[go: up one dir, main page]

WO2017192360A1 - Compositions et méthodes de traitement du prurit provoqué par une allergie - Google Patents

Compositions et méthodes de traitement du prurit provoqué par une allergie Download PDF

Info

Publication number
WO2017192360A1
WO2017192360A1 PCT/US2017/029961 US2017029961W WO2017192360A1 WO 2017192360 A1 WO2017192360 A1 WO 2017192360A1 US 2017029961 W US2017029961 W US 2017029961W WO 2017192360 A1 WO2017192360 A1 WO 2017192360A1
Authority
WO
WIPO (PCT)
Prior art keywords
thyex
kda
colostrum
thymus
itching
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/US2017/029961
Other languages
English (en)
Inventor
Ryan Ushijima
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
CMI RESEARCH MANAGEMENT LLC
Original Assignee
CMI RESEARCH MANAGEMENT LLC
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from US15/206,145 external-priority patent/US10842819B2/en
Application filed by CMI RESEARCH MANAGEMENT LLC filed Critical CMI RESEARCH MANAGEMENT LLC
Publication of WO2017192360A1 publication Critical patent/WO2017192360A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
    • A61K31/7048—Compounds having saccharide radicals and heterocyclic rings having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin, digitoxin or digoxin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/20—Milk; Whey; Colostrum
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/26—Lymph; Lymph nodes; Thymus; Spleen; Splenocytes; Thymocytes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines

Definitions

  • thymus extract compositions e.g., Thyex-l-Thyex 6A and Thyex-6B; Kyosenex® as described herein
  • colostrum for treating itching caused by allergy, including itching caused by inflammation related to the presence of mammalian parasites and/or viral infections, and in more particular veterinary aspects to treating itching caused by allergy, and itching associated with demodectic mange and stomatitis (e.g., gingivostomatitis) in the veterinary setting (e.g., treating canine, feline, bovine, equine, ovine, porcine, avian, etc., disorders).
  • demodectic mange and stomatitis e.g., gingivostomatitis
  • Combination or adjunctive therapies of the thymus extract compositions with colostrum are encompassed, as well as those using at least one additional anti-parasitic, anti -bacterial, anti-fungal, anti-viral agent, or homeopathic agent.
  • colostrum e.g., bovine colostrum
  • Itchy skin e.g., pruritus, dermatitis, eczema, psoriasis, dermatographism, etc.
  • Itchy skin is an irritating and uncontrollable sensation that makes you want to scratch to relieve the feeling.
  • the possible causes for itchiness range from internal illnesses, such as kidney or liver disease, to skin rashes, allergies, and dermatitis. Itching caused by allergy is a huge problem, particularly in the veterinary setting, and including that associated with
  • demodectic mange and stomatitis in the veterinary setting (e.g., treating canine, feline, bovine, equine (e.g., race horse), ovine, porcine, etc., disorders). Itching can be caused by viruses (chicken pox, measles) and other agents (e.g., fungus, mites, bedbugs, lice, pinworms, scabies, etc.).
  • Colostrum is the lacteal (mammary) secretion of mammals, produced immediately prior to birth and intended for feeding to the newborn. It is rich in protective and regulating factors necessary to promote health and vitality. It is nature's most concentrated source of biologically active components, particularly in the first, second and third milkings (e.g., with about 72 hours postpartum) It has existed since the first mammals roamed the planet and used by humans of various cultures for thousands of years. Cows are an ideal source of colostrum for at least the three following basic reasons: (i) nutritional, management and health improvements of dairy cows have led to greater colostrum production during the past 100 years. The calf s needs are always fulfilled first.
  • bovine colostrum is more potent than human colostrum in many regards. Newborn human infants have more competent immune function due to placental transfer of antibodies and various proteins/peptides which do not occur in ruminants (mammals with four chambered stomachs such as cows, goats and sheep). Because of this, these factors are concentrated and retained in the bovine colostrum for feeding to the calf immediately after birth; and (iii) the biologically active components are either identical or functionally identical regardless of the species of origin or use. In the first milking collected within six hours, over 80% of the albumin fraction is bioactive components produced in the bone marrow and circulatory system.
  • Thymus extracts Thymus extract compositions (e.g., Thyex-l-Thyex 6A and Thyex-6B; Kyosenex® are described herein.
  • Demodicosis also called demodectic mange or red mange
  • Demodex mites in the family Demodicidae
  • Demodex mites canis, for example, occurs naturally in the hair follicles of most dogs in low numbers around the face and other areas of the body. In most dogs, these mites never cause problems. However, in certain situations (e.g., underdeveloped or impaired immune system, intense stress, or malnutrition, the mites can reproduce rapidly, causing symptoms in sensitive dogs that range from mild irritation and hair loss on a small patch of skin to severe and widespread inflammation, secondary infection, and in rare cases can be a life-threatening condition.
  • Demodectic mange is transmitted from host to host through direct contact. Typically animals become infected through nursing from their mother. Demodex mites are host-adapted; there is no zoonotic potential in either canine or feline demodicosis. These mites (e,g., Demodex canis) thrive only on their specific hosts (dogs). The transmission of these mites from mother to pup, for example, is typical, but some individuals are sensitive to the mites due to a cellular immune deficiency, underlying disease, stress, or malnutrition, which can lead to the development of clinical demodectic mange.
  • Demodex cati causes follicular mange, similar to that seen in dogs, though it is much less common.
  • Demodex gatoi is a more superficial form of mange, causes an itchy skin condition, and is contagious amongst cats.
  • These diseases in humans are usually caused by Demodex folliculorum (not the same species affecting dogs) and are usually called demodicosis which may have a rosacea-like appearance. Common symptoms include hair loss, itching and inflammation. An association with pityriasis folliculorum has also been described.
  • Demodectic mange with secondary infection is treated with antibiotics and medicated shampoos.
  • Amitraz is a parasiticidal dip that is licensed for use in many countries (the only FDA approved treatment in the USA) for treating canine demodicosis. It is applied weekly or biweekly, for several weeks, until no mites can be detected by skin scrapings.
  • Demodectic mange in dogs can also be managed with avermectins, although there are few countries which license these drugs, which are given by mouth, daily, for this use.
  • Ivermectin is used most frequently; collie-like herding breeds often do not tolerate this drug due to a defect in the blood-brain barrier, though not all of them have this defect.
  • Other avermectin drugs that can be used include doramectin and milbemycin.
  • Cats with Demodex gatoi must be treated with weekly or bi-weekly sulfurated lime rinses. Demodex cati are treated similarly to canine demodicosis. Because of the possibility of the immune deficiency being an inherited trait, many veterinarians believe that all puppies with generalized demodex should be spayed or neutered and not reproduce. Females with generalized demodex should be spayed because the stress of the estrus cycle will often bring on a fresh wave of clinical signs. For really bad cases, the animal must be euthanized.
  • demodicosis forms include localized demodicosis, usually in juvenile dogs, occurs as isolated scaly bald patches, usually on the dog's face, but occasionally elsewhere. It is considered common and the majority of cases resolve with no treatment of any kind. Localized disease does not involve more than two body regions (One spot or two on the face and one spot or two on a leg would still qualify as localized even though the spots are not close together). Localized disease involves no more than four spots total on the dog. Generalized demodicosis can be very difficult, and requires persistent intervention and dedicated owners, as it is multifactorial and is often complicated by concurrent infections.
  • Demodectic Pododermatitis is a very resistant type, confined to the paws, and often accompanied by bacterial infections, as the foot is the last stronghold of the mite. Old English Sheepdogs and Shar Peis tend to get severe forms of this condition.
  • Sarcoptic mange which is caused by the deeply burrowing mite, Sarcoptes scabiei, is extremely contagious between dogs, and can be transmitted to people. It is the mange most people picture when the think of a "mangy dog". Dogs are highly pruritic, with progressive hair loss, reddened skin, and scabbing especially on ear flaps, eyes, elbows, feet, and chest. It is often difficult to detect on skin scrapings, so it often better to treat on clinical signs, even in the presence of a negative scraping.
  • Dermatophytosis Ringworm "dermatophytosis,” is a fungal infection affecting the skin, hair and occasionally nails of animals (and people). Three species of ringworm fungus most commonly affect cats and dogs. Microsporum canis, Trichophyton mentagrophytes and Microsporum gypseum. Gingivostomatitis. Stomatitis is inflammation in the mouth, and is also associated with irritation and itching. The term stomatitis refers to any inflammatory process affecting the mucous membranes of the mouth and lips, with or without oral ulceration.
  • the inflammation can be caused by conditions in the mouth itself, such as poor oral hygiene, dietary protein deficiency, poorly fitted dentures, or from mouth burns and scars from food or drinks, toxic plants, or by conditions that affect the entire body, such as medications, allergic reactions, radiation therapy, or infections.
  • gingivostomatitis refers to inflammation of the gingiva (i.e., gingivitis) and the mouth generally.
  • Cats with this chronic, painful inflammatory disease can be severely compromised, and medical treatment can cause adverse effects.
  • Affected cats exhibit a variety of clinical signs including partial to complete anorexia, ptyalism, halitosis, weight loss, abnormal swallowing, scratching, and oral pain.
  • Physical examination results show gingivitis, stomatitis, and possibly palatitis, glossitis, cheilitis, pharyngitis, and mandibular lymphadenopathy.
  • Oral inflammation is often extensive, and affected tissues are typically ulcerated, proliferative, and hyperemic. All breeds of cat have the potential to develop the disease, including domestic shorthaired cats. And cats can become affected at any age. Gingivostomatitis is not an infection but rather an inflammation.
  • the inflammatory lesions associated with feline gingivostomatitis are thought to be the result of a highly reactive immune system.
  • the specific antigen that the immune system is reacting to is not easily identified and is often unknown.
  • Antigens that may have a role in triggering the oral inflammation associated with feline gingivostomatitis include viral, food, or environmental antigens.
  • Autoimmunity may also be a component of the disease.
  • Herpetic gingivostomatitis is inflammation of the mouth caused by herpes simplex virus. Allergic gingivostomatitis, or allergic contact gingivostomatitis.
  • Treatment may include, for example, at least one of antibiotics, medicated mouth rinses, salt water, hydrogen peroxide, Xylocaine, antiviral agents, Acyclovir, fluid intake, good oral hygiene, gentle debridement of the mouth, etc.
  • Particular aspects provide methods for treating itching (e.g., allergic itching), including itching associated with gingivostomatitis and demodectic mange, comprising administration of proprietary thymus extracts (e.g., Thyex-1-6A and -6B; see working EXAMPLES 1-8 herein) used in combination with, or formulated with colostrum, where, for example, such administration provides for lessening, alleviating and/or halting and/or permanent reversal of itching symptoms, and for temporary and long-term relief for itching.
  • itching e.g., allergic itching
  • thymus extracts e.g., Thyex-1-6A and -6B; see working EXAMPLES 1-8 herein
  • the treatment with thymus extract compositions comprises administration of Kyosenex® prepared in accordance with working Examples 1 or 2 herein, and formulated with colostrum (e.g., bovine colostrum; first, second or third milking colostrums, or within 72 hours postpartum; preferably first milking colostrum is used for the formulations).
  • colostrum e.g., bovine colostrum; first, second or third milking colostrums, or within 72 hours postpartum; preferably first milking colostrum is used for the formulations.
  • the methods are practiced by administration of at least one of thymus extracts Thyex-1-6A and -6B (see working EXAMPLES 1-8 herein) used in combination with or formulated with colostrum, and therapeutic compositions comprising said Thyex / colostrum formulations.
  • the composition is Kyosenex® (a heat-treated, fractionated thymus extract composition comprising proteins or polypeptides having molecular weights in the range of 3.5 kDa to 30 kDa; see Applicant's U.S. Patent No.
  • preparing thymus extract compositions comprises: homogenizing thymus tissue; removing tissue debris therefrom to produce a supernatant; and concentrating and denaturing the supernatant to produce a clarified supernatant fraction.
  • the processes comprise further clarifying of the clarified supernatant by high-speed centrifugation at about 8,500 (g).
  • the processes further comprise filter sterilizing.
  • the pH and ionic strength of the resulting supernatant are physiologically compatible.
  • the pH and ionic strength of the resulting supernatant have values of about 7 and of about 0.85% (w/v), respectively.
  • the initial ratio of thymus tissue to aqueous homogenization fluid is about 350 g wet weight of thymus tissue to about 0.7 L of homogenization fluid.
  • the processes comprise further fractionating based on molecular weight to obtain a final fraction having proteins of about 3.5 to about 30 kDa.
  • Additional embodiments for preparing thymus extract compositions comprise: homogenizing thymus tissue; removing tissue debris therefrom to produce a supernatant; concentrating, denaturing, and clarifying the supernatant fraction; further concentrating the clarified supernatant fraction to produce a further concentrated fraction; fractionating the further concentrated fraction to remove molecules having a molecular weight less than about 3.5 kDa; and further fractionating based on molecular weight to obtain a final fraction having proteins of about 3.5 to about 30 kDa.
  • the processes further comprise adjusting the pH and/or ionic strength, of the final fraction to a physiological or therapeutically compatible value.
  • said adjusting is achieved by adding phosphate buffer and/or sodium chloride to produce a solution having a pH value of about 7, and/or an ionic strength of about 0.85% (w/v).
  • the processes further comprise filter sterilizing.
  • said sterilizing is achieved by using a 0.2 micron membrane filter.
  • the initial ratio of thymus tissue to aqueous homogenization fluid is about 350 g wet weight (about 400 ml) of thymus tissue to about 0.7 L of homogenization fluid.
  • compositions comprising: thymus extract compositions (Thyex-1-6A and -6B) formulated with colostrum produced in accordance with the above-described processes, and a pharmaceutically acceptable carrier.
  • thymus extract compositions Thiex-1-6A and -6B
  • colostrum produced in accordance with the above-described processes
  • a pharmaceutically acceptable carrier Preferably, the thymus extract composition Kyosenex® is used.
  • bovine colostrum; first, second or third milking colostrums, or within 72 hours postpartum are used; preferably first milking colostrum is used for the formulations.
  • the subject mammal treated includes, but is not limited to human, canine, feline, bovine, equine, ovine, and porcine.
  • Avian is also encompassed
  • the inventive Thyex / colostrum formulations are useful for reducing itching, comprising administering to a mammalian subject in need thereof a therapeutically-effective amount of a thymus extract composition (Thyex- 1-6 A and -6B) / colostrum formulation produced in accordance with the herein-described processes.
  • the inventive Thyex / colostrum formulations are useful for reducing itching, comprising administering to a mammalian subject in need thereof a therapeutically-effective amount of a thymus extract composition (Thyex- 1-6 A and -6B) / colostrum formulation produced in accordance with the herein-described processes.
  • the inventive Thyex / colostrum formulations are used in combination with administering of at least one additional anti-parasitic, anti-microbial agent (e.g., an antibiotic), antifungal agent, antiviral agent, or homeopathic agent.
  • additional anti-parasitic, anti-microbial agent e.g., an antibiotic
  • antifungal agent e.g., an antibiotic
  • antiviral agent e.g., an antibiotic
  • a subject e.g, mammalian subject
  • a therapeutically effective amount of a heat- treated, fractionated thymus extract composition in combination with or formulated with colostrum to provide for reducing itching in the subject, wherein a method for treating itching is afforded.
  • the methods can be applied, for example, to itching caused by allergy or itching caused by parasites, including situations wherein the parasite-mediated itching comprises itching in demodicosis (demodectic mange; e.g., canine demodicosis, including that caused by Sarcoptes scabiei; e.g., feline demodicosis caused by Demodex cati or by Demodex gatoi.
  • demodicosis demodicosis
  • the methods can also be applied to parasite-mediated itching comprising itching in stomatitis (e.g., wherein the stomatitis is gingivostomatitis) or itching itching in dermatophytosis (ring worm)).
  • administration of the thymus extract/colostrum combination or formulation is at least once or twice per day for at least two days, and may also be administered twice per day for at least two days, and then at least twice per week for at least one month, or may, for example be administered at least twice per day for at least a week.
  • the thymus extract composition comprises proteins or polypeptides having molecular weights in the range of 3.5 kDa to 30 kDa.
  • the thymus extract composition comprises proteins or polypeptides having molecular weights in the range of 3.5 kDa to 30 kDa, including within 5 to 14 kDa, and lacks proteins or polypeptides having molecular weights greater than 30 kDa and less than 3.5 kDa.
  • the colostrum is preferably bovine colostrum, for example, wherein the bovine colostrum comprises first milking colostrum, second milking colostrum or third milking colostrum (or within 72 hours postpartum).
  • administration of the thymus extract/colostrum combination or formulation may comprise treating with at least one additional anti-parasitic, anti -bacterial, anti-fungal, anti-viral agent, or homeopathic agent.
  • at least one antiparasitic agent comprises an avermectin (e.g., ivermectin, doramectin and/or n; preferably ivermectin).
  • the at least one anti-bacterial agent comprises an antibiotic, metronidazole, tinidazole, co-trimoxazole, cephamandole, ketoconazole, latamoxef, cefoperazone, amoxicillin, cefmenoxime, furazolidone, doxycycline and erythromycin.
  • the at least one anti-fungal agent comprises one or more of itraconazole, Terbinafine (Lamasil), clotrimazole (Lotrimin, Mycelex), fluconazole, ketoconazole (Spectazole), griseofulvin, econazole (Spectazole), miconazole, miconazole nitrate (Monistat-Derm), tolnaftate, thiabendazole (Tresaderm), lime-sulfur treatments, imidazoles, (eg., bifonazole, butoconazole, fenticonazole, isoconazole, oxiconazole, sertaconazole, sulconazole, and tioconazole), triazoles, fluconazole, itraconazole, isavuconazole, ravuconazole, posaconazole, voriconazole, and terconazole), thiazoles, abafung
  • the at least one anti-viral agent comprises at least one of combivir, boceprevir, abacavir, docosanol, aciclovir, didanosine, cidofovir, acyclovir, delavirdine, adefovir, amantadine, amprenavir, arbidol, darunavir atazanavir, atripla, zanamivir, or oseltamivir.
  • the at least one homeopathic agent comprises at least one of traumeel, homotox, SBGA (blue green algae), placenta, wobenzyme, spascupreel, IMM formula, yunnan paiyo, vitamin E, omega-3 fatty acids, semongrass oil, and cedar oil.
  • Administration of the thymus extract/colostrum combination or formulation is preferably oral, but may be at least one route selected from the group consisting of oral administration, injection, inhalation, topical application, and rectal administration.
  • compositions including therapeutic compositions, comprising a thymus extract in combination with colostum are provided, preferably wherein the thymus extract and the colostrum are bovine.
  • the thymus extract comprises proteins or polypeptides having molecular weights in the range of 3.5 kDa to 30 kDa
  • the bovine colostrum comprises first milking colostrum.
  • the thymus extract composition comprises proteins or polypeptides having molecular weights in the range of 3.5 kDa to 30 kDa, including within 5 to 14 kDa, and lacks proteins or polypeptides having molecular weights greater than 30 kDa and less than 3.5 kDa.
  • the combinations or formulations may alternately or additionally comprise second or third milking colostrums (preferably within 72 hours postpartum.
  • Figure 1 is a flow diagrammatic representation comprising an inventive Thy ex- 1 process embodiment for preparing a thymus extract composition.
  • Figure 2 is a flow diagrammatic representation comprising an inventive Thyex-2 process embodiment for preparing a thymus extract composition.
  • Figure 3 is a flow diagrammatic representation comprising an inventive Thyex-3 process embodiment for preparing a thymus extract composition.
  • Figure 4 is a flow diagrammatic representation comprising an inventive Thyex-4, -5 and -6 process embodiments for preparing a thymus extract composition.
  • Figure 5 shows day one of therapy for gingivostomatitis using Applicant's thymus extract composition.
  • Figure 6 shows thirty days of therapy for gingivostomatitis using Applicant's thymus extract composition (6 x 5 day cycles).
  • Figure 7 shows sixty days of therapy for gingivostomatitis using Applicant's thymus extract composition (12 x 5 day cycles).
  • Figure 8 shows arrival of demodectic mange patient at the animal control unit prior to Kyosenex® treatment.
  • Figure 9 shows initiation of Kyosenex® treatment; Day 1 (day 7 of ivermectin treatment).
  • Figure 10 shows day 8, beginning of second week of Kyosenex® treatment (3 injections total).
  • Figure 11 shows day 15, beginning of third week of Kyosenex® treatment (6 injections total).
  • Figure 12 shows day 22, beginning of fourth week of Kyosenex® treatment (9 injections total).
  • Figure 13 shows day 28, after discontinuation of Kyosenex® treatment (12 injections total). Note, in comparison with Figures 10 and 11, the regrowth of haircoat along the trunk.
  • Figure 14 shows patient in her permanent home, 2-months post-treatment with Kyosenex®.
  • aspects of the present invention provide methods for treating itching caused by allergy, and including itching cause by parasite-mediated inflammation, comprising administration to a mammalian subject in need thereof a therapeutically effective amount of a heat-treated, fractionated thymus extract composition in combination with colostrum, to provide for reducing itching, including allergic and parasite-mediated itching in the subject, wherein treating itching is afforded.
  • the heat-treated, fractionated thymus extract composition comprises proteins or polypeptides having molecular weights in the range of 3.5 kDa to 30 kDa.
  • the itching is allergic itching, including allergic itching relating to parasite-mediated inflammation in demodectic mange (demodicosis) caused by sensitivity to Demodex mites, and treating itching relating to demodicosis is afforded.
  • the demodicosis is that of canine demodicosis (e.g., that caused by Sarcoptes scabiei), or feline demodicosis (e.g., feline demodicosis caused by Demodex cati or by Demodex gatoi.
  • the parasite-mediated itching comprises itching in stomatitis, and wherein treating stomatitis related itching is afforded.
  • the stomatitis is gingivostomatitis, and treating gingivostomatitis itching is afforded.
  • the parasite-mediated itching comprises itching in dermatophytosis (ring worm), and treating dermatophytosis itching is afforded.
  • the heat-treated, fractionated thymus extract composition for example, formulated with colostrum, is administered at least once.
  • the heat-treated, fractionated thymus extract colostrum formulation c is administered at least once per day orally for at least 5 day to one month.
  • the heat- treated, fractionated thymus extract colostrum formulation is administered twice per day for at least 2 days, and then once per day for at least one week.
  • the heat- treated, fractionated thymus extract colostrum formulation is administered twice per week for at least one month.
  • the above methods further comprise treating with at least one additional anti-parasitic, anti -bacterial, anti-fungal, anti-viral agent, and/or with a homeopathic agent.
  • the at least one anti-parasitic agent comprises an avermectin (e.g., comprises at least one of ivermectin, doramectin and milbemycin); preferably, ivermectin.
  • the at least one anti -bacterial agent comprises an antibiotic.
  • the at least one anti-fungal agent comprises one or more of itraconazole, Terbinafine (Lamasil), clotrimazole (Lotrimin, Mycelex), Fluconazole, ketoconazole (Spectazole), griseofulvin, econazole (Spectazole), miconazole, miconazole nitrate (Monistat-Derm), tolnaftate, thiabendazole (Tresaderm), or lime-sulfur treatments.
  • the at least one anti-viral agent comprises acyclovir.
  • the at least one homeopathic agent comprises at least one of traumeel, homotox, SBGA (blue green algae), placenta, wobenzyme, spascupreel, EVIM formula, yunnan paiyo, vitamin E, omega-3 fatty acids, semongrass oil, and cedar oil.
  • inventive compositions are administered in combination with (or formulated with) the colostrum, and administering of at least one additional anti -parasitic, anti-mi crobial agent (e.g., an antibiotic), antifungal agent, antiviral agent, or homeopathic agent is also encompassed.
  • additional anti -parasitic, anti-mi crobial agent e.g., an antibiotic
  • antifungal agent e.g., an antibiotic
  • antiviral agent e.g., an antibiotic
  • the method further comprises separating molecules having molecular weights greater than about 30 kDa from the heat-treated, fractionated thymus extract composition, wherein a heat-treated, fractionated thymus extract composition comprising proteins or polypeptides having molecular weights in the range of about 3.5 kDa to about 30 kDa is provided.
  • the method comprises further clarifying of the clarified supernatant by high-speed centrifugation to produce a final clarified supernatant fraction, and optionally sterilizing the final clarified supernatant fraction to produce a sterile final clarified supernatant fraction. In certain aspects, sterilizing is achieved by passing the final clarified supernatant fraction through a membrane filter.
  • the initial ratio of thymus tissue to aqueous homogenization fluid is about 350 g wet weight of thymus tissue to about 0.7 L of homogenization fluid.
  • removing tissue debris from the aqueous thymus homogenate is achieved by a combination of low-speed centrifugation and crude filtration.
  • heat denaturing and clarifying of the primary supernatant is achieved by heat denaturation, followed by low-speed centrifugation and crude filtration to remove particulate matter.
  • the methods further comprise lyophilization of the final clarified supernatant fraction. Preferably no steps involving exogenously added protease digestion, or extraction with organic solvents are used.
  • Additional particular aspects provide a method for preparing a thymus extract composition, comprising: homogenizing thymus tissue with aqueous homogenization fluid to produce an aqueous thymus homogenate; removing tissue debris from the aqueous thymus homogenate to produce a primary supernatant; heat denaturing the primary supernatant, and clarifying the denatured primary supernatant by use of at least one of low- speed centrifugation and filtration to produce an intermediate clarified supernatant; concentrating the intermediate clarified supernatant to produce a concentrated intermediate fraction; and separating molecules having molecular weights less than about 3.5 kDa from the concentrated intermediate fraction, wherein a heat-treated, fractionated thymus extract composition lacking proteins or polypeptides having molecular weights less than about 3.5 kDa is provided.
  • the method further comprises separating molecules having molecular weights greater than about 30 kDa from the heat-treated, fractionated thymus extract composition, wherein a heat-treated, fractionated thymus extract composition comprising proteins or polypeptides having molecular weights in the range of about 3.5 kDa to about 30 kDa is provided. Certain embodiments further comprise clarifying of the concentrated intermediate fraction by high-speed centrifugation to produce a final clarified supernatant fraction.
  • Particular aspects further comprise adjusting at least one of the pH or ionic strength of the fraction having proteins or polypeptides of molecular weight of about 3.5 to about 30 kDa to a physiological or therapeutically compatible value, to produce a pH- or ionic strength-adjusted fraction, and in certain aspects, adjusting at least one of the pH or ionic strength to a physiological or therapeutically compatible value is achieved by adding phosphate buffer or sodium chloride to produce a fraction having at least one of a pH value of about 7 or an ionic strength of about 0.85% w/v.
  • Certain embodiments further comprise sterilizing the pH-, or ionic strength-adjusted fraction to produce a sterile pH-, or ionic strength-adjusted fraction, and in particular aspects, sterilizing is achieved by passing the fraction through a membrane filter.
  • the initial ratio of thymus tissue to aqueous homogenization fluid is about 350 g wet weight of thymus tissue to about 0.7 L of homogenization fluid.
  • removing tissue debris from the aqueous thymus homogenate is achieved by a combination of low-speed centrifugation and crude filtration.
  • heat denaturing and clarifying of the primary supernatant is achieved by heat denaturation, followed by low-speed centrifugation and crude filtration to remove particulate matter.
  • concentrating the intermediate supernatant involves concentrating and fractionating, wherein the concentrating and fractionating is achieved by adding ammonium sulfate to the intermediate clarified supernatant, followed by low-speed centrifugation and suspension of the resulting ammonium sulfate pellet in an aqueous solution to provide a concentrated intermediate fraction.
  • separating molecules having molecular weights less than about 3.5 kDa from the concentrated intermediate fraction comprises dialysis of the concentrated intermediate fraction, followed by high-speed centrifugation to remove particulate matter, to provide for a clarified concentrated intermediate fraction lacking proteins or polypeptides having molecular weights less than about 3.5 kDa.
  • separating molecules having molecular weights greater than about 30 kDa from the heat-treated, fractionated thymus extract composition is achieved by passing the clarified concentrated intermediate fraction lacking proteins or polypeptides having molecular weights less than about 3.5 kDa consecutively through a first and a second membrane filter having exclusion limits of about 100 and about 30 kDa, respectively, and collecting the filtrate.
  • Particular aspects further comprise lyophilization of the heat-treated, fractionated thymus extract composition comprising proteins or polypeptides having molecular weights in the range of about 3.5 kDa to about 30 kDa.
  • compositions or pharmaceutical composition comprising a thymus extract composition, produced in accordance with the methods recited herein— in combination with Colostrum.
  • the macrophage stimulating agent comprises at least one of beta glucan, polysaccharides, toxoid vaccines, and Staph lysate vaccine, immune complexes, compliment components, lymphokinesm, tuftsin, lipopolysaccharides (LPS), muramyl dipeptide, physiologic cation complexing agents, pyran copolymers, polycarboxylates, ionphores, Quadrol ( ⁇ , ⁇ , ⁇ ', ⁇ '- tetrakis(2-hydroxypropyl)ethlenediamine), and macrophage stimlulating peptides.
  • the beta glucan comprises beta 1,3 glucan.
  • Further aspects provide a method for immunostimulation or immunoregulation, comprising administering to a mammalian subject in need thereof a therapeutically-effective amount of a thymus extract / colostrum formulation produced in accordance with the methods recited herein, wherein at least one of immunostimulation or immunoregulation is afforded.
  • Yet further aspects provide a method for endocrine modulation, comprising administering to a mammalian subject in need thereof a therapeutically-effective amount of a thymus extract / colostrum formulation produced in accordance with the methods recited herein, wherein endocrine modulation is afforded.
  • Thymus extract or thymus extract composition, refers to a composition produced in accordance with one or more of the Thyex-1, -2, -3, -4, -5, -6A and -6B processes disclosed herein.
  • Kyosenex® refers to a heat-treated, fractionated thymus extract composition comprising proteins or polypeptides having molecular weights in the range of 3.5 kDa to 30 kDa (e.g., see Applicant's U.S. Patent No. 8,609,824), preferably prepared as described under Examples 1 or 2 herein, although Kyosenex® can be prepared in accordance with the methods described under any of Examples 1-8 herein, providing a heat- treated, fractionated thymus extract composition comprising proteins or polypeptides having molecular weights in the range of 3.5 kDa to 30 kDa.
  • Kyosenex® is made according to the method described under Examples 1 or 2. Typically, it is provided in a sterile vial containing 3-4 mg of lyophilized thymus extract.
  • Colostrum refers to milking colostrum, and in particular aspects bovine colostrum is used.
  • first milking bovine colostrum is used (e.g., "First Milking Bovine ColostrumTM” from Immuno-Dynamics, Fennimore, WI; sold as "ID-1" optionally with methyl paraben and/or propyl paraben as preservative).
  • second, third or even further milkings are used.
  • second and/or third milking colostrum is used (e.g., within 72 hours postpartum).
  • first milking colostrum is used.
  • first milking bovine colostrum is used.
  • bovine colostrum is used to formulate the thymus extract for administration, and in certain embodiments, first milking bovine colostrum is used (e.g., "First Milking Bovine ColostrumTM" from Immuno-Dynamics, Fennimore, WI; sold as "ID-1" optionally with methyl paraben and/or propyl paraben as preservative).
  • Kyosenex® PRIME is formulated (formulated for Applicant by Immuno-Dynamic & URL Laboratories, Fennimore, WI USA, using ID-1 serum ("Bovine IgG Colostrum Serum”)) using 66.6 ug of Kyosenex/ml colostrum and administered in 7/10 ml (0.7 ml) per spray or dropwise, twice daily, 1 spray for each 15 lbs until resolution of condition, and then twice a week thereafter for maintenance.
  • ID-1 serum Bovine IgG Colostrum Serum
  • Animals as used herein for treatment of subjects refers to, but are not limited to chicken, duck, fish, hamster, rat, guinea pig, human, canine, feline, bovine, equine (e.g., race horse), avian, ovine, goat and porcine. Preferred animals are mammals.
  • Anti-microbial agent means an agent with, for example, antibacterial, antifungal or antiviral activity, including, but not limited to: plant extracts ⁇ e.g., Houttuynia cordata extracts); antibiotics, such as ⁇ -lactam antibiotics, erythromycin compounds, Tetracycline compounds, aminoglycoside antibiotics, cephalosporin compounds, anthracycline compounds, phleomycin group antibiotics, sulfonamide compounds, macrolide antibiotics ⁇ e.g., tylosin, desmycosin, macrocin, and lactenocin), quinolone and quinolonyl compounds ⁇ e.g., quinolonyl lactams and quinolone thioureas, and carbacephem- and carbapenem- quinolones) carbapenem compounds, along with those antibiotic agents more commonly used in the swine industry, such as lankacidin-group antibiotics and derivatives,
  • “Crude filtration” or “coarse filtration” means filtering a solution having particulate, precipitated or flocculent suspended material through, e.g., one or more layers of standard cheese cloth, or other sieving device ⁇ e.g., screen, strainer, colander, etc.), to remove said material.
  • Low-speed centrifugation means centrifugation at about 3,500 x g ( ⁇ 5% or ⁇ 10%) for about 5-10 minutes ( ⁇ 5% or ⁇ 10%), or an equivalent sedimentation protocol thereof.
  • High-speed centrifugation means centrifugation at about 8,500 x g ( ⁇ 5% or ⁇ 10%)) for about 10 minutes ( ⁇ 5% or ⁇ 10%), or the equivalent sedimentation protocol thereof.
  • "Clarifying,” or clarification of a supernatant fraction means removing particulate matter (e.g., precipitates, bacteria) from a solution containing such particulate matter through the use of standard separation techniques, such as low- or high-speed centrifugation (as defined above) or filtration.
  • the phrase "less than about 3.5 kDa” as used herein refers to less than 3.5 kDa, or less than a molecular weight that varies by ⁇ 5% or ⁇ 10% therefrom.
  • the phrase “proteins or polypeptides of molecular weight of about 3.5 to about 30 kDa” as used herein refers to proteins or polypeptides in a molecular weight ranged from 3.5 kDa, or from a molecular weight that varies by ⁇ 5% or ⁇ 10% therefrom, to 30 kDa, or to a molecular weight that varies by ⁇ 5% or ⁇ 10% therefrom.
  • a pH value of about 7, or an ionic strength of about 0.85% w/v refers to a pH of 7 or a pH that varies by ⁇ 5% or ⁇ 10%) therefrom, and/or an ionic strength of 0.85% w/v, or an ionic strength that varies by ⁇ 5% or ⁇ 10% therefrom.
  • Vaccine is defined herein in its broad sense to refer to any type of biological agent, administrable for the purpose of priming, enabling or enhancing an immune response against in an animal inoculated with the vaccine.
  • Particular embodiments of the present invention provide novel processes for preparing therapeutically useful extracts (Thyex-1-6A and -6B) of thymus tissue.
  • the inventive processes are readily distinguishable from other known processes for preparing thymus extracts (e.g., Goldstein & White, Contemp. Topics in Immunobiology, p339, 1973; Bergesi et al., Folia Allergol. Immunol. Clin. 21 :201, 1977; Hooper et al., "The purification and properties of bovine thymosin," Ann. NY Acad. Sci. 249: 125, 1975; U.S.
  • the instant processes comprise steps to optimize protein compositions for therapeutic use of.
  • Thyex-1-6A and -6B are designed to provide therapeutic compositions, and include ammonium sulfate precipitation/fractionation and/or lyophilization steps, respectively, to facilitate optimal protein concentration and fractionation.
  • the Thyex-3 process embodiment lacks an ammonium sulfate or lyophilization step, but provides for a sufficiently-concentrated composition by reusing (and thereby augmenting) an initial tissue homogenization supernatant fraction as homogenization fluid to homogenize additional tissue.
  • the resulting Thyex-3 composition is less refined relative to those of Thyex-1 and Thyex-2, but is nonetheless suitably concentrated and formulated for efficacious delivery.
  • Thyex 6A and Thyex 6B process embodiments described below are designed to provide therapeutic compositions suitable for delivery as a topical ointment or by injection or inhalation, and include ammonium sulfate precipitation/fractionation steps.
  • Thyex 5 is prepared from a similar process but is less refined (less fractionated) than Thyex 6A or Thyex 6B and is optimally mixed with an amount of an extracted lyophilized herbal source composition, and administered orally in filled gelatin capsules.
  • the Thyex 4 process embodiment lacks ammonium sulfate precipitation step but comprises lyophilization to provide for a sufficiently-concentrated composition.
  • the resulting Thyex 4 composition is less refined in relative to those of Thyex 5 or Thyex 6A or 6B, but is nonetheless suitably concentrated and formulated for efficacious oral deliver in both animals and humans.
  • Particular specific aspects provide a method for preparing a thymus extract composition, comprising: homogenizing thymus tissue with aqueous homogenization fluid to produce an aqueous thymus homogenate; removing tissue debris from the aqueous thymus homogenate to produce a primary supernatant; and heat denaturing and clarifying the primary supernatant to produce a clarified supernatant.
  • the method further comprises further clarifying of the clarified supernatant by high-speed centrifugation to produce a final clarified supernatant fraction.
  • the method further comprises sterilizing the final clarified supernatant fraction to produce a sterile final clarified supernatant fraction.
  • sterilizing is achieved by passing the final clarified supernatant fraction through a membrane filter.
  • the initial ratio of thymus tissue to aqueous homogenization fluid is about 350 g wet weight of thymus tissue to about 0.7 L of homogenization fluid.
  • removing tissue debris from the aqueous thymus homogenate is achieved by a combination of low-speed centrifugation and crude filtration.
  • heat denaturing and clarifying of the primary supernatant is achieved by heat denaturation, followed by low-speed centrifugation and crude filtration to remove particulate matter.
  • the method further comprises lyophilization of the final clarified supernatant fraction.
  • Additional aspects provide a method for preparing a thymus extract composition, comprising: homogenizing thymus tissue with aqueous homogenization fluid to produce an aqueous thymus homogenate; removing tissue debris from the aqueous thymus homogenate to produce a primary supernatant; heat denaturing and clarifying the primary supernatant to produce an intermediate supernatant; and concentrating the intermediate supernatant to produce a concentrated intermediate fraction.
  • the method further comprises further clarifying of the concentrated intermediate fraction by high-speed centrifugation to produce a final clarified supernatant fraction.
  • the method further comprises fractionating the final clarified supernatant fraction to remove molecules having a molecular weight less than about 3.5 kDa to produce a fractionated intermediate fraction.
  • the method further comprises fractionating the fractionated intermediate fraction, based on molecular weight, to obtain a fraction having proteins of about 3.5 to about 30 kDa.
  • the method further comprises adjusting at least one of the pH or ionic strength of the fraction having proteins of about 3.5 to about 30 kDa to a physiological or therapeutically compatible value, to produce a pH- or ionic strength-adjusted fraction.
  • adjusting at least one of the pH or ionic strength to a physiological or therapeutically compatible value is achieved by adding phosphate buffer or sodium chloride to produce a fraction having at least one of a pH value of about 7 or an ionic strength of about 0.85% w/v.
  • the method further comprises sterilizing the pH-, or ionic strength-adjusted fraction to produce a sterile pH-, or ionic strength-adjusted fraction.
  • sterilizing is achieved by passing the fraction through a membrane filter.
  • the initial ratio of thymus tissue to aqueous homogenization fluid is about 350 g wet weight of thymus tissue to about 0.7 L of homogenization fluid.
  • removing tissue debris from the aqueous thymus homogenate is achieved by a combination of low-speed centrifugation and crude filtration.
  • heat denaturing and clarifying of the secondary supernatant is achieved by heat denaturation, followed by low-speed centrifugation and crude filtration to remove particulate matter.
  • concentrating the intermediate supernatant involves concentrating and fractionating, and wherein the concentrating and fractionating is achieved by adding ammonium sulfate to the intermediate supernatant, followed by low-speed centrifugation and suspension of the resulting ammonium sulfate pellet in an aqueous solution.
  • fractionating the concentrated intermediate fraction to remove molecules having a molecular weight less than about 3.5 kDa is achieved by dialysis of the concentrated intermediate fraction, followed by high-speed centrifugation to remove particulate matter.
  • fractionating the fractionated intermediate fraction, based on molecular weight is achieved by passing the fractionated intermediate fraction consecutively through a first and a second membrane filter having exclusion limits of about 100 and about 30 kDa, respectively, and collecting the filtrate.
  • the method further comprises lyophilization of the fraction having proteins of about 3.5 to about 30 kDa.
  • Particular specific aspects provide a process for preparing a thymus extract composition, comprising: homogenizing thymus tissue with aqueous homogenization fluid to produce an aqueous thymus homogenate; removing tissue debris from the aqueous thymus homogenate to produce a primary supernatant; concentrating the primary supernatant to produce a secondary supernatant; and denaturing and clarifying the secondary supernatant to produce a clarified supernatant.
  • the method further comprises further clarifying of the clarified supernatant by high-speed centrifugation to produce a final clarified supernatant fraction.
  • the method further comprises sterilizing the final clarified supernatant fraction to produce a sterile final clarified supernatant fraction.
  • sterilizing is achieved by passing the final clarified supernatant fraction through a membrane filter.
  • the initial ratio of thymus tissue to aqueous homogenization fluid is about 300 g wet weight, or about 340 ml wet volume, of thymus tissue to about 0.8 L of homogenization fluid.
  • removing tissue debris from the aqueous thymus homogenate is achieved by a combination of low-speed centrifugation and crude filtration.
  • concentrating the primary supernatant is achieved by repeating (a) and (b) using the primary supernatant, in place of the aqueous homogenization fluid, for homogenizing additional thymus tissue.
  • denaturing and clarifying of the secondary supernatant is achieved by heat denaturation, followed by low-speed centrifugation and crude filtration to remove particulate matter.
  • Additional specific aspects provide a method for preparing a thymus extract composition, comprising: homogenizing thymus tissue with aqueous homogenization fluid to produce an aqueous thymus homogenate; removing tissue debris from the aqueous thymus homogenate to produce a primary supernatant; concentrating the primary supernatant to produce a secondary supernatant; denaturing and clarifying the secondary supernatant to produce an intermediate supernatant; concentrating the intermediate supernatant to produce a concentrated intermediate fraction; fractionating the concentrated intermediate fraction to remove molecules having a molecular weight less than about 3.5 kDa to produce a fractionated intermediate fraction; and fractionating the fractionated intermediate fraction, based on molecular weight, to obtain a fraction having proteins of about 3.5 to about 30 kDa.
  • the method further comprises adjusting at least one of the pH or ionic strength of the fraction having proteins of about 3.5 to about 30 kDa to a physiological or therapeutically compatible value, to produce a pH- or ionic strength-adjusted fraction.
  • adjusting at least one of the pH or ionic strength to a physiological or therapeutically compatible value is achieved by adding phosphate buffer or sodium chloride to produce a fraction having at least one of a pH value of about 7 or an ionic strength of about 0.85% w/v.
  • the method further comprises sterilizing the pH-, or ionic strength-adjusted fraction to produce a sterile pH-, or ionic strength-adjusted fraction.
  • sterilizing is achieved by passing the fraction through a membrane filter.
  • the initial ratio of thymus tissue to aqueous homogenization fluid is about 350 g wet weight of thymus tissue to about 0.7 L of homogenization fluid.
  • removing tissue debris from the aqueous thymus homogenate is achieved by a combination of low-speed centrifugation and crude filtration.
  • concentrating the primary supernatant is achieved by repeating (a) and (b) using the primary supernatant, in place of the aqueous homogenization fluid, for homogenizing additional thymus tissue.
  • denaturing and clarifying of the secondary supernatant is achieved by heat denaturation, followed by low-speed centrifugation and crude filtration to remove particulate matter.
  • the intermediate supernatant is concentrated, wherein concentrating is achieved by lyophilizing the intermediate supernatant either to complete dryness followed by aqueous resuspension to about 500 ml/13.6 kg (30 lbs.) original wet tissue, or to a volume of about 10% of its original volume.
  • concentrating the intermediate supernatant involves concentrating and fractionating, and wherein the concentrating and fractionating is achieved by adding ammonium sulfate to the intermediate supernatant, followed by low-speed centrifugation and suspension of the resulting ammonium sulfate pellet in an aqueous solution.
  • fractionating the concentrated intermediate fraction to remove molecules having a molecular weight less than about 3.5 kDa is achieved by dialysis of the concentrated intermediate fraction, followed by high- speed centrifugation to remove particulate matter.
  • fractionating the fractionated intermediate fraction is achieved by passing the fractionated intermediate fraction consecutively through a first and a second membrane filter having exclusion limits of about 100 and about 30 kDa, respectively, and collecting the filtrate.
  • compositions comprising a thymus extract composition produced in accordance with one or more of the processes disclosed herein.
  • treating refers to, and includes, reversing, alleviating, inhibiting the progress of, or preventing a disease, disorder or condition, or one or more symptoms thereof; and "treatment” and “therapeutically” refer to the act of treating, as defined herein.
  • a “therapeutically-effective amount” is any amount of any of the compounds utilized in the course of practicing the invention provided herein that is sufficient to reverse, alleviate, inhibit the progress of, or prevent a disease, disorder or condition, or one or more symptoms thereof.
  • the methods comprise administration of a composition comprising at least one of Thyex-1-6A and -6B, as defined herein, and formulated with colostrum, including optionally in combination with (e.g., adjunctive therapy), for example, with administration of an anti-parasitic agent, anti-viral agent, antibacterial agent, anti-fungal agent, and/or with a macrophage stimulating agent.
  • a composition comprising at least one of Thyex-1-6A and -6B, as defined herein, and formulated with colostrum, including optionally in combination with (e.g., adjunctive therapy), for example, with administration of an anti-parasitic agent, anti-viral agent, antibacterial agent, anti-fungal agent, and/or with a macrophage stimulating agent.
  • a polysaccharide is used as preferred macrophage stimulating agent.
  • the macrophage stimulating agent comprises a beta glucan.
  • the beta glucan comprises at least one linkage selected from the group consisting of beta: 1,3; 1,4; and 1,6 glucan linkages.
  • the linkage is that of beta 1,3 glucan.
  • the inventive Thyex compositions are used in adjunctive therapies with extracts of at least one of: Paresis crepe (aka cauliflower mushroom or hanabaritake) preparations comprising beta 1-3 glucan; Lentinula edodes (shitake; e.g., alkaline digest according to the procedure reported by Ohno et al. (Biol. Phar. Bull. 23 866-872, 2000), comprises beta 1-3 glucan and chitin; Astralagas membranaceus; Scutellaria baicalensis; Lilium longi forum (aka Easter lily); and Houttuynia cordata extracts.
  • Paresis crepe aka cauliflower mushroom or hanabaritake
  • Lentinula edodes e.g., alkaline digest according to the procedure reported by Ohno et al. (Biol. Phar. Bull. 23 866-872, 2000), comprises beta 1-3 glucan and chitin; Astralagas
  • compositions comprising a thymus extract composition produced in accordance with one or more of the processes disclosed herein.
  • Combination therapies are also encompassed by aspects of the present invention.
  • the inventive methods may further comprise administration of a therapeutically-effective amount of one or more anti-microbial agents, such as anti-parasitic, anti-viral agents, anti-bacterial agents, anti-fungal agents.
  • anti-viral agents include but are not limited to: combivir, boceprevir, abacavir, docosanol, aciclovir, didanosine, cidofovir, acyclovir, delavirdine, adefovir, amantadine, amprenavir, arbidol, darunavir atazanavir, atripla, zanamivir, and oseltamivir.
  • anti -bacterial agents include but are not limited to: metronidazole, tinidazole, co- trimoxazole, cephamandole, ketoconazole, latamoxef, cefoperazone, amoxicillin, cefmenoxime, furazolidone, doxycycline and erythromycin.
  • anti-funal agents include but are not limited to: imidazoles, (e.g., miconazole, ketoconazole, clotrimazole, econazole, bifonazole, butoconazole, fenticonazole, isoconazole, oxiconazole, sertaconazole, sulconazole, and tioconazole), triazoles (e.g., fluconazole, itraconazole, isavuconazole, ravuconazole, posaconazole, voriconazole, and terconazole), thiazoles (e.g., abafungin), allylamines (e.g., terbinafine, amorolfine, naftifine, and butenafine), and echinocandins (e.g., anidulafungin, caspofungin, and micafungin).
  • imidazoles e.g., micon
  • Thyex compositions have substantial utility for treating stomatitis (e.g., gingivostomatitis).
  • stomatitis e.g., gingivostomatitis
  • a thymic extract, Kyosenex® was used in management of a protracted case of feline gingivostomatitis.
  • the patient responded dramatically, especially in the caudal portion of the oral cavity.
  • the condition recurred upon stopping therapy and improved again on reinstituting the agent. Quality of life was greatly improved.
  • Preferred embodiments relates to a method for treating stomatitis (e.g., gingivostomatitis), comprising administering to a mammalian subject in need thereof an effective amount of a thymus extract composition produced in accordance with the methods recited herein, wherein effects of stomatitis (e.g., gingivostomatitis related inflammation) alleviated.
  • stomatitis e.g., gingivostomatitis
  • the mammalian subject in need of treatment includes but is not limited to canine, feline, bovine, porcine, equine, ovine, and other large animals.
  • the method for treating stomatitis includes veterinary applications (e.g., canine and feline).
  • veterinary applications e.g., canine and feline.
  • Thyex compositions have substantial utility for treating demodectic mange.
  • a thymic extract, Kyosenex® was used in management of a protracted case of canine demodectic mange. The patient responded dramatically. At one-year post treatment, there has been no recurrence of the condition.
  • compositions are Compositions:
  • Thyex-1, -2, -3, -4, -5, -6A and -6B composition embodiments are produced in accordance with the corresponding Thyex- 1-6 A and -6B processes (Working EXAMPLES 1-8).
  • Thyex-1-3 processes provide exemplary process embodiments used for preparing Thyex-1-3, produced in accordance therewith suitable for oral delivery.
  • Thyex-1-3 are lyophilized, stored at ambient temperature and reconstituted with sterile water prior to use.
  • Steps (l)-(l l) of EXAMPLE 4 comprise a process embodiment for producing Thyex-4 (step (12) relates to storage), suitable for oral delivery.
  • Thyex-4 is lyophilized, stored at ambient temperature and reconstituted with sterile water prior to use.
  • Steps (1)-(13) EXAMPLE 5 comprise a process embodiment for producing Thyex-5 (step (14) relates to storage), suitable for oral delivery.
  • Thyex-5 is lyophilized, stored at ambient temperature and reconstituted with sterile water prior to use.
  • Thyex-6A process provides an exemplary process embodiment used for preparing Thyex-6A produced in accordance therewith suitable for oral delivery, or delivery as a topical ointment or by injection or inhalation.
  • Thyex-6A is lyophilized, stored at ambient temperature and reconstituted with sterile water prior to use.
  • Thyex-6B process provides an exemplary process embodiment used for preparing Thyex-6B produced in accordance therewith suitable for oral delivery, or delivery as a topical ointment or by injection or inhalation.
  • Thyex-6B was lyophilized, stored at ambient temperature and reconstituted with sterile water prior to use.
  • a therapeutically-effective dose of a composition of the present invention refers to that amount of the composition sufficient to prevent or inhibit the effects of the treated condition, or to that amount sufficient to enhance the efficacy of adjunctive regimens. This amount may vary somewhat among subjects, but are nonetheless reasonably determined by one of ordinary skill within the art in view of the many art-recognized symptoms associated with the treated conditions.
  • Therapeutically-effective doses of the disclosed compositions are administered alone or in combination with other therapeutic agents, such as macrophage stimulating agents, anti-microbial agents (e.g., antiviral, antifungal or antibacterial agents), or are administered as adjunctive therapy in combination with administration of other treatment regimens.
  • therapeutic agents such as macrophage stimulating agents, anti-microbial agents (e.g., antiviral, antifungal or antibacterial agents)
  • anti-microbial agents e.g., antiviral, antifungal or antibacterial agents
  • the Thyex compositions are standardized at a protein concentration about 2 mg/ml. Lyophilized compositions may be suspended, for example, using physiological saline with sodium chloride of 0.85-0.9%.
  • the daily dose range for Thyex administration by injection is from about 0.05 mg/kg to about 1 mg/kg. More preferably, the dose range for Thyex administration by injection is from about 0.05 mg/kg to about 0.5 mg/kg. Even more preferably, the dose range for Thyex administration by injection is from about 0.1 mg/kg to about 0.4 mg/kg. Most preferably, the dose range for Thyex administration by injection is from about 0.2 mg/kg to about 0.3 mg/kg.
  • the daily dose range for Thyex oral administration is from about 1 mg/kg to about 20 mg/kg. More preferably, the dose range for Thyex oral administration is from about 1 mg/kg to about 10 mg/kg. Even more preferably, the dose range for Thyex oral administration is from about 3 mg/kg to about 9 mg/kg. Most preferably, the dose range for Thyex oral administration is from about 5 mg/kg to about 8 mg/kg.
  • the daily dose range for adjunctive administration of beta glucan can be determined by routine optimization by one of ordinary skill in the art.
  • the daily dose range for adjunctive administration of the polysaccharide extract e.g., consisting of about 70% beta 1-3 glucan and 30% tissue proteins
  • the daily dose range for adjunctive administration of the polysaccharide extract will be about 300 to about 500 mg per day for a typical patient (e.g., or about 0.5 mg/kg to 15 mg/kg).
  • the daily dose will be about 300 mg per day (e.g., or about 0.5 mg/kg to 2.0 mg/kg) for a typical patient.
  • Antibiotic dosages were those of the label, according to the particular antibiotic used.
  • Doses of antifungal, antiviral, antiparasitic are according to those of the label, according to the particular agent used.
  • typical doses for ivermectin are: 6 ug/kg for heartworm prevention; 300 ug/kg for treatment of sarcoptic mange; and 400-600 ug/kg for treatment of demodectic mange.
  • Dipping - Paramite dip (a discontinued organophosphate), and Lime-Sulfur dips were mainstays of treatment for Sarcoptes, but of very limited value in Demodecosis. Goodwinol ointment- used for many decades, since approximately 10% of localized demodicosis cases will progress to generalized demodicosis, often accompanied by lymphadenopathy.
  • Moxidectin (Advantage Multi®) can be used to treat demodicosis and is often effective if used weekly.
  • Amitraz (Mitaban) Dip is an old therapy, with best efficacy at double strength and applied weekly. It can be quite toxic, particularly in small dogs. Preventic (amitraz) collars have had some success, but they must be changed often (about every 4 weeks).
  • Milbemycin oxime can be an effective, expensive approach to generalized demodicosis (0.52 to 3.8 mg/kg of body weight, q 24 hr). May be used in dogs with genetic sensitivity to Ivermectin (herding breeds, primarily, carrying the MDR gene). Some dogs require concurrent dipping.
  • Thyex-1-6A and-6B in combination with, or formulated with colostrum have substantial utility in methods for treatment of itching associated with various mammalian conditions including, but not limited to itching relating to allergy, demodectic mange and stomatitis (e.g., gingivostomatitis), comprising administration (e.g., oral) of said compositions.
  • stomatitis e.g., gingivostomatitis
  • Thy ex compositions and colostrum formulations thereof of the present invention are preferably formulated in aqueous solutions with physiologically compatible buffered saline (e.g., phosphate buffered standard physiological saline; 0.85% NaCl).
  • physiologically compatible buffered saline e.g., phosphate buffered standard physiological saline; 0.85% NaCl.
  • the pharmaceutical Thyex compositions and colostrum formulations thereof of the present invention may take the form of, for example, liquids, gels, syrups, slurries, and the like, prepared by conventional means with pharmaceutically acceptable excipients such as: binding agents (e.g., pre-gelatinized maize starch, wheat starch, rice starch, potato starch, gelatin, gum tragacanth, methyl cellulose, hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, and/or polyvinylpyrrolidone (PVP)); fillers (e.g., lactose, sucrose, mannitol, or sorbitol, microcrystalline cellulose or calcium hydrogen phosphate); lubricants (e.g., magnesium stearate, talc or silica); disintegrants (e.g., potato starch, sodium starch glycolate, cross- linked polyvinyl pyrrolidone, agar, or alginic acid or a salt thereof
  • Such liquid preparations are prepared by conventional means with pharmaceutically acceptable additives such as suspending agents (e.g., sorbitol syrup, cellulose derivatives or hydrogenated edible fats); emulsifying agents (e.g., lecithin or acacia); non-aqueous vehicles (e.g., almond oil, oily esters, ethyl alcohol or fractionated vegetable oils); and preservatives (e.g., methyl or propyl-p-hydroxybenzoates or sorbic acid).
  • suspending agents e.g., sorbitol syrup, cellulose derivatives or hydrogenated edible fats
  • emulsifying agents e.g., lecithin or acacia
  • non-aqueous vehicles e.g., almond oil, oily esters, ethyl alcohol or fractionated vegetable oils
  • preservatives e.g., methyl or propyl-p-hydroxybenzoates or sorbic acid.
  • the preparations may also contain buffer salts
  • Additional oral administration can be in the form of an effervescent tablet.
  • Effervescent formulations are known in the art for various active ingredients and vitamins. These effervescent formulations generally include an agent that is capable of releasing C0 2 , and an agent which induces the release of C0 2 . Suitable agents capable of releasing C0 2 which are used include alkali metal carbonates or alkali metal bicarbonates, such as sodium carbonate and sodium bicarbonate. Alkaline earth metal carbonate formulations are mainly contained in mineral preparations.
  • Suitable agents for inducing C0 2 release include edible organic acids, or their acidic salts, which are present in solid form and which can be formulated with the active ingredient and the other auxiliaries to provide granules or tablets, without premature evolution of C0 2 .
  • the active ingredients are either present in the effervescent formulation as readily soluble compounds, or they are solubilized by salt formation during the dissolution process.
  • the Thyex compositions and colostrum formulations thereof for use according to the present invention are conveniently delivered in the form of an aerosol spray presentation from pressurized packs or a nebulizer, with the use of a suitable propellant, e.g., dichlorodifluoromethane, trichlorofluorom ethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas.
  • a suitable propellant e.g., dichlorodifluoromethane, trichlorofluorom ethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas.
  • the dosage unit may be determined by providing a valve to deliver a metered amount.
  • Thyex compositions and colostrum formulations thereof of the present invention may be formulated for parenteral administration by injection by bolus injection or continuous infusion.
  • Formulations for injection may be presented in unit dosage form in ampoules or in multi-dose containers, with optionally, with an added preservative.
  • Vaccines are either those commercially available, or those prepared according to art- recognized methods, and are of various forms, including conventional forms such as aqueous dispersions, oil emulsions, liposome compositions, lyophilized forms, etc.
  • Vaccine compositions and vaccination regimens may comprise different adjuvants, emulsifiers, stabilizers, etc.
  • Vaccines are administered by different routes, including but not limited to parenteral, intramuscular, intranasal, intratracheal, subcutaneous, cutaneous, percutaneous or intracutaneous routes, and combinations thereof.
  • Vaccines may be prepared, inter alia, as aqueous solutions, syrups, elixers, or tinctures, and the liquid formulations may include suspensions and/or emulsions.
  • Thyex-4 may be lyophilized and dispensed in "00" size gelatin capsules: Oral. Approximately 40% thymic polypeptides.
  • Thyex-5 ⁇ e.g., lyophilized; approximately 80% thymic polypeptides
  • other extracts e.g., extracts containing polysaccharides such as beta 1-3 glucan.
  • the mixtures for example, can be dispensed in "00" gelatin capsules, or alternatively, for example, in size "3" capsule if not mixed with other extracts.
  • Thyex-6A e.g., sterile liquid extract
  • Thyex-6A can be used to generate aerosols ⁇ e.g., for treating pneumonia or emphysema).
  • ointments can be used when Thyex-6A is mixed with water-soluble ointment base.
  • Thyex-6BA e.g., sterile liquid buffered, and saline adjusted for injection; at least 99% pure
  • Thyex-6BA is used for veterinary and human uses.
  • Thyex compositions include, but are not limited to veterinary uses including, but not limited to treating demodectic mange and stomatitis (e.g., gingivostomatitis).
  • formulations of the present invention may include other agents known to those skilled in the art, having regard for the type of formulation in issue.
  • formulations suitable for oral administration may include flavoring agents and formulations suitable for intranasal administration may include perfumes.
  • compositions and formulations of the invention can be administered by any conventional method available for use in conjunction with pharmaceutical drugs, either as individual therapeutic agents or in a combination of therapeutic agents.
  • the dosage administered will, of course, vary depending upon known factors, such as the pharmacodynamic characteristics of the particular agent and its mode and route of administration; the age, health and weight of the recipient; the nature and extent of the symptoms; the kind of concurrent treatment; the frequency of treatment; and the effect desired.
  • injection-use formulations can be presented in unit-dose or multi-dose sealed containers, such as ampules and vials, and can be stored in a freeze-dried (lyophilized) condition requiring only the addition of the sterile liquid excipient immediately prior to use.
  • Extemporaneous injection solutions and suspensions can be prepared from sterile powders, granules, and tablets.
  • the requirements for effective pharmaceutical carriers for injectable compositions are well known to those of ordinary skill in the art. See, for example, Pharmaceutics and Pharmacy Practice, J. B. Lippincott Co., Philadelphia, Pa., Banker and Chalmers, Eds., 238-250 (1982) and ASHP Handbook on Injectable Drugs, Toissel, 4th ed., 622-630 (1986).
  • Formulations suitable for topical administration include lozenges of the compositions and optionally, an additional therapeutic and a flavor, usually sucrose and acacia or tragacanth; pastilles comprising a gas-enriched fluid and optional additional therapeutic agent in an inert base, such as gelatin and glycerin, or sucrose and acacia; and mouth washes or oral rinses comprising a gas-enriched fluid and optional additional therapeutic agent in a suitable liquid carrier; as well as creams, emulsions, gels, and the like.
  • an additional therapeutic and a flavor usually sucrose and acacia or tragacanth
  • pastilles comprising a gas-enriched fluid and optional additional therapeutic agent in an inert base, such as gelatin and glycerin, or sucrose and acacia
  • mouth washes or oral rinses comprising a gas-enriched fluid and optional additional therapeutic agent in a suitable liquid carrier
  • formulations suitable for rectal administration may be presented as suppositories by mixing with a variety of bases such as emulsifying bases or water-soluble bases.
  • bases such as emulsifying bases or water-soluble bases.
  • Suitable pharmaceutical carriers are described in Remington's Pharmaceutical Sciences, Mack Publishing Company, a standard reference text in this field.
  • the dose administered to a subject, especially an animal, particularly a human, in the context of the present invention should be sufficient to effect a therapeutic response in the animal over a reasonable time frame.
  • dosage will depend upon a variety of factors including the condition of the animal, the body weight of the animal, as well as the condition being treated.
  • a suitable dose is that which will result in a concentration of the therapeutic composition in a subject that is known to affect the desired response.
  • the size of the dose also will be determined by the route, timing and frequency of administration as well as the existence, nature, and extent of any adverse side effects that might accompany the administration of the therapeutic composition and the desired physiological effect.
  • Suitable means of administration for a particular subject will depend on the nature and severity of the disease or condition being treated or the nature of the therapy being used, as well as the nature of the therapeutic composition or additional therapeutic agent. In certain embodiments, oral or topical administration is preferred.
  • Formulations suitable for oral administration may be provided as discrete units, such as tablets, capsules, cachets, syrups, elixirs, chewing gum, "lollipop" formulations, microemulsions, solutions, suspensions, lozenges, or gel-coated ampules, each containing a predetermined amount of the active compound; as powders or granules; as solutions or suspensions in aqueous or non-aqueous liquids; or as oil-in-water or water-in-oil emulsions.
  • formulations of the present invention may include other agents known to those skilled in the art, having regard for the type of formulation in issue.
  • formulations suitable for oral administration may include flavoring agents and formulations suitable for intranasal administration may include perfumes.
  • compositions of the invention can be administered by any conventional method available for use in conjunction with pharmaceutical drugs, either as individual therapeutic agents or in a combination of therapeutic agents.
  • This example provides an exemplary process embodiment used for preparing thymus extracts, and compositions ("Thyex-1") produced in accordance therewith:
  • Thyex-1
  • Thyex-1 process comprises a process embodiment for producing Thyex-1 (step (17) relates to storage) suitable for oral delivery:
  • a volume of approximately 700 ml of 0.2% NaCl solution (in distilled water) was blended with approximately 350 g wet weight (about 400 ml wet volume) of cut-up thymus tissue in a standard size blender for at least one minute to produce a thymus homogenate;
  • step (1) Low-speed Centrifugation.
  • the "thymus homogenate" of step (1) was centrifuged at about 3,500 x G for 10 minutes at ambient temperature to produce a pellet and a supernatant fraction;
  • step (3) Crude filtration.
  • the resulting "supernatant fraction" of step (2) (after removal of any packed low density debris floating on its surface) was decanted from the centrifugation pellet and gravity filtered through one or more layers of standard cheese cloth to produce a primary filtered supernatant;
  • Steps (l)-(3) were repeated with another 350 g wet weight (about 400 ml wet volume) of prime washed, dressed, cut-up thymus glands, except that the "primary filtered supernatant" of step (3) was used in place of the 700 ml of 0.2% NaCl solution of step (1). This substitution allowed for the production of a more concentrated (relative to the "primary filtered supernatant") secondary filtered supernatant;
  • step (4) Heat denaturation.
  • the "secondary filtered supernatant" of step (4) was heated to a temperature of about 75-80°C by exposing the container thereof to a uniform heat source, such as a constant temperature water bath set at about 100°C, or a double boiler containing water at about 100°C. During said heating, the "secondary filtered supernatant” was frequently agitated or stirred until it reached about 75-80°C to produce a heat-denatured secondary filtered supernatant;
  • a uniform heat source such as a constant temperature water bath set at about 100°C, or a double boiler containing water at about 100°C.
  • step (5) Low-speed Centrifugation.
  • the "heat-denatured secondary filtered supernatant" of step (5) was centrifuged at 3,500 x g for 5 minutes at ambient temperature to produce a pellet and a heat-denatured supernatant fraction;
  • step (6) Production of a "heat-denatured filtered supernatant.
  • the "heat-denatured supernatant fraction” of step (6) was decanted from the centrifugation pellets and gravity filtered through one or more layers of standard cheese cloth to produce a filtered, heat- denatured supernatant fraction (hereinafter the "intermediate supernatant" fraction) that was still slightly warm from the heat denaturation of step (5) ;
  • step (8) Low-speed centrifugation.
  • the "salted intermediate supernatant" of step (8) was divided between two, 1 L centrifuge bottles and centrifuged at 3,500 x g for 10 minutes at ambient temperature to produce ammonium sulfate pellets, and supernatant fractions;
  • step (10) Suspension of ammonium sulfate pellet fraction.
  • the "ammonium sulfate supernatants" from step (9) were decanted from the centrifugation tubes and discarded, and excess salt solution was carefully wiped from the inside tube walls.
  • the two ammonium sulfate pellets of step (9) ⁇ i.e., corresponding to each 1-L centrifuge bottle) were then suspended and dissolved by gentle mixing with about 50 ml of 0.01 to 0.05 M phosphate buffer (about pH 7) for each pellet (alternatively, the pellets were suspended with distilled water).
  • the suspensions were allowed to stand for about 1 hour at ambient temperature with brief agitation about every 15 minutes (to facilitate complete dissolution of the pellets) to provide an ammonium sulfate fraction. Note that dissolution of any remaining ammonium sulfate pellet can be affected by the step-wise addition of small amounts of distilled water ⁇ e.g., 5 ml aliquots), followed by agitation until the pellet is completely dissolved;
  • step (10) Dialysis.
  • the "ammonium sulfate" fraction of step (10) was transferred to clean dialysis tubing ⁇ e.g., Spectrapor 3.5 kDa molecular weight cut-off size), and dialyzed with stirring ⁇ e.g., by means of a magnetically-driven stir bar in the dialysis chamber) for 3 days against an excess of distilled water at about 4°C to produce a dialyzed ammonium sulfate fraction.
  • the distilled water was changed every 12 hours.
  • Increasing hydrostatic pressure within the dialysis tubing was periodically relieved by removing some of the dialysate and transferring it to additional dialysis tubes;
  • step (12) First exclusion-membrane filtration.
  • the "dialyzed ammonium sulfate supernatant fraction" of step (12) was passed under nitrogen pressure at about 40-50 p.s.i. through a 100 kDa exclusion limit membrane filter (Amicon) at 4°C (alternatively, ambient temperature will suffice) to produce a 3.5 kDa to 100 kDa filtrate;
  • step (14) Second exclusion-membrane filtration.
  • the "3.5 kDa to 100 kDa filtrate" of step (13) was passed under nitrogen pressure at 40 to 50 p.s.i. (275.8 to 344.75 Kpa, in metric units) through a 30 kDa exclusion limit membrane filter (Amicon) to produce a 3.5 kDa to 30 kDa filtrate;
  • Thyex-1 of step (15) was filter sterilized by passage through a 0.2 micron membrane filter to produce sterile Thyex-1, suitable for oral delivery;
  • Thyex-1 produced in accordance with steps (1)-(16) of the Thyex-1 process, was typically stored frozen ⁇ e.g., -5°C to -20°C) in sterilized containers, and thawed just prior to use. According to particular aspects, the therapeutic activity of Thyex-1 was found to be stable to repeated freezing and thawing. Alternatively, Thyex-1 was lyophilized, stored at ambient temperature and reconstituted with sterile water prior to use.
  • one or more of the above steps are optional.
  • This example provides an exemplary process embodiment used for preparing thymus extracts, and compositions ("Thyex-2") produced in accordance therewith suitable for oral delivery:
  • Thyex-2 process comprises a process embodiment for producing Thyex-2 (step (9) relates to storage):
  • step (2) High-speed centrifugation.
  • the "intermediate supernatant" fraction of step (1) was cleared ⁇ i.e., to remove potential pathogens) by centrifugation at 8,500 x g for 10 minutes at ambient temperature to produce a pellet and a cleared intermediate supernatant fraction;
  • Lyophilization The "cleared intermediate supernatant” fraction of step (2) was lyophilized ⁇ i.e., freeze dried) either to complete dryness to produce a dried, cleared intermediate supernatant fraction, or until its volume was reduced by 90% to produce a lyophilized, cleared intermediate supernatant fraction;
  • step (3) Dialysis.
  • the "lyophilized, cleared intermediate supernatant,” or the alternative completely “dried” fraction (suspended in 500 ml distilled water per 13.6 kg (30 lbs.) wet weight of thymus glands processed) of step (3) was dialyzed according to step (11) of the above-identified Thyex-1 process to produce a dialyzed, lyophilized intermediate supernatant fraction;
  • step (4) High-speed centrifugation.
  • the "dialyzed, lyophilized intermediate supernatant" of step (4) was centrifuged at 8,500 x g for 10 minutes at ambient temperature to produce a pellet, and a cleared, dialyzed, lyophilized intermediate supernatant fraction;
  • step (5) Exclusion-Membrane filtration.
  • the "cleared dialyzed, lyophilized intermediate supernatant" of step (5) was passed consecutively under nitrogen pressure (40-50 p.s.i.) through 100 kDa and 30 kDa exclusion limit membrane filters (Amicon), according to steps (13) and (14) of the above-identified Thyex-1 process to produce a 3.5 kDa to 30 kDa filtrate.
  • the protein concentration of the "30 kDa filtrate” was measured, and optionally diluted (typically, to about 2 mg/0.25 ml (lesser or greater dilutions were also made as desired);
  • step (6) Adjustment of pH and ionic strength.
  • the pH and ionic strength of the "3.5 kDa to 30 kDa filtrate" or the optionally diluted "3.5 kDa to 30 kDa filtrate” of step (6) was adjusted according to step (15) of the above-identified Thyex-1 process to produce a pH- and ionic strength-adjusted 3.5 kDa to 30 kDa filtrate, Thyex-2;
  • Thyex-2 of step (7) was filter sterilized according to step (16) of the above-identified Thyex-1 process to produce sterile Thyex-2, suitable for oral delivery;
  • Thyex-2 produced in accordance with steps (l)-(8) of the Thyex-2 process was typically stored frozen ⁇ e.g., -5°C to -20°C) in sterilized containers, and thawed just prior to use. According to particular aspects, the therapeutic activity of Thyex-2 was found to be stable to repeated freezing and thawing. Alternatively, Thyex-2 was lyophilized, stored at ambient temperature and reconstituted with sterile water prior to use.
  • one or more of the above steps are optional.
  • Thyex-3 ⁇ Preparation of thymus extracts Thyex-3 :
  • Thyex-3 process comprises a process embodiment for producing Thyex-3 (step (11) relates to storage), suitable for oral delivery:
  • a volume of approximately 800 ml of 0.2% NaCl solution (in distilled water) was blended with approximately 300 g wet weight (about 340 ml wet tissue volume) of cut-up thymus tissue in a standard size blender for at least one minute to produce a thymus homogenate;
  • step (1) Low-speed Centrifugation.
  • the "thymus homogenate" of step (1) was centrifuged at about 3,500 rpm for 10 minutes at ambient temperature to produce a pellet and a supernatant fraction;
  • step (3) Crude filtration.
  • the resulting "supernatant fraction" of step (2) (after removal of any packed low density debris floating on its surface) was decanted from the centrifugation pellet and gravity filtered through one or more layers of standard cheese cloth to produce a primary filtered supernatant;
  • Steps (l)-(3) were repeated with another 175 g wet weight (200 ml wet tissue volume) of prime washed, dressed, cut-up thymus glands, except that the "primary filtered supernatant" of step (3) was used in place of the 800 ml of 0.2% NaCl solution of step (1). This substitution allowed for the production of a more concentrated (relative to the "primary filtered supernatant") secondary filtered supernatant;
  • Steps (l)-(3) were repeated with another 200 ml (wet volume) of prime washed, dressed, cut-up thymus glands, except that the "secondary filtered supernatant" from step (4) was used in place of the 800 ml of 0.2% NaCl solution of step (1). This substitution allowed for the production of a more concentrated (relative to the "primary” and “secondary filtered supernatants") tertiary filtered supernatant;
  • the "tertiary filtered supernatant" from step (5) was heated to a temperature of about 75-80°C by exposing the container thereof to a uniform heat source such as a constant-temperature water bath set at about 100°C or a double boiler containing water at about 100°C. During heating, the "tertiary filtered supernatant” was frequently agitated or stirred until it reached about 75-80°C to produce a heat-denatured tertiary filtered supernatant fraction;
  • step (6) Low-speed Centrifugation.
  • the "heat-denatured tertiary filtered supernatant" fraction of step (6) was centrifuged at 3,500 rpm for 5 minutes at ambient temperature to produce a pellet and a heat-denatured supernatant fraction;
  • step (7) Production of a "heat-denatured filtered supernatant.
  • the "heat-denatured supernatant fraction” of step (7) was decanted from the centrifugation pellets and gravity filtered through one or more layers of standard cheese cloth to produce a filtered, heat- denatured supernatant fraction that was still slightly warm from the heat denaturation of step
  • step (8) High-speed centrifugation.
  • the "filtered, heat-denatured supernatant" fraction of step (8) was centrifuged at about 8,500 x g for 5 minutes at ambient temperature to produce a pellet, and a high-speed supernatant fraction, Thyex-3;
  • step (10) Filter sterilization.
  • the "Thyex-3" fraction of step (9) was filter sterilized according to step (16) of the above-identified Thy ex- 1 process to produce sterile Thyex-3, suitable for oral delivery;
  • Thyex-3 produced in accordance with steps (1)-(10) of the Thyex-3 process was typically stored frozen ⁇ e.g., -5 to -20°C) in sterilized containers, and thawed just prior to use. According to particular aspects, the therapeutic activity of Thyex-3 was found to be stable to repeated freezing and thawing. Alternatively, Thyex-3 was lyophilized, stored at ambient temperature and reconstituted with sterile water prior to use.
  • one or more of the above steps are optional.
  • Thvex-4 (Preparation of thymus extracts Thyex-4)
  • Thvex-4 compositions
  • Thyex-4 process The following steps (l)-(l l) comprise a process embodiment for producing Thyex-4 (step (12) relates to storage), e.g., suitable for oral delivery (NOTE: the following steps (l)-(6) are referred to herein as "stage 1 steps (l)-(6)").
  • a volume of approximately 700 ml of 0.2% NaCl solution (in distilled water) was blended for at least a minute with approximately 350 g wet weight of cut-up thymus tissue in a standard size blender to produce a thymus homogenate;
  • step (1) Low-speed Centrifugation.
  • the "thymus homogenate" of step (1) was centrifuged at about 3,500 rpm for 10 minutes at ambient temperature to produce a pellet and a supernatant fraction;
  • step (3) Crude filtration.
  • the resulting "supernatant fraction" of step (2) (after removal of any packed low density debris floating on its surface) was decanted from the centrifugation pellet and gravity filtered through one or more layers of standard cheese cloth to produce a primary filtered supernatant;
  • the "primary filtered supernatant" of step (3) was heated to a temperature of about 75-80°C by exposing the container thereof to a uniform heat source, such as a constant temperature water bath set at about 100°C, or a double boiler containing water at about 100°C. During said heating, the "primary filtered supernatant” was frequently agitated or stirred until it reached about 75-80°C to produce a heat-denatured primary filtered supernatant; (5) Low-speed Centrifugation.
  • the "heat-denatured primary filtered supernatant" of step (4) was centrifuged at 3,500 x g for 5 minutes at ambient temperature to produce a pellet and a heat-denatured supernatant fraction;
  • step (5) Production of a "heat-denatured filtered supernatant.
  • the "heat-denatured supernatant fraction” of step (5) was decanted from the centrifugation pellets and gravity filtered through one or more layers of standard cheese cloth to produce a filtered, heat- denatured supernatant fraction (hereinafter the “intermediate supernatant” fraction) that was still slightly warm from the heat denaturation of step (4);
  • step (6) Dialysis.
  • the "intermediate supernatant" fraction of step (6) was dialyzed according to step (11) of the above-identified Thyex-1 process ⁇ e.g., using clean dialysis tubing ⁇ e.g., Spectrapor 3.5 kDa molecular weight cut-off size), and dialyzed with stirring ⁇ e.g., by means of a magnetically-driven stir bar in the dialysis chamber) for 3 days against an excess of distilled water at about 4°C) to produce a dialyzed, intermediate supernatant fraction;
  • step (7) Low-speed Centrifugation.
  • the "dialyzed, intermediate supernatant fraction" of step (7) was centrifuged at 3,500 rpm for 5 minutes at ambient temperature to produce a pellet and a heat-denatured supernatant fraction;
  • step (8) Production of a “heat-denatured filtered supernatant.
  • the "heat-denatured supernatant fraction” of step (8) was decanted from the centrifugation pellets and gravity filtered through one or more layers of standard cheese cloth to produce a filtered, heat- denatured supernatant fraction;
  • step (10) High-speed centrifugation.
  • the "filtered, heat-denatured supernatant" fraction of step (9) was centrifuged at about 8,500 x g for 5 minutes at ambient temperature to produce a pellet, and a high-speed supernatant fraction, Thyex-4;
  • Thyex-4" fraction of step (10) was filter sterilized according to step (16) of the above-identified Thyex-1 process to produce sterile Thyex-4, suitable for oral delivery;
  • Thyex-4 produced in accordance with steps (l)-(l l) of the Thyex-4 process was typically stored frozen ⁇ e.g., -5 to -20°C) in sterilized containers, and thawed just prior to use. According to particular aspects, the therapeutic activity of Thyex-4 is stable to repeated freezing and thawing. Alternatively, Thyex-4 was lyophilized, stored at ambient temperature and reconstituted with sterile water prior to use.
  • this example provides an exemplary process embodiment used for preparing thymus extracts, and compositions ("Thyex-5") produced in accordance therewith suitable for oral delivery:
  • Thyex-5 process comprises a process embodiment for producing Thyex-5 (step (14) relates to storage), suitable for oral delivery:
  • step (7) Low-speed centrifugation.
  • the "salted intermediate supernatant" of step (7) was divided between two, 1 L centrifuge bottles and centrifuged at 3,500 x g for 10 minutes at ambient temperature to produce ammonium sulfate pellets, and supernatant fractions;
  • step (8) Suspension of ammonium sulfate pellet fraction.
  • the "ammonium sulfate supernatants" from step (8) were decanted from the centrifugation tubes and discarded, and excess salt solution was carefully wiped from the inside tube walls.
  • the two ammonium sulfate pellets of step (8) ⁇ i.e., corresponding to each 1-L centrifuge bottle) were then suspended and dissolved by gentle mixing with about 50 ml of distilled water (or optionally with 0.01 to 0.05 M phosphate buffer (about pH 7)) for each pellet.
  • the suspensions were allowed to stand for about 1 hour at ambient temperature with brief agitation about every 15 minutes (to facilitate complete dissolution of the pellets) to provide an ammonium sulfate fraction.
  • any remaining ammonium sulfate pellet can be affected by the step-wise addition of small amounts of distilled water ⁇ e.g., 5 ml aliquots), followed by agitation until the pellet is completely dissolved;
  • step (10) Dialysis.
  • the "ammonium sulfate" fraction of step (9) was transferred to clean dialysis tubing ⁇ e.g., Spectrapor 3.5 kDa molecular weight cut-off size), and dialyzed with stirring ⁇ e.g., by means of a magnetically-driven stir bar in the dialysis chamber) for 3 days against an excess of distilled water at about 4°C to produce a dialyzed ammonium sulfate fraction.
  • the distilled water was changed every 12 hours.
  • Increasing hydrostatic pressure within the dialysis tubing was periodically relieved by removing some of the dialysate and transferring it to additional dialysis tubes;
  • step (10) High-speed centrifugation.
  • the "dialyzed ammonium sulfate fraction" of step (10) was centrifuged at 8,500 x g for 10 minutes at ambient temperature to produce a pellet and dialyzed ammonium sulfate supernatant fraction (Thyex-5);
  • step (12) Filter sterilization.
  • the "Thyex-5" of step (12) was filter sterilized by passage through a 0.2 micron membrane filter to produce sterile Thyex-5, suitable for oral delivery;
  • Thyex-5 produced in accordance with steps (1)-(13) of the Thyex-5 process, was typically stored frozen ⁇ e.g., -5°C to -20°C) in sterilized containers, and thawed just prior to use. According to particular aspects, the therapeutic activity of Thyex-5 was found to be stable to repeated freezing and thawing. Alternatively, Thyex-5 was lyophilized, stored at ambient temperature and reconstituted with sterile water prior to use.
  • one or more of the above steps are optional.
  • Thyex-6A With reference to FIGURE 4, this example provides an exemplary process embodiment used for preparing thymus extracts, and compositions ("Thyex-6A") produced in accordance therewith suitable for oral delivery, or delivery as a topical ointment or by injection or inhalation:
  • Thyex-6A process comprises a process embodiment for producing Thyex-6A (step (15) relates to storage), suitable for oral delivery:
  • step (11) First exclusion-membrane filtration.
  • the "dialyzed ammonium sulfate supernatant fraction" of step (11) was passed under nitrogen pressure at about 40-50 p.s.i. (275.8 to 344.75 Kpa, in metric units) through a 100 kDa exclusion limit membrane filter (Amicon) at 4°C (alternatively, ambient temperature will suffice) to produce a 3.5 kDa to 100 kDa filtrate;
  • step (12) Second exclusion-membrane filtration.
  • the "3.5 kDa to 100 kDa filtrate" of step (12) was passed under nitrogen pressure at 40 to 50 p.s.i. (275.8 to 344.75 Kpa, in metric units) through a 30 kDa exclusion limit membrane filter (Amicon) to produce a 3.5 kDa to 30 kDa filtrate (Thyex-6A);
  • Thyex-6A of step (13) was filter sterilized by passage through a 0.2 micron membrane filter to produce sterile Thyex-6A, suitable for oral delivery, or delivery as a topical ointment or by injection or inhalation;
  • Thyex-6A produced in accordance with steps (1)-(14) of the Thyex- 6A process, was typically stored frozen ⁇ e.g., -5°C to -20°C) in sterilized containers, and thawed just prior to use. According to particular aspects, the therapeutic activity of Thyex- 6A was found to be stable to repeated freezing and thawing. Alternatively, Thyex-6A was lyophilized, stored at ambient temperature and reconstituted with sterile water prior to use.
  • Thyex-6B process.
  • this example provides an exemplary process embodiment used for preparing thymus extracts, and compositions ("Thyex-6B") produced in accordance therewith suitable for oral delivery, or delivery as a topical ointment or by injection or inhalation:
  • Thyex-6B
  • Thyex-6B process.
  • the following steps (1)-(15) comprise a process embodiment for producing Thyex-6A (step (16) relates to storage), suitable for oral delivery, or delivery as a topical ointment or by injection or inhalation:
  • step (12) First exclusion-membrane filtration.
  • the "dialyzed ammonium sulfate supernatant fraction" of step (11) was passed under nitrogen pressure at about 40-50 p.s.i. (275.8 to 344.75 Kpa, in metric units) through a 100 kDa exclusion limit membrane filter (Amicon) at 4°C (alternatively, ambient temperature will suffice) to produce a 3.5 kDa to 100 kDa filtrate;
  • step (12) Second exclusion-membrane filtration.
  • the "3.5 kDa to 100 kDa filtrate" of step (12) was passed under nitrogen pressure at 40 to 50 p.s.i. (275.8 to 344.75 Kpa, in metric units) through a 30 kDa exclusion limit membrane filter (Amicon) to produce a 3.5 kDa to 30 kDa filtrate (Thyex-6B);
  • Thyex-6B of step (14) was filter sterilized by passage through a 0.2 micron membrane filter to produce sterile Thyex-6B, suitable for oral delivery, or delivery as a topical ointment or by injection or inhalation;
  • Thyex-6B produced in accordance with steps (1)-(15) of the Thyex- 6B process, was typically stored frozen ⁇ e.g., -5°C to -20°C) in sterilized containers, and thawed just prior to use. According to particular aspects, the therapeutic activity of Thyex- 6B was found to be stable to repeated freezing and thawing. Alternatively, Thyex-6B was lyophilized, stored at ambient temperature and reconstituted with sterile water prior to use.
  • one or more of the above steps are optional.
  • This Example 8 provides general considerations for practice of the above-identified Thyex 1-6 A and -6B process embodiments.
  • Thyex- 1 steps 1-16
  • Thyex-2 steps 1-8
  • Thyex-3 steps 1-10
  • Thyex-4 steps 1-11
  • Thyex-5 steps 1-13
  • Thyex-6A steps 1-14
  • Thyex 6B steps 1-15
  • storage ⁇ e.g., lyophilization steps of the inventive processes may be practiced with various modifications (including but not limited to those outlined below) that are within the scope of the present invention, and with alternatives or substitutions that will be recognized by those of ordinary skill in the art as being equivalent to those used herein to produce Thy ex 1-6 A and -6B.
  • Thymus glands In particular aspects, animals (e.g., steers) are taken to a packing house at about 12- 14 months. The thymus gland at this age is grayish. As an animal ages, the gland begins to become fibrous and even whitish in color. The optimum yield of final product from one kilogram (about 4.5 lb) of prime gland is 1 gram of purified Thyex (e.g., Thyex 6A or 6B). Sheep glands are generally from 6 month-old animals.
  • Thyex e.g., Thyex 6A or 6B
  • Freshly harvested thymus glands from porcine, ovine, or bovine sources should optimally be frozen within 24 hours of harvest and stored frozen, preferably for at least 72 hours. Freezing of the thymus glands renders the cells more susceptible to disruption in isotonic or more preferably hypotonic ⁇ e.g., 0.2% to 0.3% salt, such as NaCl) salt solution during homogenization. Variations in the freezing temperature and duration are within the scope of the present invention.
  • the thymus glands are preferably frozen at least once ⁇ e.g., -5 to -20°C) for production of optimal extracts.
  • thawed glands are preferably first washed and extraneous materials, such as fatty tissues, lymph nodes, and connective tissues are preferably excised and discarded.
  • the tissues are preferably minced into approximately 1" squares before subjected to grinding (e.g., in a food processor, meat grinder, blender, or equivalent or suitable device).
  • the ground glands are homogenized in a blender at a proportion of about 350 gm wet weight with 700 ml of 0.2% saline for at least a minute.
  • the supernatant solution (upper and lower ppt discarded) is heat denatured by raising the solution temperature in a double boiler with constant stirring to a temperature in the range of about 75°C to 80°C (preferably 75°C).
  • the supernatant solution is collected and precipitate (ppt) discarded.
  • the glands for all Thyex processes 1, 2, 3, 4, 5, 6A, and 6B are processed through this phase in identical or very similar fashion.
  • step (4) of the Thyex- 1 process embodiment allows for more concentrated filtered supernatants (relative to the corresponding "primary”-filtered supernatants), thus reducing the amount of ammonium sulfate required (Thyex- 1 process embodiment), or the lyophilization time required (Thyex-2 process embodiment) to process a given amount of thymus tissue.
  • variations in the final protein concentrations (e.g., in the range of 1 to 7 mg/ml) of the various primary-, secondary- and tertiary-filtered supernatants reflect the average age of the animals from which thymus tissue is obtained.
  • the protein concentration of the tertiary-filtered supernatant is about 4 mg/ml.
  • a heat-denaturation step is integral to all of the above-described Thyex process embodiments, and facilitates precipitation and subsequent removal of relatively large, heat- labile proteins that have no utility in the claimed compositions or methods (see below).
  • Variation in the volume of filtered supernatant fraction treated, in the final temperature of the heat-denaturation step (within the range of about 75°C to about 80°C), in the temperature of the uniform heat source (within the range of about 80°C to about 100°C, preferably about 100°C) and in the time period over which heating of the filtered supernatant fractions from initial to said final temperature takes place are within the scope of the present invention.
  • the supernatant is heated to the final temperature at a rate that is as rapid as possible whereby said rate, in combination with stirring, generally minimizes the occurrence of local supernatant temperatures (e.g., supernatant temperatures near the heat-transferring wall of the supernatant container) that exceed the desired final temperature.
  • local supernatant temperatures e.g., supernatant temperatures near the heat-transferring wall of the supernatant container
  • duration and frequency of stirring during said heating are within the scope of the present invention, and depend on the temperature of the constant- temperature heat source and the volume of supernatant being heated. Generally, both the duration and frequency of stirring increase with increasing supernatant volume or heat- source temperature. Constant stirring is also effective, and preferable when heating relatively large supernatant volumes.
  • Step (8) of the above-described Thyex- 1 process embodiment, and step (7) of the above-described Thyex-5, -6A and -6B process embodiment involves protein concentration/fractionation by ammonium sulfate precipitation of the "intermediate supernatant" fraction.
  • solid ammonium sulfate is added to attain high salt concentrations (e.g., in excess of about 0.7 gm/ml) with minimal dilution.
  • this concentration/fractionation step is achieved by adding saturated ammonium sulfate solution.
  • this embodiment results in relatively lower final salt concentrations (e.g., of about 0.5 gm/ml or greater), and is thus less efficient in precipitating (and thereby recovering) desirable low molecular weight proteins. Nonetheless, according to particular aspects, the resulting Thyex compositions have activity in the claimed methods, albeit to a lesser degree. Moreover, the present invention also encompasses the use of combinations of saturated or sub-saturated ammonium sulfate solutions with solid ammonium sulfate.
  • a dialysis steps of the above-described Thyex process embodiments allow any molecules of molecular weight less that about 3.5 kDa to pass through.
  • Variation in the precise exclusion limit of the dialysis membrane is within the scope of the present invention.
  • any dialysis membrane is acceptable provided that its exclusion limit (porosity) enables the retention of molecules having molecular weights of about 5 kDa or larger.
  • any such filtration membrane is acceptable provided that its exclusion limit (porosity) does not result in exclusion (i.e., removal from the final Thyex composition) of molecules having molecular weights equal to or smaller than about 15 kDa.
  • exclusion membranes that exclude molecules of about 20, 30 or 40 kDa or larger are useful in the practice of the present invention, but result in final Thyex compositions that are less active per mg of final protein, compared to those compositions prepared using an exclusion membrane the excludes proteins larger than about 15 kDa.
  • dialysis and filtration membranes are chosen such that the resulting Thyex compositions comprise proteins in the molecular weight range of about 5 to 14 kDa.
  • the process embodiments may further comprise fractionation, based on molecular weight, to obtain a final protein fraction having proteins of about 3.5 to about 30 kDa.
  • membrane filters e.g., cellulose acetate membranes; Millipore
  • Some commercially-available membrane filters are self-contained and provided as pre-sterilized, disposable units.
  • Other membranes are mounted in reusable membrane holders, and heat sterilized in an autoclave prior to use.
  • the final Thyex 1-6 A and -6B compositions are standardized at a protein concentration about 2 mg/ml, based on optical density at 260 and 280 nm. Preferred dosages are discussed herein above under “Dose Determinations.”
  • Thyex 6A and Thyex 6B process embodiments are designed to provide therapeutic compositions suitable for delivery as a topical ointment or by injection or inhalation, and include ammonium sulfate precipitation/fractionation steps.
  • Thyex-5 is prepared from a similar process but is somewhat less refined than Thyex-6A or Thyex-6B, and is designed to be preferably mixed in appropriate ratios with extracted lyophilized herbal sources and administered orally in, for example, filled gelatin capsules.
  • the Thyex-4 process embodiment lacks ammonium sulfate precipitation step but provides for a sufficiently-concentrated composition after lyophilization.
  • the resulting Thyex-4 composition is less refined in relative to those of Thyex-5 (and Thyex-6A and -6B) but is nonetheless suitably concentrated and formulated for efficacious oral delivery in both animals and humans.
  • the thymus extract employed (Kyosenex®) was available in oral use form and injectable form. Since the cat was hard to medicate, we opted for ubcutaneous injection.
  • One vial of lyophylized thymic extract was hydrated with 2.0 cc of diluent (standard saline; final protein concentration about 2 mg/ml).
  • the cat was given a five day cycling dosing pattern that consisted of giving 0.2 cc of this mixture by subcutaneous injection daily for three days and skipping two days before repeating the cycle again. During this time the cat also received acupuncture weekly for three total treatments. After a month, the dose was reduced to 0.1 cc daily for three days, off for two days and repeated in five day cycles.
  • Figure 5 shows day one of therapy. Note multiple extractions and excellent healing from prior dental care. At this time gingivostomatitis-related inflammation is very visible, and the patient objected strongly to opening her mouth.
  • Figure 6 shows thirty days of therapy (6 x 5 day cycles). Only some residual inflammation is still present, and the mouth is far less red and far more comfortable. Cat was eating well and playing at this exam. She did not object to opening her mouth.
  • Figure 7 shows sixty days of therapy (12 x 5 day cycles). Some residual inflammation is still present, particularly in the incisor and canine tooth regions, but the pharyngeal area is nearly normal.
  • treatment of stomatitis e.g., gingivostomatitis
  • thymus extracts e.g., Thyex-l-Thyex 6 A and Thyex-6B, Kyosenex®
  • additional agent e.g., thymus extract in combination with at least one of antibiotics, medicated mouth rinses, salt water, hydrogen peroxide, xylocaine, antiviral agents, aciclovir, fluid intake, good oral hygiene, gentle debridement of the mouth, etc.
  • the thymus extract was used in an injectable form.
  • similar therapeutic efficacy was achieved in multiple additional animal subjects using oral administration of the thymus extract (in an orally deliverable form as described herein).
  • administration by injection, oral administration, inhalation, topically, or rectal administration is efficacious in treating stomatitis (e.g., gingivostomatitis) with thymus extracts (e.g., Thyex-l-Thyex 6A and Thyex-6B, Kyosenex®), including in combination with at least one additional agent.
  • Figure 8 shows arrival of demodectic mange patient at the animal control unit prior to Kyosenex® treatment.
  • Treatment had been initiated a week earlier with oral ivermectin (0.5 cc PO SID).
  • ivermectin is a broad-spectrum antiparasitic agent, traditionally against worms. More recent evidence supports its off-label use against arthropods.
  • the main concern is neurotoxicity, which in most mammalian species may manifest as central nervous system depression, and consequent ataxia, as might be expected from potentiation of inhibitory GABA-ergic synapses. Dogs with defects in the P-glycoprotein gene can be severely poisoned by ivermectin. Moreover, this patient was not responding to ivermectin, and her condition was worsening.
  • Kyosenex® was initiated at 0.5 cc subcutaneous injections three times a week.
  • One vial of lyophylized thymic extract was hydrated with 2.0 cc of diluent (standard saline; final protein concentration about 2 mg/ml).
  • Figure 8 shows arrival of demodectic mange patient at the animal control unit prior to Kyosenex® treatment.
  • Figure 9 shows initiation of Kyosenex® treatment; Day 1 (day 7 of ivermectin treatment). Facial and truncal skin exhibited excessive skin folds and hyperemic and erythemic pustules. Examination after one week of treatment revealed a dramatic improvement, with beginning regrowth of the hair coat and significant decrease in skin pustules, erythema and edema (Figure 10).
  • Figure 10 shows day 8, beginning of second week of Kyosenex® treatment (3 injections total).
  • Figure 13 shows day 28, after discontinuation of Kyosenex® treatment (12 injections total). Note, in comparison with Figures 10 and 11, the regrowth of haircoat along the trunk. At that time (day 28), the hair coat looked normal. The patient appeared happy, friendly, exuberant, and interacted well with other dogs and people. The rapidity and extent of response when Kyosenex® was added to her initial treatment protocol was much more significant than expected based on experience with the use of ivermectin only. Shortly after her treatment protocol was completed, the patient was adopted to a permanent home (Figure 14). At one- year post treatment, there has been no recurrence of the condition. Figure 14 shows patient in her permanent home, 2-months post-treatment with Kyosenex®.
  • treatment of demodectic mange with thymus extracts e.g., Thyex-l-Thyex 6 A and Thyex-6B, Kyosenex®
  • thymus extracts e.g., Thyex-l-Thyex 6 A and Thyex-6B, Kyosenex®
  • additional agent e.g., thymus extract in combination with at least one of rotenone, Goodwinol (a rotenone-based insecticide ointment), antibiotics, medicated shampoos, Amitraz (a parasiticidal dip), avermectins, ivermectin, doramectin, milbemycin, and sulfurated lime, etc.
  • the thymus extract was used in an injectable form (subcutaneous injection).
  • similar therapeutic efficacy is achieved using other administration routes, including oral administration of the thymus extract (in an orally deliverable form as described herein).
  • administration by injection, oral administration, inhalation, topically, or rectal administration is efficacious in treating demodectic mange with thymus extracts (e.g., Thyex-l-Thyex 6A and Thyex-6B, Kyosenex®), including in combination with at least one additional agent.
  • thymus extracts e.g., Thyex-l-Thyex 6A and Thyex-6B, Kyosenex®
  • administering the inventive Thyex compositions along with at least one other antibacterial, antifungal, antiviral agent and/or along with a polysaccharide extract to stimulate macrophage achieves a most treatment response.
  • administration of Thyex extract alone is sufficient.
  • treatment of stomatitis e.g., gingivostomatitis
  • thymus extracts e.g., Thyex- 1 -Thyex 6 A and Thyex-6B, Kyosenex®
  • additional agent e.g., thymus extract in combination with at least one of antibiotics, medicated mouth rinses, salt water, hydrogen peroxide, xylocaine, antiviral agents, aciclovir, fluid intake, good oral hygiene, gentle debridement of the mouth, colostrum, etc.
  • antibiotics e.g., medicated mouth rinses, salt water, hydrogen peroxide, xylocaine, antiviral agents, aciclovir, fluid intake, good oral hygiene, gentle debridement of the mouth, colostrum, etc.
  • treatment of demodectic mange with thymus extracts e.g., Thyex- 1 -Thyex 6 A and Thyex-6B, Kyosenex®
  • thymus extracts e.g., Thyex- 1 -Thyex 6 A and Thyex-6B, Kyosenex®
  • additional agent e.g., thymus extract in combination with at least one of rotenone, Goodwinol (a rotenone-based insecticide ointment), antibiotics, medicated shampoos, Amitraz (a parasiticidal dip), avermectins, ivermectin, doramectin, milbemycin, and sulfurated lime, colostrum, etc.
  • Beta glucans are a preferred agent in combination with the inventive Thyex compositions, and act synergistically in combating demodectic mange and stomatitis (e.g., gingivostomatitis) and other related conditions.
  • beta glucan There are three forms of beta glucan based on the linkages of the complex sugars, and these are recognized as beta- 1,3 or 1,4, and 1,6 glucan. Most are in the form of 1,3 and 1,4, or 1,3 and 1,6, but the 1,3 form, which is most abundant in the fruiting bodies of certain mushrooms (e.g., Sparassis crupa or Cauliflower mushroom; or Lentinula edodes or shitake, etc.). Typically, marketing strategies relating to marketing glucan (typically from the cell wall of the common yeast) emphasize “enhanced the immune system,” “increases antibody production,” and "fight cancer.”
  • a preferred polysaccharide comprises one or more of the beta glucans, including three types based on the linkages: 1-3, 1-4, and 1-6).
  • a number of commercial beta glucan products are available with most being derived from the common yeast.
  • the preferred sources are mushrooms; with Shitake being most common because of its ready availability/source, and cauliflower mushroom (Sparassis crupa), which is preferred as it contains beta 1-3 glucans, but unfortunately has limited availability. Additionally, the shitake mushroom, which is most widely available, is reported to contain the 1-3 glucan and chitin.
  • an oral route of administration is favorable, possibly because the intestinal walls are sites containing large amounts of lymph nodes and thus T cells.
  • Colostrum formulations Additional combinations of the thymus extracts with colostrum (e.g., bovine colostrum) are disclosed herein (see working Example 13), and provide for reducing itching as described herein.
  • colostrum e.g., bovine colostrum
  • bovine colostrum is used to formulate the thymus extract for administration, and in certain embodiments, first milking bovine colostrum is used (e.g., "First Milking Bovine ColostrumTM” from Immuno-Dynamics, Fennimore, WI; sold as "ID- 1" optionally with methyl paraben and/or propyl paraben as preservative).
  • first milking bovine colostrum is used (e.g., "First Milking Bovine ColostrumTM" from Immuno-Dynamics, Fennimore, WI; sold as "ID- 1" optionally with methyl paraben and/or propyl paraben as preservative).
  • Kyosenex® PRIME Canine is formulated (formulated for Applicant by Immuno-Dynamic & URL Laboratories, Fennimore, WI USA, using ID-1 serum ("Bovine IgG Colostrum Serum”)) using 66.6 ug of Kyosenex/ml colostrum and administered in 7/10 ml (0.7 ml) per spray or dropwise, twice daily, 1 spray for each 15 lbs until resolution of condition, and then twice a week thereafter for maintenance.
  • ID-1 serum Bovine IgG Colostrum Serum
  • Kyosenex® PRIME Feline is formulated (formulated for Applicant by Immuno-Dynamic & URL Laboratories, Fennimore, WI USA, using ID-1 serum ("Bovine IgG Colostrum Serum”)) using 66.6 ug of Kyosenex/ml colostrum and administered in 7/10 ml (0.7 ml) per spray or dropwise), twice daily, 1 spray for each 15 lbs until resolution of condition, and then twice a week thereafter for maintenance.
  • ID-1 serum Bovine IgG Colostrum Serum
  • Kyosenex® PRIME Avian is formulated (formulated for Applicant by Immuno-Dynamic & URL Laboratories, Fennimore, WI USA, using ID-1 serum ("Bovine IgG Colostrum Serum”)) using 66.6 ug of Kyosenex®/ml colostrum and administered in 7/10 ml (0.7 ml) per spray or dropwise, twice daily, 1 spray for each 15 lbs until resolution of condition, and then twice a week thereafter for maintenance.
  • preferred aspects comprise treatment of demodectic mange and stomatitis (e.g., gingivostomatitis) using Thyex compositions in combination with colostrum and/or other fungal and/or herbal preparations, etc., including the following:
  • Paresis crepe (aka cauliflower mushroom or hanabaritake) preparations, comprising beta 1-3 glucan, can be used to stimulate macrophage in combination with the inventive Thyex compositions.
  • Lentinula edodes (shitake; e.g., alkaline digest according to the procedure reported by Ohno et al. (Biol. Phar. Bull. 23 866-872, 2000), comprises beta 1-3 glucan and chitin, and can be used for treating age-related illness in combination with the inventive Thyex compositions.
  • Astralagas membranaceus (Scutellaria baicalensis, Houttuynia cordata; hot water extract of ground herbs and secondary extraction by alkaline digest as above), stimulate macrophages, and can be used for treating age-related illness in combination with the inventive Thyex compositions.
  • Houttuynia cordata extracts from leaves (see Applicant's U.S. Patent No. 8,609,824), incorporated by reference herein in its entirety for teaching on Houttuynia cordata extracts) can be used for treating age-related illness in combination with the inventive Thyex compositions.
  • Houttuynia cordata has substantial utility to treat nausea in both humans and animals caused by illnesses, infections, or other treatments, and in particular embodiments is used in combination with one or more of the inventive Thyex compositions, plus or minus standard additional drugs.
  • Treatment in combination with at least one of traumeel, homotox, SBGA (blue green algae), placenta, wobenzyme, spascupreel, IMM formula, yunnan paiyo, vitamin E, omega- 3 fatty acids, semongrass oil, and cedar oil is also provided.
  • Thyex compositions in combination with colostrum was effective in reducing allergic itching
  • a thymic extract (Kyosenex®) was formulated with colostrum and used in management of a severe case of canine itching relating to allergy. The patient responded dramatically and favorably.
  • Subject A male dog showing allergic symptoms, presented with pruritic, red skin accompanied by sever itching and inflammation.
  • Kyosenex® was formulated with colostrum. Specifically, for this example, Kyosenex® PRIME Canine was formulated (formulated for Applicant by Immuno-Dynamic & URL Laboratories, Fennimore, WI USA, using ID-1 serum ("Bovine IgG Colostrum Serum”)) using 66.6 ug of Kyosenex®/ml colostrum and administered in 7/10 ml (0.7 ml) per spray or dropwise, twice daily, 1 spray for each 15 lbs subject weight until resolution of condition, and then twice a week thereafter for maintenance.
  • ID-1 serum Bovine IgG Colostrum Serum
  • the treatment protocol continued for a total of 5 days, with improvement each day.
  • the rapidity and extent of response when the Kyosenex® / colostrum formulations was administered was dramatic, and much more significant than expected based on experience with the use of other agents, including compared with experience based on use of homeopathic remedies and/or Kyosenex® alone.
  • thymus extracts e.g., Thyex-l-Thyex 6A and Thyex-6B, Kyosenex®
  • colostrum a colostrum
  • the thymus extract colostrum formulation was administered in an aerosol (oral spray) form.
  • similar therapeutic efficacy is achieved using various administration routes, including administration orally, injection, inhalation, topically, or rectal administration is efficacious in treating itching with thymus extracts (e.g., Thyex-l-Thyex 6A and Thyex-6B, Kyosenex®) formulated with colostrum, including in combination with at least one additional agent (e.g., herbs or homeopathics).
  • thymus extracts e.g., Thyex-l-Thyex 6A and Thyex-6B, Kyosenex®
  • additional agent e.g., herbs or homeopathics
  • the dosage administered will vary, depending upon the size, needs and responsiveness of the subject.
  • 66.6 ug of Kyosenex®/ml colostrum is typical, less Kyosenex®/ml colostrum can be used.
  • 33.3 ug Kyosenex®/ml colostrum can be used, or 15 ug Kyosenex®/ml colostrum can be used.
  • the amount of Kyosenex®/ml colostrum can vary between 5 to 100 ug of Kyosenex®/ml colostrum, more preferably between 10 to 80 ug of Kyosenex®/ml colostrum, even more preferably between 20 to 80 ug of Kyosenex®/ml colostrum, and most preferably between 30 to 70 ug of Kyosenex®/ml colostrum is used.
  • preservatives e.g., methyl paraben and/or propyl paraben
  • preservatives e.g., methyl paraben and/or propyl paraben
  • colostrum may serve, inter alia, as a mucosal coating that increases the tissue exposure of the Kyosenex® polypeptides, and thereby increasing absorption
  • the numbers expressing quantities of ingredients, properties such as molecular weight, reaction conditions, and so forth, used to describe and claim certain embodiments of the application are to be understood as being modified in some instances by the term "about.” Accordingly, in some embodiments, the numerical parameters set forth in the written description and attached claims are approximations that can vary depending upon the desired properties sought to be obtained by a particular embodiment. In some embodiments, the numerical parameters should be construed in light of the number of reported significant digits and by applying ordinary rounding techniques. Notwithstanding that the numerical ranges and parameters setting forth the broad scope of some embodiments of the application are approximations, the numerical values set forth in the specific examples are reported as precisely as practicable.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Animal Behavior & Ethology (AREA)
  • Epidemiology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Medicinal Chemistry (AREA)
  • Immunology (AREA)
  • Zoology (AREA)
  • Virology (AREA)
  • Biomedical Technology (AREA)
  • Developmental Biology & Embryology (AREA)
  • Engineering & Computer Science (AREA)
  • Cell Biology (AREA)
  • Biotechnology (AREA)
  • Molecular Biology (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

L'invention concerne des procédés pour traiter le prurit provoqué par une allergie, notamment le prurit associé à une inflammation due à un parasite (par exemple, la démodicose, la stomatite, la dermatophytose, etc.), comprenant l'administration, à un sujet mammifère en ayant besoin, d'une quantité thérapeutiquement efficace d'une composition d'extrait de thymus fractionnée, traitée thermiquement (par exemple, des compositions de Thyex-1-6A et -6B comprenant des protéines ou des polypeptides, par ex. ayant des masses moléculaires comprises dans la plage de 3,5 kDa à 30 kDa), combinée ou formulée avec du colostrum, afin de réduire le prurit chez le sujet. L'invention concerne également des polythérapies ou des traitements auxiliaires comprenant l'administration d'une composition d'extrait de thymus fractionnée, traitée thermiquement, combinée ou formulée avec du colostrum et contenant éventuellement au moins un agent anti-parasitaire, antibactérien, antifongique, antiviral supplémentaire, ou un agent homéopathique.
PCT/US2017/029961 2016-05-02 2017-04-27 Compositions et méthodes de traitement du prurit provoqué par une allergie Ceased WO2017192360A1 (fr)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US201662330778P 2016-05-02 2016-05-02
US62/330,778 2016-05-02
US15/206,145 2016-07-08
US15/206,145 US10842819B2 (en) 2014-01-09 2016-07-08 Compositions and methods for treating itching, gingivostomatitis, and demodectic mange

Publications (1)

Publication Number Publication Date
WO2017192360A1 true WO2017192360A1 (fr) 2017-11-09

Family

ID=60203219

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2017/029961 Ceased WO2017192360A1 (fr) 2016-05-02 2017-04-27 Compositions et méthodes de traitement du prurit provoqué par une allergie

Country Status (1)

Country Link
WO (1) WO2017192360A1 (fr)

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5750149A (en) * 1993-01-26 1998-05-12 Horse Vitality Ltd Pharmaceutical and dermocosmetic compositions containing equine colostrum
US20130274177A1 (en) * 2008-12-27 2013-10-17 Pawan Saharan Mammalian colostrum derived nanopeptides for broadspectrum viral and recurrent infections with a method of isolation thereof
WO2015105905A1 (fr) * 2014-01-09 2015-07-16 Cmi Research Management, Llc Traitement de la gingivo-stomatite et de la gale démodectique

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5750149A (en) * 1993-01-26 1998-05-12 Horse Vitality Ltd Pharmaceutical and dermocosmetic compositions containing equine colostrum
US20130274177A1 (en) * 2008-12-27 2013-10-17 Pawan Saharan Mammalian colostrum derived nanopeptides for broadspectrum viral and recurrent infections with a method of isolation thereof
WO2015105905A1 (fr) * 2014-01-09 2015-07-16 Cmi Research Management, Llc Traitement de la gingivo-stomatite et de la gale démodectique

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
CONTACT STOMATITIS CLINICAL PRESENTATION (MEDSCALE, 7 October 2015 (2015-10-07), Retrieved from the Internet <URL:http://emedicine.medscape.com/article/1076589-clinical> [retrieved on 20170629] *
SYMPTOMS OF RINGWORM INFECTIONS (CENTERS FOR DISEASE CONTROL AND PREVENTION, 6 December 2015 (2015-12-06), Retrieved from the Internet <URL:https://www.cdc.gov/fungal/diseases/ringworm/symptoms.html> [retrieved on 20170629] *

Similar Documents

Publication Publication Date Title
US9006401B2 (en) Methods of treatment using thymus-derived compositions
AU2007255983A1 (en) Cationic milk proteins for treating mastitis
JP2002501526A (ja) 乳汁における免疫グロブリンａの生成方法
EP3082841B1 (fr) Méthodes de traitement d&#39;un animal
RU2619862C1 (ru) Способ лечения субклинического мастита у лактирующих коров
CN106075436B (zh) 一种不含抗生素的抗幽门螺杆菌口服制剂
EP0484148A1 (fr) Procédé de préparation d&#39;un médicament pour à la fois, le traitement et la prévention d&#39;ulcères peptiques gastriques et de la gastrite, et des médicaments ainsi formulés
WO2015105905A1 (fr) Traitement de la gingivo-stomatite et de la gale démodectique
ES2856673T3 (es) Composiciones IgA secretoras, métodos de fabricación y métodos de uso de las mismas
US10842819B2 (en) Compositions and methods for treating itching, gingivostomatitis, and demodectic mange
RU2441660C1 (ru) Способ лечения субклинического мастита у лактирующих коров
US9249188B2 (en) Mammalian colostrum derived nanopeptides for broadspectrum viral and recurrent infections with a method of isolation thereof
WO2017192360A1 (fr) Compositions et méthodes de traitement du prurit provoqué par une allergie
RU2320167C1 (ru) Способ повышения адаптационных возможностей овец
RU2262350C2 (ru) Вакцина для иммунопрофилактики и иммунотерапии заболеваний человека и животных, вызванных патогенными и условно-патогенными грамотрицательными микроорганизмами кишечной группы и их экзотоксинами, и способ ее получения (варианты), иммуноглобулиновый препарат (варианты) и способ его получения, иммунобиологический препарат, поликомпонентная вакцина
EP4415726A1 (fr) Compositions thérapeutiques et préventives
RU2404770C1 (ru) Способ получения комплексного иммунотропного препарата для животных
US20070212367A1 (en) Novel immunologically active peptide fragments of a proline-rich polypeptide isolated from colostral mammalian fluids for treatment of viral and non-viral diseases or diseased conditions
TWI789123B (zh) 使用生長因子來治療關節炎
RU2620548C1 (ru) Способ профилактики респираторных болезней телят
CN101797384B (zh) 三联复合免疫增强剂及其制备方法
TW202321280A (zh) 使用生長因子來治療關節炎
SU1651905A1 (ru) Способ лечени дикроцелиоза овец
AU2006332821A1 (en) Novel immunologically active peptide fragments of a Proline-Rich Polypeptide isolated from colostral mammalian fluids for treatment of viral and non-viral diseases or diseased conditions

Legal Events

Date Code Title Description
NENP Non-entry into the national phase

Ref country code: DE

121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 17793031

Country of ref document: EP

Kind code of ref document: A1

32PN Ep: public notification in the ep bulletin as address of the adressee cannot be established

Free format text: NOTING OF LOSS OF RIGHTS PURSUANT TO RULE 112(1) EPC (EPO FORM 1205A DATED 17/04/2019)

122 Ep: pct application non-entry in european phase

Ref document number: 17793031

Country of ref document: EP

Kind code of ref document: A1