DOSING REGIMENS FOR TREATING METAL-MEDIATED CONDITIONS

Abstract

The present disclosure provides safe and effective dosing regimens of metal chelators as treatment for metal overload disorders and, in particular, iron overload and associated conditions.

Claims

1-20. (canceled)

21. A method of reducing the renal toxicity of an SP-420 compound, comprising orally administering to a subject suffering from transfusional iron overload an SP-420 compound three or four times per week no more frequently than every other day, wherein the weekly dose of the SP-420 compound administered would exhibit renal toxicity if administered in seven daily doses.

22. The method of claim 21, wherein the SP-420 compound is administered three times per week.

23. The method of claim 21, wherein the SP-420 compound is administered four times per week.

24. The method of claim 21, wherein the SP-420 compound is administered for a period of at least one week.

25. The method of claim 21, wherein the SP-420 compound is administered for a period of at least one month.

26. The method of claim 21, wherein the SP-420 compound is administered for a period of at least three months.

27. The method of claim 21, wherein the SP-420 compound is administered for a period of at least six months.

28. The method of claim 21, wherein the SP-420 compound is administered for a period of at least one year.

29. The method of claim 21, wherein the SP-420 compound is administered indefinitely.

30. The method of claim 21, wherein the SP-420 compound is SP-420 ((S)-4,5-dihydro-2-[2-hydroxy-4-(3,6-dioxaheptyloxy)phenyl]-4-methyl-4-thiazolecarboxylic acid).

31. The method of claim 21, wherein the SP-420 compound is a pharmaceutically acceptable salt of SP-420 ((S)-4,5-dihydro-2-[2-hydroxy-4-(3,6-dioxaheptyloxy)phenyl]-4-methyl-4-thiazolecarboxylic acid).

32. The method of claim 31, wherein the SP-420 compound is a sodium salt of SP-420 ((S)-4,5-dihydro-2-[2-hydroxy-4-(3,6-dioxaheptyloxy)phenyl]-4-methyl-4-thiazolecarboxylic acid).

33. The method of claim 30, wherein the SP-420 compound is administered three times per week.

34. The method of claim 30, wherein the SP-420 compound is administered four times per week.

35. The method of claim 30, wherein the SP-420 compound is administered for a period of at least one week.

36. The method of claim 30, wherein the SP-420 compound is administered for a period of at least one month.

37. The method of claim 30, wherein the SP-420 compound is administered for a period of at least three months.

38. The method of claim 30, wherein the SP-420 compound is administered for a period of at least six months.

39. The method of claim 30, wherein the SP-420 compound is administered for a period of at least one year.

40. The method of claim 30, wherein the SP-420 compound is administered indefinitely.

41. The method of claim 21, wherein the subject has β-thalassemia.

42. The method of claim 21, wherein the subject has sickle cell anemia, Diamond-Blackfan anemia, sideroblastic anemia, chronic hemolytic anemia, off-therapy leukemia, or myelodysplastic syndrome.

43. The method of claim 21, wherein the subject has myelodysplastic syndrome.

44. The method of claim 21, wherein the subject has had a bone marrow transplant.

Description

4. BRIEF DESCRIPTION OF THE FIGURES

[0016] FIG. 1A-1B show representative KIM-1 data for rats in Example 2 receiving SP-420 every day (FIG. 1A) or every other day (FIG. 1B).

[0017] FIG. 2 shows KIM-1 strips for the rats in Example 2 after having been given a cumulative dose of SP-420 of 2600 mg/kg.

[0018] FIG. 3 shows KIM-1 strips for the rats in Example 2 that had been administered 162.5 mg/kg SP-420 daily for 16 days at the end of the dosing period (left side) and five days post administration (right side).

[0019] FIG. 4 shows a 24-hour urine sample of a rat administered SP-420 daily at a dose of 200 mg/kg/day.

[0020] FIG. 5 shows average KIM-1 data for the rats of Example 2.

5. DETAILED DESCRIPTION

5.1 Metal-Mediated Conditions

[0021] Metal ions are critical to the proper functioning of living systems. Ions such as Fe.sup.3+, Zn.sup.2+, Cu.sup.2+, Ca.sup.2+, and Co.sup.3+, to name but a few, can be found in the active sites of over a third of known enzymes and other functional proteins such as RNA polymerase, DNA transcription factors, cytochromes P450s, hemoglobin, myoglobin, and coenzymes such as vitamin B.sub.12. There, these metals serve to facilitate oxidation and reduction reactions, stabilize or shield charge distributions, and orient substrates for reactions.

[0022] However, the body has a limited ability to absorb and excrete metals, and an excess can lead to toxicity. The principles underlying the present disclosure can be applied to any condition in which an excess of metals is the causative agent of a health condition.

[0023] As one example, an excess of iron, whether derived from red blood cells chronically transfused, necessary in such conditions such as β-thalassemia major, or from increased absorption of dietary iron such as hereditary hemochromatosis caused by mutation in genes such as HFE can be toxic through the generation by iron of reactive oxygen species such HO. and HOO. In the presence of Fe.sup.2+, H.sub.2O.sub.2 is reduced to the hydroxyl radical (HO.), a very reactive species, a process known as the Fenton reaction. The hydroxyl radical reacts very quickly with a variety of cellular constituents and can initiate free radicals and radical-mediated chain processes that damage DNA and membranes, as well as produce carcinogens. The clinical result is that without effective treatment, total body iron progressively increases with deposition in the liver, heart, pancreas, and elsewhere. Iron accumulation may also produce (i) liver disease that may progress to cirrhosis, (ii) diabetes related both to iron-induced decreases in pancreatic β-cell secretion and increases in hepatic insulin resistance and (iii) heart disease, still the leading cause of death in β-thalassemia major and other anemias associated with transfusional iron overload.

[0024] As another example, relative excess iron has been associated with increased risk of cardiovascular diseases (e.g., heart disease). There is a strong correlation between serum ferritin levels, inflammatory biomarkers such as C-reactive protein and interleukin-1, and mortality is a subset of patients with peripheral arterial disease; phlebotomy and iron chelation has been used to mitigate that risk. Treatment with an iron chelator is thought to reduce iron stores, reduce serum ferritin and potentially reduce the incidence of heart disease and stroke.

[0025] As another example, ions with little or no endogenous function may find their way into the body and effect damage. Heavy metal ions such as Hg.sup.2+ can replace ions such as Zn.sup.2+ in metalloproteins and render them inactive, resulting in serious acute or chronic toxicity that can end in a patient's death or in birth defects in that patient's children. Even more significantly, radioactive isotopes of the lanthanide and actinide series can visit grave illness on an individual exposed to them by mouth, air, or skin contact. Such exposure could result not only from the detonation of a nuclear bomb or a “dirty bomb” composed of nuclear waste, but also from the destruction of a nuclear power facility.

[0026] Any of the foregoing metal mediated conditions, as well as those identified in embodiments 46 and 89 to 105, can benefit from the cellular recovery dosing regimens disclosed herein. Accordingly, the present disclosure encompasses methods of treatment of such metal mediated conditions using the regimens described in Section 5.3 or identified using the assays identified in Section 5.4.

5.2 Metal Chelators

[0027] The cellular recovery dosing regimens of the disclosure pertain to metal chelating agents such as SP-420 compounds. As used herein, “SP-420 compounds” means SP-420 ((S)-4,5-dihydro-2-[2-hydroxy-4-(3,6-dioxaheptyloxy)phenyl]-4-methyl-4-thiazolecarboxylic acid) and pharmaceutically acceptable salts, solvates, and hydrates thereof.

[0028] Various agents for the chelation and decorporation of metal ions in living organisms are in clinical use and others have been disclosed and either are untested clinical or failed to advance in clinical trials due to toxicity. Any metal chelator based treatment can benefit reduced toxicity resulting from the cellular recovery regimens described in Section 5.3 or identified using the assays identified in Section 5.4.

[0029] Exemplary metal chelators to which the regimens apply include, but are not limited to, those identified in Sections 2 and 3, in embodiments 48, 70, 73, 76, 79, 86, and 106, and those compounds encompassed by Formulas (I)-(IX) described in PCT publication no. WO 2006/107626, the contents of which are incorporated in herein by reference thereto.

[0030] Some metal chelators of the disclosure, such as those described in the preceding paragraph, can be administered in the form of pharmaceutically acceptable salts. Pharmaceutically acceptable salts include alkali and alkaline earth metal (e.g., sodium, potassium, magnesium, calcium) salts. In addition, pharmaceutically acceptable salts include amine salts. Preferably, the pharmaceutically acceptable salts are metal-free (e.g., iron-free) salts. As used herein, “metal-free salt” does not encompass alkali and alkali earth metal salts.

5.3 Dosing Regimens

[0031] The cellular recovery dosing regimens of the disclosure are believed to have a net effect of increasing the therapeutic index of metal chelator therapy by preserving efficacy while minimizing toxicity.

[0032] The regimens generally reduce the dose frequency by increasing the period of time between doses (the dosing interval) to allow a period of time where chelator is not present in the cells of the body at sufficiently high concentrations to effectively chelate metal ions. This allows the cells to have functional metal containing proteins in sufficient amounts to have normal cellular function. However, the chelator dose and frequency are sufficient to create a state of metal balance or negative metal balance depending on the therapeutic goal of chelation therapy.

[0033] In the case of the chelators currently approved for therapy, in some embodiments the interval between doses is increased by approximately 30% to 300% (increasing the 24 hour interval for once per day dosing by approximately 4 hours to 72 hours). For example, a chelator such as deferasirox is typically given once per day on an empty stomach before the morning meal or with a light, low-fat breakfast. Thus the interval between doses is approximately 24 hours. In one scenario, the second dose would occur at the next day mid-day meal, extending the interval to approximately 28 hours. The third dose would be at the evening meal of the following day and so forth. In another scenario, the second dose would occur at the next day's evening meal, extending the interval to approximately 32 hours. In a third scenario, the second dose would occur 2 days later at the morning meal, extending the interval to approximately 48 hours. The predicted benefit of extending the interval between doses would be to allow the cells adversely affected by chelation of iron the necessary time to recover during an effectively chelator-free period. Although some chelator may be present, it would be at a sufficiently low concentration so as not to interfere with cellular function. In the case of deferasirox, this is predicted to result in substantially less kidney and gastrointestinal injury. More generally, it is predicted that dose-related toxicity observed with any chelator would be reduced or eliminated with the modified dosing regimen.

[0034] In certain aspects the dose administered with the new regimen would be increased to offset the reduced dosing frequency. The dose of chelator given at each administration would therefore be increased such that the intended amount of metal clearance was maintained with the new regimen. The dose can be increased by the addition of 10% to 300% of the original dose. By way of illustration, a 20 mg/kg dose of deferasirox is increased to a range of 22 to 60 mg/kg at each administration. All ranges described in the disclosure are inclusive (e.g., a range of 22 to 60 mg/kg encompasses a 22 mg/kg dose and a 60 mg/kg dose).

[0035] In some embodiments, the total weekly or monthly dose (the sum of all doses administered in a week or month) is maintained. For instance, if deferasirox was initially dosed at 20 mg/kg daily and the dosing interval was changed to every other day dosing, then the dose administered would increase to 40 mg/kg administered every other day.

[0036] In the dosing regimens described herein, the phrase “weekly doses” when preceded by an integer refers to the number of doses administered in a week. For example, a dosing regimen comprising “fewer than 5 weekly doses” encompasses dosing regimens in which the subject is administered a dose of a metal chelator fewer than five times in a week (e.g., 1, 2, 3, or 4 times in a week). Similarly, the phrase “daily doses” when preceded by an integer refers to the number of doses administered in a day. For example, a dosing regimen comprising “one or two daily doses” encompasses dosing regimens in which the subject is administered a dose of a metal chelator once per day or twice per day. When a specific dose of a metal chelator is provided, unless required otherwise by context, the dose refers to the amount of metal chelator administered in a single dose. The foregoing weekly, daily and specific doses can be administered for a period of at least one week, at least one month, at least three months, at least six months, at least one year, or indefinitely (e.g., for the remainder of a subject's life). Unless required otherwise by context, a period of administration refers to the amount of time between the first and last doses of a metal chelator, rather than the amount of time of a single administration.

[0037] The dosing regimen can be modified further to enhance convenience, compliance, and adherence. For instance, every other day dosing can be changed to dosing on three days per week, with, for example a weekend break. In geographies where the weekend is Saturday and Sunday, such a dosing schedule can consist of administering the chelator on Mondays, Wednesdays, and Fridays. It is believed that this would enhance the ability of a patient to remember to take the dose and thereby comply with the recommended regimen. Also, as shown in Example 2, a short break from dosing is expected to minimize chelator toxicity.

[0038] The metal chelating efficiency (the percentage of drug administered that is excreted from the body chelated with metal) can be determined with metal balance studies in animals, human clinical trial subjects, or patients or by measuring tissue metal content, for instance in animals using atomic absorption spectroscopy of tissue or in humans by MRI.

[0039] Since side-effects from chelators are either predictable or can be anticipated based on screening tests, the between dose interval can be adjusted over time while monitoring side-effects such as abdominal pain or examining the urine or blood for evidence of kidney injury or dysfunction.

[0040] For metal chelators without an established dose, the dosing schedule can be established using the principles described herein, for example using the assays described in Section 5.4.

[0041] In particular embodiments, the metal chelator is administered according to any of the regimens identified in embodiments 1 to 106 or by the methods described in Section 5.4.

[0042] The chelators used in the cellular recovery regimens of the disclosure are preferably in immediate release oral formulations but may also be delayed release oral dosage forms or in parenteral preparations. The cellular recovery regimens of the disclosure can be used for dosing humans (including both adults and children) and non-human animals.

[0043] 5.4 Assays for Optimizing Dosing Regimens

[0044] Various cell and animal models can be used to evaluate the optimal dosing intervals for metal chelators. These models probe the toxic effects on metal chelators on cells in sensitive organs to determine dose toxicity and cellular recovery time between doses.

[0045] The cell types studied in cell culture can be selected to reflect the cell type adversely affected in humans such as hepatocytes or granulocyte progenitor cells.

[0046] To evaluate the effects of metal chelators on renal toxicity, primary proximal tubule kidney cells or immortalized proximal tubule cell lines can be grown in tissue culture (in vitro) using defined culture media. Assays exist to quantify the metabolic status (e.g., MTTP assay), proliferation (e.g., cell counts), and vitality (e.g., trypan blue exclusion) of these cells. Metal (e.g., iron) chelators can be added to the culture medium for defined periods of time to model in vivo exposure of the kidney. For instance, following daily oral dosing of deferasirox at steady state in human patients, the peak serum concentration is achieved at approximately 2 hours after the dose and the mean elimination half-life of is approximately 12 hours. Deferasirox is highly bound to albumin so albumin would be added to the culture medium. To model this, the cells in culture can be exposed to the high peak serum concentration in the culture medium for 2 hours. The culture medium can be changed with fresh medium containing a lower concentration of deferasirox every 2 hours such that at the end of 14 hours (2 hours of initial high concentration plus 12 hours of declining concentration) the final concentration is approximately half that of the initial concentration. This every 2 hour reduction in deferasirox concentration is continued until the end of the 24 hour period resulting in a concentration that approximately equaled the trough concentration observed in patients taking deferasirox once per day chronically. This dosing scheme can be repeated for a number of days, for instance 4 days. To confirm that toxicity can be reduced using a cellular recovery regimen that incorporates a chelator free period in which the cells are able to recover metabolic functions, the cellular toxicity observed using an exposure schedule based on standard dosing of a metal chelator is then compared to the toxicity observed using an exposure schedule based on a cellular recovery dosing schedule. For example in the case of deferasirox, the cellular assay described above can be carried out using a reduced dosing schedule, for example involving exposing the cells to twice the concentrations of defersirox every other day for the same number of days to achieve the same overall dose.

[0047] In the case of other chelators, the assays are designed to compare a typical pharmacokinetic profile of that chelator with a modified profile that incorporates chelator free periods for cellular recovery. Deferiprone, for instance, is administered 3 times per day, typically once in the morning upon arising, once at mid-day, and once in the evening. Thus a typical regimen would result in doses separated by approximately 5 hours with a peak serum concentration achieved after 1 hour and a half-life of 2 hours. This can be mimicked by a high initial deferiprone concentration in vitro for 1 hour followed by a reduction in concentration of 50% for 2 hours and another reduction of 50% for a subsequent 2 hours. This cycle is repeated 2 more time to mimic the 3 times per day dosing in patients. To confirm that toxicity can be reduced using a cellular recovery regimen that incorporates a chelator free period in which the cells are able to recover metabolic functions, the cellular toxicity observed using an exposure schedule based on standard dosing of a metal chelator is then compared to a cellular recovery dosing schedule. In the case of deferiprone, the toxicity using an exposure schedule based on the standard dosing schedule is compared to an exposure schedule that results in the same overall dose administered in 2 divided doses at higher concentrations separated by 10 hours or as a single dose separated by 24 hours.

[0048] Kidney proximal tubule cells and other cells such as hepatocytes can also be cultured in microfluidic kidney models (“kidney-on-a-chip”) or liver models (“liver-on-a-chip”) and exposed to chelators in a manner that mimics the pharmacokinetic of various dosing regimens. In one embodiment, the “kidney-on-a-chip” model used to evaluate metal chelator toxicity and cellular recovery periods is that described by Kim and Takayama (Kim et al., 2015, Biofabrication. Volume 8, Number 1 015021, dx.doi.org/10.1088/1758-5090/8/1/015021). As with the simpler cell culture models, the metabolic status and vitality can be assessed in response to cellular recovery dosing regimens (e.g., every other day dosing of deferasirox or twice a day or once a day dosing for deferiprone) as compared to standard dosing regimens (e.g., daily dosing of deferasirox or thrice daily dosing for deferiprone).

[0049] The frequency of the dosing regimen can also be studied in animals. For instance, the same total dose of chelator (e.g., 30 mg/kg/day) can be given to one group of animals as 10 mg/kg three times per day, while another group of animals can be given 15 mg/kg twice per day, and a third group of animals can be given 30 mg/kg once per day. In an extension of the assay, another groups of animals can be given 60 mg/kg once every other day and still another group of animals can be given 90 mg/kg once every third day. These animals are monitored for overt toxicity and evidence of organ dysfunction (e.g., rise in serum creatinine or abnormal urinalysis or elevation of liver enzymes), or changes in tissues (e.g., vacuolization of kidney proximal tubule cells or accumulation of fat in hepatocytes).

[0050] Information obtained from these assays can be used to guide dose selection for human clinical trials to compare standard chelator dosing regimens with modified regimens that incorporate periods of cellular recovery from chelator therapy. The modified regimens can also be studied alone and compared to historical control data, e.g., from trials with standard regimens.

6. EXAMPLES

6.1 Example 1: Comparison of Daily Versus Every Other Day Dosing of SP-420

[0051] This study was performed to compare the toxicity of daily versus every other day dosing of SP-420 (administered as the sodium salt) by assessing levels of kidney injury molecule-1 (KIM-1) and the presence of glucose in the urine (glucosuria). KIM-1 is a sensitive and specific urinary biomarker of kidney injury in both rodent models and humans (see, e.g., Sabbisetti et al., 2014, J Am Soc Nephrol. 25(10):2177-2186). KIM-1 is minimally expressed in normal rat kidneys, but is markedly expressed upon kidney injury (see, e.g, Vaidya et al., 2010, Nat Biotechnol. 28(5): 478-485). Glucose filtered by the kidney glomerulus is normally completely reabsorbed by kidney proximal tubule cells and therefore it is very abnormal to detect glucose in the urine. Glucosuria is a condition where excessively large amounts of glucose are filtered the glomerulus, overwhelming the absorptive mechanism. Glucosuria is generally only detected in diabetes mellitus secondary to hyperglycemia or when the kidney proximal tubule cells become dysfunctional, as in the renal Fanconi syndrome. The renal Fanconi syndrome has been reported to be caused by iron chelator treatment. Resorption of glucose and other filtered substances (e.g., amino acids, phosphorus, uric acid) is highly energy dependant and chelation of kidney proximal tubule cell iron results in decreased mitochondrial energy production leading to cellular dysfunction with decreased resorptive capability.

6.1.1. Methods

[0052] Male Sprague-Dawley rats (n=5 per group) were given SP-420 orally by gavage once daily (SID) at a dose of 162.5 mg/kg or every other day (EOD) at a dose of 325 mg/kg. Each group of rats received a cumulative dose of 2600 mg/kg of SP-420. Urine was collected from metabolic cages EOD during the course of administration, 5 or 6 days after the last dose, and again 11 or 12 days after the last dose, and assessed for its KIM-1 content and subjected to a 10-parameter urine dipstick analysis. The urine from the animals administered SP-420 EOD was collected during the 24 hours immediately following the dose of the drug.

6.1.2. Results

[0053] All of the SID and EOD-treated rats survived the exposure to the drug. However, one rat in the 162.5 mg/kg SID group was euthanized 3 days post-drug due to ongoing weight loss and deteriorating condition. Urinary KIM-1 levels increased in both groups, but the increase was not as significant in animals given the drug EOD. Changes in the KIM-1 levels largely returned to baseline levels by 5 or 6 days post-drug. Glucosuria was observed in 4/5 of the rats administered SP-420 SID, but in none of the rats that were administered SP-420 EOD.

[0054] This study shows that administering SP-420 every other day rather than every day reduces toxicity.

6.2 Example 2: Repeat Study of Example 1

[0055] In Example 1, “recovery” KIM-1 levels were not measured until 5 or 6 days post last dose. Although it was clear that the KIM-1 levels dropped dramatically during this time period, the study in Example 1 was not designed to show how quickly the parameters returned to baseline levels. In addition, in Example 1 urine was collected from the rats given the chelator EOD in the 24 hours immediately following the dose of the drug but no data were collected for urine produced in the 24-48 hour time period leading up to the next dose of SP-420. In order to determine what occurred during this “off” day and to determine whether urinary KIM-1 levels decrease between doses, the study of Example 1 was repeated with slight modifications.

6.2.1. Methods

[0056] Ten male Sprague-Dawley rats were randomly divided into two groups of five and dosed as in Example 1. Unlike in Example 1, in which the rats were housed in the metabolic cages EOD, the animals in this repeat study were housed individually in metabolic cages throughout the course of the dosing regimen, and for 6 days thereafter to allow for the collection of urine during a recovery period. Urine was collected at 24-h intervals throughout the course of the study and assessed for its KIM-1 content. For the rats receiving the drug EOD, KIM-1 data was obtained both 0-24 hours immediately following the dose of the drug, as well as from the “off” day (24-48 hours post drug). The rats were weighed once daily. Changes in body weight were calculated from the animals' baseline weight versus their weight on the day that the last dose of the drug was given.

6.2.2. Results

[0057] All of the rats survived the exposure to the test drug. One rat from the 162.5 mg/kg SID group was euthanized 4 days post-last dose due to weight loss and overall deteriorating condition.

[0058] Representative KIM-1 data for a rat in each group is shown in FIG. 1A-1B. Although FIG. 1A-1B show that SP-420 induced increased KIM-1 levels in both groups of animals, the values returned to near baseline levels very quickly. The increase in KIM-1 levels in rats administered SP-420 EOD was not as significant in animals administered SP-420 SID. This is depicted in the KIM-1 strips shown in FIG. 2. Strips numbered 1-5 were from rats administered SP-420 SID at 162.5 mg/kg/day, while strips numbered 6-10 were from rats administered SP-420 at 325 mg/kg EOD. The “C” line is an internal standard, while the “T” line represents the results for the test animal. The “T” line in strips 3-5 are quite pronounced, while very little KIM-1 was observed in any of strips 6-10 except strip 7. KIM-1 levels decreased from the last administration to 5-days post-administration, as shown in FIG. 3. FIG. 3 shows KIM-1 strips from the rats administered SP-420 SID at the end of the administration period (left side) and 5 days post-administration (right side). Very little KIM-1 was observed in strip #4 or #5 five days post-drug. The rat corresponding to strip #3 was euthanized prior to obtaining the 5 days post-administration sample.

[0059] Under the conditions of the study, KIM-1 levels did not decrease in rats treated EOD on the “off” day (as shown in representative FIG. 1B). This was also observed in the recovery data of the rats treated with SP-420 SID, where the KIM-1 levels were virtually unchanged between what was seen on the last day of dosing versus the quantity of KIM-1 in the urine 24 hours post drug. However, the lack of a decrease in KIM-1 between doses in the EOD animals, or within 24 hours of stopping the drug in the SID rodents, is not surprising. The rats in these studies generally excrete less than 10-15 mL of urine per day. Due to such a small quantity of urine being produced, it is quite likely that there was significant carry over of KIM-1, glucose, etc., from one day to the next. For example, the picture shown in FIG. 4 depicts the 24-hour urine sample of a rat treated SID with SP-420 at a dose of 200 mg/kg/day. The volume of urine produced on that day was only 14.6 mL. While the bottom half of the tube contained normal looking urine, the top half contained a layer of frank blood. Based on the “layering effect” of normal urine versus urine containing blood observed during this single 24-hour sample, it is not unreasonable to expect that some of the KIM-1 or glucose present at the end of one 24-hour collection period would continue to be excreted into the following day's urine specimen as well.

[0060] Average KIM-1 data for all rats in each group is shown in FIG. 5. Although FIG. 5 shows that SP-420 induced increased average KIM-1 levels in both groups of animals, the average values returned to near baseline levels very quickly. The increase in average KIM-1 levels in rats administered SP-420 EOD was not as significant in animals administered SP-420 SID.

[0061] Glucosuria was observed in 3/5 rats given SP-420 SID. During the recovery period, glucosouria continued to be observed in all three of the rats one day post-drug, in 1/3 of the rats 2 days post-drug, and in none of the affected rats by 3 days post-last dose. Glucosuria was not observed in any of the rats administered SP-420 EOD at 325 mg/kg.

6.2.3. Summary

[0062] Example 2 shows that administering SP-420 every other day versus once daily reduces toxicity. This is reflected both in terms of urinary KIM-1 excretion, as well as the presence/absence of glucose in the test animals' urine. The rapid decrease in urinary KIM-1 levels on days 2-6 post drug provides strong support for the idea of administering the chelator to patients on a three days a week dosing schedule with a weekend break, e.g., a Monday, Wednesday, Friday dosing schedule. With a Monday, Wednesday and Friday dosing schedule the weekend break would, if necessary, allow the kidneys time to recover before dosing resumed on Monday.

6.2.4. Example 3: Comparison of Daily Versus Every Other Day Dosing of SP-420 at Different Doses

[0063] Studies similar to those described in Examples 1 and 2 were performed with 250 mg/kg EOD, 250 mg/kg SID, 500 mg/kg EOD, 200 mg/kg SID and 400 mg/kg EOD of SP-420. The results of the studies, together with the results of the studies described in Examples 1 and 2, are summarized in Table 1, below. As shown in Table 1, glucosuria and KIM-1 levels were consistenly lower for EOD dosing regimens compared to SID regimens for the same cumulative dose. Taken together, the data support for the idea of administering SP-420 to patients on an every other day or a three times a week (e.g., Monday, Wednesday, Friday) dosing schedule.

TABLE-US-00001 TABLE 1 Tolerability of SP-420 given Orally by Gavage to Male Sprague-Dawley Rats (Sodium Salt) Dose & Frequency 250 mg/kg 250 mg/kg 500 mg/kg 162.5 mg/kg 325 mg/kg 200 mg/kg 400 mg/kg Control EOD SID EOD SID EOD SID EOD Number of Rats 5 5 5 5 10 10 5 5 per Group Maximum Cumulative 0 2500   2500   2500   2600  2600  2400   2400   Dose (mg/kg) Survival 5/5 5/5 2/5 0/5 8/10 10/10  4/5 3/5 Deaths @ All All 2 @ 2000 1 @ 1500 2 @ 2600 All 1 @ 2000 1 @ 2000 Cumulative Survived Survived 1 @ 2500 2 @ 2000 1-3 d post Survived 1 @ 2400 Dose (mg/kg) 2 @ 2500 1-4 d post Weight +16.0 ± +8.5 ± −5.2 ± −5.2 ± −1.1 ± +0.8 ± −4.9 ± −2.8 ± Change (%) 1.4 3.9 4.0 5.0 5.0 2.5 5.3 5.6 Weight +13.7 to +3.1 to −9.1 to −13.1 to −7.7 to −3.0 to −8.3 to −8.8 to Change (Range, %) +17.0 +12.4 +0 −1.2 +5.5 +3.4 +4.4 +2.9 Urine Dipstick Glucosuria 0/5 0/5 4/5 1/5 7/10 0/10 3/5 1/5 Hematuria (Frank) 0/5 0/5 1/5 1/5 0/10 0/10 2/5 0/5 pH ≤5 0/5 0/5 3/5 2/5 5/10 0/10 4/5 2/5 Specific gravity ≥1.030 0/5 0/5 2/5 1/5 4/10 2/10 3/5 1/5 Kidney Injury Molecule (KIM-1) KIM-1 ≥250 ng/kg/24 h 0/5 0/5 5/5 4/5 8/10 9/10 5/5 4/5 KIM-1 ≥500 ng/kg/24 h 0/5 0/5 5/5 2/5 7/10 4/10 5/5 2/5 KIM-1 ≥1000 ng/kg/24 h 0/5 0/5 3/5 1/5 7/10 3/10 5/5 2/5 KIM-1 ≥2000 ng/kg/24 h 0/5 0/5 0/5 0/5 4/10 1/10 1/5 1/5

7. SPECIFIC EMBODIMENTS, CITATION OF REFERENCES

[0064] While various specific embodiments have been illustrated and described, it will be appreciated that various changes can be made without departing from the spirit and scope of the disclosure(s). The present disclosure is exemplified by the numbered embodiments set forth below. [0065] 1. A method of (a) treating a metal-mediated condition and/or (b) reducing the renal, gastrointestinal, hepatic, hematological, auditory, ocular, and/or skin toxicity of a metal chelator in a subject, comprising orally administering to the subject a plurality of doses of a metal chelator, each dose given more than 24 hours after the prior dose and/or no more than five times a week. [0066] 2. A method of (a) treating a metal-mediated condition and/or (b) reducing the renal, ocular, auditory, neurological, respiratory, and/or musculoskeletal toxicity of a metal chelator in a subject, comprising intravenously or subcutaneously administering to the subject fewer than 5 weekly doses of a metal chelator. [0067] 3. The method of embodiment 1, wherein the metal chelator is administered five times per week. [0068] 4. The method of embodiment 1 or embodiment 2, wherein the metal chelator is administered four times per week. [0069] 5. The method of embodiment 1 or embodiment 2, wherein the metal chelator is administered three times per week. [0070] 6. The method of embodiment 1, wherein the metal chelator is administered 15-25 times per month. [0071] 7. The method of embodiment 1, wherein the metal chelator is administered 20-25 times per month. [0072] 8. The method of embodiment 1 or embodiment 2, wherein the metal chelator is administered 12-18 times per month. [0073] 9. The method of embodiment 1 or embodiment 2, wherein the metal chelator is administered 15-20 times per month. [0074] 10. The method of any one of embodiments 1 to 9, wherein the metal chelator is administered every 28 to 72 hours. [0075] 11. The method of embodiment 10, wherein the metal chelator is administered every 28 hours. [0076] 12. The method of embodiment 10, wherein the metal chelator is administered every 32 hours. [0077] 13. The method of embodiment 10, wherein the metal chelator is administered every 36 hours. [0078] 14. The method of embodiment 10, wherein the metal chelator is administered every 42 hours. [0079] 15. The method of embodiment 10, wherein the metal chelator is administered every 48 hours. [0080] 16. The method of embodiment 10, wherein the metal chelator is administered every two days. [0081] 17. The method of embodiment 10, wherein the metal chelator is administered every 60 hours. [0082] 18. The method of embodiment 10, wherein the metal chelator is administered every 72 hours. [0083] 19. The method of embodiment 10, wherein the metal chelator is administered every three days. [0084] 20. The method of embodiment 10, wherein the metal chelator is administered every 28 to 36 hours. [0085] 21. The method of embodiment 10, wherein the metal chelator is administered every 32 to 48 hours. [0086] 22. The method of embodiment 10, wherein the metal chelator is administered every 36 to 48 hours. [0087] 23. The method of embodiment 10, wherein the metal chelator is administered every 32 to 60 hours. [0088] 24. The method of embodiment 10, wherein the metal chelator is administered every 36 to 60 hours. [0089] 25. The method of embodiment 10, wherein the metal chelator is administered every 48 to 60 hours. [0090] 26. The method of embodiment 10, wherein the metal chelator is administered every 32 to 72 hours. [0091] 27. The method of embodiment 10, wherein the metal chelator is administered every 36 to 72 hours. [0092] 28. The method of embodiment 10, wherein the metal chelator is administered every 48 to 72 hours. [0093] 29. A method of (a) treating a metal-mediated condition and/or (b) reducing the hematological, gastrointenstinal, hepatic and/or musculoskeletal toxicity of a metal chelator in a subject, wherein the method comprises orally administering to the subject one or two daily doses of the metal chelator, and wherein the metal chelator is deferiprone or a pharmaceutically acceptable salt, solvate or hydrate thereof. [0094] 30. The method of embodiment 29, which comprises administering two daily doses of the metal chelator. [0095] 31. The method in embodiment 29, wherein the daily doses are administered 6, 9, or 12 hours apart. [0096] 32. The method of embodiment 29, wherein the metal chelator is administered fourteen times per week. [0097] 33. The method of any one of embodiments 1 to 32, wherein the metal chelator is administered for a period of at least one week. [0098] 34. The method of any one of embodiments 1 to 32, wherein the metal chelator is administered for a period of at least one month. [0099] 35. The method of any one of embodiments 1 to 32, wherein the metal chelator is administered for a period of at least three months. [0100] 36. The method of any one of embodiments 1 to 32, wherein the metal chelator is administered for a period of at least six months. [0101] 37. The method of any one of embodiments 1 to 32, wherein the metal chelator is administered for a period of at least one year. [0102] 38. The method of any one of embodiments 1 to 32, wherein the metal chelator is administered indefinitely. [0103] 39. The method of any one of embodiments 1 to 38, wherein a dose that is greater than the standard daily dose is given at each administration. [0104] 40. The method of any one of embodiments 1 to 39, wherein the aggregate weekly dose administered is equal to the standard aggregate weekly dose. [0105] 41. The method of any one of embodiments 1 to 39, wherein the aggregate weekly dose administered is less than the standard aggregate weekly dose. [0106] 42. The method of embodiment 41, wherein the aggregate weekly dose administered is 0.75 to 0.9 times the standard aggregate weekly dose. [0107] 43. The method of any one of embodiments 1 to 38, wherein the standard daily dose is given at each administration and/or the aggregate weekly dose administered is equal to the standard aggregate weekly dose. [0108] 44. The method of any one of embodiments 1 to 39, wherein the aggregate weekly dose administered is greater than the standard aggregate weekly dose. [0109] 45. The method of embodiment 44, wherein the aggregate weekly dose administered is 1.25 to 2 times the standard aggregate weekly dose. [0110] 46. The method of any one of embodiments 1 to 45, wherein the metal is iron. [0111] 47. The method of embodiment 46, wherein the metal chelator is an iron chelator. [0112] 48. The method of any one of embodiments 1 to 47, wherein the metal chelator is other than an SP-420 compound. [0113] 49. The method of embodiment 47 except when dependent directly or indirectly from any one of embodiments 29 to 32 or 39 to 45, wherein the iron chelator is an SP-420 compound. [0114] 50. The method of embodiment 49, wherein the SP-420 compound is administered at a dose of 18-100 mg/kg. [0115] 51. The method of embodiment 49, wherein the SP-420 compound is administered at a dose of 18-30 mg/kg. [0116] 52. The method of embodiment 49, wherein the SP-420 compound is administered at a dose of 24 mg/kg. [0117] 53. The method of embodiment 49, wherein the SP-420 compound is administered at a dose of 40-60 mg/kg. [0118] 54. The method of embodiment 49, wherein the SP-420 compound is administered at a dose of 60-84 mg/kg. [0119] 55. The method of embodiment 49, wherein the SP-420 compound is administered at a dose of 72 mg/kg. [0120] 56. The method of embodiment 49, wherein the SP-420 compound is administered at a dose ranging between 500 mg and 10 g. [0121] 57. The method of embodiment 49, wherein the SP-420 compound is administered at a dose ranging between 1 g and 5 g. [0122] 58. The method of any one of embodiments 49 to 57, wherein the SP-420 compound is administered at a total weekly dose of 54-400 mg/kg. [0123] 59. The method of embodiment 58, wherein the SP-420 compound is administered at a total weekly dose of 54-100 mg/kg. [0124] 60. The method of embodiment 58, wherein the SP-420 compound is administered at a total weekly dose of 100-200 mg/kg. [0125] 61. The method of embodiment 58, wherein the SP-420 compound is administered at a total weekly dose of 200-300 mg/kg. [0126] 62. The method of embodiment 58, wherein the SP-420 compound is administered at a total weekly dose of 300-400 mg/kg. [0127] 63. The method of any one of embodiments 49 to 57, wherein the SP-420 compound is administered at a total weekly dose ranging between 1.5 g and 30 g. [0128] 64. The method of embodiment 63, wherein the SP-420 compound is administered at a total weekly dose ranging between 3 g and 30 g. [0129] 65. The method of embodiment 63, wherein the SP-420 compound is administered at a total weekly dose ranging between 5 g and 20 g. [0130] 66. The method of any one of claims 49 to 65, wherein the SP-420 compound is SP-420 ((S)-4,5-dihydro-2-[2-hydroxy-4-(3,6-dioxaheptyloxy)phenyl]-4-methyl-4-thiazolecarboxylic acid). [0131] 67. The method of any one of embodiments 49 to 65, wherein the SP-420 compound is a pharmaceutically acceptable salt of SP-420. [0132] 68. The method of any one of embodiments 49 to 65, wherein the SP-420 compound is a pharmaceutically acceptable solvate of SP-420. [0133] 69. The method of any one of embodiments 49 to 65, wherein the SP-420 compound is a pharmaceutically acceptable hydrate of SP-420. [0134] 70. The method of embodiment 47, wherein the iron chelator is deferiprone or a pharmaceutically acceptable salt, solvate or hydrate thereof. [0135] 71. The method of embodiment 70, wherein the standard dose of deferiprone is 25 to 33 mg/kg three times per day by oral administration. [0136] 72. The method of embodiment 70, wherein the dose of deferiprone administered to the subject is 37.5 to 49.5 mg/kg twice per day or 66 to 99 mg/kg once per day. [0137] 73. The method of embodiment 47 except when dependent directly or indirectly from any one of embodiments 29 to 33, wherein the iron chelator is deferoxamine or a pharmaceutically acceptable salt, solvate or hydrate thereof. [0138] 74. The method of embodiment 73, wherein the standard dose of deferoxamine is 20 to 50 mg/kg administered as a slow subcutaneous or intravenous infusion over 8-12 hours 5 to 7 days per week or 500 to 1000 mg administered as a daily injection. [0139] 75. The method of embodiment 73, wherein the dose of deroxamine administered to the subject is 20 to 50 mg/kg administered as a slow subcutaneous over 4-6 hours 5 to 7 days per week or as 40 to 100 mg/kg administered as a slow subcutaneous or intravenous infusion over 8 to 12 hours 2 to 3 days per week or as 1000 to 2000 mg administered as an every other day injection. [0140] 76. The method of embodiment 47 except when dependent directly or indirectly from any one of embodiments 29 to 33, wherein the iron chelator is deferasirox or a pharmaceutically acceptable salt, solvate or hydrate thereof. [0141] 77. The method of embodiment 76, wherein the standard dose of deferasirox is 20-40 mg/kg administered orally once per day. [0142] 78. The method of embodiment 76, wherein the dose of deferasirox administered to the subject is 40-80 mg/kg administered orally once every other day. [0143] 79. The method of embodiment 47 except when dependent directly or indirectly from any one of embodiments 29 to 33, wherein the iron chelator is deferitrin or a pharmaceutically acceptable salt, solvate or hydrate thereof. [0144] 80. The method of embodiment 79, wherein the standard dose of deferitrin is 10 to 80 mg/kg administered orally once per day. [0145] 81. The method of embodiment 79, wherein the standard dose of deferitrin is 10 to 60 mg/kg administered orally once per day. [0146] 82. The method of embodiment 79, wherein the standard dose of deferitrin is 10 to 40 mg/kg administered orally once per day. [0147] 83. The method of embodiment 79, wherein the dose of deferitrin administered to the subject is 20 to 160 mg/kg administered orally once every other day. [0148] 84. The method of embodiment 79, wherein the dose of deferitrin administered to the subject is 20 to 120 mg/kg administered orally once every other day. [0149] 85. The method of embodiment 79, wherein the dose of deferitrin administered to the subject is 20 to 80 mg/kg administered orally once every other day. [0150] 86. The method of embodiment 47 except when dependent directly or indirectly from any one of embodiments 29 to 33, wherein the iron chelator is SPD602 (FBS0701) or a pharmaceutically acceptable salt, solvate or hydrate thereof. [0151] 87. The method of embodiment 86, wherein the standard dose of SPD602 is 10 to 60 mg/kg administered orally once per day. [0152] 88. The method of embodiment 86, wherein the dose of SPD602 administered to the subject is 20 to 120 mg/kg administered orally once every other day. [0153] 89. The method of any one of embodiments 46 to 88, wherein the metal-mediated condition is iron overload. [0154] 90. The method of embodiment 89, wherein the metal-mediated condition is the result of mal-distribution or redistribution of iron in the body. [0155] 91. The method of embodiment 89, wherein the metal-mediated condition is atransferrinemia, aceruloplasminemia, or Fredreich's ataxia. [0156] 92. The method of embodiment 89, wherein the metal-mediated condition is the result of transfusional iron overload. [0157] 93. The method of embodiment 89, wherein the metal-mediated condition is β-thalassemia. [0158] 94. The method of embodiment 93, wherein the metal-mediated condition is β-thalassemia major. [0159] 95. The method of embodiment 93, wherein the metal-mediated condition is β-thalassemia intermedia. [0160] 96. The method of embodiment 89, wherein the metal-mediated condition is sickle cell anemia, Diamond-Blackfan anemia, sideroblastic anemia, chronic hemolytic anemia, off-therapy leukemia, bone marrow transplant or myelodysplastic syndrome. [0161] 97. The method of embodiment 89, wherein the metal-mediated condition is a hereditary condition resulting in the excess absorption of dietary iron. [0162] 98. The method of embodiment 97, wherein the metal-mediated condition is hereditary hemochromatosis or porphyria cutanea tarda. [0163] 99. The method of embodiment 89, wherein the metal-mediated condition is an acquired disease that results in excess dietary iron absorption. [0164] 100. The method of embodiment 99, wherein the metal-mediated condition is a liver disease. [0165] 101. The method of embodiment 100, wherein the liver disease is hepatitis. [0166] 102. The method of any one of embodiments 1 to 28 and 33 to 45 except when embodiments 33 to 45 are dependent directly or indirectly from any one of embodiments 29 to 33, wherein the metal-mediated condition is lanthanide or actinide overload. [0167] 103. The method of embodiment 102, wherein the metal chelator is diethylene triamine pentaacetic acid (DTPA). [0168] 104. The method of any one of embodiments 1 to 28 and 33 to 45 except when embodiments 33 to 45 are dependent directly or indirectly from any one of embodiments 29 to 33, wherein the metal is lead or mercury. [0169] 105. The method of embodiment 104, wherein the metal-mediated condition is lead or mercury poisoning. [0170] 106. The method of embodiment 105, wherein the metal chelator is edetate calcium disodium or ethylenediaminetetraacetic acid (EDTA).

[0171] All publications, patents, patent applications and other documents cited in this application are hereby incorporated by reference in their entireties for all purposes to the same extent as if each individual publication, patent, patent application or other document were individually indicated to be incorporated by reference for all purposes. In the event that there is an inconsistency between the teachings of one or more of the references incorporated herein and the present disclosure, the teachings of the present specification are intended.