/or prevent peritonitis, preferably more effectively with little or no side effects.
Ischemia or ischemia involves a limitation in blood supply, which may include an inadequate supply of oxygen, glucose, and other components required for proper tissue function, resulting in tissue damage and/or dysfunction. Ischemia can cause serious problems. For example, the tissue may become hypoxic and necrotic, and clots may form. Various attempts have been made to prevent and/or treat ischemia. For example, blood flow restoration or reperfusion. However, blood recovery involves reintroduction of oxygen, which may cause additional damage due to the generation of free radicals, resulting in reperfusion injury. Reperfusion injury can cause serious problems. There is an unmet need for compounds, compositions, methods, etc. (including methods of delivery) that inhibit or treat and/or prevent ischemia and/or reperfusion injury, preferably more effectively with little or no side effects.
Endotoxemia is the presence of endotoxin in the blood. Endotoxemia can cause serious problems. For example, endotoxemia can lead to septic shock. There is an unmet need for compounds, compositions, methods, etc. (including methods of delivery) that preferably inhibit or treat and/or prevent endotoxemia more effectively with little or no side effects.
Keloid features cause the wound to heal with raised scars. Raised scars, characteristic of keloids, are involved in abnormal fibrous scarring. Keloid features cause serious problems. Such as pain and disfigurement. There is an unmet need for compounds, compositions, methods, etc. (including methods of delivery) that inhibit or treat and/or prevent the characteristics of keloid and the raised scars that result therefrom, preferably more effectively with little or no side effects.
Keloid (keloid) is a scar that expands during growth on top of normal skin. Keloids involve abnormal collagen growth, including abnormal growth of type I and type iii collagen. Keloids cause serious problems. Keloids, for example, cause pain and itching, and ulcers may occur if infected. Various attempts have been made to treat or prevent keloids, including the use of surgery, dressings, steroid injections, and laser therapy. There is an unmet need for compounds, compositions, methods, etc. (including methods of delivery) that inhibit or treat and/or prevent keloids, preferably more effectively with little or no side effects.
Dermatitis includes inflammation of the skin, including atopic dermatitis and contact dermatitis. For example, contact dermatitis involves localized rashes and/or skin irritation after the skin has been in contact with foreign matter. For example, atopic dermatitis is a chronic recurrent pruritic skin disease. Atopic dermatitis is sometimes referred to as beneiezatid, neurodermatitis, endogenous eczema, eczema flexor, infantile eczema, childhood eczema, and prurigo diathsique. Eczema is a form of dermatitis. Other types of dermatitis include acanthocytic edema dermatitis, seborrheic dermatitis (dandruff), dyshidrotic dermatitis (pompholyx), urticaria, vesicular dermatitis (bullous dermatitis), and papular urticaria (urticaria). Dermatitis can cause serious problems. For example, dry skin, rashes, skin edema, redness, itching, crusting, cracking, blistering, weeping, and bleeding. Various attempts have been made to treat or prevent dermatitis, including the use of corticosteroids and coal tar. There is an unmet need for compounds, compositions, methods and the like, including methods of delivery, that preferably inhibit or treat and/or prevent dermatitis (including atopic dermatitis, eczema, contact dermatitis, acanthotic edema dermatitis, seborrheic dermatitis, dyshidrotic dermatitis, urticaria, vesicular dermatitis, and papular urticaria) more effectively with little or no side effects.
Erythema acne is a chronic disease or condition typically characterized by facial erythema. Rosacea may cause serious problems. For example, rosacea typically starts with redness of the forehead, nose or cheeks, and can also cause redness of the neck, ears, scalp and chest. For example, rosacea may cause other symptoms including telangiectasia, papules, pustules, and painful sensations, and in advanced cases may develop hypertrophic rosacea (red lobular nose). The erythema and acne subtypes include erythema and angiectasis type erythema and acne, papulopustule type erythema and acne, wart type erythema and acne and eye erythema and acne. Various attempts have been made to treat or prevent erythema acne, including the use of anti-inflammatory drugs and antibiotics. There is an unmet need for compounds, compositions, methods, etc. (including methods of delivery) that preferably more effectively inhibit or treat and/or prevent rosacea (including its erythematoid, papulopustular, excrescent, and ocular subtypes) with little or no side effects.
There is an unmet need for compounds, compositions, methods, etc. (including methods of delivery) that preferably inhibit or treat and/or prevent fibrous adhesion formation and/or other diseases and/or conditions discussed in this specification and related diseases and/or conditions more effectively with few side effects. The compounds, compositions, methods, and the like of the present invention provide one or more of the foregoing advantages.
Disclosure of Invention
The compositions and methods, etc. of the invention in this specification include compositions and methods, etc. comprising one or more agents against fibrous adhesions or other diseases discussed in this specification, which are useful for treating surgical adhesions or other such diseases or conditions. Anti-disease drugs provide significant therapeutic effects on fibrous adhesions or other conditions while also generally providing low side effects. In addition, because a plurality of different drugs are discussed, combinations of drugs can be selected as needed to reduce side effects in patients who may suffer from other diseases or conditions, and/or to provide other beneficial health effects or therapeutic effects, such as a composition that not only inhibits fibrous adhesions, but also treats any of cancer or arthritis or swelling or a variety of other diseases or conditions (which may also be treated with one or more anti-fibrous adhesion agents in this specification). The compositions of the present specification can also be used to treat conditions characterized by fibrous growth and, for example, keloids that have similar biology as fibrous adhesions, as well as other diseases and conditions discussed in the present specification. Therefore, the discussion in this specification is equally applicable to such fiber growth.
In some embodiments, the compositions and methods, etc., of the present invention include in the present specification treatment, prevention, inhibition, etc., using selected modified fucoidan (or fucoidan composition) comprising a specific combination of components that generally includes: a total carbohydrate content of more than about 40% w/w; a trehalose content of about 40-100% by weight of the total carbohydrate content; a galactose content of about 0-60% as a percentage of the total carbohydrate content; a sugar content excluding (i.e., excluding) trehalose and galactose of about 0-20% as a percentage of total carbohydrate content; an acetyl group to trehalose monomer ratio of less than about 40%; a molecular weight distribution such that about 0 to 5,000g/mol of the modified fucoidan moiety comprises less than about 30% w/w; such that about 5,000-60,000 g/mol of the modified fucoidan moiety comprises a molecular weight distribution of less than about 50% w/w; such that about 60,000 to 200,000g/mol of the modified fucoidan fraction represents less than about 40% w/w of the molecular weight distribution; such that about 200,000 to 1,600,000g/mol of the modified fucoidan moiety comprises a molecular weight distribution of less than about 50% w/w; such that greater than about 1,600,000g/mol of the modified fucoidan moiety comprises a molecular weight distribution of less than about 50% w/w; a sulfate content of about 10-50% w/w; and/or a water content of less than about 20% w/w. The selected modified fucoidan also includes modified fucoidan that forms a solution having a pH of about 4 to about 8 when prepared as a 0.1% w/v solution. The compositions typically comprise at least one pharmaceutically acceptable excipient, filler, carrier or diluent. If desired, the pharmaceutically acceptable excipient, filler, carrier or diluent may be selected from the group consisting of pluronic, cellulose, alginate, acrylate, hyaluronic acid, polyethylene glycol and chitosan.
Thus, the compositions and methods, etc., in the present specification include modified fucoidan for the treatment of inflammatory diseases, including arthritis, for the treatment of fibrous adhesions, including surgical adhesions, and for the treatment of peritonitis, ischemia, reperfusion injury, endotoxemia, keloid-characterized scarring, keloids, dermatitis, and rosacea.
Thus, the compositions and methods, etc., of the present invention provide, in the present specification, pharmaceutical compositions formulated to inhibit fibrous adhesions, the compositions comprising a therapeutically effective amount of a modified fucoidan selected for inhibiting fibrous adhesions as described in the present specification, optionally at least one therapeutically effective agent selected for inhibiting fibrous adhesions in the present specification, and at least one pharmaceutically acceptable excipient, filler, carrier, or diluent. If desired, the pharmaceutically acceptable excipient, filler, carrier or diluent may be selected from the group consisting of pluronic, cellulose, alginate, acrylate, hyaluronic acid, polyethylene glycol and chitosan.
The compositions and methods of the invention, among other things, comprise selected modified fucoidans for treating peritonitis, ischemia, reperfusion injury, endotoxemia, keloid-characterized scarring, keloids, dermatitis, and rosacea. The modified fucoidan provides significant therapeutic effects on peritonitis, ischemia, reperfusion injury, endotoxemia, keloid-characterized scarring, keloid, dermatitis, and rosacea, while generally providing low side effects. In one aspect, the compositions and the like provide a method for treating or preventing peritonitis, ischemia, reperfusion injury, endotoxemia, keloid-characterized scarring, keloids, dermatitis, and rosacea in a subject or patient comprising administering to the subject or patient a therapeutically effective amount of the selected modified fucoidan.
In some embodiments, the subject or patient is an animal, such as a human, dog, cat, horse, cow, camel, or other mammal, or a bird, reptile, or other animal. The site may be an animal as a whole, or a local site, such as an intra-abdominal, limb, spinal column, head, reproductive tract, gastrointestinal tract, pulmonary system, thoracic cavity, heart or vascular system and urinary system, or an intra-abdominal site for the treatment of peritonitis, ischemia and reperfusion injury, or a vascular system for the treatment of endotoxemia, or a local site for the treatment of a disease characterized by keloid scarring, keloids, dermatitis and rosacea, or any other desired site. The site may be an animal as a whole, or a specific site within the abdomen, limb, spine, head, reproductive tract, gastrointestinal tract, pulmonary system, thoracic cavity, cardiac or vascular system, urinary system, on the skin, or any other system or location as desired. The treatment site may be a surgical site, a pelvic inflammatory site, a mechanical injury site, a radiation exposure site, a site damaged by the presence of foreign matter, or any other desired site.
The selected modified fucoidan can be administered substantially continuously to the disease site by controlled release from the polymeric dosage form. Administration forms may include films, instillations, patches, pastes, microspheres, implants, gels, sprays or liquids, solutions and suspensions, which may be in lactated ringer's injection usp (lactated Ringers injectionusp). The selected modified fucoidan can be administered in combination with a second agent, which can be any one or more of the other agents of the present specification or any other therapeutic agent.
The compositions and methods, and the like, can be used to prepare a medicament, including a pharmaceutical composition or device, for alleviating symptoms associated with fibrous adhesions, peritonitis, ischemia, reperfusion injury, endotoxemia, keloid-characterized scarring, keloids, dermatitis, and rosacea in a subject or patient. The medicament may be prepared by combining a pharmaceutically effective amount of the modified fucoidan and a pharmaceutically acceptable excipient, diluent, or buffer, and may comprise an additional anti-disease agent.
The modified fucoidan compositions of the present disclosure can be used, for example, in combination with a therapeutically effective amount of one or more second therapeutic agents selected from the group consisting of: alginic acid, doxycycline, cortisone, estramustine, melezitose, succinic acid, meclofenamate sodium (meclofenamate), palmitic acid, dextran sulfate, collagen, budesonide, enalapril (e.g., enalapril maleate), nabumetone, statin (e.g., simvastatin), captopril, chitosan, minocycline, methotrexate, cisplatin, ibuprofen, erythromycin, tetracycline, an SDF-1 inhibitor (e.g., anti-SDF-1 antisense oligonucleotide (ASO), anti-SDF-1 small molecule RNA, anti-SDF-1 siRNA, anti-SDF-1 ribozyme, anti-SDF-1 aptamer, a small molecule inhibitor of SDF-1), anti-SDF-1 antibody (e.g., anti-hSDF-1/PBSF), rapamycin, cellulose, hydroxypropyl, and the like, Busulfan, cyclophosphamide, dacarbazine, hydroxyurea, mitotane, docetaxel, vinblastine, MGl 32, nimesulide, diclofenac, tenoxicam, indomethacin, acetylsalicylic acid, diflunisal (diflusinal), betamethasone, dexamethasone, deferoxamine mesylate, retinoic acid, heparin, pentoxifylline, streptokinase, TGF- β, TIMP-2, glucose, dextran T70, starch, quercetin dihydrate, caffeine, leflunomide, carrageenan (e.g., iota or lambda carrageenan), hydroxypropylcellulose, stachyose, and chondroitin sulfate A.
The therapeutic agent may also be an antineoplastic agent, an anti-inflammatory agent, an iron chelator, a triene macrolide antibiotic, a 3-hydroxy-3-methylglutaryl-coa reductase inhibitor, a retinoid, an antithrombotic agent, an anticoagulant, a plasminogen activator, a cytokine, a matrix metalloproteinase inhibitor, a tetracycline, an ACE inhibitor, a dextran, or a carrageenan, an alkylating agent, an antimetabolite, a ribonucleotide reductase inhibitor, a cytotoxic antibiotic, a taxane, a vinca alkaloid, or a protease inhibitor, a COX-2 inhibitor, a fenamate, an oxicam, an acetyl acid derivative, a salicylic acid derivative, or a corticosteroid.
As noted elsewhere, the various aspects, embodiments, features, etc. in this specification can be mixed and matched in any desired manner, as well as combined and substituted. Therefore, specific drugs and disease targets and the like in the context may be appropriately combined and the like, although they do not appear together in the same paragraph. Some discussion of certain suitable second drugs may be found in, for example, US patent No.6,812,220 issued on day 11/2 of 2004, US patent No.7,163,930 issued on day 16/1/2007, US published application No.20080063682, and PCT publication nos. wo2004105737 and wo2006032143, all of which are incorporated herein by reference.
In yet another aspect, the compositions and methods, etc., of the invention in this specification provide kits. The kit comprises a container containing the composition of the present specification and a label comprising instructions for the pharmaceutical use of the composition for inhibiting fibrous adhesions. The label may be a government approved label, for example an FDA approved label, such as the appropriate FDA standard on 7/1/2009. The container may be a vial configured to hold a drip agent or any other desired composition described herein. The label may additionally comprise instructions for pharmaceutical use of the composition for treating at least one of a non-fibrous adhesion disease or a non-fibrous adhesion condition.
The above and other aspects, features and embodiments are set forth in this application, including the following detailed description.
Detailed Description
General discussion of exemplary drugs
In certain embodiments, the compositions and methods, etc., of the invention in this specification include the use of selected modified fucoidans to treat, prevent, inhibit, etc., diseases or conditions such as fibrous adhesions (e.g., surgical adhesions), the selected modified fucoidans including specific combinations of components that generally include: a total carbohydrate content of more than about 40% w/w; a trehalose content of about 40-100% by weight of the total carbohydrate content; a galactose content of about 0-60% as a percentage of the total carbohydrate content; a sugar content excluding trehalose and galactose of about 0-20% as a percentage of total carbohydrate content; an acetyl group to trehalose monomer ratio of less than about 40%; a molecular weight distribution such that about 0 to 5,000g/mol of the modified fucoidan moiety comprises less than about 30% w/w; such that about 5,000-60,000 g/mol of the modified fucoidan moiety comprises a molecular weight distribution of less than about 50% w/w; such that about 60,000 to 200,000g/mol of the modified fucoidan fraction represents less than about 40% w/w of the molecular weight distribution; such that about 200,000 to 1,600,000g/mol of the modified fucoidan moiety comprises a molecular weight distribution of less than about 50% w/w; such that greater than about 1,600,000g/mol of the modified fucoidan moiety comprises a molecular weight distribution of less than about 50% w/w; a sulfate content of about 10-50% w/w; and/or a water content of less than about 20% w/w. The selected modified fucoidan also includes modified fucoidan that forms a solution having a pH of about 4 to about 8 when prepared as a 0.1% w/v solution. The compositions and methods, etc. may additionally comprise modified fucoidan for the treatment of: inflammatory diseases (including arthritis and peritonitis), ischemia, endotoxemia, keloid features/raised scars, keloids (keloid scars), dermatitis (including acanthotic dermatitis, seborrheic dermatitis (dandruff), dyshidrotic dermatitis (pompholyx), urticaria, vesicular dermatitis (bullous dermatitis), and papular urticaria), rosacea (e.g., dilated erythematous rosacea, pustular rosacea, excrescent erythematous rosacea, and ocular erythematous rosacea).
The composition may be a medical composition, which may be a pharmaceutical or nutritional composition. As used herein, pharmaceutical compositions refer to pharmacological agents (drugs) as well as medical devices that consist of or contain the compositions of the present specification. Pharmaceutical compositions are distinct from nutritional compositions, and medical compositions are considered products such as medical foods.
The composition may also include a second agent, for example, for treating, inhibiting, or preventing the formation of fibrous adhesions or other diseases discussed in this specification. Such adhesions may form following surgery, post trauma, or following radiation or chemotherapy, or as a result of any other reason, and may be applied to the tissues of an animal (including a human, dog, cat, horse, cow, or other mammal, or bird, reptile, or other animal) at a site suspected of having a fibrous adhesion, such as a site actually suffering from a fibrous adhesion, a site where a fibrous adhesion has excessively developed (e.g., due to exposure to radiation, surgery, disease, or injury), and at a site where a fibrous adhesion has developed or expanded. Unless otherwise expressly stated, each second listed agent non-exclusively includes that agent and all derivatives, salts and analogs thereof. The second agent may be administered in a different formulation to inhibit fibrous adhesions. These compositions may release an effective amount of the second drug only at the site of disease, if desired, in order to reduce toxicity that may be associated with systemic delivery of some of the compounds. These compositions may contain a polymeric formulation of the second agent (including all derivatives, salts and analogues thereof) as described herein, or other formulations if desired, which may provide slow release of the second agent at the site of a possible fibrous adhesion. The composition may be administered directly, systemically, or otherwise to the site as desired. In certain embodiments, the compositions of the present specification do not comprise antisense oligonucleotides or other oligonucleotide drugs (e.g., gene therapy nucleotides).
Embodiments of the present description may include identifying a fibrous or non-fibrous adhesion disease or condition, and then selecting and administering a composition comprising a composition of the present description. In some embodiments, the compositions and methods may further comprise selecting two or more of the agents of the present specification such that one has a primary effect against the disease or condition and the other has a primary effect. Exemplary non-fibrous adhesion diseases or conditions include cancer, PID, radiation exposure, mechanical and other injuries, arthritis, surgery, topical conditions, diseases and conditions of the gastrointestinal tract (e.g., those with a significant risk of obstruction or other mechanical rupture symptoms, etc.).
In certain embodiments, the anti-fibrous adhesion agents can be formulated with other compounds or compositions, for example, as ointments, solutions, creams, powder lotions (powder deposition), gels, sprays, mousses (mousses), coatings (coating), wraps (wrap), pastes, barriers (barriers), implants, microspheres, microparticles, membranes, granules, liquids, implant films, instillations, and the like. Routes and sites of administration include oral, systemic, intraocular, subcutaneous, intraperitoneal, intramuscular, intraarticular, intralesional, intravaginal, rectal, or topical (e.g., in the form of a patch).
The compositions of the present specification may be provided in a suitable container or vessel which in turn may be provided in a kit and may also have a label, preferably a label approved by an appropriate governmental regulatory agency (e.g., the U.S. food and drug administration). The label may contain instructions for the pharmaceutical use of the composition. The container may be, for example, a vial, and may be configured to provide a film, gel, drip, or other form of the composition discussed in this specification or otherwise desired.
The compound or composition administered with the anti-fibrous adhesion agent, which may be polymeric or non-polymeric, may act as a carrier and/or physical barrier. The compositions discussed in this specification are also aloneOr the drug (or any combination of drugs from the list of drugs discussed in this specification including fucoidan, modified fucoidan, or other modified fucoidan) in an aqueous solution, or a non-aqueous solution, or dispersed in suspension in a medium or carrier. Representative examples of polymeric carriers, barriers, and excipients include chitosan, polytetrafluoroethylene, poly (lactic acid), poly (ethylene vinyl acetate), poly (glycolic acid), copolymers of ethylene and vinyl acetate, methoxypolyethylene glycol, polycaprolactone, copolymers of lactic acid and glycolic acid, copolymers of poly (lactic acid) and poly (caprolactone), gelatin, collagen, cellulose, albumin, pluronic, poly (valerolactone), poly (anhydride), polysaccharides, alginic acid (e.g., alginate), hyaluronic acid, injectable excipients, other polymeric vehicles and copolymers, derivative mixtures and blends thereof. Representative examples of other suitable carriers include ethanol, glycols (including ethylene glycol, propylene glycol, or mixtures thereof)) A mixture of ethanol and glycol, isopropyl myristate or isopropyl palmitate, a mixture of ethanol and isopropyl myristate or isopropyl palmitate. The polymers themselves may provide antiblocking activity in certain compositions.
Exemplary second agents include NSAIDs, COX-2 inhibitors, nimesulide, fenamate (including meclofenamic acid, meclofenamate sodium, and diclofenac), oxicams (including tenoxicam), acetyl acid derivatives (including indomethacin), salicylic acid derivatives (including acetylsalicylic acid and diflunisal), pyrazolones (including phenylbutazone), corticosteroids (including dexamethasone), alkylating agents (including busulfan, cyclophosphamide, estramustine, cisplatin, and dacarbazine), antimetabolites (including methotrexate), ribonucleotide reductase inhibitors (including hydroxyurea), cytotoxic antibiotics (including mitotane), taxanes, topoisomerase inhibitors (including docetaxel), vinca alkaloids and analogs (including vinblastine), proteasome inhibitors (including MGl 32), iron chelators (including deferoxamine mesylate), 3-hydroxy-3-methylglutaryl-coenzyme a reductase inhibitors (including statins and simvastatin), retinoids and retinoid analogues (including all-trans retinoic acid), antithrombotic agents (heparin sodium and low molecular heparin), anticoagulants (including pentoxifylline), plasminogen activator (including streptokinase), cytokines (including transforming growth factor-beta (TGF-beta)), matrix metalloproteinase inhibitors, tissue inhibitors of matrix metalloproteinases (TIMP) (including TIMP-2), tetracyclines (including minocycline and doxycycline), Angiotensin Converting Enzyme (ACE) inhibitors (including captopril and enalapril, including salts thereof, such as enalapril maleate). Some other desirable drugs include: leflunomide (Arava), erythromycin, dextran sulfate alginic acid, glucose, dextran T70, starch, quercetin dihydrate, caffeine, iota carrageenan, lambda carrageenan, hydroxypropyl cellulose, stachyose and chondroitin sulfate A. A discussion of the second agent described above can be found, for example, in PCT publication No. wo 2006032143.
Fucoidan
Fucoidan (including fucoidan and modified fucoidan) is a high molecular weight sulfated polysaccharide extracted from brown algae, Percival, e.g., and McDowell, r.h., Chemistry and Enzymology of MarineAlgal Polysaccharides, pp.157-175(Academic Press, New York,1967), and can be found as is well known from other sources, such as the families of the order fucoidales and the family Laminariaceae (Laminariaceae), or from other seaweeds and seaweeds as well as echinoderms, sea cucumbers, sea urchins, or other sources as desired, including synthetic sources. Fucoidan (or fucoidin) refers to a modified fucoidan derived from seaweed or other sources. See USPA 2003064958. Fucoidan can be present alone or in mixtures, for example, with sugars such as xylose, galactose, glucose, and/or mannose. The sugars are known to be contained in seaweed and can be extracted together with modified fucoidan. Duarte, Maria ER., Cardoso, Marc A., Noseda, Miguel D., Cerezo, Alberto S., "Structural students on semiconductors from the brown felt Sargassum stenophyllum". CarbohydratedResearch: 2001(333): 281-29. Other sulfated modified fucoidans, including linear, branched, and linear sulfated modified fucoidans, are reported to have different anticoagulant activities (Pereira, M.S., J biol. chem.12:7656-67 (1999)).
Fucoidan (e.g., fucoidan) can be obtained from a variety of brown algae species, including but not limited to: brown algae (Adenocystis utraria), Ascophyllum nodosum (Ascophyllum nodosum), Chorda funiculosum (Chorda filum), Cladosporium rougherii (Cladosiphon okamuranus), Alternaria crassipes (Cystoseirabilis marina), Laminaria japonica (Ecklonia kurome), Fucus vesiculosus (Fucus evanescens), Fucus vesiculosus (Fucus vesiculosus), Hizikia fusiforme (Hizikia fusiforme), Kjellmaniella crassifolia (Kjellmaniella crassifolia), Laminaria brasiliensis (Laminarian brasiliensis), Laminaria crispa (Laminaria citrifolia), Laminaria japonica (Laminaria japonica), Laminaria japonica (Laminaria digitata), Undaripinula pinnata (Laminaria digitata), and Undaria pinnatifida (Undaripinia pinnatifida). The above species are all from the taxonomic class phaeophyceae, and most of them belong to the order Fucales and Laminariaceae.
Film
The medicaments discussed in this specification can be formulated into films suitable for direct administration to the tissues of animals, including humans, to treat fibrous adhesions. Desirable properties of the film include thinness, flexibility, ability to be handled, and ability to be placed against tissue. The individual drugs discussed in this specification can also be added to the polymer to form a film. The performance of the polymer film formulation can be enhanced by the addition of suitable excipients. In one embodiment, the drug may be combined with a hyaluronic acid polymer to make a membrane. Excipients that may be added include l-ethyl-3- [3- (dimethylamino) propyl ] carbodiimide (EDAC) and glycerol.
One embodiment in this specification is the addition of the drug to create a drug-loaded (therapeutic agent) film comprising 0.001% -99% w/w drug, 50% -99% w/w drug, 0.001% -50% w/w drug, 10% -50% w/w drug, 30% -40% w/w drug, 0.001% -10% w/w drug, 1% -10% w/w drug, 0.001% -1% w/w drug, 1% -5% w/w drug, 1% -2% w/w drug, or other concentrations discussed in this specification. One embodiment involves adding the drug and hyaluronic acid together, resulting in a 5% w/w drug-loaded membrane, the remainder of the membrane consisting of hyaluronic acid, glycerol and EDAC in a ratio of about 45:19: 3.
Gel
The various drugs discussed in this specification may be added to a viscous solution, which is referred to herein as a gel. Such gels can be administered to a body cavity of an animal, including a human, and are effective in inhibiting or preventing the formation of fibrous adhesions.
Desirable properties for a gel include being viscous enough to be applied to a particular site and remain attached thereto, so the gel should not flow under its own weight; and the gel may be applied to the preferred site by using a syringe or by injecting the gel through a needle. In one embodiment, the gel comprises a 5.5% w/v hyaluronic acid solution, wherein addition of the drug results in 0.001% -1% w/v gel, 1% -10% w/v gel, or 10% -50% w/v gel, or other concentrations as desired.
Drip agent
The various compositions discussed in this specification can also be dissolved or suspended in a liquid that can be administered to a body cavity of an animal (including a human) and used to inhibit, treat or prevent the formation of, including the acceleration of growth, of, etc., fibrous adhesions. Such formulations are referred to herein as instillation formulations. These formulations may be administered to a patient intraperitoneally, e.g., after a surgical procedure and to prevent post-surgical adhesion formation, or into/on any desired site of a wound or disease, etc. The liquid may be a solvent and may subsequently form a solution of the drug. In addition, the solvent for dissolving the drug may be water-based, and may be an electrolyte.
In some embodiments, the instillate solution is a substantially non-viscous liquid that is capable of reaching substantially all areas of a particular body lumen into which it is introduced, e.g., based on a viscosity substantially similar to water or substantially similar to a physiological saline solution. Desired mixtures may incorporate at least one drug discussed in this specification in the liquid to produce a solution at a concentration of about 0.0001% w/v to 1% w/v, 1% w/v to 2% w/v, 2% w/v to 5% w/v, 5% to 10% w/v, 10% w/v to 25% w/v, and 25% w/v to 50% w/v, or other concentration as desired. In some embodiments, the instillate solution can be first prepared as a concentrated solution (or suspension or sol, etc.) and then diluted (e.g., with a commercially available saline solution or ringer's lactate solution USP or other solution) to form a substantially non-viscous solution. The concentrated solution (or suspension or sol, etc.) may be prepared at a concentration of about 1% w/v to 25%, 1% w/v to 20% w/v, 1% w/v to 15% w/v, 1% w/v to 10% w/v, 5% w/v to 10% w/v, and 5% w/v, or other concentrations as desired.
Discussion of quantitative efficacy of anti-fibrous adhesion Agents
In one embodiment, the efficacy of a particular drug or combination of drugs can be evaluated based on the reduction in the average total adhesion value (strength x area; "TAV") of that drug or composition relative to a particular standard, such as a drug-loaded sodium hyaluronate membrane relative to a control or sodium hyaluronate membrane alone in a rat caecal lateral wall model for surgical fibrous adhesions. Other standards may include other membranes or solutions, etc., as well as other models such as a rabbit uterine horn model or efficacy in humans. In various embodiments, using the rat caecum lateral wall model for surgical fibrous adhesions, the average TAV of the drug may be less than or equal to 0.01%, 1%, 5%, 10%, 25%, 50%, or 75% of a control value (e.g., hyaluronate film alone). Among other measured parameters, the medicament may inhibit substantially all fibrous adhesion formation in a patient. Other exemplary embodiments
In some embodiments, the pharmaceutical composition may be a solution, gel, sol, or suspension and may have a total w/v fucoidan concentration of the composition of 0 to 10% w/v, 5% w/v, 0.001 to 1% w/v, 0.05% w/v, or 0.03% w/v.
In certain embodiments, the compositions or methods, etc., of the present invention include in the present specification treatment or inhibition, etc., with modified fucoidans (e.g., modified fucoidans including tailored fucoidans (modified fucoidans)) to provide enhanced efficacy and/or reduced toxicity and/or improved handling characteristics during processing, manufacture, transport, and/or administration to patients, including humans, animals, reptiles, and birds. Modified fucoidan includes modified fucoidan having white to off-white, white to light yellow, white to light orange, white to light green, and white to light brown appearances. The modified fucoidan further comprises a modified fucoidan having a total carbohydrate content of about 30-100% w/w, 40-90% w/w, 50-80% w/w, 37-75% w/w, and about 55-75% w/w. The modified fucoidan further comprises a trehalose in an amount of about 20-100%, about 30-100%, 31-71% w/w, about 40-100%, about 50-100%, about 60-100%, about 70-100%, about 80-100%, about 90-100%, about 40-80%, about 50-70%, and about 51-71% by weight of the total carbohydrate content.
The modified fucoidan further comprises a modified fucoidan having a galactose content of about 0-70%, 9-46% w/w, about 10-60%, about 20-50%, about 25-45%, and about 26-46% by weight of total carbohydrate content. The modified fucoidan further comprises a modified fucoidan having a sugar content excluding trehalose and galactose in a percentage of the total carbohydrate content of about 0 to 59% w/w, 0 to 40%, 0 to 30%, 0 to 20%, 0 to 15%, about 0 to 10%, about 0 to 6%, and about 0 to 5%. The modified fucoidan further comprises a modified fucoidan having a saccharide content of about 0-40%, 0-30%, about 0-20%, 0-15%, about 0-10%, about 0-6%, and about 0-5% excluding trehalose, in percentage of the total carbohydrate content.
The modified fucoidan further comprises a modified fucoidan having an acetyl group content of about 0-100% w/w, 0-70% w/w, 0-40% w/w, 0-36% w/w, 0-30% w/w, about 0-20% w/w, about 0-10% w/w, about 0-5% w/w, and about 0-2% w/w. The modified fucoidan further comprises a modified fucoidan having a molecular weight distribution such that the fraction of about 0 to 5,000g/mol is about 0 to 50% w/w, about 0 to 40% w/w, about 0 to 30% w/w, about 0 to 25% w/w, and about 0 to 20% w/w.
The modified fucoidan further comprises a modified fucoidan having a molecular weight distribution such that the fraction of about 0 to 5,000g/mol is less than about 0 to 25% w/w or 0 to 30% w/w. The modified fucoidan further comprises a modified fucoidan having a molecular weight distribution such that the fraction of about 5,000-60,000 g/mol is about 0-55% w/w, about 5-38% w/w, about 10-45% w/w, about 15-40% w/w, and about 17.5-37.5% w/w. The modified fucoidan further comprises a modified fucoidan having a molecular weight distribution such that the fraction of about 60,000-200,000 g/mol is about 0-60% w/w, about 0-50% w/w, about 0-40% w/w, about 5-35% w/w, and about 10-30% w/w. The modified fucoidan further comprises a modified fucoidan having a molecular weight distribution such that the fraction of about 200,000 to 1,600,000g/mol is about 0 to 60% w/w, about 0 to 50% w/w, 8 to 43% w/w, and about 10 to 40% w/w. The modified fucoidan further comprises a modified fucoidan having a molecular weight distribution such that the fraction of about 1,600,000g/mol and greater than 1,600,000g/mol is about 0-60% w/w, about 0-50% w/w, 1-33% w/w, and about 2.5-42.5% w/w. The modified fucoidan further comprises a sulfated content of about 0-60% w/w, about 10-50% w/w, and about 20-40% w/w. The modified fucoidan further comprises a water content of about 0-20% w/w, about 0-15% w/w, 14-40% w/w, or about 0-10% w/w. The modified fucoidan further comprises a modified fucoidan having a protein content of about 0-12% w/w, 0-10% w/w, about 0-5% w/w, and about 0-2% w/w. The modified fucoidan further comprises a modified fucoidan that when formulated in a 0.1% w/v solution forms a solution having a pH of about 4 to 8, about 5.5 to 8, about 6 to 8, and about 6 to 7.5.
Examples
Example 1: the selected fucoidan is in solution formulation for safety and efficacy in preventing surgical adhesions in horses undergoing laparotomy and jejunostomy.
Fucoidan
Fucoidan is extracted from brown algae Undaria pinnatifida (from Australia).
Methods and materials for fucoidan analysis
Visual inspection was used to determine the appearance of fucoidan. Specific optical rotation was determined by optical rotation using 3% fucoidan solution according to USP CSA method <781 >. The residues of incandescence (sulfation) were determined according to the USP CSA method. Kinematic viscosity was measured using an Ubbelodhee viscometer. The individual sugar monomer content and total carbohydrate content were determined by derivatization and analysis with gas chromatography coupled with a mass selective detector using electron impact ionization mode. The molecular weight distribution was determined by gel permeation chromatography. And measuring the content of the sulfuric ester of the fucoidan by using an inductively coupled plasma spectrometry. The loss on drying was determined at about 105 degrees celsius. The pH of the 0.1% fucoidan solution was measured according to USP CSA method.
Methods and materials for fucoidan efficacy and safety during equine abdominal surgery
Twelve horses donated for reasons unrelated to gastrointestinal tract or abdomen were used. These horses were randomly grouped and assigned to 1 of 2 experimental groups: (1) fucoidan solution and (2) control LRS (6 pimari per group). Fucoidan solutions and control LRS were prepared by mixing 50mL fucoidan concentrate containing 2.5g fucoidan or 50mL LRS, respectively, into 5L LRS bags and heated to about body temperature prior to surgery.
Horses were treated perioperatively with flunixin meglumine (flunixin meglumine) and an antimicrobial agent. A simple abdominal exploration was performed by a 20-cm ventral midline abdominal wall incision. At the jejunal segment 10 and 5-arcuate orifice towards the ileum, a 1-cm full thickness circumferential wedge was cut and anastomosed using 2-0poly (lactic-co-glycolic acid) 910(2-0polyglactin 910) in a 2-layer simple continuous mode. The oral anastomosis site was used for mechanical testing and the reverse oral site for histological evaluation. Prior to closure of the white line, 5L fucoidan solution or control LRS was injected into the abdomen. White lines were juxtaposed in a simple continuous mode using #2 pla-glycolic acid 910. At the front of the white line closure, the continuous pattern was interrupted at 8cm and the segment was used for mechanical testing. The 12cm tail was used for histological evaluation. Subcutaneous tissue was apposed in a simple continuous mode using 2-0 pla-glycolic acid 910.
After surgery, the feed was gradually reintroduced over 48 hours. Physical examinations were performed every 12 hours. Horses were monitored for signs of colic and checked for postoperative reflux (defined as reflux of more than 1L per 8 hours). Incisions were subjectively graded for edema, palpation pain, pus discharge, and dehiscence. Examination of complete blood cell counts, plasma chemistry and coagulation characteristics was performed on day 1, 2, 6 and 10 before and after surgery. Horses were euthanized on day 10 post-surgery. Necropsy and pathology results are included in the accompanying excerpts.
Horses were euthanized on day 10 post-surgery. Necropsy was performed to assess healing and signs of adhesion or infection. The anterior half of the white line and the anastomotic site were wrapped in sterile saline and refrigerated for immediate mechanical testing. The latter half of the white line and the retrostomy were fixed in 10% neutral buffered formalin for histological evaluation.
The white line tension was measured, the load at break (newtons, N) was recorded, and then the white line length (N/cm) was corrected. The anastomotic burst pressure (mmHg) was recorded and burst wall tension (dynes/cm) was calculated. The anastomoses and incisions were stained with hematoxylin and eosin and the inflammation was graded. Continuous data was analyzed using one-way analysis of variance. Significance level was taken as p < 0.05.
Results of fucoidan analysis
Fucoidan was determined to have the following properties: white to off-white powder in appearance; the specific optical rotation is minus 68.4 degrees; blazing residueSlag about 25.9%; kinematic viscosity of about 2.05mmΛ2/s; a carbohydrate content of about 60.5%; trehalose content is about 52% as a percentage of the total carbohydrate content; a galactose content of about 48% as a percentage of the total carbohydrate content; the remaining sugar monomer content (sum of all sugar monomer content minus trehalose and galactose content) is about less than 1% as a percentage of the total carbohydrate content; a molecular weight distribution of about 0 to 5,000g/mol of about 8.4%, a molecular weight distribution of about 5,000 to 60,000g/mol of about 13.4%, a molecular weight distribution of 60,000 to 200,000g/mol of about 26.5%, a molecular weight distribution of 200,000 to 1,600,000g/mol of about 38.7%, and a molecular weight distribution of greater than about 1,600,000g/mol of about 13.2%; the sulfate content was about 30.6%; a loss on drying of about 3.7%; the pH of the 0.1% solution was about 6.9.
Results of fucoidan efficacy and safety during equine abdominal surgery
No differences were observed between experimental groups for heart rate or rectal temperature during the pre-operative or 10 day study period. No difference in the number of colic events was observed between the experimental groups. 2 horses in the fucoidan solution group and 3 horses in the LRS control group had post-operative reflux (post-operative reflux). The volume of postoperative reflux (23L in one animal from day 1 to day 4; and 139L in another animal from day 2 to day 5) was greater in the horses in the fucoidan group than in the LRS control group (9.5L in one animal within the first 24 hours of surgery; 4L in the second animal once on day 3 after surgery; and 5L in the third animal within the first 72 hours of surgery) and all horses were returned to normal. No signs at necropsy indicated the condition caused by postoperative reflux. Reflux and feces were negative for Salmonella spp. One horse in the control group developed a notch infection, while none of the horses in the fucoidan group had a notch infection (no statistical difference between the treatment groups). There were no signs of peritonitis in any of the horses. No differences were observed between experimental groups for fibrinogen concentration, platelet count, activated partial thromboplastin time, gamma-glutamyltransferase, aspartate aminotransferase or creatinine concentration. The difference in leukocyte and neutrophil counts, antithrombin III, prothrombin time and hematocrit was determined between experimental groups at several time points; however, the values are generally within the normal range. Although fucoidan is a sulfated polysaccharide (like heparin), treatment with fucoidan does not negatively affect any coagulation parameters.
One horse in the control LRS group had adhesions between the spleen and the body wall at the site of the incision infection. There were no additional adhesions (fucoidan group did not have any adhesions). No differences in anastomosis and incision healing between groups were found based on gross necropsy. Horses in the fucoidan solution group had significantly greater anastomotic burst pressure (262 ± 52 versus 206 ± 12mmHg, p ═ 0.03) than horses in the LRS control group, indicating that fucoidan might improve healing of the anastomotic site. Horses in the fucoidan solution group had a higher tendency to have higher intestinal burst wall tension (1,104,000 + -270,000 vs 941,000 + -189,000 dynes/cm) than horses in the LRS control group; however, this did not reach statistical significance (p ═ 0.29). There was no difference in tensile load at the time of white line break between the groups (67 ± 15 and 64 ± 21N/cm for fucoidan solution and LRS, respectively, with p ═ 0.81). Based on our analysis, there was no histological difference in the degree of inflammation at the anastomoses or white line incisions.
Fucoidan concentrates and fucoidan solutions are simple and easy to use in a clinical setting. Fucoidan reduced the number of animals with adhesions from about 17% in the control LRS to no animals with adhesions in the fucoidan group. Mechanical testing showed that the fucoidan solution did not impair anastomosis or incision healing, if anything, i.e., the strength of the anastomosis and incision on day 10 was greater than that of the control horses. Fucoidan solution is safely administered intraperitoneally during the laparotomy and anastomosis of horses; and we speculate that fucoidan solutions may have improved anastomotic healing.
Example 2: safety and efficacy of fucoidan solution formulations for preventing surgical adhesions in dwarf horses undergoing intestinal abrasion (intestinal abrasion)
Fucoidan
Fucoidan is extracted from brown algae Undaria pinnatifida (from Australia).
Methods and materials for fucoidan analysis
Visual inspection was used to determine the appearance of fucoidan. The individual sugar monomer content and total carbohydrate content were determined by derivatization and analysis with gas chromatography coupled with a mass selective detector using electron impact ionization mode. The molecular weight distribution was determined by gel permeation chromatography. And measuring the content of the sulfuric ester of the fucoidan by using an inductively coupled plasma spectrometry. Acetyl content was determined by IH NMR (providing a quantitative determination of acetyl, integration of peaks at 1.6 and 2.5ppm, and ratio is recorded as degree of acetylation).
Methods and materials for fucoidan efficacy and safety during pongam abdominal surgery
Neonatal ponies were surgically operated to induce adhesions using serosal abrasion plus gut sutures at four locations on the jejunum. Before closing the surgical site, 600mL of treatment solution was administered intraperitoneally and the surgical site and incision were sutured. The treatment group consisted of control ringer's injection usp (LRS) (n ═ 6) and 0.03% fucoidan (n ═ 6) dissolved in LRS. A laparoscopic secondary examination was performed after 10 days and the number of adhesions per pony and the characteristics of each adhesion (simple vs. complex) were assessed. Adhesions were scored for severity and the total severity score for each treatment group was determined. By comparing animal body weight and hematological parameters (including white blood cell differentiated complete blood count, coagulation and fibrinogen) between the two treatment groups; and study of the toxicity of the treatments by observing signs of toxicity (posture, appetite, rectal temperature and heart rate, etc.).
Results of fucoidan analysis
The fucoidan total carbohydrate content was determined to be 51.1 + -5.6% w/w of fucoidan. The respective sugar content of fucoidan was determined as shown in the following table:
| sugar monomers | Average amount (percentage of total carbohydrate) | Standard deviation (percentage of total carbohydrate) |
| Trehalose | 48.9 | 2.2 |
| Galactose | 47.1 | 2.3 |
| Glucose | 1.4 | 0.4 |
| Mannose | 1.1 | 0.3 |
| Xylose | 0.8 | 0.2 |
| Rhamnose | 0.6 | 0.1 |
The molecular weight distribution of fucoidan was determined as shown in the following table:
molecular weight
The fucoidan sulfate content is about 32.9 + -0.9% w/w. The fucoidan acetyl content was determined such that the ratio of acetyl groups to trehalose was determined to be 0 to 0.3%.
Results of fucoidan efficacy and safety during pongam abdominal surgery
The fucoidan solution is simple and easy to use in a clinical setting. Ponies that had been surgically treated and treated with 600ml of 0.03% w/v fucoidan solution had mean numbers of adhesions and total severity scores of 2.2 ± 1.2 and 3.5 ± 1.5, respectively, which were all significantly reduced compared to the control LRS (6.2 ± 2.5 and 14.5 ± 3.8, respectively). The 13 adhesions observed in the fucoidan-treated group were not complex, while 20 of the 37 adhesions observed in the control-treated group were complex. No death or painless death at moribund was observed. No signs of toxicity were observed in any animals. There were no differences in hematological parameters between treatment groups, except for several small statistically significant differences in the normal range that were considered clinically irrelevant. One of the ponies treated with fucoidan solution had a higher than normal range of lobular nuclear neutrophil counts at days 4, 6,8 and 10. Although the 0-day lobulated nuclear neutrophil value was not obtained for this animal, the banded nuclear neutrophil count at this initial time point was higher than normal and therefore not considered a result of the treatment. The fucoidan solution is easy to apply and safely prevents the formation of adhesions in a pongamic abdominal serosal abrasion model of surgical adhesions.
Example 3: safety and efficacy of fucoidan solution formulations for preventing surgical adhesions in rabbits undergoing uterine horn surgery
Fucoidan
Several fucoidan extracts were prepared from brown algae Undaria pinnatifida (from Australia).
Methods and materials for fucoidan analysis
Visual inspection was used to determine the appearance of fucoidan. The molecular weight distribution was determined by gel permeation chromatography. And measuring the content of the sulfuric ester of the fucoidan by using an inductively coupled plasma spectrometry. Acetyl content was determined by IH NMR (providing a quantitative determination of acetyl, integration of peaks at 1.6 and 2.5ppm, and ratio is recorded as degree of acetylation).
Methods and materials for fucoidan efficacy and safety during abdominal surgery in rabbits
Rabbits were intramuscularly anesthetized with 55mg/kg ketamine hydrochloride and 5mg/kb xylazine (Rompum). After preparation for sterile surgery, a midline laparotomy was performed. The uterine horn was removed from the abdomen and injured by abrading the serosal surface until punctate bleeding had formed. Ischemia of both uterine horns was induced by removal of collateral blood supply. The remaining blood supply to the uterine horn is the ascending branch of the uterine vaginal artery supply to the myometrium. The uterine horn was then returned to its normal anatomical position and the midline sutured with 3-0 Vicryl. When the incision is closed, a purse string suture is made around the catheter placed in the incision. Through the catheter, either no substance (surgical control) (n ═ 5) was administered, or 45mL of lactated ringer's injection USP (control LRS) (n ═ 5) or 0.03% w/v fucoidan (fucoidan solution) (n ═ 5) in lactated ringer's injection USP was placed in the abdomen. The last suture is then closed.
After 7 days, the rabbits were sacrificed and the percentage of area of uterine horn adhering to each organ was determined. In addition, the strength of adhesion was scored using the following system:
0 ═ no adhesion
1-mild, easily dissected adhesions
2-moderate adhesion; non-dissectable, non-lacerated organs
3, dense adhesion; non-dissectable, tearing organs when removed
In addition, each rabbit was given a total score taking into account all the information above. The following scoring system was used:
rabbits were scored by two independent observers blinded to previous animal treatment. A higher score is given if there is a discrepancy in the scores for the individual animals.
The total score was analyzed by rank order analysis and rank variance (variance on the ranks) analysis. The percentage of area of uterine horn that was adherent to multiple organs was compared by Student's t test or one-way analysis of variance.
Results of fucoidan analysis
The results of the fucoidan appearance, sulfate content and acetyl content analyses are included in the table below.
The results of the molecular weight distribution analysis of fucoidan are included in the following table.
Fucoidan efficacy, safety and ease of use results during abdominal surgery in rabbits
The fucoidan solution is simple and easy to use during surgery. The following table summarizes the efficacy of each fucoidan extract relative to surgical controls and control LRS solutions.
| Treatment of | Total adhesion score | Non-adhesive site (%) |
| Surgical contrast | 3.1±0.4 | 5.0 |
| Control LRS | 2.8±0.3 | 10.0 |
| Fucoidan extract # AA | 2.2±0.6 | 20.0 |
| Fucoidan (I) and its preparation methodSugar extract # B | 1.5±0.4 | 32.5 |
| Fucoidan extract # C | 1.4±0.8 | 40.0 |
| Fucoidan extract # D | 1.6±0.4 | 25.0 |
| Fucoidan extract # E | 1.2±0.3 | 42.5 |
| Fucoidan extract # F | 2.0±0.6 | 17.5 |
| Fucoidan extract # G | 1.3±1.4 | 37.5 |
| Fucoidan extract # H | 1.8±0.4 | 22.5 |
| Fucoidan extract # I | 1.7±0.6 | 15.0 |
No signs of toxicity were observed in any of the fucoidan solution-treated animals.
Example 4: a plurality of fucoidans: analyzing; safety in rabbits; efficacy and safety in rabbits undergoing surgery; efficacy and safety in rabbits undergoing surgery relative to sodium carboxymethylcellulose
Fucoidan
Fucoidan is extracted from Laminaria japonica (Laminaria japonica) of brown algae (from China), Laminaria japonica (from Europe), and Undaria pinnatifida (from Korea). As described in detail below, all 3 fucoidans were analyzed; all 3 fucoidans were prepared as solutions and tested for safety in rabbits at 16.7 and 50mg/kg body weight; all 3 fucoidans were prepared as solutions and tested for efficacy and safety in rabbits at 5mg/kg body weight; and a fucoidan (Laminaria japonica) prepared as a solution (5mg/kg body weight) and comparing the efficacy and safety in rabbits with sodium carboxymethylcellulose prepared as a gel.
Fucoidan analysis-methods and materials
Visual inspection was used to determine the appearance of fucoidan. And measuring the content of the sulfate by using inductively coupled plasma. The loss on drying was determined at about 105 degrees celsius. The pH of the 0.1% fucoidan solution was measured according to USP CSA method. Protein content was determined using ultraviolet-visible (UV-VIS) detection. The content of trehalose, galactose and other sugar monomers and the content of total carbohydrate are determined by ultra performance liquid chromatography with UV-VIS detection. The molecular weight distribution was determined by refractive index detection using gel permeation chromatography.
Fucoidan analysis-results
The results of the fucoidan appearance, sulfate content, and protein content analyses are included in the table below.
The results of the analysis of the total carbohydrate, trehalose monomer and galactose monomer content of fucoidan are included in the table below.
The results of the molecular weight distribution analysis of fucoidan are included in the following table.
Fucoidan safety-methods and materials in rabbits
Toxicity of each fucoidan in lactated forest format injection usp (lrs) was characterized after an observation period of 14 days following a single intraperitoneal injection into new zealand white rabbits.
The group assignments for the rabbits used in the study are summarized in the following table.
Note that 2 male rabbits and 2 female rabbits were initially assigned to group 5. However, for logistical reasons (logical reasons), 2 male rabbits were reassigned to groups 1 and 3, and 1 female rabbit was reassigned to group 2.
During the study, animals were observed for possible death and clinical signs. Body weight and food consumption were recorded. Blood samples were collected for clinical pathology evaluation (hematology, clinical chemistry and coagulation). Blood samples collected for toxicokinetics were stored for possible analysis. Necropsy was performed after the observation period and the overall observations were recorded.
Fucoidan safety-results in rabbits
There were no deaths in this study. Adverse clinical signs considered to be associated with the administration of 3 different fucoidan test items (16.7 or 50mg/kg) were not recorded during the study and there was no significant difference in weight gain between the treated groups and the controls. Occasional reductions in appetite were recorded in all animals before treatment began and therefore were dose independent.
After administration of 3 different fucoidan test items (16.7 or 50mg/kg), there was no effect associated with the test items with food consumption.
Hematological parameters indicative of the effect of the fucoidan test article were not considered to be changed and cytomorphological examination did not reveal any findings of toxicological significance. No change in coagulation parameters indicative of the effect of the test article was considered and no changes recorded in serum chemistry parameters indicated the treatment effect of 3 different fucoidan test articles at 16.7 or 50 mg/kg.
It is considered to indicate that there is no change in organ weight associated with the treatment.
There were no gross pathological findings in the treated animals that were considered indicative of the effect of treatment with either fucoidan test article.
In summary, treatment of rabbit groups with 3 different fucoidan test items extracted from kelp, kelp north and undaria pinnatifida at dose levels of 16.7 and 50mg/kg did not result in death or treatment-related changes, and there were no gross pathological findings in animals believed to be indicative of the effects of either test item.
Fucoidan efficacy and safety in rabbits undergoing abdominal surgery-methods and materials
New Zealand white rabbits were subjected to surgical procedures to induce adhesion formation between the uterine horn and the abdominal wall. Before closing the surgical site, 16.7mL/kg body weight of the treatment solution was administered intraperitoneally, and the surgical site and incision were sutured. The treatment group consisted of 0.03% w/v fucoidan (5mg fucoidan) extracted from kelp, kelp north and undaria pinnatifida per kg body weight of lactatein format injection usp (LRS) and LRS alone (control) (n ═ 4 per group). After 14 days, the animals were euthanized and any adhesions formed between the uterine horn and the lateral wall (referred to as the uterine adhesion score) or elsewhere in the abdomen (referred to as the abdominal adhesion value) were evaluated according to standard scoring methods. Toxicity of the treatments was studied by: comparing animal body weight and incision thickness between groups; comparing values of blood biochemical and coagulation parameters between pre-operative and post-operative groups at 24 hours and day 7; and observing clear signs of toxicity.
Fucoisan efficacy and safety-outcome in rabbits undergoing abdominal surgery
A reduction in uterine adhesion values (18.4 ± 7.8 vs. 3.1 ± 1.7, 1.3 ± 1.0 and 2.3 ± 0.5, respectively) was recorded in the groups of kelp, kelp northern and undaria pinnatifida fucoidan solutions relative to the control LRS. Significant reductions in abdominal adhesion values were recorded in the groups of kelp, kelp northern and undaria pinnatifida fucoidan solutions relative to the control LRS (2.5 ± 1.0 vs. 0.9 ± 0.3, 0.8 ± 0.3 and 0.3 ± 0.3, respectively). No death or painless death at moribund was observed. No differences in animal body weight and incision thickness were observed between the fucoidan solution and LRS, and no obvious signs of toxicity were observed in any of the animals. No difference in complete blood count, blood differentiation or coagulation parameters was observed between groups at any time point. Differences in alanine Aminotransferase (ALT) levels between control LRS and undaria pinnatifida were recorded (23.0. + -. 1.8 vs 16.8. + -. 1.0IU/L, respectively). The difference in ALT is likely to be clinically irrelevant. No other differences in any other blood chemistry parameters were observed between groups at any time point. In conclusion, a 0.03% w/v fucoidan solution prepared from kelp, kelp northern kelp and undaria pinnatifida, at 16.7mL per kg body weight, was effective in preventing adhesion formation in the rabbit uterine horn model of surgical adhesions and showed no signs of toxicity.
Efficacy and safety of fucoidan relative to sodium carboxymethylcellulose in rabbits undergoing abdominal surgery-methods and materials
Sodium carboxymethylcellulose (SCMC) solution is a viscous gel, and some veterinary surgeons either Intraperitoneally (IP) instill the SCMC solution immediately prior to closure of the abdominal wall during equine abdominal surgery, or use a portion of the SCMC solution during surgery to "enema" and IP instill the remainder of the SCMC solution immediately prior to closure in an attempt to reduce the formation of post-operative adhesions. The objective of this study was to compare the efficacy and safety of SCMC solution with fucoidan solution using an in vivo model of surgical adhesions after IP administration of the test article immediately prior to closure, or after "irrigating the uterine horn" with a portion of the test article and IP administration of the remainder of the test article immediately prior to closure during surgery, or after combined administration of both test articles. SCMC type 7H35F PH was obtained from Hercules Inc. New Zealand white rabbits were subjected to surgical procedures to induce adhesion formation between the uterine horn and the abdominal wall. Each animal received a total of 16.7mL of test article per kg of body weight (animals receiving the combination of test articles received a total of 16.7mL of each test article per kg of body weight). Three different test articles were used in this study: lactatein format injection usp (lrs) alone (control solution); 1% w/v SCMC in water for injection USP, equivalent to 167mg SCMC/kg body weight (SCMC solution); and 0.03% w/v fucoidan in LRS (from Laminaria japonica), corresponding to 5mg fucoidan/kg body weight (fucoidan solution). 6 treatment groups (6 for each group) were included in the study. Three groups consisting of LRS, fucoidan or SCMC were instilled intraperitoneally before closing the abdominal wall. In both groups, the fucoidan solution or SCMC was dispensed such that 5mL of the test article was applied to the uterine horn, while the remainder was instilled prior to closure. In one group, both SCMC and fucoidan were administered, with 5mL of SCMC solution administered to the uterine horn, and the remainder instilled with fucoidan prior to closure. After 14 days, the animals were euthanized and any adhesions formed between the uterine horn and the lateral wall (referred to as uterine adhesion values) or elsewhere in the abdomen (referred to as abdominal adhesion values) were evaluated according to standard scoring methods. Toxicity of the treatments was studied by: comparing animal body weight and incision thickness between groups; comparing values of blood biochemical and coagulation parameters between pre-operative and post-operative groups at 24 hours and day 7; and observing clinical signs of significant toxicity.
Efficacy and safety of fucoidan relative to sodium carboxymethylcellulose in rabbits undergoing abdominal surgery-results
No difference in uterine adhesion scores was observed between the control LRS and the two SCMC solution group treatments (20.0 ± 2.8, 19.3 ± 4.1, and 17.3 ± 5.0, respectively). The SCMC plus fucoidan solution combination treated group resulted in a significant reduction in uterine adhesion score relative to the control LRS and both SCMC solution treated groups (6.1 ± 5.2). Both fucoidan solution-treated groups resulted in a significant reduction in uterine adhesion scores (2.4 ± 1.1 and 3.0 ± 1.8) relative to all other treatment groups. No difference in abdominal adhesion scores was observed between the control LRS, the two SCMC solution treated groups, and the SCMC solution plus fucoidan solution combination treated group (2.6 ± 2.4, 2.4 ± 1.4, 2.3 ± 1.3, and 2.1 ± 1.1, respectively). Both fucoidan solution-treated groups resulted in a significant reduction (0.3 ± 0.5 and 0.2 ± 0.4) in abdominal adhesion score relative to all other treated groups. There was no difference in uterine or abdominal adhesions between the two fucoidan solution-treated groups. No differences in animal body weight or incision thickness were observed between the post-treatment group and the control LRS group, and no obvious signs of toxicity were observed in any of the animals. No difference in complete blood count, blood differentiation or blood chemistry parameters was observed between the control LRS and the treated groups at any time point.
An increase in Activated Partial Thromboplastin Time (APTT) was observed between pre-operative and 24 hour values in the control LRS group, the two SCMC solution treated groups, and the SCMC plus fucoidan solution combined treated group, but not in the two fucoidan groups. In summary, SCMC solution treatment had no effect on adhesions in rabbit uterine horn lateral wall surgical adhesion models; treatment with a combination of SCMC solution plus fucoidan (from laminaria japonica) solution resulted in a reduction of uterine horn adhesion value of about 70%, but no effect on abdominal adhesion value; and fucoidan (from laminaria japonica aresch) solution treatment resulted in a reduction of adhesion formation of about 90%. No difference in efficacy was observed between IP instillation of the fucoidan solution immediately prior to closure or use of a portion of the fucoidan solution to "irritate the uterine horn" and IP administration of the remaining portion immediately prior to closure. Fucoidan solutions alone or in combination with SCMC showed no signs of toxicity. Example 5: efficacy of selected fucoidan solution formulations on horses with peritonitis
Fucoidan was extracted from brown algae Undaria pinnatifida and was determined to have the properties given in the following table using the analytical techniques described in the above examples:
| testing | Results |
| Appearance of the product | White to off-white powder |
| Total carbohydrate content | 61％w/w |
| Trehalose content | 52% w/w of total carbohydrates |
| Galactose content | 48% w/w of total carbohydrates |
| Other sugar monomers | Less than 1% w/w of total carbohydrate |
| Molecular weight of carbohydrate | |
| <5k g/mol | 8％w/w |
| 5k-20k g/mol | 2％w/w |
| 20k-60k g/mol | 11％w/w |
| 60k-200k g/mol | 27％w/w |
| 200k-1,100k g/mol | 35％w/w |
| 1,000k-1,600k g/mol | 4％w/w |
| >1,600k g/mol | 13％w/w |
| Content of sulfuric acid ester | 31％w/w |
| Ratio of acetyl to trehalose (%) | Less than 2 percent |
| Water content | Less than 10% w/w |
| pH of 0.1% w/v solution | 7 |
Fucoidan solution was prepared by dissolving 5g fucoidan in 50mL lactated ringer's injection USP, sterilizing with autoclaving and allowing the solution to cool to ambient temperature. Horses suffering from peritonitis (including symptoms of peritonitis such as abdominal pain, abdominal tenderness, and abdominal strain) were injected intravenously with a fucoidan solution by intravenous drip for 30 minutes. The peritonitis and peritoneal inflammation of horses were reduced after treatment with the fucoidan solution.
Example 6: fucoisan solution formulation efficacy on horses with peritonitis
Fucoidan was extracted from brown algae Undaria pinnatifida and was determined to have the properties given in the following table using the analytical techniques described in the above examples:
| testing | Results |
| Appearance of the product | White to off-white powder |
| Total carbohydrate content | 61％w/w |
| Trehalose content | 52% w/w of total carbohydrates |
| Galactose content | 48% w/w of total carbohydrates |
| Other sugar monomers | Less than 1% w/w of total carbohydrate |
| Molecular weight of carbohydrate | |
| <5k g/mol | 8％w/w |
| 5k-20k g/mol | 2％w/w |
| 20k-60k g/mol | 11％w/w |
| 60k-200k g/mol | 27％w/w |
| 200k-1,100k g/mol | 35％w/w |
| 1,000k-1,600k g/mol | 4％w/w |
| >1,600k g/mol | 13％w/w |
| Content of sulfuric acid ester | 31％w/w |
| Ratio of acetyl to trehalose (%) | Less than 2 percent |
| Water content | Less than 10% w/w |
| pH of 0.1% w/v solution | 7 |
Fucoidan solution was prepared by dissolving 5g fucoidan in 50mL lactated ringer's injection USP, sterilizing with autoclaving and allowing the solution to cool to ambient temperature. Horses suffering from peritonitis (including symptoms of peritonitis, such as abdominal pain, abdominal tenderness, and abdominal strain) were administered 50mL of a fucoidan solution by intra-abdominal injection. The peritonitis and peritoneal inflammation of horses were reduced after treatment with the fucoidan solution.
Example 7: fucoigan solution preparation for horse with ischemia
Fucoidan was extracted from brown algae Undaria pinnatifida and was determined to have the properties given in the following table using the analytical techniques described in the above examples:
| testing | Results |
| Appearance of the product | White to off-white powder |
| Total carbohydrate content | 61％w/w |
| Trehalose content | 52% w/w of total carbohydrates |
| Galactose content | 48% w/w of total carbohydrates |
| Other sugar monomers | Less than 1% w/w of total carbohydrate |
| Molecular weight of carbohydrate | |
| <5k g/mol | 8％w/w |
| 5k-20k g/mol | 2％w/w |
| 20k-60k g/mol | 11％w/w |
| 60k-200k g/mol | 27％w/w |
| 200k-1,100k g/mol | 35％w/w |
| 1,000k-1,600k g/mol | 4％w/w |
| >1,600k g/mol | 13％w/w |
| Content of sulfuric acid ester | 31％w/w |
| Ratio of acetyl to trehalose (%) | Less than 2 percent |
| Water content | Less than 10% w/w |
| pH of 0.1% w/v solution | 7 |
Fucoidan solution was prepared by dissolving 5g fucoidan in 50mL lactated ringer's injection USP, sterilizing with autoclaving and allowing the solution to cool to ambient temperature. Horses with ischemia were injected intravenously with 50mL fucoidan solution by 30 min intravenous drip. Following treatment with fucoidan solution, the horses had reduced ischemia.
Example 8: efficacy of fucoidan solution formulations on horses with reperfusion injury
Fucoidan was extracted from brown algae Undaria pinnatifida and was determined to have the properties given in the following table using the analytical techniques described in the above examples:
| testing | Results |
| Appearance of the product | White to off-white powder |
| Total carbohydrate content | 61％w/w |
| Trehalose content | 52% w/w of total carbohydrates |
| Galactose content | 48% w/w of total carbohydrates |
| Other sugar monomers | Less than 1% w/w of total carbohydrate |
| Molecular weight of carbohydrate | |
| <5k g/mol | 8％w/w |
| 5k-20k g/mol | 2％w/w |
| 20k-60k g/mol | 11％w/w |
| 60k-200k g/mol | 27％w/w |
| 200k-1,100k g/mol | 35％w/w |
| 1,000k-1,600k g/mol | 4％w/w |
| >1,600k g/mol | 13％w/w |
| Content of sulfuric acid ester | 31％w/w |
| Ratio of acetyl to trehalose (%) | Less than 2 percent |
| Water content | Less than 10% w/w |
| pH of 0.1% w/v solution | 7 |
Fucoidan solution was prepared by dissolving 5g fucoidan in 50mL lactated ringer's injection USP, sterilizing with autoclaving and allowing the solution to cool to ambient temperature. Immediately before the reperfusion treatment, each of the horses with ischemia to be subjected to reperfusion treatment was intravenously injected with 50mL of fucoidan solution or control lactating forest format injection USP by 30 minutes intravenous drip. The reperfusion injury was reduced in horses receiving fucoidan solution compared to horses receiving control lactated ringer's injection USP.
Example 9: efficacy of fucoidan solution formulations on horses with endotoxemia
Fucoidan was extracted from brown algae Undaria pinnatifida and was determined to have the properties given in the following table using the analytical techniques described in the above examples:
| testing | Results |
| Appearance of the product | White to off-white powder |
| Total carbohydrate content | 61％w/w |
| Trehalose content | 52% w/w of total carbohydrates |
| Galactose content | 48% w/w of total carbohydrates |
| Other sugar monomers | Less than 1% w/w of total carbohydrate |
| Molecular weight of carbohydrate | |
| <5k g/mol | 8％w/w |
| 5k-20k g/mol | 2％w/w |
| 20k-60k g/mol | 11％w/w |
| 60k-200k g/mol | 27％w/w |
| 200k-1,100k g/mol | 35％w/w |
| 1,000k-1,600k g/mol | 4％w/w |
| >1,600k g/mol | 13％w/w |
| Content of sulfuric acid ester | 31％w/w |
| Ratio of acetyl to trehalose (%) | Less than 2 percent |
| Water content | Less than 10% w/w |
| pH of 0.1% w/v solution | 7 |
Fucoidan solution was prepared by dissolving 5g fucoidan in 50mL lactated ringer's injection USP, sterilizing with autoclaving and allowing the solution to cool to ambient temperature. Horses suffering from endotoxemia, including symptoms of endotoxemia such as septic shock, are injected intravenously with 50mL of fucoidan solution by 30 minutes of intravenous drip. The symptoms of endotoxemia and endotoxemia in horses were reduced after treatment with the fucoidan solution.
Example 10: efficacy of fucoidan solution formulations for humans with keloid characteristics
Fucoidan was extracted from brown algae Undaria pinnatifida and was determined to have the properties given in the following table using the analytical techniques described in the above examples:
| testing | Results |
| Appearance of the product | White to off-white powder |
| Total carbohydrate content | 61％w/w |
| Trehalose content | 52% w/w of total carbohydrates |
| Galactose content | 48% w/w of total carbohydrates |
| Other sugar monomers | Less than 1% w/w of total carbohydrate |
| Molecular weight of carbohydrate | |
| <5k g/mol | 8％w/w |
| 5k-20k g/mol | 2％w/w |
| 20k-60k g/mol | 11％w/w |
| 60k-200k g/mol | 27％w/w |
| 200k-1,100k g/mol | 35％w/w |
| 1,000k-1,600k g/mol | 4％w/w |
| >1,600k g/mol | 13％w/w |
| Content of sulfuric acid ester | 31％w/w |
| Ratio of acetyl to trehalose (%) | Less than 2 percent |
| Water content | Less than 10% w/w |
| pH of 0.1% w/v solution | 7 |
Fucoidan solution was prepared by dissolving 5g fucoidan in 50mL lactated ringer's injection USP, sterilizing with autoclaving and allowing the solution to cool to ambient temperature. A person with keloid characteristics (including symptoms of keloid characteristics, such as raised scars) is injected locally subcutaneously with a fucoidan solution at the site of the raised scar, 0.5mL of the fucoidan solution per subcutaneous injection. The severity of the raised scar was reduced after treatment with fucoidan solution.
Example 11: efficacy of fucoidan solution formulations for humans with keloid (keloid scar)
Fucoidan was extracted from brown algae Undaria pinnatifida and was determined to have the properties given in the following table using the analytical techniques described in the above examples:
| testing | Results |
| Appearance of the product | White to off-white powder |
| Total carbohydrate content | 61％w/w |
| Trehalose content | 52% w/w of total carbohydrates |
| Galactose content | 48% w/w of total carbohydrates |
| Other sugar monomers | Less than 1% w/w of total carbohydrate |
| Molecular weight of carbohydrate | |
| <5k g/mol | 8％w/w |
| 5k-20k g/mol | 2％w/w |
| 20k-60k g/mol | 11％w/w |
| 60k-200k g/mol | 27％w/w |
| 200k-1,100k g/mol | 35％w/w |
| 1,000k-1,600k g/mol | 4％w/w |
| >1,600k g/mol | 13％w/w |
| Content of sulfuric acid ester | 31％w/w |
| Ratio of acetyl to trehalose (%) | Less than 2 percent |
| Water content | Less than 10% w/w |
| pH of 0.1% w/v solution | 7 |
Fucoidan solution was prepared by dissolving 5g fucoidan in 50mL lactated ringer's injection USP, sterilizing with autoclaving and allowing the solution to cool to ambient temperature. A person with keloid (keloid scar) was injected locally subcutaneously with 0.5mL fucoidan solution at the site of the keloid scar, 0.5mL fucoidan solution per subcutaneous injection. The severity of the keloid scar was reduced after treatment with fucoidan solution.
Example 12: fucosan solution preparation for treating seborrheic dermatitis (dandruff)
Fucoidan was extracted from brown algae Undaria pinnatifida and was determined to have the properties given in the following table using the analytical techniques described in the above examples:
| testing | Results |
| Appearance of the product | White to off-white powder |
| Total carbohydrate content | 61％w/w |
| Trehalose content | 52% w/w of total carbohydrates |
| Galactose content | 48% w/w of total carbohydrates |
| Other sugar monomers | Less than 1% w/w of total carbohydrate |
| Molecular weight of carbohydrate | |
| <5k g/mol | 8％w/w |
| 5k-20k g/mol | 2％w/w |
| 20k-60k g/mol | 11％w/w |
| 60k-200k g/mol | 27％w/w |
| 200k-1,100k g/mol | 35％w/w |
| 1,000k-1,600k g/mol | 4％w/w |
| >1,600k g/mol | 13％w/w |
| Content of sulfuric acid ester | 31％w/w |
| Ratio of acetyl to trehalose (%) | Less than 2 percent |
| Water content | Less than 10% w/w |
| pH of 0.1% w/v solution | 7 |
Fucoidan solution was prepared by dissolving 5g fucoidan in 50mL lactated ringer's injection USP, sterilizing with autoclaving and allowing the solution to cool to ambient temperature. A person suffering from seborrheic dermatitis (dandruff) applied 5mL of fucoidan solution topically daily at the site of seborrheic dermatitis for 5 days. Seborrheic dermatitis was reduced after treatment with fucoidan solutions.
Example 13: efficacy of fucoidan solution formulations for persons suffering from contact dermatitis
Fucoidan was extracted from brown algae Undaria pinnatifida and was determined to have the properties given in the following table using the analytical techniques described in the above examples:
| testing | Results |
| Appearance of the product | White to off-white powder |
| Total carbohydrate content | 61％w/w |
| Trehalose content | 52% w/w of total carbohydrates |
| Galactose content | 48% w/w of total carbohydrates |
| Other sugar monomers | Less than 1% w/w of total carbohydrate |
| Molecular weight of carbohydrate | |
| <5k g/mol | 8％w/w |
| 5k-20k g/mol | 2％w/w |
| 20k-60k g/mol | 11％w/w |
| 60k-200k g/mol | 27％w/w |
| 200k-1,100k g/mol | 35％w/w |
| 1,000k-1,600k g/mol | 4％w/w |
| >1,600k g/mol | 13％w/w |
| Content of sulfuric acid ester | 31％w/w |
| Ratio of acetyl to trehalose (%) | Less than 2 percent |
| Water content | Less than 10% w/w |
| pH of 0.1% w/v solution | 7 |
Fucoidan cream was prepared by thoroughly mixing 5g fucoidan in 50mL Dermabase cream. Persons with contact dermatitis and the symptoms of red, itchy skin topically applied 5mL of fucoidan cream daily at the site of contact dermatitis for 5 days. Seborrheic dermatitis (including its symptoms) was reduced after treatment with the fucoidan cream.
Example 30: fucoisan solution preparation for treating acne rosacea
Fucoidan was extracted from brown algae Undaria pinnatifida and was determined to have the properties given in the following table using the analytical techniques described in the above examples:
| testing | Results |
| Appearance of the product | White to off-white powder |
| Total carbohydrate content | 61％w/w |
| Trehalose content | 52% w/w of total carbohydrates |
| Galactose content | 48% w/w of total carbohydrates |
| Other sugar monomers | Less than 1% w/w of total carbohydrate |
| Molecular weight of carbohydrate | |
| <5k g/mol | 8％w/w |
| 5k-20k g/mol | 2％w/w |
| 20k-60k g/mol | 11％w/w |
| 60k-200k g/mol | 27％w/w |
| 200k-1,100k g/mol | 35％w/w |
| 1,000k-1,600k g/mol | 4％w/w |
| >1,600k g/mol | 13％w/w |
| Content of sulfuric acid ester | 31％w/w |
| Ratio of acetyl to trehalose (%) | Less than 2 percent |
| Water content | Less than 10% w/w |
| pH of 0.1% w/v solution | 7 |
Fucoidan cream was prepared by thoroughly mixing 5g fucoidan in 50mL Dermabase cream. Persons with early stage rosacea and redness, itching skin symptoms topically applied 5mL fucoidan cream daily at the site of the rosacea for 5 days. Following treatment with the fucoidan cream, erythema and acne (including symptoms thereof) were reduced.
All embodiments, aspects, features, etc. can be mixed and matched, combined, and arranged in any desired manner, unless explicitly stated otherwise or otherwise apparent from the context. Unless otherwise indicated, the use of "or" includes "and vice versa, except in the claims. Non-limiting terms should not be construed as limiting unless expressly stated otherwise or the context clearly dictates otherwise. (e.g., "including," "having," and "containing" generally mean "including, without limitation"). The singular forms (including the claims) such as "a", "an", and "the" include plural referents unless the context clearly dictates otherwise.