UBC Faculty Research and Publications

A phase Ib multiple ascending dose study of the safety, tolerability, and central nervous system availability… Nygaard, Haakon B; Wagner, Allison F; Bowen, Garrett S; Good, Susan P; MacAvoy, Martha G; Strittmatter, Kurt A; Kaufman, Adam C; Rosenberg, Brian J; Sekine-Konno, Tomoko; Varma, Pradeep; Chen, Kewei; Koleske, Anthony J; Reiman, Eric M; Strittmatter, Stephen M; van Dyck, Christopher H Apr 14, 2015

Your browser doesn't seem to have a PDF viewer, please download the PDF to view this item.

Item Metadata


52383-13195_2015_Article_119.pdf [ 879.86kB ]
JSON: 52383-1.0223303.json
JSON-LD: 52383-1.0223303-ld.json
RDF/XML (Pretty): 52383-1.0223303-rdf.xml
RDF/JSON: 52383-1.0223303-rdf.json
Turtle: 52383-1.0223303-turtle.txt
N-Triples: 52383-1.0223303-rdf-ntriples.txt
Original Record: 52383-1.0223303-source.json
Full Text

Full Text

RESEARCH Open AccessA phase Ib multiple ascending dose study of thesafety, tolerability, and central nervous systememission tomography.Nygaard et al. Alzheimer's Research & Therapy  (2015) 7:35 DOI 10.1186/s13195-015-0119-0Haven, Connecticut, USAFull list of author information is available at the end of the articleResults: AZD0530 was generally safe and well tolerated across doses. One subject receiving 125 mg of AZD0530 wasdiscontinued from the study due to the development of congestive heart failure and atypical pneumonia, which wereconsidered possibly related to the study drug. Plasma/CSF ratio of AZD0530 was 0.4. The 100 mg and 125 mg dosesachieved CSF drug levels corresponding to brain levels that rescued memory deficits in transgenic mouse models.One-month treatment with AZD0530 had no significant effect on clinical efficacy measures or regional cerebral glucosemetabolism.(Continued on next page)* Correspondence: stephen.strittmatter@yale.edu; christopher.vandyck@yale.edu2Department of Neurology, Yale University School of Medicine, New Haven,Connecticut, USA1Alzheimer’s Disease Research Unit, Yale University School of Medicine, NewScale – Sum of Boxes) and regional cerebral glucose meavailability of AZD0530 (saracatinib) in Alzheimer’sdiseaseHaakon B Nygaard1,2,3,8, Allison F Wagner1,4, Garrett S Bowen1,4, Susan P Good1,4, Martha G MacAvoy1,4,Kurt A Strittmatter3, Adam C Kaufman3, Brian J Rosenberg5, Tomoko Sekine-Konno2, Pradeep Varma6, Kewei Chen7,Anthony J Koleske3,5, Eric M Reiman7, Stephen M Strittmatter2,3* and Christopher H van Dyck1,2,4*AbstractIntroduction: Despite significant progress, a disease-modifying therapy for Alzheimer’s disease (AD) has not yet beendeveloped. Recent findings implicate soluble oligomeric amyloid beta as the most relevant protein conformation in ADpathogenesis. We recently described a signaling cascade whereby oligomeric amyloid beta binds to cellular prionprotein on the neuronal cell surface, activating intracellular Fyn kinase to mediate synaptotoxicity. Fyn kinase hasbeen implicated in AD pathophysiology both in in vitro models and in human subjects, and is a promising newtherapeutic target for AD. Herein, we present a Phase Ib trial of the repurposed investigational drug AZD0530, aSrc family kinase inhibitor specific for Fyn and Src kinase, for the treatment of patients with mild-to-moderate AD.Methods: The study was a 4-week Phase Ib multiple ascending dose, randomized, double-blind, placebo-controlledtrial of AZD0530 in AD patients with Mini-Mental State Examination (MMSE) scores ranging from 16 to 26. A total of 24subjects were recruited in three sequential groups, with each randomized to receive oral AZD0530 at doses of 50 mg,100 mg, 125 mg, or placebo daily for 4 weeks. The drug:placebo ratio was 3:1. Primary endpoints were safety,tolerability, and cerebrospinal fluid (CSF) penetration of AZD0530. Secondary endpoints included changes inclinical efficacy measures (Alzheimer’s Disease Assessment Scale – cognitive subscale, MMSE, Alzheimer’s DiseaseCooperative Study – Activities of Daily Living Inventory, Neuropsychiatric Inventory, and Clinical Dementia Ratingtabolism measured by fluorodeoxyglucose positron© 2015 Nygaard et al.; licensee BioMed Central. This is an Open Access article distributed under the terms of the CreativeCommons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, andreproduction in any medium, provided the original work is properly credited. The Creative Commons Public DomainDedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article,unless otherwise stated.tel dIIaedNygaard et al. Alzheimer's Research & Therapy  (2015) 7:35 Page 2 of 11memory deficits, reduced survival, epileptiform discharges,synapse loss, serotonin fiber degeneration and for celldeath in vitro [10-17]. Human AD brain-derived Aβspecies suppress hippocampal synaptic plasticity in vivoin a PrPC-dependent fashion, and human AD brain ex-tracts contain Aβo species that interact with PrPC aswell as Aβ-PrPC complexes [10,18-20]. Several groups,including ours, recently showed that the Aβo-PrPC com-plex activates an intracellular signaling cascade coupled toStudy protocolThis study protocol was reviewed and approved by theYale Human Investigation Committee. All study partici-pants and their study partners provided written informedconsent to participate in this study. Given the absence ofpotential benefit in this short Phase Ib study, participantswere required to provide informed consent for themselves(that is, surrogate consent was not permitted).(Continued from previous page)Conclusions: AZD0530 is reasonably safe and well tolerasubstantial central nervous system penetration with orapromising therapeutic approach in AD, and a larger PhaseAD has recently launched.Trial registration: ClinicalTrials.gov: NCT01864655. RegisterIntroductionDespite considerable ongoing efforts to halt or reversethe symptoms of Alzheimer’s disease (AD), a disease-modifying intervention for this devastating illness hasnot yet emerged. The major approach to AD therapeuticdevelopment has been to target amyloid-beta (Aβ), byeither limiting its cleavage from the amyloid precursorprotein, or facilitating its clearance by active or passiveimmunization [1]. An alternative approach to AD treat-ment is to target the downstream effects of pathologic Aβsignaling, without altering protein levels. This may be aparticularly attractive approach as some forms of Aβ couldhave important physiologic roles [2,3]. More recently, pre-clinical focus has changed from insoluble assemblies ofAβ to oligomeric species (Aβo) and, while the precise na-ture of the toxic Aβo is not fully understood, a unifyingtheme among various preparations and sources is synap-totoxicity [4-6]. Blocking this detrimental downstreameffect of Aβo may be a key feature of future Aβ-basedtherapeutics in AD.The mechanism by which Aβo causes synaptoxicity isof significant interest, with a leading hypothesis postulat-ing the existence of a distinct cell surface receptor medi-ating its effects. To identify such a potential binding siteof Aβo on neurons, we performed a genome-wide unbiasedscreen for cell surface proteins binding Aβo, identifying cel-lular prion protein (PrPC) as a high-affinity receptor [5].Aβ binding to PrPC is highly conformation-dependent, andonly Aβo will interact with PrPC, with no binding detectedin the presence of fibrillary or monomeric Aβ peptide[5,7-9]. Emphasizing its potential importance, PrPC is re-quired for most deficits seen in amyloid precursor protein/presenilin 1 transgenic mice, including spatial learning andthe protein tyrosine kinase Fyn [5,10,21,22]. This is of par-ticular interest since Fyn has been implicated in AD patho-genesis across various models, including human subjectsd in patients with mild-to-moderate AD, achievingosing at 100–125 mg. Targeting Fyn kinase may be aclinical trial of AZD0530 for the treatment of patients with12 June 2014.[23]. Fyn is central to Aβ signal transduction, and also hasmajor functional interactions with Tau [24-27], therebyunifying the two principal pathologies in AD. Thus, block-ing Fyn kinase may be a promising therapeutic approach inAD.Saracatinib (AZD0530) is a small molecule inhibitor ofthe Src family kinases, blocking Src, Fyn, Yes and Lyn,with 2 to 10 nM potency [28]. At 10- to 100-fold higherconcentrations the compound also inhibits Abl familykinases, without detectable activity in this concentrationrange against other kinase families, including c-kit, Cskand platelet-derived growth factor. Due to its specific in-hibition of Fyn and Src family kinases, and the fact thatSrc family kinases regulate cell proliferation and tumorcell adhesion, migration and invasion, AZD0530 was ini-tially developed as a therapy for solid tumors [28]. Todate, more than 500 subjects have received once-dailyoral doses of AZD0530. Preliminary studies indicate thatthe dose needed to disrupt the Aβ-PrPC-Fyn signalingcascade is significantly lower than that needed for cancertherapy, and doses up to 125 mg daily have been foundto be safe and well tolerated (unpublished data).Herein we present the results of a 4-week multiple as-cending dose study of the safety, tolerability, and centralnervous system (CNS) availability of AZD0530 (saracatinib)in mild-to-moderate AD. We assessed the safety and toler-ability of three escalating doses of AZD0530, as well as theability of the compound to penetrate the CNS, a critical fea-ture for any AD drug. Secondary aims included changes inclinical efficacy measures and in regional cerebral glucosemetabolism.MethodsStudy populationMen and women aged 50 to 90 years old (inclusive) witha diagnosis of probable AD dementia based on theNygaard et al. Alzheimer's Research & Therapy  (2015) 7:35 Page 3 of 11National Institute on Aging-Alzheimer’s Association coreclinical criteria [29], and Mini-Mental State Examination(MMSE) [30] score of 16 to 26 were considered for studyparticipation. Additionally, subjects were required to havestable permitted medications for 4 weeks prior to studyparticipation. Cholinesterase inhibitors and memantinewere required to be stable for 12 weeks prior to screen.Participants also needed to score less than 6 on theGeriatric Depression Scale [31] and less than or equalto 4 on the Modified Hachinski Scale [32]. Additionally,they needed to have completed six grades of educationor have a good work history and speak English fluently.Amyloid positron emission tomography (PET) imagingwas not used to determine study eligibility.Participants were excluded from the study if they ex-hibited any significant neurologic disease other than AD.Additionally, subjects with a screening magnetic resonanceimaging scan showing evidence of infection, infarction, orother focal lesions as well as multiple lacunes or lacunesin a brain region critical for memory were excluded. Ahistory of schizophrenia or of alcohol or substance abuse/dependence within the past 2 years (Diagnostic andStatistical Manual of Mental Disorders, 4th edition criteria)was also exclusionary. Any subject with a clinically signifi-cant or unstable medical condition that could have putthe subject at risk due to study participation, or influ-ence the results, was excluded. Additionally, the followinglaboratory results were considered exclusionary: clinicallysignificant abnormalities in vitamin B12 or thyroid func-tion tests that might interfere with the study, neutropeniadefined as an absolute neutrophil count of <1,500/μl,thrombocytopenia defined as platelet count <100 × 103/μl,current blood clotting or bleeding disorder, or signifi-cantly abnormal prothrombin time or partial thromboplas-tin time at screening, or clinically significant abnormalitiesin other screening laboratories, including aspartate amino-transferase >1.5 × upper limit of normal (ULN); alanineaminotransferase >1.5 × ULN; total bilirubin >1.5 × ULN;serum creatinine >2.0 × ULN. Additionally, a history ofinterstitial lung disease was exclusionary. Residence ina skilled nursing facility was also considered exclusionary.Current use (within 30 days of screening) of the follow-ing medications was exclusionary: typical neuroleptics,narcotic analgesics, Parkinson’s medications, systemiccorticosteroids, or medications with significant centralanticholinergic activity, or warfarin. Current use (within30 days of screening and throughout the protocol in-cluding the 2-week follow-up period) of the followingmedications was exclusionary: a) strong CYP3A4 inhib-itors including: atazanavir, indinavir, ritonavir, saquinavir,nelfinavir, ketoconazole, itraconazole, clarithromycin,telithromycin, and nefazodone; b) strong CYP3A4 in-ducers including: rifampicin, phenytoin, phenobarbital,and carbamazepine; c) certain CYP3A4 substrates includingcolchicine, cyclosporine, disopyramide, fluticasone, quini-dine, vinblastine, vincristine. Subjects taking sildenafil, tada-lafil, and vardenafil were advised to stop taking thesemedications for the duration of the trial.Study designThe study was conducted in the Yale Alzheimer’s DiseaseResearch Unit between July 2013 and March 2014. This4-week, multiple ascending dose, randomized, double-blind, placebo-controlled, Phase Ib trial enrolled 24 subjects.The subjects were divided into three sequential cohorts.Each cohort contained six subjects assigned to active drugand two subjects assigned to placebo. The dose of the firstcohort was 50 mg of oral AZD0530 or placebo per day for 4to 5 weeks. The dose of the second and third cohorts was tobe determined from the results of cohort 1 and ongoingpreclinical studies, not to exceed 125 mg daily. Eachcohort of eight participants completed the study beforethe next cohort was recruited.The primary aims of this study were to assess the safetyand tolerability of oral AZD0530 in patients with AD andto determine dose levels that are well tolerated and pro-vide cerebrospinal fluid (CSF) concentrations predictedto slow AD progression. Secondary aims were to assesseffects of AZD0530 on clinical efficacy assessments andbrain 18F-fluorodeoxyglucose (FDG) PET changes in astatistical region of interest (sROI) known to be prefer-entially affected in AD over longer time periods.Potential participants who were found to be eligiblebased on the initial screening evaluation proceeded to thebaseline assessments. These included safety assessments,clinical efficacy assessments, 18F-FDG PET imaging, col-lection of AZD0530 metabolites and AD biomarkers, anddispensation of study medication. Subjects were seen forsafety visits at weeks 1, 2, and 3. Outcome measures werecollected after 4 weeks on AZD0530 or placebo, includingmeasures of safety and tolerability, CSF drug concen-tration, clinical efficacy measures, and 18F-FDG PETimaging. Study medication could be continued for upto 5 weeks (35 days) in order to complete all outcomemeasures. Study medication compliance was measuredthroughout the study. A final safety visit was completedafter the participant had been off AZD0530 or placebo forapproximately 2 weeks.Safety assessmentsSafety assessments included physical and neurologicalexaminations, MMSE examinations, vital signs (includingblood pressure, pulse, oral temperature, respiration rate,and weight), electrocardiograms, and clinical laboratories.Physical and neurological examinations occurred at allvisits except the baseline visit. Electrocardiograms oc-curred at the screening, and at week 4 and week 6 visits.Vital signs, clinical laboratories, and MMSE examinationsNygaard et al. Alzheimer's Research & Therapy  (2015) 7:35 Page 4 of 11occurred at all visits. Previous clinical experience withAZD0530 has indicated a possible, but rare, relationshipwith interstitial lung disease in patients with advancedsolid tumors [33]. For this reason, thoracic high-resolutioncomputed tomography was obtained if unexplained pul-monary symptoms arose at any point during the study.Clinical efficacy measuresAt the baseline and week 4 visits, subjects underwentthe following clinical efficacy assessments: Alzheimer’sDisease Assessment Scale – cognitive subscale (ADAS-cog)[34], MMSE [30], Alzheimer’s Disease Cooperative Study –Activities of Daily Living Inventory (ADCS-ADL) [35],Neuropsychiatric Inventory (NPI) [36], and ClinicalDementia Rating Scale – Sum of Boxes (CDR-SOB) [37].The MMSE was also completed at screening and weeks 1,2, and 3.18F-FDG PET imagingEligible subjects had 18F-FDG PET brain scans performedat the baseline visit and at week 4. A 30-minute dynamicemission scan, consisting of six 5-minute frames was ac-quired starting 30 minutes after the intravenous injectionof 5 mCi of 18F-FDG. Data was corrected for radiation-attenuation and scatter using transmission scans or X-raycomputed tomography, and reconstructed using a stan-dardized reconstruction algorithm.Peripheral Src family kinase target engagementOsteoclast function is known to be mediated through Src,and therefore levels of cross-linked serum C-telopeptideof type 1 collagen (sCTX) measurements have been usedin several studies as a surrogate marker of relevant bio-logic function of AZD0530 [38,39]. sCTX levels weremeasured at the baseline visit and at week 4 by enzyme-linked immunosorbent assay (Immunodiagnostic Systems,Boldon, UK) according to the manufacturer’s instructions.Serum samples were stored at −80°C until analysis. Allsamples collected from each individual subject were ana-lyzed in the same batch by an investigator blinded to sub-ject drug assignment.Abl inhibition assayAn assay for plasma inhibitory activity against Abl kinasefollowed previous procedures [40,41] by determiningthe level of Abl-dependent phosphorylation of Stat5 inleukemia cells incubated with plasma from patients. TheK-562 cells, a gift from Prof. Gary Kupfer, were main-tained in RPMI 1640 supplemented with 10% fetal bovineserum. To generate a dose–response curve for AZD0530and STI-571, 1 × 105 K-562 cells were incubated in 100 μlnormal volunteer plasma in 96 well plates for 2 hours at37°C in a humidified incubator with 5% CO2, with the ap-propriate amount of drug added, diluted in equal volumesof DMSO. For experiments with patient samples, cellswere incubated in patient plasma samples, with the ex-perimenter blinded to patient group. Cells were pelletedby centrifuge, washed twice in 500 μl phosphate-bufferedsaline, then lysed in 40 μl 2× Laemmli sample buffer(LSB), boiled for 10 minutes and centrifuged. Lysates werethen loaded onto 8% SDS-PAGE gels, transferred tonitrocellulose membranes, and immunoblotted using aphospho-specific Stat5 antibody (Cell Signaling, Danvers,MA, USA), and then stripped and re-probed using a Stat5antibody (SantaCruz, Dallas, TX, USA). Immunoblotswere then quantified by densitometry, and the densityof each band normalized to a control sample treated withvolunteer plasma. Nonlinear regression was performedto generate a dose–response curve of inhibition versusAZD0530 and STI concentration.Cerebrospinal fluid Alzheimer’s disease biomarkersLevels of CSF Aβ40, Aβ42, Tau and P231-Tau were measuredusing a Luminex assay system and a MagPix instrument,according to the manufacturer’s instructions (Millipore,Billerica, MA, USA). All assays were performed in a singlebatch by an investigator blinded to subject assignments.Rodent AZD0530 pharmacokineticsTo examine the correlation between plasma, CSF, andbrain levels of AZD0530 in a preclinical model, five mice(C57B6/J) received a dose of 5 mg/kg per day adminis-tered orally twice daily for 3 days. Trough plasma, CSFand brain drug levels were measured by tandem reverse-phase high-pressure liquid chromatography and massspectrometry. All animal work was approved by the YaleInstitutional Animal Care and Use Committee accordingto established international guidelines.Statistical analysisAt the completion of all three dose cohorts, the frequenciesof adverse events or laboratory abnormalities between theparticipants who received each dose of AZD0530 (n = 6)and the pooled placebo subjects (n = 6) were comparedusing Fisher’s exact test. Changes in clinical outcomes(MMSE, ADAS-cog, CDR-SOB, ADCS-ADL, NPI) wereexamined using analysis of covariance or Kruskal-WallisH test (for non-normally distributed NPI data). Change in18F-FDG PET cerebral metabolic rate for glucose (CMRgl)from baseline to week 4 was analyzed using statistical para-metric mapping sROI methods as previously published [42].The effect of treatment assignment on change in CMRgl wasanalyzed by four-group analysis of variance and t-test.ResultsSubject dispositionThirty-one subjects were screened, with 24 meeting eli-gibility criteria. Seven subjects were excluded as follows:inability to consent (2), MMSE >26 (2), abnormal thy-roid stimulating hormone (1), thalamic infarct (1), andabnormal liver function tests (1). Of the 24 subjects,18/24 were randomized to one of three doses of AZD0350,and the remaining six to placebo. One subject discontinuedtreatment prematurely due to a serious adverse event, asdetailed below.Baseline characteristicsOf the 24 subjects, 61% were female, with age rangingfrom 55 to 86 years (mean ± SD, 73 ± 7) (Table 1); 96%of subjects were Caucasian. Mean score on the MMSEat baseline was 22.2 (±3). The treatment groups did notdiffer significantly on any demographic or baseline clinicalefficacy variable. However, the 125-mg group was slightlyolder and had a lower baseline MMSE score in compari-son to other groups.Safety and tolerabilityIn general, doses of AZD0530 ranging from 50 mg to125 mg daily were reasonably well tolerated in patientswith mild-to-moderate AD. The number of participantsin each AZD0530 treatment group experiencing adverseevents is presented in Table 2. Treatment-emergent ad-verse events were reported for three participants in theplacebo group, and four, five, and four participants inthe 50-mg, 100-mg, and 125-mg groups, respectively. Alladverse events were of mild or moderate severity. Twoparticipants in the 125-mg dose group had moderatelysevere events, compared to one each for the 100-mg andplacebo groups. The most common individual adverseevents were diarrhea, headache, fatigue, and nausea, whichtended to occur more frequently with higher doses ofAZD0530 but were numerically most frequent in theplacebo group. The effect of treatment group was notstatistically significant for any adverse event.The only serious adverse event (SAE) occurred in aparticipant who was receiving 125 mg AZD0530. After9 days of treatment, the subject was hospitalized with4 days of fatigue, anorexia, and myalgias, and 1 day ofshortness of breath. A low-grade fever with a maximumtemperature of 100.1 °F was recorded after hospital ad-mission. The diagnosis was felt to be most consistentwith congestive heart failure, precipitated by pneumonia;however, a drug-induced pneumonitis could not be en-tirely ruled out. An infectious organism was not isolated.Table 1 Subject characteristics and clinical efficacy assessmentsVariable Treatment group (dose of AZD0530)Placebo (n = 6) 50 mg (n = 6) 100 mg (n = 6) 125 mg (n = 6) F PAge 72.6 ± 9.5 72.6 ± 4.4 71.5 ± 4.9 4.6 ± 7.9 0.57 † 0.19areddruNygaard et al. Alzheimer's Research & Therapy  (2015) 7:35 Page 5 of 11Sex 3 F, 3 M 5 F, 1 MEducation (years) 15.0 ± 2.8 15.3 ± 2.7Weight (kg) 6.88 ± 12.1 69.7 ± 8.7MMSE, baseline 23.0 ± 4.1 21.8 ± 3.3MMSE, week 4 25.7 ± 3.3 25.3 ± 2.4MMSE, change 2.7 ± 2.0 3.5 ± 1.9ADAS-cog, baseline 16.7 ± 2.3 20.6 ± 6.7ADAS-cog, week 4 15.2 ± 4.8 18.3 ± 4.5ADAS-cog, change −1.6 ± 4.4 −2.3 ± 3.4ADCS- ADL, baseline 63.5 ± 9.2 68.7 ± 6.8ADCS- ADL, week 4 67.7 ± 10.6 66.5 ± 6.7ADCS- ADL, change 4.2 ± 5.2 −2.2 ± 4.2CDR-SOB, baseline 5.6 ± 2.2 5.3 ± 1.9CDR-SOB, week 4 5.8 ± 2.4 5.0 ± 1.9CDR-SOB, change 0.2 ± 0.4 −0.3 ± 0.6NPI, baseline 9.5 ± 11.9 6.5 ± 6.1NPI, week 4 4.0 ± 5.0 7.7 ± 9.1NPI, change −5.5 ± 8.4 1.2 ± 5.1Values are shown as mean ± standard deviation (apart from sex). F and P values¶Kruskal-Wallis H test. ‡Analysis of completer subjects only. Week 4 ADCS-ADL an§Analysis of all randomized subjects, including AZD026 who discontinued study1.9, F = 2.07, P = 0.140; ADAS-cog change = 1.3 ± 3.5, F = 1.05, P = 0.393; NPI changAssessment Scale – cognitive subscale; ADCS-ADL, Alzheimer’s Disease CooperativRating Scale – Sum of Boxes; F, female; M, male; MMSE, Mini Mental Status Exami2 F, 4 M 3 F, 3 M14.3 ± 2.3 16.8 ± 3.4 0.84 † 0.4979.6 ± 17.9 65.4 ± 10.4 1.37 † 0.2822.2 ± 4.6 20.2 ± 2.3 0.63 † 0.6022.7 ± 4.0 22.5 ± 3.30.5 ± 3.4 2.3 ± 2.1 2.15 § 0.1323.8 ± 4.5 21.6 ± 7.1 1.73 † 0.1922.3 ± 8.1 23.0 ± 4.4−1.4 ± 5.4 1.4 ± 3.2 1.31 § 0.3067.2 ± 6.4 64.8 ± 3.1 0.67 † 0.5868.2 ± 7.6 66.4 ± 4.31.0 ± 2.0 1.6 ± 3.4 2.27 ‡ 0.124.8 ± 1.2 5.1 ± 1.8 0.22 † 0.884.8 ± 1.2 5.6 ± 1.90.0 ± 0.0 0.5 ± 0.4 2.60 ‡ 0.083.7 ± 4.3 13.2 ± 10.1 ¶ 0.372.5 ± 3.5 18.0 ± 8.9−1.2 ± 4.8 4.8 ± 7.6 ¶ 0.09for analysis of covariance, controlling for baseline score. †Analysis of variance.CDR-SOB invalid for subject AZD026, who was residing in skilled nursing facility.g after 9 days; excluding this subject, MMSE change in 125-mg group = 2.8 ±e = 1.8 ± 1.9, P = 0.169, Kruskal-Wallis H test. ADAS-cog, Alzheimer’s Diseasee Study – Activities of Daily Living Inventory; CDR-SOB, Clinical Dementianation; NPI, Neuropsychiatric Inventory.t gdogNygaard et al. Alzheimer's Research & Therapy  (2015) 7:35 Page 6 of 11Table 2 Number of participants in each AZD0530 treatmenTreatment group (AZD0539Adverse event Placebo 50 mAny adverse event* 3 4Diarrhea 2 0Headache 2 1Fatigue 1 0Nausea 2 0Pneumonia, atypical/bronchitis 0 0Congestive heart failure 0 0Renal insufficiency, worsened 0 0Elevated serum creatinine 0 0Cough, worsened 0 0Postnasal drip 0 0Pulmonary hypertension 0 0Anorexia 0 0Study medication was discontinued, and the subject wastreated with diuretics and antibiotics, gradually making afull recovery to baseline health. The overall conclusionwas that this event was possibly related to the study drug.Apart from this SAE, no other participant prematurelydiscontinued study medication.Two subjects receiving AZD0530 125 mg daily, one ofwhich experienced the SAE described, had significantlaboratory abnormalities that subsequently reversed:elevated serum creatinine, considered possibly relatedto study medication, and worsened renal insufficiency,unlikely related to study medication. There were nosignificant hematological changes, including platelet countsand absolute neutrophil counts. No significant changes inmeasures of vital signs or electrocardiogram readings wereseen across AZD0530 doses.AZD0530 drug levels in human cerebrospinal fluid atdifferent dosesWe compared the CSF drug levels across the AZD0530dosage groups. There was variability within any oneTinnitus 0 0Myalgias 0 0Squamous cell carcinoma 0 0Basal cell carcinoma 0 0Wrist pain 0 0Noncardiac chest pain 0 0Upper respiratory infection 0 1Flu-like symptoms 0 1Lightheadedness 0 1Vomiting 1 0*Includes participants who reported at least one adverse event. The effect of treatm(Fisher’s exact test, P > 0.05).roup experiencing an adverse eventse; n = 6 per group)100 mg 125 mg Total5 4 161 2 51 1 50 2 30 1 30 1 10 1 10 1 10 1 10 1 10 1 10 1 10 1 1group, but dose dependency was established (Figure 1A).The increase in CSF drug level was greater than linear,with a three-fold increase from 50 to 100 mg and a two-fold increase from 100 to 125 mg, most consistent withan exponential relationship. The trough CSF drug levelat 50 mg was 0.5 to 1.2 ng/ml (0.9 to 2.2 nM). Thetrough CSF drug level at 100 mg was 1.1 to 4.5 ng/ml(2.1 to 8.3 nM). The trough CSF drug level at 125 mgwas 1.4 to 7.6 ng/ml (2.5 to 14.0 nM). Using the ratio ofCSF:brain in mice measured to be 1:3, the estimatedhuman brain concentrations at 50 mg = 3 to 7 nM, at100 mg = 7 to 27 nM and at 125 mg = 8 to 46 nM. TheFyn Ki for AZD0530 is 5 to 10 nM. Analysis of the pre-clinical efficacy of AZD0530 in a mouse model of AD isreported elsewhere [43]; 5 mg/kg per day administeredfor 4 to 6 weeks reverses memory impairments in thismodel, corresponding to trough CSF levels in humansseen at doses of 100 to 125 mg/day (Figure 1A).We collected both CSF and plasma for AZD0530 druglevels at the week-4 endpoint. There was a close correl-ation between free AZD0530 drug levels in plasma and0 1 10 1 11 0 11 0 11 0 11 0 10 0 10 0 10 0 10 0 1ent group is not significant for any adverse event or individual adverse eventsMMSE was also administered at weekly visits as a safety125 mg daily (0.87 ± 1.86%) versus placebo (1.26 ± 2.24%;P = 0.71).Peripheral target engagementTo provide a readily accessible marker of peripheral Src0 10 20 30 40 5005101520Plasma AZD0530 (Free, nM)50mg100mg125mgBR2=0.97CSF AZD0530 (nM)AZD0530 Dose (mg)) Each point represents fasting trough cerebrospinal fluid (CSF) AZD0530trough CSF values are derived from brain levels at the 5 mg/kg per dayriance with Tukey post-hoc comparisons. (B) Relationship between CSF andifferent dose groups are illustrated with different colors. There is a tightrelation coefficient). Bars represent mean ± standard error of the mean.Nygaard et al. Alzheimer's Research & Therapy  (2015) 7:35 Page 7 of 11measure. No statistically significant effect of treatmentwas observed on any of clinical efficacy assessment mea-CSF AZD0530 (Figure 1B). CSF drug levels are aboutone-third of plasma free drug levels (Figure 1B).Clinical efficacy measuresAll subjects underwent standard clinical efficacy assess-ments at baseline and at the week-4 endpoint (Table 1).There was a tendency for MMSE scores to improve overtime, which may have been due to practice effects as theFigure 1 AZD0530 in human cerebrospinal fluid at different doses. (Alevel from a different human subject for the left three columns. The mousedose that rescued memory deficits [43]. **P < 0.001, one-way analysis of vaplasma AZD0530 levels. Each point is from a different individual, and dcorrelation of plasma and CSF level as indicated (R2 = 0.97, Pearson corn.s, Not significant.A50 100 125 Mouse051015CSF AZD0530 (nM)**** n.ssures, including ADAS-cog, ADCS-ADL, CDR-SOB, andNPI. The lack of a treatment effect was not surprisinggiven the small sample sizes and the short duration oftreatment, and with an investigational therapy that is beingdeveloped primarily for longer-term disease- modifyingpotential.18F-FDG PET measurementsThe effect of treatment with AZD0530 on 1-month re-ductions in CMRgl using statistical parametric mappingsROI was assessed using 18F-FDG PET imaging. Of the24 subjects, 22 underwent measurements of CMRgl atbaseline, and after 4 weeks on study drug or placebo.There was no effect of treatment group on change in aprespecified sROI consisting of voxels associated with pref-erential 12-month CMRgl declines in previously studiedpatients with the clinical diagnosis of mild-to-moderateAD dementia (F = 0.37, P = 0.78, df = 3.17, analysis of vari-ance). A post-hoc comparison of change in CMRgl for clin-ically relevant doses of AZD0530 (100 or 125 mg daily)versus placebo revealed no significant differences in therate of decline for CMRgl between AZD0530 100 tofamily kinase target inhibition, we assayed sCTX. This300n.s.****Placebo 50 100 1250100200AZD0530 dose (mg)% of baseline sCTXFigure 2 AZD0530 peripheral target engagement. Four-weektreatment with 100 mg and 125 mg AZD0530 reduces serum cross-linked C-telopeptide of type 1 collagen (sCTX), a marker of osteoclastactivity. Compared to placebo, mean difference in the change in sCTXfrom baseline for 50 mg was 51% (95% confidence interval, −22.0 to124.3), for 100 mg 85% (95% confidence interval, 8.2 to −161.6), and for125 mg 83% (95% confidence interval, 6.2 to 159.6). **P< 0.05, analysisof variance with Dunnet’s multiple comparisons test. n.s., Not significant.collagen fragment is a marker of activity for the osteoclast,a cell in which Src plays an important regulatory role.AZD0530 is reported to decrease sCTX in humans in adose-dependent fashion [38]. Our results confirm Src per-ipheral target engagement, with a decrease in serum sCTXafter 4 weeks of daily 100- to 125-mg AZD0530 adminis-tration (Figure 2).AZD0530 does not inhibit Abl kinase at doses up to 125 mgAZD0530 has highest activity against Src family kinases,with little or no potency against a panel of other kinases.The only other detectable activity is against Abl kinase,with a Ki approximately 15-fold higher [28,44]. Therefore,we measured the activity of plasma inhibitory activity ofAZD0530 against Abl kinase, by monitoring the level ofStat5 phosphorylation as a substrate of Abl in leukemiacells as described [40,41]. While the Abl kinase inhibi-tor STI-571 inhibits Abl to Stat5 signaling with a halfmaximal inhibitory concentration (IC50) of 1.6 nM incells incubated with plasma, the IC50 for AZD0530 isnearly 100-fold higher, at 156 nM (Figure 3A,B). Plasmasamples from subjects treated with 0, 50, 100 and 125 mgwere incubated with the leukemia cells in vitro, andpStat5 measured as a read out of Abl kinase activity(Figure 3C,D). No significant inhibition of Abl kinasewas observed. Thus, at these doses of AZD0530, kinase in-hibition is specific for the Src family (Figure 2), withoutsignificant alterations in Abl kinase activity (Figure 3D).Cerebrospinal fluid biomarkersLevels of total Aβ40, Aβ42, Tau, and p-Tau were assessedin CSF samples from all subjects in the study after 4 weeksof study medication. Although we did not obtain CSF atbaseline to analyze drug effects on CSF biomarkers, wefound no difference in levels of any AD biomarker atthe 4-week endpoint between subjects treated with 100to 125 mg AZD0530 and placebo (for Aβ40: placebo,4,876 pg/ml (±789 pg/ml); AZD0530 100 mg, 3,963 pg/ml(±624 pg/ml); AZD0530 125 mg, 4,501 pg/ml (±311 pg/ml); for Aβ42: placebo, 601 pg/ml (±81 pg/ml); AZD0530100 mg, 523 pg/ml (±48 pg/ml); AZD0530 125 mg,553 pg/ml (±34 pg/ml); for total tau: placebo, 0.53 ng/ml(±0.17 ng/ml); AZD0530 100 mg, 0.57 ng/ml (±0.20 ng/ml); AZD0530 125 mg, 0.57 ng/ml (±0.14 ng/ml); for p(T231)-Tau: placebo, 5.33 nM (±2.29 nM); AZD0530100 mg, 5.88 nM (±1.89 nM); AZD0530 125 mg, 5.21 nM(±1.80 nM)).blisetesuNygaard et al. Alzheimer's Research & Therapy  (2015) 7:35 Page 8 of 11Figure 3 Plasma from AZD0530-treated subjects does not inhibit Acells incubated with the indicated concentrations of AZD0530 or STI-571microdensitometry and a half maximal inhibitory concentration (IC50) dwas detected by pStat5 level in K-562 cells incubated with plasma fromAbl inhibitory activity determined by pStat5 level was measured in all subjectdifference between groups was observed (n.s.).signaling to Stat5. (A) The level of pStat5 and total Stat5 in K-562shown. (B) The pStat5 level from experiments in (A) was measured byrmined for each compound. (C) The plasma inhibitory activity for Ablbjects treated with the indicated doses of AZD0530. (D) The plasmas, and is plotted as mean ± standard error of the mean. No significantNygaard et al. Alzheimer's Research & Therapy  (2015) 7:35 Page 9 of 11DiscussionPreclinical studies in both rodent models and patientswith AD indicate that Fyn may be a promising target fornovel therapeutic intervention [23]. Here, we report aPhase Ib study to explore the use of a Fyn-specific Srcfamily kinase inhibitor for the treatment of patients withAD. Our study shows that 100 mg and 125 mg dosesachieved CSF levels corresponding to brain levels thatrescued memory deficits in transgenic AD mouse models.A therapeutic benefit was not expected after a 1-monthtreatment period, and no treatment effect was observedacross doses of AZD0530 on measures of cognitive andneuropsychiatric function, activities of daily living, or cere-bral glucose metabolism.Some safety and tolerability issues may emerge, particu-larly at the 125 mg dose. However, the tight correlationbetween plasma and CSF levels of AZD0530 will enableindividualized dosing within the 100 to 125 mg range,based on early plasma level monitoring. Individualizeddosing may maximize the number of subjects whoreach the target CSF drug concentration of 5 nM, whileminimizing those who encounter safety and tolerabilityproblems. Fyn regulates a diverse set of cellular func-tions, including cell proliferation, migration, and differ-entiation, synaptic function, CNS myelination, T cellsignaling, and platelet function [23], emphasizing notonly the importance of careful safety and tolerabilitymonitoring in larger clinical trials, but also the goal ofnormalizing aberrant Fyn activity in AD, without signifi-cantly altering its physiologic functions. Previous experi-ence with AZD0530 has indicated a possible, but rare,relationship with interstitial lung disease in patients withadvanced solid tumors [33]. The single SAE in this studyinvolved a case of congestive heart failure and pneumonia,for which a drug-induced pneumonitis could not be en-tirely ruled out. Although no conclusive link has beenestablished between AZD0530 and drug-induced pneu-monitis, future trials of AZD0530 for AD should continueto monitor this possibility with thoracic high-resolutioncomputed tomography for any unexplained pulmonarysymptoms. We recognize the limitations of our results dueto the relatively small sample size and the short duration oftreatment. These limitations will be addressed by a largerPhase IIa clinical trial of AZD0530 in AD (NCT02167256).Masitinib, a tyrosine kinase inhibitor selective for c-Kit,platelet-derived growth factor, Lyn, and to a lesser degreeFyn, was recently used in a 24-week, Phase II dose-rangingtrial in France, involving 34 patients with mild-to-moderateAD [45]. The trial showed reasonable tolerability, and drugtreatment was associated with improvements in cognitionand daily function at 12 and 24 weeks. A large internationalPhase III trial was launched in 2013 to evaluate the efficacyand safety of two doses of masitinib compared to placebo(NCT01872598). The Phase II data provide further clinicalsupport for tyrosine kinase inhibition as a treatment strat-egy in AD.ConclusionsThe current Phase Ib trial demonstrates that AZD0530is generally safe and well tolerated in patients with AD,with oral dosing yielding excellent CNS drug concentra-tion. Together with our preclinical data, the Phase Ibstudy results support the dosing regimen for a larger on-going Phase IIa trial in AD of 100 to 125 mg AZD0530daily (NCT02167256).AbbreviationsAβ: amyloid-beta; Aβo: oligomeric amyloid-beta; AD: Alzheimer’s disease;ADAS-cog: Alzheimer’s Disease Assessment Scale – cognitive subscale;ADCS-ADL: Alzheimer’s Disease Cooperative Study – Activities of DailyLiving Inventory; CDR-SOB: Clinical Dementia Rating Scale – Sum of Boxes;CMRgl: cerebral metabolic rate for glucose; CNS: central nervous system;CSF: cerebrospinal fluid; FDG: fluorodeoxyglucose; IC50: half maximal inhibitoryconcentration; MMSE: Mini-Mental State Examination; NPI: NeuropsychiatricInventory; PET: positron emission tomography; PrPC: cellular prion protein;SAE: serious adverse event; sCTX: serum C-telopeptide of type 1 collagen;sROI: statistical region of interest; ULN: upper limit of normal.Competing interestsSMS is a co-founder of Axerion Therapeutics, seeking to develop NgR- andPrP-based therapeutics. In relation to Alzheimer’s disease therapeutics, CHvDhas served as a scientific advisor or consultant to Bristol-Myers Squibb, JanssenAlzheimer Immunotherapy, Pfizer, GlaxoSmithKline, Elan Pharmaceuticals, RochePharmaceuticals, and Abbott/AbbVie Inc. CHvD has received researchsupport from Bristol-Myers Squibb, Elan Pharmaceuticals, Janssen AlzheimerImmunotherapy, Pfizer, Eli Lilly, Merck, Baxter Pharmaceuticals, GlaxoSmithKline,Abbott Laboratories, Medivation, Biogen Idec, Eisai, Genentech, and RochePharmaceuticals. EMR has been a scientific advisor for Alzheon, AstraZeneca,CereSpir, Eisai, Eli Lilly, GlaxoSmithKline, and Sanofi. EMR receives researchsupport from Avid Radiopharmaceuticals/Eli Lilly, Genentech Inc., and Novartis.The other authors declare that they have no competing interests.Authors’ contributionsHBN, SMS and CHvD conceived of the study. HBN, AFW, GSB, SPG, MGM,SMS and CHvD participated in the overall design, execution, and analysis ofthe study. KAS performed and analyzed AD biomarker analysis. ACK performedand analyzed rodent pharmacokinetic studies. BJR performed the Abl kinaseassay and statistical analysis. TSK and HBN performed and analyzed sCTXenzyme-linked immunosorbent assays. PV performed and analyzed neuroimaging.AJK supervised and analyzed the Abl kinase assay and statistical analysis. KC andEMR designed the PET imaging protocol and statistical analysis. HBN, SMS, andCHvD drafted the manuscript. All authors reviewed the manuscript, providedcritical input, and approved the final manuscript.AcknowledgementsWe thank AstraZeneca for providing AZD0530 compound and for helpfuladvice. We acknowledge support from the National Institutes of Health toHBN, CHvD and SMS, and from the Falk Medical Research Trust to SMS.Author details1Alzheimer’s Disease Research Unit, Yale University School of Medicine, NewHaven, Connecticut, USA. 2Department of Neurology, Yale University Schoolof Medicine, New Haven, Connecticut, USA. 3Program in CellularNeuroscience, Neurodegeneration and Repair (CNNR), Yale University Schoolof Medicine, New Haven, Connecticut, USA. 4Department of Psychiatry, YaleUniversity School of Medicine, New Haven, Connecticut, USA. 5Departmentof Molecular Biophysics and Biochemistry, Yale University School of Medicine,New Haven, Connecticut, USA. 6Department of Diagnostic Radiology, YaleUniversity School of Medicine, New Haven, Connecticut, USA. 7Banner8Alzheimer’s Institute, Phoenix, Arizona, USA. Current address: University ofBritish Columbia, Division of Neurology, Djavad Mowafaghian Centre forBrain Health, Vancouver, Canada.Nygaard et al. Alzheimer's Research & Therapy  (2015) 7:35 Page 10 of 11Received: 6 November 2014 Accepted: 16 March 2015References1. Nygaard HB. Current and emerging therapies for Alzheimer’s disease. ClinTher. 2013;35:1480–9.2. Puzzo D, Privitera L, Leznik E, Fa M, Staniszewski A, Palmeri A, et al. Picomolaramyloid-beta positively modulates synaptic plasticity and memory inhippocampus. J Neurosci. 2008;28:14537–45.3. Giuffrida ML, Caraci F, Pignataro B, Cataldo S, De Bona P, Bruno V,et al. Beta-amyloid monomers are neuroprotective. J Neurosci.2009;29:10582–7.4. Walsh DM, Klyubin I, Fadeeva JV, Cullen WK, Anwyl R, Wolfe MS, et al.Naturally secreted oligomers of amyloid beta protein potently inhibithippocampal long-term potentiation in vivo. Nature. 2002;416:535–9.5. Lauren J, Gimbel DA, Nygaard HB, Gilbert JW, Strittmatter SM. Cellular prionprotein mediates impairment of synaptic plasticity by amyloid-beta oligomers.Nature. 2009;457:1128–32.6. Lacor PN, Buniel MC, Furlow PW, Clemente AS, Velasco PT, Wood M, et al.Abeta oligomer-induced aberrations in synapse composition, shape, anddensity provide a molecular basis for loss of connectivity in Alzheimer’sdisease. J Neurosci. 2007;27:796–807.7. Chen S, Yadav SP, Surewicz WK. Interaction between human prion proteinand amyloid-beta (Abeta) oligomers: role of N-terminal residues. J BiolChem. 2010;285:26377–83.8. Calella AM, Farinelli M, Nuvolone M, Mirante O, Moos R, Falsig J, et al. Prionprotein and Abeta-related synaptic toxicity impairment. EMBO Mol Med.2010;2:306–14.9. Balducci C, Beeg M, Stravalaci M, Bastone A, Sclip A, Biasini E, et al. Syntheticamyloid-beta oligomers impair long-term memory independently of cellularprion protein. Proc Natl Acad Sci U S A. 2010;107:2295–300.10. Um JW, Nygaard HB, Heiss JK, Kostylev MA, Stagi M, Vortmeyer A, et al.Alzheimer amyloid-beta oligomer bound to postsynaptic prion proteinactivates Fyn to impair neurons. Nat Neurosci. 2012;15:1227–35.11. Resenberger UK, Harmeier A, Woerner AC, Goodman JL, Muller V, Krishnan R,et al. The cellular prion protein mediates neurotoxic signalling of beta-sheet-richconformers independent of prion replication. EMBO J. 2011;30:2057–70.12. Bate C, Williams A. Amyloid-beta-induced synapse damage is mediated viacross-linkage of cellular prion proteins. J Biol Chem. 2011;286:37955–63.13. Alier K, Ma L, Yang J, Westaway D, Jhamandas JH. Abeta inhibition of ionicconductance in mouse basal forebrain neurons is dependent upon thecellular prion protein PrPC. J Neurosci. 2011;31:16292–7.14. Chung E, Ji Y, Sun Y, Kascsak RJ, Kascsak RB, Mehta PD, et al. Anti-PrPCmonoclonal antibody infusion as a novel treatment for cognitive deficits inan Alzheimer’s disease model mouse. BMC Neurosci. 2010;11:130.15. Gimbel DA, Nygaard HB, Coffey EE, Gunther EC, Lauren J, Gimbel ZA, et al.Memory impairment in transgenic Alzheimer mice requires cellular prionprotein. J Neurosci. 2010;30:6367–74.16. Kudo W, Lee HP, Zou WQ, Wang X, Perry G, Zhu X, et al. Cellular prionprotein is essential for oligomeric amyloid-beta-induced neuronal cell death.Hum Mol Genet. 2012;21:1138–44.17. You H, Tsutsui S, Hameed S, Kannanayakal TJ, Chen L, Xia P, et al. Abetaneurotoxicity depends on interactions between copper ions, prionprotein, and N-methyl-D-aspartate receptors. Proc Natl Acad Sci U S A.2012;109:1737–42.18. Zou WQ, Xiao X, Yuan J, Puoti G, Fujioka H, Wang X, et al. Amyloid-beta42interacts mainly with insoluble prion protein in the Alzheimer brain. J BiolChem. 2011;286:15095–105.19. Freir DB, Nicoll AJ, Klyubin I, Panico S, McDonald JM, Risse E, et al. Interactionbetween prion protein and toxic amyloid beta assemblies can betherapeutically targeted at multiple sites. Nat Commun. 2011;2:336.20. Barry AE, Klyubin I, Mc Donald JM, Mably AJ, Farrell MA, Scott M, et al.Alzheimer’s disease brain-derived amyloid-beta-mediated inhibition of LTPin vivo is prevented by immunotargeting cellular prion protein. J Neurosci.2011;31:7259–63.21. Um JW, Kaufman AC, Kostylev M, Heiss JK, Stagi M, Takahashi H, et al.Metabotropic glutamate receptor 5 is a coreceptor for Alzheimer abetaoligomer bound to cellular prion protein. Neuron. 2013;79:887–902.22. Larson M, Sherman MA, Amar F, Nuvolone M, Schneider JA, Bennett DA, et al.The complex PrP(c)-Fyn couples human oligomeric Abeta with pathologicaltau changes in Alzheimer’s disease. J Neurosci. 2012;32:16857–16871a.23. Nygaard HB, van Dyck CH, Strittmatter SM. Fyn kinase inhibition as a noveltherapy for Alzheimer’s disease. Alzheimers Res Ther. 2014;6:8.24. Bhaskar K, Hobbs GA, Yen SH, Lee G. Tyrosine phosphorylation of tauaccompanies disease progression in transgenic mouse models oftauopathy. Neuropathol Appl Neurobiol. 2010;36:462–77.25. Bhaskar K, Yen SH, Lee G. Disease-related modifications in tau affect theinteraction between Fyn and Tau. J Biol Chem. 2005;280:35119–25.26. Lee G, Thangavel R, Sharma VM, Litersky JM, Bhaskar K, Fang SM, et al.Phosphorylation of tau by fyn: implications for Alzheimer’s disease. JNeurosci. 2004;24:2304–12.27. Lee G, Newman ST, Gard DL, Band H, Panchamoorthy G. Tau interacts withsrc-family non-receptor tyrosine kinases. J Cell Sci. 1998;111:3167–77.28. Hennequin LF, Allen J, Breed J, Curwen J, Fennell M, Green TP, et al.N-(5-chloro-1,3-benzodioxol-4-yl)-7-[2-(4-methylpiperazin-1-yl)ethoxy]-5-(tetrahydro-2H-pyran-4-yloxy)quinazolin-4-amine, a novel, highly selective,orally available, dual-specific c-Src/Abl kinase inhibitor. J Med Chem.2006;49:6465–88.29. McKhann GM, Knopman DS, Chertkow H, Hyman BT, Jack Jr CR, Kawas CH,et al. The diagnosis of dementia due to Alzheimer’s disease: recommendationsfrom the National Institute on Aging-Alzheimer’s Association workgroupson diagnostic guidelines for Alzheimer’s disease. Alzheimers Dement.2011;7:263–9.30. Folstein MF, Folstein SE, McHugh PR. “Mini-mental state”. A practical methodfor grading the cognitive state of patients for the clinician. J Psychiatric Res.1975;12:189–98.31. Sheikh J, Yesavage J. Geriatric Depression Scale (GDS): recent evidence anddevelopment of a shorter version. In: Brind TL, editor. Clinical gerontology:a guide to assessment and intervention. New York: Haworth Press; 1986.p. 165–73.32. Rosen WG, Terry RD, Fuld PA, Katzman R, Peck A. Pathologicalverification of ischemic score in differentiation of dementias. AnnNeurol. 1980;7:486–8.33. Baselga J, Cervantes A, Martinelli E, Chirivella I, Hoekman K, Hurwitz HI, et al.Phase I safety, pharmacokinetics, and inhibition of SRC activity studyof saracatinib in patients with solid tumors. Clin Cancer Res.2010;16:4876–83.34. Rosen WG, Mohs RC, Davis KL. A new rating scale for Alzheimer’s disease.Am J Psychiatry. 1984;141:1356–64.35. Galasko D, Bennett D, Sano M, Ernesto C, Thomas R, Grundman M, et al. Aninventory to assess activities of daily living for clinical trials in Alzheimer’sdisease. The Alzheimer’s Disease Cooperative Study. Alzheimer Dis AssocDisord. 1997;11:S33–9.36. Cummings JL. The Neuropsychiatric Inventory: assessing psychopathologyin dementia patients. Neurology. 1997;48:S10–6.37. Morris JC. The Clinical Dementia Rating (CDR): current version and scoringrules. Neurology. 1993;43:2412–4.38. Hannon RA, Clack G, Rimmer M, Swaisland A, Lockton JA, Finkelman RD,et al. Effects of the Src kinase inhibitor saracatinib (AZD0530) on boneturnover in healthy men: a randomized, double-blind, placebo-controlled, multiple-ascending-dose phase I trial. J Bone Miner Res.2010;25:463–71.39. Hannon RA, Finkelman RD, Clack G, Iacona RB, Rimmer M, Gossiel F, et al.Effects of Src kinase inhibition by saracatinib (AZD0530) on bone turnoverin advanced malignancy in a Phase I study. Bone. 2012;50:885–92.40. Ishikawa Y, Kiyoi H, Watanabe K, Miyamura K, Nakano Y, Kitamura K,et al. Trough plasma concentration of imatinib reflects BCR-ABL kinaseinhibitory activity and clinical response in chronic-phase chronicmyeloid leukemia: a report from the BINGO study. Cancer Sci.2010;101:2186–92.41. Podesta JE, Sugar R, Squires M, Linardopoulos S, Pearson AD, Moore AS.Adaptation of the plasma inhibitory activity assay to detect Aurora, ABL andFLT3 kinase inhibition by AT9283 in pediatric leukemia. Leuk Res.2011;35:1273–5.42. Chen K, Langbaum JB, Fleisher AS, Ayutyanont N, Reschke C, Lee W, et al.Twelve-month metabolic declines in probable Alzheimer’s disease andamnestic mild cognitive impairment assessed using an empirically pre-defined statistical region-of-interest: findings from the Alzheimer’s DiseaseNeuroimaging Initiative. Neuroimage. 2010;51:654–64.43. Kaufman AC, Salazar SV, Haas LT, Yang J, Kostylev MA, Jeng AT, et al. Fyninhibition rescues established memory and synapse loss in Alzheimer mice.Ann Neurol. 2015; doi:10.1002/ana.24394.44. Green TP, Fennell M, Whittaker R, Curwen J, Jacobs V, Allen J, et al.Preclinical anticancer activity of the potent, oral Src inhibitor AZD0530.Mol Oncol. 2009;3:248–61.45. Piette F, Belmin J, Vincent H, Schmidt N, Pariel S, Verny M, et al. Masitinib asan adjunct therapy for mild-to-moderate Alzheimer’s disease: a randomised,placebo-controlled phase 2 trial. Alzheimers Res Ther. 2011;3:16.Submit your next manuscript to BioMed Centraland take full advantage of: • Convenient online submission• Thorough peer review• No space constraints or color figure charges• Immediate publication on acceptance• Inclusion in PubMed, CAS, Scopus and Google Scholar• Research which is freely available for redistributionNygaard et al. Alzheimer's Research & Therapy  (2015) 7:35 Page 11 of 11Submit your manuscript at www.biomedcentral.com/submit


Citation Scheme:


Citations by CSL (citeproc-js)

Usage Statistics



Customize your widget with the following options, then copy and paste the code below into the HTML of your page to embed this item in your website.
                            <div id="ubcOpenCollectionsWidgetDisplay">
                            <script id="ubcOpenCollectionsWidget"
                            async >
IIIF logo Our image viewer uses the IIIF 2.0 standard. To load this item in other compatible viewers, use this url:


Related Items