AU2019331913B2 - Taurolidine treatment for myc-expressing tumors in mammalian bodies - Google Patents

Abstract

A method for treating a cancer which overexpresses any of N-myc genes, C-myc genes and/or L-myc genes in a mammalian body, the method comprising : administering a composition to the mammalian body, wherein the composition comprises at least one from the group consisting of taurolidine; taurultam; taurinamide; methylene glycol; taurultam and taurinamide in a ratio of 1 taurultam:7 taurinamide; and taurultam, taurinamide and methylene glycol in a ratio of 1 taurultam:7 taurinamide:1 methylene glycol.

Description

WO wo 2020/047530 PCT/US2019/049266

TAUROLIDINE TREATMENT FOR MYC-EXPRESSING TUMORS IN MAMMALIAN MAMMALIAN BODIES BODIES

Applicant Applicant

CorMedix Inc.

Inventors

Bruce Reidenberg

Robert DiLuccio

Reference To Pending Prior Patent Applications

This patent application:

(i) is a continuation-in-part of pending prior

U.S. Patent U.S. Patent Application ApplicationSerial No.No. Serial 15/403,876, filedfiled 15/403,876,

01/11/2017 by CorMedix Inc. and Robert DiLuccio for

THERAPEUTIC NANOPARTICLES FOR THE TREATMENT OF

NEUROBLASTOMA AND OTHER CANCERS (Attorney's Docket No.

CORMEDIX-14), which patent application claims benefit

of prior U.S. Provisional Patent Application Serial

No. 62/277,243, filed 01/11/2016 by CorMedix Inc. and

Robert DiLuccio for NANOPARTICLE SYSTEM FOR THE

CORMEDIX-35 wo 2020/047530 WO PCT/US2019/049266

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TREATMENT OF NEUROBLASTOMA (Attorney's Docket No.

PROV) ;and CORMEDIX-14 PROV); and

(ii) claims benefit of pending prior U.S.

Provisional Patent Application Serial No. 62/725,650,

filed 08/31/2018 by CorMedix Inc. and Bruce Reidenberg

et al. for TAUROLIDINE TREATMENT FOR MYC-EXPRESSING

TUMORS IN MAMMALIAN BODIES (Attorney's Docket No.

CORMEDIX-35 CORMEDIX-35PROV). PROV) ..

The three (3) above-identified patent

applications are hereby incorporated herein by

reference.

Field Of The Invention

This invention relates to therapeutic methods and

compositions in general, and more particularly to

therapeutic methods and compositions for the treatment

of MYC-expressing tumors in mammalian bodies.

Background Of Background OfThe TheInvention Invention

Taurolidine is a well known antimicrobial with a

published mechanism of action and antimicrobial

CORMEDIX-35

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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 used as a catheter-lock

solution.

Taurolidine has recently been investigated for

oncolytic activity and found to have an 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.

MYC oncogenes have been widely described in solid

tumors and in lymphoma/leukemia.

CORMEDIX-35

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Taurolidine has demonstrated efficacy in treating

neuroblastoma in a laboratory cell line. This cell

line is known to overexpress N-myc genes.

Taurolidine has demonstrated efficacy in treating

ovarian cancer in a human ovarian cell tumor line

implanted in mice. This cell line is known to

overexpress C-myc genes.

Taurolidine has demonstrated efficacy in treating

lung cancer in a laboratory cell line. This cell line

is known to overexpress L-myc genes.

A need need exists existsfor fora anew new method method andand composition composition

which are effective which are effectiveagainst against MYC-expressing MYC-expressing tumors tumors in in

mammalian bodies.

Summary Of The Invention

In accordance with the present invention,

taurolidine, and/or the hydrolysis products of

taurolidine, is/are used to treat tumors that

overexpress N-myc genes, C-myc genes and/or L-myc

genes in mammalian bodies. Examples of tumors that

may overexpress may overexpressN-myc N-mycgenes, C-myc genes, genes C-myc and/or genes L-mycL-myc and/or

CORMEDIX-35 genes include, but are not limited to, lymphoma, melanoma, multiple myeloma, neuroblastoma, colon, breast and lung cancers.

The preferred hydrolysis products of taurolidine

may comprise at least one from the group consisting

of:

taurultam;

taurinamide;

methylene glycol;

taurultam and taurinamide in a ratio of 1

taurultam: taurultam:77taurinamide; taurinamide;and and

taurultam, taurinamide and methylene glycol in a

ratio of 1 taurultam: taurultam:77 taurinamide: taurinamide:1 1 methylene glycol.

The taurolidine 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 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,

CORMEDIX-35 CORMEDIX-35

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for an effective period of time based on individual

patient response.

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 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.

The taurultam and taurinamide (in a ratio of 1

taurultam: taurultam:77 taurinamide) taurinamide) is is given given with with aa dosage dosage range 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,

from once daily through weekly, for an effective

period of time based on individual patient response.

CORMEDIX-35

The taurultam, taurinamide and methylene glycol

(in a ratio of 1 taurultam: 7 taurinamide: 1 methylene

glycol) is glycol) is given givenwith witha a dosage range dosage of taurultam range from from of taurultam

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 of

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.

The taurolidine, and/or the hydrolysis products

of taurolidine, can be given systemically, preferably

intramuscularly or intravenously.

In one preferred form of the invention, the

taurolidine, and/or the hydrolysis products of

taurolidine, is/are delivered systemically in a

"shielded form" SO so that the taurolidine, or the

hydrolysis products of taurolidine, can reach the site

of the tumor without premature degradation, whereupon

CORMEDIX-35 the taurolidine, or the hydrolysis products of taurolidine, can treat the tumor.

More particularly,ininone More particularly, one preferred preferred formform of the of the

invention, the taurolidine and/or the hydrolysis

products of taurolidine can be delivered in the form

of a nanoparticle, where the nanoparticle comprises a

core of the taurolidine and/or the hydrolysis products

of taurolidine and an exterior coating which is

configured to prevent premature exposure of the

taurolidine and/or the hydrolysis products of

taurolidine prior to the arrival of the nanoparticle

to the tumor site. The exterior coating breaks down

as the nanoparticle travels from the site of the

insertion to the site of the tumor SO so as to release

the taurolidine and/or the hydrolysis products of

taurolidine 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.

CORMEDIX-35 CORMEDIX-35

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In another form of the invention, the taurolidine

and/or the hydrolysis products of taurolidine (i.e.,

the active ingredient) may be delivered using a

polymer system which is configured to delay premature

degradation of the active ingredient. By way of

example but not limitation, the taurolidine and/or the

hydrolysis products of taurolidine may be "pegylated"

using polyethylene glycols (PEGs) to delay premature

degradation of the active ingredient.

The taurolidine, and/or the hydrolysis products

of taurolidine, may be delivered as either a single

agent or in combination with other oncolytic agents

and/or radiotherapy.

Brief Description of Of The Drawings

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

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drawings wherein like numbers refer to like parts, and

further wherein:

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) ;

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) ;

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) ;

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

CORMEDIX-35

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from neuroblastoma but overall survival was not

significantly different from control;

Fig. 9 is a chart showing the effect of delayed

administration of a single 3-day i.p.

(intraperitoneal) bolus injection regimen of

taurolidine (20 mg/mouse/injection) on the occurrence

of i.p. human tumor xenografts in female nude mice

after after the the i.p. i.p.administration administrationof of 5 X510X 610SKOV-3 human SKOV-3 human

ovarian tumor cells;

Fig. 10 illustrates the mechanism for the

hydrolysis of taurolidine;

Fig. 11 is a chart showing the mean

pharmacokinetic parameters of taurultam; and

Fig. 12 is a chart showing the mean

pharmacokinetic parameters of taurinamide.

Detailed DescriptionOfOf Detailed Description The The Invention Invention

Taurolidine was developed as an anti-infective,

but has been found to have oncolytic activity against

neuroblastoma tumors in a laboratory cell line. This

laboratory cell line is known to overexpress N-myc

CORMEDIX-35 CORMEDIX-35 genes. More particularly, taurolidine has been found to have surprising oncolytic activity in cell cultures of human cancer cells expressing N-myc, and now in a rodent cancer model based on an N-myc expressing human cancer cell line.

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 vivo) See

Fig. 1.

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.

Furthermore, taurolidine given to CB57 SCID mice

with measurable tumors from a neuroblastoma cell line

implanted subcutaneously in the CB57 SCID mice showed

dramatic efficacy in IMR5 tumors and measurable

efficacy in SK-N-AS tumors in vivo (not in vitro) vitro)..

See Figs. 3-6.

Statistically significant decreases in tumor size

were achieved when taurolidine was administered to

CORMEDIX-35 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.

Taurolidine has also demonstrated efficacy in

treating ovarian cancer in a human ovarian cell tumor

line implanted in mice. This cell line is known to

overexpress C-myc genes. See Fig. 9 which shows the

effect of delayed administration of a single 3-day

i.p. (intraperitoneal) bolus injection regimen of

taurolidine (20 mg/mouse/injection) on the occurrence

of i.p. human tumor xenografts in female nude mice

after after the the i.p. i.p.administration administrationof of 5 X 510X 610SKOV-3 human SKOV-3 human

ovarian tumor cells. In this study, taurolidine

therapy was initiated on the day of tumor cell

inoculation or up to 5 days thereafter. Fourteen days

after the final taurolidine injection, mice in all of

the groups were sacrificed, and the peritoneal cavity

was examined for the presence of tumors. Each

experiment was experiment wasrepeated repeatedthree times, three and and times, the the pooled pooled

CORMEDIX-35 number of animals in each group ranged from 15-21

(Cancer Res. Res.,,2001 2001Sep Sep15; 15;61 61(18) (18)::6816-21, 6816-21,

Taurolidine: cytotoxic and mechanistic evaluation of a

novel antineoplastic agent, Calabresi P1, Goulette

FA, Darnowski JW).

And Taurolidinehas And Taurolidine hasalso also demonstrated demonstrated efficacy efficacy in in

treating lung cancer in a laboratory cell line. This

cell line is known to overexpress L-myc genes.

In accordance with the present invention,

taurolidine, and/or the hydrolysis products of

taurolidine, is/are used to treat tumors that

overexpress N-myc genes, C-myc genes and/or L-myc

genes in mammalian bodies. Examples of tumors that

may overexpressN-myc may overexpress N-mycgenes, genes, C-myc C-myc genes genes and/or and/or L-mycL-myc

genes include, but are not limited to, lymphoma,

melanoma, multiple myeloma, neuroblastoma, colon,

breast and lung cancers.

The mechanism for the hydrolysis of taurolidine

is shown in Fig. 10. The preferred hydrolysis

products of taurolidine that may be used to treat

tumors that overexpress N-myc genes, C-myc genes

CORMEDIX-35 CORMEDIX-35 and/or L-myc genes in mammalian bodies may comprise at least one from the group consisting of: taurultam; taurinamide; methylene glycol; taurultam and taurinamide in a ratio of 1 taurultam: taurultam:77 taurinamide; taurinamide; and and taurultam, taurinamide and methylene glycol in a ratio of 1 taurultam: taurultam:77 taurinamide: taurinamide:1 1 methylene glycol.

The taurolidine 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 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.

CORMEDIX-35

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. 12.

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.

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 with a dosage range of 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.

The taurultam, taurinamide and methylene glycol

(in a ratio of 1 taurultam: 7 taurinamide:1 taurinamide: 1methylene methylene

CORMEDIX-35 wo 2020/047530 WO PCT/US2019/049266

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glycol) glycol) is is given givenwith a dosage with range a dosage of taurultam range from from of taurultam

5 mg/kg to 280 mg/kg, with optimal range between 5

mg/kg and 40 mg/kg, from once daily through weekly,

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 of 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.

Dose selection for the hydrolysis products of

taurolidine were calculated as follows:

AUC 0-inf - Taurultam/AUC Taurultam/AUC 0-inf 0-inf Taurinamide Taurinamide = : 42.9/312.7 42.9/312.7

= 0.14. :

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

CORMEDIX-35 taurultam and taurinamide and methylene glycol in combination is 1:7:1.

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 X 1 adult

human/12 mice X 25 child BSA ratio/37 adult BSA ratio

= child dose in mg/kg

(https://www.fda.gov/downloads/drugs/guidances/ucm0789 (https://www.fda.gov/downloads/drugs/guidances/ucm0789

32.pdf)].

The taurolidine, and/or the hydrolysis products

of taurolidine, can be given systemically, preferably

intramuscularly or intravenously.

In one preferred form of the invention, the

taurolidine, and/or the hydrolysis products of

taurolidine, is/are delivered systemically in a

"shielded form" SO so that the taurolidine, or the

hydrolysis products of taurolidine, can reach the site

CORMEDIX-35 wo 2020/047530 WO PCT/US2019/049266

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of the tumor without premature degradation, whereupon

the taurolidine, or the hydrolysis products of

taurolidine, can treat the tumor.

More particularly, in one preferred form of the

invention, the taurolidine, and/or the hydrolysis

products of taurolidine, is/are delivered in the form

of a nanoparticle, where the nanoparticle comprises a

core comprising taurolidine and/or the hydrolysis

products of products oftaurolidine, taurolidine,andand an an exterior coating exterior whichwhich coating

is configured to prevent premature exposure of the

taurolidine, and/or the hydrolysis products of

taurolidine, 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 so as to release the

taurolidine, and/or the hydrolysis products of

taurolidine, intact at the site of the tumor. In one 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

CORMEDIX-35 limitation, the coating can be created from combinations of copolymers and multimers derived from polymers structured from l-lactide, 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.

If desired, the nanoparticle may comprise an

excipient (e.g.,a abuffer excipient (e.g., buffer forfor providing providing enhanced enhanced

hydrolytic stability of the taurolidine and/or

hydrolysis product within the nanoparticle). .

Additionally, if desired, the nanoparticle can

further comprise a coating, wherein the coating is

configured to target the nanoparticle to the site of a

tumor SO so as to improve the efficacy of the taurolidine

and/or hydrolysis product for treatment of the tumor.

In one preferred form of the invention, the coating

comprises binding molecules which are configured to

target delivery of the nanoparticle to specific

tissue.

CORMEDIX-35

In another form of the invention, the

taurolidine, and/or the hydrolysis products of

taurolidine, may be delivered using a polymer system

which is configured to delay premature degradation of

the taurolidine, and/or the hydrolysis products of

taurolidine, and/or to optimize the release properties

of the taurolidine, and/or the hydrolysis products of

taurolidine. By way of example but not limitation,

the taurolidine, and/or the hydrolysis products of

taurolidine, may be "pegylated" using polyethylene

glycols (PEGs) to delay premature degradation of the

taurolidine, and/or the hydrolysis products of

taurolidine, and/or to optimize the release properties

of the taurolidine, and/or the hydrolysis products of

taurolidine.

The taurolidine (and/or the hydrolysis products

of taurolidine) may be delivered as either a single

agent or in combination with other oncolytic agents

and/or radiotherapy. Examples of oncolytic agents

that can be combined with taurolidine and/or the

hydrolysis products of taurolidine for systemic

CORMEDIX-35

30 Apr 2021 2019331913 30 Apr 2021

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delivery are delivery areplatinum platinum compounds compounds (cisplatin, (cisplatin,

carboplatin), alkylating carboplatin), alkylating agents agents (cyclophosphamide, (cyclophosphamide,

ifosfamide, melphalan,topoisomerase ifosfamide, melphalan, topoisomerase II inhibitor), II inhibitor),

vinca alkaloids vinca alkaloids(vincristine), (vincristine),andand topoisomerase topoisomerase I I 2019331913

inhibitors (topotecanand inhibitors (topotecan and irinotecan). irinotecan).

Modifications Modifications

While the While the present present invention invention has has been been described described in in

terms of certain terms of certainexemplary exemplary preferred preferred embodiments, embodiments, it it

will be will be readily readilyunderstood understoodandand appreciated appreciated by those by those

skilled in the skilled in theart artthat that it it is is notnot so limited, so limited, and that and that

many additions, many additions,deletions deletions andand modifications modifications may be may be

made to made to the thepreferred preferred embodiments embodiments discussed discussed above above

while remaining while remainingwithin within the the scope scope of the of the present present

invention. invention.

Throughout thisspecification Throughout this specificationandand the the claims claims

which follow, which follow,unless unlessthe the context context requires requires otherwise, otherwise,

the word "comprise", the word "comprise",and and variations variations suchsuch as as

"comprises" and"comprising", "comprises" and "comprising", will will be be understood understood to to

imply the inclusion imply the inclusionofofa a stated stated integer integer or step or step or or

group of integers group of integersororsteps steps butbut notnot the the exclusion exclusion of of

30 Apr 2021 2019331913 30 Apr 2021

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any other integer any other integerororstep step or or group group of integers of integers or or

steps. steps.

The referencetotoany The reference anyprior prior artart in in this this

specification specification isisnot, not,and and should should notnot be taken be taken as, an as, an 2019331913

acknowledgement acknowledgement ororany any form form of of suggestion suggestion thatthat the the

prior art prior art forms formspart partofof thethe common common general general knowledge knowledge

in Australia. in Australia.

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What Is Is Claimed ClaimedIs: Is: 27 May 2025 2019331913 27 May 2025

What

1. 1. A A method method for for treating treating a a cancer cancer which which overexpresses overexpresses anyany of N-myc genes, of N-myc genes,C-myc C-mycgenes genes and/or and/or L-myc L-myc genes genes in a in a mammalian mammalian body, the method body, the methodcomprising: comprising: administering administering a acomposition compositionto to thethe mammalian mammalian body, body, wherein wherein the compositionconsists the composition consistsof of taurultam, taurultam, taurinamide taurinamide and methylene and methylene 2019331913

glycol in aaweight glycol in weightratio ratio of of 1 taurultam:7 1 taurultam:7 taurinamide:1 taurinamide:1 methylene glycol. methylene glycol.

2. 2. A A method method according according toto claim claim 1 1 further further comprising comprising administering thecomposition administering the compositionin in conjunction conjunction withwith an oncolytic an oncolytic agent and/orradiotherapy. agent and/or radiotherapy.

3. 3. A A method method according according toto claim claim 1 1 wherein wherein thethe dosage dosage range range for taurultamisisfrom for taurultam from5 5 mg/kg mg/kg to to 280280 mg/kg, mg/kg, combined combined with with taurinamide witha adosage taurinamide with dosage range range of of fromfrom 5 mg/kg 5 mg/kg to mg/kg, to 280 280 mg/kg, combined withmethylene combined with methylene glycol glycol with with a dosage a dosage range range of from of from 2.5 2.5 mg/kg to mg/kg to 160 160mg/kg, mg/kg,from from once once daily daily through through weekly, weekly, for an for an effective periodofoftime effective period time based based on on individual individual patient patient response. response.

4. 4. A A method method according according toto claim claim 3 3 wherein wherein thethe dosage dosage range range for taurultamisisoptimally for taurultam optimally from from 5 mg/kg 5 mg/kg to mg/kg, to 40 40 mg/kg, combined combined with taurinamidewith with taurinamide witha a dosage dosage range range of from of from 35 mg/kg 35 mg/kg to 40to 40 mg/kg, combined mg/kg, combinedwith withmethylene methylene glycol glycol withwith a dosage a dosage rangerange of of from from 55 mg/kg mg/kgtoto4040mg/kg, mg/kg, from from once once daily daily through through weekly, weekly, for an for an effective periodofoftime effective period time based based on on individual individual patient patient response. response.

5. 5. A A method method according according toto any any one one ofof claims claims 1-4 1-4 wherein wherein the compositionisisdelivered the composition delivered to to a patient a patient usingusing one from one from the the group consistingofofparenteral group consisting parenteral delivery, delivery, intramuscular intramuscular delivery delivery and intravenousdelivery. and intravenous delivery.

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2019331913 27 May 2025

6. 6. A A method method according according toto claim claim 1 1 wherein wherein the the composition composition is included in is included ina ananoparticle, nanoparticle,andand further further wherein wherein the the nanoparticle isconfigured nanoparticle is configuredto to delay delay exposure exposure of composition of the the composition until the nanoparticle until the nanoparticle reaches reaches thethe site site of aoftumor. a tumor.

7. 7. A A method method according according toto claim claim 6 6 wherein wherein the the nanoparticle nanoparticle 2019331913

comprises comprises aa core coreofofthe the composition composition and and an exterior an exterior coating, coating, wherein theexterior wherein the exteriorcoating coating is is configured configured to prevent to prevent exposure exposure of the composition of the compositionprior prior to to arrival arrival of the of the nanoparticle nanoparticle at the at the site of the site of the tumor. tumor.

8. 8. A A method method according according toto claim claim 7 7 wherein wherein thethe exterior exterior coating comprisesananabsorbable coating comprises absorbable polymer polymer or lipid or lipid whichwhich breaks breaks down as the down as the nanoparticle nanoparticle travels travels from from the the sitesite of insertion of insertion to to the site of the site of the thetumor. tumor.

9. 9. A A method method according according toto any any one one ofof the the preceding preceding claims claims wherein wherein the the composition composition is is delivered delivered using using aa polymer polymer system system which is configured which is configuredtoto delay delay premature premature degradation degradation of the of the composition. composition.

10. 10. AA method method according according to to claim claim 99 wherein wherein the the composition composition is “pegylated”using is "pegylated" usingpolyethylene polyethylene glycols glycols (PEGs) (PEGs) to delay to delay premature degradationofof premature degradation thethe composition. composition.

11. Use of 11. Use of aa composition composition in in the the manufacture manufacture ofof aa medicament for medicament for the the treatment treatment of of a a cancer cancer which which overexpresses overexpresses any any of N-myc genes, of N-myc genes,C-myc C-mycgenes genes and/or and/or L-myc L-myc genes genes in a in a mammalian mammalian body, wherein body, whereinthe thecomposition composition consists consists of taurultam, of taurultam, taurinamide taurinamide and methyleneglycol and methylene glycolinin a weight a weight ratio ratio of 1oftaurultam:7 1 taurultam:7 taurinamide:1 methylene taurinamide:1 methylene glycol. glycol.

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2019331913 27 May 2025

12. 12. AA composition composition forfor treating treating aa cancer cancer which which overexpresses anyofofN-myc overexpresses any N-myc genes, genes, C-myc C-myc genes genes and/or and/or L-mycL-myc genes genes in a mammalian in a mammalianbody, body,wherein wherein thethe composition composition consists consists of of taurultam, taurinamideand taurultam, taurinamide and methylene methylene glycol glycol in ain a weight weight ratioratio of of 1 taurultam:7 1 taurultam: taurinamide:1 7 taurinamide: methyleneglycol. 1 methylene glycol.

NormalNormal lymphocytes lymphocytes

TRD + Leukemia cell lines and Normal Lymphocytes Lymphocytes Normal and lines cell Leukemia + TRD Lymphocytes Normal and lines cell Leukemia + TRD MV4-11(2) MV4-11(2)

SEM(2) SEM(2)

Molt3 Molt3 SEM SEM CEM CEM

40

20

Concentration (uM) (uM) Concentration

FIG. 11 FIG.

10

1

TRD = Taurolidine Taurolidine = TRD Taurolidine = TRD 0.1 0.1

120 120 100 100 80 60 40 20 0 %survival compared to vehicle DMSO

SUBSTITUTE SHEET (RULE 26)

SK-N-BE(2) SK-N-BE(2) Taurolidine = TRD Taurolidine = TRD TRD = Taurolidine

SK-N-MC SK-N-MC SK-N-SH SK-N-SH SK-N-AS SK-N-AS

IMR5

BJ

FIG. 2

Neuroblastoma andand Neuroblastoma TRDTRD

160

microMolar in taurolidine of Concentration microMolar in taurolidine of Concentration microMolar in taurolidine of Concentration 80 80

40

20

10 TRD IC50 TRD IC50

(µM) (uM)

5 140 140 100 30 80 80 200

2.5 Neuroepithelioma Neuroepithelioma fibroblast Normal fibroblast Normal fibroblast Normal Neuroblastoma Neuroblastoma Neuroblastoma Neuroblastoma Neuroblastoma Neuroblastoma Neuroblastoma Neuroblastoma

Type

1.25 1.25

200 150 100 50 0 Cell line Cell line SK-N-BE(2) SK-N-BE(2)

% Cell viability SK-N-AS SK-N-AS SK-N-MC SK-N-SH SK-N-SH

IMR5 IMR5

BJ

SUBSTITUTE SHEET (RULE 26)

TRD by inhibition growth tumor of Evaluation : cells IMR5 TRD by inhibition growth tumor of Evaluation : cells IMR5 TRD by inhibition growth tumor of Evaluation : cells IMR5 WO 2020/047530

Taurolidine = TRD Taurolidine = TRD TRD = Taurolidine

150 Ctrl TRD TRD250mg/kg TRD 250mg/kg TRD 500mg/kg 500mg/kg

100 3/11

Tumor size(mm²) 50 I ± I

SUBSTITUTE SHEET (RULE 26) 0 30

10 20

0 implantation aftertumor Days implantation aftertumor Days implantation aftertumor Days FIG. FIG. 33 PCT/US2019/049266

= Taurolidine Taurolidine = TRD Taurolidine = TRD 100 100 750 750 500 500 250 250 500 500 00 00

inesc 0 0 0 TRD by inhibition growth tumor of Evaluation : cells IMR5 TRD by inhibition growth tumor of Evaluation : cells IMR5 ence inesc ence IMR5 cells : Evaluation of tumor growth inhibition TRD 500 by TRD Lum Lum Cou Cou

TRD nts nts

TRD 500

THE FIG. FIG. 4 4

TRD 250 TRD 250

Control Control

before before Day Day 12 12 Day 17 Day 17 After Day After Day 66 After After After After

SUBSTITUTE SHEET (RULE 26)

TRD by inhibition growth tumor of Evaluation : cells IMR5 TRD by inhibition growth tumor of Evaluation : cells IMR5 TRD by inhibition growth tumor of Evaluation : cells IMR5 Taurolidine = TRD Taurolidine = TRD Taurolidine = TRD WO 2020/047530

Ctrl TRD

8x10 TRD 250mg/kg 250mg/kg

8x10 77 TRD TRD 500mg/kg 500mg/kg

7 6x10 6x10 7

7 5/11

4x10 4x10 7

7 2x10 2x10

Tumor size (photons/second)

SUBSTITUTE SHEET (RULE 26) T 0 6 implantation aftertumor Days implantation aftertumor Days implantation aftertumor Days FIG. 5 PCT/US2019/049266

TRD by inhibition growth tumor of Evaluation : cells IMR5 TRD by inhibition growth tumor of Evaluation : cells IMR5 TRD by inhibition growth tumor of Evaluation : cells IMR5 wo 2020/047530

100 Taurolidine = TRD Taurolidine = TRD Taurolidine = TRD Control Control TRD TRDTRD 250mg/kg 250mg/kg TRD 500mg/kg 500mg/kg

50 6/11

Percent survival

SUBSTITUTE SHEET (RULE 26) 0 0 40

20 60

implantation tumor IMR5 after Days implantation tumor IMR5 after Days implantation tumor IMR5 after Days tumor) the (not animal the of survival percent is survival percent Note: tumor) the (not animal the of survival percent is survival percent Note: tumor) the (not animal the of survival percent is survival percent Note: PCT/US2019/049266

FIG. 6

*P<0.05 *P<0.05 **P<0.001 **P<0.001

*

tumor) treatment(SK-N-AS after Days tumor) treatment(SK-N-AS after Days tumor) treatment(SK-N-AS after Days FIG. 77 FIG.

* 12 * * TRD 250mg/kg TRD500mg/kg TRD 500mg/kg TRD 250mg/kg

Control Control

0 H

1 X 10 55 X X10 10 7 33 X X10 10 5 1 X 10 Taurolidine = TRD Taurolidine = TRD TRD = Taurolidine

8 1 X 10

6 1 X 10

0

Tumor size (photons/second)

SUBSTITUTE SHEET (RULE 26)

Control Control WO 2020/047530

TRD TRD250mg/kg TRD 250mg/kg TRD500mg/kg 500mg/kg

Taurolidine = TRD Taurolidine = TRD TRD = Taurolidine

50

Percent survival 8/11

0

SUBSTITUTE SHEET (RULE 26) 60

40

20

0 implantation tumor SK-N-AS after Days implantation tumor SK-N-AS after Days implantation tumor SK-N-AS after Days tumor). the (not animal the of survival percent is survival percent Note: tumor). the (not animal the of survival percent is survival percent Note: tumor). the (not animal the of survival percent is survival percent Note: FIG. FIG.88 PCT/US2019/049266

WO 2020/047530

81.8

80 60

Mice Mice with with no no tumors of evidence tumors of evidence tumors of evidence 9/11

(%) 40 18.2

20

SUBSTITUTE SHEET (RULE 26) 9.1

7.7

0 0 0

0 day

Control day 2

day 1 day 5

day 3 day 4

day0 0 injection cell tumor after initiated therapy Day injection cell tumor after initiated therapy Day injection cell tumor after initiated therapy Day PCT/US2019/049266

FIG. 9

Taurultam Taurultam

NH

HI NO2S OS Formaldehyde Formaldehyde

HCHO + H2O HCHO + H2O

+

CH2OH CHOH HNCHCHSONH + [CH(OH)] + 2H+ H2NCH2CH2SO2NH2 + CH2(OH2) ] + 2H+

Hydroxymethyl Hydroxymethyl

taurultam taurultam Methylene Methylene glycol glycol FIG. 10 FIG. 10

N OH H IN Methylene Methylene glycol glycol

O2S OS + [CH(OH)] + CH2(OH)2

-H2O -H2O +HO +H2O -H2O -HO

SO2 SO +H2O +HO

Taurinamide Taurinamide

Taurolidine Taurolidine

HINN NH Taurultam Taurultam 2HO 2H+ 2H2O

N CH2 CH Taurolidine: of Hydrolysis Taurolidine: of Hydrolysis Hydrolysis of Taurolidine:

H I NO2S N OS

HIN OS S O2

SUBSTITUTE SHEET (RULE 26)

TTmax max 0.50* 0.50* TTmax max 2.00 1.00* 1.00* 1.50 0.50 2.00 1.00 1.00 0.63 1.50 0.75 0.50

hr hr

6.9 +± 1.5 6.9 1.5 6.5 +± 1.4 6.5 1.4 6.7 +± 1.2 6.7 1.2 6.7 +± 1.3 6.7 1.3 1.1 +± 0.3 1.1 0.3 1.2 +± 0.7 1.2 0.7 1.5 +± 0.9 1.5 0.9 2.1 2.1 +± 1.1 1.1 half life half life half life half life

hr hr

53.9 53.9 +± 10.8 10.8 59.4 +± 19.4 59.4 19.4 62.6 62.6 +± 16.8 16.8 57.3 +± 15.3 57.3 15.3 51.4 +± 12.2 51.4 12.2 32.0 +± 17.9 32.0 17.9 32.0 32.0 +± 16.1 16.1 16.0 +± 4.5 16.0 4.5

Taurinamide of Parameters Pharmacokinetic Mean Taurinamide of Parameters Pharmacokinetic Mean Taurinamide of Parameters Pharmacokinetic Mean ug/mL Cmax ug/mL ug/mL Cmax

C Taurultam of Parameters Pharmacokinetic Mean Taurultam of Parameters Pharmacokinetic Mean Taurultam of Parameters Pharmacokinetic Mean C

S.D. + mean as presented are data All S.D. ± mean as presented are data All S.D. ± mean as presented are data All S.D. ± mean as presented are data All S.D. ± mean as presented are data All S.D. + mean as presented are data All FIG. 12 FIG. 11 FIG. 11 269.0 +± 35.3 269.0 35.3 310.3 +± 67.2 310.3 67.2 312.7+ ±63.2 312.7 63.2 51.8 +± 10.6 51.8 10.6 42.9 +± 11.4 42.9 11.4 356.5+ ±61.1 356.5 61.1

35.7 +± 9.0 35.7 9.0 40.7 40.7 +± 11.2 11.2

AUC(0-w) AUC(0-w) AUC(0-w) AUC(0-w) hr*ug/mL hr*ug/mL hr*ug/mL hr*ug/mL

315.4+ ±46.5 315.4 46.5 233.9 +± 29.4 233.9 29.4 271.7+±61.4 271.7 61.4 273.4 +± 55.8 273.4 55.8

37.3 37.3 +± 10.1 10.1 34.9 34.9 +± 9.6 9.6 44.8 +± 9.8 44.8 9.8 38.8 +± 9.8 38.8 9.8

AUC(0-24) AUC(0-24) hr*ug/mL hr*ug/mL hr*ug/mL hr*ug/mL

n 65 6 n 65 6 Duration of Duration of Duration of Duration of All Groups All Groups All Groups All Groups

infusion infusion infusion infusion

* = median ** == median median

0.5 0.5 0.5 hr hr 21 21

SUBSTITUTE SHEET (RULE 26)

Claims (12)

What Is Claimed Is:

1. A method for treating a cancer which overexpresses any

of N-myc genes, C-myc genes and/or L-myc genes in a mammalian

body, the method comprising:

administering a composition to the mammalian body, wherein

the composition consists of taurultam, taurinamide and methylene

glycol in a weight ratio of 1 taurultam:7 taurinamide:1

methylene glycol.

2. A method according to claim 1 further comprising

administering the composition in conjunction with an oncolytic

agent and/or radiotherapy.

3. A method according to claim 1 wherein the dosage range

for taurultam is from 5 mg/kg to 280 mg/kg, combined with

taurinamide with a dosage range of from 5 mg/kg to 280 mg/kg,

combined with methylene glycol with a dosage range of from 2.5

mg/kg to 160 mg/kg, from once daily through weekly, for an

effective period of time based on individual patient response.

4. A method according to claim 3 wherein the dosage range

for taurultam is optimally from 5 mg/kg to 40 mg/kg, combined

with taurinamide with a dosage range of from 35 mg/kg to 40

mg/kg, combined with methylene glycol with a dosage range of

from 5 mg/kg to 40 mg/kg, from once daily through weekly, for an

effective period of time based on individual patient response.

5. A method according to any one of claims 1-4 wherein

the composition is delivered to a patient using one from the

group consisting of parenteral delivery, intramuscular delivery

and intravenous delivery.

- ZD -

6. A method according to claim 1 wherein the composition

is included in a nanoparticle, and further wherein the

nanoparticle is configured to delay exposure of the composition

until the nanoparticle reaches the site of a tumor.

7. A method according to claim 6 wherein the nanoparticle

comprises a core of the composition and an exterior coating,

wherein the exterior coating is configured to prevent exposure

of the composition prior to arrival of the nanoparticle at the

site of the tumor.

8. A method according to claim 7 wherein the exterior

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.

9. A method according to any one of the preceding claims

wherein the composition is delivered using a polymer system

which is configured to delay premature degradation of the

composition.

10. A method according to claim 9 wherein the composition

is "pegylated" using polyethylene glycols (PEGs) to delay

premature degradation of the composition.

11. Use of a composition in the manufacture of a

medicament for the treatment of a cancer which overexpresses any

of N-myc genes, C-myc genes and/or L-myc genes in a mammalian

body, wherein the composition consists of taurultam, taurinamide

and methylene glycol in a weight ratio of 1 taurultam:7

taurinamide:1 methylene glycol.

12. A composition for treating a cancer which

overexpresses any of N-myc genes, C-myc genes and/or L-myc genes

in a mammalian body, wherein the composition consists of

taurultam, taurinamide and methylene glycol in a weight ratio of

1 taurultam:7 taurinamide:1 methylene glycol.