WO2019051548A1

WO2019051548A1 - Treatment of neuro-inflammation

Info

Publication number: WO2019051548A1
Application number: PCT/AU2018/050997
Authority: WO
Inventors: Graham Kelly; Benny EVISON
Original assignee: Norbio No. 2 Pty Ltd
Priority date: 2017-09-15
Filing date: 2018-09-13
Publication date: 2019-03-21

Abstract

The invention relates to treating or minimising neuro-inflammation in an individual requiring said treatment utilising a compound according to general formula (I), (II), (III), (IV) and/or (V) described herein.

Description

Treatment of neuro-inflammation

Field of the invention

The invention relates to treatment of diseases or conditions associated with neuro-inflammation including acute injury and chronic neural diseases and conditions of the CNS.

Background of the invention

Reference to any prior art in the specification is not an acknowledgment or suggestion that this prior art forms part of the common general knowledge in any jurisdiction or that this prior art could reasonably be expected to be understood, regarded as relevant, and/or combined with other pieces of prior art by a skilled person in the art.

Neuro-inflammation is fundamentally a protective cellular response aimed at removing injurious stimuli and initiating the healing process. These stimuli may arise from acute or chronic injury, and it is in this context that neuro-inflammation is generally understood as a key element in the pathophysiology of, and specific to, a wide range of conditions of the central nervous system (CNS).

Neuro-inflammation may accompany acute injury in the form of invasive injury of CNS tissue, neurotoxins, bacterial or viral infiltration of the CNS tissue, or ischaemia- reperfusion injury. Stroke and traumatic brain injury are characterized by a neuro- inflammatory response in which microglia activation and macrophage/neutrophil infiltration are important elements. If left unchecked, stroke associated neuro- inflammation can ultimately lead to secondary injury [Skaper S D et al. 2103 Immunology 141 :314-3271.

Chronic conditions that are associated with neuro-inflammation of the CNS include Alzheimer's disease, Parkinson's disease, multiple sclerosis, epilepsy and motor neuron disease. The beta-amyloid plaques and neurofibrillary tangles that are characteristic of Alzheimer's disease and related plaque -containing neurodegenerative conditions are the likely cause of the associated neuro-inflammation. Treatment with anti-neuro-inflammatory drugs has been shown to minimize risk, delay onset or slow progression of disease [Tassoni D et al. 2008 Asia Pac AJ Clin Nutri 17(S1 ):220-2281.

The prolonged exposure of CNS tissue to pro-inflammatory mediators produced principally by glial cells is understood to result in an overriding of the boundary of a normal physiological control leading to a destructive neuro-inflammatory response. Glia and microglial are confined to the CNS and it is understood that in this anatomical compartment they interact with a variety of cells of non-neural origin, principally in response to pro-inflammatory signals received from immune cells such as mast cells. [Skaper et al. supra] This cellular network which is unique to the CNS suggests pro and anti- inflammatory mechanisms of action that are specific to the CNS [Cherry J D et al. 2014 J. Neuroinflammation 1 1 :981.

Eicosanoids are but one family of molecules among the plethora of proinflammatory mediators that are considered to be involved in neuro-inflammation in the CNS. Examples include prostaglandins, prostacyclins, leukotrienes and thromboxane A₂ (TXA₂). The place of these eicosanoid molecules in the inflammatory cascade is summarised in Figure 1 . The prostaglandins include metabolites of arachidonic acid such as PGG2, PGH2, PGE2, PGD2, PGF2, PGA2 and PGB2. The prostacyclins include metabolites such as PG12. The leukotrienes include metabolites such as LTA4, LTB4, LTC4, LTD4 and LTE4. The thromboxanes include the inactive metabolite thromboxane B₂ (TXB₂). Within each of the first 3 categories (prostaglandins, prostacyclins and leukotrienes) there are different sub-families of metabolites, many of which have diametrically opposed biological functions. This serves to highlight the complex equilibrium that underlies the inflammatory process, which itself serves to highlight the inherent drawbacks of developing drugs to treat chronic inflammation without taking into consideration the effect of inhibiting some pathways on this complex interplay.

Some eicosanoids have been observed to have pro-inflammatory effect in some neural tissue and anti-inflammatory effect in non-neural tissue. Prostaglandin E 2 (PGE₂) is one example iCudaback E. et. al. 2014 Biochem Pharmacol 88: 565-572: Howes L G et al. 2007 Expert Opin. Investiq. Drugs 16(8):1 255-12661. It is believed that these effects are in part a consequence of the multiple receptor types that can be bound by a specific eicosanoid ligand, and the cells on which those receptors are expressed. Eicosanoids are thought to arise in injured tissue as a consequence of phospholipase A₂ and/or phospholipase C on activity mediated by glial cell cytokine and chemokine release. Glial cells are activated in response to CNS specific injury mentioned above. PLA₂ and PLC activity results in diacylglycerol or phospholipid hydrolysis leading to arachidonic acid production [Tassoni supra].

Arachidonic acid is a substrate enabling the formation of metabolites for a large variety of enzymes that catalyse the formation of prostaglandins, thromboxane and leukotrienes. The amount of each type of eicosanoid is generally a function of the specific activity of each enzyme in the relevant local CNS region. The targeting of a specific enzyme can lead to inhibition of formation of a metabolite and/or shunting of metabolites to another enzymatic pathway leading to the production or inhibition of production of another eicosanoid metabolite.

Some enzymes that result in inflammatory eicosanoid have been targeted for the purpose of minimizing neuro-inflammation and the focus has largely been on inhibiting COX-2 activity so as to minimize PGH₂ production. Some inhibitors that have been trialed in the treatment of Alzheimer's disease include COX-2 selective NSAIDs. It is believed that COX-2 inhibition may not be ideal as this could ablate the production of a range of prostaglandins and thromboxanes, possibly leading to shunting of arachidonic acid into the lipoxygenase pathway [Cudaback supra]. It is not known if inhibition of any enzymes downstream of COX-2 could result in at least a minimization of neuro- inflammation.

There are 3 main classes of anti-inflammatory drugs - corticosteroids, nonsteroidal drugs, and specific inhibitors.

Cortisone and prednisone are the body's natural anti-inflammatory hormones. Synthetic analogs include dexamethasone, hydrocortisone and prednisolone. Steroidal anti-inflammatories are the most effective therapies for most cases of chronic inflammation. They work predominantly by suppressing genes that have been upregulated by proinflammatory transcription factors. These genes encode for the main proinflammatory factors - cytokines, chemokines, adhesion molecules, inflammatory enzymes, receptors and proteins. While it generally is accepted that the corticosteroids provide the most potent and the best form of controlling inflammation in a way that is least likely to disrupt the equilibrium between multiple interrelated aspects of the inflammatory cascade, their use is limited due to a range of side-effects associated with chronic dosing including suppression of the immune system, water retention (oedema), sleeplessness, and mental disorders. These side-effects mean that corticosteroid use generally is limited to short periods of treatment.

The non-steroidal anti-inflammatory drugs (NSAIDs) are the most commonly prescribed anti-inflammatory drugs. Although not as potent as the corticosteroids and less forgiving of the equilibrium between different aspects of the inflammatory cascade, they nevertheless have fewer side-effects compared to the corticosteroids and therefore are suitable for chronic administration. Aspirin, indomethacin, ibuprofen, paracetamol and diclofenac are examples of the most commonly-used NSAIDs. They work by blocking enzymes known as cyclooxygenase 1 and cyclooxygenase 2 (COX 1 and 2 inhibitors), blocking the production of the majority of arachidonic metabolites (prostaglandins, prostacyclins, leukotrienes and thromboxanes). While NSAIDs are without the serious side-effects associated with corticosteroid use, their primary drawback is that they are generally regarded as a blunt instrument, blocking all eicosanoid synthesis regardless of the individual nature of the inflammatory process. This disrupts the equilibrium between opposing eicosanoids, depriving the body of beneficial functions, such as the role of prostacyclins in protecting the lining of the gut from acidic digestive juices.

The third class of anti-inflammatory drugs are those designed to target single proinflammatory factors, whether they be eicosanoids or any one of the myriad of non- eicosanoid chemical messengers such as the interleukins. The use of such drugs typically is limited to specific inflammatory conditions where a single chemical factor is known to be prominent. An example is the drug adalimumab (Humira™) which is a specific inhibitor of TNFa and is used in the treatment of inflammation associated with rheumatoid arthritis. Another example includes drugs designed to block the production of thromboxane A₂ or to block its ability to bind to its receptor. WO200610821 2 discloses the use of isoflavonoid compounds for the treatment of systemic inflammatory diseases, particularly inflammatory bowel disease, and demonstrates that the compounds selectively inhibit thromboxane A2 synthase. There are three main challenges to the successful treatment of chronic inflammation of the CNS: (i) the uniqueness of inflammation of CNS tissue compared to systemic inflammation; (ii) the likely idiosyncratic nature of the inflammatory process accompanying each specific disease state; and (iii) the blood-brain barrier (BBB). In terms of the uniqueness of inflammation of CNS tissue, as described above,

CNS tissue is comprised of unique cellular networks, which modulate pro- and antiinflammatory mediators specific to the CNS. Further, as described above, some eicosanoids have been observed to have pro-inflammatory effect in some neural tissue and anti-inflammatory effect in non-neural tissue. In terms of the ability of the drug to gain access to CNS tissue, another challenge with treating inflammation of CNS tissue as compared to non-neural inflammation is that for drug targets in the CNS, the BBB can be problematic because of the potential to restrict or prevent drugs from reaching their site of action. The BBB is a structure composed of endothelial cells that form a barrier between the brain and circulating blood. Endothelial cells form tight junctions that strictly regulate what may pass the BBB and enter the interstitial space. Cortisone has been found to cross the BBB but at very low levels, which are insufficient to exert a therapeutic effect.

There is therefore a need for improved treatments of diseases or conditions associated with inflammation of CNS tissue by addressing one or more of the following matters:

• The ability of the drug to access the CNS at levels that are therapeutically relevant.

• The ability to target the key components of the inflammatory process in the specific CNS disorder and to do so with reduced disruption to the homeostasis of the CNS.

• The ability to successfully treat the inflammatory process in CNS tissue with reduced disruption to the homeostasis of the rest of the body.

Summary of the invention

In one embodiment there is provided a method for treating or minimising neuro- inflammation of the CNS in an individual requiring said treatment including: (i) providing an individual for whom treatment or minimisation of neuro- inflammation is required;

(ii) administering a therapeutically effective amount of a compound of general formula (I) to the individual

(I) wherein D is halo;

R-i is H, C-i-10 alkyl, aryl, arylalkyl or R_ACO where R_A is Ci_-10 alkyl or an amino acid;

R₂ is H, OH or OR₆;

R₃ is H, OH, or R_B where RB is an amino acid or COR_A where R_A is as previously defined;

A and B together with the atoms between them form a six membered ring selected from the group

wherein R₄ is H, CORQ where R_D is H , OH , Cn₀ alkyl or an amino acid, CO₂Rc where R_c is C-i -10 alkyl, CORE where RE is H, Cno alkyl or an amino acid, COOH, CORc where Rc is as previously defined, or CON H RE where RE is as previously defined;

R₅ is H, CO₂Rc where R_c is as previously defined, or COR_COR_E where R_c and RE are as previously defined, and where the two R₅ groups are attached to the same group they are the same or different, or

R₅ is substituted or unsubstituted aryl or substituted or unsubstituted heteroaryl;

R₆ is Cno alkyl, aryl or arylalkyl;

X is O, N or S;

Y is

where R₇A and R₇B are each independently H, OH or OR₇; with the proviso that at least one of R₇A and R₇B is OH or OR₇; wherein R₇ is C1 -10 alkyl, aryl or arylalkyl; and

" == " represents either a single bond or a double bond; wherein said administration minimises or treats neuro-inflammation in the individual.

In one embodiment there is provided a method for treating or minimising neuro- inflammation in an individual requiring said treatment including:

(i) providing an individual for whom treatment or minimisation of neuro- inflammation is required;

(ii) administering a therapeutically effective amount of a compound of general formula (II) to the individual:

(II) wherein D is halo;

Ri is H, or RACO where RA is CM O alkyl or an amino acid; F?2 is H, OH, or RB where RB is an amino acid or CORA where RA is as previously defined;

A and B together with the atoms between them form the group:

wherein

R₄ is H, CORD where RD is H , OH , C_no alkyl or an amino acid, C0₂Rc where Rc is Cno alkyl, COR_E where R_E is H, Cno alkyl or an amino acid, COOH, COR_C where Rc is as previously defined, or CON H R_E where R_E is as previously defined;

X is O, N or S;

Y is

where R₇ is H , or C- O alkyl; and

" == " represents either a single bond or a double bond; wherein said administration minimises or treats neuro-inflammation individual.

In one embodiment there is provided a method for treating or minimising neuro- inflammation in an individual requiring said treatment including: (i) providing an individual for whom treatment or minimisation of neuro- inflammation is required;

(ii) administering a therapeutically effective amount of a compound of general formula (I II) to the individual:

wherein

D is halo; represents either a single bond or a double bond; R is H, C _{i 0} alkyl, aryl, arylalkyl or R_ACO where R_A is C _-10 alkyl or an amino acid;

R₅ is O when == is a double bond, or R₅ is hydrogen or hydroxyl when ===== is a single bond;

Y is

where R_7A and R_7B are each independently H, OH or OR₇; with the proviso that at least one of R_7A and R_7B is OH or OR₇; and wherein R₇ is Ci-i₀ alkyl, aryl or arylalkyl; wherein said administration minimises or treats neuro-inflammation in the individual.

(ii) administering a therapeutically effective amount of a compound of general formula (IV) to the individual:

(IV) wherein

D is halo, and wherein said administration minimises or treats neuro-inflammation in the individual.

(ii) administering a therapeutically effective amount of a compound of general formula (V) to the individual:

(V) wherein

(1 )

In some aspects of the present invention, the compounds may be present as racemic mixtures. In other aspects, specific enantiomers will be favoured. In one embodiment, therefore, the present invention utilises one of the following enantiomers:

In one embodiment, the compound of the present invention is

In another embodiment, the compound of the present invention is

In another embodiment there is provided a compound of any one of general formula (I), (II), (III), (IV) and/or (V), preferably compound (1 ), for use in a method of minimising or treating neuro-inflammation in an individual. In another embodiment, there is provided a compound of any one of general formula (I), (II), (III), (IV) and/or (V), preferably compound (1 ), for use in the manufacture of a medicament for minimising or treating neuro-inflammation in an individual.

In the above described embodiments, the neuro-inflammation may be associated with ischaemia or reperfusion injury such as stroke or traumatic brain injury. In the above described embodiments, the neuro-inflammation may be associated with a neurodegenerative disorder, such as Alzheimer's disease, Parkinson's disease, motor neurone disease or multiple sclerosis.

In the above described embodiments, a compound of any one of general formula (I), (II), (III), (IV) and/or (V), preferably compound (1 ), may inhibit one or more pro- inflammatory mediators including, but not limited to, kinases and cytokines. Preferably, the pro-inflammatory mediator is selected from the group consisting of: IRAK4, PCSK9, leptin, IL-1 a, TNFa, MMP-9, CCL22, CCL2, Syk, thromboxane A₂, and a combination thereof. In a particularly preferred embodiment, the pro-inflammatory mediator is IRAK4.

In one embodiment, there is provided a pharmaceutical composition including a compound of any one of general formula (I), (II), (III), (IV) and/or (V), preferably compound (1 ), for use in an IRAK4 associated condition or disease of the CNS.

In another embodiment, there is provided a method for treating an individual having an IRAK4 associated condition or disease of the CNS including administering to the individual a pharmaceutical composition including a compound of any one of general formula (I), (II), (III), (IV) and/or (V), preferably compound (1 ). In another embodiment, there is provided use of a compound of any one of general formula (I), (II), (III), (IV) and/or (V), preferably compound (1 ), in the manufacture of a medicament for treating an individual having an IRAK4 associated condition or disease of the CNS. Preferably, the composition is provided in an amount effective to minimise

IRAK4-mediated inflammatory activity.

Preferably, the IRAK4 associated condition or disease of the CNS is selected from the group consisting of: Alzheimer's disease, Parkinson's disease, motor neurone disease or multiple sclerosis. In the above described embodiments, the IRAK4 associated condition or disease of the CNS may be associated with ischaemia or reperfusion injury such as stroke or traumatic brain injury.

In one embodiment, in the above described embodiments, the compound may be administered rectally. In another embodiment there is provided a composition including:

- an oleaginous base for use in a device for rectal, vaginal or urethral application;

- a compound of general formula (I), (II), (III), (IV) or (V).

Brief description of the figures

Figure 1 : Schematic of the eicosanoid inflammatory cascade. Figure 2: Mean plasma, brain, and rectum concentration-time profile of compound (1 ) after rectal dosing of compound (1 ) at 100mg/kg (API) in male SD rats (N=3/arm).

Figure 3: Mean plasma concentration-time profile of compound (1 ) and its glucuronide metabolite following rectal administration of compound (1 ) at 1 00mg/kg (API) in male SD rats (N=3/arm). Detailed description of the embodiments

Reference will now be made in detail to certain embodiments of the invention. While the invention will be described in conjunction with the embodiments, it will be understood that the intention is not to limit the invention to those embodiments. On the contrary, the invention is intended to cover all alternatives, modifications, and equivalents, which may be included within the scope of the present invention as defined by the claims.

One skilled in the art will recognize many methods and materials similar or equivalent to those described herein, which could be used in the practice of the present invention. The present invention is in no way limited to the methods and materials described.

It will be understood that the invention disclosed and defined in this specification extends to all alternative combinations of two or more of the individual features mentioned or evident from the text. All of these different combinations constitute various alternative aspects of the invention.

As used herein, except where the context requires otherwise, the term "comprise" and variations of the term, such as "comprising", "comprises" and "comprised", are not intended to exclude further additives, components, integers or steps. A. Compounds & synthetic methods

As discussed above, in one aspect, the invention provides for uses of isoflavonoid compounds for minimising or treating neuro-inflammation. These compounds and relevant synthetic methods are described below.

In one embodiment, the compound for use in the method of minimising or treating neuro-inflammation is a compound of general formula (I)

(I) wherein D is halo;

Ri is H , C-i -10 alkyl, aryl, arylalkyi or R_ACO where R_A is C1 -10 alkyl or an amino acid;

R₂ is H, OH or OR₆;

R₃ is H, OH, or R_B where RB is an amino acid or CORA where RA is as previously defined;

or A is -OR6 and B is:

wherein

R₄ is H, CORD where RD is H, OH, C_no alkyl or an amino acid, CO₂Rc where Rc is Cno alkyl, CORE where RE is H, Cno alkyl or an amino acid, COOH, CORc where Rc is as previously defined, or CONHR_E where RE is as previously defined;

R₅ is H, CO₂Rc where Rc is as previously defined, or CORCORE where Rc and R_E are as previously defined, and where the two R₅ groups are attached to the same group they are the same or different, or

R5 is substituted or unsubstituted aryl or substituted or unsubstituted heteroaryl;

R₆ is Cno alkyl, aryl or arylalkyl;

X is O, N or S;

Y is

where R₇A and R₇B are each independently H, OH or OR₇; with the proviso that at least one of R₇A and R₇B is OH or OR₇; wherein R₇ is C₁-₁0 alkyl, aryl or arylalkyl; and

" == " represents either a single bond or a double bond;

In one particularly preferred embodiment of general formula (I), D is bromo. In one embodiment, A and B together with the atoms between them form the six membered ring:

In one embodiment, Ri is H.

In one embodiment, R₂ IS H.

In one embodiment, R₃ is H.

In one embodiment, R is H.

In one preferred embodiment, R₇A is H and R₇B is OH.

In one embodiment, the compound for use in the method of minimising or treating neuro-inflammation is a compound of general formula (II)

(II) wherein

D is halo;

Ri is H, or RACO where RA is C1-10 alkyl or an amino acid; F?2 is H, OH, or RB where RB is an amino acid or CORA where RA is as previously defined;

A and B together with the atoms between them form the group:

wherein

R₄ is H, CORD where RD is H , OH , C_no alkyl or an amino acid, C0₂Rc where Rc is R_C is Cno alkyl COR_E where R_E is H , C_NO alkyl or an amino acid, COOH, COR_C where Rc is as previously defined, or CONH R_E where R_E is as previously defined;

X is O, N or S;

Y is

where R₇ is H, or CM O alkyl; and

" == " represents either a single bond or a double bond. In one particularly preferred embodiment, D is bromo. In one embodiment, R₂ is H . In one embodiment, R₄ is H . In one embodiment, R₇ is H . In another embodiment, the compound for use in the method of minimising or treating neuro-inflammation is a compound of general formula (I II)

- a compound of general formula (I II) :

wherein

D is halo;

" represents either a single bond or a double bond;

Ri is H , C-i -10 alkyl, aryl, arylalkyl or RACO where RA is C1 -10 alkyl or an amino acid;

R₅ is O when == is a double bond, or R₅ is hydrogen or hydroxyl when single bond;

Y is

where R₇A and R₇B are each independently H, OH or OR₇; with the proviso that at least one of R_7A and R_7B is OH or OR₇; and wherein R₇ is Ci_-10 alkyl, aryl or arylalkyl.

In one particularly preferred embodiment, D is bromo. In one embodiment, D is in the 8-position as shown below:

In one embodiment, Ri is H.

In one preferred embodiment, is a double bond and R₅ is O.

In one preferred embodiment, R₇A is H and R₇B is OH.

(IV) wherein

D is halo, and wherein said administration minimises or treats neuro-inflammation ndividual.

Preferably, D is bromo. In one embodiment there is provided a method for treating or minimising neuro- inflammation in an individual requiring said treatment including:

(i) providing an individual for whom treatment or minimisation of neuro- inflammation is required; (ii) administering a therapeutically effective amount of a compound of general formula (V) to the individual:

wherein

Preferably, D is bromo.

In one particularly preferred embodiment, the compound is compound (1 )

(1 )-

In one embodiment, there is provided a method for treating an individual having an IRAK4 associated condition or disease of the CNS including administering to the individual a pharmaceutical composition including a compound of any one of general formula (I), (II), (III), (IV) and/or (V), preferably compound (1 ). Preferably, the composition is provided in an amount effective to minimise IRAK4 inflammatory activity. As used herein the term "alkyl" refers to a straight or branched chain hydrocarbon radical having from one to ten carbon atoms, or any range between, i.e. it contains 1 , 2, 3, 4, 5, 6, 7, 8, 9 or 1 0 carbon atoms. The alkyl group is optionally substituted with substituents, multiple degrees of substitution being allowed. Examples of "alkyl" as used herein include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, n-pentyl, isopentyl, and the like.

As used herein, the term "CMO alkyl" refers to an alkyl group, as defined above, containing at least 1 , and at most 1 0 carbon atoms respectively, or any range in between (e.g. alkyl groups containing 2-5 carbon atoms are also within the range of Ci - io).

Preferably the alkyl groups contain from 1 to 5 carbons and more preferably are methyl, ethyl or propyl.

As used herein, the term "aryl" refers to an optionally substituted benzene ring. The aryl group is optionally substituted with substituents, multiple degrees of substitution being allowed.

As used herein, the term "heteroaryl" refers to a monocyclic five, six or seven membered aromatic ring containing one or more nitrogen, sulphur, and/or oxygen heteroatoms, where N-oxides and sulphur oxides and dioxides are permissible heteroatom substitutions and may be optionally substituted with up to three members. Examples of "heteroaryl" groups used herein include furanyl, thiophenyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, oxazolyl, isoxazolyl, oxadiazolyl, oxo- pyridyl, thiadiazolyl, isothiazolyl, pyridyl, pyridazyl, pyrazinyl, pyrimidyl and substituted versions thereof.

A "substituent" as used herein, refers to a molecular moiety that is covalently bonded to an atom within a molecule of interest. For example, a "ring substituent" may be a moiety such as a halogen, alkyl group, or other substituent described herein that is covalently bonded to an atom, preferably a carbon or nitrogen atom, that is a ring member. The term "substituted," as used herein, means that any one or more hydrogens on the designated atom is replaced with a selection from the indicated substituents, provided that the designated atom's normal valence is not exceeded, and that the substitution results in a stable compound, i.e., a compound that can be isolated, characterised and tested for biological activity.

The terms "optionally substituted" or "may be substituted" and the like, as used throughout the specification, denotes that the group may or may not be further substituted, with one or more non-hydrogen substituent groups. Suitable chemically viable substituents for a particular functional group will be apparent to those skilled in the art.

Examples of substituents include but are not limited to:

C-|-C₆ alkyl, C-| -C₆ haloalkyl, C-| -C₆ haloalkoxy, C-|-C₆ hydroxyalkyl, C3-C7 heterocyclyl, C3-C7 cycloalkyl, C_rC₆ alkoxy, CrC₆ alkylsulfanyl, CrC₆ alkylsulfenyl, C C6 alkylsulfonyl, C1-C6 alkylsulfonylamino, arylsulfonoamino, alkylcarboxy, alkylcarboxyamide, oxo, hydroxy, mercapto, amino, acyl, carboxy, carbamoyl, aminosulfonyl, acyloxy, alkoxycarbonyl, nitro, cyano or halogen.

The term "isoflavonoid" as used herein is to be taken broadly and includes isoflavones, isoflavenes, isoflavans, isoflavanones, isoflavanols and similar or related compounds. Some non-limiting examples of isoflavonoid core structures are shown below:

wherein " " represents either a single bond or a double bond.

Methods for synthesis of isoflavonoid type compounds are described W01998/008503 and WO2005/049008 and references cited therein towards synthesis, the contents of which are incorporated herein by reference in entirety. Methods for the synthesis of the compound below and related compounds can be found in WO2006108212.

The compounds include all salts, such as acid addition salts, anionic salts and zwitterionic salts, and in particular include pharmaceutically acceptable salts as would be known to those skilled in the art. The term "pharmaceutically acceptable salt" refers to an organic or inorganic moiety that carries a charge and that can be administered in association with a pharmaceutical agent, for example, as a counter-cation or counter- anion in a salt. Pharmaceutically acceptable cations are known to those of skilled in the art, and include but are not limited to sodium, potassium, calcium, zinc and quaternary amine. Pharmaceutically acceptable anions are known to those of skill in the art, and include but are not limited to chloride, acetate, tosylate, citrate, bicarbonate and carbonate. Pharmaceutically acceptable salts include those formed from: acetic, ascorbic, aspartic, benzoic, benzenesulphonic, citric, cinnamic, ethanesulphonic, fumaric, glutamic, glutaric, gluconic, hydrochloric, hydrobromic, lactic, maleic, malic, methanesulphonic, naphthoic, hydroxynaphthoic, naphthalenesulphonic, naphthalenedisulphonic, naphthaleneacrylic, oleic, oxalic, oxaloacetic, phosphoric, pyruvic, para-toluenesulphonic, tartaric, trifluoroacetic, triphenylacetic, tricarballylic, salicylic, sulphuric, sulphamic, sulphanilic and succinic acid. The term "pharmaceutically acceptable derivative" or "prodrug" refers to a derivative of the active compound that upon administration to the recipient is capable of providing directly or indirectly, the parent compound or metabolite, or that exhibits activity itself and includes for example phosphate derivatives and sulphonate derivatives. Thus, derivatives include solvates, pharmaceutically active esters, prodrugs or the like.

The preferred compounds of the present invention also include all derivatives with physiologically cleavable leaving groups that can be cleaved in vivo to provide the compounds of the invention or their active moiety. The leaving groups may include acyl, phosphate, sulfate, sulfonate, and preferably are mono-, di- and per-acyl oxy- substituted compounds, where one or more of the pendant hydroxy groups are protected by an acyl group, preferably an acetyl group. Typically acyloxy substituted compounds of the invention are readily cleavable to the corresponding hydroxy substituted compounds.

B. Formulations

The compounds of any one of general formula (I), (II), (III), (IV) and/or (V) may be provided in the form of a pharmaceutical composition including at least one pharmaceutically acceptable excipient, especially for use in treatment or in the manufacture of a medicament, for example, for the minimising or treating neuro- inflammation.

In another embodiment, the compounds of general formula (I), (II), (III), (IV) and/or (V) may be provided in the form of a pharmaceutical composition including at least one pharmaceutically acceptable excipient, especially for use in treatment or in the manufacture of a medicament, for example, for the minimising or treating an individual having an IRAK4 associated condition or disease of the CNS.

In the context of this application substantially pure is intended to mean 90% purity or greater such as 95% purity, particularly 98% purity, especially 99% purity, for example as assessed by HPLC analysis. The invention also extends to employing at least two compounds of general formula (I), (II), (III), (IV) and/or (V) in the various aspects of the invention described herein.

Pharmaceutical formulations include those suitable for oral, parenteral (including subcutaneous, intradermal, intramuscular, intravenous and intra-articular), inhalation (including use of metered dose pressurised aerosols, nebulisers or insufflators), intranasal, rectal and topical (including dermal, buccal, sublingual and intraocular) administration. The most suitable route may depend upon for example the condition and disorder of the recipient. The formulations may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the art of pharmacy. All methods include the step of bringing the active ingredient into association with the carrier which constitutes one or more accessory ingredients. In general the formulations are prepared by uniformly and intimately bringing into association the active ingredient with liquid carrier or finely divided solid carriers or both and then, if necessary, shaping the product into the desired formulation. Formulations of the present invention suitable for oral administration may be presented as discrete units such as capsules such as gelatine or hydroxypropyl methylcellulose (HPMC) capsules, cachets or tablets each containing a predetermined amount of the active ingredient; as a powder or granules; as a solution or a suspension in an aqueous liquid or a non-aqueous liquid; or as an oil-in-water liquid emulsion or a water-in-oil liquid emulsion. The active ingredient may also be presented as a paste.

When compounds of general formula (I), (II), (III), (IV) and/or (V) are formulated as capsules preferably the compound is formulated with one or more pharmaceutically acceptable carrier such as starch, lactose, microcrystalline cellulose, silicon dioxide and/or a cyclic oligosaccharide such as cyclodextrin. Additional ingredients may include lubricants such as magnesium stearate and/or calcium stearate.

Suitable cyclodextrins include a-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, dimethyl-p-cyclodextrin, 2-hydroxyethyl^-cyclodextrin, 2-hyroxypropyl-cyclodextrin, 3- hydroxypropyl-p-cyclodextrin and tri-rmethyl-p-cyclodextrin. More preferably the cyclodextrin is hydroxypropyl- -cyclodextrin.

Tablets may be prepared by compression or moulding, optionally with one or more accessory ingredients. Compressed tablets may be prepared by compressing in a suitable machine the active ingredient in a free-flowing form such as a powder or granules, optionally mixed with a binder, lubricant such as magnesium stearate or calcium stearate, inert diluent or a surface active/dispersing agent. Moulded tablets may be made by moulding a mixture of the powdered compound moistened with an inert liquid diluent, in a suitable machine. The tablets may optionally be coated, for example, with an enteric coating and may be formulated so as to provide slow or controlled release of the active ingredient therein. Formulations for parenteral administration include aqueous and non-aqueous sterile injection solutions which may contain antioxidants, buffers, bacteriostats and solutes which render the formulation isotonic with the blood of the intended recipient and which may include suspending agents and thickening agents. Preferably a parenteral formulation will comprise a cyclic oligosaccharide such as hydroxypropyl-β- cyclodextrin. The formulations may be presented in unit-dose or multi-dose containers, for example sealed ampoules and vials, and may be stored in a freeze-dried (lyophilised) condition requiring only the addition of the sterile liquid carrier, for example saline or water-for-injection (WFI), immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules and tablets of the kind previously described.

Dry powder compositions for topical delivery to the lung by inhalation may, for example, be presented in capsules and cartridges of for example gelatine, or blisters of for example laminated aluminium foil, for use in an inhaler or insufflator. Formulations generally contain a powder mix for inhalation of the one or more compounds of the invention and a suitable powder base (carrier substance) such as lactose or starch. Use of lactose is preferred. Each capsule or cartridge may generally contain between 20pg- 10mg of the compound formula (I) optionally in combination with another therapeutically active ingredient. Alternatively, the compound or compounds of the invention may be presented without excipients. Packaging of the formulation may be for unit dose or multi-dose delivery.

Spray compositions for topical delivery to the lung by inhalation may, for example be formulated as aqueous solutions or suspensions or as aerosols suspensions or solutions delivered from pressurised packs, such as a metered dose inhaler, with the use of a suitable liquefied propellant. Suitable propellants include a fluorocarbon or a hydrogen-containing chlorofluorocarbon or mixtures thereof, particularly hydrofluoroalkanes, e.g. dichlorodifluoromethane, trichlorofluoromethane, dichlorotetra- fluoroethane, especially 1 ,1 ,1 ,2-tetrafluoroethane, 1 ,1 ,1 ,2,3,3,3-heptafluoro-n-propane or a mixture thereof. Carbon dioxide or other suitable gas may also be used as propellant. The aerosol composition may be excipient free or may optionally contain additional formulation excipients well known in the art such as surfactants e.g. oleic acid or lecithin and cosolvents e.g. ethanol. Pressurised formulations will generally be retained in a canister (e.g. an aluminium canister) closed with a valve (e.g. a metering valve) and fitted into an actuator provided with a mouthpiece.

Medicaments for administration by inhalation desirably have a controlled particle size. The optimum particle size for inhalation into the bronchial system is usually 1 - 10pm, preferably 2-5pm. Particles having a size above 20pm are generally too large when inhaled to reach the small airways. When the excipient is lactose it will typically be present as milled lactose, wherein not more than 85% of lactose particles will have a MMD of 60-90pm and not less than 15% will have a MMD of less than 15pm.

Formulations for intranasal administration include mucoadhesive nano- emulsions. Preferably, an intranasal formulation will comprise a mucoadhesive polymer such as Chitosan, and may optionally include additives such as an oil, surfactant, cosurfactant, and combinations thereof. Suitable oils include oleic acid, which enhance transmembrane delivery. Suitable surfactants and cosurfactants include Tween 80, PEG, Labrasol, Carbitol, Tanscutol HP, Cremophore EL, Tween 20, Span 20, ethyl alcohol. Intranasal formulations may be prepared as a sterile powder or suspension of the kind previously described and may contain a preservative.

Medicaments for intranasal administration desirably have a controlled particle size. The optimum particle size for intranasal delivery is usually less than about 1 pm, preferably less than about 500 nm, more preferably less than about 200nm. It should be understood that in addition to the ingredients particularly mentioned above, the formulations of this invention may include other agents conventional in the art having regard to the type of formulation in question, for example those suitable for oral administration may include flavouring agents.

The compounds and pharmaceutical formulations according to the invention may be used in combination with or include one or more other therapeutic agents, for example anti-inflammatory agents for minimisation or treatment of neuro-inflammation. Examples may include corticosteroids and NSAIDs. Suitable corticosteroids, which may be used in combination with the compounds of the invention are those oral and inhaled corticosteroids and their pro-drugs which have anti-inflammatory activity. Examples include methyl prednisolone, prednisolone, dexamethasone, fluticasone propionate, 6a,9a-difluoro-17a-[(2-furanylcarbonyl)oxy]-l 1 β-hydroxy-l 6a-methyl-3-oxo-androsta- l,4-diene-17p-carbothioic acid S-fluoromethyl ester, 6a,9a-difluoro~l ip-hydroxy-16a- methyl-3-oxo-l 7a-propionyloxy-androsta-l ,4-diene-l 7p-carbothioic acid 5"-(2-OXo- tetrahydro-furan-3S-yl) ester, beclomethasone esters (e.g. the 17-propionate ester or the 17,21 -dipropionate ester), budesonide, flunisolide, mometasone esters (e.g. the furoate ester), triamcinolone acetonide, rofleponide, ciclesonide and butixocort propionate. Preferred corticosteroids include fluticasone propionate, and 6a,9a-difluoro- 17a-[(2-furanylcarbonyl)oxy] - 1 1 β-hydroxy- 16a-methyl-3 -oxo-androsta- 1 ,4-diene- 17 -carbothioic acid S-fluoromethyl ester, more preferably 6a,9a-difluoro-17a-[(2- furanylcarbonyl)oxy]-l ip-hydroxy-16a-methyl-3 -oxo-androsta- l,4-diene-17p-carbothioic acid S-fluoromethyl ester.

Suitable NSAIDs include sodium cromoglycate, nedocromil sodium, phosphodiesterase (PDE) inhibitors (e.g. theophylline, PDE4 inhibitors or mixed PDE3/PDE4 inhibitors), leukotriene antagonists, inhibitors of leukotriene synthesis, iNOS inhibitors, tryptase and elastase inhibitors, beta-2 integrin antagonists and adenosine receptor agonists or antagonists (e.g. adenosine 2a agonists), cytokine antagonists (e.g. chemokine antagonists) or inhibitors or cytokine synthesis.

The co-administration of active ingredients may be simultaneous or sequential. Simultaneous administration may be effected by the compounds being in the same unit dose, or in individual and discrete unit doses administered at the same or similar time. Sequential administration may be in any order as required and typically will require an ongoing physiological effect of the first or initial active agent to be current when the second or later active agent is administered, especially where a cumulative or synergistic effect is desired.

In one embodiment, the formulation is an oral formulation, more preferably a capsule formulation.

Preferably the capsule formulation will comprise consist essentially of or consist of a compound of general formula (I), (II), (III), (IV) and/or (V) and silicon dioxide.

Preferably the capsule will be a HPMC capsule. In a preferred embodiment the formulation is a suppository or enema, which can be used to direct the active ingredient more closely to the disease affected area of the body.

Formulations for rectal administration may be presented as a suppository with carriers such as cocoa butter or polyethylene glycol, or as an enema wherein the carrier is an isotonic liquid such as saline. Additional components of the formulation may include one or more surfactants, buffer salts or acid or alkali to adjust the pH, isotonicity adjusting agents and/or anti-oxidants.

In one particularly preferred embodiment, the compound of general formula (I), (II), (III), (IV) and/or (V) is provided in the form of a suppository, pessary or intra urethral device in a composition that includes an oleaginous suppository base. In this embodiment, the base is formulated so as to ensure that the bulk of the compound of general formula (I), (II), (III), (IV) and/or (V) does not partition from the base.

Generally the base has a solvent power for the compound of general formula (I), (II), (III), (IV) and/or (V) enabling at least partial, preferably complete dissolution of the isoflavonoid in the base.

The base may be comprised of, or consist of an oil or fat.

In one embodiment the base includes saturated fatty acids in an amount of 50 to 65% w/w base. Stearic acid may be included in an amount of 25 to 40% w/w base. Palmitic acid in an amount of 25 to 30% w/w base. Longer chain saturated fatty acids such as myristic, arachidic and lauric acid may be included in an amount of <2% w/w base.

In one embodiment, oleaginous bases that include unsaturated fatty acids in an amount of 35 to 50% w/w base are preferred. Monounsaturated fatty acid may be included in an amount of 30 to 45% w/w base. Oleic acid may be included in an amount of 30 to 40% w/w base. Polyunsaturated fatty acids such as linoleic and alpha linolenic acid may be included in an amount of 0 to 5% w/w base.

Theobroma oil (cocoa butter) has been a traditional base in a suppository because of: (a) its non-toxic and non-irritant nature, and (b) its low melting point, meaning that it readily dissolves at body temperature when placed within a bodily cavity, However, it is increasingly being replaced for a number of reasons. One reason is its variability in composition, a consequence of its natural origins; theobroma oil also is polymorphic, meaning it has the ability to exist in more than one crystal form. Another is that the formulated product needs to be kept refrigerated because of its low melting point, rendering it unsuitable in tropical regions. This has led to a number of substitute products offering a range of advantages over theobroma oil such as greater consistency, decreased potential for rancidity, and greater ability to tailor phase transitions (melting and solidification) to specific formulation, processing, and storage requirements. Nevertheless, theobroma oil or a fatty base with similar composition and physicochemical properties has been found to be a preferred embodiment of the invention.

Typically, the oleaginous base comprises a predominance of (>45% w/w base) of saturated fatty acids. Preferably the oleaginous base is Theobroma oil (cocoa butter) or an oil fraction or derivative or synthetic version thereof having a saturated fatty acid profile substantially the same as, or identical to the fatty acid profile of Theobroma oil.

Other examples of oils that may be used to provide or obtain fatty acids useful as bases include those obtainable from natural sources such as canola oil, palm oil, soya bean oil, vegetable oil, and castor oil. Oils derived from these sources may be fractionated to obtain oil fractions containing saturated fatty acids.

The base may be formed or derived from a hard fat, butter or tallow.

The base may comprise esterified or non-esterified fatty acid chains. The fatty acid chains may be in the form of mono, di and triglycerides, preferably of saturated fatty acid chains of C9-20 chain length. In a preferred embodiment, the base may comprise triglycerides. The base may comprise additives including PEG, PEG monoesters, PEG monostearate, PEG diesters, PEG distearate, polysorbate esters, and combinations thereof.

A suppository base may be formed from synthetic oils or fats, examples including Fattibase, Wecobee, Witepsol (IOI Oleo GmbH, Germany), Suppocire (Gattefosse, France), Hydrokote, Subanal (Dott. Bonapace, Italy) and Dehydag. The proportion of the oleaginous suppository base in the final product is a function of the dosage of active pharmaceutical ingredient and the presence of other pharmaceutical or inert ingredient (if any) but may be provided by way of example in an amount of about 1 to 99% w/w formulation. The compositions for rectal, vaginal or urethral application may be prepared as follows. The compound of general formula (I), (II), (III), (IV) and/or (V) is contacted with a suppository base (as described above) in molten form in conditions enabling at least partial, preferably complete or substantially complete dissolution of the compound of general formula (I), (II), (III), (IV) and/or (V) in the base. This solution is then poured into a suitable mould, such as a PVC, polyethylene, or aluminium mould. For example, the compound of general formula (I), (II), (III), (IV) and/or (V) may be contacted with the base at a temperature of from about 35° C to about 50° C and preferably from about 40° C to about 44° C. The compound of general formula (I), (II), (III), (IV) and/or (V) can be milled or sieved prior to contact with the base. In one embodiment, the conditions provided for manufacture, and formulation or device formed from same, enable at least, or provide at least, 50%, preferably 60%, preferably 70%, preferably 80%, preferably 90%, preferably 95% of the isoflavonoid for a given dosage unit to be dissolved in the dosage unit. In these embodiments, no more than 50% of the isoflavonoid for a given dosage unit, preferably no more than 40%, preferably no more than 30%, preferably no more than 20%, preferably no more than 10%, preferably no more than 5% of isoflavonoid for a given dosage unit may be in admixture with, (i.e. undissolved in) the suppository base of the dosage unit.

Optionally the suppositories, pessaries or intra-urethral devices may be coated, prior to packing, for example with cetyl alcohol, macrogol or polyvinyl alcohol and polysorbates to increase disintegration time or lubrication or to reduce adhesion on storage.

One or more sample suppositories, pessaries, or intra-urethral devices from each batch produced are preferably tested by the dissolution method of the present invention for quality control. According to a preferred embodiment, a sample from each batch is tested to determine whether at least about 75 or 80% by weight of the base dissolves within 2 hours. Typically the suppository, pessary or like device according to the invention is substantially hydrophobic or lipophilic throughout and does not contain a hydrophilic substance such as hydrophilic carrier or pharmaceutical active, or hydrophilic foci or region formed from the ligation or complexing of the isoflavonoid to or with another pharmaceutical compound, carrier or excipient.

The total weight of the suppository preferably ranges from about 1500 mg to about 3000 mg, preferably 1750 mg to about 2500 mg. In another embodiment the total weight of the suppository preferably ranges from about 2250 mg to about 2700 mg, and more preferably from about 2250 to about 2500 mg. According to one embodiment, the suppository has a total weight ranging from about 2300 mg to about 2500 mg.

The suppository or pessary is preferably smooth torpedo-shaped.

The melting point of the suppository or pessary is generally sufficient to melt in the patient's body, and is typically no more than about 37° C.

In one particularly preferred embodiment there is provided:

- a kit including: a plurality of suppositories sufficient in number to provide an individual with a suppository once daily, or twice daily, for a period of 30 to 90 days, preferably 30 to 60 days, preferably 30 days each suppository including:

400mg, 800mg, or 1 ,200mg of compound (1 );

a suppository base in the form of cocoa butter; wherein the suppository base in provided an amount of 1 -99% w/w of the suppository, - the kit further including written instructions to provide the suppository once daily, or twice daily for a period of 30 to 90 days, preferably 30 to 60 days, preferably 30 days.

C. Indications & treatment methods The term "neuro-inflammation," unless otherwise indicated herein, refers to inflammation of the nervous tissue of the central nervous system (i.e. brain, spinal cord). Neuro-inflammation can be caused by any means, including but not limited to diseases, disorders, infection, exposure to LPS, traumatic brain injury, toxic metabolites, ischaemia/reperfusion injury or autoimmunity. In certain instances, activation of glial cells (e.g., astrocytes, microglia, or macroglia) can be part of the cause of or result from neuro-inflammation. In other instances, a compromised blood-brain-barrier (BBB) contributes to or is the result of neuro-inflammation.

Neuro-inflammation can be acute, chronic, or both. Acute neuro-inflammation can, in certain instances, be caused by any means, such as a disease, a disorder, a result of injury including ischaemia/reperfusion injury, or induced peripheral stimuli. Acute neuro-inflammation can, for example, result in platelet deposition, tissue edema, endothelial cell activation, or an increase in certain inflammatory-related molecules (e.g., chemokines or cytokines). Chronic neuro-inflammation can, in certain instances, be caused by any means, such as a disease, a disorder, a result of one or more of neurodegenerative diseases, aging, autoimmunity, viruses, microbes (e.g., bacteria or yeast), traumatic injury, traumatic brain injury, toxic metabolites, or induced peripheral stimuli. Chronic neuro-inflammation can include one or more of sustained activation of glial cells, recruitment of other immune cells into the brain.

In some forms of neuro-inflammation, there is activation (in some instances it is a rapid activation) of microglia (e.g., astrocytes), but little or no peripheral immune response. In other forms, microglia produce oxidative species and there is a peripheral immune response (e.g., recruiting peripheral immune cells). In other instances, microglia (e.g., astrocytes) can become activated, can release a variety of growth factors, can undergo morphological changes, or combinations thereof. Astrocytes can in certain instances be activated and such activation can, for example provide biochemical support of endothelial cells that form the blood-brain barrier, provide nutrients to the nervous tissue, maintain extracellular ion balance, assist in the repair and scarring process of the brain and spinal cord (e.g., following traumatic injuries or acute inflammation), support neurons, and assist in synaptic plasticity mechanisms.

In some forms of neuro-inflammation, cytokines (e.g., chemokines) can be released by brain cells (e.g., glial cells, microglia, astrocytes, or endothelial cells), up- regulated or down-regulated by brain cells, or can have an increased or decreased level in the brain. Such cytokines can include, but are not limited to pro-inflammatory cytokines (e.g., IL-6, IL-la, IL-Ιβ, TNF- (Tumor Necrosis factor-alpha), or CCL2 (Chemokine ligand 2)) or anti-inflammatory cytokines. In certain instances, the release of one or more cytokines can result in a compromised BBB. In certain aspects, the level of pro-inflammatory cytokines is increased or up-regulated. In other aspects, the level of anti-inflammatory cytokines is decreased or down-regulated.

The term "minimising neuro-inflammation," unless otherwise indicated herein, refers to diminishing one or more causes of, manifestations of, or results of neuro- inflammation (acute or chronic), such as but not limited to activation of glial cells, compromised BBB, increased levels of pro-inflammatory cytokines (e.g., chemokines), or decreased levels of anti-inflammatory cytokines.

The term "preventing neuro-inflammation," unless otherwise indicated herein, refers to stopping or diminishing the potential onset of one or more causes of manifestations, or results of neuro-inflammation (acute or chronic), such as but not limited to activation of glial cells, compromised BBB, increased levels of proinflammatory cytokines (e.g., chemokines), or decreased levels of anti-inflammatory cytokines.

The term "treating neuro-inflammation," unless otherwise indicated herein, refers to reducing neuro-inflammation, ameliorating at least one symptom resulting from neuro-inflammation (such as a symptom of a disease or disorder that could cause neuro-inflammation), or both.

In a particularly preferred embodiment, there is provided a method of minimising or treating acute neuro-inflammation arising from, or associated with reperfusion/ischaemia related injury such as stroke, and traumatic brain injury. The method includes the step of administering a compound according to any one of Formula I to V, preferably compound 1 , to an individual requiring the treatment. Generally the compound is administered immediately post injury, preferably within 48 hours, preferably within 24 or 12 hours, most preferably within 6 hours or less post injury. The compound may be given intravenously. Alternatively, the compound may be given intra ventricularly. The compound may be given in an amount of about 15 to 30 mg per kg per day or less, for example, 1 0 to 20 mg per kg per day, such as 1 to 1 0 mg per kg per day of the body weight of the patient. Typically, the compound is given for a period of no longer than about 10 days, preferably 5 to 7 days or less.

In another embodiment, there is provided a method of minimising or treating acute neuroinflammation arising from or associated with neurosurgery. According to the method, a compound of any one of Formula I to V, preferably compound 1 , is administered directly to CNS tissue during or at the completion of neurosurgery thereby reducing inflammation associated with such surgery. This method may include administration directly to the tissue wherein the surgery is neurosurgery, preferably in a unit dose of between about 400 mg to about 800 mg. The compound may be given in an amount of about 15 to 30 mg per kg per day or less, for example, 1 0 to 20 mg per kg per day, such as 1 to 10 mg per kg per day of the body weight of the patient.

In another embodiment, there is provided a method of treating chronic neuro inflammation, or treating a chronic disease associated with neuroinflammation. The disease or condition may be selected from the group consisting of Alzheimer's disease, Parkinson's disease, multiple sclerosis, epilepsy and motor neuron disease. Other conditions include those characterized by neuro-fibrillar tangles and/or plaques. In these embodiments, a compound according to any one of Formula I to V, preferably compound 1 , is given one to four times daily, preferably once or twice daily and preferably so as to provide systemic delivery. A unit dose may include from 50 to 500mg of a compound of any one of Formula I to V, preferably compound 1 , more preferably 200 to 400mg, for example, about 250mg. In another embodiment, a unit does may include 400mg, 800mg, or 1 ,200mg of a compound of any one of Formula I to V, preferably compound 1 . In a particularly preferred embodiment the compound is given in the form of a suppository in a form described herein. The compound may be given in an amount of about 15 to 30 mg per kg per day or less, for example, 10 to 20 mg per kg per day, such as 1 to 1 0 mg per kg per day of the body weight of the patient. Typically, the compound is given chronically. Ultimately, the dose prescribed and intervals at which the prescribed dose is taken will be at the discretion of the physician with responsibility for the patient.

Example 1 : Suppository Formulation

To formulate the suppository, one part compound (1 ) to four parts MBK base was used. 1000 mg of compound (1 ) powder was added to 4000 mg MBK solid base. The tube was vortexed to ensure that the drug compound (1 ) was mixed well with the solid base. The tube was incubated in a water bath set at 45 °C until the base melted. The melted base was vortexed with the drug vigorously for 30 seconds followed by warming in the 45 °C water bath again for 20 seconds. This process was repeated until no clumps or undissolved particles of the drug were visible. To achieve thorough mixing of the drug in the base a 19G needle and 2 ml. syringe were used to repeatedly mix the drug by drawing it up and expelling out rapidly. The container with the base and drug were kept in a warm water bath (at 45 °C) to prevent it from solidifying. The molten base containing the drug was filled into a plastic Pasteur pipette and allowed to solidify at room temperature. This solid mass was then extruded out of the pipette and cylindrical ribbons formed. The extrusion step was conducted to obtain suitable size suppositories for the animals used in the laboratory scale experiment.

Example 2: Pharmacokinetics of Compound (1) and its Glucuronide Metabolite following rectal administration in male SD rats

For plasma samples: an aliquot of 30μΙ_ sample was added with 200μΙ_ IS (Propranolol, 40ng/rml_) in ACN. The mixture was vortexed for 5 min and centrifuged at 5800 rpm for 10 min. The 5 μΙ_ mixture was injected into LC-MS/MS.

For brain, rectum samples: tissue, taken post perfusion with saline, was homogenized for 2 min with 3 volumes (v/w) of homogenizing solution (PBS, pH 7.4). An aliquot of 30μΙ_ sample was added with 200 μΙ_ IS (Propranolol, 40 ng/mL) in ACN. The mixture was vortexed for 5 min and centrifuged at 5800 rpm for 1 0 min. The 5μΙ_ mixture was injected into LC-MS/MS. For faeces samples: tissue was homogenized for 2 min with 5 volumes (v/w) of homogenizing solution (MeOH/H₂0=1 :1 ). An aliquot of 30 μΙ_ sample was added with 200 μΙ_ IS (Propranolol, 40ng/ml_) in ACN. The mixture was vortexed for 5 min and centrifuged at 5800 rpm for 10 min. The 5μΙ_ mixture was injected into LC-MS/MS.

For 10-times diluted samples: An aliquot of 6 μΙ_ sample was added with 54μΙ_ blank to obtain the diluted samples, and the sample dilution factor is 10. The extraction procedure for diluted samples was the same as those for non-diluted samples. For 100-times diluted samples: An aliquot of 10μΙ_ sample was added with 90μΙ_ blank to obtain the diluted samples, and the sample dilution factor is 10. Then take 6μΙ_ sample, added with 54μΙ_ blank to obtain the diluted samples, and the sample dilution factor is 100. The extraction procedure for diluted samples was the same as those for non-diluted samples.

Results

Table 1 : Individual and mean plasma concentration-time data of Compound (1) after rectal dose of 100 mg/kg (API) in male SD rats

Table 2: Individual and mean brain concentration-time data of NYX- 205 after rectal dose of 100 mg/kg (API) in male SD rats

Sample contaminated during dissection

Table 3: Individual and mean rectum concentration-time data of Compound (1) after rectal administration of 100 mg/kg (API) in male SD rats

Table 4: Compound (1) concentration data in faeces after rectal administration of 100 mg/kg (API) in male SD rats

CV

rectal: 100 mg/kg (API) #19 #20 #21 Mean SD

(%)

Time Body weight

2670 2890 2700 2753 1 19 4.33 (hr) (g)

Dose (mg) 267 289 270 275 1 1 .9 4.33

(0-24) Cone, (ng/g) 245000 136000 66600 149200 89930 60.3

Volume (g) 4.55 3.72 4.56 4.28 0.482 1 1 .3

Mass (mg) 1 .1 1 0.506 0.304 0.641 0.422 65.8

0-24 hr Mass (mg) 1 .1 1 0.506 0.304 0.641 0.422 65.8 faeces fraction(%) 0.418 0.175 0.1 12 0.235 0.161 68.6 Table 5: Individual and mean plasma concentration-time data of glucuronides (metabolite) after rectal administration of Compound (1) at 100 mg/kg (API) in male SD rats

It has surprisingly been found that a compound of general formula (I), (II), (III), (IV) and/or (V), preferably wherein the compound is compound (1 ), may be delivered to brain tissue wherein the compound is administered as a rectal formulation.

Example 3: Kinase screen

A kinase screen was conducted in relation to Compound (1 ) (Eurofins KinaseProfiler™ kinase assay protocol v64). Table 6: IRAK4 results from kinase screen of Compound (1)

The results of the kinase screen indicate that lnterleukin-1 Receptor Associated Kinase 4 (IRAK4) is a target for Compound (1 ). lnterleukin-1 receptor-associated kinase-4 (IRAK4) is a serine-threonine kinase that participates as a key mediator in the interleukin-1 /Toll-like receptor (IL-1 /TLR) signaling cascades. Signaling through ILR/TLR results in the activation of the transcription factors kappa light chain enhancer of activated B cells (NF-kB) and activator protein-1 , ultimately inducing the generation of inflammatory cytokines. This data supports that Compound (1 ) is an anti-inflammatory compound.

Example 4: Cytokine array

Mouse macrophage Raw264.7 cells were plated. After 24 hours the cells were treated with either 20 μΜ Compound (1 ) or DMSO control. Thirty minutes later, cells were stimulated with either 10 ng/mL LPS or media added as control. Twenty hours later, media was collected and centrifuged at 14 OOOg for 5 mins. The supernatant was used in the mouse XL cytokine array (R&D Systems). Results

Table 7: Cytokine array data

LPS +

Analyte Control LPS Cmpd (1) Cmpd (1) LPS/LPS+Cmpd(l)

Reference spot 1.157168 1.374981 1.273044 1.15787

PCSK9 -0.00017 0.002856 0.000804 0.000404 7.077005

Leptin 0.000773 0.001928 0.000719 0.000395 4.879760

IL-la -0.000282 0.001062 0.000581 0.000303 3.50545

T Fa 0.001223 0.035554 0.001809 0.012457 2.854211

MMP-9 0.000379 0.016434 0.002402 0.006543 2.511684

CCL22/MDC 0.00746 0.013657 0.002472 0.006029 2.265077

CCL2/JE/MCP-1 0.258084 0.843733 0.18882 0.37413 2.25519

A compound of general formula (I), (II), (III), (IV) and/or (V), preferably compound (1 ), may be advantageously used to treat or minimise inflammation of the CNS, or a disease associated with inflammation of the CNS. Preferably, a compound of general formula (I), (II), (III), (IV) and/or (V), more preferably compound (1 ), inhibits one or more pro-inflammatory mediators including, but not limited to, kinases and cytokines. More preferably, the pro-inflammatory mediator is selected from the group consisting of: IRAK4, PCSK9, leptin, IL-1 a, TNFa, MMP-9, CCL22, CCL2, Syk, thromboxane A₂, and a combination thereof. In a particularly preferred embodiment, the pro-inflammatory mediator is IRAK4.

Claims

1 . A method for treating or minimising neuro-inflammation of the CNS in an individual requiring said treatment including:

(I) wherein D is halo;

Ri is H , C-i-10 alkyl, aryl, arylalkyl or R_ACO where RA is Ci-io alkyl or an amino acid;

R₂ is H, OH or OR₆;

wherein R₄ is H, CORD where RD is H , OH , Cno alkyl or an amino acid, CO2RC where Rc is Cno alkyl, CORE where R_E is H, C ₀ alkyl or an amino acid, COOH, COR_C where Rc is as previously defined, or CON H R_E where RE is as previously defined;

R₅ is H, CO₂Rc where Rc is as previously defined, or CORCORE where Rc and RE are as previously defined, and where the two R5 groups are attached to the same group they are the same or different, or

R6 is Cno alkyl, aryl or arylalkyl;

X is O, N or S;

Y is

2. A method for treating or minimising neuro-inflammation in an individual requiring said treatment including :

(ii) administering a therapeutically effective amount of a compound of general formula (I I) to the individual:

wherein

D is halo;

A and B together with the atoms between them form the group:

wherein

X is O, N or S;

Y is

where R₇ is H , or C- O alkyl; and

3. A method for treating or minimising neuro-inflammation in an individual requiring said treatment including: (i) providing an individual for whom treatment or minimisation of neuro- inflammation is required;

(ii) administering a therapeutically effective amount of a compound of general formula (III) to the individual:

wherein

D is halo; represents either a single bond or a double bond; Ri is H , C-i-₁₀ alkyl, aryl, arylalkyl or R_ACO where R_A is C-i-io alkyl or an amino acid;

Rs is O when ==is a double bond, or R5 is hydrogen or hydroxyl when ===== is a single bond;

Y is

where R_7A and R_7B are each independently H, OH or OR₇; with the proviso that at least one of R_7A and R₇B is OH or OR₇; and wherein R₇ is C₁-₁₀ alkyl, aryl or arylalkyl; wherein said administration minimises or treats neuro-inflammation in the individual.

4. A method for treating or minimising neuro-inflammation in an individual requiring said treatment including:

(IV) wherein

5. A method for treating or minimising neuro-inflammation in an individual requiring said treatment including:

(V) wherein D is halo, and wherein said administration minimises or treats neuro-inflammation in the individual.

6. A method of treating an individual having an IRAK4 associated condition or disease of the CNS, including administering to the individual a pharmaceutical composition including a compound of general formula (I) to the individual

(I) wherein

D is halo;

Ri is H, Cno alkyl, aryl, arylalkyl or R_ACO where RA is Ci-io alkyl or an amino acid;

R₂ is H, OH or OR₆;

R₃ is H, OH, or R_B where R_B is an amino acid or COR_A where R_A is as previously defined;

wherein R is H, CORD where RD is H, OH, Cno alkyl or an amino acid, CO₂Rc where Rc is Cno alkyl, CORE where RE is H, Cno alkyl or an amino acid, COOH, CORc where Rc is as previously defined, or CONHR_E where RE is as previously defined;

R₅ is H, CO₂Rc where Rc is as previously defined, or CORCORE where Rc and RE are as previously defined, and where the two R₅ groups are attached to the same group they are the same or different, or

R₆ is Cno alkyl, aryl or arylalkyl;

X is O, N or S;

Y is

where R₇A and R₇B are each independently H , OH or OR₇; with the proviso that at least one of R₇A and R₇B is OH or OR₇; wherein R₇ is C1-10 alkyl, aryl or arylalkyl; and

" == " represents either a single bond or a double bond.

7. A method of treating an individual having an IRAK4 associated condition or disease of the CNS, including administering to the individual a pharmaceutical composition including a compound of general formula (II)

(II) wherein D is halo;

Ri is H, or R_ACO where RA is CM O alkyl or an amino acid;

R₂ is H, OH, or R_B where RB is an amino acid or CORA where RA is as previously defined;

A and B together with the atoms between them form the group:

wherein

R₄ is H, CORD where RD is H , OH , C_no alkyl or an amino acid, CO₂Rc where Rc is C-i -10 alkyl, CORE where RE is H, Cno alkyl or an amino acid, COOH, CORc where Rc is as previously defined, or CON H RE where RE is as previously defined;

X is O, N or S;

Y is

where R₇ is H, or CM O alkyl; and

" represents either a single bond or a double bond.

8. A method of treating an individual having an IRAK4 associated condition or disease of the CNS, including administering to the individual a pharmaceutical composition including a compound of general formula (III)

(III) wherein D is halo;

" ===== " represents either a single bond or a double bond;

Ri is H , Cno alkyl, aryl, arylalkyl or R_ACO where RA is C₁-₁0 alkyl or an amino acid;

R₅ is O when ==is a double bond, or R₅ is hydrogen or hydroxyl when is a single bond;

Y is

where R_7A and R_7B are each independently H, OH or OR₇; with the proviso that at least one of R₇A and R₇B is OH or OR₇; and wherein R₇ is C₁-₁0 alkyl, aryl or arylalkyl.

9. A method of treating an individual having an IRAK4 associated condition or disease of the CNS, including administering to the individual a pharmaceutical composition including a compound of general formula (IV)

wherein D is halo.

1 0. A method of treating an individual having an IRAK4 associated condition or disease of the CNS, including administering to the individual a pharmaceutical composition including a compound of general formula (V)

(V) wherein D is halo.

1 1 . The method of any one of the preceding claims, wherein the compound i pound 1 as described herein, with the structure

1 2. The method of any one of the preceding claims wherein the individual has ischaemia and/or reperfusion injury to the CNS.

1 3. The method of any one of claims 1 to 1 1 , wherein the individual has neurodegenerative disease.

14. The method of claim 13, wherein the neurodegenerative disease is

Alzheimer's disease.

1 5. The method of any one of the preceding claims wherein the compound is administered rectally.

1 6. The method of any one of the preceding claims, wherein the compound inhibits one or more pro-inflammatory mediators selected from the group consisting of:

IRAK4, PCSK9, leptin, IL-1 a, TNFa, MMP-9, CCL22, CCL2, Syk, thromboxane A₂, and a combination thereof.

7. The method of claim 16, wherein the pro-inflammatory mediator is IRAK4.