IE20150193A1

IE20150193A1 - Kinetin (N6-Furfuryadenine) for preventing and treating mucositis and proctitis and erythemia

Info

Publication number: IE20150193A1
Application number: IE20150193A
Authority: IE
Inventors: Shanahan-Prendergast Elizabeth
Original assignee: Leo Shanahan-Prendergast; Shanahan-Prendergast Elizabeth
Priority date: 2015-07-01
Filing date: 2015-07-01
Publication date: 2017-01-11

Abstract

The present invention provides a method for the prophylaxis and treatment of mucositis, proctitis and Erythemia resultant from radiation or chemotherapy for cancer. The present invention discovered and valadidated that Kinetin (N6-Furfuryadenine) a known dietary supplement already with medical utility in familial Dysautonomia can prevent and/treat mucositis, proctitis and erythemia in cancer patients.

Description

KINETIN (N6~Furfuryadenine) FOR PREVENTING AND TREATING MUCOSITIS AND PROCTITIS AND ERYTHEMIA Field of the Invention The present invention provides compositions and methods for the prophylaxis and/or treatment of mucositis, proctitis and Erythemia More specifically, the present invention provides composition of oral doses of Kinetin (N6-Furfuryadenine) and oral rinse suspension dosage forms and their use in the prevention and treatment of mucositis, proctitis and erythemia in subjects undergoing radiotherapy and or chemotherapy.

Background to the Invention Mucositis is an inflammatory condition of the mucous membranes or mucosa lining the digestive tract. The condition is caused by a breakdown of the mucosa, which results in the formation of ulcerative lesions. These lesions can be extremely painful and can occur at sites in the alimentary tract from the oral cavity to the anus, including the oesophagus, stomach, small intestine, colon and rectum.

Mucositis is a common side effect of chemotherapy or radiotherapy. The mucosa of the mouth and digestive tract are sensitive to both chemotherapy and radiotherapy. The chemotherapeutic agents used to treat cancerous conditions adversely affect normal cells, in particular those which have high turnover rates, such as the cells of the ora! epithelial tissues. These radiation therapy treatments cause cell death, which results in the mucosal lining becoming thin, sloughed off and then red, inflamed and ulcerated Patients undergoing chemotherapy usually become symptomatic within four to five days of commencing treatment.

Mucositis associated with radiotherapy generally presents within 14 days of treatment, with the symptoms persisting for 6 to 8 weeks.

The pathophysiology of mucositis can be divided into five stages including an initiation stage, a message generation stage, a signalling and amplification stage, an -2ulceration stage and a healing stage. The different stages are caused by different cytokines. The initiation stage follows chemotherapy or radiotherapy, which results in the production of free radicals which cause DNA damage. In turn, transcription factors, such as NF-kB, are produced that up regulate inflammatory cytokine production. This inflammation, which is mediated by cytokines such as IL-1 and TNF-alpha, causes the ulceration stage.

The main clinical manifestations of mucositis include esophagitis (inflammation of the oesophagus), dysphagia (difficulty in swallowing), odynophagia (painful swallowing), substernal chest pain (in radiation induced mucositis) and retrosternal chest pain (caused by chemotherapy).

There are no effective treatment or preventative for mucositis. Current treatments are generally palliative and include maintaining a high level of oral hygiene, the use of topical analgesics, such as lidocaine, and mouthwashes, such as chlorohexidine gluconate. Further therapies include the use of agents which reduce the mucosal absorption of chemotherapy drugs, for example cryotherapy or allopurinol. Other treatments, such as glutamine or beta- carotene, reduce changes in epithelial proliferation. Further treatments include laser therapy and antibiotics, as well as the use of cytokine-based therapies, such as palifermin (brand name Kepivance, Amgen), which is a human keratinocyte growth factor (KGF), and other modulators of inflammation.

None of the currently used therapeutic approaches has proved entirely effective in the prophylaxis or treatment of mucositis. There is therefore a substantial unmet clinical need for therapies which can be used for the effective prophylaxis and treatment of mucositis. Such therapies will be particularly beneficial to patients presenting with cancerous conditions who will undergo, or who are undergoing, cancer therapy such as chemotherapy and/or radiotherapy. -3The inventor has surprisingly identified a novel compound Kinetin (N6Furfuryadenine) which provides a methods for the prevention or treatment of mucositis in subjects who are undergoing radiation therapy. This compound is available as a dietary Supplement and has shown success in the treatment of patients with Familial Dysautonomia in New York University School of Medicine when administered at doses up to 30mg/kg for up to three years with limited toxicity. The inventor has also identified compositions and formulations of Kinetin having utility in the prophylaxis and treatment of proctitis and erythemia in subjects undergoing cancer treatment by radiotherapy.

Summary of the invention According to a first aspect of the present invention there is provided a method for the prophylaxis and/or treatment of mucositis, proctitis and erythemia the method comprising the steps of -providing a therapeutically effective amount of a composition comprising Kinetin (N6-Furfuryadenine) or an analogue, derivative, metabolite, prodrug, solvate or pharmaceutically acceptable salt thereof; and - administering the composition to a subject.

According to a second aspect of the present invention there is provided the use of a composition comprising at least one compound Kinetin (Νό-Furfuryadenine) or an analogue, derivative, metabolite, prodrug, solvate or pharmaceutically acceptable salt thereof in the preparation of a medicament for the treatment and/or prophylaxis of mucositis,proctitis and erythemia According to a third aspect of the present invention there is provided a composition for use in the prevention and/or treatment of mucositis,proctitis and erythemia the composition comprising Kinetin (N6-Furfuryadenine) or an analogue, derivative, metabolite, prodrug, solvate or pharmaceutically acceptable salt thereof According to a forth aspect of the present invention there is provided a pharmaceutical composition comprising at least one compound selected from Kinetin (N6-Furfuryadenine) or an analogue, derivative, metabolite, prodrug, solvate or -4pharmaceutically acceptable salt thereof along with a pharmaceutically acceptable carrier.

In certain embodiments, the compound of the invention is co-administered along with, 10 or formulated with, carboxymethyl cellulose (CMC).

In certain embodiments, the Klnetin compound of this patent is administered with, or formulated with, a chemotherapeutic agent which may be selected from the group consisting of, but not limited to, cisplatin, dexamethasone and 5-fluorouracil.

Detailed Description of the Invention Without wishing to be bound by theory, the invention is based, in part, on the inventor's unexpected finding that treatment of a subject with the compound Kinetin of the invention can prevent thinning and ulceration of the mucosa of the digestive (GI) tract of a subject.

The term mucositis as used herein is intended to comprise alimentary mucositis. In certain embodiments, the alimentary mucositis comprises oral mucositis and/or enteritis (inflammation of the intestines, in particular the small intestine). In certain embodiments, the alimentary mucositis comprises esophagitis (inflammation of the esophagus), oropharyngeal mucositis, stomatitis (inflammation of the stomach) and/or proctitis (inflammation of the rectum) In certain embodiments, the methods and uses of the present invention comprise administering a therapeutically effective amount of Kinetin to at least one area of the digestive tract of a subject with mucositis or at risk of developing mucositis. In certain embodiments, this compound may be administered to more than one area of the alimentary canal.

In certain embodiments, the compositions, methods and uses extend to preventing mucositis ,proctitis and erythemia in a subject who is to undergo radiation therapy and/or chemotherapy. In certain embodiments, the subject may be administered the compound Kinetin prior to conditioning myeloablative radiation therapy and/or chemotherapy in preparation for autologous or allogenic haematopoietic stem cell transplant.

In certain embodiments, the invention provides compositions and methods for the prophylaxis and/or treatment of mucositis, proctitis and erythemia in a subject who has received, or who is going to receive, mucatoxic chemotherapy with mucositis inducing agents.

In certain embodiments, the invention provides methods and compositions for preventing and/or treating mucositis in a subject who presents with head and/or neck cancer which has been, or which is going to be, treated with radiation therapy with or without adjuvant chemotherapy.

In certain embodiments, the mucositis ,proctitis and erythemia is caused by a subject being exposed to a chemical insult, a biological insult, radiation or a combination thereof. Radiation exposure may result from radiation therapy, for example -6chemotherapy, radiotherapy or the like, or may result from accidental radiation exposure or exposure to radiation following a terrorist attack The compositions, methods and uses of the present invention have further utility in relation to administration to subjects prior to, or following, space travel in order to prevent, treat or ameliorate mucositis, proctitis and erythemia. in certain embodiments, the methods or uses of the invention are performed prior to the subject being subjected to the insult, wherein said insult may induce or cause the progression of mucositis, proctitis and erythemia.

In further embodiments, the methods or uses of the invention may be performed after exposure of the subject to the insult, but prior to the onset and development of mucositis, proctitis and erythemia in the subject.

In yet further embodiments, the methods or uses of the invention may be performed on a subject after the development of mucositis,proctitis,and erythemia in the subject.

The compositions and methods of the present invention can also be used in combination with other therapies to prevent and/or treat mucositis,proctitis or erythemia. . Non-limiting examples of such further therapeutic agents include laser therapy, cryotherapy, antibiotics, cytokine-based therapies such as palifermin (brand name Kepivance, Amgen) which is a human keratinocyte growth factor (KGF), and other cytokine modulators of inflammation, such as IL-11, TGF and GM-CSF.

The compound Kinetin of the present invention can be used in the preparation of a combined medicament comprising at least one compound of the present invention along with a chemotherapeutic agent.

Chemotherapeutic agents suitable for use along with the compositions of the present invention include one or more other anti-tumour substances, for example those selected from mitotic inhibitors, such as vinblastine; alkylating agents, such as cisplatin, carboplatin, and cyclophosphamide, inhibitors of microtubule assembly, such as paclitaxel or other taxanes; anti-metabolites, such as 5-fluorouracil, - 7capecitabine, cytosine arabinoside and hydroxyurea; intercalating antibiotics, such as, adriamycin and bleomycin; immunostimulants, such as trastuzumab; DNA synthesis inhibitors, such as, gemcitabine, enzymes, such as asparaginase; topoisomerase inhibitors, such as etoposide; biological response modifiers, such as interferon, and anti-hormones, for example, antioestrogens, such as tamoxifen, or antiandrogens, such as (4'-cyano-3-(4-fluorophenylsulphonyl )-2-hydroxy-2 -methyl-3' -(trifluoromethyl)propionanilide and other therapeutic agents and principles as described in, for example, DeVita, V. T„ Jr., Hellmann, S,, Rosenberg, S. A.; in: Cancer: Principles & Practice of Oncology, 5th ed., Lippincott-Raven Publishers (1997).

Methods for the administration of such a combined medicament may further be provided by the present invention. In certain embodiments, the compound Kinetin of the present invention and the chemotherapeutic agent are provided sequentially, simultaneously or separately by different routes of administration. Further, said compounds and chemotherapeutic agent may be in the same or different forms, for example a solid and a liquid. Such methods can comprise the simultaneous administration of the compounds of the present invention along with the chemotherapeutic agent. In certain embodiments, the compounds of the present invention may be administered to the subject sequentially with the chemotherapeutic agent. Where they are administered sequentially, in certain embodiments, the compounds of the present invention may be administered prior to the chemotherapeutic agent. In certain further embodiments, the compounds of the present invention may be administered following administration of the chemotherapeutic agent. In certain embodiments, the chemotherapeutic agent is provided separately to the compounds of the present invention.

In certain embodiments, the chemotherapeutic agent and compound Kinetin of the present invention are co-administered. Co-administration means that these components may be administered together as a composition, or as part of the same unitary dose. As used herein, the term co-administration can also mean administering the components separately, but as part of the same therapeutic regimen or treatment program. In certain embodiments, the components are administered to a subject at the same time. However, the components may also be administered - 8separately as separate dosages or dosage forms. Where the components are administered separately, the co-administration of the components does not impose a restriction on the timing, frequency, dosage or order of administration of the components.

Without wishing to be bound by theory, the inventor has identified that the therapeutic and/or prophylactic effect of Kinetin in relation to the treatment or prophylaxis of mucositis is due to enhanced expression and protection of RNA from transcriptional damage following radiation exposure or pre-sensitises the cell to oncoming radiation exp An Evaluation of Kinetin for the Treatment of Radiation Induced Oral Mucositis in Hamsters.

Methods Forty-eight (48) Golden Syrian hamsters were prospectively randomized into nine equally sized groups. On study day 0, mucositis was induced on the left cheek pouch mucosa of the hamsters by a single dose of radiation administered at a dose of 40 Gy. Test articles were given by topical application to the left cheek pouch, and either twice on the day or radiation 2 and 5 hours prior to radiation or once daily beginning on the day before radiation (Day -1) and continuing until Day 20. Animals activity and weight were evaluated daily. Beginning on day 6 and continuing on alternate days for the duration of the study, oral mucositis was evaluated using a standard scoring six point scale The number of days of ulcerative mucositis was evaluated using a Chi-squared test of scores of >3 throughout the study, and the individual daily group scores were assessed with a Rank Sum Test.

Results Mucositis was favorably impacted in the groups that received Kinetin, while the group receiving vehicle only showed no alteration in the levels of mucositis when compared to untreated controls. Control animals had mucositis scores of 3 or higher on 42.7% of animal days evaluated (number of animals multiplied by the number of days evaluated). Animals in groups treated with Kinetin had mucositis scores of 3 or higher on 19.8% to 25% of animal days evaluated. -9KEY WORDS Mucositis, Kinetin, animal models, antioxidant, inflammation, mucosa, epithelia, ulceration, ionizing radiation INTRODUCTION Background Oral ulcerative mucositis is a common, painful, dose-limiting toxicity of chemotherapy and radiation therapy for cancer (4-6). The disorder is characterized by breakdown of the oral mucosa and results in the formation of ulcerative lesions. In granulocytopenic patients, the ulcerations that accompany mucositis are frequent portals of entry for indigenous oral bacteria often leading to sepsis or bacteremia. Mucositis occurs, to some degree, in more than one third of patients receiving antineoplastic drug therapy. The frequency and severity are significantly greater among patients who are treated with induction therapy for leukemia or with many of the conditioning regimens for bone marrow transplant. Among these individuals, moderate to severe mucositis is not unusual in more than three-quarters of patients. Moderate to severe mucositis occurs in virtually all patients who receive radiation therapy for tumors of the head and neck and typically begins with cumulative exposures of 15 Gy and then worsens as total doses of 30 Gy or more are reached Lesions persist throughout the course of radiation therapy and usually spontaneously resolve about 3 weeks following the completion of treatment. Clinically mucositis develops in stages. Early mucosal changes are noted within days of the start of radiation therapy or the administration of chemotherapy. The mucosa becomes erythematous and may, in the case of radiotherapy, demonstrate areas of mild superficial sloughing of epithelium. At this time, patients often complain of slight sensitivity, not dissimilar to what is experienced after a food burn. This is usually easily controlled with topical palliative agents or acetaminophen or NSA1D level analgesics. As mucositis progresses, the epithelium becomes atrophic and ultimately ulcerates. In patients getting stomatotoxic chemotherapy, ulceration begins around 6 days after drug administration. In individuals getting radiation therapy, ulceration is seen at cumulative doses of 20Gy to 30Gy. Once ulcers form then often spread. Individual ulcers coalesce. Symptoms increase exponentially and patients usually require opioid analgesics. The duration of ulceration varies. Those induced by chemotherapy typically peak at 10 to 14 days after infusion and then resolve by day - 1021. In contrast, radiation-induced mucositis is protracted, lasting throughout the usual seven week course of radiation and then not healing (in most cases) for an additional two to three weeks.

Despite its frequency, severity and impact on symptoms and patients’ ability to tolerate cancer treatment, there is currently, only one approved pharmaceutical/biological for the prevention or treatment for oral mucositis. Kepivance (KGF-1, palifermin) was approved for a mucositis indication in patients with hematologic malignancies receiving stem cell transplants.

Standard therapy for mucositis is predominantly palliative. Available agents include topical analgesics such as viscous lidocaine, barrier devices such as GelClair, or rinses such as Caphasol. Systemic analgesics are used for symptom control and antibiotics are used to control secondary infection and mucositis-related bacteremias and sepsis.(4-7). Given the enormity of the unmet clinical need, active studies are ongoing to provide effective interventions for mucositis. The majorities of these are mechanistically based and include biologicals and pharmaceuticals that target the specific pathways thought to be of etiologic importance in the pathogenesis of the condition These include mucosal protective agents, antibiotics, and growth factors such as transforming growth factor (TGF), Interleukin-11 (IL-11), granulocytemacrophage colony stimulating factor (GM-CSF), keratinocyte growth factor (KGF), L-glut.amine, and N-acetyl-cysteine (8-17).

The complexity of mucositis as a biological process has only been recently appreciated. It has been suggested that the condition represents a sequential interaction of oral mucosal cells and tissues, pro-inflammatory cytokines and local factors such as saliva and the oral microbiota. While epithelial degeneration and breakdown ultimately result in mucosal ulceration, it appears that the early changes associated with radiation-induced mucosal toxicity occur within the endothelium and connective tissue of the submucosa. Electron microscopic evaluation of mucosa within 1 week of radiation shows damage to both endothelium and connective tissue, but not epithelium. Such injury is likely mediated by free radical formation. It appears - 11 that the overall mechanism for mucositis development is similar for both radiation and chemotherapy (8, 9).

Acute Radiation Model The acute radiation model in hamsters, developed by the 5 Principle investigator, has proven to be an accurate, efficient and cost-effective technique to provide a preliminary evaluation of anti-mucositis compounds (8, 9). The course of mucositis in this model is well defined and results in peak scores approximately 16-18 days following radiation. The acute model has little systemic toxicity, resulting in few hamster deaths and permitting the use of small groups (N=8) for initial efficacy studies. It has also been used to study specific mechanistic elements in the pathogenesis of mucositis. Molecules that show efficacy in the acute radiation model may be further evaluated in the more complex models of fractionated radiation, chemotherapy, or concomitant therapy.

OBJECTIVE The objective of this study is to evaluate the effect of Kinetin on the progression of radiation induced oral mucositis in Syrian Golden Hamsters.

MATERIAL AND METHODS Study Locations The study was performed at Biomodels’ AAALAC accredited facility in Watertown, MA Approval for this study was obtained from the Biomodels Institutional Animal Care and Use Committee (IACUC). The 1ACUC approval number for this study is 070620-01 Animals Male Golden Syrian hamsters (Charles River Laboratories), aged 5 to 6 weeks, with a mean body weight 81.4 g at study commencement, were used. Animals were individually numbered using an ear punch and housed in groups of 8 animals per cage. Animals were acclimatized for 5 days prior to study commencement. During acclimatization, the animals were observed daily in order to reject animals that presented in poor condition.

Housing - 12The study was performed in animal rooms provided with filtered air at a temperature of 70oF+/-5o F and 50 +/-20% relative humidity. Animal rooms were set to maintain a minimum of 12 to 15 air changes per hour. The room was on an automatic timer for a light - dark cycle of 12 hours on and 12 hours off with no twilight Sterile Bed-OCobsti) bedding was used. Bedding was changed a minimum of once per week Cages, tops, bottles, etc. were washed with a commercial detergent, rinsed and allowed to air dry. Floors were swept daily and mopped a minimum of twice weekly with a commercial detergent. Walls and cage racks were sponged a minimum of once per month with a dilute bleach solution. The temperature and relative humidity were recorded during the study, and the records retained.

Diet Animals were fed with LabDiet® 5053 Rodent Diet and water was provided ad libitum.

Animal Randomization and Allocations Hamsters were randomly and prospectively divided into eleven groups. Each animal was identified by an ear punch corresponding to an individual number. A cage card identified each cage or label marked with the study number, treatment group number and animal numbers.

Mucositis Induction Mucositis was induced using a standardized acute radiation protocol. A single dose of radiation (40 Gy/dose) was administered to all animals on day 0. Radiation was generated with a 160 kilovolt potential (15-ma) source at a focal distance of 50 cm, hardened with a 0.35 mm Cu filtration system. Irradiation targeted the left buccal pouch mucosa at a rate of approximately 2.2 Gy/minute. Prior to irradiation, animals were anesthetized with an intraperitoneal injection of ketamine (160 mg/ml) and xylazine (8 mg/ml). The left buccal pouch was everted, fixed and isolated using a lead shield. - 13 Study Design Forty-eight (48) male Syrian Golden Hamsters were randomly assigned to six (6) groups of eight (8) animals each, On study day 0, each hamster was given an acute radiation dose of 40 Gy directed to their left buccal cheek pouch. This was accomplished by anesthetizing the animals and everting the left buccal pouch, while protecting the rest of the animals with a lead shield. Test articles were given as described in Table 1.

Table 1: Study Design Croup No. Of Animals Treatment Route of Administration and Treatment Schedule Duration of Dosing Volume (mL) 1 8/male Untreated NA NA NA 2 8/male Vehicle (20% ethanol) Topical 3 hours prior to radiation and 1 hour prior to radiation Day 0 0.2mL 3 8/male Kinetin 0.2mg Topical ,2mg 3 hours and 2mg 1 hour prior to radiation Day 0 0 2mL 4 8/male Kinetin 12mg Topical 12mg 3 hours and 12mg 1 hour prior to radiation Day 0 0.2mL s 8/malc Kinctin 20mg Topical 20mg 3 hours and 20mg 1 hour prior to radiation Day 0 0,2mL 6 8/male Kinetin 0.2mg Topical 2mg 3 hours and 2mg 1 hour prior to radiation Days -1 to 20* 0.2mL ^Dosing on day 0 will be performed 2 hours prior to radiation OUTCOME EVALUATION Study endpoints were mucositis score, weight change and survival.

Survival and Weight Change Data Animal deaths were evaluated during the course of the study. In general, deaths are usually attributable to adverse effects associated with anesthesia which typically occur - 14at the time of radiation, or toxicity of the experimental compound. Since weight change is a secondary method to examine potential toxicities of experimental treatments, animals were weighed daily throughout the study.

Mucositis Evaluation To evaluate mucositis severity, animals were anesthetized with an inhalation anesthetic, and the left cheek pouch everted. Mucositis was scored visually by comparison to a validated photographic scale, as shown in Figure 2. The scale ranges from 0 for normal, to 5 for severe ulceration. In descriptive terms, this scale is defined as follows.

Table 2: Mucositis Scoring Table Score: Description: 0 Pouch completely healthy. No erythema or vasodilation 1 Light to severe erythema and vasodilation. No erosion of mucosa 2 Severe erythema and vasodilation. Erosion of superficial aspects of mucosa leaving denuded areas. Decreased stippling of mucosa. 3 Formation of off-white ulcers in one or more places. Ulcers may have a yellow/gray due to pseudomembrane. Cumulative size of ulcers should equal about % of the pouch. Severe erythema and vasodilation. 4 Cumulative size of ulcers should equal about '/a of the pouch. Loss of pliability. Severe erythema and vasodilation 5 Virtually all of pouch is ulcerated. Loss of pliability (pouch can only partially be extracted from mouth.

A score of 1-2 is considered to represent a mild stage of the disease, whereas a score of 3-5 is considered to indicate moderate to severe mucositis. Following visual scoring, a photograph was taken of each animal’s mucosa using a standardized technique. At the conclusion of the experiment, all films were developed and the photographs randomly numbered. At least two independent trained observers graded the photographs in blinded fashion using the above described scale {blinded scoring). - 15 Clinical Significance of the Mucositis Scoring System The clinical mucositis score of 3 in hamsters indicates the presence of an ulcer (see Figure 2). Ulceration is the point in the development of mucositis where the physical integrity of the oral mucosa is breached. In the clinic, a patient presenting with severe oral ulcerations may require hospitalization for analgesic, narcotic and/or antibiotic therapies or fluid support. The average cost to the healthcare system is significant. Advanced mucositis in humans (with ulcerative sores) often requires the interruption of therapy for patients receiving radiation and, if sepsis occurs, these patients risk death. A therapeutic that significantly reduced the time that a patient with oral mucositis had ulcers would be of great value to the clinician. In the hamster model of oral mucositis, the duration of scores of 3 or greater is used as a primary measurement of efficacy in mucositis treatment.

Analysis of Efficacy of Compounds in Treatment of Mucositis The mean group mucositis scores were compared to the control group in each experiment. This comparison provided an effective graphic representation through which the relative effectiveness of each compound was initially determined. Since the mucositis score of 3 has such great clinical importance, the cumulative number of days that an animal has a score of 3 or greater was determined. The significance of group differences in scores of 3 or greater was determined using Chi-squared (χ2) difference analysis for cumulative scores over the course of the entire study.

Administration of Test Articles For topical administration, hamsters were dosed by placing a needless 1 mL syringe into the left cheek pouch and administering the appropriate volume of test article RESULTS AND DISCUSSION Survival No animal deaths were noted during this study Weight Change (Figures 3 & 4).

No statistically significant differences were seen (p=0.409). The mean daily percent weight gains for each group are shown in Figure 3. The untreated control group had a - 16mean weight gain of 77% of their starting weight during the course of the study. The 20% ethanol control group had a mean weight gain of 76%. The groups treated with Kinetin on day 0 only had mean weight gains of 75%, 79% and 67% respectively for the 0.2 mg, 12 mg and 20 mg groups. The group treated with Kinetin from day -1 to day 20 had a mean weight gain of 80% during the study, To evaluate the significance of these differences, the mean area under the curve (AUC) was calculated for each animal from the percent weight gain data. To compare the effects of treatment with oltipraz treated groups to controls, a one-way ANOVA test was used to compare all groups. The mean percent weight gain AUC data is shown in Figure 4 Mucositis (Figures 5 & 6, Tables 3 & 4) The mean daily mucositis scores for all study groups are shown in Figure 5. All groups treated with Kinetin showed significant reductions in oral mucositis.

Group Days •-3 Days <3 Total Days % Days >=3 Chi So UtUreaStc V P Value ChiSq MS EtOH control P Value Untreated 82 no 182 42.7 - • * - 2O'-t Lttiai’cn Vehicle Control bid day 0 110 1S2 42.7 0.0106 o.s is - - Kinetin Q J mg bid day ΰ 3e iS4 1 Uz W.B _ t 2-.· - .. i!.l — Kinetin 12 mg bid day 0 8 144 j .25.0 _ ύ _ ? - Ki n eti n 2$ rag w 25,0 12.6S40 bid day ¢1 Kinetin 0.2 ns 44 i-«a 192 1S. 17 Kjh I 16.1710 54461 Table 3: Chi-squared analysis of number of animal days with a mucositis score of 3 or higher.

To examine the levels of clinically significant mucositis, as defined by presentation 20 with open ulcers (score the total number of days in which an animal exhibited an elevated score was summed and expressed as a percentage of the total number of days scored for each group. Statistical significance of observed differences was calculated - 17using chi-square analysis. Significant improvement is shown in Red type.

Bay Group Comparison 6 8 10 12 14 18 18 20 22 24 2S ja Untreated vs fcsOH control· a, m O.ZJS 0,·« r 0.S3S O.iSB s.sss 0388 0.SS4 0.232 o.®e 0..S03 o.eto Untreated vi Kin etin ύ. Ϊ mg bid ii3y 6 0.985 0-.3/1 «6.001 iSi-Lii 0...-0 ft see 0.135 C.W0 0..273 0. ? 2*3 Untreated vs Kinetic 15 mg bid Bay 0 ΰ .1*65 O.LiL’b 01. i 65 0,054 O.SiM· 0.. ΐ3ίϊ a.o® 0.032 Untreated vs Kin etin 26 mg bid Bay 0 cress 0.3/ 1 £ _». j ό O.iSfi u.:isa S 304 0.13S fi.s 10 0.212 0.10' Untreated vs Kin etin 0,2 mg Bays -t to 20 O.'SSii 1 0,fi5i 0,135 0.2 S3 0.(:-5 Table 4. Evaluation of Daily Mucositis Score Variations - Comparison with 5 Untreated Controls.

The significance of group differences observed in daily mucositis scores was determined using the Mann-Whitney rank sum test. This nonparametric statistic is appropriate for the visual mucositis scoring scale. The p values for each calculation are shown. Significant improvements are shown in RED.

Bey Group Csmparfson 6 18 12 14 16 13 20 2*2 24 2e 25) etOW Vehicle • ·· Kinetin 0.2 >,·!.; bid Bay 0 0,//·. 0.505 0.071 —-S • '-Ll ’ 0 .;.JU - ΓΗ- EtOH Vehtote vs Kinetin 12 mg bid Day 0 0./74 G.7i-i 0.507 ί»,3δ·ί a. ' - 3 - > ·< CtOH Vehicle vs Kin etin 26 mg hid Bay it 0. 774 O.5SS 0. rtS 0.00; il.C&l 0.30; 07:30 0,077 Sill 0 J ttose «Mi EtOH Vehicle vs Kin etin 0,2 eng Bays -1 t© 26 0.774 0.55S Q.S01 0.061 0.072 0.04¾ 0.554 0.07 § § ή 0 OiM Table 5. Evaluation of Daily Mucositis Score Variations — Comparison with Vehicle Controls The significance of group differences observed in daily mucositis scores was 15 determined using the Mann-Whitney rank sum test. This nonparametric statistic is appropriate for the visual mucositis scoring scale. The p values for each calculation are shown. Significant improvements are shown in RED. - 18 Ulcerative Severity Analysis The significance of the differences between the vehicle group and the treated groups was assessed in 2 ways, first by the comparison of the number of days with an ulcer (i.e. a score of 3 or higher) using a chi-squared (χ2) test. The results of the analysis of animal days with a score of 3 or higher are shown in Table 3 and Figure 5. Untreated hamsters had a score of 3 or higher on 82 or 192 animal days evaluated (42.7%). Animals dosed with 20% ethanol vehicle had scores of 3 or higher on 82 of 192 animal days evaluated (42.7%).

All the groups treated with Kinetin showed significant reduction in the number of animals with a mucositis score of 3 or higher, with significant reductions for all groups when compared to both untreated and vehicle controls (p<0 001 for all comparisons). The group treated with Kinetin at 0.2 mg on day 0 had the fewest animal days with a score of 3 or higher (19.8%). The other 2 groups treated on day 0 had a mucositis score of 3 or higher on 25% of animal days evaluated. The group treated with Kinetin on days -1 to 20 had a mucositis score of 3 or higher on 22.9% of animal days evaluated.

Figure 2: Mucositis Score Scale.

Figure 3. Mean percent weight change: Animals were weighed daily, the percent weighs change from day 0 was calculated, and group means and standard errors of the mean (SEM) calculatedfor each day.

Figure 4. Mean Percent Weight Change AUC The area under the curve (AUC) was calculated for the percent weight change exhibited by each animal in the study. This calculation was made using the trapezoidal rule transformation. Group means were calculated and are shown with error bars representing SEM for each group. A One Way ANOVA was conducted to compare differences between these groups. No significant differences were noted between any groups (p=0.409). - 19Figure 5: Mean Daily Mucositis Scores. Mean group mucositis scores were obtained every other day. Error bars represent the standard error of the mean (SEM). (A) Topically Treated Groups (B) Orally Dosed Groups.

Figure 6: Duration of Severe Mucositis Number of days with mucositis scores of >3. To examine the levels of clinically signif icant mucositis, as defined by presentation with open ulcers (score :>3), the total number of days in which an animal exhibited an elevated score was summed and expressed as a percentage of the total number of days scored for each group.

JO Statistical significance of observed differences was calculated using chi-square analysis. Significant improvements are denoted with an asterisk.

Rank Sum Analysis Further analysis of the data was performed using the Mann-Whitney Rank-sum analysis on the mucositis scores for individual days comparing treated groups to the control groups. In this analysis the data for each group at each time-point is compared with the control group. The results of this analysis are shown in Tables 4, 5.

There were no significant differences between the vehicle treated group and the untreated control group.

The groups treated with Kinetin were similar to one another, showing a pattern of significant improvements on days 10-16 when compared to the untreated controls, and improvements at these timepoints as well as days 22 to 28 when compared to the vehicle controls. The group treated with Kinetin at 0.2 mg on day 0 showed significant reductions in mucositis scores when compared to either the untreated controls (p<0.001 on days 10, 12 and 14, and p-0.004 on day 16), or the vehicle treated controls (p<0.001 on day 12, p=0.002 on day 14, p=0.011 on days 16, 22 and 24, and p-0.025 on day 28). The group treated with Kinetin at 12 mg on day 0 showed significant reductions in mucositis scores when compared to either the untreated controls (p<0,001 on days 10 and 14, p=0.002 on day 12 and p—0.01 I on day 28), or the vehicle treated controls (p=0.004 on day 12, p=0.011 on days 14, 22 and 24, p=0.015 on day 26 and p<0.001 on day 28), The group treated with Kinetin at -2020 mg on day 0 showed significant reductions in mucositis scores when compared to either the untreated controls (p<0.001 on days 10 and 14, and p=0.003 on day 12), or the vehicle treated controls (p=0.007 on day 12, p=0.011 on day 22 and p=0.019 on day 28). The group treated with Kinetin at 0.2 mg on days -1 to 20 showed significant reductions in mucositis scores when compared to either the untreated controls (p=0.016 on day 10, p=0.030 on day 12, p=0.001 on day 14, p=0 016 on day 16 and p=4),038 on day 26), or the vehicle treated controls (p=0.046 on day 16, p=0.011 on day 22, p-OO35 on day 24, p=0.004 on day 26 and p=0.005 on day 28).

Conclusions Based on observed survival and weight changes, the test materials appeared to be well tolerated 2. Kinetin was effective at reducing the duration of severe oral mucositis at all doses and schedules evaluated in this study (p<0.001 for all comparisons). 3. Kinetin was effective at reducing the severity of oral mucositis at all doses and schedules evaluated in this study both before and after the peak of mucositis.

Claims

J. A method for prophylaxis and/or treatment of mucositis, Proctitis and erythemia the method comprising the steps of: - providing a therapeutically effective amount of a composition comprising N6-Furfuryadenine (ΚΓΝΕΤΙΝ) or an analogue, derivative, metabolite, prodrug, solvate or pharmaceutically acceptable salt thereof; and administering the composition to a subject.
2. A method as claimed in claim 1 wherein the composition is administered to a subject after the subject has undergone treatment comprising the administration of at least one of a chemotherapeutic agent, a radiation therapy or a combination thereof.
3. A method as claimed in claim 1 wherein the composition is administered to a subject prior to the subject undergoing treatment comprising the administration of at least one of a chemotherapeutic agent, a radiation therapy or a combination thereof.
4. A method as claimed in claim 2 wherein the composition is administered to the subject after the subject has undergone treatment comprising the administration of at least one of a chemotherapeutic agent and/or radiation therapy but wherein said compositions is administered to the subject prior to the determination in the subject of at least one symptom indicative of mucositis, proctitis and or erythemia.
5. A method as claimed in claim 2 wherein the composition is administered to the subject after the diagnosis of the subject with at least one symptom indicative of mucositis, proctitis and or erythemia.
6. A method as claimed in claim 1 wherein the composition is administered to the subject along with the administration of a chemotherapeutic agent, a radiation therapy or a combination thereof. -227. A method as claimed in claim 6 wherein the composition and the chemotherapeutic agent, radiation therapy or combination thereof are administered to the subject sequentially.
7. 8. A method as claimed in claim 6 wherein the composition and the chemotherapeutic agent, radiation therapy or combination thereof are administered to the subject separately.
8. 9. A method as claimed in claim 6 wherein the composition and the chemotherapeutic agent, radiation therapy or combination thereof are administered to the subject simultaneously
9. 10 A method as claimed in anyone of the preceding claims wherein the form of mucositis is selected from at least one of the group consisting of enteritis, oropharyngeal mucositis, stomatitis and proctitis.
10. 11. A method as claimed in anyone of the preceding claims wherein the mucositis is caused by the subject being exposed to at least one of a chemical insult, a biological insult, a radiation insult or a combination thereof, wherein said insult results in the onset of mucositis.
11. 12. A method as claimed in claim 1 wherein the compound of this patent is administered simultaneously both by oral dosage and by mouth rinse to effect treatment of mucositis.
12. 13. A method as claimed in claim 12 wherein the compound of this patent Kinetin is formulated for mouth rinse in honey or in agave syrup to both mask taste and enhance healing properties.
13. 14. A method as claimed in claim 12 wherein the active compound of this patent Kinetin is administered by suppository to enhance patient compliance and to overcome difficulties with swallowing. -2315. A method as claimed in Claim 1 wherein the compound of this patent Kinetin is formulated in a trans-buccal delivery system to effect rapid delivery.
14. 16. A method as claimed in anyone of the preceding claims wherein the composition is administered to the subject in the form of an oral rinse, in liquid form, in a capsule, in a tablet, in an injection or as a suppository.
15. 17. A method as claimed in claim 16 wherein the composition is administered in the form of an oral rinse
16. 18. Use of a composition comprising at least one compound selected from the group consisting of 5-[2-pyrazinyl]-4-methyl-l,2-dithiol-3-tbione or an analogue, derivative, metabolite, prodrug, solvate or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the treatment and/or prophylaxis of mucositis.
17. 19. Use of a composition as claimed in claim 19 wherein the composition further comprises a chemotherapeutic agent.
18. 20 Use of a composition as claimed in claim 19 or 20 wherein the metabolite of 5-[2-pyrazinyl]-4--methyl-l,2-dithiol-3-thione is the pyrrolopyrazine derivative metabolite 3.
19. 21. Use of a composition as claimed in claim 19 or 20 wherein the derivative of 5-[2-pyrazinyl]-4-methyl-l,2-dithiol-3-thione is anethole trithione.
20. 22. Use of a composition as claimed in anyone of claims 19 to 22 wherein the composition further comprises carboxymethyl cellulose,
21. 23. Use of a composition as claimed in anyone of claims 19 to 23 wherein the composition further comprises a sulphur-containing amino acid. -2424. Use of a composition as claimed in claim
22. 24 wherein the sulphur containing amino acid is cysteine or an analogue, salt or solvate thereof.
23. 25 Use of a composition comprising 5-[2-pyrazinyl]-4-methyl-l,2-dithiol-3thione or an analogue, salt, derivative, solvate or metabolite thereof in the preparation of a medicament in the form of an oral rinse for administration to a subject prior to the subject undergoing treatment with a chemotherapeutic agent, radiation therapy or combination thereof for the prophylaxis and/or treatment of mucositis.
24. 26. Use of a composition comprising 5-[2-pyrazinyl]-4-methyl-l,2-dithiol-3thione or an analogue, salt, derivative, solvate or metabolite thereof in the preparation of a medicament in the form of an oral rinse for administration to a subject following the subject undergoing treatment with a chemotherapeutic agent, radiation therapy or combination thereof for the prophylaxis and/or treatment of mucositis,
25. 27. A pharmaceutical composition comprising 5-[2-pyrazinyl]-4-methyl-],2dithiol-3 -thione or an analogue, derivative, metabolite, prodrug, solvate or a pharmaceutically acceptable salt thereof along with at least one pharmaceutically acceptable carrier, excipient or diluent.
26. 28. A pharmaceutical composition as claimed in claim 28 wherein the composition further comprises a chemotherapeutic agent.
27. 29. A pharmaceutical composition as claimed in claim 28 or 29 wherein the metabolite of 5-[2-pyrazinyI]-4-methyI-l,2-dithiol-3-thione is the pyrrolopyrazine derivative metabolite 3.
28. 30. A pharmaceutical composition as claimed in claim 28 or 29 wherein the derivative of 5-[2-pyrazinyl]-4-methyl-l,2-dithiol-3-thione is anethole trithione. -25 3 1. A pharmaceutical composition as claimed in anyone of claims 28 to
29. 31 wherein the composition further comprises carboxymethyl cellulose.
30. 32. A pharmaceutical composition as claimed in anyone of claims 28 to 32 wherein the composition further comprises a sulphur-containing amino acid
31. 33. A pharmaceutical composition as claimed in claim 33 wherein the sulphurcontaining amino acid is cysteine or an analogue, salt or solvate thereof
32. 34 A pharmaceutical composition as claimed in anyone of claims 28 to 34 for use in the treatment and/or prophylaxis of mucositis.
33. 35. A method for prophylaxis and/or treatment of mucositis, the method comprising the steps of: providing a therapeutically effective amount of a composition comprising at least one cytokinin compound, or a pharmaceutically acceptable salt or solvate thereof; and administering the composition to a subject in need of such treatment.
34. 36. A method as claimed in claim 36 wherein the cytokinin compound is N 6 isopentenyl adenosine or an analogue, derivative, metabolite, prodrug, solvate or salt thereof
35. 37. A method as claimed in claim 36 wherein the cytokinin compound is N 6 benzyl adenosine or an analogue, derivative, metabolite, prodrug, solvate or salt thereof.
36. 38 . A method as claimed in anyone of claims 36 to 38 wherein the subject has undergone treatment with a chemotherapeutic agent, a radiation therapy or a combination thereof - 2639. A method as claimed in anyone of claims 36 to 38 wherein the composition is administered to the subject prior to the subject undergoing treatment with the chemotherapeutic agent, radiation therapy or 5 combination thereof.
37. 40. A method as claimed in claim 39 wherein the composition is administered to the subject after the subject has undergone treatment with the chemotherapeutic agent and/or radiation therapy but prior to the diagnosis 10 of symptoms indicative of mucositis in the subject.
38. 41. A method as claimed in claim 39 wherein the composition is administered to the subject after the diagnosis of symptoms indicative of mucositis in the subject.
39. 42. A method as claimed in anyone of claims 36 to 38 wherein the composition is administered with a chemotherapeutic agent, a radiation therapy or a combination thereof. 20
40. 43, A method as claimed in claim 43 wherein the composition and the chemotherapeutic agent, the radiation therapy or the combination thereof are administered to the subject sequentially.
41. 44. A method as claimed in claim 43 wherein the composition and the 25 chemotherapeutic agent, the radiation therapy or the combination thereof are administered to the subject separately.
42. 45 A method as claimed in claim 43 wherein the composition and the chemotherapeutic agent, the radiation therapy or the combination thereof 30 are administered to the subject simultaneously. -2746. A method as claimed in anyone of claims 36 to
43. 46 wherein the mucositis is selected from at least one of the group consisting of enteritis, esophagitis, oropharyngeal mucositis, stomatitis and proctitis.
44. 47. A method as claimed in anyone of claims 36 to 47 wherein the mucositis is caused by the subject being exposed to at least one of a chemical insult, a biological insult, a radiation insult or a combination thereof wherein said insult results in the onset of mucositis.
45. 48. A method as claimed in any claims 36 to 48 wherein the composition further comprises carboxymethyl cellulose.
46. 49. A method as claimed in anyone of claims 36 to 49 wherein the composition is administered to the subject in the form of an oral rinse, in liquid form, in a capsule, in a tablet, in an injection or as a suppository.
47. 50. Use of a composition comprising at least one cytokinin compounds or a pharmaceutically acceptable salt or solvate there of in the preparation of a medicament for the treatment and/or prophylaxis of mucositis
48. 51. Use of a composition as claimed in claim 51 wherein the cytokinin compound is N 6 -isopentenyl adenosine or an analogue, derivative, metabolite, prod rug or salt thereof
49. 52. Use of a composition as claimed in claim 51 wherein the cytokinin compound is N 6 -benzyl adenosine or an analogue, derivative, metabolite, prod rug or salt thereof
50. 53. Use of a composition as claimed in any of claims 51 to 53 wherein the composition comprises a chemotherapeutic agent.
51. 54. Use of a composition as claimed in any of claims 51 to 54 wherein the composition further comprises carboxymethyl cellulose. -2855. A pharmaceutical composition comprising at least one cytokinin compound or a pharmaceutically acceptable salt or solvate thereof along with a pharmaceutically acceptable carrier.
52. 56. A pharmaceutical composition as claimed in claim 56 wherein the cytokinin compound is N 6 -isopentenyl adenosine or an analogue, derivative, metabolite, prodrug, solvate or salt thereof
53. 57. A pharmaceutical composition as claimed in claim 56 wherein the cytokinin compound is N 6 -benzyl adenosine or an analogue, derivative, metabolite, prodrug, solvate or salt thereof.
54. 58. A pharmaceutical composition as claimed in anyone of claims 56 to 58 wherein the composition comprises a chemotherapeutic agent.
55. 59. A pharmaceutical composition as claimed in anyone of claims 56 to 59 wherein the composition further comprises carboxymethyl cellulose.
56. 60. A pharmaceutical composition as claimed in anyone of claims 56 to 60 for use in the treatment and/or prophylaxis of mucositis.
57. 61. A combined medicament comprising at least one cytokinin compound or a pharmaceutically acceptable salt or solvate thereof and 5-[2-pyrazinyl]-4methyl-l,2-dithioI-3-thione or an analogue, derivative, metabolite, prod rug, solvate or a pharmaceutically acceptable salt thereof.
58. 62. A method of reducing and/or preventing weight loss in a subject undergoing cancer treatment, the method comprising the steps of providing a therapeutically effective amount of a composition comprising at least one compound selected from the group consisting of 5-[2-pyrazinyl]-4-methyll,2-dithiol-3-thione or an analogue, derivative, metabolite, prodrug, -29solvate or a pharmaceutically acceptable salt thereof; and administering the composition to the subject.
59. 63. A method as claimed in claim 63 wherein the subject has undergone treatment with a chemotherapeutic agent, a radiation therapy or a combination thereof
60. 64. A method as claimed in claim 63 wherein the composition is administered to a subject prior to the subject undergoing treatment with a chemotherapeutic agent, a radiation therapy or a combination thereof
61. 65. A method as claimed in claim 64 wherein the composition is administered to the subject after the subject has undergone treatment with the chemotherapeutic agent, radiation therapy or combination thereof but prior to onset of weight loss in the subject.
62. 66 A method as claimed in claim 64 wherein the composition is administered to the subject after onset of weight loss in the subject.
63. 67. A method as claimed in claim 63 wherein the composition is administered with a chemotherapeutic agent, a radiation therapy or a combination thereof.
64. 68. A method as claimed in claim 68 wherein the composition and the chemotherapeutic agent, radiation therapy or combination thereof are administered to the subject sequentially.
65. 69. A method as claimed in claim 68 wherein the composition and the chemotherapeutic agent, radiation therapy or combination thereof are administered to the subject separately. -3070. A method as claimed in claim 68 wherein the composition and the chemotherapeutic agent, radiation therapy or combination thereof are administered to the subject simultaneously.
66. 71 A method as claimed in anyone of claims 63 to 7 J wherein the weight loss is caused the subject being exposed to at least one of a chemical insult, a biological insult, a radiation insult or a combination thereof
67. 72 A method as claimed in anyone of claims 63 to 72 wherein the metabolite of 55~[2-pyrazinyl]-4-methyl-l,2-dithiol-3-thione is the pyrrolopyrazine derivative metabolite 3.
68. 73. A method as claimed in anyone of claims 63 to 72 wherein the derivative of 5-[2-pyrazinyl]-4-methyl-l,2-dithiol-3-thione is anethole trithione.
69. 74. A method as claimed in anyone of claims 63 to 74 wherein the composition further comprises carboxymethyl cellulose.
70. 75. A method as claimed in anyone of claims 63 to 75 wherein the composition further comprises a sulphur-containing amino acid.
71. 76. A method as claimed in claim 76 wherein the sulphur-containing amino acid is cysteine or an analogue or derivative thereof
72. 77. A method as claimed in anyone of claims 63 to 77 wherein the composition is administered to the subject in the form of an oral rinse, in liquid form, in a capsule, in a tablet, in an injection or as a suppository.
73. 78. Use of a composition comprising at least one compound selected from the group consisting of 5-[2-pyrazinyl]-4-methyl-l,2-dithiol-3-thione or an analogue, derivative, metabolite, prodrug, solvate or a pharmaceutically acceptable salt thereof in the preparation of a medicament for reducing and/or preventing weight loss in a subject undergoing cancer treatment. - 31
74. 79. Use of a composition as claimed in claim 79 wherein the composition further comprises a chemotherapeutic agent.
75. 80. Use of a composition as claimed in claim 79 or 80 wherein the metabolite of 5-[2-pyrazinyl]-4-methyl-l,2-dithiol-3-thione is the pyrrolopyrazine derivative metabolite 3.
76. 81. Use of a composition as claimed in claim 79 or 80 wherein the derivative of 5-[2-pyrazinyl]-4-methyl-l,2-dithiol-3-thione is anethole trithione.
77. 82. Use of a composition as claimed in anyone of claims 79 to 82 wherein the composition further comprises carboxymethyl cellulose.
78. 83. Use of a composition as claimed in anyone of claims 79 to 83 wherein the composition further comprises a sulphur-containing amino acid
79. 84. Use of a composition as claimed in claim 84 wherein the sulphurcontaining amino acid is cysteine or an analogue, salt or solvate thereof.
80. 85. A method of prophylaxis and/or treatment of cachexia, the method comprising the steps of: providing a therapeutically effective amount of a composition comprising at least one compound selected from the group consisting of 5-[2-pyrazinyl]-4-methyl-l,2-dithiol-3-thione or an analogue, derivative, metabolite, prodrug, solvate or a pharmaceutically acceptable salt thereof; and administering the composition to a subject in need of such treatment.
81. 86. A method as claimed in claim 86 wherein the cachexia is caused by at least one of a chemical insult, a biological insult, a radiation insult or a combination thereof. - 3287. A method as claimed in claim 86 or
82. 87 wherein the metabolite of 5-[2pyrazinyl]-4-methyl-l,2-dithiol-3-thione is the pyrrolopyrazine derivative metabolite 3.
83. 88. A method as claimed in claim 86 or 87 wherein the derivative of 5-[2pyrazinyl]-4-methyl-l,2-dithiol-3-thione is anethole trithione.
84. 89. A method as claimed in anyone of claims 86 to 89 wherein the composition further comprises carboxymethyl cellulose.
85. 90. A method as claimed in anyone of claims 86 to 90 wherein the composition further comprises a sulphur-containing amino acid.
86. 91. A method as claimed in claim 91 wherein the sulphur-containing amino acid is cysteine or an analogue, salt or solvate thereof.
87. 92. A method as claimed in anyone of claims 86 to 92 wherein the composition is administered to the subject in the form of an oral rinse, in liquid form, in a capsule, in a tablet, in an injection or as a suppository
88. 93. Use of a composition comprising at least one compound selected from the group consisting of 5-[2-pyrazinyl]-4-methyl-l,2-dithiol-3-thione or an analogue, derivative, metabolite, prodrug, solvate or a pharmaceutically acceptable salt thereof in the preparation of a medicament for prophylaxis and/or treatment of cachexia in a subject.
89. 94. Use of a composition as claimed in claim 94 wherein the metabolite of 5[2-pyrazinyl]-4-methyl-l,2-dithiol-3-thione is the pyrrolopyrazine derivative metabolite 3.
90. 95. Use of a composition as claimed in claim 94 wherein the derivative of 5[2-pyrazinyl]-4-methyl-l,2-dithiol-3-thione is anethole trithione. - 33
91. 96. Use of a composition as claimed in anyone of claims 94 to 96 wherein the composition further comprises carboxymethyl cellulose.
92. 97. Use of a composition as claimed in anyone of claims 94 to 97 wherein the composition further comprises a sulphur-containing amino acid.
93. 98 Use of a composition as claimed in claim 98 wherein the sulphurcontaining amino acid is cysteine or an analogue, salt or solvate thereof
94. 99. Use of a composition as claimed in anyone of claims 94 to 99 wherein the cachexia is caused by the subject being exposed to at least one of a chemical insult, a biological insult, a radiation insult or a combination thereof, wherein said insult results in the onset of cachexia.
95. 100. A pharmaceutical composition as claimed in anyone of claims 28 to 34 for use in the treatment and/or prophylaxis of weight loss and/or cachexia.