Neurodegenerative Therapies

  *US20170007583A1*
  US20170007583A1                                 
(19)United States 
(12)Patent Application Publication(10)Pub. No.: US 2017/0007583 A1
 (43)Pub. Date:Jan.  12, 2017

(54)Neurodegenerative Therapies 
    
(76)Inventor: LYTIX BIOPHARMA AS,  Tromsø (NO) 
(21)Appl. No.: 15/116,296 
(22)PCT Filed:Feb.  4, 2015 
(86)PCT No.: PCT/EP2015/052330 
(30)Foreign Application Priority Data 
 Feb.  4, 2014(GB)1401886.5
 Publication Classification 
(51)Int. Cl. A61K 031/428 (20060101); C12N 009/99 (20060101); A61K 031/4439 (20060101)
CPC A61K 031/428 (20130101); A61K 031/4439 (20130101); C12N 009/99 (20130101)

        

(57)

Abstract

The present invention provides a compound of formula (I) wherein: Y represents a C or N atom which may be substituted or form a cyclic group with R′″ but may not be a quaternary C atom; R′ is —OR1, —CONH2, —CF3, F, —OH, —NO2, —CN or —OCOR1 in which R1, is C1-3 alkyl and each may be in the beta or gamma position; R″ is C1-3 alkyl or H; and R′″ is H or a group consisting of 1-12 non-hydrogen atoms and may be linear, branched and/or incorporate one or more cyclic groups, cyclic groups may be aromatic and/or heterocyclic and 2 or more cyclic groups may be linked or fused and each may be substituted; or a salt, hydrate or solvate of a compound of formula (I) for use in the treatment or prevention of a neurodegenerative disorder by inhibiting formation of neurofibrillary (tau) tangles and/or by inhibiting Dyrk 1A. The invention further relates to non-therapeutic uses of these compounds.
[see pdf for image]
 Claim(s),  Drawing Sheet(s), and Figure(s)
 
 
[0001] The present invention relates to the field of neurodegenerative disorders, in particular to Alzheimer's disease (AD).
[0002] AD is the most common form of dementia and no treatment exists which can stop, let alone reverse, progression of the disease. The memory and other mental health implications of AD are well known but the disease is also a killer; the average life expectancy after diagnosis is about 7 years as bodily functions are gradually lost. This is a common degenerative condition, generally affecting people over 65, and it is recognised as placing a significant burden on careers, health services and society in general as life expectancy continues to rise and the numbers of people affected by AD increases.
[0003] AD is characterised by loss of neurons and synapses in the cerebral cortex and some subcortical regions. Amyloid plaques and neurofibrillary tangles are observed in the brains of those with AD.
[0004] Amyloid plaques form on the outside of neurones and are made up of peptides of 39-43 amino acids called beta-amyloid (Aβ), these are fragments of amyloid precursor protein, a trans-membrane protein that penetrates the neuron's membrane and is critical to neuron growth, survival and repair.
[0005] Neurofibrillary tangles are aggregates of the microtubule-associated protein tau which have become hyperphosphorylated and accumulated in the neurons. In healthy neurones, tau serves to stabilise the microtubules of the neuronal cytoskeleton. Certain conditions are characterised by an increase in these tau tangles and this group of conditions are referred to as tauopathies. Tauopathies include Parkinson's disease, Pick's disease and progressive supernuclear palsy, as well as AD. While an increase in amyloid plaques may be seen decades before the onset of symptoms of AD, the symptoms of AD are often observed just after a noticeable increase in tau protein is seen.
[0006] While it is generally accepted that these two proteins have a role in AD, the pathological mechanism and the causal events are not known. It had been postulated that the formation of amyloid plaques caused AD but therapies which successfully reduced plaque formation did not give significant improvement in symptoms such as dementia.
[0007] Current medication for AD shows limited benefit. Acetycholinesterase inhibitors such as tacrine and donepezil are used to decrease the rate at which acetylcholine (ACh) is broken down in the brain, in order to counteract the reduction in cholinergic neuron activity which is associated with AD. These therapies have shown some benefit, at least in mild to moderate AD. The NMDA receptor antagonist memantine has been shown to have very modest efficacy in the treatment of moderate to severe AD.
[0008] There is undoubtedly an urgent need for further therapeutic options in the treatment of AD and other neurodegenerative conditions, whether to slow or halt disease progression, improve symptoms or delay onset; the tools available to the clinician at present are completely inadequate.
[0009] The present inventors have developed compounds which act as inhibitors of Dyrk1A (dual-specificity tyrosine-(Y)-phosphorylation-regulated kinase 1A), a kinase thought to be important in neonates and in the early stages of life. Dyrk1A is a kinase whose over-activity has recently been implicated in the pathogenesis of AD and other tauopathies. The Dyrk1A gene is copied in triplicate in patients that have Down Syndrome (DS), who are themselves more likely to develop AD; between 50 and 70% of DS patients develop dementia by age 60 and nearly all DS patients have amyloid plaques and neurofibrillary tangles above the age of 30. Dyrk1A is thought to play a role in the development of AD, both by increasing amyloid plaque formation and increasing intracellular tau protein tangles. Studies have identified Dyrk1A as the priming kinase of multiple phosphorylation of the tau protein and studies of the brains of patients with AD showed increased expression of Dyrk1A in neurons affected by tau tangles.
[0010] Ogawa et al. in Nature Communications, 5 Oct. 2010 (1) Article 86 describe a Dyrk1A inhibitor called INDY, a benzothiazoylidene. INDY binds at the ATP binding cleft. INDY is not well able to cross cell membranes and has the complication that it must be administered as a prodrug. Harmine is a potent inhibitor of Dyrk1A but is hallucinogenic.
[0011] Thus there is a need for alternative and/or improved inhibitors of Dyrk1A. In particular for highly selective kinase inhibitors so that off-target effects on other kinases are reduced. It is also necessary for the compounds to pass the blood brain barrier. The present inventors have identified a new class of Dyrk1A inhibitor which possess some or all of these advantageous features.
[0012] According to one aspect, the present invention provides a compound of formula (I)
[see pdf for image]
wherein:
Y represents a C or N atom which may be substituted or form a cyclic group with R′″ but may not be a quaternary C atom;
R′ is —OR1, —CONH2, —CF3, F, —OH, —NO2, —CN or —OCOR1 in which R1 is C1-3 alkyl and each may be in the beta or gamma position;
R″ is C1-3 alkyl or H; and
R′″ is H or a group consisting of 1-12 non-hydrogen atoms and may be linear, branched and/or incorporate one or more cyclic groups, cyclic groups may be aromatic and/or heterocyclic and 2 or more cyclic groups may be linked or fused and each may be substituted;
or a salt, hydrate or solvate of a compound of formula (I)
for use in the treatment or prevention of a neurodegenerative disorder by inhibiting formation of neurofibrillary (tau) tangles.
[0013] The compounds of formula (I) are Dyrk1A inhibitors and preferably can cross the blood brain barrier. In a further aspect the present invention provides a compound of formula (I) as defined herein, or a salt, hydrate or solvate of a compound of formula (I), for use in the treatment or prevention of a neurodegenerative disorder by inhibiting Dyrk1A.
[0014] A quaternary C atom is one bonded to 4 other C atoms.
[0015] In a further aspect, the present invention provides a method of treating or preventing a neurodegenerative disease in a subject comprising administering a therapeutically effective amount of a compound of formula (I)
[see pdf for image]
wherein:
Y represents a C or N atom which may be substituted or form a cyclic group with R′″ but may not be a quaternary C atom;
R′ is —OR1, —CONH2, —CF3, F, —OH, —NO2, —CN or —OCOR1 in which R1 is C1-3 alkyl and each may be in the beta or gamma position;
R″ is C1-3 alkyl or H; and
R′″ is H or a group consisting of 1-12 non-hydrogen atoms and may be linear, branched and/or incorporate one or more cyclic groups, cyclic groups may be aromatic and/or heterocyclic and 2 or more cyclic groups may be linked or fused and each may be substituted;
or a salt, hydrate or solvate of a compound of formula (I)
to said subject, with the proviso that the compound of formula (I) is not:
[0016] (i) one of the following compounds of formula (II)
[see pdf for image]
[0017] in which R is benzyl substituted as follows:
[00001] [TABLE-US-00001]
 
  Compound   Benzyl substituent
 
  (a)   4-OH-5-COOH
  (b)   2-OH
  (c)4-OH-5-COOCH3
  (d)2-I-4-COOCH3
 
[0018] or (e) R is pyridyl, or
[0019] (ii) selected from
[0020] (f) N-(6-cyano-2-benzothiazolyl)-N-[(4-methoxyphenyl)methyl]-urea
[0021] (g) ethyl 2-(3-(6-fluorobenzo[d]thiazole-2-yl)ureido)acetate
[0022] (h) ethyl 2-(3-(6-methoxybenzo[d]thiazole-2-yl)ureido)acetate
[0023] (i) potassium 2-(3-(6-methoxybenzo[d]thiazole-2-yl)ureido)acetate
[0024] (j) 2-(3-(6-methoxybenzo[d]thiazole-2-yl)ureido)acetic acid
[0025] (k) N-(2,6-dimethylphenyl)-N-(6-fluoro-2-benzothiazolyl)urea
[0026] (l) benzeneacetamide, 3,5-difluoro-N-[(1S)-1-[[(6-methoxy-2-benzothiazolyl)amino]carbonyl]propyl].
[0027] A therapeutically effective amount is preferably one which is able to inhibit formation of neurofibrillary (tau) tangles (and thereby treat or prevent a neurodegenerative disorder). A therapeutically effective amount is preferably one which inhibits Dyrk1A (and thereby treats or prevents a neurodegenerative disorder).
[0028] Preferably the compounds of formula (I) for use according to, and in the methods of, the present invention do not include the following compounds:
[0029] (m) 2-(3,5-dimethylphenoxy)-N-(6-nitrobenzothiazol-2-yl) acetamide
[0030] (n) 1-(6-fluorobenzothiazol-2-yl)-3-(4-fluorophenyl)urea
[0031] (o) 1-(4-fluorophenyl)-3-(6-methoxybenzothiazol-2-yl) urea
[0032] (p) 4-[3-(6-fluorobenzothiazol-2-yl)ureido]benzoic acid ethyl ester
[0033] (q) benzeneacetamide, α-ethyl-N-(6-methoxy-2-benzothiazolyl)
[0034] Y is preferably carbon and preferably forms a cyclic group with R′″, preferably an aromatic cyclic group. If Y forms a cyclic group with R′″ then together the moiety —YR′″ consists of no more than 13 non-hydrogen atoms. If Y is substituted, it is preferably by a C1-3 alkyl group but in other preferred embodiments it is unsubstituted. If the Y atom is unsubstituted and not part of a cyclic group with R′″, then Y is —CH2— or —NH—.
[0035] R″ is preferably H or methyl, most preferably H.
[0036] R′ is preferably attached to the 6 carbon atom unless it is —CF3, —CONH2, —NO2 or —CN, in which case it is preferably attached to the γ carbon atom. —CF3, —NO2 and —CN are particularly preferred in the γ position. Preferably R′ is not F.
[0037] R1 is preferably methyl and thus —OCH3 is a preferred R′ moiety, if R′ is —OCH3 it is preferably attached to the β carbon atom.
[0038] R′″ may, in certain preferred embodiments, be hydrogen and in this case YR′″ together are preferably —NH2 or —CH3, most preferably CH3.
[0039] In a further preferred embodiment R′″ is C1-5 alkyl, which may be linear or branched, or a derivative thereof which may optionally be substituted, e.g. by oxygen, nitrogen or a halogen. Preferred alkyl derivatives are those in which one of the carbon atoms is replaced by a nitrogen or oxygen atom. Thus, for example, R′″ may be —CONH2, —CH2COCH3, —CH2CHNH2CH3, —NHCOCH3, —CH2CH3, —CH2CH2CH3, —COCH3, —COCH2CH3, —CONHCH3.
[0040] In further preferred embodiments R′″ is an amino acylated amino acid or an acylated dipeptide. The acyl group is preferably acetyl. The amino acid(s) are preferably genetically encoded amino acids, more preferably selected from alanine, phenylalanine, tyrosine, proline or serine.
[0041] Alternatively, R′″ or YR′″ together may contain 1-3, preferably 1 or 2 cyclic groups, most preferably 1 cyclic group. For example, R′″ or YR′″ together may consist of one cyclic group of 3 to 7 ring atoms, preferably 3, 5 or 6 ring atoms, where at most 3 ring atoms are non-carbon atoms. Preferred non-carbon atoms are nitrogen, oxygen and sulphur, more preferably nitrogen and oxygen, most preferably nitrogen. The cyclic group may or may not be aromatic, but an aromatic ring is preferred.
[0042] The cyclic group is optionally substituted with one or more substituents containing a total of up to 8 non-hydrogen atoms. The substituents may be small (e.g. C1-3) alkyl groups including cyclopropyl, but are preferably polar in nature. Examples of suitable substituents are presented in Table A below.
[0043] In alternative embodiments, R′ or YR′″ together may contain two fused cyclic groups, for example two fused five membered rings, one five membered ring fused to a six membered ring or two fused six membered rings. The fused rings together may contain 0-5 non-carbon atoms (e.g. 1-3 non-carbon atoms), preferably nitrogen, oxygen or sulphur, more preferably nitrogen or oxygen, most preferably nitrogen. Neither, either or both, preferably both, of the fused rings may be aromatic.
[0044] One or more of the fused cyclic groups are optionally substituted with one or more substituents containing a total of up to 5 non-hydrogen atoms. The substituents may be small (e.g. C1-3) alkyl groups, including cyclopropyl, but are preferably polar in nature. Examples of suitable substituents are presented in Table A below.
[0045] In alternative embodiments, R″ or YR′″ together may contain two linked cyclic groups. The link may be direct forming a biaryl system or through a linking atom, usually carbon, nitrogen or oxygen. The linked rings together may contain 0-5 non-carbon atoms (e.g. 1-3 non-carbon atoms), preferably nitrogen, oxygen or sulphur, more preferably nitrogen or oxygen, most preferably nitrogen. Neither, either or both, preferably both, of the linked cyclic groups may be aromatic.
[0046] One or more of the linked cyclic groups are optionally substituted with one or more substituents containing up to 3 non-hydrogen atoms. The substituents may be small alkyl groups (e.g. C1-3), including cyclopropyl, but are preferably polar in nature. Examples of suitable substituents are presented in Table A below.
[00002] [TABLE-US-00002]
  TABLE A
 
  Preferred substituents to the cyclic group(s) of —R′″ or
  —YR′″, ordered by the number of non-hydrogen atoms:
  No. of  
  heavy atoms   Examples of suitable substituents
 
  1   Methyl, halogen (F, Cl, Br), hydroxyl, amino and
    sulfhydryl
  2   Ethyl, ethenyl (vinyl), ethynyl, methoxy, methylamino,
    methylsulfide, cyano and formyl
  3   Propyl, isopropyl, cyclopropyl, methylsulfoxy, acetyl,
  nitro, dimethylamino, CH2CH2OH, CH2CH2NH2,
    carboxylate, carboxamide
  4—OCH2CH2OH, —NHCH2CH2OH, —OCH2CH2NH2,
  —NHCH2CH2NH2, methylcarboxylate, N-
    methylcarboxamide, trifluoromethyl, methylsulfonyl,
    sulphonamide sulfonic acid
  5   Methylsulfonylamido, trifluoromethoxy,
    cyclopropylmethoxy, N,N-dimethylcarboxamido
  6piperidyl, morpholinyl, —C═ONHCH2CH2NH2,
  —C═ONHCH2CH2OH, C═OOCH2CH2OH
  7N(CH2CH2NH2)2, N(CH2CH2OH)2, methylpiperidyl
 
[0047] Preferred linking groups include —CH2—, —NR—, —O— and —S—, wherein R is H or C1-2 alkyl.
[0048] If R′″ or YR′″ together contain two or more cyclic groups these may be the same or different and if they are substituted, the substituents may be the same or different.
[0049] If R′″ or YR′″ together comprise two or more aromatic groups, then preferably no more than one of these groups is carbocyclic.
[0050] Suitable cyclic groups for R′″ or YR′″ together include cyclohexyl, phenyl, pyridine, pyrimidine, pyrazole, imidazole, thiazole, oxazole, diazolone, morpholine and piperidine.
[0051] Preferred substituents of the cyclic group(s) of R′″ (or YR′″) include: —OH, —OR, —NRR, —C═O—OR, —C═O—NRR, —SO2—NRR, —NR—SO2R, —SO2R, —NR—C═OR and a halogen (e.g. F or Cl), wherein each R (which may be the same or different) is H or C1-3 alkyl, preferred substituents are C1-3 alkyl or a derivative thereof which may itself be substituted (e.g. by oxygen, —CH3, NH2 or a halogen such as F).
[0052] In preferred embodiments, YR′″ together form a single cyclic group, preferably aromatic and preferably 6 membered, most preferably, pyrimidinyl, pyridyl or phenyl; the cyclic group is optionally and preferably substituted, e.g. by NH2 or a C1-3 alkyl or derivative thereof which may itself be substituted e.g. by oxygen, nitrogen or a halogen. Preferred alkyl derivatives are those in which one of the carbon atoms is replaced by a nitrogen or oxygen atom. The substituent of the cyclic group may preferably be selected from the group comprising —CONH2, —CH2COCH3, —CH2CHNH2CH3, —NHCOCH3, —CH2CH3, —CH2CH2CH3, —OCH3, —COCH3, —COCH2CH3 and —CONHCH3.
[0053] Pharmaceutically acceptable salts, hydrates and solvates are well known in the art.
[0054] Neurodegenerative disorders according to the present invention include Alzheimer's disease, Parkinson's disease, Pick's disease, progressive supranuclear palsy, corticobasal degeneration and agyrophilic grain disease. In particular the present invention concerns the treatment or prevention of Alzheimer's disease, more particularly the treatment or prevention of Alzheimer's disease in patients with Down Syndrome. Tauopathies are a particular target for treatment according to the present invention.
[0055] Treatment includes an improvement in one or more of the symptoms of the disorder or a delay in onset of one or more symptoms as assessed by a clinician, optionally together with patient feedback. Symptoms of AD include memory loss, confusion, mood and personality changes, hallucinations, delusions and paranoia, problems with communication, weight loss, seizures, skin infections, difficulty in swallowing and lack of control of bowel and bladder.
[0056] Treatment includes slowing or halting disease progression and thus treatment may not result in significant observable benefits unless a comparison is made with expected (untreated) progression of the disorder. Likewise, treatment may be beneficial if an anticipated symptom is delayed in its appearance.
[0057] The subject will typically have been identified as in need of treatment. This may be determined based on assessment of cognitive performance or any other measure which leads to a diagnosis that the patient has a neurodegenerative disorder or is at risk of developing such a disorder. In the case of AD this determination may be achieved through microscopic histological or other investigations to observe the formation of amyloid plaques and/or neurofibrillary tangles.
[0058] Prevention of a neurodegenerative disorder may include prevention for a period of time, in other words delayed onset. Suitable patients for prevention include those with DS, in particular, DS patients over the age of 20 or 30. Generally, if a patient has been shown to have one or more markers of a neurodegenerative disorder but no symptoms as yet, such a patient is considered to be “treated” in accordance with the present invention. “Prevention” assumes the patient has neither symptoms nor confirmed clinical markers of disease.
[0059] The present invention typically involves inhibition of the formation of neurofibrillary tangles. Without wishing to be bound by theory, this is believed to be key to the clinical success of the present invention and at least part of the mechanism by which the disclosed Dyrk1A inhibitors are effective against neurodegenerative disorders. These (tau) tangles can be assessed by any convenient method known in the art, for example using a microscope to observe the aggregates of the tau protein, a suitable method is described by Armstrong in Folia Neuropathol. 2008; 46 (1): 26-31.
[0060] Inhibition may be observed on treatment through a reduction in the size of the tangles or in the extent of their distribution. Their formation is “inhibited” even if the amount observed has not decreased on treatment, if the amount would have been expected to increase without treatment. Of course, such examinations of a patient may not be possible during a treatment regimen, but suitable studies can easily be performed to select a compound from within the definition of compounds of formula (I).
[0061] Alternatively viewed, the compounds described herein treat or prevent neurodegenerative disorders through inhibition of Dyrk1A. A method to measure inhibition of Dyrk1A is described in the Examples hereto. A suitable assay could be performed using ADP-Glo™ kinase assay of Promega. Compounds of the invention preferably can achieve at least 30, more preferably at least 50 or 60, most preferably at least 70 or 80% inhibition of Dyrk1A in such assays.
[0062] In a further aspect, the present invention provides a method of inhibiting Dyrk1A and/or a reaction catalysed by Dyrk1A, the method comprising contacting said kinase with a compound of formula (I) as defined herein or a salt, hydrate or solvate of a compound of formula (I). Such methods may be in vivo or ex vivo, e.g. in vitro.
[0063] In a further aspect, the present invention provides the use of a compound of formula (I) or a salt, hydrate or solvate of a compound of formula (I) in the manufacture of a medicament for the treatment of a neurodegenerative disorder.
[0064] Animals which may be treated include domestic animals, in particular cats and dogs and livestock animals such as pigs, cows, sheep or goats as well as horses. Laboratory animals, mice, rabbits etc. may also be treated. Treatment of humans is nevertheless preferred.
[0065] Methods for the synthesis of compounds of the invention are described in the Examples hereto, non-exemplified compounds can be prepared by methods which are analogous to the schemes and protocols described herein.
[0066] Methods of synthesising compounds of the invention, in particular methods described in the Examples, constitute a further aspect of the present invention.
[0067] All novel compounds, defined generically by the formulae herein or individual compounds recited, in particular in the Examples, constitute a further aspect of the present invention.
[0068] The compositions according to the invention may be presented, for example, in a form suitable for oral, nasal, parenteral, intravenal, topical or rectal administration.
[0069] The active compounds defined herein may be presented in the conventional pharmacological forms of administration, such as tablets, coated tablets, nasal sprays, inhalers, solutions, emulsions, liposomes, powders, capsules or sustained release forms. As used herein, the term “pharmaceutical” includes veterinary applications of the products of the invention.
[0070] Conventional pharmaceutical excipients as well as the usual methods of production may be employed for the preparation of these forms. Tablets may be produced, for example, by mixing the active ingredient or ingredients with known excipients, such as for example with diluents, such as calcium carbonate, calcium phosphate or lactose, disintegrants such as corn starch or alginic acid, binders such as starch or gelatin, lubricants such as magnesium stearate or talcum, and/or agents for obtaining sustained release, such as carboxypolymethylene, carboxymethyl cellulose, cellulose acetate phthalate, or polyvinylacetate.
[0071] The tablets may if desired consist of several layers. Coated tablets may be produced by coating cores, obtained in a similar manner to the tablets, with agents commonly used for tablet coatings, for example, polyvinyl pyrrolidone or shellac, gum arabic, talcum, titanium dioxide or sugar. In order to obtain sustained release or to avoid incompatibilities, the core may consist of several layers too. The tablet coat may also consist of several layers in order to obtain sustained release, in which case the excipients mentioned above for tablets may be used.
[0072] Injection solutions may, for example, be produced in the conventional manner, such as by the addition of preservation agents, such as p-hydroxybenzoates, or stabilizers, such as EDTA. The solutions are then filled into injection vials or ampoules.
[0073] Nasal sprays administration may be formulated similarly in aqueous solution and packed into spray containers either with an aerosol propellant or provided with means for manual compression.
[0074] Capsules containing one or several active ingredients may be produced, for example, by mixing the active ingredients with inert carriers, such as lactose or sorbitol, and filling the mixture into gelatin capsules.
[0075] Suitable suppositories may, for example, be produced by mixing the active ingredient or active ingredient combinations with the conventional carriers envisaged for this purpose, such as natural fats or polyethyleneglycol or derivatives thereof.
[0076] Tablets for oral administration are preferred.
[0077] Pharmaceutical compositions comprising a compound of formula (I) may additionally comprise further active ingredients, including, for example, other active agents for the treatment or prevention of a neurogenerative disorder. Likewise the medical uses and methods of treatment may additionally comprise further active ingredients, including, for example, other active agents for the treatment or prevention of a neurogenerative disorder, e.g AD.
[0078] In employing such compositions systemically (intra-muscular, intravenous, intraperitoneal), the active molecule is generally present in an amount to achieve a serum level of the active molecule of at least about 1-10 micromolar Such serum levels may be achieved by incorporating the bioactive molecule in a composition to be administered systemically at a dose of from 50 mg-250 mg.
[0079] It is appreciated that appropriate dosages will vary from patient to patient dependent on age, sex, previous treatments, severity of symptoms presented etc.
[0080] The above description describes numerous features of the present invention and in most cases preferred embodiments of each feature are described. It will be appreciated that each preferred embodiment of a given feature may provide a molecule, use, method etc. of the invention which is preferred, both when combined with the other features of the invention in their most general form and when combined with preferred embodiments of other features. The effect of selecting multiple preferred embodiments may be additive or synergistic. Thus all such combinations are contemplated unless the technical context obviously makes them mutually exclusive or contradictory. In general each feature and preferred embodiments of it are independent of the other features and hence combinations of preferred embodiments may be presented to describe sub-sets of the most general definitions without providing the skilled reader with any new concepts or information as such.
[0081] The invention will now be further described with reference to the following non-limiting Examples. Not all of the compounds synthesised and tested are within the scope of the present invention, some are provided for comparative purposes and highlight the efficacy of the molecules of the invention.

EXAMPLES

A. Preparation of the Compounds of the Invention

Overview

[0082] The amide compounds of the invention can conveniently be prepared by acylation of 2-aminobenzo-1,3-thiazoles. The acylation could be performed by treating the 2-aminobenzo-1,3-thiazoles with acetyl chloride or acetyl anhydride (R═CH3) or by treatment of a carboxylic acid (R≠CH3) under coupling conditions (e.g. using HBTU or other coupling reagents typically used for peptide synthesis).
[see pdf for image]
[0083] The 2-aminobenzo-1,3-thiazoles themselves can be prepared in a two-step sequence from the corresponding aniline as outlined below. The aniline derivative is treated with potassium thiocyanate forming a thiourea derivative that is subsequently cyclised by addition of bromine.
[see pdf for image]
[0084] Compounds with a “reversed” amide bond, included as comparative examples, can be prepared from 2-benzo-1,3-thiazole carboxylic acid derivatives by performing a HBTU mediated coupling to a suitable amine or aniline derivative.
[see pdf for image]
[0085] Urea derivatives can be prepared by the reaction between 2-aminobenzo-1,3-thiazole derivatives and suitable alkyl- or aryl isocyanates as shown in the scheme below.
[see pdf for image]

Example 1

N-(5-hydroxybenzo[d]thiazol-2-yl)acetamide

[0086]  [see pdf for image]

Step 1. 1-(3-methoxyphenyl)thiourea

[0087] 3-Methoxyaniline (approx 57 mmol) was added to a mixture of sodium thiocyanate (15 eqv) in isopropylacetate (56 mL) at room temperature. Trifluoroacetic acid (11 mL (2.5 eqv)) was then added portionwise ensuring that the temperature of the mixture was maintained at 40° C. or less. The temperature of the mixture was then raised to 85° C. and stirred for 16 hours. The mixture was cooled and 7 mL of distilled water were added before further cooling to 0° C. The crude product was isolated by vacuum filtration. The crude yield was in the range of 80%

Step 2. 5-Methoxybenzo(dithiazol-2-amine

[0088] The thiourea above (5.5 mmol) was dissolved in acetic acid (10 mL) and lithium bromide (1.5 eqv) was added at room temperature. Bromine (1 eqv) was added portionwise (the reaction is very exothermic) ensuring that the temperature mixture did not exceed 30° C. The reaction vessel was then heated to 40° C. The reaction is stirred and subsequently cooled room temperature. The product is isolated by vacuum filtration and washed twice with 5% sodium carbonate solution and twice with distilled water before drying. The crude yield was in the range of 75%.

Step 3. 5-Hydroxyoxybenzo[d]thiazol-2-amine

[0089] Borontribromide (2 ml) was added to a suspension of 5-methoxybenzo[d]thiazol-2-amine (0.18 mmol) in dichlorormethane at 0° C. before overnight stirring at ambient temperature. The reaction mixture was carefully quenched with water before extraction with ethyl acetate. The crude product was isolated by evaporation of the solvent under reduced pressure.

Step 4. N-(5-Hydroxybenzo[d]thiazol-2-yl)acetamide

[0090] The 2-aminobenzothiazole above (0.5 mmol) and acetic anhydride (2.2 eqv.) were dissolved in 2 mL of DMF. DIPEA (3 eqv) was added and the mixture was placed in a microwave oven and irradiated at 60° C. for 2 h. The mixture was triturated into 50 mL of 5% NaHSO4 and stirred for 15 min. The crude product was hydrolysed in 5% NaOH before pH adjustment to 7-7.5 where the N-acetylated product precipitated.
[0091] In a similar manner the following products were prepared:
[00003] [TABLE-US-00003]
 
[see pdf for image]
 
      Calcd mass   Obsd mass   1H NMR
  X   Compound   <MH+>   <MH+>   (Exchangeable protons not reported)
 
  γ-OH   01-71   209.03   208.91H NMR (400 MHz, Methanol-d4) δ 7.54 (d, J =
          8.7 Hz, 1H), 7.20 (d, J = 2.4 Hz, 1H), 6.90 (dd, J =
          8.8, 2.5 Hz, 1H), 2.23 (s, 3H).
  β-OH   01-66   209.03   208.91H NMR (400 MHz, Methanol-d4) δ 7.62 (d, J =
          8.6 Hz, 1H), 7.14 (d, J = 2.3 Hz, 1H), 6.83 (dd, J =
          8.6, 2.3 Hz, 1H), 2.25 (s, 3H).
γ-CF3   01-90   261.03   260.81H NMR (400 MHz, DMSO-d6) δ 8.45 (d, J = 1.7
          Hz, 1H), 7.88 (d, J = 8.5 Hz, 1H), 7.71 (dd, J = 8.5,
          1.9 Hz, 1H), 2.21 (s, 3H).
β-CF3   03-73   261.03   260.81H NMR (400 MHz, Methanol-d4) δ 8.04 (d, J =
          8.3 Hz, 1H), 8.01-7.95 (m, 1H), 7.59-7.49 (m, 1H),
          2.27 (s, 3H).
  γ-CN   01-94   218.03   217.91H NMR (400 MHz, DMSO-d6) δ 8.53 (d, J = 1.6
          Hz, 1H), 7.91-7.64 (m, 2H), 2.22 (s, 3H).
β-CH3    03-76*   207.05   207.21H NMR (400 MHz, DMSO-d6) δ 7.80 (s, 1H),
          7.53 (s, 1H), 7.16-7.07 (m, 1H), 2.39 (s, 3H),
          2.17 (s, 3H).
  β-F   03-77   211.03   211.31H NMR (400 MHz, DMSO-d6) δ 7.96 (dd,
          J = 8.7, 5.5 Hz, 1H), 7.53 (dd, J = 10.1, 2.4 Hz,
          1H), 7.16 (td, J = 9.0, 2.5 Hz, 1H), 2.19 (s, 3H).
  H    03-32*   193.04   193.01H NMR (400 MHz, DMSO-d6) δ 7.94
          (s, 1H), 7.71 (s, 1H), 7.41 (s, 1H), 7.28 (s, 1H),
          2.18 (s, 3H).
β-OCH3   03-21   223.05   223.01H NMR (400 MHz, DMSO-d6) δ 7.79 (d, J = 8.7
          Hz, 1H), 7.25 (d, J = 2.4 Hz, 1H), 6.91 (dd, J = 8.8,
          2.4 Hz, 1H), 3.80 (s, 4H), 2.17 (s, 3H).
  γ-F   03-94   211.03   210.41H NMR (600 MHz, DMSO-d6) δ 12.36 (s, 1H),
          7.87 (dd, J = 8.8, 2.8 Hz, 1H), 7.73 (dd, J = 8.9, 4.8
          Hz, 1H), 7.27 (td, J = 9.1, 2.8 Hz, 1H), 2.20 (s, 3H).
γ-NO2   13-10   238.02   237.61H NMR (400 MHz, DMSO-d6) δ 9.04 (d, J = 2.5
          Hz, 1H), 8.27 (dd, J = 9.0, 2.4 Hz, 1H), 7.88 (d, J =
          8.9 Hz, 1H), 2.25 (s, 3H).
γ-OCH3   03-20   223.05   223.01H NMR (600 MHz, DMOS-d6) δ 12.19 (s, 1H),
          7.62 (d, J = 8.8 Hz, 1H), 7.54 (d, J = 2.6 Hz, 1H),
          7.01 (dd, J = 8.8, 2.6 Hz, 1H), 3.80 (s, 3H), 2.18
          (s, 3H).
γ-CONH2   08-70   236.05   235.61H NMR (600 MHz, DMSO-d6) δ 8.44 (d, J = 1.8
          Hz, 1H), 7.92 (dd, J = 8.5, 1.8 Hz, 1H), 7.74 (d, J =
          8.4 Hz, 1H), 2.20 (s, 3H).
 
  * = comparative example

Example 2

Methyl 3-((6-(trifluoromethyl)benzo[d]thiazol-2-yl)carbamoyl)benzoate

[0092]  [see pdf for image]
[0093] 6-(trifluoromethyl)benzo[d]thiazol-2-amine (commercially available or prepared according to Step 1 and 2 of Example 1) (1 eq), 3-(methoxycarbonyl)benzoic acid (1.05 eq) and DIPEA (5 eq) were dissolved in DMF (2 ml). HBTU (1.2 eq) was added and the reaction were stirred overnight at ambient temperature before extraction with ethylacetate, washed thoroughly 2 times with dilute acid and dried. The crude product, methyl 3-((6-(trifluoromethyl)benzo[d]thiazol-2-yl)carbamoyl)benzoate, was isolated by evaporation of the solvent under reduced pressure.
[0094] In a similar manner the following products were prepared:
[00004] [TABLE-US-00004]
 
[see pdf for image]
 
        Calcd mass   Obsd mass   1H NMR
  X   R   Compound   <MH+>   <MH+>   (Exchangeable protons not reported)
 
  γ-OH [see pdf for image]   08-42   272.04   272.01H NMR (400 MHz, DMSO-d6) δ 8.84 (d, J = 5.4 Hz, 2H), 8.05 (d, J = 5.5 Hz, 2H), 7.58 (d, J = 8.7 Hz, 1H), 7.32 (d, J = 2.4 Hz, 1H), 6.92 (dd, J = 8.7, 2.4 Hz, 1H).
 
  γ-OH [see pdf for image]   08-44   272.04   272.11H NMR (400 MHz, DMSO-d6) δ 9.24 (d, J = 2.2 Hz, 1H), 8.82 (dd, J = 5.0, 1.6 Hz, 1H), 8.52 (d, J = 8.1 Hz, 1H), 7.66 (dd, J = 8.0, 4.9 Hz, 1H), 7.57 (d, J = 8.7 Hz, 1H), 7.31 (d, J = 2.4 Hz, 1H), 6.91 (dd, J = 8.7, 2.4 Hz, 1H).
 
  γ-OH [see pdf for image]   03-12   272.04   272.11H NMR (400 MHz, Methanol-d4) δ 8.78 (d, J = 4.7 Hz, 1H), 8.33 (d, J = 7.9 Hz, 1H), 8.13 (d, J = 8.3 Hz, 1H), 7.74 (d, J = 7.0 Hz, 1H), 7.61 (d, J = 8.8 Hz, 1H), 7.27 (d, J = 2.3 Hz, 1H), 6.96 (dd, J = 8.8, 2.5 Hz, 1H).
 
  β-OH [see pdf for image]   03-11   272.04   271.71H NMR (400 MHz, DMSO-d6) δ 8.86 (d, J = 5.5 Hz, 2H), 8.10 (d, J = 5.2 Hz, 2H), 7.75 (d, J = 8.6 Hz, 1H), 7.10 (d, J = 2.3 Hz, 1H), 6.84 (dd, J = 8.6, 2.2 Hz, 1H).
 
  β-OH [see pdf for image]   03-33   272.04   272.11H NMR (400 MHz, DMSO-d6) δ 9.23 (d, J = 2.3 Hz, 1H), 8.76 (d, J = 4.8 Hz, 1H), 8.52-8.28 (m, 1H), 7.70 (d, J = 8.5 Hz, 1H), 7.56 (dd, J = 8.0, 4.8 Hz, 1H), 7.08 (d, J = 2.4 Hz, 1H), 6.79 (dd, J = 8.4, 2.4 Hz, 1H).
 
  β-OH [see pdf for image]   03-13   272.04   272.11H NMR (400 MHz, Methanol-d4) δ 8.82 (d, J = 5.2 Hz, 1H), 8.50 (d, J = 8.0 Hz, 1H), 8.36 (d, J = 8.0 Hz, 1H), 7.90 (t, J = 6.6 Hz, 1H), 7.69 (d, J = 8.7 Hz, 1H), 7.14 (d, J = 2.3 Hz, 1H), 6.92 (dd, J = 8.7, 2.3 Hz, 1H).
 
γ-CF3 [see pdf for image]   03-99   323.04   322.71H NMR (400 MHz, Methanol-d4) δ 8.55 (s, 1H), 8.16 (d, J = 7.7 Hz, 2H), 7.95 (d, J = 8.4 Hz, 1H), 7.78 (d, J = 8.5 Hz, 1H), 7.69 (t, J = 7.3 Hz, 1H), 7.58 (t, J = 7.7 Hz, 2H).
 
β-OCH3 [see pdf for image]   03-95   285.06   284.71H NMR (400 MHz, Methanol-d4) δ 8.09- 7.98 (m, 2H), 7.73 (d, J = 8.7 Hz, 1H), 7.65 (t, J = 7.3 Hz, 1H), 7.56 (t, J = 7.6 Hz, 2H), 7.30 (d, J = 2.5 Hz, 1H), 6.96 (dd, J = 8.6, 2.4 Hz, 1H), 3.86 (s, 3H).
 
γ-OCH3 [see pdf for image]   03-98   285.06   284.51H NMR (400 MHz, DMSO-d6) δ 8.08 (d, J = 7.8 Hz, 2H), 7.68-7.60 (m, 2H), 7.59-7.49 (m, 3H), 7.04 (dd, J = 8.8, 2.5 Hz, 1H), 3.80 (s, 3H).
 
  β-OH [see pdf for image]    03-102   271.05   270.8   1H NMR (400 MHz, DMSO-d6) δ 8.12 (d, J = 7.7 Hz, 2H), 7.74 (d, J = 8.6 Hz, 1H), 7.64 (t, J = 7.3 Hz, 1H), 7.55 (t, J = 7.6 Hz, 2H), 7.12 (s, 1H), 6.82 (dd, J = 8.6, 2.3 Hz, 1H).
 
  γ-OH [see pdf for image]   03-60   271.05   271.01H NMR (400 MHz, Methanol-d4) δ 8.05- 7.99 (m, 2H), 7.69-7.51 (m, 4H), 7.25 (d, J = 2.4 Hz, 1H), 6.94 (dd, J = 8.8, 2.5 Hz, 1H)
 
  γ-CN [see pdf for image]    03-106   280.05   279.61H NMR (400 MHz, DMSO-d6) δ 8.46 (s, 1H), 8.14 (d, J = 7.4 Hz, 2H), 7.77 (s, 2H), 7.65-7.45 (m, 3H).
 
  γ-F [see pdf for image]   03-92   273.04   272.61H NMR (400 MHz, DMSO-d6) δ 8.11 (d, J = 7.6 Hz, 2H), 7.97-7.88 (m, 1H), 7.78 (dd, J = 9.1, 4.8 Hz, 1H), 7.65 (t, J = 7.4 Hz, 1H), 7.55 (t, J = 7.6 Hz, 2H), 7.30 (t, J = 9.3 Hz, 1H).
 
γ-CF3 [see pdf for image]    03-105   381.05   380.71H NMR (400 MHz, DMSO-d6) δ 8.72 (s, 1H), 8.53 (s, 1H), 8.39 (d, J = 7.9 Hz, 1H), 8.21 (d, J = 7.8 Hz, 1H), 7.94 (d, J = 8.5 Hz, 1H), 7.81-7.68 (m, 2H), 3.91 (s, 3H).
 
β-OCH3 [see pdf for image]    03-108   343.07   342.81H NMR (400 MHz, DMSO-d6) δ 8.72 (s, 1H), 8.37 (d, J = 7.7 Hz, 1H), 8.14 (d, J = 7.5 Hz, 1H), 7.78 (d, J = 8.6 Hz, 1H), 7.66 (t, J = 7.7 Hz, 1H), 7.21 (s, 1H), 6.89 (d, J = 8.6 Hz, 1H), 3.89 (s, 3H), 3.81 (s, 3H).
 
  γ-F [see pdf for image]   03-110   331.05   330.71H NMR (400 MHz, DMSO-d6) δ 8.73 (s, 1H), 8.39 (d, J = 7.8 Hz, 1H), 8.16 (d, J = 7.7 Hz, 1H), 7.85 (dd, J = 8.7, 2.7 Hz, 1H), 7.70 (dq, J = 11.5, 7.7, 6.2 Hz, 2H), 7.26 (td, J = 9.1, 2.7 Hz, 1H), 3.91 (s, 3H).
 
β-OCH3 [see pdf for image]    03-113   328.07   327.8   1H NMR (400 MHz, DMSO-d6) δ 8.63 (s, 1H), 8.23 (d, J = 7.8 Hz, 1H), 8.11 (s, 1H), 8.07 (d, J = 7.9 Hz, 1H), 7.80 (s, 1H), 7.60 (t, J = 7.8 Hz, 1H), 7.50 (s, 1H), 7.24 (s, 1H), 6.92 (s, 1H), 3.82 (s, 3H).
 
β-OCH3 [see pdf for image]   MS: LY-2-77   342.09   341.81H-NMR (300 MHz; DMSO-d6): δ 8.11 (d, J = 8.7 Hz, 2H), 7.86 (dd, J = 8.7, 0.1 Hz, 1H), 7.75 (d, J = 8.7 Hz, 2H), 7.29- 7.28 (m, 1H), 6.97 (dd, J = 8.7, 2.4 Hz, 1H), 3.84 (s, 3H), 2.10 (s, 3H)
 
  γ-OH [see pdf for image]   08-39   328.07   328.1  
 
  β-OH [see pdf for image]   MS: LY-2-89   328.07   327.81H-NMR (400 MHz; DMSO-d6): δ 8.30 (s, 1H), 7.84-7.82 (m, 2H), 7.75 (d, J = 8.6 Hz, 1H), 7.47 (t, J = 7.9 Hz, 1H), 7.13 (s, 1H), 6.83 (d, J = 8.6 Hz, 1H), 2.09 (s, 3H)
 
  γ-OH [see pdf for image]   03-58   328.07   327.91H NMR (400 MHz, DMSO-d6) δ 8.24 (s, 1H), 7.79 (d, J = 7.2 Hz, 2H), 7.56 (d, J = 8.7 Hz, 1H), 7.44 (t, J = 8.0 Hz, 1H), 7.30 (d, J = 2.5 Hz, 1H), 6.90 (dd, J = 8.6, 2.5 Hz, 1H), 2.06 (s, 3H).
 
β-OCH3 [see pdf for image]   MS. LY-2-85   342.09   341.91H-NMR (300 MHz; DMSO-d6): δ 8.30 (s, 1H), 7.85 (m, 3H), 7.47 (t, J = 7.8 Hz, 1H), 7.29 (d, J = 2.3 Hz, 1H), 6.98 (dd, J = 8.7, 2.3 Hz, 1H), 3.84 (s, 3H), 2.09 (s, 3H).
 
  β-OH [see pdf for image]   MS: LY-2-87   328.07   327.81H-NMR (400 MHz; DMSO-d6): δ 8.10 (d, J = 8.7 Hz, 2H), 7.74 (d, J = 8.6 Hz, 3H), 7.12 (d, J = 2.2 Hz, 1H), 6.82 (dd, J = 8.7, 2.2 Hz, 1H), 2.10 (s, 3H)
 
  γ-OH [see pdf for image]   03-74   328.07   328.31H NMR (400 MHz, Methanol-d4) δ 8.00 (d, J = 8.4 Hz, 2H), 7.72 (d, J = 8.3 Hz, 2H), 7.54 (d, J = 8.7 Hz, 1H), 7.22 (s, 1H), 6.9 (d, J = 8.4 Hz, 1H), 2.13 (s, 3H).
 
γ-CF3 [see pdf for image]   03-80   332.06   331.91H NMR (400 MHz, Methanol-d4) δ 8.24 (s, 1H), 7.87 (d, J = 8.6 Hz, 1H), 7.69 (d, J = 8.6 Hz, 1H), 4.59 (q, J = 7.2, 6.5 Hz, 1H), 2.02 (s, 3H), 1.46 (d, J = 7.2, Hz, 3H).
 

Example 3

Comparative Example

N-(3-acetamidophenyl)-6-hydroxybenzo[d]thiazole-2-carboxamide

[0095]  [see pdf for image]

Step 1. 6-Methoxybenzo[d]thiazol-2-carboxylic acid

[0096] 6-Methoxybenzo[d]thiazole-2-carbonitrile (1 mmol) and concentrated HCl (5 ml) was placed in a sealed microwave vial and heated at 90° C. for one hour with the microwave set for a high-absorbance mixture. The mixture was then diluted with water and the product was isolated by vacuum evaporation.

Step 2. N-(3-acetamidophenyl)-6-methoxybenzo[d]thiazole-2-carboxamide

[0097] 3-Acetamidoaniline (1 eq), 6-methoxybenzo[d]thiazol-2-carboxylic acid (1.05 mmol) and DIPEA (5 mmol) were dissolved in DMF (2 ml). HBTU (1.2 mmol) was added and the reaction were stirred overnight at ambient temperature before extraction with ethyl acetate and washed thoroughly dilute acid and dried. The crude product was isolated by evaporation under vacuum.

Step 3. N-(3-acetamidophenyl)-6-hydroxybenzo[d]thiazole-2-carboxamide

[0098] Borontribromide (2 ml) was added to a suspension of N-(3-acetamidophenyl)-6-methoxybenzo[d]thiazole-2-carboxamide (0.18 mmol) in dichlorormethane at 0° C. before overnight stirring at ambient temperature. The reaction mixture was carefully quenched with water before extraction with ethyl acetate. The crude product was isolated by evaporation under vacuum.
[0099] In a similar manner the following products were prepared:
[00005] [TABLE-US-00005]
 
[see pdf for image]
 
        Calcd mass   Obsd mass   1H NMR
  X   R   Compound   <MH+>   <MH+>   (Exchangeable protons not reported)
 
  γ-OH [see pdf for image]   03-07   328.37   328.01H NMR (400 MHz, DMSO-d6) δ 8.16 (d, J = 2.1 Hz, 1H), 7.99 (d, J = 8.9 Hz, 1H), 7.48 (d, J = 2.4 Hz, 1H), 7.44 (d, J = 8.1 Hz, 1H), 7.39 (d, J = 8.1 Hz, 1H), 7.25 (t, J = 8.1 Hz, 1H), 7.10 (dd, J = 9.0, 2.4 Hz, 1H), 2.03 (s, 3H).
 
γ-OCH3 [see pdf for image]   03-17   342.09   342.11H NMR (400 MHz, DMSO-d6) δ 8.07 (d, J = 9.1 Hz, 1H), 7.80 (d, J = 8.6 Hz, 3H), 7.56 (d, J = 8.5 Hz, 2H), 7.24 (dd, J = 8.8, 2.7 Hz, 1H), 3.88 (s, 3H), 2.03 (s, 3H).
 
γ-OCH3 [see pdf for image]   13-03   285.06   285.01H NMR (400 MHz, DMSO-d6) δ 8.08 (d, J = 9.0 Hz, 1H), 7.88 (d, J = 8.1 Hz, 2H), 7.81 (d, J = 2.6 Hz, 1H), 7.37 (d, J = 7.8 Hz, 2H), 7.24 (dd, J = 9.1, 2.6 Hz, 1H), 7.14
            (t, J = 7.4 Hz, 1H), 3.87 (s, 3H).
 

Example 4

1-(6-fluorobenzo[d]thiazol-2-yl)-3-phenylurea

[0100]  [see pdf for image]
[0101] Phenyl isocyanate (107 mg, 0.9 mmol) was added to a solution of 2-amino-6-fluorobenzothiazole (100 mg, 0.6 mmol) in dry DCM under argon. The reaction was allowed to proceed at R.T. for 16 hours before the product was isolated by filtration.
[0102] In a similar manner the following product was prepared:
[00006] [TABLE-US-00006]
 
[see pdf for image]
 
        Calcd   Obsd  
        mass   mass   1H NMR
  X   R   Compound   <MH+>   <MH+>   (Exchangeable protons not reported)
 
  β-OH [see pdf for image]   13-62   316.07   315.91H NMR (400 MHz, DMSO-d6) δ 7.61 (d, J = 8.5 Hz, 1H), 7.21 (d, J = 9.2 Hz, 2H), 7.01 (d, J = 8.0 Hz, 2H), 6.71 (d, J = 8.3 Hz, 1H), 6.61 (d, J = 8.1 Hz, 1H), 3.76 (s, 3H).
 

B. Kinase Activity

1. Profiling Kinase Inhibitory Activity

DYRK 1A Assay.

[0103] DYRK 1A (5-20 mU of diluted in 50 mM Tris pH7.5, 0.1 mM EGTA) is assayed against Woodtide (KKISGRLSPIMTEQ) (SEQ ID NO. 1) in a final volume of 25.5 μl containing 50 mM Tris pH 7.5, 0.1 mM EGTA, 350 μM substrate peptide, 10 mM Magnesium acetate and 0.05 mM [33P-g-ATP](50-1000 cpm/pmole) and incubated for 30 min at room temperature. Assays are stopped by addition of 5 μl of 0.5 M (3%) orthophosphoric acid and then harvested onto P81 Unifilter plates with a wash buffer of 50 mM orthophosphoric acid.
[0104] Similar assays were performed for the other kinases in the tables which follow using the ADP-Glo™ kinase assay of Promega (products V9101, V9102, V9103 and V9 104).

Results

[0105] Data showing residual kinase activity at 100 micromolar inhibitor concentration is given in the tables which follows. The values are on an absolute scale such that 100 is no change in the presence of putatitic inhibitor, any value over 100 shows kinase activation. Any value less that 25 is considered to be highly significant inhibition.

Results for Compounds of Example 1 (Except 03-20 and 08-70)

[0106] 
[00007] [TABLE-US-00007]
 
  Kinase   01-71   01-66   01-90   03-73   01-94   03-76   03-77   03-32   03-21   03-94   13-10
 
 
  MKK1   35   31   80   96   34   73   87   81   64   76   62
  MKK2   48   42   73   98   54   155   101   96   90
  MKK6   78   70   85   120   98   113   118   85   117
  ERK1   65   61   100   83   97   91   90   83   87
  ERK2   83   104   102   108   91   96   99   149   109
  ERK5         67     62   38   66   57
  JNK1   72   53   104   120   90   97   80   97   97   82   82
  JNK2   82   63   87   111   82   105   99   105   92
  JNK3   66   37   76   83   53   76   68   66   80
  p38a   86   84   106   92   102   92   88   91   96   84   103
  MAPK
  p38b   106   92   82   97   100   83   79   107   101
  MAPK
  p38g   84   46   101   93   82   92   77   109   56
  MAPK
  p38d   43   70   76   52   69   85   76   80   75
  MAPK
  RSK1   25   44   74   89   51   78   72   86   93   85   89
  RSK2   48   66   92   83   87   96   83   105   94
  PDK1   63   69   93   105   97   93   91   96   115   87   93
  PKBa   87   87   104   106   95   104   95   104   91   94   101
  PKBb   57   55   100   104   98   91   93   112   92
  SGK1   33   77   69   77   40   65   66   92   82   67   58
  S6K1   23   54   85   77   73   77   48   92   63   67   65
  PKA   70   95   87   124   93   105   95   91   11   93   99
  ROCK 2   34   56   80   83   69   79   37   91   52   70   96
  PRK2   36   67   94   91   70   84   74   81   67   80   93
  PKCa   77   82   101   111   93   94   101   104   101   98   93
  PKCγ   76   99   53   106   83   106   102   94   103
  PKCz   74   98   81   97   80   101   110   126   85
  PKD1   37   30   119   83   97   89   66   105   59   105   82
  STK33   41   52   46   86   35   80   70   104   81
  MSK1   31   62   87   91   81   84   74   98   93   88   92
  MNK1   28   17   65   89   47   81   64   89   77
  MNK2   34   33   71   96   50   81   86   91   84
  MAPKAP-   46   77   90   114   65   105   106   102   108
  K2
  MAPKAP-   34   39   101   93   101   106   107   61   97
  K3
  PRAK   47   58   111   87   69   75   76   78   71
  CAMKKb   57   44   29   65   35   71   74   101   87   67   73
  CAMK1   52   69   64   89   80   90   106   99   81   86   84
  SmMLCK   39   58   80   71   54   62   66   81   50   67   58
  PHK   47   49   52   91   63   87   89   94   111
  DAPK1   21   44   85   93   41   82   82   89   79
  CHK1   82   75   97   110   96   102   103   105   110
  CHK2   17   31   43   78   30   65   63   87   58   74   59
  CDK2-   30   71   22   96   11   72   66   76   93
  Cyclin A
  CDK9-   48   85   26   97   15   49   46   45   90
  Cyclin
  T1
  PLK1   97   103   80   91   108   90   78   62   90   93   96
  Aurora A   68   87   81   96   61   109   100   109   103
  TLK1   45   92   82   103   53   106   89   92   97
  LKB1   17   38   79   98   35   96   85   97   93   46   73
  AMPK   37   61   90   75   73   77   73   107   100
  AMPK       74   62   40   69   80   86   81   59   70
  (hum)
  MARK1   33   56   90   88   60   94   80   77   98
  MARK2   54   63   74   99   53   97   79   88   103
  MARK3   46   34   61   63   50   71   58   85   72   96   85
  MARK4   66   80   66   97   51   80   86   91   100
  BRSK1   16   48   83   83   62   75   78   95   85
  BRSK2   22   56   64   98   67   79   77   91   95
  MELK   24   17   49   91   34   58   67   83   44
  NUAK1   26   44   66   74   46   58   66   74   84
  SIK2   32   56   72   91   70   71   64   81   89
  SIK3   46   57   82   103   73   103   89   89   121
  TSSK1   67   65   90   71   55   79   84   83   75
  CK1γ2   10   6   59   79   18   54   22   41   6
  CK2   23   54   92   105   66   75   81   96   87   91   51
  TTBK1   61   68   88   94   73   84   80   80   93
  TTBK2       98   91   78   97   93   85   84
  DYRK1A   20   5   8   22   9   23   7   43   2   12   6
  NEK2a   75   85   78   94   60   95   90   90   93
  NEK6   75   82   107   119   86   107   96   92   108   92   92
  IKKb   63   62   75   94   58   59   61   75   98
  IKKe   58   68   81   93   64   89   91   109   91
  TBK1   43   60   83   64   41   78   74   88   81   91   89
  PIM1   19   12   71   10   26   21   19   79   8   40   81
  PIM2   38   16   94   63   73   82   62   100   48
  SRPK1   44   59   41   72   47   73   65   59   84   75   62
  EF2K   75   76   103   109   98   110   99   89   105   96   95
  EIF2AK3   77   59   105   100   78   97   99   104   82
  HIPK1   76   97   48   74   37   53   40   62   39
  HIPK2   14   20   16   51   23   51   22   51   25   27   8
  HIPK3   27   37   69   106   38   79   62   78   86
  PAK2   102   93   103   85   101   102   11   90   95
  PAK4   51   66   67   71   34   101   97   75   94   66   86
  PAK5   85   72   82   85   70   107   111   74   110
  PAK6   81   90   106   11   92   108   106   92   108
  MST2   56   69   99   80   56   75   80   88   91   91   97
  MST3   66   79   85   89   71   85   87   87   91
  MST4   70   84   89   91   75   103   88   110   88
  GCK   19   32   15   36   11   35   8   39   3
  MAP4K3       65   63   52   80   55   90   75
  MAP4K5       40   73   38   83   65   95   96
  MINK1   28   48   35   77   40   63   65   92   61
  MEKK1   96   94   90   111   94   107   102   93   113
  MLK1   11   39   39   75   34   57   58   72   84
  MLK3   15   41   51   57   55   35   44   71   59   46   54
  TESK1   61   94   93   115   90   106   106   95   104
  TAO1   29   30   71   88   46   76   69   73   72
  ASK1   53   70   35   83   18   23   54   66   97
  TAK1   11   30   83   45   17   52   46   76   50   26   58
  IRAK1   26   29   43   85   11   59   46   65   45
  IRAK4   13   18   12   60   10   51   43   65   52   44   8
  RIPK2   22   29   23   48   17   60   64   70   29   38   27
  OSR1   78   95   75   113   79   116   109   112   119
  TTK   31   39   49   86   33   49   64   71   67   37   55
  MPSK1   77   80   105   97   94   91   86   86   97
  WNK1   96   101   100   128   101   104   99   98   121
  ULK1       87   91   92   82   83   102   103
  ULK2       78   88   68   88   83   92   86
  TGFBR1         118     98   96   92   124
  Src   57   45   63   74   94   79   80   113   86   76   85
  Lck   192   69   53   101   168   120   96   91   109   92   79
  CSK   78   92   61   91   83   101   98   83   104
  YES1   59   72   57   108   86   104   85   93   93
  ABL   17   29   31   109   31   82   86   86   111
  BTK   69   67   73   93   98   92   105   100   102   107   96
  JAK2   43   65   73   96   33   51   57   66   88   49   91
  SYK   71   82   100   74   86   72   82   116   72   96   97
  ZAP70   126   113   122   115   118   110   109   111   98
  TIE2   65   75   60   92   85   95   112   112   91
  BRK   22   77   99   106   100   80   91   89   62
  EPH-   66   67   67   99   73   93   106   111   89   94   65
  A2
  EPH-   77   73   45   103   62   91   100   111   98
  A4
  EPH-   86   93   91   114   94   103   103   97   99
  B1
  EPH-   59   70   38   98   42   108   114   116   96
  B2
  EPH-   66   83   89   114   72   107   113   71   106
  B3
  EPH-   98   83   55   101   62   109   112   99   98
  B4
  FGF-R1   42   67   40   109   51   106   103   12   104
  HER4   36   40   97   110   90   85   92   78   94   101   81
  IGF-1R   18   26   84   77   78   75   58   74   78   92   91
  IR   35   63   84   102   81   97   87   87   109
  IRR   34   30   49   61   42   66   57   63   47
  TrkA   42   57   45   74   87   56   59   77   78   58   98
  DDR2   56   65   62   111   93   110   101   97   102
  VEG-   9   24   8   33   11   50   48   66   71   18   5
  FR
  PDGFRA       30   34   40   59   59   78   87
 

Results for Compounds of Example 2

[0107] 
[00008] [TABLE-US-00008]
 
      08-   03-   03-   03-   03-   03-   03-   03-   03-   03-   03-   03-   03-   03-   03-   03-
  Kinase   08-42   44   12   11   33   13   99   95   98   102   60   106   92   105   108   110   113
 
  MKK1   87   67   80   88   89   107   87   92   105   52   49   58   93   86   94   70   119
  MKK2   72   48   82   98   85   82           90
  MKK6   101   107   99   101   122   105           109
  ERK1   84   81   74   88   93   97           67
  ERK2   73   75   88   85   107   100           82
  ERK5   41   43   53   58   58   51           30
  JNK1   99   102   105   95   111   93   86   90   95   83   88   84   94   84   93   98   86
  JNK2   90   90   101   93   92   103           107
  JNK3   65   64   70   77   76   67           48
  p38a   99   108   107   106   105   96   86   82   86   57   74   60   114   84   94   106   113
  MAPK
  p38b   86   89   105   97   87   91           76
  MAPK
  p38g   87   77   89   93   85   94           58
  MAPK
  p38d   65   62   77   82   103   99           75
  MAPK
  RSK1   47   41   67   83   73   70   92   64   74   41   28   96   79   87   70   70   87
  RSK2   62   56   53   71   85   72           34
  PDK1   83   77   88   100   100   104   84   89   101   74   98   106   91   87   101   123   101
  PKBa   90   83   74   88   100   107   91   80   103   88   79   67   99   92   106   109   89
  PKBb   52   45   61   82   93   60           86
  SGK1   60   51   88   84   86   74   69   60   78   86   41   77   93   86   83   87   88
  S6K1   38   17   50   83   75   71   86   77   76   42   11   78   79   78   95   91   79
  PKA   91   92   97   110   102   102   96   83   90   93   97   91   96   96   92   93   101
  ROCK 2   43   42   68   80   53   96   97   54   66   47   11   125   95   103   52   86   68
  PRK2   44   46   66   77   85   97   85   84   91   68   27   105   95   109   100   85   98
  PKCa   81   67   101   91   105   112   114   53   89   72   97   100   96   96   85   109   103
  PKCγ   93   73   143   101   103   141           83
  PKCz   76   81   88   86   105   102           80
  PKD1   63   54   70   74   71   76   114   92   104   60   47   86   111   90   102   88   110
  STK33   67   57   65   77   70   71           38
  MSK1   92   49   90   97   92   100   70   67   90   72   31   64   81   84   88   77   65
  MNK1   53   19   67   45   21   100           18
  MNK2   33   36   66   75   36   66           25
  MAPKAP-   72   56   43   82   109   98           59
  K2
  MAPKAP-   51   38   31   68   97   62           92
  K3
  PRAK   74   60   65   86   83   90           39
  CAMKKb   68   59   41   88   69   69   86   54   45   32   37   67   80   48   80   62   70
  CAMK1   68   42   47   85   104   111   84   76   68   72   22   69   97   92   96   99   102
  SmMLCK   33   19   9   63   52   37   89   48   75   15   15   94   68   63   79   55   83
  PHK   58   43   37   63   74   28           33
  DAPK1   32   26   25   62   82   58           19
  CHK1   70   78   90   78   106   88           72
  CHK2   22   16   23   93   46   75   94   98   37   26   5   62   80   89   65   51   98
  CDK2-   72   72   87   98   92   91           78
  Cyclin A
  CDK9-   66   43   76   68   97   93           76
  Cyclin T1
  PLK1   74   63   65   76   80   96   112   77   102   83   74   103   92   101   96   108   105
  Aurora A   103   99   95   106   108   93           82
  TLK1   97   90   96   95   97   99           74
  LKB1   65   45   64   87   83   45   103   72   118   33   68   107   100   91   90   92   95
  AMPK   61   48   86   84   91   80           30
  AMPK   65   55   90   98   87   71   73   75   94   49   27   80   76   71   101   69   96
  (hum)
  MARK1   75   83   89   90   103   99           96
  MARK2   67   74   90   79   102   84           65
  MARK3   68   65   79   75   85   68   106   106   72   63   33   98   109   103   100   105   101
  MARK4   82   77   90   102   95   71           67
  BRSK1   56   45   63   70   83   95           43
  BRSK2   74   52   79   81   82   80           34
  MELK   22   29   31   48   29   22           18
  NUAK1   57   39   52   86   68   64           14
  SIK2   82   68   77   79   69   96           26
  SIK3   67   80   82   99   92   96           42
  TSSK1   85   109   66   97   75   95           27
  CK1γ2   62   20   57   81   25   83           3
  CK2   50   19   57   68   88   78   80   78   88   75   16   12   90   89   97   86   46
  TTBK1   70   47   91   99   89   100           46
  TTBK2   74   51   102   99   102   71           44
  DYRK1A   15   8   33   50   29   25   9   30   6   6   2   3   11   2   51   13   51
  NEK2a   79   74   92   93   99   89           64
  NEK6   75   84   98   72   107   107   94   86   94   112   97   97   106   90   93   87   99
  IKKb   82   63   84   80   81   88           87
  IKKe   47   50   78   69   81   63           27
  TBK1   61   81   68   80   97   73   96   100   95   66   36   94   92   101   102   96   98
  PIM1   59   42   51   74   25   39   78   19   76   14   35   32   69   72   95   99   50
  PIM2   90   76   75   89   69   77           67
  SRPK1   19   16   63   66   58   57   61   42   10   10   3   7   19   54   68   55   82
  EF2K   79   67   69   79   106   94   99   115   108   92   107   100   134   108   91   94   127
  EIF2AK3   59   51   73   65   87   89           45
  HIPK1   44   43   47   75   41   76           9
  HIPK2   23   19   18   67   52   43   90   74   41   36   8   62   78   72   87   66   96
  HIPK3   90   45   84   106   102   82           38
  PAK2   108   85   104   88   114   119           94
  PAK4   69   64   87   73   92   115   154   107   37   67   35   94   111   89   100   83   109
  PAK5   79   88   94   92   109   82           70
  PAK6   93   94   101   90   119   99           104
  MST2   55   57   61   84   73   60   93   95   86   46   40   103   110   99   77   88   119
  MST3   77   76   85   92   90   78           54
  MST4   79   80   80   134   103   76           71
  GCK   27   31   29   33   25   39           4
  MAP4K3   48   56   54   73   69   67           56
  MAP4K5   67   81   74   92   58   50           31
  MINK1   37   19   48   63   51   31           7
  MEKK1   76   107   127   106   107   114           105
  MLK1   33   13   31   71   61   68           5
  MLK3   30   26   46   95   48   57   72   40   20   15   5   56   49   54   76   68   73
  TESK1   73   70   84   88   102   93           65
  TAO1   81   68   74   77   87   80           62
  ASK1   37   18   70   79   51   88           12
  TAK1   25   18   33   55   73   34   95   45   60   31   9   37   79   76   81   68   35
  IRAK1   38   37   37   68   57   38           26
  IRAK4   42   32   54   69   42   55   70   57   46   34   22   70   72   78   63   96   58
  RIPK2   30   21   29   60   20   27   89   40   34   24   12   78   69   79   39   48   99
  OSR1   72   51   59   90   109   97           82
  TTK   32   22   36   74   45   53   32   71   18   30   19   42   47   9   94   35   95
  MPSK1   77   81   90   73   101   102           106
  WNK1   89   99   91   88   103   100           108
  ULK1   69   78   96   90   88   78           47
  ULK2   63   84   87   86   85   56           69
  TGFBR1   106   103   117   107   88   109           93
  Src   62   72   105   79   75   70   87   77   172   30   33   85   74   165   169   178   79
  Lck   79   82   82   93   105   77   92   93   93   98   92   95   102   99   100   93   111
  CSK   89   83   101   94   88   95           69
  YES1   79   74   73   82   91   60           65
  ABL   76   58   113   99   105   71           54
  BTK   59   54   71   107   110   90   79   75   56   81   59   49   73   95   68   77   74
  JAK2   50   30   71   66   70   98   87   86   81   47   36   87   109   101   85   89   113
  SYK   90   71   99   75   86   106   120   83   109   80   64   93   117   102   96   79   108
  ZAP70   92   79   91   89   110   105           109
  TIE2   80   81   76   84   100   106           41
  BRK   43   21   61   93   46   94           14
  EPH-A2   77   81   85   84   108   81   110   91   95   65   77   89   98   100   76   84   96
  EPH-A4   92   89   78   91   92   89           87
  EPH-B1   93   93   94   99   104   98           98
  EPH-B2   99   91   103   89   99   72           78
  EPH-B3   88   65   63   76   108   44           93
  EPH-B4   92   100   84   97   106   88           110
  FGF-R1   54   56   16   74   102   81           84
  HER4   40   17   33   96   48   98   85   81   73   45   16   82   90   122   99   107   103
  IGF-1R   88   54   54   107   50   84   85   100   104   52   18   131   88   101   123   96   104
  IR   51   35   27   106   105   74           31
  IRR   71   45   82   86   51   78           31
  TrkA   81   71   49   87   51   57   84   35   60   38   23   66   74   82   85   111   87
  DDR2   89   93   104   115   109   85           112
  VEG-FR   20   21   17   58   50   24   28   65   37   11   5   36   53   29   77   66   86
  PDGFRA   40   44   56   63   61   43           29
  PINK
 
      MS:LY-     MS:LY-     MS:LY-   MS:LY-    
    Kinase   2-77   08-39   2-89   03-58   2-85   2-87   03-74   03-80
   
    MKK1   95   106   56   52   83   69   35   80
    MKK2   95   78   47   86   80   73     75
    MKK6   111   96   98   103   99   107     115
    ERK1   87   18   54   84   86   70     94
    ERK2   105   68   68   104   98   81     101
    ERK5   74   47   32   31   45   54     27
    JNK1   104   92   68   80   97   105   80   86
    JNK2   99   93   93   99   100   105     87
    JNK3   88   53   36   51   62   66     79
    p38a   109   96   18   81   89   73   106   79
    MAPK
    p38b   110   96   17   75   92   82     77
    MAPK
    p38g   121   90   64   64   80   93     83
    MAPK
    p38d   98   67   70   75   72   75     102
    MAPK
    RSK1   85   30   13   35   54   29   19   90
    RSK2   81   2   8   51   32   28     66
    PDK1   103   66   44   96   80   69   93   86
    PKBa   89   84   82   92   90   86   87   105
    PKBb   44   4   7   80   22   18     93
    SGK1   101   35   28   41   75   43   59   55
    S6K1   94   12   14   9   50   39   29   97
    PKA   102   88   87   90   98   93   90   104
    ROCK 2   96   22   11   10   29   38   32   80
    PRK2   92   42   44   36   66   67   54   79
    PKCa   97   43   48   96   63   59   92   125
    PKCγ   94   44   63   86   80   74     87
    PKCz   107   38   80   84   101   83     75
    PKD1   114   32   22   54   49   52   53   82
    STK33   99   61   37   45   75   51     60
    MSK1   107   56   46   46   69   66   63   70
    MNK1   85   66   27   33   40   19     78
    MNK2   79   61   40   33   53   43     75
    MAPKAP-   92   56   37   63   69   55     71
    K2
    MAPKAP-   54   9   11   91   24   44     76
    K3
    PRAK   76   17   19   42   30   46     75
    CAMKKb   77   57   22   31   32   38   65   49
    CAMK1   60   8   43   30   62   65   35   50
    SmMLCK   56   34   7   13   13   20   12   40
    PHK   91   72   14   48   40   13     42
    DAPK1   95   31   22   24   48   36     63
    CHK1   107   83   81   79   134   81     75
    CHK2   75   18   10   4   25   17   6   50
    CDK2-   95   75   45   85   106   79     95
    Cyclin A
    CDK9-   88   69   62   86   72   87     6
    Cyclin T1
    PLK1   109   83   61   69   75   68   82   77
    Aurora A   114   97   40   96   103   71     102
    TLK1   102   98   88   78   93   94     105
    LKB1   99   46   28   60   76   104   80   86
    AMPK   95   25   40   26   57   45     82
    AMPK   100   44   37   36   49   42   58   69
    (hum)
    MARK1   96   77   64   69   77   94     58
    MARK2   109   56   27   87   73   50     69
    MARK3   102   48   16   45   58   44   67   78
    MARK4   89   58   49   68   82   59     75
    BRSK1   102   38   38   61   73   35     63
    BRSK2   104   24   37   53   56   48     62
    MELK   59   29   8   13   11   11     83
    NUAK1   52   3   12   37   32   23     80
    SIK2   92   20   7   17   23   36     94
    SIK3   94   40   28   57   77   63     73
    TSSK1   77   39   27   25   35   64     57
    CK1γ2   62   15   28   6   45   57     38
    CK2   103   25   10   23   65   51   7   83
    TTBK1   103   86   84   60   112   68     91
    TTBK2   105   67   73   45   89   73     81
    DYRK1A   55   16   4   4   7   11   4   2
    NEK2a   109   13   64   65   105   76     83
    NEK6   96   36   81   101   97   57   48   81
    IKKb   104   54   57   89   72   71     79
    IKKe   79   14   24   44   51   34     68
    TBK1   89   78   57   50   80   99   72   94
    PIM1   39   26   6   31   9   22   27   65
    PIM2   81   70   19   65   58   42     99
    SRPK1   75   9   4   15   11   16   19   50
    EF2K   111   68   59   103   70   77   95   88
    EIF2AK3   93   67   48   39   61   78     93
    HIPK1   102   68   53   12   84   93     46
    HIPK2   65   17   22   14   39   32   15   8
    HIPK3   107   36   39   48   78   67     25
    PAK2   79   43   59   97   79   84     95
    PAK4   94   66   35   52   51   113   47   74
    PAK5   101   67   57   80   68   120     86
    PAK6   114   92   85   117   94   93     96
    MST2   76   21   19   49   39   44   43   83
    MST3   93   73   55   78   87   68     124
    MST4   85   23   33   60   60   60     76
    GCK   77   26   11   7   25   17     71
    MAP4K3   103   44   10   14   39   33     105
    MAP4K5   80   44   46   37   45   42     86
    MINK1   48   7   3   14   8   5     84
    MEKK1   113   82   103   109   109   90     78
    MLK1   68   13   8   7   28   18     71
    MLK3   91   22   18   7   42   32   6   59
    TESK1   85   69   82   83   74   85     106
    TAO1   99   52   66   77   79   100     87
    ASK1   96   70   46   29   57   49     61
    TAK1   104   23   13   16   37   23   10   90
    IRAK1   78   33   22   29   36   32     73
    IRAK4   83   39   26   33   50   47   21   66
    RIPK2   84   62   12   9   14   26   13   49
    OSR1   96   72   84   86   103   88     95
    TTK   56   25   20   36   31   40   14   56
    MPSK1   104   112   98   103   99   113     97
    WNK1   92   86   110   115   111   111     97
    ULK1   98   80   45   59   55   68     80
    ULK2   115   77   33   61   68   67     93
    TGFBR1   109   110   87   91   124   102     83
    Src   108   54   12   47   210   23   76   96
    Lck   97   79   34   99   76   45   45   90
    CSK   100   84   60   85   85   79     95
    YES1   93   74   14   80   66   21     88
    ABL   108   85   32   64   87   45     101
    BTK   83   37   47   76   100   58   32   98
    JAK2   105   48   32   46   71   47   53   80
    SYK   91   88   78   70   71   75   71   97
    ZAP70   83   103   90   105   118   128     107
    TIE2   103   76   46   59   92   67     91
    BRK   90   59   45   22   63   79     84
    EPH-A2   94   54   63   97   81   69   73   90
    EPH-A4   86   104   49   99   105   86     74
    EPH-B1   101   86   87   106   103   79     103
    EPH-B2   124   108   53   89   130   67     93
    EPH-B3   76   44   45   101   75   67     124
    EPH-B4   83   107   74   100   113   92     97
    FGF-R1   95   16   47   91   89   69     98
    HER4   67   31   20   29   66   32   31   102
    IGF-1R   96   46   61   32   92   80   33   71
    IR   74   49   29   51   67   70     91
    IRR   89   66   68   28   70   70     52
    TrkA   75   56   45   36   73   44   56   97
    DDR2   112   88   62   104   94   76     84
    VEG-FR   90   8   12   7   47   17   11   19
    PDGFRA   90   44   28   32   47   43     38
    PINK                 112
   

Results for Compounds of Example 3

[0108] 
[00009] [TABLE-US-00009]
   
    Kinase   03-07   03-17   13-03
   
 
    MKK1   85   106   70
    MKK2   88   50
    MKK6   114   106
    ERK1   102   115
    ERK2   96   103
    ERK5   48   79
    JNK1   103   103   88
    JNK2   104   87
    JNK3   50   86
    p38a MAPK   93   100   103
    p38b MAPK   96   81
    p38g MAPK   93   95
    p38d MAPK   81   97
    RSK1   48   93   77
    RSK2   71   105
    PDK1   72   104   113
    PKBa   103   96   106
    PKBb   26   114
    SGK1   64   215   91
    S6K1   26   92   90
    PKA   91   98   97
    ROCK 2   27   83   70
    PRK2   74   89   105
    PKCa   81   87   119
    PKCγ   98   116
    PKCz   107   108
    PKD1   61   102   113
    STK33   63   85
    MSK1   62   101   97
    MNK1   40   103
    MNK2   50   85
    MAPKAP-K2   51   95
    MAPKAP-K3   34   76
    PRAK   59   79
    CAMKKb   30   103   91
    CAMK1   44   90   94
    SmMLCK   21   63   85
    PHK   55   106
    DAPK1   28   85
    CHK1   113   86
    CHK2   23   83   92
    CDK2-Cyclin A   99   94
    CDK9-Cyclin T1   76   80
    PLK1   93   94   93
    Aurora A   110   112
    TLK1   108   103
    LKB1   83   96   110
    AMPK   76   86
    AMPK (hum)   34   100   99
    MARK1   106   108
    MARK2   73   90
    MARK3   72   86   92
    MARK4   71   94
    BRSK1   52   109
    BRSK2   66   102
    MELK   12   79
    NUAK1   37   85
    SIK2   28   68
    SIK3   56   75
    TSSK1   57   98
    CK1γ2   77   82
    CK2   35   50   97
    TTBK1   93   92
    TTBK2   71   108
    DYRK1A   30   45   45
    NEK2a   38   85
    NEK6   91   95   107
    IKKb   88   90
    IKKe   73   94
    TBK1   92   82   98
    PIM1   25   74   96
    PIM2   61   82
    SRPK1   27   84   86
    EF2K   91   98   110
    EIF2AK3   69   88
    HIPK1   90   95
    HIPK2   46   73   96
    HIPK3   81   105
    PAK2   90   103
    PAK4   92   92   58
    PAK5   82   77
    PAK6   106   92
    MST2   68   98   107
    MST3   85   91
    MST4   76   105
    GCK   35   84
    MAP4K3   63   88
    MAP4K5   56   85
    MINK1   18   84
    MEKK1   115   120
    MLK1   26   99
    MLK3   39   80   89
    TESK1   101   83
    TAO1   84   113
    ASK1   86   94
    TAK1   53   69   96
    IRAK1   49   98
    IRAK4   74   126   93
    RIPK2   18   96   76
    OSR1   87   97
    TTK   28   91   108
    MPSK1   108   88
    WNK1   115   97
    ULK1   70   85
    ULK2   81   121
    TGFBR1   113   121
    Src   101   102   100
    Lck   88   88   89
    CSK   105   93
    YES1   49   104
    ABL   105   103
    BTK   32   95   31
    JAK2   56   90   93
    SYK   102   82   37
    ZAP70   94   196
    TIE2   78   100
    BRK   80   137
    EPH-A2   88   90   63
    EPH-A4   95   119
    EPH-B1   104   105
    EPH-B2   110   96
    EPH-B3   64   98
    EPH-B4   100   137
    FGF-R1   66   80
    HER4   50   109   130
    IGF-1R   96   118   104
    IR   66   86
    IRR   75   84
    TrkA   59   94   55
    DDR2   90   109
    VEG-FR   28   114   87
    PDGFRA   44   65
   

Results for Compound of Example 4

[0109] 
[00010] [TABLE-US-00010]
   
    Kinase   13-62
   
 
    MKK1   15
    JNK1   114
    p38a MAPK   21
    RSK1   25
    PDK1   77
    PKBa   66
    SGK1   21
    S6K1   22
    PKA   104
    ROCK 2   11
    PRK2   40
    PKCa   102
    PKD1   41
    MSK1   41
    CAMKKb   25
    CAMK1   8
    SmMLCK   5
    CHK2   18
    PLK1   98
    LKB1   91
    AMPK (hum)   23
    MARK3   12
    CK2   24
    DYRK1A   7
    NEK6   104
    TBK1   48
    PIM1   14
    SRPK1   3
    EF2K   85
    HIPK2   23
    PAK4   22
    MST2   3
    MLK3   13
    TAK1   11
    IRAK4   17
    RIPK2   6
    TTK   25
    Src   7
    Lck   11
    BTK   25
    JAK2   31
    SYK   59
    EPH-A2   81
    HER4   13
    IGF-1R   62
    TrkA   9
    VEG-FR   7
   

2. Determination of IC50 Values for Selected Inhibitors from Example 1 Against DYRK1a
[0110] IC 50 values were determined by measuring the inhibition of DYRK1a caused by each compound at a compound concentration of 100 μM, 30 μM, 10 μM, 3 μM, 1 μM, 0.3 μM, 0.1 μM, 0.03 μM, 0.01 μM and 0.003 μM. The kinase assay was carried out using Multidrop 384's at room temperature in a total assay volume of 25.5 μl. To plates containing 0.5 μl of compounds, DMSO controls or acid blanks, 15 μl of an enzyme mix containing the DYRK1a enzyme and peptide/protein substrate in buffer was added. Compounds were pre-incubated in the presence of the enzyme and peptide/protein substrate for 5 minutes before initiation of the reaction by addition of 10 μl of ATP (final concentration selected for each kinase at 5, 20 or 50 μM). Assays were carried out for 30 minutes at room temperature before termination by the addition of 5 μl orthophosphoric acid. The assay plates were then harvested onto P81 Unifilter Plates by a Packard Harvester (wash buffer is 50 mM orthophosphoric acid) and dried in air. The dry Unifilter plates were then sealed by the addition of MicroScint O and counted in Packard Topcount NXT scintillation counters. The measurements were performed in duplicate and the average determined and reported as the IC50 value and presented in the table below.
[00011] [TABLE-US-00011]
  TABLE
 
  Determined IC50 values (in μM) for inhibitors against DYRK1a.
[see pdf for image]
 
  X   Compound   IC 50 (μM)
 
  γ-OH   01-71   28.1 
  β-OH   01-66   0.8
γ-OCH3   03-20   12.3 
β-OCH3   03-21   0.4
  γ-F   03-94   23.6 
  β-F   03-77   3.9
γ-CF3   01-90   1.1
β-CF3   03-73   26.5 
  γ-CN   01-94   1.3
γ-NO2   13-10   0.7
γ-CONH2   08-70   15.7 
 
(57)

Claim

1.-2. (canceled)
3. A method of treating or preventing a neurodegenerative disorder in a subject comprising administering a therapeutically effective amount of a compound of formula (I)
[see pdf for image]
wherein:
Y represents a C or N atom which may be substituted or form a cyclic group with R′″ but may not be a quaternary C atom;
R′ is —OR1, —CONH2, —CF3, F, —OH, —NO2, —CN or —OCOR1 in which R1 is C1-3 alkyl and each may be in the beta or gamma position;
R″ is C1-3 alkyl or H; and
R′″ is H or a group consisting of 1-12 non-hydrogen atoms and may be linear, branched and/or incorporate one or more cyclic groups, cyclic groups may be aromatic and/or heterocyclic and 2 or more cyclic groups may be linked or fused and each may be substituted;
or a salt, hydrate or solvate of a compound of formula (I),
wherein the therapeutically effective amount is such as to inhibit formation of neurofibrillary (tau) tangles.
4. The method of claim 3 wherein Y represents a C atom and forms a cyclic group with R′″.
5. The method of claim 3 wherein R′″ is hydrogen.
6. The of claim 3 wherein Y is —CH2— or —NH—.
7. The method of claim 3 wherein R′″ is a linear or branched, optionally substituted, C1-5 alkyl group or a derivative thereof.
8. The method of claim 3 wherein R′″ is an amino acylated amino acid or an acylated dipeptide.
9. The method of claim 3 wherein R′″ or YR′″ together contain 1 to 3 cyclic groups.
10. The method of claim 9 wherein R′″ or YR′″ together contain a single cyclic, aromatic, optionally substituted group.
11. The method of claim 9 wherein R′″ or YR′″ is or comprises a cyclic group selected from cyclohexyl, phenyl, pyridine, pyrimidine, pyrazole, imidazole, thiazole, oxazole, morpholine and piperidine.
12. The method of claim 9 wherein one or more of the cyclic groups of R′″ or YR′″ is substituted by a group selected from:
methyl, a halogen, hydroxyl, amino and sulfhydryl;
ethyl, ethenyl, ethynyl, methoxy, methylamino, methylsulfide, cyano and formyl;
propyl, isopropyl, cyclopropyl, methylsulfoxy, acetyl, nitro, dimethylamino, CH2CH2OH, CH2CH2NH2, carboxylate and carboxamide;
—OCH2CH2OH, —NHCH2CH2OH, —OCH2CH2NH2, —NHCH2CH2NH2, methylcarboxylate, N-methylcarboxamide, trifluoromethyl, methylsulfonyl, sulphonamide and sulfonic acid;
methylsulfonylamido, trifluoromethoxy, cyclopropylmethoxy and N,N-dimethylcarboxamido;
piperidyl, morpholinyl, —C═ONHCH2CH2NH2, —C═ONHCH2CH2OH and C═OOCH2CH2OH;
N(CH2CH2NH2)2, N(CH2CH2OH)2 and methylpiperidyl.
13. The method of claim 3 wherein R″ is H or methyl.
14. The method of claim 3 wherein the neurodegenerative disorder is selected from the list consisting of Alzheimer's disease, Parkinson's disease, Pick's disease, progressive supranuclear palsy, corticobasal degeneration and agyrophilic grain disease.
15. The method of claim 14 wherein the neurodegenerative disorder is Alzheimer's disease in patients with Downs Syndrome.
16. An in vitro method of inhibiting Dyrk1A or a reaction catalysed by Dyrk1A, the method comprising contacting said kinase with a compound as defined claim 3.
17. (canceled)
*****

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