Neuroblastoma treatment with taurolidine hydrolysis products

Abstract

Neuroblastoma is a tumor primarily affecting children. The current standard of care is not curative except in the rare case of a surgically-resectable lesion, although very high survival rates have been documented for low-risk neuroblastoma and moderate-risk neuroblastoma. Taurolidine was developed as an anti-infective, but it has been found to have surprising oncolytic activity in cell cultures and now in a rodent cancer model. The efficacy in rodent model is superior to the efficacy in cell culture. This invention relates to the use of taurolidine hydrolysis products (tarultam and/or taurinamide and/or methylene glycol and/or selected combinations thereof) for the treatment of neuroblastoma in juvenile mammals.

Claims

1. A method for treating neuroblastoma, the method comprising: administering a composition to a patient, wherein the composition comprises: taurultam and taurinamide in a weight ratio of 1 taurultam:7 taurinamide, with a dosage range of taurultam between 5 mg/kg and 40 mg/kg and a dosage range of taurinamide from 35 mg/kg to 280 mg/kg; wherein the composition is pegylated using polyethylene glycols (PEGs) to delay premature hydrolysis of the composition; and wherein the composition is administered from once daily through weekly, for an effective period of time based on the patient's response.

2. The method according to claim 1 wherein the composition is administered in conjunction with an oncolytic agent and/or radiotherapy.

3. The method according to claim 1 wherein the composition is delivered to the patient using one from the group consisting of parenteral delivery, intramuscular delivery and intravenous delivery.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) These and other objects and features of the present invention will be more fully disclosed or rendered obvious by the following detailed description of the preferred embodiments of the invention, which is to be considered together with the accompanying drawings wherein like numbers refer to like parts, and further wherein:

(2) FIG. 1 is a graph showing that leukemia cell lines appear more sensitive to the effects of taurolidine compared to healthy lymphocytes in vitro (not in vivo);

(3) FIG. 2 is a graph showing that neuroblastoma cell lines are more sensitive to a decrease in viability due to taurolidine when compared to healthy fibroblasts (BJ on graph) in vitro (not in vivo);

(4) FIGS. 3-6 are graphs or photographs showing that taurolidine given to CB57 SCID mice with measurable tumors from a neuroblastoma cell line implanted subcutaneously in the CB57 SCID mice has efficacy in IMR5 tumors and measurable efficacy in SK-N-AS tumors in vivo (not in vitro);

(5) FIGS. 7 and 8 are graphs showing that statistically significant decreases in tumor size were achieved when taurolidine was administered to treat mice with a different cell line (SK-N-AS) also derived from neuroblastoma but overall survival was not significantly different from control;

(6) FIG. 9 illustrates the mechanism for the hydrolysis of taurolidine;

(7) FIG. 10 is a chart showing the mean pharmacokinetic parameters of taurinamide; and

(8) FIG. 11 is a chart showing the mean pharmacokinetic parameters of taurultam.

DETAILED DESCRIPTION OF THE INVENTION

(9) Taurolidine is a well known antimicrobial with a published mechanism of action and antimicrobial spectrum. Taurolidine is unstable in circulation and therefore has not been successfully developed for systemic infections. Taurolidine has demonstrated efficacy in local application for peritonitis and for prevention of infection when infused as a catheter-lock solution.

(10) Taurolidine has recently been investigated for oncolytic activity and found to have inhibitory effect on cell lines in culture, in combination with standard chemotherapy or alone. Despite claims that in vitro inhibitory concentrations are clinically achievable, the only published human pharmacokinetic study showed NO measurable concentration of taurolidine in healthy volunteers when 5 grams of taurolidine were given intravenously by 20 minute infusion. This is believed to be due to the rapid hydrolysis of taurolidine when administered systemically in a mammalian body.

(11) It has been found that leukemia cell lines appear more sensitive to the effects of taurolidine compared to healthy lymphocytes in vitro (not in vivo). See FIG. 1.

(12) It has also been found that neuroblastoma cell lines are more sensitive to a decrease in viability due to taurolidine when compared to healthy fibroblasts in vitro (not in vivo). See FIG. 2.

(13) Furthermore, taurolidine given to CB57 SCID mice with measurable tumors from a neuroblastoma cell line implanted subcutaneously in the CB57 SCID mice showed efficacy in IMR5 tumors and measurable efficacy in SK-N-AS tumors in vivo (not in vitro). See FIGS. 3-6.

(14) Note that the in vitro efficacy for neuroblastoma cell lines is seen at the highest two concentrations tested, i.e., above 40 microMolar [1 Mole/Liter×284 gm/Mole×1 Mole/1,000,000 microMoles×40 microMolar×1000 mg/gram=11 mg/liter=11 mcg/mL], as seen in FIG. 2 (where only the SK-N-MC cell line, a neuroepithelioma cell line, is below 50% cell viability). The IC50 values are between 80-140 microMolar for neuroblastoma cell lines, and 200 microMolar for normal fibroblasts (see FIG. 2).

(15) The efficacy observed with taurolidine treatment of IMR5 cell implants in vivo (FIGS. 3-6), despite IRM5's relatively high IC50 in vitro (FIG. 2), supports the conclusion that the hydrolysis products of taurolidine may have independent anti-neoplastic activity. In other words, taurolidine treatment of neuroblastoma cells in vivo appears to be significantly more effective than taurolidine treatment of neuroblastoma cells in vitro. Since it is known that taurolidine metabolizes to taurolidine hydrolysis products in vivo, this would support the conclusion that the hydrolysis products of taurolidine may have significant anti-neoplastic activity against neuroblastoma cells. In fact, it may be that the hydrolysis products of taurolidine have higher efficacy against neuroblastoma cells than taurolidine which has not been hydrolyzed.

(16) Statistically significant decreases in tumor size were achieved when taurolidine was administered to treat mice with a different cell line (SK-N-AS) also derived from neuroblastoma, though overall survival of the mice implanted with the tumor was not statistically different from the control. See FIGS. 7 and 8.

(17) It has now been discovered that selected hydrolysis products of taurolidine may be used to treat neuroblastoma. The mechanism for the hydrolysis of taurolidine is shown in FIG. 9. The selected hydrolysis products of taurolidine that may be used to treat neuroblastoma may comprise at least one from the group consisting of:

(18) taurinamide;

(19) taurultum;

(20) methylene glycol;

(21) taurultam and taurinamide in a ratio of 1 taurultam:7 taurinamide; and

(22) taurultam, taurinamide and methylene glycol in a ratio of 1 taurultam:7 taurinamide:1 methylene glycol.

(23) The taurinamide is given with a dosage range of from 5 mg/kg to 280 mg/kg, with optimal range between 5 mg/kg and 60 mg/kg, from once daily through weekly, for an effective period of time based on individual patient response. The mean pharmacokinetic parameters of taurinamide are shown in FIG. 10.

(24) The taurultam is given with a dosage range of from 5 mg/kg to 280 mg/kg, with optimal range between 5 mg/kg and 60 mg/kg, from once daily through weekly, for an effective period of time based on individual patient response. The mean pharmacokinetic parameters of taurultam are shown in FIG. 11.

(25) The methylene glycol is given with a dosage range of from 2.5 mg/kg to 160 mg/kg, with optimal range between 2.5 mg/kg and 30 mg/kg, from once daily through weekly, for an effective period of time based on individual patient response.

(26) The taurultam and taurinamide (in a ratio of 1 taurultam:7 taurinamide) is given with a dosage range of Taurultam from 5 mg/kg to 280 mg/kg, with optimal range between 5 mg/kg and 40 mg/kg, combined with Taurinamide from 5 mg/kg to 280 mg/kg, with optimal range from 35 mg/kg to 40 mg/kg, from once daily through weekly, for an effective period of time based on individual patient response.

(27) The taurultam, taurinamide and methylene glycol (in a ratio of 1 taurultam:7 taurinamide:1 methylene glycol) is given with a dosage range of Taurultam from 5 mg/kg to 280 mg/kg, with optimal range between 5 mg/kg and 40 mg/kg, combined with Taurinamide with a dosage range of from 5 mg/kg to 280 mg/kg, with optimal range from 35 mg/kg to 40 mg/kg, further combined with methylene glycol with a dosage range from 2.5 mg/kg to 160 mg/kg, with optimal range from 5 mg/kg to 40 mg/kg, from once daily through weekly, for an effective period of time based on individual patient response.

(28) Dose selection for the hydrolysis products were calculated as follows:
AUC 0-inf Taurultam/AUC 0-inf Taurinamide=42.9/312.7=0.14

(29) Since the molecular weight difference is only a single methyl group, the use of weight-based AUC does not need to be corrected. Therefore, the target ratio when giving Taurultam and Taurinamide in combination is 0.14 or 1:7. And the target ratio when giving taurultam and taurinamide and methylene glycol in combination is 1:7:1.

(30) Effective dosage was computed by computing the human equivalent dosage from the effective mouse dose using the formula:
[Human equivalent dose=mouse mg/kg dose×1 adult human/12 mice×25 child BSA ratio/37 adult BSA ratio=child dose in mg/kg
(https://www.fda.gov/downloads/drugs/guidances/ucm0789 32.pdf).

(31) In one preferred form of the invention, the selected hydrolysis products (active ingredient) can be delivered systemically in a “shielded form” so that they can reach the site of the neuroblastoma to avoid premature degradation.

(32) More particularly, in one preferred form of the invention, the hydrolysis products can be delivered in the form of a nanoparticle, where the nanoparticle comprises a core of the hydrolysis product and an exterior coating which is configured to prevent premature exposure of the hydrolysis product prior to the arrival of the nanoparticle to the tumor site. The exterior coating breaks down as the nanoparticle travels from the site of insertion to the site of the tumor so as to release the hydrolysis product intact at the site of the tumor. In one preferred form of the invention, the coating comprises an absorbable polymer or lipid which breaks down as the nanoparticle travels from the site of insertion to the site of the tumor. By way of example but not limitation, the coating can be created from combinations of copolymers and multimers derived from polymers structured from 1-lactide, glycolide, e-caprolactone, p-dioxanone, and trimethylene carbonate. The coating may also be associated with glycols such as polyethylene glycols (PEGs), which can either be linear or multi-arm structures.

(33) If desired, the nanoparticle may comprise an excipient (e.g., a buffer for providing enhanced hydrolytic stability of the hydrolysis product within the nanoparticle).

(34) Additionally, if desired, the nanoparticle can further comprise a coating, wherein the coating is configured to target the nanoparticle to the site of a neuroblastoma so as to improve the efficacy of the hydrolysis product for treatment of the neuroblastoma. In one preferred form of the invention, the coating comprises binding molecules which are configured to target delivery of the nanoparticle to specific tissue. By way of example but not limitation, the coating for the nanoparticle comprises a monoclonal antibody against N-type calcium channels (e.g., an anti-N-type calcium channel exofacial Fab fragment) for causing the nanoparticle to bind to neural tissue (e.g., to a neuroblastoma tumor).

(35) In another form of the invention, the hydrolysis products may be delivered using a polymer system which is configured to delay degradation of the active ingredient and/or optimize the release properties of the active ingredient. By way of example but not limitation, the hydrolysis products may be “pegylated” using polyethylene glycols (PEGs) to delay premature of degradation of the active ingredient and/or optimize the release properties of the active ingredient.

(36) The selected hydrolysis products of taurolidine can be given systemically, as either a single agent or in combination with other oncolytic agents and/or radiotherapy. Examples of oncolytic agents that can be combined with the hydrolysis products of taurolidine for systemic delivery are platinum compounds (cisplatin, carboplatin), alkylating agents (cyclophosphamide, ifosfamide, melphalan, topoisomerase II inhibitor), vinca alkaloids (vincristine), and topoisomerase I inhibitors (topotecan and irinotecan).

MODIFICATIONS

(37) While the present invention has been described in terms of certain exemplary preferred embodiments, it will be readily understood and appreciated by those skilled in the art that it is not so limited, and that many additions, deletions and modifications may be made to the preferred embodiments discussed above while remaining within the scope of the present invention.