Login| Sign Up| Help| Contact|

Patent Searching and Data


Title:
METHOD FOR PREVENTING AND/OR TREATING CHRONIC TRAUMATIC ENCEPHALOPATHY-I
Document Type and Number:
WIPO Patent Application WO/2015/000032
Kind Code:
A1
Abstract:
The present invention relates to a method of preventing and/or treating chronic traumatic encephalopathy.

Inventors:
VINK ROBERT (AU)
Application Number:
PCT/AU2014/050107
Publication Date:
January 08, 2015
Filing Date:
July 02, 2014
Export Citation:
Click for automatic bibliography generation   Help
Assignee:
EUSTRALIS PHARMACEUTICALS LTD TRADING AS PRESSURA NEURO (AZ)
International Classes:
A61K31/00; A61K31/48; A61K31/495; A61P25/00
Domestic Patent References:
WO2009009829A12009-01-22
WO2008067610A12008-06-12
Foreign References:
CA2831054A12013-12-26
Other References:
See also references of EP 3016642A4
Attorney, Agent or Firm:
DAVIES COLLISON CAVE (Melbourne, Victoria 3000, AU)
Download PDF:
Claims:
THE CLAIMS DEFINING THE INVENTION ARE AS FOLLOWS:

1 , A method of preventing and/or treating chronic traumatic encephalopathy, the method including administering to the subject an effective amount of a substance P receptor antagonist,

2. A method according to claim 1 , wherein the disease, condition or state is due to tali hyperphosphoryiation associated with chronic traumatic encephalopathy.

3. A, method according to claims 1 or 2, wherein the substance P receptor antagonist is a N 1 receptor antagonist, a NK2 receptor antagonist, or a N 3 receptor antagonist.

4. A method accordin to claim 3» wherein the NKl receptor antagonist is selected from one or more of the group consistin of Casopitant, CGP4 823, CP- 122,721, CP- 96,345, CP-99,994, F 888, QTi 8233.4, GR 94800, GR203040, GR-20S 171, GSK l 144814, GSK206-136, GSK424887, GW679769, HSP-1 17, L 703,606, L 732,138, L 733,060, L 742,694, L668.169, LY 303241 , LY 30 §70, LY 306740, Maropitant, MEN 1 1 149, Orvepilant, PD 154075, R-544, RP-67S8Q, RPR 100893, SCH619734, Spantide 11, Spantide III, Spendide, S 140333, Vestipitant, WIN-41,708, WIN-62,577, or a derivative, a variant, an analogue, a pharmaceutically acceptable salt, a tautomer or pro-drug thereof,

5. A method according to claim 3, wherein the N 2 receptor antagonist is selected from one or more of the group consisting of SR-48968, L-659877, GRl 03537, MGN- 10627, SR 14 190 and GR94800, or a derivative, a variant, an analogue, a pharmaceutically acceptable salt, a tautomer or a pro-drug thereof

6. A method according to claim 3, wherein the N 3 receptor antagonist is selected from one or more of the group consisting of SR- 143,801 , R82G, R486, SB22220 , L758,298 and NKP608, or a derivative^ a variant, an analogue, a pharmaceutically acceptable salt, a tautomer or a pro-drug thereof.

7. A method according to any one of claims 1 to 6, wherein the substance P receptor antagonist is administered Co the subject at a dose of 0,25 rug kg to 25 rng/kg,

8. A method according' to any one of claims 1 to 7, wherein the method reduces progression of the disease, condition or state associated with chronic traumatic encephalopathy.

9. Use of a substance P recepto antagonist in the preparation of a medicament for preventing and/or treating a disease, condition or state associated with .chronic traumatic encephalopathy.

! 0. A pharmaceutical composition when used to prevent and/or treat a disease, condition or state associated with chronic traumatic encephaiopathy, the composition including a substance P receptor antagonist.

1 1. A pharmaceutical composition according to claim 1.0, wherein the substance P receptor antagonist is a NKl receptor antagonist, a NK2 receptor antagonist, or a NK3 receptor an ta gon i st ,

12. A pharmaceutical, composition according to claim 1 1, wherein the NKl receptor aniagonist is selected from one or more of the group consisting Casopitant, CGP49823, CP- 122,721, CP-96,345, CP-99,994, F 888, GR 82334, GR 94800» GR20304Q, GR~ 205171, GSKI 144814, GSK206136, GSK424887, GW679769, HSP-117, L 703,606, L 732,138, L 733,060, L 742,694, 1.668,169, LY 303241 , LY 303870, LY 30674Q, Maropitant, MEN 1 1 149, Orvepitant, PD 154075, R-544, RP-67580, RPR 100893, SCH61. 734, Spantide 11, Spantide 111, Spendide, SR14.03.33., Vestipitant, WIN-41 J08, WIN-62,577, or a. derivative, a variant, an analogue, a pharmaceutically acceptable salt, a tautomer or a pro-drug thereof 13, A pharmaceutical composition according to claim 1 1 , wherein the N 2 receptor antagonis is selected from, one or more of the grou consisting of SR-48968, L-659877, OR 1.03537, MGN-10627, SRI 4190 and GR94800, or a derivative, a variant, an analogue, a pharmaceutically acceptable salt, a tautomer or a pro-drug thereof.

14, A pharmaceutical composition according to claim Π, wherein the N 3 receptor antagonist is selected from one or more of the group consisting of SR-143,801 , R820, R486, SB222200, L758,298 and NKP608, or a derivative, a variant, an analogue, a pharmaceutically acceptable sail, a tautomer or a pro-drug thereof,

15, A pharmaceutical composition according to any one of claims 9 to 14, wherein the pharmaceutical composition is used for either or both of:

i) preventing and/or treating a disease, condition or state associated with chronic traumatic encephalopathy;

ii) inhibiting progression of a disease, condition or state associated with chronic traumatic encephalopathy.

Description:
METHOD FOR PRE VENTING AND/OR TREATING CHRONIC

TRAUMATIC ENCEPHALOPATHY - 1

Field oTthe Invention

The present invention relates to a method of preventing and/or treating chronic traumatic encephalopathy.

Background of the Invention

Concussion has become an important public health problem in the United States, Australia and elsewhere internationally. It is common in a number of contact sports including the Australian football codes such as AFL and NRL, ice hockey, American football, and boxing, amongs others. In the United States alone, over 300,000 sports related concussions occur annually and numbers are increasing worldwide (Ell.enbogen et aL 2010, World Neurosurg. 74, 560-575). Concussive injuries are also a problem in the military and industrial worksites. In the case of the former, concussive injury resulting from exposure to the force of a detonation trigger similar neuropathological mechanisms leading to neuropatholog and sequelae indistinguishable to chronic traumatic encephalopathy (Goldstein et al (2012) Sei. Transl. Med. 4(134): 1 -16). Concussion causes no gross pathology, such as hemorrhage, and no abnormalities on structural brain imaging (MoCrory et al, 2009, Phys. Sportsmed. 37, 141 -159). There also may be no loss of consciousness, but many other complaints such as dizziness, nausea,, reduced attention and concentration, memory problems, and headache have been reported. A greater likelihood of unconsciousness occurs with more severe concussions. These types of concussive head impacts are very frequent in American football whose athletes, espeeially linemen ana linebackers, may be exposed to more than 1 ,000 impacts per season (Crisco et al., 2010, J. Athl.. Train. 45, 549-559), The effects of multiple concussions are becoming better recognized in these professional footballers, but much less is known about the long term-effects of repeated, concussion in the brains of amateur teenagers and adolescents, Moreover, the amateur codes of football are: less regulated than the professional codes, and the adolescent brain may be more vulnerable to concussion. The better-developed neck musculature of the professional footballer, the more strictly controlied tackling and the better aftercare of the concussed professional means that the long-term public health problem of concussion in sport is grossly underestimated.

Militar personnel who have experienced concussion experience a range of detrimental and chronic medical conditions. Concussion occurring amon soldiers deployed in Iraq is strongly associated with PTSD and physical health problems 3 to 4 months after the soldier return home, PTSD and depression are important mediators of the relationship between mild traumatic brain injury and physical health problems, PTSD was strongly associated with mild traumatic brain injury. It was reported that overall, 43,9% of soldiers who reported loss of consciousness met the criteria for PTSD, as compared with 27,3% of those with altered mental status, 36.2% of those with other injuries, and 9.1% of those with no injuries (Hoge et al, N Engl J Med, 2008; 358,453-63), Also, more than 1 in 3 returning military troops who have sustained a deployment-related concussion have headaches that meet criteria for posttraumatic headache (Theejer et al,, 201 0, Headache: J Head and Face. Pain 50, 1262-1272). It has been shown that nearly 15% of combat personnel sustained concussion whilst on duty (Hoge et al, N Engl J Med. 2008; 358,453- 63). Repeated concussion is a serious issue for combat personnel, with a study showing thai a majority of concussion incidents were blast related. The median time between events was 40 days, with 20% experiencing a second event within 2 weeks of the first and 87% within 3 months (MacGregor et al, 201 1 , J Rehab Research and Develop, 48, 1269-1278). Tile impact of concussion and PTSD has resulted in a significant economic burden, (The Congress of the United States - Congressional Budget Office, The Veterans Health Administration's Treatment of PTSD and Traumatic Brain Injury Among Recent Combat Veterans, February 2012).

While an isolated concussion has been widely considered to be an innocuous event, recent studies (Mc ee et al,, 2009, J Neuropath Exp Neurol 68, 709-735; Blennow et aL 2012, Neuron 76, 886-99) have suggested that repeated concussion is associated with the development of a neurodegenerative disorder known as chronic traumatic encephalopathy (CTE). CTE is regarded as a disorder that often occurs in midlife, years or decades after the sports or military career has ended (McKee et at, 2009, J Neuropath Exp Neurol 68, 709-735; Stem et al„ 2011 , Physical Med. Rehab. 3, S460-7), About one-third of CTE cases are progressive, but clinical progression is not always sequential or predictable. The clinical symptoms vary extensively, which is probably due to varying, multiple damage sites amongst athletes with the condition (Stern et al,, 201 1, Physical Med, Rehab. 3, S460- 7). The severity varies from mild complaints to severe deficits accompanied by dementia, Parkinson-like symptoms, arid behavioral changes. Clinical symptomB include neurological and cognitive complaints together with psychiatric and behavioral disturbances. Early neurological symptoms may include speec problems and impaired balance, while later symptoms include ataxia, spasticity, impaired coordination, and extrapyramidal symptoms, with slowness of movements and tremor .(Blennow et al. s 2012, Neuron 76, 886-99; Stem et al., 201 1, Physical Med. Rehab. 3, S460-7). Cognitive problems, such as attention deficits and memory disturbances, often become major factors in later stages of the disease, although may occur at varying times throughout the course of CTE, Psychiatric and behavioral problems include lack of insight and judgment, depression, disinhibition and euphoria, hypomania, irritability, aggressiveness and suicidal tendencies.

In post-mortem studies of athletes with CTE, the extensive, presence of neurofibrillary tangles has been reported (McKee et al., 2009, J Neuropath Exp Neurol 68, 709-735; Stern et al.., 2011, Physical Med. Rehab. 3, S460-7). Tangles are found tntracellularly in the cytoplasm of neurons and consist of threadlike aggregates of hyperphosphoryiated tau protein. Tau is a normal axonal protein that binds to microtubules via their microtubule binding domains, thus promoting microtubule assembly and stability. The hyperphosphoryiated form of tau causes disassembly of microtubules and thus impaired axonal transport, leading to compromised neuronal and synaptic function, increased propensity of tau aggregation, and subsequent: formation of insoluble fibrils and tangles. Unlike in Alzheimer's disease, tangles in athletes with CTB tend to accumulate perivascular! y within the superficial neocortical layers, particularly at the base of the sulci. Tau patholog in CTE is also patchy and irregularly distributed, possibly related to tire many different directions of mechanical force induced by physical trauma (McKee et al., 2009, J Neuropath Exp Neurol 68, 709-735), It is the accumulation of hyperphosphorylaied tau protein that is thought to result in the development of CTE and its associated psychiatric and behavioral disturbances.

Given these psychiatric and behavioral disturbances in athletes with CTE, there is a clear need for a therapeutic intervention to prevent and/or treat chronic traumatic encephalopathy,

A reference herein to a patent document or other matter which is given as prior art is not to be taken as an admission that that document or matter was known or that the information i contains was part of the common general knowledge as at the priority date of any of the claims.

Summary of the Invention

The present invention arises from studies into the association of substance P with tau hyperphosphorylation and development of chronic traumatic encephalopathy o a related condition having overlapping neuropathology and sequelae after concussive injury.

It is taught herein that the level of substance P is significantly increased foi lowing concussion. Substance P initiates tau hyperphosphorylation. It is prepared herein that substance P receptor antagonists are effective in reducing tau hyperphosphorylation thereby ameliorating the effects of CTE and related conditions.

Accordingly, in one aspect the present invention provides a method of preventing and/or treating chronic traumatic encephalopathy or a related condition i a subject, the method including administering to the subject an effective amount of a substance P receptor antagonist. In another aspect, the present invention also provides use o a substance P receptor antagonist in the preparation: of a medicament for preventing and/or treating chronic traumatic encephalopathy or a related condition in a subject. in a further aspect the present invention also provides a pharmaceutical composition when used to treat chronic traumatic encephalopathy or a related condition, the composition including a substance P receptor antagonist.

In still a further aspect the present invention also provides a method of inhibiting progression of a disease, condition or state associated with tau hyperphosphorylation in a subject, the method including administering to the subject an effective amount of a substance P receptor antagonist.

In still a further aspect the present invention also provides use of a substance P receptor antagonist in the preparation, of a medicament for inhibiting progression of disease, condition or state associated with tau hyperphosphorylation in a subject, for instance a concussive injury.

In a further aspect the inventio provides a method for treating a subject with a concussi ve injury, including: the ste of administering to said subject an effective amount of a substance P recepto antagonist. in a further aspect the invention provides methods for treating psychiatric and behavioural problems associated with CTE in a subject in need thereof, including the step of administering to said subject an effective amount of a substance P receptor antagonist.

In an embodiment the psychiatric and behavioural problems are selected from the group consisting of depression, irritability, disinhibition and euphoria, hypomania, aggressiveness and suicidal tendencies. In a further aspect the invention provides methods for treating cognitive problems associated with CTE, in a subject in need thereof, including the step of administering to said subject an effective amount of a substance P receptor antagonist.

In an embodiment the cognitive problems associated with CTE are selected from the group consisting of attention deficits and memory distrubanees.

Various terms that will be used throughout the specification have meanings that will be well understood by a skilled addressee. However, for ease of reference, some of these terms will now be defined.

The term "chronic traumatic encephalopathy (CTB). M as used throughout the specification is a condition appearing in response to repeated concussion resulting in accumulation of neurofibrillary tangles consisting of hyperphasp roi-ylated tau protein. The perivascular appearance of these neurofibrillary tangles within the superficial neocortical layers, and particularly at the base of the sulci, is unique to athletes and has been associated with the subsequent development of psychiatric and behavi oral disturbances.

The term "tau byperphosphorylation" as used throughout the specification is to be understood to mean the phosphorylated form of tau that causes disassembly of microtubules and thus impaired a onal transport, leading to compromised neuronal and synaptic function, increased propensity of tau aggregation, and subsequent formation of insoluble fibrils and tangles.

In this regard, a disease condition or state known as chronic traumatic encephalopathy is associated with accumulation of hyperphosp orykted tau protein, leading to compromised neuronal and synaptic function, increased propensity of tail aggregation, subsequent formation of insoluble fibrils and tangles, and the development of psychiatric and behavioral disturbances.

A related condition is a condition having overlapping neuropathology and sequelae. The term "'substance P receptor antagonist" as used throughout the specification is to be understood to mean an agent that directl or indirectly inhibits the binding of substance P to one of its receptors ' . In this regard, it will be appreciated that a substance F receptor antagonist includes a derivative, a variant, an analogue, a pharmaceutically acceptable salt, a solvate, a tautomer or a pro-drug of a substance P receptor antagonist, in this regard, substance P is an excitatory neurotransmitter and is a peptide having the structure RPKPEEFFGLM-NI¾ (SEQ ID NO 1), Methods for determining the ability of an agent to act as a substance P receptor antagonist are known in the art (Cascieri et al., 1983, J Biol Chem 258, 5158).

The term "variant" as used throughout the specification is to be understood to mean an amino acid sequence of a polypeptide or protein that is altered by one. or more anii.no acids. The variant ma have "conservative" changes, wherein a substituted amino acid has similar structural or chemical properties to the replaced amino acid (e.g., replacement of leucine with isoleucine). A variant may also have- "aotveonservative" changes (e.g., replacement of a glycine with a tryptophan) or a deletion and/or insertion of one or more amino acids. The term also includes within its scope any insertions/deletions of amino acids for a particular polypeptide or protein. A "functional variant" will be understood to mean a variant that retains the functional capacity of a reference protein or polypeptide.

Conservative substitutions typically include substitutions within the following groups: glycine and alanine; valine, isoleucine, and leucine; aspartie acid and glutamic acid; asparagine and glutamine; serine and threonine; lysine and argmine; and phenylalanine and tyrosine, Under some circumstances, substitutions, within the aliphatic group alanine, valine, leucine and isoleucine are also considered as conservative. Sometimes substitution of glycine for one of these can also be considered conservative. Other conservative interchanges include those within the aliphatic group aspartate and glutamate; within the amide grou asparagine and glutarnine; within the hydroxyl group serine and threonine; within the aromatic group phenylalanine, tyrosine and tryptophan; within the basic group lysine, argmine and histidine; and within the sulfur-containing group methionine and cysteine. Sometimes substitution within the group methionine and leucine can also be considered conservative. Substitutions as described above are contemplated within the scope of the present invention.

The term "prevent" as used throughout the specification is to be understood to mean an intervention that prevents or delays the onset: of a disease, condition or state in a. subject. The term "treat" as used throughout the specification is to be understood to mean an intervention that improves the prognosis and/or state of a subject with respect to a disease, condition or state.

The term "subject" as used throughout the specification is to be understood to mean human or animal subject,

The present invention furthermore has military applications such as administering: a substance P antagonist at an aid station shortly after a blast injury or traumatic events involving the head o during post recovery. it will also be understood that the present invention further includes within its scope veterinary applications. For example, the animal subject may be a mammal, a primate, a livestock animal (eg. a horse, a cow, a sheep, a pig, or a goat), a companion animal (eg. a dog, a cat), a laboratory test animal (eg. a mouse, a rat, a guinea pig, a bird, a rabbit), an animal of veterinary significance, or an animal of economic significance.

Brief Description of the. Figures

Figure 1 shows imnumohistoiogy using antibody for phosphoryla ed tau of sections of a human brain diagnosed with CTE demonstrating the perivascular appearance (A) of hyperphosphofyiated tau within the superficial neocoriical layers, and particularly at the base of the sulci ( ) (from Mc ee et al, 2009. J Neuropath Exp Neurol 68, 709-735). Figure 2 shows substance P imiriunoreaetivity in normal (A) and injured rats (B) at 3 days following mild concussive injury (B). In the injured animals, note the darkly stained, perivascular substance P positive axons and the presence of substance P positive neurones after injury (B) Bar - 1 OOnm.

Figure 3 shows the effects of a NK1 receptor antagonist (n-acetyl-L-tryptophan) on tau phosphorylation after concussive injury. Note that coneossive injury in the ra causes extensive tau phosphorylation (C,D) by 3 days after the conclusive event compared to non- injured animals (A,B), The administration of an i receptor antagonist (n-acetyl-L- tryptophan) at 30 min after the induction of injury results in almost complete inhibitio of tau phosphorylation at this 3 day time point (E,F).

Figure 4 shows a schematic model of ho w concussive events result in substance P release and subsequent hyperphosphorylation of tau, Neuronal sensor fibres surrounding blood vessels undergo stretch in response to a concussive event. The resultant mechanical stimulation activates mecha.noreceptors and triggers substance P release. Substance P binds to its receptor?, , activating an array of kinases known to be associated with hyperphosphorylation of tau. Hyperphosphorylation of tau destabilises microtubules and results in neurofibrillary tangles.

Figure 5 shows stress fields following simulated rotational acceleration in models replicating brain tissue with no sulci (A) to brain tissue with complex sulci formation (B- D). Higher stress is indicated as bl ack and is focused at the base of the sulci irrespective of the sulcus morphology.

General Description of the Invention

As described above, the present invention provides a method of preventing and/or treating chronic traumatic encephalopathy or a related condition in a subject, the method including administering to the subject an effective amount of a substance P receptor antagonist . This embodiment of the present invention is directed to preventing and/or treating a disease, condition or state associated with tau hyperphosphoryiation by administering to a subject one or more substance P receptor antagonists.

Tau hyperphosphoryiation may be induced by a variety of reasons, including for example, a concussive event or a mechanical impact that activates brain rneehanoreeeptors. In this regard, tau hyrjerphosphorylation may be associated, for example, with either or both an accumulation of hyperphosphoryiated tau over time as measured within the one subject, or may be an accumulation of hyperphosphoryiated tau in one subject compared to the accumulation of hyperphosphoryiated tan in a population.

Diseases, conditions or states associated with accumulatio of hyperphosphoryiated tau in subject in the various embodiments of the present invention include chronic traumatic encephalopathy (CTE) .

Chronic traumatic encephalopathy (CTE) is normally classified as a disease associated with accumulation of tangles containing hyperphosphoryiated tau, with these tangles tending to accumulate peri vascular] y within the superficial neocortical layers, particularly at the base of the sulci, There is currently no blood or laboratory test that is definitive for the diagnosis of CTE, with disease confirmation usually occurring after postaoretm examination of brain tissue, Nonetheless, a number of clinical criteria plus a history of concussive events in the subject are usually sufficient in making a tentative diagnosis. In this regard, the Diagnostic and Statistical Manual of Mental Disorders (American Psychiatric Association) is commonly used to assess a number of parameters to provide an indication of the presence and severit of CTE in a subject. Nuclear, medical imaging, including Positron Emission Tomography (PET), may also be used to assess the presence and severity of CTE, Methods of assessing CTE in a subject using PET include for example Small el el (2013) Am. . Geriatr. Psychiatry. 21; 138-144.

Accordingly, in a further embodiment the invention may include a CTE diagnostic step which may be performed by injecting the subject with a PET molecular imaging probe (to visualise CTE in living humans). Such imagin probes are known, for instance, FDDNP(2- 1 - {6-[(2 ' F~ 18|fluoroethyl)(methy l)aniinoJ-2*napthyi }ethy!idene)ma!ononitrite, Such probes are able to visualise tan tangles.

In a further embodiment the diagnostic step may include an assessment of the plasma levels of total tau (T-tau) using an immunoassay for instance, as described in Rissen et al, Nature Biotechnology 2010;28:595-599 (which is incorporated by reference in its entirety).

In an embodiment diagnosis of CTE may be made based on a plasma level of Tau (based on the aforementioned assay) of above 1.5 ngL "1 , for instance, above 1 ,6, above 1.7, above 1.8, above 1,9, above 2.0, above 2,1 , above 2,2, above 2,3, above 2,4, above 2,5, above 2,6, or above 2.7 ngL "1 .

In one specific embodiment, the disease, condition or state associated with accumulation of hyperphosphorylated tau is chronic traumatic encephalopathy.

In another specific embodiment, the diverse, condition or state associated with accumulation of hyperphosphorylated tau is a concussive event or injury.

Substance P binds to a number of receptors including the N .1 receptor (neurokinin 1 receptor), the N 2 receptor and the NK3 receptor. Substance P antagonists inhibit the binding of substance P to any one of its receptors. Accordingly, NK.1 , NK2, and NK3 receptor antagonists will also function as substance P receptor antagonists. It will be appreciated that the term "substance P" includes within its scope various variants, truncated forms or analogues of the peptide, for example as described in US Patent No, 4,481,139.

The identification of a substance as a substance P receptor antagonist may be determined by their ability to inhibit tire binding of substance P at its receptor sites in bovine caudate tissue, employing radioactive ligands to visualize the tachykinin receptors by means of autoradiography, The substance P antagonizing activity of the herein described compounds may be evaluated b using the standard assay procedure described by M.A. Cascieri et al., as reported in the Journal of Biological Chemistry, Vol, 258, p. 5158 (1983). The method involves determining the concentration of the individual compound required to reduce by 50% the amount of radiolabelled substance P Jigan s at their receptor sites in said isolated bovine thereby affording characteristic ICjo values for each compound tested.

Table 1 - NK1 Receptor Antagonists

Chemical Code Chemical Name

GSK424887 Unknown

OW679769 Unknown

HSP-I 17 3-Piperidinamine, N-[[2 ; 3-dihydro-S--(l -n ethylethyI)-7- benzofuranyl]roethy )2-phenyl ^ , dihydrochloride, (2S-cis)-

L 703,606 1 -Azabicycio [2.2.]octan-3-amine ( 2-(diphenylrnetbyl)-N-f(2- idophenyl)methylj-, (2S-eis)-, oxalate

L 732,138 N-acetyl-L-tryptophan

L 733,060 ((2S,S)-3-((3,5-bis(trinu0romethyl)phcnyl)rneihyloxy)-2-phen yl pSperidine

L 742,694 (2-(S)-(3,5-bis(trifluromet l)bpnzylGxy)-3- S)-ph.eny]-4-(5-(3-ox0- 1 ,2, 4-tria¾ot-o)meihylraorplioline

L668J69 L-Phenylalanine, N-[2-[3-[[N-[2-(3-[(N-[2-[3-atnino-2-oxo-l - pyrrolidiny])-4-methyl-l-oxopeni]yl]-L-methionyl-L-giiitamin y|-D- tryplophyl-N-methyl-L-phenylaIai7yl]an5ino]-2-oxo-l -pyrrolidinyj]- 4-melJiyl-l.-.oxopenlyl]-L-niethionyi-L''glutaminyl-D-trypto phyl-N- mcthyl-.cyclic (8-> l)-peptide ( [3 -[1 fS*[R*(S*)] . ] > 3R*]>

L Y 303241 1 -Pi ei'azineacctaraide, N-[2-[acety![(2- me ' thoxyphenyI)metbyl]amino]-l-(l H-indol-3-ylraethyl)(etliyl]-4- phenylr,. (R)-

LY 303870 (R)- 1 - [ -(2-methoxy benxyl)acety!amino] -3-( 1 H-indol-3-yl )-2-[N-(2~

(4-(pi pcridinyi)piperidm- 1 -yl)ac.etyl)amino jpropane

LY 306740 1 -Piperazineacetamide, N-[2- < acetyl[(2- methoxypehenyl)methyl]amino]-l-(lH-indol-3-y cyclohexyK (R)-

Maropitant <7R-8S)-N-[(5-letl-Bu.tyl-2-iMethoxyphenyl)niethyl]-7- [di(phenyI)metl yJ]- 1 -azahicyclo[2.2.2]octan-S-amine

MEN 11149 2-(2-naphthyl)-l -N[(l R,2S)-2-N-[l(H)indo.l-3- ylcarbonyljaminocyclohexanecarbonyl] - 1 -[NT -ethyl -N 5 -(4- methyiphen lacetyl)] di ami noethane

Orvcpitant (2R t 4S)-N-((R)~l-(3,5-bis(trifluow^

2-methylphenyl)-N-methyl-4-( S)-6-oxohexahydrQpyTrolo[ ' l,-2- a]pyrazin~2( 1 H)-yi)piperidint;- 1 -earboxamide maleate Chemical Code Chemical ame

PD 154075 (2-beiKofuran)-CH20CO] R)-alp fi-McT^-{S)-NHCH(CH3)Ph

R-544 Ac- Thr- D -Trp (FOR)- P he -N- eB zi

RP-67580 (3aR, 7aR)-7 -diplieiiyl-2[l-imino-2(2-meihoxyphenyl)- (cthyl]+++perhydroisoindot-4-one hydrochloride

RPR 100893 (3aS,4§ s 7aS)-7, 7^iphenyl-4 2-m thoxyphenyl)-2-[(S)-2-(2- methoxy phen l )pro:pri o ny 1 ]perh yd ro i so ind ol · -ο 1

SCH619734 (5S)~8(S [[1 (R)-[ ,5 blsltriflitororaeth S)phenyl]ethoxy]methyi]-8- phenyl- 1 ,7-dia?:aspiTOi4,5]decari-2"0ne)

Spantide II D-NicLys 1 > 3-Pal3, D-C12Phe5, Asn6, D-Trp7.0 ? Nlel 1 -substance P

Spantide HI L-Norleucinarnide, N6--(3-pyridinylcarbonyl)-D-lysyl-L-pro]yj-3-(3- pyridiny^i-alanyl-L-prolyl-S^-dichioiO-D-phenyialanyl-L- asparaginyl-D-iryptophyj-L-phenylalanyl-3-(3-pyrid[ny])-D-al an>4- L-leucyl-

Spendide Tyr-D-Phe-Ph.e-D-iiis-Leu- .et-NPI2

SRI 40333 {S)-l -[2-[3-(3,4-dichlorphenyl)- (3-jsopropoxyphenylacety!)

piperidin-3-yl] ethyl]-4~phenyJ-l azaniabicycJo 2,2.2]octane

Vesiipitant (2S)-N- (IR)-l-[3,5-bis(lritIuoromethyI)phenyl]elhyI]-2-(4-l]uoro-2- mcthy lphenyl)-N-methylpiperazine- 1 -carboxamide

WIN-41 ,708 (17 bela-hy droxy- 17al ha-ethy ny 1 - 5alpha- androstan ol 3,2 - b]pyrirnido[l,2-a]ben.zimidazole

WIN-62,577 1 H-Benzimidazo[2, 1 -b]eycl.openta] ' 5,6]naphtha[l ,2-gjqinnazolin- 1 - ol, 1 -ethynyl-2 ( 3 5 3a,3b,4,5 5 l 5, 15 a, 15b.16, 17, 17a-dodeachydro~ 1 5 a, 17a-dimethy (1R, 3aS, 3 bit, 15aR, 15bS, 17aS)~

Table 2 - 2 Receptor Antagonists

MEN 10627 Cy.d (M.ei-Asp-Trp-Phe-Dap-Leu3cyclo(2 eta'-5beta)

1 SR 144190 (R)-3-( l -[2r(4-b«n¾oyt-2-(3,4-difluorop ' henyl -raoipho-iin-2-yl)- ethy 1] ~4 -phe n yip iperid in- 4-y 1) - 1 -d i methylu rea

I GR 94800 PhCO-AIa-A ' la-D-Trp-Phe-D-Pro-Pfo-Nle-NH2

Table 3 - MO .Receptor Antagonists

Chemical Code Chemical Name

SR-142,801 (S)-(NH>(3-(I -benzo l^

yl)piOpyl)-4-phei'iylpipeiidin-4-y1)-N -methyl acetaide

R820 3-indoiylcarbonyl-Hyp-Phg-N( e)-BKl

R486 H-Asp-Ser-Phe-Trp-beta-Ala-Leu-Mel-NH:2

SB 222200 (S ).- (-) -N-(a-ethy lbenzy 1) -3 -methy 1-2-pheny i uino ii nc-4-carbox i mi de

L 758,298 Phosphonic acid, [3-[[2- l ^[3,5-bis(trfluoromethyl)phenyl]ethoxyl-3 - (4-fluorophenyl)-4-morp.holinyl]me.thyl]-2 ,5-dihydre-4oxo- 1 11- 1 ,2,4- triazol-l -yl]- [2R-[2a(R*), 3a]]-

ΝΚ-60» (2R,4S)-N-(l-[3.5-bi.sCtril1uorrae I)-benzoyl)-2-(4-chIoiO-ben2yl)- 4-piperidinyl]-quinoline-4-carb.oxamide

Other examples of substance P receptor antagonists are as described in US patents 4,481,139 and 5,977,104. Examples of NKl receptor antagonists are as described in OS patent 5,990,125.

In one embodiment, the substance P receptor antagonist is one or more of a NKl receptor antagonist, a NK.2 receptor antagonist, and a NK3 receptor antagonist.

In one embodiment, the NKl receptor antagonist is selected from one or more of the group consisting of Crepitant, CGP49823, CP- 122,721, CP-96,345, CP-99,994, FK. 888, GR 82334, GR 94800, GR203040, GR-20S17I, GS 1144814, GSK206136, GSK424887. GW679769, HSP-1 17, I, 703,606, L 732,138, L 733,060, L 742,694, L668.169, LY 303241 , LY 303870, LY 306740, Mafopitant, MEN 1 1 149, Orvepitant, PD 154075, R- 544, RP-67580, RPR 100893, SCH619734, Spantide II, Spantide 01, Spendide, SRI 40333, Ves ipitant, WIN-41,708, WIN-62,577, or a derivative, a variant, an analogue, a pharmaceutically acceptable salt, solvate, tautomer or pro-drug thereof.

In another embodiment, the NO receptor antagonist is selected from one or more of the group consisting of SR- 8968, L-659877, GRl 03537, MGN 0627, SR 1.44190 and GR94800, or a derivative, a variant, an analogue, a pharmaceutically acceptable salt, tautomer or pro-drug thereof.

In another embodiment, the N 3 receptor antagonist is selected from one or more of the group consisting of SR-143,801, R82Q, R486, SB222200, L758.298 and N P60S, or a derivative, a variant, an analogue, a pharmaceutically acceptable salt, tauiomer or rodrug thereof.

In one embodiment, the substance P receptor antagonist is L- 732.138, namely N-aeetyl-L- tryptophan, or a derivative, analogue, pharmaceutically acceptable sa t, solvate, tautomer or pro-drug thereof. Examples include lipid soluble analogues, N-acetyl-L-tryptophan 3,5- bis(triiluoromethyl)benzyl ester and N-ace.tyl tryptophan methyl ester.

One or more substance P receptor antagonists may also be used in the preparation of a medicament for preventing and/or treating chronic traumatic encephalopathy or a related condition.

Accordingly, in another embodiment the present invention provides use of a substance F receptor antagonist in the preparation o a medicament for preventing and/or treating chronic traumatic encephalopathy or a related condition. One or more substance P receptor antagonists may also be used in a pharmaceutical composition, to prevent and/or treat chronic traumatic encephalopathy or a related condition.

Accordingly, in another embodiment the present invention provides a pharmaceutical composition when used to prevent and/or treat chronic traumatic encephalopathy or a related condition, the composition including a substance P receptor antagonist.

The administration of one er more substance P receptor antagonists may also be used to inhibit progression of the disease, condition or state associated with chronic traumatic encephalopathy or a related condition in the subjeet.

Accordingly, in another embodiment the present invention provides a method of inhibiting progression of chronic; traumatic encephalopath or a related: condition, the method including administering to the subject an effective amount of a substance P receptor antagonist.

The effective amount of a substance P receptor antagonist to be delivered in the various embodiments of the present invention is not particularly limited, so lon as it is within such an amount and in such a form that generally exhibits a useful or therapeutic effect, The term ''effective amount" is the quantit which when delivered, improves the prognosis of the subjeet. T ' he amount to be delivered will depend on the particular characteristics of the condition being treated, the mode of delivery, and the characteristics of the subjeet, such as general health, other diseases, age, sex, genotype, bod weight and tolerance to drags.

In a embodiment, an effective amount of a substance P receptor antagonist is an amount to be delivered to restore plasma levels of total iau (T-tau) (for instance by the immunoassay identified hereinbefore) to less than I ngl, "1 , for instance, less than 0, ngL "1 , or less than 0.8 ngL "1 . This may involve a single, dose or repeated dosages. Accordingly, a suitable dosage of the substance P receptor antagonist for delivery to the desired si te of action in the various embodiments of the present invention may be selected, in an embodiment the method relates to a method for treating a concussive injury which involves a patient being exposed to multiple (more than one) concussive events, In such a method, the attendant physician would determine that the subject is concussed and that the subject has had at least one previous concussion. Methods for determining whether or nor a subject has been concussed includes for instance a variety of neuropsychological assessment tools (Kelly et ah, 2012, Arch Clin Meuropsycho 27, 375-88; Echemendia et al., 2012, Clin Neuropsycbol 26, 1077- 1), However, the detection of loss of memory, an alteration of mental state (mental cloudiness, headache, dizziness, confusion, disorientation), possible loss of consciousness, or focal neurological deficits is more commonly used for on-field diagnosis of a concussive event, Other diagnostic criteria are outlined in detail in the Ameriean Society for Sports Medicine position statement (Br I Sports Med, 2013, 47, 15-26) and are summarised on the regularly updated Centres for Disease Control and Prevention (USA) website (http: /wwv\ f ,cdc,gov concussion/ sports/index.hlml). Once this has been established the physician would then administered an effective amount of a substance P receptor antagonist.

In one embodiment, the dosage of the substance P receptor antagonist administered to a subject in. the various embodiments of the present is in the range from 0.1 nig/kg to 100 mg/kg.

In a specific embodiment, the substance P receptor antagonist is administered to the subject at a dose of 0.25 mg/kg to 25 mg/kg. For example, a suitable dose of N-acetyl- tryptophan is 2.5 mg/kg.

In. an embodiment the effective amount is an amount which is able to maintain the blood concentration of the substance P receptor antagonist in the therapeutic range for at least 3 days, for instance at least 4 days, at least 5 days, at least 6 days, at least 7 days, at. least 8 days, at least 9 days,: at least 10 days, at least 1 1 days, at least 12 days, at least 13 days, at least 14 days, at least 15 days, at least 16 days, at least 37 days, at least 18 days, at least 19 days, or at least 20 days.

In an embodiment the effective amount is administered as a single dose.

In an embodiment the effective amount is administered as a single oral dose.

Accordingly, in anothe aspect the invention provides a method for treating a subject which has been exposed to multiple concussive- events including the step of administering to the subject a substance P receptor antagonist as a single oral dose in an amount which is able to maintain the blood concentration of the substance P receptor antagonist in the therapeutic range for at least 3 days, wherein the administration step is performed after the second eoucussive event and again after each additional concussive event as required.

In an embodiment the substance P receptor antagonist is administered within 24 hours of the concussive event.

In an embodiment administration is provided within 20 hours such as within, 19 hours, 18 hours, 17 hours, 16 hours, 15 hours, 14 hours, 13 hours, 12 hours, 1 1 hours, 10 hours, 9 hours, 8 hours, 7 hours, 6 hours, 5 hours, 4 hours, 3 hours, 2 hours and within 1 hour, of the concussive event.

In an embodiment the oral dose is in the form of a tablet, capsule, drink solutions or parenteral.

Generally, the dosage of the substance P receptor antagonist in a pharmaceutical composition may be in the range from 10-5,000 rag per subject, and typicall will be in the range of 50-2,000 rng per subject.

Suitable dosages are generally as described in US patents 4,990,125 and US 5,977, 1.04 (incorporated within by reference in its entirety). Examples of formulations are described in US patent 5,990,1 5 (incorporated within by reference in its entirety).

Methods for the preparation of pharmaceutical compositions are known in the art, for example as described in Remington's Pharmaceutical Sciences, 18th ed., 1990, Mack Publishing Co,, Eastern, Pa. and U.S. Pharmacopeia: National Formulary, 1.984, Mack Publishing Company, Easton, Pa.

As discussed previously herein, administration and delivery of the compositions may be for example by the intravenous, intraperitoneal, subcutaneous, intramuscular, oral, or topical route, or by direct injection. The mode and route of administration in most cases will depend on the severity and frequency of the eoncussive events.

The dosage form, frequency and will depend on the mode and route of administration.

As described above, the administration of the substance P receptor antagonist and other agents may also include the use of one or more pharmaceutically acceptable additives, including pharmaceutically acceptable salts, amino acids, polypeptides, polymers, solvents, buffers, exeipients, preservatives and bulking agents, taking into consideration the particular physical, microbiological and chemical eharacteristics of the agents to be: administered.

For example, the substance P receptor antagonist and/or the other agents can be prepared into a variety of pharmaceutically acceptable compositions in the form of, e.g., an aqueous solution, an oily preparation, a fatty emulsion, an emulsion, a lyophilised powder for reconstitutton, etc. and can be administered as a sterile and pyrogen free intramuscular or subcutaneous injection or as injection to an organ, or as an embedded preparation or as a transmucosal preparation through nasal cavity, rectum, uterus,: vagina, lung, etc. The composition may be administered in the form of oral preparations (for example solid preparations such as tablets, eaplets, capsules, granules or powders; liquid preparations such as syrup, emulsions, dispersions or suspensions).

Compositions containing the substance P receptor antagonist and/or the other agents may also contain one or more pharmaceutically acceptable preservatives, buffering agents, diluents, stabilisers, chelating agents, viscosity enhancing agents, dispersing agents, pH controllers, of isotonic agents.

Examples of suitable preservatives are benzoic acid esters of para-hydroxybenzoic acid, propylene, glycol, phenols, phenylethyl alehohol or benzyl alcohol. Examples of suitable buffers are sodium phosphate salts, citric acid, tartaric acid and the like. Examples of suitable stabilisers are, antioxidants such as alpha-tocopherol acetate, ajpha-thioglycerin, sodium nietabisulphite, ascorbic acid, acetylcysteine, S-hydroxyquinoline, chelating agents such as disodium edetate. Examples of suitable viscosity enhancing agents, suspending or dispersing agents are substituted cellulose ethers, substituted cellulose esters, polyvinyl alehohol, polyvinylpyrrolidone, polyethylene glcols, cavbomer, poiyoxypropylene glycols, sorbitan monooleate, sorbitan sesquioleate, polyoxyethylene hydrogenated castor oil 60.

Examples of suitable pi I controllers include hydrochloric acid, sodium hydroxide and the like. Examples of suitable isotonic agents are glucose, D-sorbitol or D-mannitol, sodium chloride.

The administration of a substance P receptor antagonist and/or the other agents in the various embodiments of the present invention may also be in the form of a composition containing a pharmaceutically acceptable carrier, diluent, excipient, suspending agent, lubricating agent, adjuvant, vehicle, delivery system, emulsifier, disintegrant, absorbent, preservative,- suriaetant, colorant, glidant, anti-adherant, binder, flavorant or sweetener, taking into account the physical, chemical and microbiological properties of the agents being administered. For these purposes,- the composition may be administered orally, parenferally, by inhalation spray, adsorption, absorption, topically,: rectal I y, nasally, mucosaliy, Ira sdertn ally, bucally, vaginally, intraventricularly, via an implanted reservoir in dosage formulations containing conventional non-toxic pharmaeeutically-aeeepiahie carriers, or by any other convenient dosage form.- The term parenteral as used herein includes subcutaneous, intravenous, intramuscular, intraperitoneal, intrathecal, intraventrieuiar, intrasternal, and intracranial injection or infusion techniques.

When administered- parenteraily, the compositions will normally be in a un.it dosage, sterile, pyrogen tree injectable form (solution, suspension or emulsion, which may have been reconstituted prior to use), which is generally isotonic with the blood of the recipient with a pharmaceuticall acceptable carrier. Examples of such sterile injectable forms are sterile injectable aqueous or oleaginous suspensions. These suspensions may be formulated according to techniques known in the. art using suitable vehicles, dispersing or wetting agents and suspending agents. The sterile injectable forms may also be sterile injectable solutions or suspensions in non-toxic p¾renterally acceptable diluents or solvents, for example, as solution in l ,3~butanecliol. Among the pharmaceutically acceptable vehicles and solvents that may be employed are water, ethanol, glycerol, saline. Ringer's solution, dextrose solution, isotonic sodium chloride solution, and Hanks' solution. In addition, sterile, fixed oils are conventionally employed as solvents or suspending mediums. For this purpose, any bland, fixed oil may be employed including synthetic mono- or di-glyeerides, corn, cottonseed, peanut, and sesame oil. Fatty acids such as ethyl o!eate, isopropyJ myri state, and oleic acid and its gl-yceri.de derivatives, including olive oil and castor oil, especially in their polyoxyethylated versions, are useful in the preparation of injectables. These oil solutions or suspensions may also contain long-chain alcohol diluents or disper ants.

The carrier may contain minor amounts of additives, such as substances that enhance solubility, isotqnjcity, and chemical stability, for example anti-oxidanis, buffers and preservatives. In addition, the compositions may be in a form to be reconstituted prior to administration. Examples include lyophiiisation, spray drying and the like to produce a suitable solid form for reeonstiuiiion with a pharmaceutically acceptable solvent prior to administration.

Compositions may include one or more buffers, bulking agents, isotonic agents- and c-ryoprotectants and lyoprotectants. Examples of excipients include, phosphate salts, citric acid, nofl-yed cing such as sucrose or trehalose * polyliydroxy alcohols, amino acids, methylamiries,. and inotropic salts which are usually used instead of reducing sugars such as maltose or lactose.

When administered orally, the substance P receptor antagonist will usually be formulated into unit dosage forms such as tablets, caplets, cachets, powder, granules, .beads,, chewable lozenges, capsules, liquids, aqueous suspensions or solutions, or similar dosage Forms, using conventional equipment and techniques known in the art. Such formulations typically include a. solid, semisolid, or liquid carrier, Exemplary carriers include excipients such as lactose, dextrose, sucrose,: sorbitol, mannito.1, starches, gum acacia . , calcium phosphate, mineral oil, cocoa butter, oil of theotaroma, alginates, tragacanth, gelatin, syrup, substituted cellulose ethers, polyoxyethylene sorbitan monolauraie, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium siearate, and the like.

A tablet ma be made by compressing or molding the agent optionally with one or more accessory ingredients. Compressed tablets may be prepared b 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, inert diluent, surface active, or dispersing agent. Moulded tablets ma be made by moulding in a suitable machine, a. mixture of the powdered active ingredient find a suitable earner moistened with an inert liquid diluent,

The administration of the substance P receptor antagonist may also utilize controlled release technology. The substance P receptor antagonis ma also be administered as a sustained-release pharmaceutical composition. To further increase the sustained release effect, the agent may be formulated with additional components such as vegetable oil (for example soybean oil, sesame oil, camellia oil, castor oil, peanut oil, rape seed oil); middle fatty acid triglycerides; fatty acid esters such as ethyl oleate; polysiloxane derivatives; alternatively, water-soluble high molecular weight compounds such as hyaluronic acid or salts thereof earboxymcthylcellulose sodium hydiOxypropylceliulose ether, collagen polyethylene glycol polyethylene oxide, hydroxypropylmeihyleellujosemethylcellu!ose, polyvinyl alcohol, polyvinylpyrrolidone.

Alternatively, the substance ? receptor antagonist may be incorporated into a hydrophobic polymer matrix for controlled release over a period of days. The agent may then be moulded into a solid implant, or externally applied patch, suitable for providing efficacious concentrations of the agents over a prolonged period of time without the need for frequent re-dosing. Such controlled release films are well, known to the art, Other examples of polymers commonly employed for this purpose that may be used include nondegradable ethyiene-vmyl acetate copolymer a degradable lactic acid-glycolic acid copolymers, which may be used externally or internally. Certain hydrogels such as poly(hydroityethylmethacrylatc) or poly(vinylalcohol) also may be useful, but for shorter release cycles than the other polymer release systems, such as those mentioned above.

The earner may also be a solid biodegradable polymer or mixture of biodegradable polymers with appropriate timer-release characteristics and release kinetics. The agent may then be moulded into a solid implant suitable for providing efficacious concentrations of the agents over a prolonged period of time without the need for frequent re-dosing. The agent can he incorporated into the biodegradable polymer or polymer mixture in any suitable manner known to one of ordinary skill in the art and may form a homogeneous matrix with the biodegradable polymer, or may be encapsulated in some way within the polymer, or may be moulded into a solid implant. For topical administration, the substance P receptor antagonist may he in the form of a solution, spray, lotion, cream (for example a non-ionic cream), gel, paste or ointment Alternatively, the composition may be. delivered via a liposome, nanosome,- rivosome, or nuiri-difluser vehicle,

Tt will he appreciated that other forms of administration of agents are also contemplated, including the use of a nucleic acid encoding a polypeptide for delivering of such agents.

Description of Specific Embodiments

Reference will now be made to experiments that embody the above general principles of the present invention. However, it is to be understood that the following description is not to. limit the generality of the above description.

Example 1

Concussion results in accumulation of hyperphosphorylated tau,

A number of clinical and experimental studies have now shown that there is an accumulation of hyperphosphorylated tau followin coneussive injury. Accumulation of neurofibrillary tangles containing hyperphosphorylated tau is a hallmark pathology of chronic traumatic encephalopathy, especially when, this accumulation is perivascular and predominantly found within the superficial neocortical layers, particularly at the base of the sulci. In human studies ( c ' ee et al., 2009, J Neuropath Exp Neurol 68 . , 709-735} such a distribution of hyperphosphorylated tau is readily apparent in subjects who have a history of repeated concussive events (Figure 1). In this particular example, localization of hyperphosphorylated tau is shown in an NFL football player with a history of repeated concussion. Note the perivascular localization of hyperphosphorylated tau (A) with highest accumulations at the base of the sulci. This pathology is unique to chronic traumatic encephalopathy, Similar accumulations of hyperphosphorylated tau have been shown ■following- experimental concussion in animals, although the absence of sulci in the experimental animals used in these studies to date has precluded the demonstration that such .accumulation replicates the human pattern of localisation at the base of the sulci.

Example 2

Concussion remits in perivascular substance P release

Having established that hypeiphosphorylated tau accumulates perivascular!}' * following concussive injury, we used an animal model of concussion to investigate whether concussion causes perivascular release of substance P, We developed a rodent model of concussion to replicate the concussive event (Donkin et al., 2004, 7th International Neurotrauma Symposium, p 75-78, Medirnood Publishers, Bologna, Italy) and subsequently deteTmined whether substance P was released after such an event. There was a clear increase in brain perivascular substance P immunoreactivity alter the concussive event (Figure 2). We propose that mechanical stimulation of sensory nerve fibres was responsible for this peri vascular release of -substance P.- These results are consistent with previous studies in non-brain tissue demonstrating that mechanical stimulation of sensory nerve fibres induces substance P release (Aug et al., 2011 , PLoS One 6, e24535).

Example 3

Administration of a NKJ receptor antagonist prevents tau phosphorylation

Having shown that mechanical injury causes release of substance P- after. concussive injury, we then investigated whethe a substance P NK.1 receptor antagonist reduces tau hypcrphosphorlation after concussive injury. Figure shows the effects of an N 1 receptor antagonist (n-aeetyl-L-tryptophan) on tau phosphorylation after concussive injury. Note that concussive injury in the rat causes extensive tau phosphorylation (CD) by 3 days after the concussive event compared to non-injured animals- (A,B). The administration of the N 1 receptor antagonist at 30 mill after the induction of injury results in almost complete inhibition of tau phosphorylation at this 3 day time point (B,P). Thus, administration of an NKl receptor antagonist prevents tau hyperphosphoryiation and thus prevents the development of CTE,

Example 4 The mechanism by which substance P anlaganisis inhibit tau hyperphosphorylation Having established that substance P antagonists inhibit tan hyperphosphorylation after eoncussive injury, we propose that the mechanism by which this occurs is through substance P binding to its N 1 receptor and activating various biochemical en¾ymes (kinases) that will result in the hyperphosphorylation. of tau and the development of chronic traumatic encephalopathy (CTE). The mechanism of this activation is shown in Figure 4. Activation of mechanoreceptors during eoncussive injury causes the release of substance P (example 2 and Ang et al., 201 1 , PLoS One 6, e24535). Furthermore the extent of the substance P release and the activation of its receptors is proportional to the frequency of the stimulation (Matityh, 2002. J. Clin. Psychiatry 63, 6-10). Specifically, repeated exposure to a SP-releasin mechanical stimulus such as concussion increases expression of the substance P receptors for a prolonged period f 8 days), leading to more potent activation of adjacent substance P receptor expressing neurons. Furthermore, repealed stimulation leads to greater substance P release, resulting in greater diffusion of substance P away from the perivascular site of release and the stimulation of more distant neurons. It has been estimated that approximately 3-5 times more neurons are activated in response to more frequent stimulation (Mantyh, 2002, J, Clin. Psychiatry 63. 6-10). As shown by us in example 3, administration of a substance P receptor antagonist prevents the acti vation of the substance P receptor, prevents the activation of enzymes responsible for tau hyperphosphorylation, and thus prevents such tau hyperphosphorylation and the development/progression of chronic traumatic encephalopathy.

]¾ampje 5

Concussion results in the. greatest mechanical stress at (he base of sulci

Having established that both release of substance P and accumulation of hyperphosphorylated ta occurred following a concussive event, it remained to be shown why accumulation of hyperphosphorylated tau in human CTE was prominent al the base of the brain, sulci. We have shown in example two that substance P release occurs in response to stimulation of meehanoreceptors on sensory nerve fibres, One highly innovative step in the current disclosure is the realization that in gyrencephalic animals, including man, sulci protect the brain cortex from mechanical injury by focusing the stress points to the base of the sulci. This is best illustrated in models of brain tissue thai both incorporate and exclude sulci within them, and subsequently simulating the effects of mechanical stress (Cioots et a!., 2008, Ann. Biomed. Eng, 36, 1203-1215). The simulation of the effects of mechanical strain typical of rotational acceleration in models replicating- the brain tissue with and without sulci is shown in Figure 5, Higher mechanical stress is indicated as black. These results clearly show that the addition of sulci to the model focuses the stress to the base of the sulci, irrespective of the sulcus morphology. This mechanical stress pattern is remarkably similar to the post-mortem localisation: of hyperphosphorylated tau reported in CTE and shown in Figure 1 ,

This example confirms that in a gyreneephalie brain, exposure to a mechanical eoneussive event will focus the mechanical forces to the base of the. sulci, thereby preferentially activating mec anoreceptorS. on sensory nerves in that location. Example 2 has already shown that activation of mechanoreceptors on sensory nerve fibres will cause the perivascular release of substance P.